FESBE Apresentados - Sexta

XXVI Reunio Anual da FeSBE - FeSBE 2011
Resumo:02-077
AEROBIC EXERCISE TRAINING INDUCED CARDIAC HYPERTROPHY INVOLVES REGULATORY
MICRORNAS, DECREASED ACE-ANG II, AND SYNERGISTIC REGULATION OF ACE2-ANG (1-7)
Fernandes, T. 1; Hashimoto, N. Y. 1; Magalhes, F. C. 1; Fernandes, F. B. 3; Casarini, D. E. 3; Carmona, A.

3
; Krieger, J. E. 2; Phillips, M. I. 4; Oliveira, E. M. 1
1
Laboratory of Biochemistry of the Motor Activity - USP, EEFE-USP
2
Laboratory of Genetics and Molecular Cardiology - InCor , FMUSP
3
Nephrology Division, Kidney and Hypertension Hospital , UNIFESP
4
Laboratory of Stem Cells / Keck Graduate Institute, KGI
Objectives:
Aerobic exercise training (ET) leads to a physiological, non pathological left ventricular hypertrophy (LVH); however, the
underlying biochemical and molecular mechanisms of physiological LVH are unknown. MicroRNAs may be important for LVH,
since they function as negative regulators of gene expression by inhibiting translation or promoting degradation of target mRNAs.
The role of microRNAs regulating the classic and the novel cardiac renin angiotensin system (RAS) was studied in exerciseinduced LVH.
Methods and Results:
Wistar rats (n=30) were assigned to three groups: sedentary (S), Trained 1 (T1) and Trained 2. T1: swimming training consisted
of 60 min 1x/day/10 weeks, with 5% body weight overload. T2 the same as T1 until 8th week, in the 9th week they trained
2x/day and in the 10th week 3x/day. RAS involvement in the left LVH induced by ET was analyzed using Angiotensin II type 1
receptor (AT1) blockade (Losartan- 20 mg/kg/day) during ET protocol. Blood pressure (BP) and heart rate (HR) were evaluated
by direct measurement, LVH by tissue weight/body weight ratio, myocyte diameter by histological method, angiotensinconverting enzyme 1 (ACE) and 2 (ACE2) activity, gene and protein expression by fluorometric method, real-time PCR and
western blot, respectively. Plasma renin activity (PRA) by radioimmunoassay and angiotensin I, II and (1-7) by HPLC. AT1 and
AT2 receptors gene and protein expression also were studied. MicroRNAs profile was analyzed by microarray and it that targeted
RAS genes were validated by real-time PCR. Results were presented as meanSEM. BP was unchanged while resting HR
decreased in all trained groups. LVH obtained by T1 and T2 training protocol was 17% (p
Conclusions:
LVH induced by ET involves microRNAs regulation and an increase in cardiac AT1 receptor without the participation of Ang II.
Parallel to this, increase in ACE2, Ang (1-7) and AT2 receptor in the heart by exercise suggests that this non classic cardiac RAS
counteracts the classic cardiac RAS activating local vasodilatation pathway. Together these effects might provide the additional
aerobic capacity required by the exercised heart.
Keywords: aerobic exercise training, AT1 receptor blockade, cardiac hypertrophy, microRNAs, renin angiotensin system
Financial Support: FAPESP, CAPES and CNPq
Resumo:02-078
THE ROLE OF TYPE 2 ANGIOTENSIN II RECEPTOR (AT2) IN THE CARDIOPROTECTIVE RESPONSE

INDUCED BY THYROID HORMONE.
Tavares, F. M. ; Barreto-chaves, M. L. M
Department of Anatomy, Institute of Biomedical Sciences, USP
Objectives:
To evaluate the role of AT2 receptor in the recovery of cardiac post-ischemic function in a hyperthyroidism model.
Thriodothyronine (T3) (0.7 g/Kg BW/day, i.p.) was injected to male adult Wistar rats in the presence or absence of AT2R
blocker (PD123319) for 14 days. AT2R blocker (10 mg/Kg/day) was administered subcutaneously through the implantation of
osmotic minipumps (Alzet). Animals were subdivided in four different groups: 1) Control: rats treated with normal saline (n=9);
2) PD (n=6); 3) T3 (n=7) and 4) PD plus T3 (n=4). Animals from each group were used for the assessment of baseline cardiac
performance and to response to ischemia-reperfusion (I-R) protocols. Hearts were rapidly excised and mounted on a Langendorff
apparatus. Isolated hearts were perfused in a retrograde way at a constant flow with oxygenated (95% O2, 5% CO2)
KrebsHenseleit buffer at a temperature of 37C. A waterfilled balloon was inserted in the left ventricular cavity to assess the
following parameters: left ventricular pressure, dP/dT max e min, heart rate and perfusion pressure. After 30 min of stabilization,
hearts were subjected to 20 min of zeroflow global ischemia and 45 min of reperfusion. Cardiac hypertrophy was determined
based on heart weight/body weight and heart weight/tibia length ratios. The results demonstrate that T3 induced cardiac
hypertrophy (25% vs control p
Conclusions:
In conclusion, we observed that the blockade of AT2R with PD 123319 induced cardioprotective response, however, it abolished
the cardioprotection induced by T3.
Keywords: AT2 receptor, Cardioprotection, Hyperthyroidism, Ischemia/Reperfusion injury
Financial Support: FAPESP, CNPq.
Resumo:02-079
PLATELET AGGREGATION IS REDUCED DURING EXPERIMENTAL SEPSIS
Gonalves, M. C. ; Sordi, R. ; Assreuy, J.

Department of Pharmacology / Universidade Federal de Santa C, UFSC
Objectives:
Sepsis is a systemic inflammatory state associated with an infection caused by bacteria, fungi, viruses or parasites. Many
mechanisms are involved at sepsis physiopathology including cytokine release, activation of neutrophils, monocytes, endothelial
cells and also activation of coagulation and fibrinolysis systems. Imbalance between procoagulant and anticoagulant processes
during sepsis has been described. This imbalance is likely to be an important event contributing to multiple organ dysfunctions.
Although there are some reports showing that in human sepsis a reduction in platelet numbers occur, the role of platelets in sepsis
remains unknown. In this study we have attempted to characterize the platelet aggregation in experimental sepsis induced by
cecal ligation and puncture (CLP) in male rats temporally.
All procedures have been approved by the institutional animal ethics committee (CEUA/UFSC, Protocol PP00595). About 5 mL
of blood were collected from anaesthetized septic and nave rats via cardiac puncture with sodium citrate 3.8% (1:9 ratio,
citrate:blood) as anticoagulant. Platelet rich plasma (PRP) was obtained by centrifugation and aggregation assay was performed
in an Infinite Plate Reader (Tecan, Switzerland). About 50.10 6 platelets per well (96-well plate) were assayed during 15 minutes,
at 37C under moderate and constant shaking. The platelet aggregation was induced by adding 5 M of adenosine diphosphate
(ADP) or collagen (130 g/mL) in PRP from nave and septic animals (6 h, 12 h and 24 h after CLP surgery). The platelet
aggregation was quantified by optical density (OD) at 650 nm. The difference between the basal and peak aggregation OD was
expressed in milliunits of OD (mUOD). The platelet aggregation induced by ADP in septic rats was diminished in all CLP
groups, and it was statistically significant at CLP 12 h group (mUOD: 77.7 0.02; n=6) when compared to nave group (127.1
0.01; n= 6). Aggregation induced by collagen also decreased substantially 12 h (85.6 0.02; n=6) and 24 h after CLP surgery
(71.1 0.004; n=6) when compared to the nave group (180.7 0.02; n=6). Nitrate + nitrite (NOX) levels were increased in
plasma samples of septic animals (76.8 14.1, 135.6 18.9 and 90.2 27.4 M, 6 h, 12 h and 24 h, respectively) when
compared to nave values (34.8 25).
Conclusions:
Septic animal exhibited lower platelet aggregation when compared to nave rats. The reduced platelet aggregation ability might
be caused by the large amount of nitric oxide (NO) produced in sepsis. This platelet dysfunction may be contributing to the
hemostatic imbalance of sepsis.
Keywords: Hemostasis, Nitric oxide, Platelet aggregation, Sepsis
Financial Support: CNPq, CAPES and FAPESC.
Resumo:02-080
CHRONIC MALNUTRITION PROMOTED BY THE BRAZILIAN NORTHEAST DIET ALTERS CARDIAC
MORPHOLOGY AND FUNCTION IN RATS.
Mendes, L. V. P. 1; Dias,m. S. 1; Boldrini, L. C1; Takiya, C. M. 1; Oliveira-pinto, L. M 1; Nascimento, J. H.

M. 1; Lamas, M. E. 1; Vieyra, A. 1; Cunha, V. M. N. 1; Lara, L. S1
1
Instituto de Cincias Biomdicas, UFRJ
2
Instituto de Biofsica Carlos Chagas Filho, UFRJ
3
Programa de Ps-Graduao em Cincias Morfolgicas, UFRJ
Objectives:
In previous work, we showed that malnutrition promoted for regional basic diet (RBD) from Brazilian northeast in chronic period
of development (RBD-C) modifies the homeostasis of intracellular Ca2+. Such changes suggest the development of a heart
failure process in the RBD-C group. The aims of the present work were: 1- evaluate the contractility of the hearts from RDB-C
rats; 2- assess histologically the heart tissue from RBD-C rats; 3- evaluate the content of free sulfhydryl essential for the catalytic
activity of Ca2+-ATPases

Male Wistar rats from healthy mothers after weaning were subjected to the diet for 10 weeks RDB (RDB-C, n = 9), while the rats
from control group were fed a conventional diet (Cont, n = 9). After a period of 13 weeks of age the animals were sacrificed
(CEUA DFBCICB007). After the sacrifice, the heart was used in Langendorff assays, using increasing doses of Isoproterenol.
The values of LVP from hearts of RBD-C group were not different from controls (for 1 M isoproterenol, 135.4 26.0 vs 133.7
25.5 %, respectively, n=5 and n=3, P>0.05). The contractility index calculated for maximum rate of contraction (+dP/dTmax)
was increased in the group RBD-C compared with control group (for 100 nM Isoproterenol, 217.884 vs 68.614.9, respectively,
P< 0.05). Histological analysis was performed according to Masson's trichrome method, in which atrophy of cardiomiocytes,
inflammatory infiltration, and fat cells were observed in the hearts of rats from RDB-C group. The Ca2+-ATPase activity was
also measured in heart homogenates using gama [32P-ATP]. It was observed a significant increase of the total Ca2+-stimulated
activity in RDB-C group in relation to the control group (2136 25.4 vs 985.6 30 nmolPi / mg.h-1, respectively, n=3, P
Conclusions:
The increase of the contractility index in animals RBD-C indicates an attempt to improve the working heart since the cardiac
output is reduced in this group (reported earlier). Changes in thickness of cardiomiocytes as well as the presence of inflammatory
infiltration indicate the installation of necrosis process in the hearts from RBD-C rats. As reported before using microsomes, the
increased of the total Ca2+-ATPase activity of heart homogenates indicates that somehow the heart cells from RDB-C rats try to
maintain the cardiac contractility and the efficient remove of Ca2+ ions. The loss of reduced sulfhydryls indicates enzymatic
oxidation associated to the loss of catalytic activity. In relation to Ca2+-ATPases, it is possible that the decrease of free
sulfhydryls is associated with SERCA oxidation because it was observed previously a significant reduction of the activity of this
enzyme without any change of protein expression.
Keywords: atrophy, heart, langendorff, malnutrition, Regional Basic Diet
Financial Support: Projeto Casadinho-CNPq; PROCAD-CAPES; FAPERJ Primeiros Projetos, Programa

ALV.
Resumo:02-081
TIME COURSE AND EFFICACY OF MK-467 BLOCKING THE HEMODYNAMIC RESPONSES CAUSED BY THE
SELECTIVE 2 ADRENERGIC AGONIST CLONIDINE.
Dias, L. O. 2,1; Durand, M. T. 1; Castania, J. A. 1; Fazan Jr. , R. 1; Salgado, H. C. 1

1
Department of Physiology, FMRP/USP
2
Universidade Estadual de Santa Cruz, UESC
Objectives:
MK-467, a benzofuroquinolizine derivative, has been considered a selective 2 adrenergic receptor antagonist that can be useful
to examine the role played by 2 adrenergic receptors along with the 1 counterparts in the control of arterial pressure (AP). In
addition, it has not been described, precisely, how long MK-467 blocks 2 adrenergic receptors.In the present study it was
evaluated, in conscious freely moving rats, the time course and the efficacy of MK-467 to block the hypertensive response and
the reflex bradycardia caused by the selective 2 adrenergic agonist clonidine. In addition, it was also investigated a possible
effect of MK-467 on 1 adrenergic receptors by the exam of the hypertensive response and the reflex bradycardia elicited by the
selective 1 adrenergic agonist phenylephrine.
Wistar rats (200-300g) were anesthetized (tribromoethanol 250 mg/kg, ip) and the femoral artery and vein were catheterized for
AP recording and drug administration, respectively. Twenty-four hours after the surgical procedure, the experiments were
conducted in conscious freely moving rats. The protocol consisted of basal AP and heart rate (HR) recordings followed by
phenylephrine (4 g/kg) and clonidine (1.5 mg/kg) injections, before and after MK-467 administration (3 mg/kg). After MK-467
administration clonidine was injected at 5, 15, 30, 45 and 60 minutes while phenylephrine was injected only at 35 minutes. MK467 produced a significant fall in basal AP, which occurred in the first 5 minutes (Basal: 103 4 vs. MK-467: 93 4 mmHg)
eliciting a reflex tachycardia (Basal: 353 13 vs. MK-467: 451 26 bpm). Nevertheless, these responses were not maintained,
and AP returned to basal values within 15 minutes. The hypertensive response (MAP: 40 3 mmHg) and the reflex bradycardia
(HR: -58 9 bpm) to clonidine were blocked after MK-467 at 5 min (1 1 mmHg and -2 5 bpm), 15 min ( 3 1 mmHg and
+1 7 bpm), 30 min (7 3 mmHg and -6 9 bpm), 45 min ( 8 4 mmHg and -1 6 bpm) and 60 min (5 4 mmHg and -10 5
bpm). On the other hand, MK-467 did not affect the hypertensive response (44 3 vs. 35 5 mmHg after) and the reflex
bradycardia (-58 11 vs. -33 10 bpm after) to phenylephrine.
Conclusions:
The results demonstrate that MK-467 produced a prompt fall in AP combined with reflex tachycardia. In addition, MK-467
promoted peripheral 2 adrenergic receptor blockade which persisted for 60 minutes. Moreover, the administration of MK-467
did not block the response to 1 adrenergic receptor activation with phenylephrine, showing remarkable selectivity to peripheral
2 adrenergic receptor.
Keywords: 2 adrenergic receptor, MK-467, Clonidine
Financial Support: Capes, CNPq, FAPESP
Resumo:02-082
THERAPEUTIC POTENTIAL OF AMD3100 IN PULMONARY ARTERIAL HYPERTENSION INDUCED BY
MONOCROTALINE IN RATS.
Ferraz, E. F. ; Oliveira, A. P. F. ; Barros, F. ; Riva, D. R. ; Nascimento, J. H. M.

Objectives:
Pulmonary arterial hypertension (PAH) is a syndrome characterized by increased levels in pulmonary vascular resistance, which
can be experimentally induced by monocrotaline administration, an alkaloid drug that induces toxic effects, leading to endothelial
injury. In other hand, AMD3100 a CXCR4 blocker, is frequently associated with mobilization of endothelial progenitor cells
from bone marrow and possibly to prevent the endothelial dysfunction and PAH. Thus, our aim is evaluate the therapeutic effects
of AMD3100 in PAH model induced by monocrotaline in rats.
Twenty male Wistar rats initially weighing approximately 200g were randomized in two groups: monocrotaline (MCT) and
monocrotaline plus AMD3100 (MCT+AMD3100). PAH was induced by one dose of MCT (60 mg/kg, i.p.). Two weeks after
PAH induction, the AMD3100 (1 mg/kg/day) was administered for 14 days. The body weight was measured weekly. At end of
treatment, the hypertrophy index of right ventricle (RV) was determined as the relation between mass of RV and left ventricle
plus septum. The echocardiogram parameters [cardiac output (CO), left ventricular ejection fraction (LVEF), stroke volume (SV),
left ventricular diastolic volume (LVDV)] were assessed using a high-resolution ultrasound (Vevo 770, Visualsonics). The
pulmonary airway pressure was estimated by ventilatory mechanical study. Results are expressed as mean SD, with p < 0.05 as
significative. There was no significant difference in body weight, heart weight and hypertrophy index of right ventricle. After 4
weeks, echocardiographic analysis showed decreased values of CO in MCT group compared to pre-MCT treatment (69.56
10.23 ml/min; 53.29 20.07 ml/min, p < 0.05). However, no difference was observed on CO in MCT+AMD3100 group. No
difference was found on LVEF and SV between MCT and MCT+AMD3100. The LVDV was reduced in MCT group compared
to pre-MCT treatment (407.50 86.02 l; 322.2 77.47 l, p < 0.05), but no significant difference was observed in
MCT+AMD3100 group between initial period and at end of treatment. In other hand, a significant decreased value of pulmonary
airway pressure was observed in MCT+AMD3100 group compared to MCT animals (12.25 2.63 mmHg; 18.80 3.96 mmHg,
respectively). Mortality presented a tendency to be lower in MCT+AMD3100 (37.5 %) than MCT (50%) group, but without
statistical difference.
Conclusions:
The AMD3100 treatment prevented the reduction of CO and LVDV and reduced the pulmonary airway pressure of rats with
monocrotaline-induced pulmonary arterial hypertension.
Keywords: AMD3100, CXCR4, Monocrotaline, Pulmonary arterial hypertension
Financial Support: FAPERJ, CNPq
Resumo:02-083
GHRELIN SIGNALING IN OBESE MICE HEART
Lacerda-miranda, G. ; Vieira, A. K. G. ; Sores, V. M. ; Bernardes, A. F. ; Lessa, J. G. ; Cunha, A. C. S. R. ;

Mattos, A. B. M. ; Cortez, E. ; Garcia-souza, E. P. ; Moura, A. S.
Departamento de Cincias Fisiolgicas/LFND/IBRAG, UERJ
Objectives:
Obesity is related to myocardial dysfunction and heart failure. We examined key proteins of cardiomyocyte metabolism in heart
left ventricle from obese (OG) and control (CG) groups from adult mice (180 days) overfed during lactation.
Methods: Obesity was induced by litter reduction. Therefore, the study was done in adult mice 180 days old (OG, obese
group(n=10) and CG, control group (n=10). The cardiomyocytes (cmy) of left ventricle were analyzed by light microscopy and
stereology. The content and phosphorylation of cardiac proteins: growth hormone secretagogue receptor 1a (GHSR-1a), protein
kinase B (AKT and pAKT), phosphatidil inositol 3 kinase (PI3K), AMP-activated protein kinase (AMPK and pAMPK) and actin
was achieved by western blotting. Results are expressed as mean S.E.M. Statistical significance was determined by Student ttest for unpaired. P < 0.05 was considered statistical significant. Results: Body weight (67.63 8.89g), Blood glucose (160.2
5.91g), liver weight (2.55 0.32g), and visceral fat weight (5.33 0.98g) were higher in OG (n=10) than CG group (n=10)(p
Conclusions:
Early life overnourishment induces in myocardial heart remodeling associated to increase of GHS-R1a, PI3K , AKT but not
AMPK in obese mice heart.
Keywords: ghrelin, heart, cardiomiocity, obesity, hypernutrition
Financial Support: CNPq , FAPERJ, Vital Brasil
Resumo:02-084
MODULATION OF THE UBIQUITIN PROTEASOME SYSTEM IN THYROID HORMONE-INDUCED CARDIAC
HYPERTROPHY.
Lino, C. A. ; Barreto - Chaves, M. L. M.

DEPTO. DE ANATOMIA / INSTITUTO DE CINCIAS BIOMDICAS / USP, ICB-USP
Objectives:
To evaluate the expression of proteins involved with the ubiquitin proteasome system (UPS) in thyroid hormone-induce cardiac
hypertrophy.
Male Wistar rats at the same age (~235 g) were randomized in two groups: control and hyperthyroid. Experimental
hyperthyroidism was induced by daily intraperitoneal injections of 3,5,3-triiodothyronine (7 g/100 g body weight/day ) for 21
days. Hyperthyroidism was confirmed by the ratio between heart weight (HW) and tibias length (TL). The cardiac mass
increased 31% in hyperthyroid animals compared to control (n=3; p=0,0087). The heart rate (HR) was determined during the
experimental period by tail-cuff plethysmograph (Kent Scientific, Litchfield, CT) and it was increased in the hyperthyroid
animals from 7th day of treatment (n=3; p=0,0003). Preliminary results indicated an increase on gene expression of 5 subunit of
proteasome in hyperthyroid group, evaluated by real time RT-PCR. Besides, MuRF1 expression, an E3 ligase, seems to be
augmented in the myocardium of hyperthyroid animals. Although we have observed an increase on transcription levels of some
UPS components, the expression of ubiquitinated proteins analyzed by Western blot remained unchanged between groups.
Conclusions:
Thyroid hormone exerts several effects on the cardiovascular system. Hyperthyroidism, characterized by high levels of
circulating T3, markedly stimulates the protein synthesis in the heart and leads to a concentric cardiac hypertrophy. The UPS
represents the main mechanism of degradation of intracellular cytosolic and nuclear proteins and its activation has been described
in different models of cardiac hypertrophy. The cardiac cell growth is accompanied by an increase in protein ubiquitination,
proteasome subunit expression, and proteasome activity. For instance, we observed the characteristic cardiac hypertrophy of
hyperthyroidism condition and an increase in gene expression of some components of UPS. Data regarding ubiquitinated proteins
expression and proteasome activity remain to be evaluated yet.
Keywords: SISTEMA UBIQUITINA-PROTEASSOMA, HIPERTROFIA CARDACA , HORMNIOS TIROIDEANOS
Financial Support: CAPES
Resumo:02-085
INFLUENCE OF HYPERTONIC SALINE SOLUTIONS ON HEART RATE VARIABILITY
Farah, H. M. A. T. 1; Almeida, R. L. 1; Mostarda, C. T. 2; Colombari, D. S. A. 4; Farah, V. M. A. 3; Sato, M.

A. 1; Irigoyen, M. C. 2; Colombari, E. 4
1
Depto de Morfologia e Fisiologia/ Fac. de Medicina do ABC, FMABC
2
INCOR-USP, INCOR
3
Depto de Fisiologia/ Universidade Presbiteriana Mackenzie, Mackenzie
4
Depto Patologia e Fisiologia/ Fac. Odont. Araraquara - UNESP, FOAr-UNESP
Objectives:
Central and peripheral osmoreceptor activation elicits vasopressin secretion and changes in sympathetic activity. It is still
unknown if autonomic changes can be evoked even in the absence of arterial pressure alterations. Indeed, the main objective of
this study was to investigate the acute effect of hypertonic NaCl infusion on blood pressure (BP), heart rate (HR) and
cardiovascular autonomic modulation by spectral analysis.
We used male Wistar rats weighting from 280 to 350g. The rats were divided in two groups: 1. Central infusion rats (CIR, n=5),
the rats were anesthetized with ketamine (50 mg/Kg ip) and xylazine (10 mg/Kg im) to implant a guide cannula into the lateral
ventricle (LV). After 5 days of recovery, the right femoral artery was cannulated. The solution of 2 L of 0.9%, 1.8%, 3.6% and
7.2% NaCl were infused during 2 min in the LV after 1 day of recovery. 2. Peripheral infusion rats (PIR, n=9), the rats were
anesthetized with ketamine (50 mg/Kg ip) and xylazine (10 mg/Kg im) to insert cannulas in the right femoral vein and in the right
femoral artery. Saline solutions of 0.1 mL were infused in the right femoral vein after 1 day of recovery. BP and HR were
evaluated in freely moving conscious animals through a polyethylene tubing inserted in the right femoral artery, using data
acquisition system Windaq, 2000Hz). HR variability was evaluated by spectral analysis using CardioSeries. Animals with
histological confirmation of LV cannulation were considered in this study. Statistical differences were accepted as significant at p
Conclusions:
The solutions administrated in the PIR group altered HR and its modulations while the solutions infused in the LV altered only
the BP sympathetic modulation. The volumes injected were too small (2 L for LV rats and 0.1 mL for peripheral rats) to cause
any change in HR or BP, therefore we can postulate that repeated salt exposure might induce autonomic changes even without
permanent or maintained hypertension.
Keywords: sodium chloride, heart rate, blood pressure, spectral analysis, lateral ventricle
Financial Support: PIBIC-CNPq, FAPESP and NEPAS.
Resumo:02-086
EFFECTS OF 7-DAY EXPOSURE WITH LOW DOSE OF LEAD ACETATE (PB+2) ON THE CARDIAC FUNCTION
AND MYOCARDIAL MECHANICS OF RATS.
Freire Jr, D1; dos Santos, L. 1; Fiorim, J. 1; Stefanon, I. 1; Vassallo, D. V. 1,2

1
Department of Physiological Sciences., UFES
2
Health Science Center of Vitria, Emescam
Objectives:
Lead (Pb+2) is considered an environmental pollutant of high risk to public health concerning chronic exposure. Although short
term exposures with low Pb+2 doses were rarely analyzed, we recently demonstrated effects on blood pressure and vascular
reactivity of rats. Our objective is to investigate the effects of a short term Pb+2 exposure on the cardiac hemodynamics and in
vitro myocardial mechanics from left (LV) and right (RV) ventricles of rats.
Lead acetate (Pb, n=10) or saline (CT, n=10) was injected in Wistar rats during 7 days (1st attack dose 0.4g/kg, subsequent
maintenance doses 0.005g/kg, i.m. daily). Under anesthesia (urethane 1.2g/kg, i.p.), intraventricular pressures of LV and RV
were assessed by catheterization. Thereafter, hearts were excised and LV papillary muscles and RV stripes were dissected and
placed in an isolated organ bath for in vitro study of myocardial mechanics. In vitro protocols included: steady state contractions
(Lmax); pause-dependent potentiation of force (PPP); and contractile response to isoproterenol (10-4M). For assessment of
transsarcolemmal calcium influx on the contractility, developed force were analyzed in preparations with depleted sarcoplasmic
reticulum (caffeine 10mM) after 10-minute of rest (PRC) by pause of electrical stimulus. P
Conclusions:
Although with low blood concentration, 7-day administration of Pb+2 increased arterial pressure and depressed LV function,
associated with impaired myocardial mechanics of both ventricles. Changes in cardiac muscle may include impaired inotropic
response, and depressed role of sarcoplasmic reticulum and transsarcolemmal calcium influx. Elevated RV pressure with
impaired contractility also suggests some effect on pulmonary vascular resistance.
Keywords: lead , cardiac function, MYOCARDIAL MECHANICS
Financial Support: CNPq,CAPES, FAPES
Resumo:02-087
RALOXIFENE REDUCES BLOOD PRESSURE IN HYPERTENSIVE ANIMALS AFTER OVARIAN HORMONE
DEPRIVATION.
Almeida, S. A. 1; Tiradentes, R. V. 1; Borgo, M. V. 1; Santuzzi, C. H. 1; Moraes, A. N. 1,2; Claudio, E. R. G.

1
; Endlich, P. W. 1; Gonalves, W. L. S. 1; Abreu, G. R. D. 1; Gouva, S. A. 1
1
Depto de Cincias Fisiolgicas, UFES
2
Depto de Cincias da Sade - CEUNES, UFES
Objectives:
Raloxifene displays the profile of a selective estrogen receptor modulator that could be applied as a potential preventative for
osteoporosis but with the additional benefit of preventing breast cancer and coronary heart disease. We investigated the effects of
raloxifene on blood pressure and renal excretion of water and sodium in 2-kidney-1-clip (2K1C) female hypertensive rats that had
previously been ovariectomized and their possible association with changes in endothelial function, as assessed by plasma
nitrate/nitrite determination, which is a surrogate measure of nitric oxide (NO) production.

The experimental groups were as follows: (1) hypertensive (2K1C), (2) hypertensive ovariectomized (2K1C+OVX), and (3)
hypertensive ovariectomized treated with raloxifene (2K1C+OVX+R). Seven days after the surgery that produced menopause,
2K1C hypertension was produced in anesthetized animals by applying a silver clip on the left renal artery. Seven days after the
clip application, the rats were put in metabolic cages to allow for the measurement of water ingestion and diuresis, and raloxifene
was administered (2 mg/kg/day i.p., for 7 more days). We found that the treatment with raloxifene promoted a large reduction (P
< 0.01) in blood pressure (197 6 to 164 2 mmHg), an important increase in renal excretion of sodium and water
(2K1C+OVX: 0.755 + 0.030 mEq/day and 8.7 + 0.8 mL/day vs. 2K1C+OVX+R: 1.247 + 0.076 mEq/day and 16 + 2.0 mL/day,
respectively) and an increase in plasma levels of nitrite/nitrate in 2K1C+OVX+R animals, when compared with the 2K1C, (23.4
1 vs. 14 0.5 nmol/mL; P < 0.01, respectively).
Conclusions:
These findings suggest that raloxifene exerted its antihypertensive effect, at least in part by increased excretion of sodium and
water in addition to increasing plasma levels of NO, factors that certainly with contribute to the reduction of hypertension.
Keywords: Hypertension, Ovariectomy, Raloxifene
Financial Support: CAPES and CNPq
Resumo:02-088
EFFECTS OF ORAL L-ARGININE ADMINISTRATION ON SERUM LEVELS OF NITRIC OXIDE IN
RENOVASCULAR HYPERTENSION
Tiradentes, R. V. ; Santuzzi, C. H. ; Borgo, M. V. ; Claudio, E. R. G. ; Brum, M. F. ; Endlich, P. W. ;

Abreu, G. R. D. ; Gouva, S. A.
Depto de Cincias Fisiolgicas , UFES
Objectives:
We have previously shown that L-arginine (L-Arg) has natriuretic and antihypertensive effects in renovascular hypertensive rats.
This decrease in blood pressure can be due to an increase in nitric oxide (NO) production. This work was designed to investigate
the ability of L-Arg to increase NO production in normotensive and renovascular hypertensive rats.
Male Wistar rats were anesthetized and a silver clip (0.2-mm) was placed around the left renal artery to produce the 2-kidney,
one-clip renovascular hypertension model. Another group was submitted to similar procedure and treated with L-Arg (10 mg/mL,
p.o.) from the 7th day after surgery. Sham operated rats were used as controls. At the end of the treatment, mean arterial pressure
(MAP) was measured in conscious animals. Serum NO2-/NO3- levels were measured in hemoglobin-free plasma-derived serum
(PDS) by breaking the clot and filtered it through a 10 KDa cut-off membrane and centrifuged on a fixed-angle rotor at 10,000
rpm for 1 hr at 4XC. Nitrate reductase was added to the samples to convert NO3- to NO2-. Total NO2- was then analyzed by
reacting the samples with the fluorometric reagent 2,3-diaminonaphthalene and measuring emission at 375 nm and excitation at
415 nm. Nitrite was taken as a reliable index of NO production. We observed a significant reduction in MAP of L-Arg-treated
when compared to untreated rats (120b5 vs. 170b8 mm Hg) and, concurrently, occurred an increase in NO production
(25.64.8 vs. 11.81.5 nmol/mL). The treated and untreated sham groups did not differ at the fluorometric assay (14.52 and
13.92.7 nmol/mL and MAP measurements (1023 and 1043 mm Hg)
Conclusions:
These results suggest that L-Arg treatment has an stimulatory action on the systemic NO formation, which probably explains its
hypotensive and natriuretic effects in renovascular hypertensive rats.
Keywords: L-arginine, Nitric Oxide, Renovascular Hypertension
Financial Support: CAPES and CNPq (Brazil)
Resumo:02-089
EVALUATION OF CARDIOVASCULAR PARAMETERS OF FEMALE RAT OFFSPRING EXPOSED TO THE
EXTRACT OF ST. JOHN'S WORT
Hamada, R. Y. ; Cunha, N. V. D. ; Reis, D. A. S. ; Pinge, M. C. M. ; Gerardin, D. C. C. ; Gomes, G. G. P.

UNIVERSIDADE ESTADUAL DE LONDRINA, UEL
Objectives:
Psychiatric disorders in pregnancy are quite common and the use of psychotropic drugs during these periods is, in most cases,
indispensable. An effective alternative to synthetic antidepressants is the extract of Hypericum perforatum (HP), St. John's Wort,
which can increase levels of serotonin. Studies suggest that antidepressants may interfere with the cardiovascular system of
depressed patients. The widespread use of the extract of HP has attracted attention from the scientific community about its use
during pregnancy and lactation with possible consequences in the offspring, because the herbal drugs are considered safe and
popular self-medicated without professional supervision. Previous study of our laboratory demonstrated that the use of HP in rats
during pregnancy and breastfeeding may increase the basal heart rate in the male offspring in adulthood. In the present work, we
aimed to verify the baseline cardiovascular parameters of female rat offspring exposed to the extract of HP.
Wistar rats (n= 7) were treated daily with 100 mgkg of HP or 0.3 ml of 0.9% saline solution (control group) from the first day of
pregnancy (GD1) to the 21st day after birth of pups (PND 21, weaning). After 90 days, immediately prior to starting the
experimental preparation, a vaginal smear was taken from female offspring to determine the stage of the estrous cycle and only
the females presenting estrus phase were used to evaluated cardiovascular parameters. These females underwent surgery to
implant a catheter in the femoral artery for recording of blood pressure and heart rate. The maternal treatment with HP did not
change baseline mean arterial pressure (MAP) and heart rate (HR) of female pups (MAP=110 1.5 mmHg and HR: 400 22
bpm, n=6) when compared with the control group (MAP=113 1.4 mmHg and HR: 393 12 bpm, n=11).
Conclusions:
The results demonstrate that the use of HP in rats during pregnancy and breastfeeding did not modify the basal cardiovascular
parameters in the female offspring in adulthood.
Keywords: CARDIOVASCULAR , EXTRACT OF ST. JOHN'S WORT, HYPERICUM PERFORATUM, FEMALE RAT
Financial Support: PIBICUEL
Resumo:02-090
INVOLVEMENT OF INOS PATHWAY INTO THE PARAVENTRICULAR NUCLEUS OF THE HYPOTHALAMUS
(PVN) IN CARDIOVASCULAR AND AUTONOMIC MODULATION DURING LPS ENDOTOXEMIA IN
CONSCIOUS RATS
Matsumoto, A. K. ; Silva, A. M. D. ; Abreu, S. B. D. ; Filhho, P. P. ; Pinge, M. C. M.

Cincias Fisiolgicas/Universidade Estadual de Londrina, UEL
Objectives:
During sepsis, areas of the central nervous system are activated and have a role in physiological adaptations to immunity
challenge. One of these areas is the paraventricular nucleus of the hypothalamus (PVN). Data from our laboratory showed the
involvement of PVN in cardiovascular and autonomic modulation during endotoxemia by lipopolysaccharide (LPS). The PVN is
also an important site where nitric oxide (NO) plays a relevant role.The aim of this study was to evaluate the involvement of
nitric oxide from iNOS pathway in the PVN on mean arterial pressure (MAP), heart rate (HR) and spectral analysis of HR
variability (HRV) after administration of LPS in conscious rats.
Male Wistar rats were anesthetized for implantation of bilateral guide cannulae to the PVN. After 5 days the rats were
anesthetized for chronic catheterization of the femoral artery and vein. 24 hours later it was performed the bilateral microinjection
of 100 nl of saline or aminoguanidine (250 pmol) into the PVN of conscious rats, followed by LPS administration (iv).
Cardiovascular parameters were analyzed by 2 hours. Saline microinjection into the PVN did not modify the baseline parameters
and after LPS was observed hypotension and tachycardia ( MAP= -24 5 mmHg, HR = 85 15 bpm, p
Conclusions:
Our data suggest an involvement of iNOS pathway into the PVN in cardiovascular and autonomic modulation during the initial
phase of endotoxemia by LPS.
Keywords: AUTONOMIC MODULATION , ENDOTOXEMIA, PARAVENTRICULAR NUCLEUS
Financial Support: PIBIC/UEL.
Resumo:02-091
EFFECTS OF PROTEIN RESTRICTION POST-WEANING ON MORPHOLOGY AND CONTRACTILE
FUNCTIONS IN CARDIOMYOCYTES OF FISCHER RATS
Penitente, A. 1; Novaes, R. D. 1; Natali, A. J. 1; Silva, M. F. D. 1; Ribeiro, M. F. 3; Fernandes, K. M. 1;

Souza, A. M. A. D. 3; Silva, M. E. D. 3; Chianca-jr, D. A. 3; Neves, C. A. 1
1
Departament of General Biology, UFV
4
Departament of Nutrition , UFOP
3
Departament of Biological Science, UFOP
2
Departament of Physical Education , UFV
Objectives:
The present study evaluated the effects of severe protein restriction after weaning on the morphology and contractile function of
isolated left ventricular (LV) cardiomyocytes of rats.
Adult Fischer rats were randomized into control (CG = 20) and malnourished (MG = 20) groups. After weaning, the GC and MG
rats received isocaloric diets containing 15% and 6% protein, respectively, during 35 days. Subsequently, the animals were
weighed, sacrificed and the hearts were removed and weighted. LV were dissected, weighed and the cardiomyocytes were
isolated for morphological and contractile function analysis at baseline, in response to changes in extracellular calcium
concentration. Removed hearts were fixed in formalin for 48 hours and the fragments of the LV were dehydrated and embedded
in glycol methacrylate resin (Historesin, Leica) and paraffin. Serial sections were performed two and four m thick, mounted on
slides and stained with toluidine blue 1% borax, hematoxylin / eosin and subjected to histological and morphometric analysis
using an optical microscope (BX-60, Olympus) connected to a Q-Color 3 digital camera (Olympus). Area and nuclei of
cardiomyocytes were measured using Image-Pro Plus 4.5 (Media Cybernetics). Statistical analysis was performed using mean
SEM. The animals in the MG group had reduced body mass, heart and ventricular rate, but higher LV / body weight ratio when
the cardiac mass was normalized in relation to body mass. Regarding the morphological properties of the LV, the protein
restriction reduced cellular morphological parameters of cardiomyocytes analyzed, except the length / width ratio that was greater
in these MG rats. Analysis of cellular contractility demonstrated that protein malnutrition caused a reduction in the amplitude of
ventricular cardiomyocytes contraction, increased the time to peak contraction and to half relaxation of these cells. In addition,
cardiomyocytes from malnourished animals showed changes in contractile properties in response to different variations of
extracellular calcium compared to control animals.
Conclusions:
The severe protein restriction leads to changes in morphology and contractile dysfunction in left ventricular cardiomyocytes of
Fischer rats.
Keywords: cardiomyocytes, Fischer rats, protein restriction
Financial Support: FAPEMIG, CNPq, CAPES, FUNARBE, UFV
Resumo:02-092
BLOCKADE OF 5-HT3 RECEPTORS IN THE MEDIAL SEPTAL AREA CHANGES BAROREFLEX SENSITIVITY.
Urzedo-rodrigues, L. S 1; Ferreira, H. S. 2; Almeida, D. O1; Batista, A. S. 1; Dias, D. P. M. 3; Fregoneze, J.

B. 1
1
Universidade Federal da Bahia, Dpto de Fisiologia , UFBA
2
Universidade do Estado da Bahia, Dpto de Cincias da Sade, UNEB
3
Universidade de So Paulo, Dpto de Fisiologia, USP
Objectives:
It has been reported that different serotonin receptors are involved in the brain control of cardiovascular function. Recent data
from our laboratory show that 5-HT3 receptors in the medial septal area (MSA) exert a tonic inhibitory influence on blood
pressure (Autonomic Neurosc., 159:51-61, 2011). Among the several serotonin receptors, the 5-HT1A and 5-HT7 receptors seem
to be crucial for parasympathetic control of heart rate. However it is not fully understood the role of serotonin receptors on the
control of baroreflex. In the present study, we investigated whether the blockade of 5-HT3 receptors in the medial septal area
(MSA) modifies baroreflex sensitivity in rats.
Male Wistar rats (280-310g) were anaesthetized with Ketamine/Xylazine (80/11.5 mg/kg i.p) for implantation of guide cannulas
in the MSA 5 days before experiments. A catheter was inserted into the left carotid artery 24h before the experiment to allow
blood pressure recording. At the experimental session, after 30 min of baseline blood pressure recording, rats received
microinjection of 5-HT3 receptor antagonist, ondansetron (160 nmol), or 0.9% saline into MSA. At the end of the experiments,
animals were anesthetized and subjected to transcardiac perfusion with saline followed by 10% formalin and had their brain
removed for histological analysis. Only data from animals whose guide cannulas were in the MSA were considered. The
baroreflex sensitivity (BRS) was assessed in the time-domain by means of the Sequence technique. A computer software
(Analyzer v4.4) scanned beat-by-beat time series of systolic blood pressure (SBP) and inter-beat interval (IBI) searching for
sequences of at least 4 consecutive beats in which increases in SPB were followed by IBI lengthing (up sequence) and decreases
in SPB were followed by IBI shortening (down sequence). The slope of the linear regression lines between SPB and IBI was
taken as a measure of BRS. The average of the slopes, number of up and number of down sequences were calculated for each rat
at basal period (before MSA injection), at 0-30 min and 30-60 min period after MSA injections. The data were subjected to oneway analysis of variance (ANOVA) followed by Student-Newman-Keuls post-test. Ondansetron-treated rats showed a significant
reduction in the slope (1.70 0.13 ms/mmHg, n=5) of up sequences as compared to control group (3.25 0.61 ms/mmHg, n=4)
at 30-60 min. The number of up and down sequences was found similar between ondansetron-treated and saline-treated rats.
Conclusions:
The blockade of 5-HT3 receptors in the MSA reduces the average slope of up sequences. It has been shown that up
sequences are better predictors of vagal cardiac activity than down sequences. We hypothesize that 5-HT3 serotonin receptors
in the MSA are involved in the modulation of parasympathetic component of baroreflex function.
Keywords: Baroreflex Sensitivity, 5-HT3, Medial Septal Area, Serotonin
Financial Support: Capes, CNPq, FAPESB
Resumo:02-093
CARDIAC ELECTRIC REMODELLING IN RATS CHRONICALLY TREATED WITH ANABOLIC STEROID: LTYPE CALCIUM CURRENT CHARACTERIZATION
Arantes, P. C. ; Rodrigues Jr, L. F. ; Medei, E. ; Nascimento, J. H. M.

Objectives:
Cardiac abnormalities, arrhythmias and sudden death were reported as consequence of abusive use of anabolic androgen steroids
(AAS). In a previous study, we showed the increase in ventricular action potential duration (APD) in hearts of rats chronically
treated with decanoate nandrolone (DECA). The purpose of present study was to evaluate the contribution of L-type calcium
current (ICa,L) to APD prolongation on AAS-induced cardiac electrical remodelling.

Methods: Male wistar rats were treated with nandrolone decanoate (DECA group; dose: 10 mg/kg/week, i. m.) or vehicle
(Control group) during 8 weeks. Ventricular cardiomyocytes were dissociated using colagenase and ionic currents were recorded
using whole cell patch-clamp technic. Voltage protocol to obtain curve I-V consisted of 300 ms pulses, from holding potential of
-50 mV to potentials from -60 to +60 mV potentials, in 10 mV intervals. Protocol to obtain steady-state inactivation voltage
dependency consisted of 1000 ms pre-pulses from -80 mV to +40 mV, in 10 mV intervals, followed by a 300 ms pulse to 0 mV.
Data are presented as mean standard error. Results: Current amplitude was normalized by cell capacitance (Control: 108.9
9.75 pF, N = 11; Deca: 103.2 7.75 pF, N = 10). The peak density of ICa,L in Control group was -10,16 0,87 pA/pF (N = 11),
with peak current occurring at 10 mV, activation in -20 mV and reversal potential next to -40 mV. In DECA group, the peak
density of ICa,L was higher (-13,46 1,81 pA/pF; P = 0,0438 vs. Control), with peak at 10 mV. Activation V of ICa,L was -8,0
0.05 mV in Control group and -7,0 0.05 mV in DECA group, while inactivation V of ICa,L was -25 0.05 mV (Control)
and -22 0.33 mV (DECA).
Conclusions:
Our preliminary results suggest an increased density of ICa,L in left ventricle of rats chronically treated with anabolic steroids.
Keywords: Anabolic androgen steroids, Cardiac electric remodelling, L-type calcium current, Rats chronically treated
Financial Support: FAPERJ and CNPq/PIBIC
Resumo:02-094
INFLUENCE OF CRONIC CAROTID AND AORTIC BARORECEPTORS DENERVATION ON AUTONOMIC
MODULANTION IN SPONTANEOUSLY HYPERTENSIVE RATS
Souza, P. R. M. D. 1; Moreira, E. D. 1,1; Mostarda, C. 1; Monteiro-de-moraes, W. M. A. 2; Guimares, F. 2;

Santos, F. 1; Piratello, A. C. 3; Silva, M. B. 1; Irigoyen, M. C. 1
1
Experimental Hypertension Laboratory - Heart Institute (InCo, FMUSP - InCor
2
University of So Paulo Medical School, So Paulo, Brazil; S, EEFE-USP
3
PPG Nephrology, Department of Medicine Nephrology Division, UNIFESP
Objectives:
To evaluate the participation of the autonomic nervous system in hemodynamic variables through selective denervation of carotid
and aortic receptors in spontaneously hypertensive rats (SHR) after 10 weeks.
SHR (280-320g) were divided into three groups: control (N=12), aortic denervation (SHRAD, N = 13) and carotid denervation
(SHRCD, N =11). Ten weeks after the denervation procedure, the animals were catheterized (right femoral artery and vein) for
direct recording of arterial pressure (AP) and drug administration respectively. The obtained signals were later analysed by
spectral analisys. The results were reported as means SD.Results: The blood pressure, heart rate and HRV were similar between
groups. BPV increase in SHRAD when compared with SHR and SHRCD (SHRAD= 135.262 vs. SHR=70.741 and
SHRCD=61.727 mmHg2). LF and HF absolute components of HRV as well as the sympatho-vagal balance were similar
between groups. The absolute LF component of systolic blood pressure increases in SHRAD when compared with SHR
(SHRAD=18.510 vs SHR=9.25 mmHg2) However SHRCD was similar among the groups. (SHRCD=11.1 7 mmHg2)
Conclusions:
The data showed that sympathetic component of heart rate variability are preserved in SHR submitted to chronic selective
denervation. However the sympathetic component acting on the vessels was increased only in aortic denervation, showing the
differential role of the aortic baroreceptors in the homeostasis of blood pressure even in the presence of hypertension
Keywords: selective denervation, autonomic modulantion, hypertension
Resumo:02-095
TREATMENT WITH ATENOLOL (B-ADRENERGIC ANTAGONIST) ALTERS THE PROFILE
IMMUNOHISTOCHEMISTRY OF RANK, RANKL, OPG AND MMP-9 OF ALVEOLAR BONE HEALING PROCESS
IN SPONTANEOUSLY HYPERTENSIVE RATS (SHR)
Manrique, N. ; Pereira, C. C. S. ; Okamoto, R. ; Antoniali, C.

Faculdade de Odontologia de Araatuba-UNESP, FOA-UNESP
Objectives:
Previously results obtained in our laboratory showed that there is a delay in alveolar bone healing process in SHR. This delay was
associated with increased peripheral sympathetic tone of these animals, since treatment with atenolol reduced SBP (systolic blood
pressure) and normalizes alveolar bone formation. The aim of this study was to evaluate the atenolol effect on the expression of
bone metabolism proteins OPG, RANK, RANKL and MMP-9 in dental alveoli of SHR
Wistar rats and SHR treated or not with atenolol (100mg/kg/d, po) were subjected to surgical extraction of upper right incisor
tooth. Treatment with atenolol was started one week before surgery and maintained until sacrifice. The alveolar repair was
evaluated in the different phases: initial (7 day), bone formation (14 and 21 days) and remodeling (28 and 42 days). The upper
right maxilla were processed and submitted to immunohistochemical reactions. The middle third of dental alveolus was examined
(20x objective), staining and it was analyzed qualitatively by assigning scores. The results were compared between groups (Mann
Whitney, p
Conclusions:
Our results demonstrated that: 1- RANKL, RANK and MMP-9 but not OPG, are associated with the delay of the alveolar bone
healing process in SHR, 2- treatment with atenolol increases the OPG staining in all phases analyzed and RANKL, RANK and
MMP-9 specifically in 21 e 42 days. These results suggest that in SHR, changes in the initial phase of alveolar bone formation
and remodeling would be a consequence of a differential expression of proteins in bone metabolism such as OPG, RANKL,
RANK and MMP-9, which is modulated by treatment with atenolol.
Keywords: Alveolar process, Atenolol, Hypertension, Immunohistochemistry, Rats, inbread SHR
Financial Support: Fapesp (2008/01893)
Resumo:02-096
REGIONAL EFFECTS OF RESISTANCE-TRAINING ON CELL SIZE IN RAT CARDIAC MYOCYTES
Melo, S. F. S. ; Junior, M. A. C. ; Bozi, L. ; Drummond, L. R. ; Natali, A. J. ; Oliveira, E. M. D.

Escola de Educao Fsica , USP
Objectives:
The purpose of this study was to test whether the hypertrophic response to resistance-training program for a period of 2 months
was uniformly distributed across the ventricular wall in rats.
Fourteen animals were randomly divided into two groups: control (n = 6, CO), and trained (n = 7, TR). The training protocol
consisted of four sets of 1012 repetitions of the squat exercise performed at 7580% of one repetition maximum (1RM). Single
ventricular myocytes were isolated from the sub-epicardium (EPI) and sub-endocardium (ENDO) of trained rats and from
sedentary rats for comparison. Cellular hypertrophy (approximately 14% greater cell volume, P
Conclusions:
We have previously shown using this model that resistance training induces the development of concentric cardiac hypertrophy
without ventricular dysfunction or cavity reduction. Here, ours results showed difference between EPI and ENDO
cardiomyocytes size to trained compared to the sedentary rats. Also, to the proteins involved in the maintenance of normal
cardiac Ca2+ homeostasis improving the contractile function in trained group compared to the sedentary.
Keywords: CARDIAC MYOCYTES, RESISTANCE-TRAINING , MYOCYTES
Financial Support: FAPESP
Resumo:02-097
PHOSPHODIESTERASE INHIBITION INCREASES THE POTENCY OF RELAXATION OF THE NEW NO-DONOR
TERPY IN OLD SHR.
Munhoz, F. C. 1; Potje, S. R. 1; Hildebrand, M. C. 1; Pereira, A. C. 2; Ramos, L. C. B. 2; Bendhack, L. M. 2;

Antoniali, C1
1
Dept of Basic Sciences, School of Dentistry of Araatuba, FOA-UNESP
2
School of Pharmaceutical Sciences, FCFRP-USP
Objectives:
The vasodilatory effect of TERPY, a new NO donor drug, is more potent in old than in young spontaneously hypertensive rats
(SHR). These results suggest that the mechanisms involved in the effect of TERPY could be modulated by age in SHR. So, the
aim of this study was to evaluate possible alterations in the action of soluble guanilate ciclase (sGC), potassium channels (K+),
phosphodiesterase and superoxide anion (O2-) involved in vasodilatory effect of TERPY in old SHR.
Animal Research Ethics Committee (CEEA) at the School of Dentistry of Araatuba approved all the experimental protocol
conduced in the study (proc. number 001776-2010). 4 months old WST and SHR were used in the young group and animals with
16 months were considered old. These animals were killed and had the aorta removed and isolated. Aortic rings were placed
between two stainless-steel stirrups and connected to an isometric force transducer (Letica Scientific Instruments; Barcelona
Spain) to measure isometric tension in the vessels. The rings were placed in an organ chamber containing Krebs solution
maintained at pH 7.4 gassed with 95% O2 and 5% CO2 at 37C. The rings were initially stretched to a tension of 1.5 g and then
they were allowed to equilibrate for 60 min. Denuded aortic rings from SHR (SP> 150mmHg) and WST were pre-contracted
with phenylephrine and concentration-response curves for TERPY (1nM- 100 M) were constructed in the presence or absence
of ODQ (GCs inhibitor, 1M), TEA (K+ channel blocker, 1mM), DIPYRIDAMOLE (Phosphodiesterase inhibitor, 1M),
TIRON (O2- scavenger, 0.1mM) and APOCYNIN (NADPH oxidase inhibitor, 0.1mM). Curves were normalized in percentage of
relaxation, according to the level of contraction. Potency (pD2) and efficacy (maximum effect, ME) of TERPY were evaluated.
The results (meanSEM) were compared between the groups by Students t test (p
Conclusions:
Taken together, these results show that TERPY induced relaxation is GCs dependent and this dependency is more evident in old
SHR. K+ channels sensitive to TEA and superoxide anions scavenged by TIRON do not to interfere in the effect of the
compound. Besides, in old SHR aorta, NADPH oxidase inhibition doesnt increases the potency of TERPY as in young SHR, but
the increased potency in the phosphodiesterase inhibition by DPYRIDAMOLE was more evident in young SHR. These data
suggest that the participation of NADPH oxidase and phosfodiesterase in the relaxing effect of TERPY is differently modulated
by age in SHR.
Keywords: Nitric Oxide donors, SHR, advanced age, phosphodiesterase, NADPH oxidase
Financial Support: CNPQ
Resumo:02-098
EFFECTS OF ADRENOMEDULLIN ALONE OR COMBINED WITH GLUTAMATE IN THE ROSTRAL
VENTROLATERAL MEDULLA OF CONSCIOUS RATS
Barbosa, R. M. ; Barbosa, S. P. ; Freiria-oliveira, A. H. ; Menani, J. V. ; Colombari, E. ; Colombari, D. S.

A.
Dept of Physiology and Pathology, FOAr-UNESP
Objectives:
It has been demonstrated that the peptide adrenomedullin (ADM) microinjected into the rostral ventrolateral medulla (RVLM) of
anesthetized rats produces increases in arterial pressure and in the sympathetic activity, an effect that seems to be mediated by
glutamatergic receptors (Am. J. Physiol., 287: R729, 2004). A previous study from our laboratory demonstrated that the pressor
response induced by glutamate injected into the RVLM was enhanced by the pre-treatment with the peptide angiotensin II also in
the RVLM (Brain Res., 1322: 72, 2010). Due to this interaction, our hypothesis is that ADM in the RVLM could also increase the
pressor response induced by glutamate in the RVLM. Since the anesthesia can alter the response of the substances injected in the
central nervous system and to further study the interaction between ADM and glutamate in the RVLM, in the present study we
investigated the effects of ADM alone or combined with glutamate into the RVLM on the arterial pressure in conscious rats.
Male Holtzman rats (280-320 g) with stainless steel guide cannulas unilaterally implanted in the RVLM (n = 5-8/group) were
used. In the day before the experiments, catheters were inserted into the femoral artery for mean arterial pressure (MAP)
recording in conscious animals. In one group of rats glutamate (1 nmol/100 nl) was injected before and 5 min after ADM (100
pmol/100 nl) and in another group of rats glutamate (1 nmol/100 nl) was injected before and 20 min after ADM (100 pmol/100
nl). ADM into the RVLM increases MAP (15 4, vs. saline: -1 1 mmHg, p < 0.05). The pressor responses induced by
glutamate into the RVL before and 5 min after ADM were similar (35 6, vs. after ADM: 36 6 mmHg). In another group of
rats, ADM also induced pressor response (20 5 mmHg, vs. saline: -2 1 mmHg, p < 0.05) and, similarly, there was no
difference between the pressor responses induced by glutamate into the RVLM before (24 7 mmHg) and 20 min after ADM (24
5 mmHg). Baseline MAP was 121 4 and 117 5 mmHg, for each group of rats.
Conclusions:
ADM microinjected into the RVLM of conscious rats also induces increases in MAP, as observed in anesthetized rats. Different
from our hypothesis, ADM did not change the pressor response to glutamate into the RVLM, even shortly after ADM, suggesting
that in conscious rats the interaction between ADM and glutamate at the level of RVLM may not occur.
Keywords: blood pressure, RVLM, glutamatergic pathway
Financial Support: PIBIC-CNPq, FAPESP
Resumo:02-099
ROLE OF ANGIOTENSIN II AT1 RECEPTOR IN THE ANGIOTENSIN-(1-7)-INDUCED CORONARY
VASODILATION IN HYPERTROPHIED RAT HEARTS.
Almeida, J. F. Q. ; Sobrinho, D. B. S. ; Alves, G. M. M. ; Dourado, J. G. ; Macedo, L. M. ; Mendes, E. P. ;

Castro, C. H.
Depto. de Cincias Fisiolgicas / Uni. Federal de Gois, ICB
Objectives:
Cardiac hypertrophy is an adaptive response of the heart associated with pathological situations. Pressure overload-induced left
ventricular hypertrophy (LVH) is related with impaired coronary circulation. Our previous study showed that Angiotensin-(1-7)
[Ang-(1-7)] and the receptor Mas are involved in the coronary circulation in physiological conditions. The aim of this study was
to evaluate the involvement of the angiotensin receptors on the coronary effect of Ang-(1-7) in hypertrophied hearts.
Subtotal abdominal aortic banding (AB) was performed to induce left ventricular hypertrophy in rats. After 21 days, the hearts
were perfused according to the Langendorff method (constant flow). After a basal period, the hearts were perfused for 15 min
with Ang-(17) (20 pmol/L) in presence or absence of receptor Mas antagonist A-779 (2 nmol/L), AT1 antagonist, losartan (1
mol/L), or in combination. The left ventricles wet weights were recorded, normalized for tibia length and then expressed as
ventricular mass index. Aortic constriction resulted in a significant left ventricular hypertrophy (0,174 0,004 vs
0,2170,012g/cm in AB hearts, p
Conclusions:
These findings indicate that Angiotensin II AT1 receptor is able to modulate Ang-(1-7)-induced coronary vasodilation in
hypertrophied rat hearts.
Keywords: A779, ANGIOTENSIN-(1-7), cardiac hypertrophy, LOSARTAN, receptor MAS
Financial Support: CNPq/INCT-Nanobiofar.
Resumo:02-100
SEROTONIN UPTAKE IN JUGULAR VEIN AND THE INVOLVEMENT OF MONOAMINE TRANSPORTERS.
de Pr, M. A. ; Poli, A. ; Linder, A. E.

FARMACOLOGIA, UFSC
Objectives:
Serotonin (5-hydroxytryptamine; 5-HT) can interact with its receptors causing vascular contractility in a variety of vessels,
including arteries and veins. In the rat jugular vein, there is evidence that the 5-HT2A receptor mediates contraction (JPET 334;
116, 2010). The presence of the serotonergic system in the peripheral vasculature enables the uptake and metabolism of this
monoamine by arteries and veins. 5-HT uptake in veins is 5-HT transporter (SERT) independent (JPET 323; 415, 2007).
Nowadays, substances that interfere in the uptake and metabolism of monoamines are widely used to treat neurological disorders,
although little is known about their influence on the cardiovascular system. The aim of this work is to test the hypothesis that 5HT uptake in veins is mediated by the norepinephrine (NE) transporter and that its function modulates venous reactivity.
Jugular veins isolated from male Wistar rats (300-400 g) were placed and maintained in physiological salt solution (PSS) at 370C
containing either vehicle or 5-HT (1 uM) for 15 min. 5-HT (1 uM) was also added for 15 min to vessels that had been previously
exposed for 30 min to the inhibitor of 5-HT uptake, fluoxetine (1 uM), or to the mixed 5-HT and NE uptake inhibitor, imipramine
(1 uM). Tissues were dipped several times in drug-free PSS to avoid extracellular 5-HT contamination, and then processed for
posterior measurements of 5-HT and its metabolite 5-hydroxy indol acetic acid (5-HIAA) by HPLC. Data were expressed as
nanogram of 5-HT or 5-HIAA per milligram of wet tissue. For isometric tension recordings, jugular vein rings were isolated and
mounted in an organ bath. Concentration-effect curves to 5-HT (1 nM 0.1 uM) were performed in the presence and absence of
imipramine (1 uM) or fluoxetine (1 uM). Experimental protocol approved by the ethical committee (p0036).Results: Basal levels
of 5-HT could be measured in jugular vein (6,12 2,50 ng/mg n=5), whilst the main product of its metabolism 5-HIAA was not
detectable. Whereas no significant changes in 5-HT content (6,28 1,16, n=5; P>0.05) were detected when compared to basal
levels, a significant increase in 5-HIAA content was observed after 15 min (1,88 0,47 ng/mg n=5) of extracellular 5-HT
exposure, indicating 5-HT uptake followed by metabolism. Fluoxetine had no effect in 5-HT and 5-HIAA content in jugular vein
segments exposed to 5-HT, as expected (P > 0.05). Interestingly, when compared to 5-HT exposure for 15 min alone, imipramine
decreased 5-HT levels (2,77 0,73 ng/mg), but not the 5-HIAA levels. The concentration-effect curves to 5-HT were not altered
in the presence of imipramine or fluoxetine (p>0,05).
Conclusions:
Our results support previous findings that 5-HT uptake is SERT-independent in veins. Moreover, we present evidence of a
putative role for the NE transporter in the uptake of 5-HT in the rat jugular vein.
Keywords: SEROTONIN, UPTAKE, VASCULAR REACTIVITY, VEIN
Financial Support: CNPq, FAPESC/CNPq (005/2009), Funpesquisa.
Resumo:02-101
REDUCTION OF VENTRICULAR DYSFUNCTION INDUCEDBY MYOCARDIAL INFARCTION BY HYDRO
ALCOHOLIC EXTRACT OBTAINED FROM FRUITS OF EUTERPE OLERACEA MART. (AAI)
Silva, J. S. D. 1; Zapata-sudo, G. 1; Seabra, S. 4; Sousa, P. J. D. C. 2; Resende, A. C. 3; Moura, R. S. 3; Sudo,

R. T. 1
1
Programa de Desenvolvimento de Frmacos - ICB, UFRJ
2
Departamento de Farmacologia, UFPa
3
Departamento de Farmacologia, UERJ
4
Departamento de Farmacologia, UEZO
Objectives:
Delay in the progress of cardiac failure in cardiovascular diseases such as myocardial infarction (MI) could be observed if cardiac
remodeling processes are interrupted. The purpose of this study was to investigate cardiac remodeling and function in rats
submitted to experimental MI and treated with an extract, obtained from seed of aai fruits (Euterpe oleracea Mart) that have
antioxidant, vasodilation and anti inflammatory actions.
Protocols were approved by Animal Care and Use Committee at Universidade Federal do Rio de Janeiro. Wistar male rats (180200 g) were randomly divided in sham-operated (SHAM) and infarcted groups (MI) and each group subdivided in: treatment with
vehicle and with aai (100 mg/kg, p.o.). Under sevoflurane anesthesia, the animals were submitted to a ligature of the anterior
descending coronary artery. After 28 days of treatment, the following hemodynamic parameters were analyzed: mean arterial
pressure (MAP), left ventricular end diastolic pressure (LVEDP), end-systolic pressure (LVESP) and dP/dt. Hypertrophy was
determined using the ratio of heart/body weight. Results- The following table summarizes the results obtained in both SHAM and
MI groups treated or non-treated with the extract. Hemodynamic and heart weight change induced by myocardial infarction (MI)
and treatment with Euterpe oleracea. Mean arterial pressure (MAP), left ventricular end-systolic pressure (LVESP), left
ventricular end diastolic pressure (LVEDP), positive (+) and negative (-) dP/dt. Data were expressed as mean SEM (n= 6). *P
Conclusions:
Our results demonstrate that aai fruit extract has a significant protection on cardiac dysfunction induced by MI. This beneficial
action of aai is probably dependent on the reduction of myocardial fibrosis due to a protective action against oxidative stress,
reduction of inflammatory reaction and an increase in vascular blood flow to the area of infarction.
Keywords: Aai, Antioxidant, Cardiac failure, Myocardial Infarction, Ventricular dysfunction
Financial Support: FAPERJ, Capes, CNPq, INCT/INOFAR
Resumo:02-102
NADPH OXIDASE INHIBITION POTENTIATES THE RELAXATION INDUCED BY ([RU(TERPY)(BDQ)NO+]3+)
TERPY IN SHR.
Potje, S. R. 1; Munhoz, F. C. 1; Hildebrand, M. C. 1; Ramos, L. C. B. 2; Bendhack, L. M. 2; Antoniali, C. 1

1
Department of Basic Sciences, FOA-UNESP
2
Dept of Physics and Chemistry, FCFRP-USP
Objectives:
Previous experiments show that the hypotensive effect of TERPY, a new NO donor, is increased in SHR, compared to Wistar rats
(WST). Our hypothesis is that this effect could be associated to an increase in the relaxing response of vascular smooth muscle
induced by TERPY. Besides, oxidative stress could be differently modulated in SHR. So, the aim of the present study was to
evaluate the effect of TIRON, a superoxide anion scavenger, and APOCYNIN, a NADPH oxidase inhibitor in the relaxation
induced by TERPY in SHR aorta.
Animal Research Ethics Committee (CEEA) at the School of Dentistry of Araatuba approved all the experimental protocol
conduced in the study (proc. number 001776-2010). 4 months old WST and SHR were killed and had the aorta removed and
isolated. Aortic rings were placed between two stainless-steel stirrups and connected to an isometric force transducer (Letica
Scientific Instruments; BarcelonaSpain) to measure isometric tension in the vessels. The rings were placed in an organ chamber
containing Krebs solution maintained at pH 7.4 gassed with 95% O2 and 5% CO2 at 37C. The rings were initially stretched to a
tension of 1.5 g and then they were allowed to equilibrate for 60 min. Endothelium integrity was assessed by the degree of
relaxation induced by acetylcholine in phenylephrine pre-contracted arteries. Denuded aortic rings from SHR (SP> 150mmHg),
and WST were pre-contracted with phenylephrine and concentration-response curves for TERPY (1nM- 100 M) were
constructed in the presence or absence of TIRON (0.1mM) and APOCYNIN (0.1mM). Curves were normalized in percentage of
relaxation, according to the level of contraction. Potency (pD2) and efficacy (maximum relaxation, ME) of the vasodilator effects
of TERPY were compared. Differences were considered significant when p
Conclusions:
These data suggest that superoxide anions scavenged by TIRON, in the concentration of 0.1mM, dont play a significant role in
the vasorelaxing effect of TERPY in SHR. On the other hand, NADPH oxidase inhibition by APOCYNIN increases the potency
of TERPY in SHR.
Keywords: NITRIC OXIDE DONORS, SHR, OXIDATIVE STRESS, TERPY, SODIUM NITROPRUSSIDE
Financial Support: CNPq, FAPESP
Resumo:02-103
BRADYCARDIA AND PRESSOR RESPONSES TO OBSTRUCTIVE APNEA ARE REDUCED DURING REM SLEEP
IN RATS
Rossi, M. V. ; Schoorlemmer, G. H. ; Cravo, S. L.

Depto. de Fisiologia, Universidade Federal de So Paulo, UNIFESP
Objectives:
Obstructive sleep apnea is a very common condition that leads to several health problems. However, it is still not clear if the
consequences of sleep apnea are due directly to apnea, or to sleep fragmentation. We compared cardiorespiratory responses
induced by obstructive apnea in awake and sleeping rats.
We used four Wistar rats with a balloon-tipped catheter implanted in the trachea. Apnea was induced by injection of ~15 L
water into the balloon. Apnea terminated on withdrawal of the injected fluid. Balloon inflation did not cause tracheal pain
because the balloon was contained in a rigid Teflon tube. Rats also had ECG, EEG and EMG electrodes. Sleep stages were
classified online once a second according to Louis et al. (J Neurosci Methods 133: 71-80, 2004) with Spike2 software. Apnea was
initiated automatically when 4 of the last 5 seconds were classified as REM, and terminated when EMG activity increased to the
awake level. Some rats also had arterial catheters for measurement of arterial pressure, and thoracic balloons for measurement of
respiratory effort. One week after surgery, apneas were induced every time the rat entered REM sleep for 3 h. The same sequence
of apneas was repeated the next day. This way were induced 21 3 apneas/h with average length of 8 s. After apnea termination,
rats usually resumed sleep within 20 s. Apneas longer than 10 s in awake rats induced profound bradycardia (131 14 bpm), a
pressor response (22 3 mmHg), and respiratory effort (pressure swing of > 40 mm Hg). Identical apneas during REM caused a
weak bradycardia (22 3 bpm), and no pressor response (-2 3 mmHg). Respiratory effort also appeared reduced with apneas
during REM
Conclusions:
Responses to apnea in rats are blunted during REM sleep, and resemble those seen in partients with sleep apnea.
Keywords: blood pressure, bradycardia, obstructive apnea, REM, sleep
Resumo:02-104
HYPOXIA INDUCED WITH VASCULAR PROTECTION SOLUTIONS FOR CARDIAC TRANSPLANTATION
INCREASE VASCULAR REACTIVITY.
Simes1; Batista1; Fiorim1; Lima1; Stefanon1; Vassallo1,2

1
Departamento de Cincias Fisiolgicas, UFES
2
Escola Superior de Cincias da Santa Casa de Misericrdia, EMESCAM
Objectives:
Previous studies have demonstrated that the existing methods of myocardial protection are not able to avoid the undesirable
effects of ischemia and the reperfusion phenomenon. However, the mechanisms of action of protecting solutions used for cardiac
transplantation on vascular function are not described yet. This study aims to compare the efficiency of vascular protection by
using the protecting solutions, Krebs-Henseleit (KH), Ringer, Bretschneider-HTK (BHTK), St. Thomas (ST) and Celsior
solutions, in preparation of isolated aorta rings of rats submitted to one hour hypoxia.
Male Wistar rats, 3 months old, were used. The animals were anesthetized and the thoracic aorta was removed and placed in KH
solution for cleaning of connective tissue and fat, and cut in rings with 3 to 5 mm. The record of isometric tension, as percentage
of KCl-induced contraction, of each ring was obtained as proposed by Marin et al. (1998). Vascular reactivity was used to
investigate pressor responses to phenylephrine (PHE, 10-10 to 10-4 M) before and after 1 hour of hypoxia, at 36.5 OC, in rings
exposed to to KH During hypoxia aortic rings were exposed to the protecting solutions, Ringer, BHTK, ST and Celsior, and KH
used as control. Results showed an increased maximum response (Rmax) (KH: 146 14. 5 % n = 7 vs ST: 184 10.5 %, p <
0.05 n = 10) and sensitivity y (pD2) (KH:-6.02 0.5 n = 7 vs ST-8.07 0.24, p < 0.05 n = 8) in aorta rings exposed to ST in KH
solution. There was no change of these parameters with the other protecting solutions.
Conclusions:
This is the first report to show an increased vascular reactivity in aortic rings exposed to the protective solution ST, when
submitted to 1 hour of hypoxia, showing a pattern that suggests endothelial dysfunction. Results suggested that the ST solution
was the less efficient for vascular protection. inducing endothelial injury due to an increased content of ion Ca2 + and K + in its
composition. Results also suggest that the ST solution might be a risk factor for cardiac protection during transplantation
procedures. Grants:CAPES, FAPES/CNPq.
Keywords: BRETSCHNEIDER-HTK, CELSIOR, ISCHEMIA, SAINT THOMAS , VASCULAR PROTECTION
Financial Support: CAPES / CNPq / FAPES- FUNCITEC
Resumo:02-105
LOW CONCENTRATIONS OF LEAD AFTER 7-DAYS OF EXPOSURE INCREASES BLOOD PRESSURE AND
REDUCES THE PARTICIPATION OF NO IN RESISTANCE ARTERIES
Simes, M. R. 1; Furieri, L. B. 1; Fioresi, M. 1,2; Broseghini, G. B. 1; Vescovi, M. V. A. 3; Faria, T. de O. 1;

Martins, L. R. 1; Stefanon, I. 1; Padilha, A. S. 1; Vassallo, D. V. 1,4
1
Depto. Cincias Fisiolgicas/Univ Federal Esprito Santo, UFES
2
Depto. Enfermagem/Univ. Federal Esprito Santo, UFES
3
Depto. Qumica/Univ. Federal Esprito Santo, UFES
4
Escola Superior de Cin. Santa Casa Misericrdia de Vitria, EMESCAM
Objectives:
Evaluate the effects of low concentrations of lead after 7 days of exposure on blood pressure, on the reactivity of mesenteric
arteries of rats and the involvement of NO and the renin-angiotensin system in the vasoconstrictor response.
Male Wistar rats (250-300 g) were divided into two groups: control (Ct) and Lead (Pb). Treated rats received as drinking water
lead acetate (100 ppm) during 7 days and the Ct only water. After this rats were anesthetized and underwent surgery for
catheterization of the carotid artery for measurement of hemodynamic parameters: Systolic Blood Pressure (SBP), Diastolic
Blood Pressure (DBP), Mean Blood Pressure (MBP) and Heart Rate (HR). Blood samples were collected for determination of
lead, using atomic absorption spectrometry and fluorometry for determination of angiotensin converting enzyme (ACE) activity.
After blood collection the animals were sacrificed and the mesenteric arteries (3rd branch) dissected. The mesenteric artery was
divided into segments of 2 mm and mounted in a small vessel four chamber myograph for measurement of isometric tension.
After stabilization, the functional and endothelial integrity were tested. Then, performed concentration-response curves to
phenylephrine (PHE, 10 nM - 30 M). The effects of L-NAME (NOS inhibitor) and enalapril (angiotensin-converting enzyme
inhibitors), on the response to PHE were tested. Statistical analysis: Results were expressed as mean SEM. For comparison of
means Student t test were used. p
Conclusions:
Exposure to low concentrations of lead after 7 days increased blood pressure, did not alter the contractile response to PHE.
However, the exposure was able to reduce the role of NO in resistance arteries from Pb-treated rats. These observations also offer
further evidence that lead short time exposure, even at small concentration, is hazardous and it is an environmental risk factor for
cardiovascular disease
Keywords: BLOOD PRESSURE , RESISTANCE ARTERIES , LEAD
Financial Support: CAPES, CNPq e FAPES
Resumo:02-106
OPIOID RECEPTORS IN THE LATERAL PARABRACHIAL NUCLEUS INVOLVED IN THE CONTROL OF
WATER AND SODIUM INTAKE
Pavan, C. G. ; Penasso, M. G. ; Barbosa, S. P. ; de Luca Jr. , L. A. ; Colombari, D. S. A. ; de Paula, P. M. ;

Colombari, E. ; Menani, J. V.
Depto. de Fisiologia e Patologia, unesp
Objectives:
Bilateral injections of the non specific opioid agonist -endorphin into the lateral parabrachial nucleus increase water and 1.8%
NaCl intake induced by the treatment with the diuretic furosemide (FURO) combined with low doses of the angiotensin
converting enzyme inhibitor captopril (CAP) injected subcutaneously (sc). In the present study, injecting specific agonists and
antagonists into the LPBN, we investigated the participation of and opioid receptors in the LPBN in the control of FURO +
CAP-induced water and 1.8% NaCl intake.
Male Holtzman (290-310 g, n = 8-12) with stainless steel cannulas bilaterally implanted in the LPBN were used. Water and 1.8%
NaCl intake was recorded for 2 to 3 h starting 1 h after the treatment with FURO (10 mg/kg of body weight) + CAP (5 mg/kg of
body weight) sc. Endomorphin-1 ( opioid agonist), CTAP ( opioid antagonist), BLR 52537 ( opioid agonist) or vehicle was
injected into the LPBN 15 to 30 min before starting water and NaCl intake recording. Bilateral injections of endomorphin (1, 2 or
4 nmol/0.2 L) into the LPBN increased FURO + CAP-induced 1.8% NaCl intake (21.1 2.4, 21.4 2.8 and 21.6 3.0 mL/3 h,
respectively, vs. saline: 8.6 1.5 mL/3 h, p
Conclusions:
The results suggest that and opioid receptor activation in the LPBN deactivates the inhibitory mechanisms facilitating NaCl
and water intake induced by FURO + CAP treatment.
Keywords: sodium intake, opioid receptors, lateral parabrachial nucleus
Financial Support: FAPESP, CNPq
Resumo:02-107
ACTIVATION OF POTASSIUM CHANNELS BY NITRIC OXIDE PREVENTS ENDOTHELIAL DYSFUNCTION OF
AORTIC ARTERY IN LEAD-TREATED RATS
Fiorim, J. 1; Jnior, R. F. R. 1; Avevedo, B. F. 1; Padilha, A. S. 1; Stefanon, I. 1; Vassallo, D. V. 1,2

1
Department of Physiological Sciences, UFES
2
Health Science Center of Vitria, EMESCAM
Objectives:
Seven days exposure to low lead concentration increased systolic blood pressure (SBP) and reduced vascular reactivity to
phenylephrine. These results were accompanied by increase in the endothelial modulation and nitric oxide (NO) bioavailability
(Plos One, 6: e17117, 2011). Potassium (K+) channels play a key role in regulating membrane potential and vascular tone.
Activation of K+ channels in vascular smooth muscle leads to hyperpolarization, decrease voltage-gated L-type Ca2+ channel
activity, reduce intracellular [Ca2+], and promote vasodilation. The aim of the present study was to evaluate in aortic rings,
whether 7 days lead-treatment affects both nitric oxide-mediated vasodilator responses and NO dependent activation of K+
channels.
Methods: Wistar rats were treated with lead (1st dose 4 g/100 g, subsequent dose 0.05 g/100 g, i.m. to cover daily loss) or
vehicle. Aortic rings from untreated and lead-treated rats were precontracted with phenylephrine (1 M) or with 60 mmol/l KCl,
and concentration-response curves to acetylcholine (0.1 nM - 300 M) were performed. The participation of NO in the relaxation
induced by ACh was analyzed incubating the vessels with NG-nitro-L-arginine methyl ester (L-NAME, 100 M, nonspecific
NOS inhibitor), for 30 min before phenylephrine or KCl administration. The contribution of K+ channels to ACh-induced
relaxation was assessed in aortas previously incubated for 30-min with K+ channel blockers as follows: tetraethylammonium
(TEA, 2 mM, nonselective blocker of K+ channels), 4-aminopyridine (4-AP, 5 mM, blocker Voltage-dependent K+ channels
KV), charybdotoxin (ChTX, 0.1 M, blocker of Ca2+-activated K+ channels - KCa and KV), iberiotoxin (IbTX, 30 nM, selective
blocker of Large-conductance Ca2+-activated K+ channels - BKCa) and apamin (0.5 M, selective blocker of Small-conductance
Ca2+activated K+ channels - SKCa). In some experiments, concentrationresponse curves to sodium nitroprusside (SNP, 0.01
nM - 0.3 M) were performed. The participation of Kv and BKCa channels in the SNP induced relaxation was analyzed
incubating the rings with 4-AP and IbTX respectively. Results: In control rats, the basal contraction induced by 4-AP was
remarkably greater than that induced by IbTX suggesting a relevant role of KV channels in controlling the aortic vascular tone.
Moreover, basal contractions induced by 4-AP increased after lead-treatment. In phenylephrine precontracted arteries, ACh
induced a similar relaxant response in aortic arteries from both groups that was abolished by L-NAME. However, when arteries
were contracted with high KCl (60 mmol/l) or preincubated with TEA, 4-AP, IbTX, BKCa and SKCa the relaxation elicited by
ACh was reduced more in lead-treated than in control rats. After IbTX and 4-AP preincubation, the relaxant response to SNP was
reduced more in lead-treated than in control rats.
Conclusions:
Our results indicate that 7 days lead-treatment increases activation of K+ channels by NO and this effect might contribute to
preserve the aortic endothelial function.
Keywords: lead, reactivity vascular, nitric oxide , potassium channels
Financial Support: PRONEX- FAPES/CNPq, CAPES
Resumo:02-108
EFFECT OF ACUTE ACHETYLCOLINESTERASE INHIBITION WITH PYRIDOSTIGMINE ON
ELECTROCARDIOGRAM OF ANESTHETIZED RATS.
Santos, F. M. ; Silva Caa ; Salgado, H. C. ; Fazan Jr. , R

Depto. de Fisiologia/Faculdade de Medicina de Ribeirao Preto, FMRP-USP
Objectives:
Several electrocardiographic indices have been proposed to identify patients at risk of sudden death, including the QT interval
length and/or dispersion. We evaluated the potentially benefic effect of an acute increase in parasympathetic drive to the heart,
obtained by pyridostigmine (PYR), an acetyl cholinesterase inhibitor, in anesthetized rats.
After basal recording of electrocardiogram (ECG, lead II), urethane (1 g/kg, ip) anesthetized rats received PYR (0.25 mg/kg) or
isotonic saline intravenously and ECG recording last for 30 min. PYR induced a significant increase in RR (1301 to 1452 ms),
PR (481 to 511 ms) and QT interval (641 to 661 ms). Nevertheless, corrected QT interval (QTc, Bazett formula) decreased
after PYR (1793 to 1742 ms).
Conclusions:
The increase in cholinergic transmission due to the acetycholinesterase inhibition led to an expectable bradycardia with PR
lengthening and a significant shortening in QTc interval. Clinical trials have demonstrated that the prolonged ventricular
repolarization (larger QT interval) is associated with higher incidence of cardiac arrhythmias. The use of PYR as an approach to
enhance vagal drive to the heart might be beneficial in situations of imminent risk of life threatening arrhythmias, such as acute
myocardial infarct.
Keywords: pyridostigmine, electrocardiogram, vagal tone, arrhythmia
Financial Support: FAPESP, CNPq, FAEPA
Resumo:02-109
EFFECTS OF INDAZOL AND RU-INDAZOL IN THE RAT CORPORA CAVERNOSA
Campos, R. M. 1; S, D. S. 2; Sousa, E. S. D. 2; Fonteles, M. C. 1; Lopes, L. G. D. F. 2; Lessa, L. M. A. 1;

Nascimento, N. R. F. D. 1
1
Instituto Superior de Cincias Biomdicas/UECE, ISCB
2
Laboratrio de Bioinorgnica / UFC, UFC
Objectives:
Erectile dysfunction is a highly prevalent disease. One of the most important pathways involved on the mechanism of penile
erection is the increase of intracelular levels of cyclic guanosine monophosphate due to soluble guanylate cyclase (sGC)
activation. The aim of this present study is to investigate the effects of a metal-based drug, derived from the soluble guanylate
cyclase activator 3-(5-hydroxymethyl-2-furyl)-1-benzyl indazole (YC-1), on the relaxation of corpora cavernosa smooth muscle.
The pharmaceutical industry is searching a sGC- stimulating compound that is neither dependent on nitric oxide (NO)
release/activity nor on the redox status of the haem-group of the enzyme. The potential impact of such drugs relies in the fact that
they are able to increase the rate of activity of oxidized sGC a phenotype that is prevalent under disease conditions, especially
where increased oxidative stress is present, such as diabetes. Our group has synthesized a rutheniumbased compound named
Ruindazol that is probed here for corpora cavernosa relaxation compared to the lead compound, indazol
Male Wistar rats (250g-300g) were sacrificed with sodium thiopental (200 mg/Kg;,i.p). Thereafter, the penis was excised out and
careful dissecation was performed in order to remove surrounding tissues. Strips of corpora cavernosa were immersed in KrebsHenseleit solution kept at 37C and gassed continuously with 95% O2 in 5% CO2. The tissues were fixed in an isometric force
transducer under 0.5g resting tension and kept at equilibration conditions for 60 minutes-period. Modifications of resting tension
were continuously recorded in a 4-channel physiograph ( Narco Biosystems , Houston, Texas, EUA). Tissues were pre-contracted
with phenylephrine (PE; 20 M) and then concentration-response curves to Indazol (1nM - 100M) and its derivative compound,
RU-Indazol (1nM - 100M) were performed. In another set of experiments, the concentration- response curves to both
compounds were repeated in strips treated with a soluble guanylate cyclase inhibitor [1,2,4]oxalodiazolo[4,3]oxonalina (ODQ;
100 M). These protocols were approved by the local Ethicall Commitee for Animal Experimentation (protocol number:
08628332-4). The data were analyzed by Students t test ( p<0.05).
Conclusions:
RU-Indazol induced a concentration-dependent relaxation of strips of corpora cavernosa with a maximal relaxant response of
100% and IC50 of 8.9 M (n=5). Indazol promoted a maximal response of 67.6% 8.0 (p
Keywords: endothelial dysfunction, nitrergic dysfunction, oxidized soluble guanylate cyclase, soluble guanylate cyclase
activators
Financial Support: This study was supported by CNPq/ Brazil
Resumo:02-110
ROLE OF REDOX-SENSITIVE PROTEINS IN THE ESTABLISHMENT OF CARDIAC HYPERTROPHY
MEDIATED BY RENIN-ANGIOTENSIN SYSTEM IN HYPERTHYROIDISM.
Baraldi, D. D. ; Berger, B. R. ; Conzatti, A. ; Schenckel, P. C. ; Fernandes, T. R. G. ; Carraro, C. C. ; Bellklein, A. ; Arajo, A. S. R.

Fisiologia, UFRGS
Objectives:
Hyperthyroidism generates important effects on the basal metabolism and cardiovascular system, altering its function and leading
to cardiac hypertrophy. It has been reported the relevant role of renin-angiotensin system (RAS) in the development of cardiac
hypertrophy in hyperthyroidism. In this context, angiotensin II would bind to the AT1 receptor, leading to the activation of
gp91phox (a NADPH oxidase subunit). This enzyme is involved in the reactive oxygen species (ROS) production, such as
superoxide anion and hydrogen peroxide (H2O2). ROS may activate the transcription factor (NrF2), which regulate protein
expression of antioxidants, such as heme-oxygenase-1 (HO-1). Therefore, the aim of this study was to investigate association
amongst H2O2, NrF2, HO-1, and renin-angiotensin system in the establishment of thyroid hormone-induced cardiac hypertrophy.
Male Wistar rats (n=5/group) were divided in: control (C), losartan (L) (10mg/Kg/day, intragastric, 28 days), L-thyroxin (T4)
(12mg/L in drinking water, 28 days), and T4+losartan (T4+L). The animals were catheterized to measure the resting heart rate
(HR). After the treatment period, animals were decapitated and its hearts were excised to perform: cardiac hypertrophy index
(CHI) (heart weight/body weight), and H2O2 concentration (nmol/g tissue). The protein expression of AT1 receptor, gp91phox,
NrF-2 and HO was evaluated by Western Blot. To compare groups, two way ANOVA with Student-Newmann-Keuls post hoc
test was used. Differences were considered to be statistically significant at p
Conclusions:
The findings of this study showed that renin-angiotensin system is associated with the development of cardiac hypertrophy
induced by thyroid hormones, being this process possibly modulated by reactive oxygen species. The RAS blockade was able to
reduce hypertrophy development, with the simultaneous reduction in the expression of proteins involved with the cell redox
regulation. These data suggest a connection among cardiac RAS, cellular redox status and its regulators proteins in the
establishment of cardiac hypertrophy in hyperthyroidism.
Keywords: hyperthyroidism, oxidative stress, renin-angiotensin system, cardiac hypetrophy, cell signalling
Financial Support: CNPq, CAPES e FAPERGS.
Resumo:02-111
TOLL-LIKE RECEPTOR 2 IS INCREASED IN RENAL ISCHEMIA/REPERFUSION-INDUCED CARDIAC
HYPERTROPHY IN MICE
Trentin-sonoda M ; Cirino-silva R. ; Carneiro-ramos Ms

Universidade Federal do ABC, UFABC
Objectives:
The innate immune response can exert innumerous effects in vast number of tissues and organs. Its well established that
cytokines and Toll-like receptors (TLRs) participate of cardiac trophism modulation. The aim of this study was to investigate the
role of TLRs in heart after renal ischemia/reperfusion.
C57bl/6J mice were submitted to unilateral 60 minutes kidney ischemia, followed by reperfusion for 5, 8, 15 and 20 days (n=6
per group). Levels of urea were measured to confirm renal failure as well vimentin mRNA expression. Cardiac ANF gene
expression was used as molecular cardiac hypertrophy marker and all gene expression were analyzed by real time PCR. Urea
levels and mRNA Vimentin were increased in groups 5, 15 and 20 days (P
Conclusions:
First, the results indicate that renal ischemia/reperfusion model is able to induce a transient cardiac hypertrophy in mice.
Moreover, this hypertrophy has a positive correlation with a transient increase of TLR2 mRNA levels, suggesting the possible
involvement of TLR2 cascade activation in cardiac hypertrophy development model. Is well known that TLRs play an important
role in cardiac hypertrophy models, probably through NF-kB or Akt/mTOR activation signaling. Herein, we showed an
experimental model that TLR2 can participate of cardiac hypertrophy process.
Keywords: Renal ischemia/reperfusion, Cardiac hypertrophy, Toll-like receptors
Resumo:02-112
VENODILATATION INDUCED BY TERPY INVOLVES ACTIVATION OF SGC, K+ CHANNELS KV, SKCA AND
SARCOPLASMIC RETICULUM CA-ATPASE
Paulo, M. 1; Grando, M. D. 1; Rodrigues, G. J. 2; Dasilva, R. S. 1; Bendhack, L. M. 1

1
Physics and Chemistry, Faculty of Pharmaceutical Sciences , FCFRP-USP
2
Pharmacology,School of Medicine of Ribeiro Preto, FMPR-USP
Objectives:
Nitric oxide (NO) donors have important therapeutic effect for treatment of cardiovascular diseases such as angina pectoris and
hypertension. A major clinical benefit of NO donors is attributed to their venodilator effect, resulting in decreased venous return,
cardiac preload, blood pressure and myocardial oxygen demand. But the most common side effect of these drugs is headache,
which is caused by cerebral vasodilatation. Aim: This study aimed to verify the vasorelaxation induced by Terpy, in basilar artery
and cava vein from 2K (normotensive) and 2K-1C (hypertensive) rats. It was also evaluated the cellular mechanisms involved in
this relaxation.
The NO donor Terpy was synthesized in our laboratory. We have analyzed the maximal relaxation (Emax) and potency (pEC50)
of Terpy in cava vein and basilar artery pre-contracted with endothelin-1 (ET-1) after incubation with ODQ (1M), guanylylcyclase inhibitor (sGC); thapsigargin (1M) sarcoplasmic reticulum Ca2+-ATPase (SERCA) inhibitor; Rp-8-Br-PET-cGMP 3
M G-kinase (GK) inhibitor; dipiridamole (3 M) a phosphodiesterase-5 (PDE5) inhibitor; TEA (1mM) a non-selective K+
blocker and selective K+ blockers: apamin (small-conductance Ca2+-activated-SKca), 4-aminopyridine (voltage-dependent Kv)
and glibenclamide (ATP-sensitive KATP). We evaluated the NO released from Terpy by using confocal microscopy. Vascular
reactivity experiments showed that Terpy did not induce relaxation of the basilar artery from 2K and 2K-1C rats.Terpy did not
release NO in basilar artery. In cava vein, Terpy released NO and induced vascular relaxation. pEC50 and the Emax of Terpy
were lower in 2K-1C (pEC50: 5.42 0.22; Emax: 56.7 3.1%, n=7) than 2K vein ( pEC50: 6.10 0.12; Emax: 70.3 3.2% n=6,
p
Conclusions:
Taken together, our results demonstrate that Terpy does not release NO and it does not induce relaxation in basilar artery from 2K
and 2K1C rats. Terpy releases NO in cava vein and it induces relaxation in both 2K and 2K-1C rat veins, which was lower in 2K-
1C than in 2K. The venodilatation induced by Terpy involves activation of sGC, K+ channels Kv and SKCa and SERCA.
Keywords: Hypertension, Nitric oxide donor, Potassium channels, SERCA, Venodilatation
Financial Support: FAPESP and CNPq
Resumo:02-113
TREATMENT FOR 15 DAYS WITH TRIBUTYLTIN DECREASE THE VASCULAR REACTIVITY TO
PHENYLEPRINE IN ISOLATE AORTA RINGS OF FEMALE RATS
Rodrigues, S. M. L. ; Ximenes, C. F. ; Batista, P. R. D. ; Simes, F. V. ; Filho, V. S. D. ; Graceli, J. B. ;

Vassalo, D. V. ; Stefanon, I.
Departamento de cincias fisiolgicas, UFES
Objectives:
Organotin compounds such as tributyltin (TBT) are used as antifouling paints for shipping companies and as pesticides in
agriculture. It can be trasnsmitted to aquatic organisms and humans by biotransference through the food chain. TBT may cause
endocrine dysfunction by the influence on female gonadal hormones inhibing the aromatase responsible to transforms
testosterone in estrogen. Our hypothesis is that TBT, by inhibiting the estrogen synthesis, could prejudice its cardiovascular
protection. This study aimed to investigate the effects of 15 days exposure to TBT (20 ng/ml, oral administration) on vascular
reactivity to phenylephrine (PHE) in isolated aorta rings of female rats.
Female Wistar rats (230-250 g) were divided in control group, treated with the vehicle (CT, n= 8) and the TBT group, treated
with TBT solution (20ng/ml) (TBT, n= 8) by gavage for 15 consecutive days. All the protocols were developed was approved by
the UFES Animals Ethical Committee (n. 020/2009; vascular reactivity n. 004/2007). In the end of treatment all animals were
anesthetized with pentobarbital (35mg/kg, i.p.) and the blood was collected for serum levels of 17 -estradiol (E2) and
progesterone (P4). The thoracic aorta was removed and cutted in rings, immersed in 5 ml of Krebs solution at 37 C, pH 7.4. The
rings were submitted the rings to crescent concentration of phenyleprine (10-10 10-4 M). The same curve was held in the
absence of endothelium, in the presence of L-NAME (NOS blocker), tetraetilamnio-TEA (K+ channel blocker) and apocinina
(NADPH inhibitor). The data were expressed as mean SEM and analysed by Student's t test non-paired, significant for p < 0.05.
The treatment with TBT decreased levels of E2 (CT:47.27.1 vs TBT:32.24.3pg/mL, p< 0.05). TBT decreased the maximal
response to PHE (CT: 145.7 vs TBT: 122.4 7.6, p < 0.05). In the abscense of endothelium the TBT group presented a greater
Rmx than control (CT: 152.6 8.3 vs TBT: 191.1 26.5, p < 0.05). The L-NAME and TEA increment of reactivity was greater
in the TBT group (L-NAME/CT: 149.1 9 vs. TBT: 157.8 3.3, p < 0.05; TEA/CT: 158.1 10.2 vs TBT: 162.5 6.7, p <
0.05). The decrease in the reactivity to PHE during incubation with apocinina was more evident in the TBT group (TC: 70.1
10.1 vs TBT: 59.13 8.9, p < 0.05).
Conclusions:
The treatment of female rats with TBT for 15 days decreased the vascular reactivity to PHE probably for increase dependent on
the NO participation and the K+ channels.
Keywords: Female gonadal hormones, Nitric oxide synthase(NOS), Oxidative stress, Tributyltin, Vascular reactitivity
Financial Support: CAPES / CNPq / FAPES-FACTEC
Resumo:02-114
AUTONOMIC CHANGES AFTER RESISTANCE EXERCISE IN INDIVIDUALS WITH UNTREATED STAGE 1
HYPERTENSION.
Chrispino, T. C. ; Barbosa, T. P. C. ; Neves, F. J. ; Nbrega, A. C. L.

Fisiologia e Farmacologia, UFF
Objectives:
Subjects with hypertension have impaired autonomic modulation, whereas aerobic exercise training improves this profile.
However, there is no clear evidence of autonomic modulation in response to resistance exercise. Our aim was to evaluate the
autonomic response after one bout of resistance exercise in patients with untreated stage 1 hypertension.
Six men with stage 1 hypertension (age 428 yrs; systolic blood pressure between 140 and 159 mmHg and/or diastolic blood
pressure between 90 and 99 mmHg) underwent a randomized and crossover study. All of the subjects fulfilled the following
criteria: age between 20 and 60 years, body mass index lower than 35 kg/m, not using medications, absence of any diagnosed
chronic disease, and sedentary. Subjects underwent medical examination and laboratory tests, one repetition maximum test.
Autonomic modulation was assessed by analysis of signals from a digital infrared photoplethysmographer (Finometer) yielding
both time and frequency domain indices of heart rate variability and arterial baroreflex sensitivity by the alpha index. The
experimental protocol included two days: one with (intervention day) the other and without (control day) one 45-min session of
resistance exercise of 6 body movements composed of 3 series of 20 repetitions at 40% 1RM. Ambulatory blood pressure
monitoring was applied on both days. The protocol was approved by the Ethics Committee (CCM/HUAP 180/08). Systolic blood
pressure was lower after exercise (baseline: 1456; 0h: 1303 mmHg, p0.05).
Conclusions:
These findings suggest that individuals with stage 1 hypertension have lower parasympathetic modulation and baroreflex
sensitivity immediately after completion of resistance exercise. The reduced baroreflex sensitivity may be involved in the
mechanisms responsible for post-resistance exercise hypotension.
Keywords: Autonomic Nervous System, Hypertension, Resistance Exercise
Financial Support: Capes, CNPq, FAPERJ, FINEP, Labs D'Or.
Resumo:03-065
EFFECT OF SWIMMING TRAINING ON PLATELET AGGREGATION IN HYPERTENSIVE RATS
Cardoso, A. M. 1; Fiorin, F. S. 1; Dutra, E. J. M. 1; Martins, C. C. 1; Abdalla, F. H. 1; Pereira, L. B. 1;
Bagatini, M. D. 2; Stefanello, N. 1; Ferreira, R. 1; Schetinger, M. R. C. 1

1
Depto de Qumica/ Universidade Federal de Santa Maria, UFSM
2
Universidade Federal da Fronteira Sul , UFFS
Objectives:
Currently, hypertension affects more than 1 billion adults worldwide and 9095% of these patients have essential hypertension. It
is well known that in animal models the inhibition of nitric oxide biosynthesis by administration of N-Nitro-L-arginine methyl
ester hydrochloride (L-NAME) leads to arterial hypertension. Spontaneous platelet aggregation is largely reported in
hypertension cases and its specific role in the pathophysiology of this disease must be considered since platelets can promote
thrombotic complications. In the other hand exercise training has been associated with several metabolic and cardiovascular
benefits and has been recommended for prevention as well as for treatment of cardiovascular diseases. Thus, the aim of this work
was to investigate the effect of swimming training on platelet aggregation in adult male rats with hypertension induced by LNAME administration.
The rats were maintained on a 12:12h light dark photoperiod and fed ad libtum. They were randomly divided in four groups. LNAME (n= 10): Rats were administered nitro-L-arginine methyl ester (30mg/kg/day) orally, by gavage, in the morning for 8
weeks and put on water for 5 min. Exercise L-NAME (n=10): was administered the same L-NAME dose as the first group and
the animals were submitted to swimming effects, from monday to friday, 1h/day, with gradual increase in workload up to it
reaches 5% of body weight in a adapted swimming system for rats with heated water to 31 1C. Control (n=10): normotensive
and sedentary animals (put on water for 5 min). Exercise Control (n=10): normotensive animals submitted to an exercise effects.
The animals were euthanized and blood was withdrawn and used for the separation of platelet rich plasma. For platelet
aggregation, the Born (J. Physiol. 95: 168, 1963) technique was performed by turbidimetric measurement with a Chrono-log
optical aggregometer, with AGGRO/LINK Model 810-CA software for Windows version 5.1. Aggregation was measured at
37C and expressed as the maximal percent change in light transmittance from baseline at 5 min after the addition of the agonist
adenosine diphosphate (ADP) 10 M, with platelet poor plasma as a reference. Data were analyzed using 2-way ANOVA,
considering p
Conclusions:
In this study, we observed that platelet aggregation is increased in platelets from L-NAME-treated rats, which can be a risk factor
for atherosclerosis development and thrombotic complications. However, we showed that swimming training has the ability to
modulate platelet aggregation, and, this way, can act as inhibitor of thrombus formation, possessing cardio protector effects that
could be considered as a non-pharmacological treatment for hypertension.
Keywords: Hypertension, Exercise training , L-NAME , Platelet aggregation
Financial Support: Sources of research support: CNPq
Resumo:03-066
INHIBITION OF BRAIN NITRIC OXIDE SYNTHESIS DECREASES RUNNING PERFORMANCE BY
ATTENUATING CUTANEOUS HEAT LOSS MECHANISMS
Lima, P. M. A. ; Coimbra, C. C.
Department of Physiology and Biophysics, ICB / UFMG
Objectives:
To asses the influence of central inhibition of nitric oxide synthesis on physical permorfance and thermoregulatory adjustments in
untrained rats during submaximal-intensity exercise on a treadmill until fatigue.
Either 2 L of 1.43 mol N-nitro-L-arginine methyl ester (L-NAME, n = 7) or 0.15 M NaCl solution (SAL, n = 7) were injected
into the right lateral cerebral ventricle of male Wistar rats (250-350 g) immediately before the animal was subjected to running
exercise (18 m.min-1, 5% inclination until fatigue). During exercise, tail temperature (Tt) was measured using a thermocouple
taped to the tail, and intraperitoneal temperature, an index of core temperature (Tc), was recorded using a telemetry sensor
implanted into the peritoneal cavity. Heat storage rate (HSR) was calculated. Rats injected with L-NAME showed a 43%
reduction in running performance (33.0 4.5 min L-NAME vs. 57.7 10.8 min SAL, p<0.05).
Conclusions:
Our data showed that central inhibition of the brain nitrergic system decreased the ability to dissipate heat, leading to decreased
physical performance during a submaximal-intensity exercise on the treadmill. We concluded that central nitric oxide
transmission modulates cutaneous heat dissipation.
Keywords: Exercise, Heat storage rate, Heat dissipation, Nitric oxide
Financial Support: CNPq, Capes and Fapemig
Resumo:03-067
LEUCINE SUPPLEMENTATION ENHANCES THE ADAPTATIONS OF ENDURANCE TRAINING ON SKELETAL
MUSCLE IN A GENETIC MODEL OF SYMPATHETIC HYPERACTIVITY-INDUCED HEART FAILURE
Moraes, W. M. A. M. D. 1; Souza, P. R. M. D. 3; Guimares, F. D. 1; B. Jr. , C. R. 4; Zatz, M. 4; Negro, C.

E. 1; Brum, P. C. 1; Medeiros, A. 2
1
School of Physical Education and Sport - USP, EEFE - USP
2
Department of Biosciences - UNIFESP , UNIFESP
3
Experimental Hypertension Laboratory , Heart Institute -USP
4
Human Genome Research Center , HGRC- USP
Objectives:
To investigate the effects of leucine supplementation associated with endurance training (ET) on function and morphology of
skeletal muscle in a genetic model of sympathetic hyperactivity-induced heart failure in mice, and whether these effects were
associated with activation of mTOR pathway.
Treatment consisted of 4-wk of ET on a motor treadmill, wich consisted in sessions of 60 min based on maximum lactate steady
state (6d/wk), and administration of leucine (1.35g/kg) or placebo (distilled water) intragastrically. We established five groups: a
control without heart failure (WT, n=12 and four groups of mice lacking both 2A and 2C adrenergic receptor subtypes (KO),
which were randomly divided into sedentary receiving placebo (KO, n=12) or leucine (KOL, n=12); trained receiving placebo
(KOT, n=12) or trained receiving leucine (KOLT, n=12). It was analyzed exercise capacity by graded treadmill exercise protocol
performed until exhaustion, cross-sectional area (CSA) by histochemical myosin ATPase, muscle function by ambulation and
grip tests, protein expression levels by western blotting, and mortality curve (logHank). KOT displayed increased exercise
capacity by 14% compared to test performed in the pre-experimental period, increased CSA in type I fibers in soleus muscle (10
and 11% compared to KO and KOL, respectively) and type IIA and IIB in plantaris muscle (13 and 15% compared to KO and
KOL, respectively), and improved performance in ambulation (13%) and grip tests (11%) vs KO and KOL. The leucine
supplementation alone showed no effect on muscle function or the phenotype of the fibers, but associated to ET improved CSA in
IIA and IIB fibers in plantaris muscle, and muscle function, at a rate greater than improve in KOT. The leucine supplementation
associated to ET was the only intervention able to improve survival. Additionally we found that these effects observed in KOLT
were associated with increased protein expression levels of phospho-mTOR:mTOR, phospho-p70S6K:p70S6K and a reduction of
phospho-AMPK:AMPK in 34, 47 and 30% compared to KOT, respectively.
Conclusions:
Supplementation of leucine enhances the effects of ET to improve exercise capacity, to preserve the mass and prevent muscle
disorders. These effects are specific to the glycolytic fibers and are associated with the activation of mTOR pathway and
improved survival.
Keywords: endurance training, leucine, heart failure, skeletal muscle
Resumo:03-068
LACK OF BETA2-ADRENOCEPTORS ACELLERATES SKELETAL MUSCLE ATROPHY IN ISCHEMIAINDUCED HEART FAILURE MICE.
Voltarelli, V. A. ; Bechara, L. R. G. ; Mattos, K. C. ; Bacurau, A. V. N. ; Brum, P. C.

Escola de Educao Fsica e Esporte da USP, EEFE-USP
Objectives:
Sympathetic hyperactivity is a hallmark of heart failure (HF) and it exerts a toxic effect on cardiac muscle. However, less is
known about the effects of sympathetic hyperactivity on skeletal muscle (SM) in HF. As 2-adrenoceptors mediate the effect of
sympathetic activity in SM being targets of circulating catecholamines, we evaluated SM structure and phenotype of mice with
target disruption of 2-adrenoceptor gene (2KO) after ischemia-induced HF. Furthermore, we analyzed the expression of key
proteins involved in 2-adrenoceptor signaling related to SM hypertrophy and atrophy.
A cohort of male 2KO (KO) mice in a FVB genetic background and their wild-type (WT) controls were submitted to myocardial
infarction (MI) or SHAM surgery at 4 mo of age. After 30 days of surgeries, fractional shortening (FS) was determined by
echocardiography. Exercise tolerance was estimated at 30, 45, 75 and 90 days post-infarction using a graded treadmill exercise
test. Animals were killed at 90 days post-surgeries. Fiber typing, cross-sectional area (CSA) and capillary density were evaluated
by mATPase histochemistry in plantaris and soleus muscles. SM protein expression of Akt1, p-Akt1, Foxo3a, p-Foxo3a, MuRF1
and Atrogin1 was determined by Western Blotting in plantaris muscle. The animals were divided into four groups: CO SHAM,
CO MI, KO SHAM and KO MI. With a mortality rate of 56%, the MI was effective in reducing cardiac function (FS: 23,0
2,3% CO MI vs. 39,0 1,2% CO SHAM and 24,0 3,5% KO MI vs. 38,0 0,6% KO SHAM) and caused hypertrophy and
dilation of left ventricular in infarcted mice. Nineteendays post-surgery, exercise tolerance was significantly reduced in both MI
groups when compared with SHAM groups and this decrease was greater in KO mice (-44% KO MI vs. -32% CO MI). Likewise,
both CO MI and KO MI showed capillary rarefaction associated with decreased type I distribution and increase of glycolytic
fibers in plantaris and soleous. Nevertheless, only KO MI animals displayed reduced CSA of fiber types I, IIA, and IIB in
plantaris and I and IIA in soleous. Among the proteins that were quantified, MI increased the expression of Atrogin1 in both CO
and KO groups when compared to their respective control groups (142% of control for CO MI and 146% of control for KO MI),
but only KO MI mice presented increased expression of MuRF1 in plantaris (210% of control). There were no statistical
differences for Akt1, p-Akt1, Foxo3a and p-Foxo3a proteins between groups.
Conclusions:
Taken together, these results suggest that lack of 2-adrenoceptors accelerates skeletal muscle atrophy in HF, at least at the stage
of HF presently studied, and proteasome ubiquitin system seems to be associated with this finding.
Keywords: Heart failure, Skeletal muscle, Beta2-adrenoceptors
Resumo:03-069
AEROBIC EXERCISE TRAINING IMPROVES CARDIAC MITOCHONDRIAL FUNCTION AND REDOX
BALANCE STATUS IN RATS
Campos, J. C. 1; Queliconi, B. B. 2; Gomes, K. M. S. 1; Dourado, P. M. M. 3; Brum, P. C. 1; Kowaltowski,

A. J. 2; Ferreira, J. C. B. 1
1
Escola de Educao Fsica e Esporte, EEFE-USP
2
Instituto de Qumica, IQ-USP
3
Instituto do Corao, InCor-HCFMUSP
Objectives:
Over the last decades, exercise training has been widely recognized as an important and efficient strategy for improving quality of
life. Indeed, exercise training is also an important therapy for preventing and treating a variety of diseases. Resting bradycardia
and increased exercise tolerance are well-known markers of exercise training effectiveness. However, the underlying cellular
mechanisms by which exercise training improves aerobic capacity are not completely understood. In the present study, we aimed
to investigate the effect of eight weeks of aerobic exercise training on cardiac mitochondrial function and redox balance in rats.
Male Wistar rats (3 months of age) were randomly assigned into sedentary and trained groups. All rats performed a graded
treadmill exercise test until exhaustion before and after exercise training protocol. Aerobic exercise training consisted of 8 weeks
of running on a motor treadmill, 5 days/week, for 60 min at 60% of the maximal speed achieved in the graded treadmill exercise
test. The animal care and protocols were reviewed and approved by the Ethical Committee of Medical School of University of
So Paulo (2009/56). Trained rats displayed resting bradicardia (from 48517 before, to 40416 bpm after training protocol) an
improved running performance (from 45148 before, to 83559 meters after training protocol) when compared to sedentary
animals. Eight weeks of exercise training improved mitochondrial function, found by increased state 3 (10012 vs. 20558%)
and state 4 (10013 vs. 16613%) respiratory rates compared to sedentary animals. Indeed, mitochondria isolated from trained
hearts displayed increased ROS production under either complex I (10011 vs. 26062%) or complex III (1006 vs. 24969%)
inhibition by rotenone and antimycin A, respectively. Of interest, exercise training reduced ROS production in state 3 (10015
vs. 6414%) and state 4 (10013 vs. 5812%) respiratory rates when corrected by the total oxygen uptake. Furthermore, trained
rats increased calcium uptake (1006 vs. 16025%) and decreased inner membrane potential (1001 vs. 844%) when compared
with sedentary group.
Conclusions:
The recognized effects of aerobic exercise training such as, like enhanced aerobic capacity and decreased resting heart rate, was
associated with improves in cardiac mitochondrial function, presenting increased state 3 and state 4 respiratory rates, maximal
calcium uptake and decreased ROS production in cardiac isolated mitochondria. Our findings support the hypothesis that
enhanced mitochondrial function by increasing respiratory control ratio, improving maximal calcium uptake and reducing ROS
production underlies, at least in part, the positive effects of aerobic exercise training. These findings reinforce the contribution of
exercise training as a non-pharmacological therapy for prevention and treatment of mitochondrial dysfunction-associated
diseases.
Keywords: Exercise training, Cardiac mitochondrial function, Redox Balance
Financial Support: This study was supported by FAPESP (#2009/12349-2, #2010/00028-4).
Resumo:03-070
FUNCTIONAL ASSESSMENT OF INDOOR UNDER-17 SOCCER ATHLETES BELONGING TO THE CITY TEAM
Romero, P. V. S. ; Peserico, C. S. ; Mezzaroba, P. V. ; Esteves, J. V. D. C. ; Andreato, L. V. ; Moraes, S. M.

F. ; Machado, F. A.
Cincias Fisiolgicas / Universidade Estadual de Maring, UEM
Objectives:
To evaluate and quantify the physical fitness along with its motor behavior of indoor soccer athletes from the sub-17 category.
The sample was composed of 14 indoor soccer athletes from the sub-17 category, 3 of them being goalkeepers, 2 fixed, 6 wings
and 3 pivots. It was used anthropometric measures of body mass, height and skin folds. To verify the body weight it was used a
Tanita Digital scale to measure the height, we used the stadiometer. We calculated the body density using Jackson and Pollock
(1978) and for the determination of the fat % it was calculated using the Siri formula (1961). We evaluated parameters related to
the flexibility in sitting and to reach over Wells and Dillon (POLLOCK; WILMORE, 1993), abdominal strength in 1 minute
(AAHPER, 1976), power by the test of grip strength (JOHNSON, NELSON, 1979) and muscle strength in the horizontal
impulsion (JOHNSON;NELSON, 1979), in addition to the speeding tests of 30 meters (POPOV, 1986) and agility of SEMO
(JOHNSON, NELSON, 1979). Tests were carried out to race on a treadmill (Inbrasport, Porto Alegre, RS, Brazil) in airconditioned laboratory, with temperature maintained between 20 C and 24 C. The initial velocity was 7 km/h with increments
of 1 km/h every 3 minutes, and it kept and constant 1% inclination. The athletes were strongly encouraged to remain in the test
until the voluntary exhaustion. Ventilatory variables were determined by ergoespirometry, it was used a gas analyzer (Spirometer
VO2000, Inbrasport, Porto Alegre, Brazil), in addition to an electrocardiograph to record the heart rate through the program
ERGO PC (Micromed, Brasilia - Brazil). This study was approved by the Standing Committee on Ethics in Research Involving
Human Beings (COPEP), of the State University of Maring, under the opinion no. 175/2007. The results were compatible with
the physical tasks required to this sport practice. The athletes were aged (years) 16.79 0.97, height (cm) 175.22 5.18 , BMI
(Kg/m2) 21.56 1.46 . The anthropometric measurements and body composition indicated a low fat % (7.99 3.32 ). The
flexibility of 32.96 6.32 cm was classified as averaged, even though it is not the ideal physical condition required by the sport.
The results obtained for determining the motor suitability, presented: abdominal Strength 45.14 10.45 , lower limbs
strength101.35 40.13 , Speed of 30 meters (s) 4.77 0.23 , Agility SEMO (s) 11.74 0.53 , Horizontal Jump (cm) 220.75
15.35 . The results that showed the cardiorespiratory fitness presented the average values of 59.68 6.71 VO2max (ml/kg min),
193.1 6.08 hrmax (bpm) and Vmax (km/h) 15.40 1.17. These values are presented in avarage standard deviation (SD). The
normality of the data was confirmed by the Shapiro-wilk test using the SPSS 13.0.
Conclusions:
The maringaenses athletes of the sub-17 futsal category showed a morphological profile which is compatible with data of athletes
from other studies involving teams of high-performance, making them suitable to carry out the work-tactical-physical technique
required for sports practice.
Keywords: Futsal, Physical fitness, Motor behavior
Financial Support: CNPq / FADEC.
Resumo:03-071
EFFECT OF THE AEROBIC PHYSICAL CONDITIONING LEVEL ON CARDIAC AUTONOMIC MODULATION
IN HEALTHY SUBJECTS
Dutra, S. G. V. ; Pereira, A. P. M. ; Mazon, J. H. ; Maida, K. D. ; Tezini, G. C. S. V. ; Souza, H. C. D.

Depto Biomecnica e Reabilitao / Universidade de So Paulo, FMRP/USP
Objectives:
The practice of physical exercises periodically promotes beneficial adaptations in the cardiovascular autonomic control, being
largely used for prevention and in association to applied therapy for control of chronic-degenerative diseases (Eur. J. Cardiovasc.
Prev. Rehabil. 14:237, 2007). However, although these adaptations have been widely investigated in several physiopathological
situations, studies are scanty on healthy subjects, mainly regarding the effect of physical conditioning level on cardiac autonomic
control. In this way, the objective of the present study was to investigate the relationship between level of physical conditioning
and cardiac autonomic modulation in healthy subjects.
Fifty-three healthy men (aged between 18-45 years old) were submitted to maximum ergoespirometric test (Ellestad) and then
divided into three groups: Sedentary (VO2peak = 22-38 ml.kg-1.min-1, SED, n = 18); Moderate Performance (VO2peak = 38-48
ml.kg-1.min-1, MP, n = 22); and High Performance (VO2peak > 48 ml.kg-1.min-1, HP, n = 13). Analysis of the heart rate variability
(HRV) was performed by using the autoregressive method at R-R intervals of electrocardiographic record (ECG) in three
moments during ergospirometric test: at rest (baseline), 3 (Rec3) and 6 minutes (Rec6) following the beginning of recovery.
Peripheral blood collection for analysis of lactate was performed at rest, during the peak, and 6 minutes after recovery (Rec6).
Data were presented as mean standard deviation and Kruskal-Wallis test was used for statistical test (p < 0.05). This study was
approved by the local ethics research committee under the protocol number 8381/2010. The levels of lactate at baseline and peak
moments were lower in group SED (1.90.5 and 6.41.0 mmol.L -1, respectively) when compared to the other groups (MP:
2.40.8 and 7.00.9 mmol.L-1; HP: 2.50.4 and 7.51.2 mmol.L-1). Nevertheless, the latter group (3.90.9 mmol.L-1) showed
lower levels of lactate at Rec6 compared to group MP (5.81.9 mmol.L -1), and this in relation to group SED (7.41.1mmol.L-1).
For HRV at rest, no differences were found in any of the spectral parameters. At Rec3, groups MP (Variance: 2.42.3ms 2; LF:
1.21.2ms2; HF: 0.40.5ms2) and HP (Variance: 1.91.9ms2; LF: 1.21.5ms2; HF: 0.60.9ms2) showed increases in variance as
well as in LF and HF oscillations compared to group SED (Variance: 0.70.7ms 2; LF: 0.30.5ms2; HF: 0.10.2ms2). At Rec6,
group HP (Variance: 4.33.5ms2; LF: 2.92.6ms2; HF: 1.82.6ms2) showed increases in variance as well as in LF and HF
oscillations compared to groups SED (Variance: 1.21.4ms 2; LF: 0.91.1ms2; HF: 0.20.1ms2) and MP (Variance: 2.83.1ms2;
LF: 2.42.6ms2; HF: 0.60.8ms2).

Conclusions:
The level of aerobic physical conditioning does not interfere with cardiac autonomic modulation at rest, but physically
conditioned subjects seem to present a better capacity to resume the baseline values for cardiac autonomic modulation.
Keywords: cardiac autonomic modulation, heart rate variability, physical conditioning, physical exercise, lactate
Financial Support: Source of research support: CAPES.
Resumo:03-072
LEVELS OF CIRCULATING PROGENITOR CELLS AND VASCULAR FUNCTION IN ENDURANCE ATHLETES.
Machado, R. C. ; Baldo, M. P. ; Roelke, L. H. ; Forechi, L. ; Verbeno, B. ; Vassallo, P. F. ; Mill, J. G.

DEPARTAMENTO DE CINCIAS FISIOLGICAS, UFES
Objectives:
Recent evidences have shown that vascular function depends not only on resident cells in blood vessel wall but also seems to be
modulated by circulating progenitor cells CPCs derived from bone marrow. Physical training induces mobilization of either
CPCs and the subpopulation of endothelial progenitor cells EPCs. Moreover, cell mobilization seems to improve the endothelial
function in presence of established vascular disease and or cardiovascular risk factors. However, such effects in healthy subjects
are still poorly studied. Therefore, our aim was investigate the influence of longterm endurance exercise training on the different
subpopulations of CPC CD34+, CD133+, CD34+CD133+ and CD34+KDR+ EPC in amateur runners and healthy sedentary
subjects at rest, and evaluate the artery endothelial function by brachial flowmediated dilatation FMD in this subjects.
The study population was consisted of participants from the Cardiovascular Health Study of Trained Man ESCHOT.
Longterm runners TA group, n= 34, 40kmweek, for over two years, aged 23 to 50 years, were instructed not to exercise for
24 hours before blood collection. Healthy sedentary subjects were included as controls CT group, n= 30. Adjustment was made
for age. Populations of CD34+, CD133+ cells and CD34+CD133+ subpopulations CPCs, CD34+KDR+ EPCs were
quantified by flow cytometry in the peripheral blood and represented as percentage of positive events in selected mononuclear
cells in the lymphocyte population % CMNL. The endothelial function was assessed by FMD of brachial artery using high
resolution vascular ultrasound. Data not normally distributed were compared by the Mann Whitney U test and Spearmans
correlation coefficient r and expressed as median standard error of mean. Data following normal distribution were analyzed
by the Student ttest and the Pearsons correlation coefficient, expressed as mean standard error of mean. P values < 0.05 were
considered significant. Adjustment for age was performed by a multivariate analysis. No statistical difference between groups
was found in the CPC counts: CT: CD34+: 0.0381 0.0036 %CMNL; CD133+: 0.0308 0.0043 %CMNL; CD34+CD133+:
0.0149 0.0024 %CMNL. TA: CD34+: 0.0399 0.0050 %CMNL; CD133+: 0.0194 0.0058 %CMNL; CD34+CD133+:
0.0118 0.0037 %CMNL. Higher levels of EPCs p<0.05 were observed in the TA group CD34+KDR+: 0.0135 0.0027
%CMNL compared to CT group 0.0083 0.0012 %CMNL. The FMD in the TA group 7.79 0.59% was similar to CT
7.90 0.59% with no correlation with EPC percentage.
Conclusions:
The longterm endurance exercise training leads to a recruitment of EPCs to the circulation. This finding was not correlated with
any changes in endothelial function evaluated by FMD, which could be explained by the fact that the studied population is
healthy. More research are needed to elucidate what are the underlying mechanisms involved in the mobilization of EPCs in
response of training in healthy individuals and its function in this context.
Keywords: endurance training, endothelial progenitor cells, endothelial function
Financial Support: CNPq, CAPES and FAPES.
Resumo:03-073
EARLY MODERATE RUNNING EXERCISE BLOCKED OBESITY INDUCED BY LITTER SIZE REDUCTION.
Ribeiro, T. A. D. S. 1; Tfolo, L. P. 2; Mendes, F. C. V. 2; Fabricio, G. S. 2; Malta, A. 2; Oliveira, J. C. D. 1;

Barella, L. F. 2; Martin, J. M. 1; Miranda, R. A. 2; Rinaldi, W. 1
1
Departament of Cellular Biology and Genetic , UEM
2
Departament of Physical Education , UEM
Objectives:
Nutritional insults during perinatal life can cause long-term changes in metabolism, which allow the development of obesity in
adult life. These alterations are associated to a disruption in the central nervous system that leads to an imbalance in the
autonomic nervous system. As the uterine phase, lactation is a sensible period to brain growth and exercise is a potent stimulus to
improve the metabolism. This work studies whether early moderate and low frequency exercise could be a deprogramming factor
in rats that become obese due to overnutrition during lactation.
After birth, the litter were divided into two groups: control group of nine pups (NL) and small litter group patterned with three
animals, this Animals were weaned at 21-day-old and were submitted to treadmill training during 70 days, 3 times/week (NLEXE and SL-EXE). The other two groups were formed by sedentary animals (NL-SED and SL-SED). Training was realized in
dark, always at 19h30min, with crescent time and speed (10-60 min and 10-20 m/min, respectively). At 90-day-old, rats were
submitted to a glucose tolerance test and the electrical activity of superior parasympathetic and sympathetic ganglion nerves were
recorded. Results: SL-rats were heavier (13%) and bigger (2%) than NL ones. The SL-rats had a fat tissue accretion of 2-fold.
SL-rats ate 12% more chow than NL-rats. Vagus electrical activity of SL-rats were 35% higher when compared to NL-rats, while,
sympathetic nerve of SL-rats showed 21% decrease in the electrical activity compared to NL-rats. Early exercise autonomic
nervous system in SL-rats.
Conclusions:
Early moderate running training deprograms the obesity induced by litter size reduction. Exercise decreased the fat pads,
improved the glucose intolerance and kept the balance of autonomic nervous system. Exercise reverted the imbalance of training
was able to improve the metabolism, increasing the glucose tolerance and block the obesity onset in SL-rats.
Keywords: MODERATE RUNNING EXERCISE , LITTER REDUCTION, ACTIVITY NERVOUS PARASYMPATHETIC,
ACTIVITY NERVOUS SYMPATHETIC
Financial Support: CNPq, CAPES and Fundao Araucria
Resumo:03-074
SKELETAL MUSCLE BLOOD FLOW AND OXIGENATION DURING MAXIMAL INCREMENTAL DYNAMIC
EXERCISE IN INDIVIDUALS WITH METABOLIC SYNDROME
Sales, A. R. K. ; Silva, B. M. ; Neves, F. J. D. ; Rocha, N. G. ; Medeiros, R. F. ; Pereira, F. D. S. ; Garcia,

V. P. ; Bertoldi, J. ; Manzzano, F. ; Nbrega, A. C. L. D.
Fisiologia e Farmacologia/Federal Fluminense (Biomdico), UFF
Objectives:
Dynamic exercise has been used to assess integrative cardiovascular, respiratory and muscular function both in healthy
individuals and in patients with exercise intolerance. Individuals with metabolic syndrome (MS), even in the absence of overt
cardiopulmonary disease, have lower peak oxygen uptake (VO2peak) during maximal incremental dynamic exercise when
compared with healthy individuals. In addition, these individuals have endothelial and microvascular dysfunction. Therefore, it is
conceivable that one of the mechanisms leading to their low VO2peak is impairment in muscle blood flow and oxygenation
during exercise. Given this background, the aim of the present study was to compare skeletal muscle blood flow and oxygenation
during maximal incremental dynamic exercise among individuals with SM.
Seven individuals with SM (23.0 2.0 years, BMI: 34.12.0) and 4 healthy controls (CT) (29.0 3.0 years, BMI: 22.10.8) were
enrolled. The individuals with MS had at least three of the following five criteria: waist circumference 90 cm (man) or 80 cm
(woman); triglycerides 150mg/dL; HDL cholesterol < 40 mg/dL (man) or < 50 cm (woman); systolic blood pressure 130
mmHg or diastolic blood pressure 85mmHg; fasting glucose 100mg/dL. The individuals of the CT group did not have any of
these criteria. The individuals performed a maximal incremental dynamic exercise test on an electronically braked cycle
ergometer. The protocol consisted on a linear increase of work (watts) every min with constant cycling rate (60 rpm). Ventilation
and expired gases were measured breath-by-breath and VO2peak was considered the highest oxygen uptake value obtained in the
last minute of exercise. Total hemoglobin (THb), an index of muscle blood, deoxyhemoglobin (HHb) and oxyhemoglobin
(HbO2) were measured in the vastus lateralis muscle (VLM) using near-infrared spectroscopy (NIRS). The NIRS probe was
positioned longitudinally on the belly of the VLM (~ 15cm above the patella) and secured with velcro straps around the thigh,
after the skin had been carefully shaved and cleaned. No movement of the probe was observed during exercise. This study was
approved by the Institutional Ethics Committee (013/2010). Individuals with SM had lower VO2peak than CT individuals (SM:
21.9 1.8 vs. CT: 36.03.0 mL.Kg-1.min-1, p<0.05).
Conclusions:
Individuals with SM had impaired skeletal muscle blood flow and oxygenation during exercise, which may be partially
responsible for the reduced VO2peak during maximal incremental dynamic exercise in these individuals.
Keywords: dynamic exercise , endothelial dysfunction, metabolic syndrome , peak oxygen, skeletal muscle
Financial Support: CAPES, CNPq, FAPERJ, FINEP, LABS`DOR
Resumo:03-075
THE CONCENTRATION OF A CYSTEINE PROTEASE INHIBITOR INCREASES IN SALIVA FOLLOWING AN

ANAEROBIC POTENCY TEST
Sant'anna,m. L. 1,3; Casimiro-lopes,g2; Sorenson,m. M1; Salerno,vp2

1
Instituto de Bioqumica Mdica, UFRJ
2
Escola de Educao Fsica e Desportos, UFRJ
3
Centro de Instruo Alte Sylvio de Camargo, CIASC
Objectives:
Physical exercise generates multiple physical and biochemical changes in the human body that can be monitored in various
manners. The measurement of these changes can be good indicators of workload, redox state and the state of the immune system.
However, the efficient evaluation of these changes is dependent on the variation measured and the approach applied. A need
exists for a rapid non-invasive approach to evaluate the impact of exercise on an individual. To evaluate the protein composition
of the saliva and identify proteins that display changes between rest and an anaerobic exercise (RAST running anaerobic sprint
test) identification by mass spectrometry.
The procedures in this study were approved by the Ethics Committee of the Hospital Universitario Clementino Fraga Filho. The
subjects were military pentathlon team athletes from the Brazilian Navy. The team consisted of five men and two women (age
27.1 2.3 years, weight 65.2 2.6 kg, BMI 21.9 0.3 kg/m2 and 10.1 2.2% fat). The RAST test was applied with the athletes
performing six 35-m maximal sprints with a 10-second interval between each sprint. Saliva was collected from each team
member before and 5 min after the anaerobic test, then the saliva was centrifuged and the supernatant was stored at 20 C. The
total protein concentration was determined by a Bradford assay. The concentration of protein in the saliva increased 2 fold
following the RAST (pre-test 0.62 0.03 mg / mL; post-test 1.30 0.08 mg / mL). The samples were normalized for total protein
concentration during sample preparation for SDS-PAGE and separated on an 4-18% gradient gel. Proteins in the gel were
visualized by Coomassie G and scanned for densitometry. Multiple bands displayed consistent differences between the before
and after test samples. A single protein band at 16 kDa displayed a large increase (202 16%, n = 7) in intensity from samples
collected after RAST. The band was excised from a preparative gel, digested with trypsin and analyzed by mass spectrometry
(ESI-Q-TOF). Three isoforms of a cysteine protease inhibitor were identified: cystatin SN (16.5 kDa), cystatin AS (14 kDa) and
cystatin S (14.4 kDa).
Conclusions:
Monitoring of the saliva is a promising rapid non-invasive approach for evaluating the impact of exercise on individuals. The
saliva displayed multiple proteomic variations including three cysteine protease inhibitor isoforms whose concentration increased
following anaerobic exercise.
Keywords: exercise, protease, inhibitor, cystatin, RAST
Financial Support: FAPERJ, CNPq, CIASC, Unit for Mass Spectrometry and Proteomics (CCS-UFRJ)
Resumo:03-076
MAXIMAL POWER OUTPUT ESTIMATES THE MLSS INTENSITY IN CYCLE ERGOMETER BEFORE AND
AFTER AEROBIC TRAINING
Wilke, C. F. ; Ramos, G. P. ; Fonseca, T. R. ; Mortimer, L. . C. F. ; Lima, A. M. ; Barros, C. L. M. D. ;
Mendes, T. T. ; Silami-gracia, E.
Federal University of Minas Gerais, UFMG
Objectives:
The maximal blood lactate steady-state intensity (MLSS) is considered the gold standard method for determining lactate
threshold, aerobic capacity evaluation and training prescription. However, the method is expensive and demands practice and
knowledge to data collection and blood handling. Aim: To establish an equation to predict MLSS intensity through a validated
VO2max incremental test protocol in cycle ergometer.
Method: The study was approved by UFMG ethic comitee (ETIC 153/08). Twenty-six physically active men (age 24 3y, body
mass 73.46.7kg, VO2max 46,9 6,6 mL.kg-1.min-1) were randomly divided into two groups (G1 and G2, n=13). All subjects
performed an incremental test until fatigue to determine the maximum power output (WMAX) and VO2MAX (25W.2min-1)
cycling at 50rpm and two to five constant intensity tests lasting 30 minutes (CT) to determine MLSS. In CT, MLSS was
considered the greater intensity of 30min cycling without blood lactate increase (0.05) (r=0.70; p0.05) (r=0.79; p<0.05).
Conclusions:
Conclusion: The proposed equation (WMFEL-EST = 0.866xWMAX+41.73) was able to estimate the MLSS power output before
and after six weeks of aerobic training. Therefore it would be possible to use only one test to obtain both measured VO2max and
estimated WMFEL in young, physically active individuals before and after an aerobic training period.
Keywords: estimation, lactate threshold, training
Financial Support: CNPq, FAPEMIG, CAPES , Ministrio do Esporte, FINEP, FUNDEP/SANTANDER.
Resumo:03-077
ACUTE AND CHRONIC EFFECTS OF EXERCISE ON THE VASCULAR REACTIVITY TO ISCHEMIA IN
HEALTHY WOMEN
Barbosa, T. C. ; Pereira, F. S. ; Rocha, N. G. ; Medeiros, R. F. ; Sales, A. R. K. ; Silva, B. M. ; Neves, F. J. ;

Nobrega, A. C. L.
Depto. Fisio. Farmacologia/Universidade Federal Fluminense, UFF
Objectives:
The endothelium mediates the interplay between the blood and the vascular wall, synthesizing and releasing many vasoactive
substances, which are crucial for vascular biology, including smooth muscle tone regulation. One of the ways that has been
employed to evaluate the endothelial responsiveness is through the temporary arrest of the forearm circulation, followed by a
prompt restoration of blood flow. Such ischemic stimulus leads to a great and sudden increase in shear stress, which increases
nitric oxide production by up to 200% above resting values and provokes a large reactive hyperemia. Previous studies have
shown that the vascular reactivity to ischemia is acutely enhanced after a bout of exercise in healthy subjects. However, it is still
unknown whether the acute enhancement of vascular reactivity after a bout of exercise would be changed by exercise training.
Therefore, the aim of the present study was to investigate the acute and chronic effects of exercise on the vascular reactivity to
ischemia in healthy subjects.
Ten healthy, sedentary, non-obese and non-smokers women (age 289 years, BMI 23.23.6 kgm2) were evaluated to investigate
the acute effect of exercise on the vascular reactivity. Their vascular reactivity was assessed at baseline, 10 and 60 min after an
acute bout of exercise (maximal exercise test) or a control resting period. Other 54 healthy, sedentary, non-obese and nonsmokers women (age 318 years, BMI 25.03.1 kgm2) were evaluated to investigate the chronic effect of exercise on the
vascular reactivity at baseline, 10 and 60 min after an acute bout of exercise. Thirty-nine of these women were submitted to
exercise training while 15 remained sedentary. The vascular reactivity was considered the change in forearm vascular
conductance [forearm blood flow (measured by venous occlusion plethysmography) divided by mean blood pressure (measured
by auscultation)] provoked by 5 min of circulatory occlusion. The maximal exercise test was performed on a treadmill until
voluntary exhaustion. The exercise training consisted of aerobic and resistance exercises performed 3 times per week, during 12
weeks. Vascular reactivity increased and remained elevated until 10 min after the exercise test [baseline: 7.01.2 vs. 10 min:
9.90.9 mL(100mL of tissueminmmHg), p0.05 vs. baseline]. There was no change in vascular reactivity during the control
session (p>0.05), in which no exercise was performed. Exercise training increased VO2peak [before: 30.00.8 vs. after: 35.01.0
mL(kgmin), p0.05]. On the other hand, at 10 min after the exercise test the vascular reactivity was enhanced after exercise
training [before: 9.40.6 vs. after: 10.70.7 mL(100mL of tissueminmmHg), p0.05).
Conclusions:
A single bout of exercise acutely increased the vascular reactivity to ischemia in healthy women, and this phenomenon was
enhanced after a period of exercise training.
Keywords: Acute effects, Aerobic training, Chronic effects, Maximal exercise test, Vascular reactivity
Financial Support: FAPERJ, CNPq, CAPES, FINEP
Resumo:03-078
ANTIOXIDANT RESPONSE IN SALIVA IN FIFTEEN MINUTES BICYCLE TEST
Viana Gomes, D; Rosa, F. L. ; Cardoso dos Santos, G. R. ; Navegantes, C. ; Ramos, K. ; Oliveira, L. T. ;

Real-hohn, A. H. ; Sanntanna, M. D. L. ; Salerno, V. P.
LABMOV/Universidade Federal do Rio de Janeiro, UFRJ
Objectives:
Exercise is known to increase energetic demands and the increased metabolic response can cause oxidative stress. Oxidative
stress can be characterized by an imbalance between the production of free radical and the buffering by antioxidants. Free radical
production has been observed to increase after acute exercise (Int J Dermatol. 40:747, 2001). The monitoring of free radical
production during exercise could be used to prevent a loss in athletic performance by avoiding systemic damage caused by free
radicals. To assess the bodys response to free radicals produced during a bout of intense exercise, the levels of antioxidants were
measured in saliva, which is easy to collect and requires no invasive procedures. The aims of this study were to assess the use of
saliva for measurements and to determine the antioxidant response during an acute bout of high intensity exercise in healthy
subjects.
The cohort consisted of six, healthy men aged 22 3.3 years. The high intensity exercise was consisted of cycling for 15 min
with a workload between 80 and 90% of maximal heart rate (HRmax). Saliva was collected before and after exercise. The levels
of lactate were measured in the rest and immediately after the activity using a lactimeter (Acctrend plus). The reduced glutathione
(GHS) and 1,1-Diphenyl-2-Picrylhydrazyl (DPPH) in saliva were measured following the protocol described by Look (Eur J Clin
Nutr. 51:266, 1997) and Janaszewska (Scand J Clin Lab Invest.62:231, 2002), respectively. The heart rate was measured using a
frequency meter (Garmin Forerunner 305). All experimental procedures were approved by the ethics committee of the
Clementino Fraga Filho Hospital UFRJ. The mean HR was 173 5.5 bpm, which corresponded to 82% of the maximum HR
for this age group, indicating that the exercise was high-intensity exercise and was confirmed by the concentration of lactate in
the blood. The lactate concentration at rest was 1.6 0.1 mM and immediately after exercise increased approximately 5 fold (9.2
3.3 mM), indicating a predominance anaerobic exercise. After the exercise, DPPH was decreased by approximately 13% (P >
0.05). The DPPH reduction could be observed because of an increase of antioxidants in the saliva. GHS, one of the main
antioxidants, increased around 10% in comparison with the rest (P = 0.005).
Conclusions:
The results indicate that this high intensity acute exercise was capable of inducing a change in the redox state in measured in the
saliva. While exercise increases the free radical production, healthy subjects are capable of buffering these reactive compounds
by increases in antioxidants system. Furthermore, the use of saliva is a viable system for measuring antioxidant capacity.
Keywords: Oxidative-stress, exercise, ANTIOXIDANT capacity, Free radical, Lactate
Financial Support: FAPERJ, CNPq
Resumo:03-079
EFFECT OF HIGH NUTRIENT AVAILABILITY ON REDOX BALANCE AND INSULIN RESISTANCE IN MUSCLE
CELLS
Sampaio, I. H. 1,2; Barbosa, M. R. 2; Silveira, L. R. 1,2

Escola de Educao Fsica e Esporte de Ribeiro Preto, EEFERP - USP
2
Bioqumica e Imunologia da Faculdade de Medicinada Rib Preto, FMRP - USP
1
Objectives:
Aim: We investigated the effect of elevated nutrient availability in the redox state of cells from skeletal muscle and relation with
insulin resistance.
Methods and Results: The skeletal muscle cells were isolated from quadriceps muscle from Wistar rats and cultured in DMEM
medium. After differentiation, the cells were exposed to different concentration of nutrients including glucose (5 mM), glucose
(10 mM), glucose (25 mM) and fatty acid (palmitic acid 700 M) with or without insulin (10.000 U/mL). The muscle cells were
also previously treated with N-Acetylcysteine (NAC) (10 mM) or electrically submitted to moderate muscle contraction (60 min).
The oxygen consumption was measured using a Clark-type electrode (Hansatech Instruments). The intracellular reactive oxygen
species (ROS) production, NAD+/NADH ratio and GSH/GSSG ratio were determined by fluorimetric assay. In addition, the
mRNA level of PGC1 was determined by PCR-Realtime and uptake of 2-desoxy-[2,6 3H]-glucose (0.1 Ci/mL) by
radioactivity assay (Scintillator Beckman-LS 5000TD). The oxygen consumption was significantly reduced at presence of
palmitate plus insulin (p<0.05).
Conclusions:
Conclusion: Our results showed that elevated nutrient availability reduces mitochondrial oxygen consumption favoring ROS
production and consequently insulin resistance. The effect of antioxidant and muscle cell contraction shifting the intracellular
redox state suggest that ROS are important regulator of glucose metabolism in skeletal muscle cells.
Keywords: High Nutrient, Redox Balance, Insulin Resistance, Muscle Cells
Financial Support: Support: FAPESP, PIBIC (USP) and CNPq.
Resumo:03-080
EVALUATION OF THE EFFECT OF MODERATE PHYSICAL TRAINING ON THE BEHAVIOR OF THE
VELOCITY OF CONDUCTION IN DIABETIC RATS
Silva, R. T. B. 1; Lira, K. D. S. 1,2; Figueiredo, T. G. 1; Maia, J. N. 1,3; Moraes, S. R. A. 1,2,3

1
Department of Anatomy , UFPE
2
Program of Post-Gradation in Physiotherapy , UFPE
3
Program of Post-Graduation in Neurosciences and Behavioral S, UFPE
Objectives:
This study investigated if a regular moderate aerobic physical training could influence the development of diabetic neuropathy
peripheral.
We used 18 male rats, albino, Wistar, with body weight ranging between 230-270g (animals were induced to diabetes). They
were separated into: Sedentary control group (SCG, n=4); Sedentary diabetic group (SDG, n=4); Trained control group (TCG,
n=5) and Trained diabetic group (TDG, n=5). After fasting of 12 hours at 60 days of life the animals of the diabetic groups were
induced to diabetes (streptozotocin solution). The trained groups were submitted to a protocol of moderate physical training,
swimming 5 days/week for 6 weeks, and duration from 10 minutes (initially) to 60 minutes (the end of the protocol). The sensory
nerve conduction velocity (SNCV) was checked at the beginning and end of the study. Weekly we checked body weight and
serum glucose of the animals. The independent samples were submitted to analysis of variance - ANOVA (one way) followed by
Bonferroni test, and the paired samples were submitted to Student t test. The results were expressed as mean standard deviation
with a confidence level of 95%. At the end of the experiment, there wasnt difference in SNCV between the experimental groups
(SCG = 43,222,50; SDG = 38,610,87; TCG = 40,143,95; TDG = 39,204,25), but there was a decrease in body weight of
diabetic groups (SDG = 216,00a23,21 and TDG = 193,20b41,25) in comparison to other two groups (SCG = 355,00a55,73 e
TCG = 256,00b57,57), and serum glucose of TDG (303,40c86,70) in comparison with the SDG (369,50c44,49), but this
reduction was not sufficient to normalize glucose levels.
Conclusions:
The results of this study suggest that exercise training may protect the peripheral nervous system by lowering blood glucose
levels, thereby reducing nervous exposition to high glucose levels.
Keywords: NERVE CONDUCTION VELOCITY, DIABETES MELLITUS, SWIMMING
Financial Support: CNPq/PIBIC/UFPE e CNPq/Universal n 477096/2008-5.
Resumo:03-081
HYPOTENSION AFTER RESISTANCE EXERCISE OF DIFFERENT INTENSITIES IN HYPERTENSIVE WOMEN:
RESPONSE OF THE FOREARM BLOOD FLOW
Brito, A. F. 1,2,3,4; Santos, M. S. B. 1,2,3,4; Santos, A. D. C. 1,2,3,4

Programa Associado de mestrado em Educao Fsica UPE/UFPB, UPE/UFPB
2
Lab of Research in Physical Training, Performance and Health, LETFADS
3
University of Pernambuco/Escola Superior de Educao Fsica, UPE/ ESEF
1
Dept of Physical Education/ University Federal of Paraba, DEF/ UFPB
Objectives:
The physiological responses promoted by resistance exercise minutes after their execution are controversial and have not been
sufficiently clarified, especially in hypertensive public, considering that it is treated by different drug classes. For this, we
evaluated the effect of one session of resistance training with different intensities in the post-exercise hypotension (PEH) and
forearm blood flow (FBF).
The study was conducted with ten women with hypertension (55 3 years; 25.7 3 kg/m2), who regularly participated in a
program of resistance exercise at the UFPBs gym. To participate in the study, they should be carrying only hypertension as
cardiometabolic disease and use only the drug class of inhibitors of angiotensin converting enzyme and diuretics. To determine
the sample size used the 3.1.0 software Gpower a statistical power of 0.80, an alpha error of 0.05, a difference of PEH for systolic
blood pressure of 2 mmHg and residual standard deviation of 2 mmHg. The women were subjected to three experimental
sessions: session control (SC), exercise 50% (EX50%), and 80% (EX80%) of 1 RM, ever held between seven and nine o'clock in
the morning. The order was determined randomly and individually so that each subject has its own order. Before the study, all
were instructed to avoid foods high in caffeine 12 hours before the collections and do not perform physical activities 48 hours
before the experimental sessions. For each session, subjects were assessed pre-and post-intervention. In the pre-intervention,
women were at rest in the supine position for 10 minutes and at the end of this was recorded BP (by Finometer) and FSA
(through the plethysmograph, Hokanson AG 201, Washington, USA). Later they were taken to the gym, where they remained for
about 20 minutes to the sessions of exercise (EX50% and EX80%) or remained at rest in each of the equipment during the same
time in the session SC. EX50% and EX80% were composed for a set of 10 repetitions in 10 exercises, with an interval of 90
seconds between exercises. Then, the women returned to the laboratory, were placed in the supine position for the FSA and BP
measurements were again performed at 10, 30, 50, 70 and 90 minutes of recovery (post-intervention). The PEH was more evident
in EX80% compared to EX50%, where its magnitude was -28mmHg versus -18mmHg for SBP, -13mmHg versus -8mmHg for
DBP and -27mmHg versus -18mmHg for MAP, respectively. As a result, the FSA has increased significantly in both sessions,
but this increase was most evident in the EX80% at 10, 30, 50, 70 and 90 minutes of recovery (4.97 0.28, 4.73 0.28, 4.88
0.29; 4.81 0.50; ml/min/100ml 4.80 0.25) compared with EX50% in the same moments, respectively (4.22 0.24, 4.10
0.38; 4:36 0.27, 4.28 0.28, 4.10 0.39 ml/min/100ml).
Conclusions:
Thus, we conclude that resistance exercise was effective in causing PEH, this phenomenon is accompanied by increases in FSA
within the first minute of recovery.
Keywords: hypotension, resistance exercise, blood pressure, hypertension, forearm blood flow
Resumo:03-082
CHRONIC ABSENCE OF TAIL ARTERY INNERVATION ATTENUATES CUTANEOUS HEAT LOSS AND
INCREASES BLOOD PRESSURE DURING PHYSICAL EXERCISE UNTIL FATIGUE.
Lima, M. R. M. 1; Pires, W. 1; Fonseca, I. A. T. 1; Wanner, S. P. 1,2; Coimbra, C. C. 1,2; Lima, N. R. V. 1

1
Department of Physical Education, UFMG
2
Department of Physiology and Biophysics, UFMG
Objectives:
Tail skin vasodilation is the principal mechanism of heat loss during physical exercise in rats, and cutaneous blood flow is
modulated by vasoconstrictor sympathetic activity. Denervation of tail artery acutely increases heat loss, a response that is
blocked after a few days as a consequence of morphological and functional adaptations of the tail vasculature. So far, there are no
published reports of the effects evoked by chronic absence of tail artery innervation on exercise-induced thermoregulatory and
cardiovascular responses. Therefore, the present study was aimed at investigating the effects of chronic absence of the tail artery
innervation on cutaneous heat loss and cardiovascular adjustments during physical exercise until fatigue.
All experimental procedures were approved by the Ethics Committee for Animal Experimentation of the Federal University of
Minas Gerais (protocol 109/09). Adult male Wistar rats were used in the experiments (210-240 g; at the time of first surgery).
Under anesthesia, the animals were submitted to tail artery denervation (TAD) or Sham-TAD surgery as control procedure. After
3 weeks of recovering, temperature sensors were implanted into the peritoneal cavity and polyethylene catheters were inserted
into the ascending aorta. Each rat had two days to recover and was then submitted to two experimental situations: (1) exercise on
a treadmill at 18m/min until fatigued (Sham-TAD: n=6 and TAD: n=10) or (2) resting on the treadmill belt (Sham-TAD and
TAD: n=5). Both experimental situations were performed at the same time of day with an interval of two days. During the
experiments, core body temperature (Tb), tail skin temperature (Ttail), mean arterial pressure (MAP), and heart rate (HR) were
measured. Glyoxylic acid-induced fluorescence was used to confirm the effectiveness of denervation. Data are expressed as
means S.E.M. Differences between groups were assessed by a multiple factor analysis of variance followed by the least
significant difference test. Paired Students t-test was used to compare physical performance. Significance level: P < 0.05. In
resting rats, TAD did not change the thermoregulatory and cardiovascular parameters. In contrast, TAD attenuated the exerciseinduced increase in Ttail (Sham-TAD: 33.3 0.6 C vs. TAD: 31.5 0.4 C; fatigue point; p=0.03). Despite this change in
cutaneous heat loss, neither Tb nor time to fatigue was different between groups. Furthermore, TAD rats presented a greater
exercise-induced increase in MAP (Sham-TAD: 124 3 mmHg vs. TAD: 133 2 mmHg; fatigue point; p=0.03), although HR
was not different compared to Sham-TAD animals.
Conclusions:
Our results indicate that the chronic absence of tail artery innervation is determinant for cutaneous heat loss and cardiovascular
adjustments during physical exercise until fatigue. Furthermore, we suggest that the attenuation of heat loss in denervated-rats
may have resulted from morphological or/and functional adaptations in tail vasculature, and that the absence of reflex bradycardia
response was possibly due to changes in the baroreflex sensitivity.
Keywords: cardiovascular system, performance, thermoregulation
Financial Support: CNPq, CAPES, and FAPEMIG
Resumo:03-083
CHARACTERIZATION OF S-NITROSOGLUTATHIONE REDUCTASE IN SKELETAL MUSCLE AND
PHARMACOLOGICAL INHIBITORS
Yamashita, A. M. S. ; Mousinho, V. C. ; Figueiredo-freitas, C. ; Sorenson, M. M.

Objectives:
Objectives: The purpose of this research is to characterize the expression and activity of GSNO-R in skeletal muscle and to
examine its role in the regulation of muscle contraction.
Methods: The enzymatic activity of GSNOR was measured at 340 nm by consumption of GSNO in homogenates of rat liver,
kidney, brain, heart, soleus (type I fibers) and extensor digitorum longus (EDL, type IIb fibers). Activity was normalized to total
protein concentration (Pierce BCA protein assay kit). Expression of GSNOR was characterized using electrophoresis followed by
Western blot with a polyclonal antibody. Results: The activity of GSNO-R was detected in all tissues tested. Activity found in
skeletal muscle was similar to that in other tissues such as heart and brain. The GSNO-R inhibitor C1 (3 - [1 - (4-acetylphenyl)-5phenyl-1H-pyrrole-2yl] propanoic acid) was tested at 30 M, pre-incubated with the homogenate for 2 min. It inhibited GSNO-R
activity in all tissues.
Conclusions:
Conclusions: The enzyme GSNO-R has enzymatic activity in skeletal muscle similar to tissues such as heart and brain, with
minor variations depending on the major types of skeletal muscle fibers in soleus and EDL.
Keywords: glutathione, muscle, Nitric oxide, S-nitrosoglutathione reductase, S-nitrosylation
Financial Support: CNPq, FAPERJ
Resumo:03-084
MITOCHONDRIAL COMPLEXES ALTERATION IN ANIMALS EXERCISED AT HIGH INTENSITY INTERVAL
TRAINING
Ricardo, J. C. ; Martins, E. L; Ramos D. ; Casimiro-lopes, G; Jardim-messeder, D; Salerno, V. P; Sorenson,

M. M; Galina, A.
Universidade Federal do Rio de Janeiro, UFRJ
Objectives:
Previous studies suggested that the benefits of high intensity interval training (HIIT) compared with endurance exercises are
related to increased in expression of oxidative enzymes and mitochondrial biogenesis. In addition, rats subjective to HIIT showed
decreased insulin resistance and increased GLUT4 in cell membrane. However, analyses of mitochondrial physiology in
glycolytic and oxidative muscles were not described. The objective was evaluate the mitochondrial physiology in oxidative and
glicolitic muscles from animals subjective to HIIT.
One group of 5 male Wistar rats (Rattus norvegicus) was sedentary (S) and the other trained by swimming to non-exhaustion for
3 days for 2 weeks (T). The animals were sacrificed 48 hours after of the last training session. Soleus and tibialis anterior muscles
were isolated and the fibers were permeabilized with saponin to evaluate mitochondrial oxidative activity with different
substrates. There were no changes in physical capacity, in total body mass and in visceral fat mass. However, contrary to
expectations, there were observed a 1.5 fold decrease in the maximal oxygen consumption for both tibialis anterior (glycolytic)
and soleus (oxidative) muscles.
Conclusions:
Contrary to expectations proposed for HIIT we observed that the mitochondrial maximal respiration capacity in muscle is
decreased. This result suggests a muscle adaptation to anaerobic exercise.
Keywords: mitochondrial complexes , exercised , high intensity training
Financial Support: CNPq and FAPERJ
Resumo:03-085
EFFECTS OF CHRONIC ABSENCE OF ARTERIAL BARORECEPTORS ON BLOOD PRESSURE AND HEART
RATE VARIABILITY DURING PHYSICAL EXERCISE PERFORMED IN THE HEAT.
Pires, W. ; Lima, M. R. M. ; Wanner, S. P. ; Fonseca, I. A. ; Vaz, G. F. ; Fumega, U. ; Coimbra, C. C. ;

Lima, N. R. V.
Educao Fsica / Universidade Federal de Minas Gerais, UFMG
Objectives:
There is evidence showing that chronic absence of arterial baroreceptors disrupts the sympathetic-cardiovascular responses to
heat stress. The spectral power analysis of blood pressure (BPV) and heart rate (HRV) variability has been used to evaluate the
modulation of the autonomic nervous system during several physiological conditions, including the physical exercise. Thus, the
present study was aimed at investigating the effects of chronic absence of arterial baroreceptors on blood pressure and heart rate
variability during exercise performed in a warm environment.
Adult male Wistar rats weighing 250-350 g were used in all experiments. Animals were housed in individual cages under
controlled light (0500-1900 hours) and temperature conditions, with water and rat chow provided ad libitum. All experimental
procedures were approved by the Ethics Committee of the Federal University of Minas Gerais for the Care and Use of Laboratory
Animals (protocol 178/10). The animals were submitted to sinoaortic denervation (SAD; n=5) or sham denervation (SHAM; n=5)
as a control procedure. After two weeks, a polyethylene catheter was implanted into the ascending aorta, and a temperature sensor
was implanted into the peritoneal cavity. Rats were allowed to recover during two days and were then submitted to the exercise
trials on a treadmill at 18 m/min until fatigued. Each animal ran in both cool (25 C) and hot (35 C) conditions on separated
days. Pulsatile arterial pressure was recorded (sample rate = 2 KHz) by the aortic catheter and intraperitoneal temperature (Tb) by
telemetry. Power spectral density was obtained using the fast Fourier transformation method and Hanning windows (512) with
50% overlap. Spectral power components for very low- (VLF, < 0.05. During exercise in the heat, SHAM rats showed higher
BPV (LF; 11.4 2.6 mmHg2 SHAM-Hot vs. 5.0 0.3 mmHg2 SHAM-Cool; p<0.05).
Conclusions:
The effects evoked by the chronic absence of arterial baroreflex suggest that the mechanism plays an important role modulating
autonomic nervous system and core temperature adjustments induced by physical exercise performed in the heat.
Keywords: baroreceptors, blood pressure, exercise, spectral analysis, temperature
Financial Support: CAPES, CNPq and FAPEMIG.
Resumo:03-086
A NEW METHOD TO STUDY FATIGUE DURING EXERCISE: RAT WHEEL RUNNING DURING A
PROGRESSIVE DISTANCE/FOOD SYNCHRONIZED PROTOCOL.
Fonseca, I. A. T. 1; Arajo, F. D. A. 2; Lima, M. R. M. 1; Pires, W. 1; Young, R. J. 3; Rodrigues, L. O. C. 1

1
Department of Physical Education , EEFFTO/UFMG
2
Physiologic Science, ARFIS/UFU
3
Zoology, PUC/MG
Objectives:
In wild environment, animals exercise for food, reproduction and to escape from predators. In laboratory rats, exercise has only
been obtained by the unnatural electrical stimulation. The rat daily food achievement synchronized to an exercise bout has not
been investigated yet. Therefore, the aim of this study was to measure the rat wheel activity pattern during a progressive
distance/food ratio (DFR) daily protocol, to simulate a natural variability in food resources in the wild environment.
Five male Wistar rats at 6 weeks of age (228 1.4 g, mean SEM) were housed individually in a metal cage (GAUSTEC
LTDA) (40 x 40 x 40 cm) with a metal wheel (diameter 30 cm, circumference 942 mm). They had free access to wheel, food and
water for 10 days (baseline wheel running situation). In baseline situation the spontaneous running activity was 1422 484.9
m/day and the ad libitum food ingestion was 23 0.7 g/day. Then the running wheel was connected to an electronic food
dispenser (distance/food situation) that released a food pellet (about 4.1 0.1g) synchronized to the running distance. Within this
device, the animal must exercise to receive food. Initially, the first synchronized distance/food ratio (DFR in m/g) was the
average distance run and food intake in baseline situation (66 m/g). From this situation the DFR was arbitrarily increased
progressively (15% every three days for 72 days). The total distance run, average speed, delivered pellets were recorded each
hour during the whole experiment. Body mass and food intake were measured every day between 7 am to 11 am. The food intake
was calculated by the difference between delivered and eaten pellets. The animals were kept in a controlled light (14 - 10 h lightdark cycle) and temperature (24 - 26 C) environment. Data are expressed as means SEM. Running distance, food intake and
body mass were compared between situations using ANOVA followed by the Tukeys test. The correlation between rat running
distance and DFR was done using Pearsons correlation coefficient. The significance level was set at p < 0.05. The rats mean
running distance during exercise synchronized with food dispenser situations increased progressively according to the increase in
DFR (114 35.4 m/g in baseline situation to 309 11.7 m/g in the last situation; p < 0.05). The animals increased their running
distance to obtain the daily food amount increasing their time spent in exercise for food. The average speed was not different
between the DFR situations but increased when the food was synchronized in the first situation in comparison to baseline (17
2.1 vs. 27 0.4 m/min; p< 0.05) and then there were no further differences.
Conclusions:
The new method created a rat voluntary exercise that seems to be closer to its natural activities in wild conditions.
Keywords: distance, fatigue, food, voluntary exercise, wheel running
Financial Support: FAPEMIG; CNPq; CAPES.
Resumo:03-087
THE EFFECT OF SIBERIAN GINSENG STANDARDIZED EXTRACT ON BIOCHEMICAL PARAMETERS,
WEIGHT AND PERFORMANCE IN SEDENTARY RATS TESTED FOR TREADMILL RUNNING.
Arouca, A. B. 1; Crege, D. R. X. D. O. 1,1; Ramos, L. A. F. 1; Ishizu, L. Y. 1; Faria, L. O. M. D. 2; Areas, M.

A. 2; Costa, K. G. D. 8; Silva, F. O. C. D. 8; Macedo, D. V. D. 9; Grassi-kassisse, D. M. 1
1
Depto. de Anatomia, Biologia Celular, Fisiologia e Biofsica, UNICAMP
2
Depto. de Anatomia, Biologia Celular, Fisiologia e Biofsica, UNICAMP
3
Depto de Anatomia, Biologia Celular, Fisiologia e Biofsica, UNICAMP
4
5
6
7
Depto. de Bioqumica, UNICAMP
8
9
10
Objectives:
This study aimed to examine whether Siberian ginseng standardized extract (Eleutherococcus senticosus) promotes effect on
weight, biochemical parameters such as glucose, lactate, triacylglycerol, cholesterol and performance in sedentary rats submitted
to an incremental treadmill Performance Test (PT).
Methods and materials: Adult male Wistar rats (n = 12) were divided into two groups (control, C and supplemented, S). Animals
were adapted to treadmill running test (10 m/min) for 15 days through 10 minutes/day. After this period, animals were submitted
to the first PT (Test 1): 3 minutes in a speed of 10m/min; after that, increase 1m/min every 2 minutes until speed 20m/min; after
that, increase 2m/min every 3 minutes until exhaustion. The S group received oral Siberian ginseng standardized extract diluted
in 1mL of 0.9% saline solution (100mg/kg, 5 days per week) while the rats in group C received only 1mL 0.9% saline solution
for 8 weeks. Weight and food intake were controlled. The groups were submitted to other two PT: after the fourth week (Test 2)
and after the eighth week (Test 3). Through a small incision at the tail, we analyzed the parameters of lactate in Accutrend
Performance test strips (Roche) and blood glucose Active test strips (Roche) before, immediately after and 5 minutes after the
end of PT, considering Test 1 and Test 3. The animals were sacrificed 48h after Test 3. Biochemical parameters of blood glucose,
triglyceride and cholesterol were analyzed before sacrifice by Accutrend Plus strip (Roche) and Active strip (Roche).
Performance analysis was evaluated by MATLAB program, in which rat mass and running time inside PT were considered and
the results were expressed as rat mass x meter (kg x m). One-way ANOVA and Tukey-Kramer test for multiple comparisons
were used for statistical analysis. Student t test was used for additional analysis (blood glucose, cholesterol and triacylglycerol).
Significantly results were considered when p 0.05). There were no significant differences in biochemical parameters like blood
glucose (C, 129.83.40 mg/dL), cholesterol (C, 158.02.30 mg/dL) and triacylglycerol (C, 210.521.66 mg/dL) (p>0.05).
Conclusions:
The present data indicated that the oral administration of Siberian ginseng standardized extract in sedentary rats was effective in
improving performance in a treadmill PT along 8 weeks even though no metabolic changes occurred.
Keywords: Performance, Siberian ginseng, Treadmill running, Wistar rats
Resumo:03-088
DIFFERENT CHONIC EXERCISE PROGRAMS AFFECT THE BONE MINERAL CONTENT AND DENSITY IN
OVARIECTOMIZED FEMALE RATS
Fernandes, B. B. 1; Drummond, L. R. 1; Peluzio, M. C. G. 1; Del Carlo, R. J. 1; Silva, C. H. O. 1; Castro, C.

A. 1; Jnior, J. F. Q. 1; Ramos, R. M. S. 1; Lavorato, V. N. 1; Natali, A. J. 1
1
Depto. de Educao Fsica/Universidade Federal de Viosa, UFV
2
Depto Nutrio e Sade Humana/Universidade Federal de Viosa, UFV
3
Depto de Medicina Veterinria/Universidade Federal de Viosa, UFV
4
Depto. de Estatstica/Universidade Federal de Viosa, UFV
Objectives:
The deficiency secretion of estrogen in women in the post-menopausal state is related to bone mass loss. This study investigated
the influence of chronic treadmill running and swimming on the bone mineral content (BMC) and density (BMD) in
ovariectomized female rats.
Female Wistar rats (age: 20 wks; body weight: 271,42 17,6g) underwent surgical ovariectomy (OVX) or laparotomy (SHAM)
and were allocated to OVX running (OR), SHAM running (SR), OVX swimming (OS), swimming SHAM (SS), OVX control
(OCON), or SHAM control (SCON). Animals were housed in individual cages in a room with temperature of 22 2C and
light/dark cycle of 12 hours. They received 20g of chow daily had water ad libitum. Animals in the running groups underwent a
program of treadmill running (16m/min, 60min/day, 0 grade, 5 days/week) for 10 weeks. Animals in the swimming groups
swam in a tank filled with water (30 2 C) with an overload of up to 3% of body weight (60min/day, 5 days/week) for 10
weeks. Animals in OCON and SCON groups remained their cages without scheduled exercise for 10 weeks. After euthanasia the
right femur was removed, cut free of soft tissues for further. The BMD and BMC analysis were performed using DEXA
equipment. ANOVA three way was used to compare BMC and BMD among groups and a significance level of p< 0,05) body
mass than that of SHAM animals by the end of the experimental period. The SR group had higher BMD compared to SS (0.237
0.015 g/cm2 vs. 0.219 0.018 g/cm2, respectively). The animals in the OS group exhibited a higher BMD than those in the SS
group (0.241 0.029 g/cm2 vs. 0.219 0.018 g/cm2, respectively). The OR group showed higher values of BMC than OCON
group (0,308 0,026 g vs. 0,272 0,036 g, respectively).The OS group presented higher BMC as compared to OCON (0,301
0,054 g vs. 0,272 0,036 g, respectively). The values for BMC in the SCON group were higher than those in the OCON group
(0.297 0.032 g vs. 0.272 0.036 g, respectively).
Conclusions:
Chronic swimming increased the BMD and BMC in OVX female rats whereas chronic treadmill running increased the BMC in
OVX and the BMD in SHAM female rats.
Keywords: Physical activity, Menopause, Bone Health, Rats, Osteoporosis
Financial Support: FAPEMIG
Resumo:03-089
RESISTANCE TRAINING REGULATES IL-10 AND SREBP-1C EXPRESSION, SIZE AND WEIGHT OF
MESENTERIC FAT MASS IN OVARIECTOMIZED RATS
Stotzer, U. S. ; Duarte, F. O. ; Gatto, C. D. V. G. ; Silva, G. H. G. D. ; Domingues, M. M. ; Rodrigues, M.

F. C. ; Perez, S. E. A. ; Arajo, H. S. S. D.
Cincias Fisiolgicas/ Universidade Federal So Carlos, UFSCar
Objectives:
The exercise has prophylactic effect upon the pathogenesis triggered by chronic inflammation. This benefic effect seems to be, in
part, result of the reduction of visceral fat promoted by regular exercise and/or an induction of anti-inflammatory cytokines after
each exercise session. Visceral fat mass is an important source of interleukin 10 (IL-10), an anti-inflammatory cytokine which is
increased in ovariectomized rats. However, the anti-inflammatory role of resistance training is poorly explored and is unknown in
ovariectomized rats. Therefore, we investigated the effect of both one bout and chronic resistance training on the IL-10 mRNA
levels in ovariectomized rats. In addition, we investigated the expression of SREBP-1C mRNA levels, a transcription factor
involved in lipogenesis and the size and weight of mesenteric fat mass.
Adult female Sprague-Dawley rats were randomly divided into six groups (n=6 per group): sham-sedentary (S-SED),
ovariectomized-sed (OVX-SED), S-trained (S-T), OVX-T, S-acute (S-A) and OVX-A groups. Sham or ovariectomy procedure
was performed at 10 weeks of age. Three weeks after the surgery, trained groups started 10 weeks of climbing in a 1.1-m vertical
ladder with progressive load of 65%, 85%, 95% and 100% of rats previous maximal overload, plus 30g attached to their tails
until exhaustion. S and OVX acute groups performed one bout session. All animals were killed 92 days after ovariectomized.
Trained animals were killed 48 hours after the last training session. Immediately, mesenteric adipose tissue was weight and part
of this tissue was immediately frozen in liquid nitrogen for mRNA analyses and 100mg was prepared for morphological analyses
in colidina and osmium tetroxide. The IL-10 mRNA expression was significantly lower (64%) and SREBP-1C was higher (36%)
in OVX compared to sham groups (p
Conclusions:
These data highlight that resistance training can help in reducing the usual state of chronic inflammation in ovariectomy-induced
obesity by decreasing SREBP-1C expression and visceral fat area as well by increasing the gene expression of the antiinflammatory cytokine, IL-10. These results may be clinically relevant to the menopausal population.
Keywords: Estrogen, Exercise, Gene expression, Adipose cells
Financial Support: CNPq and CAPES
Resumo:03-090
INFLUENCE OF EXERCISE ORDER ON MUSCLE DAMAGE IN TRAINED MEN
Gattai, P. P. ; Ferreira, J. C. ; Wurtele, M. ; Arajo, R. C. ; Foschini, D.

Biofsica, UNIFESP
Objectives:
Analyze the effects of change in the usual order of strength training exercises on muscle damage, number of repetitions and
delayed onset muscle soreness (DOMS) in young trained adults.
We selected 10 healthy young adults (age= 22.142.67 years) with at least 6 months experience in strength training leading to
muscle hypertrophy (BMI= 25.622.88 kgm). Subjects were divided randomly into two groups: A control group (CG), which
performed the exercises in the usual order of training (ChestTriceps) and an experimental group (EG), which performed the
exercises in the reverse order. The study design was initiated by a session of strength tests (triceps machine and bench press
machine) to determine the load of maximum voluntary contraction (MVC) in both exercises. The following week, the volunteers
performed eight sets of each exercise at 75% MVC. Subjects were allowed to pause for 1 minute between the bouts and for 3
minutes between exercises. We considered increases in plasma creatine kinase (CK) and a reduction in the number of repetitions
(RNR) between subsequent sets as an indicative of muscle damage. Muscle soreness was assessed using a subjective perception
of pain scale. 3 blood samples, the first 30 minutes before the training session, another immediately after the session and a third
24 hours after the same session were taken. For statistical analysis we used the Student t test and ANOVA. The CK levels
increased 24 hours after the training session only in the CG. The total volume of training conducted by the CG was higher than
that undertaken by EG. The RNR between sets was higher in the EG when compared to the CG for both exercises.
Conclusions:
After data analysis, we reach the conclusion that the execution order of strength training exercises influences the volume of work
performed as well as the muscle damage, but not the DOMS, and that furthermore the group that initiated with the smaller muscle
group exercise had a lower workload during the training session.
Keywords: Delayed onset muscle soreness, Exercise Order, Muscle damage, Strength training
Financial Support: Conselho Nacional de Desenvolvimento Cientfico e Tecnolgico
Resumo:03-091
RECOVERY PROCESS OF THE ILEUM CONTRACTILE RESPONSE AND REDOX STATUS AFTER ONE
SESSION OF MODERATE EXERCISE IN C57BL/6 MICE
Mendona, K. M. ; Berro, L. F. ; Martinez Jr, G. ; Alves, G. A. ; Silva, L. R. ; Ribeiro, R. F. ; Aboulafia, J. ;

Nouailhetas, V. L. A.
Biofsica, UNIFESP
Objectives:
Exercise is an important tool in prevention and treatment of chronic-degenerative diseases. Despite several evidence of the
beneficial effects of exercise, it has been shown that an acute session of moderate aerobic exercise causes increased production of
free radicals, which may induce oxidative stress, with consequent damage to many organs and tissues. The aim of this work was
to study the effects of an acute moderate aerobic exercise on the contractility and level of oxidative stress in the intestinal muscle
of C57BL/6 mice mainly by focusing on the recovery process.
Three-month old, male C57BL/6 mice were submitted to an acute session of treadmill running at 55-65% Vmax after 5-days of
acclimatation period. Animals were ascribed to the following groups: control (CT), exercised and killed immediately after the end
of the exercise session (EX), exercised and killed after 12, 24, 48, 72, 96 and 120 hours after the end of the exercise session
(REC-12, REC-24, REC-48, REC-72, REC-96 and REC-120, respectively). Data are presented as percentages in relation to
control group. We performed Students t test and one-way ANOVA (followed by Bonferroni post-test) to compare groups. P <
0.05 was considered statistically significant. Ileum isometric contractions were recorded in the presence of Tyrode solution at
37C, pH adjusted to 7.4, bubbled with air. Tissue responsiveness was evaluated determining the potency (EC50) and the efficacy
(Emax) from non-cumulative concentration-contractile response curves built up in response to either carbachol (CCh), bradykinin
(BK) or KCl-induced membrane depolarization. EC50 was determined as the concentration of the stimulant that induced 50% of
the maximum contraction (Emax). Emax values from KCl-induced contractions were significantly decreased in REC-12 (40%),
REC-24 (33%), REC-48 (33%), REC-72 (39%), REC-96 (42%), REC-120 (36%) relative to CT group. Also, Emax values from
CCh-induced contractions were significantly decreased in REC-12 (27%), REC-24 (29%), REC-48 (30%), REC-72 (48%), REC96 (52%), REC-120 (53%) relative to CT group. Finally, Emax values from BK-induced contractions were significantly
decreased only in REC-24 (54%), REC-96 (53%), REC-120 (48%) relative to CT group. Oxidative damage to whole ileum
proteins was spectrophotometrically quantified by determining tissue carbonyl content according to the method described by
Reznick and Packer (1994).and spectrophotometrically assessed at 370 nm. Protein content of the last pellet was evaluated
through Bradford (1976) technique. We observed a significantly greater carbonil quantity in REC-72 (56%) in comparison with
CT, and EX, REC-96 and REC-120 groups did not differ from the control group.
Conclusions:
It is shown that one acute session of moderate exercise impaired both the eletro- and pharmacomechanical couplings in C57Bl/6
intestine. Interestingly, even with a recovery time raging from 12 to 120 hours, this altered contractility is not recovered. The
increased level of protein oxidation was evident only 72 hours after the end of exercise session suggesting that ileum may suffer
oxidative stress, despite the moderate intensity of the exercise protocol. Shorter recovery periods should be analysed in order to
better characterize the recovery process.
Keywords: isometric contraction, moderate exercise, oxidative stress, protein oxidation
Financial Support: Fapesp
Resumo:03-092
PRIOR AEROBIC EXERCISE TRAINING PARTIALLY PREVENTS SCIATIC NERVE INJURY-INDUCED
SKELETAL MUSCLE DYSFUNCTION IN RATS
Gomes, K. M. S. ; Bechara, L. R. G. ; Vessoni, A. T. ; Campos, J. C. ; Moreira, J. B. N. ; Voltarelli, V. A. ;

Mattos, K. C. ; Monteiro, A. W. A. ; Brum. , P. C. ; Ferreira, J. C. B.
Escola de Educacao Fisica e do Esporte/ USP, EEFE-USP
Objectives:
Introduction and objective: Sciatic nerve injuries are known to quickly induce severe skeletal muscle atrophy and dysfunction,
while aerobic exercise training (AET) has been shown to improve skeletal muscle function in healthy and sick individuals.
Considering that potential protective effect of AET against sciatic nerve injuries have never been described before, we aim in the
present study to characterize the effects of previous AET on skeletal muscle mass and ex vivo function in rats underwent to
sciatic nerve injury (SNI).
Methods: Twenty-nine male Wistar rats were randomly assigned into the following groups: sedentary control submitted to fake
surgery (Sham; n=5), sedentary submitted to SNI (S-SNI; n=10) and aerobically trained submitted to SNI (T-SNI; n=14). It is
worth highlighting that AET was performed prior to SNI surgery. All rats performed a graded treadmill exercise test until
exhaustion before and after AET protocol. Moderate-intensity AET was performed during four weeks (60% of maximal running
speed; five days/week). Rats were killed 14 days after Sham or SNC surgery and EDL muscle was carefully harvested and
weighted. Ex vivo EDL muscle function was evaluated in an organ tissue bath equipped with a force transducer, where muscles
were mounted and electrically stimulated while incubated with Krebs Heinselet solution. Supra-maximal stimulation current and
optimal muscle length were determined (1 Hz) with a constant resting period (60s) between subsequent contractions. Following a
10-min recovery period, forcefrequency characteristics (10, 20, 30, 50, 80, 100 and 150Hz; 350 ms) were recorded. Peak
isometric force during electrical stimulation was registered. Data are presented as mean standard error from mean. Statistical
significance was considered achieved when P was
Conclusions:
Conclusion: Taken together, our data suggest that previous moderate-intensity AET reduces EDL muscle atrophy and contractile
dysfunction in rats submitted to SNI.
Keywords: AEROBIC EXERCISE TRAINING, SCIATIC NERVE INJURY, SKELETAL MUSCLE
Resumo:03-093
FAT PARTITIONING EFFECTS OF HIGH INTENSITY INTERVAL TRAINING IN DEXAMETHASONE TREATED
RATS.
Martins, E. L. 1; Marinho, G. M. 1; Souza, F. 1; Chicaybam, G. 2; Barreto, A. 2; Pires, L. 2; Ramos, D. M. 1,2;

Casimiro-lopes, G. 1,2
1
2
Instituto de Nutrio, UERJ
Objectives:
High intensity interval training (HIIT) is a training method whose main characteristic is based on short periods of exercise bouts
with small rest intervals between sets completed in few minutes of duration when compared with traditional training protocols.
HIIT method is able to promote many health benefits, however this protocol is exhaustive and not adequate as a nonpharmacological therapy. However non-exhaustive bouts of HIIT also showed some results control rats suggesting that this could
be a promising strategy for health promotion. Dexamethasone (DEXA) treatment can induce insulin resistance, altered lipid
profile and other associated diseases in rats. The reproducibility of this model could be an adequate opportunity to evaluate the
effects of HIIT method in some alterations induced by DEXA. The objective of this study was to evaluate the effects of nonexhaustive bouts of HIIT in rats treated with dexamethasone.
Adult male Wistar rats (Rattus norvegicus) were divided in four groups: Sedentary (S; n=5), Exercised (EX; n=5),
Dexamethasone (D; n=5) and Dexamethasone exercised (D-Ex; n=5). The treatment with DEXA was done during eight days.
Following that period EX and D-Ex rats were subjected to three non-exhaustive bouts of HIIT protocol. The exercise sessions
were done in a swimming pool and consisted of 3 bouts of 20 seconds with 10 seconds of rest while bearing a weight equivalent
to 10% of their body weight. The training during 8 days alternated. The treatment with DEXA was maintained during the exercise
training and 48 hours after the last training session all the animals were sacrificed with a lethal dose of sodium thiopental (5mg/
100g). Statistical analysis was done with ANOVA one-way with Newman-Keuls post test, with p value set at 0.05. Both DEXA
treated rats (D and D-Ex) presented significantly lower total body mass (-34% and -26% respectively). Visceral fat mass (-61% in
both groups; p
Conclusions:
Non-exhausive bouts of HIIT protocol were efficient to reverse TAG accumulation in liver of DEXA treated rats. Longer periods
of training could offer additional benefits in another health parameters. Our results suggest that this king of training could be a
promising strategy as a non-pharmacological therapy in fat altering conditions as a less time consuming alternative.
Keywords: Exercise, Dexamethasone, Triacylglycerol in tissue
Resumo:03-094
ACUTE AEROBIC SWIMMING EXERCISE INCREASE NUCLEOTIDE CATABOLISM IN RAT BLOOD SERUM
Senger, M. R. 1,2; Pedrazza, E. L. 3; Oliveira, . R. D. 2; Bonan, C. D. 3

2
ESEF, UFRGS
1
Instituto Oswaldo Cruz, FIOCRUZ
3
Programa de Ps-Graduao em Biologia Celular e Molecular, PUCRS
Objectives:
Besides its consolidated role as an energetic molecule, ATP has many functions in extracellular space. At the cardiovascular
system this molecule and its catabolics, mainly ADP and adenosine, are involved in the blood coagulation and in the vascular
tone, functions that are strongly altered during physical activity. The sequential hydrolysis of ATP to adenosine its realized by a
group of enzymes called nucleotidases. This group of enzymes is constituted by NTPDases, 5-nucleotidase and NPP. The
purpose of this investigation was to examine the effect of an acute swimming exercise session on nucleotide catabolism on rat
blood serum.

Male wistar rats were divides in two groups (n=6 each group): exercise (E) and sedentary (S). The group (E) the rats was
submitted to one swimming session (60 min) with a constant load of 4% of body weight in the tail. The rats of the (E) group
presented a significant increase (student t-test, P0.05) on the activity of the three enzymes tested when compared to (S) group.
The ATP and ADP hydrolysis were increased 55.3% and 43.1%, respectively. The 5-nucleotidase was activated 57.4%on the (E)
group. The NPP activity was activated 24.2%.
Conclusions:
Our results has shown that the ectonucleotidases are activated after one session of aerobic swimming exercise in rat blood serum,
suggesting that this enzymes are involved in an increased adenosine production and consequently an exercise-mediated
vasodilatation.
Keywords: Exercise, Nucleotidases, Serum, ATP, Adenosine
Financial Support: CAPES, FAPERJ, CNPq.
Resumo:03-095
ROLE OF NAD(P)H OXIDASE ON TENSION PRODUCTION CAPACITY AND FATIGABILITY OF ISOLATED
RAT SKELETAL MUSCLE
Vessoni, A. T. ; Bechara, L. R. G. ; Guimares, F. L. ; Negro, C. E. ; Brum, P. C. ; Ramires, P. R.

Escola de Educao Fsica e Esporte da Univ. de So Paulo, EEFEUSP
Objectives:
To investigate, in an in vitro model, the involvement of the enzymatic complex NAD(P)H oxidase on contractile function of
skeletal muscle tissue at rest and during contractile activity.
After being sacrificed by decapitation, the right and left soleus muscles of 7 male Wistar rats (14 month old, 485 39g) were
carefully removed and attached to an isometric force transducer within an in vitro system containing aerated (95% O2, 5% CO2)
Krebs-Ringer solution (in mM: 137NaCl, 5KCl, 2CaCl2, 1KH2PO4, 1MgSO4, 24NaHCO3, 11C6H12O6; 22oC; pH 7.4).
Following a 30-minute stabilization period, in which individual optimal lengths for isometric contraction were determined (L0,
184.4 24.3 mm), the muscles were electrically stimulated (0.2 ms and 80 v pulses) 3 times, with increasing train durations and
pulse frequencies (1 ms at 1 Hz; 300 ms at 30 Hz; 1200 ms at 100 Hz). Then, they randomly underwent one of two possible
pharmacological treatments: incubation with 1mM apocynin a NAD(P)H oxidase inhibitor (APO, n=7) or distilled H2O (CON,
n=7) during 20 minutes, before being re-stimulated (as described above) to determine the effects of inhibition of enzymatic
complex NAD(P)H oxidase on the capability of skeletal muscle to generate isometric tension under basal conditions. The results
presented as the mean standard deviation of range (%) of tensions generated by each group before and after incubation were
compared using the Students t-test with p < 0,05. After 30 minutes of incubation, the muscles were then submitted to a low
frequency fatigue-inducing protocol, in which they were stimulated during 8 minutes with 300 ms trains at 30 hz, separated by
900ms intervals, in order to determine the effects of inhibition of enzymatic complex NAD(P)H oxidase on the tissue fatigability.
The results presented as mean standard deviation of variation (%) of the initial tension generated by each group after 60
second-intervals were compared using a 2-factor (time, treatment) variance analysis (ANOVA) with p < 0.05. Under basal
conditions, apocynin-treated muscles showed a greater ability to generate isometric tension under submaximal electical
stimulations of 1ms at 1Hz (0.18 0.08% vs. 0.09 0.07%, p=0.05) and 300ms at 30Hz (0.17 0.10% vs. 0.05 0.08%, p =
0.03), when compared to H2O-treated group. However, when submitted to a maximal electrical stimulation (1200 ms at 100 Hz),
the isometric tension generated in both groups was not influenced by apocynin (0.01 0.02% vs. 0.02 0.04%, p=0.30).
Regarding fatigue, the muscles treated with apocynin showed greater losses of capacity to generate isometric tension after 6 (0.48
0.12% vs. 0.59 0.08%, p < 0.05), 7 (0.42 0.12% vs. 0.54 0.10%, p < 0.05) and 8 (0.36 0.11% vs. 0.50 0.10%, p
Conclusions:
The inhibition of NAD(P)H oxidase by apocynin promoted an increase in the ability of isolated skeletal muscle to produce
submaximal isometric tension under rest, although the same treatment accelerated the fatigue process of the tissue when subjected
to an increased contractile activity.
Keywords: apocynin, fatigue, isometric force, skeletal muscle, oxidative stress
Financial Support: CNPq and FAPESP.
Resumo:03-096
EFFECTS OF WESTERN DIET AND MODERATE-INTENSITY EXERCISE ON THE COLONIC OXIDATIVE
STRESS.
Haddad, M. T. 1; Guimares, C. B. D. 1; Papineli, R. 1; Ribeiro, R. F. 2; Nouailhetas, V. L. A. 2; Rosa, E. F.

1,2
1
Centro Universitrio So Camilo, CUSC

Departamento de Biofsica/Universidade Federal de So Paulo, UNIFESP
Objectives:
A Western diet (WD), defined by high-fat, low-calcium and vitamin D content has been shown to induce colonic oxidative stress.
Recently we observed that moderate exercise practiced throughout life could prevent the increase of intestine oxidative stress.
Since these factors have been related to the development of colon cancer and the effects of moderate exercise and western diet on
the colonic redox status are poorly known, the aim of this study was to evaluate their impact on colonic oxidative stress.
Male C57BL/6J mice, 4 wk old, were ascribed into four groups, two of them fed with AIN-76A standard diet: sedentary (CTSED, n=4) and exercised (CT-EX, n=5); and two others fed with western diet: sedentary (WD-SED, n=5) and exercised (WDEX, n=10). Animals in WD-SED and WD-EX groups had access to the AIN-76A standard diet from weaning until the 4th week
of life, followed by free access to the western diet ad libitum, until animal death, while animals from CT-SED and CT-EX groups
had free access ad libitum to the standard AIN-76A diet. The amount of feed ingested by mice from WD-SED and WD-EX was
measured daily throughout the procedure, and body weight control was done at 7 days interval throughout the procedure. Body
weight from CT-EX group was measured before the onset of exercise. Animals from exercised groups were submitted to 5 days
of adaption which consisted of a daily session of treadmill running at 10m/min for 15 min. Animals from CT-EX and DI-EX
groups performed a moderate exercise protocol consisiting of treadmill running at 13m/min, which started at 12th week of the
animal life and terminated 48 hour before the animal death Sacrifice of the animals in all groups has occurred by cervical
dislocation at 16 weeks of life. Heart and gastrocnemius muscle from each animal were isolated, weighed, and expressed as g/g
body weight for determining the level of cardiac and gastrocnemius hypertrophy. The level of colon oxidative stress was
determinate by the level of protein oxidation, measuring carbonyl radicals by the technique of Reznick. Animals submitted to a
Western Diet presented a mortality rate around 37,5 %, during the treatment, while control animals mortality rate was around 1%.
Animal body weight from CT-EX group increased of 6.6% at the onset of moderate exercise, while the body weight in WD-SED
and WD-EX groups increased of 29,2% and 22,2%, respectively after the initiation of the diet. Food intake varied over the weeks.
Animals in WD-SED group start eating 10.3 0.5 g/animal in the first week, and ended up ingesting 15.7 0.0 g in the last
week; animals from WD-EX group ingested 11.0 0.2 g/animal in the first week and 16.2 g in the last one. No influence of either
the moderate exercise or the western diet was detected in the level of cardiac and gastrocnemius hypertrophy and in the level of
carbonyl radicals formed.
Conclusions:
The Western diet increased the mortality rate of animals. However, Western Diet and physical exercise have no impact on
intestinal protein oxidation of colon.
Keywords: Intestine, Moderate-exercise, Oxidative-stress, Western Diet
Financial Support: FAPESP, Centro Universitrio So Camilo
Resumo:04-025
EFFECT OF PH ON GLUCOSE AND ELECTROLYTE JEJUNAL ABSORPTION IN RATS
Viana, M. P. ; Borges, E. L.
Dpto de Biofsica e Fisiologia/Instituto Cincias Biolgicas, ICB-UFMG
Objectives:
Aim: Studies have shown that glucose is able to decrease the pH of the surface epithelium jejunal preparations when added in
vitro and the existence of a high concentration of protons in the immediate area of the mucosa could be of considerable
significance for absorption of electrolytes (J. Nutr. 116: 768, 1986). The aim of this study is to assess whether the change in pH
of Tyrode (solution used for perfusion of the jejunum) interferes with the absorption of glucose and electrolytes.
Methods and Results: Male Wistar rats weighing 200 to 220g (n=6) were utilized. All experiments were carried out in compliance
with the local Ethical Principles in Animal Experimentation (CETEA/UFMG protocol no 230/2010). Jejunal absorption of
glucose and electrolytes was investigated in rats. A Tyrode solution containing twice glucose, sodium and potassium
concentration (pH 7.0, 7.4, 8.0 and 8.5) was infused through the jejunal loops during 40 minutes. The glucose absorption was not
significantly affected by Tyrode pH. However, there was significantly decrease in sodium absorption at pH 7.0 and 8.5 (41.13
2.79 mM and 41.37 1.71 mM, respectively; p<0.05).
Conclusions:
Conclusion: These data indicate that the pH of Tyrode has no influence on glucose absorption. However, the major potassium
uptake occurs at pH 8.0, while the absorption of sodium is impaired at pH 7.0 and 8.5.
Keywords: Absorption, Glucose, pH, Potassium, Sodium
Resumo:04-026
THE INFLUENCE OF CORTICOSTERONE IN THE DIFFERENTIATION OF MUCOUS NECK CELLS IN RAT
GASTRIC MUCOSA DURING EARLY WEANING
Zulian, J. G. ; Osaki, L. H. ; Rigonatti, C. A. M. ; Gama, P.

Instituto de Cincias Biomdicas, USP
Objectives:
Recent studies suggest that mucous neck cells (MNC) are markers of cellular differentiation in the gastric epithelium. Early
weaning (EW), that means the abrupt interruption of suckling, increases differentiation in the rat gastric mucosa, in parallel with
high corticosterone levels. This hormone can direct or indirectly regulate gastric growth and maturation. Thus, our aim was to
investigate whether corticosterone is involved in MNC differentiation.
Wistar rats were separated into four groups on the 15th postnatal day: suckling control (S), suckling treated with RU-486 (SRU),
early weaning (EW) and early weaning treated with RU-486 (EWRU). (Protocol approved by CEEA ICB n 86/08). In the EW
groups, the pups were separated from their dams and milk was substituted by semi-solid and solid diet. After the onset of early
weaning, pups were i.p. injected with vehicle (corn oil) or RU-486, an antagonist of glucocorticoid receptor. MNC population
was evaluated on day 18, in samples submitted to histochemical reactions with lectin Griffonia simplicifolia II (lectin GSII) and
also Periodic Acid-Schiff combined with Alcian Blue (PAS-AB). Early weaning increased the number of MNC evidenced by
PAS-AB compared to S group (P
Conclusions:
Our results showed that the blockage of corticosterone action by RU-486 administration impairs mucous neck cell differentiation
stimulus triggered by early weaning. We conclude that besides the dietary pattern, corticosterone also has a role in the control of
gastric cell differentiation.
Keywords: corticosterone, dietary pattern, differentiation, stomach, weaning
Financial Support: CAPES e FAPESP
Resumo:04-027
EFFECTS OF ANTI-CD3 ANTIBODY IN SALIVARY GLANDS OF SPONTANEOUSLY DIABETIC MICE
Metidieri, H. T. ; Mayoral, . E. ; Rojas, F. A. ; Peroni, L. A. ; Mncio, R. D. ; Picardi, P. K. ; Caldeira, E.

J.
Tissue Morphology Laboratory-Morphology and Basic Pathology, FMJ
Objectives:
Diabetes mellitus results in various complications, also compromising the salivary glands. The current treatment for this
condition should be a substituting method to exogenous insulin, which, in spite of considerable advances, is still associated to
constraints and lack of long-term effectiveness in relation the recovery of the tissue injuries. Thus, the aim of this study was to
evaluate the anti-CD3 antibody as alternative therapy in the recovery of salivary glands of spontaneously diabetic mice.
Ten spontaneously diabetic mice (NOD) were divided into two groups of 5 animals: group I (untreated diabetic mice) and group
II (anti-CD3/treated diabetic mice). The animals were kept under standard conditions of housing, feeding and treatment at the
Sector of Laboratory Animal Experimentation, Department of Morphology and Basic Pathology, Faculty of Medicine of Jundia,
FMJ. Group II received the anti-CD3 antibody (IMUNY BIOTECHNOLOGY) administered intravenously (5 g/day) by the
period of 5 days. Group I received daily intravenous injections of saline to simulate the experimental conditions of treated group.
Glucose level was monitored during the treatment and salivary gland samples were collected at the end of experimental period for
morphometrical analysis. Treated animals consumed less liquid (ml) (0.85 0.09 P 0.05) and solid (g) (-0.48 0.67 P 0.05),
when compared with the animals of the group I (1.9 0.45 and 0.63 0.13 respectively). The weight (g) was increased in
animals of group II (+ 0.32 0.22 P 0.05). High levels of glucose (mg/dl) were observed in untreated animals, while in treated
animals, the reduction was observed (165.6 29.87). Tissue restructuring (Statistical Results: 0.0209 P 0.05) was observed in
animals submitted to immune therapy.
Conclusions:
The immune treatment promoted the recovery of salivary acinar cells, demonstrating that this immunoregulatory function of antiCD3 might be important for the reversal of hyperglycemia-induced tissue injury.
Keywords: salivary glands, structure, diabetes, treatment
Financial Support: NAPED and FAPESP (grant number: 2010/05455-8 and 2010/51619-2)
Resumo:04-028
PROTECTION OF PORTAL HYPERTENSIVE GASTROPATHY BY ESTROGEN IN MODEL PARTIAL PORTAL
VEIN LIGADURE (PPVL)
Morgan-martins, M. I. 1; Hartmann, R. M. 2; Schemitt, E. G. 1; Marroni, N. A. P. 1

1
Universidade Luterana do Brasil, ULBRA
2
Hospital de Clnicas de Porto Alegre, HCPA
Objectives:
Portal hypertension (PH) is a complication secondary to cirrhosis that is characterized by an increased blood flow and / or
vascular resistance in the portal system, causing the appearance of a collateral circulation hyperdynamic. The partial portal vein
ligation (PPVL) is the experimental model used in rats to study the pathophysiological mechanisms involved in pre-hepatic portal
hypertension. Estrogen is an antioxidant molecule with various physiological actions. The aim of this study was to evaluate the
antioxidant activity of endogenous estrogen in experimental rats PPVL compared with intact rats castrated.
We used 20 Wistar rats, weighing on average 250g were divided into four groups: sham-operated (SO); intact with partial portal
vein ligation (IL), castrated (C) and castrated with partial portal vein ligation (CL). On Day 1: castration or sham-operated, on the
7th day of surgery PPVL, the 15th day after the LPVP was observed portal pressure (PP) in the mesenteric vein of rats on the
polygraph Letica. Blood was withdrawn by retro-orbital plexus to measure the level of estradiol an diferents groups. Was
evaluated oxidative stress, were the lipid peroxidation (LPO) in the stomach was assessed using the technique of thiobarbituric
acid reactive substances (TBARS) and activity of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT). For
histological evaluation the sections of the stomach were stained with haematoxylin and eosin. Statistical analysis was ANOVA Student-Newman-Keuls, (Mean SE) were considered significant at p
Conclusions:
In this experimental model, we observed that estrogen a contributed to preserve the gastric tissue and portal pressure maintained
at normal levels. Possibly by stimulating nitric oxide synthesis and decreased production of vasoconstrictor agents and reactive
oxygen species, decreasing lipid peroxidation.
Keywords: oxidative stress, estrogen, portal hypertension
Financial Support: FIPE
Resumo:04-029
ACTIVITY OF DIGESTIVE ENZYMES FROM THE HEPATOPANCREAS OF LITOPENAEUS VANNAMEI
SUBMITTED TO DIFFERENT DIETARY LEVELS IN HETEROTROPHIC CULTURE
Nascimento Jr, C. R. 1; Silva, J. F. 1; Andrade, D. H. H. 1; Souza, K. S. 1; Melo, F. P. 2; Verssimo, T. 2; de

Arajo, R. R. 2; Silva, J. F. X. 1; Correia, E. S. 2; Bezerra, R. S. 1
1
Universidade Federal de Pernambuco, UFPE
2
Universidade Federal Rural de Pernambuco, UFRPE
Objectives:
Carciniculture has been the main sector in the Brazilian production of aquatic organisms, especially in rearing Litopenaeus
vannamei. However, among the problems encountered by producers stand out spending with feed that is based on application of
fishmeal. In order to reducing costs and increase the shrimp production, various farming techniques are being researched such as
heterotrophic system that has shown promising results. Despite the success of this system, little is known of its effect on digestive
enzymes that have a significant influence on the determination of the nutritional requirements of animals.
the aim this work was to evaluate the activity of digestive enzymes of L. vannamei submitted to different dietary levels in
heterotrophic system. Specimens of L. vannamei weighing 1.100.23 g were reared in heterotrophic for a period of 12 weeks on
the Base de Pesca e Aquicultura (UFRPE). The food consisted of a commercial feed containing 35% crude protein and was
offered twice daily in the levels of 8% (control group T1), 6% (T2) and 4% (T3) of the total biomass of each tank. After the
cultivation period, were collected ten hepatopancreas of the shrimp of each treatment and homogenized in a solution of 0.01 M
Tris-HCl pH 8.0 and 0.15 M NaCl at a concentration of 40 mg tissue/mL and centrifuged at 10,000xg at 4 C for 25 min to obtain
the enzyme extract. The proteolytic activities were determined using the nonspecific substrate (1% azocasein) and specific
(BApNA, SApNA e Leu-p-Nan). The electrophoretic patterns (SDS-PAGE) and protease zymogram were carried out with
separation gels at a concentration of 12.5%. The use of 1% azocasein showed a higher total proteolytic activity in T2
(5.20.4U.mg-1). It was found higher trypsin activity to T2 (25.10.1U.mg-1) and T3 (24.51.7U.mg-1) using BApNA as
substrate. Analyzing the enzymatic action of chymotrypsin through the use of SApNA, it was found that the greatest activity was
in T3 (0.440.05U.mg-1). Regarding the activity of leucine aminopeptidase in the presence of the Leu-p-Nan, enzymatic activity
was higher for T1 (1.140.09U.mg-1) and T3 (1.080.04U.mg-1). The electrophoretic profile of the enzymes was 15, 17 and 18
protein bands, for the treatments T1, T2 and T3, respectively, with molecular masses of between 7 and 200 kDa. The proteolytic
activities of these bands were demonstrated in zymograms and animals of treatments T1 and T3 showed 10 bands with activity,
while T2 presented 13. In all treatments, it was noted bands that showed activity below 29 kDa.
Conclusions:
According to the results, it was observed that the heterotrophic culture positively affected the activity of digestive enzymes,
suggesting an improvement in digestion and assimilation of nutrients by L. vannamei. Also, show the ability of shrimp to adapt to
different growing conditions, especially in reducing the food supply, enabling it to feed on microorganisms in the water.
Keywords: heterotrophic, protease, Litopenaeus vannamei
Financial Support: CNPq, SEAP/PR, FINEP/RECARCINE, FACEPE e EMBRAPA
Resumo:04-030
EFFECTS OF FOOD RESTRICTION ON THE MYENTERIC PLEXUS AND MORPHOLOGY OF ILEUM OF
WISTAR RATS IN AGING PROCESS
Cirilo, C. P. ; Rampazzo, A. P. D. S. ; Schoffen, J. P. F. ; Zapater, M. C. V. U. ; Vicentini, F. A. ; Natali, M.

R. M.
Dep. Cincias Morfolgicas/ Universidade Estadual de Maring, UEM
Objectives:
The myenteric plexus is composed of neurons responsible for integrating the motor activity of the gastrointestinal tract and by
glial cells that play important roles in the maintenance of the neuronal homeostasis and regulation of intestinal epithelial barrier.
The objective of this work was to evaluate the total neuronal population, the glial cells and the elements of the ileum wall of rats
in aging process submitted to food restriction.
For this study, male Wistar rats (Rattus norvegicus) were divided into 3 groups (n=5), C7 and C12, euthanized at 7 and 12
months of age, respectively, and RA12 that received 50% of the feed given to the C12 group for 5 months (from 7 months of
age), being killed at 12 months of age. After anesthesia of the animals were collected the blood for biochemical analysis, and
samples of ileum, for histological study of the intestinal wall and obtaining whole-mount preparations subjected to HuC/D and
S100 immunohistochemical techniques for total population neuronal and glial cells analysis, respectively. The quantification was
performed in 64 images/animal/technique (intermediate and antimesenteric regions) and measurements made at 100 cellular
bodies/animal/ technique. For histological analysis, samples of ileum were fixed in Bouin and embedded in paraffin (sectioned to
7-m-thick) for the total wall, tunica mucosa and muscle analysis and in historesin (sectioned to 2.5-m-thick) for evaluate the
villous, crypts, goblet cells and metaphase index. The sections were stained by the histochemical reaction of periodic acid-Schiff
(PAS) to evidence the goblet cells and using hematoxylin-eosin for the other analysis. The determination of metaphase index and
quantification of goblet cells were accomplished in 2500 cells/animal. Morphoquantitative analyses of myenteric plexus and
morphometry of ileal wall were accomplished using the software Image Pro-Plus 4.5. The data were submitted to analysis of
variance (ANOVA) and Tukey's post test with a significance level of 5%. Comparing the C7 and C12 groups was possible to
detect that the aging process didnt altered blood components analyzed, the number and cellular profile of HuC/D and S100
immunostained cells. Histological analysis showed a reduction of muscular layer and metaphase index (p
Conclusions:
The food restriction was beneficial for reducing cholesterol and triglycerides levels. However, we considered that the model of
food restriction caused morphoquantitatives alterations in the myenteric plexus and in the histology of the ileum, interfering
negatively in intestinal function.
Keywords: aging, enteric glia, food restriction, intestinal wall, myenteric plexus
Financial Support: CNPq, Fundao Araucria
Resumo:04-031
THE EFFECT OF 1,8-CINEOLE ON GASTRIC EMPTYING RATE AND ON THE CONTRACTILE BEHAVIOR OF
GASTRIC OR DUODENAL ISOLATED STRIPS OF RATS.
Veras, H. R. F. ; Juc, D. M. ; Silva, M. T. B. ; Santos, A. A. ; Magalhes, P. J. C.

Departamento de Fisiologia e Farmacologia, UFC
Objectives:
1, 8-cineole is a monoterpene found in essential oils of plants found in the Northeast Brazil such as Croton nepetaefolius and
Eucalyptus tereticornis, which are commonly used in folk medicine for treating respiratory and gastrointestinal disorders. In this
study, we intended to evaluate the effects of 1,8-cineole on gastric emptying (GE) rate of a liquid test meal, as well as to
determine its effects on the contractile behavior of gastrointestinal smooth muscle in vitro.
Evaluation of the GE rate was performed using male Wistar rats (200-250g) that were divided in 3 groups: control (C; vehicle
0.2% Tween 80, p.o.) and 1,8-cineole-treated rats that received by gavage 10 (CIN10) or 100 mg/kg (CIN100). After 30 min, rats
received perorally a liquid test meal (Glucose 5%, Phenol Red 0.75 mg/ml). Ten minutes later, the animals were euthanized
following ethical guidelines, and visceral exeresis was performed to determine the fractional dye retention in rat stomach. The
amount of dye recovered in C group was 53 2% (n = 8), values significantly higher (p < 0.05, ANOVA) than those observed in
CIN10 (44 5%; n = 8) or CIN 100 (38 4%; n = 8) groups. In another set of animals, isolated strips of gastric fundus or
duodenum were maintained in bath chambers containing Tyrode solution (resting tension = 1 g; pH 7.4; 37 C, continuously
aerated). Recordings of the contractile activity were obtained through force transducers connected to data acquisition system
(Windaq, PM-1000, USA). Solutions of 1,8-cineole were prepared by adding Tween 80 (0.2% v/v). In a concentration-dependent
manner (p < 0.001, ANOVA), 1,8-cineole (0.1 to 3 mM) relaxed the basal tone of the strips of gastric fundus or duodenum with
EC50 of 1.09 [0.89 - 1.34] and 0.58 [0.25 -1.36] mM (n = 6), respectively. In preparations of gastrointestinal strips under Ca2+free conditions and in presence of EGTA (0.2 mM) and K+ (60 mM), 1,8-cineole inhibited the concentration-effect curve induced
by Ca2+ (0.1, 1 and 2mM) addition, reducing the contractile force induced by the addition of Ca2+ at 2 mM to 24.0 3.4% of a
reference contraction induced by 60 mM K+, values significantly lower (p < 0.05, Paired t-test) than those obtained in control
preparations (80.0 10.2%).
Conclusions:
1,8-cineole accelerates gastric emptying rate of liquids in rats and it possesses myorelaxant properties on isolated strips of gastric
fundus or duodenum. Such myorelaxant actions are putatively due to its ability in reducing Ca2+ influx through plasma
membrane in smooth muscle cells as previously reported (Clin Exp Pharmacol Physiol, 36(11):1120, 2009) and they are probably
involved in the 1,8-cineole ability to produce increased GE rate in rats.
Keywords: 1,8-cineole, gastric emptying, gastrointestinal smooth muscle, gastric fundus and duodenum, rats
Financial Support: CNPq
Resumo:04-032
THE SEROTONERGIC/OXYTOCINERGIC PATHWAY IS INVOLVED IN DECREASED GASTRIC EMPTYING
INDUCED BY ACUTE EXERCISE IN AWAKE RATS.
M. T. B da Silva1; W Okoba1; C. P. S. Campos 1; F. G. V. Oliveira1; A. D. N Pinheiro1; R. C. Palheta-junior

2
; A. A. dos Santos1
1
Department of Physiology and Pharmacology-UFC/Fortaleza-CE-, FISFAR
2
School of Veterinary Medicine-UNIVASF/Petrolina-PE,, CMVET-UNIVASF
Objectives:
In our previous publication, we showed that acute exercise exploiting the lactate threshold decreases gastric emptying of liquids
in rats. Thus, we decided to investigate the involvement of the serotonergic/oxytocinergic pathway on cardiovascular parameters
and gastric retention (GR) induced by acute exercise (AE) in rats.
After approval by the local ethics committee, N82/10, 48 male Wistar rats (230-280g)were equally divided into respective
groups Sham (S), Exercise (E), Sham+Ondansetron (SO), Exercise+Ondansetron (EO), Sham+Atosiban (Sa) and
Exercise+Atosiban (Ea). Initially, all rats were subjected to a non-weight aquatic adaptation (a scaled 10 to 40min exercise, for 5
days). After 24 hours, they were anesthetized for the cannulation of femoral blood vessels and thereafter fasted for over 24 hours
with free access to an oral rehydration solution (ORS). On the day of experiment, all rats underwent an exercise session, EA
(swimming for 15-min, 5% BW) with the groups SO, EO, Sa and Ea pretreated (15min.) with (Ondansetron 50g/kg or Atosiban
10g/kg via iv). Rats in group S were placed in shallow waters over 5 min. Consequently, we monitored the MAP and HR
parameters over the next 10-min. Shortly afterwards, the animals were fed to a liquid test meal (SG 5% phenol red) to assess the
GR, 10-min postprandial (J Physiol, 131:452-62, 1956). Data was expressed as meanSEM and compared by ANOVA and
Tukey test. Results: When compared to E, the S, Sa and Ea groups manifested hypotension (110.4 1.7, 106.26 3.6 and
87.11.2 vs 121.71.9 mmHg, p <0.001).
Conclusions:
AE provoked adjustments in cardiovascular parameters, thus increasing GR of liquids. This was prevented by prior treatment of
the animals with Ondansetron and Atosiban, confirming the involvement of the serotonergic/oxytocinergic pathway in this
phenomenon.
Keywords: gastric retention, acute exercise, serotonin, oxytocin
Financial Support: CNPq, CAPES and FUNCAP
Resumo:04-033
HEMODYNAMIC, ENDOCRINE AND GASTRINTESTINAL ACTIVITY INDUCED FROM RIGHT ATRIAL
STRETCH IN AWAKE RATS.
Junior, R. C. P. 1; Pinheiro, A. D. N. 2; Okoba, W. 2; Oliveira, F. G. V. 2; Silva, M. T. B. 2; Elias, L. L. K. 3;

Antunes-rodrigues, J. 3; Santos, A. A. 2
1
Universidade do Vale do So Francisco, UNIVASF
2
Fisiologia e Farmacologia/Universidade Federal do Ceara, UFC
3
Faculdade de medicina de Ribeiro Preto, USP-RP
Objectives:
Isotonic hypovolemia increases the gastric retention (GR) of a liquid meal, the central venous pressure (CVP) and plasmatic
oxytocin (OT) levels (Neurogastroenteroly 11:93, 1999; Exp Neurol 206:192, 2007). Thus, we decided to determine GR, plasma
levels of OT, as well as hemodynamic responses due to mechanical atrial stretch (AS).
After approval by the mandated Research Ethics Committee 2009/02, male Albino rats, (250-280g, n=56), were anesthetized and
submitted to right auricle appendectomy or not (control) before performing the cannulation of the carotid artery and a via right
jugular vein cannulation up to the right atrium for recording hemodynamic parameters and executing mechanical atrial stretch,
respectively. (Pesquisa Mdica 2:11, 2008). After 24-h, we continuously monitored the mean arterial pressure (MAP), central
venous pressure (CVP), heart rate (HR), as well as the cardiac output (CO) of all the rats which were subsequently distributed
into two groups randomly: rats submitted to mechanical stretching of the right atrium with a 50L intra atrial balloon over 5-min
(AS) or with a deflated balloon (sham stretch) until the end of study. In a group of rats, we studied the effect of AS on GR, by
feeding the animals with a test meal (1.5 mL, 0.5mg.ml-1 of phenol red in 5% glucose solution) 15 min after initial monitoring.
After a 10 min post-prandial interval, the rats were sacrificed by a thiopental overdose for the determination of GR using the dye
dilution retention method (Adv Physiol Educ. 33: 343, 2009). We determined OT plasmatic levels by radioimmunoassay in AS,
as well as sham stretch groups. In another set of rats, we verified the effect of AS on baroreflex responsiveness where after one
minute of AS, we administrated Phenylefrine (FE, 10 ug.Kg-1) or Sodium Nitroprusside (SNP, 10 mg.Kg-1) AND subsequently
analyzed the HR/MAP index. Data (mean SE) were analyzed using ANOVA followed by Student Newman-Keuls test,
considering values with p < 0.05 significant. When compared to the sham stretch group, AS promoted the increase of GR (54.1
3.0 vs 69.9 3.5%), CVP (1.2 0.5 vs 5.0 1.0 cmH2O), plasmatic levels of OT (3.6 0.5 vs. 6.8 0.5 pg.mL-1), as well as
tachycardia (371 7 vs. 421 22 b.p.m), but neither MAP (118.3 3.0 vs. 119.9 4.0 mmHg) nor values of CO (136.0 6.8 vs.
151.6 16.4 mL.min-1). Furthermore, we did not note significant alterations in the HR/MAP index after FE (-5.0 0.4 vs. 5.2 0.4 beats.min-1.mmHg-1) neither after prior SNP treatment (-2.20.4 vs. -2.20.5 beats.min-1.mmHg-1). However, right
atrium appendectomy prevented the effects induce by AS on GR (52.5 5.2 vs. 53.3 4.6 %), on CVP levels (12.4 3.2 vs. 14.2
3.0 cmH2O), OT plasmatic secretion (3.8 0.7 vs. 4.4 0.8 pg.mL-1) and on the reflexive tachycardia (475 14 vs. 501 6
b.p.m). Therefore, the MAP levels in this group remained within stable levels up to the end of studies (107.6 2.6 vs 109.4
3.1mmHg).
Conclusions:
The increases in gastric retention, as well as the hemodynamic and hormonal alterations due to right atrial stretch are mediated by
a neuroendocrine mechanism dependent on the integrity of the right atrial appendix, but not on the responsiveness of arterial
baroreceptors.
Keywords: ATRIAL STRETCH , HEMODYNAMIC, ENDOCRINE, GASTRINTESTINAL ACTIVITY , OXCYTOCIN
Financial Support: CAPES, FUNCAP, FAPESP e CNPq.
Resumo:04-034
GASTROPROTECTIVE ACTIVITY OF RIPARIN II ON ETHANOL-INDUCED GASTRIC LESIONS IN MICE:
INVESTIGATION OF POSSIBLE MECHANISMS OF ACTION
Feitosa, M. L. 1; Vasconcelos, L. F. 1; Ocarvalh, A. M. R. D. 1; Rocha, N. F. M. 1; Dias, M. L. 1; Fernandes,

M. L. 1; Silva, M. I. G. 1; Filho, J. M. B. 2; Gutierrez, S. J. C. 3; Sousa, F. C. F. D. 1
1
Fisiologia e Farmacologia/Faculdade de Medicina, UFC
2
Laboratrio de Tecnologia Farmacutica. , UFPB
3
Departamento de Bioquimica e Farmacologia / Farmcia, UFPI
Objectives:
The gastric injury by ethanol is a model widely used to experimental evaluation of antiulcerogenic drugs. This model has
fundamental importance for scientific research concerning the fact that we can evaluate possible mechanisms by which
substances may act promoting gastroprotection. In this study, we decide to evaluate the gastroprotective effect of riparin II
against ethanol-induced lesions and verify the role of nitric oxide, ATP-dependent K+channel and prostaglandins in this action.
Male Swiss mice (20-30g) were divided in four groups: vehicle (3% Tween 80 in distilled water; p.o.), ripII-25 (riparin II 25
mg/kg; p.o.), ripII-50 (riparin II 50 mg/kg; p.o) and CYP (Cyproheptadine 10 mg/kg; p.o.), used as a gastroprotective reference
drug. One hour later, absolute ethanol (0.2 mL/animal) was administrated orally to all groups. Thirty minutes after the
administration of ethanol, the mice were killed and their stomachs were removed and opened along the greater curvature for
examination. The total and injured stomach areas were measured by computer program and expressed in terms of percent (%) of
ulcerated gastric area. To study the possible mechanism of action, separate experiments were conducted using the following
drugs: L-NAME, an inhibitor of the NO synthase activity (10 mg/kg; i.p.), glibenclamide, an antagonist of KATP+ channels (10
mg/kg; i.p.) and indomethacin, a nonselective cyclooxygenase inhibitor (10 mg/kg; p.o.). L-NAME and glibenclamide were
administered 30 min before animals receiving riparin II (50 mg/kg; p.o.), while indomethacin (10 mg/kg; p.o.) was administrated
2 h before the drug test. One hour later, absolute ethanol 0.2 mL was applied in each animal. Thirty minutes after the
administration of ethanol, the mice were killed and their stomachs were removed for examination as previously described. Data
are presented by mean S.E.M and analyzed by ANOVA followed by Student Newman Keuls as the post hoc test. RESULTS:
The administration of absolute ethanol produced lesions in the gastric mucosa (17.840.482), which were reduced in the animals
pretreated with ripII-25 mg/kg (8.6750.816; p
Conclusions:
Our study conclude that riparin II has an gastroprotective effect and this effect appears to be involved, at least in part, with
endogenous NO and KATP+ channels opening.
Keywords: ulcer, ethaol, riparin II
Financial Support: CNPq/ CAPES
Resumo:04-035
EVALUATION OF GASTROPROTECTIVE ACTIVITY OF CHLOROGENIC ACID ON ETHANOL/HCL-INDUCED
ULCER MODEL
Shimoyama, A. T. ; Santin, J. R. ; Machado, I. D. ; Mller, G. G. ; Farsky, S. H. P.

Faculdade de Cincias Farmacuticas/USP, FCF/USP
Objectives:
The pathophysiology of gastric ulcer has focused on an imbalance between aggressive and protective factors in the stomach, such
as acid secretion, pepsin, mucus secretion, blood flow, cellular regeneration, prostaglandins and epidermal growth factors.
Chlorogenic acid (CGA) is a class of substances contained in a vast amount of plants, food and beverages. It's known to have
antioxidant effects, anti-inflammatory, hypotensive and anti-diabetic, but the literature lacks information regarding this acid to be
effective as a gastroprotective agent. Thus, the aim of the present study was to evaluate the activity of CGA using ethanol/HClinduced ulcer model in mice.
Swiss male mice (n=5) were orally treated with CGA (5, 25 or 50 mg/kg), Omeprazole (30 mg/kg) or vehicle (control). One hour
later, gastric ulcer was induced by administration of ethanol/HCl (1.0mL of ethanol 60% + HCl 0.03M, gavage). One hour after
the ulcer induction, animals were anesthetized and sacrificed, and gastric ulcers were quantified by measuring the percentage of
the injured area. Nitric oxide (NO) was measured in stomach homogenate, by Griess reaction. CGA or omeprazole treatments
significantly decreased (p
Conclusions:
Results obtained suggest that CGA may be a gastroprotective agent in the ethanol/HCl-induced ulcer model, which supports
further experiments to investigate the mechanisms involved in this protective action.
Keywords: Chlorogenic Acid, Ethanol, Gastroprotective, Ulcer
Financial Support: FAPESP (2010/18069-9)
Resumo:04-036
EFFECTS OF PEROXISOME PROLIFERATOR ACTIVATED RECEPTOR GAMMA (PPARGAMMA) PARTIAL
AGONIST LYSO-07 ON ETHANOL/HCL-INDUCED ULCER MODEL.
Santin, J. R. 1; Machado, I. D. 1; Shimoyama, A. T. 1; Mller, G. G. 1; Galdino, S. L. 2; Pitta, I. R. 2; Farsky,

S. H. P. 1
1
Faculdade de Cincias Farmacuticas, FCF/USP
2
Objectives:
Gastric ulcer is characterized by lesion formation and gastric tissue necrosis. This process is also marked by intense neutrophil
migration which contributes to the damaging process. Recently, it has been proposed that of peroxisome proliferator activated
receptor (PPARs) agonists can be used as anti-ulcer agents. Thus, this study aimed to evaluate the effects of PPAR partial
agonist Lyso-07 in ethanol/HCl-induced ulcer model.
Swiss male mice (n=5) were orally treated with Lyso-07 (5, 25 or 50 mg/kg), pioglitazone (40 mg/kg, control PPAR),
omeprazole (30 mg/kg) or vehicle (control) one hour before inducing gastric ulcer by oral administration of the damaging agent
(0.5mL of ethanol 60% + HCl 0.03M). One hour later, animals were anesthetized, blood was collected by puncturing the
abdominal aorta and the femurs exudates were removed for total leukocyte counting, using a Neubauer chamber. The subtypes of
leukocytes in the blood and in the bone marrow were quantified in May-Grunwald stained blood smears using optical microscope
and by flow cytometry, respectively. The stomachs were removed to measure the percentage of the injured area. Lyso-07
treatments (25 or 50 mg/kg) significantly decreased (p
Conclusions:
Data herein obtained suggest that the Lyso-07 presented gastroprotective activity in the ethanol/HCl-induced ulcer. In addition,
Lyso-07 affect the neutrophil traffic from bone marrow into blood, which may contribute to the gastroprotective effect, inhibiting
the migration of inflammatory cells into the injured tissue.
Keywords: gastric ulcer, PPAR , neutrophil, ethanol, gastroprotective
Financial Support: Fapesp (2010/17175-0), CNPq and Capes.
Resumo:05-021
IDENTIFICATION AND CYTOCHEMISTRY ANALYSIS OF NUCLEAR ANNULUS OF BOVINE SPERM
Lemos, M. S. ; Moreira, P. F. S. ; Moraes, A. S. ; Beletti, M. E.

rea de Cincias Morfolgicas-Univ. Federal de Uberlandia, UFU
Objectives:
The objective of this study was to isolate and identify the biochemical nature of the nuclear structure of the annulus, a component
of the nuclear matrix of bovine spermatozoa.
Semen samples were washed with PBS and subjected to ultracentrifugation in the concentration gradient of cesium chloride and
sucrose. The pellet obtained was treated with Triton X-100 at 10% for 24 hours and then treated with a solution of DTT
(dithiothreitol) 40 mM for 24 hours. After all steps several glass slides have been prepared for viewing by scanning electron
microscopy and for the testing cytochemistry. Three cytochemical stains were done: "Xylidine ponceau (XP) for protein, PAS
(periodic acid-Schiff staining) for neutral sugars and glycoproteins, and toluidine blue pH 4.0 for DNA. The first staining was
done with 0.1% XP solution placed on the blade by 15minutes; the PAS staining was performed with 0.5% periodic acid for 10
minutes and Schiff's reagent for 25 minutes and the third staining was done with 0.025% toluidine blue pH 4.0 for 15minutes.
After staining, the slides were examined under a light microscope coupled with a system for image capturing. Using the scanning
electron microscope, the nuclear annulus appeared as an elliptical ring with a membrane partially or completely occluding the
interior. Cytochemistrically the annulus was positive for XP and PAS staining, demonstrating to have a glycoprotein constitution.
The toluidine blue staining showed an elliptical ring with long filaments attached, demonstrating that DNA strands are firmly
adhered to the nuclear annulus.
Conclusions:
The study allowed the cytochemical characterization of nuclear annulus structure as a glycoprotein complex with attached DNA.
Keywords: nuclear annulus, bovine sperm, cytochemical analysis
Financial Support: CNPq and FAPEMIG
Resumo:05-022
COULD THE MATERNAL EXPOSURE TO ANTIDEPRESSIVE FLUOXETINE ALTER THE FERTILITY IN MALE
RAT OFFSPRINGS?
Vieira, M. L. ; Hamada, R. Y. ; Ferreira, P. C. L. ; Barbieri, M. ; Mesquita, S. F. P. ; Gerardin, D. C. C.

UNIVERSIDADE ESTADUAL DE LONDRINA, UEL
Objectives:
Evidence suggests that maternal depression during pregnancy is at least as common as postpartum depression. The use of
psychotropic drugs during these periods is in most cases essential. Fluoxetine (FLX), a selective serotonin (5HT) reuptake
inhibitor drug, has been widely prescribed for depression during pregnancy and lactation. 5HT hypothalamic concentration is
reduced in critical periods of brain masculinization on male rats, this probably happens to allow adequate testosterone action. In
this sense, levels of 5HT changes could influence the hypothalamic sexual differentiation that occurs in the perinatal period. It is
known that 5HT antagonizes testosterone masculinizing effects on size of sexually dimorphic hypothalamic nuclei and
gonadotropin release. In this way, the use of FLX by mothers could disrupt brain development resulting in reproductive
alterations in their progeny. The objective was to investigate possible changes on male offspring through reproductive
parameters, resulting from maternal exposure to FLX.
Wistar rats received FLX 7.5 mgkg or saline solution (CT) 0.9% (by oral gavage) from gestational day one to postnatal day
(PND) 21. On PND 90, the following parameters were observed in male pups: body and reproductive organs (testis and
epididymis) weights, sperm counts in the testis and epididymis. The other testis was submitted to the usual histological routine
for analysis in light microscopy and morphometric analyses were realized. Data SEM were compared by Students t test with
Welchs correction (p0.05).
Conclusions:
On basis of above observations, the results suggest that maternal exposure to FLX may have diminished endogenous testosterone
in male pups, which is crucial for brain sexual differentiation. This reduction has probably led to alteration on the
hypothalamuspituitarygonad axis in adulthood.
Keywords: ANTIDEPRESSIVE, ENDOGENOUS TESTOSTERONE, FERTILITY, MALE RAT, MATERNAL EXPOSURE
Financial Support: PIBIC/UEL.
Resumo:05-023
RAT SPERMATOGENESIS IN VITRO: GERM CELL DIFFERENTIATION IN A CO-CULTURE MODEL.
Pimenta, M. T. ; Jasiulionis, M. G. ; Porto, C. S. ; Lazari, M. F. M.

Department of Pharmacology - Experimental Endocrinology, UNIFESP - EPM
Objectives:
Spermatogenesis is a complex process that involves the interaction of germ and somatic cells. Mechanisms mediated by Sertoli
cells can regulate the proliferation, differentiation, and development of germ cells. The in vitro production of meiotic and
postmeiotic germ cells would indicate an optimal culture condition for survival and differentiation. However in vitro systems for
the study of mammalian spermatogenesis have been severely limited since co-cultures of testicular cells fail to allow
differentiation of germ cells. The aim of this study was to establish and characterize a new co-culture model to improve the
proliferation and differentiation of germ cells.
All experimental procedures were approved by the Ethical Committee from UNIFESP (CEP 0937/10). Primary co-culture was
obtained from 7-day old male Wistar rats, whose testes have higher percentage of gonocytes and type A spermatogonia. Testes
were removed, decapsulated and subjected to three-step enzymatic digestion under shaking conditions: the first using trypsin and
DNase; the second using glycine and DNase and the third using collagenase and DNase. The cells were cultured on Matrigel
(Becton Dickinson) covered dishes, in DMEM/F12 (phenol red-free) containing 0.02 g/L of gentamicin, for different periods (4,
7, 10 and 15 days), at 35C, 5% CO2. After several attempts, the best supplementation condition was obtained using 0.5% fetal
bovine serum (FBS). Proliferation and differentiation of the cells was analyzed by several methodologies: 1) histochemistry and
morphological analysis by optical microscopy; 2) immunocytochemistry using markers for undifferentiated (PLZF,
promyelocytic leukemia zinc-finger) and differentiated (c-KIT) germ cells; 3) Real time PCR with Sybr Green reagent (Applied
Biosystems) to determine the mRNA levels for specific markers of cell proliferation and differentiation (proliferating cell nuclear
antigen, Pcna, and histone H1-like protein in spermatids 1, Hils1); and 4) flow citometry using nucleic acid staining with
propidium iodide (PI) to evaluate cell cycle and the cell ploidy. Germ cells expressing PLZF and c-KIT could still be detected
after 7 days of culture, but morphological analysis revealed the presence of elongated spermatids already by culture day 5. The
quantitative PCR confirmed the increase of Pcna and Hils1 transcripts at specific stages during culture. Flow cytometry allowed
identification of three populations of cells, diploid, tetraploid and haploid, and the influence of medium supplementation on the
relative proportion of these populations. It was noticeable the appearance of additional hypofluorescent haploid peaks during
culture, reflecting different degrees of chromatin condensation.
Conclusions:
We developed a co-culture system of Sertoli and germ cells with appropriate conditions to allow cell proliferation and
differentiation. This model can now provide an important tool to investigate critical points in the regulation of proliferation and
differentiation of germ cells.
Keywords: co-culture, differentiation, germ cell, proliferation, spermatogenesis
Financial Support: FAPESP, CAPES, CNPq.
Resumo:05-024
RESIDUAL OIL FLY ASH (ROFA) EXPOSURE PROMOTES TESTICULAR OXIDATIVE STRESS IN WISTAR
RATS
Motta, G. A. ; Halmenschlager, G. ; Damiani, R. M. ; Piva, M. O. ; Rhoden, C. R. ; Rhoden, E. L.

Universidade Federal de Cincias da Sade de Porto Alegre, UFCSPA
Objectives:
Epidemiologic studies have demonstrated increased human morbidity and mortality due to elevations in the concentration of
ambient air particulate matter (PM) (N Engl J Med 343:17421749, 2000). In addition, recent studies have shown that PM can
lead to male reproductive toxicity (Inhal Toxicol 19:275281, 2007). Therefore, the aim of this study was to analyze the response
of testicular tissue of rats after a chronic exposure to different concentrations of ROFA.
Methods: A total of 35 male Wistar rats were divided into four distinct groups: G1 (Saline, 10L; n=8), G2 (ROFA 50g/10L,
n=9), G3 (ROFA 250g/10L; n=9) and G4 (ROFA 500g/10L; n=9). They were exposed to ROFA during 90 days by
intranasal instillation. After this period, the animals were euthanized and the testicles were removed and frozen at -80C.
Oxidative analyses were performed on testicles through the Thiobarbituric Acid Reactive Substances test (TBARS), which
measures malondialdehyde (MDA) concentration, and through the activity of the antioxidant enzymes catalase (CAT) and
Superoxide Dismutase (SOD). Data are demonstrated as mean standard deviation and were statistically analyzed by one-way
ANOVA, followed by Tukey post-hoc test (P
Conclusions:
The exposure of testicular tissue under different doses of ROFA promoted an increase of SOD activity, the first line of
antioxidant defense, but decreased CAT activity, causing testicular lipid peroxidation.
Keywords: exposure, oxidative stress, ROFA, testicles
Financial Support: CAPES, CNPq and UFCSPA
Resumo:05-025
TRIBUTILTIN AS A POTENTIAL RAT REPRODUCTIVE CICLE DISRUPTOR
Filho, V. S. D. 1; Lopes, P. F. I. 1; Podratz, P. L. 1; Costa, M. B. D. 2; Matsumoto, S. T. 2; Samoto, V. Y. 3;

Silva, I. V. 1; Takiya, C. M. 3; Graceli, J. B. 1
3
INSTITUTO DE CINCIAS BIOMDICAS/UFRJ, UFRJ
1
DEPARTAMENTO DE MORFOLOGIA/UFES, UFES
2
DEPARTAMENTO DE BIOLOGIA/UFES, UFES
Objectives:
Organotin compounds (OT), such as tributiltin (TBT), have been widely used as biocides and disinfecting agents in circulating
industrial cooling waters, as well as antifouling paints for marine vessels. There are many reports of the biological effects of OT,
which vary in their toxic effects on eukaryotes. These OT are potent endocrine disrupters in gastropods, through the inhibition of
aromatase, changed reproductive parameters, and induced mollusks sterile by the development of a syndrome called imposex.
The importance of OT as environmental endocrine disrupters and their potential to adversely affect human health, was previously
demonstrated in several comprehensive studies of the reproductive toxicity of TBT in rodent models, however the toxic effects in
reproductive cycle not much is understood. We evaluated the potential toxicity of TBT as reproductive disruptor in female rats, as
impairer of regular reproductive cycling correlated with changes in sexual steroids.
Wistar female rats presenting normal estrus cycle (230g, aged 3 months, R. norgegicus) were randomly divided in: 1) Control
(CON, treated with vehicle by gavage, n=8); 2) Treated with TBT (TBT, 100ng/Kg, treated with TBT by gavage, n=8) . After 20
days of daily treatment, we analyzed the estrous cycle (EC), percentage (%) of cycling, phases of the estrous cycle, serum level of
17-estraiol (E2), and progesterone (P4), weight of uterus, weight of ovaries, comet and micronucleus analysis in erythrocytes
and, micronucleus analysis in ovarian cells of hamster (CHO-K1) line. Also, the uterine and ovarian tissues were analyzed with
histological sections stained with hematoxylin and eosin. Results are presented as meanSEM. One-way analysis of variance
(ANOVA) followed by unpaired Student t-test). Differences were assumed to be significant when p
Conclusions:
These data suggest that serum changes in female sexual hormones levels, induced by TBT , impairs cyclic reproductive function,
causes morphofunctional damage in reproductive organs and possesses a toxic potencial over female rats reproductive system.
Keywords: endocrine disrupter, estrous cycle, female reproductive organs, triorganotins
Financial Support: UFES e FAPES (n 45446121/09)
Resumo:05-026
QUALITATIVE VALIDATION OF AN ARTISAN HOUSE FOR EXPOSURE OF RATS PARTICULATE MATTER.
Campos Junior, O. ; Dias, F. C. R. ; Melo, T. N. T. ; Neves, E. M .

departamento anatomia/universidade federal de pernambuco, UFPE
Objectives:
Several studies have linked the continued exposure to ambient levels of particulate matter (PM) and reduced life expectancy and
increased mortality and morbidity. The objective of this study was to develop and validate a craftsmanship exposure chamber,
where rats were exposed to dust from diesel (MP) via nebulization.
The analysis of the temperature, humidity and pressure of the chamber of handicraft exhibition were taken using a digital thermohygrometer and a monomer of 60psi/4Kgf/cm2. The powder used for diesel exposure, kindly provided by the Laboratory of
Experimental Air Pollution, University of So Paulo (IPL / USP), was diluted in distilled water at three different concentrations:
1g, 10 mg and 50 mg. They used eight rats Rattus norvegicus albinos with 90 days of age were divided into four groups (n = 2):
Control Group (CG), Group 1 (G1) and Group 10 (G10) and Group 50 (G50), respectively exposed nebulization of 3 ml of
distilled water (GC), 1g (G1), 10 mg (G10), 50 mg (G50) of diesel dust suspended in 3ml of distilled water. The four
experimental groups were subjected to the nebulizer for 1 hour daily for a period of 8 consecutive days. The animals were
weighed before exposure and 1 second after the last exposure (8th exposure). After the last day of exposure and weighing, the
animals were anesthetized (ketamine and xylazine) and lungs were collected and fixed by immersion in Bouin fixative. After
routine histological processing, the slides were examined in bright field microscope. The exposure chamber had a craft outlet
pressure of 5psi, the same business model presented by INALTEC PLUS 6600, which has an air flow of 8L/min and generation
of aerosols in the range 0.3 to 0.8 micrometers. Qualitative assessment of lung histology revealed an inflammatory process in
early G1, progressing to pictures of emphysema and hyperplasia in groups G10 and G50, respectively, with the presence of
particulate matter in the cluster lung parenchyma and macrophages.
Conclusions:
The qualitative assessment of the lungs of animals exposed to dust from diesel exposure chamber validates the craft as a system
of exposure to particulate matter.
Keywords: pollution , particulate matter, inhalation
Financial Support: National Institute for Integrated Analysis of Environmental Risk - INAIRA
Resumo:05-027
EFFECT OF AGENTS FOUND IN AIRPOLLUITION OF THE CITY OF RECIFE-PE ON THE IGS WISTAR RATS.
Melo, T. N. T. ; Dias, F. C. R. ; Campos Jr. O. . ; Lima Jr. M. J. M; Neves, E. M.

departamento anatomia/universidade federal de pernambuco, UFPE
Objectives:
With growing concern about the adverse effects of air pollution on health, developed a project that aims to find out if air pollution
is affecting the city of Recife the testicle, and thereby compromising the reproduction. In this context, we present biometric
parameters testis of adult rats subjected to air pollution.
Methods: Wistar male rats formed three groups: polluted F1 (F1, n = 10); polluted group F2 (F1, n = 13) and control (C, n = 6)
consisting of mice generated, breastfed and kept in the vivarium until euthanasia. Animals in groups F1 and F2 were kept in a
room of pollution (area = 4.8 m2; 14.500L volume) with constant flow of air polluted city of Recife, directed by a conduit with
PVC 100mm diameter. Rats with F1 were generated and maintained in room breastfed pollution with two objectives: 1)
Formation of the F1 group (n = 10), and 2) Playing for the formation of F2 (n = 13). All experimental animals received water and
food ad libitum and were subjected to a reversed cycle of daily 12h light and 12h dark, temperature between 20-240C, with
humidity between 45-75% and air exhaust system installed. At 90 days old mice of experimental groups (F1, F2 and C) were
weighed, anesthetized, fixed with Bouin, orchiectomized, and testes were weighed. After obtaining the body and testicular
weights, we calculated the gonadosomatic index (GSI): ratio between the testicular mass and body weight for all rats of three
experimental groups (F1, F2 and C).Statistical analysis used the Student t test. The level of statistical significance was considered
p 0.05. ts: The average body weights obtained at the end of exposure showed no statistical difference between experimental
groups (365 44g = F1, F2 = 354 30g, C = 369 16g). The average testis weight was significantly greater in the F2 group,
when compared with groups C and F1 (F1 = 1.68 0.09 g; GF2 = 1.78 0.09 g C = 1.65 0.10 g) .The GSI Group F2 was
significantly higher compared to the IGS group C (C = 0.89 0.05%, F1 = 0.93 0.11%, F2 = 1.01 0.076%).
Conclusions:
In Recife, the average annual PM2, 5 was 11g / m, staying within the estimate by the WHO (25g / m) (17. International
Symposium on Undergraduate Research, University of So Paulo, 844, 2009), but through these results we suggest that the level
of air pollution in the city of Recife was enough to promote change in testis size, especially in the F2, F1 whose parents had been
created under the effect of air pollution.

Keywords: polluition, particulete matter, spermatogenesis
Financial Support: National Institute for Integrated Analysis of Environmental Risk - INAIRA
Resumo:05-028
EFFECT OF CHRONIC MALNUTRITION IN RAT VAS DEFERENS: MORPHOLOGYCAL AND BIOCHEMICAL
ANALYSIS
Bezerra, C. G. P. 1; Souza, A. M. 1; Muzi-filho, H. 1; Boldrini, L. C. 2; Takiya, C. M. 2; Nesi, R. T. 2;

Valena, S. S. 1; Einicker-lamas, M. 3; Vieyra, A. 3; Lara, L. S. 1; Cunha, V. M. N. 1
1
Programa de Farmacologia Celular e Molecular - ICB, UFRJ
2
Programa de Cincias Morfolgicas - ICB, UFRJ
3
Objectives:
Malnutrition observed during pregnancy and the first days of life promotes adaptive changes related to a higher risk of disease in
adulthood. In this context, we previously demonstrated changes in calcium-dependent processes of rat vas deferens (RVD) that
seems to compromise male reproductive capacity. The aims of the present work were: 1-investigate the RVD architecture; 2evaluate the Ca2+ sensitivity of the Ca2+-ATPases from different types (SERCA and PMCA); 3- investigate if protein kinase A
(PKA) or protein kinase C (PKC) can phosphorylate the Ca2+-ATPases; 4- evaluate a possible oxidative damage caused by
malnutrition.
Animals were chronically malnourished from weaning until 13 weeks of age (CM group) using the model described as Regional
Basic Diet (Arch. Latinoam. Nutr. 40; 533, 1990). After this period, control and CM male rats were sacrificed (CEUA
DFCBICB007). Histomorphometric analysis were performed according to Masson's trichrome method, in which atrophy was
observed in the prostatic portion of male rats from CM group (59% of control muscle mass; n=3). The measurement of enzymatic
hydrolysis of gama-32P[ATP] showed that Ca2+-ATPases present in the RVD homogenate exhibited an increased of Vmax
values 653% (SERCA) and 603% (PMCA), n=5 and a reduced Km value for SERCA (27%; n=5) in CM group. The
phosphorylation of the Ca2+-ATPases of different types was seen in a specific film (phosphor screen) after the 32P-proteins be
separated by SDS-PAGE electrophoresis. Chronic malnutrition promotes increase in the PKAi-sensitive phosphorylation (related
to PKA) of the 140 and 110 kDa bands, related to PMCA (487%; n=3) and SERCA (186%; n=3), respectively. By the other hand,
an increase in the calphostin C-sensitive phosphorylation (related to PKC) was detected only in the 140 kDa band, corresponding
to PMCA (186%; n=3). In CM group, it was also observed a significant increase of lipid peroxidation by TBARS method (659%;
n=3) as well as protein carbonylation (292%; n=6).
Conclusions:
Taken together, the present data help to understand the Ca2+-dependent molecular events that affect reproductive capacity of
adult male rats chronically malnourished. It seems that the oxidative damage of membranes and protein constituents in vas
deferens cells alters calcium homeostasis. In this context, the Ca2+-ATPases show high hydrolytic activity and elevated levels of
phosphorylation. However, it seems that all of these efforts (and others) are not enough to reestablish the normal contractility of
vas deferens as it is observed muscle atrophy, and as previously reported, it be may related to a deficient transport and raise of
death of sperm cells observed in this organ.

Keywords: Rat vas deferens, chronic malnutrition, regulatory phosphorylation, oxidative damage, kinetics of calcium pumps
Financial Support: Projeto Casadinho-CNPq; PROCAD-CAPES; FAPERJ Primeiros Projetos, Programa

ALV
Resumo:05-029
EFFECTS OF CHRONIC NITRIC OXIDE SYNTASE INHIBITION ON RAT PROSTATE SMOOTH MUSCLE
REACTIVITY.
Calmasini, F. B. ; Leiria, L. O. S. ; Pissinatti, L. ; Antunes, E.

Department of Pharmacology / Faculty of Medical Sciences , UNICAMP
Objectives:
The prostate is a gland innervated mainly by adrenergic and cholinergic fibers that act synergistically controlling the smooth
muscle reactivity. Non-adrenergic non-cholinergic (NANC) purinergic fibers, through the release of adenosine triphosphate
(ATP), also play a role in the smooth muscle contraction tone regulation. Moreover, signal transduction pathways using NOcGMP regulates diverse physiological functions such as smooth muscle contractility, neurotransmission and cell
growth/proliferation. However, the NO-cGMP signaling system in prostate smooth muscle regulation is still poorly investigated.
Chronic NO inhibition in different tissues lead to functional, molecular and biochemical alterations, and therefore has been
widely used as a pharmacological strategy to study the role of NO in several tissues. Therefore, the aim of this study was to
evaluate the effects of chronic NO blockade in the rat isolated prostate smooth muscle.
The experimental protocols were approved by the Animal Ethical Committee of UNICAMP. Male Wistar rats were treated or not
with the NO synthesis inhibitor L-NAME (20 mg/rat/day) given in the drinking water for 4 weeks. After treatment, animals were
sacrificed through isoflurane overdose, and the prostate was isolated and weighed. Concentration-response curves to
phenylephine (PE, 1 nM-100 M), carbachol, (CCh, 1 nM - 100 M), and ,-methylene ATP (1, 3 and 10 M) were obtained.
The neurogenic contractile responses induced by electrical-field stimulation (EFS, 1-32Hz) were also obtained. Values of potency
(pEC50) and maximal responses (Emax) were calculated. Histological analyses in prostates from control and L-NAME-treated
rats were performed. Chronic L-NAME administration promoted a significant increase the prostate weight (503 27.6 mg, n=5, p
Conclusions:
Our findings show that long-term NO deficiency in rats causes in vitro functional alterations characterized by increased
contractile responses to -adrenoceptor and muscarinic receptor activation, but not to P2X1 activation. Whether chronic NOdeficiency in rats mimics the benign prostatic hyperplasia is under investigation.
Keywords: prostate, L-NAME, nitric oxide
Resumo:05-030
COMPARATIVE STEREOLOGICAL STUDY ON THE MORPHOLOGY OF THE SEMINIFEROUS TUBULES OF
RATS FED CANOLA AND SOYBEAN OIL DIETS.
Almeida, A. F. G1; Costa, C. A. S. 3; Nascimento - Saba, C. C. A. 3; Rueff-barroso, C. R. 1; Santos, R. M.

M. 2; Lancetta, C. F. F. 1
1
Departamento de morfologia , UFF
2
Departamento de fisiologia e farmacologia, UFF
3
Departamento de cincias fisiolgicas, UERJ
Objectives:
The intake of polyunsaturated fatty acids (PUFAs), especially omega 3 (n-3) and omega 6 (n-6), is essential to promote the proper
development and maintenance of various physiological systems. In Brazil, the soybean oil (composed of 8% of n-3 and 54% of n6) is the most consumed oil in all social classes, corresponding to an average of 82% of total calories obtained from vegetable oils
and fats, while canola oil (comprising 11% of n-3 and 21% of n-6) is not much consumed, corresponding to an average of 4.2%
(Rev Sade Pblica. 2005 Aug;39(4):530-40). The human body has a preference to metabolize n-3 fatty acids, and it is believed
that the ideal proportional intake of n-3 and n-6 is 5 molecules of n-6 for 1 molecule of n-3. However, there are reports in which
the proportion vary from 3:1 to 10:1. Based on this information, the purpose of this study was to evaluate the best ratio of intake
of these fatty acids, comparing diets with soybean and canola oils, on the developmental of seminiferous tubules.
10 male Wistar rats were divided into two groups: 5 animals were fed diets containing 7% soybean oil (control group) and 5
others received diets containing 7% canola oil (canola group). At 60 days these animals were sacrificed and testicles processed
following routine techniques for paraffin embedding. The volume density (Vv), the length density (Lv), surface density (Sv) and
the transverse area of testicular tubules were determined by stereological methods. Data were analyzed using t test considering p
Conclusions:
Animals fed canola oil had lower testicular growth than those fed with soybean oil, showing that the highest proportions of n-6
compared to n-3, present in soybean oil, are probably more beneficial to testicular development.
Keywords: canola oil, Essential fatty acids, seminiferous tubules, soybean oil, testicle
Resumo:07-011
EFFECT OF UREA ON THE ACTIVITY OF ABCC1 AND CELL VIABILITY OF THE MA104 CELL LINE
Santos, D. H. F. 1; Fonseca, L. M. 2; Capella, M. A M. 1,2; Lopes, A. G. 1

1
Instituto de Biofsica carlos Chagas Filho, IBCCF
2
Instituto de Bioqumica Mdica, IBqM
Objectives:
The accumulation of urea in the renal medulla is due to the action of easily regulated transporters and is a critical factor for the
mechanism of urinary concentration. Its known that high concentrations of urea activate signaling pathways in kidney epithelial
cells. Urea itself is a known denaturant agent, but kidney cells are capable of surviving in the presence of high urea
concentrations.Previous data from our group showed that Ma104 cells are resistant to urea and that this molecule increased the
viability of NaCl treated cells. We have also shown that both NaCl and urea are capable of modulating the ATPase activity of
ABCC1. In this work we aim to evaluate the cellular responses of Ma104 cell line to hyperosmolar conditions due to urea.
Cells were incubated in transwell plates (5x105 cells/ well) for Five days until the monolayer was completely established. After
that, 9 mg of urea were added to the upper medium, lower medium, both mediums, or none, creating three treatment conditions
and one control condition. Urea dosage was performed using a commercial kit based on urease activity after 6, 24 and 48 hours of
incubation. After 6 hours of incubation the values measured of urea in the three ocnditions were: Urea top: 5,3500,1919 mg,
Urea bottom: 2,2180,1640 mg. (P
Conclusions:
The results suggest that the treatment with urea does not affect the viability of both the Ma104 cells and the MDCK cell line. It
does, however reduce the transport activity of ABCC1, which might be linked to an attempt of preserving intracellular levels of
GSH, which is paramount to cell survival in hyperosmotic environments. Its also important to note that the Ma104 cells show a
bidirectional transport of urea, indicating the presence of urea transporters in both the basolateral and apical membranes.
Keywords: urea, Ma104, hyperosmolar, activity, viability
Financial Support: CAPES, CNPq, FAPERJ, Ary Frauzino Foundation (FAF-ONCO II)
Resumo:07-012
QUALITY OF EVOLUATION OF DIABETIC PATIENTS ON HEMOLIADILYSIS IN THE CITY OF PELOTAS/RS
2010, ACADEMIC WORK NURSING COLLEGE. FEDERAL UNIVERSITY OF PELOTAS, PELOTAS/RS.
Noschang, J. 2; Vidales-braz, B. M. 1; Carvalho. M. P. 3; Peres, W. 4; Guanilo, M. E. E. 1

1
Faculdade de Enfermagem, UFPEL
2
Faculdade de Medicina, UCPEL
3
Faculdade de Fisioterapia, UCPEL
4
Centro de Cincias Qumicas Farmacuticas e de Alimentos, UFPEL
Objectives:
The chronic renal failure nowadays represents an important public health problem in the world, being considered as an
epidemic of alarming growth. In Brazil, currently, it is estimated that there are over two million Brazilians bearing
some degree of renal dysfunction. The diabetic nephropathy (DN) is the main cause of chronic renal failure, 40% of DN
bearers need dialysis. Purpose: Evaluating the Quality of life related to health (QLRH) of diabetic patients with
nephropathy submitted to hemodialysis in two specialized centers in the city of Pelotas/RS.

Observational study of transversal type quantitative delineation and of descriptive character which used as collection
instruments a socio-demographic questionnaire and the Kidney Disease and Quality of Life Short Form (KDQOLSFTM), as it consists of a specific instrument to be applied in individuals with chronicle renal disease on some type of
dialysis program. The data obtained were processed and analyzed in the SPSS program - Statistical Package for the
Social Sciences - version 15.0. Results: The sample had a total of 35 patients, in which 23 (65,7%) were males. The general
age ranged from 38 to 78 years (M: 61,3; a DP: 13,8) and the time of hemodialysis varied from 1 to 144 months. The
dimensions which presented higher average rates were a stimulus from the team (82,12), quality of social interaction
(76,00) and list of symptoms and problems (75,89). The lower rates were for emotional function (28,57), physical function
(30,00), work situation (31,32) and burden of kidney disease (37,50).
Conclusions:
It is believed that this study has positively contributed for the identification of important matters concerning the health of
these patients, as well as for the implementation of new strategies to care for diabetic patients with nephropathy on
hemodialysis in the city of Pelotas/RS, as it provides scientific subsidies which indicate a committed QLRH.
Keywords: Diabetic, diabetic nephropathy, Evaluating the Quality of Life, hemodialysis
Resumo:07-013
PROGRESSION OF DIABETIC NEPHROPATHY IN RATS
Jdice, M. N. ; Castiglione, R. C. ; Barbosa, C. M. ; Sousa-menezes, J. ; Morales, M. M.

Objectives:
Diabetic nephropathy is one of the most serious complications of diabetes mellitus (DM) and the most common cause of endstage renal failure. The main feature of early functional renal changes in diabetes is glomerular hyperfiltration. In the present
study we investigated the temporal evolution of renal dysfunction in rats submitted to streptozotocin induced diabetes.
Diabetes was induced by a single dose of streptozotocin (45mg/Kg, i.p.) in 8 weeks-old male Wistar rats (180-200 g) (Protocol
approved by Animal Ethics Comittee). The animals were divided in two groups: controls (CTRL, n = 4-6) and diabetics (DM, n =
4-5). Glycemia was determined twice a week (blood tail), using the glucometer OneTouch Ultra2. DM animals remained
hyperglycemic until 28 weeks after diabetes induction (glycemia higher than 200 mg/dL). Glomerular filtration rate (GFR), water
and food ingestion, weight gain/loss, glycemic levels and renal excretion of total proteins, glucose, and albumin were analyzed in
4, 8, 12, 16, 20, 24 and 28 weeks after diabetes induction. For urine collection and analyses of food and water intake, the animals
were placed in metabolic cages for 24 hours and then were subjected to ketamine (75mg/Kg) and xylazine (10mg/Kg) anesthesia
(i.p.) for blood collection. DM animals did not show a normal increase in body weight, as observed for CTRL animals, despite
the higher ingestion of water and food observed in DM compared to CTRL animals. DM animals showed an increase in urinary
flow 4, 8, 12, 16, 20, 24 and 28 weeks after the diabetes induction (37.411.01, 50.284.69, 39.938.79, 46.796.14, 50.887.44,
28.838.21, 45.223.34 L/min respectively) compared to CTRL groups (7.761.33, 5.900.99, 6.951.16, 5.250.49,
5.662.05, 6.230.26, 4.830.52 L/min respectively). The same pattern of increase was observed in GFR in DM groups
(10.181.17, 5.131.09, 5.521.07, 9.902.90, 5.341.99, 4.38 0.96, 5.760.65L/min/g respectively) compared to CTRL
groups (5.300.59, 3.290.54, 1.710.48, 3.700.24, 0.290.11, 1.640.59, 1.4800.06 L/min/g, respectively). Proteinuria was
higher in DM groups 4, 8, 12, 16 and 20 weeks after diabetes induction (18.104.72, 10.452.70, 23.153.83, 26.132.92 and
44.824.03 mg/day, respectively) compared to CTRL groups (10.011.84, 5.181.07, 10.801.33, 10.270.86 and 13.791.02
mg/day respectively). Osmolar clearance was increased in DM groups 4, 8, 12, 16, 20, 24 and 28 weeks (0.070.01, 0.080.020,
0.10.032, 0.130.019, 0.220.021, 0.290.013, 0.330.01mL/min, respectively) compared to CTRL groups (0.0240.00017,
0.0250.002, 0.030.003, 0.0320.0017, 0.0320.0022, 0.0350.0011, 0,0340,002 mL/min, respectively). Statistical analysis
was performed by t-tests and One-Way ANOVA (Newman-Keuls post-test). Differences were considered significant when
p<0.05.
Conclusions:
GFR and urinary flow were higher in DM compared to CTRL animals showing a change in the renal function, consequently,
producing a higher excretion of protein in the urine of DM animal model at 4, 8, 12, 16 and 20 weeks after induction of diabetes.
Keywords: DIABETIC NEPHROPATHY , glomerular filtration rate, hyperglycemia, rats, streptozotocin
Financial Support: FAPERJ, CNPq, CAPES, FUNEMAC, PRONEX
Resumo:07-014
HISTOLOGICAL AND ULTRASTRUCTURAL ANALYSIS OF THE RENAL PELVIS IN NORMAL AND DIABETIC
WISTAR RATS.
Lima, V. M. 1; Chopard, R. P. 2
1
DEPTO. DE CIRURGIA/FACULDADE DE MED. VETERINRIA E ZOOTECNIA, FMVZ/USP
2
DEPTO. DE ANATOMIA/INSTITUTO DE CINCIAS BIOMDICAS, ICBIII/USP
Objectives:
This project aims to describe and analyze the collagen, elastic and smooth muscle fibers of the renal pelvis of Wistar rats by
analyzing structural changes in experimental groups compared with the control group.
It was used male adult Wistar rats (Rattus norvegicus albinos) divided into two different groups (A and B), normal Wistar rats
and diabetic rats. The rats in group B were induced diabetes by Alloxan dissolved in physiological solution of sodium chloride
0.9% (135mg Alloxan in 6ml Saline), was then injected 0.2 ml of solution per 100g of body weight. The region of the renal pelvis
that contains representation of fibers was collected and reduced into small fragments. The cuts obtained are stained by the
following methods: Iron hematoxylin (Verhoeff) for demonstration of elastic fibers; Resorcin fuchsin (Weigert) for
demonstration of elastic and elauninic fibers; Azan for disclosure of the component collagen and smooth muscle; Picrosirius to
observe the component collagen, specifically collagens type I and III. The results were analyzed according to each technique.
Verified by light microscopy, the group of normal rats showed a higher area of collagen type I, if compared to the diabetic group.
We also observed that the collagen type III are not present in the two groups. Normal rats have two layers of collagen in the
submucosa and another one in the adventitia, with muscle fibers between them. The elastic fibers are well organized in lamellae
in the submucosa and sparce in the adventitia. Already in diabetic rats the layers of collagen are continuous, but disorganized.
The elastic fibers are sparse in the adventitia and interrupted in the submucosa, without lamellae. Ultrastructural results
demonstrated that in animals without induction of diabetes smooth muscle cells that form the renal pelvis are evenly arranged,
with slightly rounded nuclei and the presence of organelles such as mitochondria is constant, but not in large quantity. In the
diabetic animals we noticed that the nuclei of muscle cells are elongated, cytoplasmic organelles such as endoplasmic reticulum
and others are arranged unordered, also observed a significant increase of mitochondria and the presence of vilosity facing the
light of the renal pelvis.

Conclusions:
From the methodology and results, we infer that diabetes causes structural and ultrastructural differences renal pelvis of rats,
mainly in the organization of collagen and elastic fibers and also by the considerable increase of mitochondria in these rats, which
can demonstrate that they have a more intense energy expenditure.
Keywords: RENAL PELVIS, WISTAR RATS, ELASTIC FIBERS, COLLAGEN FIBERS, MUSCLE FIBERS
Resumo:07-015
GLUCAGON-LIKE PEPTIDE-1 RECEPTOR ACTIVATION INCRESES GLOMERULAR FILTRATION RATE AND
PROXIMAL TUBULAR SODIUM REABSORPTION
Crajoinas, R. O. 1; Oricchio, F. T. 1; Pessoa, T. D. 2; Pacheco, B. P. M. 1; Lessa, L. M. A. 2; Malnic, G. 2;

Girardi, A. C. C. 1
1
Instituto do Corao, HC-FMUSP
2
Fisiologia e Biofsica, ICB-USP
Objectives:
Glucagon-like peptide 1 (GLP-1) is a gut incretin hormone considered a promising therapeutic agent for type 2 diabetes because
it stimulates beta cell proliferation and insulin secretion in a glucose-dependent manner. Cumulative evidence supports a role for
GLP-1 in modulating renal function; however the mechanisms by which GLP-1 induces diuresis and natriuresis have not been
completely established. This study aimed to define the cellular and molecular mechanisms mediating the renal effects of GLP-1.
GLP-1 (1 g/kgmin) was intravenously administered in rats for the period of 60 minutes. Continuous infusion of GLP-1 induced
an increase in the urinary excretion of cAMP when compared to controls (121 9 vs. 8 3 pmol/min, P = 0.003). GLP-1 infusedrats displayed increased urine flow, fractional excretion of sodium, potassium and bicarbonate compared to those rats which
received vehicle (1% BSA/saline). GLP-1-induced diuresis and natriuresis were also accompanied by increases in renal plasma
flow and glomerular filtration rate. Real time RT-PCR in microdissected rat nephron segments revealed that GLP-1 receptormRNA expression was restricted to glomerulus and proximal convoluted tubule. In rat renal proximal tubule, GLP-1 significantly
reduced NHE3-mediated bicarbonate reabsorption via a protein kinase A (PKA)-dependent mechanism. Reduced proximal
tubular bicarbonate flux rate was associated with a significant increase of NHE3 phosphorylation at the PKA consensus sites in
microvillus membrane vesicles.
Conclusions:
Taken together, these data suggest that GLP-1 has diuretic and natriuretic effects which are mediated by changes in renal
hemodynamics and by downregulation of NHE3 activity in the renal proximal tubule. Moreover, our findings support the view
that GLP-1-based agents may have a potential therapeutic use not only as antidiabetic drugs but also in hypertension and other
disorders of sodium retention.
Keywords: incretin, protein kinase A, proximal tubule, renal hemodynamics, sodium reabsorption
Resumo:07-016
WATER AND SODIUM TRANSPORT INHIBITED BY THE ANTIMALARIAL DRUG ARTESUNATE IN RAT IMCD
PERFUSED IN VITRO.
Yano, Y. ; Rouch, L. K. ; Magaldi, A. J.

Hospital das Clinicas da Faculdade de Medicina da USP, HCFMUSP-LIM 12
Objectives:
Artesunate (Art) is a new effective antimalarial drug but a reversible polyuria and natriuresis has been related with its use
(Campos S.B., Kidney Int 2001,59:1044). This study was designed to determine the effect of Art on water and Na+ absorption in
rat IMCD.
The osmotic water permeability (Pf, &mum./sec) and Na+ lumem-to-bath flux (Jlb pEqcm2/sec) were determined in normal rats
IMCD(n=16) isolated and perfused in vitro by the standart methods. Art (10-5M) added to the bath in Vasopressin(Vp) absence
decreased Pf from 4.0b1.2 to -18.3&plusmn1.4 (p
Conclusions:
In summary, Art in IMCD: 1) inhibits partially the basal Pf and the Vp-stimulated Pf by 50% and totally the Vp-stimulated Na+
Jlb flux ; 2) is blocked by Anti-V2, and by Indo, leading us to suppose that its action could be by PGE2 stimulation synthesis by
mean of Vp receptors, explaining, at least in part, the observed polyuria and natriuresis during the use of this new antimalarial
drug .
Keywords: Artesunate, Sodium Transport, Vasopressin, Water Transport
Financial Support: FFM, LIMHCFMUSP, FMUSP,FAPESP,CNPq
Resumo:07-017
THE MATERNAL AMPLIFIED PERINATAL ESSENTIAL FATTY ACID DEFICIENCY DISCLOSES RENAL
DISFUNCTION IN THE ADULT OFFSPRING
Ribeiro, V. S. ; Silva, A. R. ; Pereira, S. F. J. ; Castro-chaves, C. ; Paixao, A. D. O.

DEPARTAMENTO DE FISIOLOGIA E FARMACOLOGIA, UFPE
Objectives:
The maternal amplified perinatal period in an essential fatty acid deficient diet (EFAD) effects on renal function and blood
pressure in the adult offspring were investigated.
All rats were maintained with controlled conditions and with free access to food and water. The male reproducers were on
commercial diet until mating, while the females were either on a control diet (CON) or on an EFAD since 3 weeks before mating
until end of lactation. CON (soy oil) had 2.7% of linoleic acid (LA) and 0.05% of &ALPHA;-linolenic acid (LNA) while EFAD
(coconut oil) had 0.01% of LA, being practically free of LNA. The offspring were reduced to 8 males/mother on Day 3 of life
being weaned on commercial diet until adults. They were separated according to the maternal diet in CON (n=11) and EFAD
group (n=15). As adults, the offspring were housed for 24h in metabolic cages (MC) with free access to food and water, in order
to calculate: the food (FI) and water (WI) ingestions; the urine volume (V), density (D), Na + , K+, protein, and creatinine
excretions by 100g/body weight. Indirect Systolic Blood Pressure (SBP) was taken 5 times per day in the adult offspring.
Glomerular filtration and proximal tubule Na + reabsorption were measured respectively by creatinine and Li + clearances for 3h in
the MC, in the adult offspring previously water expanded at 5 mL/100g. Blood and urine samples were collected, in order to
calculate the excretions and clearances. Heart, lungs, liver, spleen, kidneys and testicles were removed and weighed (wet weight),
dried for 48h at 70oC and then, they were subsequently weighed (dry weight) with values being corrected to 100g/body weight.
Results are in meanSEM. Offspring from the EFAD group weighed less than CON respectively at: birth (6.10.1 vs 5.60.1* g;
CON vs EFAD), weaning (520.8 vs 461.1* g; CON vs EFAD), growth (203.51.5 vs 193.72.5* g; CON vs EFAD) and adult
(3205.0 vs 298.78.0* g; CON vs EFAD). At the 24h adult offspring studies: FI (6.80.2 vs 7.10.2 g/100g; CON vs EFAD);
WI (11.50.5 vs 10.90.3 mL/100g; CON vs EFAD); V (4.80.3 vs 4.60.3 mL/100g; CON vs EFAD), a D (1.0460.001 vs
1.0490.0005 g/mL; CON vs EFAD), K+ (5499463.6 vs 5242512.9 Eq/mL/100g; CON vs EFAD) and creatinine urinary
excretions (4.10.3 vs 4.10.2 mg/mL/24h; CON vs EFAD) were similar between groups but Na+ (416.521.5 vs 348.321.6*
Eq/mL/100g; CON vs EFAD) and protein excretions (17.91.3 vs 12.30.8* mg/100g/24h; CON, n= 10 vs CAGE, n= 8) were
decreased in the EFAD. PAS measured in adult offspring (1490.64 vs 1470.9 mmHg; CON, n= 12 vs EFAD, n= 9) was similar
between groups. The lack of difference on creatinine clearance (220.512.6 vs 212.815.7 L/min/100g; CON, n= 9 vs EFAD,
n= 8) stands for an unaltered glomerular filtration by the diet. The decrement in Li+ clearance (62.03.2 vs 37.37.9*
L/min/100g; CON, n= 8 vs EFAD, n=7) indicates increased Na + proximal tubule reabsorption. Organ wet and dry weights were
similar among CON and EFAD adult offspring.
Conclusions:
Maternal amplified perinatal EFAD reduced the weight gain from birth and altered renal functional aspects in the adult offspring.
Keywords: ESSENTIAL FATTY ACID DEFICIENCY , PROXIMAL TUBULE REABSORPTION, RENAL FUNCTION
Financial Support: PROCAD 8052 and FACEPE/IBPG-0143-2.07/09
Resumo:07-018
EARLY GLOMERULAR CHANGES INDUCED BY HIGH-FAT DIET IN OVARIECTOMIZED RATS.
Amaral, L. S. B. 1; Silva, F. A. 1; Ribas, W. B. D. 1; Silva, J. A. 1; Souza, S. I. 1; Magalhes, A. C. M. 1;

Macedo, C. L. 2; Coimbra, T. M. 3; Soares, T. J. 1
1
Instituto Multidisciplinar em Sade/UFBA , IMS/UFBA
2
Departamento de Cincias Naturais/UESB, DCN/UESB
3
Departamento de Fisiologia, FMRP/USP
Objectives:
The association between obesity and estrogen deficiency may contribute to the changes of renal function and structure. Previous
studies in our laboratory have demonstrated increased blood pressure and visceral fat depot, decreased fractional excretion of
sodium and tubulointerstitial lesions in ovariectomized rats fed a high-fat diet. The aim of this study was to evaluate the effects of
high-fat diet on urinary protein and albumin excretion and glomerular changes in ovariectomized rats.
Twenty-four ten-week-old female Wistar rats were ovariectomized (OV) or sham-operated and divided into four groups: SSD
(sham operated rats with standard diet, n=6); OSD (OV rats with standard diet, n=6); SHFD (sham operated rats with high-fat
diet, n=6); OHFD (OV rats with high-fat diet 54.4% of total calories from fat, n=6). Monthly urine samples were collected to
quantify albumin and protein excretion. The quantification of urinary protein excretion was performed by colorimetry. Urinary
albumin excretion was quantified in ovariectomized and sham animals by electroimmunoassay, using a specific antibody against
rat albumin. The animals were killed 24 weeks after the diet, at which time the kidneys were removed for histological and
morphometric analysis. The slices of renal tissue were stained with Masson's trichrome and examined under light microscopy.
Glomerulosclerosis was evaluated semiquantitatively in 100 glomeruli per animal by counting the number of quadrants showing
sclerosis. The tissue was also used for morphometric studies of glomerular size. Measurements of the glomerular tuft area were
done on 30 consecutive cortical glomeruli and 15 corticomedullary glomeruli of each animal from different experimental groups.
Data concerning albuminuria and proteinuria were submitted to the KruskalWallis nonparametric test. Other data were
submitted to the Newman-Keuls test. The level of significance considered was p
Conclusions:
Our results suggest that high-fat diet and ovariectomy may be contributing to the proteinuria and glomerular sclerosis and
hypertrophy in these animals.
Keywords: Glomeruloesclerose, Proteinria, Dieta hiperlipdica, Ovariectomia
Financial Support: CNPQ and FAPESB
Resumo:07-019
INFLUENCE OF HAEMODIALYSIS AND HEPATITIS C ON BLOOD OXIDATIVE STATUS IN PATIENTS OF
DIALYSIS UNITS IN RIO GRANDE/RS-BRAZIL.
Kalb, A. C. 1; Silva, N. M. O. D. 2; Martnez, P. E. 1; Martnez, A. M. B. 2

1
Instituto de Cincias Biolgicas, FURG
2
Faculdade de Medicina, FURG
Objectives:
The uremic state and the bio-incompatibility of haemodialysis (HD) are associated with oxidative stress in HD patients. High
prevalence of hepatitis C virus (HCV) has been found in many dialysis units around the world, giving to HD patients an increased
probability to acquire this infection. This study aimed verifies the influence of haemodialysis and HCV infection on blood
oxidative status in patients of dialysis units in Rio GrandeRSBrazil.

It was measured oxidative stress markers in blood cells and plasma, and blood collection were performed before haemodialysis
procedure. The following groups were considered: 1.Control subjects (CTRL); 2.Haemodialysismaintened patients (HD);
3.HCVinfected patients (HCV) and 4.Haemodialysismaintened patients infected by HCV (HCVD) (n=14 per group). In
erithrocytes, was verified nonprotein sulfhydryl (SH) content (which include the antioxidant GSH), and lipid peroxidation. In
leucocytes, it was measured total antioxidant capacity against peroxyl radicals. It was also verified lipid damage in plasma
samples. Statistical analysis was performed through nonparametric KruskalWallis test where data was expressed in medians
and interquartile range (2575). Results: 1. In erithrocytes, SH content of all groups of patients presented absence (p>0.05) of
statistical differences when compared to CTRL group (CTRL: 0.00067 (0.0005210.000809); HD: 0.000691
(0.0005300.001016); HCV: 0.000462 (0.0002920.000508); HCVD: 0.000587 (0.0003340.000890)); however, there was a
significant difference (p
Conclusions:
Probably due to successive stimuli caused by dialysis, HD patients had slightly higher basal levels of GSH in erithrocytes, which
although subtle, were sufficient to decrease the observed values of lipid peroxidation. These stimuli could also provide an
improvement in antioxidant competence in leucocytes. However, it seems that HCV in some way blocks this compensatory
response, once HCVD patients presented high content of lipid damage both in red cells and plasma. The results suggest that
HCVinfected patients submitted to dialysis in Rio Grande are quite more susceptible to oxidative damage than only
HCVinfected or only haemodialysismaintened patients, and this fact implicates a more careful clinical approach.
Keywords: Haemodialysis, Hepatitis C, Oxidative status
Resumo:07-020
QUANTITATIVE EVALUATION OF LEUKOCYTES AND PHYSICAL PARAMETERS IN RATS WITH
NEPHROTIC SYNDROME INDUCED BY DOXORUBICIN HYDROCHLORIDE
Almeida. 1; Melo1; Jardim. 1; Domingues1; Moreira 1; Santos1; Melo1; Silva2; Pereira1

1
Universidade Federal dos Vales do Jequitinhonha e Mucuri, UFVJM
2
Universidade Federal de Minas Gerais, UFMG
Objectives:
The present study intended achieve a quantitative analysis of rat peripheral blood leukocytes with NS induced by doxorubicin as
well as evaluate some physical parameters.
It was used 40 male Wistar rats, average weight of 300 grams. The animals were housed in individual cages, and have received a
standard commercial diet and water ad libitum. We share the animals into two groups: DOXO group (n=20), underwent
intravenous injection of doxorubicin (Doxolem/7.5mg/kg body weight) and SAL group (n=20), which received intravenous
injection of saline (D. Boonsanit et al, 2006). To confirmation of renal disease, microalbuminuria tests were performed - cobas
mira plus device with use of kits BIOCLIN /Quibasa . To comparison between groups test t student was performed.
Significant level of *p < 0,05 was established. The animals in each group were analyzed 7, 14, 21 and 28 days after the injections.
Urine was collected for 24 hours (metabolic cage) on day before sacrifice. On the sacrifice day, animals were weighed and
underwent thoracotomy under anesthesia (ketamine / Xylazin - 70/10 mg / kg). Blood samples were collected by cardiac
puncture. After specific lyses of the red cells, the absolute leukocytes count was performed using Newbaeur Chamber. The
organs were perfused with buffered saline (PBS) and the kidneys were removed and immediately weighed. The weight of the
kidneys was normalized using body weight. According to our results, the DOXO group presented a decrease of the absolute
number of leukocytes 7 days (2309 322,5 vs. 3740 423.4 cells number, p < 0,001) after the injection however on 14th after
injection the same group presented an increased of this parameter (6475 249,8 vs. 4270 250.1mL, p < 0,0001). In the first
three weeks the animals from DOXO presented a reduction in water consumption, but no statistical differences was observed.
Similar results were observed in relation to urine volume. Regarding body weight, it was observed that DOXO group kept or
reduced weight, differing from the controls which gained weight over weeks, with significant statistical difference between
groups for days 14 (277.9 11.61 vs. 343.2 13.98g, p < 0,007), 21 (296.1 11.49 vs. 389.4 15.27g, p < 0,0005) and 28
(308.2 14.69 vs. 379.0 8.895g, p < 0,004). About kidney weight in the DOXO group there was an increased value in weight
after the second week of injection, 14 (5.354 0.16 vs. 4.388 0.14g, p < 0,005), 21 (6.408 0.17 vs. 4.309 0.10g, p <
0,0001), 28 days (6.921 0.29 vs. 4.413 0.18g, p < 0,0001).
Conclusions:
We believe that increasing the number of leukocytes observed in the 14 th day of injury is possibly related to the inflammatory
response established in renal tissue. We confirmed some changes in the body, as reported in the literature, indicating the
progressive nature of the disease. Other immunophenotyping experiments are being conducted to confirm or clarify the behavior
of the immune system in different stages of the disease.
Keywords: Doxorubicin, Nephrotic Syndrome, Proteinuria
Resumo:08-043
DISTRIBUTION OF O2 CHEMORECEPTORS AND CONTROL OF CARDIORESPIRATORY RESPONSES TO
HYPOXIA IN NILE TILAPIA (OREOCHROMIS NILOTICUS)
Zeraik, V. M. ; Belo, T. C. ; Sanches, J. R. ; Kalinin, A. L. ; Rantin, F. T.

Departamento de Cincias Fisiolgicas, UFSCar
Objectives:
In fish the O2 chemoreceptors are mainly found in the gill arches and monitor the O2 tensions of the inspired water (externally
oriented) and/or the arterial blood (internally oriented). During hypoxia these receptors trigger cardiorespiratory adjustments in
order to maintain an adequate O2 transfer to the tissues. The aim of the present study was to determine the O2 chemoreceptors
distribution through the gill arches of Nile tilapia and the role of these receptors on the cardiorespiratory responses to hypoxia in
this species.
One buccal and two opercular canullae, and a pair of ECG electrodes were implanted in a group of intact fish (control group, n =
10; Wt = 258,46 7,42 g). This procedure served to measure the following cardiorespiratory variables: respiratory frequency fR, ventilatory tidal volume VT, gill ventilation - VG, O2 extraction from the ventilatory current EO2, oxygen uptake - VO2,
and heart rate fH. After an overnight recovery period from surgery fish were subjected to the following O2 tensions during 40
min each: 140 (normoxia), 100, 70, 50, 30 e 20 mmHg. To verify the O2 receptors distribution in the gills, the first pair of gill
arches was surgically extirpated (operated group, n=10; Wt = 232,53 8,78 g). After a recovery period of 4 days, operated fish
were subjected to the same procedures described above for the intact ones. VO2 of intact fish was significantly higher than that of
operated in all experimental O2 tensions. In normoxia the fR of the operated fish was 30% higher than that recorded for the
control group. During graded hypoxia this variable was even higher (44% at 50 mmHg) in the operated group. No significant
differences were observed in VT and VG of both groups, except in the most hypoxic tension of 20 mmHg, when the operated
group presented a significant lower VG. The ventilarory requirement (VG/VO2) was significantly higher in operated fish during
normoxia, at 100 mmHg and 20 mmHg (more hypoxic tension). The fH of operated fish was significantly higher in normoxia and
at 100 mmHg. Below these tensions both groups presented similar fH. In both groups EO2 was maintained within a range of 70 to
80%. Intact fish presented significantly higher EO2 only during normoxia.
Conclusions:
In Nile tilapia the O2 chemoreceptors are distributed beyond the first gill arch, probably through the four gill arches. These
receptors trigger the cardiorespiratory adjustments necessary to face the hypoxic conditions at which the species is frequently
subjected in its natural environmental conditions.
Keywords: brnquias, hipxia, quimiorreceptores, respostas cardiorrespiratrias, Tilpia-do-Nilo
Resumo:08-044
OXIDATIVE DAMAGE INDUCED BY CIGARETTE SMOKE EXPOSURE AFFECTS SKELETAL MUSCLE
BEFORE LUNG TISSUE
Alves, E. C. 1; Nesi, R. T. 6; Carlos, S. P. 1; Silva, L. A. 1; Effting, P. S. 1; Valena, S. V. 6; Pinho, R. A. 1;

Tuon, L. 1; Chiappa, G. R. 1,10
1
Universidade do Extremo Sul Catarinense, UNESC
6
10
Hospital das Clinicas de Porto Alegre, HCPA
Objectives:
Studies have shown that the cigarette smoke (CS) is responsible for pathogenesis of several pulmonary diseases including chronic
obstructive pulmonary disease (COPD). It is well known that cigarette smoke is responsible for lung damage, inflammation and
oxidative stress in pulmonary emphysema. In addition, COPD is frequently associated with muscle loss and skeletal muscle
dysfunction. For this reason, the present study investigated the inflammation and oxidative stress in mice exposed to cigarette
smoke for 7, 15, 30, 45 and 60 consecutive days to know when the oxidative damage in skeletal muscle happened, with acute or
chronic CS exposure.
Thirty-six C57BL-6 mice were divided into six groups (n = 6): control group, 7-, 15-, 30-, 45- and 60-day exposure to CS,
respectively. The CS groups were exposed to cigarette smoke 3 times/day (4 cigarettes/session). Twenty-four hours after the last
cigarette exposure, the animals were sacrificed and used for subsequent assays. In addition to the analysis of lipid and protein
oxidation (TBARS and Carbonylation), sulfhydryl assay, histological and morphological studies were performed. The data
demonstrated the relationship between the oxidative damage measured through lung, diaphragm and quadriceps tissues.
Conclusions:
Our data demonstrated the influence of CS in oxidative damage not only in lung, but in skeletal muscle, showing the diffusion
effect of CS and that skeletal muscle are affected before the lung tissue.
Keywords: cigarette smoke, mice, Oxidative damage, skeletal muscle
Financial Support: Universidade do Extremo Sul Catarinense- UNESC
Resumo:08-045
ESTRADIOL MEDIATES THE ACUTE LUNG INFLAMMATION AFTER INTESTINAL ISCHEMIAREPERFUSION (I/R) IN FEMALE AND MALE RATS.
Fantozzi, E. T.
Farmacologia/ICB, USP
Objectives:
Sexual dimorphism interferes with Th1 and Th2 lymphocyte functions, and accordingly estradiol (e2) plays a role on developing
acute lung injury (ali) caused by intestinal I/R. Female rats are more resistent to organ injury caused by hemorrhagic shock. Nitric
oxide (NO) exerts pro and antinflammatory effects in gut trauma-induced lung injury, but its interaction with estradiol is still not
clear. Here we assessed the effect of E2 on the lung and intestinal inflammation after intestinal I/R and investigated the putative
involvement of inducible NOS (iNOS) pathway.
Anesthetized wistar male and female rats (60 days, 200g) were subjected to occlusion of the superior mesenteric artery (SMA) for
45 min, followed by 2 hr of reperfusion. Groups of rats (n=5) were treated with E2 (17 estradiol, 280g/kg, s.c. 24 h before or
30 min (i.v.) after induction of ischemia. In parallel, rats were treated with selective iNOS inhibitor, aminoguanidine (50 mg/kg
iv) or with L-arginine (300mg/kg, i.p.) 1h before arterial occlusion. Lung and intestinal vascular permeability (LVP and IVP)
were assessed by Evans blue dye extravasation and neutrophil recruitment to the tissues by the myeloperoxidase (MPO) method.
Lung generation of IL-10 was also assessed by ELISA in cultured lung tissue. 7-days-ovariectomized (OVx) rats had LVP, IVP
and MPO increased after I/R whereas IL-10 levels were significantly reduced. E2 treatment reverted the increased LVP, but not
IVP nor MPO. Aminoguanidine alone reduced LVP in OVX rats, but failed to do so when associated to E2. Aminoguanidine in
non-OVx rats markedly exacerbated LVP and IVP. Male rats treated with E2 were protected with a significant reduction of LVP.
Conclusions:
Endogenous NO from iNOS, likely reduces the vascular effects of intestinal I/R. In addition, L-arginine imitated E2 since it
reduced ALI after intestinal I/R. In conclusion, estradiol reduces lung inflammation after intestinal I/R, but the concomitant
blockade of NOS suppresses this protection.
Keywords: Intestinal isquemia-reperfusion , Pulmonary inflammation, Rats, Estradiol, Nitric oxide
Financial Support: CNPq and FAPESP.
Resumo:08-046
INFLUENCE OF DIFFERENT PHASES OF ESTROUS CYCLE IN THE VENTILATORY RESPONSES TO
HYPERCAPNIA AND HYPOXIA IN RATS
Marques, D. A. 1,1; Gargaglioni, L. H. 1; de Carvalho, D. . 1; Szawka, R. E. 2; Franci, J. A. 3; Bcego, K. C. 1

1
Morfologia e Fisiologia Animal, UNESP, FCAVJ
3
Laboratrio de Neuroendocrinologia I, USP
2
Departamento de Fisiologia e Biofsica, UFMG
Objectives:
Determine the respiratory and thermoregulatory responses to hypoxia and hypercapnia in rats at each stage of the estrous cycle
The present study examined the hypoxic (HVR) and hypercapnic (HCVR) respiratory responses of female rats weighing 250300g, in different phases of estrous cycle: estrus, diestrus, proestrus and also in ovariectomized rats. Ventilation was measured
using a whole-body plethysmograph (Bartlett and Tenney, 1970). Vaginal smears were taken daily to verify estrous cycle
regularity and only rats showing at least five consecutive regular 4-day estrous cycles were included in this study. Pulmonary
ventilation (VE) and body temperature (Tb) were measured followed by 30 min of hypercapnic exposure (7% CO2 in air) and
hypoxia (5% O2) in unanesthetized animals. Under resting conditions, there is no difference in VE between animals. Tb was
greater in estrus phase compared to other groups during air and 7%CO2. Hypoxia caused an increase in VE and a drop in Tb in
all groups, but the HVR was smaller in diestrus (717.8 41.5 mL/Kg/min), proestrus (712.1 34.1 mL/Kg/min), and in
ovariectomized (516.6 13.4 mL/Kg/min) rats compared to estrous phase (1213.2 284 mL/Kg/min), due to a reduced tidal
volume. Hypoxia-induced drop in Tb was not different between groups. HCVR was also reduced in diestrus (978.6 88.9
mL/Kg/min), proestrus (1056.7 108.9 mL/Kg/min), and in ovariectomized rats (717.3 71.2 mL/Kg/min), due to a decreased
tidal volume. Thus, the different phases of the estrous cycle influence the respiratory responses to hypercapnia and hypoxia in
rats, acting on tidal volume. Additionally, Tb is increased in estrous cycle during normoxia, normocapnia and hypercapnia but is
not different in hypoxic challenge.
Conclusions:
The different phases of the estrous cycle influence the respiratory responses to hypercapnia and hypoxia in rats, acting on tidal
volume.
Keywords: estrous cycle, ventilatory responses, hypercapnia, hypoxia
Resumo:08-047
EFFECTS OF OLEANOLIC ACID ON PULMONARY MORPHOFUNCTIONAL AND BIOCHEMICAL VARIABLES
IN EXPERIMENTAL SEPSIS
Ramos, M. B. D. A. 1; Santos, R. S. 1; Silva, P. L. 1; Oliveira, G. P. D. 1; Cruz, F. F. 1; Assis, E. F. D. 2;

Castro-faria-neto, H. C. D. 2; Gattass, C. R. 1; Rocco. , P. R. M. 1
1
Instituto de Biofsica Carlos Chagas Filho, IBCCF
Instituto Oswaldo Cruz , IOC
Objectives:
Sepsis and its associated complications, such as acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) and multiple
organ failure, present high morbidity and mortality despite recent advances in therapeutic strategies. Recently, natural products
derived from plant extracts and their synthetic derivatives are being increasingly used to treat a wide range of diseases due to
their anti-inflammatory property. In this line, oleanolic acid (OA), a triterpenoid compound present in a great variety of plants
and food products, modulates the production and activity of pro-inflammatory cytokines and enzymatic antioxidant defense. The
aim of the present study was to test the hypothesis that OA may curtail the inflammatory process, improving lung morphology
and function in experimental sepsis.
36 BALB/c mice were randomly assigned into two groups: a) Sepsis was induced by cecal ligation and puncture (CLP) surgery,
and b) Sham operated group was used as control. One-hour after surgery, Sham and CLP groups were further randomized into
subgroups receiving saline (SAL), OA (10 mg/kg ip), or dexamethasone (DEXA, 1 mg/kg ip). After 24 hours, animals were
anesthetized, and static lung elastance (Est, L), lung histology (light microscopy), levels of interleukin (IL)-6, and IL-8 in the
bronchoalveolar lavage fluid and the degree of cell apoptosis in lung and kidney were analyzed. Est, L (55%) and alveolar
collapse (75%) were higher in CLP-SAL group compared to Sham-SAL (p< 0.05). Furthermore, CLP-SAL group compared
showed interstitial edema, polymorphonuclear cell infiltration, and high degree of epithelial cell apoptosis in lung and kidney
compared to Sham-SAL. The treatment with OA and DEXA reduced Est, L (35%, and 42%, respectively) and alveolar collapse
(70%, and 64%) in relation to CLP-SAL group. The OA and DEXA treatment minimized the polymorphonuclear cell infiltration
and the degree of apoptosis in lung and kidney compared to CLP-SAL group. However, DEXA, but not OA reduced the levels of
IL-6, and IL-8 (p<0.05).
Conclusions:
In the present model of sepsis, both OA and DEXA improved lung morpho-function, as well as acted on distal organs.
Nevertheless, the mechanisms related to these beneficial effects differ in OA and DEXA. Therefore further studies are required to
better elucidate the mechanism of action of OA in sepsis.
Keywords: Acute Lung Injury, Interleucin, Histology, Lung mechanics, Oleanolic Acid
Financial Support: CNPq, PRONEX-FAPERJ, FAPERJ, CAPES, INCT-INOFAR
Resumo:08-048
FREQUENCY OF TIME-CYCLED CONTROL BREATHS DURING BIPHASIC POSITIVE AIRWAY PRESSURE
MODIFIES THE EXPRESSION OF LUNG INFLAMMATORY AND FIBROGENIC MEDIATORS IN PULMONARY
AND EXTRAPULMONARY ACUTE LUNG INJURY.
Saddy, F. ; Moraes, L. ; Oliveira, G. P. ; Santos, C. L. ; Cruz, F. C. ; Morales, M. M. ; Garcia, C. S. ; Pelosi,

P. ; Rocco, P. R.
Laboratrio de Investigao Pulmonar/IBCCF, UFRJ
Objectives:
To compare the effects of different frequencies of time-cycled control breaths during biphasic positive airway pressure (BiVent)
with pressure controlled ventilation (PCV) on lung histology, arterial blood gases, inflammatory and fibrogenic mediators in
models of pulmonary (p) and extrapulmonary (exp) acute lung injury (ALI) with similar transpulmonary pressures.
Wistar rats were randomly divided into two groups. In ALI groups, animals received Escherichia coli lipopolysaccharide
intratracheally (100 microg, ALIp) or intraperitoneally (1 mg, ALIexp). After 24 h, animals were anesthetized and further
randomized as follows: 1) pressure controlled ventilation mode (PCV) with tidal volume (VT)=6 ml/kg and inspiratory to
expiratory ratio (I:E)=1:2; 2) BiVent with three different time cycles (controlled/spontaneous breath cycles 50, 75 and 100).
BiVent was set with two levels of CPAP [inspiratory pressure (PHigh=10cmH2O) and positive end-expiratory pressure
(PLow=5cmH2O)] and the inspiratory time was sustained as THigh=0.3s. All rats were mechanically ventilated with FiO2=0.4
and PEEP=5cmH2O for 1 hour. BiVent yielded better functional improvement and less lung injury compared to PCV
independent of ALI etiology. BiVent 50 improved gas exchange and reduced atelectasis compared to PCV, BiVent75 and
BiVent100 in both ALI groups. Interleukin (IL)-6, IL-1 beta, and type III procollagen (PCIII) expressions were lower in BiVent
compared to PCV. BiVent 50 presented lower mRNA expression of IL-6, IL-1 beta, and PCIII compared to PCV, BiVent75, and
BiVent100 in both ALI groups.
Conclusions:
BiVent presented greater beneficial effects on respiratory function and a reduction in lung injury compared to PCV. Among the
different frequencies of BiVent, BiVent50 demonstrated better functional results with less lung damage and expression of
inflammatory mediators independent of ALI etiology.
Keywords: Mechanical ventilation, Ventilator induced lung injury, Acute lung injury, Inflamatory mediators
Financial Support: PRONEX, FAPERJ, CNPq
Resumo:08-049
PULMONARY AND LIVER INJURY AFTER CHRONIC EXPOSURE TO SUBLETHAL DOSES OF MICROCYSTINLR.
Carvalho, G. M. C. 1; Oliveira, V. R. D. 1; Casquilho, N. V. 1; Soares, R. M. 1; Azevedo, S. 1; Pires, K. M. 2;

Valena, S. S. 2; Zin, W. A. 1
1
2
Universidade do Estado do Rio de Janeiro, UERJ
Objectives:
Biological pollution caused by cyanotoxins leads to respiratory function impairment. Thus, we aimed to study pulmonary
mechanics, lung and liver histology in mice chronically submitted to sublethal doses of microcystin-LR (MCLR) and evaluate
whether the results depended on the doses.
Male Swiss mice received ten doses of distillated water intraperitoneally (ip, 100 mL, CTRL n=6) or sublethal doses of MCLR
(5, 10, 15 and 20 g/kg ip in 100 mL of distilled water, TOX n=24) on alternate days. 24 h after the last injection pulmonary
mechanics [static elastance (Est), viscoelastic component of elastance (E), resistive (P1), viscoelastic (P2), and total (Ptot)
pressures] were determined, and lungs and livers were prepared for histopathology. ANOVA was used to test differences among
the groups. Est, E, P1, P2 and Ptot were significantly higher than CTRL [25.7 0.60 cmH2O/mL; 3.73 0.14
cmH2O/mL; 0.61 0.04 cmH2O; 0.75 0.03 cmH2O; 1.36 0.07 cmH2O (mean SEM)] in TOX10 (30.9 1.40 cmH2O/mL;
5.1 0.41 cmH2O/mL; 0.9 0,12 cmH2O; 1.1 0.09 cmH2O; 2.0 0.17 cmH2O), TOX15 (33.6 1.59 cmH2O/mL; 4.9 0.36
cmH2O/mL; 0.8 0.02 cmH2O; 1.0 0.06 cmH2O; 1.8 0,06 cmH2O) and TOX20 (38.9 4.8 cmH2O/mL; 5.4 0.74
cmH2O/mL; 0.9 0.07 cmH2O; 1.1 0.13 cmH2O; 2.0 0.21 cmH2O), but did not differ among them. TOX5 (30.4 2.25
cmH2O/mL; 4.3 0.23 cmH2O/mL; 0.7 0.01 cmH2O; 0.9 0.03 cmH2O; 1.5 0.04 cmH2O) was similar to CTRL in all
parameters. Alveolar collapse was higher in TOX15 (22.2%) and TOX20 (32.6%) than in CTRL (10.8%). The lung inflammatory
cell content (cells/m2) increased in all MCLR doses, but did not differ among them: TOX5=1.88 x 10-3, TOX10=2.07 x 10-3,
TOX15=2.19 x 10-3 and TOX20= 2.17 x 10-3 in relation to CTRL= 0.79 x 10-3. The liver weight was significantly higher than
CTRL= 1.75 g in TOX5= 2.06 g, TOX10= 2.12 g, TOX15= 2.19 g and TOX20= 2.15 g, but did not differ among them. All TOX
mice showed a complete loss of liver architecture with hepatomegaly, steatosis, dilated sinusoidal spaces, necrosis, inflammatory
foci and a high degree of binucleated hepatocytes.
Conclusions:
Chronic exposure to MCLR impaired pulmonary mechanics, lung and liver histology. These findings did not depend on the
degree of exposure.
Keywords: Chronic Exposition, Liver Histology, Lung Histology, Microcystin-LR, Respiratory Mechanics
Financial Support: FAPERJ, CNPq, MCT.
Resumo:08-050
ANTIOXIDANT TREATMENT PROTECTS AGAINST MATRIX METALLOPROTEINASES ACTIVATION AND
CARDIOMYOCYTE INJURY DURING ACUTE PULMONARY THROMBOEMBOLISM.
Santos-o. S1; Neves-e. M. N. 1; Ferraz, K. C. 2; Ceron, C. S. 1; Rizzi, E. 1; Gerlach, R. F. 3; Santos- J. E. T. 1

3
Department of Morphology, Estomatology and Physiology,, FORP-USP
2
Department of Pharmacology, State University of Campinas, UNICAMP
1
Department of Pharmacology, Faculty of Medicine of Ribeirao , FMRP-USP
Objectives:
Increased reactive oxygen species (ROS) promote matrix metalloproteinases (MMPs) activities and may underlie cardiomyocyte
injury and degradation of cardiac troponin I (cTI) during acute pulmonary thromboembolism (APT). We examined whether
pretreatment with tempol (a general ROS scavenger) prevents MMPs activation and protects against cardiomyocyte injury of
APT. We have also examined the therapeutic effects of tempol administration after APT.
Hemodynamic evaluations were performed in non-embolized sheep treated with saline (n=4) and in sheep that were embolized
with autologous blood clots (n=8) and that received continuous infusion of tempol (1.0 mg/kg/min) before (n=7) and after
induction of APT (n=7). The activity of MMPs and reactive oxygen species in right ventricle, as well as the levels of serum TnIc
were measured by zymography, DHE and enzyme immunoassay, respectively. APT increased the mean pulmonary arterial
pressure and pulmonary vascular resistance by approximately 146% and 164%, respectively (both P
Conclusions:
We found evidence supporting the idea that RV oxidative stress after APT increases MMPs activities in the RV leading to
degradation of sarcomeric proteins including cTI. Antioxidant treatment may prevent MMPs activation and protect against
cardiomyocyte injury after APT.
Keywords: acute pulmonary thromboembolism, matrix metalloproteinases, oxidative stress, pulmonary hypertension, troponin
Resumo:08-051
PULMONARY AND RENAL EFFECTS OF INTRAVASCULAR VOLUME REPLACEMENT IN A NON-SEPTIC
MODEL OF ACUTE LUNG INJURY
Silva, P. L. 2,1; Gldner, A. 1; Uhlig, C. 1; Carvalho, N. R. 1; Beda, A. 1; Rentzsch, I. 1; Kasper, M. 3; Pelosi,

P. 4; Rocco, P. R. M. 2; Abreu, M. G. D. 1
2
Laboratory of Pulmonary Investigation, IBCCF, UFRJ
1
1Department of Anesthesiology and Intensive Care Therapy, TU Dresden
3
Institute of Anatomy, TU Dresden
4
Dipartimento di scienze chirurgiche e diagnostiche integrat, University of Genua
Objectives:
Intravascular volume replacement may be necessary during acute lung injury (ALI). The aim of the present study was to compare
the effects of Ringers acetate (RA), gelafundin (GEL), and hydroxyethyl starch (HES) on lung and renal damage in experimental
ALI.
Non-septic ALI was induced in pigs (363 kg) by lung saline lung lavage followed by ventilation with high tidal volumes.
Protective ventilation was then initiated and 25% of the estimated blood volume was drawn. The intravascular volume was
partially replaced with RA, GEL or HES (n=10/group) to achieve 90% of the intrathoracic blood volume index (ITBVi) before
blood drainage. The amount of fluid to achieve the target ITBVi was higher with RA (2250764 ml) than GEL (704159 ml) and
HES (83782 ml). Gas exchange and hemodynamic variables did not differ among groups. Lung elastance was higher in RA and
GEL than HES. The W/D ratio was more elevated in RA (7.91.0) than GEL (6.50.5) and HES (6.50.7), (p
Conclusions:
In this non-septic ALI model, volume replacement with RA worsened lung mechanics and histology as well as pro-inflammatory
lung response and edema compared to GEL and HES. However, GEL led to higher acute kidney injury score than other groups.
Keywords: Acute Lung Injury, Intravascular Volume, Lung Histology, Mechanical Ventilation, Wet-to-Dry Ratio
Financial Support: DFG, CAPES-DAAD, CNPq, PRONEX, FAPERJ
Resumo:08-052
EFFECTS OF DIFFERENT TYPES OF POLYMERIC LIPOSOMES USED FOR GENE DELIVERY ON LUNG
MECHANICS AND HISTOLOGY
Xisto, D. G. 1,2; Abreu, S. C. 1; Silva, J. D. 1; Silva, A. L. 1; Crossetti, J. 1; Martini, S. V. 2; Temprana, C. F.

3
; Alonso, S. V. 3; Rocco, P. R. M. 1; Morales, M. M. 2
1
2
Laboratory of Cellular and Molecular Physiology, IBCCF, UFRJ
3
LBM, Department of Science and Technology, UNQ
Objectives:
The benefits of gene therapy for the treatment of respiratory diseases have been disappointing due to viral vectors toxicity and
immunogenicity. Consequently, the need for safer alternatives has led to the development of polymeric liposomes. However,
some particles may also lead to toxicity limiting their use. The aim of this study was to analyze the impact of different polymeric
liposomes on lung mechanics and histology.
48 BALB/c mice were randomly divided into 6 groups, which were intratracheally instilled with 50 ul of 5 mM of the following
suspensions: DMPC:DC8,9PC:DOTAP, polymerized (A), DMPC:DC8,9PC:SA, polymerized (B), DMPC:DC8,9PC: miristoyl
choline chloride (MCl), polymerized (C), DMPC:DC8,9PC:DOTAP, non-polymerized (D), DMPC:DC8,9PC:SA, nonpolymerized (E) and DMPC:DC8,9PC: MCl, non-polymerized (F). At twenty-four hours, the animals were anesthetized,
tracheotomized and mechanically ventilated. Lung static elastance, resistive and viscoelastic/inhomogeneous pressures were
analyzed by end-inflation occlusion method. Lungs were fixed and stained for histopathological analysis. Resistive and
viscoelastic pressures and static elastance were similar in all groups, as well as tissue cellularity. However, animals treated with
non-polymerized particles presented higher mortality rate (60%).
Conclusions:
The present polymeric liposomes particles did not impair lung mechanics, histology or increase tissue cellularity. However, nonpolymerized liposomes particle presented higher systemic toxicity with greater mortality.
Keywords: liposomes, lung mechanics, lung histology
Financial Support: FAPERJ, CAPES, CNPq, PRONEX-FAPERJ
Resumo:08-053
DIAPHRAGMATIC MOTION STUDIED BY B-MODE ULTRASONOGRAPHY IN MORBID OBESE SUBJECTS
Tenrio, L. H. S. 1; Amaral, F. J. 2; Neto, J. B. C. 3; Passos, V. M. M. 1; Santos, A. C. 5; Lima, A. M. J. 4;

Brasileiro-santos, M. S. 5
Fisioterapia, UFPE
Radiologia/Hospital Baro de Lucena, HBL
3
Cirurgia geral/Hospital Agamenon Magalhes, HAM
4
Departamento de Morfologia e Fisiologia Animal, UFRPE
5
Educao Fsica, UFPB
2
Objectives:
Diaphragm mobility can be assessed by several images techniques, however there is no data about the assessment of diaphragm
mobility by ultrasound in morbid obese patients. Therefore, the aim of this study was to evaluate diaphragm mobility by
ultrasound B-mode in individuals with morbid obesity.
Thirty-one patients were evaluated of both genders, with mean age of 35.5 9.7 years, body mass index of 43.83.3 Kg/m2 and
with no previous history of cardiac or respiratory illness. The diaphragm mobility (DM) was assessed by the ultrasound Philips
HD7 (Philips Medical Systems, Bothell, WA, USA) with a convex transducer of 2.-5MHz of high resolution in B-mode. The DM
was observed in three moments: quiet breath (QB), voluntary sniffing (VS) and deep breathing (DB) as we used the liver window
on the right side to assess these three moments. Data distribution was analyzed by the Kolmogorov-Smirnov test and analysis of
variance (ANOVA) of one factor. Results are expressed as means standard deviation and as 95% confidence limits. The DM
was assessed with success during the QB, VS and DB on the right hemidiaphragmatic side in all obese subjects. After inferential
analysis it was observed significant differences at DM during the QB and VS (1.870.49cm vs. 3.491.17 cm, p<0,001).
Conclusions:
According to the results of this work was shown that B-mode ultrasonography is a reproducible method to assess the hemi
diaphragmatic motion in the morbidly obese.
Keywords: Ultrasonography , Morbid Obesity, Diaphragm
Resumo:08-054
IMPACT OF EXERCISE ON INFLAMATORY PROCESS AND REMODELING IN A MURINE MODEL OF
CHRONIC ALLERGIC INFLAMMATION.
Marques, P. S. ; Arajo, C. C. ; Samary, C. S. ; Guimares, I. H. ; Crosseti, J. C. ; Xisto, D. G. ; Rocco, P.

R. M.
Universidade Federal do Rio de Janeiro, UFRJ / IBCCF
Objectives:
Asthma is a common disease associated with inflammatory and remodeling changes in airway and lung parenchyma. Regular and
moderate aerobic exercise has been used as an adjunct therapy to minimize the inflammatory process. However, so far, no study
has analyzed whether exercise may minimize or even reverse airway remodeling in asthma. The aim of this study was to
investigate the effects of regular and moderate aerobic exercise on the inflammatory and remodeling processes in a murine model
of chronic allergic inflammation.
28 BALB/c mice (female, 20-25g) were randomly assigned into 2 groups: in OVA group, mice were immunized by
intraperitoneal injection of ovalbumin (OVA, 10 g, ip) on 7 alternate days. After day 40, they were challenged with three
intratracheal instillations of OVA (20 g, it) at 3-days intervals. Control group (C) received saline using the same protocol.
Twenty-four hours after the last challenge animals were further subdivided into 2 groups: sedentary (S) and trained (T). T group
ran on a motorized treadmill at moderate intensity (8-12 m/min), 5% grade, 30 min/day, 3 times a week for 4 weeks. Twenty-four
hours after the last training session, lung mechanics and histology, as well as collagen fiber content in the airways were
evaluated. OVA-S group presented higher static lung elastance (63%), airway resistance (100%) and viscoelastic pressure (78%),
as well as fraction area of alveolar collapse (36%), mononuclear cells (50%), polymorphonuclear cells (37%), and collagen fiber
content (23%) compared to C-S group. OVA-T group showed a reduced static lung elastance (22%), airway resistance (65%),
viscoelastic pressure (37%), fraction area of alveolar collapse (36%), number of mononuclear cells (34%) and collagen fiber
(16%) content compared to OVA-S.
Conclusions:
In the present model of chronic allergic inflammation, regular and moderate aerobic exercise attenuated the inflammatory and
remodeling processes, improving lung mechanics.
Keywords: exercise, asthma, process inflamatory, female, remodeling
Financial Support: CNPq, FAPERJ, PRONEX, FAPERJ
Resumo:08-055
THE IMPACT OF TWO DIFFERENT SOURCES OF MESENCHYMAL STEM CELLS IN A MURINE MODEL OF
ELASTASE-INDUCED EMPHYSEMA
Antunes, M. A. 1; Abreu, S. C. 1; Cruz, F. F. 1; Oliveira, A. C. R. 1; Xisto, D. G. 1; Paz, A. H. R. 2; Cirnelima, E. O. 2; Morales, M. M. 1; Rocco, P. R. M. 1

1
Instituto de Biofsica Carlos Chagas Filho/CCS, UFRJ
2
Laboratrio de Embriologia e Diferenciao Celular, HCPA
Objectives:
Mesenchymal stem cells (MSCs) represent a promising tool for respiratory diseases. Bone marrow (BM) was the first source
reported to contain MSCs. However, for clinical use, BM may be detrimental due to the highly invasive donation procedure and
the decline in MSC number and differentiation potential with increasing age. More recently, adipose tissue (AD) was introduced
as an alternative source for MSCs. We compared the effects of MSCs derived from these sources on lung mechanics and
histology in a murine model of elastase-induced emphysema.
Thirty female C57BL/6 mice (20-25 g) were randomly assigned into 2 groups. In control (C) animals, saline was intratracheally
(it, 50 L) injected, whereas emphysema mice received porcine pancreatic elastase (ELA, 0.1 UI, 50 L, it). Saline and ELA were
intratracheally injected once a week during 4 weeks. Three hours following the last instillation, C and emphysema groups were
further randomized into subgroups receiving saline (SAL, 50 L), BM-MSC, and AD-MSC (1x105) intravenously. Adipose tissue
was harvested from mouse epididymal fat. After one week, lung mechanics (static elastance, resistive and viscoelastic pressures),
and histology (total and differential cell count in lung tissue, and fraction area of normal, collapsed and hyperinflated alveoli)
were analyzed. Lung static elastance, viscoelastic pressure, the number of polymorphonuclear cells in the tissue, and the fraction
area of alveolar hyperinflation were higher in ELA-SAL compared to C-SAL. Both BM-MSCs and AD-MSCs reduced similarly
lung static elastance (ELA-SAL: 393; ELA-BM-MSC: 313; ELA-AD-MSC: 280.5 cmH2O/ml, MeanSD; p
Conclusions:
MSCs from bone marrow and adipose tissue reduced the inflammatory process improving lung mechanics. However, at day 7 no
significant repair was observed.
Keywords: adipose, bone-marrow, elastase, emphysema, stem cell
Financial Support: PRONEX-FAPERJ, CNPq, FAPERJ, CAPES
Resumo:08-056
ESTABLISHMENT OF A CONTROLLED SWINE MODEL OF THE ACUTE RESPIRATORY DISTRESS
SYNDROME
Gomes, S. 1; Hirota, A. 1; Costa, Elv 1; Barbeiro, Df 3; Tucci, Mr 1; Beraldo, Ma 1; Nascimento, Ec2;

Belmino, R. 1; Carvalho, Crr1; Amato, Mbp 1
1
Diviso de Pneumologia - INCOR, HCFMUSP
2
Depto de Patologia, FMUSP
3
Lab de Emergncias Clnicas, FMUSP
Objectives:
The Acute Respiratory Distress Syndrome (ARDS) is defined in humans, but there is no agreement on the main characteristics of
acute lung injury in animals. The aim of this study is to establish an animal model of lung injury, which is stable and similar to
human ARDS
Methods: We tested 32 pigs (Landrace) in 4 groups (Sham n=5, Injury with histology n=7, Injury without histology n=20, and
Long term protocol ARDSNET n=10) applying a two-hit injury: first, massive lung lavage till PaO2/FiO2
Conclusions:
This model of severe ARDS mimics human ARDS, exhibiting most of its functional/inflammatory features, also presenting
stability for long periods of time.
Keywords: lung injury, experimental model, ards
Financial Support: FINEP, CNPq, CAPES e HCFMUSP
Resumo:08-057
EFFECTS OF DIFFERENT POSITIVE END EXPIRATORY PRESSURE LEVELS ON LUNG AND DISTAL ORGANS
IN MODELS OF PULMONARY AND EXTRAPULMONARY ACUTE LUNG INJURY WITH OR WITHOUT INTRAABDOMINAL HYPERTENSION.
Miranda, P. J. B. 2; Santos, C. L. 2; Oliveira, M. G. 2; Moraes, L. 2; Santos, R. S. 2; Silva, J. D. 2; Morales,

M. M. 3; Capelozzi, V. L. 4; Rocco, P. R. M. 2; Garcia, C. S. N. B. 2
2
3
Laboratory of Cellular and Molecular Physiology, IBCCF, UFRJ
4
Department of Pathology, School of Medicine, , USP
Objectives:
Pulmonary (p) and extra-pulmonary (exp) acute lung injury (ALI) differ according to their pathophysiology, even though ALI
patients undergo similar ventilatory therapy. This study investigated the impact of different positive end-expiratory pressure
(PEEP) levels in ALIp and ALIexp in the presence or absence of intra-abdominal hypertension (IAH).
Wistar rats (n=72) were randomly allocated to receive Escherichia coli lipopolysaccharide intratracheally (200 g, ALIp) or
intraperitoneally (1 mg, ALIexp). After 24 hours, they were randomized into subgroups with or without IAH. Animals developed
IAH by inserting gauze into abdominal cavity reaching a maximum stabilized intra-abdominal pressure equal to 15 mmHg. They
were then ventilated with FiO2=0.4, VT = 6 ml/kg, and PEEP = 5, 7 or 10 cmH2O during 1 hour (End). Respiratory system (rs),
lung (L) and chest wall (w) static elastances (Est), arterial blood gases, lung histology (light and electron microscopy), interleukin
(IL)-1&beta, IL-6, caspase-3 and type III procollagen (PCIII) mRNA expressions in lung tissue, as well as the number of lung,
liver, kidney and villi cell apoptosis were analyzed. Est,rs, Est,L, Est,w, PaO2, PaCO2, and pHa were comparable at Baseline in
all groups. In the presence of IAH, we found that: 1) PEEP of 5, 7 or 10 cmH2O led to oxygenation improvement independent of
ALI etiology (30%); 2) Est,w increased after surgery (38%) and remained increased until End; 3) PEEP7 led to lower Est,L and
less alveolar collapse in ALIexp, but yielded reduced alveolar-capillary membrane, epithelial and endothelial damage, cell
apoptosis in lung tissue, as well as mRNA expression of IL-1, IL-6, PCIII, and caspase-3 in both ALI groups; 4) PEEP10
resulted in hyperinflation in both ALI groups; 5) IL-1&beta, IL-6 PCIII, and caspase-3 mRNA expressions were higher only in
ALIexp ventilated with PEEP10 as well as in ALIp with PEEP5; and 6)liver, kidney, and villi cell apoptosis increased
independent of PEEP level.
Conclusions:
In the presence of IAH, PEEP7 improved lung mechanics and morphology and induced less epithelial, endothelial, and alveolar
capillary membrane damage as well as cell apoptosis in lung tissue independent of ALI etiology. However, in ALIexp, PEEP10
led to increased expression of IL-1, IL-6 PCIII, and caspase-3 in lung tissue while lower PEEP resulted in higher expression of
these mediators in ALIp. Therefore, PEEP should be set not only according to IAH but also to the etiology of ALI.
Keywords: acute lung injury, intra-abdominal hypertension, cytokines, type III procollagen, apoptosis
Financial Support: CNPq, PRONEX-FAPERJ, FAPERJ, CAPES
Resumo:08-058
ACTIVITY OF THE COMPOUNDS IQG-607 AND IQG-639 IN A MURINE MODEL OF TUBERCULOSIS
Rodrigues-junior, V. S. 1,2; Santos, A. A. Jr. 1,3; Jader, A. S. 1; Ritter, E. 1; Souto, A. A. 1; Calixto, J. B. 4,1;
Basso, L. A. 1,3; Santos, D. S. 1,3; Campos, M. M. 1,2,5
1
Instituto Nacional de Cincia e Tecnologia em Tuberculose, PUC RS
5
Instituto de Toxicologia, InTox, PUC RS
2
Programa de Ps-Graduao em Medicina e Cincias da Sade, PPGMCS, PUC RS
3
Programa de Ps-Graduao em Biologia Celular e Molecular, PPGBCM, PUC RS
4
Departamento de Farmacologia, UFSC
Objectives:
Tuberculosis (TB) continues to be one of the deadliest diseases in the world. The emergence of multi-drug-resistant strains of
Mycobacterium tuberculosis, the unbearable side effects of the available drugs, and the frequent patient non-compliance in
completing the therapy have increased the need for development of new effective agents. Isoniazid (INH) is the most prescribed
drug for active TB and prophylaxis, and requires activation by the enzyme KatG. The M. tuberculosis enoyl-ACP reductase
(InhA) has been shown to be the primary target for INH. Our group has published the rational design and synthesis of new
isoniazid-derived compounds with possible anti-TB activity. Importantly, it was shown that these compounds do not require
activation by KatG to bind its molecular target InhA. We have also demonstrated that INH analogs are able to inhibit the activity
of wild-type and INH-resistant M. tuberculosis InhA in vitro. This work describes the pre-clinical in vivo evaluation of two
pentacyano(isoniazid)ferrateII complexes, named IQG-607 and IQG-639, in a mouse model of TB.
All the experimental protocols were approved by the local Animal Ethics Committee (CEUA 09/00094). Male Swiss mice (N=5,
25-30 g) were intravenously infected, with 1 x 107 viable M. tuberculosis bacilli suspended in 0.2 ml of Middlebrook 7H9
medium. Treatment was started 5 days post-infection, and the compounds IQG-607 and IQG-639 (250 mg/kg) were administered
orally, once a day, during 28 or 56 days. Separate groups of mice were treated with the reference drug INH (25 mg/kg; positive
control) or saline solution (negative control). At the end of treatments, mice were euthanized by isoflurane inhalation. The
spleens and lungs were aseptically removed, and the spleen weight (in grams) was determined. These organs were homogenized
and the colony-forming units (CFU) were determined after 30 days of incubation in solid medium, at 37 C. After 28 days of
treatment, either IQG-607 or INH significantly reduced M. tuberculosis-induced splenomegaly, with inhibition percentages of 50
and 58 %, respectively. Interestingly, after 56 days of treatment, the reduction of the splenomegaly by either IQG-607 or INH
was increased, with inhibition percentages of 67 and 66 %, respectively. Moreover, after 28 or 56 days of treatment, CFU
numbers were significantly reduced in either spleens or lungs of IQG-607- and INH-treated groups. Otherwise, IQG-639 was not
capable of significantly modifying any evaluated parameter.
Conclusions:
The promising activity of IQG-607 in M. tuberculosis-infected mice suggests that it is a good candidate for clinical development
as a new anti-tuberculosis agent. Additional experiments are being currently performed to determine the pharmacokinetic and
toxicological parameters of this compound.
Keywords: Tuberculosis, InhA inhibitors, Preclinical Test, Drug Development
Financial Support: INCT-TB CNPq, BNDES, FINEP, CAPES
Resumo:08-059
ELECTROPHYSIOLOGICAL STUDIES OF THE DIAPHRAGM OF ELASTASE-INDUCED EMPHYSEMA IN
MICE.
Cndido, S. C. O; Teodoro, L. C. ; Zavan, B. ; Bento, A. C. ; Paiva, A. G. ; Junior, V. A. P. ; Soncini, R.

Cincias Biomdicas - Universidade Federal de Alfenas, UNIFAL - MG
Objectives:
Emphysema, a type of obstructive disease (COPD), has morphological and functional changes of the whole lung tissue. The
changes are generally in respiratory mechanics, increased air spaces, alteration of histological parameters and airway extracellular
matrix of the. The aim of this study was to evaluate electrophysiological and histochemical and imunohistochemical changes of
the diaphragm of elastase-induced emphysema in mice.
Materials and methods: Thirty-two Swiss male mice (25-30g) were randomly divided into control group and emphysema. The
control group received intratracheal instillation of saline, single dose (Sal x 1) and double dose (Sal x 2). The emphysema group
received intratracheal instillation of porcine pancreatic elastase, dose 0,3 U/mouse, single dose (0,3 U x 1) and double dose (0,3
U x 2). Twenty days after the first instillation, the mice were decapitated to isolate from the left hemi-diaphragm fragment of 5
mm width . The fragments were suspended in an isolated organ bath filled with Krebs with d-tubocurarine. After 30 min, was
started the electrical stimulation to construct the curve force-frequency, using a voltage to maximal stimulus. Power Lab system
was used to register the Force (g). For morphological analysis, the diaphragm and lungs were removed and embedded in paraffin.
In the study of the diaphragm was used picrosirius for colagen fibers and hematoxylin eosin for immunocytochemistry of actin. The lungs were prepared for histology and morphometry. For statistical analysis, the results were expressed as mean
SEM. For comparison of two means was applied to t-test of Student. For comparison of three or more means applied - the
analysis of variance (One-way) followed by Tukey post-test when appropriate. The significance level for rejection of the null
hypothesis was set at 5% (p
Conclusions:
Conclusion: The data show that the strength of the diaphragm is not altered in emphysema group, indicating that its function is
preserved. However, the morphology of the diaphragm indicates that there is increased collagen and actin. Then, the results
indicate that there may be a relationship between morphological settings and preservation of function of the diaphragm of
elastase-induced emphysema in mice.
Keywords: collagens fibers, Diaphragm, Elastase, Mice, Pulmonary Emphysema
Resumo:08-060
COMPARISON OF TWO DIFFERENT ROUTES OF DELIVERING ADIPOSE-DERIVED STEM CELLS IN A
MURINE MODEL OF EMPHYSEMA.
Oliveira, A. C. R. D. 1; Antunes, M. A. 1; Abreu, S. C. 1; Cruz, F. F. 1; Xisto, D. G. 1,2; Morales, M. M. 1,2;

Rocco, P. R. M. 1,2
1
Laboratrio de Investigao Pulmonar - IBCCF, UFRJ
Laboratrio de Fisiologia Celular e Molecular - IBCCF, UFRJ
Objectives:
To investigate the effects of two different routes of adipose-derived stem cell (ASC) administration on lung mechanics and
histology in a murine model of elastase-induced emphysema.
Thirty female C57BL/6 mice were randomly divided into control (C) and emphysema (E) groups. E groups were intratracheally
instilled with 0.1U of porcine pancreatic elastase (PPE) in 50 L of saline once a week during four weeks, and C groups received
only 50 L of saline per instillation. Three hours following the last instillation, saline or ASC, which was obtained from
epididymis of male mice, were intravenously (i.v.) or intratracheally (i.t.) administrated in C (C-ASC i.v. and C-ASC i.t.) and E
(E-ASC i.v. and E-ASC i.t.) groups. After one week, lung static elastance (Est), resistive (P1) and viscoelastic (P2) pressures,
total and differential cell count in lung tissue, and fraction area of normal, collapsed and hyperinflated alveoli were analyzed. ESAL group presented higher Est (+23.1%), P2 (+11.2%), and fraction area of alveolar hyperinflation (23.9 3.3%) compared to
C-SAL group. ASC reduced lung elastance (E-ASC i.v.: -28.6% and E-ASC i.t.: -26.6%) and viscoelastic (E-ASC i.v.: -28.8%
and E-ASC i.t.: -24.7%) similarly independent of administration route. However, ASC was unable to revert hyperinflated areas,
but was effective in reducing inflammatory cell infiltration in lung tissue in both ASC routes.
Conclusions:
ASC modulates the inflammatory processes, improving lung mechanics in the present murine model of elastase-induce
emphysema.
Keywords: DIFFERENT ROUTES, ADIPOSE-DERIVED STEM CELLS, EMPHYSEMA
Financial Support: FAPERJ, CNPq, CAPES, PRONEX
Resumo:08-061
APOCYNIN AND TEMPOL PROMOTE INCREASE OF NO PRODUCTION IN OVALBUMIN-INDUCED ASTHMA
IN BALB/C MICE.
Nesi, R. T. 1; Lanzetti, M. 2,2; Santos-silva, M. A. 2; Pires, K. M. 2; Guilherme, R. 1; Neves, J. S. 1;

Benjamim, C. F. 1; Valena, S. D. S. 1
1
FEDERAL UNIVERSITY OF RIO DE JANEIRO , UFRJ
2
State University of Rio de Janeiro, UERJ
Objectives:
The events that contribute to both oxidative and nitrosative stresses in asthma are well described. However, the effects of
superoxide on oxidative damage and NO production in asthma are poorly understood. The present study investigated the role of
superoxide in ovalbumin-induced asthma.
Male BALB/c mice were assigned into six groups: Control (CTR), CTR tempol, CTR apocynin, ovalbumin (OVA), OVA tempol
and OVA apocynin group (n=30; five animals per group). The CTR groups were sensitized with 200 L of phosphate buffer and
the OVA groups were sensitized with 200 L of solution containing 50g of OVA and 5 mg of adjuvant. All of animals were
sensitizated subcutaneously on 0 day and through intraperitoneal injection (i.p.) on 14st day. After 4 days, both groups were
challenged with 2% nebulized OVA (30 minutes/day for 3 consecutive days). Before each challenge, the animals from CTR
tempol, CTR apocynin, OVA tempol and OVA apocynin were submitted to administration of dismutase superoxide (SOD)
mimetic called tempol (100mg/Kg weight) and NADPH oxidase inhibitor known as apocynin (14mg/Kg weight) by i.p. On the
23st day each animal was sacrificed and the half of the lung was homogenized and the other half was fixed for histological
analysis and bronchoalveolar lavage (BALF) was done for the following biochemical dosages: the total content of ROS; catalase
(CAT), eosinophil peroxidase (EPO) activities; nitrite levels; hydroperoxides levels. The total content of ROS was increased in
OVA group (0.26 0.08) when compared to CTR group (0.03 0.01; p
Conclusions:
Both drugs promoted the increased of NO production OVA-induced asthma. We suggest that, although superoxide pathway may
not influence oxidative damage, NO production may contribute to nitrosative damage in experimental asthma.
Keywords: superoxide, nitric oxide, asthma, apocynin, tempol
Financial Support: CNPQ, Capes and FAPERJ
Resumo:08-062
BONE MARROW-DERIVED MONONUCLEAR CELL THERAPY MODULATES THE INFLAMMATORY AND
REMODELING PROCESSES INDEPENDENT OF THE ADMINISTRATION ROUTE IN EXPERIMENTAL
CHRONIC ALLERGIC ASTHMA
Abreu, S. C. 1; Antunes, M. A. 1; Cruz, F. F. 1; Maron-gutierrez, T. 1,2; Ornellas, D. S. 1,2; Teodoro, W. R. 3;

Capelozzi, V. L. 3; Morales, M. M. 2; Rocco, P. R. M. 1
1
Laboratrio de Investigao Pulmonar - IBCCF/UFRJ, UFRJ
2
Laboratrio de Fisiologia Celular e Molecular - IIBCCF/UFRJ, UFRJ
3
Departamento de patologia, Universidade de So Paulo, USP
Objectives:
This study investigated the best administration route of bone marrow-derived mononuclear cell in a murine model of chronic
allergic asthma, based on the analysis of the inflammatory and remodeling processes.
Thirty-six female C57BL/6 mice were randomly assigned into six groups. In the OVA group, mice were sensitized and
challenged with ovalbumin while control group (C) received saline using the same protocol. C and OVA groups were further
randomized into subgroups receiving saline (50 L, SAL) or bone marrow-derived mononuclear cell (BMDMC, 2x106, CELL)
intravenously or intratracheally, 24 hours before the first challenge. Airway and lung parenchyma inflammation and remodeling
were evaluated using light, confocal and electron microscopy. In addition, airway resistance, viscoelastic pressure, static
elastance were analyzed. BMDMC therapy led to a reduction, independent of the route of BMDMC administration, in alveolar
collapse, eosinophil infiltration, broncoconstriction index, smooth-muscle actin expression, subepithelial fibrosis, myocyte
hypertrophy and hyperplasia, and the amount of myofibroblasts in airways and lung parenchyma compared to OVA-SAL. Airway
resistance and viscoelastic pressure were significantly more reduced after cell therapy with intratracheal instillation (38% and
24%, respectively) compared to intravenous (40% and 25%).
Conclusions:
In the present model of chronic allergic asthma, BMDMC therapy was effective at modulating the inflammatory and fibrogenic
processes independent of the route of cell administration.
Keywords: asthma, remodeling , stem cells, administration route
Financial Support: PRONEX-FAPERJ, CNPq, FAPERJ, CAPES, INCT-INOFAR.
Resumo:08-063
DOES CHRONIC INTERMITTENT HYPOXIA PRODUCE CHANGES IN THE RESPIRATORY CONTROL OF
UPPER AIRWAY RESISTANCE? CELLULAR AND SYSTEMIC EVIDENCES.
Moraes, D. J. A. ; Bonagamba, L. G. H. ; Zoccal, D. B. ; Machado, B. H.

Dpto of Physiology/School of Medicine of Ribeiro Preto, FMRP/USP
Objectives:
Introduction: During eupneic breathing, the control of upper airway resistance is characterized by pre-inspiratory (pre-I)
abduction and post-inspiratory (post-I) adduction causing decreases during inspiration and increases during expiration
respectively. This dynamic coordination by central mechanisms over the breathing cycle has been suggested to be vital for the
maintenance of blood gases homeostasis in various respiratory-related behaviors, including during conditions of hypoxia. Aims:
To evaluate if chronic intermittent hypoxia (CIH) produces changes in the respiratory control of upper airway resistance in the
awake rats and in the working heart-brainstem preparation (WHBP) and to characterize the electrophysiological mechanisms
involved in these changes.
CIH paradigm consisted of 6% O2 for 40 s every 9 min, 8 h/day. On the 11th day, the EMG of diaphragm (DIA) and tongue
protrudor genioglossus (GG) muscles were evaluated simultaneous with plethysmographic recordings in awake rats breathing
room air. In the WHBP, phrenic (PN), cervical vagus (cVN), hypoglossal (HN) and recurrent laryngeal (RLN) nerves activities
were recorded. In addition, intracellular recordings of pre-I and post-I neurons of pre-Btzinger (pre-BtC) and Btzinger
Complex (BtC) were made in presence or not in presence or not of antagonists of excitatory and inhibitory synaptic
neurotransmission (KYN: 1-6 mM; Biccuculine: 20 M and Strychnine: 1M). By these recordings, the membrane input
resistance and the neuronal excitability (the threshold current required to evoke an action potential) were determined. In awake
CIH rats (n=7) the pre-I abduction of GG was greater than in control (n=4) (153 vs 20.1%; p<0.05).
Conclusions:
We suggest that the CIH reduces upper airway resistance in unanesthetized rats during inspiration and expiration, by mechanisms
involving electrophysiological changes in the BtC post-I and pre-BtC pre-I neurons.
Keywords: Chronic Intermittent Hypoxia, Respiratory Control, Upper Airway Resistance, Btzinger Complex, pre-Btzinger
Complex
Financial Support: CAPES, CNPQ and FAPESP
Resumo:09-026
EFFECT OF THE ESTROGEN IN MODULATION OF THE HEXOKINASE MITOCHONDRIAL AS POSSIBLE
ANTIOXIDANT PREVENTIVE AGENT OF THE ROS GENERATION
Ferreira, C. R. ; Filho, A. G.
Objectives:
The brain is a tissue that demands of lot of energy, being the mitochondrial oxidative phosphorylation essential to the neurons to
maintain the high demands of ATP. In addition, in situations of bioenergetic crisis, the decrease mitochondrial respiration may
produce reactive oxygen species (ROS) generation. Among many mechanisms suggested in literature to the decrease of
mitochondrial ROS, the estrogen appears as the physiological sex-specific regulator of ROS production. Estrogen induces
association of the Hexokinase-II (HK-II) to voltage dependent anionic channel (VDAC) in neuron cells. Our group had
demostraded that rat and mice brain mitochondrial hexokinase (HK) activity play a critical role ADP/ATP re-cycling decreasing
ROS production (J Biol Chem., 38, 39846, 2004). The aim of this study was to analyze in rats whether the gender and ages could
affect the ROS generation and HK activities in rat brain mitochondria. In addition ovarioctemized rats were used in order to
challenge the hypothesis of estrogen prevention of ROS formation.
The brain mitochondria were isolated of rats (200-350 g) by percoll gradient. The detailed analyses of ETC by oxygen uptake
were performed using high-resolution respirometry (Oroboros Oxygraph-O2K). The ROS production was measured by
fluorescence method Amplex Red. The results showed that when males e females (4 months age) were compared for respiratory
substrates (2 mM pyruvate/malate, 5 mM glutamate and 10 mM succinate) no differences were observed.. There was a trend to
stimulate more respiration by activating of the HK with addition of 20 mM 2-DOG in females than in males. No differences were
observed in stimulated ROS generation in both males and females. However, in females younger (2 months) were observed
differences in the function mitochondrial of, ATP synthesis (state 3) and stimulated respiration by HK activation using 2-DOG
increases by 1.5 folds when compared to females older (4 months). Despite of these observations, sham operated rats presented
bigger respiration in state 3, induced by 2-DOG and FCCP than ovarioctemized rats of same age (2 months). In addition, the
respiration of state 3 and FCCP (maximum respiration) were almost the same in younger females. Whereas, in older females, the
respiration induced by state 3 were only 1/3 of that induced FCCP. The ROS production showed a tendency to be higher in
ovarioctemizades than sham rats.
Conclusions:
Our data suggested that estrogens could play a role modulating HK contributing to decrease ROS production. Female rats of the
different ages showed different respiratory profiles and ROS production. Despite of apparent lack differences in rats older of the
different sex, more studies are necessary to a better evaluation of estrogen effects in brain mitochondria.
Keywords: Electron transport chain, Estrogen, Hexokinase, Mitochondria, Species oxygen reactive
Financial Support: Capes, CNPq, FAPERJ, INCTEN
Resumo:09-027
CROSS-TALK BETWEEN G-PROTEIN-COUPLED AND P2X RECEPTORS IN MICE LEYDIG CELLS
Costa, R. R. ; Aguiar, J. F. ; Varanda, W. A.

Depto. Fisiologia- Faculdade de Medicina , FMRP-USP
Objectives:
Ca2+ influx and intracellular Ca2+ increase are essential to testosterone production and secretion in Leydig cells. In these cells Ttype Ca2+ channels are activated by luteinizing hormone (LH) and are modulated by protein kinases. In adittion to T-type
channels, P2X receptors are also present in the plasma membrane of Leydig cells and may serve as another pathway for Ca2+
entry. Consistent with this ATP effectively increases testosterone secretion. As LH leads to the activation of adenylate ciclase and
phospholipase C, increasing intracellular Ca2+ concentration [Ca2+]i, our goal was to investigate the effects of LH on ATP
currents and on [Ca2+]i stimulated by ATP.
Whole-cell currents were measured in Leydig cells in response to 100 M ATP before and after LH and H89 (blocker of PKA).
Fluo-3 and mag-fluo4 were used in order to simultaneously monitor the cytosolic and endoplasmic reticulum Ca2+ activity,
respectively. Fluorescence was measured on a Leica SP-5 confocal microscope. We have found that inhibition of PKA
significantly reduced the amplitude of ATP activated currents at -60 mV, from -179.7 9 to -89.4 25 pA (n= 13 cells) (H89
applied 5 min before testing the ATP). However, application of LH between ATP pulses did not alter the ATP activated currents.
ATP (100 M), induced a fast 3.7 0.1 fold increase in fluorescence in relation to basal level, that did not occur in the absence of
external Ca2+ or in the presence of 300 M suramine This response was significantly reduced by 1.9 0.1 fold by the PKA
inhibitor H89 (1 M) and by 2.5 0.2 fold following a 5 minutes incubation with LH. Ryanodine (100 M) did not modify the
amplitude and the time to peak of the [Ca2+]i transients induced by ATP, but the half-duration (1/2D) is significantly larger.
Interestingly, a previous stimulation of the cell with ATP impaired the [Ca2+]i typically induced by a subsequent application of
LH. A line scan analysis of the fluorescence indicated that the increase in [Ca2+]i starts close to the plasma membrane.
Simultaneously with the increase in [Ca2+]i stimulated by ATP on the cytosol there is a depletion of Ca2+ in the endoplasmic
reticulum. Comparing the fluorescence transients we see larger amplitudes in the case of ATP ( 3.6 0.1) than with LH (2.2
0.2). The time to peak is faster for ATP (9.2 0.4 s) than for LH (14 1.1 s), and the half-duration is shorter for LH (14.3 1.8
s) than for ATP (25 1.7 s).
Conclusions:
In these experiments ATP activated cation currents were reduced by PKA inhibition, but are not modified by LH. In terms of
Ca2+ signaling, we confirmed the presence of P2XR and we observed that RYR inhibition did not abolish the purinergic Ca2+
transient. In contrast, the Ca2+ transients in response to ATP were reduced by the inhibition of PKA. This suggests a possible
role of PKA as modulator of the purinergic induced Ca2+ transients. These results also imply that ATP-mediated P2X receptor
activity exerts an inhibitory influence on the final effect of LH on Leydig cells and therefore on testosterone secretion.
Keywords: Leydig cells , P2x receptorts, calcium signaling
Financial Support: FAPESP, FAEPA
Resumo:09-028
EFFECT OF EBSELEN ON MICE BRAIN MITOCHONDRIA METABOLISM
Pereira, P. R. P. ; Camacho-pereira, J. ; Galina, A.

Objectives:
Ebselen (EBS), also called PZ51, is a selenium drug with a variety of pharmacological properties and therapeutic which have
been associated as a mimetic of glutathione peroxidase and thioredoxin reductase . It has been demonstrated that EBS protect
against deterioration of brain function in patients with cerebral infarction and subarachnoid hemorrhage. EBS underwent several
clinical trials and is now considered as a potential antioxidant drug to treat diseases associated with excessive oxidative stress, but
further studies are needed to establish the threshold doses that decrease reactive oxygen species (ROS) formation without causing
further cellular stresses. In this paper, we evaluate the effect of EBS in a central enzyme of glucose metabolism, hexokinase, and
the antioxidant effect of EBS in mitochondria ROS generation from mice brain.
EBS was able to inhibit a key metabolic enzyme such as hexokinase, which phosphorylates glucose to glucose-6-phosphate.
Measuring the activity of hexokinase by spectrophotometric assay in isolated mitochondria of mouse brain, we observed that their
activity was reduced by almost 50% with 5M EBS. It was evaluated the effect of EBS on mitochondrial generation of ROS
measured with the Amplex Red oxidation method. We observed that EBS has a biphasic effect in which with EBS in the range of
1 M to 3 M there was an increase in brain mitochondria ROS production and above 5 M until 10 M, EBS promoted a
reduction in ROS formation.
Conclusions:
It was found that EBS is able to inhibit the hexokinase activity, an enzyme that is involved in mitochondrial ROS homeostasis. In
addition, EBS has a dual effect depending on the dose of antioxidant. We do not know whether both effects are correlated. These
data suggest that EBS could be an inhibitor of cell glucose metabolism, even at low doses.This data is important in relation to the
compound toxicity might cause when used for clinical treatment.
Keywords: BRAIN, EBSELEN, METABOLISM, MITOCHONDRIA
Financial Support: CNPQ FAPERJ
Resumo:09-029
REGULATION OF WILSON DISEASE ATPASE (ATP7B) ACTIVITY BY PROTEIN KINASE C (PKC).
Cardoso, L. H. D. ; Hilrio-souza, E. ; Vieyra, A. ; Lowe, J.

Instituto de Biofsica Carlos Chagas Filho, UFRJ, IBCCF
Objectives:
Copper is an essential metal in all organisms, although in high concentrations it becomes toxic. Therefore, copper level in the
organisms must be highly regulated. ATP7B is a copper transporting ATPase expressed in liver that is essential in copper
homeostasis, and it is known that Wilson disease is caused by mutations in this protein. In previous studies we have already
shown that its ATPase activity is inhibited when phosphorylated by protein kinase A (Int. J. Biochem. Cell Biol. 43: 358, 2010).
In the present work we investigated whether protein kinase C signaling pathway might also modulate this Cu(I)-ATPase activity.
In a porcine liver preparation (J. Biol. Chem. 267: 12016, 1992), different isoforms of protein kinase C (, , ) were detected by
Western Blotting. A colorimetric assay was used to detect Cu(I)-ATPase activity in the preparation (J. Biol. Chem. 66: 375,
1925), and the results were given in nmol x mg-1 x min-1 (means SE). Phorbol 12-myristate-13-acetate (PMA), a PKC
activator, increased ATP7B activity (control: 16.95 0.86; 10 nM PMA: 26.44 1.84; n4), while calphostin C, a PKC inhibitor,
decreased its activity (control: 36.94 0.54; 10 nM calphostin C: 23.21 6.76; n6). Addition of phosphatase after preincubation with PMA also decreased its activity to the same level of addition of phosphatase without PMA (control: 22.84
1.07; phosphatase : 10.07 3.00; 10 nM PMA /phosphatase : 10.25 2.38; n4). No changes were observed when Ca2+ 2 M
or PMA/Ca2+ 2 M were added, however PMA/EGTA increased the Cu(I)-ATPase activity (control: 17.47 1.71; 10 nM PMA
/Ca2+ 2 M: 19.11 2.72; Ca2+ 2 M: 21.49 2.30; 10 nM PMA/ 2 mM EGTA: 35.70 1.97; n6), indicating the involvement
of a PKC isoform which is not stimulated by calcium. The activation of the Cu(I)-ATPase activity is reverted by the PKC
specific inhibitor (control: 30.66 1.69; 0.5 M iPKC: 12.24 0.59; 0.5 M iPKC: 26.77 2.05; n4), showing that this novel
PKC isoform is responsible for the stimulation of ATP7B activity.
Conclusions:
ATP7B activity from porcine liver is regulated by PKC, which leads to a stimulation of the copper transporting activity.
Different signaling pathways involving phospholipase C in hepatocytes can activate PKC, which in turn could increase ATP7B
activity and therefore be an important element for copper homeostasis.
Keywords: ATP7B, PKC, Wilson disease, protein kinases
Financial Support: FAPERJ, CNPq, PIBIC-CNPq.
Resumo:09-030
CHARACTERIZATION OF THE ACTIVITY OF NA+-ATPASE IN HUMAN BREAST CANCER CELLS MCF-7
Dias, M. A. ; Fonseca, M. M. ; Capella, M. A. M. ; Lopes, A. G.

Objectives:
Recently, the ouabain insensitive, K+-independent Na+-ATPase was identied, isolated and characterized in guinea-pig small
intestinal cells (Biochim. Biophys. Acta (Biomembranes) 1808; 1684, 2011). This protein was previously characterized in a
number of kidney and intestinal cells, but there is no study on its expression or activity in tumor cells. The objective of this study
was to characterize the presence of Na+-ATPase activity in human breast cancer MCF-7 cells, comparing with that already
described for MDCK cells.
To observe the sensitivity of MCF-7 and MDCK cells to furosemide, the best Na+-ATPase inhibitor known, the cells were
seeded in 96 wells plates and incubated for 48 h with furosemide concentrations varying from 100 nM to 2 mM and the cellular
viability was measured by MTT. Both cell lines were sensitive to furosemide concentrations above 250 uM (p
Conclusions:
MCF-7 cells present a very low, undetected activity of Na+-ATPase in basal condition which can be increased after exposure to
hormones related to body volume regulation, suggesting that altered body volume (for example, hypertension) may alter the
expression/activity of this protein in tumor cells.
Keywords: Na+-ATPase, Breast Cancer, Ionic Transporters, MCF-7 Cells
Financial Support: CNPq, CAPES, FAPERJ, PRONEX
Resumo:09-031
REGULATION OF SODIUM INDEPENDENT GLUTAMATE TRANSPORT BY HYDROCORTISONE IN
CULTURED CHICK RETINAL CELLS
Garza, . D. R. 1; Oliveira, K. R. M. 1; S, L. L. C. 2; Garcia, T. B. 1; Crespo-lopez, M. H. C. 1; Herculano, A.

M. 1
1
Universidade Federal do Par , UFPA/ICB
2
Instituto Evandro Chagas - Seo de Meio Ambiente, IEC
Objectives:
Aim: Transport of glutamate in the vertebrate central nervous system is crucial to assure a high signal-to-noise synaptic
transmission and to prevent excitotoxicity. In retinal tissue a family of sodium dependent and independent transporters are
responsible for the majority of the uptake of glutamate released at the synaptic cleft. Recently we demonstrated that about 40% of
the glutamate transport in retinal tissue is mediated by sodium-independent transporters (Neurochem int. 56:59, 2010). The aim
of the present study is to evaluate the neuroedocrine regulation of the sodium independent glutamate/cysteine (-SH) transport in
retinal cells. Hydrocortisone was used in our analysis since previous reports have demonstrated its involvement in the regulation
of two glial proteins crucial for the glutamate transport phenotype: the sodium-dependent transporter EAAT1 (Neurochem int.
37:179, 2000) and the cytoplasmatic Glutamine Sinthethase that cycles glutamate to glutamine (Proc. Natl. Acad. Sci USA.
91:4786, 1994). Thus, using primary retinal cell cultures as experimental model, we investigated if hydrocortisone is able to
regulate the sodium independent transport of glutamate in the retina.
Methods and Results: primary cultures where established from retinal cells of E8 chick embryos and cultivated at 37C with a
humid 5% CO atmosphere in a DMEM medium containing 10% FBS for 7 days. Glutamate transport was evaluated by high
performance liquid chromatography as described previously. Extracellular SH content was determinated by DTNB reduction.
The experiments in medium without sodium, NaCl was replaced by equimolar concentration of LiCl. Our results have
demontrated that retinal cell cultures treated with hydrocortisone (0,33 g/ml) showed a significant increase of glutamate uptake
(39 5,88% over the control). Similar increase was observed in the cultures incubated in a sodium medium free ( 29 6,7% over
control). This result suggests that hydrocortisone evokes a glutamate uptake mediated by sodium independent transporter. To
ratify this hypothesis we evaluate the effect of hydrocortisone in the SH release in a medium without sodium as described
above. Our data showed an increase of SH content in a sodium-independent manner (76,81 5,57% over control).
Conclusions:
Conclusions: Hydrocortisone treatment induces an increase in the glutamate uptake mediated by sodium independent glutamate
transporters and this effect was followed by an intense release of SH in the medium suggesting a possible role of
glutamate/cystine antiporter. hydrocortisone treatment of chick retinal cultured cells increases the total extracellular sulfidryl
when stimulated with L-glutamate in a sodium-independent manner and increases the uptake of glutamate in both sodium
independent and dependent systems.
Keywords: Cystine/glutamate antiporter, Glutamate uptake, Hydrocortisone, Retina, Sulfidryl
Financial Support: CNPQ/ FAPESPA/MAKAR
Resumo:09-032
CHANGES IN BROWN ADIPOSE TISSUE THERMOGENESIS INDUCED BY THYROID HORMONE
Curty G. 1; Salvatte, K. 1; Carvalho, D. P. 2; de Meis, L. 1; Ketzer, L. A. 1,3

1
INSTITUTO DE BIOQUMICA MDICA, RIO DE JANEIRO , UFRJ
2
INSTITUTO DE BIOFSICA CARLOS CHAGAS FILHO, RIO DE JANEIRO, UFRJ
3
PLO AVANADO DE XERM, RIO DE JANEIRO / XERM, UFRJ
Objectives:
Homeothermic animals control body temperature by using endogenous mechanisms to produce and dissipate heat. These
mechanisms comprise thermogenesis, which can be divided into obligatory or adaptive (facultative). The obligatory
thermogenesis is the basal metabolic rate and occurs when the organism is at rest. Already the adaptive thermogenesis occurs in
response to diet and environmental factors as cold exposure. Thyroid hormones increase the basal metabolic rate and are
considered essential for adaptive thermogenesis. The skeletal muscle and brown adipose tissue (BAT) are important sites for
adaptive thermogenesis. It is suggested that the adaptive response in muscle is promoted by shivering and the activity of some
enzymes, such as sarco(endo)plasmic reticulum Ca2+-ATPase(SERCA 1).In skeletal muscle, SERCA 1 is an enzyme able to
pump Ca2+ from the cytosol into the sarcoplasmic reticulum lumen using the energy derived from ATP hydrolysis. However,
SERCA 1 is able to hydrolyze ATP without pumping Ca2+, producing more heat per ATP cleaved than that released during Ca2+
transport. In the BAT, the uncoupling protein 1 (UCP 1) contributes significantly to adaptive thermogenesis. Recently was
identified the expression of SERCA 1 in the mitochondria and endoplasmic reticulum of BAT through immunoelectron
microscopy. However, the role of this enzyme in this tissue is not clear. The purpose of this study is to investigate the
thermogenic role of SERCA 1 in BAT in response to increased thyroid hormone, as well as expression and activity of UCP1 in
these animals.
Hyperthyroidism was induced in Wistar rats by injection of T4 (100 g/kg body weight), subcutaneously, for 10 days.
Hyperthyroid rats had a decrease of body weight gain (29.6 3.5 g versus 40.1 4.2 g in control, n=8) and an increase of BAT
weight (56 %). The endoplasmic reticulum vesicles from BAT were prepared by differential centrifugation and stored at 70C.
The mitochondrial fraction was obtained by Percoll gradient. The thermogenic parameters were measured using an isothermal
titration calorimeter and high-resolution oxygraph. Thyroid hormone induced SERCA 1 expression and promoted an increase in
heat released by ATP hydrolysis in the endoplasmic reticulum, suggesting an increase in thermogenesis. Citrate synthase activity,
an enzyme of the Krebs cycle, was used as an indicator of mitochondrial biogenesis. Although the citrate synthase activity to
show an increase in hyperthyroidism (1.79 0.2 mol/mg.min versus 1.26 0.2 mol/mg.min in control), the UCP1 expression
was unchanged. Hyperthyroidism promoted a rise in oxygen consumption (0.65 0.1 mol O2/mg.min versus 0.3 0.05 mol
O2/mg.min in control) and heat production (7 fold) by mitochondria.
Conclusions:
These data suggest that thyroid hormones modulate not only the UCP1 activity, but also the SERCA 1 expression and the activity
of in BAT. Here we show the importance of the thyroid hormones in adaptive thermogenesis promoted by BAT.
Keywords: Brown adipose tissue, Thermogenesis, Thyroid hormones, Ca2+-ATPase of sarcoplasmic reticulum, uncoupling
protein 1
Financial Support: CNPq, FAPERJ, FINEP
Resumo:09-033
VALINOMYCIN AND OUABAIN PREVENT VERAPAMIL INHIBITORY EFFECT ON EMBRIONIC
DEVELOPMENT OF THE SEA URCHIN ECHINOMETRA LUCUNTER.
Leite, J. C. A. ; Marques-santos, L. F.
Deptartamento de Biologia Molecular, UFPB
Objectives:
Ca2+ is an intracellular messenger that controls a wide range of physiological functions. The increase in cytosolic calcium
concentration is derived from intracellular stores mobilization, e.g. endoplasmatic reticulum (ER), or influx through channels,
especially voltagegated channels (Cav) present on cell surface. According to scientific literature, sea urchin embryogenesis is a
process induced exclusively by Ca2+ mobilization from ER, as Ca2+ influx is not necessary for this process (PNAS 75:1915,
1974). However, our group previously showed that verapamil (VP), an unspecific calcium channel blocker, induced a significant
blockage on early embryonic development of the sea urchin Echinometra lucunter. Similar results were obtained with diltiazem
and nifedipine. These data suggest calcium influx dependence on embryogenesis of this species. In the present work we
investigate the effect of the potassium ionophore valinomycin (VAL) and the Na+K+ ATPase inhibitor ouabain (OUA) on VP
inhibitory effect over E. lucunter early embryonic development. K+ efflux mediated by VAL alters membrane potential, which
can influence the opening of some types of Cav and its pharmacological regulation (J. Cell Sci. 98: 343, 1991). Alternatively,
Na+K+ ATPase inhibition by OUA activates the reverse mode of the Na+Ca2+ exchanger, promoting Ca2+ influx (J.
Pharmacol. Sci. 110: 111,2009).
Animals were collected at Cabo Branco Beach (Joo Pessoa, Brazil). Eggs and spermatozoa were collected by KCl intracoelomic
injection. Fertilization was induced by the addition of activated sperm suspension to eggs suspension (1:5000). Embryos were
treated with the VAL or OUA, before or after VP (100 M) treatment. One hundred embryos were analyzed to each treatment
and early embryonic development (24 cell embryo and morula) was monitored under optical microscopy. Pretreatment with
VAL and OUA blocked VP inhibitory effect. Although, VAL and OUA did not prevent VP inhibitory effect when added after the
calcium channel blocker.
Conclusions:
This set of data strongly suggests that VP inhibitory effect on E. lucunter embryonic development was induced by the impairment
of Ca2+ influx through Cav and this influx is crucial in the first minutes of the embryogenesis. Our work highlighted the
involvement of extracellular Ca2+ on embryonic development of this specie.
Keywords: Verapamil, Valinomycin, Ouabain , Embrionic Development, Echinometra lucunter
Financial Support: CNPq /CAPES
Resumo:09-034
ELECTROTHERAPY STUDY OF THE HEALING PROCESS IN ANIMALS: THE USE OF SODIUM DICLOFENACIONTOPHORESIS ADMINISTRATION
Xavier, A. F. 1; Medeiros, M. A. M. B. 2; Nascimento, R. F. 2; Oliveira, J. M. 3; Silva, B. A. 2; Souza, E. P. 2

3
Faculdades Integradas do Cear, FIC
2
Universidade Federal da Paraba, UFPB
1
Universidade Federal do Cear, UFC
Objectives:
Iontophoresis is a noninvasive technique that uses a power (
We used 20 animals of the Wistar male adult, average weight of 175g, placed into four groups with five animals each group.
Trichotomy, antisepsis and skin lesion were carried in all animals using a disposable microtome blade, previously anesthetized.
The first group (saline control group) and second group (sodium diclofenac control group) received no iontophoresis and third
group (saline experimental group) and fourth group (experimental group sodium diclofenac) received the application of
iontophoresis. All groups received 6 therapy applications, being removed the skin of each animal and evaluated the resulting scar
tissue by the scoring system 0-3 with observation of the epithelium, the presence of hemato-fibrinous and collagen fibers through
the light microscopy by Mallorys stain. The control group animals showed a great lack of epithelial lining in 40%, hematofibrinous present in 100% of the animals and the collagen fibers had 40% the formation of fibers, but in a disorderly fashion.
About 60% of sodium diclofenac experimental group showed a marked formation of the epithelium, with 100% showing the
absence of hemato-fibrinous and 100% with the formation of arranged and orderly collagen fibers.
Conclusions:
Based on our results, we conclude that the efficiency and effectiveness of the technique associated with drug actions promote the
remission of the healing of animals involved in this experiment.
Keywords: Electrotherapy, Iontophoresis, Healing, Sodium diclofenac
Financial Support: FIC, PIBIC-CNPq
Resumo:09-035
BRADYKININ B2 RECEPTOR INVOLVEMENT IN HUMAN RED BLOOD CELLS INVASION BY PLASMODIUM
FALCIPARUM
Souza-silva, L. 1; Ferreira-dasilva, C. T. 1; Caruso-neves, C. 1,2; Pinheiro,a. A. S. 1,3

1
Instituto de Biofsica Carlos Chagas Filho, IBCCF/UFRJ
2
Institutos Nacionais de Cincia e Tecnologia , INCT/INBEB/CNPq/MCT
3
Institutos Nacionais de Cincia e Tecnologia, INCT/INPeTAm/CNPqMCT
Objectives:
P. falciparum is a protozoan parasite that causes the most virulent form of human malaria. It infects both liver and red blood
cells, but the erythrocytic stage of infection is responsible for all symptoms and pathologies associated with the disease. We have
recently shown that components of the renin-angiotensin system (RAS), specifically Angiotensin II (Ang II), through its
conversion into Ang-(17), decreases infection of new erythrocytes during P. falciparum blood stage (PLos One; 6, e17174,
2011). The kallikrein-kinin system (KKS) is another complex pathway linked to RAS by a number of molecules including the
angiotensin converting enzyme (ACE). Studies showing a direct relationship between plasmatic level of bradykinin, an effector
molecule of KKS, and severe malaria were first demonstrated in 1966 (Ann. Trop. Med. Parasitol. 60; 304, 1966). More recently,
a new KKS inhibitor, designated anophensin, was identified in the salivary glands of the malaria vector mosquito, Anopheles
stephensi, indicating a possible role of KKS components in the life cycle of malaria parasite (Ins. Bioc. Mol. Biol. 35; 466, 2007).
In the present work, we demonstrate the role of bradykinin in the modulation of P. falciparum erythrocytic cycle.
Erythrocytic asexual stages of P. falciparum, W2 strain, were maintained in continuous culture in RPMI 1640 medium in the
presence of A-type human blood and 10% A-positive human serum, 50 gml gentamicin at 45% hematocrit. Cultures
synchronized at the schizont form were treated or not with bradykinin or other drugs and parasitemia was measured by counting
of the ring forms in blood smear, after 24h. Bradykinin (1014108M) reduced erythrocyte invasion by parasite in a dose
dependent manner with maximum effect of 51% inhibition observed at 108 M, when compared to control (n=8). This effect was
completely reversed by 108M HOE-140, an antagonist of B2 bradykinin receptor (n=5). The same phenomenon was observed
when cultures were treated with 107M A779, a specific Ang-(1-7) receptor antagonist (n=8). The presence of B2 receptor on the
surface of erythrocytes was evaluated by immunoblotting, using membrane fraction of normal cells and was detected at the
molecular weight of 50kDa. Captopril (106M), a well-known ACE inhibitor, responsible to increase the plasmatic level of
bradykinin, reduced invasion in 40% (n=5). The concomitant addition of increasing concentrations of Ang-(1-7) (1012-106M)
together with decreasing concentrations of bradykinin (108-1014M), showed an additive effect of both peptides, with inhibition
levels close to 50%, in relation to control (n=5).
Conclusions:
Taken together, our results suggest a cooperative role of bradykinin and Ang-(1-7) in the impairment of P. falciparum
erythrocytic cycle. The bradykinin effect depended on B2 and AT(1-7) receptors activation, indicating a possible
heterodimerization of both receptors in the erythrocyte membrane.
Keywords: Plasmodium falciparum, Red blood cells invasion, kallikrein-kinin system, Bradykinin B2 receptor
Financial Support: FAPERJ, CAPES, INBEB, INPETAm and CNPq
Resumo:10-031
EFFECTS OF N-ACETYLCYSTEINE AND/OR DEFEROXAMINE ON MYONECROSIS IN DYSTROPHINDEFICIENT MUSCLE FIBERS OF MDX MICE.
Moraes, L. H. R. ; Dias, P. ; Burgos, R. R. ; Neto, H. S. ; Marques, M. J. ; Minatel, E.
Anatomia, Biologia Celular, Fisiologia e Biofsica / IB, Unicamp
Objectives:
Duchenne muscular dystrophy (DMD) is caused by deficiency of the cytoskeletal protein dystrophin. The mdx mouse is an
animal model of DMD that also lacks dystrophin. In mdx mice and in DMD, oxidative stress appears to contribute substantially to
the skeletal muscle damage. The current study investigates the role of the antioxidant N-acetylcysteine (NAC) and the iron
chelator deferoxamine (DFX) or NAC-DFX in combination on myonecrosis in dystrophin-deficient muscle fibers of mdx mice.
Mdx mice (14 days old) received daily intraperitoneal injections of saline, NAC (150mgkg), DFX (150mgkg) or NAC plus DFX
for 14 days. C57BL10 mice were used as controls. Forelimb muscle strength was evaluated at the beginning and end of treatment
by grip strength meter. After the treatment, the sternomastoid (STN), diaphragm (DIA) and tibialis anterior (TA) muscles were
removed and used for the quantification of necrotic fibers (labeled with Evans blue dye)(EBD), regenerated muscle fibers with
central nuclei and inflammation area. Creatine kinase (CK) levels were analyzed for biochemical evaluation of muscle fiber
degeneration. There was a significant increase in strength in mdx mice treated with NAC plus DFX compared to saline-treated
mdx mice (2,560,53gg in NAC plus DFX treated mdx mice vs 2,030,46 gg in saline mdx mice, p0.02; Students t-test.). In
contrast, NAC or DFX treatments did not change forelimb muscle strength in mdx mice. NAC, DFX and NAC plus DFX
treatments caused a significant decrease in EBD staining in DIA, STN and TA muscles in mdx mice (p0.05; Students t-test)
compared to saline-treated mdx mice. In STN muscle, a significant decrease of fibers with central nuclei and inflammatory area
was observed after treatment with NAC, DFX and NAC plus DFX (p0.05; Students t-test). In TA muscle, NAC and NAC plus
DFX treatments significantly reduced the percentage of fibers with central nuclei in mdx mice compared to saline-treated mdx
mice (p0.05; Students t-test). Although was observed a reduction in inflammatory area of TA mdx muscle after all treatments,
the decrease was not statistically significant (p0.05; Students t-test). In DIA muscle, NAC treatment reduced significantly the
percentage of fibers with central nuclei in mdx mice compared to saline-treated mdx mice (p0.05; Students t-test). Furthermore,
the inflammatory area decreased in DIA muscle of mdx mice after treatments with NAC, DFX and NAC plus DFX (p0.05;
Students t-test).
Conclusions:
These results suggest that NAC, DFX and NAC plus DFX treatments protects STN, TA e DIA muscles against myonecrosis in
mdx mice.
Keywords: mdx mouse, oxidative stress, antioxidants
Financial Support: CAPESProex; FAPESP (#2010/010874; #2008/507313); FAEPEX.
Resumo:10-032
EVALUATION OF LIVER TISSUE IN DIABETIC RATS INDUCED BY STREPTOZOTOCIN AND TREATED WITH
AQUEOUS EXTRACT OF EQUISETUM PYRAMIDALE.
Santos, S. E. 1; Aquino, J. P. 1; Silva, M. B. D. 1; Dourado, D. M. 2; Pereira, D. M. 4; Silva, B. A. K. D. 4;

Coelho, R. M. 3; Bento, L. M. A. 2
1
LABORATRIO DE FISIOLOGIA EXPERIMENTAL, UAU
2
LABORATRIO DE TOXINOLOGIA E PLANTAS MEDICINAIS, UAU
3
LABORATRIO DE PRODUTOS NATURAIS, UAU
4
DOCENTE DO CURSO DE FISIOTERAPIA, UAU
Objectives:
This study has as aim to evaluate the effect of plant extract E. pyramidale em the liver of diabetic rats.
Wistar Mice with body weight of 250g in average, inducted to Diabetes Melitus(55 mg/kg STZ). They were divided into 4
groups: Extract control(EC,n=5), saline control(CSn=5), diabetc extract(DE,n=6) and diabetc saline(DS,N=5).By DE and CE
Gavage they received extract of E. pyramidale and CS and DS received saline, both at the dose of 1ml/for a day. After 40 days
the animals were euthanized and samples of the liver removed, fixed in 10% buffered formalin, processed, embrdded in paraffin
to be sectioned to 5mm and evaluated by histochemical staining reaction of schiff. By capturing 10 fields per individual
ramdomly, chooosing the field hepatocytes with cytoplasm filed with glycogen in the 40 x objective for statistical analysis. The
test used for comparison of glycogen in hepatocytes was between groups analysis of variance ANOVA with Tukey post-test(p
Conclusions:
In the measurement of glycogen in hepatocytes controls had lower numbeer compared to diabetcs, demostrating that glycogen
accumulates in the liver, preventing its proper functioning. There were no histological changes in the controls, but tissue necrosis
in diabetcs patients. Morphologically there is no significant difference between the groups. Futher studies are needed to evaluate
the liver tissue in diabetc rats treated with E. pyramidale.
Keywords: diabetic, plants, Equisetum pyramidale, streptozotocin, liver
Financial Support: pic aesa: universidade anhanguera uniderp
Resumo:10-033
EVALUATION OF HEMATOLOGIC AND BIOCHEMICAL PARAMETERS OF RATS SUBJECTED TO SECONDDEGREE THERMAL BURN
Pereira, D. S. T. 1; Lima-ribeiro, M. H. M. 2; Cunha, C. R. A. 1; Carneiro-leo, A. M. A. 2; Correia, M. T. S.

3
1
Programa de Ps-Graduao em Cincias Biolgicas, UFPE

Programa de Ps-Graduao em Biocincia Animal, UFRPE
3
Departamento de Bioqumica, Centro de Cincias Biolgicas., UFPE
2
Objectives:
Determine the biochemical and hematological parameters of rats subjected to second-degree thermal burn.
Seven rats (Rattus norvegicus), albino Wistar male were obtained from the Vivarium Creation of the Department of Nutrition,
Federal University of Pernambuco and the experiments conducted in the Animal Experimental Center of Experimental Surgery at
that institution. All procedures related to animal use were performed according to the standards recommended by the Brazilian
College of Animal Experimentation (COBEA) and approved by the Ethics Committee on Animal Experiments of UFPE
(Protocol: 23076.015015/2009-31). The animals were between 9 and 10 weeks old, weighing on average 268 to 283 g, were preanesthetized with atropine sulfate at a dose of 0.04 mg / kg intramuscularly. After 10 minutes we used an anesthetic combination
of ketamine 10 % (90 mg / kg) with xylazine 2 % (10 mg / kg) intramuscularly and underwent shaving of the back, by the pull of
the direct and antisepsis with polyvinylpyrrolidone-iodine 1 %. The second-degree thermal injury was produced with a solid
aluminum bar 1 cm in diameter. On the 7, 14, 21 and 28 days after surgery, the animals were anesthetized for blood collection
by cardiac puncture for hematological and biochemical, followed by euthanasia with sodium pentobarbital (100 mg / kg
intraperitoneally). During the study period the lesions showed no signs of infection. The bleeding, when this was related to
damage caused by the animal itself. The hematological values obtained in this study showed no significant changes as a function
of induction of the burn during the period analyzed (Erythrocytes: 7.6 0.48; Hemoglobin: 13.65 0.5; Platelets: 846400 0.71,
leukocytes: 7980 0.71; Basophils: 0.2 0.05; Eosinophils: 1.38 0.18; Lymphocytes: 82.37 0.83; Monocytes: 1.9 0.2),
revealing themselves but normal values in rats (Medicina veterinria, 3(2):1-8, 2009). The rats, like other mammals, have to
maintain strict control of the internal environment thus ensuring the homeostasis (COBEA, 2003). It is known that rats can
produce changes in these parameters as a result of pathological processes or external factors such as sex, ancestry, age, diet,
handling and environment (Ceres, 56(1):051-057, 2009). However, average values of biochemical parameters analyzed in this
study were consistent with previously reported data specific to normal animals (Calcium: 10.04 0.42; Pro-Thrombin: 9.94
0.16; Fibrinogen: 457.32 0.25; Alkaline Phosphatase: 212.68 0.52; Glutamic Oxalic Transaminase: 180.02 0.35; Glutamic
Pyruvic Transaminase: 53.28 0.41; Gamma-Glutamyl Transpeptidase: 5.76 0.23, Creatine: 0.54 0.04; urea: 46.34 0.04
and Amylase: 842.06 0.48).
Conclusions:
Based on the results obtained did not observe significant changes in hematological and biochemical parameters evaluated, not just
differences occurring due to thermal burn injury in the second degree.
Keywords: Biochemical parameters, Burn, Hematologic parameters, Rats
Financial Support: FACEPE
Resumo:10-034
ORGANIZATION OF COLLAGEN AND MT1-MMP EXPRESSION IN THE PERIODONTAL LIGAMENT OF RAT
INCISORS UNDER EXPERIMENTALLY ALTERED ERUPTION.
Omar, N. F. 1; Gomes, J. R. 2; Neves, J. S. 1; Narvaes, E. A. O. 1; Novaes, P. D. 1

1
Departamento de Morfologia, FOP/UNICAMP
2
Departamento de Biologia Estrutural, Molecular e Gentica , UEPG
Objectives:
In the rodent teeth have been demonstrated that the hypofunction condition increases the rate of tooth eruption. The constant
movement that occurs in rat incisor eruption is accompanied by intense activity in periodontal ligament remodeling. The ligament
extracellular matrix remodeling occurs by direct action of enzymes known as metalloproteinases (MMPs). In this sense, this
study investigated: the eruption rate of rat incisors; the periodontal ligament resistance strength in eruption moviment; the
collagen organization and the MT1-MMP expression in periodontal ligament.
Twenty male Wistar rats had their lower left incisors cut, every two days, at the interdental papilla level, using high-drill rotation,
after anesthesia by halothane, producing hypofunction condition (HP) during 7 days. Other twenty rats were kept in normal
condition of eruption were used as control (N). The eruption rate was measured with an ocular millimeter from the gingival
margin to the top of the tooth in group HP. In the N, the measure was made from the gingival margin to a mark made in the tooth,
previously. After 7 days, ten animals of each group were killed by cervical dislocation under anesthesia, and the tensile strength
of the periodontal ligament was measured using an algometry. After extracting, the periodontal ligament of the animals was
scraping with a periodontal curette, collected in microtubes containing Tris buffer and used to assess expression of MT1-MMP by
western blotting. It was applied 20g of samples total protein in poliacrylamide 12% gel. For polarization analyses 10 animals of
each group were killed by perfusion through the heart, the mandibles were removed included in paraffin and the periodontal
ligament of mesial, lingual and distal faces were analyzed in polarized light microscopy. In the hypofunctional group (HP) the
eruption rate (p
Conclusions:
In this study, we conclude that in hypofunction condition there is an increase in rate eruption with degradation of collagen in the
periodontal ligament added to an increase in MT1-MMP expression that leads to a lower resistance in this tissue.
Keywords: Rat, Periodontal ligament, Collagen, Metalloproteinases
Resumo:10-035
CHANGES IN THE COLLAGEN CONTAINING TISSUE FROM RATS CHRONICALLY TREATED WITH
SUPRAPHYSIOLOGICAL DOSE OF GLUCOCORTICOID
Munhoz, A. L. B. 1; Zavan, B. 1; Paffaro Jr, V. A. 1; Pimentel, E. R. 2; Vilela, A. S. B. 1

1
Biomedical Sciences Institut, UNIFAL
2
Institut of Biology, Unicamp
Objectives:
To investigate the changes related to non collagenous proteins, extracellular matrix proteoglycans and glycosaminoglycans ,
fibroblasts in the calcaneal tendon and cornea from male and female rats after prolonged treatment with supraphysiological doses
of glucocorticoid.
We have used 20 Wistar rats, divided into four groups of 5 animals each group, control males, treated males, control females and
treated females. The treated group received during five weeks intraperitoneal injection of betamethasone dipropionate +
betamethasone disodium phosphate (0,075 mg/kg). The control group animals received weekly equivalent volume of saline. After
the sixth week, animals were sacrificed by injection of 10% chloral hydrate (0.3 mL/100 g body weight) and their corneas and
Achilles tendons were collected. For morphological study, histological sections of 7m thickness were stained with hematoxylin
and eosin. These slides were used for counting the tendon fibroblasts, using the NIS-Elements software. The biochemical analysis
was performed extracting extracellular matrix components and these extracts were analyzed by SDS-PAGE as well as dosage of
non collagenous proteins and glycosaminoglycan (GAG) sulfate. The analysis of GAGs was undertaken on tissue with papain
digestion and subsequent agarose gel electrophoresis in propylenediamine buffer. Statistical analysis was performed by analysis
of variance (ANAVA) followed by the comparison test of Tukey. When the assumptions of normality and homoscedasticity did
not met it was applied the nonparametric Mann-Whitney-Wilcoxon test. The treatment increased the amount of tendon non
collagenous proteins in female, but did not change it in males. The cornea has the amount of non collagenous proteins decreased
significantly in both groups. The quantification of tendon fibroblasts nuclei has not revealed significant differences between
experimental groups. In these experimental conditions, both males and females had not altered the amount of glycosaminoglycans
present in tendon and cornea. However, the results suggest that there are less of these compounds in the cornea of females
compared with males. In tendon, the GAG found in control and treated groups was dermatan sulfate. The treatment seems to
change the type of GAG in cornea, from keratan sulfate to dermatan sulfate.
Conclusions:
The glucocorticoid treatment appears to increase the amount of tendon non collagenous proteins in females, however, it does not
change in males, whereas in cornea of treated groups a significant decrease in the amount of non collagenous proteins are
presented in extracellular matrix. In both males and females, the treatment did not seem to change the amount of
glycosaminoglycans present in tendon and cornea. However, the obtained amount of glycosaminoglycans in cornea suggests that
females have less of these compounds compared to males.
Keywords: glucocorticoid, extracellular matrix, tendon, cornea, rats
Resumo:10-036
DIFFERENT RESPONSES OF INTRAMEMBRANOUS AND ENDOCHONDRAL AUTOGENOUS ONLAY BONE
GRAFTS TO LOW LEVEL LASER THERAPY
Biguetti, C. C. 1; Caviquioli, G. 1; Holgado, L. D. A. 1; Marquardt Filho, E. J. 1; Moreschi, E. 1; Ribeiro

Junior, P. D. 1; Duarte, M. A. H. 2; Matsumoto, M. A. 1
1
Depto. Cincias da Sade/ Universidade Sagrado Corao, USC
2
Faculdade de Odontologia de Bauru/Universidade de So Paulo, FOB/USP
Objectives:
Some doubts are still raised upon the behavior of different types of autogenous bone used for craniomaxillofacial reconstruction
in relation to their embryologic origin. The aim of this study was to evaluate the process of incorporation of intramembranous
(IM) and endochondral (EC) bone grafts, associated or not to low level laser therapy (LLLT).
Thirty-two male New Zealand rabbits underwent onlay autogenous bone grafts retrieved from the calvaria and the iliac crest and
were divided in four groups: Control groups Calvaria (CC) and Iliac (CI) negative controls, and Experimental groups Calvaria
(ExC) and Iliac (ExI)- exposed to LLLT at 16 J/cm2. After 7, 14, 30, and 60 days the grafts retrieved for microscopic
morphological analysis of the interface, measurement of the grafts thickness, and histochemical analysis for the detection of
osteoclasts in the surface of the grafts. Bone grafts incorporation occurred uneventfully in both IM and EC grafts. Differences in
the resorption process were evident in iliac graft, presenting resorption level of 40% in CI against 8% in ExI at the 14th day.
After 30 days, the resorption level was maintained at 41%, rising to 70% at the 60th day period in CI, while ExI presented 15%
and 45% of resorption after 30 and 60 days, respectively. No significant differences were noted in the calvaria graft. A significant
higher number of osteoclasts was observed after 7 days.
Conclusions:
It was concluded that LTB exerts a more efficient biomodulative activity on EC graft, preventing the resorption process in an
important level.
Keywords: Low level laser therapy, Rabbits, Bone grafts, Intramembranous, Endochondral
Financial Support: FAPESP 2008/11485-7; 2009/14989-9
Resumo:10-037
THE REPAIR OF RABBITS CALVARIAL BONE DEFECTS USING BIOACTIVE VITROCERAMIC
BIOSILICATO
Caviquioli, G. 1; Biguetti, C. C. 1; Holgado, L. D. A. 1; Saraiva, P. P. 1; Kawakami, R. Y. 1; Renno, A. C. M.

1
; Matsumoto, M. A. 1
1
Universiade Sagrado Corao, USC
2
Universidade Federal de So Paulo, Campus Baixada Santista, UNIFESP
Objectives:
A number of biomaterials have been developed in order to act as bone substitutes. In the present study, the behavior of a new
granulate bioactive vitroceramic in bone repair defects was evaluated. Thirty rabbits underwent surgical confections of defects in
parietal bones, distributed into 4 groups, according to the filling material: 1 blood clot (Control), 2 particulate autogenous
bone, 3 bioactive vitroceramic (Biosilicato), and 4 - bioactive vitroceramic and particulate autogenous bone.
Specimens were retrieved after 7, 14, and 30 days for microscopic morphological analysis. Similar patterns of new bone
formation were observed amongst experimental groups, with a more evident woven bone deposition at day 14, directly upon the
non-viable bone grafts particles, as well as on the biomaterial granules, and bone maturation at day 30. Despite the foreign body
reaction induced in the presence of the vitroceramic, evident from the 14th day, no deficit on bone formation and maturation
occurred during the process. The presence of both biomaterial and bone graft offered a greater volume in the region of the defect
until the last period, differently from the Control Group where small trabeculas surrounded by connective tissue could be seen,
resulting in a repaired defect thinner than the original bone.
Conclusions:
Biosilicato presented a satisfactory behavior during the repair of bone defects, permitting new bone formation directly upon its
granules surfaces when used associated with bone graft or alone.
Keywords: Bone repair, Rabbit, Bioactive glass, Biomaterial
Financial Support: FAPESP 2009/17294-1
Resumo:10-038
BONE REPAIR INVESTIGATION IN DEFECTS GRAFTED WITH HYDROXYAPATITE COMBINED WITH
LASER THERAPY IN RATS SUBMITTED TO PASSIVE TOBACCO EXPOSURE
Franco, G. R. ; Figueira, L. A. ; Ribeiro, D. C. P. ; Miguel, N. M. ; Cunha, M. R.

Depto Morfologia e Patologia/Faculdade de Medicina de Jundia, FMJ
Objectives:
Defects with bone mass loss are frequently treated with bone autografts. Endografts of bones using biomaterials, such as
hydroxyapatite also have been used for the same purpose, replacing autografts. In addition to these biomaterials that represent an
excellent alternative and coadjuvant in bone reconstruction, laser therapy has been shown to contribute to accelerate the fracture
repair by improving local microcirculation and increasing collagen synthesis However, bone tissue health conditions are basic for
osteointegration of the implant. Thus, excessive tobacco consumption, either as an active or as a passive smoker, may harm the
process of regeneration due to its deleterious effects to bone tissue. The objective of this study was to evaluate the process of
bone regeneration stimuled with laser therapy when hydroxyapatite granules are implanted in bone defects of the femur of rats
submitted to passive tobacco exposure.
Porous hydroxyapatite granules were implanted in bone defects produced in the left distal femoral epiphysis of twenty rats Wistar
(Rattus norvegicus) subjected to prolonged passive tobacco exposure during eight months. Immediately after implantation of the
biomaterials, gallium-arsenide laser irradiation was applied to the recipient area at an intensity of 5.0 J/cm2. The rats were
divided into four groups: animals control receiving hydroxyapatite implants without (group G1) and with (group G2) laser
therapy, animals treated (passive tobacco) receiving hydroxyapatite implants without (group G3) and with (group G4) laser
therapy. After eight weeks of the bone implant with the biomaterial, the animals were sacrificed and the specimens of the implant
region were submitted to routine histological testing, and maintained in paraffin blocks for histological analysis. In the results,
was observed good radiopacity of the implant recipient area and of the hydroxyapatite granules in all of studied groups.
Formation of new bone around the hydroxyapatite granules showing characteristics of trabecular and cortical bone was observed
in the studied groups controls (G1 and G2). In the groups submitted to passive tobacco exposure (G3 and G4), the hydroxyapatite
granules were involved by connective tissue without bone neoformation.
Conclusions:
The passive tobacco exposure injures the bone neoformation in skeletal defect and the laser therapy protocol used was not
adequate to stimulated the osteogenic process.
Keywords: HYDROXYAPATITE , LASER, TOBACCO, BONE, REGENERATION
Financial Support: FAPESP/ 2010/ 05769-2
Resumo:10-039
QUANTIFICATION OF OSTEOCLASTS INTO DENTAL ALVEOLI OF WISTAR RATS TREATED WITH
EXTRACTS OF PROPOLIS PURE WITH AND WITHOUT CONTAMINATION BACTERIAL
LIPOPOLYSACCHARIDE
Pereira, Y. C. L. 1; Iyomasa, M. M. 2; Issa, J. P. M. 2; Ervolino, E. 4; Mishima, F. 2

1
Faculdade de Odontologia de Piracicaba, FOP UNICAMP
2
Faculdade de Odontologia de Ribeiro Preto, FORP USP
3
Faculdade de Ciencias Farmaceuticas de Ribeirao Preto, FCFRP
4
Faculdade de Odontologia de Araatuba, FOAR
Faculdade de Medicina de Ribeiro Preto, FMRP

Faculdade de Odontologia de Ribeiro Preto , FORP USP
Objectives:
Aim: Propolis is a resinous substance, which antibacterial, antiinflammatory, antiviral, antifungal, immunostimulating healing
and local anesthetic has been considered for clinical use. The lipopolysaccharide (LPS) is recognized as an endotoxin and may
induce inflammatory processes. Our objectives were to analyze, in vivo, the action of Pure Propolis Extract (EPP) in dental
alveoli or not contaminated with lipopolysaccharide (LPS) bacteria.
Methods and Results: For in vivo study, 14 rats were submitted to extractions of the superior first molars, right and left, which
immediately had the right dental socket contaminated with 0.1 mL of lipopolysaccharide (LPS) (100g/kg) and maintained
without such contamination. The groups (n=7) for each treatment after 2 weeks were analyzed: GI-Negative Control (NC) - no
treatment, GII-Treated with Pure Propolis Extract (EPP). The bone samples were removed, demineralized, processed by routine
histologic technique, submitted to systematic sections (6 m of thick) for TRAP immunohistochemical staining. The number of
osteoclastic cells stained were counted. The relationship between bone formation and the number of clastic cells was obtained
and submitted to statistical analysis. Observing the normality of data distribution, we proceeded to the factorial ANOVA and
Tukey-Kramer test (p
Conclusions:
Conclusion: We conclude that at 14 days of treatment, it was observed statistical difference between groups with green propolis
contaminated with LPS.
Keywords: TRAP, Bone, osteoclasts, lipopolysaccharide, wistar
Resumo:10-040
INCIDENCE OF DENTAL CALCULUS (DC) AND PERIODONTAL DISEASE (PD) IN BEAGLE DOGS FROM THE
CENTRAL ANIMAL FACILITY (BIC) OF THE FEDERAL UNIVERSITY OF SANTA CATARINA (UFSC), BRAZIL.
Lorenzo, M. A. 1; Santos, A. C. 1; Bello, L. F. C. O. 2; Santos, G. A. 1; Rothstein, J. M. J. 1

1
Biotrio Central/Universidade Federal de Santa Catarina, UFSC
2
Mdico Veterinrio Autnomo, VetBello
Objectives:
Periodontal disease affects 85% of adult dogs (Clin. Vet. 1; .6-7, 1996), one of the most common infirmities in small animal
clinics. It results of the inflammatory response to dental plaque, which is the accumulation by bacteria on the tooth surface, which
is subsequently mineralized, forming the DC (tartar) (Odont. Peq. An., 2010). This disease can generate clinical signs such as bad
breath, drooling, tooth mobility, and unusual signs like dysphagia, anorexia, fractures and osteomyelitis (Comp. Cont. Educ.
Pract. Vet. 12; 951-960, 1990). There is evidence that this illness can also determine the involvement of vital organs (heart, liver,
kidneys) and joints (Clin. Vet. 1; .6-7, 1996). This study determined the incidence of DC and PD for dental and age group in
beagle dogs reared under controlled management and feeding at the BIC/UFSC.
A total of 25 beagle dogs of the breeding herd kennel BIC/UFSC, 19 females and 6 males with a mean weight of 17 kg and age
1.5-9 years. This animals were anesthetized with tiletamine/zolazepam 5% - 10 mg/kg, xylazine hydrochloride 2% - 0.5 mg/kg
and atropine sulfate 0.05% - 0.04 mg/kg, IM and underwent a clinical examination of oral cavity for measuring the degree of DC
and PD through the table of scores LASCALA and MOUSSALLI (Period. cln.,1980) and BEARD & BEARD (Vet. Clin. of
North Amrica 19, 1,1989), respectively. Were observed that the incisors (I) were the least affected tooth by the DC (score
0:14,171,76/n=300) and the molars (M) had a higher incidence of score 1(12,91,18/n=250); already, canines (C) and premolars
(PM) were severely affected with prevalence of scores 2 (13,50,95/n=100) and 1 (13,941,25/n=400), respectively. In the
presentation of PD, the results were as follows: I predominated the score 0 (17,921,04/n=300), C and PM score 1
(1,250,75/n=100;7,310,86/n=400, respectively) and M scores 0 and 1 (10,51,91 and 11,81,33/n=250,respectively). In the
evaluation of DC and PD, associated to the animal age, it was observed in both the trend rise in scores with increasing age. The
I have a significant increase in the score 1(DC:5,160,51;PD:3,750,39/n=132), at the age of 3-6 years. The M showed a
decrease in score 0 (DC: 0,30,15; PD: 1,20,32/n=60) in the group with of 6-9 years old, compared with other groups, although
a predominance of score 1 for DC (5.7 0.74 / n = 110) is observed in the group with 3-6 years. The C and PM have the
dominance of the score 1 for DC already at age 0-3 years (5.0 0.57 / n = 32, 5.1 0.56 / n = 128, respectively), with C
exhibiting the same trend for PD (6,750,25/n=32), but not the PM, where the PD is expressed significantly at age 3-6 years
(5,750,33/n=176).
Conclusions:
According to the results, it appears that the dental groups most affected by DC and PD are the C and PM. These changes begin
early in life of dogs (0-3 years), and considering the consequences on the health care of them, justify the adoption of preventive
measures to maintain health and welfare of these animals.
Keywords: periodontal disease, dental calculus, dogs
Resumo:12-029
THE INCREASED LEVELS OF PHOSPHO-EIF2-ALPHA INDUCES PHOTORECEPTOR DEGENERATION IN
MURINE MODELS OF RETINITIS PIGMENTOSA(RD1)
Gonalves, B. S. ; Munk, N. ; Vieira-barroso, L. ; Chiarini, L. B.

Objectives:
The retinitis pigmentosa is characterized by death of photoreceptors in the retina what causes blindness. Therefore, the analysis of
the mechanisms responsible for the death of retinal photoreceptors is critical for the development of therapeutic strategies.
Recently, endoplasmic reticulum stress, activation of PERK and an increase of phosphorylation of eIF2alpha were described in
rd1 mice, a mutant model of retinitis pigmentosa. However, the role of PERK/eIF2alpha pathway, in this context, have not been
evaluated. Our purpose was to test the role of phosphorylation of eukaryotic translation initiation factor 2 alpha subunit
(eIF2alpha) in the degeneration of photoreceptors in retinal tissue from mice RD1 C3H-HeJ, a murine model of retinitis
pigmentosa. This is an animal homozygous for mutation in the gene encoding the subunit of cGMP phosphodiesterase. This is
one of the most common mutations associated with autosomal recessive retinitis pigmentosa in humans.
The degeneration of photoreceptors was assessed by measuring the thickness of the Outer Nuclear layer (ONL, formed by the
photoreceptors) and by detection of DNA fragmentation in situ (TUNEL assay). C57-BL6 wild-type mice were used as control
group. Retinal explants were maintained in vitro for 24 hours in the presence of salubrinal, an inhibitor of the dephosphorylation
of eIF2alpha. In our experimental conditions, the degeneration of the photoreceptor layer of RD1 C3H-HeJ begins on the
fifteenth day post-natal (P15) and is time dependent. We found by western blot an increase of phosphorylated eIF2alpha content
that coincides with the period of the degeneration of the photoreceptor cells (P15). The treatment of retinal explants from
rd1C3H/HeJ mice with salubrinal, the inhibitor of the dephosphorylation of eIF2-alpha, increases the incidence of TUNEL
positive photoreceptors and decreases the thickness of ONL.
Conclusions:
These results indicate that the accumulation of phosphorylated eIF2alpha induces the death of photoreceptors in this model of
retinitis pigmentosa.
Keywords: Retina, Apoptosis, UPR, eIF2, Retinitis Pigmentosa
Financial Support: CNPq, FAPEJ, UFRJ-PIBIC.
Resumo:12-030
INVOLVEMENT OF THE P53 TUMOR SUPPRESSOR PROTEIN IN RESVERATROL-INDUCED APOPTOSIS IN
MCF-7 AND H1299 CELLS
Costa, D. C. F. 1; Malheiros, M. S. 1; Campos, N. P. C. 1; Casanova, F. A. 1; Sanches, D. 1; Fialho, E. 2;

Silva, J. L. D. 1
1
Instituto de Bioqumica Mdica - UFRJ, IBqM - UFRJ
2
Instituto de Nutrio Josu de Castro - UFRJ, INJC - UFRJ
Objectives:
Resveratrol (RV) is a promising chemopreventive agent, being able to modulate many cellular targets involved in cancer
signaling pathways. Since p53 has been suggested to be important for anticancer properties of RV, we investigated the RVinduced cytotoxicity in H1299 non-small lung cancer cells, which have a partial deletion of p53 protein-encoding gene, and
compared with MCF-7 breast cancer cells, which constitutively express wild-type p53.
RV (0-500M) reduced both H1299 and MCF-7 cells viability in a dose- and time-dependent manner. However, MCF-7 cells
were more sensitive than H1299, when exposed to RV at concentrations higher than 100M for 24h. RV (0-200M) also
increased p53 protein levels in MCF-7, measured by western blotting, without altering p53 mRNA, detected by RT-PCR, thus
suggesting a possible modulation of this protein by post-translational processes. Under these experimental conditions, RV
stimulated phospho-Chk2 and p21 in MCF-7. Moreover, in these cells, RV-induced cytotoxicity was partially mediated by p53
and involved caspases 9 and 7 activation, as well as PARP cleavage, which is indicative of apoptosis. In H1299, cytotoxicity of
RV was less pronounced, as determined by MTT and live/dead assays and, unlike MCF-7, cell death were not accompanied by
caspases activation. These results also corroborate the finding that MCF-7 cells were positively labeled by TUNEL after
exposition to 100M RV, but not H1299 cells at the same conditions. However, transient transfection of wild-type p53-GFP
renders H1299 more sensitive to the proapoptotic effects of RV.
Conclusions:
Modulation of p53 by RV seems to be an important mechanism by which this bioactive compound exerts its chemopreventive
effects. Our results suggest that H1299 requires p53 expression to undergo apoptosis in response to RV, similar to MCF-7 cells.
Keywords: resveratrol, cancer, MCF-7, H1299, p53
Financial Support: INBEB, FAPERJ, FAF and CNPq.
Resumo:12-031
INVESTIGATION OF YELLOW FEVER VIRUS-INDUCED ENDOPLASMIC RETICULUM STRESS
Sanches, D. 1; Campos, S. P. C. 1; Rocha, C. M. 1; Gonalves, B. S. 2; Chiarini, L. B. 2; Gaspar, L. P. 3;

Freire, M. S. 3; Silva, J. L. 1; Gomes, A. M. O. 1; Oliveira, A. C. 1
1
Instituto de Bioqumica Mdica, IBqM-CCS-UFRJ
2
Instituto de Biofsica Carlos Chagas Filho, IBCCF-UFRJ
3
Instituto de Tecnologia em Imunobiolgicos, ITI-FIOCRUZ-RJ
Objectives:
Flaviviruses are arboviruses and cause diseases like Dengue and Yellow fever. These viruses have a particular importance for
public health mainly in South America, Central America and Asiatic southeast. Virus-induced apoptosis is related to a
cytopathological consequence of an infection in vivo or in vitro. During apoptosis, some cellular mechanisms occur, DNA
fragmentation and release of messengers of the apoptotic pathways. The endoplasmic reticulum stress (ERS) can be triggered by
an accumulation of unfolded protein leading to stress response (UPR) with BiP-PERK complex dissociation. Once PERK is
dissociated from BiP it can lead to eIF2 phosphorilation, inducing CHOP overexpression. CHOP is a nuclear factor that leads to
expression and translocation of pro-apoptotic Bcl-2 proteins from the cytosol to the mitochondria. Yellow Fever Virus (YFV) and
other members of the Flaviviridae family are endoplasmic reticulum (ER)-tropic viruses that are dependent on the host ER to
translate, replicate and package their genome. Here, we investigate the endoplasmic reticulum stress induced by YFV.
We infected Vero cells with YFV using a MOI = 1 and analyzed the cell viability using LIVE/DEAD kit assay and lactate
dehidrogenase (LDH) activity assay. The expression of BiP, PhosphoeIF2 and CHOP was followed by Western-blotting in a
time-dependent manner after 24, 48 and 72 hours post infection. The YFV-induced ERS was confirmed by PhosphoeIF2
overexpression 24 hours post infection. We also observed a reduction of cytosolic BiP levels after 24 hours of infection,
suggesting ERS activation. CHOP overexpression was observed after 48 hours post infection. We analyzed the role of PERK
pathway activation during YFV-induced apoptosis by using PERK pathway inhibitor 4 phenylbutyric acid (4-PBA). We observed
that even using 4-PBA inhibitor, YFV still induces apoptosis after 96 hours post infection.
Conclusions:
Our results suggest that ERS could lead to apoptosis pathway activation through CHOP overexpression, but PERK activation
does not seem to be necessary to YFV-induced apoptosis.
Keywords: Yellow Fever Virus, Apoptosis, Endoplasmic Reticulum Stress, PERK
Financial Support: CNPq, CAPES, FAPERJ, FINEP/CT-INFRA, INBEB, PRONEX
Resumo:12-032
STUDY OF LOPAP (LONOMIA OBLIQUA PROTHROMBIN ACTIVATOR PROTEASE) AND DERIVED PEPTIDE
ACTIVITIES IN THE MODULATION OF HUVEC SURVIVAL.
Roedel, K. Q. ; Carrijo-carvalho, L. C. ; Flores, M. P. A. ; Chudzinski-tavassi, A. M.

Laboratrio de Bioqumica e Biofsica/Instituto Butantan, IB
Objectives:
Obtention of recombinant Lopap (rLopap), the most abundant protein found in the bristles of Lonomia obliqua caterpillars, and
study of cytoprotective effects of this protein and derived peptide (P4) in cultures of HUVECs (Human Umbilical Vein
Endothelial Cells). Lopap was characterized as a prothrombin activator serine protease which belongs to lipocalin family. The
members of this family present a wide functional diversity, among then, the modulation of cell growth, proliferation and survival,
being Lopap considered the only member of this family to possess proteolytic activity.
The recombinant protein was expressed with a histidine tag using the expression vector pAE, which was transformed on E. coli
BL21-DE3 by heat shock. The expression was done in 2xYT medium and induced with IPTG (0.5mM) for 3h at 37C. After that,
the recombinant protein was purified in Ni-Sepharose column and eluted with imidazole buffer (150mM), followed by desalting
in G-25 column to remove the imidazole salt. Steps of expression and purification were monitored by SDS-PAGE and the
concentration of rLopap was quantified by Bradford assay. The synthetic peptide (P4) was obtained by chemical synthesis in
solid phase according to a conserved domain of lipocalin family found in the Lopap sequence. The HUVECs were cultivated
following the method described by Jaffe et al. (1973), kept on deprived fetal bovine serum medium (RPMI + 1% FBS) and
treated with rLopap (10gml, 5gml and 2.5gml) and P4 (0.9gml, 0.3gml, 0.1gml, 0.03gml) (n=4) for 48 and 72h.
Cultures kept with 10% and 1% of serum and no treatment were used as control, being the cell viability measured by MTT assay.
The Lopap was obtained in the recombinant form with yield of 2mgL. When tested on HUVECs for 48h and compared with the
control 1% FBS, rLopap promoted an increase of 16% (p=0.0001) and 22% (p=0.002) in cell viability, at concentrations of
5gml and 10gml, respectively. On the other hand, P4 (0.1gml) increased cell viability by 11% (p=0.006). After 72h,
compared with the control 1% FBS, rLopap induced a viability increase of 11% at 2.5gml (p=0.001), 15% (p=0.0005) at 5gml
and 42% (p=0.0005) at 10gml. Cells treated with 0.1gml of P4 showed an increase in viability of 16% (p=0.01).
Conclusions:
Our results indicate that rLopap, as well as P4, have potential properties on modulation of cell survival, which is independent of
the proteolytic activity such as predicted for this molecule.
Keywords: Lopap, Peptide, HUVEC, Cytoprotection
Financial Support: FAPESP (201003516-0, 201000600-0)
Resumo:12-033
PROTECTIVE EFFECT OF GLUTAMINE AND ALANIL-GLUTAMINE ON CLOSTRIDIUM DIFFICILE TOXIN AINDUCED INJURY IN HUMAN AND RAT INTESTINAL EPITHELIAL CELLS IN VITRO
Santos, A. A. Q. A. 1; Neto, M. B. B. 1; Freire, R. 1; Santiago, T. M. 2; Ribeiro, R. 1; Brito, G. A. C. 3

1
Farmacologia, UFC
2
fsica , UFC
3
morfologia, UFC
Objectives:
Aim.: Clostridium difficille is one of the most frequent causes of diarrhea associated to the use of antibiotics. The increase of CDI
incidence and severity has been related to the upbringing of a high virulent strain NAP1/B1/027 which produces wider quantities
of toxin A (TcdA) and Toxin B (TcdB), as well as Binary Toxin (CDT). TcdA causes the Rho proteins family
monoglycosylation, which results in an actin cytoskeleton disorganization, cell rounding and loss of cellular adhesion. Glutamine
(Gln) is a critical requirement for the dynamic proliferating intestinal cell population but Gln has tendency to hydrolyze to
potentially toxic glutamate. Differently of the Alanyl-glutamine (AG) that is stable, well tolerated and repair intestinal injury in
vitro. We evaluated the effect of Gln or AG on the epithelial cell lines:HTC- 8 and IEC-6 following exposure to TcdA of
Clostridium difficile.
Four well slide chambers were seeded with HTC-8 cells in RPMI medium. After 48h, cells were incubated in media without Gln,
standard media or for 24 h with TcdA (10 ng / mL), TcdA + Gln (10 mM) and TcdA + AG (10 mM). After 24h of exposure, the
cells were fixed in 4.0% paraformaldehyde, and permeabilized with 0.1% Triton X-100. Cells were stained with FITC-phalloidin
and DAPI to performed confocal microscopy. Cell proliferation was evaluated using the tetrazolium salt WST-1 (4-[3-(4iodophenyl)-2H-5-tetrazolio]-1-3-benzene disulfonate). HCT8-cells were seeded and allowed to attach for 48 hours. Afterwards,
wells were incubated for 48h with 10ng/mL TcdA, TcdA + Gln (10mM) and TcdA + AG (10mM). Wells were incubated for 4
hours with tetrazolium salt and the absorbance was measured using an ELISA (450nm). In another set of experiment, cell culture
plates with 13 mm diameter glass coverslips. It was seeded with IEC-6 cells and grown for 24 h in DMEM media. Afterwards,
the wells were incubated for 24h with TcdA (10ng/mL) in DMEM without Gln or DMEM with Gln or AG. Cells were than fixed
in 4% formaldehyde for 14h. Slides were covered with a 15 mm gold film for conductivity by sputter and examined in a field
emission SEM. TcdA caused cell disruption, changes in F-actin cortex distribution, cell orientation and fragmentation of
cytoskeleton. Supplementation with both Gln and AG partially reverted the changes. Treatment with TcdA at 10ng/mL caused a
reduction of 8.4% on cell proliferation after 48h (p=0.01). Supplementation with 10mM of Gln and with of AG significantly
increased cell proliferation (12.7% and 13,2%, respectively) after 48h, compared to TcdA group; P<0.05.
Conclusions:
Gln and AG partially reverted these changes. Through SEM was observed that cells treated with TcdA shows a cellular
elongation and cytoskeleton disorganization. Those morphological alterations might be caused by disaggregation of actin
cytoskeleton as shown by confocal microscope. These results reinforce the Gln and AG potential as adjuvant therapeutic
measures in C.difficile enteritis.
Keywords: clostridium difficile, glutamine, Toxin A, alanil-glutamine
Financial Support: CNPq/PIBIC
Resumo:12-034
P2X7 RECEPTOR ACTIVATION INHIBITS THE PROLIFERATION OF CHICK RETINAL GLIAL PROGENITORS
IN CULTURE.
Silva, T. M. ; Ventura, A. L. M.
Neurobiology Department / Federal Fluminense University, UFF
Objectives:
ATP induces the proliferation of retinal glial progenitors at early periods of development through the activation of P2Y1
receptors (Intl. J. Devl. Neurosci., 20:21-27, 2002). However, in cultured astrocytes, ATP is also capable of inhibiting cell
proliferation via activation of P2X7 receptors (J. Neuroci. Res. 86:3096, 2008). In this study, we decided to evaluate whether
activation of P2X7 receptors is also able to inhibit ATP-induced proliferation of retinal progenitors in culture.
Cultures obtained from retinas of 7-day-old chick embryos were used in immunocytochemistry experiments and incorporation of
[3H]-thymidine assays. When retinal cells cultured for 2 days (E7C2) were treated for 24 h with 500 M ADP, an increase of
162% above the basal incorporation of [3H]-thymidine was observed. Addition of increasing concentrations of Bz-ATP to the
cultures, a selective agonist for the P2X7 receptor, was able to completely inhibit the proliferative effect of ADP at this stage of
development (in cpm/culture: basal = 905.1 124.8; ADP = 2373 159; ADP + 10 M Bz-ATP = 1456 154.7; ADP + 30 M
Bz-ATP = 1118 190.9; ADP + 100 M Bz-ATP = 692 89.6 p
Conclusions:
Although the two antagonists did not block the effect ob Bz-ATP, our data suggest that activation of P2X7 receptors by ATP
inhibits the proliferation of glial progenitors at an early stage of culture development.
Keywords: ATP, Proliferation, P2X7 Receptors , Glial Progenitors, Retina
Financial Support: FAPERJ, CNPq, Proopi-UFF, MCT-Pronex
Resumo:12-035
FERROMAGNETIC NANOPARTICLES: A TOOL TO LABELING AND TRACKING NEURAL STEM CELLS "IN
VIVO"
Rangel, B. 1,2,4,3; Santos, G. M. 1,2,4,3; Carvalho, A. B. 1,2,4,5; Rachid, R. 1,2,4,3; Attias, M. 1,2,5; Azevedopereira, R. L. 1,2,4,3; Mendez-otero, R. 1,2,4,3
1
Instituto de Biofsica Carlos Chagas Filho, IBCCF
2
Inst Nac de Cincia e Tecnol de Bio Estrut e Bioimagem, INBEB
4
3
Programa de Terapia Celular e Bioengenharia, PTCB
Objectives:
Neural stem cells (NSCs) are a source of new neural cells with capacity of proliferation and differentiation into neurons and glia.
This capacity opens a possibility for replacement cell therapy in neurological disorders. Preclinical and clinical research will
benefit from reliable in vivo tracking of transplanted cells. Here, we investigated the potency of superparamagnetic iron oxide
particles (SPION) to label NSCs, the effect of SPION on: 1) survival; 2) proliferation; 3) cell migration and 4) differentiation into
neural cells
NSCs were isolated from E12.5 Swiss mice and cultured as neurospheres in DMEM/F12 supplemented with B27 and epithelial
growth factor (EGF; 20ng/mL) and bFGF (10ng/mL). To label NSC, neurospheres were incubated overnight with 50ug/mL of
SPION (Endorem) and 0.1UI/mL of protamine sulfate. After 24 hours, SPION were observed manly in periphery of the
neurospheres. The areas of neuropheres with and without SPION were analyzed after 48 hours. We did not observe difference
between the areas of neurospheres indicating that SPION did not impair the growth of neurospheres. After 24 hours of labeling,
the neurospheres were dissociated and analyzed by FACS. Two different cell populations were found in the neurospheres by
setting forward and side scatters to linear amplification: 26-30% of the cells were high in forward and side scatter, while 10.412.5% of the cells were low in forward and side scatter. Twenty six percent of the high in forward and side scatter cells were
labeled with SPION, while 20.4% of the low in forward and side scatter cells were labeled. We observed that 44% of the cells
were stained by propidium iodide indicating high mortality after neurospheres dissociation. To access if the neurospheres labeled
with SPION could differentiate in neural cells, we cultured them onto coverslips previously coated with poly-L-lysine (10ug/mL)
and laminin (20ug/mL). To differentiate preferentially into neurons, the cells were cultivated in Neurobasal medium with B27
and N2 supplement. SPION are detected at 3 and 7 in neurons, astrocytes and oligodendrocytes from NSCs. In addition,
transmission electron microscopy (TEM) was performed in neurospheres 24 hours after incubation and SPION were detected
scattered in cellular cytoplasm.
Conclusions:
Our data indicate that NSCs can be labeled with SPION without impairment of neurospheres growth, differentiation and cellular
migration.
Keywords: Nanopaticles, Neural Stem cells, Neuron, Stem Cell
Financial Support: Edital CT-Biotecnologia/MCT/CNPq/MS/SCTIE/DECIT, CAPES, FAPERJ e INCT.
Resumo:12-036
EFFECT OF TREATMENT WITH PURIFIED BIOACTIVE FRACTION OF UNCARIA TOMENTOSA (WILLD) DC.
ON CELL GROWTH OF T24 HUMAN BLADDER CANCER CELL LINE.
Dietrich, F. ; Rockenbach, L. ; Cunha, F. M. D. ; Cappellari, A. R. ; Battastini, A. M. O.

Dep. Bioqumica- Universidade Federal do Rio Grande do Sul, UFRGS
Objectives:
Bladder cancer is the most common malignancy of the genitourinary tract and the effectiveness of treatments, most often, is
inadequate. Extensive studies in recent years, has demonstrated strong evidence for the involvement of purinergic signaling in the
physiology and in the tumor progression of the urinary bladder. Plant species of Uncaria tomentosa, widely used in traditional
medicine because of the many pharmacological properties attributed to it, already showed the presence of metabolites with
potential antitumor effect on different cell lines. Considering these informations, the objective of this study is to investigate the
antitumor effect of purified bioactive fractions of U. tomentosa on the ecto-nucleotidases activity and in cell proliferation on the
T24 bladder cancer cell line.

The alkaloidal fraction (AF) and triterpenoid fraction (TF) were obtained by maceration process from the barks of Uncaria
tomentosa. The T24 cell lines were cultured in RPMI culture medium supplemented with 10% fetal bovine serum in standard
conditions. For the evaluation of the antitumoral activity and its effect on cell proliferation, the cell number was determined after
treatments, where the cells were released, dissociated and counted by hemocytometer. The cell viability was evaluated by MTT
assay. The ecto-5'-nucleotidase/CD73 activity was determined by the inorganic phosphate released measured by Malachite Green
method, after different treatments of the bladder cells. Cell adhesion was also assessed, where adherent cells were fixed and
stained, and staining intensity was determined by optical density measurement. The anti-proliferative effect was found to TF after
treatment for 24, 48 e 72h at concentrations of 100 and 150g/mL, where the inhibition was 29,7% (9,49 SD) and 39,1% (3,98
SD); 58,5% (8,68 SD) and 68,5% ( 6,24 SD); 69,8% (4,33 SD) and 85,3% (5,59 SD), respectively. Still, in 72h significant
effect was observed at concentration of 50g/mL with inhibition of 37,2% (7,96 SD). The alkaloidal fraction did not affect cell
proliferation in any treatment. The treatment with TF significantly decreased cell viability, following the same profile observed
for cell proliferation. No change in the ecto-5'-nucleotidase/CD73 activity was observed when T24 cell lines were treated with
both fractions (TF and AF) for 10 min (direct effect) or when the cells were treated for 48h. In cell adhesion assay, there was an
increase of 40.8% ( 0.063 SD) in adhesion of T24 cells exposed to TF at concentration of 25g/mL, unlike the AF which was
not significantly changed on cell proliferation.
Conclusions:
Considering the proposed objectives, it becomes evident that this investigation showed promising biological properties for the TF
of Uncaria tomentosa, which a significant inhibition of T24 human bladder cancer cells proliferation when compared to AF.
However, the involvement of the enzyme ecto-5'-nucleotidase/CD73 was not observed after treatments with both fractions,
excluding its involvement in the death of T24 tumor cells. Furthermore, the increased adherence evidenced after treatment with
the TF, brings new perspectives for research, since the search for treatments that can improve adherence and, consequently,
inhibit the migration are important for decreasing the process of tumor metastasis.
Keywords: bladder cancer, Uncaria tomentosa, cell proliferation
Resumo:12-037
CREB SIGNALING IN NEURONAL SURVIVAL AND DIFFERENTIATION.
Santana, T. T. S. ; Arajo, J. A. D. M. ; Costa, M. R.

Departamento de Neurobiologia Celular, Instituto do Crebro, UFRN
Objectives:
In the last two decades, several approaches have been proposed to replace neurons lost in the course of central nervous system
diseases (Nature 441, 1094:1096, 2006). However, neuronal integration, differentiation and survival following transplantation are
still a bottle neck for such approaches. Our main goal is to establish cell culture models to investigate genetic and
pharmacological manipulations leading to increased neuronal differentiation and survival in cell-based therapies. Here, we study
the role of CREB (cAMP-response element binding) signaling.
Embryonic day 18 cortical cells from C57/Bl6 mice were digested in 0,05% Trypsin, dissociated and grown as primary
neurospheres in DMEM/F-12 supplemented with 2% B27, (1% Penicillin/Streptomycin, 4.500 mg/L glucose, 1% glutaMAX) 10
ng/mL basic fibroblast growth factor (FGF), and 10 ng/mL epidermal growth factor (EGF). After 7 days, primary neurospheres
were plated onto PDL-coated glass cover slips in DMEM F-12/(1% Penicillin/Streptomycin, 4.500 mg/L glucose, 1%
glutaMAX)/2% B27/1% fetal bovine serum for differentiation. Two hours after plating, cells were transfected with GFP, ACREB
(negative dominant of CREB) or CREB s142l (constitutively active) either by retrovirus or Ca2+-phosphate. To manipulate
signaling cascades involved in the activation of CREB-dependent-transcription, cultures were treated with H-89 (PKA-inhibitor),
KN-62 (CAMK-inhibitor), U0126 (MAP/ERK-inhibitor) or Wortmannin (PI3K- inhibitor) 24h after plating. Cultures were then
maintained under differentiation condition for 7-10 d before fixation with 4% PFA and immunocytochemistry for GFP and
neuronal markers (-III tubulin or MAP2). To analyze the morphology of the neurons, images were taken with an epifluorescence microscope and analyzed in Image J v1.44 (National Institute of Health). Total dendritic length, number of dendritic
branches and longer dendrites were measured. Our preliminary results indicate that Ca2+-phosphate methods is more efficient
than retroviruses to transfect neurons derived from cortical neurospheres. Dendritic morphology of cortical neurons is
significantly affected by treatment with H-89 (PKA-inhibitor), KN-62 (CAMK-inhibitor), U0126 (MAP/ERK-inhibitor) and
Wortmannin (PI3K- inhibitor), similar to what has been described following forced expression of CREB.dominant negatives
(Neuron 34, 999:1010, 2002).
Conclusions:
Our results suggest that CREB mediated transcription can be activated through different signaling pathways and is important for
dendritic differentiation of newly generated neurons.
Keywords: CREB, signaling cascades , Neurospheres, neuronal differentiation and survival, dendrites
Financial Support: FAPERN, CAPES and CNPq.
Resumo:12-038
INVESTIGATION OF PURINERGIC SYSTEM IN NON-DIFFERENTIATED AND DIFFERENTIATED STEM
CELLS
Iser, I. C. 1; Bracco, P. A. 1; Zaninn, R. F. 5; Lenz, G. 4; Nardi, N. B. 2; Battastini, A. M. O. 3; Wink, M. R. 1

1
Departamento de Cincias Bsicas da Sade, UFCSPA
2
Departamento de Gentica, UFRGS
3
Departamento de Bioqumica, UFRGS
4
Departamento de Biofisica, UFRGS
5
Faculdade de Farmcia, PUCRS
Objectives:
Extracellular nucleotides are signaling molecules, involved in important processes in the body. Recently, it has been suggested
that the purinergic system may be involved in the biology of mesenchymal stem cells (MSCs). A particular interest has been
focused on the role of this system in the proliferation and differentiation of this cells. In view of this, our goal was to analyze the
degradation rate of adenine nucleotides as well as the expression of NTPDases and ecto-5'-nucleotidase (CD73) in nondifferentiated and differentiated MSCs.
The cultures of MSCs from BALB/c mice non-differentiated and differentiated into adipocytes and osteoblasts, were submitted to
the determination of enzymatic activity for hydrolysis of ATP, ADP and AMP, by measuring the release of inorganic phosphate
by the Chan method. Protein content was performed by the Comassie blue method. The real-time PCR technique was performed
to verify the expression of NTPDases, using primers for the six types of NTPDases and CD73. Our data suggest that ATP, ADP
and AMP hydrolysis increase when the cells are differentiated in osteoblasts, however, in the adipogenic differentiation, there is
increase only in the AMP hydrolysis. Moreover, non-differentiated and differentiated MSCs express NTPDase 1,2, 3, 5 and 6 and
CD73 in similar levels.
Conclusions:
ATP may be required during the process of differentiation in osteoblasts but not necessary when differentiation is complete,
explaining its high hydrolysis after differentiation. Moreover, we believe that adenosine generated might be involved in the
processes of MSCs differentiation, since the presence of receptors for this molecule has been reported in the process of
differentiation of hematopoietic precursor cells. Finally, the presence of these ectoenzymes and the differential ectoncleotidases
activities in undifferentiated and differentiated cells suggest that these enzymes can modulate the concentration of extracellular
nucleotides and nucleosides, generating a signal that can be important in physiological regulation and maintenance of
undifferentiated and differentiated MSCs.
Keywords: stem cells, purinergic, differentiation
Financial Support: CAPES and CNPq.
Resumo:13-068
-MSH EFFECTS ON RHODOPSIN, TYROSINASE AND MC1R GENES IN B16 MUS MUSCULUS
MELANOCYTES.
Glria, T. H. R. ; Castrucci, A. M. D. L.
Departamento de Fisiologia, Instituto Biocincias, USP, IB
Objectives:
In vertebrates, skin color is given by pigments, synthesized and/or stored in cutaneous pigment cells. The vertebrate color change
is mainly regulated by -MSH and a family of melanosome enzymes, which includes tyrosinase and tyrosinase-related proteins 1
and 2 (TRP-1 and TRP-2, respectively). -MSH action is associated with melanosome dispersion and/or melanin synthesis,
processes which lead to body darkening, whereas melanin aggregation or synthesis inhibition results in skin lightening. Opsins,
such as melanopsin and rhodopsin, may be expressed in skin pigment cells, besides being present in the retina, and may mediate
non-visual photo-responses such as cell proliferation and melanosome dispersion. The aim of this study was to investigate the
temporal expression of rhodopsin, tyrosinase and of -MSH receptor MC1R, as well as the effects of 10-7 M and 10-8 M -MSH
for 24 hours in Mus musculus B16 melanocytes, kept in constant darkness.
In this work we used quantitative PCR to investigate the expression of rhodopsin, tyrosinase and the MC1R receptor genes in
Mus musculus melanocytes submitted to constant darkness regimen, in the presence or absence of -MSH. Cultured melanocytes
were seeded (1x106 cells) in 25 cm2 culture flasks, and kept at 37C in constant darkness for 2 days. Total RNA extraction was
performed every four hours along 20 hours starting at 0 hour of the third day. The samples were submitted to RT-PCR followed
by quantitative PCR for mRNA quantification of MC1R receptor, tyrosinase and rhodopsin. Using real time PCR (quantitative)
we demonstrated that 10-7 M -MSH does not modulate MC1R mRNA levels, as compared to the control group, although a
tendency to reduction was evident. On the other hand, at the concentration of 10-8 M, we observed a statistically significant
increase of the transcript level at hour 20, as compared to the control group. For rhodopsin, we demonstrated that 10-7 M -MSH
modulates mRNA levels, as compared to the control group, demonstrating a statistically significant decrease at hour 12. At 10-8
M, -MSH significantly increased the transcript levels at hour 4 and 20, as compared to the control group. The same pattern was
observed for tyrosinase, demonstrating a statistically significant decrease in the concentration of 10-7 M at hour 0 and a
significant increase in the concentration of 10-8 M at the hour 8.
Conclusions:
Using real time PCR (quantitative) we demonstrated that -MSH promotes a dose-dependent response of MC1R receptor,
tyrosinase and rhodopsin gene expression, but was not able to synchronize the expression of these genes.
Keywords: MC1R, Melanin, -MSH, Rhodopsin, Tyrosinase
Financial Support: FAPESP and CNPq grants. THRG is a fellow of FAPESP.
Resumo:13-069
AN ASSESSMENT OF THE DIABETES KNOWLEDGE AMONG HIGH-SCHOOL STUDENTS IN THE CITY OF
ITABAIANA, SERGIPE, BRAZIL
Lima, D. B. 1; Santos, D. M. 1; Camporez, J. P. G. 2; Aires, M. B. 1; Rodrigues, T. M. A. 1; Arago, J. A. 1;

Carvalho, C. R. O. 2; Maral, A. C. 1
1
Departamento de Morfologia, Universidade Federal de Sergipe, UFS
2
Dept de Fisiologia e Biofsica, Universidade de So Paulo, USP
Objectives:
Evaluate the knowledge of high school students in the city of Itabaiana-SE about diabetes, and also estimate the number of
students with the disease.
This was an exploratory descriptive study involving 712 high school students (14 to 44 years old) of the urban area in Itabaiana
City. It was applied an structured questionnaire with subjective questions covering the general knowledge about diabetes mellitus
(DM), the etiologic classification and symptom identification related to Diabetes type 1 and 2.The body mass index was
calculated based on weight and height self-reported. The results were expressed as average, confidence intervals (CI) or
percentages (p
Conclusions:
The diabetic knowledge results showed a lack of information about the disease symptoms and an ineffective comprehension of
the diabetes types and the specific symptoms.
Keywords: Diabetes, Knowledge, Students
Resumo:13-070
USE OF MARKERS OF OXIDATIVE STRESS AS INDICATORS OF INJURY OF THE LOWER EXTREMITIES IN
PATIENTS WITH TYPE 2 DIABETES MELLITUS.
Teixeira, C. J. 1; Oliveira, A. C. P. 1; Stefanello, T. F. 1; Carrara, M. A. 5; S-nakanishi, A. B. D. 2; Comar,

J. F. 2; Santos, C. A. D. 3; Mareze-costa, C. 4; Bazotte, R. B. 5; Batista, M. R. 1
1
Clinical Analysis Depart. / State University of Maring, UEM
2
Biochemistry Departament / State University of Maring, UEM
3
Statistical Departament / State University of Maring, UEM
4
Morphological Sciences Depart. / State University of Maring, UEM
5
Pharmacology Departament / State University of Maring, UEM
Objectives:
Oxidative stress, caused by an imbalance between production of reactive oxygen species and endogenous antioxidant capacity is
a major determinant of diabetic complications. We investigated the role of oxidative stress markers as indicators of risk of injury
of the lower extremities in patients with type 2 diabetes mellitus (PPDMT2).
In a group of 29 patients (the Ethics Committee of the State University of Maring - Protocol no. 381/2010) were assessed the
following parameters: 1. Anthropometric measures (weight and height) for calculating the Body Mass Index (BMI); 2. Glycated
hemoglobin (HbA1c); 3. Three blood markers of oxidative status (total antioxidant capacity - TAC, reduced proteins thiol groups
and reduced levels of protein carbonyls), and 4. Test of sensation in the feet, as determined by to the National Hansen's Disease
Program (HRSA U.S.). Patients were divided into two groups according to the index of sensation in the feet: group 1 (low risk
of injury 18 PPDMT2) and group 2 (high risk of injury 11 PPDMT2). It is noteworthy that the metabolic evaluations were
performed at two moments in each group, baseline (time 0) and later (110 days). The results were: 1. The group 1 had a mean age
56.98.7 years and the median duration of disease of 120 [0-240] months, while group 2 had a mean age of 63.47.6 years and
the median of disease duration of 180 [60-480] months (P
Conclusions:
The results showed an increase in HbA1c levels reflected in the increase of the marker pro-oxidant and reduction in markers of
antioxidant. Thus, we conclude that these markers of oxidative stress may be a good parameter to assess the degree of impairment
or worsening of vascular lesions in the lower extremities in type 2 diabetes.
Keywords: type 2 diabetes mellitus, diabetic foot, oxidative stress
Financial Support: Fundao Araucria.
Resumo:13-071
MATERNAL NICOTINE EXPOSURE DURING LACTATION ALTERS HYPOTHALAMUS- PITUITARYADRENAL FUNCTION IN MALE ADULT OFFSPRING.
Younes-rapozo, V. ; Bernardino, D. N. ; Peixoto-silva, N. ; Rodrigues, C. P. ; Santos-silva, A. P. ; de

Moura, E. G. ; Manhes, A. C. ; de Oliveira, E. ; Lisboa, P. C.
Departamento de Cincias Fisiolgicas/IBRAG, UERJ
Objectives:
We have previously showed that maternal exposure to nicotine during lactation - from postnatal day 2 (P2) to P15, causes some
metabolic disturbance in pups, such as higher serum leptin, higher catecholamine content, lower tyrosine hydroxilase expression
and higher serum corticosterone at the end of treatment (J Endocrinol 205 159-170, 2010), evidencing an acute effect of nicotine.
Our objective here was to evaluate the long-term effects of maternal nicotine exposure exclusively during lactation, specifically
in the hypothalamic-pituitary-adrenal axis, by the immunostaing of corticotropin-releasing factor (CRF) and adrenocorticotropic
hormone (ACTH) and the analyses of serum corticosterone.
At postnatal P2, dams were subcutaneously implanted with osmotic minipumps releasing nicotine (NIC - 6mg/Kg/day) or saline
for 14 days (control group). P180 male offspring (n=4/group, one animal from each litter) were intracardially perfused with 0.9%
saline solution followed by 4% paraformaldehyde and 4% paraformaldehyde plus 10% sucrose for cryoprotection. Brains and
pituitaries were sectioned at 20micrometers on a cryotome at - 20C. Hypothalamus sections were incubated with anti-CRF
(1:100 from Abcam) and pituitaries with anti-ACTH (1:100 from Santa Cruz). Immunoreaction was visualized by incubation
with secondary antibodies conjugated with Alexa Fluor 488 or Alexa Fluor 555 (1:400 from Invitrogen, Molecular Probes) and
sections were also counterstained with DAPI (1:5000 from Sigma). Serum corticosterone was measured using a specific
commercial RIA kit (ICN Biomedicals Inc) (n=6/group, one animal from each litter). NIC animals showed a higher CRF
immunostaining, and CRF-positive fibers were apparently thicker but smaller, in comparison to control group. In pituitaries, NIC
male offspring showed ACTH-positive cells with increased immunoreactivity in comparison to control animals. Concerning
serum corticosterone, NIC group presented higher levels (+77%, p
Conclusions:
Our results suggest that the adrenal dysfunction of male offspring in early life by maternal nicotine exposure can alter
permanently the hypothalamic-pituitary-adrenal axis at adulthood, stimulating all the axis, reinforcing the idea that maternal
smoking during lactation can be a risk factor for development of later hormonal disorders.
Keywords: nicotine exposure, lactation, CRH, ACTH, corticosterone
Financial Support: CAPES, FAPERJ, CNPq.
Resumo:13-072
GHRELIN SIGNALING IN ADIPOSE TISSUE OF OBESE MICE IS ALTERED DURING THE DEVELOPMENT.
Soares, V. . M. ; Garcia-souza, E. P. ; Miranda, G. L. ; Neves, F. A. ; Bernado, A. F. ; Gomes, I. A. ; Lessa,

J. G. ; Moura, A. S.
Instituto de Biologia Roberto Alcantra Gomes - UERJ, IBRAG
Objectives:
Human overnutrition has caused a rise in the prevalence of obesity in recent years. In addition to the deleterious effects of obesity
during childhood, the long-term effects in adulthood have also been described. Intensive efforts are underway to clarify nutrienthormone interactions contributing to weight gain. The gastric peptide hormone ghrelin, an endogenous ligand for GHS-R, has
generated considerable interest as an important stimulus for weight gain and control body fat mass by modulating the storage and
oxidation of fatty acids. This study aims to evaluate the association of ghrelin signaling in adipose tissue in mice overfed during
lactation after weaning and during adulthood.
To induce early postnatal overnutrition, Swiss mice were overfed through litter size reduction. In this model, the litter was
adjusted to only three male pups per litter three days after birth (overfed group; OG). For the control group, the litter size was
adjusted to nine newborns (CG).The CG and OG were studied at 21 and 180 days. We evaluated anthropometric parameters as
body mass and visceral adipose tissue. Fasting glucose was measured using glucometer and test strips. Histological sections of
adipose tissue were paraffin embedded and stained with hematoxylin and eosin to determine the area and diameter of adipocytes,
using Olympus BX40 microscope and analyzed with Image-Pro Plus software version 4.5.1. Analysis of the protein content of
GHS-R 1a, CPT1, PPAR, UCP2, AKT, pAKT, PI3K and actin were detected by Western blotting method. Statistical analysis was
performed using Student's t-test and values expressed as mean and standard error. Results: In these animals, overfeeding during
lactation was able to induce a significant increase in body weight starting at the 21th day of life, and this increased weight
persisted until 180 days of age (21 days- CG: 13.43 0.76, OG: 19.57 0.29 g, N=8, 180 days - CG: 48.15 0.85, OG: 67.8
1.9 g, n = 17). We observed a significant increase in fasting glucose between groups at 180 days (C: 114.0 9205, O: 160.2 5.9
mg / dl), and of visceral adipose tissue weight at 21 days (CG: 0.03 0.005 g, N=6, OG: 0.25 0.05g N=6) and 180 days(CG:
2.3 0.67 , OG: 5.31 0.37 g, n = 8) compared to CG animals. OG Adipocytes showed a larger area (CG: 3095.7 484, OG:
6292.9 902 m2, n = 5) and diameter (CG: 75.7 1.85, OG: 104.7 4 , 0 mM, n = 5) compared with CG. At 21 days of age OG
presented a expressive increase in the GHSR1a (CG: 18,330 4433, OG: 44,460 1778 AU, n = 5) and PI3K content (CG:
14,560 1355, OG: 39,620 5920 AU n = 6) in adipose tissue. However, this profile has changed in adulthood, since GHSR1a,
PI3K, AKT, pAKT and PPAR gamma content were reduced in adipose tissue of OG compared to CG at 180 days old.
Conclusions:
Overfeeding during lactation induces obesity, increased visceral fat mass, adipocyte hypertrophy. Ghrelin receptor expression and
key signaling proteins of the hormone action were found altered. The data suggested an association of the hormone with storage
and oxidation of fatty acids during mice developme
Keywords: Ghrelin, overnutrition, adipose tissue
Financial Support: CNPq, Capes, FAPERJ, Vital Brasil
Resumo:13-073
LONG-TERM EFFECTS OF NEONATAL HORMONAL MANIPULATION ON BONE MINERAL AND
BIOMECHANICAL PROPERTIES OF FEMUR RATS
Mello, W. G. D. 1,4; Morais, S. R. L. D. 1,4; Biffe, B. G. 1,4; Louzada, M. J. Q. 2; Dornelles, R. C. M. 4;

Nakamune, A. C. D. M. S. 4; Antunes-rodrigues, J. 3; Bedran de Castro, J. C. 4
1
Multicentric Graduate Studies Prog. in Physiological Science, SBFis
2
Department of Support to Production and Health Animal , FMVA-UNESP
3
Department of Physiology, FMRP-USP
4
Department of Basic Sciences , UNESP - Araatuba-SP
Objectives:
Several functions wich are controlled by the brain are expressed differently in male and female mammals, including the central
regulation of bone mass. Therefore, the aim this study was analyze the long-term changes in bone mineral and biomechanical
properties induced by sexual dimorphism in the hypothalamic control of bone mass
Wistar rats, newborns, were divided into four groups (n = 08 per group). Male pups were cryoanesthetized and castrated or
sham/castrated by 24 hours after birth; female pups from separate litters were injected SC with testosterone propionate 100 g in
50L corn oil (androgenized female) or oil vehicle 50L (control female) and were observed for 20, 40 and 120 postnatal day,
both femurs were collected and preserved in physiological saline at -20 C by the day of experiments. The length and thickness of
the left femurs were performed with aid of a caliper; after measuring, they were dried overnight at 90 C and ashed at 900 C for
24 h for determined the mineral content. The bone mineral density (BMD) was determinate in the right femurs by dual-energy Xray absorptiometry (DXA) and, the mechanical properties were assessed by Three-Point Bending Testing; these data were used
for the acquisition and calculation of the structural properties: ultimate strength (N), toughness (N.mm), and stiffness (N/mm).
Data were analyzed using ANOVA and Tukey and expressed as the meansSE. The results showed that bone mineral content and
biomechanical properties in all groups increased rapidly with aging. However, the neonatal exposed to androgen induced changes
in growth, BMD and bone mass quality in androgenized females, leading to a masculinization pattern in development. On the
other hand, neonatally castrated males had the bone development and quality of the mechanical properties similar to those control
females.
Conclusions:
In conclusion, our work indicated that the long-term organizational influence of the neonatal castration and androgenization,
which is known inducing irreversible effects on CNS and hypothalamus, also induced a long-lasting modification in bone
structure and strength. These results provide new insight into the dynamic complexity of bone mass homeostasis.
Keywords: ANDROGEN, BONE PROPERTIES, SEXUAL DIMORPHISM
Resumo:13-074
DPP-IV ACTIVITY AND INFLAMMATORY BIOMARKERS IN TYPE 2 DIABETES MELLITUS
Bell, L. P. 1; Bitencourt, P. E. R. 2; Bona, K. S. 2; Moretto, M. B. 2

1
Departamento de Anlises Clnicas e Toxicolgicas , USP
2
Departamento de Anlises Clnicas e Toxicolgicas, UFSM
Objectives:
Type 2 diabetes mellitus (type 2 DM) is one of the worlds most important diseases and is associated with a general activation of
the innate immune system, in which there is a chronic state of low-grade inflammation (Trends Endocrinol. Metab. 19; 10, 2008).
Dipeptidyl peptidase IV (DPP-IV, CD26) is a glycoprotein widely expressed in lymphocytes surface, where it is binding with
adenosine deaminase (ADA) (J. Immunol, 156; 1349, 1996). Moreover, DPP-IV has been proposed as a target for
pharmacological intervention in patients with type 2 DM (Curr. Diabetes Rev. 5; 92, 2009). Thus, the aim of this study was to
investigate CD26 expression and activity as well as ADA and n-acetyl--d-glucosaminidase (NAG) activities in lymphocytes
from type 2 DM and control subjects.
We obtained lymphocytes from 32 fasting type 2 DM patients (HbA1c= 7.36 0.22%) and 23 fasting control subjects (HbA1c=
5.00 0.17%). In lymphocytes we determinated DPP-IV, ADA and NAG activity and CD26 expression. The results were
described in mean S.E. We observed an increase in ADA (C=7.750.49; type 2 DM=9.370.51U/L) and NAG (C=23.632.5;
type 2 DM=29.861.4mmol/L) activities in lymphocytes from type 2 DM when compared with control subjects. DPP-IV activity
(C=83.396.5; type 2 DM=89.124.5U/L) did not alter between the groups and the CD26 expression decrease in type 2 DM
patients (C=14.681.7, type 2 DM=8.961.2%). Moreover, we observed a positive correlation between ADA and NAG activity
(R= 0.4272; P
Conclusions:
We conclude that in type 2 DM there is no coestimulatory activity (low CD26 expression), possible because the no alteration in
DPP-IV activity. The increase in ADA and NAG activity indicate an inflammatory state in these patients. This result is reinforced
by the positive correlation between NAG and ADA activity and the negative correlation between NAG and CD26 once it is
known that there is an inverse relationship between CD26 and inflammation.
Keywords: type 2 diabetes mellitus, lymphocytes, dipeptidyl peptidase IV, adenosine deaminase, n-acetyl-b-d-glucosaminidase
Financial Support: Coordenao de Aperfeioamento de Pessoal de Nvel Superior (CAPES)
Resumo:13-075
GENOTYPIC DISTRIBUTION OF THE FAT MASSAND OBESITY-ASSOCIATED (FTO) GENE
POLYMORPHISMS AND ASSOCIATION WITH C-REACTIVE PROTEIN AND METABOLIC VARIABLES IN
RECENT POSTMENOPAUSAL WOMEN RECEIVING HORMONE THERAPY
Ramos, R. B. ; Casanova, G. K. ; Spritzer, P. M.

Gynecological Endocrinology Unit, UFRGS/HCPA, GEU, UFRGS/HCPA
Objectives:
Postmenopausal women are at a higher risk of hypertension, unfavorable changes in the lipoprotein profile, diabetes, and
cardiovascular disease as compared with their premenopausal counterparts. Hormone therapy (HT) seems to have different
impact on cardiovascular (CV) risk, according to the age and time after menopause and ultra sensitive C reactive protein (usCRP)
has been regarded as a good marker in predicting this CV risk. The fat massand obesity-associated gene (FTO), located in the
chromosome 16, is associated with fat mass and body mass index (BMI), risk of obesity, metabolic syndrome and high CV risk.
The present study aimed to assess the genotypic distribution of the single-nucleotide polymorphisms (SNPs) rs9939609 T>A and
rs8050136 A>C located in the intron 1 of the FTO gene and verify whether these gene variants are associated with usCRP and
metabolic variables in recent postmenopausal women receiving HT.
Sixty-six participants (51 3 years, 22 10.2 months since menopause) consulting for symptoms of estrogen deficiency were
clinically evaluated before and after 6 months of HT. DNA was extracted from peripheral blood by a standard salting out
procedure and genotyped by allelic discrimination assay with Real Time PCR. The genotypic distribution of the SNPs rs9939609
(TT: 47%, TA: 42.4%, AA:10.6%) and rs8050136 (AA: 13.6%, AC:34.9%, CC: 51.5%) were similar to previous studies. During
HT, waist circumference and systolic blood pressure were reduced. Women with the homozygous polymorphic AA genotype of
the SNP s9939609 showed a decrease in usCRP [2.1 (0.9 -4.8) to 1.2 (0.5 -1.9) mg/L, p= 0.02] and triglycerides [158.716.9 to
129.122.3 mg/dL, p=0.02] after HT, while no changes were observed in women with the TT+TA genotypes. In contrast,
considering the SNP rs8050136, only women with the wild AA genotype presented a decrease in the usCRP [1.84 (0.77 -4.49) to
1.23 (0.61-2.23) mg/L, p0.008] after HT, but not those with the polymorphic AC+CC genotypes. Anthropometric and other
metabolic variables remained unchanged before and after HT in both polymorphisms.
Conclusions:
Our results indicate that these FTO gene variants may exert dissimilar influence on the effects of HT: the wild genotype of the
SNP rs8050136 and the homozygous polymorphic genotype of SNP rs9939609 presenting a beneficial impact on usCRP. These
data suggest a possible linkage desequilibrium in the 1 and 2 intronic regions of the FTO gene.
Keywords: FTO gene, Hormone Therapy, Menopausal Women, Polymorphisms
Financial Support: CAPES, INCT Hormones and Womens Health- CNPq
Resumo:13-076
ENDOCRINE AND METABOLIC FUNCTIONS ARE ALTEREDED IN A MODEL OF GENERALIZED ANXIETY
DISORDER
Mousovich-neto, F. 1; Lonreno, A. L. 1; Landeira-fernandez, J. 2; Corra-da-costa, V. M. 1

1
Instituto de Biofsica Carlos Chagas Filho/UFRJ, IBCCF/UFRJ
2
Departamento de Psicologia/Pontifcia Universidade Catlica, PUC
Objectives:
Freezing in response to threat is used by many species as an adaptative-defensive strategy. Carioca High-Freezing rats (CHF) are
animals selected by their high freezing response in contextual fear conditioning and represent a great model of generalized
anxiety disorder. Here we evaluated some aspects of endocrine and metabolic functions in those animals, by evaluating
corticosterone and testosterone serum levels, thyroidal function, serum colesterol and triglycerides, fasted glucose serum levels,
glucose tolerance, oxigen consumption and epididymal and retroperitoneal fat depots weight.
Control male Wistar rats (C) and CHF animals were sacrificed by decaptation and blood was collected and centrifuged at 1200x
g, to obtain serum. Corticosterone, testosterone and thyroid hormones were measured by specific radioimunoassays, and
colesterol and triglycerides through an enzymatic-colorimetric assay. Oxygen consumption was measured using an air
composition analyzer. Epididymal and retroperitoneal fat depots were removed and weighted. Glucose tolerance test were
performed in fasted animals by administration of 1.83 x 10-3 mol/glucose/100g of body weight and glycemia measurements were
done 30, 60, 120 and 180 minutes after the glicemic orogastric infusion. Fast glycemia was considered the first measure of the
glucose tolerance test. Serum corticoesterone was higher in CHF animals (C: 118.927.97 vs CHF: 339.049.38 ng/mL), while
serum testosterone was decreased (C: 3.330.294 vs CHF: 2.030.29 ng/mL). Serum T3 is decreased (C: 58.402.401 vs CHF:
46.952.846 ng/dL) while T4 and thyrotropin serum levels were unaltered in CHF animals. Serum colesterol was higher if
compared with control group (C: 181.65.61 vs CHF: 226.413.04 mg/dL) as well as serum triglycerides (C: 41.46.03 vs CHF:
82.217.4 mg/dL). Fat depots were significantly higher in CHF animals; epididymal (C: 2.40.26 vs CHF: 4.30.38 g) and
retroperitoneal (C: 1.80.212 vs CHF: 3.80.58 g). Oxygen consumption was lower in CHF animals when compared to control
animals (C: 10.55 vs CHF: 7.95 VO2 ml/min/kg0.75). Fast glucose was higher in CHF animals (C: 68.73.04 vs 82.32.9
mg/dL) however there was no difference between groups in glucose tolerance test.
Conclusions:
In CHF animals, anxiety disorder induces important endocrine and metabolic disfunctios
Keywords: anxiety, endocrine disorders, metabolic disorders
Financial Support: CNPq, FAPERJ, CAPES and SBEM Thyroid Department.
Resumo:13-077
SODIUM APPETITE AFTER SODIUM RESTRICTION: AN INTEGRATIVE VIEWPOINT
Mecawi, A. S. 1,2; Fonseca, F. V. 2; Vilhena-franco, T. 1; Reis, L. C. 2; Elias, L. L. K. 1; Antunes-rodrigues,

J. 1
1
Department of Physiology, FMRP-USP
2
Department of Physiological Sciences , UFRRJ
Objectives:
Salt intake is a behavior implicated in the integrative regulation of extracellular volume, plasma tonicity and blood pressure and is
directly influenced by changes in sodium homeostasis. Sodium dietary restriction is an interesting noninvasive model to study the
sodium appetite. Our aim was to investigate the integrative control of salt and water intake after dietary sodium restriction.
To this purpose, Wistar male rats received normal- (1% NaCl) or low- (< 0.01) associated with a decrease in water intake (9.6
0.8 mL vs. 13.1 0.5 mL, p < 0.05) and a decrease in the extracellular volume as demonstrated by increase in hematocrit (42.8
0.7 % vs. 38.8 0.6 %, p < 0.01) and plasma protein (7.0 0.1 mg/dL vs. 5.7 0.1 mg/dL, p < 0.001). Additionally the low-Na+
diet decreased urinary volume (5.4 0.6 mL vs. 7.6 0.5 mL, p < 0.05) and sodium excretion (0.26 0.01 mE/100g/day vs. 1.66
0.07 mE/100g/day, p < 0.01). We observed a discrete decreased arterial pressure (101 1.5 mmHg vs. 105 1.1 mmHg, p <
0.05) without effect in heart rate. There was no effect of low-Na+ diet on plasma atrial natriuretic peptide and vasopressin
concentrations. An increase in plasma angiotensin II (119 12 pg/mL vs. 39 11 pg/mL, p < 0.001) and decrease in plasma
oxytocin (0.60 0.04 pg/mL vs. 0.99 0.12 pg/mL, p < 0.05) concentrations after low-Na+ diet was observed. These animals
also presented an increase in 1.8% NaCl intake at 30 min (0.44 0.05 mL vs. 0.06 0.02 mL, p < 0.05) and 5 hours (1.65 0.13
mL vs. 0.57 0.05 mL, p < 0.001) after four days of low-Na+ diet. Associated to this, low-Na+ diet group also presented sodium
intake-dependent increase in water intake at 1 hour (0.99 0.09 mL vs. 0.49 0.12 mL,p < 0.05) and 5 hours (2.86 0.13 mL vs.
1.64 0.21 mL, p < 0.001). Regarding the sodium appetite, the low-Na+ diet induced an important increase in sodium preference
at 30 min (31.4 3.5 % vs. 12.9 4.8 %, p < 0.001) and 5 hours (34.1 1.9 % vs. 22.1 1.9 %, p < 0.01).
Conclusions:
Our results indicate that sodium appetite control and the strong increase in sodium preference after sodium restriction is
dependent on a variety of factors, that could activate sodium appetite signals (volume/ baroreceptors and peripheral Ang II) and
inhibition of sodium satiety signals, as oxytocin.
Keywords: renin-angiotensin system, oxytocin, low sodium-diet, salt intake
Financial Support: CNPq/FAPESP
Resumo:13-078
ENDOCANNABINOID SYSTEM AND INFLAMMATORY RESPONSE IN ADIPOSE TISSUE DURING OBESITY.
Gotardo, . M. F. ; Santos, A. N. ; Ribeiro, M. L. ; Gambero, A.

Clinical Pharmacology and Gastroenterology Unit, USF
Objectives:
Obese animals and humans over activated the endocannabinoid system (CB1 and CB2 receptors, endogenous ligands and
enzymes for synthesis or degradation of ligands) in several tissues involved in metabolic process, including adipose tissue (AT).
The endocannabinoid system also modulates inflammatory response, mainly through CB2 activation in inflammatory cells. Thus,
we investigated if endocannabinoid system expression is altered in AT and in macrophages infiltrated in AT from high-fat diet
mice. We also analyzed the CB2 cannabinoid agonist action in macrophage/adipocyte co-culture system, an in vitro model of
inflammation in adipose tissue.
Swiss mice were feed with high-fat diet (HFD) or standard diet (N) during 12 or 24 weeks (n=12/group). Body weight and
glucose homeostasis (glucose basal and Insulin test tolerance) were evaluated. Epididymal adipose tissue was collected for gene
expression analysis by qRT-PCR (CB1, CB2, N-acyl-phosphatidylethanolamine (NAPE), fatty acid amide hydrolase (FAAH),
diacylglycerol lipase (DAGL)). 3T3-L1 adipocytes and RAW 264.7 macrophages were co-cultured in presence or absence of
CB2 agonist JWH-015 (1, 3 e 10 M). TNF-&alpha was measured by ELISA in co-cultures stimulated or not by
lipopolysaccharide (LPS, 1 ng/ml). HFD mice presents high body weight (41.31.1 and 56.81.8 g for C and HFD 12 weeks;
45.51.2 and 622.2 g for C and HFD 24 weeks) and insulin resistance (3.380.75 and 1.170.09 for C and HDF 12 weeks;
1.790.6 and 0.5020.09 for C and HFD 24 weeks), confirming the obesity status. CB2 receptor gene expression in AT was
higher in obese mice after 12 or 24 weeks (14.03.3 and 3.30.4 arbitrary unit (AU) for HFD and C 12 weeks group,
respectively; 19.82.1 and 11.32.0 AU for HFD and C 24 weeks group, respectively). NAPE expression was reduced only after
24 weeks in AT (15.54.2 and 38.85.8 AU for HFD and C 24 weeks, respectively). When isolated macrophages were analyzed
no differences were observed in CB2 or NAPE expression. Basal or LPS-stimulated TNF-&alpha release in the co-culture system
were reduced by 10M JWH 015 (1243,260,4 e 982,0141,9 pg/ml in the absence and presence of JWH 015 after LPS
stimulus).
Conclusions:
Increased CB2 expression observed in AT from obese mice could be due an increased macrophage infiltration, because in
isolated macrophage these alterations were not detected. Anyway, the stimulation of CB2 receptor reduces TNF-&alpha
production by adipocytes/macrophages suggesting an anti-inflammatory action that could be very important in the control of AT
inflammation during obesity.
Keywords: inflammation, macrophages, adipocytes, CB2 receptors, adipokines
Resumo:13-079
MATERNAL HIGH-FAT DIET DURING PREGNANCY AND LACTATION PROGRAMS THE OFFSPRING FOR
OBESITY AND LEPTIN RESISTANCE IN MICE.
Volpato, A. M. ; Magalhes-da-costa, E. ; Monterlei, R. K. S. ; Aguila, M. B. ; Mandarim-de-lacerda, C. A.

; Correia, M. L.
Departamento de Anatomia , UERJ
Objectives:
Fatty diet during pregnancy in rat dams programs for metabolic syndrome (MS) in the offspring. We tested the hypothesis that
the offspring of dams fed high fat diet during pregnancy and lactation develops MS and leptin resistance.
Pregnant C57BL6 mice (n=20) were fed either standard chow (SC; 19% fat) or high fat diet (HF; 49% fat). After weaning, male
offspring was divided in four groups according to diet of dams and offspring: SC(dams)/SC(offspring), SC/HF, HF/SC and
HF/HF (n=12/gp). MS was characterized by weight gain curve measured weekly (g); tail-cuff systolic pressure (mmHg), areas
under the curve after oral glucose tolerance test (OGTT) and fat mass depots performed at 12 weeks of age (g). To analyse leptin
sensitivity, each group was divided into two groups (vehicle or leptin-5g) to verify the feeding response (Kcal) after acute
intracerebroventricular (ICV) treatment (n=6/gp). Offspring born from HF dams does not present higher body mass at birth and at
weaning (data not shown). However, at 6 weeks of age, HF/HF animals were heavier than SC/SC (+10%, p
Conclusions:
In conclusion, HF diet during pregnancy and lactation programs the offspring to the development of obesity. Patterns of blood
pressure and glucose metabolism can be affected only if HF diet persists after weaning. Importantly, HF diet during gestation and
lactation; only after weaning or during whole life alter the feeding response to leptin, which could mechanistically contribute to
the development of obesity.
Keywords: obesity, programming, leptin resistance
Financial Support: Cnpq, FAPERJ
Resumo:13-080
TYPE 2 IODOTHYRONINE DEIODINASE ACTIVITY IS INHIBITED BY THIMEROSAL
Pantaleo, T. U. ; Padrn, A. S. ; Ferreira, A. C. F. ; Carvalho, D. P. D. ; Rosenthal, D. ; Guimaraes, J. R. D.

; Costa, V. M. C. D.
INSTITUTO DE BIOFSICA CARLOS CHAGAS FILHO/UFRJ, IBCCF/UFRJ
Objectives:
Sodium ethyl mercury thiosalicylate (thimerosal) is an ethylmercury (Et-Hg)-containing preservative that has been used for over
80 years as an antimicrobial agent in vaccines to prevent contamination. Until the removal of thimerosal from most pediatric
vaccines in 2001, the largest human exposure in the US was in children under 18 months of age. Prior to 2001, a child may have
received a cumulative dose of over 200 mg/kg in the first 18 months of life. In the body, ethylmercury can be converted to
inorganic mercury, which then accumulates in the kidney and brain. Thyroid hormones, triiodothyronine (T3) and thyroxine (T4),
play an essential role in the central nervous system development. In the brain, the T3, the metabolic active hormone is locally
produced by T4 deiodination mediated by type 2 iodothyronine deiodinase (D2). So, D2 plays an important role in adequately
maintaining local T3 levels in the brain and consequently normal brain development. We aimed to examine the effect of
thimerosal on D2 activity.
We obtained brown adipose tissue, hypothalamus and hippocampus of adult female Wistar rats. D2 activities was determined by
quantification of the radioiodine released by 125I-T4 under standardized conditions, and expressed as fmol T4 min-1.mg.ptn-1.
D2 inhibition was determined, in the presence of different concentrations of thimerosal (6mM; 1,2mM; 0,6mM; 0,2mM; 100M).
In vivo effect of thimerosal was also evaluated in female rats treated with two different doses of thimerosal (0,25 g/100g BW or
250g/100g BW, i.m.), administered twice a week for one month. Animals were sacrificed and hypothalamus, pituitary,
cerebellum and hyppocampus were obtained and processed for determination of D2 activity. Thimerosal was able to completely
inhibit the enzyme in brown adipose tissue, hypothalamus and hippocampus. The activity was reduced by 50% (IC50) in the
thimerosal concentration of 0.47 mM in BAT, 0,19mM in hypothalamus and 0,25mM in hippocampus. Despite of the clear
inhibition shown in vitro, in vivo treatment inhibited D2 activity in pituitary (C: 3.060.178 vs 2.620.140 or 2.3580.082 fmol
T4 min-1.mg.ptn-1 in a dose of 25 g/100g BW and in a dose of 250g/100g BW, respectively), but no statisticaly differeces
were detected in hippocampus, cerebellum, or hypothalamus tissues.
Conclusions:
Our data show that thimerosal is able to inhibit D2 activity both in vitro and in vivo, although the inhibition was not observed in
all tissues studied. Since this enzyme is important for local T3 generation, our results indicate a possible deleterious effect of
thimerosal, especially in the higher dose. On the other hand, there must be defense mechanisms against the effect of thimerosal
on D2 activity, since important targets of T3, such as cerebellum, hypothalamus and hyppocampus, showed no inhibition of D2
by this drug.
Keywords: DEIODINASE ACTIVITY, THIMEROSAL, THYROIDAL HORMONE METABOLISM
Financial Support: PRONEX/FAPERJ, FAPERJ, CNPq, MCT
Resumo:13-081
HIGH CONCENTRATIONS OF RESISTIN AND TNF- MAY RESULT IN REDUCED INSULIN SIGNAL IN RATS
CHRONICALLY TREATED WITH NAF
Silva, V. C. ; Chiba, F. Y. ; Shirakashi, D. J. ; Colombo, N. H. ; Garbin, C. A. S. ; Sumida, D. H.

Department of Basic Sciences, FOA - UNESP
Objectives:
The excessive (F) fluoride intake can lead to acute or chronic poisoning, such as dental fluorosis and disturbances in glucose
homeostasis. Further studies from our laboratory have demonstrated that chronic treatment with NaF causes insulin resistance
(IR) and decreased insulin signal (IS). In the literature there are several studies that correlate increased concentration of resistin
and TNF- with IR and decreased IS. Knowing that chronic treatment with F can interfere with IR and IS, the aim of this study
was to determine whether alterations in concentrations of resistin and TNF- could be involved in the reduction of IS in rats
treated with F. In order to do that, the experiment was carried out to evaluate: 1) the IRS-1 serine phosphorylation status in
epididymal white adipose tissue (WAT), 2) the plasma concentrations of resistin and TNF- in rats.
Four-week-old male Wistar rats (32 animals) were castrated. After 30 days this castration, the animals were divided into two
groups: 1) control group (CN); 2) fluoride group (FN), which was submmited for treatment with NaF (3.1 mg F / kg b.w.) in
drinking water for 42 days. After 6 weeks, the animals were subjected to 14-hour fast and then they were anesthetized to: 1)
quantify the IRS-1 serine phosphorylation status after insulin stimulation in WAT by the method of "western blotting", 2) collect
the blood samples for evaluation of plasma concentrations of resistin and TNF- by ELISA method. The results showed that the
FN group, compared to the CN group, showed a significant increase (p
Conclusions:
The increase in plasma concentrations of resistin and TNF- may have caused the decrease in IS, because an increase in the IRS-1
serine phosphorylation status in WAT was observed. Based on this, it is recommended to reduce the fluoride concentration (1100
ug F / g) in toothpaste used mainly by children with diabetes, as the ingestion of toothpaste containing fluoride can lead to
worsening in the health of these children. Research "in vitro" (Caries Res 41: 263, 2007) using a more acidic toothpaste (pH =
4.5) with 412 and 550 ug F / g has observed that these acidic toothpastes had the same effectiveness of neutral toothpaste (pH =
7.0) with 1100 ug F / g. These toothpastes (acidified with low fluoride concentration) would be a great option for young children
who often take in large amounts of toothpaste, with a major impact on the health of diabetic children.
Keywords: Diabetes Mellitus, Fluorine, Fluoride Poisoning , Insulin, Insulin resistance
Resumo:13-082
ANALYSIS OF THE OXYTOCIN EFFECTS ON BONE REMODELING IN SENILE RATS
Colli, V. C. 1; Spritzer, P. M. 2; Dornelles, R. C. M. 1

Multicentric Studies Graduate Prog in Physiological Sciences, UNESP
2
Department of Physiology,, UFRGS
3
Department of Basic Sciences, UNESP
Objectives:
Treatments for osteoporosis aim increase the bone density, acting in the decreased of reabsorption or increased the bone
formation. Studies have been developed to find new therapy to bone formation and oxytocin has been suggested as anabolic bone
mass regulator. The aim of this study was to verify the oxytocin action on bone metabolism in senile rats by analysis of alveolar
bone repair as well as the concentrations of markers of cellular activity.
Female Wistar rats (24-mo-old) in the diestrus cycle phase were divided into two groups (8 rat in each group). Two i.p. injections
of saline (NaCl 0.9%-control group) or Oxytocin (45 g/rat- treated group) was given 12 h apart. Seven days after, the right
maxillary incisor was extracted. At 35 days after oxytocin treatment the blood was collected and the right maxilla was removed.
The plasma concentrations of calcium, phosphorus and alkaline phosphatase were determined for spectrophotometry (Labtest
kits, So Paulo, Brazil). Serum osteocalcin (OC) and tartrate resistant acid phosphatase (TRAP) levels were determined (ELISA USCN Life Science Inc.). Tests were performed in duplicate samples. Middle thirds stained sections of the of the alveolus were
examined by light microscopy under 10X objective lenses, and images were obtained with a digital camera (JVC TK-1270 Color
Video Camera) mounted on the microscope, and analyzed with Leica Qwin Color/RGB software. Differences between groups
were tested using a Student t-test as the criteria of normal and equal variances between groups was met with a significance level
of 0.05 using GraphPad Prism 3.02 (GraphPad Software, Inc.; La Jolla, CA, USA). All data are reported as means with their
standard errors of the mean (SEM). The plasma concentration of calcium (control = 10.70 0.6580; treated = 10.41 0.6663
mg/dL) and phosphorus (control =4.880 0.3216; treated = 5.629 0.3932 mg/dL) was not significantly changed by treatment
with oxytocin, however, the ALP (control = 73.20 4.510; treated =130.8 11.37 U/L) and OC (control = 0.7280 0.02956;
treated = 1.117 0.04839 ng/mL) serum level was significantly higher in the treated group. The TRAP serum level (control =
2.404 0.3724; treated = 0.5867 0.07200 U/L) was significantly decreased after oxytocin treatment. Histomorphometric
analysis (control = 29.95 1.807; treated = 44.36 2.697 %) showed higher areas of bone formation in treated group.
Conclusions:
This results evidence the peripheral action of oxytocin in bone metabolism of the senile females and suggests involvement of this
hormone in decreased bone resorption.
Keywords: bone remodeling, oxytocin, senile rats
Resumo:13-083
ARGININE INDUCES GH GENE EXPRESSION BY ACTIVATING NOS/NO SIGNALLING IN RAT ISOLATED
HEMI-PITUITARIES
Olinto, S. C. F. 1; Gomes, M. A. 2; Barbosa, T. D. C. 3; Silva, F. G. D. 3; Nunes, M. T. 3

1
Faculdade De Cincias Integradas do Pontal, UFU
2
Depart.of Animal Morphology and Physiology , UFRPE
3
Department of Physiology and Biophysics, ICB-USP
Objectives:
Aim: The amino acid arginine (Arg) is a recognised secretagogue of growth hormone (GH), and has been shown to induce GH
gene expression. Arg is the natural precursor of nitric oxide (NO), which is known to mediate many effects of Arg, such as GH
secretion. Arg was also shown to increase calcium influx in pituitaries cells, which might contribute to its effects on GH
secretion. Although the mechanisms involved in Arg effects on GH secretion are well established, little is known about them on
the control of GH gene expression. This study aimed to investigate whether the NO pathway and/or calcium are involved with the
Arg effects on GH gene expression in rat isolated pituitaries.
Methods and Results: Pituitaries of Wistar rats weighing approximately 250 g were removed, divided in two halves, pooled and
incubated or not with Arg as well as with different pharmacological agents, to attain this purpose. It was observed that Arg 15
mg/ml; NO donor SNP - sodium nitroprusside: 10-3 and 10-4 M; and cGMP analogue 8-Br-GMPc 10-3M increased GH mRNA
expression 60 min afterwards: Control (C) 1,00 0,17 vs Arg 15 mg 1,90 0,30 , p
Conclusions:
Conclusion: The present study increases the body of evidence showing that Arg induces GH gene expression in hemi-pituitaries
and further demonstrates that NOS/ NO signalling pathway and calcium are essential for this effect.
Keywords: Arginine, Calcium , cGMP, GH mRNA , Nitric Oxide/NO sintase
Financial Support: Fundao de Amparo Pesquisa do Estado de So Paulo - FAPESP.
Resumo:13-084
VENTRICULAR T3-INDUCED HYPERTROPHY IS ACCOMPANIED BY SIRTUIN1 AND PPAR ALPHA PROTEIN
EXPRESSION DOWNREGULATION
Oliveira, L. S. ; Cordeiro, A. ; Souza, L. L. ; Pazos-moura, C. C.

INSTITUTO DE BIOFSICA CARLOS CHAGAS FILHO, UFRJ
Objectives:
Sirtuin1 (SIRT1) is a NAD+-dependent deacetylase that acts on transcription factors, modifying their activity. SIRT1 have
similar roles as the nuclear receptor peroxisome proliferator-activated receptor alpha (PPAR) on the metabolism regulation. In
the liver, increased SIRT1 and PPAR protein contents are normally observed in fasting, an important response for adaptation to
deprivation conditions. PPAR activation increases SIRT1 expression, whereas SIRT1 knockout mice have disturbed PPAR
transcriptional activity. It was described that SIRT1 and PPAR are downregulated in cardiac hypertrophy models by
isoproterenol administration or pressure overload, and these reports characterized them as part of protection response to
hypertrophy. Since thyroid hormone status variation promotes structural cardiac changes, we aimed to investigate SIRT1 and
PPAR expression in ventricle of hypo- and hyperthyroid mice, and its possible correlation with thyroid hormone-induced
hypertrophy.
Male mice (2 months of age) were rendered hypothyroid (HYPO) by addeding propyl-thyouracil 0.15% in the chow during 4
weeks and, hyperthyroidism (HYPER) was induced by T3 subcutaneous daily injections for 14 days (50 ug/100g body weight).
These mice were compared to euthyroid littermates (EU). Another experimental protocol was developed to compare hormone and
food restriction effects. A group of HYPER mice received the same food amount as EU mice consumed, as well as, a group of
EU animals were submitted to equal percentage of restriction as HYPER restricted, in mean 40% restriction during 13 days.
Ventricular SIRT1 and PPAR protein content were analyzed by Western blotting. One-way ANOVA followed by the StudentNewman-Keuls test was used for comparisons between different groups and, differences were considered significant at p
Conclusions:
We demonstrated that SIRT1 and PPAR protein levels in the cardiac ventricle are downregulated in hyperthyroidism and
upregulated in hypothyroidism. These results suggest that SIRT1 and PPAR could be involved in thyroid hormone effects on
ventricular mass.
Keywords: THYROID HORMONE, HYPERTROPHY, SIRTUIN1, PPAR ALPHA
Financial Support: CNPq, CAPES and FAPERJ
Resumo:13-085
EVALUATION OF GADD45 AND MEG3 EXPRESSION BY QUANTITATIVE REAL TIME PCR IN SPORADIC
PITUITARY ADENOMAS.
Pesce, F. G. ; Mezzomo, L. C. ; Gonzales, P. H. R. ; Kretzmann, N. A. ; Oliveira, M. C. ; Kohek, M. B. F.

Objectives:
This study evaluated MEG3 and GADD45 gene expression by qRT-PCR in sporadic functioning and clinically non-functioning
human pituitary adenomas morphologically characterized by immunoistochemical analysis.
Fourteen clinically nonfunctioning, 10 GH-secreting, 9 PRL-secreting, and 5 ACTH-secreting pituitary adenomas from patients
who had undergone hyphophysectomy at So Jos Hospital of Irmandade Santa Casa de Misericrdia in Porto Alegre, were
included in this study. RNA was obtained by Trizol method and cDNA was synthesized using Superscript 1st strand (Invitrogen).
We evaluated the tumor-type specific MEG3 and GADD45 gene expression by quantitative RT-PCR and correlated with clinical
features. The qRT-PCR was performed using 10L working mix containing 2L of the cDNA template in 2x TaqMan
universal Master Mix (Applied Biosystems) and 100nM final concentration of the primers and the probe. A comercial pool of
RNA from normal pituitary gland was used as a calibrator. The quality control was done by the simultaneous amplification of the
constitutive gene GAPDH. Statistical analysis was done using Kruskal-Wallis followed by post-hoc evaluation. To compare
median between groups, (clinically non-functioning adenomas vs. functioning adenomas) Mann-Whitney test was performed.
Significance was taken at the 5% level. The presence of the GAPDH amplicon by duplicate qRT-PCR confirmed individual
cDNA integrity in all of the samples examined. MEG3 expression was lost in 20 of 38 (53%) pituitary adenomas. Thus far,
clinically non-functioning pituitary adenomas had a significant decrease in MEG3 relative expression compared with functioning
adenomas (p=0,001). In contrast, qRT-PCR showed detectable and variable relative expression of this gene in all functioning
pituitary adenomas (p=0,001), while GH-secreting adenomas (p=0,01) also had a significant variability when compared with
clinically nonfunctioning adenomas. In the same background, GADD45 transcript was not detected in 24 of 38 adenomas
(63,2%). Interestingly, clinically nonfunctioning adenomas had a significant decrease (pGADD45 relative expression when
compared with functioning adenomas. However, some functioning pituitary adenomas demonstrated detectable and significant
variable GADD45 relative expression (pp=0,001) presented increased and variable levels of GADD45 expression when
compared with other tumors, followed by ACTH-secreting adenomas (p=0,05).
Conclusions:
MEG3 and GADD45 gene expressions are significantly lost in almost all clinically nonfunctioning adenomas studied, which are
of gonadotroph origin. Other pituitary tumor phenotypes examined expressed both genes in significantly different levels, and
even with overexpression in some cases. These data suggest that those genes probably were in association with uncontrolled cell
growth and tumor development in the human pituitary gland.
Keywords: Cell Cicle, GADD45, MEG3, Pituitary Adenomas
Financial Support: Programa de Apoio Pesquisa (PROAP) - UFCSPA
Resumo:13-086
VITAMIN D RECEPTOR GENE POLYMORPHISMS AND SEX STEROID SECRETION IN GIRLS WITH
PRECOCIOUS PUBARCHE IN SOUTHERN BRAZIL.
Santos, B. R. 1,2; Mascarenhas, L. P. G. 3; Satler, F. 2; Boguszewski, M. C. S. 3; Spritzer, P. M. 1,2

1
Department of Physiology, UFRGS
2
Division of Endocrinology/HCPA, UFRGS
3
Department of Pediatrics, UFPR
Objectives:
Evidence suggests that precocious pubarche [PP] girls may have higher risk of developing polycystic ovary syndrome [PCOS] at
later ages. Vitamin D receptor [VDR] gene polymorphisms have been implicated in the risk of diabetes and PCOS, but little is
known about the role of VDR in PP. The aims this work were to assess the frequencies of VDR gene ApaI, TaqI, BsmI and FokI
polymorphisms and to determine whether these variants are associated with sex hormone concentrations in patients with PP and
controls from southern Brazil.
Blood was collected from 36 girls with PP (age 11.27 3.79 years at the time of the study) and 197 controls (12.92 2.09 years
at the time of the study) for genotyping of BsmI and FokI polymorphisms using real time polymerase chain reaction [PCR] and of
ApaI e TaqI polymorphisms using restriction fragment length polymorphism [PCR-RFLP], all in vitamin D receptor gene.
Hormone levels were also determined. Genotype GG of the ApaI single nucleotide polymorphism [SNP] was more frequent in PP
(30.6%) than in controls (16.2%) (odds ratio [OR]: 2.269; confidence interval 95%[95%CI]:1.015-5.076; p = 0.042). This
genotype was also associated with lower estradiol (35.30 [14.80-50.48] pg/mL vs. 12.22 [6.49-23.69] pg/mL; p = 0.030) and total
testosterone levels (0.52 [0.39-0.84] ng/mL vs. 0.20 [0.11-0.47] ng/mL; p = 0.009), but not DHEAS (148.20 [79.25 - 202.50]
g/dL vs. 83.03 [62.00 - 174.00] g/dL; p = 0.186), as compared with the TT + TG genotypes in girls with PP. The distribution
of TaqI, BsmI and Fokl SNPs was similar in PP and controls, and no association was found between these polymorphisms and
sex steroid levels.
Conclusions:
The present findings suggest that the ApaI polymorphism of the VDR gene is associated with PP and seems to modulate ovarian
steroid secretion in affected girls. Further studies are needed to better clarify the biological mechanisms by which the ApaI
polymorphism influences PP risk and sex steroid secretion.
Keywords: vitamin D receptor, precocious pubarche, steroid hormones, single nucleotide polymorphism
Financial Support: National Institute of Hormones and Womens Health
Resumo:13-087
MODULATION OF PI3K/AKT PATHWAY BY RENIN-ANGIOTENSIN SYSTEM INHIBITION IN OBESITY
EXPERIMENTAL MODEL.
Bulla, M. L. ; Bertholdo, M. C. D. M. ; Haidar, A. A. ; Hirata, A. E.

Fisiologia/Universidade Federal de So Paulo , UNIFESP
Objectives:
The objective was to evaluate IRSs/PI3-K pathway in insulin resistant obese animals treated with AT1 blocker.
Insulin resitant animals were obtained by neonatally treatment with glutamate monosodium (MSG). MSG animals had higher
body weight than the control animals at seven months old (7-mo). Furthermore, 7-mo MSG animals showed no change in
pressure blood compared to controls and treatment with losartan did not alter it. Evaluating the association of IRS1 with PI3K in
muscle, we noted that the acute stimulation with insulin can promote the association in the control group that received vehicle.
Muscle of MSG animals presented decreased IRS1/PI3K interaction after acute insulin stimulation however no statistical
significance. The treatment with losartan did not change this response. However, IRS2/PI3-K association increased significantly
after losartan treatement. When evaluating the adipose tissue of these animals, we observed an improvement not statistically in
the IRS1/PI3K interaction comparable to the controls suggesting higher sensitivity of this tissue to the hormone But with that
increase group experiemntal. On the other hand, when assessing IRS2/PI3K interaction we observed a strong tendency decrease
in adipose tissue of MSG suggesting locally resistance to insulin action.
Conclusions:
Neonatal treatment with monosodium glutamate (MSG) induced metabolic syndrome. Treatment with losartan did not alter
parameters such as body weight and blood pressure of animals MSG. Treatment with losartan for one week showed a tissue
specific modulation of IRSs/PI3K interaction. Possibly the increased insulin resistance reflected by lower IRS2/PI3K association
in adipose tissue after losartan treatment may contribute to decreased glucose uptake in this tissue and thus would tend to
decrease fat mass and production of various adipokines that are known to work for the installation and worsening of metabolic
syndrome. Our results are not conclusive, being necessary to perform experiments complemetares increasing n order to elucidate
results.
Keywords: Diabetes, Hypertension, Insulin
Financial Support: Scientific activity performed from January 2010 to 2011 grant from FAPESP.
Resumo:13-088
IDENTIFICATION OF NOVEL VARIANTS OF RET ONCOGENE POTENTIALLY LINKED TO THE
PATHOGENESIS OF PHEOCHROMOCYTOMA AND MEDULLARY THYROID CARCINOMA
Bim, L. V. ; Cerutti, J. M.
Universidade Federal de So Paulo, UNIFESP
Objectives:
The RET gene encodes a transmembrane receptor with tyrosine kinase activity linked to signaling pathways involving growth,
migration and cell development. Activating mutations of RET in cell germlines are associated with an autosomal dominant
hereditary syndrome called multiple endocrine neoplasia type 2 (MEN 2), which can be classified into three subtypes: familial
medullary thyroid carcinoma (FMTC), MEN 2A and MEN 2B. MEN 2A is characterized by the presence of medullary thyroid
carcinoma (MTC), pheochromocytoma (PHEO) and hyperparathyroidism. FMTC is characterized by the presence of MTC as the
only clinical feature. Our group recently described a novel heterozygous mutation in exon 8 of RET gene associated with FMTC,
which leads to the substitution of a glycine by a cysteine at codon 533 (p.G533C). Later a member of the family with FMTC was
diagnosed with PHEO, suggesting that p.G533C mutation leads to the MEN 2A phenotype. Interestingly, in addition to the RET
mutation p.G533C, two novel variants within RET oncogene were found in DNA isolated from PHEO, which were not detected
in the DNA extracted from the blood of the patient. This finding suggests that additional genetic events in somatic tissues may be
associated with tumorigenesis of MEN 2A-associated tumors. Whether this event is associated with pathogenesis of sporadic
PHEO or MTC, is still unclear. We here aim to investigate the prevalence of new variants in the RET gene exon 8 (G548V) and
exon 9 (S556T) in sporadic and familial PHEO and MTC. We additionally will investigate whether these variants activate the
MAPK pathway, through in vitro phosphorylation of ERK.
A set of 25 PHEO and 43 MTC, sporadic and MEN 2A-associated tumors were selected from the files of the Department of
Pathology of UNIFESP. Detection of RET variants in paraffin-embedded pheochromocytoma specimens will be performed by
PCR sequencing. Specific primers were designed for amplification of exons 8 and 9 of the RET gene. Of the 12 samples, 6 have
already had the genetic material extracted and primers are in the process of standardization. For functional analysis, cDNA RET
Wt and new mutants cDNA will be cloned in expression vector (pBUD4.1), there will be a site-directed mutation to obtain the
G533C mutation, the plasmid will be then transfected into HEK 293 cells for protein expression and RET phosphorylated ERK
analysis by Western blotting. Specific primers for subcloning and site directed mutation of RET were designed and are being
standardized.
Conclusions:
Recent data on the exon 8 mutations and its genotype-phenotype suggest that they are more aggressive and less rare than
originally imagined. The two new mutations never described before, found in the patient, suggest that a second hit in the RET
gene can be a potential mechanism for the formation of pheochromocytoma in patients with MEN 2A. Thus, the results of this
project can and aims to help clarify such issues.
Keywords: MTC, Mutations, Pheochromocytoma, RET oncogene
Resumo:13-089
INTERMITTENT APNEA INDUCES INSULIN RESISTANCE IN CONSCIOUS RATS.
Quadros, C. D. M. 1; Haidar, A. A. 1; Nejm, M. B. 3; Rocha, M. S. 2; Carpinelli, A. R. 2; Cravo, S. L. D. 1;

Schoorlemmer, G. H. M. 1; Hirata, A. E. 1
1
Depto. de Fisiologia/ Universidade Federal de So Paulo, UNIFESP
2
Depto. de Fisiologia e Biofsica, Universidade de So Paulo, USP
3
Depto. de Neurocincias/ Universidade Federal de So Paulo, UNIFESP
Objectives:
Diabetes mellitus is a very common finding in patients with sleep apnea, but whether apnea causes diabetes, diabetes causes
apnea, or both are consequences of obesity is not clear. We used a new method to induce intermittent apnea in rats and measured
the consequences of apnea on glucose tolerance and insulin sensitivity.
We used male Wistar rats weighing 300 500 g. Rats were anesthetized with ketamine and xylazine, and a balloon-tipped
polyurethane catheter was implanted in the trachea. The balloon was contained in a rigid Teflon tube so that inflation of the
balloon blocked the airway, without inducing tracheal pain. A silicone rubber cannula was inserted in the femoral vein and
advanced to the thoracic vena cava. Both balloons were exteriorized on the rats back. These implants allow induction of apnea
and removal of blood samples in conscious, unrestrained rats. Rats were allowed at least a week to recover pre-operative body
weight. To measure insulin sensitivity, insulin (0.75 U/kg body weight) was injected through the venous cannula. Blood glucose
concentration was measured just before, and 4, 8, 12, and 16 min after insulin injection. To measure glucose tolerance, glucose (1
g/kg body weight) was injected through the venous cannula, and blood glucose concentration was measured just before, and 4, 8,
12, 16, 20, 24, and 28 min after glucose injection. Glucose tolerance and insulin sensitivity were measured a few days before and
30 min after 8 h of intermittent apnea (14 s every 2 min). Apnea did not significantly change glucose tolerance (AUC in controls
1110 52 mg/dl*min, after apnea 1181 55 mg/dl*min, n = 8/5), but it significantly reduced insulin sensitivity: the glucose
decay constant fell from 5.3 0.4 to 3.9 0.5 %/min (n=8/7).
Conclusions:
Our data suggest that apnea contributes to development of insulin resistance. However, we are not yet sure that the mechanism of
insulin resistance is the same in human patients with sleep apneas and in rats with a tracheal balloon. We are planning to increase
the delay between the end of the apneas and measurement of insulin sensitivity to reduce acute effects of stress.
Keywords: Glucose tolerance, Insulin resistance, Intermittent apnea
Financial Support: CAPES e FAPESP
Resumo:13-090
THYROID STATUS MODULATES THE EXPRESSION OF THYROID HORMONES TRANSPORTERS MCT8 AND
MCT10 IN THE LIVER
Pereira, G. F. ; Ramos, R. G. ; Imprio, G. E. D. ; Santiago, L. A. ; Faustino, L. C. ; Ortiga-carvalho, T. M.

Objectives:
The idea that thyroid hormones (TH) entered cells passively through the plasma membrane was abolished after the discovery of
transporters that mediate the cellular influx and efflux of TH. Two members of the monocarboxylate transporters (Mct) family
can transport TH, Mct8 and Mct10, which are expressed in many tissues. However, it is unknown whether TH can change its
transport into liver cells via regulation of its transporters. Here, we aim to evaluate the mRNA expression of Mct8 and Mct10 in
the mice liver after TH deprivation, chronic treatments with both TH biologically active forms (T3 and T4) and acute treatment
with T3 only.
In 12-week-old male mice, hypothyroidism (Hypo group) was induced by feeding them a 0.15 % PTU diet for 3 weeks. Daily sc
injections of T3 (50 g /100g BW) were given for 2 weeks to induce hyperthyroidism in the group called HyperT3, while sc
injections of T4 at the same concentration were given during the same period to induce hyperthyroidism in the group called
HyperT4. Four animals were used in each group, except the control group, composed by five animals, totalizing 17. Acute
treatment was done by one single sc injection of T3 (50 g/100g BW). Mice were sacrificed 15, 30, 60, 180 and 360 minutes
after injection. Control groups were injected with saline. Data were reported as mean SEM. One-way ANOVA followed by
Student-Newman-Keuls multiple comparisons test and Two-way Anova were employed. Five animals were used in each group,
resulting in 50 treated animals. Liver mRNA levels of Mct8 and Mct10 were analyzed by real time PCR, samples were
normalized to euthyroid controls. Unexpectedly, in chronically treated mice, both hypo- and T3 treated hyperthyroidism led to the
reduction of mRNA expression of Mct8 (Hypo: 0.5 0.01; HyperT3:0.40.1,p0.05). However, there is a tendency to increase the
mRNA expression in HyperT4 compared to HyperT3, although not significantly different. Conversely, acute T3 injection
increased mRNA expression of Mct8 (60:2.30.2,180:1.60.14,360:1.90.37) p

Conclusions:
Our preliminary data showed that, chronically, hypothyroidism and T3 treatment decreased Mct8 and Mct10 mRNA expression
in mice liver. Acutely, T3 increased Mct8 and Mct10 mRNA expression, suggesting a non genomic action of T3.
Keywords: thyroid hormones transporters, thyroid status, liver
Financial Support: CNPq, CAPES, FAPERJ, Dept. Tireide SBEM.
Resumo:13-091
ADHERENCE OF PATIENTS IN AN EDUCATION AND CONTROL PROGRAM OF DIABETES IN THE CITY OF
JATA - GO
Andrade, D. O 1; Ferreira, N. S. 1,1; de Lira, C. A. B. 1,1,3; Ferri, L. P. 4; Moraes, L. C. 4; Cintra, C. E. 4;

Benite-ribeiro, S. A1
1
Universidade Federal de Gois, UFG
2
3
4
Secretaria Municipal de Sade, SMS
5
Secretaria Municipal de Sade, SMS
6
Objectives:
Diabetes is an important metabolic disease and it is a foremost public health problem, affecting more than five million Brazilian
individuals. The patient with diabetes can be benefit from intervention programs that focus on drug treatment and nonpharmacologic procedures, such as regular physical exercise (PE), diet and body mass (BM) control. So, adherence (AD) to
therapy is essential to success of treatment. AD can be defined as the extent to which a person performs recommendations from a
health care provider. In Jata GO, there is a Program of Education and Control of Diabetes (PECD), conducted by a
multiprofessional health care team (MHC). In this program, patients are given drug therapy and are advised to practice PE and
hypocaloric calorie diet (HCD). Thus, this study aimed to assess patients AD to the PECD and the effects of counseling done by
the MHC.
The study included patients with diabetes treated by the PECD. We evaluated the number of patients who after being registered in
the program (1st visit, n=102) attended all the queries until the 4th and 8th visits. We analyzed the prevalence of patients with
postprandial plasma glucose (PG) decompensated (PG 180 mg/dL), patients with overweight (BMI 25m.cm2) and the patients
reports about the practice of PE and HCD. The prevalence were tested by Chi square test and the statistical significance was
p0.05. Data are presented as meanSD. It was observed that of 102 patients registered at the 1st consultation only 44 (43.1%)
remained until 4th visit, and 20 (19.6%) patients continued until 8th visit, showing a low frequency of patients to the queries. At
the 1st visit, there was prevalence of overweight (75.8%), and the percentage of patients with decompensated PG was 59.3%,
non-practicing HCD was 43.1%, and non-practicing PE was 46%. At 4th visit there was prevalence of overweight (71.9%) and
practicing PE (70.4%), where as 48.3% have not yet followed HCD. Although there are 20 patients in the 8th consultation, the
records were incomplete, and were not possible to do statistical analysis. Despite of weak AD, patients who did both the PE and
HCD had compensated PG at 4th consultation (in mg/dL): PE practicing= 15932.71, PE non-practicing= 182.988.78; HCD
practicing= 149.958.78, and HCD non-practicing= 202.3102.0.

Conclusions:
In this study was observed low AD of the patients to the PECD, both by reducing the number of patients who attended all visits,
as by no patients AD to advice from the MHC. Problems with AD to therapeutic recommendations are common in almost all
diseases, which impact negatively the effectiveness of the treatment. Probably the factors related to the AD are due to the
complexity of the therapeutic schedule and the adaptability of the recommendations to the usual habits of the person. Indeed, the
knowledge of the patients about disease, the relationship with the health care time and the patients perception of health and the
benefits of treatment are factors associated with adherence. So, we suggest that the interventions need to be easier to apply and
that the MHC combined cognitive and behavioral strategies, planning activities of community meetings to increase patients
knowledge about the disease and the benefits of AD to the program. Nevertheless, patients who adhered to the PECD had better
control of PG, demonstrating the importance of MHC work in the control of diabetes.
Keywords: diabetes, adherence, physical exercise, hypocaloric diet, multiprofessional health care
Resumo:13-092
SERUM AMYLOID A (SAA) EXPRESSION IS MODULATED BY HYPOXIA IN ADIPOCYTES
Oliveira, E. M. ; Monteiro, F. B. F. ; Sandri, S. ; Knebel, F. H. ; Contesini, C. G. I. ; Campa, A.

Department of Clinical Chemistry / FCF, USP
Objectives:
In obesity, adipocyte hypertrophy and hyperplasia might generate hypoxic areas within the tissue. In a close relationship with
hypoxia, the adipose tissue produces inflammatory molecules involved in obesity-related complications, such as cardiovascular
disease and metabolic disorders. Besides the hypoxia-inducible factor 1 (HIF-1), several proteins have their stability or
expression modulated by hypoxia. It was also recognized that adipocyte size was correlated with the expression and production of
the serum amyloid A (SAA). This protein acts as a potent stimulus for the production of cytokines in immune cells. Thus, the aim
of the present study was to assess the influence of hypoxia on the expression of adipocyte-derived protein SAA isoforms, and
investigate, in normoxia, the effect of SAA on the production of proinflammatory cytokines by 3T3-L1 cells and human
adipocytes.
The cell culture was performed according to a standard protocol using a murine fibroblast cells (lineage 3T3-L1) from ATCC and
human adipocytes from abdominal subcutaneous adipose tissue, derived from plastic surgery and obtained from five healthy
donors (BMI between 25 and 35). The ambient hypoxia was generated by filling in a sealed acrylic chamber (self-designed) with
low-oxygen air that contained 1% oxygen, 5% carbon dioxide, and 94% nitrogen. Western blotting was used to measure HIF-1
and SAA; for the last, Real Time PCR analyses were also performed. The production of the cytokines TNF-, IL-6, IL-1 and IL8 in cell culture supernatants was determined by ELISA assays. Ambient hypoxia (1% O2) caused a two (2,02 0,08 *p
Conclusions:
Our data show that the SAA gene is responsive to hypoxia, and that SAA possibly favors the improvement of the inflammatory
profile of the adipose tissue that may contribute to the overall health complications related to obesity, including insulin resistence.
The study was approved by the Faculty of Pharmaceutical Sciences Human Ethics Committee.
Keywords: 3T3-L1, CYTOKINES, HYPOXIA, HUMAN ADIPOCYTE, SERUM AMYLOID A
Financial Support: FAPESP; CAPES; CNPq.
Resumo:13-093
THIRST AND SODIUM APPETITE: EFFECT OF ADRENALECTOMY AND ESTRADIOL TREATMENT IN
OVARIECTOMIZED RATS
Almeida-pereira, G. ; Elias, L. L. K. ; Antunes-rodrigues, J

Physiology Dept/ FMRP-USP, USP
Objectives:
The regulation of body fluids in many species is under the influence of ovarian hormones, especially estrogen. In addition, there
are several experimental evidences demonstrating the involvement of hypothalamic pituitary adrenal axis (HPA) in the control of
hydromineral homeostasis, as well as the influence of estrogen in the modulation of this axis. In this context, our objective was to
study the influence of estrogen replacement in ovariectomized rats on the activity of the HPA axis using the adrenalectomized
animal, with or without hormone replacement, for the evaluation of behavioral responses involved with control of body fluids
homeostasis.
Wistar rats (220-240g, N = 11-14) were bilaterally ovariectomized and treated with estradiol (20 g/animal/day, sc, OVX-E2) or
vehicle (corn oil, 0.2 ml daily, sc, OVX) started 24 after surgery and maintained for 14 days. On the seventh day of replacement
the animals were submitted to bilateral adrenalectomy (ADX-OVX, OVX-E2-ADX) or sham surgery (SHAM-OVX, OVX-E2SHAM) and they were treated with corticosterone (10 mg/kg/day, sc, OVX-SHAM-B, OVX-E2-SHAM-B, OVX-ADX-B, OVXADX-B-E2) or vehicle (corn oil containing 5% ethanol, sc, 0.2 ml daily) for 7 days. Animals pre-habituated to the metabolic cage
had the daily water and hypertonic saline (1.8%) intake evaluated (18:00h) during the six days after ADX or sham surgery. The
results are expressed as mean SEM of cumulative intake. Data analysis were performed using three-way ANOVA, followed by
post-test Newman-Keuls and the level of significance was set at p<0.001).
Conclusions:
Our results demonstrate that E2 attenuates sodium intake induced by ADX, can be postulated that E2 might reduce ANGII
secretion, and thereby reducing sodium intake after ADX. However, the effect of E2 reducing sodium appetite could involve
regulation of other factors than ANGII, since this effect was also observed in animals with intact adrenals treated with B. The E2
also modulates the mechanisms involved in water intake in ADX animals, but only in the presence of B, since in group OVX-E2ADX-B was observed reversal of the effect induced by ADX.
Keywords: Adrenalectomy, Corticosterone, Estrogen, Sodium intake
Financial Support: CAPES, CNPq and FAPESP
Resumo:13-094
MODULATION OF GLUCOSE TRANSPORTERS GLUT2 AND SGLT2 IN KIDNEY: PARTICIPATION OF AKT,
INSULIN AND HNF-3 BETA TRANSCRICIONAL FACTOR.
Freitas, H. S. ; Okamoto, M. M. ; Beloto - Silva, O. ; Sabino-silva, R. ; David-silva ; Furuya, D. T. ; Souza,

M. O. ; Machado, U. F.
Dep. Fisiologia e Biofsica - Inst. de Cincias Biomdicas , ICB / USP
Objectives:
The Akt is one downstream target of phosphatidylinositol 3-kinase and plays an important role in mediating effects of insulin on
hepatic glucose production, glycogen, and protein synthesis. Upon activation, Akt is translocated to the nucleus where it exerts
effects on gene activity by phosphorylation of target proteins. Genetic studies have shown that the activation of PI3-kinase-Akt
pathway by insulin induces HNF-3 phosphorylation; this transcricional factor physically interacts with Akt and is phosphorylated
at a single site (T156). Renal glucose reabsortion is a coordinated process, which takes place in the epithelial cells of the proximal
tubule, involving two classes of glucose transporters, the Na - glucose transporters (SGLTs) and the facilitative diffusion
transporters (GLUTs). Increases in the renal glucose transporter gene expression are involved in renal tubule - glomerular
diseases. We showed that GLUT2 and SGLT2 are increased in diabetes and treatment with insulin (4 hours to 2 days) in kidney
of rats. That overexpression of GLUT2 and SGLT2 is related to the increased expression and activity of HNF-3 beta transcription
factor. Thus, the aim of this study is to establish involvement of the insulin, kinase Akt and HNF-3 beta in GLUT2 and SGLT2
modulation in renal territory.
The GLUT2, SGLT2, HNF-3 and p-Akt (Ser473) expressions as well the activity of HNF-3 transcription factor were analyzed in
IRPTC (immortalized renal proximal tubular cells) cells cultivated under such conditions: low (L) or high glucose (H), high
glucose plus insulin (I), high glucose, insulin and Akt inhibitor (Ii), and high glucose plus Akt inhibitor (Hi). In comparison to L,
p-Akt expression was increased in H (498.015, P<0.001).
Conclusions:
Our results clearly demonstrated that insulin regulates GLUT2 and SGLT2 expressions by increasing p-Akt protein, HNF-3 beta
expression and its binding activity. Moreover, this study suggests that Akt could play a key role in renal glucose reabsorption, by
modulating glucose transporters expressions.
Keywords: Rim, GLUT2, SGLT2, Insulina, Akt
Financial Support: FAPESP 05/60588-5 and 07/50554-1
Resumo:13-095
PROGRAMMING OF ADRENAL MEDULLARY FUNCTION BY NEONATAL OVERFEEDING IN RATS
Santos, A. S. ; Conceio, E. P. S. ; Oliveira, E. ; Pinheiro, C. R. ; Trevenzoli, I. H. ; Cardoso, F. S. ;

Moura, E. G. ; Lisboa, P. C.
DCF/UERJ, IBRAG
Objectives:
Several studies reveal that nutritional, hormonal and environmental factors at prenatal and neonatal period may influence organs
and tissues development, and this could be related with late diseases, like diabetes and cardiovascular disturbances (1, 2).
Postnatal overfeeding increases development risk to obesity and cardiovascular diseases. It has been shown that overfed neonate
rats show higher visceral adiposity and hyperleptinemia at weaning and they were programmed for obesity, higher food intake
and hypertension at adulthood (3). Previously, we evidenced that neonatal hyperleptinemia induces adrenal medullary
hyperfunction in early and late life (4). In the present study, we evaluated the adrenal function of obese adult rats that were
overfed during lactation. We also evaluated indirectly the visceral adipocyte sensitivity to serum catecholamine, through 3adrenergic receptor (ADRB3).
To induce early obesity by overfeeding, the litter size was reduced from ten to three male pups at third day of lactation until
weaning (small litter group, SL) while the control group stayed with ten males by litter over the lactation (normal litter group,
NL). After weaning, one pup from each litter (8/group) had free access to standard diet and water until the 180 days old when
they were killed and were collected samples of visceral adipose tissue (VAT), liver and the adrenal glands. Significant differences
had p
Conclusions:
The higher adrenergic catecholamine profile can help to explain the reported hypertension in this model. Paradoxically, those
animals had higher liver glycogen content and VAT, which suggest a participation of other factors than adrenaline, such as a
preferential use of lipids as energy source, saving glycogen.
Keywords: adrenal medullar function , catecholamine, postnatal overfeeding
Financial Support: CNPq, FAPERJ and CAPES
Resumo:13-096
AMP-ACTIVATED PROTEIN KINASE MEDIATES GLUCOSE UPTAKE IN RAT THYROID CELLS THROUGH A
TSH-INDEPENDENT MECHANISM
Cazarin, J. M. 1; Andrade, B. M. . 1; Rodrigues, D. C. 1; Zancan, P. 2; Ceddia, R. B. 3; Carvalho, D. P. 1

1
Instituto de Biofsica Carlos Chagas Filho - UFRJ, IBCCF/UFRJ
2
Faculdade de Farmcia-UFRJ, FF/UFRJ
3
School of Kinesiology and Health Science - Faculty of Health, York University
Objectives:
Glucose is an essential substrate for several cell functions. Glucose uptake is mediated by a family of glucose transport proteins
(GLUTs) expressed in a tissue-specific manner. Several glucose transporters are expressed in rat thyroid lineage cells and rat
thyroid gland with predominant expression of GLUT-1. However, the mechanisms that regulate GLUT1 expression and
translocation in thyroid cells are not completely understood. Several studies described that some thyroid carcinomas exhibit a
phenotype of low iodide uptake and increased glucose uptake. Recently our group showed for the first time that AMP-activated
protein kinase (AMPK) is expressed in the rat thyroid gland and decreased iodide uptake by the thyroid follicular cell, modulating
a major step involved in thyroid function (Am J. Physiol, in press, March 2011). Although many studies demonstrated that
AMPK regulates glucose uptake in different tissues, none have looked to this potential role of AMPK in glucose metabolism in
thyroid cells. Therefore, the study of AMPK signaling effects in thyroid cell can help to understand some acpects of the thyroid
carcinogenesis. The purpose of this study was to investigate the role of AMPK in the regulation of glucose uptake in rat thyroid
cells.
PCCL3 cells, a thyroid cell lineage, were cultivated in Ham's F12 medium supplemented with 5% fetal bovine serum and
containing six components: 1mU/ml bovine TSH, 10g/ml insulin, 5 g/ml transferring, 10nM hydrocortisone, 6nM
Somatostatin, and 2.5M Gly-His-Lys. The cells were then treated with an AMPK activator, AICAR (A-1mM), or an AMPK
inhibitor Compound C (CC-20M), or both. Besides, some cells were submitted to TSH deprivation for 24h. The incubation with
AICAR for 24h induced a significant (C 1,01&plusmn0.05; A0.5mM 0.87&plusmn0.02; A1mM 1.55&plusmn0.13; A2mM
2.00&plusmn0.05; n=6 P<0.05).
Conclusions:
Our results suggest that AMPK regulates glucose uptake in rat thyroid cells independent of the TSH, and might be involved in the
increased glucose uptake that is characteristic of tumoral cells.
Keywords: AMPK, Thyroid, Glucose uptake, GLUT-1, PCCL3
Financial Support: FAPERJ, CNPq and CAPES
Resumo:13-097
IN VIVO CROSS-TALK BETWEEN THYROID HORMONES AND ESTRADIOL IN RENAL GLUTATHIONE STRANSFERASE ALPHA REGULATION
Faustino, L. C. 1; Almeida, N. A. 2,1; Pereira, G. F. 1; Ramos, R. G. 1; Santiago, L. A. 1; Cordeiro, A. 1;

Ortiga-carvalho, T. M. 1
1
IBCCF, UFRJ
2
Depto de Ciencias Fisiolgicas, UFRRJ
Objectives:
Aim: Glutathione S-transferase alpha (Gsta) represents an essential component of cellular antioxidant defence mechanism in
liver, and also in kidney. Mice liver Gsta mRNA was described to be repressed by thyroid hormones (TH) only in hypothyroid
state. Our previous data have shown an essential role of TRbeta1 in mediating liver Gsta suppression in response to T3, and in the
absence of a functional TRbeta, there is a compensatory action of TRalpha1. Kidney is one of the most important tissues
responsible for detofication processes. Until today, there is no published data about Gsta regulation by TH in the kidney.
Recently, we observed gender-dependent kidney Gsta regulation. In males, we saw an increase of Gsta levels in hypothyroidism
whereas in females, surprisingly, there was an unexpected reduction of renal Gsta expression suggesting a possible cross talk
between TH and estradiol/progesterone regulation pathways. Therefore, this work aims to evaluate, in vivo, the interaction among
TH and sex hormones in kidney Gsta expression.
All animals used in this work were 8-week-old females subjected to ovariectomy (OVX). After OVX, mice were induced to
hypothyroidism (HYPO) and hyperthyroidism (HYPER). HYPO was induced by PTU diet for 4 weeks. Mice were rendered
hyperthyroid by daily sc injections of T3 (50 g/100 g BW) for 15 days. Aiming to investigate which ovarian sex hormone could
be responsible for gender differences in renal Gsta expression regulation, we performed OVX followed by treatment with
estradiol benzoate (Es). Mice received Es injections at 0.1 mg/kg BW for 15 days. One control group was sham operated females
and another control group was male mice. In another experiment, after OVX and TH modulation, mice were also treated with Es
for 15 days. Gsta mRNA and protein levels were evaluated using quantitative RT-PCR and Western Blot, respectively. After
ovariectomy (OVX), Gsta mRNA was increased in HYPO (2.20.2; p
Conclusions:
Our data show a synergic interaction of TH and estradiol regulating renal Gsta expression, in which Gsta appears to be
downregulated by T3 in the absence of estradiol, and upregulated by T3 when estradiol is present.
Keywords: thyroid hormones, estradiol, glutatione S transferase, kidney, cross-talk
Financial Support: CNPq, CAPES, FAPERJ, Dept. Tireide SBEM.
Resumo:13-098
MORPHOLOGICAL ANALYSIS OF FIBER-SKELETAL MUSCLE OF ADULT RATS SUBMITTED TO
PERINATAL PROTEIN MALNUTRITION
Tfolo, L. P. ; Mendes, F. C. V. ; Ribeiro, T. A. D. S. ; Fabricio, G. S. ; Oliveira, J. C. D. ; Torrezan, R. ;

Agostinho, A. R. ; Silva, P. M. D. S. ; Lopes, D. A. ; Rinaldi, W.
Depart of cell biology and genetic, UEM
Objectives:
It has been showed that early undernutrition is associated with being overweight in adult life and leads serious pathophysiologys
such as cardiovascular diseases, hypertension and type 2 diabetes. The low protein diet during early life might lead to irreversible
changes on glucose homeostasis, insulin secretion and hypoinsulinemia in rats. Insulin is an important hormone for growth and
development of the muscle. Thus, the objective this work was to study the effects of metabolic programming by maternal protein
restriction in the morphology of skeletal muscle.
Dams received a poor protein diet (4%) during initial 2/3 of lactation (RP), while, control-dams (NP) received normal protein diet
(23%), after that both offspring ate normal diet until 90-day-old. Adult rats were sacrificed and biometric parameters were
analyzed. The retroperitoneal and periepididymal fat pads were removed and weighed. The soleus muscle was removed, weighed
and frozen in liquid nitrogen. The cross sections area soleus muscle (120 fibers/animal, 10 fibers/field, 3 fields/cut and 4
cut/animal of each experimental group) were staining by hematoxylin and eosin (HE), after that the images were taking through
the stereomicroscope to subsequent quantification. Results were evaluated through the Students t-test (p
Conclusions:
Maternal protein restriction during lactation does not impair the morphology of the soleus muscle.
Keywords: Protein restriction , insulin secretion, soleus muscle, glucose homeostasis
Financial Support: CNPq, CAPES and Fundao Araucria
Resumo:13-099
INTERFERON-GAMMA DECREASES RAT PINEAL NFKB NUCLEAR TRANSLOCATION AND POTETIANTES
MELATONIN PRODUCTION
Barbosa Lima, L. E. ; Fernandes, P. A. C. M. ; Markus, R. P.

Dep. de Fisiologia/ Instituto de Biocincias-USP, IB-USP
Objectives:
The cross-talk between the pineal gland and the immune-system characterizes an immune pineal axis (Neuroimmunomodulation
14, 126, 2007). We have previously shown that the activation of nuclear factor kappa B (NFKB) by TNF and LPS pathway
inhibits melatonin production, while inhibition of NFKB by glucocorticoids has the contrary effect. In order to understand how
the pineal gland activity is altered during the development of an inflammatory response, we aim to evaluating the effect of other
cytokines that peaks lately than TNF. In the present work, we analyzed the effect of interferon (IFN)-gamma, by determining
NFKB nuclear accumulation and noradrenaline-induced melatonin synthesis.
Male Wistar rats aging 2-3 months were used in all the experiments. All animal procedures were performed according to
approved institutional protocols (048/2007). Pineal glands cultivated in BGJB medium for 48 h were incubated with IFN-gamma
(30 ng/mL) for 1, 5, 10, 15 or 30 min. In order to determine the effect of IFN-gamma on noradrenaline-induced melatonin
production, the glands were pre-incubated for 30 min with IFN-gamma (10 or 30 ng/mL) and then noradrenaline (10 nM) was
added for 5 h. NFKB nuclear content was determined by electromobility shift assay (EMSA), and melatonin content in the
medium was measured by HPLC. Maximal inhibition of NFKB nuclear translocation by IFN-gamma (30 ng/mL) was observed
after 10 min (28.1 11.4%, n=3, compared to vehicle). This effect is probably transient, as after 15 min there was an increase of
the percentage of nuclear NFKB (54.0 15.1%, n=3). The same profile of IFN-gamma effect was observed in the glands
incubated IFN-gamma plus noradrenaline. In addition, IFN-gamma (10 and 30 ng/mL) potentiates noradrenaline-induced
melatonin basal production (24.3 4.2 ng/mL, n=8) in 43.6 6.9% (n=4) and 60.4 11.8% (n=4), respectively. Therefore, our
data show that INF-gamma potentiates melatonin, simultaneously with inhibition of nuclear translocation of NFKB.
Conclusions:
Taking into account that the potentiation of melatonin production by glucocorticoids was due to NFKB inhibition, our data
strongly suggest that the effect of IFN-gamma is also mediated by this transcription factor. In addition, we may propose that
during the development of the inflammatory response melatonin pineal production is restored by the interference of IFN-gamma.
Keywords: EIXO IMUNE-PINEAL, INTERFERON-GAMMA, MELATONINA, NFKB, PINEAL
Resumo:13-100
CHALCONE ANALOGUE STUDIES ON INSULIN SECRETION AND IN VITRO DISACCHARIDASES ACTIVITY
IN RATS
Pereira, D. F. 1; Tavares, L. C. 2; Kappel, V. D. 1; Cazarolli, L. H. 3; Guesser, S. 1; Pizzolatti, M. G. 2; Silva,

F. R. M. B. 1
1
Departamento de Bioqumica/Univ. Fed. de Santa Catarina, UFSC
2
Departamento de Qumica/ Univ. Fed. de Santa Catarina, USFC
3
Universidade Federal de Fronteira Sl, UFFS
Objectives:
It is describe that some chalcones show anti-hyperglycemic activity when administered in hyperglycemic normal rats. In
adipocytes, chalcone analogues stimulate the glucose uptake and potentiate insulin-induced glucose uptake. The aim of this study
was to investigate the mechanism of chalcone QC10 on oral glucose tolerance curve improvement, on insulin secretion and in
vitro disaccharidase activity (maltase).
METHODS: Male Wistar rats (180-250 g) were deprived of food for at least 16 h but allowed free access to water. Fasted normal
rats were loaded with glucose (4 g/ kg p.o.) plus different doses of the chalcone (5, 10 and 20 mg/ kg). The glycemia (mg/ dL)
was measured at zero, 15, 30, 60 and 180 min after treatment, by glucose oxidase method. Serum samples from rats untreated
(control) or treated with QC10 (10 mg/ kg), by oral gavage, were used to determine insulin secretion. The insulin levels were
measured by enzyme-linked immunosorbent assay (ELISA) according to the manufacturers instructions. The range of values
detected by this assay was: 0.210 ng/ml. The intra- and inter-assay coefficients of variation (CV) for insulin were 3.22 and 6.95,
respectively, with a sensitivity of 0.2 ng/ml. To disaccharidase activity, duodenum was removed, homogenized in saline and kept
on ice until the moment of the experiment. Aliquots of the homogenate were pre-incubated for 5 min in the absence (controls) or
in the presence of chalcone. After, duodenum homogenates were incubated for 5 min with the substrate maltose. Chalcone was
used in several dosages, such as 25; 50; 100; 200 and 400 mM. The glucose produced was measured by glucose oxidase method
and its concentration was used to determine the maltase activity. Previous data demonstrated the higher activity of maltase in the
duodenum portion (basal activity). RESULTS: Chalcone caused significant acute anti-hyperglycemic effect when compared to
respective hyperglycemic control group (0 min: 114 2; 15 min: 170 3; 30 min: 191 4; 60 min: 170 3 and 180 min: 128 6
mg/ dL). Serum glucose-lowering was detected at 15, 30 and 60 min after oral treatment to any dose tested. The serum glucoselowering was around 12% ate 15 min and 20% at 30 and 60 min for 5 mg/ kg QC10. The anti-hyperglycemic activity was higher
to 10 and 20 mg/ kg QC10 at 15 min: 24%; 30 min: 30% and 60 min: 20%, respectively. The insulin secretion increased at 60
min (1.4 0.2) and can represent the classical second phase of insulin secretion when compared with basal insulin (0.54 0.04).
Chalcone was able to inhibit the specific activity of maltase with 100, 200 and 400 mM when compared with the basal enzyme
activity.
Conclusions:
CONCLUSION: From these results we can conclude that this class of chalcone (QC10) improves glucose tolerance curve,
probably, by acting as insulin secretagogue. Also, an alternative intestinal target to this chalcone (maltase) is associated to
ameliorate glycemia balance.
Keywords: Chalcone, insulin secretion, disaccharidases
Financial Support: CNPq; CAPES, FAPESC, UFSC
Resumo:13-101
CPG ISLANDS SELECTION FOR STUDYNG METHYLATION PATTERNS OF RET (MAPK), HES1 (NOTCH) AND
WNT5A (WNT/-CATENINA) REGULATORY REGIONS IN MEDULLARY THYROID CARCINOMA
Cardoso, M. G. ; Maciel, R. M. D. B. ; Jasiulionis, M. G. ; Ierardi, D. F. ; Aguiar, G. ; Harada, M. Y. ;

Kizys, M. M. L. ; Silva, M. R. D. D.
Objectives:
Differential patterns in DNA 5-methyldeoxycytidine influence gene expression for many genes in development, differentiation,
aging and carcinogenesis. It has been known for decades that Familial Medullary Thyroid Carcinoma (FMTC) is caused by
activating mutation in the oncogene RET, but the role of DNA methylation at its promoter region is scarcely investigated.
Intriguingly, family members bearing the same RET mutation present contrary prognosis, follow-up and tumor progression. This
observation has raised the hypothesis of epigenetics modifying events on oncogenes and tumor suppressor genes as RET
(MAPK), HES1 (NOTCH) and WNT5A (WNT/-CATENINA), genes and cell signaling implicated in carcinogenesis. In this
initial phase of the project we aim at selecting in silico the best predictive RET, HES1 and WNT5A regulatory regions enriched for
CpG islands, thereby, following the study of bisulfite methylcytosine sequence mapping.
We performed a detailed analysis of potential CpG island using the bioinformatics programs: Methprimer and MethylPrimer
Express. We opted to select two regulatory regions for each gene, termed as alternative A and B, hence, to proceed with BSPdesigned primers. We standardized paraffin-tumor DNA extraction, bisulfite treatment and DNA purification for the following
BSP-PCR amplification and then bisulfite genomic sequencing. Bisulfite-treated PCR amplified fragments were also inserted into
pCR4.1 sequencing vector to ensure 16 sequenced colonies for comparison and confirmation for methylated cytosines with direct
PCR product sequence. PCR condition consisted of forty-five cycles at 95C for 1min, 53C for 1min and 72C for 1min using
both primers and final extension at 72C for 10 min. We were able to successfully design BSP primers and bisulfite-sequence
large CpG islands of RET, HES1 and WNT5A regulatory regions, overcoming the challenge of obtaining sufficient amount of
DNA from FMTC paraffin tumor samples, and improve the readability of GC-rich sequences usually seen at the promoter
regions.
Conclusions:
we optimized the conditions for studying methylation pattern changes to understand the discrepancy in inherited tumor phenotype
in patients with same RET mutation. In the second ongoing phase, we will evaluate whether the hypermethylation of HES1 and
WNT5A, and hypomethylation of RET are linked with transcription repression and activation, respectively, therefore, imposing a
worse FMTC prognosis.
Keywords: epigenetics, Thyroid Carcinoma, Methylation, RET, HES1 and WNT5A
Resumo:15-032
IDENTIFICATION OF NOR--LAPACHONE DERIVATIVES AS POTENTIAL ANTIBACTERIAL COMPOUNDS
AGAINST ENTEROCOCCUS FAECALIS CLINICAL STRAIN
Loureno, A. L. 1; Abreu, P. A. 1,2; Rodrigues, C. R. 2; Castro, H. C. 1; Milazzotto, T. L. 1

1
Instituto de Biologia / Universidade Federal Fluminense, UFF
2
Faculdade de Farmcia, UFRJ
Objectives:
A broad-spectrum antibiotic therapy has led to medical complications and emergence of multiresistant bacteria including
Enterococcus faecalis. Naphthoquinones have been focused in a variety of studies because of their various biological
activities. In this study, we designed, synthesized, and evaluated the antibacterial activity of 13 nor--lapachone
derivatives against a drug resistant E. faecalis strain using a molecular modeling approach.
All the substances were obtained from lapachol, extracted and purified from the heartwood of Tabebuia sp. (Tecoma) and
further submitted to Antibacterial susceptibility and Minimal Inhibitory Concentration tests against clinical E. faecalis
strains obtained from Antnio Pedro University Hospital from Fluminense Federal University, which were originally
isolated from patients and donated to our laboratiories under research purposes. All molecular computations were
performed using SPARTAN 08 software. The structures were optimized to a local minimum and the equilibrium
geometry obtained in vacuum using AM1 semi-empirical methods. Subsequently, molecules were submitted to a singlepoint energy ab initio calculation, at the 6-31G* level, to calculate some stereoelectronic properties and perform the SAR
studies. Promising MIC values were identified for two triazole substituted compounds (1e = 8 lg/ml and 1c = 16 lg/ml) and
also for the non-substituted derivative (1a = 8 lg/m). In addition, molecular modeling studies pointed the low HOMO
energy values, HOMO density concentrated on the nor-b-lapachone ring, lipophilicity, solubility, and number HBA as
important stereoelectronic features for the antibacterial profile. The triazole compounds presented low theoretical toxicity
profile and drug-score higher than commercial antibiotics also fulfilling the Lipinski Rule of Five.
Conclusions:
Overall this work pointed the presence of the triazole groups as important but not essential for the activity of nor-lapachone derivatives. Our SAR analysis suggests the low HOMO energy values, HOMO density concentrated on the norb-lapachone ring, lipophilicity, solubility, and number HBA as important stereoelectronic features for the antibacterial
profile. In addition triazole compounds presented low theoretical toxicity profile and drug-score higher than commercial
antibiotics also fulfilling the Lipinski Rule of Five, which pointed them as promising candidates for further
studies in infections caused by multiresistant E. faecalis hospital strains.
Keywords: Enterococcus faecalis, Nor--Lapachone Derivatives, Potential Antibacterial Compounds
Financial Support: FAPERJ, CNPq, CAPES, UFF-PROPPi
Resumo:15-033
SOCIOECONOMICS PROFILE STUDY AND ASSOCIATIONS WITH THE TUBERCULOSIS CLINICAL FORM,
MULTIDRUG RESISTANCE AND TERAPEUTIC APPROACHES USED IN PATIENTS OF A PUBLIC CENTER TO
RESPIRATORY DISEASES.
Nascimento, A. A. 1; Pinheiro, C. G. 1; Pereira, L. S. R. 1; Santos, C. B. S. 1; Carvalho, J. S. M. 2; Carvalho,

F. L. Q. 1
1
Departamento de Cincias da Vida, UNEB
2
Departamento de Sade, Unijorge
Objectives:
Tuberculosis has important levels of morbidity and mortality among the contagious diseases in world, and is the main cause of
deaths by curable infectious diseases in adults. The low socioeconomic level inherent to most of these patients in treatment
reinforces the status of serious public health problem. In this work we aimed to study the socioeconomic profile of the patients
submitted to treatment and the possible associations with the clinical form of the tuberculosis, multidrug-resistance and
therapeutic approaches adopted in a public center to respiratory diseases in Salvador-BA.
All the data had been obtained from the medical records of 78 patients with 18 years old minimum hospitalized in the Octvio
Mangabeira Reference Hospital (OMRH), the public reference center to the respiratory diseases in this city, in 2010 with
tuberculosis diagnostic. This study was approved by the Ethics Committee of the Bahia State University (UNEB), protocol n.
0603100102672, as well as by the OMRH Center for Research Pneumology. From these data, it was possible the observe that
most of these patients were men, with age between 18 and 29 years old, low scholarity degree, and the maximum of one
minimum salary fee. More than 50% of them had no history of incomplete previous treatment, as well as were alcohol and
tobacco users. We also didnt detect any illicit drug users nor HIV carriers. 64% of those who still are in treatment use the
scheme 1 (izoniazide + riphampicin + pyrazinamide).Pulmonary tuberculosis was the predominant clinical form in the patients
studied. 4 % of the total 78 patients developed multidrug-resistance.
Conclusions:
From all the data obtained, we conclude that tuberculosis onset is strongly related to the socioeconomic conditions of life. The
low rate of multidrug resistance is related probably to the low level of abandoning the treatment, and to the unique scheme used
during the treatment. These are critical to the success of treatment.
Keywords: Pharmacological treatment, Socioecomics conditions, Tuberculosis
Financial Support: Financial Agency for the Support of Research of the State of Bahia (FAPESB).
Resumo:15-034
EVALUATION OF THE EFFECTIVENESS OF COSMETIC MESOTHERAPY FOR BODY CONTOURING
Soares, A. K. A. ; Maia, C. O. ; Azevedo, D. F. M. D. ; Lima, H. L. ; Raulino, G. P. ; Deocleciano, O. B.

UNIVERSIDADE DE FORTALEZA, UNIFOR
Objectives:
Mesotherapy is an invasive medical procedure that involves the application of intradermal injections and/or subcutaneous
injections of low concentrations of medicines directly into the area to be treated (Plast Reconstr Surg. 115:1425, 2005). This
method is indicated in the treatment of rheumatic diseases, sports medicine, dermatology and esthetics (Indian J Dermatol
Venereol Leprol. 73:60, 2007). In this context, this study is intended to evaluate the effectiveness of mesotherapy when used in
beauty treatments for body contouring.

This is a retrospective study of quantitative and descriptive character, from analysis of medical records of customers of a beauty
clinic located in Fortaleza-Cear. Data were collected between november to march this year, with a sample of 20 participants
from both sex. The evaluation of the effectiveness of mesotherapy was determined by reports of improvement of the doctor and
the patients described on the medical records. These parameters were based on the appearance of the treated area like, reduction
of measures, cutaneous sagging and of the fatty tissue. Also was used as criteria of evaluation, the comparison of waist measure
on the umbilical region (C1), 5cm above (C2), 5 cm below the same (C3), performed at the beginning and the end of treatment
using ANOVA as statistical test. The study was approved by the Research Ethics Committee of the University of FortalezaUNIFOR and is protected by resolution 196/96 of the National Commission of Research Ethics.The sample used in this study was
20 patients, 90% (n=18) of female and 10% (n=2) male. The mean age was 32.79 years (SD+10) and all patients were in
treatment for reduction of located fat. The treatment was performed completely by 60% of patients and the amount of
applications not mades per patient who left the treatment was 3.7 per session. The abandonment of 40% of cases were associated
with the discomfort of applications, mainly hematoma, pruritus and in some cases the availability of time of patients for the
weekly returns. The average loss measured after the treatment was -6.7cm in C1, C2 -5cm, and -2.5 in C3. Although not present
statistically significant differences the variations were considered satisfactory in most of the cases. One patient reduced 21 cm in
measure of C3. During treatment, there were reports of improvement, reported by the doctor in 90% and 70% for patients. About
19.04% (n= 4) reported improvement in sagging, 47.61% (n=10) in the measures, 9.52% (n=2) others and 19.04% (n=4) didnt
have information in the medical records. Mesotherapy in some patients was associated with other treatments such as physical
activity lymphatic drainage, endermologie, manthus, and ultrasound as mechanisms to enhance results.
Conclusions:
Positive results were found for mesotherapy to be on visual aspect of the treated areas such as in the required measurements of
treated areas. However, the success of treatment depends of individual variables, and their collaboration that goes from his
assiduity in applications to dieting, physical activity to enhance your results.
Keywords: INJECTIONS, INTRADERMAL, EFFICACY, MESOTHERAPY
Financial Support: PAVIC/UNIFOR
Resumo:15-035
ADVERSE EFFECTS CHARACTERIZATION OBSERVED IN TUBERCULOSIS PATIENTS.
Pinheiro, C. G. 1; Nascimento, A. A. 1; Pereira, L. S. R. 1; Santos, C. B. S. 1; Carvalho, J. S. M. 2; Carvalho,

F. L. Q. 1
1
Departamento de Cincias da Vida, UNEB
2
Departamento de Sade, Unijorge
Objectives:
The efficacy to the tuberculosis (TB) treatment is observed when there is at least six months of pharmacological therapy,
although the development of important adverse effects (AE) frequently observed. Such effects can alter many physiological
processes and had been related as important factors to patients give up the treatment. In this work we aimed to characterize the
adverse effects caused by the anti-TB treatment, and its relationship between the protocols used with the abandon to treatment
rate by patients with or without multidrug-resistance in a public center to respiratory diseases in Salvador-BA.
All the data had been obtained from the medical records of 78 patients with 18 years old minimum who were hospitalized in the
Otvio Mangabeira Reference Hospital (OMRH), the public reference center to the respiratory diseases in this city, in 2010 with
tuberculosis diagnostic, with or without multidrug resistance. This study was approved by the Ethics Committee of the Bahia
State University (UNEB), protocol n. 0603100102672, as well as by the OMRH Center for Research Pneumology. We observed
that most of the patients submitted to TB treatment developed any kind of AE (61,5%). Minor effects were observed in 37
patients (77,08%), in special attention to gastric manifestations (48,6%), headache and humor alterations (24,3%). 20 patients
(41,66%) had major effects, specially hepatotoxicity and other metabolic alterations (32,4%). We consider here that some patients
had either minor or major effects. When we analyzed all the protocols used (scheme I, scheme IR, personalized scheme and
multidrug resistant scheme) we observed that the number of patients showing any AE (48) was higher than those who didnt
relate anyone in all the situations (30). This high frequency of AE is related to the inherent toxicity of each drug used in the
treatment, as well as to the high impact caused when the patient abandon the therapy in any moment. When we studied the
relationship between the AEs with the treatment abandon, we observed that such events were more frequent in those who didnt
give up, since they were more exposed to the chemotherapics, and had higher probability to develop the AEs. Importantly, all the
patients who abandoned the treatment had developed any kind of AE. This means that the total number of patients suffering any
AE would be higher.
Conclusions:
Our data show that most of the TB patients studied developed minor adverse effects; also, in all the protocols used, more people
developed AEs than who did not, and that the onset of adverse effects was strongly related to therapy abandon. This work helps
us to the know about tuberculosis and the therapeutic management, better identify the patients that have critical profile, which
facilitate the development of politics of prevention to complications, improving the therapeutic and public control of the disease.
Keywords: Adverse effects, Pharmacological treatment, Tuberculosis
Financial Support: Financial Agency for the Support of Research of the State of Bahia (FAPESB).
Resumo:15-036
INTERACTION OF RISPERIDONE AND SERUM ALBUMIN. A SPECTROFLUORIMETRIC STUDY.
Fragoso, V. M. S. 1; Silva, D. 2; Cruz, F. A. O. 3; Cortez, C. M. 2

1
Faculdade de Cincias Mdicas, IBRAG - UERJ
2
Instituto da Matemtica e Estatstica, IME - UERJ
3
Departamento de Fsica, ICE - UFRRJ
Objectives:
Interaction mechanisms of risperidone to human (HSA) and bovine (BSA) serum albumins have been studied by using the
fluorescence quenching technique. Risperidone is an atypical antipsychotic drug used in many psychiatric disorders. The aim of
this study was to analyze the interaction of risperidone to HSA and BSA, characterizing the probable binding region, comparing
the results obtained for two albumins. The quenching curves were plotted, Stern-Volmer constants were estimated, and the nature
of the highest affinity binding sites of risperidone to the albumin were discussed.
Fluorescence measurements of risperidone with serum albumins were performed on a Hitachi - F3010 fluorescence equipment.
Quenching measurements were taken in 2ml of HSA and BSA 2x106M in 10 mM phosphate buffer, pH 7.4, when titrated with
risperidone (0.01 mM - 1.68 mM) at 25 C and 37 C. Using data from all experiments, graphs were plotted according to the
SternVolmer equation (Princ. of Fluores. Spectr. 3. ed. London: Plenum Press,2001). The graphs generated by the analysis of
the suppression of fluorescence were plotted from the arithmetic mean of three experiments for each temperature (standard
deviations lower than 10%), considering the average of two points around the maximum emission (max), max = 340 nm, for
BSA, and 338 nm, for HSA. In Stern-Volmer analysis, the cases for which the regression coefficient r2>0.9980 and p
Conclusions:
The decrease of Stern-Volmer constant caused by temperature elevation suggests the occurrence of static quenching for HSA
titrated by risperidone, interacting with this protein by forming complex risperidone - HSA. As the quenching intensity of BSA
was higher than of HSA, we are suggesting that the primary binding site for risperidone in albumin can be located close to
position 134 of the peptide chain in sub domain IB. At position 134 is a tryptophan residue in BSA.
Keywords: risperidone, albumin, antipsychotic drug, spectrofluorimetric
Financial Support: Fundao Carlos Chagas Filho de Amparo Pesquisa do Estado do Rio de Janeiro
Resumo:15-037
PRESYNAPTIC ADENOSINE AND MUSCARINIC RECEPTORS ARE INVOLVED IN HEXAMETHONIUM-, DTUBOCURARINE-, VECURONIUM-, AND ROCURONIUM-INDUCED TOFFADE.
1
1
1
2
1
Pereira, M. W. ; Bornia, E. C. S. ; Ambiel, C. R. ; Correia-de-s, P. ; Alves-do-prado, W.
1
Department of Pharmacology and Therapeutic., UEM
2
UMIB / Universidade do Porto, ICBAS
Objectives:
The quotient between muscular tension produced by the fourth stimulus (T4) and first stimulus (T1) is the train-of-four (TOF) ratio (Rtof=T4/T1) when the neuromuscular preparation is indirectly
stimulated at 2.0 Hz for 2 s. Reductions in Rtof values are referred to as TOFfade. TOFfade is determined by drug-induced blockade of facilitatory nicotinic receptors on motor nerve terminals. Here,
we investigated the participation of presynaptic inhibitory (M2 and A1) and facilitatory (M1 and A2A) receptors in TOFfade induced by hexamethonium (HEX), D-tubocurarine (D-TC), vecuronium
(VEC), and rocuronium (ROC).
The Ethics Committee for Experimental Studies of the State University of Maring approved the procedures (043-2007). The neuromuscular preparations of male wistar rats (250g) were assembled
according Blbring (Br J Pharmacol. 1:38, 1946). The preparations were indirectly stimulated at 0.2 Hz for 15 min, and TOF stimulation was applied at 15s intervals for 15min. Muscarinic antagonists
(pirenzepine, methoctramine) or adenosine receptor antagonists (DPCPX, ZM241385) were administered 1 min before the second sequence of TOF stimulation. The antinicotinic agents were
administered 1 min before the third sequence of stimulation. The diaphragm muscle was coupled to force displacement transducer (Grass FT 03, connected to Chart Software Powerlab Instruments).
The lowest concentration of HEX, D-TC, VEC or ROC able to produce 25% TOFfade 3 min after its addition in the bath were determined and administered after 10nM Pirenzepine (PZP), 1M
Methoctramine (MTC), 2.5 nM DPCPX or 10nM ZM241385 (ZM). Data were compared with ANOVA followed by the Bonferroni test (P1 antagonist) attenuated TOFfade produced by HEX
(5.00.4% n=6), D-TC (4.01.4% n=6), VEC (13.04.3% n=6), and ROC (7.01.8% n=6). MTC (M2 antagonist) fully prevented TOFfade caused by HEX (7.02.1% n=6). Surprisingly TOFfade
induced by D-TC (16.01.9% n=6), VEC (16.02.4% n=6) or ROC (152.4% n=6), was only partially attenuated by methoctramine. Antagonism of adenosine A1 receptors with DPCPX reduced
TOFfade caused by HEX (140.8% n=6), D-TC (18.02.8% n=6), VEC (11.01.7% n=6), and ROC (8.0 1.8% n=6). Blockade of the A2A receptor with ZM partially reversed TOFfade induced by
D-TC (12.03.3% n=6), VEC (11.01.7% n=6), and ROC (5.00.2% n=6), but not that caused by a pure neuronal nicotinic receptor antagonist, HEX, unless one increases the concentration to 50
nM (6.03.0% n=6).
Conclusions:
The results presented in this study add rationale for proposing an adequate use of subtype specific antagonists of muscarinic and adenosine receptors (like atropine- and metylxanthine-derivatives) in
patients safe recovery from neuromuscular transmission block caused by antinicotinic agents. We provided new information regarding the need for a distinctive pharmacological management of
patients receiving antinicotinic drugs acting preferentially at pre- versus post-synaptic sites, or both.
Keywords: TOF fade, Hexametonium, D-tubocurarine, Vecuronium, Rocuronium
Financial Support: Araucaria Foundation and FADEC-UEM.
Resumo:15-038
CHRONIC CAPTOPRIL TREATMENT PREVENTS ERECTILE DYSFUNCTION IN STREPTOZOTOCIN
INDUCED DIABETIC RATS
Neves, N. C. V. 1; Damasceno, E. C. 1; Batista, K. F. 1; Assis, L. V. M. 1; Guimares, H. N. 2; Grabeguimares, A. 1; Leite, R. 1

2
Escola de Engenharia, UFMG
1
Cipharma / Escola de Farmcia, UFOP
Objectives:
Diabetes mellitus when non-treated is commonly associated with many cardiovascular and neurological complications that could
lead to mild to severe male erectile dysfunction. The purpose of this study was to evaluate the beneficial effects of increasing
levels of circulating angiotensin-(1-7) [Ang-(1-7)] observed in chronic treatment with angiotensin-converting enzyme (ACE)
inhibitors such as captopril, on the erectile dysfunction (ED) that has been described in streptozotocin (STZ)-induced diabetic
male rats (Ethics Committee Protocol n. 2010/53).
Diabetic stage was induced by 3 consecutive injections of 40 mg/Kg of STZ in male Wistar rats (190-220g). Where considered
diabetic the rats presenting glucose level superior to 350 mg/dL 5 days after the last injection. Penile erection was evaluated in
control (normoglycemic), diabetic, and diabetic rats treated with captopril (25 mg/Kg) for 60 days after the injection of vehicle or
STZ. For that, the animals were anesthetized by ketamine/xylazine (100/14 mg/100g), had the major pelvic ganglion isolated and
electrically stimulated. Intracavernosal pressure (ICP) and mean arterial pressure (MAP) were measured and presented as an
index of erection (ICP/MAP). Frequency-response curves (1-12 Hz, 4V, 5ms pulse and 30 seconds for each frequency) were
obtained from rats of all groups. Erectile function was severely reduced in hyperglycemic compared to control normoglycemic
rats. Captopril treatment did not affect the ganglionic induced erectile response in normoglycemic rats, but significantly
improved, almost to a normal level, the impaired responses observed in diabetic hyperglycemic rats.
Conclusions:
Our data indicate that chronic treatment with ACE inhibitors such as captopril, that has been known to increase the circulating
levels of Ang-(1-7) and improve erectile function in spontaneously hypertensive rats, can also reverse the erectile dysfunction
observed in STZ-induced diabetic rats. We speculate that the high levels of Ang-(1-7) could be in part responsible for the
improvement of the erectile response in diabetic rats. These results can be very useful if applied to diabetic man who presented
ED.
Keywords: Angiotensin-(1-7), captopril, diabetes, erectile dysfunction
Financial Support: FAPEMIG, CNPq, UFOP.
Resumo:15-039
COMPARATIVE BIOAVAILABILITY OF BETAHISTINE TABLET FORMULATIONS ADMINISTERED IN
HEALTHY SUBJECTS
Arruda, A. M. M. ; Junior, P. P. D. ; Chen, L. S. ; Nucci, G. D.

Departamento de Farmacologia - Faculdade de Cincias Mdicas, UNICAMP
Objectives:
To asses the comparative bioavailability of two formulations (16 mg tablet) of betahistine (CAS 5579-84-0) in healthy volunteers
of both sexes.
The study was conducted using an open, randomized, two-period crossover design with a 1-week washout interval. Plasma
samples were obtained for up to 36 h psot dose. Plasma 2-pyridylacetic acid concentrations were analyzed by LC-MS/MS with
positive ion electrospray ionization using multiple reaction monitoring (MRM). From the 2-pyridylacetic acid plasma
concentration vs. time curves, the pharmacokinetic parameters were obtained for AUClast and Cmax.
Conclusions:
The limit of quantification was 4 ng/ml for plasma 2-pyridylacetic acid analysis. The geometric mean and 90% confidence
interval (CI) of test/reference percent ratios were: 98.94% (92.21% - 106.16%) for Cmax and 95.42% (91.74% - 99.25%) for
AUClast. Since the 90% CI for Cmax and AUCs ratios were all within the 80-125% interval proposed by the US Food and Drug
Administration Agency, it was concluded that the test formulation is bioequivalent to the reference for both the rate and the
extend of absorption.
Keywords: bioavailability, betahistine, pharmacokinetics, bioequivalence, LC-MS/MS
Financial Support: FAPESP, CnPQ, Apsen Farmaceutica S.A.
Resumo:15-040
QT INTERVAL AND ITS DRUG-INDUCED PROLONGATION
Souza, A. C. M. 1; Guimares, H. N. 3; Leite, R. 1; Sales, M. L. 2; Grabe-guimares, A. 1

3
DEE, Escola de Engenharia, UFMG
1
DEFAR, Escola de Farmcia, UFOP
2
DECME, Escola de Farmcia, UFOP
Objectives:
Describe the QT interval and its prolongation induced by drugs.
Literature research on Medline database using the keywords QT interval prolongation and cardiotoxic drugs. Theoretical
literature information was obtained after selection of articles that describe ECG alterations induced by drugs with different
therapeutic goals. According to FDA Guidance for Industry S7B Nonclinical Evaluation of the Potential for Delayed Ventricular
Repolarization (QT Interval Prolongation) by Human Pharmaceuticals when QT interval is prolonged, there is an increased risk
of ventricular tachyarrhythmia, particularly when combined with other risk factors (e.g., hypokalemia, structural heart disease,
bradycardia). Thus, much emphasis has been placed on the potential proarrhythmic effects of pharmaceuticals that are associated
with QT interval prolongation. The rapidly and slowly activating components of the delayed rectifier potassium current, IKr and
IKs, have the most influential role in determining the duration of the action potential, and so, the QT interval. The human ether-ago-go-related gene (hERG) and KvLQT1 gene encode pore-forming proteins KCNH2 and KCNQ1 that are thought to represent
the -subunits of the human potassium channels responsible for IKr and IKs, respectively. ECG abnormalities similar to those
seen in patients carrying mutations in the hERG gene can also be induced by direct blockade of hERG/IKr channels by a large
group of structurally diverse therapeutic compounds including many antiarrhythmics, antihistamines, antipsychotics and
antibiotics. The most common mechanism of drug induced QT interval prolongation is inhibition of the delayed rectifier
potassium channel that is responsible for IKr. At least three therapeutic compounds reduce hERG/IKr currents not by direct
blockade but by inhibition of hERG/IKr trafficking to the cell surface: (i) arsenic trioxide and and antimonial compounds, which
are used to treat acute promyelocytic leukaemia, (ii) the antiprotozoical agent pentamidine, and (iii) cardiac glycosides, which are
still used in heart failure. The antidepressive fluoxetine, represents the first member of a potentially large group of compounds
that directly block hERG and inhibit its traffic at the same time.(Biochemical Society Transactions 35:5, 2007).
Conclusions:
Therefore, it's important to investigate and understand the safety of therapeutic drugs that may induce arrhythmias due the QT
prolongation.
Keywords: cardiotoxic, drugs, prolongation, QT interval
Financial Support: FAPEMIG; UFOP; UFMG
Resumo:15-041
THERAPEUTIC EFFICACY STUDY OF MELAGRIO SYRUP IN PATIENTS WITH CLINICAL DIAGNOSIS OF
ACUTE BRONCHITIS.
Nascimento, D. F. ; Leite, I. O. ; Andrade, W. S. ; Leite, A. L. A. S. ; Fechine, F. V. ; Oliveira, J. C. ; Frota

Bezerra, F. A. ; Moraes, R. A. ; Moraes, M. E. A.
Depto. de Fisiologia e Farmacologia, UFC
Objectives:
Aim: Melagrio is a phytotherapic medicine composed of six medicinal plants with known action in the respiratory tract:
Mikania glomerata, Cephaelis ipecacuanha, Aconitum napellus, Polygala senega, Myroxylon balsamum and Nasturtium
officinale. The aim of this study was to evaluate the therapeutic efficacy of Melagrio in patients with clinical diagnosis of
acute bronchitis.

Methods and Results: A double-blind, randomized, parallel study, with 86 patients of both genders. The patients were randomly
divided in two groups: Melagrio (n = 43) or bromhexine (n = 43). They were all treated for 7 consecutive days using the
recommended dosage according to the treatment group and age. Clinical evaluations were performed on the pre-treatment and on
the seventh day of administration (post-treatment). To quantify the clinical response, we used a instrument composed of 12 items
related to signs and symptoms of respiratory infections (cough frequency, cough intensity, asthenia, nasal congestion, sore throat,
mucus elimination, anorexia, quality of sleep, headache , myalgia, fever and general condition), which were evaluated by using a
visual analog scale. In addition, we defined the general clinical evaluation as the arithmetic average of the 12 components of the
used instrument. In the analysis within the same group in all evaluated items, there was a significant improvement (P
Conclusions:
The Melagrio syrup showed that the improvement of the bronchitis symptoms was equivalent to the group treated with
bromhexine, thus proving the efficacy of this phytotherapic therapy.
Keywords: Clinical Trial, Therapeutic Efficacy, Bronchitis, Phytomedicine
Financial Support: CNPq, CAPES, FUNCAP, FINEP, MS-RNPC-UNIFAC-HM, and Instituto Claude
Bernard.
Resumo:15-042
BIOLOGICAL POTENCY EVALUATION AND CHARACTERIZATION OF RHEPO IN PHARMACEUTICAL
FORMULATIONS
Schutkoski, R. ; Camponogara, R. L. ; Freitas, G. W. ; Scheeren, L. E. ; Vecchia, F. D. ; Dalmora, S. L.

Departamento de Farmcia Industrial, UFSM
Objectives:
Erythropoietin (EPO) is the primary regulator of the rate of erythropoiesis. The recombinant human erythropoietin (rhEPO)
consists of a 165 amino acids polypeptide chain heavily glycosilated at three N-linked and one O-linked sites yielding a
molecular mass of 30-34 kDa. About 40% of the fully glycosilated EPO molecule consists of carbohydrate. Therapeutically,
rhEPO is used to stimulate red cell production by promoting maturation and growth of erythroid precursor cells. The aim of this
study was perform the characterization and the potency evaluation of erythropoietin in pharmaceutical formulations, by the in
vivo biological assay and physico-chemical methods.
Samples of pharmaceutical products from different manufacturers were used and the purity and molecular weight were assessed
using SDS/polyacrylamide gel electrophoresis and the heterogeneity by isoelectric focusing gels and immunoblotting of IEF gels
which showed a broad band in the 30.4 kDa range. The normocythaemic mice bioassay was performed using eight-week old
female BALB/c mice, which received a multiple daily injections of standard and sample solutions (3+3), during four days. The
blood sampling was performed 24 hours after the last injection and the reticulocytes counted by automated flow cytometry.
Parameters such as age, sex, strain, different injection schedules, and animals per group were evaluated using the multiple daily
injections protocol and the single injection protocol. The results were calculated against the Ph. Eur. BRP for rhEPO, giving
potencies within 91.70 and 115.40%. The pharmaceutical samples were also analyzed by the reversed-phase chromatographic
method which showed mean difference between the estimated potencies of 5.20% lower compared to the bioassay, with
significant correlation (P>0.05).

Conclusions:
The normocythaemic mice bioassay was validated and applied for the potency evaluation of rhEPO pharmaceutical formulations,
trying to develop alternatives in the context of the 3Rs. The results were also correlated to the chromatographic method
evaluating correlations which could contribute to improve the quality control and to assure their therapeutic efficacy of the
biological medicine.
Keywords: Characterization and potency evaluation, Erythropoietin, In vivo biological assay, Isoelectric focusing,
SDS/polyacrylamide gel electrophoresis
Financial Support: CNPq.
Resumo:15-043
PREVALENCE OF SELF MEDICATION IN ADULT POPULATION FROM THE URBAN AREA OF THE CITY OF
FLORIANO PI
Duarte, A. B. 1; Silva, D. J. S. D. 1,1; Santos, D. B. 1,1,1; Filho, M. D. S. 2; Martins, M. D. C. D. C. E. 2

1
Faculdade de Ensino Superior de Floriano, FAESF
2
Universidade Federal do Piau, UFPI
Objectives:
To assess the prevalence of self medication in the adult population of the urban area of the city of Floriano - PI the categories of
medicines most used and the motivations to use.
The survey consisted of cross-sectional study of population with sample of 552 people aged between 20 and 59 years. The adults
were selected through random sampling proportional to population in the neighborhoods. Data were collected through
questionnaires with closed and open questions. The significance level was set at 5% and statistical evaluation of variables was
accomplished using Chi-square test. The prevalence of self medication was 96.9%. The main reason for self-medication was pain
(78.6%). Drug group most widely used was represented by the analgesics drugs (48.5%). Previous experience with the
medication was the justification for self-medication pointed to by 64.6% of the participants. The protocols used were approved by
the ethics committee under number 2010/19.
Conclusions:
This study showed high prevalence of self-medication in the adult population of the urban area of Floriano-PI. The pain was the
main reason for self-medication, which explains the consumption of analgesics by a majority of respondents. Previous experience
with the medication was the main justification for self-medication. This demonstrates the need for preventive strategies in health
that clarify this population about risks f self-medication and contributes to the rational use of medicines.
Keywords: self medication, adults, prevalence
Resumo:16-041
EFFECT OF ANGIOTENSIN II ON SMOOTH MUSCLE FUNCTIONS IS MODULATED THROUGH
ALPHA1BETA1 INTEGRIN SIGNALING ACTIVATION
Moraes, J. A. 1; Dias, A. M. 1; Rodrigues, G. 1; Marcinkiewicz, C. 2; Assreuy, J. 3; Arruda, M. A. 4; Barjafidalgo, C. 1

1
Departamento de Farmacologia / Instituto de Biologia, UERJ
2
Neurovirology and Cancer Biology, Temple University
3
4
Diviso de Ensino, FIOCRUZ
Objectives:
The accumulation of vascular smooth muscle cell (SMC) in the atheromatous plaque is a key event of atherosclerosis. Under the
effect of different stimuli SMC migrate and proliferate, leading to formation of fibrous cap (atherosclerosis) or neointima
(restenosis). Angiotensin II (AngII), acting via its G-protein coupled receptor, AT1, is able to induce those effects through the
activation of PKC and ROS production, also involving the activation of integrin-mediated signaling pathways. In an attempt to
elucidate the molecular mechanisms involved in signaling pathways that modulate the effects of AngII on SMC functions.
The migration of SMC (A7r5 cell linage) towards AngII (100nM) was analyzed in Boyden chambers after 4h incubation. The
chemotactic effect of AngII was abolished when the cells were pretreated with the 11 integrin selective ligand, the disintegrin
Obtustatin (Obt 100nM). Stimulation of SMC with AngII induces a PKC-dependent FAK phosphorylation, uncoupling of ILK
to phosphorylated FAK and the association of ILK to actin, promoting cytoskeleton rearrangement. The treatment of cells with
Obt or with PKC inhibitor abrogated AngII effect on FAK phosphorylation, maintained the association between FAKILK(analyzed by western blot), inhibiting ILK association to F-actin, as observed by fluorescence microscopy in AngII-treated
SMC. The data indicate that, in SMC, the activation of 11 integrin-signaling pathway is downstream to PKC activation,
triggered by the binding of Ang-II to AT1-GPCR. Interestingly, the ROS production induced by Ang-II is more transient in cells
treated with Obt (DCF probe analysis). Additionally, it was demonstrated that the pretreatment with Obt induced G1 phase arrest
(cell cycle analysis) and diminishment of SMC proliferation (thymidine incorporation assay) of SMC treated with AngII.
Moreover, AngII-induced AKT phosphorylation and p21 expression reducement, is inhibited in cells treated with 11 integrin
inhibitor.
Conclusions:
In this work we suggest that 11 integrin may be an important target molecule to the development of more effective therapeutic
interventions in restenosis/atherosclerosis.
Keywords: ILK, Obtustatin, Angiotensin II, alpha1beta1 integrin, Smooth Muscle Cell
Financial Support: FAPERJ, CAPES, CNPq
Resumo:16-042
EFFECTS OF TREATMENT WITH RESVERATROL ON THE CONTRACTIONS STIMULATED BY KCL ON
ISOLATED AORTAS FROM HYPERTENSIVE RATS.
Antonieto, C. R. K. ; Oliveira, J. C. S. D. ; Mariinho, T. S. ; Scalabrini, A. C. ; Restini, C. B. A.

Medicine School - Universidade de Ribeiro Preto, UNAERP
Objectives:
Hypertension is a multifactorial clinical condition characterized by high and sustained levels of blood pressure. The vascular
endothelium develops important roles in physiologic and pathophysiologic hypertension. In experimental models to study the
hypertension the endothelium-dependent vasodilatation is severely reduced. The membrane of aorta rings isolated from rats
submitted to renovascular hypertension is more depolarized as compared to normotensive rats. The depolarization leads to the
activation of voltage-operated calcium channels (VOCCa2+) increasing the contractions in aorta from hypertensive rats, which in
general, are more sensitive to changes in Ca2+concentrations than in normotensive rats. Resveratrol, a polyphenolic compound
present in red wines has the ability to stimulate the vasodilatation due to the acute release of nitric oxide (NO) from vascular
endothelial cells. Aim: This study was designed to evaluate the participation of endothelial NO on the contractions stimulated by
depolarization induced with KCl in isolated aortic rings isolated from 2K-1C rats treated with resveratrol
Renovascular hypertension was induced in male Wistar rats (180g) by the implantation of a silver a Clip in a Kidney artery (2K1C). Control rats were submitted to abdominal laparatomy (2K). Resveratrol treatment (20 mg/kg, gavage) began one day after
the surgery and was performed tree times a week, during six weeks. After that, rats were killed by cervical dislocation and had
thoracic aortic isolated. The aortic rings with (E+) or without endothelium (E-) were set at a resting tension of 1.5 g in an organ
chamber containing Krebs solution, at 37C, pH 7.4. After testing the presence or absence of the E, concentration-response
curves (4.7-120 mM) for KCl were done before and 20 min after incubation 0.1M L-NAME (NOSynthase inhibitor). All
procedures and protocols were in accordance with the Guidelines for Ethical Care of the Experimental Animals; approved by
Institutional Animals Care and Use Committee (007/2010). Results were analyzed through One-way ANOVA; Newman-Keuls
post-hoc. Results: Maximum contractions were not altered in none of the conditions/groups of the reactivity studies. On the other
hand, the potency, evaluated by the negative log of concentration producing half the maximal effect values (pD2) was altered.
Presence of L-NAME increased (p
Conclusions:
Treatment with resveratrol does not alter the contractile response induced by KCl in aorta isolated from hypertensive rats. The
attenuation of the contractile response induced by KCl in aorta isolated from 2K-1C rats is dependent of the presence of
endothelium and of the NO production, but not to resveratrol treatment.
Keywords: hypertension, resveratrol, nitric oxide, endothelium, Potassium depolarization
Financial Support: CNPq, UNAERP
Resumo:16-043
ROLE OF ANG(1-7) IN THE CEREBRAL CIRCULATION OF 2K-1C HYPERTENSIVE RATS
Olivon, V. C. 1; Santiago, L. B. 1; Bonaventura, D. 2; Ramalho, L. N. Z. 3; Cortes, S. F. 2; Lemos, V. S. 1
ICB - Dep.Fisiologia e Biofisica, UFMG

2
ICB - Dep. Farmacologia, UFMG
3
FMRP - Dep. Patologia, USP
Objectives:
The renin-angiotensin system is fundamentally important for the development of renal hypertension. Ang (1-7) by activating the
receptor Mas produces relaxation, mainly via NO-cGMP. Hypertension can lead to a reduction in endothelium-dependent
vasodilation, caused in part by a reduction in the bioavailability of NO. The role of Mas receptors in the cerebral vascular
function is not yet well elucidated. The aim of this study was to investigate the role of Ang(1-7) in the carotid artery from 2K-1C
rats.
METHODS: We studied vascular reactivity in carotid artery rings from sham and 2K-1C Wistar rats. Ang (1-7) concentrationresponse curves were constructed in endothelium-intact or endothelium-denuded arteries pre-contracted with phenylephrine (0.10.03 mol/L) from sham and 2K-1C Wistar rats in the following conditions: 1) in the presence of A-779 (Mas selective
antagonist, 1mol/L); 2) in presence of PD 123,319 (AT2 selective antagonist, 1mol/L)+losartan (AT1 selective antagonist,
1mol/L)+HOE-140 (B2 selective antagonist, 1mol/L) or 3) in presence of L-NAME (non-selective NOS inhibitor, 0.1mol/L).
Proteins expression and localization were evaluated by Western blot, immunohistochemistry and immunofluorescence. All
procedures are in accordance with the standards and policies of the Animal Care Committee of UFMG (164/10). Data analyses:
ONE-WAY ANOVA. RESULTS: In endothelium-intact vessels Ang (1-7) elicited a concentration-dependent relaxation, which
was more pronounced in 2K-1C (Emax=70.85.2%**) animals as compared with sham (Emax= 45.665.12%). L-NAME
reduced but not abolished, Ang (1-7) relaxation in sham (Emax= 12.00.9%) and in 2K-1C (Emax= 30.3 0.18 %*) was still
increased in 2K-1C; A-779 also decreased Ang (1-7)-induced relaxation in 2K-1C (Emax= 40.2 3.8%*) and sham (Emax=
30.23.8%) but in the presence of A-779 the relaxant effect of Ang-(1-7) was still increased in 2K-1C. The blockade of AT1,
AT2 and B2 receptors evoked similar results as blockade of Mas; relaxation in response to Ang (1-7) was reduced in the presence
of the antagonists but remained larger in 2K-1C animals (Emax= 38.92.3%*) than sham (Emax= 22.41.2%). Endothelium
removal decreased Ang-(1-7)-dependent relaxation in sham (Emax= 29.03.1%) and 2K-1C (Emax= 50.63.7%*). However,
vasodilation remained bigger in 2K-1C vessels. In endothelium-denuded vessels, A-779 leveled Ang-(1-7)-induced relaxation
(sham=18.21.2%; 2K-1C=15.30.8%). We found similar results after simultaneous blockade of AT1, AT2 and B2 receptors
(sham=16.41.2%; 2K-1C=19.81.2%); L-NAME reduced Ang-(1-7)-induced relaxation in both groups (sham=10.41.0%; 2K1C=20.30.15%*). Mas receptor and endothelial NOS expressions were increased in 2K-1C group when to compare to sham
group. *p
Conclusions:
Together, our results show that in carotid artery of hypertensive animals, the vasodilator response to Ang-(1-7) is more
pronounced when compared to sham animals and mediated via Mas receptor. This increased vasodilator response is part mediated
by the endothelium with an important participation of NO. Another part of the increased vasodilator response is endotheliumindependent. Hence, it is possible that a compensatory mechanism mediated by Ang-(1-7) may exist to control the arterial
hypertension.
Keywords: Ang (1-7), carotid artery, hypertensive rats, Mas receptor
Financial Support: FAPEMIG and CNPq
Resumo:16-044
ROLE OF TLR4 IN BLOOD PRESSURE AND CARDIAC HYPERTROPHY OF SPONTANEOUSLY
HYPERTENSIVE RATS (SHR)
Echem, C. ; Bomfim, G. F. ; Dossantos, R. A. ; Fortes, Z. B. ; Carvalho, M. H. C.

Farmacologia / USP (ICB1), ICB/USP
Objectives:
AIM: Arterial hypertension is considered as a main risk factor for the development of cardiovascular disease (CVDs). As a
consequence, hypertension, leads to cardiac hypertrophy and a low degree chronic inflammation, with an increase in circulating
pro-inflammatory cytokines. Toll-Like Receptors (TLR) family was identified as one of the major factors involved in innate
immune response and its activation also induces pro-inflammatory cytokines production. Recently, TLR4 has been shown in
experimental models of atherosclerosis, heart failure, ischemic injury and septic cardiomyopathy indicating at with receptor could
be an important link between CVDs, inflammation and immune system activation. TLR4 expression was demonstrated to be
increased in heart with had undergone in a myocardial infarction, besides that TLR4-knockout mouse develop less severe cardiac
hypertrophy when compared to the wild type. Based on the evidences mentioned above, the purpose of this study was to
investigate the role of TLR4 in hypertension.
METHODS: SHR 5 and 15 weeks old (n=5) tail blood pressure (TBP) was measured by pletismography and cardiac TLR4
mRNA expression was evaluated by real time-PCR. Both parameters were also determined in SHR 15 weeks old (n=5), treated
with amlodipine (15mg/kgBW/day) and losartan (10mg/kgBW/day) for 15 days by gavage. Collagen I and III mRNA expressions
were also determined by real time-PCR in heart of SHR (n=6) untreated and treated with anti-TLR4 antibody (1ug/day, ip.) for 15
days. The wet and dry weight (mg) measurement was performed in the heart of SHR 15 weeks old (n=5) untreated and treated
with anti-TLR4 antibody. RESULTS: TBP (mmHg) of SHR 5 weeks old was 1082, and of SHR 15 weeks old was 1783. TBP
of SHR 15 weeks old treated with losartan (1382) and amlodipine (1425) was found lower than the untreated SHR. Cardiac
TLR4 mRNA expression of SHR 15 weeks old (1.00.13) was found higher than the SHR 5 weeks old (0.670.1).
Antihypertensive treatment decreased TLR4 mRNA expression when compared to the untreated SHR (untreated SHR: 1.00.13;
losartan: 0.480.17 and amlodipine: 0.530.12). The anti-TLR4 treatment decreased the mRNA expression of collagen I
(1.0960.04 vs. 0.6130.005) and collagen III (1.020.08 vs. 0.600.08) of SHR heart. This treatment also decreased the wet
weight (0.360.01 vs. 0.340.01) and dry weight (0.0880.001 vs. 0.0810.003) of SHR heart.
Conclusions:
CONCLUSION: We might suggest that TLR4 mRNA expression in the heart can be modulated by SHR blood pressure and
TLR4 seems to be involved in SHR cardiac hypertrophy.
Keywords: CARDIAC HYPERTROPHY, HYPERTENSION, TLR4
Financial Support: FAPESP and CAPES
Resumo:16-045
PHARMACOLOGICAL INHIBITION OF DIPEPTIDYL PEPTIDASE IV INCREASES GLUT4 PLASMA
MEMBRANE AND PROTEIN EXPRESSION IN SKELETAL AND CARDIAC MUSCLES OF SPONTANEOUSLY
HYPERTENSIVE RATS (SHR).
Salles, T. A. 1; Giannocco, G. 2; Oliveira, K. C. 2; Pacheco, B. P. M. 1; Girardi, A. C. C. 1

1
Instituto do Corao, HC-FMUSP
Faculdade de Medicina do ABC, FMABC
Objectives:
SHRs display changes on GLUT4 function, trafficking, and/or expression in insulin-sensitive tissues, suggesting that altered
regulation of this glucose transporter may account at least in part for the abnormalities of glucose metabolism in hypertension.
Inhibitors of the enzyme dipeptidyl peptidase IV (DPPIV) represent a novel class of antihyperglicemic agents that improve
glycemic control in type 2 diabetic patients. Furthermore, therapy with DPPIV inhibitors is associated with blood pressure
reduction. The present study was designed to test the hypothesis that pharmacological inhibition of DPPIV with sitagliptin may
improve glucose homeostasis in hypertensive animals by increasing skeletal and cardiac muscle GLUT4 expression and/or
plasma membrane (PM) translocation.
Studies were conducted in male SHR at 5 (Y) and 20 weeks (A) of age that were randomly divided into two groups and treated
twice a day, by oral gavage, with either sitagliptin (40 mg/day) (IDPPIV) or vehicle (water) for the period of ten days. Agematched Wistar Kyoto (WKY) rats served as normotensive controls. After sitagliptin treatment, Y-SHR + IDPPIV displayed
lower systolic blood pressure (SBP) than Y-SHR (119 3 vs. 136 4 mmHg; P < 0.05). In the A-SHR, sitagliptin treatment had
no significant effect on arterial blood pressure. Skeletal (soleus, EDL and gastrocnemius) and ventricular muscles were excised
from these experimental groups for analyses of GLUT4 expression and subcellular localization by immunoblotting. Y-SHR
exhibited less GLUT4 in the PM of the heart (-11.0 0.3%), soleus (-25.0 1.0%), EDL (-12.0 0.24%), and gastrocnemius (18.0 0.9%), as compared to Y-WKY. The reduction in PM GLUT4 expression in the heart, soleus and gastrocnemius were
accompanied by a slight, but significant decrease of total GLUT4 protein expression. The total protein abundance of GLUT4 in
these three types of muscles cells was normalized by the ten-day-treatment with sitagliptin. Interestingly, DPPIV inhibition
produced a significant increase (+20.0 0.48%) on PM GLUT4 expression in the ventricular myocytes from Y-SHRs + IDPPV
vs. Y-WKY. Reduction of GLUT4 PM in the skeletal and cardiac muscle cells of A-SHR vs. A-WKY was even more
pronounced: heart (-22.0 1.0%), soleus (-35.0 3.4%), EDL (-15.0 0.6%), and gastrocnemius (-30.0 1.8%). In the adult
SHR, lower PM GLUT4 expression was associated with lower GLUT4 total abundance in all muscle cell types analyzed.
Sitagliptin increased GLUT4 PM and total protein expression in the heart, soleus, EDL and gastrocnemius of A-SHR + IDPPIV
to levels higher than those found in the normotensive adult animals.
Conclusions:
Taken together, our data demonstrate that DPPIV inhibition increases GLUT4 expression and PM translocation in both skeletal
and cardiac muscles of SHR. Changes on GLUT4 expression by sitagliptin do not seem to correlate with blood pressure reduction
in this rat lineage. Moreover, our study has shed light upon a new mechanism by which DPPIV inhibitors improve glycemic
control.
Keywords: Dipeptidyl peptidase IV, Glut4, Heart, Hypertension, Skeletal Muscle
Resumo:16-046
THE INCREMENT IN NITRIC OXIDE MODULATION IN RAT CORONARY ARTERIES POST-MYOCARDIAL
INFARCTION IS A POSSIBLE MECHANISM INVOLVED IN PREVENTING THE ONSET OF HEART FAILURE
Couto, G. K. 1; Britto, L. R. G. 1; Mill, J. G. 2; Rossoni, L. V. 1

1
Department of Physiology and Biophysics/ ICB, USP
Department of Physiological Sciences/ CCS, UFES
Objectives:
The coronary circulation supplies the myocardium with oxygen and nutrients to maintain the cardiac function and then provides
the homeostasis. The endothelium, mainly by nitric oxide (NO) release, is able to adjust the coronary flow and it function has
been modified in some cardiovascular diseases, such as myocardium infarction (MI) and heart failure (HF). It is well known that
MI is followed by adaptive processes triggered in myocardium to maintain cardiovascular homeostasis which can retard the onset
of HF. We hypothesized that the coronary artery (CA) can be differently adjusted after MI in the presence and absence of HF.
Thus, we assessed the endothelium function in CA from infarcted rats with and without HF, focused in the nitrergic modulation.
Male Wistar rats, 10 weeks old, were anaesthetized (ketamine 50mg/kg plus xylasine 10mg/kg; i.p.) and submitted to left CA
ligation to produce MI or sham operation (SHAM; n=42). After 4 weeks, hemodynamic parameters were assessed in
anaesthetized rats and after the infarcted rats were subdivided in rats without (INF; n=32) or with HF (n=5). The results are
expressed as meanSEM. Statistical analyses: ANOVA (pvs. SHAM; #vs. INF). Afterwards, we evaluated the infarction area by
% of left ventricle (LV) area and the hypertrophy index by ratio ventricular weight/ tibia length. The infarction size was similar
between groups. The LV end diastolic pressure was increased after MI, however this increment was biggest in HF as compared to
INF (SHAM: 8.20.2 vs. INF: 10.50.3* vs. HF: 26.71.6 mmHg*#). The contractile index (dP/dt+) was reduced after MI, but
HF rats showed a greater decrease as compared to INF (SHAM: 7122160 vs. INF: 5960131* vs. HF: 4570125 mmHg/s*#).
On the other hand, MI reduced the relaxation index (dP/dt-) only in HF. The MI-induced right ventricular hypertrophy, but it was
greater in HF (SHAM: 5.10.1 vs. INF: 7.60.4* vs. HF: 11.00.7 g/cm*#). Afterwards, the septal CA was mounted on a wire
myograph and the endothelium-dependent (acetylcholine-ACh, 100M-100M) and -independent (sodium nitroprusside-SNP,
100M-10M) relaxation were evaluated. ACh-induced relaxation was decreased in CA of HF, while it was enhanced in CA of
INF as compared to SHAM (Emax: SHAM: 831 vs. INF: 911* vs. HF: 546 % relaxation*#). SNP-induced relaxation was
similar among groups. Further, some CA from SHAM and INF were incubated with NO synthase (NOS) inhibitor L-NAME (LN;
100M) to evaluate the nitrergic modulation on ACh relaxation. LN incubation reduced the ACh-induced relaxation in CA of
SHAM and INF rats, in addition it abolished the differences observed between groups in the absence of this inhibitor (Emax:
SHAM/LN: 378 vs. INF/LN: 366 % relaxation). Also, endothelial (e) and neuronal (n) NOS isoforms, Akt and AMPK protein
expression were evaluated by Western Blot in CA from INF and SHAM. The eNOS (79%), nNOS (52%) and AKt (45%) protein
expression were greater in CA from INF as compared to SHAM, whereas the AMPK protein expression did not change between
groups.
Conclusions:
After MI the CA showed different adjustments. The CA from HF rats presented endothelial dysfunction, while the endothelial
function in CA from INF rats was increased due to enhanced nitrergic modulation probably by activation of nNOS and AKteNOS pathways. This adjustment could act as a compensatory mechanism that improve the perfusion to non-infarcted
myocardium and could be involved on the prevention of HF installation.
Keywords: Coronary artery, Myocardial infarction, Heart failure, Endothelium, Nitric Oxide
Resumo:16-047
EFFECT OF PYRIDOSTIGMINE ON CARDIOVASCULAR AUTONOMIC CONTROL SIX TO SEVEN WEEKS
AFTER CORONARY ARTERY LIGATION
Sabino, J. P. J. ; da Silva, C. A. A. ; Fazan Jr. R. ; Salgado, H. C.

Depto. Fisiologia Universidade de So Paulo, FMRP-RP USP
Objectives:
Heart failure (HF) is characterized by remarkable changes in baro and chemoreflex control consisting of increased sympathetic
and reduced parasympathetic activity. While the sympathetic hyperactivity and its management in heart failure has been
thoroughly investigated, therapeutic options for reducing cardiac parasympathetic tone have not received the same attention.
Previous studies from our laboratory have demonstrated a protective effect of pyridostigmine (an acetylcholinesterase inhibitor)
on the autonomic control of arterial pressure and heart rate, due to the increase in parasympathetic function, 4 weeks after the
onset of heart failure in rats. However, the effect of pyridostigmine was not investigated during longer period of heart failure.
Thus, the present study evaluated the effect of pyridostigmine on the autonomic control of arterial pressure and heart rate, by
means of spectral analysis, six to seven weeks after coronary artery ligation.
HF was induced by coronary artery ligation of Wistar rats (250-300g) under anesthesia of ketamine/xilazyne (250 mg/kg, i.p.)
The procedure was similar for sham operated rats (Control), except that they did not undergo the ligation. After surgery the rats
received water or a solution of pyridostigmine (26 mg/kg/day, p.o.) during 6-7 weeks when the experiments were performed.
Beat-by-beat time series of systolic arterial pressure (SAP) were subjected to spectral analysis by Fast Fourier Transform. The
spectra of SAP were integrated into low (0.2 to 0.75 Hz) and high (0.75 to 0.75 Hz) frequency bands. The parasympathetic tonus
was evaluated by the tachycardic response caused by atropine (2 mg/kg, i.v.). HF rats exhibited reduced mean arterial pressure
(MAP) and no change in heart rate (HR) (96 2 mmHg, 341 9 bpm, n= 9) as compared to control rats (112 2 mmHg, 325 9
bpm, n= 12). Pyridostigmine impaired the decreased of MAP (105 4 mmHg, n= 4) of HF rats. In addition, HF rats showed
smaller LF power (1.3 0 mmHg2; n= 7) of arterial pressure as compared to control rats (3.1 0 mmHg2; n= 10).
Pyridostigmine prevented the decrease of LF arterial pressure power (4.4 1 mmHg2, n= 4) in HF rats. Parasympathetic tonus
was reduced in HF rats (30 6 bpm, n= 6) as compared to control rats (90 15 bpm, n= 7) while pyridostigmine protected the
parasympathetic tonus in HF rats (80 11 bpm, n= 3).
Conclusions:
These results demonstrate that pyridostigmine prevented the decrease in MAP, LF arterial pressure power and parasympathetic
tonus. Accordingly, pyridostigmine was efficient to protect the autonomic imbalance observed in HF rats.
Keywords: coronary artery ligation , parasympathetic tonus, pyridostigmine, spectral analysis
Financial Support: Capes, CNPQ and FAPESP
Resumo:16-048
EVALUATION OF NON-ENZYMATIC ANTIOXIDANT DEFENSE IN PATIENTS WITH METABOLIC SYNDROME
Martins, C. C. 1; Baldissarelli, J. 1; Ferreira, R. D. 1; Bagatini, M. D. 2,1; Cardoso, A. M. 1; Fiorin, F. S. 1;

Rodrigues, M. V. 1; Pereira, L. B. 1; Morsch, V. M. 1; Shetinger, M. R. C. 1
1
Universidade Federal de Santa Maria, UFSM
2
Universidade Federal da Fronteira Sul, UFFS
Objectives:
Metabolic Syndrome (MS) is a multifaceted condition, which is associated with a collection of disorders which contributes to
cardiovascular risk. This syndrome is often accompanied by oxidative stress, which occurs when reactive oxygen species
production is higher than the antioxidant capacity of the organism. Then, we evaluated the non-enzymatic antioxidant defense
that includes ascorbic acid (vitamin C) and Non-protein thiols (NPSH) in patients with MS to better characterize the presence of
reactive species in this pathology.
Thirty volunteer MetS patients were selected for this study and thirty control patients. They were aged between 35 and 65 years,
both sexes and all subjects gave written informed consent to participate in the study. The protocol was approved by the Human
Ethics Committee of the Health Science Center from the Federal University of Santa Maria, protocol number 98/10, Brazil. The
Non-protein thiols were assayed in plasma by the method of Ellman (Arch Biochem Biophys, v. 82, p. 70:77, 1959) and Vitamin
C analysis was made in serum by the method described by Jacques-Silva (J Biol Chem, v. 147, p. 399:407, 1943). Data were
analyzed statistically using test-t. Differences were considered significant when the probability was P < 0.05. We observed that
Non-protein thiols was significantly decreased in the MetS patients (0,60 0,15), when compared with the control group (1,15
0,33); the same was observed with Vitamin C (27,65 9,7 / 45,48 9,8).
Conclusions:
The vitamin C counters free radicals and regulates vitamin E metabolism by recycling oxidized tocopherols and its decrease
could be due to the increasing utilization of vitamin C as an antioxidant defense against reactive oxygen species. Similarly, nonprotein thiols, as GSH, play an important role in the antioxidant defense system by scavenging reactive species and regenerating
other antioxidants, as vitamin C. Its decrease may serve as a marker of increased oxidation and an indirect sign of higher
oxidative stress. Then, the elevated oxidative damage in MS patients can be evidenced by decreased the non-enzymatic
antioxidant defense.
Keywords: Metabolic Syndrome, antioxidant capacity , ascorbic acid , Non-protein thiols , cardiovascular risk
Financial Support: CNPq, CAPES, FAPERGS.
Resumo:16-049
ACUTE RENAL FAILURE EFFECTS IN THE GLOBAL PATTERN GENE EXPRESSION OF HEART TISSUE
Nakama, K. ; Abraho, M. ; Carneiro-ramos, M. S.

Objectives:
Patients suffering from renal failure are more likely to develop cardiovascular diseases, which aggravate patient health and can
lead to death. The heart is a target organ in many regulatory systems, such as hormones, neuronal and immunological systems,
modulating heart trophism directly or indirectly. This study aimed to determine changes in gene expression of hypertrophied
cardiac tissue in a renal ischemia-induced systemic inflammation model.
C57BL/6 young (20g to 26g), male mice were submitted to ischemic protocol, 60 min left renal pedicle unilateral occlusion
followed by reperfusion 5, 8, 15 or 20 days. Hearts were removed and total RNA was extracted and purified for the synthesis of
cDNA and cRNA. Analyses were performed using GeneChip Mouse Genome chip 430 2.0 Array (Affimetrix) containing 45,000
mice genes. The software dCHIP (DNA Chip Analyzer) was used to normalize and model expression. Gene modeled expression
cutoff was set to 20, comparable to modeled expression for TRbeta. The same software was used to separate genes that showed a
20% increase or decrease of the probes fluorescence intensity in comparison to the control group. The analysis results indicated
17,413 up-regulated genes and 18,297 down-regulated genes in at least one of the test groups with different days of reperfusion,
according to established criteria of 20% change of expression. Subsequently, we used the GenMAPP software to group the
selected genes in signaling pathways, being the analysis done with the same criterion of 20% genes up or down regulated.
Preliminary results indicate, in at least one of the groups tested, 405 up-regulated and 300 down-regulated pathways.
Conclusions:
The analysis of modulated genes indicate a higher number of down-regulated genes in test groups 5 and 8 days, as the largest
number of up-regulated genes were found in test groups 15 and 20 days. Concerning the regulated pathways, our data shows that
there is a progressive enhancement of the up-regulated pathways in test groups 5, 8 and 15 days, followed by a decrease in the
group 20 days. Thus, regarding the down-regulated pathways, the number of pathways negatively modulated in the group 8 days
is lower in comparison to the group 5 days. However, there was a subsequent increase in the groups 15 and 20 days.
Keywords: cardiac hyperthrophy, microarray, renal failure
Financial Support: FAPESP, CNPq e UFABC
Resumo:16-050
PRELIMINARY CARDIAC CHARACTERIZATION OF P2X7 PURINERGIC RECEPTOR KNOCKOUTS MICE
IMMUNIZED WITH PLASMIDS ENCODING THE CARDIAC MUSCARINIC RECEPTOR SUBTYPE M2
Martinez, C. G. ; Ribeiro, J. M. ; Bellio, M. ; Persechini, P. M. ; Kurtenbach, E.

Objectives:
Dilated cardiomyopathy (DCM) can be associated with the presence of auto-antibodies against the muscarinic receptor subtype
M2 (m2-AChR) as showed for Chagas disease patients and animals models immunized with M2-peptides or cDNA encoding for
M2-AChR. In these cases, the observed morphological and functional cardiac derangements was attributed to an increase in the
agonist action. Additionally an inflammatory response with an increase of fibrosis has been described, which in damage cells is
associated with the release of the pro-inflammatory mediator ATP, a ligand for purinergic P2X7 receptor in T cells. Thus, the
main purpose of this work is to determine if the absence of these receptors could interfere in the progression of DCM established
by the presence of anti-m2AChRs antibodies using C57Bl/6J knockout for P2X7 purinergic receptors.
For the development of DCM, C57Bl/6J mice, wild and P2X7 knockout, were immunized by biolistic with the plasmid
enconding for M2-AChR (pcDNA3-hM2). Other groups of animals, named controls, were immunized with the empty plasmid
pcDNA3. Twenty-three weeks after vaccination, cardio-respiratory function assessed by ergospirometry showed a significant
decrease in the time and speed of exercise in both groups of animals deleted for P2X7. All groups of animals showed an increase
in cardiac output that was dependent on an increase in heart rate. Analysis of regulatory T cells in the spleen through labeling for
Foxp3 showed no significant difference between those groups. However, the labeling for CD39, an ectonucleotidase
characteristic of immune cells, was increased in P2X7 knockouts vaccinated with plasmid pcDNA3 compared to the wild-type
mice group vaccinated with the same plasmid. Moreover the P2X7 knockout immunized with pcDNA3 group presents higher
levels of CD39 expression than P2X7 knockout immunized with pcDNA3-hM2. The last one present level comparable to wildtype mice vaccinated with pcDNA3 and pcDNA3-hM2. These dates could demonstrate a cross-talk beteween purinergic ang
muscarinic receptores. A few in vitro models demonstrated that might exist a compensatory action, which the absence of P2X7
can lead to an increase in function of CD39.
Conclusions:
We conclude that at this point, it was not possible to observe huge changes in the parameters analyzed, though the vaccination
promoted subtle immunological and functional changes, mainly in the cardiac output, frequency and expression of CD39.
Keywords: Cardiac muscarinic receptor subtype M2 , Dilated cardiomyopathy , P2X7 purinergic receptors
Financial Support: CAPES, CNPq and FAPERJ
Resumo:16-051
ECG ALTERATIONS INDUCED BY ANGIOTENSIN I AND ANGIOTENSIN II IN RATS
Silva, M. D. A. 1,1; de Paula, D. C. C. 1; Aguiar, L. D. 1; Grabe-guimares, A. 1; Serra, C. P. 1; Guimares,

H. N. 2,1
1
Universidade Federal de Ouro Preto, UFOP
2
Objectives:
Aim: Many experimental models were developed for the hypertension study and screening of potential therapeutic agents that
may contribute to the prevention and treatment of human hypertension. Induce hypertension via activation of RAS (Renin
Angiotensin System) is a classic model but the electrocardiogram (ECG) induced alterations are not described. The evaluation of
drug effects on QT interval of ECG is a fundamental component required for safety database, because its prolongations is
associated with arrythmias and sudden cardiac death. The aim of this study was to evaluate the pressor effect and its ECG
associated alterations at different doses of angiotensin I (ANG I) and angiotensin II (ANG II).
Methods: All the procedures were approved by the UFOP Ethical Committee (number 2010/83). Male Wistar rats (260 10 g)
were divided into 4 groups (n=6) that received: 1) Vehicle + ANG I; 2) Captopril + ANG I; 3) Vehicle + ANG II; 4) Losartan +
ANG II. The captopril or losartan (30 mg/kg) were orally administered 20 minutes before the animals were anesthetized with
quetamin/xilasin (100/14 mg/100g). Catheters were inserted into a femoral artery and vein, in order to obtain arterial pressure
(AP) signal and for drugs administration, respectively. Electrodes were inserted subcutaneously to obtain ECG signal (DII). After
surgical procedures, 3 doses of ANG I (1, 10 and 100 pmol) or ANG II (0,5; 5 and 50 pmol) were i.v. administered. The data
were analyzed with one-way ANOVA. Results: It was obtained significant and dose-dependent increase in blood pressure in
response to ANG I and ANG II. The increase of MAP was from 6 to 42 mmHg for ANG I, from 7 to 40 mmHg for ANG II. The
QT interval prolongation was significant for ANG I at 1 pmol (73,50 3,10 ms), 10 pmol (76,65 2,26 ms) and 100 pmol (78,73
2,80 ms) versus control (69,92 2,36 ms); and for ANG II at 5 pmol (74,54 2,96 ms) and 50 pmol (78,31 3,44 ms) versus
control (69,37 2,78 ms). The PR interval of ECG also presented significant increase after 1 pmol (57,17 1,04 ms), 10 pmol
(58,64 1,53 ms) and 100 pmol (60,73 1,05 ms) for ANG I versus control (54,96 1,12 ms); and after 5 pmol (58,67 3,00
ms) and 50 pmol (60,46 2,86 ms) for ANG II versus control (54,15 2,17 ms). The treatment with captopril reduced about 50%
and Losartan reduced about 80%. No changes were observed in the ECG, expected result for the acute treatment.
Conclusions:
Conclusion: The models of hypertension induced by ANG I and ANG II are also able to induce ECG alterations, mainly QT
interval prolongation. So they could be used for screening drugs with potential cardioprotective action.
Keywords: Angiotensin I, Angiotensin II, Cardioprotection, Eletrocardiogram, Hipertension
Financial Support: FAPEMIG, CNPq, UFOP
Resumo:17-135
TRPA1 RECEPTOR ACTIVITY EXPRESSED IN BLADDER DORSAL ROOT GANGLION NEURONS IS
REGULATED BY NEUTROPHIC FACTORS FOLLOWING SPINAL CORD INJURY
Andrade, E. L. 1; Ferreira, J. 2; de Castro, C. J. 3; Bento, A. F. 1; Souza, A. H. 3; Gomez, M. V. 3; Calixto, J.

B. 1
1
UNIVERSIDADE FEDERAL DE SANTA CATARINA, UFSC
2
UNIVERSIDADE FEDERAL DE SANTA MARIA, UFSM
3
UNIVERSIDADE FEDERAL DE MINAS GERAIS, UFMG
Objectives:
We previously observed that alterations in TRPA1 (Transient Receptor Potential Ankyrin 1) expression and activity in bladder
after spinal cord injury (SCI) contribute to the development of overactive bladder in rats (Am. J. Physiol. Renal Physiol., 2011).
Studies suggest that neurotrophic factors are involved in the regulation of TRPA1 sensitivity and expression (J. Clin. Invest., 115;
2393-2401, 2005; Dent. Res., 86; 550-555, 2007; Neurosci. Lett., 438; 221-227, 2008; Brain Res., 1284; 54-67, 2009). Therefore,
the aim of this study was to evaluate the influence of neurotrophic factors on the responsiveness and functionality of TRPA1 in
DRG neurons innervating bladder following SCI.
Spinal cords of anesthetized male Wistar rats (270-300g) were injured at T10 level by inserting a Fogarty 2F embolectomy
catheter (ethical committee process 000156). Dorsal root ganglia neurons (L6-S1 segment) of naive, sham-operated or SCI rats
were isolated 14 days after surgery and cultured in absence or presence of neurotrophic factors. Twenty-four hours later, the
calcium mobilization assay was performed. In other experiments, the neurotrophic factors levels in DRG neurons were evaluated
2, 7, 14 and 28 days after surgery. SCI induced increase in the proportion of DRG neurons responsive to TRPA1 agonist
cinnamaldehyde (300 M) (87%, 65/75 responsive cells/total cells) in comparison to non-operated group (naive) (68%, 30/44).
Moreover, SCI increased the calcium mobilization to cinnamaldehyde stimulus (72.7 21.9%; n = 5 experiments, 30-80 cells) in
comparison to sham-operated group. The exposure of DRG neurons of naive animals to neurotrophin BDNF (brain-derived
neurotrophic factor; 100 ng/mL) increased the proportion of DRG neurons responsive to cinnamaldehyde (89%, 80/90) in
comparison to control group (BDNF absence) (68%, 30/44). In addition, the exposure of DRG neurons of naive animals to NGF
(nerve growth factor; 100 ng/mL) or BDNF increased the calcium mobilization induced by cinammaldehyde (NGF: 93.3
25.3%; BDNF: 97.8 17.9%; n = 5 experiments, 30-80 cells) in comparison to control group. The NGF and BDNF levels were
significantly increased in DRG neurons on day 28 (2.5-fold) and on days 2 and 28 (3-fold) after SCI, respectively.
Conclusions:
Taken together, the results suggest that NGF and BDNF may be directly involved in the changes of TRPA1 responsiveness and
functionality induced by SCI. These events could contribute to the pathogenesis of overactive bladder following SCI.
Keywords: SPINAL CORD INJURY, BLADDER, NEUROTROPHIC FACTORS, OVERACTIVE BLADDER, RATS
Financial Support: CNPq, CAPES, FINEP, FAPESC, PRONEX
Resumo:17-136
BOTHROPS JARARACA HIGH MOLECULAR MASS KININOGEN (BJHK) INHIBITS CHANGES IN
LEUKOCYTE-ENDOTHELIAL INTERACTIONS INDUCED BY B. JARARACA SNAKE VENOM
METALLOPROTEINASES
Sanson, A. L. ; Zychar, B. C. ; Clissa, P. B. , ; Portaro, F. C. V. , ; Gonalves, L. R. C. ,

Instituto Butantan, IBU
Objectives:
Aim: The Bothrops jararaca (Bj) snake plasma is rich in protease inhibitors, some of which have inhibitory activity on toxins
from its own venom. One of these, which have a molecular mass of 110 kDa, is a potent inhibitor of cysteine proteases and
releases a peptide that induces contraction of homologous smooth musculature. For these characteristics this protein, named
BjHK (Bothrops jararaca High Molecular Weight Kininogen), was correlated to mammalian high molecular weight kininogens.
Moreover, it was found that this protein inhibits metalloproteases present in the Bj venom. Severe local inflammatory symptoms
are common in envenoming induced by B. jararaca snakebites, and It is known that metalloproteases is the main class of toxins
involved in the inflammatory activity of this venom. Our goal in this study was to test the effect of the BjHK on changes in the
leukocyte-endothelial interactions induced by Bj venom or by an isolated Bj metalloproteinase, in vivo, using intravital
microscopy.
Methods and Results: Bj venom (1 g) or Jararhagin (JAR) (0.5g), a hemorrhagic P3 metalloproteinase isolated from Bj venom,
incubated (30 min, 37 C) or not with BjHK (2 g) were injected (100L) into the scrotal bag of Swiss mice (25 to 28g body
weight, n= 5/ group). The microcirculation of cremaster muscle was analyzed 2 or 24h after the injections. We analyzed a
segment (100 m) of a post-capillary venule during 5 min and counted the number of adhered and emigrated leukocytes,
statistically comparing with the observed in control group injected with sterile buffered saline (significant when p< 0.05). The
results were the following: Adhesion 2h: control= 1.40.6; BjHk= 1.80.6; VBj= 12.20.6; JAR= 15.21.0; VBj+BjHK=
4.00.6; JAR+BjHK= 4.00.9; Adhesion 24h: control= 1.30.7; BjHK= 1.70.8; VBj= 4.20.4; JAR= 4.60.5; VBj+BjHK=
2.20.8; JAR+BjHK= 2.750.8; Emigration 2h: control= 1.00.3; BjHK= 0.60.4; VBj= 3.80.4; JAR= 6.40.5; VBj+BjHK=
2.60.6; JAR+BjHK= 1.70.5; Emigration 24h: control= 1.20.4; BjHK= 0.60.4; VBj= 14.40.6; JAR= 16.80.6; VBj+BjHK=
2.30.5; JAR+BjHK= 3.250.5. Results showed that BjHK did not induce changes in leukocyte-endothelial interactions, when
compared to the control group. In groups injected with Bj venom or with JAR, adhered and emigrated leukocytes were hugely
increased in all times studied. Adhered cells were significantly diminished after 24h of the venom or toxin injections when
compared to 2h, but emigrated cells were significantly increased. When incubated with BjHK, neither the Bj venom nor the JAR
induced significant changes in leukocyte-endothelial interactions in microcirculation when compared to the control group. This
effect was similar to the observed when Bj venom and JAR were treated with -phenanthroline, despite of the mechanism of
action of BjHK on the catalytic activity of JAR not be like a chelating agent.
Conclusions:
Conclusion: We conclude that BjHK can inhibit this inflammatory activity of the Bj venom probably by inhibiting the activity of
their metalloproteinases.
Keywords: kininogen, Bothrops jararaca, leukocyte, venom, metalloproteinases
Financial Support: FAPESP, CAPES, CNPq, and INCTTOX.
Resumo:17-137
CONSTITUTIVE NITRIC OXIDE SYNTHASES REGULATE NF-B NUCLEAR TRANSLOCATION AND SMOOTH
MUSCLE CELL ACTIVATION.
Scheschowitsch, K. ; Assreuy, J.
Dept of Pharmacology , UFSC
Objectives:
Cell stimulation with pro-inflammatory agents, such as lipopolyssacharide (LPS) and interferon- (IFN), causes cell activation
and tissue inflammation in vivo by increasing a variety of pro-inflammatory proteins through nuclear factor transcription kappa-B
(NF-kB) activation. During sepsis, a systemic inflammatory disease, a large amount of nitric oxide (NO) is produced in response
to cell activation and contributes to persistent hypotension. Results of our laboratory show that hypotension and mortality during
sepsis is prevented by early administration of NO synthase (NOS) inhibitors, suggesting that constitutive NOS play an important
role in sepsis. Vascular smooth muscle cells are the major component of vascular tonus maintenance and thus we decided to
investigate the importance of constitutive NOS in smooth muscle cell activation in vitro.
To evaluate cell activation a cell line of rat aorta smooth muscle cells (A7r5) were plated on a 96-well culture plate and
stimulated with LPS 1 g/mL and IFN 200 U/ml (LPS/IFN) for 24, 48 and 72 h. At each time period, 100 L of culture
supernatant was collected and nitrite concentration (M) determined by Griess reaction (n=3). Constitutive NOS activity was
investigated by plating A7r5 cells on a 96-well black culture plate, DAF-FM DA loading and stimulation with LPS/IFN in the
presence or the absence of NOS inhibitors (7-NI and L-NA, 200 M) or NO scavenger (PTIO, 100 M). Fluorescence intensity
was measured 30 minutes after stimulation and compared to control group and expressed as the percentage of fluorescence (n=3).
The effects of constitutive NOS inhibition on NF-kB nuclear translocation was evaluated by immunofluorescence. Cells were
plated in appropriate conditions and stimulated with LPS/IFN for 30 minutes, 2 and 4 hours. Images were obtained with confocal
microscopy (63x immersion) and quantified by LAS software. Results were expressed as the mean of fluorescence (in arbitrary
units, AU) in the nucleus (n=20 nucleus). All results are representative of 3 independent experiments SEM and statistical
comparisons performed by two-way ANOVA followed by Tuckey test. Nitrite concentration in stimulated cells was statistically
greater (3.7 0.3; 8.4 0.9; 13.9 0.4 M) compared to control group (0 M), respectively, in all the evaluated times. Cell
stimulation with LPS/IFN causes a NO pulse (3.7 0.5 versus control group 1.0 0.1), that was diminished in the presence of 7NI, L-NA and PTIO (2.5 0.3; 2.0 0.3; 1.4 0.3, respectively). Also, nuclear translocation of NF-kB in stimulated cells (56.3
0.7; 43.2 1.9; 14.9 0.5, respectively, to 30 minutes, 2 and 4 h) was diminished in the presence of these inhibitors compared
to stimulated cells in all the evaluated times (46.9 1.8; 37.1 1.5; 39.8 1.3, respectively, in the presence of 7-NI, L-NA and
PTIO, 30 minutes after stimulation). Reduced NO pulse and NF-kB translocation leads to diminished cell activation, as assessed
by nitrite concentration 48 h after cell stimulation.
Conclusions:
Smooth muscle cell stimulation with pro-inflammatory agents induces constitutive NOS activity and a NO pulse generation that
plays an important role in NF-kB nuclear translocation and cell activation.
Keywords: Nitric oxide synthases, NF-kB, smooth muscle cell
Financial Support: CAPES, CNPQ and FAPESC.
Resumo:17-138
EFFECTS OF RESVERATROL IN ANIMAL MODELS OF ACUTE NOCICEPTION INDUCED BY GLUTAMATE
AND CAPSAICIN IN MICE.
Bazzo, K. O. 1; Gomez, M. V. 3; Souza, A. H. D. 3; Campos, M. M. 1,2

2
Pontfica Universidade Catlica do Rio Grande do Sul, PUCRS
1
Mestrado em Medicina e Cincias da Sade, PPGMCS
3
Objectives:
Resveratrol (RSV) is a naturally-occurring polyphenol with well-characterized anti-inflammatory and antioxidant actions.
Moreover, significant antinociceptive effects have been demonstrated for RSV in the inflammatory hyperalgesia induced by
carrageenan in rats (Life Sci. 1317:21,2001). In the present study we have evaluated the effects of RSV in two acute models of
spontaneous nociception, induced by capsaicin and glutamate in mice.
Male Swiss mice (30 to 35 g, N=8 per group) were used. All the experimental protocols were approved by the local Animal
Ethics Committee (protocol number: 10/00175). The animals were pre-treated with RSV by oral route (50, 100 or 200 mg/kg), 30
min before the induction of nociception. In a separate series of experiments, RSV was administered intraplantarly (200 g/paw),
5 min beforehand. The spontaneous nociception was induced by the intraplantar injection of capsaicin (1.6 g/paw) or glutamate
(20 g/paw) into the right hindpaw. The time spent licking or biting the injected paw was registered during 5 and 15 minutes,
respectively. The results demonstrate that oral administration of RSV was able to significantly reduce capsaicin-induced
spontaneous nociception, at the doses of 50 and 100 mg/kg (2511% and 3610%, respectively). Furthermore, the oral treatment
with resveratrol (100 mg/kg) also produced a significant reduction of nociceptive behavior elicited by glutamate (399%).
Nevertheless, the local administration of RSV failed to significantly affect either capsaicin-induced nociception, whereas it
partially reduced the nociception induced by glutamate (33 9 %).
Conclusions:
The present findings suggest that RSV has antinociceptive effects in the acute nociception induced by both capsaicin and
glutamate, which appear to be related to an interference with central pathways of pain transmission. Additional experiments are
currently being performed to further characterize the mechanisms underlying the antinociceptive effects of RSV.
Keywords: Capsaicin, Glutamate, Nociception, Resveratrol
Financial Support: CAPES, CNpq and PUCRS
Resumo:17-139
LIPID BODY FORMATION INDUCED BY BOTHROPS JARARACA (BOTHROPOIDES), BOTHROPS INSULARIS
(BOTHROPOIDES), BOTHROPS ATROX AND BOTHROPS MOOJENI SNAKE VENOMS IN PERITONEAL
LEUKOCYTES OF MICE.
Nascimento, N. G. 1; Leiguez, E. J. 1,2; Giannotti, K. C. 1; Viana, M. D. N. 1; Teixeira, C. 1

1
Laboratrio de Farmacologia, Instituto Butantan - SP. , IBU
2
Universidade de So Paulo, USP
Objectives:
Lipid bodies (LBs) are cytosolic inclusions present in most eukaryotic cells, containing neutral lipids surrounded by a single
phospholipid membrane and specific proteins, such as the protein related to differentiation of adipocytes (ADRP). LBs are
involved in lipid metabolism and generation of inflammatory mediators. Moreover, increased numbers of LBs has been
demonstrated in inflammatory leukocytes. Venom from Bothrops genus snakes induce a marked inflammatory local reaction
characterized by leukocyte influx and release of a variety of inflammatory mediators. However, formation of LBs in leukocytes
present at the site of Bothrops venom injection is still unknown. In this study the ability of snake venoms from distinct species of
Bothrops genus, such as B. jararaca (BjV), B. insularis (BiV), B. moojeni (BmV) and B.atrox (BatxV) to induce LBs formation
and upregulation of ADRP protein expression was comparatively evaluated in mice peritoneal leukocytes.
Male Swiss mice were used (Butantan Institute Ethical Committee ref. 729/10). These animals received intraperitoneal (i.p)
injection of inflammatory doses of BjV (0.250 mg/g) or BmV (0.250 mg/g) or BiV (0.050 mg/g) or BatxV (0,025 mg/g) or
apyrogenic saline (control). At selected periods of time (1, 3, 6, 12, 24, 48 or 72 h) after each venom injection, the inflammatory
exudates were harvested to determine total number of leukocytes in Neubauer chamber and subtypes of leukocytes using Hema3
stained cell smears. LB formation was determined in macrophages stained with osmium tetroxide (1%) followed by counting
under phase contrast microscopy. ADRP protein expression was evaluated by Western blotting. Results showed that i.p injection
of BjV or BiV or BmV or BatxV caused a significant increase in the numbers of LBs from 1 up to 24 h (200%) when compared
with control cells (2.0 0.5 LB/cell, n=4), with no statistical difference among venoms from studied snake species. The highest
LB numbers were detected at the 12 h (300%) after venom injection in comparison with controls (2.4 0.7 LB/cell, n=4).
Moreover, all studied venoms caused a significant increase in levels of ADRP protein expression in leukocytes collected 12 h
after their injection.
Conclusions:
In conclusion, these data indicate the ability of Bothrops venoms either from the mainland or island to induce lipid body
formation and upregulate ADRP expression in leukocytes. Moreover, these data suggest that LBs have a role in the inflammatory
reaction induced by snake venoms from Bothrops genus.
Keywords: Snake Venom, Bothrops, Leukocytes, Lipid Body
Financial Support: FAPESP; CAPES.
Resumo:17-140
URB602N, AN INHIBITOR OF MONOACYLGLYCEROL LIPASE, DECREASES INFLAMMATION IN A MURINE
MODEL OF ACUTE LUNG INJURY
Costola-de-souza, C. ; Ribeiro, A. ; Ferraz-de-paula, V. ; Matazo, M. P. ; Palermo-neto, J.

Patologia Experimental e Comparada Universidade de So Paulo, VPT- FMVZ/USP
Objectives:
Recent studies indicate the importance of the endocannabinoid system, into different pathophysiological processes (Guindon,
Hohmann, 8:403,2009), where the endocannabinoids, such as 2-Arachidonoylglycerol (2-AG), modulate the immune response
(Cabral, Griffin-Thomas, 11:1, 2009). Based on these facts, the use of [1,1'-biphenyl]-3-yl-carbamic acid, cyclohexyl ester
(URB602) might be a nice opportunity to investigate the roles of 2-AG, since it blocks 2-AG degradation, making possible to
amplify 2-AG intrinsic actions. Thus, we analyzed the effects of URB602 under a neuroimmune perspective. Specifically, our
objective was to establish a dose-response curve for the anti-inflammatory effects of URB602 in a murine model of acute lung
injury.
Thirty-two C57BL/6 male mice were used; mice were divided randomly into six groups (n=4-6/group). Group 1 (G1) received
1.0mg/kg; group 2 (G2) 5.0mg/kg; group 3 (G3) 10.0mg/kg and group 4 (G4) 20.0mg/kg URB602 i.p. 60min prior a nasal
instillation of LPS (100ug/mL). Two Control groups received vehicle i.p. 60min prior to nasal instillation of LPS (GC1) or sterile
PBS (GC2). Each mice received 1ul of LPS/g bw. Twenty-four hours followed LPS or PBS nasal instillations, mice were
submitted to euthanasia and bronchoalveolar lavage fluid (BAL) as harvested for cell counting. Data were analyzed using Oneway ANOVA followed by Tukey-Kramer multiple comparison test. We observed that LPS treatment increases the number of
cells in the BAL compared to animals treated with PBS. Further statistical analysis showed that only animals treated with
20mg/kg of URB602 (F(5,19)=12,90;p,0,0001) showed no increases in the number of inflammatory cells induced by LPS in
BAL.
Conclusions:
We showed that URB602 (20mg/kg) decreases some inflammatory parameters in a murine model of acute lung injury.
Keywords: URB602, Acute lung injury, murine
Resumo:17-141
REACTIVITY OF LUNG FIBROBLASTS FROM SILICOTIC MICE: ANALYSIS IN 3D CELL CULTURE MODEL
Guimares-silva, A. M; Trentin, P. G. ; Dalzy, D. V; Perez, S. A. C; Martins, M. A. ; Silva, P. M. R

Objectives:
Silicosis is a chronic lung disease caused by the inhalation of crystalline silica particles, characterized by a slow and progressive
evolution leading to an intense fibrogenic response. Fibroblasts are considered crucial target cells in chronic fibrotic diseases and
over the past years, the development various types of 3D culture systems have garnered much attention and gained increased
recognition as important research tools. In this study we evaluated the reactivity of lung fibroblasts from normal or silicotic mice
using a 3D cell culture (spheroid) assay in vitro.
Male Swiss-Webster mice were instilled with intranasal crystalline silica particles (10 mg/50 L) or with the same volume of
saline. After 7 days, lungs were perfused with PBS and digested with enzyme (collagenase A) to obtain the fibroblasts. The cells
were cultured in DMEM + fetal bovine serum (10%), and after they entered in confluence they were replicated three times. Cells
were put in a 96 wellplate, previously covered with agarose, in a density of 1.25x104 cells/well. The analysis was performed in
the period of 1-4 days and included parameters such as size/diameter and cell proliferation, evaluated by image analyses (Image Pro Plus program) and thymidine [H3] uptake, respectively. All experimental procedures were performed in accordance with the
guidelines of the Committee on Use of Laboratory Animals of the Oswaldo Cruz Foundation (L-034/09).The growth kinetics
revealed that at early time-points (1-2 days), the spheroids obtained from normal mice fibroblasts presented lower density and
higher size/diameter as compared to those at later time points (3 - 4 days). Spheroids from cells obtained from silicotic mice
exhibited a similar profile, though showing themselves bigger than those from normal mice. The proliferation rate was also
increased in the case of fibroblasts from silicotic mice as compared to that of normal cells (192.315.6 vs. 11317.4,
respectively) (mean SEM; n=6). When the two spheroid populations were submitted to stimulation with the pro-fibrotic
cytokine IL-13 (40 ng/mL), for 1-4 days, we noted that cells were activated as attested by a significant increase of size/diameter
and of proliferation rate. Notably, under condition of IL-13 stimulation, silicotic spheroids presented higher size/diameter and
proliferation rate when compared to normal spheroids (rate proliferation: 1373147.3 vs. 630117.8; respectively).
Conclusions:
Altogether, our results show that fibroblasts from adult mice lungs can be cultured in a 3D culture system and that the spheroids
generated from silicotic fibroblasts were more activated than the normal ones. The 3D model for lung fibroblasts seems to be a
quite reproducible and easy-handling culture, representing a promising assay for the search of anti-fibrotic drugs.
Keywords: silicosis, lung fibroblasts, 3D culture
Financial Support: FIOCRUZ/CNPq/FAPERJ
Resumo:17-142
REGULATORY ROLE OF THE PROTEIN ANNEXIN-1 ON THE LIPOPOLYSACCHARIDE INDUCED LUNG
INJURY IN MICE
Matheus-souza, D1; Arantes, A. C. S1; Ferreira, T. P. T1; Trentin, P. G. 1; Pires A. L. A1; Flower, R. 2;
Perretti, M. 2; Martins, M. A. 1; Silva, P. M. R 1
2
Dept.of Biochemical Pharmacology,The William Harvey Institut, WHI
1
Fundao Oswaldo Cruz, FIOCRUZ
Objectives:
Introduction: Escherichia coli lipopolysaccharide (LPS) induces an acute lung injury characterized by vascular permeability
increase and leukocyte infiltration in the air space and lung parenchyma, leading to impairment of the respiratory function.
Glucocorticoids are considered as critical agents based on their anti-inflammatory properties, which is shown to be at least
partially dependent on the induction of an intermediate protein called annexin-1. In this study we aimed to investigated the
potential regulatory role of annexin-1 on the acute lung injury caused by LPS in mice.

Methods: Anesthetized male Balb/c (ANX1+/+) mice and annexin-1 knockout mice (ANX1-/-) received a single intranasal
instillation of LPS (25g/ 50 L) or saline. The analyses were made 24h-post LPS provocation. Lung function (resistance and
elastance) and airways hyperreactivity to methacholine (3 - 27 mg/mL) were evaluated by whole body invasive plestimography
(Finepoint, Buxco System). Total and differential leukocytes in bronchoalveolar lavage (BAL) were counted in Neubauer
chamber and cytospin preparations stained with May-Grunwald-Giemsa dye, respectively. Collagen was measured by Sircol
technique and cytokine/chemokine generation by ELISA. All experimental procedures were performed in accordance with the
guidelines of the Committee on Use of Laboratory Animals of the Oswaldo Cruz Foundation (L-034/09). Results: ANX1+/+
mice when stimulated with LPS showed increased levels of basal lung resistance and elastance as well as airways hyperreactivity
to aerosolized methacholine. A marked inflammatory response was noted in LPS-provoked mice, including leukocyte infiltration
(mainly neutrophils) in the BAL fluid as well as generation of chemokines (KC and MIP-2) and cytokine (TNF) in the lung
tissue. In the case of ANX1-/- mice, an exacerbation of lung function compromise, leukocyte infiltration in the BAL fluid and
cytokine/chemokine generation in the lung tissue was evidenced. A marked increase in the tissue collagen production was also
detected in the ANX1-/- animals as compared to ANX1+/+ mice (from 47,43 3,566 to 26,42 2,848 g/right lung,
respectively. Mean SEM, n=6)
Conclusions:
Conclusion: Altogether our data show that the annexin-1 seems to effectively regulate the inflammatory response caused by LPS
in mice, indicating that treatment with this protein or even with its derivatives could be an important pharmacological tool for the
treatment of inflammatory diseases such as acute lung injury.
Keywords: Annexin-1, Inflammation, Lipopolysaccharide, Lung, Mice
Financial Support: Financial Support : FIOCRUZ, CNPq, FAPERJ and CAPES.
Resumo:17-143
ANTI-INFLAMMATORY ACTIVITY OF COCOS NUCIFERA L. (ARECACEAE), GIANT VARIETY
Silva, R. R. 1; Silva, D. O. 1; Zardo, R. S. 2; Gonalves, M. R. 2; Alviano, C. S. 1; Fernandes, P. D. 2;

Alviano, D. S. 1
1
Instituto de Microbiologia Paulo de Ges (UFRJ), IMPPG
2
Instituto de Cincias Biomdicas (UFRJ), ICB
Objectives:
The fruit of Cocos nucifera L. (Arecaceae) has a fiber husk rich in polyphenolic compounds that is popularly used to several
disorders. Recent results show that aqueous extracts from husk fiber of C. nucifera present different biological activities. In this
study our objectives were to evaluate the anti-inflammatory activity of a variety of C. nucifera that has not been investigated yet.
The Cocos nucifera L. (Arecaceae), giant variety, was collected in Aracaju, Brazil (in May, 2007). The husk fiber (414 g) was
dried in the sun, finely ground and the powder soaked for 3 h in 6 L of boiling distilled water. Then, the extract was filtered,
lyophilized and stored at 200C, yielding around 10% of the dry weight of starting material. The aqueous extract was
resuspended in distilled water in all the experiments. The anti-inflammatory activity was evaluated in models of abdominal
writhing induced by acetic acid (2%, ip), formalin-induced licking response (2%, intraplantar) and subcutaneous air pouch (SAP)
induced by carrageenan (1%) injection. Swiss mice (20-25 g, n = 5-6) were orally pre-treated with the aqueous extracts from C.
nucifera (doses 50, 100, and 150 mg/kg). The use of animals was approved by the ethical committee of animal experimentation
from Centro de Cincias da Sade (UFRJ) and received the number ICBDFBC015. The results are presented as the mean S.D.
and statistical analysis was ANOVA followed by Bonferroni (*p
Conclusions:
Our results suggest that the aqueous extracts from husk fiber of Cocos nucifera L. (Arecaceae), giant variety, have significant
anti-inflammatory activity. Since the husk fiber is an industrial reject, the results obtained in the present study suggest that it
might be a very inexpensive source of anti-inflammatory drug that warrants further investigation.
Keywords: anti-inflammatory activity, Arecaceae, Cocos nucifera, extract, giant
Financial Support: CAPES, CNPq, and FAPERJ.
Resumo:17-144
AGLUTININ ISOLATED FROM THE INVERTEBRATE MARINE HOLOTHURIA GRISEA (HGA) REDUCES
INFLAMMATORY HIPERNOCICEPTION
Luz, P. B. 1; Arago, K. S. 1; Osrio, C. B. H. 2; Moura, R. D. M. 1; Melo, A. A. D. 1; Carneiro, R. F. 1;

Cavada, B. S. 1; Alencar, N. M. N. D. 1
1
Departamento de Fisiologia e Farmacologia/ Faculdade de Medi, UFCe
2
Departamento de Bioqumica e Biologia Molecular/ Centro de C, UFCe
Objectives:
The present work aimed to evaluate the effect of lectin's Holothuria grisea (HGA) in the model of mechanical
hipernociception induced by carrageenan (Cg).
Animal handling and experimental protocols were approved by the Institutional Ethics Committee under number 60/09.
Male Swiss mice (n=6, 25-30g) were placed in acrylic cages (122017cm) with wire grid floors, 1530 min before the
beginning of the test. The test consisted of evoking a hindpaw flexion reflex with a hand-held force transducer adapted
with a 0.5mm polypropylene tip. The investigator was trained to apply the tip perpendicularly to the central area of the
hindpaw with a gradual increase in pressure. The end point was characterized by the removal of the paw followed by
clear flinching movements. After the paw withdrawal the intensity of the pressure was automatically recorded, and the
value for the response was obtained by averaging four measurements. The animals were tested before (T=0) and after
treatment (1, 2, 3 and 4 hours). HGA (1 e 10mg/kg) or saline were administred i.v. 30 min before injection of Cg in right
hind paw (300g/paw). For measurement of myeloperoxidade activity (MPO) the mice were treated with HGA 10mg/kg
i.v. or saline, 30 min later they received 300g/paw of Cg, 4h later they were sacrificed and a sample of tissue (50mg) has
been removed. The results were presented as number of neutrophils106/mg tissue (MPO activity). Nitrite (NO2 )
concentrations were determined in serum obtained from the animals 4h after intraplantar injection of Cg. HGA (10
mg/kg) or saline was administered i.v. 30 min before Cg. The results were expressed as micromoles of nitrite. Regarding
statistics we used ANOVA/Bonferronis test. P
Conclusions:
The results indicate that HGA shows a potent antinociceptive effect on mechanical inflammatory hypernociception
evoked by Cg may be associated with the inhibition of neutrophil migration and NO production.
Keywords: AGLUTININ, HIPERNOCICEPTION, INFLAMATION, INVERTEBRATE
Resumo:17-145
ROLE OF TRPA1 RECEPTOR IN NOCICEPTION INDUCED BY MONOSODIUM URATE (MSU) CRYSTALS IN
RATS.
Trevisan, G. ; Hoffmeister, C. ; Rossato, M. F. ; Ferreira, J.

Departamento de Qumica/Universidade Federal de Santa Maria, UFSM
Objectives:
Gout is characterized by the deposition of monosodium urate (MSU) crystals in joints. Despite being one of the most painful
forms of arthritis, the mechanisms responsible for acute painful gout attacks are poorly understood. Recently, our research group
has shown the involvement of TRPV1 receptors in the acute nociception and edema evoked by MSU in rats. As TRPA1 receptor
are co-expressed with TRPV1 in capsaicin-sensitive fibers, the present study aimed to investigate if TRPA1 receptors also
contribute to the nociceptive and edematogenic response induced by MSU crystals in rats.
Adult male Wistar rats (200-300 g) were used. We observed the signs of spontaneous nociception, cold alodynia and edema in the
animals produced by MSU (0.25 mg/paw, 100 l) administered subcutaneously (s.c). Separate groups of animals received instead
the s.c. injection of the vehicle (PBS). Animals were placed individually in chambers and adapted for 20 min before injection.
The numbers of flinch responses were observed during 10 minutes after the injection and it was recorded and considered as
indicative of spontaneous nociception. Cold allodynia was observed as a score (0= non response, 1= flinching, 2= shaking,
4=licking of the paw when 50 uL of acetone was delivery). The edema formation was assessed as an increase in paw thickness
measured by a digital calipter, 15 min after s.c. injection and the results were expressed as the difference between the basal value
and the test value. HC0300313 (100 ug/paw) and camphor (150 nmol/paw), a selective and a non-selective TRPA1 antagonist,
respectively, were also subcutaneously injected with MSU. Allyl isothiocynate (AITC, 1 nmol/paw) a TRPA1 agonist, was used
as a positive control in all experiments. To further explore the role of capsaicin-sensitive fibers in the nociceptive and
edematogenic effect induced by MSU, animals were submitted to a perineural capsaicin desensitization, were the sciatic nerve
received a 10 L injection of 2% capsaicin or vehicle (10% ethanol, 10% Tween 80 and 80% PBS). After 7 days, animals were
submitted to a subcutaneous injection of MSU, AITC or PBS. Treatment with the selective TRPA1 receptor antagonist
HC030031 inhibited spontaneous nociceptive responses to MSU and AITC (Imax of 876 and 6173 %, respectively). Camphor
also reduced MSU-induced nociceptive response induced by MSU and AITC (Imax of 749 and 755 %, respectively). The cold
allodynia induced by MSU or AITC (946 and 928, respectively) was also effective reduced by the treatment with TRPA1
antagonists HC 030031 and Camphor (I max 928 and 928, respectively).The edema formation induced by MSU or AITC was
also reduced by camphor (Imax of 6517 and 833 %, respectively) or HC030031 (I max of 7912 and 6716 %, respectively).
The nociception and edema induced by AITC or MSU were largely inhibited in 765 and 884 or 924 and 6812%,
respectively, when animals were submitted to a perineural capsaicin desensitization protocol.
Conclusions:
Collectively, the present findings demonstrate that MSU produces nociceptive and edematogenic responses through, at least in
part, TRPA1 receptor activation.

Keywords: Gout, Nociception, TRPA1
Financial Support: CNPQ, CAPES
Resumo:17-146
CANNABIDIOL PRODUCES A LONG-TERM ANTI-INFLAMMATORY EFFECT IN A MURINE MODEL OF
ACUTE LUNG INJURY: ROLE FOR A2A ADENOSINE RECEPTOR
Ribeiro, A. 1; Ferraz-de-paula, V. 1; Pinheiro, M. L. 1; Almeida, V. I. 1; Quinteiro-filho, W. M. 1; Akamine,

A. T. 1; Hallak, J. E. C. 2; Zuardi, A. W. 2; Crippa, J. A. 2; Palermo-neto, J. 1
1
Departamento de Patologia / FMVZ-USP, FMVZ-USP
2
Departamento de Neurociencias e Comportamento FMRP-USP, FMRP-USP
Objectives:
Endocannabinoid system has become a topic of great interest in pharmacology due to its remarkable distribution in mammal
organisms and capacity to play a modulatory role on diverse physiological functions, including immune system modulation.
Acute lung injury (ALI) is a murine model of Acute Respiratory Distress Syndrome (ARDS), a condition developed by many
patients with sepsis, which affects thousands of people every year. We have previously shown that cannabidiol (CBD) decreases
leukocyte migration into the lungs and TNF concentration in the bronchoalveolar lavage fluid on ALI. Here our goal was to
analyze the effects of CBD in the progression of ALI and the mechanism of action involved.
C57BL/6 male mice were used and divided randomly in 3 groups: vehicle+PBS, vehicle+LPS, and CBD+LPS (n=5-7/group).
Experimental groups received CBD (20 mg/kg) i.p. 60 min prior to nasal instillation of LPS (100 ug/mL). Control groups
received vehicle i.p. 60 min prior nasal instillation of LPS or sterile PBS. Each mouse received 1 uL of LPS or PBS per g of
mouse weight. One, two, four, and seven days after LPS or PBS nasal instillation, mice were submitted to euthanasia and
bronchoalveolar lavage fluid (BAL) was harvested for cell counting and cytokines/chemokines measurement, and a lung fraction
was collected for myeloperoxidase (MPO) analysis. Moreover, an antagonist of A2A adenosine receptor (ZM241385) was used
(30 min prior to CBD treatment) to investigate the involvement of adenosine signaling in the effects observed. Data was analyzed
using One-way ANOVA followed by Tukey-Kramer multiple comparison test or Kruskal-Wallis followed by Dunns multiple
comparison test. We show that vehicle+LPS group presented a remarkable increasing on BAL cell counting following 1 (F (2,13)
= 23.58; p < 0.0001), 2 (F (2,16) = 36.83; p < 0.0001), and 4 (F (2,17) = 17.27; p < 0.0001) days of the induction of lung
inflammation. We observe that CBD treatment decreased BAL cell counting following 1, 2, and 4 days of the induction of
inflammation. We also show that CBD treatment decreased cytokine/chemokine concentration in the BAL, such as TNF (day 1)
(F (2,12) = 11.38; p < 0.001), IL-6 and MIP-2 (day 2) (F (2,14) = 6.33; p < 0.01 and KW = 10.56; p < 0.01, respectively).
Moreover, we show that CBD treatment decreased MPO activity following 1 (F (2,12) = 16.39; p < 0.001), 2 (F (2,18) = 17.21; p
< 0.0001), and 4 (F (2,17) = 49.55, p < 0.0001) days of the induction of lung inflammation. Notably, when mice received
ZM241385, CBD was not able to decrease BAL cell count as previously observed on day 1, 2, and 4 after the induction of
inflammation.
Conclusions:
We show that a single CBD treatment is able to produce a long-term decrease on several inflammatory parameters in a model of
ALI. Partially, we also show that A2A adenosine receptor is involved on those effects. Therefore, we believe that CBD might be
a useful therapeutic tool in the treatment of inflammatory lung diseases.

Keywords: Cannabidiol, Acute lung inflammation, Adenosine receptor
Financial Support: FAPESP (2009/51886-3) and CNPq.
Resumo:17-147
NEURAL MOBILIZATION REVERSES PAIN BEHAVIORAL AND INCREASE PROTEIN ZERO (PO)
EXPRESSION IN SCIATIC NERVE AFTER NEUROPATHIC PAIN IN RATS
Santos, F. M. ; Giardini, A. C. ; Silva, J. T. ; Chacur, M.

Departamento de Anatomia, Universidade de So Paulo, USP
Objectives:
The neural mobilization (MOB) technique is a noninvasive method that has proved clinically effective in reducing pain
sensitivity and consequently in improving quality of life after neuropathic pain. The present study examined the effects of neural
mobilization on pain sensitivity induced by chronic constriction injury (CCI) in rats. In addition, we also evaluated the protein
zero (PO) expression in sciatic nerve after neuropathic pain and MOB treatment.
The CCI was performed on adult male rats, submitted thereafter to 10 sessions of MOB, each other day, starting 14 days after the
CCI injury. Over the treatment period, animals were evaluated for nociception using behavioral tests, such as tests for allodynia
and thermal and mechanical hyperalgesia. At the end of the sessions, the sciatic nerve was analyzed using Western blot assays for
neural protein zero (PO). The results showed that MOB treatment induced an early reduction (in the second session) of the
hyperalgesia and allodynia in CCI-injured rats, which persisted until the end of the treatment. On the other hand, only after the
4th session we observed a blockade of thermal sensitivity. Regarding cellular changes, we observed a increase of PO expression
after MOB in the ipsilateral nerve when compared to CCI animals.
Conclusions:
In conclusion, we suggest the involvement of PO in rats after the treatment with MOB. Our findings contribute to the
understanding of the mechanisms involved in the therapeutic potential of neural mobilization as a treatment for neuropathic pain.
Keywords: Neural Mobilization, Western blot , chronic constriction injury, hyperalgesia, sciatic
Financial Support: Fapesp, CAPES
Resumo:17-148
EVALUATION OF THE THERAPEUTIC POTENTIAL OF CANNABIDIOL IN A MURINE MODEL OF LPSINDUCED ACUTE LUNG INFLAMMATION.
Almeida, V. I. ; Ribeiro, A. ; Ferraz-de-paula, V. ; Pinheiro, M. L. ; Akamine, A. T. ; Quinteiro-filho, W. ;

Palermo-neto, J. ; Gomes, P. F.
Patologia/Faculdade de Medicina Veterinria e Zootecnia, USP
Objectives:
Endocannabinoid system has become a topic of great interest in pharmacology due to its remarkable distribution in mammal
organisms and capacity to play a modulatory role on diverse physiological functions, including immune system modulation.
Acute lung injury is a murine model of Acute Respiratory Distress Syndrome (ARDS), a condition developed by many patients
with sepsis, which affects thousands of people every year. Experimental data from our laboratory show that CBD has antiinflammatory activity if given previous to the induction of acute lung inflammation. Therefore, the goal of our work was to
evaluate possible therapeutic effect of CBD on ARDS, i.e., given after (6 h) the induction of inflammation.
Twenty-one C57BL/6 mice were used and divided randomly into 3 groups: PBS+vehicle, LPS+vehicle, and LPS+CBD (n = 45/group). Each group received CBD (20mg/kg) or vehicle i.p. 6 hours after nasal instillation of LPS or sterile PBS (1 uL / mouse
g). Twenty-four hours following nasal instillation of LPS or PBS, mice were euthanized and bronchoalveolar lavage (BAL), bone
marrow, spleen and blood were harvested to the analysis of total leukocyte count. It was also harvested a fraction of the lung to
measure myeloperoxidase (MPO) activity. Data were analyzed using ANOVA followed by Tukey-Kramer multiple comparison
test. We showed that LPS+vehicle group increased the total cell count in the BAL (F(2,14)=33.9,p
Conclusions:
We have previously shown that CBD given in a prophylactic manner was able to decrease several inflammatory parameters. Here
we showed that CBD (20mg/kg) given following a therapeutic protocol, i.e., after the induction of lung inflammation, reduce the
acute lung inflammation. Literature data point forwards some possible mechanisms involved in the anti-inflammatory action of
CBD, such as inhibition of NF-kB. In addition, there are reports that anti-inflammatory properties of CBD are triggered by A2A
adenosine receptor. These factor are new being analyzer.
Keywords: Acute lung inflammation, Cannabidiol, Cannabinoids, Lipopolysaccharides, Neuroimunomodulation
Resumo:17-149
ATTENUATION OF ACTIVITY IN AN ENDOGENOUS ANALGESIA CIRCUIT DURING THE CRONIFICATION
PERIOD OF INFLAMMATORY PAIN IN RATS.
Miranda, J. 1; Tambeli, C. H. 2; Lamana, S. M. 1

1
Faculdade de Odontologia de Piracicaba, FOP - UNICAMP
2
Universidade Estadual de Campinas, UNICAMP
Objectives:
The aim of this study was to assess whether persistent mechanical inflammatory hyperalgesia, during its induction period, affects
the analgesia induced by activation of an endogenous pain modulation circuit that can be activated by peripheral nociceptive
stimulation. Once activated, this neural circuit named ascending nociceptive control (ANC) induces analgesia mediated by
endogenous opioids in Nucleus Accumbens. ANC was activated by capsaicin injection in the forepaw, and the nociceptive test
was applied in a distant region, i.e. the hind paw.
Methods: Male Wistar rats (180-220g) were used. Persistent mechanical hyperalgesia was induced by the subcutaneous injection
of PGE2 (100ng/50l) into the hind paw of rats for a period of 14 consecutive days. To verify the progression of mechanical
hyperalgesia, the basal nociceptive threshold was assessed on days 1 and 7 of the induction period of persistent hyperalgesia and
on days 1,7,14, and 21 after discontinuation of the treatment with PGE2. The nociceptive mechanical threshold was evaluated by
the method of compression of the paw of the animal, first described by Randall & Selitto (1957). To verify whether peripheral
noxious stimulation attenuates the hyperalgesic response, the nociceptive threshold was assessed on day 1 of the induction period
of persistent hyperalgesia, 3 hours after the injection of PGE2 and on day 7of the induction period of persistent hyperalgesia, 3
hours after the injection of PGE2. Saline or Cys2,Tyr3,Orn5,Pen7 amide (CTOP), was administered intra-nucleus accumbens 10
min before the intraplantar injection of capsaicin (125g/50l) or vehicle into the forepaw of rats, and the nociceptive threshold
of the animals was assessed 0, 15, 30, 45 and 60 minutes later, in the hind paw. The intra-nucleus accumbens administration was
performed by a cannula stereotaxically positioned. The locomotor activity was evaluated by Rotarod after nociceptive testing. A
two-way repeated-measure ANOVA with one between subjects factor (i.e., treatment) and one within subjects factor (i.e, time)
followed by Tukey test, was used to determine if there were significant (p 0.05) differences in nociceptive mechanical threshold
among the groups. Results: Daily injections of PGE2 induced persistent inflammatory hyperalgesia from day 7 of induction, since
the nociceptive baseline threshold significantly decreased on day 7 (42,5%) of induction and on days 1 (49,5%), 7 (27,8%), 14
(29,5%) and 21 (20,1%) of the period of maintenance of hyperalgesia compared to the basal nociceptive threshold of the 1st day
of the induction period (p
Conclusions:
These findings suggest that inflammatory pain attenuates the activity in endogenous pain control circuits during the period of its
chronification.
Keywords: antinociception, mechanical hyperalgesia, nociceptive stimulation
Resumo:17-150
P-CYMENE ATTENUATES MECHANICAL HYPERNOCICEPTION IN MICE.
Santana, M. F. ; Chaves, D. O. ; Santana, M. T. ; Oliveira, M. G. B. ; Melo, M. S. ; Siqueira, R. S. ;

Cavalcanti, S. C. H. ; Arajo, A. A. S. ; Bonjardim, L. R. ; Jnior, L. J. Q.
Universidade Federal de Sergipe, UFS
Objectives:
This study investigated the effect of p-cymene (PC), a biological monoterpene precursor of carvacrol, on inflammatory
hypernociception induced in mice.
Male Swiss mice were divided into five groups (n = 6, per group), pre-treated with vehicle (saline + tween 80 0.2%), PC (25, 50
or 100 mg/kg, i.p.), 0.5 h after the subplantar injection of 20 l of carrageenan (CG; 300 g/paw), tumor necrosis factor- (TNF; 100 pg/paw), prostaglandine E2 (PGE2; 100 ng/paw) or dopamine (DA; 30 g/paw). Hypernociceptive behavior was measured
with a digital von Frey apparatus at period 0.5, 1, 2, and 3h after the hyperalgesic stimuli. Experimental protocols were approved
by the Animal Care and Use Committee at the UFS (N 26/09). The data were evaluated by one-way analysis of variance
(ANOVA) followed by Tukeys test (p < 0.05). Acute pretreatment with PC, in all doses, inhibited the development of
mechanical hypernociception induced by CG and TNF- (0.5, 1.0, 2.0 and 3.0h; p < 0.001)when compared with the vehicle
group. Additionally, treatment inhibited the hypernociception induced by final mediators of the cascade hypernociceptive, PGE2
(0.5h, in the doses of 25 mg/kg p < 0.05, 50 mg/kg p < 0.001 and 100 mg/kg p < 0.001; 1.0, 2.0 and 3.0h; p < 0.001 in all doses)
and DA (0.5, 1.0, 2.0 and 3.0h; p < 0.001 in all doses)when compared with the vehicle.
Conclusions:
Our results suggest that acute treatment with PC produces anti-hypernociceptive effect in mice probably by a mechanism
dependent on inhibition of cytokines production and/or the final hypernociceptive mediators. Therefore, its tempting to speculate
that PC may represent a new compound for the treatment of painful conditions.
Keywords: HYPERNOCICEPTION, MONOTERPENE, PAIN, p-CYMENE
Financial Support: FAPITEC-SE, CNPq.
Resumo:17-151
HEXANIC AND N-BUTANOLIC FRACTION OF COMBRETUM DUARTEANUM CAMBESS REDUCES
OROFACIAL NOCICEPTIVE BEHAVIOR IN MICE
Melo, M. S. 1; Oliveira, G. F. 1; Souza, T. T. 1; Oliveira, M. G. B. 1; Santana, M. F. 1; Santana, M. T. 1;

Xavier, M. A. 1; Agra, M. F. 2; Tavares, J. F. 2; Quintans-junior, L. J. 1
1
Universidade Federal de Sergipe, UFSE
2
Universidade Federal da Paraiba , UFPB
Objectives:
The purpose of the present study was to evaluate the antinociceptive effect of the hexanic (FH) and n-butanolic fractions (FB) of
Combretum duarteanum Cambess in formalin- and glutamate-induced orofacial nociception on mice.
Male Swiss mice (25-30 g), with 2-3 months of age, were used throughout this study. Mice (n = 8/ per group) were pretreated
with FH or FB at doses of 100, 200 and 400 mg/kg (i.p.), 30 min before of the tests. In all models, nociception was quantified at
those periods by measuring the time (s) that the animals spent face-rubbing in the injected area with its fore- or hindpaws. The
experimental protocols were approved by the Ethics Committee on Animal Research at the Federal University of Sergipe (CEPA:
17/10). The results were analyzed using ANOVA followed by post-Tukey test, were considered significant at p < 0.05. In the
formalin test, was found that the FH reduced orofacial nociception in first phase of the test (48.9%, 60.4%, 47.6%) at doses of
100, 200 and 400 mg/kg and the FB (41.5%, 65.3%) at doses of 100 and 400 mg/kg, p < 0.001. In second phase, FH and FB were
also effective to reduce orofacial nociception (40.0%, 50.4%, 55.3%) and (65.0%, 77.8% and 96.0%, p < 0.001), respectively, at
doses of 100, 200 and 400 mg/kg. In glutamate-induced orofacial nociception, FH and FB produced a significant reduction in
orofacial nociceptive behavior (60.9%, 66.4%, 52.5%) and (63.4%, 66.9%, 90.3%) with p < 0.001, respectively
Conclusions:
Our results suggest that Combretum duarteanum might represent an important tool for treatment of orofacial pain. Further studies
currently in progress will enable us to understand the precise action mechanisms
Keywords: Combretum duarteanum, orofacial pain, formalin, glutamate
Financial Support: CNPq and FAPITEC
Resumo:17-152
ANTINOCICEPTIVE ACTIVITY OF A NEW ALKALOID ISOPROPYL N-METHYLANTHRANILATE FROM THE
ESSENTIAL OIL OF CHOISYA TERNATE KUNTH AND THE ANALOGS METHYL AND PROPYL NMETHYLANTHRANILATE
Pinheiro, M. M. G. 1; Radulovic, N. S. 2; Miltojevic, A. B. 2; Mcdermott, M. 3; Waldren, S. 4; Parnell, J. A.

4
; Fernandes, P. D. 1; Menezes, F. D. S. 3
1
Laboratorio Farmacologia da Inflamaao e do NO/ICB, UFRJ
2
Department of Chemistry/ University of Ni, University Nis
3
Panoz Institute/Trinity College Dublin, TCD
4
Department of Botany/Trinity College Dublin, TCD
Objectives:
Aim:The infusion of leaves of Choisya ternate Kunth (Rutaceae) is popularly employed for their antispasmodic property. The
study the detailed GC-MS analyses of the essential oil of C. ternate revealed the presence of isopropyl N-methylanthranilate
(ternanthranin) (ISOAN) and other volatiles compounds. The essential oil of leaves (EO), ternanthranin and the analogs methyl(MAN) and propyl-N-methylanthranilate (PAN) were assayed for nociception.
Material and methods:EO of C. ternateleaves, collected at Dublin, was obtained by hydrodistillation. The oil and compounds
were analyzed by CG-MS and spectrally characterized (UV-Vis, IR, NMR and MS). Male Swiss mice (2025g; n=6-8) were
orally pre-treated 60 minutes before with EO (at doses 10-100 mg/kg), ISOAN, MAN, or PAN (at doses 0.3-3 mg/kg) and
evaluated in the acetic acid (2%, ip) induced writhing (AA) and hot plate (HP) models. Animal care and research protocols were
in accordance with the principles and guidelines adopted by the Brazilian College of Animal Experimentation (COBEA),
approved by the Ethical Committee for Animal Research/UFRJ (Protocol number, DFBCICB-015). The results are presented as
mean S.D. Statistical significance between groups was determined by ANOVA followed by Bonferronis test (*p
Conclusions:
EO and its minor alkaloid constituents from the leaves of C. ternate produce dose-related antinociceptive action. Furthermore, the
antinociceptive action demonstrated in the present study supports, at least partly, the ethnomedical uses of this plant.
Keywords: anthranilate, nociception, choysia ternate
Financial Support: CAPES, CNPq, FAPERJ.
Resumo:17-153
HEME OXYGENASE 1 INDUCTION REDUCES LUNG OXIDATIVE STRESS AND INFLAMMATION AFTER
INTESTINAL ISCHEMIA AND REPERFUSION INJURY
Bertoni, J. A. 1; Correa-costa, M. 2; Breithaupt-faloppa, A. C. 1; Lino dos Santos Franco, A. 1; Vitoretti, L.

B. 1; Saraiva Cmara, N. O. 2; Tavares-de-lima, W. 1
1
Farmacologia / Universidade de So Paulo, ICB - USP
2
Imunologia/Universidade de So Paulo, ICB - USP
Objectives:
Intestinal ischemia and reperfusion (intestinal I/R) frequently occurs in human pathological conditions, and it has been considered
as a significant cause of Acute Lung Injury (ALI). In your most severe form Acute Respiratory Distress Syndrome (ARDS) is
responsible for significant morbidity and mortality. It is thought that polymorphonuclear neutrophils and mediators generated in
the setting of oxidative stress, such as reactive oxygen species (ROS), have important roles in its pathophysiology. In this sense,
we hypothesize that heme oxygenase-1 (HO-1) up regulation, an enzyme with antiinflamatory and antioxidant activities, could
modulate the expression of antioxidant enzymes and neutrophils recruitment in the lung after an intestinal I/R, therefore inducing
cytoprotection.
Upon anesthesia male Wistar rats (60 days) weighing 200 ~ 300 g were subjected to 45 min occlusion of the superior mesenteric
artery (SMA) and followed by 2 hr of reperfusion. Before ischemia, one group (n=5) of rats had the lymphatic thoracic duct
sectioned and another group of rats (n=5) was treated 48 h and 24 h before induction of SMA occlusion with an HO-1 inductor,
Hemin (10 mg/kg, i.p.). Neutrophils recruitment to the tissues was indirectly measured by myeloperoxidase (MPO) activity. HO1, superoxide dismutase (SOD) 1 and 2, catalase (CAT) mRNA were determined in the lung tissue. Intestinal I/R determined a
significant increase of MPO activity in lung that was reduced in rats upon lymphatic duct sectioned. Hemin treatment of rats
increased the expression of HO-1, SOD-1 and SOD-2, but did not modify the expression the CAT in lung tissue. In addition, the
MPO activity in lung tissue was significantly reduced in hemin-treated group as compared with non treated rats upon intestinal
I/R.
Conclusions:
Our data indicate that HO-1 decreases neutrophils recruitment and upregulates the expression of antioxidant enzymes, reducing
inflammation of lung tissue after intestinal I/R. Moreover lymph factors seem not be involved with HO-1 effects.
Keywords: Acute respiratory distress syndrome, Inflammation, Intestinal ischemia and reperfusion , Heme Oxygenase 1, Lung
Financial Support: FAPESP (2007/07139-3 and 2009/54823-2) / CAPES and CNPq
Resumo:17-154
LOCAL ANTINOCICEPTIVE EFFECT OF CANNABINOIDS IN A RAT MODEL OF PAINFUL DIABETIC

NEUROPAHTY
Schreiber, A. K. 1; Neufeld, M. 1; Enzo, J. 1; Pinto, R. B. 1; Cunha, T. M. 2; Cunha, F. D. Q. 2; Cunha, J. M.

D. 1
1
Farmacologia, UFPR
2
Farmacologia, FMRP - USP
Objectives:
Painful neuropathy is one of the most common secondary complications associated with diabetes. This kind of pain often
responds poorly to standard pain treatments and often a better management is required to treat this condition. Based on reported
antinociceptive properties of cannabinoids, this study was designed to investigate the intraplantar (i.pl.) effect of AM404 (N-(4hydroxyphenyl) arachidonoylethanolamide; an endogenous cannabinoid reuptake inhibitor, or ACEA (arachidonyl-2chloroethylamide), a potent CB1 agonist in model of neuropathic pain induced by streptozotocin (STZ) in rats.
After a 12h-fast, male Wistar rats received one intraperitoneal (i.p.) injection of STZ (50 mg/kg freshly dissolved in citrate buffer,
10 mM, pH 4.5). Hyperglycemia was conrmed 3 days later and at the finish of each study using blood taken by tail prick and a
strip-operated reectance meter. Only rats with a blood glucose concentration higher than 250 mg/dL at the start and end of the
study were included. The formalin test was performed 4 weeks after the onset of hyperglycemia. After 20 min of vehicle or
cannabinoid i.pl. treatments, normoglycemic (Ngl) and diabetic (Dbt) rats (n=6-10 each group) received an ipsilateral injection of
formalin (0.5%, 50 L) in the hind paw, and flinches were counted in 5 min blocks for 1 hour. All procedures were approved by
the Ethics Animal Experiment Committee of Federal University of Paran. Formalin induced a biphasic pattern of inching
responses, from 0 to 10 min (first phase; 1P) and from 15 to 60 min (second phase, 2P). The two phases were separated by a
quiescent period (QP; 10-15 min post injection). As previous described (Eur J Pharmacol 285:189, 1995), at any given
concentration of formalin, Dbt rats show a raised rate of paw inching. In this study, vehicle-treated Dbt rats displayed a nonsignicant trend to increase inching during 1P but exhibited increased response during QP and 2P. Three different doses of
ACEA (10, 30 or 100 ug, in 50 uL) were tested at both Ngl and Dbt rats. In both groups, only the two higher doses affected pain
scores during the 1P of formalin test. In Ngl rats, treatment with ACEA (30 or 100 ug) reduced the frequency of flinches by 45%
and 58%, respectively. In Dbt group, the pain scores were reduced by 58% and 82% after 30 or 100 ug ACEA-treatment,
respectively. Interestingly, the i.pl. treatment with AM 404 (75 ug) promoted significant reduction of all phases of formalin test in
Dbt rats, reaching the inhibition of 33%, 71% and 52%, during 1P, QP and 2P, respectively. Using the same dose of AM404 in
Ngl rats, the chemical hyperalgesia was reduced by 43% and 77% during 1P and 2P, respectively. Additional dose of 50 ug of
AM 404 was used in Dbt animals, leading a decrease of 41%, 51% and 60% in 1P, PQ and 2P, respectively. Neither ACEA nor
AM404 treatments altered plasma glucose levels when compared with control rats.
Conclusions:
The results of this study show that local administration of selective CB1 agonist or endogenous cannabinoid reuptake inhibitor
are effective in ameliorating chemical hyperalgesia in STZ-diabetic rats. Further studies aimed to investigate the involvement of
CB1 and/or CB2 receptors in these responses. Besides, the results suggested that endocannabinoid system could be an interesting
target for new drugs, as seen that an important antinociceptive effect was obtained after very low doses treatment.
Keywords: Diabetic, neuropathy, endocannabinoids, rat, local
Financial Support: CAPES; Fundao Araucria.
Resumo:17-155
POLYSACCHARIDES OF CAESALPINIA FERREA PODS INHIBITS CELL MIGRATION AND PROTEIN

LEAKAGE INDUCED BY CARRAGEENAN
Silva, R. O. 1; Pereira, L. D. P. 1; Marques-domingos, G. F. O. 1; Assreuy, A. M. S. 1; Pereira, M. G. 1,2

1
Instituto Superior de Cincias Biomdicas, ISBC-UECE
2
Faculdade de Educao, Cincias e Letras do Serto Central, FECLESC-UECE
Objectives:
In vivo experimental studies testing total stem extracts of Caesalpinia ferrea showed analgesic and anti-inflammatory activities in
the model of paw edema induced by carrageenan. Further results obtained from our research group demonstrated antiinflammatory effect of a polysaccharide extract from C. ferrea pods, especially on the late phase of carrageenan-induced paw
edema in rats, an effect that was shown to involve prostaglandins and nitric oxide. In this study, it was evaluated the effect of a
polysaccharide isolated from C. ferrea pods in a cellular model of acute inflammation.
Wistar rats (150-250g) were handled following the principles of the Ethics Committee-UECE (N 09204632-0). The
polysaccharide fraction (FIII) was isolated by ion exchange chromatography (DEAE-cellulose). Animals received intravenous
(i.v.) injection of FIII (1mg/kg in 0.1 mL/100 g weight) or 0.9% saline, 30 min before intraperitoneal (i.p) injection of indirect
(carrageenan; 500 mg) or direct (N-formil-methionil-leucil-fenilalanina fMLP; 50 g) chemoatractants. Animals were sacrificed
4 h later and peritoneal fluid collected for total and differential leukocyte counts and dosage protein dosage. Results were
expressed as mean S.E.M. (n=6) and analyzed by ANOVA and Bonferronis test, considering values p
Conclusions:
The polysaccharide isolated from pods of C. ferrea showed anti-inflammatory effect by inhibiting cell migration, acting both
directly and indirectly on neutrophils. This finding supports the popular use of C. ferrea involving inflammatory desordes.
Keywords: Carrageenan, Caesalpinia ferrea, Cell migration, Polysaccharides, Protein leakage
Financial Support: CAPES, CNPq, FUNCAP; Marques- Domingos G.F.O.
Resumo:17-156
KETAMINE INDUCES PERIPHERAL ANTINOCICEPTION IN RAT BY OPIOID AND CANNABINOID SYSTEM
ACTIVATION.
Petrocchi, J. A. 1; Queiroz-junior, C. M. 2; Paiva-lima, P. 1; Caliari, M. V. 2; Di Marzo, V. 3; Duarte, I. D. G.

1
; Romero, T. R. L. 1
1
Laboratrio de Dor e Analgesia, Departamento de Farmacologia, ICB-UFMG
2
Lab. de Patologia Odontolgica, Faculdade de Odontologia, Fac. Odonto-UFMG
3
Inst. of Biomol. Chemistry, Consiglio Nazionale Ricerche, C.N.R.I
Objectives:
Although ketamine shows a peripheral analgesic component, the base of this mechanism is not completely elucidated. Thus the
aim of this study was obtain evidences for the involvement of endogenous opioids peptides and endocannabinoids in the
peripheral antinociceptive effect induced by ketamine.
The rat paw pressure test was used and hyperalgesia was induced by intraplantar injection of prostaglandin E2 (2 g/paw). The
immunohistochemistry and the HPLC chromatography were performed to verify the participation of beta-endorphin and
endocannabinoids, respectively, in the ketamine-effect. All drugs were administered locally into the right hind paw of Wistar
male rats with n > 4 animals per group. The data were statistically analyzed by one-way analysis of variance (ANOVA) and the
post-hoc Bonferroni test for multiple comparisons. Probabilities of less than 5% (P
Conclusions:
The results provide evidences that ketamine probably induces peripheral antinociceptive effect by release of beta-endorphin and
anandamide to activate the selective , delta-opioid receptors and CB1 cannabinoid receptor.
Keywords: Peripheral antinociception, Ketamine, opioid system, Cannabinoid system
Financial Support: CNPq (473758/2007-5) fellowships by CAPES.
Resumo:17-157
OXIDATIVE STRESS IN CHRONIC POST-ISCHEMIA PAIN (CPIP), A MOUSE MODEL OF COMPLEX
REGIONAL PAIN SYNDROME-TYPE I
Bratti, T. 1; Bobinski, F. 1; Martins, D. F. 1; Mazzardo-martins, L. 1; Winkelmann-duarte, E. C. 2; Santos, A.

R. S. 1
1
Depto.C.Fisiolgicas, Universidade Federal de Santa Catarina, UFSC
2
Depto.C.Morfolgicas, Universidade Federal de Santa Catarina, UFSC
Objectives:
AIM: Rats presented microvascular injury after they were submitted to hindpaw ischemia and reperfusion (IR) and developed
chronic post-ischemia pain (CPIP) with mechanical hypersensitivity (Pain Med, 11: 1224, 2010). Here we investigate in mice
whether this hypersensitivity is associated with oxidative stress.
METHODS: 70 male Swiss mice (2535 g, n=14) were divided in 5 groups: (1) CPIP + NAC: submitted to CPIP and treated with
N-acetylcysteine (NAC) (500 mgkg, i.p.); (2) CPIP + V: submitted to CPIP and treated with vehicle (10 ml/kg, i.p.); (3) CPIP:
submitted to CPIP; (4) S: Sham; (5) N: Naive. CPIP induction in mice was adapted from rats (Pain. 112: 94, 2004) and the
experiments were performed after approval of the protocol PP00509 by the Institutional Ethics Committee for Animal Research
(CEUA) of UFSC. Mice were anesthetized with chloral hydrate (7%, 0.6 ml/kg, i.p.) and an elastic O-ring for braces (Elstico
Ligadura 000-1237, Uniden) was placed around the mouses right hindlimb just proximal to the ankle joint producing a tight-t
that induced ischemia and it was left on the limb for 3 h. Sham mice received the same treatment, except that the O-ring was cut
so that it only loosely surrounded the ankle. To establish the potential antihypersensitivity effect of free radical scavengers,
groups 1 and 2 were pre-treated with NAC or vehicle. The mechanical hypersensitivity was measured according the modified up
and down method using von Frey lament both before and at different time-points after treatment (0.5, 1, 2, 4, 6 and 24 hours) at
2 and 7 days after ischemia and reperfusion (IR). Besides, seven days after IR, groups 3, 4 and 5 were killed by decapitation
without anesthesia and both the plantar surface of the hind paw and the lumbar portion of the spinal cord (L1 to L6) were
collected for thiol levels estimation. These tissue was homogenized and then centrifuged. The supernatant was divided into 2
aliquots and one was used to determine total thiol content while the other was used to determine the non-protein thiol content in
the sample. The color of the solution resulting from the reaction was measured using ELISA (R&D systems). Protein thiol
content was calculated by subtracting the values of non-protein thiols from total thiols. Statistic analysis was performed using
one-way ANOVA, with the Student-Newman-Keuls test as the post-hoc test or t test when appropriate. Results are presented as
the mean S.E.M. for each group. p < 0.05 was considered significant. RESULTS: The time course trials of both 2 and 7 days
following hind paw IR demonstrated von Frey thresholds that were signicantly lower than baseline (p < 0.001). Conversely, 30
minutes following treatment with NAC, von Frey thresholds were significantly different from mice treated with vehicle (p <
0.001), suggesting that NAC reversed the mechanical hypersensitivity caused by IR for at least 2 hours. Seven days after IR, mice
had increased total thiol levels, protein thiol levels and nonprotein thiol levels (p < 0.001) in both hind paw and spinal cord when
compared to naive.
Conclusions:
CONCLUSION: The present study demonstrates that NAC suppressed pain behavior and hind paw tourniquet produced an
increase in thiols levels, suggesting a key role of free radicals that might be able to cause damage to the venules and capillaries.
Keywords: N-acetylcysteine , tiols, pain
Financial Support: UFSC and Capes
Resumo:17-158
EFFECTS OF LOW LEVEL LASER (LLL) AND LIGHT EMITTING DIODE (LED) TO THE LOCAL EFFECT
INDUCED BY BOTHROPS MOOJENI SNAKE VENOM (BMJV)
Nadur- Andrade, N. 1; Zamuner, S. R. 1; Barbosa, A. M. 2; Cogo, J. C. 2

1
Cincias da Reabilitaco/ Universidade Nove de Julho, UNINOVE
2
IP&D/ Universidade do Vale do Paraba, UNIVAP
Objectives:
Bothrops snake venoms produce marked local effects including edema, hemorrhage, pain and necrosis. The currently treatment of
Bothrops accidents is the soroterapy. However, this treatment is inefficient in neutralizing the local effects. In the present study
the therapeutic effect of the LLL and LED therapy was evaluated on edema formation and hemorrhage, induced by BmjV.
Furthermore, the effectiveness of antivenom used alone or in combination with LLL or LED treatment was also evaluated.
Male Swiss mice (22-25 g) were used. Edema was measured by plethismography after subplantar injection of BmjV (1
&microg/paw), or saline solution (control) at 15, 30 min, 1, 3 and 6 h. The hemorrhagic activity was evaluated after injection of
20 &microg of BmjV, measuring the diameter of the hemorrhagic area on the inner skin of mice, 2 h after venom injection. The
LLL was used in 685 nm, 30 mW of power, 15s irradiation time and irradiated area of 0,2 cm2 which correspond to a laser dose
of 2,2 J/cm2, and the LEDs in the wave lengths of 945nm and 635 nm, 110 and 120 mW of power, density of energy 4 J/cm2,
irradiated area of 1,2 cm2, 41s and 38s irradiation time for the red LED (LEDr) and infra-red (LEDinf) respectively, with two
applications, at 30 min and 3 h after the venom injection or saline. The antivenom (0.1 mL/5mg; i.v.) was applied i.v 30 min after
venom injection. Ethics Committee: A017/CEP/2008. BmjV induced an edematogenic effect from 30 min up to 6 h with
maximum peak at 1 h, and also caused a significant hemorrhagic activity in the skin of the animals 2 h after venom injection. The
edema caused by the BmjV was reduced by 77, 84 and 70 % by LLL, LEDinf and LEDr, respectively. Antivenom treatment did
not neutralize the edematogenic effect caused by BmjV. The combined treatment with antivenom therapy and LEDs or LLL
significantly reduced the edema at the maximum peak by 55% for LLL, 64% for the LEDinf and 53% for LEDr. The hemorrhage
caused by the BmjV was reduced by 47, 39 and 47% with LLL, LEDinf e LEDr, respectively. The antivenom, administered
through i.v 30 min after venom injection, had a significant effect on the neutralization of hemorrhage halo induced by BmjV,
causing a reduction of 25% of the hemorrhagic halo. Likewise, treatment of antivenom therapy combined with laser and LEDs
was effective to reduce the hemorrhagic halo in the order of 25% for the LLL, 36% for the LEDinf and 27% for LEDr.
Conclusions:
These results highlight the benefit that the LLL and LED offers as an alternative therapy in the treatment of snake accidents
caused by bothropic venom.
Keywords: low level LASER, Light emitting diode, Bothrops moojeni, edema, hemorrhage
Financial Support: FAPESP n 2008/02297-2X, FVE-UNIVAP, CNPq.
Resumo:17-159
ANTINOCICEPTIVE ASSESSMENT OF SNAKE VENOM CAUDISONA DURISSA COLLILINEATA AND
IDENTIFICATION OF THE ACTIVE FRACTION.
Oliveira, S. A. 1,2; Magalhes, M. R. 2; Costa, E. A. 1

1
Laboratrio de Farmacologia de Produtos Naturais, ICB, UFG
2
Laboratrio de Toxinologia,CEPB, PUC-GO
Objectives:
The venoms of snakes of the subfamily Crotalinae have a great diversity of biological activities. The venom of the snake
Caudisona durissa promotes a series of neurologic and myotoxic accidents in humans without causing pain. In contrast victims
feel analgesia at the injection site a few minutes after the accident. Studies show that the snake venom of Caudisona durissa
terrificus has analgesic activity and other activities. In addition to molecules that act on the circulatory system and coagulation,
several other studies are being developed aiming to isolate molecules that may have analgesic effects (Toxicon. 44:1, 2004, Drug
News Perspect.19: 381, 2006) antimicrobial (Biopolymers. 47: 415, 1998; Toxicon. 44:305, 2004; Toxicon. 45:817, 2005;
Toxicon. 45:807, 2005; Toxicon. 48:227, 2006, Biochem J. 402:93, 2007), or anti-inflammatory (Biochem Biophys Res
Commun.302: 193, 2003).Despite the venom in nature be considered unsuitable for therapeutic use there is the possibility of
isolated constituents to be useful for the treatment of different pathologies. The aim of this study was to identify the fraction with
antinociceptive activity of snake venom of Caudisona durissa collilineata.
The crude venom was subjected to fractionation by HPLC with a gradient 2070% acetonitrile with 0.1% TFA at a flow rate of
1.0 mL/min. Fractions of 1.0 mL were collected. The antinociceptive activity was evaluated against a chemical stimulus (writhing
test induced by acetic acid 0.6%, 0.1 mL/10g,) 20 min after i.p. treatement and 45 min after p.o. treatement and the writings
number recorded by 20 min. In this test the pre-treatment with the crude venom of Caudisona durissa collilineata (Cdc-b) at
doses of 20, 40 and 60g/kg, by intraperitoneal route, significantly inhibited the writhing of 40.12 6.24 control group (saline,
ip) to 14.12 5.26, 7.75 2.48 , 13 2.52, respectively. The fraction that exhibited antinociceptive activity, called FrS at
a dose of 40g/Kg i.p, reduced the number of writhing (40.12 6.24) control group (saline, ip) to 11.43 2.46 . The oral
treatment with Cdc-b and 40g/Kg FrS 40g/Kg reduced the number of contortions of 41.44 0.62 (control group) 2.33 to
19.00 and 2.77 0.75 , respectively.

Conclusions:
The present study revealed that the FrS fraction of the venom of South American rattlesnake, Caudisona durissa collilineata
contributes to the antinociceptive effect caused for crude venom by both oral and intraperitoneal treatment. We have also
demonstrated that the effects of FrS fraction, administered orally, should be mediated by activation of opioid receptors. The
antinociceptive properties of FrS, its special quality of being orally active in low doses, and its long-lasting effect emphasize its
therapeutic potential.
Keywords: Caudisona durissa collilineata, active fraction , writings, antinociception
Financial Support: CAPES, CNPq, FAPEG, PUC-GO.
Resumo:17-160
FEMALE SEX HORMONES MODULATE DIFFERENTIAL CELLULAR RECRUITMENT AND TRACHEAL
REACTIVITY IN A MICE MODEL OF LIPOPOLYSACCHARIDE-INDUCED LUNG INFLAMMATION
Gimenes-junior, J. A. 1; Lino-dos-santos-franco, A. 1; Vitoretti, L. B. 1; Ligeiro-de-oliveira, A. P. 2; Moriya,

H. T. 3; Palermo-neto, J. 4; Oliveira-filho, R. M. 1; Vargaftig, B. B. 1; Tavares-de-lima, W. 1
1
Depto. de Farmacologia / Instituto de Cincias Biomdicas I, ICB I / USP
2
Depto. de Imunologia / Instituto de Cincias Biomdias IV, ICB IV / USP
3
Laboratrio de Engenharia Biomdica / Escola Politcnica, LEB / PTC / EPUSP
4
Depto. Patologia / Fac. de Medicina Veterinria e Zootecnia, VPT / FMVZ / USP
Objectives:
Though it is known that acute lung injury (ALI) induced by lipopolysaccharide (LPS) exposure is characterized by cell migration,
edema and altered airway responsiveness, the role of lung inflammation on the induction of altered airway reactivity after
endotoxin exposure remains undefinied. Female sex hormones (FSH) are implicated in innate and adaptative immune responses
and estradiol reportedly protects or deteriorates asthma symptoms. Previous data from this laboratory revealed that ovariectomy
(OVx) blunted LPS-induced lung inflammation in mice after 24 h. Here, we expanded the study and assessed the influence of
FSH on airway smooth muscle reactivity to methacholine (MCh).
7 day-OVx female mice (C57BL/6, n = 5-8/group, 7-10 wk old, mean body weight = 23 g) were subjected to intranasal
instillation of LPS (Escherichia coli, 100 g/ml, 1 l/g) or saline (0.9%, 1 l/g). Mice were euthanized 24 h later and the cells
recovered in bronchoalveolar lavage (BAL). Total cells coutings were made in Neubauer chambers and differential coutings were
made upon optical microscopy. The thacheal reactivity to MCh were determined using a Powerlab system. Otherwise intact
animals subjected to false operation (Sham-OVx) served as controls. Data (mean SEM) were analysed by ANOVA followed by
the Student Newman-Keuls post test (cells BAL) and the Students tailed paired t-test (thacheal reactivity). The GraphPad
Prism version 5.03 was used for this purpose. P
Conclusions:
FSH control lung inflammation by downregulating accumulation of inflammatory cells induced by LPS exposure. Thus, it is
conceivable that FSH-depriving conditions could blunt endogenous control of inflammatory process. The OVx-induced increase
in cellular recruitment after LPS coexisted with a reduced tracheal reactivity, clearly showing that the lung inflammatory
response and the tracheal responsiveness to LPS exposure are under distinct pathways of control by FSH.
Keywords: Lung, Inflammation, Lipopolysaccharide, Female Sex Hormones, Mice
Financial Support: FAPESP (2009/52782-7 and 2009/51886-3).
Resumo:17-161
-TERPINEOL REDUCES NOCICEPTIVE BEHAVIOR IN MICE
Oliveira, M. G. B. D. 1; Santana, M. F. D. 1; Santana, M. T. 1; Santos, A. B. D. 1; Guimaraes, A. G. 1;

Siqueira, J. S. 1; Sousa, D. P. D. 1; Almeida, R. N. D. 2; Bonjardim, L. R. 1; Quintans-jnior, L. J. 1
1
UNIVERSIDADE FEDERAL DE SERGIPE, UFSE
2
Objectives:
-Terpineol (TPN) is a monoterpenoid alcohol present in the essential oils of several species of the Eucalyptus genus
(Myrtaceae). The aim of the present study was investigate the antinociceptive of TPN in rodents.
The antinociceptive effect of TPN was examined using the acetic acid (0.85%) writhing reflex, formalin (20 l of 1%), glutamate
(20 l of 20 mol), and capsaicin (20 l of 1.6 g)-induced nociception tests. Mice divided into five groups (eight per group),
were pretreated with vehicle (saline + tween 80 0.2%) or TPN (25, 50 or 100 mg/kg, i.p.) 0.5 h before experiments. Experimental
protocols were approved by the animal care and use Committee (CEPA/UFS: 26/09) at the Federal University of Sergipe. The
obtained data were evaluated by one-way analysis of variance (ANOVA) followed by Tukey's of test. These results were
presented with Mean SEM, in all cases, differences were considered significant if p < 0.05. TPN produced significant a
analgesic effect by reduction at the early and late phases of paw licking, 1st phase: 25 (46.0 9.2), 50 (49.1 4.9) and 100 mg/kg
(40.5 6.2), 2nd phase: 25 (17.3 8.9), 50 (0.4 0.3) and 100mg/kg (14.1 10.3), with p< 0.01 in the formalin test and reduced
the writhing reflex in mice 25 (4.5 1.3), 50 (1.5 0.7) and 100mg/kg (0.7 0.4) in the writhing test with p< 0.01) nociceptive
protection, 25 (32.5 5.8), 50 (32.1 5.2) and 100mg/kg (33.0 3.4). When the capsaicin-induced nociception test was
conducted, TPN produced dose-related inhibition of the nociceptive behavior, 25 (25.5 4.4), 50 (13.2 2.7) and 100 mg/kg (4.5
1.0) with p< 0.01 or p< 0.001). Such results were unlikely to be provoked by motor abnormality.
Conclusions:
Together, these results suggest that TPN, an important monoterpene of the Eucalyptus species, might represent an important tool
for treatment of pain conditions. Further studies currently in progress will enable us to understand the precise action mechanisms.
Keywords: -terpineol, formalin, glutamate, monoterpene, pain
Financial Support: FAPITEC-SE, CNPq
Resumo:17-162
RELATIONSHIP BETWEEN NIGROSTRIATAL DOPAMINE AND NOCICEPTIVE MODULATION: THALAMIC
HYPERACTIVATION IN HEMIPARKINSONIAN RATS
Domenici, R. A. 1; Maciel, S. T. 1; Fonoff, E. T. 2; Pagano, R. D. L. 1

1
Hospital Sirio-Libanes, IEP- HSL
2
Dep. Neurology, University of Sao Paulo Medical School, USP
Objectives:
Parkinson Disease (PD) is a chronic progressive neurodegenerative disorder characterized by selective loss of nigrostriatal
dopaminergic neurons, modifying the extrapyramidal motor control. Chronic pain is a common complaint in PD, which may
precede the motor symptoms and is commonly neglected and poorly understood. Therefore, it was showed a reduction of
nociceptive threshold in hemiparkinsonian rats in different models of nociception. The aim of this study was to investigate the
mechanical and thermal nociceptive threshold of the rats submitted to a PD model and consequently to evaluate the neuronal
activation pattern of the neurocircuitry involved in this response.
Adult male Wistar rats (200-250 g), deeply anesthetized (tribromoethanol 2,5%), were submitted to a PD model induced by
unilateral striatal stereotaxic injection of the neurotoxin 6-hydroxydopamine (6-OHDA; 12 g/2l), on the left side. Rats injected
with saline or nave were used as control (n=5 animals per group). After 14 days of the lesion, rats were evaluated in the open
field test and no significant change in the motor behavior or stereotyped movements were noted. After 7 and 14 days of the
neurotoxin injection, mechanical and thermal hyperalgesia were investigated by paw pressure and plantar tests, respectively. In
all times evaluated, it was observed a reduction (55%) in the mechanical nociceptive threshold of the hemiparkinsonian rats,
when compare with control group. However, no difference was observed in the thermal response. After 1 hour of the nociceptive
tests, the immunoreactivity (IR) for tyrosine-hydroxylase (TH; dopaminergic cell marker) in the substantia nigra (SN) was
determined. Nigrostriatal damage was characterized by the loss of 62% in TH-IR in cells of the SN, in the injection side, and 21%
in the contralateral side. We also evaluated the IR for Fos protein, a neuronal activation marker, in the periaqueductal gray
(PAG), the SN and the ventral posterior lateral and medial nuclei (VPL/VPM) of the thalamus. Fos-IR increased (30%) in the
VPL/VPM of the lesioned animals, on the injection side, when compared to the pattern observed in the control animals. No
significant change in Fos-IR in the SN or in the PAG was noted.
Conclusions:
These results show that unilateral dopaminergic inhibition induces bilateral mechanical hyperalgesia, suggesting that the
dopamine in the mesostriatal system has an important role in modulating nociceptive process. Thalamic hyperactivation suggests
a close interaction between nigrostriatal dopaminergic system and pain afferent pathway in PD. Further investigation of the
mechanisms involved in these nociceptive modulation may contribute to improve the clinic treatment of persistent pain observed
during Parkinsons disease.
Keywords: 6OHDA, Fos protein, Mechanical hyperalgesia, Parkinson disease, Thalamic nuclei
Financial Support: FAPESP and Hospital Srio-Libans
Resumo:17-163
ANTINOCICEPTIVE EFFECTS OF AN ALKALOID EXTRACTED FROM FRUITS OF OCOTEA PUBERULA.
Montrucchio, D. P. 1; Szyminovicz, M. 2; Bobinski, F. 2; Borges, F. R. M. 2; Santos, A. R. S. 2

1
PPG Farmacologia, UFSM
2
Depto de Cincias Fisiolgicas, UFSC
Objectives:
Ocotea puberula is a native brazilian tree found mainly in the south region, and its barks and leaves has been a target for several
phytochemical and some pharmacological studies. The aim of this study was to investigate the antinociceptive properties of
dicentrine, an alkaloid extracted from O. puberula fruits as the majoritary compound, in animal models of acute and chronic
inflammatory pain.
Ocotea puberula fruits collected in Curitiba, state of Paran, were extracted with ethanol in a soxhlet apparatus for 24 hours,
filtered and concentrated under reduced pressure. The concentrated ethanolic extract was then fractioned with solvents of
increasing polarity: n-hexane, chloroform and ethyl acetate. The chloroform fraction, rich in alkaloids, was then submitted to a
column chromatography, from which it was isolated an alkaloid identified as dicentrine. The antinociceptive properties of
dicentrine were tested in male adult Swiss mice, weighing between 25-35g, in an acute nociception model (formalin) and in a
chronic inflammatory nociception model (CFA). In the formalin model, three groups (n=6) were pretreated with a solution of
dicentrine in doses of 10, 30 and 100 mg/kg by oral route 60 minutes prior to the injection of 20L of a 2,5% formalin solution in
the right hindpaw of mice. The time spent licking the injected paw was counted during the first 5 minutes (neurogenic phase) and
between the 15th and 30th minute (inflammatory phase). Dicentrine was able to diminish the licking time in the inflammatory
phase in a dose-dependent manner, with the best result at the dose of 100mg/kg (MI=6214%). In the chronic model, an
inflammation was inducted by the injection of 20L of CFA 50% in the plantar surface of mices right hindpaw. After 24h, the
mechanical hypersensitivity was evaluated as the withdrawal response to 10 applications of 0,4g von Frey hair, and the thermal
hypersensitivity was evaluated in the hot plate (50C) as the latency for paw withdrawal (cut off 20s). Dicentrine (100 mg/kg,
n=8) administered by oral route was able to diminish the mechanical, but not thermal hypersensitivity, up to 2 hours after
administration, during 14 days of daily treatment. After the last von Frey evaluation, the mice were submitted to the open field
test and showed no effect in the locomotor activity. The weight gain was monitored along the 14 days and there was no difference
between treated and control groups.
Conclusions:
The results suggest that dicentrine has an antinociceptive effect on inflammatory conditions, without compromising the
locomotor activity in animal models.
Keywords: Dicentrine, Nociception, Ocotea puberula
Resumo:17-164
SURVIVAL AND OXIDATIVE STRESS OF ZEBRAFISH LARVAE EXPOSED TO ESCHERICHIA COLI
LIPOPOLYSACCHARIDE
Concatto, S. C. 1,2; Leite, C. E. 1,4,5; Bonan, C. D. 3; Menezes, F. P. 3; Campos, M. M. 1; O. Battastini, A. M.

4,5
; Morrone, F. B. 1,2
1
INSTITUTO DE TOXICOLOGIA , INTOX, PUCRS
2
FACULDADE DE FARMCIA, FFARM, PUCRS
FACULDADE DE BIOCINCIAS, FABIO, PUCRS

DEPARTAMENTO DE BIOQUMICA ICBS, DEPBIOQUIMICA, PUCRS
5
FACULDADE DE MEDICINA, FAMED, UFRGS
Objectives:
Zebrafish (Danio rerio), known in Brazil as Paulistinha, has been used in various stages of drug discovery process. It is a useful
alternative, of great scientific value and lower cost, in comparison to other animal models (eg, rodents, dogs and pigs). The
standardization of a model of inflammation in zebrafish larvae, as well as the characterization of the systems involved in this
process might contribute to the validity and safety of its use as a model for screening new drugs. The aim of this study was to
begin the characterization of inflammation in zebrafish larvae exposed to Lipopolysaccharide (LPS) from Escherichia coli, by
assessment of survival and oxidative stress.
Larvae obtained and selected during the experiments were kept in an incubator and later transferred to Petry plates until use. The
experiments were performed in 6-well plates with up to 13 larvae per well. Zebrafish larvae were exposed to Escherichia coli
LPS at concentrations of 0, 50, 100, 150, 200 and 300 g/ml. The survival was assessed for 72 hours. The oxidative stress was
assessed, 4 hours after LPS exposure, by catalase (CAT), superoxide dismutase (SOD) and malondialdehyde (MDA) assay. The
control group and those exposed to 50 and 100 g/ml showed higher survival rates (higher than 95%). The groups receiving 150,
200 and 300 g/ml showed survival rates of 885%, 575% and 0 %, respectively. The groups treated with 150 g/ml of LPS
showed a significant decrease of CAT activity (249%), allied to SOD and MDA increase (187% and 299%, respectively).
Conclusions:
The survival curve shows that the concentration of 300 g/ml induced 100% of fish death and, oxidative stress was induced by
150 g/ml. Additional studies are being conducted to determine the mechanisms involved in the induction of oxidative stress and
death of animals, but probably these events occurred in consequence of over-activation of defense mechanisms of the zebrafish,
similar to sepsis in mammals.
Keywords: LIPOPOLYSACCHARIDE, OXIDATIVE STRESS, ZEBRAFISH LARVAE
Financial Support: FAPERGS/CNPq
Resumo:17-165
MODULATION OF ENDOTHELIN PRODUCTION BY LIPOXIN A4 SUPPRESSES ARTICULAR INFLAMMATION
Conte, F. P. 1; Junior, O. M. D. L. 1; Junior, W. A. V. 2,3; Cunha, F. Q. 3; Penido, C. 1; Henriques, M. G. M.

O. 1
3
Farmacologia / Universidade de So Paulo, USP
1
Farmacologia / Farmanguinhos-Fundao Oswaldo Cruz, FIOCRUZ
2
Cincias Patolgicas / Universidade Estadual de Londrina, UEL
Objectives:
Endothelins (ETs) are pro-inflammatory peptides involved in pain, fever, edema and cell migration. Rheumatoid arthritis (RA) is
a chronic autoimmune inflammatory disease characterized by joint inflammation, as well as local and systemic increased ET-1
levels. The present study demonstrates the effect of ET antagonism in articular inflammation on zymosan- and collagen-induced
arthritis and also show the modulation of ET-1 levels by lipoxin (LX)A4, an endogenous anti-inflammatory mediator.
Two hours after induction of zymosan (500 g/cav, i.a)-induced arthritis in C57BL/6 mice, ET-1 mRNA expression levels were
increased in synovial extracts, which was followed, at 6 and 24 h, by massive edema formation and neutrophil influx into
inflamed joint. Pre-treatment with bosentan (10 mg/kg; i.v.), a dual pharmacological ET receptor antagonist, significantly
impaired 6 h zymosan-induced leukocyte accumulation and edema formation. ET receptor blockade also decreased zymosaninduced production of TNF- CXCL1 and leukotriene(LT) B4 within 6-24 h. Bosentan (10 mg/kg; i.p.) post-treatment effectively
attenuated clinical score of established collagen-induced arthritis in male DBA/1 mice and caused a significant decrease on paw
edema (30 and 35% of inhibition, respectively). In addition, bosentan post-treatment reduced lymph node weight and cellularity,
with concomitant suppression in the numbers of CD3+CD25+CD71+ cells. LXA4 in vivo pre-treatment (20 ng/cav, i.a.)
significantly impaired zymosan-induced preproET-1 mRNA expression (80 % of inhibition; n=5) in C57BL/6 mice with
concomitant suppression of zymosan-induced edema formation and neutrophil influx (74 and 61% of inhibition, respectively;
n=6). In addition, in vitro pre-treatment of neutrophils with LXA4 (1-100 nM) inhibited ET-1 (100 nM)-induced activation and
chemotaxis, evaluated by shape change and boyden chamber assay, respectively.
Conclusions:
Our results suggest that pharmacological blockade of ET receptors, or the modulation of ET-1 expression by LXA4, may serve as
a possible novel therapeutic tool for the treatment of inflammatory and autoimmune articular diseases.
Keywords: artrite, bosentan, endotelina, lipoxina A4, neutrfilo
Resumo:17-166
NORADRENALINE INJECTION INDUCES PERIPHERAL ANTINOCICEPTION BY ADRENERGIC,
OPIOIDERGIC AND CANNABINOIDERGIC SYSTEM INTERACTION.
Souza, T. C. 1; Di Marzo, V. 2; Romero, T. R. L. 1; Duarte, I. D. G. 1

1
Laboratrio de Dor e Analgesia, Departamento de Farmacologia, ICB-UFMG
2
Inst. of Biomol. Chemistry, Consiglio Nazionale Ricerche., C.N.R.I
Objectives:
Despite the classical peripheral pronociceptive effect of noradrenaline (NA), recently studies showed the involvement of this
endogenous adrenoceptor agonist in intrinsic control of peripheral pain under interaction with the immune system. Thus, the aim
of this study was to verify the opioid and the cannabinoid system participation in the peripheral antinociceptive effect of NA.
The rat paw pressure test was used and hyperalgesia was induced by intraplantar injection of prostaglandin E2 (2 g/paw). All
drugs were administered locally into the right hind paw of Wistar male rats with n > 4 animals per group. The HPLC
chromatography was performed to verify the participation of endocannabinoids in this event. NA (05, 20 and 80 ng/paw) elicited
a local inhibition of hyperalgesia (35%, 55% and 85%, respectively). The non selective alpha2 adrenoceptor antagonist
yohimbine (05, 10 and 20 g/paw), the non selective alpha1 adrenoceptor antagonist prazosin (0.5, 1 and 2 g/paw) and the non
selective beta adrenoceptor antagonist propranolol (150, 300 and 600 ng/paw) blocked (20%, 60% and 100%; 22%, 65% and
100%; 20%, 50% and 100%, respectively) the antinociception effect induced by NA, highest dose. The local effect of NA was
antagonized by the non selective opioid receptor naloxone (25, 50 and 100 g/paw) (35%, 50% and 100%, respectively), as well,
by the selective -opioid receptor antagonist clocinnamox (10, 20 and 40 g/paw) (15%, 40% and 100%, respectively) and by the
selective delta-opioid receptor antagonist naltrindole (15, 30 and 60 g/paw) (20%, 50% and 100%), but not by the selective
kappa-opioid receptor antagonist NOR BNI (100 g/paw). In addition, the enkephalinase inhibitor bestatin potentiated the
antinociceptive effect of a low dose of NA from 60% to 90%. The CB1 cannabinoid selective antagonist AM251 (20, 40 and 80
g/paw) (30%, 55% and 100%, respectively) but not the CB2 cannabinoid selective antagonist AM630 (100 g/paw) blocked the
noradrenaline-effect. The anandamide amidase inhibitor MAFP (2 g/paw) and the anandamide reuptake inhibitor VDM11 (20
g/paw) increased from 50% to 90% and from 52% to 95%, respectively, the peripheral antinociceptive effect of the NA low
dose. In additional, the dosage of endocannabinoids (AEA, 2-AG, PEA and OEA) indicated that noradrenaline induces a selective
anandamide (AEA) release in the peripheral site with an increase twice in the lipid extract dosage.
Conclusions:
The results provide evidences that NA probably induces peripheral antinociceptive effect by activation of adrenoceptors in the
resident immune cells to release beta-endorphine or anandamide that activate selectively , delta-opioid receptors or CB1
cannabinoide receptor in peripheral site.
Keywords: Peripheral antinociception, Noradrenaline, Adrenergic system, Opioidergic system, Cannabinoidergic system
Financial Support: CNPq and fellowships from CAPES.
Resumo:17-167
DUAL EFFECT OF INTRAPLANTAR INJECTION OF PERTUSSIS TOXIN IN NOCICEPTION.
Poloni, R. ; Schivo, I. R. ; Cunha, T. M. ; Ferreira, S. H.

Farmacologia/Faculdade de Medicina de Ribeiro Preto, FMRP-USP
Objectives:
Pertussis toxin (PTX) is widely used as a pharmacological tool for inhibiting Gi and G0 proteins, e.g., in studies of the
mechanism of action of antinociceptive drugs. Intrathecal PTX induces hypernociceptive signals compared to neuropathic
syndromes in rats and mice. However, there is a study wherein intraplantar (i.pl.) PTX prevents prostaglandin E2 (PGE2)-evoked
mechanical hypernociception in rats. Due to this contradiction found in literature, the present study evaluated the i.pl. PTX effects
(0.01 - 1 g.paw-1) in nociception and inflammation, as well as i.pl. PGE2-evoked hypernociception (100 ng.paw-1).
Nociceptive evaluation was made through constant pressure and increasing pressure apparatuses on the animals paw (modified
Randall-Selitto and electronic von Frey, respectively). The variation in paw volume was measured by using an
hydroplethismometer. The number of neutrophils was indirectly checked by a colorimetric assay which evaluated the
myeloperoxidase activity. The cytokine concentration was measured by using ELISA. Low doses of i.pl. PTX (0.1 - 0.15 g.paw1) blocks the PGE2-evoked hypernociception in rats, which was confirmed in mice (0.03 - 0.1 g.paw-1). The i.pl. pretreatment
with selective inhibitors of nitric oxide (NO) synthase (l-NOARG), guanylyl cyclase (ODQ), protein kinase G (KT5823) or ATPsensitive K+ channel blocker (glybenclamide) prevented the antinociceptive effect caused by i.pl. PTX in PGE2-evoked
mechanical hypernociception, confirming the data previously revealed by our group. However, if the PTX dose is increased (0.3 -
1 g.paw-1), the rats present mechanical hypernociception, confirmed in mice (0,6 - 1 g.paw-1), and edema, which depends on
the neutrophil migration and increased pro-inflammatory cytokines synthesis/release to the injection situ. Such effects were
partially prevented by the pretreatment with intravenous fucoidin or intraplantar indomethacin, atenolol or dexamethasone. Posttreatment with i.pl. morphine prevented the PTX-evoked mechanical hypernociception. Further, pretreatment with adenylate
cyclase, protein kinase (PK) A or PKC inhibitors also prevented PTX-caused mechanical hypernociception. Mice which are
mutant for toll-like 4 receptors (TLR4) and knockout for MyD88 adaptor molecule did not present either i.pl. PTX-evoked
mechanical hypernociception or neutrophil migration, when compared to the respective wild animals.
Conclusions:
The present study showed that i.pl. PTX at low doses, through the activation of the Arginine/NO/PKG/ATP-sensitive potassium
channel pathway, blocked the PGE2-induced mechanical hypernociception. However, in higher doses, i.pl. PTX is capable of
provoking an inflammatory response, with induction of the mechanical hypernociception, edema, neutrophil migration and
inflammatory mediators synthesis/release. The mechanical hypernociception seems to depend on the signalization pathway which
involves adenylil cyclase / protein kinase (PK) A and PKC. The mechanical hypernociception and the neutrophil migration
evoked by the i.pl. PTX depend on toll-like 4 receptors, as well as MyD88 molecules.
Keywords: Pertussis toxin, Hypernociception, Antinociception, Cytokines, TLR4
Resumo:17-168
EHRLICH ASCITIC TUMOR (EAT): STANDARDIZATION OF THE MODEL 1MACHADO, L.S.,1ZARDO,
R.S.,1FERNANDES, P.D. 1LABORATRIO DE FARMACOLOGIA DA INFLAMAO E DO XIDO NTRICO,
ICB, CCS, UFRJ, RIO DE JANEIRO, RJ.
Machado, L. D. S. ; Zardo, R. S. ; Fernandes, P. D.

FARMACOLOGIA/FARMACIA, UFRJ
Objectives:
Ehrlich tumor is a spontaneous murine mammary adenocarcinoma with rapidly growing and aggressive behavior. Following,
intraperitoneal inoculation of Ehrlich tumor cells, the ascitic volume and number of tumor cells increase progressively. Ascitis is
probably formed in consequence of tumor-induced inflammation, due to the increase in peritoneal vascular permeability. The aim
of this study was standardizes the Ehrlich ascitic tumor (EAT) model to be used as a tool for anti-cancer drug discovery.
0.5 x 106 EAT cells were intraperitonealy injected into Swiss mice (20-25g, n=5-6, ethical committee license #ICBDFBC-015).
Every two days a group of mice were sacrificed, samples of blood and bone marrow lavage were collected for the determination
of total leukocytes count. Ascitic fluid was collected for the determination of volume, total leukocytes count and dosages of
cytokines, nitric oxide (NO), and protein. Results are expressed as media DP. In days 1th, 2sd, 4th, 6th, 8th, 10th,12th,14th
days after tumor inoculation it could be observed an increase in ascitic volume (1.80.1; 1.80.2; 2,10.15; 1.90.4; 2.70.3;
6.31.4; 9.12.6; 15.22.3 mL, respectively), in tumor cells count (0.20.08; 0.30.06; 0.40.07; 1.50.04; 4.20.6; 5.21.1;
5.10.2; 5.20.8 x107 cell/mL, respectively), in animals weight (0.10.05; 0.30.2; 0.80.2; 1.60.9; 3.81.7; 6.11.9; 7.61.4;
10.61.3 g, respectively). It was also observed a significant increase in total leukocytes count in bone marrow (3.81.1; 3.91.2;
4.00.3; 4.61.0; 4.90.9; 3.20.6; 3.60.5; 4.40.6 x106 cells/mL) and blood (2.70.4; 2.70.2; 2,30.07; 2.80.5; 2.40.2;
2.40.2; 3.60.5; 4.40.6 x106 cell/mL). A significant increase in protein extravased (6.41.5; 5.82.6; 40.94.6; 72.82.7;
141.746.0; 281.133.4; 386.629.8; 429.427.1 g/mL), in cytokines (TNF: 0.30.05; 0.30.08; 0.310.02; 0.360.06;
0.570.09; 1.00.2; 1.20.2; 1.30.2 pg/mL and IL-6: 0.60.09; 0.650.09; 0.770.1; 0.80.2; 1.30.4; 2.00.45; 2.00.42;
2.10.4 pg/mL) and NO (6.64.2; 8.33.3; 32.88.1; 69.613.3; 92.59.5; 150.411.7; 192.410.4; 241.536.4 M) were also
observed. After the 10th day an intense hemorrhage was observed and animals died between 15th and 16th days.
Conclusions:
we could observe an increased of ascitic fluid together with tumor cells growth. This phenomenon was accompanied by an
inflammatory response with increase in number of leukocytes, protein extravased, pro-inflammatory cytokines, and nitric oxide.
In conclusion, the EAT model can be used for studies of new anti-inflammatory and anticancer drugs, with treatment during 10
consecutive days.
Keywords: EHRLICH TUMOR, STANDARDIZATION , CANCER
Resumo:17-169
PLATELET ACTIVITY PROFILE IN A MURINE MODEL OF ALLERGIC ASTHMA
Baldissera Jr, L. ; Calixto, M. C. ; Mendes, C. B. ; Antunes, E.

Departamento de Farmacologia/Faculdade de Cincias Mdicas, UNICAMP
Objectives:
Introduction: Platelet activation has been reported in a variety of inflammatory diseases, including bronchial asthma (Clin Exp
Allergy. 36:399-401, 2006). In asthma, platelets have been found to actively participate in most of its main features, including
bronchoconstriction, airway inflammation and airway remodeling (Platelets. 18:319-28, 2007). Obesity is associated with
increased atherothrombotic morbi-mortality risk, and is associated with deterioration in asthma outcomes. However, little is
known about the mechanisms by which platelets are activated in allergic diseases, and whether obesity contributes to the platelet
activation in allergic conditions. Aim: In this study we have examined the in vitro platelet adhesion in lean and obese mice
previously sensitized and challenged with ovalbumin-(OVA).
Methods: Male C57BL/6 mice (15 g) were fed for 10 weeks with standard chow (lean group) or high-fat diet (obese group). On
the eighth week, animals were actively sensitized twice with ovalbumin (100 g of OVA, s.c.) on days 0 and 7. At days 14 and
15, mice were intranasally challenge with OVA (10 mu;g) twice a day (6 h between challenges). Control group were challenged
with saline (50 L). In separate groups, lean and obese mice were treated with anti-TNF- antibody (2 mg/kg, i.p) given 30 min
before first challenges. Venous blood was collected in 3.8% sodium citrate (1:9 v/v) 48 h after OVA challenge, and centrifuged at
100 g, 20C for 15 min. Platelet-rich plasma (PRP) was centrifuged at 800 g, 20C for 13 min. Isolated platelets were then
resuspended in Krebs-Ringer solution (1 mM CaCl2). The adhesion assays (1.2x108 platelets/mL) were carried out in 96-well
plates coated with fibrinogen solution (50 g/ml). Platelet adhesion were induced with ADP (10 M) or thrombin (100 mU/mL).
Results: No significant differences were observed between spontaneous and stimulated-platelet adhesion between asthmatic and
non asthmatic groups. ADP-induced platelet adhesion was significantly lower in obese asthmatic (17.85.0%; p<0.05).
Conclusions:
Conclusion: Our data show that obesity decreases the stimulated platelet adhesion to immobilized fibrinogen at 48 h post OVAchallenge by mechanisms involving TNF- inhibition.
Keywords: asthma, adhesion, obesity, platelet, thrombin
Resumo:17-170
EFFECT OF MEDIUM CHAIN TRIGLYCERIDES, LINOLEIC ACID, SOY LECITHIN AND VITAMINS A ON
WOUND HEALING IN WISTAR RATS.
Magalhes, M. S. F. 1; Macedo, R. N. 2; Monteiro, D. L. S. 2; Oliveira, C. C. 2; Moraes, R. A. 1;

Nascimento, D. F. D. 1; Linhares, J. H. 2; Linhares, A. E. M. S. 2; Moraes, M. E. A. 1; Moraes, M. O. 1
1
Depto. Fisiologia e Farmacologia, UNIFAC-UFC
2
Faculdade de Medicina, UFC
3
Depto. de Cirurgia, UFC
Objectives:
The wound can be defined as any alteration in the anatomic integrity of the skin, resulting from any type of trauma, where it can
even be classified as intentional (surgical incisions) or accidental. The aim of this study was to determine the effects of a
combination of medium chain triglycerides, linoleic acid, soy lecithin and vitamins A and E, in a cutaneous wound model,
considering the cellular events involved in the cascade of wound healing.
A total of 45 young Wistar rats were used, in which a 4 cm2 full thickness skin segment was removed. The rats were distributed
randomly into three groups of 15 animals, Control, Reference and Triglyceride groups, which were treated topically with 0.9%
NaCl, clostebol + neomycin sulfate and the test formulation, respectively, during 12 days. Histological sections were stained with
hematoxylin and eosin, toluidine blue and Massons trichrome. The healing process was assessed using the criteria of Meyers and
special software which quantified mastocytes, collagen fibers and neovessels. The collagen density measured in the Triglyceride
group was significantly higher (P<0.01).
Conclusions:
Therefore, these findings suggest that the test formulation enhances the process of tissue repair.
Keywords: Wound Healing, Medium Chain Triglycerides, Linoleic Acid, SOY LECITHIN, VITAMINS
Financial Support: CNPq, CAPES, FUNCAP, FINEP, MS-RNPC-UNIFAC-HM, and Instituto Claude
Bernard.
Resumo:17-171
SPLENECTOMY INCREASES NEUTROPHIL MIGRATION BUT DOES NOT AFFECT MORTALITY IN LETHAL
SEPSIS
Kanashiro, A. 1; Ferreira, A. E. 1,2; Figueiredo, J. G. 1; Souto, F. O. 1; Alves-filho, J. C. 1; Cunha, T. M. 1;

Cunha, F. Q. 1
1
Farmacologia / Faculdade de Medicina de Ribeiro Preto, FMRP
2
Faculdade de Cincias Farmacuticas de Ribeiro Preto, FCFRP
Objectives:
Sepsis has been considered the leading cause of death in intensive care units. During severe sepsis, a marked failure of neutrophil
migration leading to infectious focus has been observed and is associated with the spreading of infection, resulting in a high rate
of mortality1. This is due to the fact that neutrophils are responsible for controlling the spread of microorganisms. Several
strategies have been investigated to improve this high sepsis mortality associated with the failure of neutrophil migration.
Currently, the role of the spleen in sepsis is contradictory. Recently, it has been demonstrated that splenectomy changes
neutrophil recruitment during a localized inflammatory process2. So, the present study aimed to investigate the role of
splenectomy in polymicrobial sepsis induced by cecal ligation and puncture (CLP) focusing on a study of functionalneutrophil
alterations.
This study was approved by the Ethics Committee of the School of Medicine of Ribeiro Preto - USP, (protocol n. 041/2011).
Spleens were removed from anesthetized mice (Balb/c, male, 22-26 g) and sham animals underwent laparotomy without
splenectomy. Ten days after splenectomy, mice were subjected to CLP surgery under anesthesia. After 6 hours into the neutrophil
migration experiment, the peritoneal cavity was washed with 3 mL of normal saline containing EDTA. Total leukocyte counts
were performed in a Neubauer Chamber while differential leukocyte counts were performed in cytospin preparations. For the
survival study, the animals were monitored daily for 1 wk. Finally, chemotaxis (using Boynder Chamber counts) and killing
(evaluated by bacteria killing function) properties as well as CXCR2 and CD11b receptors expression (evaluated by flow
cytometry analysis) were studied in peripheral neutrophils isolated from splenectomized and sham mice. In the present study,
although an improvement in the failure of neutrophil migration was observed after the development of intra-abdominal sepsis, no
survival benefit was shown for splenectomized animals. Neutrophil properties as well as receptor expression of neutrophils
isolated from splenectomized animals were not altered when compared with the sham operated animals.
Conclusions:
Under our experimental conditions, it was demonstrated that the migration neutrophil improvement after splenectomy surgery did
improve neither sepsis survival nor altered neutrophil functions. Other humoral or cellular immune component(s) present in the
spleen, which may be related to infection control during polimicrobial sepsis, are under investigation.
Keywords: Neutrophil migration, Sepsis, Spleen, Splenectomy, Survival
Financial Support: Santander, CNPq, FAPESP.
Resumo:17-172
A RAPID AND RELIABLE MODEL OF ORAL OEDEMA TO SCREEN FOR NEW ANTI-INFLAMMATORY
DRUGS.
Ortolani, P. L. 1; Reis, W. G. P. D. 1; Bakhle, Y. S. 2; Francischi, J. N. D. 1

1
Depto. Farmacologia, Inst. de Cincias Biolgicas, UFMG
Faculty of Medicine, National Health and Lung Institute, Imperial College
Objectives:
The present study aimed to develop a model to screen for anti-inflammatory effects in the oral cavity.
A range of doses (25 - 1000 mcg in 0.1 ml) of carrageenan (CG) were injected into the upper lips of anesthetized animals
(mixture of ketamine/xylazine,15/90 mg/kg, subcutaneous), or intraplantarly (ipl) in Holtzman rats (150-180 g, male) at time
zero. Control rats (C) received the same volume locally (0.1 ml), either in the lip or the hind paw of physiological saline at time
zero. Increased thickness of the lip or hind paw was measured with a digital caliper (Mytutoyo, Japan) for up to 6 h following
injections. CG induced a significant and dose-dependent increase in lip thickness in comparison with controls (Delta CG at 500
mcg/site= 4.12 0.05 mm; Delta C= 0.023 0.005 mm with a maximal effect occurring at 1 h after injection, in contrast to 3 h
(Delta CG at 500 mcg/site =1.68 0.04mm; Delta C= 0.08 0.004 mm) in rat paws. At these times, non-selective (2 mg/kg
indomethacin; 20 mg/kg ibuprofen), selective (12 or 30 mg/kg celecoxib) non-steroidal anti-inflammatory drugs (NSAIDs) or
dexamethasone (1 mg/kg), given s.c. 30 min before CG, reduced oral and paw oedema to a similar extent.
Conclusions:
Although hind paw oedema is highly predictive of the clinical efficacy of NSAIDs (Inflamm. Res. 45; 531-540, 1996), our results
would suggest that oral oedema in rats could be as useful a model and would take less time in screening for new antiinflammatory drugs.
Keywords: anti-inflammatory, carrageenan, inflammation, oedema, oral cavity
Financial Support: CNPq, FAPEMIG.
Resumo:17-173
HEME MODULATES GASTRIC AND INTESTINAL EPITHELIAL CELLS ACTIVATION
Barcellos-de-souza, P. 1; Moraes, J. A. 1; de Freitas Junior, J. C. 2; Morgado-daz, J. A. 2; Nasciutti, L. E. 3;

Barja-fidalgo, C. 1; Arruda, M. A. 1,4
1
Departamento de Farmacologia e Psicobiologia/IBRAG, UERJ
2
Diviso de Biologia Celular, INCA
3
Instituto de Cincias Biomdicas, UFRJ
4
Farmanguinhos, FIOCRUZ
Objectives:
Gastrointestinal epithelium (GE) works as an intrinsic barrier against microbial invaders. However, the role of GE in immunity is
beyond physical intervention, since it is recognized that GE is able to regulate cellular mechanisms that can distinguish
potentially pathogenic microorganisms from endogenous bacterial flora, as well as coordinate the proper biological response
against them. In many pathological situations arising from chronic inflammatory conditions such as gastritis and inflammatory
bowel disease, GE epithelial cells are deprived of the protection of the mucus secreted by GE-specialized cells. In these
circumstances, notably in gastric and duodenal ulcers, disruption of blood vessels and subsequent lysis of erythrocytes are
common. This may lead to the release of high amounts of heme, which interacts with GE. Previous works from our group have
shown that heme itself is a proinflammatory molecule, activating a number of phlogistic signaling events in a nicotinamide
adenine dinucleotide phosphate oxidase (NADPHox)-dependent manner. In this study we aim to evaluate the effects of heme
upon GE epithelial cells.
A well established non-transformed rat small intestine epithelial cell lineage (IEC6) and a human gastric epithelial cell lineage
(HGE3) were used in this study. Intracellular reactive oxygen species (ROS) generation was measured using a cell-permeable,
oxidation-sensitive dye CM-H2DCFDA (10 M). Cell proliferation was evaluated by tritiated thymidine (10 M) incorporation
to cell DNA. Wound healing assay was performed by scratching confluent cultures and then the number of cells that migrated to
the injured area was counted. Total cell extracts were obtained for immunoblotting. Our results show that free heme, in
concentrations easily found at hemorrhagic sites (about 20 M), evokes intracellular ROS production by IEC6 and HGE3 cells,
which is inhibited when cells are pretreated with diphenyleneiodonium (DPI, 10 M), a NADPHox inhibitor. FAK
phosphorylation, which is related to several cellular responses, is increased by heme (for up to 2 hours) in a NADPHox activity
dependent manner. Heme, in NADPHox-activating concentrations, is involved with IEC6 and HGE3 proliferation and wound
healing. The expression of Heme Oxygenase-1, an enzyme which is involved in heme degradation and is induced during
inflammatory processes, is up-regulated by heme in a NADPHox-dependent manner. Heme effects on transepithelial electrical
resistance (TEER) and on modulating other proinflammatory molecules expression are under investigation.
Conclusions:
These data indicate a prominent role for heme-derived signaling in the pathophysiology of gastrointestinal mucosa dysfunction.
Keywords: Epithelial cells, Heme, NADPH oxidase, Reactive oxygen species
Financial Support: CNPq, FAPERJ, SR-2/UERJ, ABC/UNESCO/LOreal.
Resumo:17-174
JUVENILE IMMUNE CHALLENGE HYPERNOCICEPTION ARE NOT PREVENTED BY ANTINFLAMMATORY
TREATEMANT.
Ferreira, M. S. ; Giusti-paiva, A. ; Nascimento, C. G. O.

INSTITUTO DE CIENCIAS BIOLGICAS/UNIFAL, ICB
Objectives:
Aim: Adult hyperalgesia has some involvement with inflammatory and expressive nociceptive neonatal or juvenile experiences.
In a similar way, lipopolysaccharide (LPS) induced sickness syndrome in neonatal and juvenile subjects can also be responsable
for nociceptive and inflammatory adult response, leading to hypernociception in thouse adult animals. Thus, the aim of this study
was to evaluate the possible adult antihyperalgesic effect of nonsteroidal anti-inflammatory drugs therapy in juvenile rats.
Methods and results: Twenty one days male Wistar rats received one intraperitonial LPS injection (100 g/Kg). Some of those
LPS treated animal received three days of anti-inflammatory analgesic therapy with dipyrone, nimesulide or indometacin. After
become adult (75 days after birth), the mechanical sensibility was measured by electronic von Frey apparatus, where alodinia
could be observed. The results show that IP LPS juvenile treatment induced hypernociception in adult rats (39,63,7 to 82,86,7).
Unlike the presumptions, all the three nonsteroidal anti-inflammatory therapy could not prevent the adult hypernociception. In
fact those animals demonstrated more mechanical sensitivity and alodinia then the only LPS treated group (77,256,25 to
34,44,5) relative to dipyrone. The results were presented as mean and standard error. Analyzed by ANOA and Tukey test.
Conclusions:
Conclusion: The systemic LPS treatment in young rats is able to lead to hypernociception by decreasing the nociceptive threshold
to mechanical stimulus in adult animals. It is important to emphasize that all therapies with nonsteroidal anti-inflammatory drugs
led to a bigger sensitivity to mechanical nociception, and there is probably no influence of those treatments on adult inflammatory
induced hypernociception.
Keywords: ANTINFLAMMATORY, ALODINIA, HYPERNOCICEPTION
Resumo:17-175
ANTI-INFLAMMATORY EFFECT OF NEW TYROSINE KINASES INHIBITORS PLANNED FROM IMATINIB.
Zardo, R. S. 1; Sampaio, T. S. 2; Gonalves, M. R. 1; Lima, L. M. 2; Barreiro, E. J. 2; Fernandes, P. D. 1

1
Instituto de Cincias Biomdicas, ICB-UFRJ
2
Faculdade de Farmcia, LASSBio-UFRJ
Objectives:
The aim of this study was to evaluate the anti-inflammatory effect of new drugs planned from imatinib structure, first tyrosine
kinase inhibitor (TKI) approved for the treatment of cancer. These analogues were designed for a dual action: anti-tumor and
anti-inflammatory, from the implementation of strategies for molecular modification characteristics of medicinal chemistry, as
regioisomers, retroisosterism, simplification, and molecular bioisosterism in classic rings to obtain the target compounds.
The use of animals in this work was approved by the ethical committee of animal experimentation from Centro de Cincias da
Sade (UFRJ), and received the number ICBDFBC-015. Male Swiss mice (20-25g, n=5-7) were used in the formalin-induced
licking response (2.5%, intraplantar) and in the carrageenan (1%)-induced inflammation in the subcutaneous air pouch (SAP)
model. Animals received oral administration of LASSBio1597, LASSBio1598, or LASSBio1599 (30 mg/kg), 1h before formalin
injection. In SAP these drugs were administrated at 1 and 10 mg/kg (LASSBio1597, LASSBio1598 and LASSBio1599), 1h
before and 23h after carrageenan injection. Statistical analyses was performed by ANOVA followed Bonferronis post test (*p
Conclusions:
These results indicate that LASSBio1587, LASSBio1598, and LASSBio1599 have significantly anti-inflammatory action.
Keywords: anti-inflammatory, formalin, air-pouch, tyrosine kinases inhibitors, imatinib
Financial Support: INCT-INOFAR, CNPq, and FAPERJ.
Resumo:17-176
ANTI-INFLAMMATORY PROPERTIES OF THYMOL
Marinho, R. R. ; Santos, J. S. ; Camargo, E. A. ; Thomazzi, S. M.

Departamento de fisiologia/ Universidade Federal de Sergipe, UFS
Objectives:
Thymol is a monoterpene phenol derivative of cymene, found in oil of plants such as Thymus vulgaris, Lippia gracilis, Lippia
sidoides, and Origanum vulgare. It exhibits multiple biological activities, such as antibacterial, antifungal, anti-inflammatory, and
also possesses antioxidant, free radical scavenging, and antilipid peroxidative properties. Previous study shown that thymol
possesses significant inhibitory activity against at least one COX form at concentrations comparable to the active one of
indomethacin and suggest that this agent should be further studied for possible use as non-steroidal anti-inflammatory drugs
(Planta Med. 71:739, 2005). In order to evaluate the actions of this compound, studies were performed on anti-inflammatory
activity.
Methods: Male Swiss mice (20-30 g) and Wistar rats (120-180 g) were obtained from the Central Biotery of the Federal
University of Sergipe and complied with the guidelines on animal care of the Ethics Committee for Animal Use in Research
(CEPA/UFS 55/10). The animals were pre-treated with thymol (10, 30, or 100 mg/kg), dexamethasone (2 mg/kg) or vehicle (n =
6/group). The anti-inflammatory activity was studied using the paw edema model induced by 1% carrageenan (100 L/paw) and
the volume of the paw was measuread at the time 0 and the intervals of 1, 2, 3, and 4 h. The leukocyte migration was induced by
injection of carrageenan (1%, 250 L, i.p.) into the peritoneal cavity of mice and 4 h after carrageenan injection the total cells
were counted. Results: Oral treatment with the thymol (60 min before of stimulation) did not show any significant alteration on
paw edema model, but was capable of reducing the carrageenan-induced MPO activity at 100 mg/kg (5.4 1.87 and 14.26 1.05
UMPO/mg tissue for thymol and vehicle, respectively, p
Conclusions:
Thymol shows anti-inflammatory activity in rodents.
Keywords: anti-inflammatory , Inflammation, paw edema model , leukocyte migration , Thymol
Financial Support: Capes
Resumo:17-177
EXPRESSION OF HO-1 AND COX-2 IN NYLON THREAD LIGATURE-INDUCED PERIODONTITIS IN RATS.
Oliveira, J. M. 1; Souza, R. B. 1; Chaves, H. V. 1; Vieira, A. M. 1; Ribeiro, K. A. 2; Cunha, R. M. S. 2; Silva,

A. A. R. 1; Pereira, K. M. A. 1; Pinto, V. P. T. 1; Bezerra, M. M. 1
1
Universidade Estadual Vale do Acara, UVA
Objectives:
Periodontitis is an inflammatory disease characterized by alveolar bone resorption. During inflammatory response heme
oxygenase-1/ biliverdin/carbon monoxide (HO-1/BVD/CO) and cyclooxygenase-2 (COX-2) pathways are activated. The aim of
this study was to analyse both COX-2 and HO-1 mRNA expression during periodontitis in rats.
Wistar rats received a nylon thread ligature around the molars and sacrificed after 3, 7, 11 or 14 days. Alveolar bone loss (ABL)
was measured using the ImageJ software or histopathologic analysis. Quantitative Real Time PCR (qRT-PCR) for COX-2 and
HO-1 mRNA expression in gingival tissues was perfomed using Mastercycler ep realplex4 (Eppendorf) and Power SYBR
Green Master Mix (Applied Biosystems). Primers were designed by PrimerBlast using a GenBank (NCBI) Reference Sequence
of COX-2 (NM_017232.3) and HO-1 (NM_012580.2) to Rattus norvegicus. The efficiency amplification for all genes was
verified. The Ct method was used to calculate COX-2 and HO-1 mRNA relative expression levels. Expression level of the
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was using as the endogenous control (housekeeping). Data are
shown as meanSEM. Significant (P
Conclusions:
The present data suggest that both HO-1/BVD/CO and cyclooxygenase-2 (COX-2) pathways could be involved in the
progression of periodontitis.
Keywords: Alveolar bone, Cyclooxygenase-2 , Heme oxygenase-1, mRNA, Periodontitis
Financial Support: Funcap; CNPq
Resumo:17-178
ALLERGIC INFLAMMATION MODIFIES THE PHENOTYPE AND FUNCTIONALITY OF LUNG FIBROBLASTS
IN 3D-CULTURE
Dalzy, D. V. ; Guimares-silva, A. M. ; Garzoni, L. R. ; Perez, S. A. C. ; Silva, P. M. R. ; Martins, M. A.

FUNDAO OSWALDO CRUZ, FIOCRUZ
Objectives:
Subepithelial fibrosis is a prominent feature of lung remodeling in asthma of all severities. The phenomenon strongly correlates
with decline in lung function, but its mechanistic basis remains poorly understood. Since lung fibroblasts are major structural
cells orchestrating fibrotic remodeling in asthma, we have here studied and compared the morphological and functional changes
of lung fibroblasts, from normal and asthmatic mice, cultured in a three-dimensional organized cellular arrangement
(spheroids). The central idea was to have lung fibroblast spheroids as an alternative system to study lung tissue remodeling in
vitro.
Sensitized Balb/C mice were challenged with ovalbumin or saline three times a week for two consecutive weeks, starting 30 days
post-sensitization. 24 h after last provocation, digested lung cells were cultured, and homogenous lung fibroblasts (obtained in
three passages) were eventually plated on agarose coated 96-well plastic dishes, without/with IL-13 (10-80 ng/ml). Subepithelium
fibrosis, leukocyte infiltration, cytokine release, collagen and fibronectin production were assessed by specific staining, ELISA,
Sircol and immunostaining, respectively. All procedures involving care and use of laboratory animals were approved by the
Animal Ethics Committee of Fiocruz (CEUA-FIOCRUZ, Prot. 0213-4). We found that allergen provocation led to marked lung
inflammatory response - predominantly eosinophilic- and expressive signs of airway remodeling and matrix protein deposit.
These changes were accompanied by elevation in IL-13 (~150%), eotaxin-1 (~400%), IL-4 (~500%) and IL-5 (~86%) levels,
measured in lung tissue samples. We also noted that lung fibroblasts obtained from healthy mice (control lung) or actively
sensitized and challenged mice (remodeled lung) consistently evolved to spheroid clusters within 24 h of culture. Spheroids from
fibroblasts from control lungs were found to be smaller (217.95.0 m compared to 270.26.9 m (meanSEM; n=6; P
Conclusions:
In conclusion, these findings show that cultured spheroid formed by lung fibroblasts coming from allergen-challenged mice
exhibit phenotype and functional differences, related to repair and airway remodeling, reinforcing the view that changes acquired
by fibroblast experiencing remodeling events may persist in future generations of these cells, what could contribute to the
severity of asthma.
Keywords: ASTHMA, LUNG FIBROBLASTS, 3D-CULTURE
Financial Support: CAPES, FAPERJ, CNPq, PAPES and PDTIS
Resumo:17-179
CANAVALIA LECTINS: STRUCTURE VERSUS ANTINOCICEPTIVE ACTIVITY
Pinto, N. V. 1; Brito, L. F. 1; Pires, A. F. 1; Cavada, B. S. 2; Assreuy, A. M. S. 1

1
Instituto Superior de Cincias Biomdicas, ISCB-UECE
2
Departamento de Bioqumica e Biologia molecular, UFC
Objectives:
Diocleinae lectins, despite of the highly structural homology, express diverse biological activities, differing also in potency and
efficacy for the same activity. In this study it was investigated the different profiles in nociception models of lectins isolated from
Canavalia genus that posse high degree of similarity in their sequence of amino acids and three-dimensional structures.
Lectins from seeds of C. gladiata - CGL; C. maritima - ConM, C. brasiliensis - ConBr were isolated and purified by affinity
chromatography. Evaluation of antinociceptive activity was conducted in Swiss mice (25-35g), manipulated according to the
principles established by the Ethics Committee of UECE (No. 10130208-8/40). Mice received intraplantar (i.pl) injection of
formalin 20l (2.5% v / v) in the right hind paws and the time (seconds) that animal spent licking its paws was recorded at
neurogenic (F1: 0-5 min) and inflammatory (F2: 15-30 min) phases. In the Hot Plate test, the reaction latency (time displayed
before appearance of licking and shaking hind paws or jumping) of mice to thermal stimuli (55C) was recorded before and after
(30, 60, 90, 120, 150 and 180 min) lectins administration. Mechanical hyperalgesia was induced by i.p. injection of carrageenan
(Cg, 300 mg) and assessed by the use of Von Frey filaments (0.8 g). Finally, to eliminate the interference effects in areas
responsible for motor function was performed rota-rod test and the number of falls and the time that the animals remained in the
apparatus was recorded. Animals were treated with lectin (1 mg/kg) intravenously (i.v.) 30 min. before protocols. Results are
presented as mean SEM analyzed by ANOVA followed by Bonferroni test, considering p

Conclusions:
Canavalias lectins, although present structural similarity, inhibited the nociception inflammatory pain models by peripheral
mechanisms apparently by different pathways. These differences can be associated to pH-dependent oligomerization and to
substitutions of key aminoacids involved in the structure of the carbohydrate-binding sites and to quaternary structures.
Keywords: Antinociceptive activity, Canavalias, Lectin, Structure
Financial Support: CNPq, CAPES, Marques-Domingos, G.F.O.
Resumo:17-180
ANTINOCICEPTIVE EFFECT OF TEPHROSIA TOXICARIA PERS IN TEMPOROMANDIBULAR JOINT
ARTHRITIS INDUCED BY ZYMOSAN: INVOLVEMENT OF NITRIC OXIDE, ATP-SENSITIVE POTASSIUM
CHANNELS, HISTAMINE AND HEMEOXYGENASE-1.
Val, D. R. ; Filgueira, A. A. ; Gondim, C. B. ; Rios, L. C. ; Arriaga, N. C. ; Chaves, H. V. ; Silva, A. A. R. ;

Filho, G. C. ; Oliveira, J. M. ; Bezerra, M. M.
Objectives:
Temporomandibular joint (TMJ) disorders are a group of important clinical entities that result in TMJ inflammatory pain. In the
context of inflammatory responses nitric oxide, ATP-sensitive potassium channels, histamine and hemeoxygenase-1 are
important regulators. Plants of the genus Tephrosia are distributed in tropical and subtropical regions and they are used by
community in allergic and inflammatory conditions, such as asthma and rheumatism. The present study investigated the
antinociceptive effect of Tephrosia toxicaria (Tt) pers in TMJ arthritis induced by zymosan and also verified the possible role of
nitric oxide, ATP-sensitive potassium channels, histamine and hemeoxygenase-1 in this action.
42 Male Wistar rats (160-220 g) were pretreated with Tephrosia toxicaria (0,2, 2,0 or 20 mg/kg; per os) 60 min before the
injection (intra-articular-i.a) of 40 L zymosan (2 mg) into the left TMJ. Zymosan group (Zy) received saline (s.c) 60 min before
TMJ arthritis. Sham group (SH) received saline into left TMJ (i.a). Indometacin (5 mg/kg) was used as a positive control group.
Von Frey test was used to assess mechanical hipernociception (4th hour) and the animals were sacrificed 6h after Zy. In other
series of experiments, before Tt (20 mg/kg) injection, animals were pretreated with L-NAME (30 mg/kg; i.p.), a non-specific
inhibitor of the activity of nitric oxide synthase isoforms; glibenclamide (10mg/kg, i.p.), an ATP-sensitive potassium channel
antagonist; meclizine (40 mg/kg; s.c.), an inhibitor of histamine H1 receptors; or ZnPP-IX (3mg/kg, s.c.), a specific HO-1
inhibitor, followed by Zy injection (i.a). Histopathological analysis of TMJ were performed to evaluate cell influx. Results are
expressed as mean S.E.M. Histological data are expressed as medians. P
Conclusions:
These results suggest that the impairment of hypersensitivity elicited by Tt depends on the integrity of both NO and HO-1
pathway, as well as, ATP-sensitive potassium channels, and H-1 receptors.
Keywords: Antinociceptive , Arthritis, ATM, Tephrosia toxicaria, Zymosan
Financial Support: Funcap and CNPq
Resumo:17-181
CARVACROL ATTENUATES MECHANICAL HYPERNOCICEPTION IN MICE
Guimares, A. G. ; Xavier, A. M. ; Santana, M. T. ; Cavalcanti, S. C. H. ; Bonjardim, L. R. ; Camargo, E.

A. ; Antoniolli, A. R. ; Santos, M. R. V. ; Quintans-jnior, L. J.
Fisiologia/Universidade Federal de Sergipe, UFS
Objectives:
Carvacrol (5-isopropyl-2-methylphenol) is a phenolic monoterpene present in the essential oil of the family Lamiaceae, which
includes the genera Origanum and Thymus. The purpose of the present study was to evaluate the anti-hypernociceptive activity of
carvacrol (CARV) in mice.
The anti-hypernociceptive activity of CARV (25, 50 and 100 mg/Kg; i.p.) was tested in male Swiss mice (n=8/ per group)
through models of mechanical hypernociception induced by carrageenan (CG; 300 g/paw) and the involvement of important
mediators of its signaling cascade, as TNF- (100 pg/paw), PGE2 (100 ng/paw) and dopamine (30 g/paw). Mechanical
hypernociception was evaluated using digital Von Frey apparatus (Insight, Brazil), before and at 0.5, 1, 2 and 3 h after the
administration of hypernociceptive agents. The experimental protocols were approved for the UFS Ethic Committee (CEPA:
43/08). Data were evaluated by ANOVA followed Tukeys test (p
Conclusions:
Our results show that CARV reduces inflammatory hypernociception and supporting the idea of this monoterpene acts inhibiting
the sensitization of the nociceptor by anti-inflammatory pathways.
Keywords: Monoterpene, Carvacrol, Hypernociception
Financial Support: CAPES, CNPq and FAPITEC-SE
Resumo:17-182
STEM CELL FACTOR STIMULATES SMOOTH MUSCLE CELLS FROM TRACHEA TO EXPRESS TGF-ALFA.
Oliveira, Lcf ; Oliveira, Shp

Depto Cincias Bsicas/Faculdade de Odontologia de Araatuba, UNESP
Objectives:
The airways smooth muscle cells plays a central role in asthma. These structures being recognized a great potential for active
participation in airway allergic processes for the synthesis of cytokines, chemokines and adhesion molecules with capacity to
promote and perpetuate the inflammatory mechanisms in the pathology present. The Stem cell factor(SCF) is an important
component of this synthetic process, working in maintenance and survival of mast cells, inducing chemotaxis and taking crucial
role in its adhesion to the extracellular matrix. The aim of this study was to evaluate the role of SCF on the Transforming Growth
Factor-beta(TGF-beta)expression on smooth muscle cells of the trachea of mice.
We used Balb/c young male mice (six animals per group) weighing 18-20g with free access to food and water before being
subjected to testing. The animals were sacrificed for collection of the trachea that was treated, washed and perforated to obtain
the smooth muscle cells that were stimulated by SCF. The expression of TGF-beta was analysis by RT-PCR. The smooth muscle
cells were stimulated with SCF (1, 10 e 100 ng/mL) at 1, 6 e 24h. After 6h hours, SCF in the concentration of 100 ng/mL was
able to stimulate smooth muscle cells from trachea to express TGF-beta. However, 1 and 24 hours we are not able to observe any
mRNA TGF-beta expression.
Conclusions:
TGF-beta is known cytokine with a role in regulating inflammation and airway remodeling, with isoforms related to the
epithelium and smooth muscle tissues of the lung. The elucidation of the mechanisms involved in the production of TGF-beta by
smooth muscle cells of airways, as well as evaluation of possible related signaling pathways are important tools for better
understanding of the etiology of asthma and to identify potential cellular targets of more selective drugs that can be used in its
pharmacotherapy.
Keywords: Airways smooth muscle cells, SCF, TGF-beta
Financial Support: FAPESP, CAPES.
Resumo:17-183
ROLE OF FRACTALKINE EXPRESSED IN DORSAL ROOT GANGLION IN INFLAMMATORY
HYPERNOCICEPTION
Souza, G. R ; Cunha, T. M. ; Lotufo, C. M. C. ; Talbot, J. ; Bozzo, T. A. ; Cunha, F. Q. ; Ferreira, S. H.

Departamento de Farmacologia - USP-RP, FMRP-USP
Objectives:
Glial activation in the central nervous system has been implicated in the development of neuropathic and inflammatory pain.
More recently, it was found that activation of satellite cells present in dorsal root ganglion (DRG) seems also to play a role in
nociception. The chemokine, Fractalkine (CX3CL1) is expressed by primary sensory afferent, whereas its receptor (CX3CR1) is
predominantly found in the glial cells, indicating that fracktalkine could participate in the process of glial activation. This
chemokine is tethered to the extracellular surface of neurons and when released constitute a diffusible signal that contributes to
neuron-glial interaction. The aim of this study was to test whether activation of satellite cells by fractalkine is involved in the
cascade of events responsible for the genesis of the inflammatory pain.

Mechanical nociceptive threshold was evaluated with an electronic version of von-Frey test (electronic pressure-meter paw test)
in Wistar rats weighing 150-200 g. Firstly, it was observed that the decrease in mechanical nociceptive threshold
(hypernociception) observed during peripheral inflammation of rat paw by intraplantar injection of carrageenan was inhibited by
the anti-CX3CR1 administered into the dorsal root ganglion (DRG-L5). Furthermore, intraplantar injection of carrageenan
induced significant increase in GFAP mRNA expression. This increase in GFAP mRNA expression was blocked by the treatment
with a specific antibody against CX3CL1 (10g/i.gl). In agreement, direct administration of fractalkine into the DRG (L5) of rats
produced mechanical hypernociception of hind paws in a dose- and time- dependent manner. Regarding the mechanism by which
glanglionar fractalkine mediates inflammatory hypernociception, it was observed that fractalkine hypernociceptive effect was
blocked by the treatment with a specific antibody against CX3CR1 (10g/i.gl) and CX3CL1 (10g/i.gl), infliximab (50ng/i.gl.),
IL1-ra (300ng/i.gl.), indomethacin (50g/i.gl) as well as dexamethasone (20ng/i.gl). In vitro, incubation of isolated satellite cells
in culture with fractalkine induced the release of TNF-alpha and IL-1beta.
Conclusions:
Overall, these results suggest that during peripheral inflammation fractalkine is released in DRG and contributes to the genesis of
inflammatory hypernociception by a mechanism dependent on stimulation of satellite cells to produced cytokines (TNF-alpha and
IL-1beta). Finally, these cytokines might activate COX enzyme triggering the release of prostanoids which are responsible for the
nociceptor sensitization and possibly maintaining inflammatory pain.
Keywords: dorsal root ganglion, fractalkine, glial cell, hyperalgesia, satellite cell
Financial Support: FAPESP, CNPq e FAEPA
Resumo:17-184
ANTINOCICEPTIVE AND ANTI-INFLAMMATORY EVALUATION OF NEW SUBSTANCES DERIVED FROM
ISATIN
Gonalves, M. R. 1; Zardo, R. S. 1; Silva, B. V. 2; Pinto, A. C. 2; Fernandes, P. D. 1

1
Laboratrio de Farmacologia da Inflamao e do xido Ntrico, ICB - UFRJ
2
Instituto de Qumica, IQ-UFRJ
Objectives:
Isatin (1H-indole-2,3-dione) is a synthetically versatile substance employed for the synthesis of a large variety of compounds and
with several biological effects. The objective of this work was to evaluate the antinociceptive and anti-inflammatory properties of
new substances derived from isatin.
Four substances derived from isatin (named Isaox 1, 2, 3, and 4) were tested in the licking response induced by formalin (2.5%,
intraplantar), acetic acid induced abdominal writhing (2%) and in the subcutaneous air pouch (SAP) model. Male swiss mice (2025g, n=4-5) received oral administration of the isatin derivatives (0.1 to 30 mg/kg), 1h before formalin or acetic acid injection. In
SAP, animals received the substances (30 mg/kg) 1h before and 23h after carrageenan injection (1% in SAP). The use of animals
was approved by the ethical committee of animal experimentation from Centro de Cincias da Sade (UFRJ), and received the
number ICBDFBC015. Statistical analyses was performed by ANOVA and Bonferronis test (*p
Conclusions:
These results suggest that the news substances present anti-inflammatory and antinociceptive effects. Thus, pharmacological
studies are continuing in order to characterize the mechanism(s) responsible for the antinociceptive and anti-inflammatory action.
Keywords: anti-inflamatory, antinociceptive, evaluation, isatin, new derivatives
Resumo:17-185
PROTEINASE ACTIVATED RECEPTOR-4 (PAR-4) EXPRESSED IN SKIN MAST CELLS AND PRURICEPTIVE
NEURONS MEDIATES ITCH-LIKE BEHAVIOUR IN MICE.
Patricio, E. S. ; Costa, R. ; Figueiredo, C. P. ; Calixto, J. B.

Farmacologia/Universidade Federal de Santa Catarina, UFSC
Objectives:
Itch is a common symptom present in several cutaneous and systemic diseases. Recently, it has been suggested a role for PAR-4
in itch transmission (J Neurosci, 28: 4331, 2008), and PAR-4 agonists induced itch-like behaviour in mice (J Pharmacol Sci., 108:
385, 2008). The present work aims to investigate cellular and pharmacological mechanisms underlying the pruriceptive actions of
the selective PAR-4 agonist AYPGKF-NH2 (AYP) in mice.
Methods: Female adult CD-1 mice (25-30 g, n=6) received a dorsal intradermal (i.d.) injection of AYP (30-500 nmol/site), and
the frequency of scratching bouts with the hind paws toward the injected site was quantified by 30 min (Ethics Committee
protocol number: PP00497). Results: I.d. administration of AYP (30-500 nmol/site) elicited a marked and dose-related scratching
behaviour. The effective dose ranged from 200 to 500 nmol/site [p<0.05]).
Conclusions:
These findings provide new evidence that PAR-4 activation on mast cells and pruriceptive neurons induces scratching behaviour
in mice. In spite of the mast cell involvement, the itching elicited by PAR-4 activation seems to be independent of histamine,
serotonin or mast cell protease release.
Keywords: gastrin-releasing peptide, mast cells, PAR-4, scratching behaviour
Financial Support: CAPES/CNPq/FAPESC.
Resumo:17-186
COMPARATIVE STUDY OF ENZYMATIC, IMMUNOCHEMICAL AND BIOLOGICAL PROPERTIES OF
BOTHRIECHIS SCHLEGELII SNAKE VENOM FROM COLOMBIA AND COSTA RICA
Prezotto-neto, J. P. ; Kimura, L. F. ; Gutirrez, J. M. ; Otero, R. ; Santoro, M. L. ; Barbaro, K. C.

Instituto Butantan, IBu
Objectives:
Bothriechis schlegelii is a relatively small snake found in Mexico, Central America and northwest of South America. Bites in
humans are characterized by pain, edema and ecchymosis at the site of bite, and slight defibrinating effect. The aim of this work
was to carry out a comparative study of enzymatic and immunochemical properties and toxic activities of B. schlegelii snake
venom from Colombia (BsCo) and Costa Rica (BsCR). Furthermore, this work also investigated the efficacy of polyspecific antiBothrops serum (ABS), produced by Butantan Institute, to recognize and neutralize some toxic activities of BsCo and BsCR
venoms.
Silver stained SDS-PAGE (12 %) was used to compare the protein profile of BsCo and BsCR venoms (5 g). Many components
with similar molecular masses between 150 22 kDa were observed in both venoms, but some protein bands of 62, 33, 25 and 22
kDa were observed exclusively in BsCo venom. Zymography was employed to detect proteolytic and hyaluronidase activities of
venoms, using fibrinogen (0.5 mg/mL), gelatin (2 mg/mL), casein (2 mg/mL) or hyaluronic acid (170 g/mL). Fibrinogenolytic
activity was observed using 40 g of BsCo venom (bands of approximately 46, 42 and 30 kDa) and BsCR venom (band around
43 kDa). Different profiles of gelatinolytic activity were also observed using 20 g of BsCR (bands in 43 and 40 kDa region) and
BsCo (bands between 54 - 25 kDa). BsCo and BsCR venoms exhibited caseinolytic spots between 50 23 kDa. Hyaluronidases
band of 60 kDa was observed only in BsCR venom (40 g). Intense cross-reactivity between BsCo and BsCR was detected by
ELISA using ABS. Many different components located around 150 and 15 kDa in BsCo and BsCR venoms were recognized
using ABS by Western blotting. BsCo and BsCR venoms (0.5, 1, 2, 4 and 8 g) were injected in mice footpad (Swiss, n= 6) to
evaluate edema evolution in 0.5, 1, 2, 4, 24, 48 and 72 h. The highest edematogenic activity of venoms was noticed at 30 min.
However, BsCR venom was more edematogenic when compared to BsCo venom, since the edema induced by 8 g dose persisted
up to 48 h. The edema induced by BsCo and BsCR venoms was partiality neutralized after pre-incubation with ABS in the first 4
h. The ABS also neutralized around 87 % of the edema induced by BsCR venom (8 g) in 48 h. Both BsCo and BSCR venoms
induced hemorrhage 2 h after i.d. injection in mice dorsum (n= 6). However, BsCR venom was almost 4 times more hemorrhagic
when compared with BsCo venom. ABS (80, 40 and 20 L) totally neutralized the hemorrhagic activity induced by BsCo and
BsCR venoms.
Conclusions:
Geographic distribution can influence the composition and activity of B. schlegelii venoms. High antigenic cross-reactivity was
observed in both venoms (BsCo, BSCR) using ABS. Moreover ABS was effective to neutralize some toxic activities from BsCo
and BsCR venoms.
Keywords: Bothriechis schlegelii, veonom, antivenom, edema
Financial Support: FAPESP, CAPES
Resumo:17-187
STUDY OF PARAOXONASE 1 AND 2 IN PATIENTS WITH SICKLE CELL DISEASE.
Macedo, C. G. D. 1; Maselli, L. M. F. 1,3; Gualandro, S. F. M. 2; Fonseca, G. H. H. 2; Chamone, D. D. A. F.
; Bydlowski, S. P. 1
1
Laboratrio de Gentica Molecular e Hematologia/Fac Medicina, LIM-31 FMUSP
2
Servio de Hematologia do Hospital das Clnicas da FMUSP, HCFMUSP
3
Fundao Pr-Sangue, Hemocentro So Paulo
Objectives:
The prevalence of sickle cell trait in Brazil is estimated as ranging from 2-8%. Anemia due to this trait presents extremely
heterogeneous clinic. Among other events, there is inflammation and changes related to the redox phenomena. PON1 is an
enzyme present in serum, strongly associated with apoA-I/HDL. Its primary physiological role is protect LDL against oxidative
modifications.The association between inflammatory diseases and the decreases of apo A1 and apo A2, whose are related to the
concentration and activity of PON1, are known. However, a significant reduction of apo-A1 during sickling crisis was already
reported. Polymorphisms in candidate genes that affect vascular and inflammatory components involved in the mechanisms of
disease may have prognostic utility. Among them, the PON family of enzymes has been widely studied concerning to protection
against oxidation and its behavior in the inflammatory processes. The present study aims to evaluate, in patients with sickle cell
disease, arylesterase activity of PON1, as good as PON1 and PON2 polymorphisms.
Genomic DNA was obtained from 5mL of whole blood from 16 patients with sickle cell disease and 20 from healthy people. The
polymorphisms were assayed by RFLP-PCR using Hinf I. The restriction product was analyzed in 2% agarose gel stained with
ethidium bromide. Arylesterase activity of PON1 was performed by specthrophotometry using phenylacetate in kinetic enzyme
method. The lipid profile (total cholesterol, HDL, LDL and VLDL cholesterol, triglycerides and apo A1) was determined as
described worldwide. Results showed an allelic frequency of 62,5% and 37,5% (controls), 43,75% and 56,25% (patients) for the
alleles PON1-192Q and PON1-192R; 60% and 40% (controls), 84,38% and 15,62% (patients) for the alleles PON1-55L and
PON1-55M; and 85% and 15% (controls), 84,38% and 15,62% (patients) for PON2-148A and PON2-148G. Arylesterase activity
of PON1 in patients showed mean of 70(29) U/mL and the control showed mean 93,9(26,52). Means observed for total
cholesterol were 128,7(26,39)mg/dL; 34,35(11,22)mg/dL for HDL-cholesterol; 69,7(22,1)mg/dL for LDL;
24,64(8,53)mg/dL for VLDL; 123,23(43,16)mg/dL for triglycerides; and 107,44(21,88)mg/dL for apo A1. This study was
approved by the ethics committee CAPPesq (0285/10).
Conclusions:
The distribution of alleles PON1-192Q and PON1-192R and PON1-55L and PON1-55M differed between groups, while PON2148A and PON2-148G showed similar frequencies. The values of apo A1 and HDL, in patients, were reduced, a finding that
could contribute to increased inflammatory activity in these patients.Hemolytic stress could be associated with a significant
reduction in plasma lipids and lipoproteins. It appears that patients with sickle cell anemia are at a lower risk for coronary artery
disease.
Keywords: Inflammation, PON1, PON2, Sickle cell
Resumo:17-188
ANALYSIS OF THE ANTINOCICEPTIVE ACTIVITY OF &BETA-IONONE
Freitas, L. B. N. 1; Luz, P. B. 1; Osrio, C. B. D. H. 1; Olinda, T. M. D. 1; Sousa, T. D. F. G. D. 1; Carmo, L.

D. D. 1; Alencar, N. M. N. D. 1; Sousa, D. P. 2

Departamento de Fisiologia, UFS
Objectives:
The &beta-ionone (4 - [2,6,6-cyclohexene-1-trimetill]-3-butene-2-one) is a sesquiterpene (degraded terpenoid - C13) present in
the molecular structure of retinol, &beta-carotene and acid retinoic, formed from the mevalonate pathway in different types of
plants. The research on the biological activities of &beta-ionone are still incipient and limited results with this compound are
found in the literature. The objective of this study was to investigate the effect of &beta-ionone in classic models of nociception.
Animal handling and experimental protocols were approved by the Ethical Committee for Animal Research/UFC under number
0286. To this end, we used female Swiss mice (n = 10, 23 2g). The animals were treated 30 minutes before each experiment
with &beta-ionone (12.5, 50 and 200mg/kg i.p.), Tween 1% (vehicle 10ml/kg i.p.), morphine 5mg/kg s.c. or diazepam 2mg/kg
s.c. Writhing Abdominal were induced by acetic acid 0.85% (10ml/kg i.p.) and after 10 min, the number of writhing were
counted for 20 min. In the formalin test, the time at which the animal is still licking the injected paw with formalin (1,2%, 20 mL
/ paw, s.c.) was recorded in the first 5 min (1st phase) and 20-30 minutes (2nd phase). In the hot plate test, reaction time of
animals was monitored at 55 C at 0, 30, 60, 90, 120 and 150 min. Rota Rod test was performed in order to study a possible
activity of &beta-ionone on the locomotor system, mice were placed in the swivel bar (4 rpm) and the residence time was clocked
in 2 minutes. The &beta-ionone (50 e 200mg/kg) significantly reduced the writhing induced by acetic acid, compared with
vehicle (8.33 2.46, 0.75 0.41 and 39.6 3, 8, respectively). In the formalin test, the &beta-ionone (200mg/kg) significantly
reduced response compared with the vehicle, in the 1st phase (3.14s 1.03, 63s 6.58, respectively), whereas in 2nd phase of the
test, all doses of &beta-ionone (12.5, 50 and 200mg/kg) were able to reduce the time to lick when compared to vehicle (45.8s
7.66, 38.8s 8.75, 0s and 121s 14.5 respectively). In the hot plate test, the &beta- ionone (200 mg/kg) significantly increased
the reaction time of animals at 30 and 60 min. (27.1s 3.8, 22.4s 3.1, respectively) compared with vehicle (11.7s 1.1, 12.1s
1.8, respectively). In Rota Rod, the &beta-ionone did not alter the time spent in the swivel bar. Statistical analysis was performed
by ANOVA followed by Student Neuman-Keuls.
Conclusions:
The results show the &beta-ionone has antinociceptive activity, by peripheral and central mechanism of action to be clarified.
Further researches are being developed to explain the exact mechanism of these effects.
Keywords: &beta-ionone, nociception, sesquiterpene
Resumo:17-189
PARADOXICAL EFFECTS OF BRAIN DEATH AND ASSOCIATED TRAUMA ON RAT MESENTERIC
MICROCIRCULATION: AN INTRAVITAL MICROSCOPIC STUDY.
Simas, R. ; Sannomiya, P. ; Cruz, J. W. M. C. ; Correia, C. J. ; Zanoni, F. L. ; Menegat, L. ; Moreira, L. F.

P.
Depto Cardiopneumologia FMUSP, InCor HC-FMUSP
Objectives:
Aim:To evaluate the role of brain death (BD) compared with BD-associated trauma on the development of the inflammatory
response at the mesenteric microcirculation and its systemic effects.
Methods and Results: Male Wistar rats (weighting 250-300 g) were anesthetized with isoflurane 5-2 %, intubated and
mechanically ventilated with a tidal volume of 10 mL/kg. Carotid artery was accessed to measure mean arterial pressure (MAP),
heart rate (HR) and blood sampling. Jugular vein was used to administration of saline 2 mL/h. To induce BD, a balloon catheter
was placed intracranially and rapidly inflated with 500 L of water. BD was confirmed by sharp rise of MAP, maximal pupil
dilatation, absence of reflex and apnea. Sham operated animals were trepanned only. Animals (7 per group) were evaluated 30
and 180 minutes thereafter. The mesenteric microcirculation was analyzed by intravital microscopy in situ. Mesentery was
exposed and continually perfused by Krebs-Henseleit solution (pH 7.2-7.4), and three fields were obtained to quantify percentage
of perfusion and number of roller, adhered and migrated leukocytes. Expression of adhesion molecules, P-Selectin and ICAM-1,
was performed by immunohistochemistry of mesentery. Serum concentration of TNF-, IL-1, IL-6, IL-10, CINC-1 and CINC-2
was determined by ELISA. Number of total and differential white blood cells was determined. The methodology was approved
by the ethics committee (CAPPesq 0226/09). After induction of BD, it was observed an immediate increase in MAP values,
followed by hypotension. MAP values did not change over time in Sham rats. There were no differences in HR values among
groups. Proportion of perfused small vessels (30 m diameter) was reduced in BD-rats compared to Sham rats either at 30
minutes (BD 30.20.06 % vs Sham 77.90.07 %, p
Conclusions:
Conclusion: BD-associated trauma is responsible for most of the inflammatory events observed. Otherwise perfusion of
mesenteric microvessels was promptly interrupted by BD itself. This was accompanied by a pronounced leucopoenia, and
increased ICAM-1 expression and migrated leukocytes.
Keywords: Brain Death, Cytokines, Microcirculation
Financial Support: Financial support: FAPESP.
Resumo:17-190
CENTRAL INTERACTIONS BETWEEN SUBSTANCE P AND PROSTAGLANDINS DURING FEVER.
Brito, H. O. ; Reis, R. C. ; Barbosa, F. L. ; Fraga, D. ; Zampronio, A. R.

Dept Pharmacology, UFPR
Objectives:
We showed previously that the neuropeptide substance P (SP), acting on NK1 receptors is involved in the febrile response
induced by lipopolysaccharide, tumor necrosis factor- (TNF-) and interleukin(IL)-6 but does not participate in the fever
induced by IL-1 and macrophage-inflammatory protein-1. Since TNF- and IL-6 induce a prostaglandin-dependent febrile
responses we decided to investigate the involvement of prostaglandins (PG) in the febrile response induced by SP.
Male Wistar rats (200 g) were implanted with a guide cannula in lateral ventricle and with data loggers (for measurement of body
temperature in the peritoneal cavity) one week before the experiments under the same anesthesia. Body temperature (C) was
registered every 15 min. The room temperature was kept at 28C. All the methods were previously approved by Institutions
Ethics Committee on Animal Use under protocol # 384. Central administration of SP (250, 500 e 750 ng/2l) induced a dosedependent increase on body temperature in captopril-treated rats (5 g/2l, 30 min before). The febrile response induced by
500ng SP was completely blocked (100%) by the non-peptide NK1 receptor antagonist SR140333 (3.0 g/2l, icv). To evaluate
the involvement of PG in this response rats were treated with PG-synthesis inhibitor indomethacin (IND, 2 mg/kg, i.p.) or the
same volume of vehicle (Tris buffer, pH 8.2). After 30 min, animals received captopril (5 g/2l) followed by an i.c.v. injection
of SP (500 ng) or vehicle (2 l) 30 min later. As a positive control, another group of animals were treated with the same dose of
IND or vehicle and after 30 min they received IL-1 (3.1ng, i.c.v.). SP and IL-1 produced an increase in body temperature of
about 1 and 1.5C, respectively, which peaked at 2h. IND reduced the febrile response induced by both SP (53%) and IL1(86%). In a final set of experiments, animals were treated with SR140333 (3.0 g/2l, icv) and after 30 min they received an
i.c.v. injection of PGE2 (250 ng/2l) or vehicle (2 l). PGE2 induced an increase of 1.5C in the body temperature in the first 30
min after injection. Surprisingly, SR140333 abolished (100%) the febrile response induced by PGE2.
Conclusions:
Our results show that SP,administered in the central nervous system, induces fever in rats when its metabolism is blocked by
captopril. The febrile response induced by SP seems to be dependent on PG synthesis, similarly to the response induced by TNFand IL-1. However, the release of SP may also be important after PG synthesis, although further studies are necessary for the
full understanding of those interactions.
Keywords: febrile response, prostaglandin, substance P, NK1 receptor, SR140333
Financial Support: CNPq, Fundao Araucria, CAPES and Sanofi-Aventis.
Resumo:17-191
STUDY OF MESENTERIC MICROCIRCULATORY ALTERATIONS ON BRAIN DEAD RATS.
Kase, M. ; Sannomiya, P. ; Simas, R. ; Cruz, J. W. M. C. ; Moreira, L. F. P.

Depto. Cardiopneumologia, FMUSP
Objectives:
Aim: To evaluate the role of brain death (BD) on mesenteric microcirculatory perfusion, by real-time observation.
Methods and Results: Male Wistar rats (250-350g, 2 months of age) were anesthetized with isoflurane (2-5%) and intubated. A
balloon catheter was placed into intracranial cavity and inflated to induce BD. Mesenteric microcirculation was analyzed by
intravital microscopy (5 BD-induced rats) 30 minutes before, during BD, and 30 minutes thereafter. BD was confirmed by sharp
rise of mean arterial pressure, maximal pupil dilatation, absence of reflex and apnea. The mesenteric microcirculation was
analyzed by intravital microscopy in situ. Mesentery was exposed and continually perfused by Krebs-Henseleit solution (pH 7.27.4). Five fields (1 mm2/field) of mesenteric microcirculation were randomly selected for analysis. Vessels (30 m and >30
m diameter) were evaluated in each field, and classified as perfused or non-perfused microvessels (0226/09 InCor-HC-FMUSP).
Before BD, perfusion was presented in 856 % of total vessels (30 m). After induction of BD, there was a reduction to
425 % of total vessels (30 m; p<0.001).
Conclusions:
Conclusion: The perfusion of mesenteric microvessels was promptly interrupted by BD itself, and was restricted to 30 m
diameter vessels compromising, therefore, the viability of the mesentery.
Keywords: Brain Death, Intravital Microscopy, Microcirculation
Financial Support: Financial support: FAPESP.
Resumo:17-192
EFFECTS OF HYDROGEN SULFIDE (H2S) ON LUNG ALLERGIC RESPONSE IN MICE
Benetti, L. R. 2; Campos, D. 2; Almeida, M. F. 2; Guedes, C. E. V. 2; Barillas, S. G. 1; Antunes, E. 1;

Ferreira, H. H. D. A. 2
1
FARMACOLOGIA, UNICAMP
2
UNIVERSIDADE SO FRANCISCO, USF
Objectives:
Hydrogen sulfide (H2S), like nitric oxide (NO), is a newly found gasotransmitter recently demonstrated to play an important role
in inflammatory diseases. H2S may react with reactive oxygen and/or nitrogen species limiting their toxic effects. It has also been
suggested that H2S may scavenge the excess of NO produced in the inflammatory state. Thus, endogenous H2S might be an
antioxidant in our organism, but its contribution to the pathophysiology of lung allergic diseases, such as asthma, is unclear. The
aim of this study was to investigate the effect of H2S on antioxidant enzymes and on nitric oxide inducible isoform (iNOS)
expression. We also compare the consequence of H2S exposition with the anti-inflammatory effect of iNOS inhibition on the
influx of eosinophils to the lungs of allergic mice.
Methods: BALB/c mice, previously sensitized with ovalbumin (OVA; 100 mg), were treated intraperitoneally (i.p.) with a H2S
donor, NaHS (14 mol/kg), or iNOS inhibitor, 1400W (1.0 mg/kg), 30 minutes and 2 hours before OVA challenge (10 g),
respectively. Control (non-treated) mice received only saline i.p. Twenty-four, 48, 96, 120 and 144 hours after challenge, the
mice were sacrificed, bronchoalveolar lavage (BAL) was collected and the lungs were removed and homogenized. The total cell
numbers in BAL were determined and differential leukocyte counts were carried out using cytospin preparation of the cell
suspension, stained with Diff-Quick. The expression of enzymes MnSOD, Cu/Zn SOD and iNOS in the lung homogenates were
analyzed by Western blotting. All experiments were approved by the EAC/USF Protocol (002.11.08). Results: The analyses of
leukocyte population present in the BAL of the control group showed that eosinophil infiltration had started by 24 h and that a
marked increase beginning at 48 h had peaked at 96 h. This effect was not sustained since the eosinophil number in the BAL at
120h was similar to that seen at 48 h, but another peak of this cell influx to lungs was observed at 144h after OVA-challenge.
Treatment of mice with 1400W or NaHS resulted in an approximately 50% decrease in eosinophil infiltration at 48h and 144 h.
At other times after challenge, the eosinophil numbers in the BAL were not significantly different from the control. The analysis
of lung enzymes by Western blotting showed that the MnSOD expression was not modified by OVA challenge neither by NaHS
treatment. In addition, the expression of Cu/Zn SOD was significantly (p
Conclusions:
Our results suggest that the beneficial action of H2S on the mice allergic response by inhibiting the migration of eosinophils to
the lungs is similar to those resulted from iNOS inhibition. This effect may not depend on the antioxidant enzyme, MnSOD or
Cu/Zn SOD. The involvement of iNOS in this process needs to be better clarified by examination of iNOS activity in the
presence of NaHS
Keywords: ASTHMA, HYDROGEN SULFIDE, NITRIC OXIDE
Resumo:17-193
TREATMENT WITH H2S DONOR AND INOS INHIBITOR ATTENUATES THE INFLAMMATORY RESPONSE IN
THE LUNG OF ALLERGIC MICE
Guedes, C. E. V. ; Rocha, T. ; Almeida, M. F. D. ; Ferreira, H. H. D. A.

UNIVERSIDADE SO FRANCISCO, USF
Objectives:
Pathological features of bronchial asthma are characterized by an increase in inflammatory cells, mainly eosinophils, associated
with plasma exudation, oedema, smooth muscle hypertrophy, mucus plugging and epithelial damage and increases in expression
of nitric oxide inducible isoform (iNOS). Unlike other chronic obstructive lung diseases, asthma can be controlled and reversible.
The objective of this study was to analyze the effect of sodium hydrosulfide (NaHS), a H2S donor, and N-3-aminomethyl-benzylacetamidine-dihychloride (1400W), an iNOS inhibitor, on the inflammatory response in the lungs of allergic mice.
Methods: All experiments were approved by the animal ethics committee of USF (protocol 0021108). BALB/c mice were
sensitized with subcutaneous injection of ovalbumin (OVA) bound to aluminum hydroxide, followed by a booster injection 7
days after. At days 14 to 17, the mice received an OVA intranasal challenge, twice a day. The non-challenged group (NC)
received only saline intranasally. The challenged mice were subdivided into: 1) group treated with 1400W (1mg/Kg), i.p., 2 h
before each OVA challenge; 2) group treated with NaHS (14mol/Kg), i.p., 30 min before each OVA challenge and 3) control
group (non-treated) that received only sterile saline i.p. At 24h, 48h and 96h after the first OVA-challenge, mice were sacrificed
and the left lungs were fixed in 10% formalin overnight and embedded in paraffin. Tissue slides (5m) were stained with HE to
assess the inflammatory cell infiltrate, which was defined as the average of the peribronchial and perivascular inflammation
scores, determined on a subjective scale of 0 (no detectable inflammation) to 3 (most bronchi or vessels surrounded by a thick
layer of inflammatory cells). To quantify the mucin production, the slides were stained with periodic acid Schiff (PAS) and the
goblet cells were counted. The percentage of PAS-positive cells was determined in bronchus transversal sections. Mucus
plugging was graded as 0 (1% positive cells) to 5 (1% negative cells). Results: Inflammatory cell infiltrate was not observed in
the NC group. OVA-challenged groups showed airway infiltration of neutrophils, eosinophils, mast cells and macrophages.
However the degree of histopathologic inflammation was qualitatively decreased in NaHS and 1400W-treated animals, as
compared to control, especially at 48h after OVA-challenge. At 96h, the effect of 1400W was reduced although the effect of
NaHS remained active. Percentages of goblet cells were increased in control mice, with the presence of mucus pellet into
bronchus lumen. An expressive reduction in the percentage of goblet cells was observed in the airways from the NaHS or
1400W-treated groups and no mucus pellet was observed. No increase in the number of goblet cell or mucus pellet was present in
the airways of NC mice.
Conclusions:
NaHS and 1400W both efficiently decrease the inflammatory infiltrate. As mucin production is an indicator of airway
obstruction, the reduction in mucus plugging observed in the NaHS and 1400W groups suggest the efficiency of both treatments
at attenuating asthmatic symptoms.
Keywords: H2S, iNOS, LUNGS, INFLAMMATION, MUCUS
Financial Support: CAPES, FAPESP
Resumo:17-194
DUAL EFFECT OF H1 AND H3 HISTAMINE RECEPTORS IN THE TRAFFIC OF THE NOCICEPTIVE PATHWAY
Stein, T. ; Souza-silva, E. ; Tonussi, C. R.

Farmacologia/Universidade Federal de Santa Catarina, UFSC
Objectives:
Spinal cord is known to be a center that controls ortodromic (nociception) as well as antidromic traffic (vasodilation) in the
nociceptive pathway. Histamine receptors are known to participate in the spinal cord nociceptive transmission, thus it is
conceivable that histamine may also play a role in the spinal cord control on the peripheral vasodilation. Aim: Evaluate the role of
spinal cord histamine in the articular incapacitation, edema and cell migration induced by carrageenan injection into rat kneejoints.
Male Wistar rats (300 - 400 g), received intrathecal injection of histamine (0.002; 0.2; 2; 20 nmol), the H1 agonist 2pyridylethylamine dihydrochloride (2-PEA; 0.01; 0.1; 1; 10 nmol), the H3 agonist immepip (4; 8; 16; 32 nmol), and the H3
inverse agonist thioperamide (0.0004; 0.04; 4 nmol) were given 20 min before knee-joint carrageenan injection (50 g). Articular
incapacitation was measured by counting the paw elevation time (PET; s) during 1-min periods of forced walk hourly, until 6
hours after carrageenan injection. Edema was accessed by the metering of the articular diameter (mm). After 6 h, synovial fluid
was collected for the evaluation of leukocyte infiltration (CEUA/PP00368/2009). Histamine decreased the incapacitation (20
nmol; p
Conclusions:
Histamine may act in the spinal cord by H1 and H3 receptors to cause hyponociception, but this action may also increase
peripheral inflammation.
Keywords: edema, histamine, intrathecal, pain, nociception
Financial Support: CNPq; CAPES; FAPESC
Resumo:17-195
EFFECT OF TALIDOMIDE ANALOGUE, LASSBIO-468, ON EXPERIMENTAL SILICOSIS IN MICE.
Ramos, T. J. F. 1; Trentin, P. G. 1; Ferreira, T. P. T. 1; Arantes, A. C. S. 1; Pires, A. L. A. 1; Lima, L. M. 2;
Barreiro, E. J. 2; Martins, M. A. 1; Silva, P. M. R. 1

1
Laboratrio de Inflamao / Instituto Oswaldo Cruz, IOC/Fiocruz
2
Laboratrio de Avaliao e Sntese de Substncias Bioativas, LASSBio/UFRJ
Objectives:
Silicosis, one of the oldest occupational diseases in the world, is consequence of a long-term exposure to inhaled dust containing
silica in its free and crystalline form. Lung interstitial inflammation and fibrosis are the main features of the disease, involving a
wide range of chemical mediators such as TNF-&alpha. This is a pleiotropic molecule which exerts its effects on many cell types.
LASSBio-468 is a thalidomide analogue which modulates TNF-&alpha production and inhibits endotoxic shock and arthritis in
animal models. In this study we investigated the potential effect of LASSBio-468 on the experimental model of silicosis in mice.
Anesthetized male Swiss-Webster mice (18-20g) received intranasal (i.n.) instillation of silica (10 mg/50 L) and vehicle (saline).
Treatment consisted of oral administration of the LASSBio-468 (50 and 100 mg/kg) during 7 consecutive days, from day 21 to 28
post-silica. The analyses included lung function (resistance and elastance) and airways hyperreactivity to aerosolized metacholine
(3 -27 mg/mL) were measured by whole body invasive plestimography (Finepoint, Buxco System). Morphological and
morphometrical analyses included classical dyes such as Hematoxylin-Eosin and Picrus-Sirius. Collagen content and
cytokine/chemokine generation were quantified by Sircol technique and ELISA, respectively. All experimental procedures were
performed in accordance with the guidelines of the Committee on Use of Laboratory Animals of the Oswaldo Cruz Foundation
(L-034/09. We showed that silicotic mice exhibited increased basal levels of lung resistance and elastance as well as airways
hyperreactivity to methacholine aerosolization. Tissue inflammatory response, extensive collagen deposition, granuloma
formation and chemokine (KC and MCP-1)/cytokine (TGF-&beta and IFN-&gamma) generation were also detected in the
silicotic lungs. Administration of LASSBio-468 into silicotic mice reduced the lung function compromise and airways
hyperreactivity, tissue collagen deposition (from 198,3 3,5 to 153,5 7,7 g/right lung in non-treated and treated silicotic mice,
respectively) (mean SEM; n=7) and granulomatous area (from 37,8 2,4% to 18,0 2,6% in non-treated and treated silicotic
mice, respectively; n=6). The generation of cytokines and chemokines was also suppressed by the drug.
Conclusions:
Altogether our findings show that the treatment of silicotic mice with LASSBio-468 reduced curatively the lung function
compromise and granulomatous response, indicating this compound seems to constitute a promising tool for the treatment of
chronic fibrotic pulmonary diseases such as silicosis.
Keywords: Silicosis, Inflammation, Lung, Fibrosis, Therapy
Financial Support: FIOCRUZ, CNPq, FAPERJ and CAPES
Resumo:17-196
EFFECT OF ANETHOLE AND ESTRAGOLE IN MICE PAW EDEMA INDUCED BY BRADYKININ, SUBSTANCE P
AND SODIUM NITROPRUSSIDE
Sousa, P. L. 1,2; Ponte, E. L. 1,2,3; Rocha, P. A. M. V. 1,2; Coelho-de-sousa, A. N. 1,2; Leal-cardoso, J. H. 1,2;
Assreuy, A. M. S. 1,2
1
Instituto Superior of Biomedical Sciences , ISCB
2
State University of Cear, UECE
3
Faculty Christus, FC
Objectives:
Anethole and estragole are position isomers belonging to the terpenoid group and natural constituents of essential oil of aromatic
plants. Preliminary results from our laboratory demonstrated that the essential oil of Croton zehntneri and its major constituents,
anethole and estragole, present anti-inflammatory activity inhibiting the paw edema induced by carrageenan (Cg), histamine,
compound 48/80 and serotonin. The aim of this research was to evaluate the potential anti-inflammatory effect of anethole and
estragole in the model of paw edema induced by bradykinin, substance P and sodium nitroprusside.
Swiss mice (25-30g) were manipulated in accordance with the principles of our Ethic Comitee in Research (UECE No.10 13
0208-8/40). The paw edema was induced by subcutaneous (s.c.) intraplantar injection of 50 microL bradykinin (3 nmol),
substance P (20 nmol) and sodium nitroprusside (10 micromol). Animals were treated by gavage with anethole or estragole (10
mg/kg) 60 min before inflammatory stimuli. Positive controls received stimuli and negative controls, sterile saline (NaCl 0.9%;
100 microL/10g). Edema was evaluated by hydroplethysmometry measuring the volume displaced (microL) by paws before (time
zero) and from 30 min to 240 min after stimuli and calculated by the difference between measures. Data were expressed in
microL or area under curve (AUC - arbitrary units). Results: The edema induced by bradykinin was inhibited by anethole at 30
and 60 min, about 55% (BK: 1256 179; anethole: 563 93 AUC). However, estragole inhibited the edematogenic effect only at
30 min about 56% (BK: 4830 703; estragole: 2138 518 AUC). The edema induced by substance P was inhibited by anethole
and estragole, respectively at 30, 60 and 120 min. about 38% by anethole (substance P: 7286 370; anethole: 4543 850 AUC)
and about 63% by estragole (substance P: 6666 373; estragole: 3949 225 AUC). The edema induced by a nitric oxide donor
sodium nitroprusside was reduced by estragole at 120 min. about 30% (sodium nitroprusside: 19838 1460; estragole: 13463
1412 AUC), but not by anethole.
Conclusions:
Anethole and estragole present antiedematogenic activity following a common mechanism that involves inhibition of
inflammatory peptide mediators. However, the mechanism of the inhibitory effect of estragole includes nitric oxide, suggesting
that the structural difference between these molecules influence the response.
Keywords: ANETHOLE, ESTRAGOLE, PAW EDEMA
Financial Support: CAPES, FUNCAP; CNPq and College Christus. The laboratory technique Gabriela F. O
Resumo:17-197
EVALUATION OF THE ANALGESIC EFFECT OF VENLAFAXINE, A SEROTININ AND NORADRENALINE
REUPTAKE INHIBITOR.
Magalhes, R. A. 1; Felix, J. A. 1; Albuquerque, J. C. 1; Bastos, M. V. R. 2; Viana, G. S. D. B. 2; Fonteles,

M. M. D. F. 1; Flix, F. H. C. 3; Fontenele, J. B. 1
1
Farmcia / Faculdade de Farmcia, Odontologia e Enfermagem, UFC
2
3
Hospital Infantil Albert Sabin, HAIS
Objectives:
The aim of this study was to test the analgesic effects of the antidepressant venlafaxine in several experimental models of
nociception.
Acetic acid-induced writhing: Groups of six to twelve mice were administered i.p. 0.6% acetic acid (10 mL kg1, i.p.), and the
number of abdominal contractions was registered for 20 min, starting 30 min after the injection. Animals were pre-treated with
venlafaxine (10, 20, 40 and 60 mg kg1, i.p.), 30 min before acetic acid administration. DMSO 5% was used as vehicle (control
group). With the exception of 10 mg kg1, all other doses of venlafaxine (20, 40 and 60 mg kg1, i.p) produced significant
inhibition of writhing (p
Conclusions:
These results corroborate researche that reported the inhibitory effect of venlafaxine on the signs of spontaneous pain and
hindpaw mechanical and thermal hypersensitivity in animal models of tonic nociception and neuropathic pain. Studies continue
in an attempt to elucidate the action mechanisms of these effects.
Keywords: venlafaxina, inibidores da recaptao de serotonina e, dor, antidepressivos, analgesia
Financial Support: UFC
Resumo:17-198
DIFFERENTIAL REGULATION BY THALIDOMIDE OF SILICA-ACTIVATED MACROPHAGES IN VITRO
Santanna, E. S. ; Ciambarella, B. T. ; Pires, A. L. A. ; Cordeiro, R. S. B. ; Martins, M. A. ; Silva, P. M. R.

E.
Laboratory of Inflammation / Oswaldo Cruz Institute, FIOCRUZ
Objectives:
Silicosis is a chronic lung disease induced by the inhalation of crystalline silica. Prolonged deposition of particles in the alveoli or
bronchioles leads to lung inflammation and the formation of fibrotic scar tissue. Silica particles can activate different structural
and inflammatory cells including epithelial cells and macrophages. We previously showed that curative treatment with
thalidomide reduced lung function compromise and granuloma formation in experimental silicosis in mice. In this study we
aimed to investigate the potential direct effect of thalidomide on silica-induced macrophage activation in vitro.
Male Swiss-Webster mice (18 20g) were used. Cells were recovered from the peritoneal cavity after gently washing with RPMI
1640 complete media. Before particle exposure, cells were plated at 5 10 5 cells/cm2 on round glass coverslips in 24-well dishes
in RPMI 1640 and allowed to adhere for 1 h at 37C. The cells were then incubated with varying non-toxic concentrations of
silica (6.25 - 50 g/mL), for 6 h at 37C, and the dishes centrifuged at 1000 rpm for 10 min. The supernatant was recovered and
kept at -20oC for further analyses. We evaluated nitric oxide (NO) and IL-6 production by Greiss and ELISA techniques,
respectively. The cells were stained with Giemsa and phagocytosis was quantified under light microscope coupled to an image
analyzer system. The number of cells with internalized particles were counted and then divided by the total number of cells to
derive the percent phagocytosis. Survival rate was based on Trypan blue dye exclusion method. Macrophages were incubated
with different concentraitons of thalidomide (10-6 and 10-4 M), 1 hour before silica stimulation. All experimental procedures
were performed in accordance with the guidelines of the Committee on Use of Laboratory Animals of the Oswaldo Cruz
Foundation (L-034/09). Results: Exposure of macrophages with silica particles induced cell activation evidenced by the
morphological features and by the release of NO and IL-6. A concentration-dependent increase in the number of macrophages
with internalized particles was also detected. Pre-incubation of macrophages with thalidomide significantly reduced the levels of
IL-6 release, at the highest concentration, without affecting NO production. Interestingly, the phagocytosis rate increased under
condition of drug treatment. No cell death was noted under condition of silica stimulation or thalidomide treatment.
Conclusions:
Altogether our data show that thalidomide was able to differentially inhibit silica-induced chemical mediator production by
macrophages in vitro, under conditions where it potentiated phagocytosis. This indicates that cell activation and particle
internalization are not associated events. These findings also point out that macrophages seem to be potential pharmacological
targets for the anti-silicotic effect of thalidomide in mice.
Keywords: Silica particles, macrophages, IL-6, thalidomide
Financial Support: FIOCRUZ, CNPq, FAPERJ
Resumo:17-199
EFFECT OF NEW N-ACYLHYDRAZONES DERIVATIVES ANALOGOUS TO LASSBIO-294 ON
FORMALININDUCED HYPERNOCICEPTION IN MICE.
Silva, R. V. ; Veloso, R. R. ; Nogueira, M. C. O. ; Maia, R. C. ; Lima, L. M. ; Barreiro, E. J. ; Miranda, A.

L. P
Departamento de Frmacos Faculdade de Farmcia - LASSBio, UFRJ
Objectives:
The pharmacophoric group N-acylhydrazone has shown an important anti-inflammatory and analgesic activity. The compound
LASSBio-294 was identified as an important cardiotonic prototype also presenting anti-inflammatory and analgesic profile
(Quim. Nova 25; 1172, 2002). A previous screening study has demonstrated the antinociceptive potential of a new series of Nacylhydrazone (NAH) derivatives modified from LASSBio-294. Some of them (LASSBio-1499 e LASSBio-1532) showed an
antinociceptive potency of about 3 mol/kg (approx. 1 mg/kg) (SBFTE Congress, 2010). The aim of this study was to evaluate
this new series of NAH derivatives in inflammatory pain.
The antinociceptive activity was evaluated using the formalin-induced hipernociception test in mice. LASSBio derivatives (100
mol/kg; p.o.) and the standard AINE celecoxib were administered 1h before the intra-plantar injection of formalin 2.5% (20
l/paw). The time that mice spent licking or biting the injected paw was recorded and separated in two distinct periods: the first
period (earlier or neurogenic phase) was recorded 0-5 min after formalin injection and the second period (later or inflammatory
phase) was recorded 15-30 min after injection (Pain 51; 5, 1992). Results were expressed as the percentage of inhibition
compared to the vehicle control group (gum arabic 5%) (n = 6-12 animals; *p
Conclusions:
These findings suggest in part an anti-inflammatory profile for these compounds, since the hipernociception in the second phase
is a well characterized inflammatory response by the presence of neutrophils and of inflammatory mediators as TNF- and PGs,
which can also be modulated by the activation of the primary sensor neurons by formalin. The involvements of others
mechanisms of nociception as the activation of vanilloid (TRPV1, TRPA1) and glutamate receptors must be investigate.
Therefore, we highlight LASSBio-1476 as a potent and promising analgesic and anti-inflammatory compound useful for chronic
inflammatory diseases like arthritis.
Keywords: formalin, hypernociception, inflammation, N-acylhydrazone
Financial Support: CNPq, FAPERJ, PIBIC-UFRJ, INCT-Inofar.
Resumo:17-200
PHOSPHODIESTERASE-5 INHIBITION BY TADALAFIL PROVIDES TNF-DEPENDENT ANALGESIA IN
EXPERIMENTAL ARTHRITIS
Nunes, R. D. M. 1,1,1,1; Leite, A. C. R. D. M. 1; Rocha, F. A. C. D. 1

1
Departamento de Biomedicina/Faculdade de Medicina, UFC
2
3
Objectives:
We investigated the effect of the phosphodiesterase (PDE)-5 inhibitor tadalafil (TD) in the acute hypernociception in the
zymosan (Zy) and anterior cruciate ligament transection (ACLT) arthritis models.
Rats were subjected to either intra-articular (i.art.) injection of 1mg Zy or surgical ACLT, used as an osteoarthritis (OA) model.
Controls received saline i.art. or sham operation, respectively. Joint pain was evaluated using the articular incapacitation test
measured over 6 h following Zy or between 4 and 7 days after ACLT. Cell counts, tumor necrosis factor- (TNF), interleukin-1
(IL-1), and CINC-1 levels were measured in joint exudates 6 h after Zy. Groups received TD (0.02 0.5 mg kg-1 per os) or saline
2 h after i.art. Zy. Other groups received the -opioid receptor antagonist naloxone or the cGMP inhibitor 1H- [1,2,4] oxadiazolo
[4,3-a] quinoxalin-1-one (ODQ) prior to TD.Administration of TD dose-dependently inhibited hypernociception in Zy and OA
models. TD significantly decreased cell influx and TNF release in Zy arthritis while not altering IL-1 or CINC-1 levels. Pretreatment with ODQ but not with naloxone prevented the anti-inflammatory effects of TD.
Conclusions:
Therapeutic oral administration of TD provides analgesia mediated by guanylyl cyclase inhibition that is independent of
endogenous opioids release. This effect of TD is associated to a decrease in neutrophil influx and TNF release into inflamed
joints.
Keywords: Tadalafil, Phosphodiesterase, Arthritis, Pain, Cytokines
Financial Support: CNPq,CAPES
Resumo:17-201
EVALUATION OF THE PHARMACOLOGICAL PROPERTIES OF PARTITIONS OF THE CRUDE EXTRACT OF
MYRSINE CORIASEA
Carrasco, L. P. S. 1; Carvalho, M. F. 1; Leite, M. M. D. S. 1; Souza, P. A. D. 2; Cortes, W. S. 1

1
Dept de Cincias Fisiolgicas/Instituto de Biologia, UFRRJ
2
Instituto de Qumica, UFRJ
Objectives:
To evaluate the pharmacological properties of of the partitions ethyl acetate (EAE), hexane (EH)and buthanolic (EB) of crude
ethanol extract (EE) of Myrsine coriacea, in order to justify its potential use in human and veterinary medicine.
Antinociceptive activity was evaluated by the methods of formalin (J. Neurosci. Meth. 14, 69, 1985), peritonitis (Agents and
actions 32, 283, 1991) and pleurisy. In the method of formalin was investigated the mechanism of antinociceptive action. In this
method of solution 3% formalin is injected into the plantar hind paw of the hind paws of a mouse and observe the appearance of
two phases of the genesis of pain, phase 1 (F1) and 2 (F2) (F1, neurogenic pain and F2, inflammatory pain). We used the
following treatment groups: tap water, indomethacin (10 mg/kg) and the test group EB at a dose of 1 g/kg. The group treated with
EB showed no inhibition of F1 compared to control water. In the F2 this group inhibited by 56% (92.17 22.95 seconds, P
Conclusions:
Therefore, our results show an antinociceptive action of hexane, ethyl acetate and butanol partitions of M. coriacea. But there
was no central antinociceptive and anti-inflammatory mechanism seems to be involved in the antinociceptive action of these
extracts.
Keywords: antinflamatory, antinociception, Myrsine coriacea, pain
Financial Support: PIBIC/CNPq, DCF/IB/UFRRJ
Resumo:18-039
STUDY OF ANTIMICROBIAL AND ANTI-INFLAMMATORY ACTIVITY OF AN ANALOG OF PYRAZINAMIDE
Mendona, M. S. A. ; Canda, A. L. P. ; Lima, C. H. S. ; Souza, M. V. N. D. ; Henriques, M. G. M. O.

Laboratrio de Farmacologia Aplicada, Fiocruz
Objectives:
Tuberculosis is the most important cause of mortality due to a single infectious agent worldwide. The treatment for this infection
is based on drugs developed in the 60s. The pyrazinamide analogs synthesis has been an important field of research because of
its wide spectrum of pharmacological activities. In this context, the synthesis of pyrazinamide analogs has been an important area
of research, due to the broad spectrum of pharmacological activities. Recently we have selected by in vitro and in vivo pleurisy
studies three potential analogs, and among them the analogue 55 (A55) was selected for the experiments. The aim of this study
was to evaluate the anti-mycobacterial and immuno-modulatory activity of a pyrazinamide analog, A55, in macrophages infected
with BCG in vitro.
J774 cells (macrophages cell line) were infected for a period of 5 hours and subsequently treated with different concentrations of
the A55 and pyrazinamide (0.1 M, 0.01 M, 0.001 M, 0.0001 M, 0.00001 M) for 24 hours. The cytotoxicity of the analog was
analyzed by the Alamar blue method and the cell supernatant was collected determine nitrite concentration. Initially we observed
that the analog is cytotoxic for the cells when they are infected, in higher doses (0,001 M; 0,01 M;), while pyrazinamide
possessed little or no effect. This analogy in higher concentrations (0,001 M; 0,01 M; 0,1 M), can induce mycobacteria killing
(>20%). Regarding the release of nitric oxide in the supernatants, we observed that the analog was able to induce an increased
release (>40%) of this mediator in the highest concentration used (0.1 M), while pyrazinamide induced no change.
Conclusions:
Our results indicate that the analog has better in vitro antimicrobial profile in relation to pyrazinamide, due to its higher cytotoxic
activity to mycobacteria and increased production of nitric oxide.
Keywords: Tuberculosis, Pyrazinamide, Inflamation, Mycobacteria
Financial Support: PIBITI/Fiocruz, CNPq, FAPERJ, Farmanguinhos
Resumo:18-040
IN VITRO EFFECTS OF HYDROCORTISONE AND CYCLOSPORINE A IN THE DEVELOPMENT OF MURINE
THYMOCYTES
Costa, K. M. 1; Paiva, L. S. 2; Rumjanek, V. M. 1

1
2
Departamento de Imunobiologia/Instituto de Biologia, UFF
Objectives:
P-glycoprotein is a transmembrane protein involved in the transport of several lipophilic compounds. Although this glycoprotein
was first described in the phenomenon of multidrug resistance, its expression also occurs in normal cells, as in thymocytes. The
expression of P-glycoprotein in these cells seems to be regulated during development and the subpopulation that does not show
activity related to P-glycoprotein is the most sensitive to exposure to glucocorticoids. Furthermore, corticoids are P-glycoprotein
substrates. This suggested the involvement of this transporter protein on the control of intracellular levels of glucocorticoids.
There are evidences that glucocorticoids play a regulatory role on the thymus, more specifically, on the thymic selection. To
understand the role of this glycoprotein on susceptibility to glucocorticoids, we treated thymocytes with hydrocortisone, a
glucocorticoid, and cyclosporine A, a known inhibitor of P-glycoprotein and evaluated the amount of viable cells.
The suspension of thymocytes extracted from C57BL/6 females mice of 4-6 weeks old or 3-5 months old were treated with
hydrocortisone (0.01 - 0.1 M) and / or cyclosporine A (0.2 or 0.4 g/ml) for 17-18 hours. After treatment, we counted the
number of viable cells by Trypan Blue dye exclusion. For analysis of P-glycoprotein activity, cells were incubated with 0.2 g/ml
rhodamine 123, fluorescent substrate of P-glycoprotein, in the presence or absence of 0.4 g/ml of cyclosporine A for 30 minutes.
The cells were washed and incubated with medium in the presence or absence of cyclosporine A for 30 minutes. After that time,
cells were stained with anti-mouse CD4/CD8 and analyzed by flow cytometry. The in vitro treatment with hydrocortisone caused
a dose-dependent decrease in the amount of thymocytes (n=3). The most immature subpopulations were more sensitive to
treatment, whereas mature cells were more resistant. Treatment with cyclosporine A showed a tendency to dose-dependent
reduction in the number of viable thymocytes (n=3). The effect of treatment with hydrocortisone and cyclosporine A was
predominantly additive only in CD4-CD8- and CD4+ CD8+ cells from mice of 4-6 weeks old (n=5). In the other subpopulations
and in mice of 3-5 months old, there were no significant differences between treatment with hydrocortisone alone and in the
presence of cyclosporine A. Finally, we found that treatment with hydrocortisone did not increase the percentage of cells with Pglycoprotein activity compared to control (n=6), demonstrating that the activity of this glycoprotein did not affect treatment with
glucocorticoid.
Conclusions:
Treatment with hydrocortisone reduced the amount of cells independent of P-glycoprotein activity, suggesting that susceptibility
of thymocytes to glucocorticoid exposure is not directly related to the activity of this glycoprotein.
Keywords: Thymocytes, Glucocorticoids, P-glycoprotein, Cyclosporine A
Financial Support: CNPq, FAPERJ.
Resumo:18-041
EVALUATION OF LIPID AND PROTEIN OXIDATION AND BUTYRYLCHOLINESTERASE ACTIVITY IN
SERUM OF PATIENTS WITH INDETERMINATE FORM OF CHAGAS DISEASE
Schlemmer, J. B. ; Souza, V. D. C. G. ; Thorstenberg, M. L. ; Castilhos, L. G. ; Rocha, B. C. ; Schlemmer,

K. B. ; Bertoldo, T. M. D. ; Gonalves, J. F. ; Costa, P. ; Leal, D. B. R.
Departamento de Microbiologia e Parasitologia, UFSM
Objectives:
Aim: Chagas disease is caused by Trypanosoma cruzi and is characterized by a chronic inflammation. It is known that changes
in the redox state and regulation of inflammatory process are associated with the progress of several chronic inflammations such
as Chagas disease. Therefore, the oxidative profile determined by the lipid peroxidation, protein carbonylation and
butyrylcholinesterase (BuChE) enzyme activity were determined in the serum of patients with indeterminate form of Chagas
disease.
Methods and Results: This study was performed in a control group constituted by 22 T. cruzi serum-negative subjects with an
average age of 47 years old and an infected group constituted by 22 patients with indetermined form of Chagas disease (IFCD)
with an average age of 54 years old, selected from the University Hospital of Santa Maria. The protocol was approved by the
Human Ethics Committee from the Federal University of Santa Maria under the number 243/2009 and all the subjects gave
written consent. The lipid peroxidation was determined in serum samples by the thiobarbituric acid method (Free Radic Biol
Med. 20:251-256,1996) with the formation of malondialdehyde (MDA) which it was readed at 532 nm. The results were
expressed as nmoles MDA/mL of serum. The carbonylation of serum proteins was determined by addition of 2,4-dinitrophenylhydrazine to serum as previously standardized (Methods Enzym 186:464-478). The proteins were precipited by 10%
trichloroacetic acid and incubated at 37C in phosphate buffer, pH 8.0. After this, the color intensity of the supernatant was
measured at 370 nm and the carbonyl content was expressed as nmol/mg proteins. The BuChE enzymatic assay was determined
in serum by a modification of the spectrophotometric method (Biochem. Pharmacol. 7:88-95, 1961). The method is based on the
formation of the yellow anion, 5,5-dithio-bis-acid-nitrobenzoic, measured by absorbance at 412 nm during 2-min incubation at
25C. The enzyme activity was expressed in mol BuSCh/h/mg of protein. All assays were run in duplicate and the data were
analyzed statistically by the Students t test. Differences were considered significant when the probability was P<0.05).
Conclusions:
Conclusion: Given the results of TBARS and protein carbonyl, we can conclude that the Chagas disease induces a redox
imbalance which leads to alterations in cell function by oxidation and damage of macromolecules, membranes, proteins and
DNA. These results can be explained by a possible ineffective antioxidant response and also because Chagas disease develops
an inflammatory process, with the formation of reactive oxygen (ROS), mediated by cytotoxic reactions by the immune system.
The decreased of BuChE activity in the IFCD group suggests an increase in the serum levels of the acetylcholine in this group,
possibly in response to inflammatory damages triggered by the invasion of Trypanosoma cruzi in several tissues, since the
acetylcholine has anti-inflammatory properties.
Keywords: Butyrylcholinesterase, Chagas' Disease, Lipid Oxidation, Protein Oxidation
Financial Support: FIPE, CNPq, CAPES
Resumo:18-042
INTERACTION OF MYCOBACTERIUM LEPRAE WITH HUMAN MACROPHAGES AND DENDRITIC CELLS:
ROLE OF NITRIC OXIDE AND EXPRESSION OF SURFACE MOLECULES.
Souza, I. C. C. ; Castro, H. C. ; Santos, D. O.

Departamento de Biologia Celular e Molecular, UFF
Objectives:
Leprosy is a chronic infectious disease caused by Mycobacterium leprae . The clinical manifestations depend on the patient's
immune response to the bacillus. These findings suggested that differences between the tuberculoid and lepromatous leprosy are
correlated with the presence of different profiles of T cells. However, determining the profile of T cells (Th1 or Th2) primarily
depends on the nature of the antigen-presenting cell (APC), as already demonstrated by our group, demonstrating the reversal of
the inefficiency of the cellular immune response against M. leprae in lepromatous leprosy when dendritic cells (DCs) were used
as APCs.(Clin Exp Derm 32(1):75, 2006). Nitric oxide (NO) and its reactive intermediates (RNI) are microbicides, also
characterizing it as a mediator in the defense against infections. The aim of this study was to investigate the interaction of M.
leprae to macrophages and dendritic cells derived from monocytes isolated from human peripheral blood using the nitric oxide
production and expression of surface molecules as parameters.
This project is registered in the Ethics Committee of HUCFF (CAAE 0157.0.197.000-09). Mononuclear cells were purified from
peripheral blood of healthy donors and then monocytes were isolates by cold aggregation (J Leprosy 70:15, 2001). Monocytes
were cultured in labteks (2x105/ml) in the presence or not of rhIL-4 (1000U/mL, Innogenetics, Belgium) and rhGM-CSF
(100U/mL, Schering-Plough, Belgium) for 7 days, to differentiate in dendritic cells of macrophages, respectively. After
differentiation, and inhibitor of nitric oxide synthase was additioned (L-NAME 20M) for 1 hour. After that, cells were infected
with M. leprae (20g/mL, Colorado University, USA) or BCG (FIOCRUZ). Cells were fixed with paraformaldehyde and stained
by Kynioun method. Part of the cultures was destinated to flow citometry. By light microscopy, we observed a large increase in
infection by M. leprae in the presence of L-NAME, both in macrophages and in dendritic cells, suggesting that nitric oxide is
essential for cell defense against infection. By flow cytometry we have seen an increase in the expression of CD209 when
macrophages are infected by M. leprae , which together with the absence of NO and RNI, facilitate the entry of M. leprae in the
cell. When dendritic cells were stimulated with BCG, there was a significant increase in the expression of CD209 compared to
the macrophages, confirming the importance of DCs in the construction of an effective immune response.
Conclusions:
By light microscopy, we could conclude that macrophages are more likely to be infected by M. leprae than dendritic cells, and
the inhibition of nitric oxide synthesis allows the entry and permanency of M. leprae and BCG inside the cells, macrophages or
dendritic cells, caractherizing nitric oxide as a important mediator in combating infections. By flow citometry, we observed that
M. leprae induce DC-SIGN expression in macrophages, as observed by the CD209 marking.
Keywords: dendritic cells, leprosy, nitric oxide, Mycobacterium leprae
Financial Support: CNPq, FAPERJ, UFF
Resumo:18-043
ASCARIS SUUM EXPERIMENTAL INFECTION MODULATES ACUTE INFLAMMATION VIA
CD4+CD25+FOXP3+ T CELLS.
Titz, T. D. O. ; Macedo-soares M. F; Araujo C. A. A

Instituto Butantan, IBUT
Objectives:
The aim of this study was to investigated the expansion of T cell clones by Ascaris suum infection and the effect of these cells on
LPS-induced acute inflammation
Methods: BALB/c mice weighting about 22 g were infected by intragastric route with 2500 Ascaris suum embryonated eggs. A
week later the mesenteric lymph node cells were recovered. The expression of CD4+CD25+FoxP3+ markers in lymph node cells
from non-infected and Ascaris-infected mice was detected using PE-Cy5.5-anti-CD25, FITC-anti-CD4 antibodies and PE-antiFoxP3 antibodies. All samples were acquired using FACSCalibur and analyzed by FlowJo software. The TCD4+CD25- and
TCD4+CD25+ cell population from infected or non-infected mice were purified from lymph node cells by magnetic activatedcell sorter (MACS). These cells were adoptively transferred to recipient mice by intravenous route and 3 days later the air
pouches induced by sterile air injections on the back of the recipient mice were LPS-injected. Control group received only PBS in
their pouches. Three hours after the stimulation, the air pouche exsudates were recovered to the measurement of cytokines levels
by ELISA and quantification of cellular migration. Results: FACS analyses showed the percentage of T cell subpopulations from
non-infected and Ascaris suum-infected mice. The total CD4+ T cell numbers did not differ in Ascaris suum-primed and nonprimed total cells. The number of CD25+FoxP3+ cells gated on CD4+ cell population presented a 4 fold increase in Ascarisprimed total cells compared to non-primed cells. Furthermore, CD4+CD25-FoxP3+ cell numbers were also higher in Ascarisprimed total cells (14.1%) compared to non-primed cells (4.8%). Regarding the suppressive effect of these cell clones our results
demonstrated that adoptive transfer of CD4+CD25+T cells from infected mice resulted in decreasing of leukocyte recruitment
(3.5 fold) and inflammatory cytokines levels (IL-1, IL-6 and TNF-), but increasing of IL-10 and TGF- cytokines. On the
contrary, transfer of TCD4+CD25- cells or TCD4+CD25+ from non-infected mice had no effect on leukocyte influx or cytokines
levels into the air pouches compared to the positive control group
Conclusions:
In our previous studies, the regulatory cytokine IL-10 have been implicated in the down-regulatory mechanisms induced by
Ascaris suum. The results obtained herein indicate that the source of these cytokines may be CD4+CD25+ T cells. Thus, when
these cells, but not other T cell types, were obtained from lymph nodes of Ascaris-infected mice and adoptively transferred to
recipient mice, they prevented the inflammatory response induced by LPS. In conclusion the present study shows that Ascaris
suum infection expands the CD4+CD25+FoxP3+ T cells clones producing IL-10, which are responsible for the anti-inflammatory
activity.
Keywords: Ascaris suum, Infection, immunemodulation
Financial Support: Financial support: CNPq and FAPESP
Resumo:18-044
OLEIC ACID SUPLEMENTATION ATTENUATES INFLAMMATION AFTER A CLP-INDUCED SEPSIS IN SWISS
MICE
Medeiros-de-moraes, I. M. ; Campbell, C. ; Magno, F; Arajo, C. V ; Bozza, P. T. ; Gonalves-dealbuquerque, C. F; Silva, A. R. ; Castro-faria-neto, H. C

laboratrio de Imunofarmacologia/FIOCRUZ, FIOCRUZ
Objectives:
sepsis accounts for a huge number of deaths in intensive care units worldwide. Sepsis describes a complex clinical syndrome that
results from an infection, setting off a cascade of systemic inflammatory responses that can lead to multiple organ failure and
death. Leite et al has showed that mice fed for 6 weeks with an olive oil diet were resistant to endotoxic shock, with 60% survival
at 168 h (Shock 23; 173, 2005). Olive oil is composed by different polyunsaturated fatty acids such as omega 3 and 6 but the
monounsaturated fatty acid omega 9, also known as oleic acid (OA), that is the main component of olive oil - highly consumed in
Mediterranean diet. We aim to investigate the role of OA in an experimental model of sepsis.
Swiss mice were given daily doses (orally) of OA, at 282 g/animal, for 14 days. Control animals received saline. In the sixteenth
day polymicrobial sepsis was induced by cecal ligation and puncture (CLP). Immediately after the procedure all mice received
volemic reposition and after 6 h animals were given imipenem. 24 h after surgery mice were euthanized and the peritoneal cavity
was rinsed with sterile saline. Total leukocyte counts were performed in a Neubauer chamber and differential leukocyte analysis
done in cytosmears stained with May-Grunwald Giemsa. The supernatant and serum were collected for cytokine quantification
and biochemical analysis. In another set of experiments survival rate was determined daily for 7 days in separate groups of at
least 10 animals for each condition. Mice fed with OA were resistant to sepsis, with a 90% survival rate at 168 h compared to
saline treated mice (58%). OA supplementation in CLP-subject animals led to a significantly decrease in the total leukocyte
counts (32.33 x 106 3.93 - mean SEM), mainly neutrophils, compared to mice that received saline (46.10 x 106 2.84).
However, in mice that consumed OA the levels of TNF, IL-10, IL-6 and PGE2 were not significantly different from mice fed
with saline submitted to CLP. Moreover, mice fed with OA had a significantly lower level of bacteria (13.6 x 106) in the
peritoneal lavage leukocyte compared to mice submitted to CLP (59.6 x 106). The hepatic and renal functions were assessed by
quantifying the serum levels of aspartate transaminase and creatinine.
Conclusions:
our data suggests that pretreatment with OA reduces mortality in an experimental model of sepsis possibly through immune
response modulation. One mechanism involved may be due to increased bacterial clearance in mice fed with OA. More data are
required to clarify this mechanism of increased survival.

Keywords: oleic acid, sepsis, inflammation
Financial Support: CNPq, FIOCRUZ and FAPERJ
Resumo:18-045
LEUKOTRIENE B4/BLT1 MEDIATES T LYMPHOCYTE MIGRATION DURING INFLAMMATION
Souza-martins, R. 1; Costa, M. F. S. 1; Souza, M. C. 1; Piva, B. 2; Diaz, B. L. 2; Peters-golden, M. 3;

Henriques, M. G. M. O. 1; Canetti, C. A. 2; Penido, C. 1
1
Fundao Oswaldo Cruz - Farmanguinhos, FIOCRUZ
2
Lab. de Inflamao - Universidade Federal do Rio de Janeiro, UFRJ
3
University of Michigan Health System , UMHS
Objectives:
T lymphocytes are unconventional T cells that comprise a minor subset of T cells in lymphoid organs, which are preferentially
distributed in peripheral tissues. These cells exert an early pro-inflammatory role followed by a subsequent regulatory role in an
attempt to restrain the inflammatory response. T lymphocytes increase in number at inflammatory sites during infection and
allergy, a phenomenon mediated via migration towards chemotactic factors and/or local proliferation. In this context, the aim of
this project is to investigate the involvement of the 5-lipoxygenase (5-LO)-derived lipid mediator leukotriene (LT)B4 in T cell
migration and activation.
Pleurisy was induced by an intrapleural (i.pl.) injection of LTB4 (0.5 g/cavity), LPS (250 ng/cavity), OVA (12.5 g/cavity), or
Mycobacterium bovis BCG (4x105 CFU/cavity). OVA challenge was induced in mice 14 days after prior sensitization by a
subcutaneous injection of 200 l of a mixture of OVA (50 g) and aluminum hydroxide (5 mg). Pleural cells were recovered
from thoracic cavities after washing with 500 l of PBS containing EDTA (10 mM, pH 7.4). The i.pl. injection of LPS triggered
increased levels of LTB4 in C57BL/6 mouse pleural cavities (saline: 12,27+13,56; LPS: 47,74+6,18 pg/mL). The in vivo
inhibition of 5-lipoxygenase (5-LO) activity by zileuton, impaired LPS-induced T cell accumulation into pleural cavities
(saline: 6,33+0,75; LPS:12,14+1,18; zileuton: 8,00+0,72 x 103 cells/cavity). Accordingly, 5-LO knockout (5-LO-/-) mice failed
to recruit T cells into the inflammatory site after LPS i.pl. stimulation (WT-saline: 4,11+1,31; WT-LPS: 19,87+4,19; 5-LO-/- saline: 4,19+1,27; 5-LO-/--LPS: 6,42+1,81 x 103 cells/cavity). The antagonism of high affinity LTB4 receptor, BLT1, by
CP105,696 was also capable to impair T cell accumulation in mouse pleural cavities induced by LPS (saline: 2,45+1,11; LPS:
10,72+0,87; CP105,696: 7,01+0,53 x 103 cells/cavity). Confirming that T lymphocytes can directly respond to LTB4, we
demonstrate that nave resident T lymphocytes, as well as LPS recruited T cells express BLT1 receptor and isolated T
cells were found to undergo actin cytoskeleton reorganization when incubated with LTB4 in vitro. LTB4/BLT1 also accounted
for T cell migration induced by BCG (saline: 5,53+0,857; BCG: 22,04+5,47; CP105,696: 6,82+1,33 x 103 cells/cavity) or
during antigen challenge in sensitized mice (saline: 3,51+0,98; OVA: 29,42+3,16; CP105,696: 13,29+4,79 x 103 cells/cavity).
Conclusions:
These data show that LTB4/BLT1 pathway mediates T cell migration into the inflammed tissue in response to diverse stimuli.
Keywords: T lymphocytes, Leucotriene B4, CCL2/MCP-1, cell migration, cell activation
Financial Support: CNPq, FAPERJ, FIOCRUZ.
Resumo:18-046
PROLIFERATIVE ACTION INDUCED BY ONIL, A NOVEL LECTIN PURIFIED FROM TILAPIA FISH
Silva, C. D. C. 1; Coriolano, M. C. 1; Melo, C. M. L. D. 3; Silva, M. A. L. D. 1; Carvalho, E. V. M. M. D. 1;

Bezerra, R. D. S. 1; Santos, A. J. G. D. 7; Pereira, V. R. A. 3; Coelho, L. C. B. B. 1
1
Depart. de Bioqumica/UFPE, UFPE
3
Centro de Pesquisas Aggeu Magalhes (CPqAM/Fiocruz), Fiocruz
7
DEPAq/UFRPE, UFRPE
Objectives:
This in vitro study analyzed cellular proliferation and cytokine production in mice splenocytes cultures treated with OniL, a lectin
purified from Oreochromis niloticus serum.
Methods: Experimental assays used mice (BALB/c, male, 30 days old, 5/group); all animals were killed and treated in
accordance with the guidelines of the Oswaldo Cruz Foundation Commission for Experiments (Rio de Janeiro, Brazil) with
Laboratory Animals (Ministry of Health, Brazil, 0266/05). OniL is a lectin from serum of tilapia fish (Oreochromis niloticus)
purified by affinity chromatography on Concanavalin A-Sepharose 4B (Sigma). Cancanavalia ensiformis (Concanavalin A Con
A) was purchased from Sigma Chemical Co. Cellular proliferation assay was performed by [3H]-thymidine incorporation. Cells
of each group were in vitro treated for 24 h with Con A (2.5 g/mL) and OniL (2.5, 5 and 10 g/mL) to evaluate the proliferative
activity of lectins. The unstimulated culture plate was used as a negative control. SI values 3 were considered representative of
positive proliferation. Cytokine production was measured by sandwich ELISA and IL-2 and IL-6 cytokines were quantified. Each
well received Con A (2.5 g/mL) or OniL (10 g/mL), and supernatants from cultures stimulated in vitro with lectins were
obtained at 24, 48, 72 h and 6 days. Cells kept in culture medium were also obtained as a negative control. Cell viability assay
was performed by Annexin V conjugated with fluorescein isothiocyanate and propidium iodide. Splenocytes (106 cells) were
stimulated with lectins. Annexin-FITC-PI+ cells were considered necrotic cells and Annexin-FITC+PI-represented splenocytes
in the early stage of apoptosis. Nonparametric assays were used in statistical analysis. All results were expressed as mean values
of groups standard deviation and analyzed considering the value of P < 0.05 as statistically significant. Results: A great number
of lectins with distinct characteristics and specificities have been used for their immunomodulatory and proliferative activities,
but few studies show immunological profile induced by fish lectins. Higher and statistical indices of proliferation were induced
by OniL in relation to control cells. OniL showed higher mitogenic activity in relation to control and Con A for all analyzed
concentrations. Higher IL-2 and IL-6 production was promoted by OniL; sometimes this production was also superior to Con A.
Results of viability assay showed that OniL did not promote cell damage in neither analyzed protocols. Beyond, Con A induced
higher apoptosis and necrosis in both 24 and 48 h in relation to control and OniL cultures.
Conclusions:
OniL induces higher proliferative response; this lectin can be used as a mitogenic agent in immunostimulatory assays
Keywords: LECTIN, TILAPIA, PROLIFERATIVE ACTION, IMMUNOMODULATORY, CYTOKINE
Financial Support: CNPq, FACEPE
Resumo:18-047
LECTIN OF COBIA SERUM, RCAL, PROMOTED MITOGENIC RESPONSE IN MICE BALB/C SPLENOCYTES
Coriolano, M. C. 1; Silva, C. D. C. D. 1; Melo, C. M. L. 2; Bezerra, R. S. 1; Santos, A. J. G. 3; Pereira, V. R.

A. 2; Coelho, L. C. B. B. 1
1
DBIOq/UFPE, UFPE
2
CPqAM/FIOCRUZ, FIOCRUZ
3
DEPAq/UFRPE, UFRPE
Objectives:
Aim: The aim of this study was to evaluate the proliferative response and cytokine production in mice splenocytes in vitro
stimulated with a novel Rachycentron canadum lectin of mannose recognition, RcaL.
Methods:Experimental assays utilized mice (BALB/c, male, 30 days old, 5/group). All mice were killed and treated in
accordance with the guidelines of the Oswaldo Cruz Foundation Commission for Experiments (Rio de Janeiro, Brazil) with
Laboratory Animals (Ministry of Health, Brazil, 0266/05). RcaL is a lectin isolated and purified from serum of cobia fish (R.
canadum) obtained by affinity chromatography on Concanavalin A-Sepharose 4B (Sigma). Canavalia ensiformis (Concanavalin
A Con A) was purchased from Sigma Chemical Co. Cellular proliferation assay was performed by [3H]-thymidine
incorporation. Cells of each group were treated, in vitro, for 24 h with Con A (2.5 g/mL) and RcaL (2.5, 5 and 10 g/mL) to
evaluate the proliferative activity of these lectins. The unstimulated culture plate was used as a negative control. SI values 3
were considered representative of positive proliferation. Cytokine production was measured by sandwich ELISA; IL-2 and IL-6
cytokines were quantified. Each well received Con A (2.5 g/mL) or RcaL (10 g/mL) lectins, and supernatants from cultures
stimulated in vitro with lectins were obtained at 24, 48, 72 h and 6 days. Cells maintained in culture medium were also obtained
as a negative control. Cell viability assay was performed by Annexin V conjugated with fluorescein isothiocyanate and propidium
iodide. Splenocytes (106 cells) were stimulated with lectins. Annexin-FITC-PI+ cells were considered necrotic cells and
Annexin-FITC+PI-represented splenocytes in the early stage of apoptosis. Nonparametric tests were used in statistical analysis.
All results were expressed as mean values of groups standard deviation and analysed considering the value of P < 0.05 as
statistically significant. Results: Mannose binding lectins are known by their immunostimulatory properties, such as cell
proliferation and cytokine production. Higher and statistical indices of proliferation were induced by RcaL in relation to control
cells. Although Con A stimulus promoted statistical values of proliferation in relation to control the stimulation was lower in
relation to RcaL. Furthermore, RcaL and Con A induced higher IL-2 and IL-6 production; results showed similar behavior for
both lectins. In contrast, RcaL and Con A showed similar behavior and induced higher IL-6 production. Only late apoptosis was
promoted by RcaL treatment and it also induced higher necrosis in relation to Con A.
Conclusions:
Conclusions: Results showed that RcaL induces proliferative response and suggested that this lectin can be used as a mitogenic
agent in immunostimulatory assays.
Keywords: CELLULAR PROLIFERATION, COBIA, CYTOKINE, IMMUNOSTIMULATORY PROPERTIES, LECTIN
Financial Support: FACEPE, CNPq.
Resumo:18-048
TSPO LIGAND MODIFY ADHESION MOLECULE EXPRESSION DUE TO BLOCKAGE OF CALCIUM INFLUX
INDUCED BY FMLP
de Lima, C. B. 1; Tamura, E. K. 3; Markus, R. P. 3; Neto, J. P. 2; Farsky, S. H. P. 1

1
Laboratory of Experimental Toxicology, FCF-USP
2
Laboratory of Applied Pharmacology and Toxicology, FMVZ-USP
3
Department of Physiology, IB-USP
Objectives:
Benzodiazepines (BZD) are one of the most commonly used groups of anxiolytic drugs, whose effects are mediated through
binding on neuronal post-synaptic plasma membrane GAGAA receptors. However, a second class of benzodiazepine binding
sites, translocator protein (TSPO), has been found in many peripheral tissues and cell populations, as immune and endothelial
cells. Studies have been shown that TSPO may be related, directly or indirectly, to the modulation of BZD on immune system.
Therefore, these mechanisms involved in the actions of BZD via TSPO are not fully comprehended. This worked investigated
TSPO-binding drugs effects on leukocyte-endothelial interactions, on adhesion molecules expressions and calcium influx.
Intravital microscopy assays were performed in the mesentery microcirculation of male Wistar rats (50-60 days old) after topical
application of TSPO ligands (PK 11195 or Ro 5-4864; 100nM) and fMLP (10-8M; topical application). In other set of assay,
circulating leukocytes were in vitro incubated with TSPO ligands (100nM) and 1 hour after incubated with fMLP (10-8M) to
quantify adhesion molecules expressions by flow cytometry. To evaluate calcium influx by confocal microscopy, isolated
neutrophils were incubated with Fluo-3AM previously to TSPO ligands (100nM) and fMLP (10-8M) application All procedures
were performed according to protocols approved by the Brazilian Society of Science of Laboratory Animals for proper care and
use of experimental animals. fMLP stimulation decreased rolling cells and increased adherent cells in control animals, and Ro54864 pre-treatment abolished these effects. In vitro fMLP stimulation reduced and enhanced L-selectin and &beta2-integrin
expression on neutrophils, respectively, and Ro5-4864 treatment inhibited the reduction on L-selectin expression caused by
fMLP. fMLP in vitro stimulation enhanced PECAM-1 expression, but Ro5-4864 abolished this effect. Ro5-4864 inhibited and
PK11195 enhanced maximal response and AUC of calcium influx induced by fMLP.
Conclusions:
Results show that TSPO ligand alters in vivo leukocyte-endothelial interactions evoked by fMLP and Ro5-4864, but not
PK11195, blocks the fMLP effect on neutrophil L-selectin and PECAM-1 expression which may be dependent on inhibition of
calcium influx.
Keywords: adhesion molecules, calcium, TSPO
Financial Support: FAPESP 09/52245-1
Resumo:18-049
PRODUCTS DERIVED FROM LEUKEMIC CELLS MODULATE THE GENERATION OF DENDRITIC CELLS IN
DIFFERENT STAGES
Motta, J. M. ; Rumjanek, V. M.
Objectives:
Dendritic cells (DCs) are professional antigen-presenting cells, specialized in activating T lymphocytes. DCs play an important
role in controlling tumors, since they are able to recognize and capture tumor antigens, initiating an antitumoral immune
response. However, tumor cells have developed mechanisms to inhibit immune responses, favoring tumor progression. Some
products secreted by tumor cells present immunosuppressive characteristics and they can affect different kinds of immune cells,
including DCs. There are many studies involving products released by solid tumors and how they modulate DCs, but there is a
paucity of information about products released by leukemic cells. In this case, leukemic cell factors are secreted in the blood,
where monocytes (DCs precursors) are present; therefore they could present alterations in their development. To investigate this
hypothesis, we decided to analyze a possible influence of leukemic cell products on the development and function of DCs in
different stages.
For this, the tumor cell line used was K562, derived from chronic myeloid leukemia. Monocytes, obtained from healthy donors,
were separated by Ficoll-Hystopaque density gradient and cultured in the presence of IL-4 and GM-CSF, stimuli for DC
differentiation, and also in the presence of K562 supernatant. After 5 days, CD14, CD1a expression and dextran phagocytosis
were evaluated in these cells by flow cytometry. In another experiment, looking at the effect of tumor products on DC activation,
immature DCs were cultured with TNF-alpha for a further 2 days. After this period, CD83 expression by DCs was measured by
flow cytometry. Moreover, activated DCs were co-cultured with lymphocytes obtained from a second donor for 24 hours and
lymphocytes expression of CD69 was evaluated. During a control differentiation, monocytes down regulate CD14 expression
and start to express CD1a. In this stage, immature DCs present high phagocytic capacity. In the presence of tumor supernatant,
CD14 expression remained high and CD1a expression was low (N=6). However, the addition of tumor supernatant did not
interfere with dextran phagocytosis by monocytes stimulated to differentiate into DCs (N=4). If activated, DCs considerably
increase CD83 expression and acquire high ability to activate lymphocytes. It was observed that monocytes differentiated in the
presence of tumor supernatant and then stimulated to activation expressed less CD83 (N=3). Moreover, tumor products appear to
inhibit the appearance of CD69 expression on lymphocytes co-cultured with DCs differentiated with tumor supernatants (N=2).
Conclusions:
Finally, these results suggest that soluble products released by leukemic cells affect DC differentiation and activation, suggesting
that, in this condition, these cells become unable to complete their development.
Keywords: dendritic cells, differentiation, leukemic cell products
Resumo:18-050
MODULATION OF MONOCYTE SURFACE RECEPTORS BY OUABAIN
Teixeira; M. P. C. ; Valente; R. C. ; Rumjanek; V. M.

INSTITUTO DE BIOQUMICA MDICA, UFRJ
Objectives:
Ouabain is a steroid produced by hypothalamus and adrenal glands that is capable of binding to and inhibiting Na+K+ATPase.
Also, ouabain is able to trigger different cell signaling pathways. Regarding the immune system, ouabain was shown to modulate
pro-inflammatory cytokines release by mononuclear cells, inhibit mitogen-induced proliferation of lymphocytes and, augment
intracellular calcium levels and CD69 expression also in lymphocytes. However, ouabain effects on monocytes, especially human
monocytes, are still poorly understood. Thus, the aim of the present work was to examine if ouabain was able to modulate the
expression of different surface molecules involved in the activation and effector functions of monocytes.
Mononuclear cells from healthy volunteers were separated by Ficoll-Hystopaque density gradient and cultured for 24 hours in the
presence or absence of 10-7M ouabain. After the incubation period, the expression of CD80, CD86, CD40 and CD69 molecules
were assessed using flow cytometry. Our results show that ouabain induced an increase in the expression of CD86, CD80 and
CD69 molecules, although, led only to a small increase of CD40 (n=4). When ouabain is added at the same time as LPS (antigen
capable of activating monocytes), ouabain seems to block LPS-induced CD40 augmented expression (n=5). On the other hand, in
the case of CD69, co-incubation with ouabain and LPS promoted an even higher increase in the expression of this molecule than
the incubation with ouabain or LPS alone (n=8).
Conclusions:
Together, these results showed that ouabain, in the presence or absence of LPS, is able to modulate the expression of different
surface molecules related to the state of activation and function of monocytes, suggesting an immunomodulatory role for ouabain
on these cells.
Keywords: Monocytes, Ouabain, Surface molecules
Financial Support: CAPES, CNPq and FAPERJ.
Resumo:18-051
EVALUATION OF THE PROFILE OF SERUM GLYCOPROTEINS OF HEALTHY INDIVIDUALS AND PATIENTS
WITH DENGUE FEVER AND DENGUE HEMORRHAGIC FEVER USING PURE LECTINS
Melo, M. S. ; Aranda-souza, M. A. ; Paula, R. A. ; Correia, M. T. S.

Departamento de Bioqumica, UFPE
Objectives:
Lectins are proteins or glycoproteins capable of interacting reversibly with carbohydrates through at least two binding sites, plant
cells coalesce and / or animals and precipitate glycoconjugates. The specificity of the interaction between lectins to
glycoconjugates free in solution or present on the cell surface makes these biological properties of biomolecules such as
differential recognition of carbohydrates or glycoconjugates expressed in normal or transformed tissues.The aim of this study was
to use lectins BmoLL, Cramoll-1, 4 and CrataBL for characterization of profile serum glycoproteins of healthy individuals (IH)
and patients with dengue fever (DF) and dengue hemorrhagic fever (DHF).
Lectins of seed Cratylia mollis (Cramoll-1,4 specific for D-glucose), leaf Bauhinia monandra (BmoLL specific for D-galactose)
and the bark of Crataeva tapia (CrataBL, specific glycoproteins) were purified. Radial diffusion assay was performed using 1%
(w/v) agarose gel containing 0.15M-NaCl, in Petri dishes. Serum samples (20L) IH, DF and DHF were applied in a circular
distribution around a central pit containing a lectin (20L) studied. Diffusion was allowed to take place for 48 h at 4C in
humidity chambers. The gels were washed with 0.15M-NaCl and distilled water, dried and stained with 0,005% (w/v) Coomassie
Brilliant Blue. The radial diffusion assay in agarose gel showed lines of precipitation for the lectins Cramoll-1,4, BmoLL and
CrataBL showing the recognition and binding of lectins to carbohydrates of glycoproteins of serum samples, justifying the use of
lectins to obtain glycoproteins that can differentiate pathologies in serum from healthy individuals or disease.
Conclusions:
The lectins tested (BmoLL, Cramoll-1,4 and CrataBL) had the potential to promote the isolation of glycoproteins in serum from
healthy individuals and patients with dengue fever or dengue hemorrhagic fever.
Keywords: dengue, glycoprotein, lectin
Financial Support: CAPES, CNPq, FACEPE
Resumo:18-052
LTB4 AS CHEMOATTRACTANT FACTOR IN THE REGULATORY T CELLS MIGRATION
Pecli, C. S. 1; Molinaro, R 5; Peters-golden, M. 2; Kunkel, S. L. 3; Canetti, C. . A. 4; Benjamim, C. F. 1

1
Instituto Cincias Biomdicas, UFRJ
2
Division of Pulmonary and Critical Care Medicine, U M
3
Department of Pathology, U M
4
5
Objectives:
Leukotrienes are lipidic mediators, among these the leukotriene B4 (LTB4) plays a critical role in leukocyte recruitment and
activation. LTB4 is derived from the metabolism of arachidonic acid by the enzyme 5-LO assisted by FLAP. It exerts its actions
by ligating two G proteins-coupled receptors, BLT1 and BLT2. LTB4 chemoattractant function has also been shown for CD4+,
CD8+ and T cells. Another T cells subset are the regulatory T cells (Tregs). Treg are supressive cells that modulate the
immune system response. The aim of our study is to evaluate the LTB4 effect on Treg cells. As we already shown that Tregs
express BLT1 receptor and it is responsive to LTB4 as a chemoattractant factor, our next goal was to evaluate if the LTB4 can act
synergistically with the CCL17, a Treg cells chemoattractant. Also, as we saw in our data that all Tregs from septic mice express
the BLT1 receptor, we sought to analyze if in vitro activated Tregs show the same profile
Splenic cells from C57Bl/6 (B6) were subjected to an EasySep Treg Kit for purification (StemCell, USA), according to the
manufacturers instructions. Purified Treg were placed in the upper well of a Boyden chamber and in the bottom wells 0,1nM
LTB4 and 1nM CCL17 diluted in medium was loaded. As negative control we used RPMI medium only. Migrated cells were
collected after 1h incubation at 37C and counted by hemocytometer. For activation experiments, Tregs were stimulated with
anti-CD3 (5ug/mL), anti-CD28 (2ug/mL), LPS (100ng/mL) and in some experiment with macrophages (3x105). All experiments
were conducted in accordance with the ethical guidelines of the Institutional Animal Care Committee-CEUA in UFRJ, Rio de
Janeiro, RJ, protocol code: DFBCICB028.

Conclusions:
LTB4 did not show a synergistic activity with the CCL17 chemokine in the chemotaxis assay. BLT1 expression was not upregulated on Tregs activated by different stimulus. We have provided data that Tregs express BLT1 receptor and it is responsive
to LTB4. We also showed that LTB4 cant act synergistically with CCL17, and that activated Tregs do not up-regulate BLT1
receptor in vitro. Our next goal is to evaluate the actions of LTB4 in the Treg functions and signaling.
Keywords: chemotaxis, leukotrienes, regulatory T cells
Resumo:18-053
THE ROLE OF LAMININ POLYMER IN SIGNALING OF SPLENIC DENDRITIC CELLS
Ladislau, L. 1; Da-f, A. R. 2; Barja-fidalgo, T. C. 2; Benjamim, C. F. 1

2
DEPTO FARMACOLOGIA E PSICOBIOLOGIA/UERJ, IBRAB/UERJ
1
INSTITUTO DE CINCIAS BIOMDICAS/UFRJ, ICB/UFRJ
Objectives:
Dendritic cells (DCs) are considered professional antigen-presenting cell (APCs) and they are very important during immune
response. Laminin (LN) is a major protein, important and biologically active part of the basal lamina, which plays a role in the
cell differentiation, migration, adhesion as well as phenotype and survival of several cells. For APC function, DCs have to
display foreign antigen by the major histocompatibility complex (MHC), express Toll-like receptors and co-stimulatory
molecules on its surface and release cytokines. We showed that LN increases MHC and co-stimulatory molecules expression on
DCs when they were incubated with and without LPS. However we showed that LN did not induce cytokines release from DCs
when they were incubated with and without LPS. Its known that DCs express 61 integrin, LN receptor, but the effect of LN on
DCss signaling is poorly understood. Our aim is to characterize the effect of LN polymer on signaling of splenic DCs.
This animal study was performed in accordance with instructional guidelines, approved by Ethical committee of animal use,
protocol number: DFBCIB 028. C57BL/6 mice were gently donated by National Institute of Cancer (INCA). The LN (50 mM)
was diluted in sodium acetate pH 4,0 or Tris pH 7,0, both with additional calcium chloride (1 mM), and this LN was kept
overnight in 6-well plates at 37oC and 5% CO2. We used BSA (50 mM) for the protein control. DCs were obtained from spleen
with aseptic techniques and subjected to an EasySep CD11c-PE positive selection (StemCell Technologies), according to the
manufacturers instructions. DCs purity, was determined by flow cytometry of CD11c+ cells and showed >90% after selection.
These cells were cultured in 6-well plates pre-coated with LN or BSA. We also used a second hit of LPS (1g/mL). To analyze
the protein signaling by western blotting, DCs were incubated with stimulus for 30 minutes and 24 hours. We did not observe any
difference at phosphorylation of p38 and ERK in DCs incubated with LN polymer and BSA without LPS in 30 minutes.
However, when DCs were incubated with LN polymer and LPS an increase at ERK phosphorylation was observed, compared
with BSA with LPS group. No difference was observed at p38 phosphorylation. Further, DCs incubated with LN polymer
without LPS for 24 hours presented an increase at p38 and ERK phosphorylation, compared with BSA alone and with LPS
incubation. DCs incubated with LN polymer and LPS presented an increase at p38 and ERK phosphorylation compared with DCs
incubated with LN without LPS. To analyze NFkB activation, EMSA was performed after 1,5 hours of DCs incubation with LN
polymer and BSA with and without LPS. DCs incubated with LN polymer and LPS have more activated NFkB than DCs
incubated with BSA and LPS. In DCs incubation without LPS no difference was observed.
Conclusions:
These results suggest that LN activates phosphorylation of signaling protein and seems to be responsible for long-term
phosphorylation. Also, LN polymer is able to enhance NFkB activation by LPS.
Keywords: DENDRITIC CELLS, LAMININ, SIGNALING, LPS
Financial Support: FAPERJ AND CNPQ
Resumo:18-054
EFFECT OF POLYMER-COATED GOLD NANOPARTICLES ON MACROPHAGE RESPONSE IN VITRO
Albuquerque, A. L. 1; Santos, R. V. 1; Silva, J. P. N. 1; Giacomelli, C. 2; Hickmann, J. M. 1; Santos, C. E. A.

1
; Brito, F. D. A. 1; Martins, M. A. 3; Cordeiro, R. S. B. 3; Barreto, E. O. 1
1
Laboratrio de Biologia Celular, UFAL
2
Departamento de Qumica, UFSM
3
Laboratrio de Inflamao, IOC-Fiocruz
Objectives:
Multiple research groups have tried to apply nanoparticles in a variety of fields such as biological research, medicine and
cosmetics. Among them, the majority were focused on gold nanoparticles as it has been reported to be neither cytotoxic nor to
elicit the activation in macrophage cells. In addition, little is known about the effects of such polymer-coated gold nanoparticles
in biological systems. Therefore, the aim of this study was to evaluate the influence of polymer-coated gold nanoparticle on
macrophages in vitro.
Characterization of the gold nanoparticles (AuNP) with or without polymer-coated (AuNPc) was performed by using UV-VIS
spectroscopy. Macrophages were obtained by intraperitoneal lavage from Swiss mice (18-25g). Peritoneal fluid was cultured for
2 h to allow the macrophage adherence on plastic wells. The supernatants were harvested to remove non-adherent cells.
Peritoneal macrophages were maintaining in RPMI-1640 medium supplemented with Fetal Cow Serum (10%), ciprofloxacin (2
mg/mL) and glutamine (1%). Macrophages were incubated for 12, 24 and 48h with AuNPc or AuNP and phagocytic activity,
morphological analyses, cell viability and reactive oxygen species generation (ROS) were evaluated. The amount of nanoparticles
incubated per well was 100, 101, 102 and 103 ng. Results were expressed as mean SEM and analyzed statistically by the test t
of Student. P values 95%) and uptake of zymosan by macrophages in all doses tested. Moreover, in different points analyzed (12,
24 and 48 h) the nanoparticles did not interfered on cell viability and or phagocytosis. Curiously, AuNP and AuNPc in the high
concentration (103 ng) induced significant cytoplasm vacuolization in the macrophages.
Conclusions:
These results suggest that gold nanoparticle or polymer-coated gold nanoparticle show lower propensity in inducing cell toxicity,
suggesting their bio-compatibility.
Keywords: GOLD NANOPARTICLES , MACROPHAGE , POLYMER-COATED
Resumo:18-055
EXOSOMES AND WASP IMMUNODEFICIENCY
Sunahara, K. K. S.
Medicine SOlna Allergy and Immunology department KI, KI
Objectives:
To verify how changes in the actin machinery can affect exosome production. To characterize and compare exosomes
derived from B cells and bone-marrow derived dendritic cells from WT and WASP-KO mice
Spleen and bone marrow from 6 to 8 weeks BALB-c, WT and WASP-KO mice were processed. B cells were obtained by
negative selection of splenocytes using magnetic beads and stimulated with anti-CD40 antibodies and IL-4 to induce
immunoglobulin class switching. Bone marrow-derived dendritic cells (BMDDCs) were generated using GM-CSF and IL-4, and
on the 6th day of culture, cells were stimulated with LPS for 48 hours to induce DC maturation. WASP-deficient B cells show
higher exosome production when compared to activated bone marrow-derived dendritic cells. Exosomes were phenotyped for
different markers such as tetraspanins (CD9 and CD81), co-stimulatory molecules (CD80 and 86), and the adhesion molecule
CD54. Substantial differences between WT and WASP deficient cells were not observed. However, MHC II expression was
strikingly diminished on WASP deficient activated B cells derived exosomes, while on activated BMDDCs, it was slightly
increased
Conclusions:
WASP deficiency differently affects exosome production and phenotype in B cells and DCs. It would be possible then to
speculate that WASP deficient cells release exosomes that show impaired function, what could compromise the crosstalk among
immune cells
Keywords: exosomes, WASP, immundeficency, auto-immunity
Financial Support: karolinska INstitutet
Resumo:18-056
S100B SECRETION IS STIMULATED BY CYTOKINES IN GLIAL CULTURE AND HIPPOCAMPAL SLICES OF
RATS: LIKELY INVOLVEMENT OF WAY MAPK.
de Souza, D. F. ; Hansen, F. ; Gonalves, C. A.

Departamento de Bioqumica/ICBS, UFRGS
Objectives:
S100B is a cytokine produced and secreted by astrocytes, has trophic or toxic depending on concentration, is involved in
neuroinflammatory processes, present in neurodegenaratives diseases such as Alzheimer's disease, schizophrenia, among others.
However, its mechanism of secretion has not been elucidated. In this study, we investigated S100B secretion in C6 glioma cells
and acute hippocampal slices exposed to IL-1, IL-6, IL-8 and TNF-.
We used C6 glioma cells and hippocampal slices from rats exposed or not to cytokines and / or specific inhibitors of MAPK
pathways. Quantification of S100B was performed by ELISA. We used t test or one-way ANOVA assuming p
Conclusions:
Both PD98059, an inhibitor of ERK, and SB203580, p38 kinase inhibitor, were able to prevent the increase in S100B secretion
stimulated by cytokines, suggesting that this effect, observed in two different preparations in vitro, is mediated by signaling
pathway MAPK- ERK. These results contribute to the inference that S100B secretion induced by cytokines, is a component of
the neuroinflammatory response, which would support the involvement of S100B in the genesis of neurodegenerative diseases.
Keywords: S100B, Cytokines, MAPK, Neurodegenerative disease
Financial Support: CNPq, FINEP, CAPES
Resumo:18-057
TLRS ENHANCE FCR-MEDIATED PHAGOCYTOSIS VIA 5-LO PRODUCTS IN MACROPHAGES
Pinheiro, C. S. 1; Teixeira, A. P. M. 1; Benjamim, C. F. 2; Peters-golden, M. 4; Bozza, M. T. 3; Canetti, C. 1

1
Instituto de Biofisica Carlos Chagas Filho, IBCCF
2
Instituto de Ciencias Biologicas, ICB
3
Instituto de Microbiologia Professor Paulo Goes, IMPPG
4
University of Michigan, UM
Objectives:
Toll-like receptors (TLRs) are pattern recognition receptors involved in microorganisms recognition, which are engulfed by a
mechanism known as phagocytosis. Leukotrienes (LTs), lipid mediators produced from 5-lipoxygenase (5-LO) pathway, are able
to increase phagocytosis in FcgR dependent process. They are produced from arachidonic acid metabolism in a two-step process
to yield the epoxide intermediate, LTA4 which is then converted into either LTB4 by LTA4 hydrolase or conjugated with
glutathione by LTC4 synthase to form the cysteinyl LTs (cys LTs). The aim of this study was to evaluate the participation of
TLRs ligands on FcgR-mediated phagocytosis and the role of 5-LO products.
Methods: Rat alveolar macrophages (AMs) or mouse (SV129 or 5-LO-/-) peritoneal macrophages (PMs), and bone-marrow
derived macrophages (BMDM) were harvested, cultured, and treated with TLR ligands and then challenged with IgG opsonised
sheep red blood cells (IgG-sRBCs). Zileuton, a 5-LO inhibitor, was incubated before TLR ligands addition. After 90 min, the
phagocytic targets were removed and phagocytosis evaluated through a colorimetric assay. Lipid mediators (LTB4 and cys LTs)
were measured by EIA. Western blotting, was used to detect activation and expression of mitogen-activated protein kinase
pathway. Results: Except for CpG, all TLR ligands tested (i.e ligands for TLR2, TLR3, and TLR4) were able to amplify
phagocytosis of IgG-sRBC in AM and BMDM. The phenomenon occurred in a concentration-dependent manner. Futhermore,
phagocytosis amplification induced by TLR3 ligand was a time-dependent event, fact not observed for TLR2 and TLR4 ligands.
AMs treatment with zileuton also impaired TLR4 ligand-induced phagocytosis amplification, suggesting the participation of 5LO products in this phenomenon. To confirm LTs involvement in phagocytosis mediated by FcgR, PMs from 5-LO-/- mice did
not presented any difference on phagocytosis when stimulated with TLR2, 3, and 4 ligands. LTB4 production induced by IgG
engagement was amplified by TLR2 and TLR4 ligands, but not by TLR3. Similar results were obtained for cys-LTs, whose
production was increased by TLR2, TLR3, and TLR4. ERK1/2 and p38, mitogen-activated protein kinase (MAKs) pathways
were phosphorilated under TLRs agonists. This phenomenon was exacerbated under IgG-sRBC treatment.
Conclusions:
Our data suggest that TLR ligands amplify phagocytosis in a time and concentration dependent manner. Furthermore, 5-LO
products pathway seems to be responsible for this phagocytosis amplification. ERK1/2 and p38 presented a greater activation
when cells were incubated with TLRs agonists and challenged with IgG-sRBC, suggesting a possible interaction between FcgR
and TLR signaling pathway.
Keywords: phagocytosis, TLRs, Leukotrienes
Financial Support: FAPERJ, CNPq, and FUJB
Resumo:20-001
INHIBITION OF RT HIV-1 BY DITERPENES ISOLATED FROM BRAZILIAN BROWN SEAWEEDS: A
STRUCTURE-ACTIVITY RELATIONSHIP STUDY
Miceli, L. A. 1; Souza, A. M. T. 1; Teixeira, V. L. 1; Paixo, I. N. P. 1; Rodrigues, C. R. 2; Castro, H. C. 1

1
Departamento de Biologia Geral (GBM), UFF
2
Faculdade de Farmcia da UFRJ, UFRJ
Objectives:
The marine organisms are source of numerous molecules presenting distinct and important roles such as chemical defense against
herbivores. Diterpenes isolated from Brazilian brown seaweeds of the genus Dictyota have a promising and potential activity as
antiviral against HIV. Interestingly, the diterpene 8,10,18-trihydroxy-2,6-dolabelladiene (THD), (6R)-6-hydroxydichotom-3,14diene-1,17-dial (HDD) and (6R)-6-acetoxydichotom-3,14-diene-1,17-dial (ADD) showed a significant antiviral activity against
reverse transcriptase (RT-HIV-1) of HIV-1 (IC50=16.5uM, 10M and 35M, respectively) and low cytotoxicity (CC50 >200
M). The aim of the present work is to study the Structure-Activity relationship (SAR) of these diterpenes as anti RT-HIV-1
agents to identify important parameters related to this biological profile, using a molecular modeling approach.
Stereoelectronic parameters from diterpene compounds were obtained from Single-Point HF/6-31G* calculations using
SPARTAN08 program. Among all ten parameters calculated, the overall analysis revealed a feasible relationship of the anti RTHIV-1 activity with GAP (ELUMO-EHOMO), which represents the molecule reactivity. Higher values of GAP led to higher
antiviral profile in contrast to the polarizability and dipole moment values where higher values negatively affected it. Since ADD
presents the highest molecular weight (366 Da) and the lowest antiviral activity profile, probably the size of the molecule may
also influence the derivative interaction with the RT-HIV-1 active site. In this work we calculated the theoretical toxicology of
these compounds. Interestingly, the theoretical toxicity results were analogous to the previous experimental data where THD
showed a lower cytotoxicity (CC50=500 M) than HDD and ADD (CC50 = 200 M) and nevirapine (CC50>100 M). Finally,
we calculated the theoretical lipophilicity (cLogP) and tested the fulfillment of the Lipinski rule-of-five for all compounds
using Osiris Property Explorer server. Our results suggested that the compounds may have a good oral availability as they
fulfilled the Lipinski rule-of-five and presented higher cLogP values.
Conclusions:
According to our SAR study, dipole moment, polarizability, GAP and molecular weight should be carefully considered when
designing new anti RT-HIV-1 agents since they apparently influence the antiviral profile. Overall THD may be pointed as a
promising lead molecule for further exploration by using a molecular docking approach.
Keywords: HIV-1, Reverse Transcriptase, Diterpenes, Molecular Modeling, Antiviral
Financial Support: UFF-PROPPI, CNPq, FAPERJ
Resumo:20-002
CRYPTOCOCCUS NEOFORMANS CYP51 ENZYME: HOMOLOGY MODELING AND INTERACTION WITH
AZOLE ANTIFUNGALS
Carvalho, K. L. 1; Abreu, P. A. 3; Castro, H. C. 1; Rodrigues, C. R. 2

1
Depto de Biologia Celular e Molecular / Inst. de Biologia , UFF
2
Faculdade de Farmcia, UFRJ
3
Faculdade de Farmcia, UFRJ / Campus Maca
Objectives:
Cryptococcus neoformans is an opportunistic fungus involved in central nervous system infections, which are fatal if untreated. It
is also extremely important in affecting immunocompromised individuals. Fluconazole and other azoles antifungal are one of the
main treatments that inhibit lanosterol 14-demethylase, a CYP51 family protein. In fungi, CYP51 participates in ergosterol
biosynthesis essential for fungal viability. Currently, the emergence of resistant strains and the cross-binding of known azoles
with human CYP51 demand the understanding of CYP51 structure and the search for more selective compounds. Since the
experimentally 3D-structure of the CYP51 of C. neoformans is not described in the literature, the aim of the present study is to
construct an homology model of this enzyme and dock it with known azoles to evaluate their interactions. We also docked these
inhibitors into the human CYP51 crystal structure for further comparison.
The multiple sequence alignment of the C. neoformans CYP51 and other CYP51 primary sequence using CLUSTALW revealed
low overall identity percentage ranging from 25 to 46%. Nevertheless, the secondary structure was conserved for all sequences
that also presented a similar folding prediction or crystal structure. The Swiss-model program was used to construct the C.
neoformans CYP51 homology model using the human crystal structure as template due to its higher degree of similarity (36%)
compared with other 3D-structures currently available (i.e. Mycobacterium tuberculosis CYP51 = 28%). The model was refined
using molecular mechanics and the reliability was assessed by Ramachandran plots and Profile-3D that confirmed the feasibility
of the structure. The C. neoformans model was similar to the template with 15 -helix and 5 -sheet. However, the electrostatic
potential map analysis revealed a more positive profile for the human CYP51 compared with the C. neoformans CYP51
homology model. The theoretical studies with ketoconazole and fluconazole, constructed and minimized using the program
Spartan 8.0, and docked into human and Cryptococcus enzymes using the Autodock program, showed the same binding mode
involving the iron of the CYP51 Heme group and the cysteine 490. Interestingly, the study pointed to a higher binding energy of
the ketoconazole and fluconazole to the C. neoformans CYP51 compared with the human enzyme. In addition, the comparison of
each enzyme active site (within 6 from the inhibitors) revealed different residues interacting and/or near to the compounds
including Y77F, L100M, R103K, I105F, A130V, V144A, I159L, P242H, F245W, M246L, A313G, M316L, S388I, I389M,
Y390M.
Conclusions:
Compared with the human enzyme, the CYP51 from C. neoformans presents differences in the electrostatic charge distribution
and in residues involved in the inhibitor interaction. Our study pointed to new regions to be further explored on designing new
more effective antifungals.
Keywords: Homology modeling, Cryptococcus neoformans, 14-lanosterol demethylase
Financial Support: FAPERJ, CNPq, CAPES, UFF-PROPPi
Resumo:20-003
MOLECULAR DOCKING APPROACH FOR STUDYING PIPERAZINE DERIVATIVES: A NEW SERIES OF NMDA
RECEPTOR ANTAGONISTS
Santana, M. V. D. S. 1; Abreu, P. A. 2; Castro, H. C. 1

1
Depto. de Biologia Celular e Molecular/Instituto de Biologia, UFF
2
Faculdade de Farmcia/Universidade Federal do Rio de Janeiro, UFRJ
Objectives:
The N-methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate-gated channels that are essential to synaptic plasticity,
neuronal development, memory and learning. The NMDARs are composed of different subunits as NR1, NR2 (A-D) and NR3 (A
and B). The excessive stimulation of the NMDAR is associated with some pathological conditions as Parkinson, Huntington and
Alzheimer diseases. Because of its role in neuronal death, NMDA receptor antagonists is the focus of intense research, especially
involving selective NR2B subunit antagonists. As NR2B is the only subunit absent in the cerebellum, the block of this subunit
results in less adverse effects as they may not interfere with the motor function. In this context, the aim of this work is to analyze
the interactions and the binding mode of 17 piperazine-2,3-dicarboxylic acid derivatives (13, 17, 23, 16a-n) within NR2B subunit
of NMDA receptor.
For docking the piperazine derivatives we used Autodock 4.2 software that calculates ligand interactions within the receptor
active site. On that purpose three steps were performed including preparation of the structure files, pre-calculation of the possible
interacting atoms in the ligand (AutoGrid) and the docking of the molecules based on the information of the grid. The results of
docking analysis showed a region within the receptor active site characterized by a hydrophobic pocked compose of the residues
Tyr212, Asp213, Thr241, Thr172, Ser171 and Glu13, which interact with the hydrophobic core of the ligands. Compounds
bearing bulky hydrophobic groups (16e, 16g, 16h and 16n) were the most potent compounds. They interact with the same
residues in the active site with a similar binding mode. The most active ligands presented hydrogen bonds with Lys85 and His86,
as most of the other derivatives. 16h derivative showed a different interaction pattern and a slightly different conformation
compared with 16e, 16g and 16n, that did not affect the biological activity. Interestingly, compound 16g, with the best Ki value,
also presented the lower binding energy. Analogously, 16a, with the worst Ki value, also presented the higher binding energy,
which may be related to its completely different conformation from the most active molecules.
Conclusions:
Our results suggest that interactions of bulky hydrophobic groups of the ligands with the NMDA receptor are determinant for the
biological activity as the most potent molecules of this series presented a phenanthrene ring as substituent that directly interact
with the receptor. The hydrogen bonds detected within the active site were almost the same for the compounds pointing to the
importance of van der Walls interactions of the hydrophobic groups of 16e, 16g, 16h and 16n, the best antagonists present in the
study. This study may help in the rational design of more potent and selective drugs for neurodegenerative diseases.
Keywords: Autodock, Piperazine derivatives, Molecular docking, NMDA, NR2B
Financial Support: CNPq, FAPERJ, UFF-PROPPi, CAPES
Resumo:20-004
A SYSTEM BIOLOGY APPROACH ON THE EVOLUTION OF CHEMOSENSORY RECEPTORS IN MICE, RATS
AND HUMANS
Albanus, R. D'o. ; Dalmolin, R. J. S. ; Moreira, J. C. F.

Instituto de Bioqumica, UFRGS
Objectives:
The aim of the present study is the use of System Biology analysis tools in the assessment of evolutionary parameters of the
chemosensory networks, namely taste, olfactory and vomeronasal reception, on Mus musculus, Homo sapiens and Rattus
norvegicus.
Using data gathered from The Gene Ontology Project (GO), we were able to amass the genes associated with each type of
sensory perception. The GO IDs used for this work were 0004984 and 0007608, 0050909 and 0008527, 0016503 and 0019236,
for olfactory, gustatory and vomeronasal ontology groups, respectively. We sorted these genes in groups by receptor modality,
with one further division of the Taste Group into the Taste-1 and Taste-2 receptors types groups. We acquired the network
parameters of the proteins translated from these genes groups using data from STRING Database. We used as criterions for our
analysis the connectivity and clusterization indexes (i.e. k(i) and c(i), respectively). Also used in our analysis was the
Evolutionary Plasticity Index (EPI) of proteins orthologous group, an indicator that is able to infer the evolutionary constraints
acting on its evolution. We subsequently divided the whole dataset into two more groups: those of proteins directly involved in
the chemical binding of the stimuli, named Receptor Group (RG); and those involved into signal transduction and further
chemical pathways, named Accessory Group (AG). We compared the mean EPIs of all the RGs and AGs using One-Way
ANOVA and Tukey test and found that the AGs means were significantly lower than the RGs (p<0,05).
Conclusions:
Our findings suggest that the evolutionary plasticity in the RGs is higher than that on the AGs. This means that the selective
pressures among the chemoreceptors are weaker than that on the transduction machinery, resulting in a greater diversity for these
receptors. The peaks of clusterization and connectivity for the Olfactory RGs are probably due to the fact that the expression of
one olfactory receptor apparently inhibits the others, in an one-neuron-one-receptor model. Put together, these results support the
Birth-and-Death Evolution model proposed for chemosensory receptors and others genetic families with notably high degree of
variability among its members. This work also demonstrate the value of System Biology as an efficient and cost-compatible
analysis tool to study complex interactions between proteins networks, being able to infer evolutionary relationships based on the
mathematical (and therefore, comparable) data generated by this kind of approach.
Keywords: Chemoreception, Evolution, System Biology
Financial Support: This work was funded by CNPQ and Capes.
Resumo:20-005
ANALYSIS OF THEORETICAL MODELS FOR ATP PRODUCTION: CONFLICTS AND CORRELATIONS
AMONG THE MODELS AND WITH EXPERIMENTAL DATA
Siqueira, K. M. ; Saa, A.
Dept. of Applied Mathematics / IMECC, UNICAMP
Objectives:
In 1997, Magnus and Keizer (Am. J. Physiol. 273, C717, 1997) first proposed a theoretical dynamic model for ATP production in
the mitochondria (MK model) based on the biophysical properties of the enzymes and transporters involved in the process. This
model is very complete and takes into account most of the processes thought to be important for mitochondrial oxidative
phosphorylation. However, some biological properties are difficult do derive from the MK model due to its complexity. A few
years later, a simplification of this model was proposed by Bertram et al. (J. Theor. Biol. 243; 575, 2006), and, although this
model has been used in several works since its publication, is hadnt yet been fully tested as to its correlation with the MK model
and with current biological data. Therefore, the purpose of this work was to evaluate the validity of the modifications proposed by
Bertram et al., by comparing the predictions of this model with the MK model and with current biological data.
MK model and Bertrams model are composed of a group of equations that describes the dynamics of the citric acid cycle, the
proton pump as well as ATP and calcium transporters at the inner mitochondrial membrane. Weve studied each of Bertrams
equations in comparison to the MK model. By graphical analysis, the general behavior of Bertrams equations was compared
both with the MK model and with current biological data from the literature. For three equations, we found that the Bertram
Model had a significantly different comportment (even qualitatively) from MK model. These equations described the rate of
production of NADH by the citric acid cycle, of the adenine nucleotide translocators activity and of the calcium uniporter
transporter. For these three rates, Bertrams model didnt agree with the biological data, while the MK model described it
accurately. Here, we proposed some modifications of the MK model, that provided simpler equations and still presented a good
correlation with MKs. The other equations from Bertrams model agreed with the MK model as well as with the biological data.
Conclusions:
We found that three equations from Bertrams model didnt agree with biological data. Apart from that, both models (MK and
Bertams) presented a good correlation with biological data, suggesting, therefore, that they can be powerful tools in the study of
ATP production dynamics.
Keywords: ATP production, Biophysics, Mathematical Model, Mitochondria
Resumo:20-006
ALTERNATIVE METHODS OF TEACHING PHARMACOLOGY: FULFILLING AROUCAS LAW.
Chaves, H. V. ; Saraiva, T. V. ; Cruz, M. P. ; Vieira, A. M. ; Silva, W. C. B. ; Oliveira, I. N. S. ; Camelo, J.

R. R. ; Val, D. R. ; Silva, A. A. R. ; Bezerra, M. M.
Objectives:
The practical activities in pharmacology teaching had to suit the new legislation enacted in 2008 as the Aroucas Law, which
states that "whenever possible, teaching practices should be photographed, filmed or recorded, allowing its reproduction for
future practices, avoiding the unnecessary repetition of procedures teaching with animals." In this regard, the aim of this study
was to develop software that will be used for teaching practical classes in pharmacology in accordance with Aroucas Law,
stimulating the interdisciplinarity between the faculties of Medicine and Computer Engineering from the UFC - Campus of
Sobral.
We observed the classroom practice of routes of administration of drugs held in the Principles of Pharmacology module at
Faculty of Medicine of Sobral. Chloral Hydrate (10mg/kg), a sedative-hypnotic agent, was injected in six female Wistar rats (200
220g) via oral, subcutaneous and intramuscular, and the time to onset of drug effect was recorded. For inhalation and
intravenous routes a swab of cotton wet with ether and Evans blue (1%), has been used, respectively. To record the videos we put
the animals in a glass box (40x40x30cm) and used a high definition camera and fluorescent lamps tilted 45 degrees from the
sidelines. There is a menu at the initial screen of the software containing the presentation, objectives, methodology, videos and
results. There is also an area with graphics and issues related to practice with the purpose of exercising the content worked.
Conclusions:
The software is an alternative method of teaching pharmacology. Moreover, its development has encouraged the interdisciplinary
between exact and life sciences.
Keywords: Alternative Methods , Arouca's Law, Computer Engineering , Software , Teaching Pharmacology
Financial Support: UFC
Resumo:20-007
DIPHENYL DISELENIDE INHIBITS MAMMALIAN AND DROSOPHILA MELANOGASTER BUT NOT PLANT
&DELTA;-AMINOLEVULINIC ACID DEHYDRATASE ENZYME: A MOLECULAR MODELING APPROACH.
Bueno, D. C. ; Nogara, P. A. ; Saraiva, R. A. ; Rocha, J. B. T.

Depto de qumica - Universidade Federal de Santa Maria, UFSM
Objectives:
In the last years, diphenyl diselenide [(PhSe)2] has been suggested as a potential therapeutic agent, with neuroprotective,
analgesic and antioxidant effects both in vivo and in vitro. Delta-aminolevulinic acid dehydratase (-ALAD) is a metalloprotein
that catalyzes porphobilinogen (PBG) formation. Since -ALAD is important for heme synthesis, its inhibition can be toxic.
Previous studies have demonstrated that (PhSe)2 inhibits human, murines and fly (Drosophila melanogaster) -ALAD by
oxidation of its sulfhydryl residues. On the other hand, cucumber (Cucumis sativus) -ALAD is not inhibited by (PhSe)2. So, we
compared -ALAD structures from mammals, fly and cucumber and performed molecular modeling studies using protein-ligand
docking tools, in order to get a better understanding of in vitro effects observed.
FASTA sequences from Homo sapiens -ALAD (Hs-ALAD), Mus musculus -ALAD (Mm-ALAD), D. melanogaster ALAD (Dm-ALAD) and C. sativus -ALAD (Cs-ALAD) were obtained from BLAST. The crystal structure from Hs-ALAD
was obtained from Protein Data Bank (PDB ID: 1E51) and Dm-ALAD was modeled using Swiss-MODEL server. Virtual
docking was performed by AutoDock Vina 1.1 program. Avogadro program was used to construct the ligands, and they were
optimized using UFF method. The grid box was centered in the following coordinates: x = 37.1, y = 74.9 and z = 58.01, and box
size was defined as 20 x 20 x 20 (spacing = 1 ). Docking simulations with Hs-ALAD showed that the aromatic rings of
(PhSe)2 make - interactions with Phe79, - interactions with Phe208, and cation- interactions with Lys199 and Arg209,
allowing an approximation between Se atoms and thiol group of Cys124, with distances ranging between 3.3 and 3.4 .
Docking simulations with Dm-ALAD showed that the aromatic rings of (PhSe)2 make - interactions with Phe204 and cation-
interactions with Lys195 and Arg205 also allowing an approximation between Se atoms and thiol group of Cys124, with
distances ranging between 3 and 4 . In spite of some modifications between Dm-ALAD, Mm-ALAD and Hs-ALAD
active site are observed, the multiple sequence alignment show that the residues related to the activity are conserved. However,
when comparing Hs-ALAD with Cs-ALAD, tyrosyl, alanyl and aspartyl residues in place of cysteyl residues are observed.
Conclusions:
The results obtained are consistent with experimental data. The approximation between Se atom from (PhSe) 2 and Cys124 allows
the oxidation of thiol groups, inhibiting porphobilinogen synthesis. According to the analysis of multiple sequence alignments,
the substitution of cysteyl residues in Cs-ALAD could explain why (PhSe)2 does not inhibit it, suggesting the importance of
cysteyl residues in Hs-ALAD and Dm-ALAD inhibition by (PhSe)2.
Keywords: diphenyl diselenide, molecular modeling, phorphobilinogen synthase, thiol oxidation
Resumo:20-008
MOLECULAR VIRTUAL DOCKING AND IN VITRO STUDIES OF SYNTHETIC ALKALOIDS WITH
QUATERNARY AMMONIUM AS ACETYLCHOLINESTERASE INHIBITORS
Saraiva, R. A. 1; Nogara, P. A. 1; Bueno, D. C. 1; Pereira, R. P. 1; Appel, A. S. 1; Paixo, M. W. 2; Rocha, J.

B. T. 1
1
Departamento de Qumica / Bioqumica Toxicolgica, UFSM
2
Departamento de Qumica, UFSCar
Objectives:
Alzheimer's disease (AD) is a neurodegenerative disorder associated with loss of brain neurons and affects a large number of
people around the world. The use of acetylcholinesterase (AChE) inhibitors has been considered one of the strategies to treat AD
since these compounds increase the concentration of acetylcholine (ACh) in the brain. Among the AChE inhibitors, some natural
and synthetic alkaloids with quaternary ammonium (i.e., edrophonium) are largely employed. Despite their benefits, these
compounds present some limitations. Thus, further researches into the development of new AChE inhibitors with less adverse
effects are need. In line with this, we aimed to evaluate the AChE inhibitory activity from eight new compounds with quaternary
ammonium derivatives from the alkaloid cinchonidine (compounds A to H) as well as to understand the interactions involved in
their AChE inhibition by molecular docking in silico.
IC50 from compounds was estimated by AChE activity by Ellman's method in vitro (Biochem Pharm 7: 88, 1961). The
compounds (at concentrations ranging from 0.1 to 100 mol/L) or vehicle 2% DMSO were pre-incubated for 20 min with AChE
prior to 2-min incubation with acetylthiocholine substrate at 25 C. Molecular docking studies were carried out using AutoDock
Vina 1.1 program. The crystal structure of AChE was obtained from Protein Data Bank (PDB id: 2XUF). Ligands were drawn in
Avogadro 0.9 program and their energies were minimized using MMFF94 force field. The active center gorge was centered at
coordinates x = 32.81, y = 24.17, z = 10.59. The analyses of results were performed using PyMOL and Accelrys Discovery
Studio Visualizer 2.5 programs. The pose with lowest binding free energy was accepted as the most probable interaction model
for each docking procedure. According to in vitro assay, compounds B (0.80 mol/L), F (2.80 mol/L), D (7.90 mol/L) and C
(18.10 mol/L) showed the lowest IC50 when compared to other compounds (IC50 > 40 mol/L). Taking into account the
compounds B, C, D and F, together, molecular docking analysis demonstrated an approximation of quaternary ammonium close
to residues from anionic subsite (Trp86, compound F) or peripheral anionic subsite (Trp286, compounds B, C and D) of AChE by
cation- stacking interactions. - stacking interactions between aromatic groups from these compounds and aromatic residues
(Trp286, Tyr 72 and Tyr341) was also observed. The compounds D and F make H-bond involving the residue Asp74.
Conclusions:
The proposed molecular models suggest an important role of peripheral aromatic residues (Trp, Phe and Tyr) from AChE active
site gorge in the interaction involving both the aromatic moieties and the quaternary ammonium from compounds that could lead
to blocking the entrance of ACh in the enzyme. These data also point that the compounds B, C, D and F could be used as drugs in
the treatment of AD and more pre-clinical studies should be conducted in order to understand their in vivo effects.
Keywords: acetylcholinesterase, virtual docking, Alzheimer's disease, quaternary ammonium
Financial Support: CNPq, FAPERGS
Resumo:20-009
CIS-REGULATORY CONTROL OF PATTERN FORMATION IN DROSOPHILA EMBRYONIC DEVELOPMENT.
Lopes, F. J. P. 1; Cardoso, M. A. 2; Vieira, F. D. M. C. 3; Holloway, D. M. 4; Spirov, A. V. 6; Bisch, P. M. 2

1
Instituto de Biofisica Carlos Chagas Filho, UFRJ Xerm
2
3
Instituto de Qumica, UnB
4
Mathematics Department, BCIT
6
Center for Developmental Genetics, SUNYSB
Objectives:
During Drosophila embryonic development the concentration gradients of maternal factors, like the Bicoid protein, establish
positional information along the embryo. A cascade of developmental genes reads out this positional information by exhibiting
different activation levels according to their position along these gradients. One of the first genes of this cascade, the gap gene
hunchback (hb) has strong anterior expression and a sharp on-to-off gradient which is essential for regulating pattern in the midembryo. It has been shown that Bcd binds to hb promoter cooperatively and that hb activates its own regulation. The role of Bcd
cooperative binding for Hb pattern positioning has been demonstrated already, but the mechanism that allows the shallow Bicoid
gradient to regulate the sharp Hb border remains an unsolved problem.
We used immunohistochemistry to quantify the dynamics of hb gene expression in flies that were wild-type, were mutant for hb
self-regulation or Bcd binding, or contained an artificial promoter construct consisting of six Bcd and two Hb sites. In addition to
these experiments, we developed a reactiondiffusion model of hb transcription, with Bcd cooperative binding and hb selfregulation. We found that bistability stemming from hb self-regulation produces the sharp Hb border; the loss of sharpness for the
hb14F self-regulating mutant and the shallow pattern of a Bcd-dependent lacZ artificial construct support this result. In addition,
our results indicate that Bcd cooperative binding determines the position of Hb pattern by determining the position where
bistability occurs; our Bcd cooperative mutant data support this conclusion PLoS Comput Biol 4(9), 2008.
Conclusions:
The ability to produce sharp borders is a central step for the expression of developmental genes in Drosophila and other
organisms, and we show that spatial bistability can play a key role in this process.
Keywords: Development, Pattern, Drosophila, Embryo, Morphogen
Financial Support: CNPq, NIH (2-ROIRR07801) , NSF/NIGMS (1-R01-GM072022) and FAPERJ
Resumo:20-010
COMPUTATIONAL MODELING OF NEW DENTATE GYRUS GRANULE CELLS BORN AFTER STATUS
EPILEPTICUS INDUCED BY PILOCARPINE
Tejada, J. 1; Arisi, G. M. 1; Cairasco, N. G. 1; Roque, A. C. 1

1
Depto. de Fsica - FFCLRP, USP
2
Depto. de Fisiologia - FMRP, USP
Objectives:
Status Epilepticus (SE) provokes changes in the morphology of dentate gyrus (DG) granule cells (GCs) (Epilepsia, 50:2638,
2009; J. Neurosci. 17:3727, 1997; Brain Res 1165:126, 2007.). The new DG GCs have shorter dendrites, narrower arborizations
with greater number of branches (Brain Res 1165:126, 2007) and, in some cases, a basal dendrite that is projected towards DG
hilus (J. Neurosci 27:9400, 2007). The aim of this work is to evaluate how these morphological differences affect the
electrophysiological properties of DG GCs, using as tool computer simulations of biophysically detailed multicompartmental
models.
We worked with a sample of 40 tridimensional reconstructions of DG GCs including 20 from animals treated with pilocarpine
(PILO) and 20 from control rats (Brain Res 1165:126, 2007). Whole dendritic arborizations of each cell were reconstructed using
Neurolucida (Microbrightfield), available on the internet site of the NeuroMorpho project (http://neuromorpho.org/).
Biophysical information came from models of DG GCs(J. Physiol. (Lond.) 538: 227, 2002) avaliable at NeuronDB
(http://senselab.med.yale.edu/neurondb/). The cell models were constructed in NEURON (Neural Comput 9:1179, 1997). The
ionic channels and their maximum conductance distributions were the same for both control and PILO cell models, and the
objective for this was to evaluate the effects of differences in morphology on the electrophysiological behavior. Modeled cells
were submitted to voltage clamp simulations using 14 different current values from 0.005 nA to 0.075 nA. Currents were injected
in sequence with intervals of 500 ms. A 500 ms delay was used for the first current injection. Currents were injected using
different numbers of electrodes connected randomly to one up to five dendrites. The electrodes were placed in three different
areas of the dendritic tree, which correspond to areas into which DG molecular layer is divided: inner (IML), middle (MML) and
outer (OML) molecular layer (J. Neurosci. 31:105, 2011). Membrane voltages were recorded to generate spike counts and
interspike intervals. Simulation results show that PILO cell models are less excitable, presenting smaller number of spikes and
longer interspike intervals, mainly when stimulated at IML.
Conclusions:
Dendrites of DG new cell models exhibited lower response precisely in the area in which mossy fiber sprouting after SE
generates reverberant circuits, which stimulate DG cells. The smaller IML excitability of PILO cell models, if confirmed
experimentally, can be interpreted as a protective mechanism that allows cells to cope with increased stimulation resulting from
mossy fiber sprouting after SE.
Keywords: Computational neuroscience, Dentate gyrus, Detailed single-neuron modeling, Morphology-function relationship,
Mossy fiber sprouting
Financial Support: FAPESP, FAPESP-CInAPCe, CNPq, PROEX-CAPES, FAEPA
Resumo:20-011
THE ROLE OF MICRORNAS IN THE CONTROL OF APOPTOSIS IN NEURODEGENERATION AN IN SILICO
APPROACH
Sousa, . ; Kihara, A. H.
Objectives:
Neurodegenerative diseases are some of the most common disorders that affect people in every age. Many of these disorders do
not have an effective therapy, and are extremely debilitating for the patients. In the last decade, the study of short RNAs increases
exponentially and have provide important information about pathogenesis and prognostic of several diseases. Nowadays, it is
well defined a new platform for drug development, based on short RNAs, especially microRNAs (miRNAs). These are noncoding oligonucleotides whose activity regulates negatively the translation of target mRNAs, by pairing a complementary
sequence at 3UTR. Each miRNA has potential to regulate many genes, and its function may be different depending on the tissue
and pathophysiological process. Thus, miRNAs analyses are complex because involves gene networks that govern several
activities at the same time. Bioinformatics provides many tools for these analyses, and are essentials for the success of miRNA
research. The aim of this study was to find potential targets for eighteen selected miRNAs through bioinformatics tools and infer
their putative role in neurodegenerative process.
Eighteen miRNAs (miR-7-1, miR-9-1, miR-21, miR-25, miR-96, miR-103, miR-107, miR-124-1, miR-146a, miR-150, miR-191,
miR-204, miR-210, miR-222, miR-223, miR-300, miR-335, and miR-494) were selected by previous described association with
neurodegeneration. Subsequently, we used miRGator (available at http://genome.ewha.ac.kr/miRGator/miRGator.html) and PITA
(available at http://genie.weizmann.ac.il/pubs/mir07/mir07_dyn_data.html) algorithms for prediction of potential targets for these
miRNAs. From this list of target genes, we selected genes expressed at neural tissue using the TiGER algorithm (available at
http://bioinfo.wilmer.jhu.edu/tiger/). The putative processes which these genes are involved were analyzed at Gene Ontology
(available at http://www.geneontology.org/). From this method, we were able to obtain a list of 64 genes involved in apoptosis
which are possible targets of the selected miRNAs. Since criterions for miRNA-mRNA interaction are still in discussion, the
number of target genes ranges from approximately 40 to 60, according to the target prediction method. This data, when analyzed
in parallel to published data, elucidate some cellular processes that, when deregulated, deflagrate apoptosis in neurodegeneration.
Conclusions:
Our in silico analysis permitted the visualization of gene interactive networks and provided data that may trigger studies using
experimental approaches. In doing so, it will be possible to find new ways to intervene at the molecular level to prevent damage
caused by neurodegeneration.
Keywords: microRNA, Apoptosis, Neurodegeneration, Retina, Bioinformatics
Financial Support: FAPESP, CNPq and UFABC
Resumo:20-012
ROLE OF ACTIVE DENDRITES IN SHAPING THE RESPONSE FUNCTION OF RETINAL GANGLION CELLS
Publio, R. ; Carlos, A.
Departamento de Fisica e Matematica/FFCLRP, USP
Objectives:
A central problem in contemporary neuroscience is to understand the functional role of voltage-dependent conductances in
neuronal dendritic trees (Annu Rev Neurosci 28:503, 2005). A recent hypothesis for the functional role of active dendrites is that
they enhance the dynamic range of neurons (PLoS Comput Biol 5:e1000402, 2009). The objective of this work is to investigate
this hypothesis using computational models of retinal ganglion cells.
We used a data set of 30 morphologically reconstructed ganglion cell models (J Neurophysiol 81:1685, 1999). The models
received four voltage-dependent (Na, K, Ca, KA), one calcium-dependent (KCa) and a leakage conductance, which were
distributed over their dendrites according to realistic patterns (J Neurophysiol 103:1357, 2010). The number and distribution of
ribbon synaptic locations in a dendritic tree were determined as in (J Neurophysiol 81:1685, 1999). Dendritic sites were
stimulated in various distributions with total excitatory currents in the range of 15-320 pA (J Neurophysiol 81:1685, 1999). We
fitted the experimental EPSP waveform with a double-exponential function. By varying the stimulus current we constructed
input-output response curves that related the firing frequency of the cell with the stimulus current (F-I curve). The dynamic range
was determined from the F-I curve as in (PLoS ONE 4:e6970, 2009). Our results show that the dynamic range depends on the
total dendritic area and the number of branches in the dendritic tree.
Conclusions:
Our results show a strong nonlinear effect of the active dendrites on the F-I curve of retinal ganglion cells. The dynamic range of
our model neurons is affected by the total dendritic area and the number of branches in the dendritic tree.
Keywords: Retina, Ganglion cell, Dynamic Range, Dendrite, Branch
Financial Support: FAPESP, CNPq, INCEMAQ
Resumo:21-106
NEURONAL NITRIC OXIDE SYNTHASE (NNOS) AND CONSOLIDATION OF CONTEXTUAL FEAR
CONDITIONING: EFFECTS OF 7-NITROINDAZOLE, A SELECTIVE NNOS INHIBITOR.
Faria, L. O. M. 1; Canova, F. 1; Vieira, A. S. 1; Teixeira, S. A. 2; Muscar, M. N. 2; Ferrari, E. A. D. M. 1

1
Anatomia, Biol. Celular, Fisiologia e Biofsica - IB/UNICAMP, UNICAMP
2
Departamento de Farmacologia - ICB/USP , USP
Objectives:
This study examined the effects of the intracerebroventricular administration of 7-nitroindazole (7-NI), a selective nNOS
inhibitor, in the consolidation of contextual fear memory in pigeons. The enzymatic activity of Ca2+-dependent and independent
NOS and protein expression of nNOS in the hippocampus were analyzed.
We used adult male pigeons with an average 350 g weight, distributed in 5 groups: 7-NI group (7-NI, 100nmol/0.5/l., i.c.v., n =
8), vehicle group (VEIC, DMSO 5% and PBS, n = 8), conditioning-untreated group (COND, n = 8), context-untreated group
(CONT, n = 8) and naive (NAIVE, n = 8). The pigeons were submitted to surgery for implant of an intracerebroventricular (i.c.v.)
cannula. Administration of 7-NI or vehicle followed immediately the 20 min training session which had three context-shock
pairings. After 24 hours, the animals were tested in a session of re-exposure to the conditioning context during 5 min. CONT
group was exposed to context without shock presentations and the NAIVE pigeons were only daily weighed. The sessions were
digitally recorded for posterior transcription and behavioral analysis. The hippocampal tissue was analyzed for enzymatic activity
of Ca2+-dependent and independent NOS which was expressed as pmols L-citrulline produced per minute and per mg of protein.
Protein expression of nNOS was analysed by the Western Blotting method and the values of optical densitometry of the
immunoreactive nNOS bands were normalized for the total protein content of the samples as determined by Ponceau S solution
for histochemical staining. In the training session of 7-NI, VEIC and COND groups there was greater expression of freezing in
the last five blocks of 1 min than in the first five blocks of 1 min (p < 0.05) and between 1 min interval blocks during the sessions
(p < 0.05). The enzymatic activity of Ca2+-dependent NOS showed marginally significant difference between groups (p = 0.06;
ANOVA). There was no significant difference in the enzymatic activity of Ca2 +-independent NOS (p > 0.05). Values of optical
densitometry of the immunoreactive nNOS bands in 7-NI, VEIC, COND and CONT groups were greater than the observed for
NAIVE group (p <0.05).
Conclusions:
The behavioral data indicate that inhibition of nNOS with i.c.v. administration of 7-NI immediately after training disrupted the
consolidation of aversive contextual memory in pigeons. The alteration of both the enzymatic activity of NOS and the
quantitative nNOS protein expression in the hippocampus suggests the participation of nitric oxide in the retrieval of aversive
contextual.
Keywords: contextual fear conditioning, hippocampus, neuronal nitric oxide synthase, 7-nitroindazol, freezing
Financial Support: FAPESP process 2009/13213-7
Resumo:21-107
CARDIOVASCULAR RESPONSES EVOKED FROM THE PERIAQUEDUCTAL GREY REQUIRE NEURONAL
ACTIVITY IN THE AMYGDALA.
de Abreu, A. R. R. ; Abreu, A. R. R. ; de Menezes, R. C. A

Departamento de Cincias Biolgicas, UFOP
Objectives:
The periaqueductal gray matter (PAG) is involved in the integration of specific cardiovascular changes associated with emotional
behaviors. Moreover, microinjection of excitatory aminoacids (EEA) into the lateral/dorsolateral column of the PAG (l/dlPAG)
evokes increases in the heart rate (HR) and in the mean arterial pressure (MAP) in a pattern similar to those observed during
emotional stress. This pattern of cardiovascular changes can also be attained by activation of the basolateral amygdala (BLA)
neurons. Therefore, the aim of our study is to evaluate the role of the BLA in the cardiovascular responses evoked by the
stimulation of PAG.
Methods: Male Wistar rats (300 20g) were anesthetized with ketamine-xylazine (80 mg/kg 11.5 mg/kg, i.p), and ipsilateral
guide cannulae were implanted into the l/dlPAG and into the BLA. In another set of rats we implanted guide cannulae into the
l/dlPAG and into a region at the vincinity of the BLA (NBLA), but outside of it (Negative Control). Six days latter, the rats were
anesthetized again (Isoflurane 2% in 3l of O2) and a catheter was inserted into the femoral artery for recording of MAP and HR.
Experiments were performed 48 hr later. Each rat was subjected to two different trials, 24h apart, and in random order, in which
either saline vehicle (100 nl) or muscimol (100 pmol/100 nl) was microinjected into the BLA/NBLA followed, 5 minutes later,
by the microinjection of a EEA, NMDA (6pmol/100nl), into the l/dlPAG. Results: Microinjection of muscimol into the BLA
attenuated the increase in HR evoked by the microinjection of NMDA into the l/dlPAG when compared with the saline treatment
(24 9 vs 79 10 bpm after saline, n=6, p=0,0003, by student paired t-test). Likewise, the microinjection of muscimol into the
BLA reduced the increase in MAP produced by the microinjection of NMDA into the l/dlPAG (6 2 vs 16 4 mmHg after
saline, n=6, p=0,021, by student paired t-test). In negative control animals, microinjection of muscimol into the NBLA did not
attenuate the increase in HR evoked by the microinjection of NMDA into the l / dlPAG compared with saline treatment (71 16
vs 88 5 bpm, n=4, p=0,3248, by student paired t-test). Similarly, the microinjection of muscimol into the NBLA did not reduce
the increase in MAP produced by the microinjection of NMDA into the l / dlPAG (13 3 vs 22 7 mmHg after saline, n=4,
p=0,3554, by student paired t-test).
Conclusions:
These results indicate that the neuronal activity in the region of the BLA plays a role in the generation of physiological response
induced by activation of the l/dlPAG neurons.
Keywords: Amygdala, Muscimol, NMDA, Periaqueductal, Stress
Financial Support: CNPq, FAPEMIG e UFOP
Resumo:21-108
BDNF AND ESTROGEN LEVELS ARE CORRELATED IN WOMEN WITH BIPOLAR DISORDER
Colpo, G. D. 1,2,3; Sulzbach, M. 2,3; Brietzke, E. 2,3; Tramontina, J. 2,3; Migliavacca, F. 2,3; Goi, P. D. 2,3;
Wollenhaupt - Aguair B. 1,2,3; Ceresr, K. M. M. 2,3; Sant'anna, M. K. 2,3; Kapczinski, F. 1,2,3
1
Programa de Ps Graduao Cincias Mdicas/ UFRGS, UFRGS
2
Programa de Transtorno Bipolar/ HCPA, HCPA
3
INCT Translacional em Medicina, INCT-TM
Objectives:
Hormonal changes throughout life are associated with mood changes in healthy women. Estrogen (E) seems to have a positive
correlation with the levels of brain-derived neurotrophic factor (BDNF) in healthy volunteers. However, this correlation was not
investigated in patients with bipolar disorder (BD). The study aims to investigate the relationship between brain-derived
neurotrophic factor (BDNF) levels and hormones involved in hypothalamus-pituitary-gonadal axis in women with bipolar
disorder (BD), during reproductive years and also in postmenopausal group. In addition, differences across the two phases of
menstrual cycle were also evaluated in the group during reproductive years.
Thirty two euthymic women with BD were included. Patients in reproductive years had regular menstrual cycles without
hormonal contraceptive and postmenopausal women were not under hormonal replacement therapy. Endocrine instable
conditions were considered an exclusion factor. Blood samples were withdrawn for measures of BDNF, estrogen (E),
Progesterone (P), LH and FSH levels, being collected in follicular and lutheal phases of menstrual cycle. Menopausal women
were held only one blood sample. Diagnoses were assessed using SCID-I. Results: Considering the whole sample, BDNF
presented a positive correlation with E levels (Pearson Correlation, coefficient= 0.36, P= 0.043). In patients in reproductive
phase, there was a negative correlation with FSH (Pearson Correlation, coefficient= 0.831, P= 0.040). A similar result was found
with LH levels (Pearson Correlation, coefficient= 0.908, P=0.012) in the second phase of menstrual cycle.
Conclusions:
There was a statistically significant positive correlation between BDNF and E levels, with higher BDNF levels being associated
with o estrogenic stimulation. The results of this study open new avenues of investigation in pathophysiology of mood
abnormalities related to hormonal variation as well as in their treatment.
Keywords: bipolar disorder, women, brain-derived neurotrophic factor , estrogen, hypothalamus-pituitary-gonadal axis
Financial Support: HCPA, CNPq
Resumo:21-109
RENOVASCULAR HYPERTENSION INFLUENCES THIRST AND SODIUM APPETITE IN RATS.
Blanch, G. T. 1; Freiria-oliveira, A. H. 1; Amaral, A. A. 1; Sumners, C. 2; Menani, J. V. 1; Colombari, E. 1;

Colombari, D. S. A. 1
1
Department of Physiology and Pathology, FOAr-UNESP
2
Department of Physiology and Functional Genomics, Univ. of Florida
Objectives:
Previous studies have shown that daily 2% NaCl intake was greater in rats with renovascular 2 kidney 1 clip hypertension [2K1C;
Schomig et al, Clin. Exp. Pharmacol. Physiol., 7(2): 169, 1980]. However, whether 2K1C hypertension affects induced water
intake and/or sodium intake is unknown. Thus, we aimed to study the effect of 2K1C hypertension on water intake induced by
increasing plasma osmolality (intragastric 2 M NaCl) and on sodium appetite induced by the treatment with furosemide (FURO)
along with a low dose of captopril (CAP).
Male Holtzman rats (150-180 g) maintained with water and 0.3 M NaCl ad libitum were anesthetized, and the left renal artery
was partially obstructed with a silver clip of 0.2 mm width. Sham animals were submitted to the same surgical procedure without
partial renal artery occlusion. After 5 weeks, on the day of the experiments, sham (n = 7) and 2K1C (n = 8) rats had water, NaCl
and food removed from the cages. The rats received, sequentially, 5 days apart, an intragastric (ig) load of 2 M NaCl (2 ml) and a
subcutaneous (sc) injection of FURO (10 mg/kg) + CAP (5 mg/kg). In both cases, urine output was collected over the next hour
after the treatments. After this 1 h period, the fluids were returned to the cage and intakes/outputs were recorded for 2 h. Urine
volume and natriuresis after hypertonic saline were similar amongst the groups (sham: 1.9 0.4, vs. 2K1C: 2.6 0.6 ml/1 h and
sham: 1874 179 vs. 2K1C: 1941 293 Eq/1 h). The water intake induced by ig administered 2 M NaCl was smaller in 2K1C
rats (sham: 9.5 0.8, vs. 2K1C: 6.2 1.1 ml/2 h, p< 0.05). In the FURO + CAP protocol diuresis (sham: 7.8 0.8 vs. 2K1C: 7.3
1 ml/1h) and natriuresis (sham: 853 80 vs. 2K1C: 752 124 Eq/1 h) values were not different between the groups.
Nevertheless, sodium intake induced by FURO + CAP was smaller in 2K1C rats compared to shams (sham: 5.9 0.9 vs. 2K1C:
3.0 0.4 ml/2 h, p<0.05).
Conclusions:
The results suggest that renovascular hypertension impairs water intake induced by hyperosmolality and sodium intake induced
by sodium depletion. Further studies are necessary to disclose the mechanisms involved with these responses in 2K1C rats.
Keywords: 2K1C, water intake, extracellular dehydration, cellular dehydration, diuresis
Financial Support: CAPES-PNPD, CNPq, FAPESP, NIH HL-076803
Resumo:21-110
A2 NEURON LESION INCREASES HYPOTHALAMIC MAGNOCELLULAR PARAVENTRICULAR NUCLEUS
ACTIVATION INDUCED BY PLASMA HYPEROSMOLALITY.
Freiria-oliveira, A. H. ; Blanch, G. T. ; Menani, J. V. ; Colombari, E. ; Colombari, D. S. A.

Dept of Physiology and Pathology, FOAr-UNESP
Objectives:
Plasma hyperosmolality activates the paraventricular (PVN) and the supraoptic nuclei (SON) of the hypothalamus and increases
vasopressin secretion. Previously we have shown that hyperosmolality causes vasopressin-dependent pressor response in rats
with specific lesions of the A2 noradrenergic group of the commissural nucleus of the solitary tract (A2/cNTS). In the present
study, using c-fos immunohistochemistry labeling, we aimed to investigate the neuronal activation in the hypothalamic areas
activated by plasma hyperosmolality in A2/cNTS-lesioned animals.
Male Holtzman rats (280-320 g) were submitted to lesions of dopamine-beta-hydroxilase (DH)-containing neurons in the
A2/cNTS achieved by injections of anti-DH-saporin (12.6 ng/60 nl, A2 lesion group, n=4) or sham lesions (injection of IgGsaporin, 12.6 ng/60 nl, n=4). Plasma hyperosmolality was induced by intragastric (ig) 2 M NaCl load (2 ml/rat). For training, the
animals were submitted to one ig 2 M NaCl load 7 days before the experiments and in the next 6 days, the animals received one
ig 0.15 M NaCl load (2 ml) daily. In the day of the experiments, half of the A2 lesioned and half of sham rats received ig load (2
ml) of 2 M NaCl and the remaining animals received ig load of 0.15 M NaCl. Two hours later the animals were deeply
anesthetized (pentobarbital 50 mg/kg ip) and perfused with 4% paraformaldehyde. C-fos imunohistochemistry procedure was
performed using DAB staining. Intragastric 0.15 M NaCl load promotes no c-fos expression in SON, magnocellular PVN
(mPVN) and parvocellular PVN (pPVN) in both sham and A2 lesion groups. Intragastric 2 M NaCl load similarly increased c-fos
expression in both SON and pPVN in sham animals (SON: 105 27 and pPVN: 22 9 cells/section) and in A2 lesioned animals
(SON: 88 7 and pPVN: 37 6 cells/section,). However, ig 2 M NaCl load increased more the c-fos expression in the mPVN of
A2 lesioned animals (90 13 cells/section, p < 0.05) compared to sham (47 20 cells/section).
Conclusions:
These results suggest that the activation of the mPVN by hyperosmotic stimuli is reduced or limited by an inhibitory mechanism
dependent on the cNTS A2 noradrenergic neurons.
Keywords: PVN, osmoreceptors, NTS, c-fos
Resumo:21-111
CANNABINOID CB1 RECEPTOR EXPRESSION IN THE RAT BASAL GANGLIA AND THE EFFECTS OF ACUTE
CANNABINOID TREATMENT IN RATS WITH 6-HYDROXYDOPAMINE-INDUCED HEMIPARKINSONISM
Chaves, G. P. ; Mazucanti, C. H. Y. ; Real, C. C. ; Britto, L. R. G. ; Torro, A. S.

Department of Physiology and Biophysics, ICB - USP
Objectives:
Although pharmacological evidence indicates the occurrence of cannabinoid CB1 receptors in the basal ganglia, detailed
morphological and functional data are scarce on this issue. Moreover, in Parkinsons Disease (PD) the function of CB1 and its
relation to other neurotransmitter systems (such as the GABAergic system) are both unclear. Yet the cannabinoid system seems
to have an important role in neurodegeneration and/or neuroprotection processes. We investigated here the possible colocalization of CB1 with markers for glial cells and structural elements of neurons and with tyrosine hydroxylase (TH) in the
basal ganglia of rats. We also analyzed, in 6-hydroxy-dopamine (6-OHDA)-induced PD rats, the GABA, GAD and CB1 levels in
the basal ganglia, the acute action of AM 404 (an inhibitor of anandamide reuptake) on the expression of TH and glial cell
markers, and the acute effects of different doses of ACEA (cannabinoid agonist) on the TH expression in the basal ganglia.
Double immunostaining of CB1 with markers for astrocytes (GFAP), oligodendrocytes (OD), microglia (OX-42), axons
(neurofilaments), dendrites (MAP-2) and TH was performed in the basal ganglia of normal rats. Other rats received injections of
6-OHDA (12g in 1l) in the right striatum, and the left striatum was used as a control. The 6-OHDA-lesioned animals were then
divided into five groups: (1) time course evaluation of the lesion after distinct survival times (3, 7, and 10 days); (2) acute
treatment with 2 mg/kg of AM 404 or (3) vehicle; (4) acute treatment with 0.5, 1, 2, and 4mg/kg ACEA or (5) vehicle. Brains
from the time course group were subjected to immunohistochemistry for TH, CB1, and GABA, and immunoblotting for GAD.
Brains of AM 404 group was processed for TH, GFAP, and OX42, and of the ACEA group was processed for TH
immunohistochemistry. The results were analyzed by one-way ANOVA or unpaired t test. We found scarce co-localization of
CB1 with markers of glial cells, but a marked co-localization with MAP-2, especially in the substantia nigra (SN) and the
external globus pallidus. The co-localization of CB1 with axon markers occurred primarily in the SN. Co-localization of CB1
with TH was not clearly detected in the basal ganglia. In PD-induced rats, no change was found in the expression of GABA or
GAD in basal ganglia at all survival times analyzed (N=3, p>0.05). On the other hand, a gradual decrease in CB1 staining of
about 30% and 40% in the SN was observed after 7 (N=4, p< 0.01) and 10 days (N=3, p< 0.05) after the lesion, respectively. The
treatment with AM 404 decreased the TH expression in SN by 10% (N=5, p
Conclusions:
This study adds information about the cellular localization of CB1 in the basal ganglia and indicates that CB1 receptor expression
decreases in PD by an event that is unrelated to the GABAergic system. It is possible that this event is related to some indirect
effect of the loss of dopaminergic neurons. Nevertheless, acute activation of CB1 appeared to be deleterious in the 6-OHDA rat
model of PD.
Keywords: Cannabinoid System, CB1 receptor, Neurodegeneration, Parkinsons Disease
Financial Support: FAPESP and CNPq.
Resumo:21-112
REACTIVE OXYGEN SPECIES AND ASCORBIC ACID ARE MODIFIED IN SPINAL CORD OF RATS WITH
NEUROPATHIC PAIN
Goecks, C. S. B; Horst,a. ; Moraes, M. S. ; Scheid, T ; Kolberg, C. ; Partata, W. A.

Fisiologia/ICBS, UFRGS
Objectives:
Reactive oxygen species (ROS) have been implicated in the development of persistent pain states, including neuropathic pain
(one ocurring as a result of peripheral or central nervous system injury). While the involvement of ROS in this condition is
becoming clear, the detailed changes to these species in different models of neuropathic pain are still unclear. Thus, this study
proposes to correlate hyperalgesia and hydrogen peroxide (H2O2), superoxide dismutase (SOD) and catalase activities, and
ascorbic acid (AA) measures in the spinal cord of rats submitted to chronic constriction injury (CCI) of the sciatic nerve, a useful
experimental model of neuropathic pain.
After approval of the trial by the Ethical Committee of Animal Research of the Federal University of Rio Grande do Sul (No.
2008052), adult male rats weighing 200-250 g were divided in three experimental groups (six animal/group): Nive (animals
which did not undergo surgical manipulation), Sham (animals in which all surgical procedures to expose the sciatic nerve were
used except the constriction of this nerve) and CCI (animals in which the sciatic nerve was exposed and 4 ligatures were placed
around the nerve at the proximal part of the trifurcation with a distance of 1 mm between each ligature). The animals were
sacrificed 3 and 10 days after the surgical procedure. The mechanical threshold with von Frey hairs and thermal latency with hot
plate were measured just before the surgery and 3 and 10 days after it. H2O2 determination was based on horseradish peroxidase-
mediated oxidation of phenol red. SOD activity was determined by the inhibition of superoxide radical reaction with pyrogallol.
Catalase activity was determined by following the decrease in H2O2. AA level was determined through the 2,4-dinitrophenylhydrazine derivative of dehydroascorbic acid. Data were expressed as means and errors and compared by two-way
ANOVA followed by the Bonferroni test. Differences were considered statistically significant when p
Conclusions:
Overall, these data demonstrate that hyperalgesia attributed to the CCI model of the sciatic nerve coincides with changes in spinal
cord antioxidant enzymes and AA. These modifications should be occurring to maintain the reduced levels of H2O2. However,
such reduction did not attenuate the hyperalgesia signs caused by CCI of the sciatic nerve.
Keywords: chronic constriction injury , sciatic nerve, superoxide dismutase, catalase, hydrogen peroxide
Resumo:21-113
PROFILE FUNCTIONALITY AND DEPRESSIVE STATE IN PATIENTS WITH PARKINSON'S DISEASE
Freire, Q. C. 1; Filho, H. D. A. G. 1; Lima, A. L. S. D. 1; Arago, C. D. A. 1; Camplo, C. L. D. C. 1; Neto,

E. R. 1; Campelo, M. D. G. L. D. C. 2; Franco, C. I. F. 1
1
Depto. de Fisioterapia/ Universidade Estadual da Paraba, UEPB
2
Depto. de Medicina/ Universidade Federal de Campina Grande, UFCG
Objectives:
Individuals with Parkinson's Disease (PD) may present cognitive decline, causing losses in thinking, perception and trial. Thus,
difficulties with memory, calculation and activities that require spatial orientation end up occurring with greater frequency. This
study aimed to evaluate motor and cognitive changes as well as depressive symptoms present in patients with PD.
METHODS: The sample consisted of eight individuals of both sexes with a clinical diagnosis of PD, assisted in Physical Therapy
Clinic of UEPB. The instruments used were the Neurological Assessment Protocol for socio-demographic, Rating Scale Unified
Parkinson's disease (UPDRS) for analysis of motor exploration activity, and the Beck Depression Inventory (BDI), to survey the
intensity depressive symptoms. Data were analyzed using Graph Pad Prism 4.02, and values are expressed in percentage, mean
and standard deviation, considering significant values with p
Conclusions:
Based on data obtained, it is possible to suggest that changes in PD motor precedence over the cognitive, as well as most of the
sample showed signs of depression.
Keywords: Cognition, depressive state, motor changes, Parkinson's disease
Financial Support: PROBEX / UEPB
Resumo:21-114
STROKE: ANALYSIS OF CORRELATION OF TRANSFER FUNCTION BETWEEN TWO INSTRUMENTS FOR
MOTOR EVALUATION.
Silva, M. J. C. ; Franco, C. I. F. ; Monteiro, D. L. ; Brito, A. S. S. ; Tavares, C. D. ; Arajo, D. P.

Depto. de Fisioterapia/Universidade Estadual da Paraba, UEPB
Objectives:
The hemiparesis is the classic clinical signs resulting from an ischemic or hemorrhagic cerebrovascular disease, involving cortex
or brainstem, which compromises the motor control of the affected hemisphere. Spasticity leads to loss of functional capacity and
personal achievement of these individuals. The aim of this study was to investigate the correlation between the transfer function
of two motor evaluation instruments in chronic hemiparetic.
This is an exploratory, descriptive and cross-sectional study involving 17 individuals assisted in the extension project "Group of
Interdisciplinary Assistance of the hemiparetic patient - GAIPH" conducted at the School of Physiotherapy, State University of
Paraba - UEPB. It was used as an evaluation tool, Neurological Assessment Protocol for characterization socio-demographic
data, the category transfer of Motor Functional Independence Measure (MIFm) and the Berg Balance Scale (BBS). The data were
analyzed using the software Graph Pad Prism 4.02, and values are expressed in percentage, mean and standard deviation,
considering significant values with p <0.05).
Conclusions:
Based on the results we suggest that there was no correlation between the transfer function of FIM and command of BBS, except
for field stationary tests.
Keywords: Berg Balance Scale, Cerebrovascular Disease, Hemiparesis, Motor Functional Independence Measure, Physiotherapy
Financial Support: PROBEX-UEPB
Resumo:21-115
SOCIAL ISOLATION IMPAIRED OLFACTION AND SOCIAL DISCRIMINATION MEMORY, BUT NOT
NEUROGENESIS IN THE MOUSE OLFACTORY BULB.
Monteiro, B. M. D. M. 1; Raslan, A. C. S. 1; Moreira, F. A. 2; Massensini, A. R. 1; Moraes, M. F. D. 1;

Pereira, G. S. 1
1
FISIOLOGIA/UNIVERSIDADE FEDERAL DE MINAS GERAIS, UFMG
2
FARMACOLOGIA/UNIVERSIDADE FEDERAL DE MINAS GERAIS, UFMG
Objectives:
To evaluate the effect of social isolation on olfaction and social discrimination memory, as well as on olfactory bulb
neurogenesis.
Methods: twenty C57/Bl6 male mice (8-12 weeks old) were divided in two groups: social isolated (SI) and group-housed (GH)
for 6 days. Then, each animal was placed in a box containing two empty cylinders with around 60 holes. After 30 min, one Swiss
juvenile (around 21 days old) was introduced into each cylinder and the time spent in social exploration (when the resident
animal introduced its nose and/or whiskers inside any of the cylinders holes) was quantified during 5 min. Twenty-four hours
later, the animal was re-introduced in the same box, now containing one familiar and one new juvenile. The social behavior was
measured during 5 min. As a control for olfactory perception, each mouse had one piece of chocolate introduced in its home cage
(7g/day during 3 days). Then, the food and chocolate were withdrawn and, 12 hours later, the animals were introduced in a new
box, where the latency to find a hidden piece of chocolate was measured. A second set of animals was used to measure
neurogenesis in the olfactory bulb. Ten Swiss male mice (8-12 weeks old) were divided in SI and GH groups and received a daily
injection of BrdU (75 mg / kg) during 7 days. Twenty-four hours after the last injection, they were perfused with 4%PFA and the
brains were processed for BrdU and NeuN immunofluorescence. Images were captured using a microscope and a Z-stack analysis
was performed at 1um intervals. New neurons were quantified using Image J software. The data were analyzed by Students t
test. Results: Both groups present similar social recognition indexes in the day 1 (GH=0.370.10; SI=0.390.11; p=0.74), but
only the GH group was able to discriminate juveniles in the day 2 (GH=0.660.1; SI=0.390.14; p=0.0002). The SI group took
longer to find the hidden chocolate compared to GH group (GH =20.166.74s;SI=46.5615.05s; p=0.0001). There was no
significant difference between groups in the number of BrDu/NeuN positive cells in the olfactory bulb.
Conclusions:
Social isolation impaired social discrimination memory, probably by reducing the animals olfactory capacity. However, the
decrease in the sensory input did not impaired cell proliferation in the olfactory bulb. Thus, it seems that the mnemonic and
sensorial changes caused by social isolation cannot be attributed to a decrease in neurogenesis.
Keywords: NEUROGENESIS, OLFACTION MEMORY, SOCIAL ISOLATION , SOCIAL DISCRIMINATION MEMORY
Financial Support: CAPES, CNPq, FAPEMIG
Resumo:21-116
ABSENCE OF TYPE I AND II INTERFERON SIGNALING AND THE EXPRESSION OF ANXIETY-LIKE
BEHAVIORS IN MICE
Saito, V. M. ; Rodrigues, D. H. ; Miranda, A. S. ; Teixeira, A. L.

Laboratrio de Imunofarmacologia, UFMG
Objectives:
The objective of this study was to evaluate the role of interferons (IFN) in the expression of emotional behaviors by submitting
nave female knockout IFN-/ receptor mice (IFNAR(-/-)) and IFN- deficient mice (GKO) and their wild type (WT) littermates
to behavioral paradigms commonly used to study depressive- and anxiety-like traits, i.e., the Elevated Plus Maze (EPM),
Porsolts Forced Swim Test (FST) and Open Field Test (OFT).
Female IFN- knockout mice (GKO; n=13), IFN-/ receptor knockout mice (IFNAR(-/-), n=8) and their wild type littermates
(WT; n=22) raised on a C57BL/6J genetic background (7 to 10 weeks old) were tested on Day 1 on the EPM, where the number
of open arm entries and time spent on open arms during a period of 5 minutes were recorded. On Day 2, all groups were tested on
the OFT, which consisted of a circular arena, divided into 4 central portions and 8 peripheral sectors where animals were allowed
to explore freely for 5 minutes and their locomotion score was recorded; and in the FST, in which mice were placed individually
in a 3-liter glass beaker filled with water and motion recorded for 6 minutes. Immobility time was measured during the last 4 min
of the session. Behavioral analyses were performed by a trained observer. Data were analyzed using one-way ANOVA followed
by Bonferronis post test. Values are expressed as Mean Standard Error of the Mean. A p value less than 0.05 was considered
statistically significant. In the EPM, the percent time spent in open arms averaged 34.09 5.2 in the control group (n=22).
IFNAR(-/-) mice spent significantly more time in the open arms (65.1 6.0, n=8, p<0.05).
Conclusions:
These findings suggest that IFN-gamma-deficient mice may have alterations in brain pathways that regulate behavior. The
complex phenotype displayed by KO mice indicates that these animals cope differently with stressors, and further studies are
necessary to investigate the biochemical aspects of this phenomenon.
Keywords: Anxiety, Behavior, Cytokine, Genetic knockout, Neuroimmunology
Financial Support: CAPES/FAPEMIG
Resumo:21-117
THE CO-CHAPERONE BAG2 REVERSES NICOTINE-INDUCED UP-REGULATION OF TOXIC TAU IN SH-SY5Y
CELL LINE - IMPLICATIONS FOR ALZHEIMER'S DISEASE.
Oliveira, A. S. A. D. 1,2; Costa, M. A. 2; Fior-chadi, D. R. 2; Carrettiero, D. C. 1

1
2
Departamento de fisiologia - IB/Universidade de So Paulo, USP
Objectives:
Alzheimer's disease (AD) is well characterized by the presence of histopathological hallmarks like neurofibrillary tangles (PHF)
composed mainly by intracellular hyperphosphorylated microtubule-associated protein Tau. A decrease in nicotinic receptor on
cellular membrane is also reported in AD patients. The co-chaperone BAG2 up-regulate the clearance of toxic Tau by increasing
cellular degradations process. In light of this, the aim of the present study is to investigate the relationship between nicotinic
receptor activation and the modulatory effect of BAG2 activity on hyperphosphorylated Tau clearance in SH-SY5Y cell line.
Human neuroblastoma SH-SY5Y cells were grown in a 1:1 mixture of essential growth medium DMEM and F12 supplemented
with 15% of fetal bovine serum, antibiotic and allowed it to grow at 37C. 24 h after plating, SH-SY5Y cells were treated with
different concentrations of nicotine (10, 50 and 100M) by 6h in the presence and absence of pEGFP-C1 expression vector
containing the cDNA sequence of BAG2. After treatment, the proteins were extracted and subjected to Western blotting
technique in order to investigate the levels of hyperphosphorylated Tau using P-Tau Ser 199/202 antibody. Superexpression of
BAG2 promoted a decrease in phosphorilated Tau levels (68.67 9.26*, % of control). In absence of exogenous BAG2
expression, nicotine promoted an increase in phosphorilated Tau levels (10M, 111.20 11.28; 50M, 137.90 9.30*; 100M,
153.40 10.68**, % of control). However, in the presence of superexpressed exogenous BAG2, nicotine promoted a decrease in
phosphorilated Tau levels (10M, 40.33 9.02**; 50M, 29.33 4.91**; 100M, 15.00 4.04**, % of control). Statistical
analyses were performed using one-way ANOVA followed by Dunnetts multiple comparison test. Values are shown as mean
S.E.M., *p < 0.05, **p < 0.01, n=3.
Conclusions:
The relationship among nicotinic receptor, hyperphosphorylated Tau and BAG2 is not yet characterized in the scientific
literature. We demonstrated in the present study that the co-chaperone BAG2 indeed up-regulate the clearance of toxic Tau.
However, the main point of the present work is that the co-chaperone BAG2 reverses nicotine-induced up-regulatoins of toxic
Tau. In other words, nicotinic receptor activation might increase BAG2 activity and consequentially up-regulate
hyperphosphorylated Tau clearance in SH-SY5Y cell line. These data suggest a possible pharmacological approach for
Alzheimer's disease.
Keywords: Alzheimer, BAG2, Degradation, Nicotinic receptor, Tau phosphorylation
Financial Support: UFABC, FAPESP
Resumo:21-118
MEMORY EVALUATION AND ANTIEPILEPTIC ACTION OF INTRA-HIPPOCAMPAL INJECTION OF
ADENOASSOCIATED VIRAL (AAV) GENE OF THE NEUROPEPTIDES Y (NPY) IN THE PILOCARPINE MODEL
IN MARMOSETS (CALLITHRIX JACCHUS)
Blanco, M. M. 1; Kohek, S. R. B. 1,1; Baldaia, M. A. 1; Pontes, J. C. C. 1; Cinini, S. M. 1; Mendes, P. P. 1;

Bland, Rj2; Fitzsimons, H. L. 2; During, M. J. 3; Vezzani, A. 4; Mello, L. E. 1
1
Fisiologia, unifesp
2
Neurology, Neurologix
4
Neuroscience, Laboratory Exp. Neurol., Mario Negri Inst
3
Molecular Virology,Immunology and Med Genetic, OSU
Objectives:
Here we evaluated the role of overexpression of NPY as an anticonvulsant and memory effects based on the administration of a
recombinant adeno-associated viral vector (rAAV) in marmoset.
Marmosets were subjected to stereotaxic surgery for injection of the rAAV in the hippocampus and implanted with a sensor for
telemetry recordings of electrocorticography (ECoG).The titer of the injected solution was 2.7x1013 vg/mL (rAAV1-CBANPY). In addition we also had a control non-injected group. All animals were submitted to memory tests in three periods
(baseline [after virus/before pilocarpine], latent [short after pilocarpine] and chronic [long after pilocarpine]) and to the induction
of status epilepticus (SE) by means of pilocarpine administration (250mg/kg, i.p.). ECoG and histological analysis were
evaluated. RESULTS: The baseline memory test for the T5 (hippocampus-dependent) task showed an improvement in the
performance of the rAAV1-NPY group when compared to control groups. The induction of SE in rAAV1-NPY did not differed
in the characteristics of SE except for phase IV, where rAAV-NPY animals had a shorter duration (60+28 min) than controls
(160+27 min). Memory tests in rAAV1-NPY groups at the latent and chronic periods showed an improved performance when
compared to baseline period. The onset of SRS (21+3.5 and 9+3.7 weeks) and its frequency (0.02+0.01 and 0.50+0.01
seizures/day) were markedly different between rAAV-NPY and control animals, respectively. In the rAAV1-NPY group there
was an increase in the expression of NPY in the hippocampal region as compared to controls.
Conclusions:
Memory tests demonstrated improved performance that was specific for hippocampus-dependent-task in rAAV1-NPY injected
animals. The rAAV-NPY in the marmoset model of pilocarpine did not lead to clear anti-SE effects. In the chronic period
however there was a long-lasting NPY over-expression in neurons and a marked decrease in the frequency of spontaneous
seizures.
Keywords: neuropeptides y, marmoset, epilepsy, memory, pilocarpine model
Financial Support: FAPESP and CNPq (Brazil)
Resumo:21-119
PERINATAL UNDERNUTRITION INDUCES A LONG-LASTING DEFICIT OF CELL PROLIFERATION IN THE
HIPPOCAMPUS.
Pinho, M. C. 1; Matos, R. J. B. 2; Orozco-slis, R. 3; Lopes-de-souza, S. 4; Manhes-de-castro, R. 5;

Bolaos-jimenez, F. 6
1
Depto. de Fisioterapia/Universidade Federal de Pernambuco, UFPE
2
Ncleo de Educao Fsica e Cincias do Esporte, UFPE/CAV
3
Irvine University, IU
4
Departamento de Anatomia/Universidade Federal de Pernambuco, UFPE
5
Departamento de Nutrio/Universidade Federal de Pernambuco, UFPE
6
UMR1280-Physiologie des Adaptations Nutritionnelles, INRA
Objectives:
Maternal malnutrition alters various maturational events in the brain of the offspring resulting in learning and memory deficits
that extend into adulthood. At the cellular level, learning and memory relay on the production of new neurons in the hippocampal
dentate gyrus (DG), but whether adult neurogenesis is affected by early malnutrition has not been completely explored. To get a
better insight into the cellular mechanisms that might underlie the deleterious effects of perinatal undernutrition on learning and
memory, here we examined the neurogenesis processes in the hippocampus of adult rats exposed to prenatal and/or neonatal
nutrient restriction.
Pregnant Spraguey-Dawley rats were fed either ad libitum (C) or were undernourished by reducing their daily food intake by
50% in relation to the C group during gestation and lactation (FR) or during the lactation period only (AdLib/FR). At 60 days of
age, the offspring from each group was injected with bromodeoxyuridine (BrdU), and sacrificed after 2 h, 1 week or 3 weeks.
The number of BrdU-labeled cells in the DG was significantly reduced in the offspring of FR dams in relation to controls at all
the time points examined. However, the proportion of new cells exhibiting a neuronal phenotype was higher in FR rats than in
controls as revealed by the co-labeling at 3 weeks of the BrdU-labeled cells with NeuN. AdLib/FR animals, exhibited also a
reduction in cell proliferation at 2h and 1 week. However, we found no significant differences at 3 weeks in either the number of
BrdU-labelled cells or in the proportion of new neurons between controls and AdLib/FR rats.
Conclusions:
These results indicate that undernutrition during perinatal development induces a long-lasting deficit of cell proliferation in the
hippocampus without affecting the neurogenesis process per se.
Keywords: cell proliferation , hippocampus, neurogenesis, Perinatal undernutrition
Financial Support: INRA-Universit de Nantes (France) and FACEPE/CNPq.
Resumo:21-120
SELECTIVE LESION OF RETROTRAPEZOID PHOX2B-EXPRESSING NEURONS ATTENUATES THE CENTRAL
CHEMOREFLEX IN CONSCIOUS RATS.
Ragioto, D. A. M. T. 1; Falquetto, B. 1; Colombari, E. 3; Moreira, T. S. 1; Takakura, A. C. 1

1
Department of Physiology and Biophysics, ICB USP
2
Department of Pharmacology, ICB USP
3
Department of Physiology and Pathology, FOAr UNESP
Objectives:
The retrotrapezoid nucleus (RTN) contributes to an unknown extent to the central chemoreflex (the activation of breathing by
elevation of CNS PCO2). Neurophysiological and genetic evidence suggests that the RTN neurons involved in this reflex are a
group of chemosensitive glutamatergic interneurons that express the transcription factor Phox2b and lack tyrosine-hydroxylase
(henceforth called RTN Phox2b+TH neurons). In the present study, we ask whether the selective destruction of Phox2b+THRTN neurons could affect breathing in conscious rats.
Male Wistar rats (280-300 g) with bilateral injections of the toxin [Sar9,Met(O2)11]-substance P (SSP-SAP) were used. We
showed that RTN contains around 2000 Phox2b+TH- cells and bilateral injections of SSP-SAP into RTN destroyed Phox2b+THneurons but spared facial motoneurons, catecholaminergic and serotonergic neurons and the ventral respiratory column caudal to
the facial motor nucleus. Bilateral inhibition of RTN neurons with SSP-SAP (0.6 ng/30 nl) reduced resting ventilation
(1029142, vs. saline: 1663137 ml/min/kg) and the increase in ventilation produced by hypercapnia (7% CO2) (2916162, vs.
saline: 3736243 ml/min/kg). In anesthetized rats with bilateral lesions of around 80% of the Phox2b+TH- neurons, acute
activation of the Botzinger or the preBotzinger complex with NMDA (5 pmol/50 nl) elicited normal phrenic nerve activity.
Conclusions:
In conclusion, the destruction of the Phox2b+TH- neurons is a plausible cause of the respiratory deficits caused by injection of
SSP-SAP into RTN. Our results also suggest that RTN neurons activate facilitatory mechanisms important to the control of
ventilation in resting or hypercapnic conditions in conscious rats.
Keywords: hypercapnia, retrotrapezoid nucleus, Phox2b, central chemoreceptors, breathing
Resumo:21-121
CORRELATION OF FEAR OF FALLING ON THE BALANCE OF INDIVIDUALS WITH CHRONIC
CEREBROVASCULAR ACCIDENT (CVA).
Silva, C. A. D. ; Monteiro, K. S. ; Souza, C. G. D. ; Pontes, I. E. D. A. ; Lima, J. D. ; Silva, P. G. E. ;

Franco, C. I. F.
Depto.de Fisioterapia/Universidade Estadual da Paraba, UEPB
Objectives:
The aim of this study was to correlate the fear of falling and the balance in individuals with chronic CVA.
17 individuals of both sexes with a clinical diagnosis of chronic CVA (= or> 6 months) assisted in the Clnica Escola de
Fisioterapia da UEPB were part of this research. Three instruments were used for methodological purposes: the Neurological
Evaluation Protocol for socio-demographic and clinical characterization, the Fear of Falling Scale (RMS) to assess fear of falling
in individuals and the Berg Balance Scale (BBS) to evaluate balance. Data were analyzed using the Graph Pad Prism 4.02, and
values are expressed in percentage, average and standard deviation, considering significant values with p
Conclusions:
Based on the results it is possible to suggest that the greater the fear of falling, lower is the balance in individuals with CVA.
Keywords: Cerebrovascular Accident, Balance, Fear Of Falling
Financial Support: UEPB
Resumo:21-122
THE EFFECTS OF AMBIENT TEMPERATURE ON SOCIAL INTERACTIONS IN RATS
Ishikawa, D. ; Almeida, M. C.
CCNH / Universidade Federal do ABC, UFABC
Objectives:
The mechanisms responsible for ambient temperature (Ta) detection although known to be essential for adequate
thermoregulation, are still not completely understood. The temperature influences not only physiological mechanisms, but is also
capable of influencing psychological aspects, behavior and wellness (J Pers Soc Psychol 17: 92-98, 1971; Am J Physiol Regul
Integr Comp Physiol 00377.02007, 2007.; J Pers Soc Psychol 15: 240-244, 1970; Science 322: 606-607, 2008). This study aims
to investigate the effect of different Ta on social behavior in rats.

We used 16 male Wistar rats weighing 290-400g. Rats were acclimated to different Tas [divided in cold (18C), cool (22C),
termoneutral (26C), warm (29C) and hot (32C)] for 2 hours. Thereafter they were placed in pairs and the behavior was
registered. The time spent on social interactions (sniffing, approaching or following the other animal) as well as the grooming (a
behavior associated to thermoregulation) were analyzed. Rats exposed to 22C (cool Ta) spent 49.8 4% of their time in social
interaction (n = 4 pairs). This time was slightly bigger in rats exposed to neutral (26C, n = 8 pairs) or warm Ta (29C, n = 3
pairs) (~ 52% in both groups). Time spent in social interaction at cold and hot Ta was 56 4 and 45.6 2%, respectively (n = 8
pairs for each group). While rats at neutral Ta spent only 3% of their time in grooming behavior, rats exposed to hot Ta spent
almost 10%.
Conclusions:
Our data suggests that warmer temperatures increases social interaction in comparison to cool Tas. The result that cold and hot
temperature respectively increases and decreases social interaction seems controversial, however, at these temperatures, animals
spent time in behavioral thermoregulation such as grooming (hot Ta) and most probably grouping (cold Ta). In fact, the
development of these behaviors (social interaction and thermoregulatory behaviors) are competitive, and from the moment that
maintaining the temperature becomes the most important motivation for the individual, the thermoregulatory behavior prevails at
the expense of social behavior, which explain the apparently controversial result.
Keywords: Ambient temperature, Grooming, Social interaction, Thermoregulation
Financial Support: PIC/UFABC (to D.I.), CNPq Universal (to M.C.A.)
Resumo:21-123
EVALUATION OF THE NEUROPROTECTIVE EFFECT OF CAFFEIC ACID IN MICE AFTER PERMANENT
BRAIN FOCAL ISCHEMIA.
Fernandes, F. D. P. ; Carmo, M. R. S. ; Neves, J. C. S. ; Fonteles, A. A. ; Nunes, A. C. L. ; Andrade, G. M.

Objectives:
According to the World Health Organization, occurs about 6 million deaths per year related to brain ischemia, representing the
third leading cause of death after coronary heart disease (7,2 million) and cancer (7,1 million). Brain stroke is the leading cause of
neurological disability that requests rehabilitation and care in industrialized countries (BROUNS, 2009). Studies of phenomena
related to the mechanisms of ischemia and hypoxia aimed at improving the quality of life of these patients.
There were 3 groups of animals: Sham-operated, Ischemic induced and Ischemic induced treated with caffeic acid. The shamoperated and the ischemic induced groups were were treated with drug vehicle (DMSO 20%) in the same way used in the
treatment of animals which were treated with the caffeic acid. The treated animals received two doses of 30mg/kg,
intraperitoneally, the first one 30 minutes before surgery, and the second one 1 hour after it. They were treated with this dose
twice daily for 3 days. The analyzed parameters were sensorimotor, acquisition and retention of memory and damaged cerebral
area by stroke. After ischemia, it was found through neurological assessment, a significant decrease in sensory function and
motor performance of animals. The percentage of infarct area in the treated animals was significantly lower than those submitted
to ischemia (SO = 0.89 0.18%; ISC = 9.06 1.2%; ISC + Caf.Ac. = 2.46 0.2%). It was also observed an increase in vertical
exploratory activity (number of Rearings) in treated animals compared to animals in group ischemic (ISC: 9.5 1.8, ISC +
Ac.Caf = 15.8 0.5). The treatment significantly improved the deficits in working memory induced by ischemia, assessed at Ymaze test (SO = 73.8 1.9%; ISC= 56.7 2.9%; ISC + Ac.Caf = 70.7 3.6%). Another similar results were observed in the
passive avoidance test in which the treatment improved acquisition of short memory and significantly long duration (SO = 235.0
33.3 s; ISC = 92.5 25.0 s; ISC + Ac.Caf = 296.7 1.85 s).
Conclusions:
The results demonstrate the caffeic acids antioxidants properties in the pathophysiology of cerebral ischemia, which showed a
significant effect neuroprotective on neuronal damage, motor behavior and memory after ischemia and indicate that this effect
may be related with the caffeic acids antioxidant properties.
Keywords: Caffeic Acid, Brain ischemia, Memory, Antioxidant effect, Neuroprotection
Resumo:21-124
NEUROTOXIC EFECTS OF 3-ACETYL-PYRIDINE ON HIPPOCAMPAL DENTATE GYRUS CITOLOGY AND
SPATIAL BEHAVIOR.
Araujo, V. M. 1; Rocha, E. D. 1; Freitas, F. S. 2; Rocha, M. S. 2; Allodi, S. 1; Cavalcante, L. A. 1

1
Instituto de Biofsica Carlos Chagas Filho/UFRJ, IBCCF
2
Instituto de Cincias Biomdicas/UFRJ, ICB
Objectives:
The hippocampal dentate gyrus is a region of the brain where the production of neurons throughout adult life, even in normal
conditions, has been best characterized. However, little is known about the properties of hippocampal glial cells or their response
to lesion. The objectives of the present work are to study the chronology of the morphological and phenotypical markers of
neuron and glial cells of the dentate gyrus in response to lesions induced by the neurotoxin 3-acetyl-pyridine (3AP), and to
investigate possible changes in spatial memory.
Thirty to thirty five day-old Wistar rats were submitted to a single 3AP intraperitoneal injection and euthanized 24 hours (h) or 4,
10, 21 or 36 days (d) after the lesion. Animals with 11, 21, and 36 d after 3AP and aged-matched controls were tested for changes
in spatial learning and memory using the Morris water maze. Coronal sections of the hippocampus from animals with 24 h to 21
d survival and their respective controls were processed for immunohistochemical reactions and the images were analyzed by
conventional microscopy. Digital images were collected and used for counting the cells in the hylus of the dentate gyrus and
performance in the Morris aquatic maze was evaluated. There was a decrease in microtubule-associated protein (MAP2)
immunoreactivity at 24 h and 4 d after 3AP treatment, as reported in the early stage of various hippocampal lesions. The most
significant changes were an increase in the number of hylus cells expressing GFAP at 24 h, suggesting that either differentiated
astrocytes are involved, together with microglia, in central nervous system responses from injury or that GFAP-expressing
neuroblasts are increased. The latter is reinforced by an increase in cells expression Beta 3-tubulin at 4 days. Spatial behavior was
significantly impaired at 11 d when compared with controls. This tendency still persisted at both 21 d and 36 d but was no longer
significantly different from the controls, suggesting that impairment of spatial learning and memory was reversible with time.
Conclusions:
These evidences suggest that damage to the hippocampus by the neurotoxin 3AP persists after recovery of the locomotory ability
but is, at least, partially reversible in young adult rats. Behavioral recovery is coherent with the time course of the progressive
changes in neurogenesis and synaptogenesis in this species.
Keywords: 3-ACETYL-PYRIDINE, glial cells, hippocampus, spatial behavior
Resumo:21-125
REACTION TIME AND ARTIFICIAL LIFE
Feher-da-silva, C. ; Baldo, M. V. C.
Instituto de Cincias Biomdicas - Universidade de So Paulo, ICB-USP
Objectives:
In a simple reaction time (RT) task, the subject has to respond as fast as possible to the abrupt appearance of a target stimulus
preceded by a cue, which may be valid (when it indicates correctly where the target will appear), neutral (when it doesn't indicate
where the target will appear) or invalid (when it indicates incorrectly where the target will appear). Performance is evaluated by
measuring the time elapsed between the appearance of the target and the subject's response. It is assumed that the cue directs
attention to the location it indicates and speeds up sensory processing there, so that for humans and other animals RT is lowest
when the cue is valid and highest when it is invalid. We used artificial life models to investigate the role of noise, action
selection, and neural processing capacity in the evolution of attentional mechanisms engaged by RT tasks.
We performed artificial life simulations wherein populations of artificial animals with limited processing capacity evolved
through a genetic algorithm based on their performance in a simple RT task. The proportion of valid, neutral, and invalid cues
was 8:5:2 or 8:15:2. When the simulation was run for 150 generations, the animals achieved the lowest possible RT (1 unit)
regardless of cue validity and made no errors. When the simulation was cut short at 30 generations, the animals had already
achieved very low RT averages (2.5-4.0 units) and error rates. The lowest RT was achieved for the most frequent cue type (the
frequency effect), whether it was valid or neutral, which indicates the animals were not using the information provided by the
cues. With cue proportion set as 8:15:2, the addition of gaussian noise (mean = 0, standard deviation = 2) to each neuron in the
neural networks caused a large increase in RT averages (RT: 31.9 .6, 30.4 .4, 34.1 .7 for the valid, neutral and invalid cues
respectively) and in errors, but the lowest RT was still achieved for the neutral cue. The task the animals had to perform was
switched to a choice RT task, wherein there were two possible responses, but only one was correct, depending on the location
where the target appeared. Cue proportion was 8:15:2. The results were similar to previous ones without noise, but the addition of
gaussian noise (mean = 0, standard deviation = 2) caused animals to respond fastest for the valid cue, even though the neutral cue
was the most frequent (RT: 37.7 .7, 40.4 .6, 63.8 4.2; p < .05). By equalling the number of neutral cues to the number of
valid cues and then to the number of invalid cues, the frequency effect was eliminated and the effect of the valid and invalid cue
relative to the neutral cue could be calculated: -5.3 or -12.5% and 10.6 or 21.3% respectively. Increasing the number of neurons
in the neural networks and thus their processing capacity didn't abolish the attentional effect, but instead increased the effect of
the valid cue.
Conclusions:
In artificial life models with noise and action selection, artificial animals were similar to humans and other animals in a RT task.
Our results suggest that noise and action selection, but not limited capacity, were important environmental features that helped
drive the biological evolution of attentional mechanisms engaged by RT tasks.

Keywords: attention, artificial life, reaction time, neural networks, noise
Resumo:21-126
EXPOSURE TO CIGARETTE SMOKE CONTAINING EITHER HIGH OR LOW LEVELS OF NICOTINE DURING
ADOLESCENCE
Guthierrez, M. C. S. 1; Manhes, A. C. 1; Filgueiras, C. C. 1; Cavina, C. C. 1; Naiif, V. F1; Ribeiro-carvalho,

A. 2; Abreu-villaa, Y1
1
Universidade do Estado do Rio de Janeiro, UERJ
2
Instituto Federal de Educao, Cincia e Tecnologia do Rio d, IFFRJ
Objectives:
Smoking typically begins during adolescence. Nicotine is an important psychoactive substance present in tobacco and, in fact, an
increasing number of animal studies on effects of nicotine in the adolescent brain is made available every year. However, tobacco
smoke contains about 4500 additional components. Accordingly, the possibility that tobacco compounds other than nicotine
participate in the effects of tobacco smoke can be raised. Despite that, there is a lack of experimental studies that investigate the
effects of tobacco smoke exposure during adolescence. The current study was undertaken to investigate effects of distinct levels
of adolescent nicotine exposure when combined to tobacco smoke on the behavior of mice. To do that, we generated smoke from
2 types of cigarettes, one containing a high level of nicotine and another containing a very low level of nicotine.
From postnatal day (PN) 30 to 45, Swiss male and female mice were exposed to tobacco smoke (whole body exposure for
8hr/day, 7days/week) generated from one of two reference research cigarettes: type 2R1F (HighNIC group nicotine=1,74mg/cig; n=37) or type 4A1 (LowNIC group - nicotine=0.14 mg/cig; n=19) in a chamber that received the smoke
generated from an automatic cigarette smoking machine. A smoke mixture containing 89% sidestream and 11% mainstream
smoke as a surrogate for active smoking, was generated at the rate of a single 35ml puff of 2s/min. Control animals (CT, n=30)
were exposed to air in a chamber like the chamber of smoke exposure. On PN44, 74 animals were submitted to the Elevated Plus
Maze (EPM) test. The EPM consists of two open (no walls, 5cm29cm) and two closed (5cm29cm15cm) arms, arranged
perpendicularly, and elevated 50cm above the floor.The percentage of time spent in the open arms of the maze was used as a
measure of anxiety, while the number of entries in the open+closed arms was used as a measure of activity. On PN45, the animals
were tested in the Hole Board which consists of a polyethylene box (30x38x20cm). The floor is divided into 16 rectangles with a
1.5cm diameter hole located on the center of each rectangle. The number of head-dips into the holes was used as a measure to
novelty seeking behavior. On PN45, cotinine (nicotine metabolite) serum levels from HighNIC, LowNIC and CT mice (n=12)
were determined using a kit from Orasure Technologies. (Bethlehem, PA). On PN45, HighNIC cotinine serum levels were 109.1
24.0ng/ml while LowNIC and CONT mice presented values below the detection limit of the technique (
Conclusions:
These results indicate that exposure to cigarette smoke alters the novelty seeking behavior, and the pattern of change depends on
the type of cigarette. Since exposure to smoke containing low concentrations of nicotine caused a reduction of the search for
novelty while exposure to smoke containing high concentrations of nicotine caused an increase of the search, we suggest that
among the constituents of cigarette smoke, nicotine is one of the determinants of our results. Additionally, this behavior has been
associated with a greater tendency to use drugs in humans, so, this model may be useful to investigate the susceptibility to drugs
of abuse.
Keywords: CIGARETTE , ADOLESCENCE, BEHAVIOR
Financial Support: Fellowships from CNPq; PIBIC-CNPq and CAPES and a grant from CNPq
(476991/2010-2
Resumo:21-127
EFFECTS OF NEONATAL EXPOSURE TO ORGANOPHOSPHATE PESTICIDES ON THE CENTRAL
CHOLINERGIC SYSTEM OF MICE
Nunes-freitas; A. L. 1; Lima; C. S. 1; Ribeiro-carvalho, A. 1,3; Dutra-tavares, A. C. 1; Nunes F. 1; Filgueiras,

C. C. 1; Manhes, A. C. 1; Meyer, A. 2; Abreu- Villaa Y. 1
1
UNIVERSIDADE DO ESTADO DO RIO DE JANEIRO, UERJ
2
UNIVERSIDADE FEDERAL DO RIO DE JANEIRO, UFRJ
3
INSTITUTO FEDERAL DO RIO DE JANEIRO, IFRJ
Objectives:
In countries in which economy is based on agribusiness, like Brazil, the large utilization of pesticides represents an important
problem of public health. Organophosphates (OPs) are among the most used pesticides worldwide. Classically, OPs act by
irreversibly inhibiting the enzyme acetylcholinesterase (AChE), the enzyme responsible for the breakdown of acetylcholine.
However, they may differ substantially in many of their noncholinesterase effects. Considering that previous studies describe that
OPs elicit more severe effects during development, here, we investigated whether early exposure to chlorpyrifos and
methamidophos at doses well below the threshold for systemic toxicity and that cause only 20% of acetylcholinesterase (ACHE)
inhibition elicit alterations in other biomarkers of cholinergic function.
From the 3rd to 9th postnatal day (PN), 80 Swiss mice were exposed to daily injections (s.c.) of either one of two different OPs,
methamidophos (METH; N= 10 females and 10 males) or clorpyrifos (CLOR; N= 10 females and 10 males) at doses (1 and
3mg/kg respectively) which were previously shown to cause low and similar AChE inhibition in brain (20%). Control animals
(CONT METH; N= 10 females and 10 males and CONT CLOR; N= 10 females and 10 males) received DMSO (s.c.). All
animals were sacrificed one day after the end of exposure, at PN10. We assessed the binding of [3H]hemicholinium-3 (HC-3) to
the high-affinity presynaptic choline transporter and choline acetyltransferase (ChAT) activity in the cerebral cortex and
brainstem. Results were evaluated by analyses of variance on all factors: treatment and sex. Treatment effects and interactions
were followed by post-hoc analyses using Fishers Protected Least Significant Difference (FPLSD). Cholinergic alterations
were sex-dependent, being significant only in females: In the brainstem, methamidophos promoted an increase in HC-3 binding
when compared to its corresponding control group (METH vs CONT METH, 46.7 2.1 vs 35.2 1.7; P=0,0006) In the
cortex, both clorpyrifos (CLOR vs CONT CLOR, 44.0 1.9 vs 34.6 2.1, P=0,0027, FPLSD) and methamidophos (METH vs
CONT METH, 49.0 2.5 vs 42.1 1.2; P=0,022, FPLSD) caused an increase in HC-3 binding. ChAT activity was not
affected.
Conclusions:
Considering that the high-affinity choline uptake system, assessed by HC-3 binding, is regulated by nerve impulse activity,
changes in HC-3 binding observed here represent an increase in cholinergic activity. In addition, since the doses of
methamidophos and chlorpyrifos used in the present study elicit similar levels of ACHE inhibition, the fact that these pesticides
elicited region-dependent effects, suggests that OPs can act through multiple mechanisms, not necessarily related to AChE
inhibition, that culminate in other cholinergic alterations.
Keywords: ORGANOPHOSPHATE, DEVELOPMENT, CHOLINERGIC SYSTEM, METHAMIDOPHOS, CLORPYFOS
Financial Support: CAPES, CNPq, FAPERJ and Mount Sinai school of Medicine
Resumo:21-128
SOMATIC MATURATION AND REFLEX ONTOGENY IN NEONATE RATS FED DURING PREGNANCY AND
LACTATION WITH A HIGH FAT DIET
Santos, T. D. ; Ferreira, A. K. ; Ladislau, H. F. L. ; Pereira-da-silva, M. S. ; Rocha-de-melo, A. P. ; Borba,

J. M. C.
Depto. de Nutrio - Lab. de Fisiologia da Nutrio -LAFINNT, UFPE
Objectives:
Aim: Investigate if a high fat diet intake containing 14% of hydrogenated vegetable fat during pregnancy and lactation influences
the somatic maturation and reflex ontogeny in neonatal rats.
Methods and Results: Female Wistar rats were fed a diet containing 7% soybean oil (control group - C, n = 32) or 14%
hydrogenated vegetable fat (Experimental Group - E, n = 47) during gestation and lactation. The pups were weighed at 1 st, 7th,
14th and 21st days of life. Somatic Maturation Indexes - eye opening (AO), ear unfolding (EU), auditory conduit opening (ACO),
eruption of the upper incisors (EUI), eruption of the lower incisor (EUI) and time of appearance of reflexes (palmar grasp - PG,
cliff avoidance - CA, vibrissae placing - VP, negative geotaxis - NG, righting - R, auditory startle response - ASR and free fall
righting- FFR) were evaluated in each litter (6 males and 2 females) daily between 8:00 and 10:00 am from the 1 st to 21st days of
life. Rats of the E group showed a lower weight gain at the 1 st lactation day (6 1; median interquartile) when compared to the
C group (6,82 - 0,852). This fact was also observed at the 7 th day of lactation (E: 14 2,75 and C: 15,83 4,9925) . The E group
showed a significant delay in the following indices (days): EU (3 -1, median interquartile) and ACO (13 1) compared to the
C group (EU: 3 0 and ACO: 12 1). The development of PG, CA and ASR was delayed in the E group (PG: 6 1, CA: 7 3,
ASR: 13 2,5) when compared to the C group (PG: 4 2, CA: 5 2, ASR: 12 0,25). On the other hand, R and FFR appeared
earlier in the E group (R: 4 1 e FFR: 15 0,5) when compared to the C one (R: 5 2 e FFR: 17 1).
Conclusions:
Conclusions: The chronic intake of a high fat diet, based on hydrogenated vegetable fat, seems to produce weight changes and
cause alterations in somatic and reflex maturation.
Keywords: High Fat, Hydrogenated Vegetable Fat, Rats, Reflex Ontogeny , Somatic Maturation
Financial Support: UFPE/ PROPESQ
Resumo:21-129
SURVIVAL OF RGC MEDIATED BY IL-2, BUT NOT BY IL-4, IS DEPENDENT OF HEPARAN SULFATE
EXPRESSION IN VITRO
Marra C. ; Moret, D. G. ; Corra, A. D. S. ; Sholl-franco, A.

IBCCF - Universidade Federal do Rio de Janeiro, IBCCF - UFRJ
Objectives:
Programed cell death (PCD) during development is widespread in both vertebrates and invertebrates, occurring in virtually all
cell types of the nervous system, including retinal neurons such as retinal ganglion cells (RGC). In addition to physiological
conditions, this phenomenon also occurs in retinal pathologies as observed in optic neuropathies, a group of retinal diseases
characterized by RGC loss visual field reduction and/or blindness blindness. Several factors are involved in the regulation of
RGC death, including cytokines and extracellular matrix (ECM) molecules. Previously, we have demonstrated the
neuroprotective effect of interleukin-2 (IL-2) and interleukin-4 (IL-4) upon retinal ganglion cells in vitro. In this work we: (i)
investigated if cytokines treatment could regulate the expression of heparan sulfate proteoglycans (HSPG) in retinal tissue, and
(ii) analyzed the correlation between the neuroprotective effect of cytokines and the heparan sulfate (HS) expression.
Newborn rats were anaesthetized by hypothermia and received injections of rhodamine dextran (RD) in their superior colliculus
for retrograde staining of RGC. After 2 days, retinal explants were obtained and maintained with or without IL-2 (50U/mL), IL-4
(5U/mL) and/or heparitinase for 5 days in vitro (DIV). The total number of nuclei (stained with DAPI) in the ganglion cells layer
and the specific number of RGC labeled with RD were counted using fluorescence microscopy.Immunohistochemical analysis
were performed to verify the HS expression in retinal tissue. The results showed that IL-2 and IL-4 treatment of retinal explants
increased the HSPG expression in all retinal layers after 5 DIV. Moreover the neuroprotective effect of IL-2, but not IL-4 was
completely blocked by pre-treatment with heparitinase.
Conclusions:
Our data demonstrate that IL-2 and IL-4 modulates the expression of HSPG in retinal tissue, and that early presence of this
molecule is essential for RGC survival mediated by IL-2, but not by IL-4.
Keywords: Retina, Retinal Ganglion Cells, Citokines, Extra Celular Matrix, Heparan Sulfate
Financial Support: CNPQ , CAPES, FAPERJ
Resumo:21-130
ETHANOL-INDUCED LOCOMOTOR SENSITIZATION INCREASES FOSB IMMUNOREACTIVITY IN THE
HIPPOCAMPUS AND AMYGDALA OF ADULT SWISS MICE.
Coelhoso, C. C. 1; Filev, R. 1; Varela, P. 1; Engelke, D. 1; Mello, L. E. 1; Santos-junior, J. G. 1,2

1
Departamento de Fisiologia/Universidade Federal de So Paulo, UNIFESP
2
Departamento de Cincias Fisiolgicas/FCMSCSP , FCMSCSP
Objectives:
Chronic exposures to drug of abuse induce several neuroadaptations in neuronal circuitry related to addiction. One of them is the
increase in FosB expression, notably in the dorsal striatum. Here, we investigate the FosB immunoreactivity in mice sensitized
and non-sensitized to the stimulant locomotor effects of chronic ethanol exposures.
Male Swiss mice (90 days, 30-40 g) were maintained for 15 min in an activity box and the distance traveled during this period
was measured. After that, the animals (N=32) were treated with ethanol (2 g/kg, i.p. 15% v/v) for 21 consecutive days. Control
animals (N=8) were treated with saline. After 4 days of withdrawal, the animals were challenged with ethanol (1.4 g/kg, i.p.) and
again placed in the activity box. In the next day, the animals were perfused and their brains processed for FosB/FosB
immunohistochemistry. According to the locomotor activity in the challenge day, ethanol treated animals were distributed in two
sub-groups: sensitized (1 SD above the mean) and non-sensitized (1 SD below the mean). FosB/FosB immunoreactivity was
measured in the medial prefrontal cortex (mPFC), nucleus accumbens core (NAco) and shell (NAsh), caudate-putamen (CPu),
hippocampus (HPC), ventral tegmental area (VTA), basolateral (BlA) and central nucleus of the amygdala (CeA). The data were
analyzed by one-way ANOVA followed by Tukey post hoc when necessary. The significance was set at PP=0.16]. Moreover,
Et_Sens had a higher locomotor activity after challenge when compared to the other groups [F(2,21)=57.93, PPPPP<0.001].
Conclusions:
These results suggest that increases on FosB/FosB in HPC (DG, CA1 and CA3) and in CeA could play an important role in the
ethanol-induced locomotor sensitization in mice.
Keywords: Ethanol, FosB , Immunohistochemistry, Locomotor Sensitization
Resumo:21-131
TEMPORAL BONE MODIFICATIONS FOLLOWING SPINAL CORD INJURY IN RATS.
Medalha, C. C. 1; Amorim, B. O. 1; Rosseti, K. 1; Ribeiro, D. A. 1; Pereira, R. M. R. 2; Renn, A. C. M. 1

1
Departamento de Biocincias, UNIFESP
2
Departamento de Reumatologia, USP
Objectives:
The aim of this work was to investigate the temporal alterations on bone mass, bone biomechanical properties and bone
morphology in spinal cord injured rats 2, 4 and 6 weeks after surgery.
Fourty male Wistar rats (aged 8 weeks and weighing 290 6.8 g) were randomly distributed into four groups (n=10 each group):
sham control group (CG: control without spinal cord-injured animal); spinal cord-injured 2 weeks (2W: spinal cord-injured
animals sacrificed 2 weeks post-surgery); spinal cord-injured 4 weeks (4W: spinal cord-injured animals sacrificed 4 weeks postsurgery); spinal cord-injured 6 weeks (6W: spinal cord-injured animals sacrificed 6 weeks post-surgery). The animals were
anesthetized by an intraperitoneal injection of ketamine (90 mg/kg) and xilasine (10 mg/kg) and a laminectomy was executed at
Th9-10. In injured rats, the duramater was exposed and the spinal cord was completely sectioned with microscissors. To reveal
temporal changes to the locomotor function after SCI, an evaluation was carried out by using the Basso, Beattie and Bresnahan
(BBB) Locomotor Rating Scale. Biomechanical properties of the right tibia were determined by a three-point bending test in an
Instron Universal Testing Machine (USA, 4444 model, 1 KN load cell). To measure bone mineral content (BMC g/cm2) and
bone mineral density (BMD-g/cm2) of the right femur densitometry (DEXA Hologic Inc Discovery model-Belford, MA, USA)
analysis was carried out by using specific software for small animals. The distribution of all variables was tested for normality by
using Shapiro Wilks W test. Data was analyzed by Kruskal Wallis One-Way Analysis of Variance on Ranks followed by the
Student Newman Keulss post-hoc test. STATISTICA version 7.0 (data analysis software system - StatSoft Inc.) was used to
carry out statistics analysis. Values of p
Conclusions:
The lesion procedure caused severe degradation in behavioral performance, as measured by the BBB score. Injured animals did
not present any recovery in their general motor behavior and none of them presented plantar placement of the paw with weight
support during the experimental period. Two weeks after surgery, the injured animals showed a statistically significant decrease
in maximal load compared to control animals (P=0.02). Interestingly, the animals sacrificed 4 and 6 weeks after surgery did not
demonstrate any difference in the biomechanical evaluation when compared to control (P=0.42; P=0.44 respectively). BMD had a
significant difference, and a decrease 2 and 4 weeks after SCI (P<0.001).
Keywords: spinal cord injury, bone , osteoporosis
Financial Support: CnPQ and Fapesp
Resumo:21-132
ACUTE RESISTANCE EXERCISE INCREASE THE EXPRESSION OF SYNAPSIN I IN HIPPOCAMPAL
FORMATION OF RATS
Fernandes, J. 1; Pena, L. F. P. 1; Cassilhas, R. C. 2; Mello, M. T. D. 2; Venancio, D. P. 2; Scorza, F. A. 3;

Silva, S. G. D. 1; Cavalheiro, E. A. 3; Arida, R. M. 1
1
Departamento de Fisiologia/Universidade Federal de So Paulo, UNIFESP
2
Dept. de Psicobiologia/Universidade Federal de So Paulo, UNIFESP
3
Dept. de Neurologia/Universidade Federal de So Paulo, UNIFESP
Objectives:
Aerobic exercise has been shown to enhance memory and cognition, facilitate functional recovery following brain injury,
promote neurogenesis in the adult brain, and even ameliorate the mental decline associated with aging. However, the influence of
resistance exercise on brain function and neuroplasticity has not been well estabelished. Therefore, the purpose of the present
study was to analyze the impact of one session of resistance exercise on expression of synapsin I phosphoprotein involved in
vesicle clustering, neurotransmitter release and synaptic transmission.
Twelve adult male wistar rats were distributed in two groups: sedentary control group (CTRL=6) and resistance exercise group
(RES=6). Animals from resistance exercise group were submitted to one session of resistance exercise where its climbed a 1.1-m
vertical (80o incline) ladder with incremental weights secured to their tails. The session consisted of 8 climbs with 8-12 dynamic
movements per climb. Exercise increased significantly synapsin I levels (p>0.05) in the hippocampal formation.
Conclusions:
The present result demonstrate that acute resistance exercise promotes changes in a protein marker of synaptic plasticity,
suggesting that some of the neural adaptations by resistance exercise may be observed in early phase of the trainig program.
Keywords: RESISTANCE EXERCISE, SYNAPSIN I, SYNAPTIC PROTEINS, NEUROPLASTICITY, HIPPOCAMPAL
FORMATION
Financial Support: CNPq, CAPES, FAPESP and CInAPCe-FAPESP
Resumo:21-133
DIFFERENTIAL RESPONSE OF SYNAPTIC AND STRUCTURAL PROTEINS IN THE NERVOUS SYSTEM OF
RATS SUBJECTED TO DIFFERENT TYPES OF MOTOR TRAINING
Garcia, P. C. 1; Real, C. C. 2; Britto, L. R. G. 2; Pires, R. S. 1

1
Universidade da Cidade de So Paulo, UNICID
2
Objectives:
A number of studies performed in rodents have revealed cellular and molecular exercise-induced changes in several brain
structures. Motor skill learning is an essential aspect of development, adult life, and recovery after brain lesions. Learning
requires protein synthesis and is likely that some of the proteins synthesized are involved in structural plasticity. Since different
types of exercise may induce distinct changes in different brain areas, it is important to study the plastic responses generated by
the training of complex motor tasks (acrobatic exercise) and compare them with those involved in rhythmic tasks (forced exercise
on a treadmill). The aim of this study was to evaluate the protein expression of synapsin I (SYS), synaptophysin (SYP), MAP2
and neurofilament (NF68) in the striatum and cerebellum of adult rats subjected to forced and acrobatic exercise.
We used adult male Wistar rats, divided into 3 groups based on types of exercise training, namely control-sedentary (n = 15),
forced exercise (FE) (n = 20) and acrobatic exercise (AE) (n = 20). The FE rats were trained on a treadmill with a maximum
speed of 0.6 Km/h, for 40 minutes, 3 times per week for 30 days. In the AE group, the rat had to move through circuits consisting
of various obstacles during the same period of time as the other group. The methods used for the analyses were
immunohistochemistry and Western blotting. Our results show that the present exercise protocols induced specific plastic
changes in both regions studied, which varied depending on the protein studied. Western blot analysis showed that trained
animals, regardless of the type of exercise, showed a higher expression of structural proteins (MAP2 and NF68) than synaptic
proteins (SYS and SYP). The rats of the FE group showed changes only in expression of the structural proteins, exhibiting a
significant increase of MAP2 (p<0.02).
Conclusions:
Acrobatic exercise induced changes in the expression of synaptic and structural proteins mainly in the striatum, which may
underlie part of the learning of complex motor tasks involved in the protocol. On the other hand, forced exercise promoted
changes only in structural proteins in both the cerebellum and the striatum. Taken together, these results suggest that moderate
physical exercise and acrobatic exercise modulate synaptic and structural proteins in motor brain areas, which may play an
important role in the exercise-dependent brain plasticity.

Keywords: motor training, plasticity, structural protein, synaptic proteins
Financial Support: FAPESP, CNPQ
Resumo:21-134
CHRONIC TREATMENT WITH ASCORBIC ACID ENHANCES CORTICAL SPREADING DEPRESSION IN
DEVELOPING WELL NOURISHED AND MALNOURISHED RATS
Alves, E. V. S. ; Guedes, C. K. R. D. M. ; Viana-da-silva, E. ; Guedes, R. C. A.

Depto de Nutrio, UFPE
Objectives:
Ascorbic acid (AA) is an antioxidant that has been suggested as being important for brain development. However, under certain
conditions AA has also been shown to exert biphasical modulating action on excitability-dependent brain phenomena
(Toxicology 249; 35-39, 2008; Neuroscience 128; 721728, 2004). Here we explore the question of whether AA chronic
administration alters cortical excitability as indexed by the cortical spreading depression (CSD).
Well-nourished (W) and malnourished (M) Wistar rats pups received per gavage AA (60mg/kg/d; n=11), or saline (n=11), or no
treatment (nave group; n=10) during postnatal days 7-28. When they became 30-40days old, under anesthesia, CSD was elicited
at 20 min intervals by KCl-stimulation and electrophysiologically recorded during 4h on 2 points of the right parietal cortex.
ANOVA plus Tukey test showed that M rats presented higher (p
Conclusions:
Chronic AA treatment of developing animals facilitates CSD propagation, and this effect is not modified by early malnutrition.
Data are consistent with the hypothesis that, at the dose of 60 mg/k/d, AA can have a prooxidant effect, as previously postulated
(Toxicology 249; 35-39, 2008).
Keywords: CORTICAL SPREADING DEPRESSION, ASCORBIC ACID, WELL NOURISHED, MALNOURISHED, RATS
Financial Support: CAPES, CNPq, FACEPE, INNT-Rede Glial, IBN-Net.
Resumo:21-135
ROLE OF P2X7 RECEPTOR IN NEURODEGENERATION AND MICROGLIOSIS AFTER FOCAL CEREBRAL
ISCHEMIA IN MICE
Amorim, F. E. 1; Navarro-martins, A. 2; Coutinho-silva, R. 3; Cintra, W. M. 1; Mendez-otero, R. 2

1
Laboratrio de Farmacologia Molecular, ICB/UFRJ
2
Laboratrio de Neurobiologia Celular e Molecular, IBCCF/UFRJ
3
Laboratrio de Imunofisiologia, IBCCF/UFRJ
Objectives:
Strokes are second cause of death of world and the first in Brazil. Its shown that the damage caused by ischemia may be
aggravated by secondary events such as the inflammatory response. Its known that the purinergic P2X7 receptor (P2X7R) is able
to activate microglia inducing it to release pro-inflammatory factors. The aim of this study was to investigate the effect of
blocking P2X7R in microglia and neurodegeneration after permanent middle cerebral artery occlusion (MCAo) by
electrocauterization.
C57Bl6 females mice received three doses (45.5 mg/kg - 3 mg/ml, via ip, in PBS containing 0.2% DMSO) of Brilliant Blue G (a
P2X7R antagonist) or vehicle at intervals of 48 hours. The first dose was one day before MCAo. For analysis of lesion volume,
Triphenyl Tetrazolium Chloride (TTC) reaction was made on coronal sections (1 mm thickness) of ischaemic mice 4 days after
MCAo. For immunohistochemical reactions, animals (4 days post-MCAo) were perfused with 4% paraformaldehyde,. Brains
were removed and coronal sections were taken (20 m) in a criostat. To determine lesion area and density of degenerating
neurons we used the Fluoro-Jade C staining, a marker of early neuronal degeneration. Inflammatory infiltrate was analyzed by
double labeling with GSA I-B4 and anti-ED1 (respectively, total and activated microglia). All evaluations were done by t test (p
Conclusions:
There was no difference in rate of microglial activation (activated microglia / microglia total) in both groups, experimental (96.24
0.54) and control (96.07 0.46), P = 0.812. This study demonstrated P2X7R blocking after focal cerebral ischemia by MCAo
in mice decreased neuronal death in ischemic penumbra.
Keywords: Focal Ischemia, Middle Cerebral artery, Oclussion, P2X7, Stroke
Resumo:21-136
OXYTOCINERGIC CONTROL OF FOOD INTAKE IS MODULATED BY ESTROUS CYCLE.
Lima, A. A. 1; Xavier, S. D. O. 1; Feitosa, V. L. C. 1; Reis, F. P. 2; Arago, J. A. 1; Lucca Jr, W1

1
Dept. de Morfologia da Universidade Federal de Sergipe, DMO-UFS
2
Faculdade de Medicina da Universidade Tiradentes, FM-UNIT
Objectives:
To evaluate estrous cycle modulations on the oxytocinergic control of food intake.

After seven days of recover from the cannula implantation surgery, the metestrus and proestrus phases animals were fastened for
48 hours and received at 7:00 h a microinjection (5 uL) of OT (1 g/L - Novartis) in the lateral ventricle. Immediately after the
infusions, it was offered access to balanced ration, and the amount of ingested food was measured during 90 minutes after the
beginning of the food access. Following, the animals were anesthetized with an intraperitoneal injection (1 mL / 100 g of body
weight) of ketamine (80 mg/kg) and xylazine (8 mg/kg) and 30 mL of 0.01M sodium phosphate buffer (pH 7.4) were perfused
through the ascending aorta followed by 300 ml of 4% paraformaldehyde in 0.1 M sodium phosphate buffer (pH 7.4).The other
were sacrificed and wasted, according to the protocol. It was observed that 11% of the animals submitted to the cannula
implantation surgery, altered the cycle regularity and remained in metestrus phase, presenting sometimes a diestrus phase.17 of
31 rats submitted to cannula implantation in the lateral ventricle received isotonic saline microinjection (5 uL) and 14 received
OT microinjection (5 uL). After 48 hours of fast, 9 rats subjected isotonic saline icv microinjection presented food intake of 2,25
( 0,1897 ) g/100g body weight during the 90 minutes that ration was available. The 14 rats subjected OT icv microinjection
presented food intake of 1,45 ( 0,1727) g/100g body weight during the 90 minutes that ration was available. 9 of 16 rats in
metaestrus phase submitted to cannula implantation in the lateral ventricle received isotonic saline microinjection (5 uL) and 7
received OT microinjection (5 uL). After 48 hours of fast, 9 rats subjected isotonic saline icv microinjection presented food
intake of 2.07 ( 0.2536) g/100g body weight during the 90 minutes that ration was available. The 7 rats subjected OT icv
microinjection presented food intake of 1,22 ( 0,1562) g/100g body weight during the 90 minutes that ration was available. 8 of
15 rats in proestrus phase submitted to cannula implantation in the lateral ventricle received isotonic saline microinjection (5 uL)
and 7 received OT microinjection (5 uL). After 48 hours of fast, 8 rats subjected isotonic saline icv microinjection presented food
intake of de 2,46 ( 0,2849) g/100g body weight during the 90 minutes that ration was available. The 7 rats subjected OT icv
microinjection presented food intake of 1,67 ( 0,2957) g/100g body weight during the 90 minutes that ration was available. The
difference between groups wasnt considered statistically significant at p <0.05.
Conclusions:
The OT centrally administered lowers the ration intake of Wistar female previously submitted to a fast and the gonadal steroids
modulate this oxytocinergic inhibitory action in the food intake control.
Keywords: Ocitocina, Ingesto, Esterides gonadais
Financial Support: FAPITEC
Resumo:21-137
ACTIVATION OF C-JUN N-TERMINAL KINASE (JNK) DURING MITOSIS IN RETINAL PROGENITOR CELLS.
Ribas, V. T. ; Gonalves, B. S. ; Linden, R. ; Chiarini, L. B.

Instituto de Biofsica Carlos Chagas Filho, IBCCF/UFRJ
Objectives:
During development of rat retina, cells are found in various stages of development: proliferation, differentiation and programmed
cell death. In this study, we investigated the behavior of c-Jun N-termianl Kinase (JNK) and two of its targets, ATF-2 and c-Jun,
during mitosis of progenitor cells in newborn rat retina.
Retinal explants from newborn rats were kept in vitro for various intervals and under distinct treatments. Sections of retinal
explants were used to detect of JNK, ATF-2 and c-Jun phosphorylation by immunohistochemistry, and examined through both
fluorescence and confocal microscopy. Mitotic cells were identified by chromatin morphology, histone-H3 phosphorylation, and
localization in the retinal tissue. The subcellular localization of phosphorylated proteins was analyzed by double staining with
both DAPI and an antibody to each phosphorylated protein. Phosphorylation of JNK was also examined by Western blot. The
results showed that in the retina of newborn rats (P1) JNK is phosphorylated during mitosis of progenitor cells, mainly during the
early stages of mitosis. It was shown that JNK1/2, but not JNK3, are phosphorylated in mitotic cells. The transcription factor
ATF-2 is not phosphorylated in mitotic cells, whereas transcription factor c-Jun is phosphorylated on both residues serine 63 and
serine 73 at all stages of mitosis. Phosphorylated c-Jun was detected also in the centrosome in mitotic cells. Inhibition of JNK
induced cell cycle arrest, specifically in mitosis. Treatment with the JNK inhibitor decreased the number of cells in anaphase, but
did not alter the number of cells in either prophase/prometaphase or metaphase. Moreover, cells with aberrant chromatin
morphology were found after treatment with the JNK inhibitor. Inhibition of JNK, p38, PKC, ERK or PI-3K did not block the
phosphorylation of c-Jun. Finally, inhibition of JNK induced programmed cell death in progenitor cells in newborn rat retina.
Conclusions:
These data indicate that JNK may play an important role during mitosis of progenitor cells in newborn rat retina.
Keywords: c-Jun N-terminal Kinase, Mitosis, Retina
Financial Support: Capes, CNPq, FAPERJ, PRONEX-RJ.
Resumo:21-138
EFFECTS OF ACETYLCHOLINE INJECTED INTO THE NUCLEUS OF THE SOLITARY TRACT ON
SYMPATHETIC AND PHRENIC NERVE ACTIVITIES
Furuya, W. I. 1; Zoccal, D. B. 2; Menani, J. V. 1; Colombari, E. 1; Colombari, D. S. A. 1

1
Universidade Estadual Paulista - Faculdade de Odontologia, UNESP
2
Universidade Federal de Santa Catarina, UFSC
Objectives:
A cholinergic system was identified within the nucleus of the solitary tract (NTS) of the rat. Accordingly, it has been shown that
acetylcholine (ACH) microinjected into the nucleus of the solitary tract (NTS) induces hypotension and bradycardia (da Silva, L.
G. et al, 2008, AJP, 295, p R1774). However, the effects of ACH in the NTS on sympathetic nerve activity and phrenic discharge
have not been evaluated yet. Thus, the aim of this study was to test the effects of ACH microinjected into the medial NTS
(mNTS) on sympathetic and phrenic nerve activity.
Decorticated arterially-perfused in situ preparations were made from male Holtzman rats (60-80 g, n=5) to record the activities of
thoracic sympathetic (tSNA) and phrenic nerves (PNA). In these preparations, microinjections of saline (60 nl) and ACH (0.1, 1.0
and 10 nmol/60 nl) were performed randomly into the mNTS and the alterations in tSNA and PNA were evaluated. It was
observed that increasing doses of ACH (0.1, 1.0 and 10 nmol/60 nl) into the mNTS significantly inhibited SNA, whose peak of
response was comparable among the doses (respectively, -51 5.7%, -64 4.2%, -60 1.4%, vs. saline: -8 8% of baseline, p <
0.05). However, the decreases in SNA measured over 30 s after ACH were dose-dependent (-11 5.3%, -22 5.8%, -60 1.4%,
vs. saline: 0 4% of baseline, p < 0.05). Conversely, ACH microinjections into the mNTS did not produce significant reductions
in PNA frequency (-3.5 1.7, -5.2 0.9, -7.3 1.3, vs. saline: -1.4 1.8 bursts per minute, p > 0.05).
Conclusions:
These data suggested that cholinergic system at the mNTS level is involved in the regulation of SNA, possibly acting on neurons
involved in the baroreflex pathways. On the other hand, ACH in the mNTS, at least in the doses studied, may not influence the
generation of PNA.
Keywords: acetylcholine, nucleus of the solitary tract, phrenic nerve activity, sympathetic nerve activity
Resumo:21-139
CIGARETTE SMOKE AND/OR ETHANOL DURING ADOLESCENCE OF MICE: MEMORY AND LEARNING
EFFECTS DURING EXPOSURE AND WITHDRAWAL.
Graa, A. C. C. ; Naiff, V. F. ; Carvalho, A. R. ; Manhes, A. C. ; Filgueiras, C. C. ; Villaa, Y. A.

Departamento de Cincias Fisiolgicas/ UERJ, IBRAG/UERJ
Objectives:
AIM: Adolescents often associate tobacco smoking and consumption of alcoholic beverages. In spite of this frequent association,
little is known about the basic neurobiology of the dual exposure in the adolescent brain. This study sought to evaluate the effect
of tobacco smoke, ethanol and the double exposure tobacco smoke+ethanol on memory/learning of mice during exposure and
after a short period of withdrawal.
METHODS: Four groups of Swiss mice were used to assess the impact of cigarette smoke and/or ethanol consumption on
memory/learning: 1) SMK+ETOH - animals exposed to smoke generated from burning research cigarettes (Kentucky University,
Louisville, KY, USA) containing 0.73mg of nicotine in a cigarette smoking machine (Teague Enterprises, Davis, CA) and i.p.
injection of ethanol solution (2g/kg, 25%); 2) SMK - cigarette smoke and i.p. saline; 3) ETOH - air and i.p. ethanol; 4) VEH - air
and i.p. ethanol. Smoke exposure was carried out from postnatal day (P) 30 to P45 for 8h per day in order to mimic the pattern of
consumption of regular smokers that discontinue consumption during sleep. Ethanol exposure was carried out every other day to
mimic binge consumption. At P44, animal behavior associated with memory/learning was evaluated in the step down passive
avoidance test (SD). Animals were placed on a platform and were allowed 3 min to descend from it, at which time they received
a foot shock (0.2 mA, 3 s). Three and 24h after the initial testing (AIT), animals were retested in the SD (shock was not
administered). The latency to leave the platform was the measured variable. Data were analyzed using ANOVAs. RESULTS: Our
results showed a treatment effect on P45 (P=0.007) 3 h AIT. This can be explained by the fact that SMK, SMK+ETOH e ETOH
animals presented a shorter latency to leave the platform when compared to VEH animals (Pairwise comparisons: ETOH and
VEH: P=0.009; SMK and VEH: P=0.024; SMK+ETOH and VEH: P=0.007). Twenty-four h AIT, we found a treatment effect
(P=0.028) as well as a sex effect (P=0.055), which can be explained by the fact that female SMK e SMK+ETOH presented
shorter latency to leave the platform when compared to VEH females (SMK and VEH: P=0.049; SMK+ETOH and VEH
P=0.027). At P50, three h AIT we did not obtain significant differences between groups. Twenty-four hours AIT, we observed an
interaction between treatment and sex (P=0.045) showing that male ETOH e SMK+ETOH presented shorter latency when
compared to male VEH (ETOH and VEH: P=0.004; SMK+ETOH and VEH: P=0.002).
Conclusions:
CONCLUSION: Exposure tobacco smoke, ethanol or the combined exposure during mice adolescence affects memory
performance in the step down passive avoidance test.
Keywords: Cigarette smoke , Ethanol , Memory and learning , Mice, Adolescence
Financial Support: FAPERJ, CNPq, CAPES and SR2-UERJ
Resumo:21-140
DYNAMICS OF SPONTANEOUS AND REFLEX BLINKS IN THE BURROWING OWL (ATHENE CUNICULARIA).
Vieira, P. G. 1; Almeida, F. 1; Souza, J. K. S. 2; Pinto, M. A. S. 2; Sousa, J. P. M. D. 2; Tierra-criollo, C. J. 2;

Baron, J. 1,2
1
Fisiologia e Biofsica / Instituto de Cincias Biolgicas, UFMG
2
Engenharia Eltrica / Escola de Engenharia, UFMG
Objectives:
Several times a minute, eyeblinks produce transient visual disruptions that are usually not registered consciously. As a
preliminary step towards understanding the central mechanisms underlying the perceptual filling-in of such temporal scotomas,
we presently report kinematics data of spontaneously occurring and air-puff evoked blinks in the animal species that we selected
as a model for addressing this issue, namely the burrowing owl (Athene cunicularia).
Methods: We recorded 78 spontaneous and 82 induced blinks in two head-restrained, non-anesthetized owls using a computerassisted infrared pupillometer developed by our group. Eyelid and nictitating membrane movements were measured at a
sampling rate of 120 Hz. To induce a blink reflex, we used a customized air-puff device that comprised an electronically
controlled valve unit, an air reservoir and a flexible thin plastic tube for funneling the air to the cornea and periorbital region.
The air-puff pulse duration was set to 20 ms. For data analysis, we used our own Labview-based pupillometric software, Matlab
(MathWorks, Natick, MA, USA) and the public domain image processing program ImageJ 1.44 (NIH,
http://rsb.info.nih.gov/ij/).Population measurements were presented as median, using the 25th and 75th percentile as an
indicative of population dispersion. All experimental and animal care procedures were approved by the University Ethics
Committee for Animal Experimentation (CETEA, license n 106/10). Results: Spontaneous blinks were characterized by a lack of
complete eyelid closure, occluding only 30% (16.6 38.2%) of the pupil. In contrast, the nictitating membrane almost always
swept the entire eye globe. Duration of spontaneous blinks was 350 ms (275 450 ms) with a down phase duration of 125 ms
(108 150 ms) and up phase of 208 ms (167 300 ms). Evoked blinks resulted in almost complete covering of the eye globe.
Duration of reflex blinks was 575 ms (460 756 ms) with a down phase duration of 158 ms (125 217 ms) and up phase of 408
ms (317 556 ms). Latency of eyelid response to air puffs was short (92 ms). For both spontaneous and reflex blinks, the down
phase of the nictitating membrane occurred concomitantly with that of the eyelid; the upward phase of the nictitating membrane
was nevertheless much slower.
Conclusions:
Our study reveals that the down phase is fairly constant for both types of blinks, whereas the up phase differs markedly in
duration. At least three factors may explain this finding: (1) differences in palpebral response amplitude increasing the traveling
distance for fully reopening the eye; (2) variation in the animal central state due to corneal e periorbital stimulation as opposed
to absence of stimulation; (3) involvement of different neuronal components involved in mediating the two types of blink. Which
of these factors explain our data remains to be determined.
Keywords: Blink dynamic, owl, temporal scotomas
Financial Support: FAPEMIG, CNPq
Resumo:21-141
AV3V LESION DOES NOT CHANGE THE CARDIOVASCULAR RESPONSES TO CARBACHOL IN THE NTS.
Zanella, D. C. ; Menani, J. V. ; Paula, P. M. D.

Universidade Estadual Paulista "Jlio de Mesquita Filho", Unesp
Objectives:
Studies from our laboratory have shown that lesions of the preoptic periventricular tissue surrounding the anteroventral third
ventricle (AV3V region) reduced the pressor responses to glutamate and adenosine 5'-triphosphate (ATP) into the nucleus of the
solitary tract (NTS). This lesion also reduced the hypotension caused by alpha, beta-methyl ATP (a selective P2X purinergic
receptors agonist) into the NTS. Similar to alpha, beta-methyl ATP, carbachol (cholinergic agonist) when injected into the NTS
in awake rats produced a drop in mean arterial pressure (MAP) and heart rate (HR). Therefore, in the present study, we
investigated the effects of acute (1 day) electrolytic lesions of the AV3V region on the hypotension and bradycardic responses
induced by injections of carbachol into the NTS in unanesthetized rats.
Male Holtzman rats (n=6/group) with sham or electrolytic (2 mA x 10 s) AV3V lesions and a stainless steel cannula implanted
into the NTS were used. The hypotension and bradycardic responses produced by unilateral injections of carbachol (2.5 nmol/100
nl) into the NTS (-25 1 mmHg and -154 8 bpm) did not change in 1 day AV3V-lesioned rats (-29 3 mmHg and -134 7
bpm, p>0.05).
Conclusions:
The results show that the integrity of the AV3V region is not important for the cardiovascular responses produced by cholinergic
activation into the NTS but is essential for the cardiovascular responses by purinergic and glutamatergic activation into the NTS.
Keywords: arterial pressure, AV3V region, carbachol , glutamatergic , NTS
Financial Support: CNPq and FAPESP
Resumo:21-142
IMPAIRED INSULIN SIGNALING IN A PRIMATE MODEL OF ALZHEIMER`S DISEASE: LINK WITH TYPE-2
DIABETES
Forny-germano, L. 1; Bomfim, T. R. 1; Lyra E Silva, N. M. 1; Christophe-houzel, J. 1; Brito-moreira, J. 1;

Klein, W. L. 3; Munoz, D. P. 4; Ferreira, S. T. 1; de Felice, F. G. 1
1
3
4
Northwestern University, NU
Queen's University, queensu
Objectives:
Alzheimers disease (AD) has been linked to defective brain insulin signaling, a proposed third type of diabetes (Curr.
Alzheimer Res. 4:147, 2007; BMB. Rep. 42:475, 2009; FASEB J. 22:246, 2008). Although this intriguing connection between
AD and diabetes has been suggested, a major unknown is the mechanism by which insulin resistance develops in AD brains. In
type 2 diabetes, tumor necrosis factor- (TNF-) signaling stimulates c-Jun N-Terminal Kinase (JNK). This results in serine
phosphosphorylation of the insulin receptor substrate (IRS-1), blocking downstream signaling and triggering insulin resistance
(Science 271:665, 1996). One link between diabetes and AD was found in the ability of A oligomers - toxins that accumulate
in AD brains and instigate synaptic damage - to activate the JNK/TNF- pathway, leading to phosphorylation of IRS1pSer636. Currently, there is no effective treatment for AD, and the pursuit of novel disease-modifying therapeutics is the object
of intense investigation. Although a few therapeutic drugs (JAMA 304:1903, 2010) and antibodies targeting A oligomers
(Neurobiol. Aging 30:1728, 2009) have shown some promises in preclinical studies and early clinical trials, unfortunately a
number of trials have failed to show any efficiency in AD patients (JAMA 304:1903, 2010). Part of this discrepancy lies in the
difficulty to translate therapeutic approaches developed in rodent models to the specific conditions of the human disease. In an
attempt to get closer to the complexity of primate brains, here we extend our model of chronic intracerebroventricular injection of
A oligomers in rodents to macaque monkeys and investigate if Abeta oligomers are capable of triggering diabetes-like effects
on the JNK/TNF- pathway in primates.
Freshly prepared A oligomers were injected in the cerebral ventriculs of three adult (~9 years of age) cynomolgus monkeys
(Macaca fascicularis, weight 4.7-7.0Kg). Monkeys received 100g of A 3 times a week for 3 weeks. One sham-canulated
monkey served as control. Brains were removed and later, immunohistochemistry for pJNK and IRS-1pSer636 was performed on
40 m thick frozen cut sections. Analysis revealed a marked activation of JNK in the hippocampus in A oligomers injected
monkeys, when compared to control. In addition, pixels per area quantification using NIH Image Java indicated that levels of
IRS-1pSer636 were significantly increased in the hippocampus (p=0.0002) and in the temporal cortex (p
Conclusions:
Our results reinforce the link between diabetes and AD. Furthermore, considering the dearth of animal models that truly
recapitulates the main features of Alzheimers disease, this new non-human primate model offers a potential to provide
insights into central aspects of AD that may be exclusively present in primates.
Keywords: Alzheimer`s disease, c-Jun N-Terminal Kinase, Insulin receptor substrate, Primate model, Type-2 Diabetes
Financial Support: CNPq, FAPERJ, INNT.
Resumo:21-143
PUPILLARY LIGHT REFLEX IN THE BURROWING OWL (ATHENE CUNICULARIA).
Souza, J. K. S. 1; Vieira, P. G. 2; Tierra-criollo, C. J. 1; Baron, J. 1,2

1
Engenharia Eltrica / Escola de Engenharia , UFMG
2
Objectives:
The pupillary light reflex (PLR) refers to the constriction of the pupil produced by an increase in retinal illumination. Here, we
provide PLR data obtained from adult burrowing owls (Athene cunicularia) as a means to gain further insights into the functional
properties of the neural pathway mediating this reflex in birds.
Methods: PLR was measured in two head-restrained, non-anesthetized owls using a computer-assisted infrared pupillometer
developed by our group. Pupil diameter was measured at a sampling rate of 120 Hz with an estimated spatial resolution of 0.07
mm. Visual stimuli were presented either mono- or binocularly from a 19-inch CRT monitor placed 35 cm from the eyes of the
animal and consisted of brief flashes (100 ms) of large uniform fields (angular subtense: 39.4 x 26.6 degrees) of achromatic light
that varied randomly in luminance (steps: 15, range: 15 to 162 cd/m2). All recording sessions were initiated after 20 minutes of
dark adaptation. Data were analyzed using our own Labview-based pupillometric software, Matlab (MathWorks, Natick, MA,
USA) and the public domain image processing program ImageJ 1.44 (NIH,http://rsb.info.nih.gov/ij/). Results are presented as
arithmetic means SD. All experimental and animal care procedures were approved by the University Ethics Committee for
Animal Experimentation (CETEA, license n 106/10). Results: Mean pupil diameter varied from about 10 mm under near-dark
conditions to just over 5 mm (4-fold reduction in retinal illuminance level). Saturation in pupillary response was observed for
stimuli above 65 cd/m2. Response onset latency was fairly constant across all luminance levels (mean value: 54.07.6 ms). At the
highest luminance tested, time to reach peak constriction was 136.810.0 ms, yielding in a constriction speed of 58.64.2 mm/s;
return to baseline level (post-stimulus dilatation) was significantly slower (duration: 813.2267.4 ms; speed: 20.92.0 mm/s). At
the lowest luminance, time to reach peak constriction was 85.19.3 ms, resulting in a constriction speed of 12.64.1 mm/s; return
to baseline level (post-stimulus dilatation) was also significantly slower (duration: 725.0310.2 ms; speed: 10.13.1 mm/s).
There was no statistical difference in pupillary response between monocular and binocular viewing conditions.
Conclusions:
Our study reveals that PLR is about six times faster in burrowing owls than in primates. Another interesting difference is that, in
the owl, response latency does not vary as function of luminance and no consensual response is observed. Whether such
differences are solely explained by the striated musculature of the owls iris (as opposed to smooth muscles in mammals) remains
an open question.
Keywords: Pupillary light reflex, owl, pupillometry
Financial Support: FAPEMIG, CNPq, CAPES
Resumo:21-144
HUMAN DIFFERENCE LIMENS FOR FLICKER FREQUENCY OF ACHROMATIC LIGHT
Akemy, B. 1; Sousa, J. P. M. 2; Pinto, M. A. D. S. 2; Tierra-criollo, C. J. 2; Baron, J. 1,2

1
2
Programa Ps-Graduao Engenharia Eltrica/Escola Engenharia, UFMG
Objectives:
The temporal visual sensitivity of human perception may be assessed in a number of ways. Here, we aim at estimating differential
thresholds for detecting slight differences in flickering frequency increments and decrements.
Experimental sessions took place in a soundproof and lightcontrolled booth. Subjects (n=14) were accommodated in an
armchair positioned 57 cm away from the stimulation source, provided by flickering white LEDs driven by a portable,
microcontrolled stimulator developed by our group (J.Neurosci. Methods, 197, 8291; 2011). Perceptual thresholds were
measured using a temporal twoalternative forced choice (2AFC) procedure in combination with a method of constant stimuli.
Each trial consisted in presenting a reference stimulus flickering at 10 Hz (F1) during 2s, followed by a test stimulus (F2)
flickering at a slightly different frequency (n=14; incrementsdecrements: 0.2 Hz) or at the same frequency as F1 (n=4) during
the same period. An interval of 4s was left between F1 and F2. Using a response button, subjects were asked to report which
stimulus in the trial sequence had the highest flickering frequency (one press: F1; two presses: F2). No feedback about the
accuracy of the judgments was given. All procedures adopted in this study have been approved by the University Human
Research Ethics Committee (COEP, ETIC 46708). All subjects readily perceived flickering frequency increments and
decrements above 2 Hz. The average hit rate when F2 was higher than F1 was 86,96% (SD: 9,33%) and 76,82% (SD: 9,24%)
when the opposite stimulation condition occurred. Threshold estimates across subjects were fairly constant and approximated 0.9
Hz and 0.4 Hz for frequency decrements and increments, respectively. For most subjects, a robust subject criterion bias was
however observed, which induced higher hit scores for positive increments. Catch trial analysis revealed that almost all subjects
had a tendency to choose F2 higher than F1 in 23 of the cases.
Conclusions:
Our results are globally coherent with earlier studies that have assessed difference limens for flicker frequency, but using a spatial
2AFC experimental design. Future work will include the testing of other F1 values and the effect of increasing the
inter&minusstimulus interval as a means to assess the retention capability of temporal attributes of basic visual stimuli.
Keywords: temporal visual sensitivity, flickering frequency, estimating thresholds
Financial Support: Brazilian agencies CAPES and FAPEMIG
Resumo:21-145
NUCLEAR FACTOR KAPPA B AS A DUALISTIC MEDIATOR OF CEREBELLAR GRANULE CELL CULTURE
DAMAGE
Franco, D. G. ; Markus, R. P.
Dep. Fisiologia - Inst. de Biocincias, IBUSP
Objectives:
In the central nervous system nuclear factor kappa B (NFKB), a transcription factor primarily related to inflammatory responses,
plays an important role in the modulation of plasticity, development and cell survival (Cell & Mol Immuno. 1:425, 2004). As an
example, in cerebellar granule cell culture (CGC), which has a basal NFKB activation, tumor necrosis factor (TNF) increases
NFKB nuclear translocation leading to an inverted-U shaped dose survival curve. Therefore, very high or low concentrations of
TNF result in cells death, while intermediary concentrations are necessary for cell survival(Bioch et Bioph Acta. 1745:287,
2005). Melatonin, the pineal gland hormone, acts as a cytoprotector and anti-oxidant agent impairing the activation of NFKB in
macrophages (FASEB J. 12:685, 1998) and endothelial cells culture (J. Pineal Res. 46:268, 2009). We have previously shown
that in CGC, melatonin inhibits lipopolysaccharide (LPS)-induced NFKB translocation, but has no effect on basal NFKB nuclear
content (Annals XV Congress SBBC 2010, pp 130). This fine tune balance a critical factor involved in cell survival lead us to
investigate the NFKB manipulation effects on cell damage. Therefore, we evaluated cytotoxicity promoted by the inhibition of
basal NFKB by PDTC (pyrrolidine dithiocarbamate), a competitive blocker NFKB dimers binding to DNA kappa B responsive
element, or the NFKB activated by LPS in the presence or absence of melatonin in CGC.
Granule cells isolated from rat Wistar (7-8 days-old) cerebellum were cultivated for 8 days in DMEM (10% FBS). At the 7th/8th
day, cells were incubated with LPS (0.1 and 1g/mL) in the presence or absence of melatonin (0.001, 0.1 and 1g/mL) or PDTC
(0.001 to 10g/mL) for 12h. Cellular membrane damage was determined by lactate dehydrogenase (LDH)-leakage into the
medium. Statistical analyses were performed using ANOVA followed by the NewmanKeuls test. Values of p
Conclusions:
In CGC, the balance of NFKB activation is an important factor for cell survival. As we know, melatonin has no effect on basal
nuclear NFKB, so that, it had no effect on membrane damage. In the other hand, PDTC that promotes inhibition of nuclear NFKB
translocation promoted an increase in cytotoxicity. The nuclear translocation of NFKB by LPS did not result in cell toxicity,
however, the concomitant incubation of LPS and melatonin increased cell damage. Before considering that melatonin is not
exerting its cytoprotective effect, it is important to point out that lower concentrations were more deleterious than higher
concentrations. A special and inverse related dose-response regarding the relationship between cell survival and NFKB activity
was also observed with PDTC, since the lower concentrations were also more deleterious than the higher ones. Therefore, this
work shows a new perspective for evaluating NFKB as a pivotal transcription factor involved in the balance between the classical
pro and anti-inflammatory responses in cerebellar neurons.
Keywords: cell damage, cytoprotection, granule cell culture, melatonin, nuclear factor kappa B
Financial Support: FAPESP, CNPq, CAPES
Resumo:21-146
ACUTE DEXAMETHASONE IMPROVES PERFORMANCE AND SEXUAL MOTIVATION IN MALE RATS
Teixeira, A. S. ; Santiago, M. B. ; Giusti-paiva, A.

Instituto de Cincias Biomdicas/Univ. Federal de Alfenas, ICB/ UNIFAL-MG
Objectives:
The study of sexual behavior in rats involves the analysis of motivation and sexual performance. The distinction is between
seeking sexual contact and be able to perform the copula. This behavior is necessary for the preservation of species and not for
individual survival. Thus, it differs considerably from other spontaneous behaviors such as feeding, hydration, respiration or
thermoregulation. Studies show that stress and consequent activation of the hypothalamic-pituitary-adrenal (HPA) axis and
secretion of glucocorticoids can alter various hormonal and behavioral parameters. The aim of this study was to assess sexual
motivation and performance of male rats and its possible modifications before of acute administration of dexamethasone (DEX),
a synthetic glucocorticoid.
Methods:Wistar rats were housed in groups of four animals per cage in a room maintained under a light-dark cycle of 12h at 21
1C. In experiment 1, adult male rats (350-450g) sexually experienced were previous treated with vehicle (control, n = 8) or DEX
(1 mg/kg, n = 10) 2 hours before of exposed to receptive females (220-280g) by 40 minutes. Later the tapes were analyzed and
quantified the following parameters: mount latency (ML), intromission latency (IL), number of incomplete mounts (IM), number
of intromissions (NI), ejaculatory latency (EL); latency to first mount after ejaculation (LFMA), latency to first intromission after
ejaculation (LFIA), number of incomplete mounts after ejaculation (IMA), number of intromissions after ejaculation (NIA), total
number of ejaculations (NE). In experiment 2, male rats (290-350g), sexually inexperienced, were treated with vehicle (control, n
= 9) or DEX (1mg/kg, n = 11) and, 2 hours after, were placed in an apparatus where there was a receptive female (sexual
incentive) and a sexually experienced male (social incentive), by 20 minutes. After analyzing the videos were quantified the
following parameters: time in seconds spent in the female incentive zone (FIZ) and male incentive zone (MIZ), frequency of
visits to each incentive zone. Then we calculated the score of preference. The behaviors of the male rats were started in the dark
cycle, from 20:00 pm. The experiments were conducted with the approval of the Ethics Committee of the Federal University of
Alfenas (protocol 306/2010). Results With respect to experiment 1, we observed that administration of DEX reduces the NI (14.8
1.7 vs. Control: 24.1 3.6 intromission, p = 0.02), reduction in EL (671 130 vs. Control: 1155 151 seconds, p = 0.02),
reduced LFMA (252 13.5 vs. Control: 327 19.2 seconds, p = 0.004), reduction in LFIA (253 13 vs. Control: 333 19.9
seconds, p = 0.003) and increase in NE (3.8 0.4 vs. Control: 2.3 0.2 ejaculations, p = 0.01). Regarding the ML and IL, which
were not affected by DEX administration since these parameters are influenced by sexual experience and it was common to both
groups. Considering the experiment 2, we observed that administration of DEX increased the FIZ (738.5 42.88 vs. Control:
584.4 45.74 seconds, p < 0.05), reduced the MIZ (219.1 26.58 vs. Control: 333.6 48.18 seconds; p < 0.05), increased the
score of preference for female (0.77 0.03 vs. control: 0.64 0.04, p < 0.05). The other parameters showed no significant
difference.
Conclusions:
The behaviors changes caused by the acute administration of DEX suggest a better performance and sexual motivation in male
rats.
Keywords: Dexamethasone, Glucocorticoids, Hormonal control of behavior, Male rats, Sexual behavior
Financial Support: Fapemig; CNPq; CAPES; FINEP
Resumo:21-147
FUNCTIONAL CORRELATES OF ABERRANT BUNDLES IN CALLOSAL DYSGENESIS
Lazarev, V. V. 1; Monteiro, M. C. 1,2,3; Oliveira-souza, R. 2; Deazevedo, L. C. 1; Moll, J. 2; Lent, R. 3;

Tovar-moll, F. 2,3
1
Instituto Fernandes Figueira, IFF
2
Instituto D'Or de Pesquisa e Ensino , IDOR
3
Objectives:
Callosal dysgenesis (CD) is characterized by a developmental failure of formation of the major commissural bundle. Previous
studies had demonstrated structural evidence for abnormal white-matter pathways (Probst and Sigmoid bundles) in this
pathology. However, evidence for a functional role for the Sigmoid bundle is still lacking. Here we combined diffusion tensor
imaging (DTI) with quantitative electroencephalography (EEG) in order to address, in vivo, the functional correlates of these
pathways and the relationship between structural and functional neuroplasticity in human CD.
Volumetric anatomical (3DT1) and DTI (2,5mm isotropic voxel) images were acquired (3T, Achieva Philips) in five patients
with CD and fifteen healthy volunteers. Neurophysiological assessment was performed by 16-electrode EEG registration (Bio-
logic, USA) and analysis of the Coefficients of Coherence (CChr) (Brainsys - Neurometrics, Russia) in five EEG frequency
bands. The Intelligence Quotient Test was obtained from all the subjects. Group comparison was performed using the t-test. The
patients had lower intelligence quotients within normal range. In the patients, conventional images showed typical anatomical
characteristics of CD, including partial dysgenesis (n=2), agenesis (n=2) or hypoplasia (n=1) of the corpus callosum (CC). In
addition, DTI and tractography confirmed the presence of two abnormal white-matter tracts: (i) the Probst bundles (PB)
connecting intrahemispheric cortical regions in all the patients and, (ii) the Sigmoid bundles, connecting the frontal lobe with the
contralateral occipito-parietal cortex (n=3, except for agenesis). General analysis of CChr showed an increased intrahemispheric
and decreased interhemispheric coherence in patients as compared to controls. In the patients with partial CD or CC hypoplasia, a
relatively higher interhemispheric coherence in the frontal areas was observed. These three patients also presented higher
coherence connectivity between the right frontal and left parietal areas and, in certain frequency bands, bilateral increase in some
other long-distance contralateral coherent connections of the fronto-polar and frontal regions with the occipital, parietal and
central ones.
Conclusions:
The results corroborate previous anatomical findings in CD, showing the presence of abnormal bundles formed during the
development. The neurophysiological results correlate with observed structural changes, since an increased intrahemispheric
connectivity may be related to the presence of the Probst bundle. In addition, the anatomical connectivity of the Sigmoid bundle
may underlie the higher interhemispheric coherence observed between contralateral anterior and posterior areas in partial CD and
CC hypoplasia. The changes in electrophysiological connectivity observed in CD might be the functional counterpart of the
abnormal white-matter connections detected in these patients.
Keywords: callosal dysgenesis, diffusion tensor imaging, EEG Coherence
Financial Support: CAPES, CNPq, FAPERJ
Resumo:21-148
EFFECTS OF 8-PHENYLTHEOPHYLLINE ON CATALEPSY BEHAVIOR INDUCED BY HALOPERIDOL, IN
MICE
Arajo, R. S. ; Ribeiro, T. S. ; Souza, A. S.

Fundao Universidade Federal de Mato Grosso do Sul , UFMS
Objectives:
Adenosine plays an important hole on basal nuclei functions; however dopamine and adenosine interactions on these nuclei are
not completely understood. The aim of this study was assess the effects of an A1 selective adenosine antagonist, 8phenylteophylline, on catalepsy behavior induced by the D2 dopamine antagonist receptor, haloperidol, in mice.
Swiss male mice, 25-35g weight, were initially injected by intraperitoneal route with saline or haloperidol and after 30 minutes
injected by intraventricular route with saline or 8-phenylteophylline (5 and 10nM), performing a total of 6 experimental groups:
(n=8/group): i) saline + saline; ii) saline + 8-phenylteophylline (5nM); iii) saline + 8-phenylteophylline (10nM); iv) haloperidol
(2 mg/kg) + saline; v) haloperidol (2mg/kg) + 8-phenylteophylline (5nM); e vi) haloperidol (2mg/kg) + 8-phenylteophylline
(10nM). After drug administration (5, 35 and 65 minutes) the animals were submitted to catalepsy test measuring the latency
(sec.) on the bar test. The catalepsy data were log transformed (log10 + 1) and analyzed by two-way ANOVA of repeated
measures followed by Duncan post hoc test. The catalepsy time in each experimental group 5, 35 e 65 minutes after drug
administration were: i) saline + saline: 5min=1.130.58s; 35min=1.380.56s; 65min=0.750.25s; ii) saline + 8-phenylteophylline
(5nM): 5min=0.630.26s; 35min=1.130.30s; 65min=1.750.45s; iii) saline + 8-phenylteophylline (10nM): 5min=0.250.16s;

35min=0.380.26s; 65min=0.000.00s; iv) haloperidol (2mg/kg) + saline: 5min=35.753.64s; 35min=115.6330.88s;
65min=177.5032.00s; v) haloperidol (2mg/kg) + 8-phenylteophylline (5nM): 5min=53.1320.05s; 35min=29.139.88s;
65min=35.6315.13s; and vi) haloperidol (2mg/kg) + 8-phenylteophylline (10nM): 5min=104.5036.10s; 35min=68.0036.85s;
65min=41.6320.37s. There was a significant effect of experimental group, no effect of time and significant interaction between
group X time (Two-way ANOVA of repeated measures; group: p<0.05).
Conclusions:
Antagonism of dopamine receptors by haloperidol produced catalepsy on the animals. This effect was partially reverted by 8phenylteophylline an A1 adenosine antagonist receptor in both doses used (5 and 10nM). This effect was observed only 35 e 65
minutes after drugs administration.
Keywords: BEHAVIOR, CATALEPSY, HALOPERIDOL, 8-PHENYLTHEOPHYLLINE , MICE
Financial Support: UFMS, CNPq.
Resumo:21-149
MEDIAN NERVE REPRESENTATION IN RAT S1 AND IMMEDIATE PLASTICITY AFTER ACUTE FOREPAW
DEAFFERENTATION.
Oliveira, J. T. ; Bittencourt-navarrete, R. E. ; Martinez, A. M. B. ; Franca, J. G.

Objectives:
Transection of peripheral nerves interrupts sensory inputs to the central nervous system, resulting in cortical plasticity. In this
study we investigated the representation of the median nerve in rat primary somatosensory cortex (S1) after acute section of all
other brachial plexus nerves.
Three adult Wistar rats had their right brachial plexus transected, except for the median nerve. A contralateral craniotomy was
performed followed by electrophysiological mapping of the representation of the forepaw and adjacent body parts in S1. The
median nerve was then transected and a remapping of the same cortical region was performed. After electrophysiological
mapping small DY crystals were placed at 3 or 4 cortical sites as fiducial markers. The animals were transcardially perfused with
a fixative solution. The cortex was flattened and processed for cytochrome oxidase histochemistry. Superimposition of barrel
fields with the functional maps revealed that all the forepaw barrel subfield (FBS) was responsive to forepaw stimulation carried
by the median nerve alone. Most sites responded to light touch stimuli in the forepaw, fewer sites responded to deep touch or
movement stimulation. After median nerve transection, we observed an expansion of the representation of vibrissae, hindpaw,
and inferior lip into regions previously responsive to the forepaw in the FBS.
Conclusions:
We conclude that the median nerve can relay responses to the whole FBS and that part of this barrel subfield can become acutely
responsive to stimulation of other body parts once the forepaw is completely deafferented.
Keywords: brachial plexus, cortical mapping, median nerve, plasticity, somatosensory cortex
Financial Support: CAPES, Cnpq and Faperj
Resumo:21-150
EFFECT OF CENTRAL INJECTION OF HIDROGEN PEROXIDE ON 2% SUCROSE AND FOOD INTAKE.
Zanella, R. C. ; Menani, J. V. ; Colombari, E. ; Colombari, D. S. A.

Fisiologia e Patologia/ Faculdade de Odontologia Araraquara , UNESP
Objectives:
Reactive oxygen species (ROS) produced endogenously may act in the brain modulating autonomic and behavioral responses.
Previously we have demonstrated that intracerebroventricular (icv) injection of a ROS (hydrogen peroxide, H2O2) reduced the
dipsogenic responses to icv injection of angiotensin II and carbachol (cholinergic agonist) or to intragastric load of hypertonic
saline. In order to test whether central H2O2 effects would be specific to water intake, we investigated the effects of icv injection
of H2O2 on 2% sucrose and food intake.
Male Holtzman rats (280-320 g) with stainless steel guide-cannulas implanted into the lateral ventricle (LV) were used. In the
first protocol (n = 5), animals were trained to ingest 2% sucrose for 2 h daily over 7 days. Phosphate buffered saline (PBS; 1 l)
or H2O2 (5 mol/1 l) was injected into the LV 1 minute before starting the measurement of 2% sucrose intake. In the second
protocol (n = 11), PBS (1 l) or H2O2 (5 mol/1 l) was injected into the LV 1 minute before starting the measurement of food
intake by 24 h food-deprived rats. The treatment with icv H2O2 reduced 2% sucrose intake (0.7 0.4 ml, vs. PBS: 5.1 1.6 ml/2
h, p < 0.05). The treatment with icv H2O2 did not reduce food intake (10.3 0.6 vs. PBS: 10.7 0.9 g/2 h, p >0.05), but reduced
meal-associated water intake (8.3 0.8 vs. PBS: 12.8 1.2 ml/2 h, p < 0.05).
Conclusions:
The results suggest an inhibitory role for H2O2 acting centrally on fluid intake (water and sucrose), without changing food
intake, which excludes a non-specific effect of H2O2 acting centrally on all ingestive behaviors.
Keywords: CENTRAL HIDROGEN PEROXIDE, SUCROSE INTAKE, FOOD INTAKE
Financial Support: CNPq, FAPESP.
Resumo:21-151
OXIDATIVE STRESS IN THE BRAIN OF 21 MONTHS OLD RATS AFTER AGUTE ADMINISTRATION OF AN
AQUEOUS EXTRACT OF THE MEDICINAL MUSHROOM AGARICUS BRASILIENSIS (A. BLAZEI)
S-nakanishi, A. B. de ; Soares, A. A. ; Comar, J. F. ; Natali, M. R. M. ; Peralta, R. M. ; Bracht, A.
Departamento de Bioqumica, Universidade Estadual de Maring, UEM
Objectives:
Reactive oxygen species (ROS) are involved in the mechanism of several neurodegenerative processes related to ageing. The
mushroom Agaricus brasiliensis (formerly known as A. blazei), popularly known as cogumelo do sol, has been amply utilized in
popular medicine as a potential therapeutic agent for treatment of several diseases. It is generally accepted that A. brasiliensis has
antioxidant properties, which are attributed to its contents in vitamins A and C, &beta-carotenes and several phenolic compounds.
If the mushroom also exerts antioxidant effects in the ageing brain, however, is still an open question.
Male Wistar rats, (with 21 months) were treated, by gavage, during 21 days with a lyophillized aqueous extract of A. blazei (200
mg kg-1 day-1). Control rats (same age, aged rats) were treated with saline during the same period. After fasting for 24 hours, the
rats were decapitated, the brain removed and rapidly freeze-clamped in liquid nitrogen. The tissue was homogenized in 100 mM
phosphate buffer (pH 7.4) and used for determining the following oxidative state markers: substances reactive to thiobarbituric
acid (TBARS), reduced glutathione (GSH), reduced protein sulphidryl groups (reduced thiols) and the activities of the enzymes
catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR). Brains of rats that were 3 months old were also
processed. These rats comprise the adult controls. The results revealed that the TBARS levels increased with ageing: 1.590.09
(n=5) and 2.070.17 (n=5) nmol.(mg protein)-1 (p = 0.04) in adults and aged rats, respectively. However, the A. brasiliensis
treatment significantly reduced TBARS of aged rats to 1.550.04 (n=4) (p=0.04 versus aged controls). The GSH and thiol levels
were not changed upon ageing: 2.610.15 (n=5) and 2.540.04 (n=9) &muGSH.(mg protein)-1 (p=0.62), in adults and aged rats
respectively; 76.913.3 (n=5) and 76.642.12 (n=6) nmols thiols.(mg protein)-1 (p=0.94) in adults and aged rats respectively. A.
brasiliensis treatment, however, increased the thiol levels in the aged animals (83.171.56 ng (mg protein)-1; n=5; p=0.04).
Ageing did not change the activity of the antioxidant enzymes: CAT, 12.900.57 (n=5) and 12.890.74 (n=7; p=0.99)
&mumol.min-1.mg-1; SOD, 1.860.12 (n=4) and 1.700.07 (n=8; p=0.28) units.mg-1; GR, 16.090.67 (n=5) and 14.51.1.05
nmol.min-1.mg-1 (n=6; p=0.26). A. brasiliensis treatment, however, increased the activity of CAT and SOD in aged rats:
17.780.86 (n=5; p=0.001) &mumol.min-1.mg-1 and 2.040.07 (n=6; p=0.002) U.mg-1, respectively.
Conclusions:
It can be concluded that treatment with the aqueous extract of Agaricus brasiliensis protects the brain against the oxidative stress
that occurs during ageing. Possibly, this protection minimizes the occurrence of those disorders associated to ageing that involve
the participation of free radicals.
Keywords: Ageing, Medicinal mushroom, Oxidative stress, Rat brain, Reactive oxygen species
Resumo:21-152
DEPRESSIVE-LIKE AND ANXIOGENIC EFFECTS IN ADULT MICE EXPOSED TO ORGANOPHOSPHATE
PESTICIDES DURING NEONATAL LIFE
Lima C. S. 1; Dutra-tavares A. C. 1; Nunes F. 1; Filgueiras, C. C. 1; Manhes, A. C. 1; Meyer, A. 2; Abreuvillaa Y. 1

1
Departamento de Cincias Fisiolgicas/ IBRAG, UERJ
2
Instituto de Estudos em Sade Coletiva, UFRJ
Objectives:
Aim:Epidemiologic studies suggest that organophosphates (OPs) are etiologically linked to mood disorders and cognitive deficits
and that certain subpopulations, particularly children, may be especially vulnerable. Here, we investigated whether early exposure
to chlorpyrifos (CHLOR) and methamidophos (METH) at doses well below the threshold for systemic toxicity and that cause
only 20% of acetylcholinesterase (ACHE) inhibition elicit behavioral disorders at adulthood.
Methods: From postnatal day 3 to 9 (PN3-PN9), Swiss mice received daily s.c. injections of CHLOR (3mg/kg), METH (1mg/kg)
or DMSO vehicle (CONT). These doses were selected because we have previously shown that they cause small and similar
inhibitions of AChE in brain. At adulthood (PN60) mice were submitted to a battery of tests: the forced swimming test (N= 46),
in which the animal is placed in a cylindrical container with water to swim for 10 min and the tail suspension test (N= 55), in
which the animal is hanged by the tail for 6 min. In both tests, the immobility time is quantified and used as a measure of
depressive-like behavior. Anxiety levels were evaluated by the elevated plus maze test (N= 40) which consists of a suspended
maze with two open arms and two arms enclosed by walls. The time that animals remain in open and closed arms is measured.
Learning/memory was evaluated by the step down passive avoidance (N= 53), in which the animals learn that stepping down
from a platform is followed by a foot shock and on subsequent exposures to the task they stay longer on the safe platform.
Locomotor activity was evaluated by the open field test (N= 51), in which animals are placed in a square box and the traveled
distance in cm is quantified. Results were evaluated by analyses of variance (ANOVAs) on all factors: treatment and sex.
Treatment effects and interactions were followed by post-hoc analyses using Fishers Protected Least Significant Difference tests
(FPLSD). Results: Animals exposed to methamidophos presented an increase in immobility time in the forced swimming test
when compared to control animals (METH vs CONT: 36.89.5 vs 8.364.6; P=0.006), indicating a depressive-like behavior. In
the tail suspension test there were no significant differences between groups. In the elevated plus maze, animals exposed to
chlorpyrifos presented a decrease of total time in the open arms (CHLOR vs CONT, 2.922.4 vs 26.9410.3; P=0.012) and in the
percentage of time in the open arms (CHLOR vs CONT, 2.211.67 vs 8.142.39; P=0.013), indicating an anxiogenic effect.
There were no significant differences in locomotor activity (open field) and in learning/memory (passive avoidance).
Conclusions:
Conclusion: Our results demonstrate that exposure to chlorpyrifos and methamidophos during development elicit behavioral
alterations at adulthood. These results corroborate previous studies that suggest that OP exposure elicit mood disorders and
reinforce the importance of studying long-term effects. Finally, since the doses of methamidophos and chlorpyrifos used in the
present study elicit similar levels of ACHE inhibition (20%), distinct behavioral outcomes associated either to methamidophos
or to chlorpyrifos exposure suggest that ACHE inhibition alone does not fully explain our results.
Keywords: anxiety, behavior, chlorpyrifos, depression, methamidophos
Financial Support: CAPES and CNPq, and grants from FAPERJ and MSSM (Award Number
D43TW000640)
Resumo:21-153
EFFECTS OF 8-PHENYLTHEOPHYLLINE ON MOTOR BEHAVIOR IMPAIRMENT INDUCED BY
HALOPERIDOL, IN MICE
Ribeiro, T. S. ; Arajo, R. S. ; Schiaveto-de-souza. A.

Dpt. of Morphophysiology, Universidade Federal de MS, UFMS
Objectives:
Adenosine plays an important hole on basal nuclei functions; however dopamine and adenosine interactions on these nuclei are
not completely understood. The aim of this study was assess the effects of an A1 selective adenosine antagonist, 8phenylteophylline, on motor behavior impairment induced by the D2 dopamine antagonist receptor, haloperidol, in mice.
: Swiss male mice, 25-35g weight, were initially injected by intraperitoneal route with saline or haloperidol and after 30 minutes
injected by intraventricular route with saline or 8-phenylteophylline (5 and 10nM), performing a total of 6 experimental groups:
(n=8/group): i) saline + saline; ii) saline + 8-phenylteophylline (5nM); iii) saline + 8-phenylteophylline (10nM); iv) haloperidol
(2 mg/kg) + saline; v) haloperidol (2mg/kg) + 8-phenylteophylline (5nM); e vi) haloperidol (2mg/kg) + 8-phenylteophylline
(10nM). After drug administration (60 minutes) the animals were submitted to open field test measuring horizontal (quadrants
crossed - HE) and vertical (rearing - VE) exploration. Gait analysis was performed in a runway platform measuring
anteroposterior (APD) and laterolateral deviation (LLD) between fore hind footprint and by stride length (SL) (mm). Comparison
among experimental groups was performed bu one-way ANOVA followed by Duncan post hoc test. The behavior results to each
experimental group 60 minutes after drug administration were: i) saline + saline: HE=78.388.01; VE=29.133.48;
APD=7.621.85; LLD=4.580.75; SL=2.790.19; ii) saline + 8-phenylteophylline (5nM): HE=109.636.94; VE=24.003.26;
APD=4.480.78; LLD=4.000.42; SL=2.920.07; iii) saline + 8-phenylteophylline (10nM): HE=94.7517.73; VE=11.883.16;
APD=7.061.28; LLD=4.760.66; SL=2.690.06; iv) haloperidol (2mg/kg) + saline: HE=18.504.12; VE=4.631.51;
APD=7.680.69; LLD=5.450.44; SL=2.480.09; v) haloperidol (2mg/kg) + 8-phenylteophylline (5nM): HE=25.385.14;
VE=4.381.24; APD=7.511.45; LLD=5.010.64; SL=2.690.13; and vi) haloperidol (2mg/kg) + 8-phenylteophylline (10nM):
HE=13.756.61; VE=1.130.88; APD=6.231.31; LLD=5.731.16; SL=2.820.38. Groups that received haloperidol (iv, v and
vi) crossed less quadrants than other groups in spite of 8-phenylteophylline application (Duncan, p0.05)
Conclusions:
Antagonism of dopamine receptors by haloperidol produced impairment on animal motor behavior, specifically on horizontal and
vertical exploration, that was not reverted by 8-phenylteophylline an A1 adenosine antagonist receptor in both doses used (5 and
10nM). Apparently, animal gait was not affected by dopamine antagonism.
Keywords: BEHAVIOR, HALOPERIDOL, 8-PHENYLTHEOPHYLLINE, IMPAIRMENT, MOTOR
Financial Support: UFMS and CNPq
Resumo:21-154
EFFECT OF CHRONIC INTAKE OF DIETS RICH IN HYDROGENATED VEGETABLE FAT ON THE
PERFORMANCE OF RATS IN THE ELEVATED PLUS MAZE.
Ferreira, A. K. ; Lins, P. G. ; Ladislau, H. F. L. ; Rocha-de-melo, A. P. ; Borba, J. M. C. ; Pereira-da-silva,

M. S.
Depto. de Nutrio- Lab. de Fisiologia da Nutrio - LAFINNT, UFPE
Objectives:
Aim: Observe the effect of chronic intake of a diet high in hydrogenated vegetable fat on the performance of rats in the elevated
plus maze (EPM).
Methods and Results: Female Wistar rats were fed during gestation and lactation with diets containing 7% soybean oil (control,
C) or 14% of hydrogenated vegetable fat (experimental; E). After weaning (21 days) the male pups of the control (C, n = 18) and
experimental (E, n = 11) group were fed the same diets of their respective mothers. The animals were weighed at 1 st, 7th, 14th, 21st
and 30th day of life. At 35 days of age, each animal of both groups was put in the center of the maze with the head orientated to
one of the closed arms for 5 minutes. The session was filmed by a video camera, installed on the roof of the experimental room,
for subsequent analysis of the following behavioral categories: number of entries into the open and closed arms; total time spent
in the open (OA) and closed arms (CA) and exploration of the segments of the open arms. Pups of the E group showed a lower
weight gain at the 1st (E= 6 1; C= 6,82 - 0,852, medianinterquartile) and at the 7th (E= 14 2,75; C= 15,83 4,9925) lactation
day when compared to the group C group. There was no significant difference in the number of entries in the OA and in the CA
between the groups (OA: E= 7,90 0,89 and C= 7,61 0,80; CA: E= 10,36 1,08 and C= 10,5 0,80; Mean SEM). The same
was observed about the time spent (sec) in the arms of the maze (OA: E= 73 12,94 and C= 110 15; CA: E= 127 17,48 and
C= 153 12; Mean SEM ). On the other hand, the frequency of visits into the different segments of the OA was increase for the
segment 2 of the E group when compared to the C group ( E= 4,0-2,75 and C= 2,5-1,0; median-interquartile). No significant
differences were observed for segment 1 which correspondes to the nearest third of the central area and the segment 3 which
corresponds to the extremity of the arm.
Conclusions:
Conclusion: The chronic intake of a high fat diet based on the hydrogenated vegetable fat induced an increase of the weight gain
and seemed to promote a greater impulsivity / lower risk assessment in young rats.
Keywords: Chronic Intake, Elevated Plus Maze, Hydrogenated Vegetable Fat, Rats
Financial Support: PROSPESQ/UFPE
Resumo:21-155
SEROTONIN-INDUCED DIPSOGENIC RESPONSE IN PIGEONS (COLUMBA LIVIA): MEDIATION BY 5-HT 1/2
AND ALFA-1 RECEPTORS
dos Santos, T. S. 1; Melleu, F. F. 1; Souza, V. D. 1; Marino-neto, J. 1,2

1
DEPT. OF PHYSIOLOGICAL SCIENCES, CCB, UFSC
2
INSTITUTE OF BIOMEDICAL ENGINEERING, EEL-CTC, UFSC
Objectives:
Aim: In mammals, interactions between noradrenergic and serotonergic mechanisms have been shown to significantly modulate
several motivational states, including thirst and hunger, as well as the expression of sleep/waking signs. In birds, we have shown
that central injections of noradrenaline or serotonin (5-HT) can differentially affect drinking, feeding and sleep-like behaviors
(SLB). Intracerebroventricular (ICV) 5-HT injections in free-feeding/drinking (FF/D) pigeons (C. livia), besides hypophagy,
evoke intense drinking and SLB, which are negatively correlated to Fos-like activation in serotonergic brainstem neurons (Behav
Brain Res, 220:173, 2011). Central (ICV, intra-raphe) injections of 8-OH-DPAT (a 5-HT1A receptor agonist that reduces 5-HT
neuronal activity by autoreceptor activation) also elicit intense drinking and SLB, suggesting the existence of a tonic, 5-HT1Amediated, inhibitory influence of 5-HT circuits on thirst and sleep in this species.

Methods and Results: This study examined the effects of ICV injections of 5-HT (150 nmol) on drinking, feeding and SLB of
FF/D pigeons, pretreated ICV with metergoline (MET, 5-HT1/2 receptor antagonist; 15, 50,150 nmol), WAY-100635 (WAY, 5HT1A receptor antagonist; 0.1, 0.3, 1 nmol), MM77 (an antagonist equally potent at post-synaptic 5-HT1A receptors and at 1adrenoceptors; 23, 69 nmol) or prazosin (1-adrenoceptors antagonist: 0.9, 2.7 nmol). MET injections completely blocked the
dipsogenic, hypophagic and hypnogenic effects of 5-HT treatments, and produced hyperphagic (but no drinking or sleeping)
responses when given alone, suggesting that all these effects are mediated by 5-HT1/2 receptors. WAY failed to affect the 5-HTinduced drinking, feeding or sleep responses, suggesting that its effect is not 5-HT1A receptor related. MM77 and prazosin dosedependently decreased 5-HT-induced drinking suggesting that dipsogenic responses are depends of that activation of 1
adrenoceptors. Sleep responses after 5-HT were potentiated by prazosin but were contradictorily affect by MM77, suggesting that
1 adrenoceptor can participate of sleep modulation.
Conclusions:
Conclusion: These results showed the participation of 5-HT1/2, probably not 5-HT1A, receptors in the serotonin control of
drinking behavior in pigeons. And besides, as well as in mammals, indicate the important interation between serotonergic and
noradrenergic system in control of drinking behavior in bird, that can indicated relationships that are shared functional traits of
this circuits in amniotes.
Keywords: Drinking behavior, Serotonin, 5-HT1A receptor, Alfa-1 receptor, pigeon
Financial Support: Sources of research support: Capes, Cnpq, Fapesc
Resumo:21-156
AN INVERSE RELATIONSHIP BETWEEN ANNEXIN V AND TNF-ALPHA LEVELS IN CHRONIC STABLE
PATIENTS WITH SCHIZOPHRENIA
Pfaffenseller, B. 1,2,3; Francesconi, L. P. 2,3; Ceresr, K. M. 2,3; Mascarenhas, R. 2,3; Stertz, L. 1,2,3; Gubert, C.
M. 2,3; Lersch, C. 2,3; Gama, C. S. 2,3; Belmonte-de-abreu, P. 2,3
1
PPG Bioqumica/Universidade Federal do Rio Grande do Sul, UFRGS
2
Laboratrio de Psiquiatria Molecular/HCPA, HCPA
3
INCT-Translacional em Medicina, INCT-TM
Objectives:
Apoptotic processes may alter the neuronal network and are associated with the pathogenesis of several neurodegenerative
diseases, such as schizophrenia. The high incidence of inflammatory processes is also known in schizophrenic patients. Annexins
belong to a family of proteins that bind both calcium and phospholipids and form calcium-dependent ion voltage channels within
the lipid bilayer, regulating fagocitosis, cellular signaling, apoptosis, leucocitic migration, proteins like phospholipase A2 and
several components of inflammation, such as cytokines. Among these, tumor necrosis factor (TNF-alpha) is a cytokine
stimulating systemic inflammation and acute phase reactions. The objective of this study was to evaluate annexin V and TNFalpha serum levels in patients diagnosed with schizophrenia and controls.
The study sample consisted of 38 patients diagnosed with schizophrenia from the Hospital de Clnicas de Porto Alegre matched
by age and sex. All patients and controls signed an informed consent form. Diagnosis was given by a trained psychiatrist and
according to the SCID-IV (Structured Clinical Interview for DSM-IV) and Opcrit 4.0. Disease severity was measured by BPRS
(Brief Psychiatric Rating Scale) and illness duration recorded in years. Serum annexin V and TNF-alpha levels were assessed by
enzyme-linked immunosorbent sandwich assay. There was a significant difference in annexin V and TNF-alpha levels between
patients and controls by Wilcoxon T test (p
Conclusions:
The high levels of annexin V in patients diagnosed with schizophrenia can be responsible for the diminished levels of TNF-alpha
due to its anti-inflammatory action. The identification of the role of inflammatory and anti-inflammatory mediators in
schizophrenia can lead to targeted interventions to reduce disease progression, apoptosis and handicap associated to the illness.
Keywords: ANNEXIN V, TNF-ALPHA, SCHIZOPHRENIA, APOPTOSIS, INFLAMMATION
Financial Support: CNPq, FIPE-HCPA
Resumo:21-157
M-TRIFLUOROMETHYL-DIPHENYL DISELENIDE PROTECTS AGAINST GA-INDUCED SEIZURES AND
REACTIVE OXYGEN SPECIES PRODUCTION IN PUP RATS.
Quines, C. B. ; Magni, D. V. ; Bruning, C. A. ; Nogueira, C. W.

Objectives:
Glutaric acidemia type I (GA-I) is an inherited organic acid cerebral disorder caused by the impairment of mitochondrial enzyme
activity glutaryl-CoA dehydrogenase, which gives rise to an accumulation of glutaric acid (GA) in body fluids and brain tissue of
affected patients. Clinical manifestations of GA-I are predominantly neurological, including generalized convulsions. mTrifluoromethyl-diphenyl diselenide (CF3) is an organoselenium compound that has showed interesting pharmacological
properties, such as antioxidant and anticonvulsant activities. In this way, we evaluated whether administration of CF3 protects
against GA-induced seizures and reactive oxygen species (ROS) production.
Pup male Wistar rats were anesthetized and placed in stereotaxic apparatus for cannula insertion into the striatum (coordinates
relative to bregma: AP, 0 mm; ML, 3.0 mm; V, 3.0 mm from the dura). The experiments were performed 3 days after surgery
when animals did not show any sign of pain, infection or discomfort. Pup rats (21 days-old) were divided into 4 groups of 12
animals each. A control group administered with oil + saline, another group treated with CF3 + saline, a third group administered
with only oil + GA, and a fourth group administered with CF3 + GA. CF3 compound (50 mg/kg) or oil (vehicle) were infused by
intragastric gavage 30 min before the intrastriatal administration of GA (1 L, 1.3 mol/striatum) or saline. Immediately after the
intrastriatal injections we observed the latency and duration of seizures for 20 min. In the present investigation, we verify that
administration of CF3 increased significantly the latency for the first convulsive episode induced by intrastriatal injection of GA.
Besides, the treatment with CF3 decreased significantly the duration of seizures. In addition, the analysis of ROS demonstrated
significantly greater levels in the group administered only with GA when compared to the group administered with CF3 + GA.
Conclusions:
Previous administration of CF3 in pup rats from 21 days-old gave a protective effect against GA-induced seizures, demonstrated
by an increase of latency and a reduction of duration of convulsive episodes. Besides, the pre-administration of CF3 decreased
the GA-induced ROS levels. Although our results are promising, it is important to understand the mechanisms involved in the
protective effect of CF3 on GA-induced seizures for indicating a new anticonvulsant therapy for treatment of this organic
acidemia.
Keywords: Glutaric acid, m-Trifluoromethyl-diphenyl diselenide, seizures, reactive oxygen species, pup rats
Financial Support: CAPES/CNPq
Resumo:22-031
EFFECTS OF DIFFERENTS PROTOCOLS OF PHISICAL TRAINING ON FUNCTIONAL AND
MORPHOLOGYCAL ASPECTS OF RATS SUBMITED TO MEDIAN NERVE CRUSH INJURY
Neves, J. D. 1; Camelier, F. S. 1; Silva, S. A. D. 1; Arsego, N. R. 1; Marcuzzo, S. 1; Xavier, L. L. 2; Faccioniheuser, M. C. 1

1
Departamento de Cincias Morfolgicas, UFRGS
2
Laboratrio de Morfologia, PUCRS
Objectives:
The aim of the present study was to compare the effects of balance and coordination, repetition and the combination of these two
types of training protocols on functional recovery and morphometric outcome of the median nerve after crush injury.
46 male Wistar rats (3 months) were used. Procedures were approved by the Ethical Committee at the UFRGS. Animals were
randomly divided in five groups: sham sedentary (Sh-s, n = 8) - operated rats, without median nerve crush and unexercised;
lesioned sedentary (LSE, n = 9) - rats with median nerve crush and unexercised; repetition (LR, n = 9) - rats with median nerve
crush and repetition training; balance and coordination (LBC, n = 10) - rats with median nerve crush and balance and
coordination training; balance and coordination + repetition (LBCR, n = 10) - rats with median nerve crush and balance and
coordination + repetition training. For the surgical procedures, animals were anesthetized and nerve crush injury was performed
with 1 mm hemostatic forceps for 30s. Four days after the surgery, the animals from the LR, LBC and LBCR groups began
specific training for 6 weeks. During training, the animals underwent the behavioral assessment with the footfault test (FT). Fixed
samples of the distal portion of median nerve were prepared for electron microscopy and cut at 1 m for assessing axonal density,
average myelinated fiber area, myelin sheath thickness, myelinated fiber diameter and axonal diameter. The results were analyzed
by one-way ANOVA for functional test and repeated measures ANOVA for morphometric analysis followed by post hoc
Duncans with software Statystical. Footfault test revealed that the number of right forelimb errors was significantly decreased in
the Sh-s group 3 days postlesion, compared with all the lesioned groups: LSE (p
Conclusions:
The data showed evidence that balance and coordination training accelerate nerve regeneration after experimental traumatic
injury, spite footfault test not show differences between the treatments.
Keywords: crush injury, peripheral nerve regeneration, training protocol
Financial Support: CAPES and Cnpq
Resumo:22-032
A POSSIBLE ROLE FOR MMP-9 IN THE RAT RETINOTECTAL SYSTEM.
Oliveira-poleto, A. L. ; Cabral-miranda, F. ; Campello-costa, P. ; Serfaty, C. A. ; Oliveira-silva, P.

Department of Neurobiology, Institute of Biology, UFF
Objectives:
During postnatal development retinal projections must reach correctly their targets in the superior colliculus (SC) in order to
establish functional synapses. In this context, inappropriate axons are retracted and eliminated during the critical period of
synapses formation. These events guarantee the correct representation of the visual field in central visual nuclei. Matrix
metalloproteinases (MMPs) are a family of enzymes that degrade most of the extracellular matrix components, creating a
permissive environment for axonal growth and synaptic plasticity. MMPs activity is regulated by the tissue inhibitors of
metalloproteinases (TIMPs) and imbalance of MMPs/TIMPs during development impairs the formation of topography of neural
circuits. Previous data of our lab have shown that MMP-9 activity decreases during SC development and the inhibition of
gelatinases in newborn animals harms the refinement of neural circuitry in this structure. Our objective was to search for the
impact of blockade of MMP-9 activity, using SB-3CT (Calbiochem) in a concentrationof 648M, which is recognized to be
specific to inhibit this molecule without interfere with others MMPs activity.
Newborn Lister Hooded rats were implanted with ELVAX containing SB-3CT and Horseradish Peroxidase was injected in
temporal retina of animals at PND6. Animals were submitted to a perfusion trough heart at PND7 with saline 0.9% followed by
aldehydes, afterwards the brains removed and left in sucrose solution for 24h. 40 coronal sections were obtained in a cryostat
and stored in 0.1M phosphate buffer. Then sections were transferred to 0.1M acetate buffer and incubated with TMB and H2O2.
Analysis was made using a Zeiss microscope. Microscope analysis revealed that retinotectal projections in SC of animals treated
with SB-3CT appeared expanded and diffused in comparison with control rats, disrupting the cluster formation pattern observed
during development. This result led us to believe that the specific inhibition of MMP-9 activity in the first postnatal week might
delay or even abrogate the refinement of topography in rats visual system.
Conclusions:
Our result suggests that MMP-9 activity have a key role in the refinement of retinotectal projections in SC, however further
analyses must be made to better elucidate the contributions of MMP-9 to neural plasticity.
Keywords: mmp-9, retinotectal, plasticity, development
Financial Support: Proppi/UFF & Faperj
Resumo:22-033
INTERFERON GAMMA INFLUENCES SYNAPSE ELIMINATION PROCESS WITHOUT AFFECTING THE
PERIPHERAL REGENERATION AFTER INJURY
Victrio, S. C. S. ; Cartarozzi, L. P. ; Oliveira, A. L. R.

Depto. Anatomia/ UNICAMP, UNICAMP
Objectives:
Recent studies have indicated that the major histocompatibility complex (MHC I) expression by injured neurons and surrounding
glia plays an important role for neuronal repair, influencing the synaptic plasticity process. Additionally, interferon gamma
expression (IFN), a potent cytokine involved in MHC I expression, is upregulated after trauma. Taking into account that little is
known about the functional importance of IFN in the central nervous system (CNS) and peripheral nervous system (PNS) after
peripheral injury, the aim of the present work was to investigate the process of synaptic elimination in the spinal cord as well as
the motor recovery in IFN-/- mice, following sciatic nerve axotomy.
One week after transection and two weeks after crush lesion, the animals were sacrificed and had their lumbar spinal cord and
sciatic nerves processed for immunohistochemistry, Western blotting (WB) and electron microscopy. Newborn mice astrocyte
primary cultures were established in order to study the astrocytic respose in the absence of IFN expression. The lack of IFN
decreased the expression of MHC I (IFN-/-, 2.720.53; C57BL/6J, 5.210.48;ratio ipsi/contralateral; integrated density of pixels
x103; p0.05). The synaptophysin immunolabeling increased in the microenvironment surrounding large motoneurons of mutant
mice (IFN-/-, 4.410.6; C57BL/6J, 2.540.23; ratio ipsi/contralateral, integrated density of pixels; p0.05) and GFAP (IFN-/-,
2.270.24; C57BL/6J, 2.330.13; ratio ipsi/contralateral, integrated density of pixels; p>0.05) reactivity were equally increased
in both strains. In vitro, the astrocytes demonstrated similar GFAP levels, but the proliferation rate was higher in the wild type
mice. In the crushed nerves (distal stump), neurofilaments and p75NGFR immunolabeling were upregulated in mutant mice as
compared to the wild type, although it did not result in any improvement of locomotor recovery.
Conclusions:
The present results show that the lack of IFN affects the synaptic elimination process in the spinal cord. Such changes, however,
do not delay the peripheral nerve regeneration after nerve injury.
Keywords: interferon gamma, MHC class I, motoneuron, spinal cord
Resumo:22-034
STUDY OF SYNAPSES IN THE TEMPORAL CORTEX DURING AGING
Merlo, S. ; Nakayama, A. B. ; Brusco, J. ; Rossi, M. ; Carlotti Jr, C. G. ; Moreira, J. E.

FACULDADE DE MEDICINA DE RIBEIRO PRETO, FMRP USP
Objectives:
Analyze the temporal cortex of humans and rats: the number of excitatory, inhibitory, and electric synapses, the site of
postsynaptic contacts, the number of synaptic vesicles per terminal, and the expression of the proteins alpha-synuclein and
ubiquitin following the size and features of the lipofuscin granules.

Samples of temporal cortex of human subjects with different ages (20-28, 37-41 and 50-55 years, n=4/5) were collected from
patients with epilepsy who underwent temporal lobectomy. Samples from Wistar rats of 2, 6, 10 and 12 months were also
collected. Light, confocal, and electron microscopy, and western blots techniques were used as procedures. The data obtained on
lipofuscin granules were coincident with other studies that observed a higher area occupied by this structure in older mammals.
The granules seem to grow in volume, but not in number, with considerable increase of the electron lucid fraction (lipidic). There
was no difference in the alpha-synuclein and ubiquitin expressions between the experimental groups. The synaptic densities were
similar between the groups, as well as the postsynaptic contacts. Increase of the electron dense vesicles in inhibitory synapses,
appeared to be associated with the demand of catecholamines.
Conclusions:
Thal and Schlote (1994) also observed increase of the electron lucid portion and reduction of the electron dense portion with
aging. High levels of oxidative stress are related an increase of reactive oxygen species with aging (Yankner et al., 2008).
Already studies in mammals observed decrease in the number of synapses in older individuals. Huttenlocher (1979) examined the
synaptic density in layer 3 of the middle frontal gyrus, from infants to individuals 90 years of age. The author observed that the
synaptic density was constant in this region between the ages of 16 to 72 years, but declined markedly in individuals 74 to 90
years old. Therefore the results of the present work do not express totally the aging process, because the range of age used in
humans, and rats belong to a young age.
Keywords: AGING, TEMPORAL CORTEX, SYNAPSES, LIPOFUSCIN GRANULES, ELECTRON MICROSCOPY
Financial Support: FAEPA, CNPq, FAPESP
Resumo:22-035
DENDRITIC SPINE DENSITY IN HIPPOCAMPUS FOLLOWING NEONATAL HYPOXIA-ISCHEMIA IN RATS
SUBMITTED TO DAILY ENVIRONMENTAL ENRICHMENT
Deniz, B. F. 1; Rojas, J. J. 1; Miguel, P. M. 1; Diaz, R. 1; Hermel, E. E. S. 1; Achaval, M. E. 1; Netto, C. A. 1;

Pereira, L. O. 1
1
Departamento de Cincias Morfolgicas, UFRGS
2
Departamento de Bioqumicas, UFRGS
Objectives:
Previous studies from our group showed the neuroprotective effects of environmental enrichment (EE) in rats submitted to the
neonatal hypoxia-ischemia (HI); however it didnt cause any change in lesion volume in the hippocampus. The aim of this study
was to investigate possible changes in dendritic spines density in the dorsal CA1 hippocampus region of rats submitted to HI and
exposed to EE.
Seven-day old Wistar male rats were submitted to HI procedure which consisted in permanent occlusion of the right common
carotid artery and posterior exposure to 90 minutes to hypoxic atmosphere. The pups were divided into four experimental groups,
with 5 animals per group: control maintained in standard environmental (CTSE); control exposed to environmental enrichment
(CTEE); submitted to hypoxia-ischemia and maintained in standard environmental (HISE); submitted to hypoxia-ischemia and
exposed to environmental enrichment (HIEE). At the 22 days old the EE started and continue during 9 weeks, being performed 6
days per week, 1 hour per day, during 9 weeks. Two days after the end of EE the animals were transcardially perfused with
fixative solution and their brains were submitted to Golgi method procedure. The morphological analysis was performed using a
camera lucida coupled to an optic microscope that allows simultaneous visualization and design of the sample. For each CA1
pyramidal cell the morphologic analysis included the dendritic spines number in 20 m of the selected most lateral tertiary
dendrites on the apical tree. From each rat, ten different dendrites were analyzed, five per hemisphere, one per neuron. For
statistical comparisons, the means of each rat were used and, after, a two-way analysis of variance (ANOVA) was conducted.
There was effect referent to the lesion (F (1,16) = 39.340, p < 0.01; F (1,16) = 55.104, p < 0.01) and environment (F (1,16) =
13.256, p < 0.01; F (1,16) = 20.680, p < 0.01) both in the left and right hemisphere, respectively. Hypoxic-ischemic maintained in
SE animals had lower dendritic spines densities when compared to both control groups (p
Conclusions:
These data of dendritic spines indicated a completely recovery in the left hemisphere and a partially recovery in the right
hemisphere after environmental stimulation in animals submitted to hypoxia-ischemia. Maybe these results could explain a
possible morphological effect to explain the functional recovery, previously published, consequent to environmental enrichment
in hypoxic-ischemic animals
Keywords: environmental enriched, hypoxia-ischemia, dendritic spine density
Financial Support: CAPES, CNPQ and FAPERGS.
Resumo:22-036
EFFECTS OF AXOTOMY ON SATELLITE GLIAL CELL ULTRASTRUCTURE IN FROG DORSAL ROOT
GANGLIA.
Rigon, F. ; Faccioni-heuser, M. C. ; Partata, W. A.

instituo de cincias biolgicas da sade, UFRGS
Objectives:
Peripheral nerve lesion lead to morphological changes in the cell bodies of the neurons of origin. Apoptosis and necrosis
contribute to neuronal death in this experimental condition. Signs of satellite glial cell (SGC) death also occur after peripheral
axotomy and the Tunel-positive cells are already present at day four, peaking at two months. Since the changes of SGC following
peripheral axotomy are not clearly understood, the present study was carried out to determine the effect of sciatic nerve
transection (SNT) on the ultrastructure of SGCs located in the dorsal root ganglia (DRG). Since the amphibian brain shares nearly
all the typical properties of the amniote brains, frogs were used as an experimental model in this study to offer a phylogenetic
basis of the effects of peripheral axonal injury.
Specimens of adult male frogs Rana catesbeiana weighing 100-200g, obtained from RANASUL (Imb, RS), were divided in two
experimental groups (six animal/group): control (animals which did not undergo surgical manipulation) and SNT (the sciatic
nerve was exposed and transected approximately 5 mm distal to the sciatic notch). These animals were sacrificed 3 days after the
surgical procedure, since signs of mammal SGC death in this period were previously reported. After the sacrifice the DRGs of the
sciatic nerve ipsilateral to the lesion were dissected out and fixed in a 4% paraformaldehyde and 2.5% glutaraldehyde solution in
0.1M phosphate buffer (PB), pH 7.4, at room temperature. The ganglia were postfixed in osmium tetroxide in PB for one hour.
Afterwards, the sections were dehydrated in series of alcohol and acetone and embedded in araldite (Durcupan ACM, Fluka).
Semithin (900 nm) and ultrathin (70 nm) sections were obtained using a diamond knife (Drukker, Netherlands). The ultrathin
sections were mounted and stained with 2% uranyl acetate followed by lead citrate and examined by transmission electron
microscopy (JEM 1200 EX II). While in ganglionic neurons the earliest response to axotomy is the presence of irregularities in
the nucleus and perinuclear cytoplasm, no discontinuities were found in the cytoplasmic membrane and nucleolemma of the SGC
3 days after SNT. No changes in electron density and chromatin condensation were observed. There were more irregularities in
the SGC prolongation. These processes appeared more plicated than in the normal cell. Many free ribosomes, rough endoplasmic
reticulum and mitochondria were observed. No swelling was found in these organelles. They were scattered within the cytoplasm
and the perinuclear region showed increase in the amount of filaments. While a chromatolytic phenomenon was found in some
neurons, the SGCs did not show this characteristic in this time frame.
Conclusions:
These results showed that sciatic nerve transection did not induce SGC death at 3 days. Although frogs have a lower metabolic
rate in the nervous system than mammals, the fine structure morphology of the SGC was similar to that described for mammals.
Since it was suggested that SGCs support the mammalian neurons metabolically, the results obtained up to now clearly indicate
that SGCs are capable of actively synthesizing the proteins necessary to support the dorsal root neurons in the regenerative
process in the frog nervous system.
Keywords: axotomy, dorsal root ganglia, satellite glial cell
Resumo:22-037
ABSENCE OF TOLL LIKE RECEPTOR 2 EXPRESSION INFLUENCES THE INITIAL STEPS OF PERIPHERAL
NERVE REGENERATION AFTER AXOTOMY
Leite, G. A. ; Freria, C. M. ; Oliveira, A. L. R.

Department of Anatomy, Cell Biology and Physiology and Bioph, UNICAMP
Objectives:
Peripheral nerve regeneration after axotomy is a complex process that involves different signaling pathways. Following crush or
transection of a nerve, the Wallerian Degeneration (WD) process develops distally to the lesion site, mostly at the distal stump.
Such process is characterized by the fragmentation of myelin sheath, macrophage recruitment and Schwann cell proliferation.
Concurrently to the distal changes, retrograde phenomena takes place in the spinal cord, at the motoneuron cell body and
surrounding microenvironment, activating microglial cells and astrocytes in the central nervous system (CNS). Recent studies
have shown that glial cells, including Schwann cells, astrocytes and microglia express different toll like receptors (TLRs). The
TLRs are transmembrane recognition receptors, which identify pathogens and initiate a local immune response. Such receptors
might influence several events after injury and repair process within the nervous system. In this context, the present work aimed
at investigating the influence of knocking out the TLR2 on the peripheral nerve regeneration process after sciatic nerve axotomy.
C57BL/6J (n=5) and C57BL/6J TLR2-/- (n=5) mice were subjected to unilateral sciatic nerve crush. Two weeks after surgery the
mice were sacrificed and perfused transcardially with fixative (10% formaldehyde in 0.1M PBS). Contralateral and lesioned
sciatic nerves were obtained, frozen and cryostate sectioned. Tissue samples from both groups were subjected to
immunohistochemistry with anti-neurofilaments, anti-laminin I, anti-collagen IV and anti-p75 NGFR antisera. Gait recovery after
sciatic nerve crush was performed up to five weeks post lesion in another set of control and knockout mice (n=5 for each group),
by using the CatWalk system (Noldus Information Technology). The results showed greater expression of p75NGFR,
neurofilaments and collagen IV in the control group as compared to knock out mice. However, regardless such protein expression
changes in the acute phase post lesion, no significant differences were observed in the functional analysis (four weeks postlesion: C57BL/6J 85,19% 12,30%, C57BL/6J TLR2-/- 64,52% 7,66%; five weeks post-lesion: C57BL/6J 72,25% 10,72%,
C57BL/6J TLR2-/- 71,00% 9,21%; average percentage of motor recovery standard error of mean).
Conclusions:
Altogether, the present results indicate that the absence of TLR2 negatively influences the initial response to injury, possibly by
postponing the nerve microenvironment reorganization. Nevertheless, as seen by the functional recovery, such initial delay does
not affect the axonal growth itself as well as the muscle reinnervation.
Keywords: Toll like receptor 2, Peripheral Nerve Regeneration, Wallerian Degeneration, Axotomy, Functional recovery
Financial Support: FAPESP - Fundao de Amparo Pesquisa do Estado de So Paulo.
Resumo:22-038
REELIN EXPRESSION DURING DEVELOPMENT OF RETINOCOLLICULAR PATHWAYS: MODULATION
THROUGH TRYPTOPHAN NUTRITIONAL RESTRICTION
Antonioli-santos, R. 1; Gonzalez, E. M. C. 1; Hedin-pereira, C. 2; Oliveira-silva, P. 1; Campello-costa, P. 1;

Serfaty, C. A. 1
1
Depto. de Neurobiologia/ Instituto de Biologia, UFF
2
Depto. de Anatomia/ Instituto de Cincias Biomdicas, UFRJ
Objectives:
During postnatal development, neural circuits are estabilished according to their interaction with the environment and also the
availability of nutrients. We studied the role of reelin an extracellular matrix glycoprotein able to affect several steps in brain
development, from neuronal migration and positioning to dendritic maturation and synaptic plasticity on the development of
retinocollicular connections in both normal Lister Hooded rats and rats with decreased levels of brain serotonin through a
nutritional restriction of tryptophan (Try-), an essential aminoacid acquired exclusively by food intake and the metabolic
precursor of serotonin.
As shown in our previous results, the distribution of uncrossed retinocollicular terminal fields was examined following
intraocular injections of horseradish peroxidase (HRP 30%) in aldehyde-fixed, free-floating sections (40m) through the superior
colliculus at the tenth postnatal day (PND10). The neonatal tryptophan restriction alters the retinocollicular topographical
refinement, suggesting a delay in axonal elimination, associated to a decrease of neural plasticity (Exp. Neurol. 211:441, 2008).
In the present study, we performed western blot analysis to evaluate the content of reelin in the visual layers of the superior
colliculus. Reelin was differentially expressed in the collicular visual layers during postnatal development: low levels were found
at PND5 followed by a transient increase over the second postnatal week, reaching the highest expression at PND14. After the
third postnatal week, reelin expression showed a slight decrease and remained stable even after the end of the critical period
(PND28), when rats reach an adult like profile of retinotectal connections. Moreover, the immunofluorescence analysis showed
reelin-positive cells at PND10 with immunoreactivity predominantly located at perineuronal sites. Animals fed with a tryptophan
restricted diet (Try-) from the day of birth until PND10, showed an increase in reelin expression, as compared to control groups
fed with a regular rodent diet.
Conclusions:
The present results indicate the involvement of reelin in the specificity of retinocollicular connections during initial stages of
postnatal development. Also, reelin must be involved on the disruption of retinocollicular topography induced by the low levels
of tryptophan, possibly enabling the stabilization of inappropriate transitory synaptic contacts.
Keywords: reelin, retinocollicular pathway, tryptophan restriction, serotonin
Financial Support: CAPES, FAPERJ, CNPQ, PRONEX, PROPPI/UFF
Resumo:22-039
RELATIONS BETWEEN AGING AND NERVE CELLS IN RATS BRAIN - STEREOLOGICAL STUDY.
Correia, M. H. 1; Cruz, R. S. 1; Janeiro, V. 2; Melo, S. R. 1

Depto. de Cincias Morfolgicas - Univ. Est. de Maring-Pr, DCM - UEM
2
Depto. de Estatstica - Univ. Est. de Maring-Pr, DES - UEM
Objectives:
The neurons and glial cells represent the cells of nervous tissue that together provide the functions of the nervous system, the
relations with neurodegenerative diseases, aging and quantity of these cells has been estimated, however there is no consensus in
the literature. For these reasons, the main purpose of this study was to estimate the density (Vv) of neurons and glial cells in
normal rats brain at different ages.
This experiment used 20 male Wistar rats (Rattus norvegicus) at 10, 14, 18 and 24 months of age, five rats in each group. The
animal was perfused with Zamboni solution, and then the brain was removed, fixed in the same solution, processed in alcohol and
xylene series and embedded in paraffin, which was cut with six micrometers. For each animal three sections were processed with
the technique Picro Sirius Red, and fifteen fields were analyzed by stereological technique. We used video microscope coupled to
a video camera and monitor. Quantification of cells was achieved with the determination of neurons Vv (Vv [neu]) macroglia Vv
(Vv [mac]) and microglia Vv (Vv [mic]). Neurons were distinguished from glial cells based on shape and size of the nucleus and
were considered for the quantification the points from test system (M42) which touched in the nucleus. It was observed that Vv
[neu] in the 10m group was 4.76 in the 14m group was 3.81, in 18m group was 2.38 and was 2.86 in the 24m group. In the Vv
[mac] observed that the 10m group was 6.83, in the 14m group was 6.51, in the 18m group was 7.94 and was 6.98 in the 24m
group. And finally the Vv [mic] in the 10m group was 2.06, in the 14m group was 1.90, in 18m group was 2.38 and was 2.70 in
24m group. The analysis allowed us to infer that the Vv [neu], Vv [mac] and Vv [mic] in the analyzed groups was not statistically
significant, p> 0.05.
Conclusions:
This result shows that aging is not changing the number of neurons and glia. These results lead us to question what factors are
related to this stability in rats and whether this could be applied in human brain.
Keywords: Aging, Macroglia, Microglia, Neurons, Stereology
Financial Support: Fundao Araucria
Resumo:22-040
PNEUMOCOCCAL MENINGITIS: OLFACTORY ENSHEATHING CELL AS PUTATIVE SAFE HOST CELLS FOR
STREPTOCOCCUS PNEUMONIAE
Macedo-ramos, H. 1; Carvalho, L. A. 1; Teixeira, L. M. 2; Cavacante, L. A. 1; Baetas-da-cruz, W. 1,3

1
Laboratrio de Neurobiologia do Desenvolvimento/IBCCF/UFRJ, IBCCF/UFRJ
2
Instituto de Microbiologia/UFRJ, IMPG/UFRJ
3
Campus Maca/UFRJ, Campus Maca/UFRJ
Objectives:
Pneumococcal Meningitis (PM) is an infectious disease caused by the gram-positive diplococcus Streptococcus pneumoniae (Sp).
PM can affect people of any age but is more commonly diagnosed in children in the first two years of life. It is usually believed
that PM induced by the intra-nasal route involves bacteremia before bacteria entry into the brain. The best demonstration of an
alternative route was that in which a strain of Sp that fails to survive in the blood stream was instilled into the nasal cavities and
recovered in the olfactory nerve and bulb as well as in the whole brain (Proc. Natl. Acad. Sci. U.S.A. 100:14363, 2003).
Olfactory ensheathing cells (OECs) comprise a special glial type derived from progenitors in the olfactory mucosa that ensheath
olfactory receptor axons. Axonal bundles extend toward the cribiform plate and enter the brain via the olfactory phila, thus,
providing a potential route for access of pathogens to the brain. In a recent paper, we have shown that OECs express MR and are
able to internalize Sp bacteria of the ATCC 49619 strain, by means of a mannan-blocked lectin, possibly MR itself (Neurosci.
Res. 69:308 2011). Based on the above, we decided to investigate possible variations in the expression levels of inducible nitric
oxide synthase (iNOS) in OECs cultures infected with Sp.
OEC cultures were infected by a suspension of living Sp (ATCC49619, American Type Culture Collection 49619) in a ratio of
100:1 bacteria:cells during 3 h, since formation of colonies after exposure, washing and recovery of bacteria from lysed host cells
was not signicantly different after 1, 3 or 5h. To examine the levels of iNOS protein expression, we used a monoclonal antibody
against the iNOS (anti-iNOS) and bacteria were visualized with a polyclonal antibody (anti-pneumococcus) or Sytox green, an
optimal stain for bacterial DNA. Our results showed a significant decrease in the levels of iNOS expressed in OECs infected by
Sp when compared with (uninfected) controls.
Conclusions:
These results support our hypothesis of a prospectively important role of OECs as a host cell during the bacterial invasion of the
brain via the olfactory route.
Keywords: innate immunity, pattern recognition receptors (PRRs), inducible Nitric Oxide Synthase (iNOS), pathogen-associated
molecular patterns , pneumococcal meningitis
Financial Support: FAPERJ/CNPq
Resumo:22-041
REGION-SPECIFIC NEURONAL BINDING OF A OLIGOMERS INJECTED CHRONICALLY INTO THE

CEREBRAL VENTRICLE OF ADULT MONKEYS.
Forny-germano L. 1; Lyra N. M. S. 1; Graa, D. F. 1; Brito-moreira J. 1; Bomfim, T. R. J. D. S. 1; Munoz, D.

P. 3; Ferreira S. T. 1; Houzel J. C. 1; de Felice. F. G. 1
1
LDN, Instituto de Bioquimica Mdica, UFRJ, UFRJ
2
LFN, Instituto de Cincias Biomdicas, UFRJ, UFRJ
3
Queen's Centre for Neuroscience Studies, Queen's University, Queen's University
Objectives:
The histopatologial features found in post mortem Alzheimers disease (AD) brains are plaques - extracellular deposits of A
peptide- and neurofibrilary tangles - intraneuronal accumulations of hyperphosphorilated tau protein. However, neuropathological
studies failed to correlate plaque levels with clinical symptoms. The current hypothesis for AD is that soluble oligomers of the A
peptide (known as Amyloid -Derived Diffusible Ligands or ADDLs) accumulate in the brain and act as potent neurotoxins,
leading to the functional breakdown of specific synaptic circuits. Recent in vitro studies showed that, after binding to synaptic
sites, ADDLs can induce tau hyperphosphorylation and oxidative stress (Neurobiol. Aging 29:1334, 2008). Transgenic mice
expressing high levels of tau and APP (APPsw-tauvlw) shown that accumulation of oligomeric A, but not of total amyloid
plaque, is an early event that faithfully predicts the selective pattern of regional neuronal vulnerability and closely correlates with
the extent of neuronal cell loss (J Neuropathol Exp Neurol 70; 360, 2011). Previously work from our group used chronic
injections of ADDLs into the lateral ventricule of rats to investigate the regionalization of ADDLs accumulation in rat brain Here,
we extend this model to a non-human primate, investigating whether biochemically characterized A oligomers injected
chronically into the lateral ventricle of adult monkey may alter the levels of hyperphosphorilated tau protein, and bind selectivity
to neurons from specific brain regions.
An intraventricular canula was implanted into the lateral ventricle of three mid-aged (~9 years of age) cynomolgus monkeys
(Macaca fascicularis, weight 4.7-7.0Kg). Every 3 days for 21 days, monkeys received 100g of freshly prepared ADDLs. One
animal was used as control. For immunohistochemistry, the fixed brains (3 ADDLs, 1 sham) were frozen-cut into 40m sections
and processed for the detection of ADDLs, phospho-tau Ser396 or DAPI (nuclei). The ratio of ADDLs+/DAPI+ cells was
counted and compared across selected brain regions. Cellular levels of ptau were compared using NIH Image Java to measure
pixels per area. Our results showed that ADDLs could diffuse from the ventricle to the cerebral parenchyma and bind to cells
with typical neuronal morphology thoughout the brain. However, ADDLs binding was more frequent in regions involved in
higher cognitive functions. The mean fraction of ADDLs+ cells differed across regions: hippocampus: 3,4%; somatosensory
cortex: 2,5%; frontal cortex: 2,4%, thalamus: 2,3%, amigdalar complex: 1,7%; and striatum: 1,4%. No ADDLs labeling was
observed in the cerebellum. Preliminary results revealed a regionalized increase in ptau levels in ADDLs animals. Amygdala: 7
fold, frontal cortex: 2.5 fold and hippocampus: 1.5 fold. More regions are being quantified and additional analyses of the relation
between ptau levels and A oligomers will be developed.
Conclusions:
Our results suggest that chronic intracerebroventricular injection of A oligomers in adult monkeys is an useful model to study
the fate of such toxins in vivo. ADDLs might selectively target neurons in cognitive-related circuits and elevate ptau levels. Such
a regional specificity of the ADDLs attack may help to explain the specific cognitive impairments characteristic of the early
stages of AD.
Keywords: Ab oligomers, amyloid plaque, Alzheimer Disease, intracerebroventricular injection, tau hyperphosphorilation
Resumo:22-042
DENDRITIC ATROPHY IN HIPPOCAMPAL CA3 NEURONS IN PARALLEL TO CORTICOID AND
ANGIOTENSIN RECEPTORS REDUCTION IN ADULT RATS SUBMITTED TO IN UTERO PROTEIN
RESTRICTION
Lopes, A. D. S. 1; Torres, D. B. 1; A. J. Rodrigues3; J. J. Cerqueira3; J. M. Pego3; A. F. Godinho1; J. A. R.

Gontijo2; N. Sousa3; P. A. Boer1
1
Department of Morphology Biosciences Institute, UNESP
2
Department of Internal Medicine, UNICAMP
3
Life and Health Sciences Research Institute, ICVS
Objectives:
Aims: Corticoids have the ability to modulate hippocampal formation during the fetal period through their action in the
mineralocorticoid (MR) and glucocorticoid (GR) receptors. In this work, our objective was to investigate the effects of
intrauterine protein restriction in the adult hippocampal formation and expression/localization of GR and MR. Additionally, since
angiotensin receptor AT1 has been demonstrated to enhance learning and memory, we also studied its hipocampal expression
Methods: Pregnant rats were divided into two groups: a group was fed with normal diet (NP, 17% of protein n=9), and the other
received low protein diet (LP, 6% of protein n=9). Male body weight was measured in the day of birth. At 16 weeks of age, 4
animals from each experimental group were perfused with saline and brains processed for 3D neuronal reconstruction in GolgiCox stained slices. For each selected neuron, all branches of the dendritic tree were reconstructed at 600X magnification using a
motorized microscope attached to Neurolucida software. Fifteen neurons of regions CA1, CA3 and dentate gyrus (DG) were
studied for each animal. Total length of trees and number of dendritic branches were compared across groups using 1-way
analysis of variance. Other brains were processed for AT1, MR and GR immunohistochemistry (IH) (n=4) and for western
blotting (WB) (n=4).Results: LP offspring presented a significant reduction in birth weight (LP 5.34 0.06 vs NP 6.23 0.05,
n=95, p=0.0001). In adults, 3D analysis showed significant reductions in total length of CA3 basal, but not apical, dendrites
(length: LP 920,51 307,54m vs NP 1264,72 445,95m, p=0,01). On the contrary, CA1 and DG dendritic arborizations were
unaffected. By WB we verified that hipocampal expression of AT1 was 90% reduced in the LP group. IH revealed that GR
expression was decreased in CA1 and CA3 hippocampal regions; no differences were found in MR expression.
Conclusions:
Conclusions: Gestational protein restriction promoted significant length reduction of basal dendrites in hippocampal CA3 neurons
in parallel with reduced GR expression in CA1 and CA3 areas and a massive reduction of AT1. Since the hippocampal formation
is extremely vulnerable to both undernutrition and excessive corticosteroid concentration we suppose that elevated fetal exposure
to maternal corticoids may be related with the genesis of these alterations.
Keywords: FETAL PROGRAMMING, MEMORY, HIPOCAMPPUS, DENDRITIC STEREOLOGY, BEHAVIOUR
Financial Support: FAPESP and CAPES
Resumo:22-043
NUTRITIONAL MODULATION OF OMEGA-3 FATTY ACIDS ALTERS THE TOPOGRAPHICAL FINE TUNING
OF RETINOTECTAL PATHWAYS
Velasco, P. C. 1; Menezes, L. C. 1; Faria-melibeu, A. D. C. 1; Borba, J. M. C. 2; Costa, B. L. S. A. 2; Guedes,

R. C. A. 2; Campello-costa, P. 1; Serfaty, C. A. 1
1
Neurobiologia/Universidade Federal Fluminense, UFF
2
Nutrio/Universidade Federal de Pernambuco, UFPE
3
Fisiologia/Universidade Federal de Pernambuco, UFPE
Objectives:
The development and maturation of sensory systems depends on the correct pattern of connections which occurs during a critical
period when axonal elimination and synaptic plasticity are involved in the formation of topographical maps. Among the
mechanisms involved in synaptic stabilization, essential fatty acids (EFAs), which must be obtained through diet, appear as
precursors of signaling molecules involved in modulation of gene expression and also regulation of neurotransmitter production
and release. Omega-3 fatty acids, such as DHA are considered EFAs and are highly accumulated during fetal period and neonatal
development in the brain and retina. In the present work, we studied the effect of omega-3 fatty acids in stabilization of
connections in the visual system. We tested the effect of DHA nutritional restriction or supplementation in the structure of retinal
terminal fields in the rodent superior colliculus (SC). Moreover, we tested DHA deprivation on the expression of the subunits of
AMPA and NMDA glutamate receptors and also on the expression of the phosphorylated form of GAP-43 protein (pGAP43).
Female Lister Hooded rats were fed (30g/day/animal) for five weeks before mating with either a control (soy oil) or a restricted
(coconut oil) diet. Litters were then fed until postnatal day 13 (PND13), PND28 or PND42 when they received an intraocular
injection of HRP. Another group received a single retinal lesion at the temporal periphery at PND21. In the supplemented group,
animals received fish oil daily (3g/day/animal). Omega-3 restriction induced an increase in the optical density in the superficial
layers of the SC, as a result of axonal sprouting outside the main terminal zones. In the intermediate and ventral aspects of the
stratum griseum superficiale (SGS) we observed an increased optical density of retinotectal axons during critical period
(PND13), and also at PND28 and PND42. After a contralateral retinal lesion, omega-3 restricted group showed increased
sprouting of intact axons at the lesion site as compared with the control group. Alternatively, the supplemented group showed a
focalization in fiber density in the ventral region. Furthermore, omega-3 restriction decreased pGAP-43 expression and NR1 and
NR2A subunits of NMDA receptor as well at PND28. It also increased GluR1 and GluR2 subunits expression of AMPA receptor,
suggesting a disturbance in in synaptic stability.
Conclusions:
The data indicate that omega-3 fatty acid during development impairs the topographical map organization and induces abnormal
plasticity of retinotectal axons after the closure of the critical period. This effect was correlated to increased turnover/expression
in AMPA receptors and a decreased in NMDA subunit NR1 and NR2A. These results are consistent with a role of EFA as
stabilizing synaptic agents and are essential for topographical map organization.
Keywords: Retinotectal, omega-3 fatty acids, neural plasticity, nutritional, development
Financial Support: CAPES, CNPq, FAPERJ, PRONEX, PROPPi-UFF
Resumo:22-044
ULTRA-STRUCTURAL FEATURES IN THE POSTERODORSAL MEDIAL AMYGDALA OF MALE AND FEMALE
RATS.
Ikeda, E. T. 1; Brusco, J. 2; Rasia-filho, A. A. 3; Moreira, J. E. 1,2

1
Depart. Biologia Celular e Molecular e Bioagentes Patognico, FMRP-USP
2
Depart. Neurocincias e Cincias do Comportamento, FMRP-USP
3
Depart. Fisiologia , UFSCPA
Objectives:
The rat medial amygdala MeA is involved with the processing of olfactory information and the modulation of emotions and
social behaviors. The MeA posterodorsal subnucleus MePD is implicated in sexual behavior and has receptors for estrogens and
androgens. The number of dendritic spines is different between male and female rats along the estrus cycle on the MePD. The
aim of this work is to reveal the features of the left MePD synaptic structures on male and female rats along the phases of
diestrus, estrus, and proestrus.
Two Wistar rats of each group were anesthetized and their brain fixed by transcardiac perfusion of 2% paraformaldehyde, 2%
glutaraldehyde and 0.05% CaCl in cacodylate buffer. Brain slices were post-fixed in 1% OsO 4 , and 0.4 % uranyl acetate. The
samples were infiltrated in Embed 812 resin. Ultra-thin sections 60 nm were mounted on copper single slot grids coated with
Pioloform, contrasted with uranyl acetate and lead citrate, observed and pictured under a Transmission Electron Microscope Zeiss
EM18. All the excitatory and inhibitory synapses found over dendrites, spines, soma or axons in the left MePD were
morphologically analyzed. The vesicles were classified as small round electron-lucid SRV, small pleomorfic electron-lucid
SPV, large electron-dense LEDV, and small electron-dense vesicles SEDV. Clatrin-coated and priming vesicles were
counted as well. It was used the one-way ANOVA and post-hoc test of Tukey for the statistical analyze. In the four groups
studied there is a higher number of excitatory than inhibitory synapses, with more synapses on dendrites than on spines, somas
and axons. The number of clatrin-coated vesicles in the left MePD is higher on male mean SD; 15.3 11.6 than in female rats
in diestrus 3.7 5.0, proestrus 8.6 7.8, and estrus 8.9 11.8; p < 0.01. Diestrus female have less clatrin-coated vesicles
than females in proestrus and estrus p < 0.01. The number of priming vesicles per synapse area is higher on males 6.0 5.9,
than in females in diestrus 3.1 4.8, proestrus 3.3 3.9 and estrus 4.0 5.2. The total number of vesicles per terminal,
mainly SRV and LEDV, is higher on males 369.2 137.8 than in female rats in diestrus 258 140.5, proestrus 254 125.9
and estrus 277 137.8; p < 0.01. Around 8% of synapses on dendritic spines, in the four groups studied, were
symmetricalinhibitory; they are over thin and mushroom-like spines. Multisynaptic contacts are found in 6% of the total synapses
on spines. One excitatory besides an inhibitory terminal is found in 3% of the dendritic spines.
Conclusions:
This data suggest higher activity in the left male MePD than in female, as males have more clatrin-coated vesicles, vesicles in
priming and a higher number of vesicles in general. There are descriptions of inhibitory-like terminals and multisynaptic dendritic
spines in some brain regions, but this study is the first report of these terminals in the MePD. These results provide basic
morphological data for the MePD synapses under the influence of sex hormones as neurotransmitters.
Keywords: Medial amygdala, Electron microscopy, Synapses, Estrous cycle, Sexual dimorphism
Financial Support: FAEPA, CNPQ, FAPESP
Resumo:22-045
MODULATION OF GABA RELEASE BY GLUCOSE LEVELS IN THE CHICKEN RETINA
Maggesissi, R. S. 1,2; Mya-coreixas, V. S. 2; Carpi, R. S. 2; Gardino, P. F. 1; Calaza, K. C. 2

2
Universidade Federal Fluminense, UFF
Objectives:
Recurrent hypoglycemic episodes are considered a worrying condition for insulin-treated diabetic patients health. Once diabetes
may cause vision loss, and glycemic oscillations may interfere with retinal function, this study aimed to analyze the effects of
glycemic fluctuations on morphological and neurochemical aspects of the retina.
Ex vivo chick retinas were kept under continuous perfusion with 95%O2/5%CO2 in physiological medium containing different
glucose concentrations (in mM): 0, 5.6 and 35 for 30 minutes. The consequences of hypoglycemia (0mM glucose) after 60
minutes were also analyzed. The samples were processed for Nissl staining, immunohistochemistry for -aminobutyric acid
(GABA) and lactate dehydrogenase (LDH) activity. Hypoglycemia promoted a time-dependent progressive loss of GABA
content from amacrine cells in the inner nuclear layer (INL) (meanSEM, 61%10, N=3) and ganglion cell layer (GCL) (73%6,
N=3) and its processes. This effect was due to GABA release in the first 30 minutes of hypoglycemia, since it could be blocked
by type-1 GABA transporter inhibitor (NO-711). However, a longer exposure (60 minutes) of retinas to hypoglycemic conditions
induced swelling of the inner plexiform layer (two fold), increase in LDH release (indicating cell death) (191%29, N=6) and
irreversible loss of GABA content from amacrine cells (19,6%1,5, N=3). In contrast, hyperglycemia (35mM) during 30 minutes
augmented the number of GABA-positive horizontal cells (248%21, N=3) and amacrine cells (124%9, N=6).
Conclusions:
Retinal exposure to 0mM or 35mM glucose during 30 minutes induces opposite effects on GABA release both in horizontal and
amacrine cells. Furthermore, a longer period (60 minutes) of hypoglycemia induces retinal cell death.
Keywords: GABA, Glucose, Retina
Financial Support: MCT/CNPq, INCT/INNT, CAPES, FAPERJ, PRONEX/MCT, PROPPi/UFF.
Resumo:22-046
NEUROBIOMECHANICAL OF THE RADIAL NERVE IN DIFFERENT POSITIONS OF THE UPPER LIMB.
Portes, B. B.
Centro Universitrio Nossa Senhora do Patrocnio, CEUNSP
Objectives:
Peripheral nerves of upper limb present mobility in relation to their connective tissues along its anatomic path by the neural duct
and its interfaces that are the connection for the nerves to the soft tissue and bone. Pathophysiological changes like the formation
of fibrosis connective tissue may be responsible for changes in blood flow also lead to decrease the capacity of the sliding of the
nerve into these interfaces nerve during movements of the upper limbs, thereby increasing neural tension, common in patients
with overuse syndromes. Therefore, there are tests to cause neural tension to evaluate the presence of biomechanical restriction of
the nerve when subjected to the positions of the upper limb that stress it. Thus, the objective of this job is to evaluate the
neurobiomechanical of the radial nerve in different positions of the upper limb through topography and morphometric analysis.
In this study, we used seven bodies treated with formoldehyde in which they were done topographic evaluations and metric
length analysis of the radial nerve in the following positions taken from the right upper limb: P1- Anatomic Position; P2 Abduction of the shoulder (45), internal rotation of the shoulder (45), the extension of the elbow (0) and wrist neutral (P2), P3
- shoulder abduction (45), internal rotation of the shoulder (45), extension of the elbow (0), wrist flexion (60) and ulnar
deviation of the wrist (20). The morphometric data were submitted to the ANOVA Statistic Method (p
Conclusions:
The position of upper limb, which may cause greater tension in the radial nerve associated with the movements of abduction and
internal rotation of shoulder, elbow extension, flexion and ulnar deviation of the wrist, being more suitable therefore for the use
of manual technique of the neurobiomechanical evaluation of the radial nerve.
Keywords: Morfologia, Morfometria, Nervo Radial
Financial Support: There was not support for research.
Resumo:22-047
TOPOGRAPHIC ANALYSIS OF ULNAR NERVE AT DIFFERENT POSITIONS OF THE UPPER LIMB
Arajo, J. C. G. 1; Portes, B. B. 1; Iatecola, A. 1; Cunha, M. R. 1,2

1
Centro Universitrio Nossa Senhora do Patrocnio, CEUNSP
2
Faculdade de Medicina de Jundia, FMJ
Objectives:
Aim:The neural compression syndromes peripheral upper limb are common in clinical medicine and the causes range from
trauma and occupational factors. As a result, the peripheral nerve loses its mobility along the path leading to anatomic change
sensory and motor. Technical evaluation manuals as neural tension at different positions of the upper limb as well as knowledge
of the topography of the affected nerve are essential for accurate diagnosis. Therefore, the purpose of this study was to evaluate
the topographic anatomy of the ulnar nerve in different positions of the upper limb adopted for clinical evaluation and palpation.
Methods and Results:A total of 7 formalinized cadavers were evaluated in which anatomical and measurement of the length of
the ulnar nerve as well as its distance from the middle third of the clavicle, scapula and coracoid process of the medial epicondyle
of the humerus. Measurements were performed in the following positions of the right arm: P1 - Its an anatomical position; P2 The shoulder abduction , (90), elbow flexion (90) and wrist neutral, P3 - The shoulder abduction (90), elbow flexion (90) and
wrist extension (60). The morphometric data were submitted to ANOVA (p
Conclusions:
Conclusion:The ulnar nerve has important topographical relations with bony prominences and neurovascular structures of the
upper limb with reference to the location of this nerve and understanding of associated diseases. Although no statistical difference
in the morphology between the positions adopted, we note that the position of the upper stretch increased the ulnar nerve, was
associated the movements of shoulder abduction, elbow flexion and wrist extension, so is the position most suitable for the use of
manual techniques for evaluation and mobilization of the ulnar nerve.

Keywords: Morfologia, Morfometria , Nervo Ulnar
Financial Support: Sources of research support:There was not support for research.
Resumo:23-139
BEHAVIORAL ANALYSIS OF RATS UNDER CHRONIC ANFEPRAMONE TREATMENT ASSOCIATED OR NOT
WITH EXERCISE TRAINING.
Salgueiro, J. S. 1; Borges, C. R. 1; Conceio, A. P. S. F. 1; Almeida, D. S. 1; Tieppo, C. A. 2; Lopes, C. 1,2

1
Neuroscience Nucleus/Methodist University of So Paulo, UMESP
2
Physiology /Faculty of Md. Sciences of Sta Casa de SP, FCMSCSP
Objectives:
Anfepramone, also known as diethylpropione(DEP)-(2[diethilamine]propiofenone) is one of the most used anorexigens in Brazil
for weigh control and obesity treatment. This drug acts on the central nervous system and it has the potenciality of induce
addiction and tolerance processes because of its induces mesolimbic plasticity (J. Pharmacol. Sci. Japan, 106 (1); 9-14; 2008).
This research investigate the behavioral alterations induced by the chronic treatment of female rats with anfepramone, mimicking
the main conditions of this drug use in Brazil.
Forty eight female Wistar rats, approximately 250 g and two months old were used in this research. Rats were randomized in four
groups of twelve animals, according to the 43 consecutive days received treatment: 1) anfepramone - 7 mg/kg i.p.; 2) control
anfepramone same volume of saline solution i.p.; 3) anfepramone-exercise same anfepramone treatment and 30 min of
swimming daily (water at 25oC); 4) control anfepramone-exercise saline treatment associated with 30 min of swimming daily.
Behavioral tests included three trials at 15, 30 and 43 days of experiment at the open field and at elevated plus maze with aversive
stimulus (wind and sound noise at one same closed arm). It was evaluated some parameters of general motor activity, as reagings,
groomings, stand period, excrement count on the open field and the time rats spent on the closed arm with aversive stimulus or on
the open arm. On the open field, after 43 days, anfepramone increased the grooming behavior (1.00+/-0.28 vs 3.17+/-0.55, p
Conclusions:
Chronic treatment with anfepramone: 1) increased grooming behavioral alterations in rats which is associated with stereotyped
behavior in humans; 2) exercise training induces per se the same effect; 3) decreased aversive memory in rats, which is associated
with protective behaviors in humans, exercise training promotes this memory.
Keywords: Anfepramone, Neural Plasticity, Behavioral Analysis, Exercise training
Financial Support: Neuroscience Nucleus of Health School of Methodist University of So Paulo.
Resumo:23-140
5HT1A AND 5HT1B RECEPTORS IN ZEBRAFISH ANXIETY.
Puty, B. ; Maximino, C. ; Miranda, V. ; Herculano, A. M.

Universidade Federal do Par, UFPA
Objectives:
Anxiety behaviors have been linked to the serotonergic system, especially with the 5HT1A and 5HT1B serotonin receptors. The
administration of SB-224289, an inverse agonist of the 5HT1B receptor, and WAY-100635, an antagonist of 5HT1A receptor,
allows us to evaluate the role of these receptors in a behavioral model of anxiety in zebrafish.
Adult zebrafish, with 3 months and weighting 300-500 mg, were treated acutely with SB-224289 (0.0, 2.5 or 5.0 mg/kg, i.p; n = 9
each) and WAY-100635 (0.0, 0.003 or 0.03 mg/kg, i.p.; n = 9 each). The animals were tested in the novel tank test. The
parameters analyzed were: 1- time spent on top (total time the animal spends at the top 1/3rd of the apparatus); 2- total
locomotion (number of 10 cm squares crossed) ; 3- erratic swimming (number of high-speed zig-zag swim bouts); 4- freezing
(time that the animal spends without moving). Data was presented as meanS.E.M (time on top, freezing) or medianIQR (total
movement, erratic swimming) and analyzed using parametric or non-parametric one-way ANOVAs. Both drugs produced an
inverted U-shaped effect on the time spent on the top of the tank (WAY-100635: control: 34.31 14.68, n = 7; 0.003: 164.7
36.93, n = 8; 0.03: 145 27.98, n = 9; and SB-224289: control: 14.99 5.481, n= 9; 2.5: 148.1 39.24, n= 8 ; 5.0: 101.0 18.07,
n= 8). SB-224289 showed a decrease in the number of erratic swimming (WAY-100635: control: 0 1, n=8; 0.003: 1 1, n=8;
0.03: 0 0.5; and SB-224289: control: 9.459 5.559, n=9; 2.5: 0 0, n= 8; 5.0: 0 0, n=9) . Both drugs had no effect on the
total movement (WAY-100635: control: 39.00 86.25, n=8; 0.003: 81.50 29,5, n=8; 0.03: 55 15, n=9 and SB-224289:
control:141 121, n=9; 2.5: 42 58.75, n=8; 5.0: 175 107,5, n=9).
Conclusions:
5-HT1A receptors are tonically stimulated by serotonin, leading to increased anxiety. 5-HT1B receptors are also associated with
anxiety-like behavior in zebrafish.
Keywords: receptors 5HT1A and 5HT1B, SB-224289 and WAY-100635, behavioral model of anxiety, in zebrafish
Financial Support: CNPq(Process 483336/2009-2).
Resumo:23-141
PARTICIPATION OF DOPAMINE D2-LIKE RECEPTORS IN THE HYPERLOCOMOTION EVOKED BY
NEUROPEPTIDE S IN MICE.
Souza, L. S. 1; Guerrini, R. 2; Calo, G. 3; Gavioli, E. C. 1

1
Departamento de Biofsica e Farmacologia, UFRN
2
Department of Pharmaceutical Science , UNIFE
3
Section of Pharmacology and Neuroscience Center, UNIFE
Objectives:
Neuropeptide S (NPS) is a 20 amino-acid peptide recently identified in the brain and peripheral tissues of distinct species of
vertebrates. NPS is the endogenous ligand of a G protein-coupled receptor named NPS receptor (NPSR). The NPSR is widely
expressed in the mammalian brain, and higher levels were found in the limbic system, and in neurons from dopaminergic
pathways. Various central biological actions are evoked by NPS, such as hyperlocomotion, anxiolysis, antinociception, memory
improvement and anorexic effects. Recent findings suggest that there is an interaction between dopamine and NPS systems (J
Neurochem.,115(2):475-82, 2010; Peptides, 31(5):926-31, 2010). Thus, the present study aimed to investigate the involvement of
dopaminergic D2 receptors in the mediation of psychomotor stimulant effects of NPS in mice.
Female Swiss mice (28-33 g) were housed in groups of maximum 12 per cage (33x40x17 cm) with food and water ad libitum,
and under constant temperature (232C), and a 12-h light/dark cycle (lights on at 06:00 h). The spontaneous locomotor activity
was assessed in the open-field test which was made of formica (60x60x60 cm). The distance moved (m) during 30 min was
counted by using the software Any-Maze (Stoelting, USA). In order to evaluate the behavioral effects of NPS, animals were
subjected to an esterotaxic surgery to implant a cannula into the left lateral ventricle (icv; coordinates: AP -0.6; ML +1.1; DV -1.0
mm). The effects on locomotion of the icv injection of NPS (1 and 0.1 nmol/2 l, 5 min prior to the test) and the intraperitoneal
treatment with haloperidol (0.3, 0.1 and 0.03 mg/kg, 15 min prior to the test) was assessed in this study. We observed that the
administration of NPS 1 nmol, but not 0.1 nmol, increased spontaneous locomotion in mice, particularly in the last 15 min of
observation (C=33.82.8, n=10; NPS 0.1:33.73.0, n=14; NPS 1=45.84.5*, n=8; *p
Conclusions:
These data suggest that dopamine D2-like receptors are mediating the hyperlocomotion induced by NPS in mice. Further studies
aiming to investigate the participation of dopaminergic receptors other than D2 are in progress by our research group.
Keywords: Dopamine, Locomotor activity, Mouse, Neuropeptide S
Financial Support: CNPq and UFRN/Propesq
Resumo:23-142
THE EXPOSURE OF YOUNG RATS TO NMDA ANTAGONIST OR OPIOID AGONIST PROMOTES
ALTERATIONS IN THE BEHAVIORAL RESPONSES.
Scarabelot, V. L. 1,2; Souza, A. C. 1,2; Medeiros, L. 1,2,3; Cioato, S. 1,2; D. 1,2; Fernandes, L. 4; Caumo, W. 1,2;
Torres, I. L. S. 1,2,3
1
Depto de Farmacologia Instituto de Cincias Bsicas da Sade, ICBS-UFRGS
2
Hospital de Clnicas de Porto Alegre, HCPA
3
Programa de Ps Graduao em Fisiologia - ICBS/UFRGS, PPGFISIO-ICBS/UFRGS
4
Faculdade de Frmacia - UNIVATES - Lajeado RS, UNIVATES
Objectives:
It is know that the immature nervous system is highly sensitive to the depressant effects of the anesthetic drugs and the exposure
of newborn and children to general anesthesia is a common practice on the modern medicine. For this, the objective of this study
was to evaluate the effects of acute intervention of pharmacological drugs (NMDA antagonist or opiod agonist) at P14 upon
behavioral responses evaluated at short-, medium- and long-term.
Wistar male rats at fourteen day (P14) were divided into three groups: saline (SAL), S(+)-ketamine (KET; 20 mg/kg) and
fentanyl (FEN; 0.09 mg/kg) (n=6-14). The behavioral responses were evaluated at Open-Field (OF) and Elevated Plus-Maze
(EPM) tests, and they were performed at P14 (6h after drug administration), at P30 and at P60. The behaviors evaluated at OF:
number of line crossings, fecal boluses and rearing behaviors, latency to leave the first quadrant and time of grooming; and at
EPM: number of protected head-dipping movements (PHD) and non-protected head-dipping movements (NPHD); number of
entries in the open arms (EOA) and time spent on the open arms (TOA). Data were expressed as means standard error of the
mean (SEM). One-way ANOVA was performed to compare all groups, followed by the SNK. Students t test for independent
samples was used to compare two groups (control group and FEN or KEN). Differences were considered statistically significant
if P < 0.05. The Institutional Research Committee approved all animal procedures (GPPG-HCPA: 100186), and measures were
taken to minimize pain and discomfort. At P14, when FEN e KET were compared with the SAL group, there was an increase in
inner crossings at OF (Student's t-test P < 0.05). At P30, the FEN group, at the EPM, showed a significant decrease in TOA and
EOA when compared to the SAL group (one way ANOVA/SNK, P < 0.05). At P60, the FEN group showed a significant increase
in NPHD, EOA and TOA when compared to other groups (ANOVA/SNK, P < 0.05) and to control (one way ANOVA/SNK, P <
0.05).
Conclusions:
These results indicate that fentanyl induced more changes in behavioral responses than S(+)-ketamine, especially in relation to
the anxiolytic state of rats.
Keywords: ketamine, fentanyl, young rats, open-field
Financial Support: FIPE/HCPA, CNPq, CAPES
Resumo:23-143
EFFECTS OF CANNABINOIDS IN THE PENTYLENETETRAZOLE MODEL OF CONVULSIVE SEIZURES IN
RATS
Vilela, L. R. ; Medeiros, D. ; Rezende, G. ; Oliveira, A. C. ; Moraes, M. F. ; Moreira, F. A.

Neurocincias/Universidade Federal de Minas Gerais, UFMG
Objectives:
Epilepsy is a severe neurological disorder characterized by excessive or synchronous neuronal activity, which may lead to
convulsive seizures. The substances present in the plant Cannabis sativa, as well their synthetic counterparts (termed
cannabinoids), have been proposed as a treatment for this disorder. They act mainly through the cannabinoid (CB1) receptor in
the brain, which is also activated by endogenous substances, termed endocannabinoids. However, their effects in models
predictive of anti-convulsive activity have remained controversial. Thus, the aim of this study is to test these substances against
the generalized seizures induced by pentylenetetrazole (PTZ) in rats.
Methods: Male Wistar rats (6-8/group), weighing 280-310g, received intraperitoneal injections of vehicle or the cannabinoid
WIN 55212-2 (0.3-1.0-3 mg/kg). In an independent experiment they were treated with vehicle or the CB1 receptor antagonist
AM251 (0.3-1.0-3 mg/kg). Thirty minutes later they received intravenous infusion of PTZ, at a concentration of 10 mg/mL and a
rate of 0.5 mL/min. The threshold of PTZ as well as the latency required to inducing minimum (partial myoclonus of limbs)
and maximum seizures (hyperextension of the forelimbs and hindlimbs with head flexion) were compared between groups
through analysis of variance followed by the Newman-Keuls test. Results: WIN 55212-2 (1 mg/kg) reduced both the latency (p <
0.001) and the threshold of PTZ (p < 0.01) required to induced minimal seizures, as compared to the vehicle control group.
Treatment with antagonist AM251, on the other hand (1 mg/kg), increased the threshold of PTZ (p < 0.05) required for minimum
seizure.
Conclusions:
Despite several pieces of evidence supporting anti-convulsive effects of cannabinoids, the present study points to a proconvulsive role of CB1 signaling in the PTZ model. Although the reasons for this finding are still unclear, further experiments are
under course to determine whether other interventions, such as blockade of endocannabinoid-hydrolysis, would actually attenuate
PTZ-induced seizures.
Keywords: cannabinoids , Epilepsy, convulsive seizures , endocannabinoids, pentylenetetrazole
Financial Support: Financial Support: FAPEMIG
Resumo:23-144
EARLY PRENATAL LPS EXPOSURE INCREASES SICKNESS BEHAVIOR AFTER AN IMMUNE CHALLENGE IN
ADULT RATS
Kirsten, T. B. ; Zager, A. ; Palermo-neto, J. ; Bernardi, M. M.

School of Veterinary Medicine, University of So Paulo, FMVZ-USP
Objectives:
Lipopolysaccharide (LPS) mimics infection with gram-negative bacteria and induces sickness behavior in many species generally
accompanied by a decrease in exploratory activity, social behavior, ingestive behavior, sexual behavior and induced anhedonia,
and poor learning and cognitive functions. Previous investigations by our group showed that prenatal LPS administration (100
g/kg, i.p.) on gestational day 9.5 reduces striatal dopamine and metabolite levels (3,4-dihydroxyphenylacetic acid [DOPAC] and
homovanillic acid [HVA]) in adult male rats. However, behavioral parameters linked to motor system were not changed,
including open eld general activity, haloperidol-induced catalepsy and apomorphine-induced stereotypy. The present study
investigated the prenatal LPS effects after an LPS challenge in adulthood, once adaptive plasticity processes which occur during
the animals life can mask possible behavioral changes.
For this, dams of Wistar rats were submitted to the same conditions of treatment as our previous study (LPS on GD 9.5, 100
g/kg, i.p.). The adult male pups were again treated, i.e., challenged with LPS (100 g/kg, i.p.) on postnatal day 60-67, 90 min
before the experiments. Thus, there were four groups in the experiments (n = 7-9 for each group): SAL = prenatal saline
injection; LPS = prenatal LPS injection; SAL+LPS = prenatal saline and adulthood LPS injection; and LPS+LPS = prenatal and
adulthood LPS injection. This animals were evaluated for (1) open eld general activity (traveled distance in cm and rearing
frequency); (2) spatial memory (T-maze spontaneous alternation); and (3) circulating proinflammatory cytokines interleukin-1
(IL-1) beta and tumor necrosis factor (TNF)-alpha levels. In relation to the other groups, results showed that LPS+LPS group
presented (1) decreased distance traveled (SAL = 3062.8269.73, LPS = 3085.47219.04, SAL+LPS = 2335.05180.81 and
LPS+LPS=1416.44214.75) and rearing frequency (SAL = 29.330.83, LPS = 30.001.21, SAL+LPS = 21.383.13 and
LPS+LPS=11.752.18); (2) impairment in spatial memory, i.e., hit score (SAL = 62.50% [1.0 (0.0 - 2.0)], LPS = 81.25% [2.0
(0.0 - 2.0)], SAL+LPS = 31.25% [0.0 (0.0 - 2.0)] and LPS+LPS 6.25% [0.0 (0.0 - 1.0)]); and (3) increased IL-1 beta levels
(SAL+LPS = 239.5358.10 and LPS+LPS=511.73103.45), but not TNF-alpha levels (SAL+LPS = 1021.10310.68 and
LPS+LPS=972.03163.38). In all the cases, the results were considered as significant if p < 0.05.
Conclusions:
Early prenatal LPS exposure increases sickness behavior after an immune challenge in adult rats, i.e., decreased exploratory
activity and spatial memory and increased IL-1 beta levels. Sickness behavior is a motivational state and present results might be
related with our previous results of striatal dopaminergic reduction. Taken together the data of previous and present study,
prenatal LPS modified brain plasticity, mainly of the dopaminergic system, which was masked during the animal development.
However, the immune challenge revealed these behavioral changes.
Keywords: Open field general activity test, Proinflammatory cytokines, Spatial memory T-maze
Resumo:23-145
BENEFICIAL EFFECTS OF OMEGA-3 FATTY ACIDS ON NATURALLY INDUCED ANXIETY-LIKE BEHAVIOR
BUT NOT ON MEMORY ACQUISITION IN RATS
Barcelos, R. C. S. ; Benvegn, D. M. ; Boufleur, N. ; Emanuelli, T. ; Burger, M. E.

Objectives:
To evaluate the effects of the supplementation of the fish oil (FO) on anxiety-like behavior resulting from being in a new
environment in rats.
Methods and results: Male Wistar rats (250g30) were fed with the same standard chow (SC) and supplemented for 28 days with
FO (0,1%) or soybean oil (SO- 0,1%), through of drinking water incorporation (polisorbate 80 1%). Gas chromatography (HP
6890) showed n-6/n-3 ratio (SC plus oils) of 1,31 and 10,47 for FO and SO, respectively. Rats orally supplemented with FO
(14mg/rat/day) showed lower locomotor (49,5%) and exploratory activities (48%), higher central locomotion (53%) and lower
number of fecal pellets (21%) than those treated with soybean oil (14mg/rat/day). By burying a smaller number of marbles, FOtreated rats showed reduced anxiety symptoms (56%), while their memory performance in a water-maze task was unchanged.
After 16h of fasting, FO reduced food intake (27%), confirming a less anxious behavior. Here we showed the effects of omega-3
on anxiety-like symptoms, which is related to development of stress of being in a new environment.
Conclusions:
We conclude that omega-3 can attenuate anxiety related to everyday life. Further studies should be conducted to assess the
influence of omega-3 together with anxiolytic drugs on anxiety disorders and depression.
Keywords: omega-3, fatty acids, anxiety, memory, rats
Financial Support: CNPq, CAPES, PRPGP, PROAP-UFSM
Resumo:23-146
HALOPERIDOL-LOADED POLYSORBATE-COATED POLYMERIC NANOCAPSULES SHOWED HIGHER
ANTIPSYCHOTIC EFFICACY IN RATS
Benvegn, D. M. 1; Barcelos, R. C. S. 1; Boufleur, N. 1; Pase, C. S. 1; Reckziegel, P. 1; Roversi, K. 1;

Ourique, A. F. 2; Beck, R. C. R. 2; Burger, M. E. 1
1
Departamento de e Fisiologia e Farmacologia, UFSM
2
Faculdade de Cincias Farmacuticas , UFRGS
Objectives:
Our objective was to prepare and evaluate the efficacy of haloperidol by its nanoencapsulation in polymeric nanoparticles.
Preparation and physicochemical characterization of nanocapsules suspension: Haloperidol-loaded nanocapsules (H-NC) were
prepared by interfacial deposition of preformed polymer (Int. J. Pharm. 55:r1, 1989) and a free suspension of haloperidol (0.25
mg/mL) was prepared in water using 5% (w/v) of polysorbate 80. Particle sizes, polydispersity indices and zeta potential were
measured by photon correlation spectroscopy using Zetasizer. Drug content was assayed by HPLC, using as mobile phase
methanol/potassium phosphate monobasic pH 4.0 (60:40% v/v) and detected at 254 nm. Free drug was determined in the clear
supernatant following separation of nanocapsules by a combined ultrafiltrationcentrifugation technique. Encapsulation
efficiency (%) was calculated by the difference between the total and free drug concentrations. All formulations appeared
macroscopically homogeneous similar to a milky bluish opalescent fluid (Tyndall effect). Haloperidol was efficiently
encapsulated (95 1%). Nanocapsule suspension presented particles in the sub-micrometric range (between 200 and 300 nm),
low polydispersity (60.25), negative zeta potentials, acid pH values and drug content near 100%. In vivo experiments: Acute
pseudo-psychosis was induced and maintained with amphetamine injections (AMPH-8 mg/kg- ip) each 3 hours (0, 3, 6, 9 h) in
adult male Wistar rats (n=7). After 30 min of first AMPH administration, groups designated as free haloperidol (FH),
haloperidol-loaded nanocapsules (H-NC) and blank nanocapsules (B-NC) received a single injection of each respective
formulation (0.2 mg/Kg body weight ip). The AMPH and C groups received a single injection of polysorbate 80 suspension (5%
ip). Two observers quantified the stereotyped head behavior every 15 min according to scale scores (Psychopharmacol. 102:459,
1990). KruskalWallis analysis revealed that AMPH treatment increased the head movements when compared to the control.
Rats treated with B-NC showed no reduction in head movements at any observation following each AMPH administration. On
the other hand, both H-NC and FH showed reduced behavioral scores at all observed times after the 1st and 2nd AMPH
administration, but H-NC demonstrated lower scores than FH. Following the 3rd dose, only the H-NC group showed decreased
movement scores.
Conclusions:
Our data demonstrated that the haloperidol was efficiently encapsulated and its therapeutic efficacy was improved, since it
presented a better and lasting antipsychotic effect.
Keywords: antipsychotic, haloperidol, motor effects, polymeric nanocapsule
Financial Support: CNPq, CAPES, PRPGP, PROAP-UFSM and S.S. Guterres from the School of Pharmacy
(UF
Resumo:23-147
PRE- AND POSTSYNAPTIC A2A ADENOSINE RECEPTORS CONTROL ENDOCANNABINOID SIGNALING AT
CORTICOSTRIATAL TERMINALS
Ferreira, S. G. 1,3,4; Pandolfo, P. 2; Bitencourt, R. M. 2,1; Teixeira, F. M. 1; Nunes-correia, I. 1; Takahashi, R.

N. 2; Venance, L. 3; Harkny, T. 4; Cunha, R. A. 1; Kfalvi, A. 1
1
Center for Neurosciences and Cell Biology, CNBC
2
Departament of Pharmacology, Centre of Biological Sciences, CBS-UFSC
3
Dynamic and Pathophysiology of Neuronal Networks, InsermU667, InsermU667
4
European Neuroscience Institute at Aberdeen, ENI
Objectives:
Aims: The principal relay nucleus of the basal ganglia is the striatum. Impairment of striatal synaptic plasticity is detected in
many brain disorders. Striatal synaptic plasticity is strictly modulated by adenosine and endocannabinoids. We recently found
that A2A adenosine receptors (A2AR) may control CB1 cannabinoid receptor- (CB1R)- mediated inhibition of corticostriatal
glutamate release (Martire et al., J Neurochem 116; 273; 2011). Here we aimed at further investigating the hypothesis that the
striatal adenosinergic system acts as an antagonist to the endocannabinoid system.
Methods: We mapped the co-localization of A2ARs and CB1Rs in striatal VGLUT1- and VGLUT2-positive glutamatergic
terminals with the help of fluorescent and confocal microscopy, as well as with a newly pioneered technique, immunochemical
analysis of striatal synaptosomes with flow cytometry. We performed functional assays such as displacement binding in striatal
membranes, 14C-glutamate release experiments from superfused synaptosomes, electrophysiology in brain slices, and open field
behavior. The subjects of our investigations were Wistar rats, and wild-type and A2AR and CB1R knockout mice Results:
Striatal synapses were identified by synaptophysin-positivity. 83.5 1.1% of striatal synapses were VGLUT1-positive and 67.1
1.0% were VGLUT2-positive, i.e. glutamatergic. 88.4 1.9% of striatal terminals were CB1R positive in the wild-type but not in
the CB1R knockout mouse; similarly to A2ARs; indicating a strict co-localization between the two receptors in glutamatergic
synapses. We observed that A2AR post-synaptically controlled endocannabinoid release, thereby controlling CB1R occupancy
(Bmax). Under the inhibition of diacylglycerol lipase, presynaptic A2AR inhibited CB1R binding affinity (KD). The synthetic
cannabinoid WIN55212-2 (WIN; 1 microM) inhibited the 30 mM KCl-evoked release of 14C-glutamate by 16.5 2.4% (n = 15,
p < 0.0001 by one-sample t-test) which was abolished in the presence of the CB1R antagonist, AM251 (1 microM; 98.0 5.7%
of control, n = 9, n.s.). The activation of corticostriatal CB1Rs inhibits locomotion. WIN55212-2 (3 mg/kg, ip) greatly inhibited
locomotion by 78.1 8.8% in adult male Wistar rats (n=8). The selective A2AR agonist, CGS21680 (CGS; 10 microg/kg) did
not significantly affected locomotion (-82.9 7.1% of control), but prevented the action of WIN (78.0 13.4% of control, n.s.).
Conclusions:
Conclusions: Altogether, CB1 receptor-mediated inhibition of corticostriatal terminals is strictly controlled by both pre- and postsynaptic A2AR in the same synapse, helping understand how the selective A2AR antagonist Istradefylline can fulfil its function
as a palliative medicine in Parkinsons disease.
Keywords: adenosine A2A Receptor, cannabinoid CB1 receptor, corticostriatal, glutamate , striatum
Financial Support: PTDC/SAU-OSM/105663/2008 (A.K.; F.M.T), SFRH/BD/33467/2008 (S.G.F.)
Resumo:23-148
LIPID PEROXIDATION AND SUPEROXIDE DISMUTASE ACTIVITY IN PREFRONTAL CORTEX AND
STRIATUM OF RATS AFTER NICOTINE-INDUCED KINDLING.
Gomes, P. X. L. 1; Oliveira, G. V. 1; Arajo, F. Y. R. 1; Cordeiro, R. C. 1; Nascimento, N. L. 1; Vasconcelos,

S. M. M. 1; Sousa, F. C. F. 1; Carvalho, A. F. 2; Macedo, D. S. 1
1
2
Objectives:
Experimental model of progressive epilepsy induced by electrical or chemical subthreshold stimulation is known as kindling;
however, only recently nicotine-induced kindling was pharmacologicaly and histologicaly characterized. Studies indicated that
kindling is originated in the hippocampus, although as nicotine (NIC) is an abused reinforcing agent, other brain areas may be
involved. Furthermore several lines of evidence suggest that nicotine addiction is different in women than in men. The present
study aimed to verify possible behavior differences between sex (male and female) and age (periadolescent and adult rats) in
animals submitted to the kindling, as well as, evaluate alterations in lipid peroxidation level and superoxide dismutase (SOD)
activity after NIC-induced kindling in prefrontal and striatum.
Male and Female periadolescent (PA - 35 days old, n= 8/group) and adult (AD- 60 days old, n=8/group) Wistar rats were used.
During 4 weeks, kindling seizures were induced by repeated administration of NIC 2.0 mg/kg, intraperitoneally from Monday to
Friday (based on previous dose-response curves performed in our Laboratory). Convulsant activity was evaluated according
Racine scale [2]. Thirty minutes after the last administration of NIC, animals were euthanized and prefrontal cortex (PFC) and
striatum (ST) dissected. Lipid peroxides formation was analyzed by measuring the thiobarbituric-acid reacting substances and
was determined spectrophotometrically by the absorbance at 532nm and expressed as mmol of malondialdehyde (MDA)/g tissue.
SOD activity was determined by spectrophotometry at 560nm absorbance and expressed as USOD/ g protein. For statistical
analysis ANOVA with Student Newman Keuls as post hoc was used (p< 0.05). Female PA rats were more susceptible to kindling
development as compared to male PA rats, presenting stage 5 seizures after 19 days of NIC treatment with 75% of deaths, while
male PA rats presented stage 5 seizures only after 24 days of NIC administration with 25% deaths in this group. Adult male and
female animals presented the same susceptibility to kindling development, this occurring after 17 days of treatment with no
deaths. MDA levels did not alter in the ST (Control 1.330.38; Male PA 1.961.69; Male AD 1.070.27; Female PA
1.050.21; Female AD 1.090.27). In PFC there was a significant increase in MDA content in female PA rats, female adult and
male PA and male AD animals as compared to control (Control 0.360.12; Female periadolescent 2.211.0; Male
periadolescent 0.530.09; Female adult 1.100.31; Male adult 1.910.82). SOD activity decreased in female PA, as well as,
female and male AD rats as compared to control animals in ST and PFC (ST: Control 25.411.53; Male PA 28.593.05; Male
AD 10.460.64; Female PA 8.830.36; Female AD 11.530.76; PFC: Control 19.123.58; Male PA 21.793.59; Male
AD 11.100.69; Female PA 13.901.4; Female AD 4.391.18).
Conclusions:
Changes in oxidative stress parameters in the PFC and ST are related to nicotine-induced seizures susceptibility. Comparing sex
and age variables we can notice that female PA rats are more susceptible to NIC kindling effects.
Keywords: nicotine-induced kindling, oxidative stress, sex and age
Resumo:23-149
PENTYLENETETRAZOL-INDUCED SEIZURES DECREASE NA+,K+-ATPASE ACTIVITY IN THE MICE
CEREBRAL CORTEX
Meier L. P1,1; Marquezan, B. 1; Funck, V. R. 1; Oliveira, C. V. D. 1; Mello, C. F. D. 1; Oliveira, M. S. 2

1
1Departamento de Fisiologia e Farmacologia, UFSM
2
Campus de Itaqui, UNIPAMPA
Objectives:
In the brain, Na+,K+-ATPase activity is essential for maintaining of the electrochemical gradient underlying the resting potential
and release and uptake of neurotransmitters, and a decrease in its activity alters brain excitability, depending on the degree of
inhibition and area affected. Recent studies show that mutations in the gene encoding the Na+,K+-ATPase alpha subunit cause
the appearance of seizures, and that ouabain, a Na+,K+-ATPase inhibitor, causes seizures in mice. Given this association between
Na+,K+-ATPase and seizures, and the large number of epilepsy affected individuals which are refractory to current antiepileptic
drugs available, the aim of this study was to determine whether a relationship exists between Na+,K+-ATPase activity and onset
and spreading of the seizures induced by pentylenetetrazol (PTZ), a classical convulsant agent widely used in discovery of novel
anticonvulsant drugs.
Adult male Swiss mice (3 months old, 25-30 g) were anesthetized with ketamine and xylazine and electrodes for
electroencephalographic recordings (EEG) were positioned over the parietal cortex. One week after surgery the animals were
injected with PTZ (30, 45 or 60 mg/kg, i.p.) or vehicle (NaCl 0.9% i.p.) and observed for 15 min for the appearing of clonic and
generalized seizures as well as seizure severity. After behavioral and electroencephalografical analysis animals were sacrificed
and the parietal cortex and hippocampus were removed and homogenized in Tris-HCl buffer (10 mM, pH 7.4) for determination
of Na+,K+-ATPase activity by a colorimetric method based on the rate of ATP hydrolysis in the presence and absence of
ouabain. Furthermore, the expression of the Na+,K+-ATPase alpha subunit was determined by western blot. Statistical analysis
showed that PTZ (60 mg/kg) caused convulsions in a dose-dependent manner and decreased Na+,K+-ATPase activity by 37.63 %
in the cortex [F(3,39)=2.910, P0.05 - one-way ANOVA]. By calculating the Pearson correlation coefficient we found a
significant positive correlation between Na+,K+-ATPase activity and the latency for clonic (r=0.55, P0.05 - students t test].
Conclusions:
We conclude that there is a close relationship between the decrease in Na+,K+-ATPase activity in the parietal cortex of mice and
the onset and severity of PTZ-induced seizures, and that such a decrease in Na+,K+-ATPase activity is not related to changes in
protein expression. Given the importance of the PTZ model to the development of anticonvulsants, Na+,K+-ATPase may be a
potential target for the development of new anticonvulsants.
Keywords: Na+,K+-ATPase, PENTYLENETETRAZOL, SEIZURES
Resumo:23-150
EFFECTS OF SODIUM BUTYRATE ON MITOCHONDRIAL RESPIRATORY CHAIN ACTIVITY IN AN ANIMAL

MODEL OF MANIA
Mina, F. G. 1; Steckert, A. V. 1,2; Moretti, M. 1; Valvassori, S. 1; Ronchi, N. 2; Benedet, J2; Scaini, G. 2; Dalpizzol, F. 2; Streck, E. 2; Quevedo, J. 1
1
Laboratrio de Neurocincias, UNESC
2
Laboratrio de Fisiopatologia, UNESC
Objectives:
The present study aimed to investigate the locomotor behavior and mitochondrial respiratory chain complexes in brain of rats
subjected to a model of mania induced by d-amphetamine (d-AMPH), to evaluate the effects of sodium butyrate (SB - a histone
deacetilase HDAC - inhibitor) in this context.
Adult male Wistar rats (250300 g) were obtained from the Central Animal House of Universidade do Extremo Sul Catarinense.
In protocol of reversion, rats received intraperitoneal (i.p.) injection of either d-AMPH (2 mg/kg) or saline (1 mL/kg) once a day
for 14 days. From the 8th to the 14th day, d-AMPH and saline treated animals also received saline (1 mL/kg i.p. - twice a day) or
SB (0,6 g/kg i.p. - twice a day). In prevention protocol, rats received either saline or SB (concentrations mentioned above i.p
twice a day) for a period of 14 days. From the 8th to the 14th day, animals also received saline or d-AMPH. On the 15th day of
reversion and prevention treatments, the animals received a single injection of d-AMPH or saline. The locomotor behavior was
assessed 2h after the last injection of d-AMPH or saline using the open-eld task, and mitochondrial chain activity complexes (I,
II, III and IV) was measured in brain structures (prefrontal, hippocampus, striatum and amygdala). The treatment with SB
reversed and prevented d-AMPH-induced hyperactivity. Moreover, d-AMPH decreased significantly mitochondrial respiratory
chain complexes activity in saline treated rats in prefrontal, hippocampus striatum and amygdala in both treatments, prevention
and reversion, and SB is able to prevent and reverse this impairment.
Conclusions:
The present study reinforces the need for the study with inhibitors of HDAC as possible target for new medication in the
treatment of bipolar disorder.
Keywords: d-amphetamine, mania, mitochondrial respiratory chain, sodium butyrate
Financial Support: CNPq; UNESC
Resumo:23-151
NEUROPROTECTIVE EFFECTS OF AGMATINE AGAINST BEHAVIORAL AND NEUROCHEMICAL
IMPAIRMENTS IN MICE SUBMITTED TO THE INTRANASAL MPTP MODEL OF PARKINSONS DISEASE.
Matheus, F. C. 1; Aguiar Jr. , A. S. 1; Castro, A. A. D. 1; Fiqueiredo, C. P. 1; Santos, A. R. S. D. 1; Ferreira,

J. 5; Tasca, C. I. 1; Prediger, R. D. 1
1
Farmacologia , UFSC
2
Bioqumica, UFSC
3
Centro de Neurocincias Aplicadas, UFSC
4
Cincias Fisiolgicas, UFSC
Qumica, UFSM
Objectives:
Agmatine is an endogenous substance that has been characterized as a neuromodulator. Recent studies have shown
neuroprotective effects for such polyamine in experimental models of ischemia, stress and neurodegenerative diseases. In the
present study we investigated the potential neuroprotective effects of the systemic treatment with agmatine (30 mg/kg, i.p.)
during 5 consecutive days on the behavioral and neurochemical changes induced by the intranasal (i.n.) infusion of 1-methyl-4phenyl-1,2,3,6-tetrahydropyridine (MPTP, 1 mg/nostril) in C57BL/6 aged mice (15 months old), an experimental model of
Parkinsons disease developed recently in our laboratory.
Fifteen-month-old female C57BL/6 mice were chosen since they are more susceptible to the neurotoxic effects of MPTP and they
were divided in four groups: control/control, agmatine/control, control/MPTP and agmatine/MPTP. Animals were evaluated in
activity chamber, social recognition, neurological severity score (NSS) and open field tasks, respectively, 3, 7, 13 and 19 days
after intranasal MPTP infusion to assessed the motor, cognitive, neurological and motor functions. After the behavioral
experiments, the animals were sacrificed and the brains were dissected for the evaluation of the hippocampal glutamate uptake
and imunohistochemistry for tyrosine hidroxylase (TH) in substantia nigra pars compacta (SNpc). The agmatine capacity to
inhibit monoamino oxidase B (MAO-B) activity was also evaluated in vitro. The accepted level of significance for all tests was P
< 0.05. CEUA-UFSC: PP00358. The treatment with agmatine increased about 40% the survival rate of the animals infused with
MPTP. The pretreatment with agmatine prevented the development of cognitive and the neurological impairments caused by i.n.
MPTP. The i.n. MPTP caused a decrease in the number of crossings and rearings evaluated in the open field test and the
pretreatment with agmatine was able to prevent such deficits. Agmatine did not inhibit the MAO-B activity and thus it does not
interfere directly in the limiting step in the metabolism of MPTP to MPP+. Remarkably, agmatine protected against the lost of
dopaminergic neurons in the SNpc of mice caused by i.n. MPTP. However, agmatine treatment was not able to prevent the
decrease of hippocampal glutamate uptake induced by i.n. administration of MPTP.
Conclusions:
Altogether, these results suggest that the repeated treatment with agmatine prevent, at least in part, the behavioral and
neurochemical alterations induced by i.n. MPTP administration in aged mice.
Keywords: Agmatine, Intranasal MPTP, Non-motor symptons, Parkinson`s disease
Financial Support: CNPq, CAPES, UFSC.
Resumo:23-152
INTERFERING FACTORS OF THE ROTATIONAL ACTIVITY INDUCED BY THE FOOTFAULT TEST IN AN
ANIMAL MODEL OF PARKINSONS DISEASE
Lazzaretti, C. 1,2; Borsoi, M. 1,2; Batassini, C. 1,2,3; Silva, G. R. 1; Souza, T. M. 1,2,3

1
Universidade Federal do Rio Grande do Sul, UFRGS
2
Programa de Ps-graduao em Neurocincias, PPG neuro
3
Departamento de Bioqumica UFRGS, UFRGS
Objectives:
Parkinson's disease (PD) is a progressive neurodegenerative disorder that affects 1% of the population above 55 years of age. The
experimental model of PD that uses the toxin 6-hydroxydopamine (6-OHDA) is widely used, since it causes a death of
dopaminergic neurons of the substantia nigra pars compacta. The footfault test, which is carried out in an apparatus consisted of
an elevated grid, is a tool for screening highly 6-OHDA-lesioned rats by observing whether animals present or not contextinduced ipsilateral rotational activity (CIIRA). However, it is not yet known whether CIIRA may change because of variations in
the testing apparatus, such as in its height or in placing a surrounding wall. Objectives: The aim of this study was to verify
whether altering the height of the apparatus or changing its edges may alter the frequency in which CIIRA is induced when the
footfault test is performed by 6-OHDA-lesioned animals.
Methods: Male Wistar rats (110 days old) received 6-OHDA infusions into the MFB (3 g/L, 5.5 L) and were submitted to the
footfault test 22 days later. The apparatus varied either in height (8 or 76.5 cm) or edge (presence or not of a surrounding wall 30
or 60 cm from the center). Results:It was observed that less animals presented CIIRA when placed on the apparatus with a
surrounding wall (2 out of 37) than when placed on the apparatus without it (5 out of 15; Fisher's exact test, p = 0.016). However,
there was only a trend toward this effect when discriminating the distance of the wall from the center and comparing any of these
groups with the no-wall group (30 cm: 1 out of 19; 60 cm: 1 out of 18 animals; Fisher's exact test, p= 0.066 and 0.070,
respectively). There was no difference in the frequency of CIIRA by varying the apparatus height (8 cm: 15 out of 29 animals;
76.5 cm: 18 out of 28 animals; Fisher's exact test, p =0.424).
Conclusions:
Conclusions: The presence of a surrounding wall but not the change in the apparatus height eliminates the rotational activity
induced by placing the animal on the elevated grid. These results indicate that the possibility of presenting a tigmotaxic behavior
inhibits the induction of the context-induced rotational activity, giving a hint of the underlying mechanisms involved in this
activity.
Keywords: 6-hydroxydopamine , footfault test , Parkinson disease, substantia nigra pars compacta
Financial Support: CAPES, CNPq, and Rede Instituto Brasileiro de Neurocincia (IBN-Net) # 01.06.084
Resumo:23-153
MEDIAL AMYGDALOID NUCLEUS ALPHA1 ADRENOCEPTORS MODULATES CARDIOVASCULAR
RESPONSES TO ACUTE RESTRAINT IN RATS.
Fortaleza, E. A. T. ; Scopinho, A. A. ; Corra, F. M. A.

Farmacologia. Faculdade de Medicina de Ribeiro Preto, FMRP-USP
Objectives:
Aim: Medial amygdaloid nucleus (MeA) local neurotransmission has an inhibitory influence on restraint-evoked cardiac
response, which are characterized by both elevated blood pressure (BP) and intense heart rate (HR) increases, in rats. In the
present study, we investigated the involvement of local MeA-alpha(1)-adrenoceptor on cardiovascular responses evoked by acute
restraint in rats.
Methods and Results: We used Wistar rats weighing (240-280g). It was done implanting bilateral guide cannula into the MeA to
microinjections of selective alpha(1)-adrenoceptor antagonist WB4101 or (artificial cerebrospinal fluid aCSF) vehicle. Two days
after, animals were anesthetized with tribromoethanol and a polyethylene catheter was implanted into the femoral artery for BP
and HR recording. We pretreatment the MeA with the selective alpha(1)-adrenoceptor antagonist WB4101 into the MeA (10, 15,
and 20nmoL) before the exposure of acute restraint. Acute restraint caused BP and HR increases in aCSF treated animals (n=8).
The WB4101(10nmol/100nL)-treated animals (n=5), there were no significant difference in the restraint-related MAP or HR
increases when compared with aCSF-treated animals (MAP Interaction: F5,66= 0,4711, P= 0,7964; Treatment: F1,66= 1,115,
P= 0,2949; Time: F5,66= 28,74, P < 0,0001, and HR, Interaction: F5,66= 1,024, P= 0,4109; Treatment: F1,66= 1,947,
P=0,1676; Time: F5,66= 15,43, P < 0,0001; two-way ANOVA). The WB4101(15nmol/100nL)-treated animals (n=5),
significantly reduced restraint-evoked HR increase without significant effect on the BP response, when compared with aCSFtreated animals (MAP, Interaction: F5,66= 0,5374, P= 0,7472; Treatment: F1,66= 0,8556, P= 0,3583; Time: F5,66= 19,77, P <
0,0001, and HR, Interaction: F5,66= 5,073, P= 0,0005; Treatment: F1,66= 3,558, P= 0,0637; Time: F5,66= 18,59, P < 0,0001;
two-way ANOVA). The WB4101(20nmol/100nL)-treated animals (n=7), significantly reduced restraint-evoked HR increase
without significant effect on the BP response, when compared with aCSF-treated animals (MAP, Interaction: F5,78= 0,2870, P=
0,9189; Treatment: F1,78= 0,03743, P= 0,8471; Time: F5,78= 33,02, P < 0,0001, and HR, Interaction: F5,78= 5,049, P=
0,0005; Treatment: F1,78= 12,97, P= 0,0006; Time: F5,78= 19,92, P < 0,0001; two-way ANOVA).
Conclusions:
Conclusion: Our results suggest that alpha(1)-adrenoceptor in the MeA has an facilitatory influence on restraint-related HR
responses.
Keywords: amygdaloid, adrenoceptors, cardiovascular, restraint
Financial Support: Financial Support: CNPq, CAPES and FAEPA.
Resumo:23-154
EVIDENCE OF THE ROLE OF SPINAL C-JUN-NH2-TERMINAL KINASE (JNK) ACTIVATION IN THE
DEVELOPMENT OF SPINAL CORD INJURY.
Martini, A. C. ; Forner, S. ; Koepp, J. ; Rae, G. A.

Dept. of Pharmacology/Federal University of Santa Catarina, UFSC
Objectives:
Traumatic spinal cord injury (SCI) is a complex devastating neurological disorder that compromises major motor, autonomic and
reflex functions, leading to permanent homeostatic disabilities. SCI affects 2.5 million people worldwide (~ 250,000 in Brazil)
and remains an important cause of mortality (Nat. Rev. Neurosci., 7: 628, 2006). c-Jun-NH2-terminal kinase (JNK) is a stressactivated MAPK implicated in many aspects of cellular regulation including proliferation, gene expression and programmed cell
death (Bioch. Bioph. Acta, 1773:1341, 2007). The present study assesses changes in spinal levels of MPO and apoptotic cells,
and the potential role of spinal JNK activity in promoting the motor impairment inflicted by the lesion.
A moderate degree of paraplegia was induced in anesthetized male Wistar rats (270-300 g; 4 6 per group) by inserting a Fogarty
2F embolectomy catheter (3 mm diameter, 5 mm length) at the T10 level and inflating it for 1 min (J. Neurotrauma, 18: 1399,
2001). At 2, 6, 24 and 72 h after SCI or sham surgery, spinal cord segments (T8 to T12; ~ 2 cm long) were collected for Western
blot analysis of expression of total and phosphorylated forms of JNK protein, myeloperoxidase (MPO) activity assays and
TUNEL analysis of apoptotic cells. Rats were treated intrathecally twice with SP600125 (a JNK inhibitor; 150 nmol) or vehicle at
1 and 4 h after SCI. Recovery from motor disturbance was graded every other day, for 4 min, during 28 days, using a locomotor
deficit scoring system (BBB scale) (J. Neurotrauma, 12: 1, 1995). All protocols were approved by UFSCs Committee on the
Ethical Use of Animals (23080.008708/2010-41). Relative to sham-group values, expression level of total JNK MAPK didnt
change after surgery, while that of activated JNK rose by 467,84 86,60 % and 654,65 82,71 % at 6 and 24 h, respectively.
Spinal cord MPO levels at 6, 24 and 72 h after SCI increased by 417 95, 437 82 and 233 40%, respectively. Almost no
apoptotic cells were detected in the spinal cord from sham-operated rats, while SCI animals presented 2,62 0,76 and 8,54 2,07
TUNEL positive-cells per field at 24 and 72 hours, respectively. The JNK inhibitor treatment reduced the expression level of
phospho-JNK (70,18 6,00%) and the number of apoptotic cells (71 11%) at 6 hours after SCI, without changing MPO levels.
More importantly, SCI rats treated with SP00125 displayed higher BBB scores than the vehicle-treated SCI controls as from 8th
day after SCI onwards (BBB scores at 2, 8, 14 and 28 days after SCI: SP600125 group 0.64 0.42, 6.36 1.21, 9.36 1.24; 12.5
1.88; vehicle group 0.0 0.0, 2.83 0.95, 5.92 0.89; 6.8 0.93).
Conclusions:
These results show a close association between activation of spinal JNK and initiation of inflammatory responses and apoptosis
after SCI. Moreover, they suggest that inhibition of JNK activation in the spinal cord might hold therapeutic potential for
functional recovery from SCI.
Keywords: c-jun-NH2-terminal kinase, locomotor recovery, spinal cord injury
Financial Support: CNPq, FAPESC, PRONEX, CAPES.
Resumo:23-155
RIMONABANT REPEATED TREATMENT INHIBITS C-FOS EXPRESSION IN COCAINE-SENSITIZED MICE
Yokoyama, T. S. 1,2; Marinho, E. A. V. 1,2,3; Oliveira-lima, A. J. 1,3; Santos, R. 1; Hollais, A. W. 1; Ribeiro,

L. T. C. 1; Estebanez, D. N. 1,2; Ruiz, A. R. 1,3; Longo, B. M. 4; Frussa-filho, R. 1
2
REA DA SADE, UBC
1
FARMACOLOGIA, UNIFESP
3
REA DA SADE, FIT
4
NEUROFISIOLOGIA, UNIFESP
Objectives:
: Drug addiction is a chronically relapsing disorder characterized by neuroplasticity and neuroadaptations in different brain areas
such as ventral tegmental area (VTA), nucleus accumbens (Nacc), dorsal striatum (DS), prefrontal cortex (PFC) and amygdale.
Drugs of abuse produce, in rodents, locomotor stimulation, a behavior that is sensitized after repeated administration. This
behavior sensitization phenomenon is proposed to share neuronal mechanisms with drug craving. We investigate the expression
of the c-fos protein in brain areas of the CNS in female mice (25-35g) previously and repeatedly treated with cocaine and
subsequently treated with the cannabinoid antagonist rimonabant
Swiss mice received 10mg/kg cocaine or saline every other day for 15 days. From the 17th to the 24th day the animals were daily
treated with vehicle or rimonabant (1 or 10mg/kg). On the 30th day all the animals were challenged with 10mg/kg cocaine and 1h
after were anesthetized and perfused with paraformaldehyde. The brains were removed and cryoprotect in sucrose phosphate
buffer. Coronal 30 m thick sections were collected for c-fos immunohistochemistry in the basolateral amygdala (BLA), PFC,
DS, Nacc and VTA. The following groups were formed: Saline-Vehicle-Cocaine; Saline-Rimonabant1-Cocaine; SalineRimonabant10-Cocaine; Cocaine-Vehicle-Cocaine; Cocaine-Rimonabant1-Cocaine; Cocaine-Rimonabant10-Cocaine. Cocaine
repeated administration produced locomotor sensitization and its expression was attenuated by the subsequent treatment with
rimonabant. The expression of c-fos was increased in the Cocaine-Vehicle-Cocaine group in BLA (20.50.86), PFC (46.71.2)
and DS (29.53.7) when compared to Saline-Vehicle-Cocaine group (BLA=9.81.3), (PFC=24.73.9), (DS=14.72.8) (p
Conclusions:
Our results indicate that repeated treatment with cocaine promoted the activation of different, but related, structures involved with
drug dependence. It was also observed that subsequent treatment with rimonabant was effective at reversing some of cocaineinduced neuroadaptations
Keywords: BEHAVIORAL SENSITIZION, C-FOS EXPRESSION, RIMONABANT
Financial Support: FAPESP, AFIP and CNPQ.
Resumo:23-156
SUITABILITY OF FEMALE MICE IN THE STUDY OF ALZHEIMERS DISEASE.
Monteiro-silva, K. C. 1; Nunes, M. A. 1; Schwe, N. M. 2; Caetano, A. L. 1; Vieira, L. M. S. 1; Telles, M. 3;

Viel,t. A. 2,3; Buck, H. S. 1
1
Department of Physiological Sciences, FCMSCSP, FCMSCSP
2
Institute of Biomedical Sciences, USP, USP
3
School of Arts, Sciences and Humanities, USP, USP
Objectives:
Contemporary ethical procedures encourage the refinement, reduction or replacement of animals in biological research.
Transgenic animals are expensive and difficult to obtain, so the use of female transgenic mice in the study of Alzheimer disease,
as well as in other studies, provides economy of funds, better utilization of animals and, most important, being in accordance with
the ethical procedures. Based on that, this study evaluated the suitability of female mice in the study of cognitive disorders in
Alzheimers disease
METHODS: Six-months-old transgenic mice TG(PDGFB-APPSwInd) male (mTG, n=11) and female (fTG, n=5), expressing
human amyloid precursor protein and their age-related wild type (C57Bl/6) male (mWT, n=8) and female (fWT, n=10) were
used. The spatial memory was evaluated at Barnes maze (5 blocks of two trials), where the decreases of latency time to find an
escape box indicates learning had occurred. Memory was also evaluated by inhibitory avoidance test (0,5 mA, 2 sec, max latency
of 300 sec), which is associated to an aversive stimulus. The obsessive-compulsive behavior was evaluated by the marble test,
where the number of buried marbles and the time spent digging the ground was counted. The anxiety was evaluated by the
elevated plus maze, where the number of entries in the open arms, in the closed arms and head dipping were recorded. The motor
activity was evaluated using an activity cage. The Data were analyzed by Two-way ANOVA or Students T test and where
different when P< 0,0001) but not in factor Gender (F(1,50)=0,64; p=0,43), similar data were observed in WT mice
(Time=F(2,50)= 28,48; p
Conclusions:
Female transgenic mice PDGFB-APPSwInd could be useful to study cognitive changes in Alzheimers disease as well as their
wild type background C57Bl/6.
Keywords: Alzheimer's disease, behavior, female, memory
Resumo:23-157
ANTS DEFENSIVE BEHAVIORS ARE MODULATED BY THE NEONICOTINIOID IMIDACLOPRID
Galvanho, J. P. 1; Samuels, R. I. 1; Carrera, M. P. 1

Lab de Entomolofia e Fitopatologia/U. E. do Norte Fluminense, UENF
1
Lab. de Sanidade Animal/Univer. Estadual do Norte Fluminense, UENF
2
Objectives:
Leaf-cutting ants are insects of economic interest because they are pests of various types of plantations. A possible tool for the
control of these insects could be the combination of sub-lethal doses of imidacloprid (IMI) with spores of entomopathogenic
fungi. Therefore, this study aimed to determine the effects of IMI and the entomopathogenic fungus Beauveria bassiana on the
locomotion and self-grooming behavior of the ants [experiment (exp) 1], and also on the agonistic behaviors, allogrooming and
antennal contact of ants in a group situation [exp2].
The major worker caste of A. subterraneus subterraneus (HYMENOPTERA:FORMICIDAE) were treated with IMI at 10, 20 and
40 ng/insect and conidial suspensions of the fungus at a concentration of 1x108 conidia/ml. IMI was topically administered to the
thorax and the fungus was administered by contact of the ants with a filter paper impregnated with a conidial suspension. All ants
were acclimatized for one hour before receiving their respective treatments that lasted four hours. In exp 1 the treatments were:
IMI10 (10 ng/kg) IMI20, IMI40, fungal suspension (F), IMI10+F; IMI20 + F and IMI40 + F. In exp 2, one ant was treated with
fungus and then placed together with 10 control ants (forming the group 10CONT+1F) or with 10 ants treated with IMI10
(forming the group 10IMI+1F). The behaviors were recorded and processed by ANOVA for repeated measurements and the
differences were verified using Duncans test. For exp1, results showed that groups with statistical significance were IMI10,
IMI10 + F and F. The IMI10 group showed higher locomotor activity (1385,5cm+91,5) and reduced self-grooming (2,9s+0,3)
when compared to control (LA:974,6cm+53,5; SG:18,1s+1,8) and the IMI20 (LA:1098,2cm+61,3; SG: 18,4s+4,1) and IMI40
(LA: 876,4cm+86,2; SG: 16,9s+4,6) groups. The same was observed for IMI10 + F (LA:1093,0cm+51,2; SG:6,0s+0,6), however
when compared to control and IMI20+ F (LA:727,4cm+110,8; SG:25,5s+5,4) or IMI40 + F (LA:577,9cm+41,3; SG:25,5s+ 5,4)
groups, the F group showed lower locomotor activity (902,5cm+107,6) and exacerbation of self-grooming (87,8s+7,8) compared
to controls. For exp2, results showed that the presence of ants treated with F inhibited the agonistic actions (1,0 score+0,2) and
exacerbated allogrooming (9,5s+2,0) and antennal contact (39,6s+3,3) of control ants to the fungus treated ant [10CONT+1F].
However, changing the experimental design from 10 CONT to 10IMI [10IMI+1F group] resulted in reversal of behaviors, in
other words, exacerbation of aggression (8,3 score+0,8) and a reduction of allogrooming (0,1s+0,1) and antennal contact time
(16,4s+1,1) of the ants exposed to the fungus placed together with 10 ants treated with IMI (10IMI+1F). Therefore, IMI blocked
the behaviors produced by fungus in individually observed ants and changed the social interaction pattern of infected ants,
observed in a group situation.
Conclusions:
Thus, nicotinic system modulation by IMI changes defensive behaviors in both individual and social situations. This suggests that
IMI could be a useful tool in the control of leaf-cutting ants.

Keywords: Defensive Behaviors , Acromyrmex subterraneus subterraneus, Imidacloprid, Beauveria bassiana, Biological control
Financial Support: UENF
Resumo:23-158
MEDIAL PREFRONTAL CORTEX CHOLINERGIC NEUROTRANSMISSION MODULATES THE EXPRESSION
OF CONTEXTUAL FEAR CONDITIONING IN RATS.
Fedoce, A. D. G. ; Junior, N. C. F. ; Reis, D. G. . D. ; Correa, F. M. A. ; Resstel, L. R. M.

Departamento de Farmacologia faculdade de Medicina , FMRP-USP
Objectives:
Introduction: The ventral portion of medial prefrontal cortex (vMPFC) is involved with modulation of behavioral and autonomic
responses evoked by stress situations. There is evidence showing that vMPFC local cholinergic system is able to modulate
aversive responses. Moreover, during expression of fear conditioning the vMPFC acetylcholine levels is increased. Thus, our
work investigated the involvement of cholinergic system present in vMPFC on behavioral (freezing) and autonomic, increase of
mean arterial pressure (MAP), heart rate (HR) and decrease of tail temperature responses, of rats re-exposed to a conditioned
aversive context.
Methods: Male Wistar rats (230-270g) had guide cannulae bilaterally implanted in the vMPFC. Twenty four hours before the test
session, animals were submitted to conditioning session (6 foot electrical shock, 1.5 mA, 3 s or 3 foot electrical shock, 0.8 mA,
2s).Acatheter was implanted in the femoral artery for cardiovascular recordings. Hemicholinium (1nmol), which decreases the
synthesis of acetylcholine, was administered 20 min before the chamber re-exposition (test session). Neostigmine (1nmol),
reversible acetylcholinesterase inhibitor, was administered 10 min before the chamber re-exposition (test session). Muscarinic
antagonist atropine (6nmol), muscarinic M1 and M3 antagonist j104129 fumarate (6nmol) were administered 10 min before the
test session. Time spent in freezing and cardiovascular responses were recorded for 10 min. Results: The hemicholinium (n=8)
reduced the freezing (8 1.5 % vs 69 8 %; t= 7.0, P<0.001).
Conclusions:
Conclusion: Our findings show that vMPFC cholinergic neurotransmission, through M1 and M3 receptors, modulates the
expression of behavioral and autonomic responses evoked by contextual fear conditioning. Financial support: FAPESP, CNPq
and FAEPA.
Keywords: cholinergic, contextual fear conditioning, Medial prefrontal cortex, neurotransmission
Resumo:23-159
INVOLVEMENT OF DOPAMINERGIC MECHANISMS IN THE EXPRESSION OF CONTEXTUAL CONDITIONED

FEAR IN RATS.
Caetano, K. A. S. ; Oliveira, A. R. ; Brando, M. L.

Universidade de So Paulo - FFCLRP - Depart. de Psicologia, USP
Objectives:
It is well established that threatening situations that generate fear reactions are practically unforgettable. In this way, aversive
conditioning induces several defensive behaviors, like freezing, a signal of fear in rodents, being this response characterized by
the absence of all observable movements, except those related to respiration. The knowledge of the behavioral and physiological
components involved in these defensive responses can be useful for the understanding and establishment of effective therapeutics
for anxiety disorders. It is known that dopamine is an important neurotransmitter of mammalians brain, mainly by modulating
the behaviors related to reward. On the other hand, in recent years many researchers have noticed an enhancement of
dopaminergic neurotransmission in structures of the mesocorticolimbic system after exposure to stressful events. However,
conflicting results obtained in several animal models of anxiety urge the need of further investigation about dopaminergic
modulation of aversive states. Thus, the aim of the present work was to evaluate the involvement of dopaminergic
neurotransmission in the expression of contextual conditioned fear in rats.
Methods: 74 male Wistar rats weighting 250-300 g were subjected to contextual conditioning (10 footshocks, 0.6 A). In the test
session, conducted 24 hours after the conditioning session, the animals received intraperitoneal injections of D1 and D2 agonists
(SKF 38393 and quinpirole) and antagonists (SCH 23390 and sulpiride) and tested during 10 minutes in the same box where
conditioning occurred. The time of freezing behavior was measured during this session. The motor activity was evaluated in the
open field test. Results: The administration of the D2 agonist (quinpirole) and antagonist (sulpiride) decreased the contextual
conditioned freezing [F(2,23) = 44,58; p < 0,001 for quinpirole; F(3,29) = 4,86; p < 0,05 for sulpiride]. The injection of the
D1 agonist (SKF 38393) and antagonist (SCH 23390) did not affect the conditioned freezing. Motor activity was not affected by
these drugs.
Conclusions:
The D2, but not D1, receptors are involved in the expression of contextual conditioned fear. It is possible that quinpirole and
sulpiride acted on presynaptic and postsynaptic D2 receptors, respectively, leading to a decrease of dopaminergic
neurotransmission in both cases.
Keywords: Conditioned fear, Dopaminergic neurotransmission, Anxiety
Resumo:23-160
BLOCKADE OF THE ANXIOLYTIC-LIKE EFFECT INDUCED BY WAY100635 MICROINFUSION INTO THE
MEDIAN RAPHE NUCLEUS BY PRIOR KETANSERIN INJECTION INTO THE PERIAQUEDUCTAL GREY
MATTER IN MICE EXPOSED TO THE ELEVATED PLUS-MAZE.
Nunes-de-souza, V. 1; Rodgers, R. J. 3; Nunes-de-souza, R. L. 2; Canto-de-souza, A. . 1

3
University of Leeds, UL
1
Universidade Federal de So carlos, UFSCar
Universidade Estadual Paulista, UNESP Campus Araraqu
Objectives:
A wealth of evidence supports the involvement of the serotonergic neurons of the median raphe nucleus (MRN) in anxiety. It has
been known that the blockade of 5-HT1A somatodendritic autoreceptors in the midbrain raphe nuclei increases the firing rate of
these neurons, disinhibiting 5-HT release in postsynaptic areas such as amydala, hippocampus and periaqueductal grey matter
(PAG). Whereas stimulation of 5-HT1A or 5-HT2 receptors in amygdala and hippocampus enhances anxiety-like behaviors in
rodents, activation of both receptor subtypes in the PAG markedly reduces anxiety-like behavior. The present study investigated
whether the anti-anxiety effects induced by MRN 5-HT neurons desinhibition (Brain Res. 928: 50, 2002) are attenuated by the
blockade of 5-HT2 receptors located within the PAG.
Mice received combined intra-PAG injection with ketanserin (0 or 10 nmol/0.1 l), followed by intra-MRN injection of WAY100635 (0 or 5.6 nmol/0.1 l) ten minutes later. They were individually exposed to the elevated plus-maze (EPM) for five
minutes. The results showed that intra-MRN WAY100635 injection reduced behavioural indices of anxiety without significantly
altering general activity measures. Thus, microinjection of WAY100635 increased percentage of open arm entries [%OE - V + V:
42.9 3.6; V + WAY: 54.4 3.7, P < 0.05] and percent open arm time [%OT - V + V: 31.2 1.2; V + WAY: 48.7 4.1, P <
0.05]. This anti-anxiety effect of WAY100635 was blocked by prior injection with an intrinsically inactive dose of ketanserin
(Ket) in the PAG [%OE: Ket + WAY: 44.9 1.4; %OT: Ket + WAY: 31.5 1.7]. Neither WAY nor Ket changed closed arm
entries.
Conclusions:
These data suggest that 5HT2 receptor populations located within the midbrain PAG played a role in the anxiolytic-like effect
induced by MRN 5-HT neurons disinhibition.
Keywords: anxiety, 5-HT, Elevated Plus Maze, mice
Financial Support: UFScar, CNPq
Resumo:23-161
BEHAVIORAL AND MORPHOLOGICAL CHANGES IN THE NEMATODE CAENORHABDITIS ELEGANS
UNDER ZIPRASIDONE TREATMENT.
Gubert, P. ; Mourao, T. ; Aguiar, G. C. ; Bridi, J. ; Barros, A. G. D. A. ; Romano-silva, M. A.

Departamento de Sade Mental, UFMG
Objectives:
Second generation antipsychotics (SGA) are used for treatment of psychiatric disorders such as schizophrenia. It is wellestablished the relation between SGA and metabolic side effects, even though this is a variable outcome among them (Psychiatr.
Serv. 59:515, 2008). Compared to others SGAs, ziprasidone (ZIP) has a similar pharmacodynamic profile, however, a relatively
low risk for weight gain or others metabolic shifts. The mechanisms underpinning it remain obscure. The nematode
Caenorhabditis elegans is a genetic tractable model suitable for depict molecular pathways behind behaviors (Nat. Rev. Drug
Discov. 5:387, 2006). Recently, studies have reported that olanzapine and clozapine modulate size and growth on Caenorhabditis
elegans through mechanisms not involving serotonin or dopamine (Pharmacol. Res. 54:361, 2006). The goal of the present study
was to determine the impact of ZIP treatment on the nematode behaviors.
Worms were cultured under standard conditions on E. coli OP50 bacterial lawns (Genetics 77:71, 1974). N2 bristol was used as
wild type reference strain. Ziprasidone was diluted in DMSO and added to plates filled with NGM and bacteria to a final
concentration of 40 M. L4-synchronized nematodes were exposed to the drug or mock condition and assessed for the desired
parameters after 24 h. The pumping assay was performed by counting the number of pharyngeal pumps in 30 s, while for the
defecation assay, the interval between defecation cycles were gauged. Exposure to ziprasidone did not changed defecation or
pharyngeal pumping rates. We also analyzed worms movement and length measuring their velocity, acceleration, distance and
body size. A statistically significant decrease in distance crossed (p < 0.05, Student's t-test, n=20) and body length (p < 0,001,
Student's t-test, n=20) were observed in ZIP (40M) treated animals compared to control (DMSO). Other interesting effect was
observed after counting the number of eggs inside worms uterus. Exposure to ZIP caused a reduction in egg production (p <
0.0001, Student's t-test, n=20).
Conclusions:
The present study revealed some interesting effects of exposure to ziprasidone in C. elegans behavior and morphology. Previous
studies have shown pathways and neurons engaged in the control of the tested behaviors. Therefore it is possible that the same
actors may play a role in ZIP exposure outcomes. Our findings call for further studies to elucidate the molecular mechanisms
implicated in changes induced by ziprasidone in the nematode and, consequently, shed light into fresh mechanisms that can
justify aspects of ZIP use in humans.
Keywords: antipsychotic, behavior, Caenorhabditis elegans, ziprasidone
Financial Support: CAPES, CNPQ, FAPEMIG, INCT-MM.
Resumo:23-162
INTERACTION BETWEEN MUSCARINIC AND 5-HT3 RECEPTORS IN THE MEDIAL SEPTAL AREA ON
CARDIOVASCULAR CONTROL IN RATS.
Batista, A. D. S. 1; Nascimento, N. C. S. 1; Santana, R. C. 1; Ferreira, H. S. 2; Fregoneze, J. B. 1

1
Dep. Bio-Regulao/Universidade Federal da Bahia, UFBA
2
Dep. Cincias da Vida/ Universidade do Estado da Bahia, UNEB
Objectives:
The medial septal area (MSA) is a limbic structure that receives serotoninergic afferents from the raphe nuclei and presents high
expression of cholinergic receptors. It has been demonstrated that brain serotoninergic pathways are involved in the control of
blood pressure. Previous data from our laboratory show that the activation of 5-HT3 receptors in the MSA decreases the stressinduced hypertensive response. Furthermore, the blocked of these receptors in non-stressed rats evokes an increased in blood
pressure. The aim of the present study was to investigate the possible interaction between muscarinic and 5-HT3 receptors in the
MSA on the control of blood pressure.
Male Wistar rats (280-310g) were anaesthetized with Ketamine/xylazine (80/11.5 mg/kg i.p) for guide cannula implant in the
MSA. The animals were anaesthetized again, 24 h before the experiment, for carotid artery catheterization. At the beginning of
the experimental session the carotid catheter was connected to a blood pressure transducer and this to a computer system for data
acquisition. The baseline blood pressure was recorded for 30 min. Afterward the rats were divided in the following groups
receiving two consecutive microinjections into MSA 10 min apart from each other: 1) atropine (5 nmol), a muscarinic cholinergic
antagonist plus ondansetron (160 nmol), a 5-HT3 receptors antagonist (ATRP+OND); 2) atropine plus saline (ATRP+SAL); 3)
saline plus ondansetron (SAL+OND); and 4) saline plus saline (SAL+SAL). At the end of the experiments the animals were
anesthetized, submitted to a transcardiac perfusions with saline followed by 10% formalin and have their brain removed for
histological procedure. Only data from animals whose guide cannulas were in the MSA were considered. The data are expressed
as delta of mean arterial pressure (MAP) and heart rate (HR), calculated from the basal values. The statistic analysis used was
two-way ANOVA for repeated measure followed by the post-hoc Bonferroni test (p< 0.05). The results show that the
pretreatment with atropine reduces the hypertensive response induced by 5-HT3 receptors blockade in MSA. The most intensive
inhibition was observed at 35 min after atropine microinjections into MSA (ATRP+OND delta PAM= 2.7 1.0 mmHg, (n=4)
and SAL+OND delta PAM= 24.4 3.1 mmHg (n=4). The heart rate did not change with any of the treatment.
Conclusions:
The results of this study show that the hypertensive responses evoked by the blockade of 5-HT3 receptors in the MSA are
dependent of muscarinic receptors activation. The data suggest that the interaction between the muscarinic and 5-HT3 receptors
in the MSA is important for cardiovascular regulation in rats.
Keywords: 5-HT3 receptor, Muscarinic receptor, Blood pressure, Medial septal area
Financial Support: CAPES, CNPq e FAPESB.
Resumo:23-163
CAENORHABDITIS ELEGANS AS AN EXPERIMENTAL MODEL FOR ANTIPSYCHOTIC BEHAVIOUR AND
DEVELOPMENTAL EVALUATIONS
Mouro, T. ; Gubert, P. ; Aguiar, G. C. ; Bridi, J. ; Barros, A. G. D. A. ; Romano-silva, M. A.

Departamento de Sade Mental / Faculdade de Medicina, UFMG
Objectives:
Caenorhabditis elegans is a free-living nematode used as an experimental model due to its short lifetime and wide range of
available genetic tools. Moreover, its transparent body, constancy of cell number and position and simple nervous system (302
neurons in hermaphrodites) has been considered unique advantages to the study of development and neuropharmacology. These
features offer the possibility to identify novel molecular targets related to mental disorders and compounds employed in
therapeutics. Recently, the model was used to assess the adverse effects of some second-generation antipsychotics (SGAs)
(Pharmacol. Res. 54:361, 2006). Ziprasidone (ZIP) has fewer adverse effects, especially those related to weight gain, compared to
others SGAs. Therefore, we decided to evaluate the effects of ZIP exposure in C. elegans. We measured the outcome of longterm exposure to ZIP in the nematode length, development and movement.
Worms were cultured under standard conditions on E. coli OP50 bacterial lawns (Genetics 77:71, 1974). N2 bristol was used as
wild type reference strain. Ziprasidone was diluted in DMSO and added to plates filled with NGM and bacteria to a final
concentration of 40 M. For developmental evaluation, we assessed worms size and sexual development. Our findings pointed
that ziprasidone did not affect C. elegans development which was similar to what has been showed to other atypical
antipsychotics (Pharmacol. Res. 54:361, 2006). Conversely, ZIP exposure reduced the nematode progression through larval
stages and caused an unusual dispersion of worms on plates.
Conclusions:
Ziprasidone did not affect the development of Caenorhabditis elegans in a long-term exposure paradigm. Nevertheless, ZIP
changed the body length and dispersion of worms. Therefore, we believe that ZIP modulates molecular pathways related with
development progression. Further analysis will be important to specify the pathways and shed light into possible new targets of
the compound.
Keywords: Antipsychotic, Caenorhabditis elegans, Development, Ziprasidone
Financial Support: CAPES, CNPQ, FAPEMIG, INCT-MM
Resumo:23-164
EFFECTS OF PRETREATMENT WITH HIGH VS. LOW DOSES OF TYPICAL ANTI-PSYCHOTIC,
HALOPERIDOL AND ATYPICAL OLANZAPINE ON THE BEHAVIORAL SENSITIZATION INDUCED BY
APOMORPHINE
Dias, F. R. C. 1; Matos, L. W. . D. 1; Carrera, M. P. 1; Carey, R. J. 2

Universidade Estadual do Norte Fluminense Darcy Ribeiro, UENF
2
VA MEDICAL CENTRER , VA MEDICAL CENTRER
Objectives:
The anti-psychotic drugs are potent antagonist at the dopamine D2. The apomorphine is a mixed D1/D2 agonist that has been
repeatedly and reliably demonstrated to induce potent locomotor sensitization. Repeated administration the dopamine agonist and
antagonist produce different changes in dopamine receptor sensitivity. The antagonist produces increases in post-synaptic
receptor sensitivity, in contrast, dopamine agonist desensitize the inhibitory autoreceptors. The aim of this study was to
investigate the effects of pretreatment with haloperidol (HAL) and olanzapine (OLA) at doses of 1.0 mg/kg (acts blocking the
postsynaptic receptors) and at the doses of 0.01 mg/kg (HAL) and 0.03 mg/kg (OLA) (acts blocking the presynaptic receptors) on
the sensitization induced by apomorphine (APO - 2.0 mg/kg).
For this, in two experiments, separate groups of rats were administered ten daily low or high doses of either HAL or OLA, which
was the supersensitivity phase (SP) and locomotion was recorded for 40 minutes. After five days of withdrawal, all groups were
given injections of APO on 5 successive days, which was the induction phase (IP). The control group received vehicle. After a
withdrawal period, all animals were administered with APO, which was the sensitization expression (SE). The control group
received vehicle. The results showed that the SP, both the high dose HAL and OLA groups exhibited a profound suppression of
locomotion activity. While neither the low dose HAL or OLA treatments initially had an effect upon locomotion activity, low
dose HAL, but not OLA, increased locomotion activity with repeated treatments both the high dose HAL and OLA groups
exhibited a profound suppression of locomotion activity. In the IP, of the experiment, the APO treatments given to the vehicle
control group generated a typical progressive locomotion sensitization effect. This sensitization effect was not modified by either
the low or high dose OLA treatments. In contrast, the low and high dose HAL treatments differentially modified the apomorphine
sensitization effect. Low dose HAL potentiated apomorphine sensitization; whereas, high dose HAL initially potentiated but
subsequently attenuated APO sensitization.
Conclusions:
Atypical anti-psychotic drugs can protect against the development of dopamine receptor supersensitivity. Although repeated
typical anti-psychotic treatments can induce dopamine receptor supersensitivity, this effect can be reversed by repeated dopamine
agonist treatments. Paradoxically, repeated low dose haloperidol treatment can function as a dopamine agonist.
Keywords: Haloperidol, Olanzapine, Behavioral sensitization, Apomorphine
Financial Support: FAPEJ/CNPQ
Resumo:23-165
EVALUATION OF PARAMETERS OF OXIDATIVE STRESS IN STRIATUM OF ADULT MALE AND FEMALE
RATS THAT UNDERWENT NEONATAL HANDLING
Arcego, D. M. 1; Noschang, C. 1; Krolow, R. 1; Quillfeldt, J. A. 2; Dalmaz, C. 1

1
Departamento de Bioquimica/ICBS, UFRGS
2
Instituto de Biociencias, UFRGS
Objectives:
To evaluate whether neonatal handling affects oxidative stress parameters in the striatum in adult male and female rats
Methods:16 pregnant Wistar rats were used. The day of birth was considered day 0, when it was standardized at 8 pups per litter
and litters divided into non-handled and handled. The animals were subjected to a cycle of normal 12 hours light / dark, lights on
at 7:00 h, with standard chow and water ad libitum. Rats in the handled group were placed in an incubator (32C) for 10
min/day, days 1-10 after birth. The puppies were separated by sex at 21st postnatal day. At 60 days of age, the animals were
sacrificed and striatum was removed and frozen at -70C until analysis of enzymatic activities of Superoxide Dismutase (SOD),
glutathione peroxidase (GPx), Catalase (CAT), the production of free radicals by chemical oxidation of diclorofluorescena
(DCF) and determination of total thiol content. Just one male and one female were used per litter for oxidative stress study. The
results were analyzed by Students t test. Results: Results are shown as mean + S.E.M. of U/mg protein (SOD); mol H2O2
transformed/min/mg protein (CAT); nmol NADPH oxidized/min/mg protein (GPx); nmol/mg protein (DCF); nmol SH/mg
protein (total thiol content). There were no significant differences in the activity of antioxidant enzymes in males: Non-handled:
SOD 14.45+-2.07; CAT 1.24+-0.30; GPx 36.59+-6.11; DCF 6.26+-0.32, and total thiol content 57.14+-2.35. Handled: SOD
14.45+-2.07; CAT 1.21+-0.25; GPx 39.52+-7.50; DCF 6.60+-0.61, and total thiol content 61.10+-1.18. For females, there were
no differences either: Non-handled: SOD 14.88+-2.14; CAT 2.30+-0.33; GPx 47.03+-1.79; the DCF 3.95+-0.30, and total thiol
content 42.57+-3.89. Handled: SOD 16.96+-2.20; CAT 2.34+-0.14 ; GPx 46.82+-1.20; the DCF 4.44+-0.33, and total thiol
content 45.81+-3.39. In all cases p>0.05.
Conclusions:
These results suggest that there is not influence of neonatal handling in oxidative stress parameters measured in the striatum of
adult male and female rats. However, this process may be occurring in other brain structures. For example, it should be observed
that previous studies have shown increased DNA breaks index in hippocampus of male animals that were handled in the neonatal
period. Therefore, this opens up prospects for further evaluation of these animals.
Keywords: Oxidative stress, neonatal handling, striatum
Financial Support: CNPq, PRONEX-FAPERGS
Resumo:23-166
HISTOPATHOLOGIC ANALYSIS IN STRIATUM AND HIPPOCAMPUS OF MICE TREATED ACUTELY WITH
SESQUITERPENE FARNESOL
Pereira, W. B. 1; Batista, R. B. 1; Torres, P. D. A. 1; Junior, W. M. D. O. 1; Paulo, L. L. 1; Morais, L. C. S. L.

D. 1; Sousa, D. P. D. 2; Freitas, R. M. 3; Almeida, R. N. 1
1
Lab. de Tecnologia Farmacutica/Univers. Federal da Paraba, UFPB
2
Departamento de Fisiologia/ Univers. Federal de Sergipe, UFS
3
Departamento de Farmacologia/ Universidade Federal do Piau, UFPI
Objectives:
Essential oils are products with complex composition extracted from aromatic plants that have various pharmaceutical
applications such as antiseptic, antimicrobial, sedative, anti-inflammatory and spasmolitic properties. The sesquiterpene farnesol
is a natural compound found in aromatic plants as lemon grass and chamomile (Plant Cell, 7; 1015-1026, 1995). The aim of this
study was to evaluate histopathological alteration in the brain of mice acutely treated with Farnesol.
Male Swiss mice (60 days old and weighing 25-30g) were divided into 4 groups (N = 6) that were treated intraperitoneally (ip)
with tween 80 and saline 0.9% (control) or farnesol 50, 100 and 200 mg/ kg (experimental groups). After 24h of the treatments,
all the animals were anesthetized with sodium pentobarbital (30 mg/ kg, ip), the brains were removed and fixed in 10% formalin
for 72 h to the histopathological analysis. Sagittal sections were obtained from the mammillary bodies. For the study by optical
microscopy, sections of 10 were stained with hematoxylin-eosin (HE). Brain injury was defined as at least 50% of
histopathological alteration in each area examined (hippocampus and striatum). The groups treated with farnesol, only showed an
impairment in hippocampal at dose of 200 mg/kg, corresponding to 12%. Regarding the histopathological analysis of the
striatum, there was no evidence of any type of injury or impairment resulting from the acute treatment with all farnesol doses.
Conclusions:
Our results indicate that acute treatment with farnesol does not induced significant changes in the hippocampus and striatum of
mice.
Keywords: Farnesol, Hippocampus, Histopathologic, Striatum
Financial Support: CNPq/ UFPB
Resumo:23-167
EFFECTS OF DIAZEPAM AND BIPERIDEN ON MEMORY AND ANXIETY IN MICE ASSESSED IN THE
ELEVATED T-MAZE TASK
Asth, L. ; Andr, E. ; Soares, V. P. ; Gavioli, E. C.

Universidade Federal do Rio Grande do Norte, UFRN
Objectives:
The elevated T-maze (ETM) is an apparatus derived from the elevated plus-maze test, which is widely used to evaluate anxiety in
rodents (Neurosci Biobehav Rev. 29(8):1193-205, 2005). Considering that anxiety is a biasing factor in most of the available
models of memory, this study was aimed to investigate whether the ETM is able to assess memory and anxiety simultaneously in
mice. The ETM consists of one enclosed and two open arms. The procedure is based on an experimental protocol previously
proposed for rats (Pharmacol Biochem Behav.63(1):63-9, 1999), in which mice were exposed to the enclosed arm as many times
as needed to stay 300 s. One day later (test session), memory is assessed by reexposing the mouse to the enclosed arm, and the
latency to enter the open arm with the four paws was recorded. The assessment of learning and memory processes using the ETM
task is based on the unconditioned avoidance to open spaces learned during the training session, in which animals possibly learn
to discriminate, and then avoid, the unprotected areas of the apparatus.
Male Swiss mice weighting 2530 g were housed in groups of fifteen to twenty per cage (33x40x17 cm) with food and water ad
libitum. Animals were maintained under constant temperature (232C), and under a 12-h light/dark cycle (lights on at 06:00 h).
The anxiolytic diazepam (DZP; 1 and 2 mg/kg, ip), and the amnestic biperiden (BPR; 1 mg/kg, sc) were used by injecting at two
distinct times: pretraining and pretest. Pretraining administration of BPR (n=10) and DZP (DZP1, n=12 and DZP2, n=7) trended
to increase the number of trials to reach the avoidance criterion (C= 6.90.9, BPR= 9.91.2, p=0.06; C=5.71.1, DZP1=8.81.0,
DZP2=10.01.8, p=0.09), suggesting learning impairment. Pretraining injection of BPR did not affect the spontaneous increase in
the latency to enter the open arm between trials (p>0.05), while DZP, in the trial 1 and 2, reduced this parameter compared to
control (trial 1: C=26.36.5, DZP1=9.31.6*, DZP2=15.23.3, *p=0.02; trial 2: C=59.917.6, DZP1=17.13.1*,
DZP2=12.24.7*, *p0.05), suggesting anxiolysis. In the test session, pretraining injection of BPR and DZP 2, but not DZP1,
reduced latencies to enter the open arm (C= 197.734.6, BPR= 42.928.7*, *p0.05 for both).
Conclusions:
The ETM is an animal model able to assess and discriminate simultaneously the anxiolytic and amnestic effects of drugs in mice.
Further experiments are required to investigate in the ETM the effects of memory enhancer and anxiogenic drugs.
Keywords: memory, anxiety, animal model, mouse
Financial Support: CNPq (Ed. Universal 476832/2009-8; Bolsa Mestrado: 133814/2010-6)
Resumo:23-168
CENTRAL AND PERIPHERAL ADMINISTRATION OF LPS ALTERS DIFFERENTLY THE S100B LEVELS IN
SERUM AND CEREBROSPINAL FLUID
Negri, E. ; Tortorelli, L. S. ; da R, C. F. ; Galland, F. ; Engelke, D. S. ; Guerra, M. C. ; Leite, M. C. ;

Rodrigues, L. ; Gonalves, C. A.
Objectives:
S100B is a calcium binding protein mainly expressed and secreted by astrocytes in the central nervous system and has been
described as a marker of astroglial activation, measured in cerebrospinal fluid (CSF) and serum. However, adipocytes contribute
to the serum levels of this protein. Thus, it is important to know the factors that influence the apparent compartmentalization of
this protein between serum and cerebrospinal fluid (CSF) to understanding the significance of changes. Our objective was to
evaluate serum and CSF levels of S100B in response to acute stimulation with lipopolysaccharide (LPS) injected centrally or
peripherally.
Anesthetized adult wistar rats received 10 L ICV of 2.5 ug/uL LPS or phosphate-buffered saline (control). CSF,collected by
cisterna magna puncture, as well as blood, collected by intracardiac puncture, at 30 min or 24 h after. S100B and TNF alfa
content was measured by ELISA. A significant increase in CSF S100B was observed in 30 min (205%, p=0.009, n=5) and in 24 h
(301%, p=0.003, n=5) after ICV LPS administration, without significant changes in S100B serum content. Interestingly, when
rats received IP LPS (250 g/Kg body) they also exhibited an increase in CSF S100B in 30 min (250%, p=0.04, n=5), but not in
24 h, and again no significant changes in serum S100B were observed when compared with controls that received phosphatebuffered saline. We measured a classical inflammatory cytokine, TNF, in response to LPS in vivo to confirm the activity of this
compound and compare to that obtained with S100B protein. Differently from S100B, 30 min and 24 h after IP administration of
LPS (approximately 75 ug), we observed an increase in serum TNF 30 min (192.1 pg/mL 97.2) in relation to control (3.4
pg/mL 1) , and in serum TNF alfa 24 h ( 2.6 pg/mL 0.2) in relation to control (1.1 pg/mL 0.4) but not in CSF. When LPS
(25 ug) was administered ICV we found an early and transient increase in serum at 30 min (121.6 pg/mL 40) in relation to
control (0.7 pg/mL 0.3) and a later increase in CSF at 24 h (19.6 pg/mL 4.5) in relation to control ( 1.7 pg/mL 1.2).
Conclusions:
Our results suggest a brain specific LPS-induced release of S100B, i.e., peripheral immune cells stimulated by LPS did not
release or cause a detectable S100B release from potential extra-cerebral sources of S100B (e.g. adipocytes).Together, these data
contribute to our understanding of the effect of LPS on astrocytes, particularly on S100B secretion and help us to interpret
cerebrospinal fluid and serum changes of this protein in brain inflammatory disorders.
Keywords: Astrocytes, Lipopolysaccharide, Neuroinflammation, S100B
Financial Support: CNPq, CAPES, FAPERGS e rede IBN-NET
Resumo:23-169
INTRACEREBROVENTRICULAR GMP ADMINISTRATION INDUCES ANXIOLYTIC-LIKE EFFECT IN RATS.
Almeida, R. F. ; Comassetto, D. D. ; Ramos, D. B. ; Junior, V. H. C. ; Ganzella, M. ; Souza, D. O.

Objectives:
In a recent study, we demonstrated that a systemic administration of GMP induces anxiolytic-like effect similar to diazepam (Dz)
in the light/dark and elevated plus-maze (EPM) tasks in rats (Pharmacol Biochem Behav. 96(3):306-11, 2010). In the present
work, we evaluated the effects of a central GMP administration and the dependence of its conversion to guanosine (GUO) for the
anxiolytic-like effect.

Data were analyzed by one-way ANOVA followed by pos hoc Tukey test p
Conclusions:
The present work shows that both central and systemic GMP administration induces anxiolytic-like effect. Additionally, our
results suggest that GMP effect may be dependent of its conversion to GUO since CSF GUO concentration was increased after
systemic GMP administration. However more studies are necessary to elucidate the exactly mechanism of GMP anxiolytic-like
effect.
Keywords: Adenosine, Anxiety, Diazepam, GMP, Guanosine
Financial Support: CNPq, Capes, INCT, IBN.Net.
Resumo:23-170
EVALUATION OF A LIPOSOME DIHYDROPYRIDINES (LCN) DERIVATIVE ON FORCED SWIMMING TEST
Moreno, L. C. G. A. I. 1; Costa, J. P. 1; Costa, D. A. 1; Sabino, E. B. 1; Magalhes, N. S. S. 2; Freitas, R. M.

1
; Santos, ,. H. M. L. R. 1
1
Universidade Federal do Piau, UFPI
2
Objectives:
Increasing concentrations of intracellular Ca++ cause contraction of vascular muscle cells and the heart. In this context, some
drugs act by binding these channels in order to promote a competitive inhibition. Thus, the channel blockers of Ca++ act to
reduce the excitability rate of the heart and promote vasodilation. An example of blockers is dihydropyridines group. These
molecules have a high lipophilicity and readily cross the blood brain barrier, remaining in the brain. It is known that a more
effective cerebral circulation leads to improvements in many diseases. However, in order to produce beneficial effects, these
drugs should be administered at high doses, causing several side effects. These disconforts can be minimized by using liposomes,
since they promote a controlled release of the drug, increasing its bioavailability and reducing its toxicity. The objective the
present study was to develop a liposomal formulation containing a vasodilator belonging to dihydropyridines class as well as to
evaluated its pharmacological potential on forced swimming test.
The preparation, known as LCN, was produced by using soybean phosphatidylcholine lipids and cholesterol by the method of
forming lipid film followed by sonication. The characterization was performed using specific techniques of quality control for
determination of physicochemical parameters. LCN showed particle size of 121.93 0.60 nm, polydispersity 0.297 0.02, zeta
potential -5.32 1.29, pH 7.3, and yield rate drug content of 100%. After their characterization, the formulation was administered
in three groups of seven adult male Swiss mice at doses of 0.1 (LCN 0.1), 1.0 (LCN 1) and 10 (LCN 10) mg/kg. A fourth group
was given 0.9% saline solution (control group) and a fifth received imipramine at a dose of 50 mg / kg (positive control). After 30
min from administration, forced swimming test was performed and immobility time was determined in seconds. Mean immobility
time to LCN 0.1, LCN 1 and LCN 10 were 9.14 5.68, 7.57 3.66 and 1.0 0.72, respectively. The results indicate a possible
antidepressant effect of the formulation, since the immobility time of the doses were significantly lower than the average of both
control (224.3 0.86) and imipramine-treated animals (75.00 0.68), suggesting a possible blocking effect in the reuptake of
norepinephrine and serotonin.
Conclusions:
By the results of the study, we cannot conclude that LCN have antidepressant effect, but we can suggest an action on central
nervous system and further investigations on LCN with associations of antidepressants already described in literature should be
carried out.
Keywords: ANTIDEPRESSANT EFFECT, DIHYDROPYRIDINES, FORCED SWIMMING TEST , LIPOSOME
Resumo:23-171
EVALUATION OF THE EFFECTS OF CANNABIDIOL IN A KETAMINE-INDUCED SCHIZOPHRENIA MODEL IN
RATS
de Ross, J. B. 1; Hallak, J. E. C. 1; Crippa, J. A. S. 1; Zuardi, A. W. 1; Leite, J. P. 1; Romcy-pereira, R. N. 2,3

1
Neurocincias e Cincias do Comportamento/FMRP, USP
2
Instituto do Crebro, UFRN
3
Instituto Internacional de Neurocincias de Natal, IINN-ELS
Objectives:
Schizophrenia is a neuropathological disorder with a poorly understood etiology, manifested by positive, negative and cognitive
symptoms. Blockade of glutamatergic NMDA receptors can generate psychotic symptoms in healthy subjects and exacerbate
them in schizophrenic patients. In rodents, they produce sensori-motor deficits, hyperlocomotion and alterations in cortical
gamma oscillations. Recent studies have shown that Cannabis sativa components have a substantial therapeutic potential,
suggesting for cannabidiol (CBD) an atypical antipsychotic profile, similar to the standard drug clozapine (CZP). Therefore, the
objective of this study was to investigate the behavioral and physiological effects of cannabidiol administration in a rat model of
acute psychosis induced by NMDA receptors hypofunction through S(+)ketamine injection.
Adult male Wistar rats were first tested for prepulse inhibition of the startle reflex (PPI) and only those who had PPI greater
than 50% were used for the experiments. Each group (n=10) received an intraperitoneal injection of vehicle (1 mlkg; corn oil),
CZP 2.5 mgkg (CZP2.5) or CBD 30 mgkg (CBD30). After 30 minutes, they were placed in an open field and exploratory
behaviors were recorded for 40 minutes. On the 20th minute, rats received an injection of saline or S(+)ketamine 30 mgkg
(KET30). The recording was divided into 20 blocks of 2 minutes for posterior analysis. Four parameters were considered:
traveled distance, average speed, time spent in the center of the open field and rearing frequency. At the end of this experiment,
animals were tested using a PPI protocol for analysis of the startle reflex amplitude (pulses of 120 dB) and the %PPI evaluated at
3 prepulse distinct intensities (68, 71 e 77 dB). For PPI experiment, oneway ANOVA followed by StudentNewmanKeuls
test revealed that CBD30 group had a %PPI increase on 71 dB pre-pulse intensity, compared to vehicle+KET30 group (p =
0.018), but had no significant difference in relation to CZP2.5+KET30 group (p = 0.877). In the exploratory behavior experiment,
twoway RM ANOVA followed by StudentNewmanKeuls revealed a reduction in distance traveled and in average speed in
the CBD30+KET30 group, on the 22th, 24th, 30th, 36th and 40th minutes compared to vehicle+KET30 group (p
Conclusions:
Cannabidiol was able to attenuate psychotomimetic effects induced by S(+)ketamine in rats, considering the reversal of PPI
deficit, similar to clozapine, and decreased hyperlocomotion, observed by reduction in traveled distance and average speed.
Besides, the increase in time spent in the center and in rearing frequency indicates an anxiolytic effect. The present study may
contribute to a better understanding of the neurochemical mechanisms implicated in the ketamine model of schizophrenia.
Keywords: schizophrenia, cannabidiol, ketamine model, pre-pulse inhibition, exploratory behavior
Financial Support: Capes, CNPq, Fapesp and FAEPA
Resumo:23-172
STRESS CHANGES ON NEUTROPHIL ACTIVITY RELY ON GENDER AND HIERARCHY IN DOGS
Akamine, A. T. 1; Pinheiro, M. L. 1; Ribeiro, A. 1; Ferraz-de-paula, V. 1; Almeida, V. I. 1; Taricano, I. D. 2;

Costa-pinto, F. A. 1
1
Universidade de So Paulo, VPT/FMVZ/USP
2
Inst. de Educao, Pesquisa e Desen. Tecnologico Royal, Instituto Royal
Objectives:
Hierarchy influences vulnerability to disease. Reorganization of hierarchical levels in a group of dogs can be differentially
stressful to each individual. Effects of stress on immunity have been widely assessed in rodents but not in dogs, which are the
experimental subject of choice in toxicological and preclinical studies. Thus, this study aims to identify the effects of stress on the
immune system of dogs with vary in gender and hierarchy.
We employed transportation (short-term) or hierarchical reestablishment (long-term) as putative stressors in dogs and assessed
their effects on circulating neutrophils from thirty 4-month-old beagle dogs (fifteen males and fifteen females). Blood samples
collected before (t0), after 1h (t1), and 15d (t2) of relocation for the assessment of WBC and neutrophil activity. Transportation
increased the percentage of circulating neutrophils (p<.05).
Conclusions:
The paradigm employed here influenced differentially males compared to female dogs regarding neutrophil counts and activity.
This could underlie differences in vulnerability commonly observed in dogs. Further parameters such as cortisol levels and the
ratio CD4/CD8 T cells are under investigation.
Keywords: neuroimmunomodulation, neutrophils, stress, dog, hierarchy
Financial Support: FAPESP - 09/51886-3, CNPQ e CAPES.
Resumo:23-173
ACCELERATING MEMORY SYSTEM CONSOLIDATION BY NEW LEARNING EXPERIENCE
Cassini, L. ; Haubrich, J. ; de Oliveira Alvares, L. ; Santana, F. ; Crestani, A. P. ; Sierra, R. O; Diehl, F. ;

Quillfeldt, J. A.
Departamento de Biofsica, UFRGS
Objectives:
After initial encoding, memory undergoes a time-dependent reorganization process, by which its retrieval becomes independent
of Hippocampus (HPC) and sustained by cortical regions, such as the Anterior Cingulate Cortex (ACC). This phenomenon is
known as system consolidation and its mechanism still eludes us. Here we studied the influence of new learning experiences on
system consolidation of a contextual fear conditioning (CFC) memory.
Male Wistar rats were trained in CFC task and tested 1-45 days later, depending on the experiment. To examine whether the
memory retrieval was dependent or not of a target structure (HPC or ACC), animals received microinjections of a classical
amnesic drug (Muscimol 1ug/ul) 15min before the test session. Student t-Test was used for comparison between two groups and
One-way ANOVA for more than two. First we tested how much time the CFC memory takes to become HPC-independent and
ACC-dependent, and we found that this process takes about 40 days. Next we evaluated whether the acquisition of new memories
during this period (Object Recognition and Water Maze) could influence the rate of CFC system consolidation. We found that
when animals were submitted to new learning after training, the CFC memory took only 20 days to become HPC-independent
and ACC-dependent. As system consolidation was much faster at these conditions, we tested if the precision of CFC memory was
maintained or not. For that, animals were tested at a very similar context of the training, and they distinguished it from the
original one as good as the control group. Finally we tested if this phenomenon was specific to the CFC task, or if it is a more
general mechanism of memory processing. The same protocol was performed, but instead of CFC, the Inhibitory Avoidance (IA)
task was used. Again, when animals acquired new memories after training, the IA memory become hippocampus-independent
earlier.
Conclusions:
The newly acquired information probably accelerated the CFC system consolidation, leading to its reorganization and storage in
extra-hippocampal regions earlier. Moreover, this faster consolidation did not affect the precision of memory and was not specific
to one kind of memory. Our findings suggest that System Consolidation is a dynamic, non-stereotyped process that may be
accelerated by interposing intense cognitive activity between the training and the test session.
Keywords: Memory, System Consolidation, Hippocampus, Anterior Cingulate Cortex, Fear Conditioning
Financial Support: CAPES, CNPq, FINEP, FAPERGS, IFS.
Resumo:23-174
EVALUATION OF PERFORMANCE PARAMETERS, IMMUNITY AND BACTERIA MIGRATION IN HEATSTRESSED BROILER CHICKENS INFECTED BY SALMONELLA ENTERITIDIS
Quinteiro-filho, W. M. ; Gomes, A. S. ; Ribeiro, A. ; Ferraz-de-paula, V. ; Pinheiro, M. L. ; Astolfi-ferreira,

C. S. ; Ferreira, A. J. P. ; Palermo-neto, J.
Departamento de Patologia / FMVZ, USP
Objectives:
Stressful stimuli are known to change the immune function. A relevant environmental stress to the poultry production is the heat
stress. The aim of our work was search for effects of heat stress on performance parameters, corticosterone serum level, IgA
plasma level, IFN-lambda levels and bacteria migration to spleen of broilers chickens infected with Salmonella enteritidis,
searching for a neuroimmune relationship.
Methods: One hundred sixty chickens were housed in 2 rooms, infected or not with Salmonella enteritidis (106UFC/animal).
Animals were divided in four groups: control (C); heat stress (HS); salmonella+control (SC); and salmonella+heat stress (SHS).
Controls group (C and SC) were kept under 211 C of environmental temperature. Both HS and SHS groups were exposed to
311C (10h/day) from day 35 to 42. The production parameters were determinate by weight gain (WG), feed consumption
(FCo) and feed conversion (FC) during heat stress period. At 42nd day, 10 birds per group were randomly selected to compose all
groups. Blood samples were harvested to analyze corticosterone serum levels, IgA and IFN-lambda plasma levels by Enzyme
Linked Immunosorbent Assay (ELISA) and spleen were harvested to analyze bacteria migration by microbiology assay. Results:
We observed that heat stress per se decreased performance parameters such as body weight (g) (C=622.739.23;
HS=442.544.81*; SC=618.523.85; SHS=374.749.22*, pSalmonella FC (C=2.450.14; HS=2.510.20; SC=2.160.04;
SHS=2.730.23*, p
Conclusions:
Thus, we believe that heat stress impaired performance parameters, moreover the heat stress and the Salmonella infection
together impair more intensely these parameters, principally feed conversion. Furthermore, heat stress induced CNS activation
increasing corticosterone levels, which in turn decreased the immune system activity and lead to impairment of mucosal barrier,
which in turn increased chickens susceptibility to Salmonella migration to the spleen.
Keywords: heat stress, Salmonella, Immunity, corticosterone, HPA-axis
Financial Support: Fapesp, CNPq
Resumo:23-175
IMIPRAMINE TREATMENT IN MALE MICE PREVENTS HIERARCHY DESTABILIZATION INDUCED BY
LIPOPOLYSSACHARIDE ADMINISTRATION
Cohn, D. W. H. ; Kinishita, D. ; Palermo-neto, J.

Depto de Patplogia FMVZ, USP
Objectives:
Lipopolyssacharide (LPS) administration in the dominant mice promotes destabilization of social hierarchy within a dyad. Recent
works show a relationship between sickness behavior induced by LPS and depression. In this work we sought to determine if
antidepressants could prevent social consequences caused by sickness behavior display by the dominant mice.
Social hierarchy in dyads of male mice was evaluated during 5 weeks. Treatments were applied on week 2, when mice were
placed on observation cages and had their behavior scored by ethological methods. There were four experimental groups: 1)
Saline: dominant mouse was treated with saline for 3 consecutive days; 2) Imipramine + Saline: dominant mouse was treated
with imipramine (20 mg/kg) for 3 days; 3) Saline + LPS: dominant was treated with LPS (400 g/kg); 4) Imipramine + LPS:
dominant was treated with imipramine and LPS (same doses as above) for 3 consecutive days. LPS treatment was responsible for
destabilization of social dominance. Such destabilization was prevented by antidepressant treatment. In spite of the fact that LPS
treated mice still showed signs of sickness, imipramine was responsible for greater motivation to engage in social behavior which
might explain obtained data.
Conclusions:
Imipramine prevents relevant social consequences of sickness behavior display. This results could provide a new paradigm to the
study of the behavioral neurobiology of depression.
Keywords: LPS, sickness behavior, depression, social behavior , aggression
Financial Support: FAPESP (proc. 09/52419 e 2009/ 518866-3)
Resumo:23-176
FOLIC ACID PREVENTS THE DEPRESSIVE-LIKE BEHAVIOR ELICITED BY THE PRO-INFLAMMATORY
CYTOKINE TNF-ALPHA IN THE MOUSE TAIL SUSPENSION TEST
Budni, J. ; Moretti, M. ; Freitas, A. E. ; Neis, V. B. ; Ribeiro, C. M. ; Balen, G. O. ; Amorim, M. B. ;

Rodrigues, A. L. S.
Bioqumica/ Universidade Federal de Santa Catarina, UFSC
Objectives:
Several studies have indicated that folic acid plays a role in the pathophysiology of depression, a disorder that may involve an
activation of the inflammatory response system. This study investigated the ability of folic acid alone or combined with
antidepressants, MK-801 and 7-nitroindazole to reverse the depressive-like behavior induced by the pro-inflammatory cytokine
tumor necrosis factor-alpha (TNF-alpha)
Adult (60 days old, 30-40 g) female Swiss mice (n=8 per group) were used in this study. Immobility time and locomotor activity
were evaluated in the tail suspension test (TST) and in an open-field test, respectively, in a 6-min session. The data were analysed
by two-way ANOVA followed by Newman-Keuls test when appropriate. Folic acid administered orally at 10 and 50 mg/kg (but
not at 1 mg/kg) caused a reduction of the immobility time (28.222.07% and 23.281.31%, respectively, when compared with
control 100%) in the TST. A sub-effective dose of folic acid (1 mg/kg) combined with sub-effective doses of fluoxetine (1 mg/kg,
p.o., serotonin reuptake inhibitor), bupropion (1 mg/kg, p.o., dopamine reuptake inhibitor), imipramine (0.1 mg/kg, p.o., tricyclic
antidepressant), MK-801 (0.001 mg/kg, p.o, non-competitive NMDA receptor antagonist) or 7-nitroindazole (25 mg/kg, i.p.,
neuronal nitric oxide synthase inhibitor) reduced the immobility time in the TST (49.564.58%, 27.301.87%, 31.764.48%,
38.585.22% and 36.402.41%, respectively X control group 100%), suggesting a synergistic antidepressant-like effect. The
administration of TNF-alpha (0.001 fg/site, i.c.v.) increased the immobility time in the TST, indicating a depressive-like effect
(37.110,89% X control group 100%) that was reversed by folic acid (1, 10 and 50 mg/kg, p.o.), fluoxetine, bupropion,
imipramine, MK-801 and 7-nitroindazole (100.634.68%, 78.165.78%, 84.864.82%, 97.362.88%, 93.384.52%,
98.696.32%, 96.996.19%, 89.994.52% respectively X control group 100%). Additionally, folic acid in combination with
fluoxetine, bupropion, MK-801 or 7-nitroindazole produced a synergistic antidepressant-like effect (77.797.19%, 76.014.01%,
64.138.18% and 76.057.00%, respectively X control group 100%) against the increase of immobility time elicited by TNFalpha. Neither drug alone or in combination with folic acid altered the locomotor activity.
Conclusions:
These results indicate that TNF-alpha induces a depressant-like effect in the mouse TST that was prevented by folic acid alone or
in combination with antidepressants, MK-801 and 7-nitroindazole. Results suggest that the reported effects of folic acid may
involve inhibition of NMDA receptors and nitric oxide synthase and that folic acid associated with antidepressants might be
useful in treating inflammation-associated depression
Keywords: Folic acid, TNF-Alpha, Tail suspension test, Depression, Antidepressant
Financial Support: CAPES, CNPq and FINEP-IBN-Net
Resumo:23-177
BEHAVIORAL AND METABOLIC EFFECTS OF CHRONIC MEMANTINE ADMINISTRATION IN ADULT
MALES MICE
Torrez, V. R. ; Zimmer, E. R. ; Kalinine, E. ; Zenki, K. C. ; Souza, D. O. ; Portela, L. V.

Bioqumica/ Universidade Federal do Rio Grande do Sul, UFRGS
Objectives:
Memantine (MN) is an uncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist, which is clinically used in the
treatment of moderate to severe cases of Alzheimers disease (AD). The NMDA receptor family is a group of excitatory
ionotropic receptors involved in the glutamatergic neurotransmission. Glutamate is the major excitatory neurotransmitter in
mammalian brain and plays important roles in the central nervous system (CNS). Glutamate mediates various brain responses,
such as anxiety, fear and appetite, by regulating neuronal excitability in several brain structures. Nevertheless, under pathological
conditions, increased glutamate levels in the synaptic cleft leads to damage by excitotoxicity. Several studies have showed that
MN is neuroprotective against neural damage; however, behavioral and metabolic effects still need additional investigations. The
main goal of this study was to investigate the effects of chronic oral administration of MN on anxiety-like behavior and metabolic
parameters in mice.
Male CF1 mice 2 months old (n=64) were divided in four groups: Control (CO), MN5, MN10 and MN20, which received saline,
5, 10 and 20 mg/kg of memantine, respectively. They were treated by gavage once daily for 23 days. The animals were submitted
to behavioral tasks: open field, light-dark box and elevated plus maze. Memantine group showed a tendency to increase
spontaneous locomotion and exploratory activity (n=16 per group, p=0.0540). In the elevated plus maze, memantine caused an
anxiogenic behavior at lower doses (n=10 per group, **p=0.0047), and, in the light-dark box, all MN groups presented
anxiogenic behavior (n=16 per group, **p=0.0013). We also analyzed the food intake, body weight and visceral fat pads. There
were no differences in food intake (n=16 per group, p=0.0862), body weight (n=16 per group, p =0.9982) and visceral fat pads
(n=16 per group, p=0.0791). Animals were sacrificed by decapitation and hippocampus and cortex were quickly dissected out on
ice. Brain slices were used for glutamate uptake and homogenates for western blotting. In the cortex, MN10 and 20 decreased
glutamate uptake (n=10 per group, **p=0.0012), while in the hippocampus no changes were found (n=10 per group, p= 0.6744).
All the data were analyzed by one-way ANOVA, followed by Bonferronis post-hoc test, except for food intake and body weight,
which were analyzed by two-way ANOVA, followed by Bonferronis post-hoc test. Differences between groups were considered
statistically significant if P < 0.05.
Conclusions:
Memantine is already in use by humans and shows safety and security, and, in our study, did not alter metabolic parameters.
Besides, our findings suggest that chronic administration of memantine causes anxiogenic behavior associated to decrease of
cortical glutamate uptake.
Keywords: anxiety, body weight, glutamate uptake, memantine
Financial Support: CNPq, Capes, INCTEN e Fapergs
Resumo:23-178
EVALUATION OF ANXIOLYTIC ACTIVITY OF ETHANOLIC EXTRACT FROM TERMINALIA FAGIFOLIA IN
MICE
Pereira, C. F. C. 1; Oliveira, J. M. G. 2; Nascimento, L. F. M. 1; S, M. C. 2; Sales, P. A. B. 1; Nunes, P. H.

M. 2; Lira, S. R. S. 1; Fernandes, R. M. 1,2; Costa, A. P. R. 1,2; Neto, J. D. N. 2
1
Departamento de Morfofisiologia Veterinria, UFPI
2
Ncleo de Pesquisas em Plantas Medicinais, UFPI
Objectives:
One of most common mental disorders in humans is the pathological anxiety, which may be minimized by anxiolytic substances,
such as benzodiazepine drugs. The main problem with these drugs is its dose-dependency effect and adverses reactions. In search
of new compounds with anxiolytic proprieties, many plant and its products have been studied aimed a solution to the wide use of
drugs in the treatment of anxiety. Terminalia fagifolia is a tree of the savanna, known popularly as capito, capito-do-mato,
mirindiba, chapadeiro, cachaporra-do-gentio and pau-de-bicho. Fractionation of ethanolic extract from T. fagifolia led to the
isolation and structural identification of a three triterpenes, which are compounds with known sedative effects on the Central
Nervous System. Therefore, the aim of this paper was evaluate the anxiolytic potential of ethanolic extract of stem bark from
Terminalia fagifolia in mice.
The plant material was dried at room temperature, pulverized in a grinder of knives and placed in dark plastic bottles. The
ethanolic extract from T. fagifolia (ETOHTf) was prepared by maceration with absolute ethanol by successive extractions in a 72hour interval between each extraction, for four consecutive times, concentrated on rotary evaporator at 50C and then lyophilized.
Were used 30 albino mice, adult, male, weighing 25 to 35 grams and kept under controlled conditions, receiving water and food
ad libitum. Were used groups of six mice pre-treated orally with distilled water (C), with ETOHTf (250, 500 or 750 mg/kg) or
diazepam (2 mg/kg intraperitoneal). The animals were placed 30 minutes after treatment in an open-field and were observed for 5
minutes, and was evaluated the exploratory activity of animals (total number of crossings), the total number of rearings, number
of self-cleaning, number of dung pats and the time that the animals remained immobile. The results were analyzed comparing the
differences between means by one-way ANOVA test followed by Tukey post-test (p
Conclusions:
The ethanolic extract from stem bark of Terminalia fagifolia did not show anxiolytic activity when compared statistically to
control, despite having reduced the number of rearings and increased the time that animals remained immobile.
Keywords: anxiolytic, open-field , Terminalia
Financial Support: Universidade Federal do Piau
Resumo:23-179
HIGH DOSE OF MODAFINIL INDUCES CONDITIONED PLACE PREFERENCE IN MICE
Boroi, A. R. 1; Wuo-silva, R. 1; Hollais, A. W. 1; Santos, R. 1; Ribeiro, L. T. C. 1; Saito, L. P. 1; Fukushiro,

D. F. 1; Bittencourt, L. R. 2; Tufik, S. 2; Frussa-filho, R. 1
1
Department of Pharmacology, UNIFESP
2
Department of Psychobiology, UNIFESP
Objectives:
Modafinil (MOD) is a wake-promoting agent used in the treatment of excessive daytime sleepiness in narcolepsy. However,
whereas some papers suggest an addictive effect of MOD, other studies have shown that a high (300 mg/kg) dose of this drug
blocks morphine seeking behaviors. The purpose of the present work was to investigate a potential addictive effect of this high
dose of MOD on the conditioned place preference paradigm.
Twenty-two Swiss male were randomly allocated into 2 groups (n=11): vehicle (VEH) and MOD. Conditioning sessions were
conducted twice daily with a minimum interval of 6 hours from one to another. Animals of the MOD were injected with vehicle
or MOD (i.p.) and 30 min later confined to one of the compartments of the place preference apparatus for 10 minutes. Treatment
compartment was counterbalanced for vehicle and MOD, as well as the presentation order of vehicle and MOD. Animals of the
VEH group received vehicle in both compartments. For test sessions (one day after last conditioning session), uninjected mice
were allowed free access to both compartments of the place preference apparatus for 15 minutes. The paired T-test showed that
mice of the MOD group presented increased time spent in the drug-paired compartment when compared with vehicle-paired
compartment [t(11)= 4.443, P0.05]. The unpaired T-test showed that CPP score was significantly higher for the MOD group than
for the VEH group [t(20) = 2.6, P
Conclusions:
Our results showed that a high dose of modafinil induced a conditioned place preference in mice, indicating the potential
addictive effect of this drug.
Keywords: Addiction, Modafinil, Place Preference
Financial Support: FAPESP, CNPq, CAPES e FIPE.
Resumo:23-180
RELATIONSHIP BETWEEN MEMORY AND ALPHA7 NICOTINIC RECEPTOR DENSITY IN AN ANIMAL
MODEL OF SEVERE ALZHEIMERS DISEASE.
Schwe, N. M. 1; Monteiro-silva, K. C. 2; Oliveira, E. M. 3; Caetano, A. L. 2; Buck, H. S. 2; Viel, T. A. 1,3

1
Institute of Biomedical Sciences, USP
2
Department of Physiological Sciences, FCMSCSP
3
School of Arts, Sciences and Humanities, USP
Objectives:
Alzheimers disease (AD) is characterized by amyloid-beta (A) plaques in brain and leads to cognitive impairment. Initially,
memory process is affected leading to a progressive neurodegeneration along the course of the disease. Previous studies of our
research group showed that chronic infusion with A peptide, per se, caused an increase in 7 nAChR density in some
hippocampal areas. This receptor modulates long term potentiation and, therefore, memory processes. In the first part of the
present work, transgenic mice showed similar performance in spatial memory and decreased performance in aversive-related
memory when compared to wild-type mice. The aim of this study was to evaluate 7 nAChR density in these animals
hippocampus.
3-month-old male C57Bl6 (WT, n=10) and Tg(PDGFB-APPSwInd) (TG, n=8) mice were kept in standard cages with water and
food ad libitum until they complete 7 months of age. Behavioral tests were performed when the mice were 5 and 7 months old. In
evaluation of spatial memory at Barnes maze (5 blocks of two trials), it was observed that TG and WT mice learned the task and
had the same performance with no statistical difference between them at 5 or 7 months of age. At inhibitory avoidance test (0,5
mA, 2 sec, max latency of 300 sec) 5-month-old WT and TG mice showed difference in latency in test session (TS) performed
24h after the acquisition session(AS), indicating that learning occurred (WT:AS: 12.2 [9.0/20.3] sec and TS: 300.0 [298.0/300.0]
sec, p < 0.01; TG: AS: 30.6 [11.5/39.1 sec and TS: 300.0 [180.7/300.0] sec, p < 0.05). At 7 months old, WT mice maintained
their performance (298.0 [140.9/299.0]), while TG showed a significant reduced latency (29.6 [15.6/51.4], p < 0.001), indicating
that they did not remember the task.. At the end, brains were extracted and 20 m slices were obtained on a cryostat (18C).
Autoradiography (ARG) for 7 nAChR was conducted using [125I]--bungarotoxin (5 nM, 90 min, 25C). Non-specific binding
was assessed using 2 M of the unlabelled toxin and corresponded to 5% of total binding. TG presented greater 7 nAChR
density than WT in the following hippocampal areas: granular layer of dentate gyrus (WT: 7.4 0.1 fmol/mg; TG: 8.3 0.2
fmol/mg; p < 0.05), pyramidal cells (WT: 9.9 0.32 fmol/mg; TG: 11.8 0.3 fmol/mg, p < 0.001) and radiatum layer of CA1
(WT: 8.8 0.3fmol/mg; TG: 10.3 0.2 fmol/mg; p < 0.01).
Conclusions:
At 7 months old there was no difference in spatial memory of WT and TG, but this last group showed a significant reduction in
aversive-related memory. Moreover, we observed a significant increase in 7 nAChR density in some hippocampus areas of this
same group. This is consistent to earlier observations of our research team that showed an increase in 7 nAChR density in some
hippocampal areas as a result of chronic infusion with A in rats, together with a decrease in aversive-related memory. Thus,
another role to this receptor than its participation in memory processes is plausible. It was already shown a nicotinic antiinflammatory pathway, which is dependent on the 7 nAChR. Therefore, this kind of receptor may be playing a compensatory
mechanism, in central nervous system, in order to avoid the neuroinflammatory process associated with the neurodegeneration.
Keywords: alpha7 receptor, Alzheimer's disease, memory
Financial Support: FAPESP, CAPES.
Resumo:23-181
EFFECT OF POSTNATAL FLUOXETINE EXPOSURE ON ANXIETY AND BIOMARKERS OF OXIDATIVE

STRESS IN HIPPOCAMPUS OF FEMALE RATS.
Morais, D. S. C. 1; Silva, A. I. 1; Torres, M. S. 1; Lima, S. 1; Galindo, L. C. M. N. 1; Manhes-de-castro, R.

1
; Lagranha, C. J. 7
1
DEPTO. DE FISIOTERAPIA/ UNIVERSIDADE FEDERAL DE PERNAMBUCO, UFPE
2
DEPARTAMENTO DE NUTRIO/UNIVERSIDADE FEDERAL DE PERNAMBUCO, UFPE
3
DEPARTAMENTO DE ANATOMIA/UNIVERSIDADE FEDERAL DE PERNAMBUCO, UFPE
4
5
DEPARTAMENTO DE ANATOMIA/UNIVERSIDADE FEDERAL DE PERNAMBUCO, UFPE
6
7
NCLEO DE ED. FSICA E CINCIA DO ESPORTE/ CAV-UFPE, CAV/UFPE
Objectives:
The brain is particularly vulnerable to oxidative stress as a result of relatively low levels of antioxidants, high levels of
polyunsaturated fatty acids and/or increased need of oxygen. Numerous studies have confirmed that mood disorders are
characterized by immune system activation and inflammation in the central nervous system, which promotes the production of
reactive oxygen species by several mechanisms. In the treatment of these disorders are used drugs that increase serotonin levels in
brain. In this study we try to relate the chronic effect of inhibition of serotonin reuptake during lactation with the anxiety behavior
and biomarkers of oxidative stress in the hippocampus.
At the first day after weaning the breed were organized with 8 females offspring per dam. Each breed was divided into two
groups: Fluoxetine (GF= 8) and Control (GC= 8). Each group received daily subcutaneous (s.c.) injection of fluoxetine (10
mg/kg, 1l/g, BW) or drug vehicle (sterile 0.9% NaCl 1l/g, BW), respectively, at 7:00 a.m. from the 1st to the 21st postnatal
day. At 55 days of life was assessed anxiety test using the elevated plus-maze following Pellow et al (J Neurosci
Methods.14(3):149-67,1985). At 60 days old of age the animals were anesthetized and the hippocampus was quickly removed
and saved in -80 C until oxidative stress analysis. The hippocampus was homogenized in buffer containing protease inhibitors
and centrifuged at 4500 rpm during 10 minutes at 4C. The supernatant was removed and total protein concentration was
quantified following Bradford protocol (Anal Biochem. 72:248-254,1976). Quantification of catalase was following Aebi
(Methods Enzymol. 105:121,1984) and analysis of MDA was following Draper and Hadley (Methods Enzimol. 186:42131,1990). Chronic treatment with fluoxetine in the neonatal period increased the number of entries (GS = 6.7 +/- 1.7, GF = 10.0
+/- 2.4, p
Conclusions:
These results suggest the existence of a programmer effect in the mechanisms of anxiety in response to alterations in serotonin
concentration during the organization and standardization of brain functions. Our results also suggest that animals subjected to
treatment with selective inhibitors of serotonin reuptake inhibitors, shows increase in protection against free radical formation in
the hippocampus. These lower levels of oxidative stress can be associated with reduction of anxiety even after 40 days of
completion of treatment.
Keywords: ANXIETY, FLUOXETINE, OXIDATIVE STRESS, SEROTONIN
Financial Support: The present study receives financial support from CNPq and FACEPE.
Resumo:23-182
ROLE OF POTASSIUM CHANNELS IN THE ANTIDEPRESSANT-LIKE EFFECT OF FOLIC ACID IN THE

FORCED SWIMMING TEST IN MICE
Ribeiro, C. M. ; Budni, J. ; Freitas, A. E. ; Binfar, R. W. ; Amorim, M. B. ; Rodrigues, A. L. S.

Bioqumica/Universidade Federal de Santa Catarina, UFSC
Objectives:
Potassium channels have been implicated in depressive disorders and in the mechanism of action of antidepressants. Considering
that many studies have indicated that folic acid, a water-soluble B-vitamin, plays a role in the pathophysiology of major
depression, the present study investigated the involvement of potassium channel in the antidepressant-like effect of folic acid. For
this aim, our group investigated the effect of different types of K+ channel inhibitors on the anti-immobility effect of folic acid in
the FST.
Adult Swiss mice of either sex (homogeneously distributed among groups), weighing 3040g were maintained at 21-23C with
free access to water and food, under a 12:12 h light:dark cycle (lights on at 7:00h).The experiments were performed after
approval by the Ethics Committee of the Institution (PP00557). The treatment of mice by intracerebroventricular (i.c.v.) route
with sub-effective doses of glibenclamide (an ATP-sensitive K+ channels inhibitor, 0.5 pg/site), charybdotoxin (a large- and
intermediate-conductance calcium-activated K+ channel inhibitor, 25 pg/site) or apamin (a small-conductance calcium-activated
K+ channel inhibitor, 10 pg/site), augmented the effect of folic acid (10 mg/kg, p.o.) in the FST (16.860.71%, 20.231.58% and
13.050.43% reduction, respectively, when compared with control group 100%). Additionally, the administration of folic acid
and the K+ channel inhibitors, alone or in combination, did not affect locomotion in the open-field test. Moreover, the reduction
in the immobility time elicited by an active dose of folic acid (50 mg/kg, p.o.) in the FST was prevented by the pre-treatment of
mice with a K+ channel opener cromakalim (10 g/site, i.c.v.) (97.415.81%, when compared with control group 100%), without
affecting locomotor activity.
Conclusions:
The results of this study indicate that the antidepressant-like effect of folic acid in the FST may be dependent of its modulatory
effects on neuronal excitability, via inhibition of K+ channels.
Keywords: Folic acid, Potassium channel, Depression, Forced swimming test, Antidepressant
Financial Support: CAPES, CNPq and FINEP-IBN-Net
Resumo:23-183
THE AVERSIVE AND ANTINOCICEPTIVE EFFECTS INDUCED BY NITRIC OXIDE ARE ATTENUATED BY THE
BLOCKADE OF CORTICOTROPIN-RELEASING FACTOR TYPE 1 RECEPTOR WITHIN THE
PERIAQUEDUCTAL GRAY OF MICE.
Miguel, T. T. ; Nunes-de-souza, R. L.
Farmacologia - Faculdade de Cincias Farmacuticas, UNESP/Araraquara
Objectives:
The periaqueductal gray matter (PAG) is a midbrain site markedly involved in fear/anxiety-evoked responses as well as in
nociception. The atypical neurotransmitter nitric oxide (NO) has been related to the modulation of defensive reactions when
released in limbic brain structures, including the PAG. Besides NO, also the neuropeptide corticotropin releasing factor (CRF)
plays a role in the modulation of defensive responses and nociception. The present study investigated whether the anxiogenic and
antinociceptive effects produced by intra-PAG injection of a NO donor, NOC-9 (6-(2-hydroxy-1-methyl-2-nitrosohydrasino)-Nmethyl-1-hexanamine), are attenuated by the blockade of CRF type 1 receptors with NBI 27914 (5-chloro-4-(Ncyclopropyl)methyl-N-propyl)-2-methyl-6-(2,4,6-trichlorophenyl) aminopiridine), within this midbrain structure.
Swiss male mice received surgical implantation of stainless steel guide cannulae intra-PAG. At testing, each mouse received
combined intra-PAG injections of the CRF1 antagonist, NBI (0 or 2 nmoles/0.2 l), and, ten minutes later, NOC-9 (N9, 0 or 75
nmol/0.2 l). Immediately after intra-PAG injection procedure, animals were individually placed in an arena for a period of 5
minutes when the following behaviors were recorded: frequency of jumps (J), time (in seconds) of running (R), freezing (F),
locomotion (L) and frequency of rearing (RE). Forty-eight hours later, a similar procedure was carried out, except that the
animals were injected with formalin (2.5%) into the right hind paw (nociceptive stimulus) and 15 min later, each mouse received
combined intra-PAG injections as described above. Nociception (N) was measured by recording the time (in seconds) spent
licking the paw injected with formalin for a 10-min period. All results were initially submitted to Levene's test for homogeneity
of variance, being, if significant, transformed to log, square root or cube root before being submitted to two-way analysis of
variance (ANOVA) (factor 1: pretreatment and factor 2: treatment) followed by post hoc Duncan test. In those cases (R,J and F)
where Levenes test remained significant after transformations, data were analyzed by Kruskal-Wallis non-parametric followed
by Dunn test. In all cases, a P value 0.05 was required for significance. Results confirmed preliminary study showing that intraPAG NOC-9 (veh+N9) produced aversive-like behaviors (J, R, F) and decreased exploratory behaviors (L and R). Intra-PAG
injection of NBI antagonized almost all aversive effects (J and R) and attenuated the antinociceptive effect induced by NOC-9
(p< 0.05). Results are summarized as follow: veh+veh: 0 (J, R and F), 128.916.7 (N); veh+N9: 5.31.9 (J), 13.45.0 (R), 9328
(F), 34.610.9 (N); NBI+N9: 0.550.55 (J), 1.540.8 (R), 25.510 (F), 107.618.9 (N). Regarding to the exploratory behaviors
(L and RE), intra-PAG NOC reduced both behaviors. Intra-PAG NBI, however, blocked only the reduction of locomotion
induced by NOC-9 (veh+veh: 81.39.9, veh+N9: 34.110.9, NBI+N9: 75.411).
Conclusions:
Intra-PAG NOC-9 was able to induce aversive responses and antinociception and decreased exploratory behaviors. Prior intraPAG NBI 27914 prevented most of the NOC-9 effects. These results suggest that the aversive-related behaviors elicited by NO
release within the PAG may be mediated by CRF action at CRF1 receptors located in this limbic midbrain structure.
Keywords: antinociception, corticotropin-releasing factor, nitric oxide, defensive behaviors, periaqueductal gray
Financial Support: CAPES, CNPq, Fapesp
Resumo:23-184
CANNABIDIOL, CB1R AND CB2R ANTAGONISTS, REVERT THE LOCOMOTOR SENSITIZATION INDUCED BY
ETHANOL IN MICE
Filev, R. 1; de Pauli, R1; Coelhoso, C. C. 1; da Silveira, D. X. 1; dos Santos Jg Jr2,1; Mello, L. E. 1

1
Fisiologia/ Universidade Federal de So Paulo, UNIFESP
2
Farmacologia/ Faculdade de Ciencias Mdicas Santa Casa SP, FCMSCSP
Objectives:
Neuronal plasticity in mesolimbic pathways plays an important role in drug addiction, including alcoholism. Moreover, the
endocannabinoid system has an important modulatory influence over the dopaminergic system. Despite the well established role
of CB1 signaling in drug addiction, recent evidences demonstrate that CB2 receptor is also linked to alcoholism. Here, we
investigate the effects of cannabidiol (CBD) (a phytocannabinoid that increases endocannabinoids levels), as well as, AM251 (a
CB1 receptor antagonist) and AM630 (a CB2 receptor antagonist), alone or in combination with CBD, over the locomotor
sensitization induced by ethanol.
Adult male Swiss mice were daily injected with ethanol (2 g/Kg, i.p. 15 % v/v), for 21 consecutive days. Control mice were
similarly injected with saline. After 7 days abstinence, all animals were challenge with ethanol (1.4 g/kg, i.p.). After the first and
last ethanol injections and after ethanol challenge, the animals were put in an activity cage and their locomotor activity was
measured for 15 min. During the abstinence, the animals received AM251 (1mg/kg, i.p.), AM630 (1mg/kg, i.p.) and CBD
(2.5mg/kg, i.p.). As expected, all experimental groups chronically treated with ethanol developed locomotor sensitization [F(7,
66)=7.2490, p
Conclusions:
Our results suggest that pharmacological manipulation of endocannabinoid system with CBD, as well as with CB1 and CB2
antagonists, alone or in combination with CBD, may potentiate the interaction of endocannabinoids with other cannabinoid
(PPAR) or vanilloid (TRPV1) receptors. These interactions may play a role in the behavioral results encountered in this study.
Keywords: Cannabidiol, Cannabinoids, Ethanol, Locomotor Sensitization, Mice
Resumo:23-185
5-HT1A RECEPTORS IN THE DORSOMEDIAL HYPOTHALAMIC NUCLEUS ARE IMPLICATED IN THE
ANTIPANIC-LIKE EFFECT CAUSED BY CHRONIC FLUOXETINE ADMINISTRATION.
de Bortoli, V. C. 1; Zangrossi, H. Jr. 2

1
Depto de Cincias da Sade, CEUNES - UFES
2
Depto de Farmacologia, FMRP - USP
Objectives:
The electrical or chemical stimulation of the dorsal periaqueductal gray matter (DPAG) or dorsomedial hypothalamic nucleus
(DMH) induces defensive reactions, such as escape behavior, that are suggestive that the experimental animal is undergoing a
markedly aversive experience. Given the striking similarities between the autonomic and behavioral effects of DPAG or DMH
stimulation and the symptoms of panic attacks, it has been suggested that these areas are involved in the genesis of panic disorder
in humans and that their stimulation in animals can model panic attacks. The microinjection of the 5-HT1A receptor agonist 8OH-DPAT or the preferential 5-HT2A receptor agonist DOI into these areas inhibits the escape reaction evoked by local
electrical stimulation. Long-term systemic treatment with antipanic drugs such as imipramine, alprazolam and fluoxetine
facilitates the anti-escape effect caused by the intra-DPAG injection of these 5-HT agonists. This chance has been implicated in
the mode of action of antipanic drugs. In the present study we investigated whether long-term treatment with fluoxetine may also
interfere with the anti-escape effect caused by intra-DMH injection of 8-OH-DPAT or DOI. Using the elevated T-maze (ETM),
we also evaluated whether intra-DMH administration of the 5-HT1A receptor antagonist WAY-100635 blocks the anti-escape
effect caused by fluoxetine.

Male Wistar rats (250-280 g) chronically (21 days) treated with fluoxetine (10 mg/Kg) were intra-DMH injected with 8-OHDPAT (8 nmols), DOI (16 nmols) or saline. The threshold of aversive electrical stimulation that applied to the DMH evokes
escape behavior was measured before and after the microinjection of these agonists. In experiment 2, rats were injected with
fluoxetine (10 mg/Kg) or saline for 20 days. In the next day, ten minutes before the last injection of fluoxetine, the animals were
intra-DMH injected with WAY-100635 (0.37 or 0.74 nmol) or saline. Thirty minutes later they were tested in the ETM. The
effects of intra-DMH injection of drugs were analyzed using a repeated measures ANOVA. Commission ethical protocol n
077/2008 CETEA-FMRP/USP. Results: In rats chronically (i.p.) injected with saline, intra-DMH administration of 8-OHDPAT or DOI significantly raised the threshold of aversive electrical stimulation for inducing escape [ threshold (mean EPM;
A): saline = 6.50 3.77; 8-OH-DPAT = 22.00 1.69 or DOI = 30.50 6.46; p
Conclusions:
5-HT1A receptors in the DMH are implicated in the panicolytic-like effect of fluoxetine, but in contrast with 5-HT2A receptors,
they are not sensitized after chronic administration of this antidepressant.
Keywords: 5-HT1A receptor , dorsomedial hypothalamic nucleus , elevated T-maze, fluoxetine, panic attacks
Financial Support: Fapesp and CNPq, Brazil
Resumo:23-186
QUANTIFICATION OF SYNAPTIC TERMINALS IN HIPPOCAMPUS OF TRANSGENIC MICE FOR
ALZHEIMERS DISEASE SUBMITTED TO ENRICHED ENVIRONMENT STIMULUS.
Balthazar, J. 1; Schowe, N. M. 1; Monteiro-silva, K. C. 2; Caetano, A. L. 2; Buck, H. S. 2; Viel, T. A. 1,3

1
Institute Biomedical Science, USP
2
Department of Physiological Sciences, FCMSCSP
3
School of Arts, Sciences and Humanities, USP
Objectives:
In a recent work, we submitted transgenic mice for Alzheimers disease (TG - PDGFB-APPSwInd) to enriched environment
during 5 months (from 2 to 7 months old). This strain show loss in hippocampal neuronal number and functionality since 2
months of age and senile plaques from 7 months of age. After cognitive stimulation in enriched environment, TG mice showed
better performance in spatial and aversive-related memories, when compared to similar non-stimulated animals. The aim of this
work was to quantify synaptic terminals in the hippocampus of TG mice submitted to enriched environment.
After behavioral observations, animals were killed by decapitation and the brains were extracted and freezed (-80C) until use.
Brain samples (20 m) were obtained in a cryostat and sections were mounted on gelatin-coated slides. Sections were fixed using
acetone 100% and immunostained with antibody against synaptophysin (1:2000) for 6 hours followed by secondary biotinilatedantibody horseradish peroxidase-conjugated IgG for 1 hour. The immune complexes were detected using diamine benzidine
solution. Images were quantified using a MCID image analysis system. In brain samples from stimulated TG animals it was
observed a significant increase (P < 0.05) in synaptophysin labeling in the pyramidal cells (Py) (0.46 0.03, n=11) and in the
radiatum layer (Rad) of CA1 area (0.48 0.02, n=11), when compared to non-stimulated TG samples (0.39 0.01, n=10 and
0.43 0.07, n=14, respectively). Similar observations were made in the granular layer of the dentate gyrus (GrDG) of stimulated
TG mice (0.41 0.02, n=12), when compared to non-stimulated TG animals (0.36 0.01, n=14). Regarding the WT animals, a
significant increase in synaptophysin labeling was observed only in the GrDG of stimulated mice (0.41 0.02, n=13, P < 0.05),
when compared to control mice (0.36 0.01, n=12). No increase in synaptophysin labeling was observed in the polymorphic or
in the molecular layers of the dentate gyrus of stimulated WT or TG mice, when compared to control animals.
Conclusions:
It is well known that exposure of humans or animals to enriched environment increases neuroplasticity as well as cognitive
functions. The TG mice stimulated in enriched environment showed an increase in synaptic terminals in Py and Rad of CA1 and
in the GrDG of hippocampus. Py contains the cell bodies of the pyramidal neurons which are the principal excitatory neurons of
the hippocampus and CA1 area is directly associated with memory formation. GrDG is one of the few regions of the adult brain
where neurogenesis occurs and contains the cell bodies of the dentate gyrus. Besides, GrDG is thought to have an important role
in the formation of new memories aswell. We conclude that the exposure of TG mice to enriched environment improved the
neuronal conexions by means of the increase, at least, in the synaptic number in these areas. This could be associated to the
memory improvement observed earlier.
Keywords: Alzheimer, Enriched Environment, Memory, Synaptophysin
Financial Support: FAPESP, CAPES
Resumo:23-187
EVALUATION OF THE AFFINITY AND INTRINSIC EFFICACY OF LASSBIO-579, AN ORIGINAL NPHENYLPIPERAZINE COMPOUND WITH ATYPICAL ANTIPSYCHOTIC EFFECT
Pompeu, T. E. T. 1; Bettero, G. M. 1; Vieira, R. O. 1; Drummond, C. 1; Fraga, C. A. M. 1; Menegatti, R. 2;

Noel, F. G. 1
1
ICB/ PPGFQM/ Universidade Federal do Rio de Janeiro, UFRJ
2
Faculdade de Farmcia/ Universidade Federal de Goias, UFG
Objectives:
Aim:LASSBio-579 is a N-phenylpiperazine with atypical antipsychotic properties that was obtained by hybridization of
clozapine and a specific D2 ligand (Bioorg. Med. Chem. 11: 4807, 2003) and presented good affinities for the D2 and 5-HT1A
receptors in vitro (Bioorg. Med. Chem. 18:1925, 2010) but a low bioavailability in vivo. As other receptors than the D2 and
5-HT2A could participate in the effect of atypical antipsychotics, the first objective of this study was to assess the affinity of
LASSBio-579 to the D4, 5-HT2C and alpha 2 adrenoreceptors (AR). Our second objective was to estimate its intrinsic efficacy at
the classic aminergic receptors, since the antagonism at D2 and 5-HT2A receptors improves the positive and negatives
symptoms, respectively, whereas agonism at 5-HT1A receptors decreases the liability to induce extrapyramidal symptoms.
Methods: All protocols were approved by the Ethics Committee (CAUAP-UFRJ: DFBC-ICB011). Competition assays: 150 g of
protein obtained from rat cortex (5-HT2A and alpha 2-AR), striatum (D2-like) or hippocampus (5-HT1A) were incubated with
selective radiolabeled ligands, for 15-60 min at 30-37C, in the presence of increasing concentrations of LASSBio-579. The
reaction was terminated by addition of a Tris-HCl (pH 7.4) buffer, followed by rapid filtration and radioactivity counting. For the
D4 and 5-HT2C receptors, we used membrane preparations of cells transfected with human receptors (ChemiscreenTM,
Millipore). Functional Binding: For estimating the intrinsic efficacy of LASSBio-579 at the 5-HT2A and D2 receptors,
competition curves against an antagonist radioligand were performed in the absence or presence of 1 mM GTP (GTP-shift). For
the 5-HT1A receptors, the intrinsic efficacy was estimated by calculating the ratio of the Ki values measured in the presence of an
antagonist and an agonist radioligand (Eur.J. Pharmacol. 386:97, 1999).The data were analyzed by nonlinear regression using
GraphPad Prism 4.0 (USA) to yield the IC50 values that were converted to Ki values by the Cheng- Prusoff equation. Results:
Binding assays: The inhibition curves indicated that LASSBio-579 binds with a lower affinity to the 5-HT2A receptors (Ki= 2.32
M) than to the D2 and 5-HT1A receptors (Ki= 0.10 M). LASSBio-579 also binds with high affinity (Ki=0.14 M) to the D4
receptor, as Clozapine (Ki= 0.064 M). LASSBio-579 has a 16-fold lower affinity for alfa2-AR than Clozapine (Ki= 2.59 M vs
0.16 M) and a 300-fold lower affinity for the 5-HT2C receptors than Clozapine (Ki = 8.0 M and 0.027 M, respectively).
Functional Binding: the inhibition curves in the presence and absence of GTP were superimposed indicating that LASSBio-579 is
an antagonist at these two receptors, just as Clozapine. The assessment of the intrinsic efficacy of LASSBio-579 at the 5-HT1A
receptors is being held.
Conclusions:
LASSBio-579 is a new atypical antipsychotic lead compound, with a sequence of affinities (D2 =D4= 5-HT1A >> 5-HT2A=
alfa2-AR > 5-HT2C) different from Clozapine (5-HT2A/C >>D4>D2 = alfa2-AR > 5-HT1A) albeit sharing the same affinity for
the D2 receptor. As clozapine, LASSBio-579 looks like an antagonist at the 5-HT2A and D2 receptors.
Keywords: Schizophrenia, LASSBio-579, Atypical Antipsychotic
Financial Support: CAPES, CNPq, INCT-INOFAR
Resumo:23-188
DEPRESSIVE-LIKE EFFECT INDUCED BY THE DOPAMINERGIC TOXIN MPP+ IN MICE.
Cunha, M. P. 1; Budni, J. 1; Zomkowski, A. D. E. 1; Amorim, M. B. 1; Ribeiro, C. M. 1; Martins, D. F. 2;

Santos, A. R. S. 2; Rodrigues, A. L. S. 1
1
Depto. de Bioqumica/ CCB, UFSC
2
Depto. de Cincias Fisiolgicas/ CCB, UFSC
Objectives:
Depressive disorders may affect up to 50% of patients with Parkinson disease and are associated with increased disability and
reduced quality of life. Since MPP+ is a toxin used to model Parkinson disease, we investigate the behavioral profile of mice
treated with MPP+ in the tail suspension test (TST) and forced swimming test (FST), and the anhedonic behavior in the splash
test (ST).
C57BL6 male or female mice (3040 g, n = 78) were used. In order to investigate the potential depressant-like effect of MPP+,
mice were treated with MPP+ (1.8-18 g/site, i.c.v.). TST, FST, ST or open-field test (OFT) were carried out 24 h after the
administration of MPP+. In the TST, mice were suspended 50 cm above the oor and immobility time was recorded during a 6
min period. In the FST, animals were individually forced to swim in an open cylindrical container (diameter 10 cm, height 25
cm), containing 19 cm of water at 25C and the immobility time was scored during 6 min. In order to assess the anhedonic
behavior of mice in ST, a 10% sucrose solution was squirted on the dorsal coat of mice and the grooming latency and the total
grooming time during 5 min were registered. The ambulatory behavior was assessed in OFT. The latency to cross the first
quadrant with all paws, the number of crossings, rearings, grooming and fecal boli were counted in a 6-min session. Comparisons
between experimental and control groups were performed by one-way ANOVA followed by Tukey test when appropriate. A
value of P
Conclusions:
These results indicate that MPP+ induces a depressive-like activity in the TST and FST, and an anhedonic like-effect in the ST, in
accordance with epidemiological data that show a comorbidity of Parkinsons disease and depression. These results suggest that
the depressive-like behavior is secondary to the MPP+-induced neurotoxicity.
Keywords: MPP+. , Depressant-Like Effect, Tail Suspension Test, Forced Swimming Test, Splash Test
Financial Support: CAPES, CNPq, FINEP-Rede Instituto Brasileiro de Neurocincia (IBN-Net).
Resumo:23-189
ANTIDEPRESSANTS DECREASE THE EXPRESSION OF PROTEINS INVOLVED IN CELL DEATH IN HUMAN
NEUROBLASTOMA CELLS
Lieberknecht, V. ; Engel, D. ; Mazzucco, S. ; Zomkowski, A. D. E. ; Rodrigues, A. L. S. ; Gabilan, N. H.

Depto. Bioqumica - Universidade Federal de Santa Catarina, UFSC
Objectives:
Mirtazapine and imipramine are antidepressants compounds (AD) which both increase serotonin and noradrenaline
neurotransmission. Some AD show neuroprotective effects, but they also can lead cells to death. The aim of this study was to
explore the effect of AD on the cell viability using human neuroblastoma cells. Also, we investigated if these AD affects the
expression of proteins involved in the cell survival and in the cell death.
Human neuroblastoma cells (SH-SY5Y) were incubated in the presence of 1-100M of mirtazapine (MTZ) or imipramine (IMI)
for 24 and 48h and the cell viability was determined using MTT assay (n=4). To determine the expression of proteins, SH-SY5Y
cells were incubated with MTZ and IMI at concentrations of 2 and 10M for 48h. Then, cells were lysed and total RNA was
extracted. Quantitative real-time PCR (RT-PCR) was performed for Bcl-2, Bax and p53 proteins using specific primers (n=3).
MTZ and IMI (1-20M) did not affect the human neuroblastoma cell viability. The cell viability was reduced by 50M and
100M of IMI to 873% and 561%, respectively. MTZ (100M) reduced the cell viability to 874 %. Human neuroblastoma
cells were incubated with AD at concentrations that they did not cause any cytotoxicity. MTZ (2 and 10M) reduced the
expression of Bax to 596% and 665%, respectively. At the concentration of 10M, MTZ also reduced the p53 expression to
487%. IMI (2M) caused the reduction of Bax and p53 expression to 464% and 426%, respectively. In contrast, IMI (10M)
increased the Bax and p53 expression to 1314% and 1629%, respectively. The treatment of neuroblastoma cells with MTZ (2
and 10M) decreased the expression of Bcl-2 to 745% and 752% respectively and IMI (10M) increased the Bcl-2 expression
to 1479%.
Conclusions:
Mirtazapine and imipramine (AD) induced cytotoxicity on human neuroblastoma cells. At non-cytotoxic concentrations, both
antidepressants reduced the expression of Bax and p53, two proteins involved in the cell death. Our results suggest these effects
may be involved in the neuroprotective action of some antidepressants.
Keywords: ANTIDEPRESSANTS , EXPRESSION , PROTEINS , DEATH , NEUROBLASTOMA
Financial Support: FAPESC, INCT E&N - MCT, CNPq, CAPES-REUNI, PIBIC-UFSC
Resumo:23-190
MTOR PARTICIPATION IN POST-REACTIVATION OF NOVEL OBJECT RECOGNITION MEMORY
Jobim, P. F. C. 1,2,3; Werenicz, A. 1,2,3,5; Maurmann, N. 2; Christoff, R. R. 1,2,3; Pedroso, T. R. 1,2,3; Roesler,
R. 1,2,3,4
1
Lab. de Neurofarmacologia e Biologia de Tumores Neurais, UFRGS
2
Instituto Nacional de Cincia e Tecnologia, INCT-TM
3
Unidade de Experimentao Animal, GPPG-HCPA
4
Laboratrio de Pesquisas em Cncer, CPE-HCPA
5
Objectives:
The formation of memory is a time-dependent process by which an initially labile memory is consolidated into stable long-term
memory (LTM) via de novo RNA and protein synthesis. Considerable evidence indicates that reactivation of a consolidated
memory returns it to a labile state that requires a new time-dependent process to be restabilized (Neuron 36: 521525, 2008), this
process is called reconsolidation, and requires new RNA and protein synthesis. There is evidence that signaling pathways that
control the translation of mRNA are involved in the consolidation and reconsolidation of memories, one of these pathways
related to translational control involves the protein mTOR. The mammalian target of rapamycin (mTOR) is a serinethreonine
protein kinase that acts as a downstream mediator of the PI3K/Akt pathway. Inhibition of mTOR by rapamycin, inhibits the
formation of the complex initiator of translation, an important process for protein synthesis and therefore adversely affect the
consolidation of long-term memory. The aim of this study is to investigate the role of mTOR in post-reactivation of recognition
memory for novel object (NOR) through intrahippocampal administration of rapamycin.
Male Wistar rats were cannulated bilaterally in the CA1 region of dorsal hippocampus. The animals were divided into two
groups: one group received micro infusion of rapamycin, a specific mTOR inhibitor, (600nM/1l into hippocampus); the second
group received vehicle (1% DMSO in saline solution) in the same volumes. At the first day of NOR task, the animals were placed
in the open field box (50x25x40cm) without any object to freely explore the box for 2 minutes. In the second day, animals
returned to the open field box with two identical objects for 5 minutes. In the third day, during reactivation session, the animals
return once again to the open field box, in which one of the objects was replaced by a new object, for additional 5 minutes. The
exploration time of each object was recorded. The infusions took place either 15 minutes before or immediately after reactivation
session. Rapamycin infused into CA1 15 min before reactivation session impaired NOR LTM stabilization pos-reactivation
(P=0.046). Rapamycin infused into CA1 immediately after reactivation session, do not affect NOR LTM (P=0.941).
Conclusions:
Our results suggest that during NOR memory reactivation, mTOR is involved in the synthesis of proteins necessary for the
stabilization of memory in the hippocampus. However, it is possible that the timing of activity of mTOR after NOR LTM
reactivation is short, and for these reason, the rapamycin blocked its effective only when administered 15 minutes before
reactivation session.
Keywords: memory, reconsolidation, mTOR
Financial Support: FIPE/HCPA, CAPES, CNPQ and INCT-TM
Resumo:23-191
ANTIDEPRESSANT-LIKE EFFECT OF CREATINE IN MICE: INVOLVEMENT OF THE SEROTONINERGIC
SYSTEM.
Oliveira, . ; Cunha, M. P. ; Pazini, F. L. ; Machado, D. G. ; Rodrigues, A. L. S.

Bioqumica / Centro de Cincias Biolgicas, UFSC
Objectives:
The relationship between depression and serotoninergic system has been well demonstrated. In this study, we investigated the
involvement of the serotoninergic system in creatine-induced antidepressant-like effect in mice in the tail suspension test (TST), a
predictive test of antidepressant activity.
Swiss male mice (3040 g, n = 711), maintained at 201C with free access to water and food, under a 12:12 h light: dark cycle
were used. In order to investigate the involvement of the serotoninergic system in antidepressant-like effect produced by an oral
administration of creatine, male mice were treated with p-chlorophenylalanine (PCPA, 100 mg/kg, i.p., serotonin synthesis
inhibitor) or vehicle during 4 consecutive days. After 24 hours, they received fluoxetine (10 mg/kg, p.o., serotonin reuptake
inhibitor), creatine (1 mg/kg, p.o.) or vehicle and 60 min later the TST or open-field test were carried out. In order to assess the
behavior of mice in the TST, mice were suspended 50 cm above the oor and immobility time was recorded during a 6 min
period. The ambulatory behavior was assessed in open-field test in a 6 min session. Comparisons between experimental and
control groups were performed by two-way ANOVA followed by Tukey test when appropriate. A value of P
Conclusions:
The results indicate that the serotonergic system (5-HT1A and 5-HT2A receptors and 5-HT reuptake transporter) is involved in
antidepressant-like effect of creatine.
Keywords: Creatine, Antidepressant, Serotonin, Tail Suspension Test, Depression
Financial Support: FINEP-Rede Instituto Brasileiro de Neurocincia (IBN-Net), CNPq and CAPES
Resumo:23-192
IMPORTANCE OF MRNA AND PROTEIN SYNTHESIS FOR RECONSOLIDATION OF AVERSIVE MEMORY
Christoff, R. R. 1,2,3,5; Maurmann, N. 2; Jobim, P. F. C. 1,2,3; Werenicz, A. 1,2,3; Carvalho, L. M. 1,2,3; Pedroso,
T. R. 1,2,3; Roesler, R. 1,2,3,4

1
Lab. de Neurofarmacologia e Biologia de Tumores Neurais, UFRGS
2
Instituto Nacional de Cincia e Tecnologia, INCT-TM
3
Unidade de Experimentao Animal, GPPG-HCPA
4
Laboratrio de Pesquisas em Cncer, CPE-HCPA
5
Pontifcia Universidade catlica do Rio Grande do Sul, PUCRS
Objectives:
New memories are initially formed in a labile state and they need undergo a process of stabilization known as memory
consolidation. Consolidated memories can become sensitive to disruption again after retrieval through its reactivation and it
needs undergo another process of stabilization known as reconsolidation. For many types of memory this processes of
stabilization may differ in molecular mechanisms and brain structures. The aversive memories depend on the recruitment of
molecular mechanisms in the amygdala for the occurrence of memory consolidation and reconsolidation. In both moments of
lability, the process of stabilization requires protein synthesis but the requirement of mRNA synthesis is still a controversial issue.
The aim of this study is to investigate the dependence of synthesis of mRNA and protein in memory reconsolidation of inhibitory
avoidance (IA) in basolateral amygdala (BLA) through the infusion of cycloheximide, an inhibitor of protein synthesis, or DRB,
an inhibitor of mRNA synthesis.
Adult male Wistar rats were cannulated bilaterally in the BLA to permit the administration of the drugs: cicloheximide
(20mg/amygdala); DRB (2mg/amygdala) or vehicle. In IA training, animals learn how to associate a location in the training
apparatus with an aversive stimulus (footshock). On the training trial, rats were placed on the platform and after stepping down
on the grid they received a footshock of 0.7mA for 3s and their latency to step down was measured. The memory reactivation was
carried out 24 hours after learning, the procedure was identical to training except that no footshock was presented. The infusions
of DRB and cycloheximide were performed immediately after reactivation. Administration of cycloheximide also occurred six
hours after the reactivation and without reactivation. The memory retention of the task was accessed through 2 tests, 1 and 10
days later. The post-reactivation infusion of DRB had no effect in the retention of the task while the post-reactivation infusion of
cyclohemide affects the retention of the task in a long-term way as demonstrated in the test 10 days later. Reactivation of memory
is needed for cycloheximide to have an effect, as demonstrated with the infusions without reactivation. The infusion of
cycloheximide 6 hours post-reactivation showed no late effect of cycloheximide in the retention of the task. We used the
Wilcoxon and Mann-Whitney tests to evaluate learning of the task and differences between groups, respectively. Only results
with P-value
Conclusions:
Using cycloheximide and DRB as molecular tools for inhibit protein synthesis and mRNA synthesis our results showed that in
BLA the initial period of memory reconsolidation in IA task requires protein synthesis for memory maintenance for 10 days, but
this protein synthesis is not sustained by de novo mRNA synthesis. These results are in agreement with the hypothesis that the
persistence of memory requires local protein synthesis, possibly in specialized regions of the dendrites of neurons in the BLA.
Keywords: memory, reconsolidation, inhibitory avoidance
Financial Support: CAPES, CNPq, FIPE/HCPA, FAPERGS, INCT-TM
Resumo:23-193
PKC IS INVOLVED IN THE IMPROVEMENT OF MEMORY INDUCED BY SPERMIDINE IN RATS
Guerra, G. P. 1; Rosa, M. M. 1; Ferreira, J. 1,2; Pazini, A. M. 2; Bochi, G. V. 2; Mello, C. F. 2; Rubin, M. A.

1,2
1
Ps-Graduao em Cincias Biolgicas:Bioqumica Toxicolgica, UFSM

2
Programa de Ps-Graduao em Farmacologia, UFSM
Objectives:
Spermidine (SPD) is an endogenous polyamine that modulates N-methyl-D-aspartate (NMDA) receptor function, and has been
reported to facilitate memory formation. The protein kinase C (PKC) signaling pathway seems to play a role in the final phases of
memory consolidation, which require protein synthesis. In the current study we determined whether or not the PKC/CREB
signaling pathway is involved in SPD-induced facilitation of memory of inhibitory avoidance task in adult rats.
All animal experimentation reported in this study was conducted in accordance with the Institutional and National regulations for
animal research (process 0206). Adult male Wistar rats were implanted with 27 gauge cannulas aimed 1 mm above the CA1
region of the hippocampus. After recovery the animals were subjected to a single training session in a step-down inhibitory
avoidance apparatus, consisting of a box with a grid floor and the left portion covered by a platform. The rat was placed gently on
the platform facing the rear left corner. Once the rat stepped down with all 4 paws on the grid, a 3-s, 0.3-mA shock was applied to
the grid. Immediately post-training, PBS, GF 109203X (selective inhibitor of PKC, 0.25 pmol), SPD (0.2 nmol) or GF 109203X
(0.25 pmol) combined with SPD (0.2 nmol) were injected bilaterally into the hippocampus (0.5 L). After the injection, the
animals returned to their home cages. A subset of the animals was sacrificed 30 min after injections and the hippocampus were
dissected for Western blot analysis of PKC (phosphorilated and total PKC) and CREB (phosphorilated and total CREB). The
other animals had a testing session for retention 24 h after training in the inhibitory avoidance apparatus. A step-down latency test
was taken as a measure of retention, with a cut-off time of 600 s. Statistical analysis of step-down latencies during testing
(nonparametric two-way ANOVA) showed a significant SPD or PBS vs GF109203X or PBS interaction [H1 = 6.84, p< 0.05],
indicating that GF 109203X prevents the increase of PKC phosphorilation induced by SPD. [PBS (100.5 12.4); GF109203X
(99.5 18.7); SPD (179 16.5); GF 109203X plus SPD (90 24.7)] (data are expressed as mean SEM of phospho-PKC/total
PKC ratio) Furthermore, the administration of drugs did not alter the total-CREB, phospho-CREB levels and the phosphoCREB/total-CREB ratio.
Conclusions:
These results suggest that SPD-induced facilitation of memory in rats depends on a sequence of biochemical events in the rat
hippocampus that is triggered by PKC phosphorylation, but not by CREB.
Keywords: CREB, Hippocampus, Memory, PKC, Spermidine
Financial Support: CNPq, CAPES, FAPERGS
Resumo:23-194
CANNABIDIOL IMPAIRS THE RECONSOLIDATION OF MORPHINE-ASSOCIATED MEMORY IN RATS
TRAINED FOR CONDITIONED PLACE PREFERENCE
de Carvalho, C. R. ; Cruz, J. S. ; Takahashi, R. N.

Objectives:
Addicts constantly relapse to drug seeking even after years of abstinence and this behavior is frequently evoked by the recall of
memories linked to drug experience. Established memories including appetitive memories can become transiently fragile if
reactivated, during this labile phase, known as reconsolidation. Established memories can be disrupted by interfering of some
pharmacological manipulations. Thus, reactivation of mnemonics traces provides a strategy for disrupting memories that
contribute to pathological states (Trends Neurosci. 28:51, 2005. Considerable evidence demonstrates that endocannabinoid
system is implicated in the regulation of a variety of physiological processes, such as, memory, learning, motivation and drug
reinforcement. Also, it has been suggested that cannabidiol, a nonpsychoactive constituent of cannabis, play a role in the
modulation of addictive behavior (J Neurosci. 29(47):14764, 2009). First we tested whether the reactivation of memory
associated to morphine in CPP evoked by a single conditioning session with morphine remains stable until 2 weeks after
reactivation. Subsequently, we evaluated the effect of post-reactivation treatment with NMDA receptor antagonist MK-801 or
cannabidiol, on the reconsolidation of morphine-related memory, using the same experimental schedule.
Male adult Wistar rats weighing 250-300g at the start of experiment were used. CPP was conducted in a three-compartment
apparatus and the protocol used was modified from Milekic [J Neurosci. 26(11):3010, 2006]. The rats were conditioned for four
consecutive days to 10 mg/kg morphine or saline in an unbiased fashion. One week later, both groups of rats (n=8/group)
received one additional conditioning trial (10 mg/kg of morphine or saline, s.c.) in order to induce the reactivation of morphineCPP memory. The reconsolidation of morphine-CPP memory was tested 24h, 1 week and 2 weeks after reactivation. In the
second experiment, immediately after reactivation, the rats were divided in three groups (N=5-6/group) and received two
systemic injections of MK-801 (0.20 mg/kg, i.p.), cannabidiol (10 mg/kg, i.p.) or vehicle 5 hours apart. Subsequently, all animals
were retested until 2 weeks to determine whether the effect on memory reconsolidation was stable. Only morphine-conditioned
rats and that had recalled morphine-memory spent more time in the drug-paired side compared to respective control group, over
two weeks post reactivating memory (P0.05). These findings suggest that re-experience of a conditioning reinforced session is
necessary to maintain stable morphine-associated memory. Furthermore, similar to the group that received MK-801 post
reactivation cannabidiol-treated group showed a significant decrease in time spent in morphine paired side over time, when
compared with control group (P 0.05).
Conclusions:
Our study shows for the first time that both MK-801 and cannabidiol weaken permanently the motivational memory associated to
morphine in rats trained in CPP. The opportunity of weakening persistent and unwanted memories by disrupting the
reconsolidation process opens new therapeutic perspectives to attenuate seeking behavior and risk to relapse in addicts.
Keywords: addiction, cannabidiol, conditioned place preference, memory reconsolidation, morphine
Resumo:23-195
BLOCKADE OF DEFENSIVE BEHAVIOR AND ANTINOCICEPTION INDUCED BY A PREDATOR WITH INTRAAMYGDALA MIDAZOLAM IN MICE.
Barbalho, C. A. ; Canto-de-souza, A.
Universidade Federal de So Carlos, UFSCar
Objectives:
Aim: The antinociception induced by threatening situations has been suggested as part of the defensive repertoire of rodents
(Behav. Brain. Sci., 3; 291, 1980). In this context, we have shown that confinement of mice into the open arm of the elevated plus
maze, a widely used animal model of anxiety, produces antinociception sensitive to intra-amygdala injection of midazolam, a
benzodiazepine receptor agonist (Psychopharmacol., 150; 300, 2000). Previous findings have demonstrated that mice display
defensive behavior when exposed to a predator (a rat) (Physiol. Behav. 81; 465, 2004). The present study investigated (i) whether
the exposure of mice to a rat also elicits antinociception in the prey and (ii) the effects of intra-amygdala injection of midazolam
on this type of pain inhibition.
Methods and Results: Swiss male mice (n= 6-8/group) received bilateral intra-amygdala injection of midazolam (0, 3.0 or 30
nmol/0.1 l) and 5 minutes later they were injected with 0.6% acetic acid i.p. (0.1 ml/10g; nociceptive stimulus). Then, each
mouse was placed in the rat exposure test (RET), which consists of a rectangular wooden box divided into 3 compartments: the
accommodation of the live rat or toy rat (A-side), separated from the mouse exploration area (B-side) by a wire mesh and
protected chamber of the mouse (C-side), separated from the exploration area by small door. All mice were individually placed
on the B-side and were allowed to explore freely both B-side and C-side of the box for a period of 10 minutes. The following
measures were recorded: total number of contortions (TC) and number of contortions displayed in the unprotected area (CB-side),
time spent and frequency of entries into the B-side (TB-side and EB-side), and risk assessment [total stretched attend postures
(TSAP), protected SAP (pSAP), time of contact with the grid (GRID). Two-way ANOVA (condition x treatment) revealed
significant effects (p , 0.05) for condition [TC: F(1,35)= 5.00; TB-side: F(1,35)= 5.46; EB-side: F(1,35)= 5.89; pSAP: F(1,35)=
12.61], treatment [TC: F(2,35)= 9.21; TB-side: F(2,35)= 78.70; EB-side: F(2,35)= 9.14; TSAP: F(2,35)= 32.02; pSAP: F(2,35)=
5.39; GRID: F(2,35)= 306.74] and condition x treatment interaction [TC: F(2,35)= 4.48; TB-side: F(2,35)= 9.36; TSAP: F(2,35)=
3.78; pSAP: F(2,35)= 6.29; GRID: F(2,35)= 4.29]. Post hoc analysis showed that the predator-prey model induced
antinociception in mice treated with saline and exposed to the live rat. Intra-amygdala midazolam (3.0 and 30 nmol) increased the
number of contortions in mice exposed to live rat and increased the TB-side, EB-side and time of contact with the grid.
Conclusions:
Conclusion: This study showed that mice exposed to the RET display antinociception, corroborating previous studies that have
demonstrated pain inhibition in mice exposed to anxiety test (Psychopharmacol. 150; 300, 2000). Intra-amygdala midazolam
attenuated the both antinociception and defensive behaviors elicited by the predator. These results suggest that
GABA/benzodiazepine mechanisms located within the amygdala play a role in both defensive behavior and antinociception in
mice exposed to a predator.
Keywords: AMYGDALA , ANTINOCICEPTION , MICE, PREDATOR , RET
Financial Support: UFSCar, CAPES
Resumo:23-196
EFFECTS OF INTRA-PERIAQUEDUCTAL GRAY INJECTION OF MIDAZOLAM ON MEMORY ACQUISITION
AND ON ANXIETY IN MICE
Pereira, B. C. ; Nuncianto, A. C. ; Baptista, D. ; Fachini, G. ; Canto de Souza, A.

Universidade Federal de So Carlos, UFSCar
Objectives:
Several studies have shown that benzodiazepines (BDZ) produce anxiolytic-like effects in different animal models of anxiety. In
addition, BDZ drugs also impair learning and memory performance in rodents. We have previously shown that the aversive
stimulus (footshock) facilitates memory expression in the Step-down test (SD) and elicits anxiety in the elevated plus-maze
(EPM). In this context, intra-periaqueductal gray (PAG) injection of midazolam partially reverses both footshock effects,
suggesting that GABAA receptors modulate these behavioral responses (SBNeC, J.097, 2010). This study investigated whether
these intra-PAG effects of midazolam are upon memory acquisition on anxiety-like state.
Swiss male mice (25-30g, n=7-9/group) received intra-PAG injection of midazolam (MDZ: 0 or 30 nmol/0.1 l) and were placed
on the platform of the step-down test to record the latency (in seconds) to step down (L1). Half of animals treated with MDZ (0
or 30 nmol) received an electric footshock (FS; 0,5mA) for 10 s while another half did not (without FS). Twenty-four hours later,
each mouse received a second intra-PAG injection of saline or MDZ and were exposed once again to the step-down test to record
the latency to step-down (L2) in a condition without shock. The following groups were formed: [sal + sal; sal + MDZ; MDZ +
sal; MDZ + MDZ]. Step-down latency on the test day (L2) was recorded as an index of inhibitory avoidance memory and
anxiety. t-test for independent samples showed that intra-PAG MDZ reduced the latency to step down compared saline group in
L1 [t(59)= 2.19, p
Conclusions:
The present results suggest that exposure of mice to SD (shock - aversive stimulus) facilitated the aversive memory, characterized
by increase in L2. Intra-PAG midazolam produced an anxiolytic-like effect characterized by a decrease in L2
(saline+midazolam). However, intra-PAG MDZ impaired acquisition of aversive memories in mice. Taken together, these results
suggest that GABAA receptors located within the PAG seem to modulate both memory and anxiety responses.
Keywords: Anxiety, EPM, Midazolam, PAG, Step Down
Financial Support: UFSCar, CAPES
Resumo:23-197
SHORT-TERM CHOLECYSTOKININ-4 INTRAPERITONEAL TREATMENT ENHANCES AMPHETAMINEINDUCED STEREOTYPED BEHAVIOR IN RATS.
Fischer, B. N. 1; Vergamini, L. B. 1; Silva, H. R. 1; Lopes, C. 1,2; Tieppo, C. A. 1,3

2
Faculdade da Sade/ Universidade Metodista de So Paulo, UMESP
1
Cincias Fisiolgicas/Fac. de C. Mdicas da Santa Casa - SP, FCMSCSP
3
Faculdade de Cincias Humanas e da Sade - PUC - SP, PUC-SP
Objectives:
Cholecystokinin, the most abundant neuropeptide in CNS, shows many effects related to dopaminergic behaviors and plasticity.
Previous results reveal that acute CCK4 (CCK2 receptor agonist) and CCK8S (non-specific CCK agonist) icv injections increase
amphetamine-induced stereotyped behavior. The goal was to evaluate the effects of short-term CCK4 and CCK8S ip treatments
on late amphetamine-induced stereotypy.
Forty male Wistar rats were divided in four groups and during seven days (once a day) received one of these ip treatments: CCK4
(6,0 nmol/kg), CCK8S (6,0 nmol/kg), amphetamine (10 mg/kg) or saline (1 ml/kg). Before a seven day abstinence period, all
groups received an amphetamine injection (10 mg/kg), the stereotyped behavior was quantified for 30 min and stereotypy was
immediately evaluated by Setler scale (10 s were observed in every 10 min, during 30 min). The animals under CCK4 treatment
showed stereotypy scores of 5.0 at a 20 minute observation period {5.0;9.0}, higher than control (4.0 {2.0;6.0}) and CCK8S (4.5
{2.0;6.0}) groups. The amphetamine treated rats (8.0 {6.0;11.0}) displayed a more intense stereotyped behavior than the other
three groups. The Mann-Whitney test was used.
Conclusions:
Amphetamine treatment produced enhanced scores compatible to well described amphetamine sensitization. The CCK4
treatment, which is more specific to stimulate CCK2 receptors response, also provokes augmented sensitivity to amphetamine but
less intensively than amphetamine. The results suggest that the seven day CCK4 ip treatment induces plastic changes on
cathecolaminergic systems and CCK2 receptors are more likely to be involved in these effects.
Keywords: dopamine, cholecystocinin, amphetamine, stereotypy, CCK-4
Financial Support: Fundao Arnaldo Vieira de Carvalho
Resumo:23-198
ANTIOXIDANT MODULATION ANTIDEPRESSANT-LIKE EFFECTS OF RESVERATROL IN MICE
Moritz, B. ; Schmitz, A. E. ; Souza, L. F. ; Ucha, M. F. ; Dafre, A. L.

Depto. de Bioqumica/Universidade Federal de Santa Catarina, UFSC
Objectives:
Resveratrol (3,5,4' trihydroxy-trans-stilbene) is a phytoalexin produced by some plants displaying several biological actions, such
as antitumoral, anticancer along with his known antioxidant activity. The oxidative stress seems to be a common characteristic of
several pathologies, including mood disorders. Acute resveratrol treatment decreases monoamine reuptake and monoamine
oxidase A and B activity in rat sinaptossomes, which was associated to its antidepressant-like action. These data suggest that
resveratrol can be used in the treatment of depression.
In the present study, animals were treated with different doses of resveratrol (0.1 20 mg/kg) by oral route for 21 days. After
that, the forced swimming test (FST) and the tail suspension test (TST) were performed to assess antidepressant-like action, while
the open field test (OFT) tested the locomotor activity. Glutathione reductase (GR), glutathione peroxidase (GPx) and glyoxalase
(GLO) were measured in cortex, hippocampus and cerebellum of mice. Resveratrol decreased the immobility in the FST and in
the TST, without interfering with locomotion. Resveratrol decreased the GR and GPx activities in the cortex, hippocampus and
cerebellum.
Conclusions:
These data suggest that resveratrol has antidepressant-like effect in mice, but, contrary to what was expected, 21 days resveratrol
treatment decreased the GR and GPx activity, reinforcing the need for further experiments to determine the mechanism of action
of resveratrol.
Keywords: Antidepressant-like, Antioxidant, Resveratrol
Financial Support: CNPq, FAPESC, INCT-ET
Resumo:23-199
PURINERGIC RECEPTOR BLOCKADE INDUCES ANTIDEPRESSANT- AND ANTICOMPULSIVE-LIKE
EFFECTS
Pereira, V. S. 1; Sato, V. A. H. 1; Casarotto, P. C. 1; Sartim, A. G. 2; Guimares, F. S. 1; Joca, S. R. L. 2

1
Farmacologia, FMRP-USP
2
Fsica e Qumica, FCFRP-USP
Objectives:
The activation of P2 receptors (P2R) by ATP leads to a transient increase in calcium influx at presynaptic neuronal cell and
increases glutamate release [1]. The activation of post-synaptic P2R is speculated to activate nitric oxide (NO) synthesis in the
brain [2]. Interestingly the glutamatergic and nitrergic systems are strongly involved in several stress-related psychiatric disorders
[3]. Thus, the aim of the present study was to investigate the effects of a P2R antagonist administration to animals submitted to
models predictive of antidepressant-, anxiolytic- and anticompulsive-like effects.
Male Swiss mice (weighting 25-35g) were injected with PPADS (6.25, 12.5, 25 or 50 mg/kg), imipramine (30 mg/kg), paroxetine
(5 mg/kg) or vehicle and submitted to forced swimming test (FST), elevated plus maze (EPM) or marble burying test (MBT) 30
min after the drug injection. Data was analyzed by one-way ANOVA followed by Dunnet's post hoc test when appropriated.
Animals receiving PPADS (12.5mg/kg) or imipramine (30mg/kg) presented reduced immobility time in the FST [F(5,32)=11.85;
p0.05 vehicle: 35.2+/-4.8; PPADS 12.5mg: 39.1+/-6.1; 25mg: 35.3+/-5.9; 50mg: 42.5+/-4.0; Mean+/-SEM of percent time spent
in open arm] and no locomotor activity alterations [F(3,17)=0.87; p>0.05; vehicle: 11.3+/-0.8; PPADS 12.5mg: 11.8+/-1.5;
25mg: 9.8+/-1.0; 50mg: 11.8+/-0.4; Mean+/-SEM of enclosed arm entries] were found.
Conclusions:
These results suggest that purinergic neurotransmission mediated by P2R could modulate behavioral responses associated to
stress-related psychiatric disorders and could be a potential useful approach to understand the neurobiology and treatment for
these disorders. The specific type of P2R able to mediates these behavioral effects is only speculative due to the nonspecific
action of PPADS over P2XR or P2YR. Modulation of glutamate release and NO production seems to be the main way that
purinergic transmission exerts its behavioral effects. Thus, further studies are needed to verify if this findings could be
reproduced in other models and if it is correlated with alterations in glutamate release and NO production elicited by purinergic
blockade. [1] Neuroreport 21:865, 2010. [2] Prog Neurobiol 84:40, 2008. [3] Pharmacol Rev 58:58, 2006.
Keywords: Anticompulsive, Antidepressant, ATP, Purinergic system , Stress
Financial Support: FAPESP, CNPQ, FAEPA
Resumo:23-200
EVALUATION OF THE ANTINOCICEPTIVE ACTIVITY OF THE THIOPHENIC 6CN10 DERIVED
Penha, A. R. S. 1; Mota, C. D. A. X. 1; Maia, A. K. D. H. L. 1; Oliveira, A. M. D. 1; Mota, V. G. 1; Almeida,

R. N. D. 1; Moura, R. O. D. 2; Assis, T. S. D. 1; Diniz, M. D. F. F. D. M. 1; Morais, L. C. S. L. D. 1
1
UNIVERSIDADE FEDERAL DA PARABA, UFPB
2
UNIVERSIDADE ESTADUAL DA PARABA, UEPB
Objectives:
The discovery of new drugs with activity in the central nervous system, as powerful painkillers and anti-inflammatory drugs,
returned to millions of people a quality of life that seemed lost forever, thanks to the effort of scientists who work in this area.
Thus, the purpose of this study was to investigate the possible antinociceptive effects of thiophene-derived 6CN10 in animal
models using rodents.
The following tests were performed: writhing induced by acetic acid, formalin test and hot plate test. The protocols were
approved by the ethics committee on animal research, under the number 0501/11. Swiss male mice, 25-35 grams, were divided
into groups of eight animals. The control group received distilled water and tween 20; 3 experimental groups received the test
substance at doses of 25, 50 and 100mg/kg and the standard group received morphine (10mg/Kg). All administrations were
performed intraperitoneally. In the writhing test induced by acetic acid, the total number of contortions was recorded for 10
minutes. At all doses used of 6CN10, we observed a significant reduction in the number of contortions, (25: 2.10 0.6; 50: 5.2
1.3 and 100: 0.4 0.3) compared to the control group (30.8 2.8). In the formalin test, which the standard substance was injected
into the sub plantar region of mice, the time licking the paw was used as indicative of nociceptive response. It was found that
there was a significant decrease in paw licking time during the first phase in animals treated with the derivative 6CN10 (25: 69.5
7.0; 50: 83.9 6.0 and 100: 60.9 6.4) compared with the control group (116 3.0). The same occurred in the second phase, at
all doses of 6CN10 (25: 49 17, 50: 28 12.5, 100: 10.2 0.9) compared to the control group (158 40.0). The hot plate test
quantifies the animal's reaction time to thermal stimulus, ie the time when the animal is placed on a hot plate at 52.0 0.5 C to
lick a paw or present the behavior of lifting (or jumping). In animals treated with 6CN10 at the doses tested, there was a
significant increase in the latency to perception of thermal stimulation at a dose of 50mg/kg at 30min (50: 13.5 2.5). 60 min
after, there was a significant increase of latency at doses of 50 and 100mg/kg (50: 12.5 6.3; 100: 10.4 2.0) compared to the
negative control group at 30 min (5.0 0.8) and 60 min (5.3 0.7), respectively. The results were submitted to statistical analysis
using the unpaired Student t test (p <0.05).
Conclusions:
Analysing the results obteined, we suggest that the isolated thiophenic 6CN10 derived, using the via and doses tested, showed an
antinociceptive activity demonstrated through in vivo methodologies for analgesia. Further studies are necessary in order to
elucidate the mechanisms involved in the observed effects.
Keywords: thiophenic, derived, antinociceptive
Financial Support: CAPES, UFPB
Resumo:23-201
ENVIRONMENTAL ENRICHMENT DURING ADOLESCENCE PROTECTS EMOTIONAL, OLFACTORY AND
COGNITIVE DEFICITS INDUCED BY RESERPINE IN RATS
Frana, S. L. ; Souza, R. R. ; Bessa, M. M. ; Takahashi, R. N.

Objectives:
Emotional, cognitive and olfactory abnormalities are generally described in Parkinsons disease (PD) and may precede motor
impairments. Since the early diagnosis and intervention can be decisive for patients prognosis, animal models of nonmotor
deficits are important to characterize early symptoms and develop neuroprotective therapies. So far, there is no available effective
strategy that modify PDs outcome. However, several studies have demonstrated beneficial effects promoted by environmental
enrichment (EE) in animal models of neurodegenerative process through cognitive, visual and motor stimulus. Therefore, the
objective of this work was to assess whether the treatment with low doses of reserpine is able to induce olfactory, emotional and
cognitive deficits that resemble PDs symptoms in rats, as well as to evaluate a possible preventive effect of EE over these
impairments.
Reserpine-induced (1.0 mg/kg, ip., 24 hours prior test) emotional, cognitive, olfactory and motor impairments were evaluated in
adult male Wistar rats. Emotional effects of reserpine treatment were assessed in the Forced Swim Test (FST).The FST consists
of submitting the animals for 5 minutes in a cylinder filled with water, andthe immobility time (s) induced by this inescapable
situation was recorded. The animals treated with reserpine showed significant increase in immobility time (25.8 3.9 X 46.7
6.8; p= 0.01). Olfactory and cognitive deficits were evaluated in the Olfactory Discrimination Test (ODT) and in the Olfactory
Fear Conditioning (OFC). The ODT consists of allowing rats to freely investigate a cage divided into two identical
compartments: one with fresh sawdust (unfamiliar) and another with familiar sawdust (from its home cage). The time spent in the
familiar compartment was considered an olfactory discrimination index. The animals treated with reserpine showed a significant
time decrease in the familiar compartment (55.4 5.0 X 38.5 5.9; p=0.04).The OFC consists of conditioning a neutral odor
(CS) with an unconditioned stimulus (US: 5 footshocks, 0.5 mA/2s). The association between the CS + US induces defensive
behaviors (DB), which is evaluated by exposing the rats to the previously paired CS. Reserpine induced deficits in DB such as
hide time (75.9 3.4 X 42.9 11.1; p= 0.04).These impairments were independent from motor or anxiogenic-like effects,
measured in open field test. Additionally, we assessed the effects of 10 weeks exposure of rats to the EE on the reserpine-induced
impairment. The reserpine treatment did not affect rats exposed to EE, once there was no significant difference compared to their
respective EE control (p>0.05). The results indicated that EE exposure prevented the emotional, olfactory and cognitive deficits
induced by reserpine treatment.All procedures used in the present study were submitted to the UFSC Ethics Committee on the
Use of Animals (PP280), which follows the Principles of laboratory animal care from NIH.
Conclusions:
These results suggest that acute administration of low doses of reserpine in rats could be a useful tool to investigate nonmotor
symptoms of PD. Moreover, exposure to EE may prevent changes induced by reserpine in rats, suggesting that stimulus during
early life promotes behavioral changes that can act as neuroprotective factor.
Keywords: Parkinsons disease, Reserpine, Environmental enrichment, nonmotor symptons, neuroprotection
Financial Support: Capes, CNPq, FAPESC and UFSC.
Resumo:23-202
ACUTE EFFECT OF CAFFEINE ON ENDOCANNABINOID-INDUCED MEMORY IMPAIRMENT
Moreira Silva, D. M. ; Campos-lima, T. ; Marin, M. T. ; Santos, B. R.

ICBIM/Universidade Federal de Uberlndia, UFU
Objectives:
The formation of a new memory depends on several molecular events and connections among the neural circuits. It starts with the
acquisition of information, followed by a consolidation process and, finally, the retrieval of stored memories. Neural circuits
related to memory may still be influenced by many substances, as caffeine, which has neuroprotective function and improves
cognitive processes, as well as anandamide, an endocanabinoid, present at high levels in brain areas involved to mnemonic
system. The aim of this study was to evaluate the effect of anandamide on memory acquisition and consolidation and how
caffeine affects these processes.
Methods:Memory acquisition and consolidation were evaluated at Elevated Plus Maze (EPM). Male swiss adult mice were
placed on the end of one of the open arms, facing away from the center, and the time taken by the animal to enter one of the
closed arms in the day 1 (transfer latency 1 (TL1)) was recorded with the help of a stop watch. Then, 24 hours later, transfer
latency was recorded again (TL2). Generally, memory process induces a decrease of TL2 related to TL1. First, it was evaluated
the effect of anandamide and caffeine on memory acquisition. In this test saline (NaCl 0.9%) or caffeine (10 mg/kg) was
intraperitoneally (i.p.) administered 30 min before TL1 and saline or anandamide (1 mg/kg) was i.p. administered 15 min before
TL1. Then, there were 4 experimental groups: saline+saline, saline+anandamide, caffeine+saline and caffeine+anandamide.
Later, it was evaluated the effect of anandamide and caffeine on memory consolidation. In this test saline or caffeine was i.p.
administered 15 min after TL1 and saline or anandamide 30 min after LT1. In both experiments TL2 was recorded in the absence
of drug administrations. Results were expressed as meanS.E.M of the time (seconds) of Transfer Latency and analyzed by twoway ANOVA followed by Bonferroni post-hoc test (p
Conclusions:
Anandamide impaired memory consolidation process, but not the acquisition. Moreover, acute caffeine had significant
neuroprotective action preventing anandamide-induced memory consolidation impairment. Nevertheless, there is still necessity of
more studies to clarify the molecular mechanisms and neural systems involved in this process
Keywords: Memory, Caffeine, Endocannabinoid, Acute
Financial Support: Federal University of Uberlndia
Resumo:23-203
EXERCISE PRE-CONDITIONING REDUCES BRAIN INFLAMMATION, PROTECTS AGAINST BRAIN-BLOODBARRIER BREAKDOWN AND MOTOR FUNCTION IMPAIRMENT INDUCED BY TRAUMATIC BRAIN INJURY
Mota, B. C. ; Rodrigues, F. S. ; Gerbatin, R. R. ; Stamm, D. N. ; Della-pace, I. D. ; Ferreira, A. P. O. ;

Souza, M. A. ; Silva, L. F. A. ; Royes, L. F. F.
Objectives:
Traumatic brain injury (TBI) is a major cause of disability and death in young adults that could be present neuronal cell loss,
microglial activation, and intraparenchymal hemorrhage in cortical regions. In this context, the early inflammatory response after
brain injury induces leukocytes influx to lesion site leading edema and breakdown of the blood-brain barrier (BBB). Indeed,
studies indicated that treatment whit drugs that antagonize endogenous inflammatory mediators reduce post-injury edema and
improve motor function. Although it is believed that physical exercise on neurorehabilitation after traumatic brain injury may be
useful, little information is available regarding the prophylactic role of physical exercise on deleterious effects induced by TBI.
Therefore, we decided to investigate whether previous physical training prevents against events as acute inflammatory response
(IL-1; IL-6; IL-10 TNF-), BBB breakdown and motor function impairment induced by FPI.
Male Wistar rats, 90-day-old (250-300g) were used in this study. The physical training was carried during four weeks on
treadmill. At the end of training, sedentary and trained rats were implanted with a cannula on cortex. Next day, the animals were
underwent to fluid percussion brain injury (FPI) or not (SHAM). Twenty four hours later motor function was evaluated and
cytokine levels (IL-1; IL-6; IL-10 and TNF-), fluorescein plasma extravasation (indicator of a BBB breakdown) were measured.
The FPI induced the increase of IL-1 [F(2,39)=20,35; pF(2,39)=9,05; pF(2,39)=2,22; p=0.803], and FPI decreased IL-10
[F(2,39)=16,14; pper se [F(2,39)= 36,41; pF(2,39)= 2,41; pF(2,39)=5,49; pF(2,39)=11,00; pF(1,13)=22.56; pH(3)=22.73;
p<0.001].
Conclusions:
Considering that inflammatory events together with cytotoxic effects of immune mediators lead to injured brain, physical training
may be a new therapeutic approach to control acute inflammation that lead to long-term cell damage and neurobehavioral
disability after TBI.
Keywords: Brain-Blood-Barrier, Inflammation, Motor Function, Physical Exercise, Traumatic Brain Injury
Financial Support: CAPES, CNPq
Resumo:23-204
INFLAMMATION EXARCEBATES MEMORY IMPAIRMENT INDUCED BY GLUTARIC ACID PUPS RATS
Rodrigues, ; Mota, ; Gerbatin, ; Satmm, ; Della-pace, ; Ferreia, ; Souza, ; Fighera,

DMTD/Universidade Federal de Santa Maria, UFSM
Objectives:
Glutaric academia I (GA-I), an inherited deficiency of glutaryl-coenzyme A dehydrogenase characterized by accumulation of
glutaric acid and neurological symptoms. It has been reported that patients with GA-I have cognitive impairment after
encephalopathic crises, which are precipitated by infectious processes during an age-dependent susceptibility period. How
neurological crises are typically precipitated by common infection and febrile illness, this indicates a potentiating role of
inflammatory cytokines in neurological disorders this organic acidemia. The objective was to evaluate the performance of pups
rats injected chronically with GA, in the absence and presence of LPS, in a test of spatial memory. Moreover, evaluate the
content of cytokines and possible strutuctural alteration on hippocampus.
Were used Wistar pups rats, with four days age (5 30g). Vehicle (saline 0.9%) or buffered GA, pH 7.4 (5 mol g of body
weight1) was administered subcutaneously, twice per day, from the 5th to the 28th day of life to produce brain concentrations of
GA of around 0.6 mol g1, ~0.72 mM, similar to concentrations those found in glutaric acidemic patients. . The pups rats were
injected intraperitoneally (i.p.) with lipopolysaccharide (LPS 2 mg/kg; E.coli 055 B5) or vehicle, once per day, from 25th to 28th
day of life to mimic an infections state. Cytokine levels (IL-1&beta and TNF-&alpha) were measured using a ELISA Kit. The
Cavalieri method was used to estimate the volume of the hippocampus. Barnes maze was used to assessment of spatial learning
and memory in rodents. At the second day of test, statistical analysis demonstrated a learning deficit in GA-Veh, Sal-LPS and
GA-LPS [F(3,43)=8.82; PF(3,43)=4.56; PF(1,35)=9.70; PF(1,37)=8.44; PF(1,12)=28.13; P
Conclusions:
GA-chronic treatment caused a deficit of spatial learning in pups rats, potentiated by the presence of an inflammatory process.
Thus, given the high degree of limitation that GA-I brings to children with this disease, understanding of mechanisms involved in
neurological disorders induced by accumulation of organic acid, is important for the development of new therapies to treat this
condition.
Keywords: Cytokines, Glutaric Acidemia, Memory, Neuroinflamation
Resumo:23-205
PROCONVULSANT ACTIVITY OF THE MEROPENEM ON PILOCARPINE-INDUCED SEIZURES MODEL IN
MICE
Lima, C. N. C. ; Linhares, M. I. ; Rios, E. R. V. ; Venncio, E. T. ; Lopes, K. S. ; Carvalho, A. M. R. ;

Nascimento, N. L. ; Martins, A. M. C. ; Fonteles, M. M. D. F. ; Rodrigues, F. T. S. ; Oliveira, G. V. D.
department of pharmacology and Physiology, UFC
Objectives:
To investigate the acute effect of Meropenem (MERO) in model of seizures induced by pilocarpine (P400) in mice through
behavioral observation and measurement of amino acids in homogenates of two brain areas (hippocampus -HC; striatum-CE).
Swiss mice (28-33g) were pretreated with MERO (500 and 1000 mg/kg, iv, n = 8 each) and after 10 min the seizures were
induced with P400 mg/kg, ip. The control group was treated only with P400. Behavioral assessment was carried out for 60 min,
following the parameters: latency to first convulsion and latency to death. For amino acids determination, HPLC was used. The
results were presented as mean S.E.M. Data were analyzed by ANOVA and Student-Newman-Keuls as post hoc, with
significance of p
Conclusions:
Our findings showed proconvulsant effects of Meropenem and we suggest that the probable mechanism involved can be related
with increase in the amount of excitatory amino acids and reduction in the amount of inhibitory amino acids, thereby promoting
excitotoxicity, but this mechanism should be further investigated.
Keywords: Meropenem, Pilocarpine, seizure
Financial Support: CNPq and CAPEs
Resumo:23-206
PROTECTIVE EFFECT OF ANACARDIC ACID OBSERVED IN BEHAVIORAL ASSESSMENTS OF
EXPERIMENTAL MODELS OF HUNTINGTON'S DISEASE
Freitas Neto, H. P. ; Medeiros-linard, C. F. B. ; Sereniki, A. ; Amorim, A. F. C. ; Pereira, R. C. R. ; Silva, S.

N. ; Lafayette, S. S. L.
Depto de Fisiologia e Farmacologia/ Univ Federal Pernambuco, UFPE
Objectives:
Huntington's disease (HD) is a progressive neurodegenerative disorder characterized by involuntary movements choreiform,
emotional disturbances and dementia. In HD there is a selective loss of GABAergic neurons in the striatum, caudate nucleus and
putamen, and a generalized atrophy of the cortex. Several evidences have shown that excessive production of free radicals is
strongly associated with the development of many diseases typical of aging. The antioxidant activity of extracts of cashew
(Anacardium occidentale L.) has been attributed to the presence of phenolic compounds, particularly acid anacardic. A liquid
made from cashew nut is one of the richest sources of acid anacardic. The main aim of the study was to investigate the
neuroprotective effect of acid anacardic analyzing behavioral parameters of experimental models of HD.
Were utilized 3 groups of 8 animals each (Male Wistar rats), weighing 280g to 350g and maintained under controlled
illumination and temperature. Was administered for 8 days, intraperitoneally, saline in the animals of first group (G1) and 3nitropropionic acid (3NP), 20mg/kg, in the other two groups (G2 and G3). The 3NP inhibits the enzyme succinate
dehydrogenase, causing oxidative stress and neuron death, resulting in changes similar to DH. In the same period, the animals of
G1 and G2 received water by gavage (10mL/kg), G3 received anacardic acid (50mg/kg). On the first day after treatment was
done in the open-field behavioral test, it was observed for 5 minutes the frequency of locomotion (number of floor units entered
by the animal), rearing frequency (the number of times the animal stood on its hind legs) and time of resting (time that the animal
stood still without showing any movements). Then the animals were subjected to the elevated plus maze test, to assess the
percentage of memory retention, and the rota-rod test that assesses motor skills and balance by the length of stay on the rotating
shaft. Results were analyzed by ANOVA followed by Tukey test and values were expressed as mean S.E.M. (p
Conclusions:
The results showed that animals treated with 3-nitropropionic acid presented motor and Cognitive deficits, and daily
administration of acid anacardic, 50 mg/kg, attenuated the neurobehavioral changes, showing that this compound arises as a
potential neuroprotective agent and stimulating further research.
Keywords: ANACARDIC ACID, HUNTINGTON'S DISEASE, NEURODEGENERATIVE DISEASES,
NEUROPROTECTION
Financial Support: FACEPE, CAPES, CNPq.
Resumo:23-207
BED NUCLEUS OF THE STRIA TERMINALIS ADRENOCEPTORS ARE INVOLVED WITH EXPRESSION OF
CONTEXTUAL FEAR CONDITIONING
Hott, S. C. ; Gomes, F. V. ; Fabri, ; Reis, ; Correa, ; Resstel,

Department of Pharmacology, School of Medicine , USP
Objectives:
The bed nucleus of the stria terminalis (BNST) is a limbic structure associated with autonomic, neuroendocrine and behavioral
functions which seems to be critically involved in the expression of anxiety-like responses as contextual fear conditioning.
Among the numerous neural inputs to the BNST, noradrenergic synaptic terminals are prominent and some evidence suggests an
activation of this BNST neurotransmission during aversive situations. Thus, the aim of this work was to study a the involvement
of BNST noradrenergic system on the modulation of the behavioral and autonomic responses induced by contextual fear
conditioning.
Male Wistar rats (240-270g) with cannulae implanted bilaterally into the BNST were submitted to a 10 min conditioning session
(6 footshocks, 1.5 mA, 3 s). Twenty-four h later, the behavioral and autonomic responses, mean arterial pressure (MAP) and
heart rate (HR), evoked by aversive context were measured during test session (test) for 10 min. It was administered 10 min
before test 0.1L of saline (n=6), 15 or 25 nmol of a non-selective -adrenergic receptor antagonist propranolol (n=5), 10 or 20
nmol of a non-selective -adrenergic receptor antagonist fentolamine (n=6) or the combination of both drugs (n=6). : Compared
to control animals, both dose of propranolol significantly reduced the percentage of the freezing (F(2,15)= 5.561; P<0.001).
Conclusions:
Our findings showed that noradrenergic neurotransmission within the BNST, through adrenergic receptors activation, is involved
in the expression of responses induced by contextual fear conditioning.
Keywords: autonomic responses, adrenoceptors, bed nucleus of the stria terminalis, contextual fear conditioning, noradrenergic
system
Financial Support: FAPESP, CNPq, Capes and FAEPA.
Resumo:24-073
PARADOXICAL INCREASE IN LIVER KETOGENESIS DURING LONG-TERM INSULIN-INDUCED
HYPOGLYCEMIA IN DIABETIC RATS
Barrena, H. C. ; Schiavon, F. P. M. ; Gazola, V. A. F. G. ; Furlan, M. M. D. P. ; Bazotte, R. B.

Pharmacology and Therapeutics / State University of Maring, UEM
Objectives:
In certain physiological conditions including fasting, pregnancy, hyperlipidic diet and prolonged exercise, a modest ketonemia
elevation seems to be important as an energy source to all extrahepatic tissues including the brain. Conversely, diabetic
ketoacidosis is a potentially life-threatening complication that happens predominantly in type 1 diabetes because these patients
could show a clinic hyperketonemia posthypoglycemia. It is well established that insulin inhibits liver ketogenesis. However,
during insulin-induced hypoglycemia (IIH) the release of counterregulatory hormones could overcome the insulin effect on
ketogenesis. To clarify this question the ketogenic activity in livers from alloxan-diabetic rats submitted to long-term IIH was
investigated. Moreover, liver glycogenolysis, gluconeogenesis, ureagenesis and the production of L-lactate were measured, and
its correlation with blood levels of ketone bodies (KB), L-lactate, glucose, urea and ammonia was investigated.
We conducted experiments in male Wistar rats (Rattus norvegiccus). For this purpose, overnight fasted alloxan-diabetic rats
(DBT group) were compared with control non-diabetic rats (NDBT group). Diabetes (DBT group) was induced by an intravenous
injection of alloxan (40 mg/kg dissolved in saline). Long-term IIH was obtained with an intraperitoneal injection of Detemir
insulin (1 U/kg), and KB, glucose, L-lactate, ammonia and urea were evaluated at 0, 2, 4, 6, 8 or 10 hours after insulin injection.
Because IIH was well established two hours after insulin injection this time was used for liver perfusion experiments. Therefore,
at this time the rats were anesthetized with intraperitoneal sodium thiopental (40 mg/kg). After laparotomy, a cannula was
inserted into the portal vein for in situ liver perfusion. The perfusion fluid was Krebs-Henseleit (KH) bicarbonate buffer. In
addition to glucose, in part of the experiments the liver production of L-lactate, urea and pyruvate were also evaluated. The basal
rates of ketogenesis and the rates of ketogenesis during the infusion of octanoate (0.3 mmol/L) were evaluated. The results were
reported as means standard deviation (SD) of six to eight individual experiments. The administration of Detemir insulin
decreased (P < 0.05) blood KB and glucose levels (n=8), but there was an increase in the blood L-lactate, urea and ammonia
levels and a rebound increase in blood KB during the glucose recovery phase of IIH (n=8). In agreement with these results, the
capacity to produce KB from octanoate was increased in the livers of DBT rats (n=6). Moreover, the elevated blood L-lactate
levels in DBT rats (n=8) could be attributed to the higher (P < 0.05) glycogenolysis when part of glucose from glycogenolysis
enters glycolysis, producing L-lactate. In contrast, except glycerol, gluconeogenesis was negligible in the livers of DBT rats
(n=7).
Conclusions:
During long-term IIH the higher liver ketogenic capacity of DBT rats increased the risk of hyperketonemia. In addition, in spite
of the fact that the insulin injection decreased blood KB, there was a risk of worsening lactic acidosis.
Keywords: ketogenesis, hypoglycemia, experimental diabetes, Detemir insulin
Financial Support: CNPQ and Fundao Araucria
Resumo:24-074
PROTEIN FRACTIONING OF BREAST MILK SAMPLES USING BIODIMENSIONAL ELECTROPHORESIS (2D
PAGE).
Santos, F. A. D. ; Machado, A. U. ; Braga, C. P. ; Neves, R. C. ; Pozzi, C. M. C. ; Lima, P. M. D. ; Moraes,

P. M. D. ; Tsutomunakatani, B. ; Padilha, P. D. M.
Qumica e Biqumica/Instituto de Biocincias, UNESP
Objectives:
Improve methods of biodimensional electrophoresis (2D PAGE) for the fractioning of proteins from samples of milk.

Breast milk samples were treated with hexane for five hours to extract lipid fraction. Subsequently, 250 mL was collected from
protein fraction and the proteins were precipitated with acetone in 1:4 ratio. After this procedure, the precipitated protein was
resolubilized in a buffer containing urea, thiourea, CHAPS, ampholytic, blue of bromophenol and DTT. The extract containing
resolubilized proteins was applied in isoelectric focusing strips of 13 cm, with precast gel containing immobilized ampholytes of
pH 3-10 and pH 4-7. These strips were rehydrated during 12 h and then were taken to the IEF system to run in first dimension.
The average time of this step was 4.5 h. In second dimension overrun, strips were equilibrated in equilibrium buffer with SDS,
Tris-HCl pH 8.8, urea, glycerol, blue of bromophenol, DTT and iodoacetamide for 30 min. Then the tapes were applied on
polyacrylamide gels of 12.5% and 10% (w / v). A sheet of filter paper was then placed on the polyacrylamide gel, beside the
strip, to which was applied 10 L of standard proteins with molar mass in the range of 14 97 kDa. The strips and filter paper
containing the standard proteins were then sealed in the polyacrylamide gel with 0.5% agarose (w / v). The average time of
separation in second dimension was 12 hours. The races of the gels at 12.5% (w / v) with pH 3-10 and 10% (w / v) and with pH
4-7 were performed in triplicate. After separation, the proteins in gel were fixed and revealed by colloidal Coomassie staining.
The resulting gels were then scanned using a GE Healthcare scanner. The scanned images were analyzed to determine the
correlation between the gel repetitions and to make spot counts, using the ImageMaster platinum 7.0 software program. Gels
image analysis of electrophoretic overrun showed a large diversity of proteins with molecular weight between 14.40 and 97.00
kDa, highlighting some proteins of molecular weight between 30.00 and 97.00 kDa which showed higher expression. Overall,
there was a good resolution between gels repetitions; however, in pH 4.7 of 10% (w / v) gel, the fractioning of proteins with
molecular weight between 33.00 to 97.00 kDa occurred more efficiently. The correlation analysis indicated a mean number of
296.4 and 151.19 protein spots, which represent 80% and 71% of protein were present in repeated gels of pH 3 -10 and 4-7,
respectively.
Conclusions:
The electrophoretic procedure in 2D PAGE gels allowed a protein fractioning more efficient when used the gradient of pH 4 -7 in
gel concentration of 10% (w / v). When was used a gradient of pH 3-10 with concentration of 12% (w/v), inspite of obtaining a
larger number of protein spots, the proteins fractioning was not as efficient as compared to a gradient of pH 4 - 7.
Keywords: Human milk, 2D PAGE, Electrophoresis, Protein Fractioning
Resumo:24-075
MATERNAL OBESITY: IMPLICATIONS FOR MOTHER AND OFFSPRING
Ornellas, F. P. C. ; Souza-mello, V. ; Costa, R. N. L. ; Mandarim-de-lacerda, C. A. ; Aguila, M. B.

DEPARTAMENTO DE ANATOMIA , UERJ
Objectives:
Maternal obesity is increasingly prevalent and may affect the health of the child. We investigated the effects of maternal dietinduced obesity in mice on male and female offspring biochemical and biometrical parameters.
Female C57BL/6 mice were fed either a standard chow (SC, 17% energy from fat) or a high-fat diet (HF, 49% energy from fat)
for 8 weeks before mating and throughout pregnancy and lactation. Mating occurred at 3 months-old. Body weight of mothers
was evaluated daily and Oral Glucose Tolerance Test (OGTT) was performed before breeding. Sacrifice of dams occurred after
weaning. Offspring of SC and HF dams were weaned onto standard chow and studied at 3 months-age. Body mass of offspring
was measured weekly. In all cases the significance level of P<0.02).
Conclusions:
These data indicate that maternal high-fat diet administered prior and during gestation and lactation, programs carbohydrate
metabolism and obesity in both male and female offspring.
Keywords: HIGH FAT DIET, MATERNAL OBESITY, FETAL PROGRAMMING, C57BL/6
Resumo:24-076
NEONATAL MALNUTRITION AND BACTEREMIA IN PROCESS OF DENTAL ALVEOLITIS IN RATS:
BACTERIOLOGICAL AND HISTOLOGICAL ANALYSIS
Arajo, F. R. G. D. 1; Sampaio, B. 1; Costa, T. B. 1; Morais, N. G. 1; Alcntara, G. A. L. F. 1; de Castro, C.

M. M. B. 2
1
Federal University of Pernambuco, Laboratory of Immuno
2
Federal University of Pernambuco, DN
Objectives:
to study the process of dental alveolitis in its bacteriological and histological aspects, relating them to the development of
bacteremia in undernourished adult rats in the neonatal period.
We used 40 Wistar male rats, suckled by their mothers, fed a diet during lactation containing 17% casein in the nourished group
(N) or 8% casein in the undernourished group (UN). After weaning, the animals were given the Labina standard diet, containing
23 % mixed protein until the end of the experiment. After 90 days, the animals underwent upper right incisor extraction,
induction of alveolitis and divided randomly into four groups: N-21, UN-21, N-28 and UN-28. The oral microbiota was obtained
using a swab soaked in 40L of 0.9% sterile NaCl and blood culture through one hundred microliters of venous blood collected
from the tail of each animal. These procedures were performed before the extraction, 5 minutes after extraction, on the 21st day
after the alveolitis for groups N-21 and UN-21 and on the 28th day after the alveolitis for groups N-28 and UN-28. After 21 or 28
days of alveolitis the levels of osteocalcin and ionized calcium were determined by chemiluminescence method and calculated
method, respectively. To perform the quantification of these levels 5 ml of blood was collected from each animal by cardiac
puncture. In order to obtain the histological findings, the dental socket was divided into three thirds: apical, middle and cervical.
In each of these, the stage of healing was examined by an image analysis system consisting of an optical microscope coupled to a
Motic microcamera and microcomputer containing the Motic Image Plus software. This allowed the measurement in micrometers
of the following histological structures: bone tissue, connective tissue, blood clot and inflammatory tissue. Data were expressed
as mean standard deviation. The t test was used for parametric data and the Mann-Whitney test for nonparametric data.
Statistical significance was considered by assuming a critical level of 5%, which yielded the following results: before and after
extraction a lower bacterial growth was observed per colony forming unit (CFU) in the peri-alveolar region of the upper right
incisors of undernourished animals, while the opposite was true after alveolitis, when a larger number of CFU was seen in these
animals. The percentage of positive blood cultures obtained after the alveolitis was higher in the undernourished animals.
Regarding to osteocalcin, a reduction was observed in group N-28 and an increase in group UN-28; the availability of ionized
calcium was higher in the nourished animals and the alveolar healing process was also more development in these animals.
Conclusions:
The neonatal malnutrition was able to cause a decrease in the immune response of the animals, promoting noticeable changes in
their defenses against infectious agents, even in the adult organism. These changes were seen in the composition of the
microbiota from the peri-alveolar region and in the development of bacteremia, thereby demonstrating the existence of a direct
relationship between oral and systemic diseases. Moreover, there was a delay in the alveolar healing process with changes in the
levels of osteocalcin and ionized calcium.
Keywords: MALNUTRITION, BACTEREMIA, DENTAL ALVEOLITIS, PERI-ALVEOLAR MICROBIOTA, ALVEOLAR
HEALING
Financial Support: CAPES/PROPESQ
Resumo:24-077
EFFECTS OF MATERNAL INTAKE OF PALM OIL, DURING PREGNANCY AND LACTATION, ON ADULT
OFFSPRING ADIPOSITY
Bispo, K. P. ; Oliveira, L. R. ; Souza, . D. S. S. D. ; Albuquerque, K. T. D. ; Sardinha, F. L. D. C.

Instituto de Nutrio Josu de Castro, INJC
Objectives:
The concept of metabolic programming refers to the capacity of early adaptations to induce permanent metabolic changes and
thereby influence physiologic processes in later life, even after withdrawal of the adverse stimulus. Cardiovascular disease,
insulin resistance with type II diabetes, and energy balance disorders with hyperphagia and obesity are conditions that have been
associated with exposure to different fatty acids. Food industry has been used palm oil as a substitute for trans fatty acids sources.
Influences of palm oil intake during perinatal period are controversial. Although predominant fatty acid is saturated, palm oil has
high oleic acid (monounsaturated) level, possibly, these explain positive results observed in a few number of studies. This study
was designed to observe the effects of intake of palm oil during pregnancy and lactation on body weight gain, adiposity and
ingestion, of offspring at 90 days of life.
Female Wistar rats received isocaloric and normolipidic diet containing palm oil or soybean oil during pregnancy and lactation.
Upon weaning, soybean pups continued on soybean diet (SG) and palm oil offspring (PG) switched to a soybean diet until 90th
days of life. Body weight and food intake was evaluated daily. The 90-day-old animals were decapitated and adipose tissues
(epididymal, retroperitoneal and mesenteric) were removed and weighted. Test T was used to assess differences between groups
using software Prism 5.0 version. As results, SG group (25,950,55) had higher (PG: 23,080,46) food intake at weaning. At 90
days, no difference was observed in body weight (SG:444,210,01; PG:441,114,37) between the groups, although adipose
tissue weight was increased in PG (PG: 18,831,71; SG: 14,750,95).
Conclusions:
Maternal intake of palm oil elevates the quantity of adipose tissue in adult offspring.
Keywords: diet, metabolic programming, palm oil
Financial Support: FAPERJ
Resumo:24-078
PROTEIN RESTRICTION IN INTRAUTERINE LIFE DIFFERENTLY AFFECTS LIPID METABOLISM AND BODY
GROWTH IN FEMALE AND MALE RATS NEWLY WEANED.
Mendona, L. A. C. ; Assis, T. ; Pinheiro, D. F. ; Vicentini-paulino, M. L. M; Denise Sartori

Fisiologia/Instituto de Biocincias, IBB- UNESP- Botucatu
Objectives:
The fetal development is dependent on maternal nutrients and the absence or deficiency of nutrients can directly affect the normal
development, resulting in physiological and metabolic disorders. These changes determine the genesis of several diseases in
adulthood, including obesity, diabetes and hypertension. However, it is unclear if protein deficiency experienced by the mother
during pregnancy can result in short-term changes in lipid metabolism of offspring. Moreover, it is unclear whether the effects of
protein restriction are expressed differently in males and females. The aim of this study was to evaluate the influence of protein
malnutrition in the prenatal period on the body weight (BW), fat deposition in liver, retroperitoneal and perigonadal adipose
tissues and plasma cholesterol in male and female rats at 21 d of age (at weaning).
Female Wistar rats, following confirmation of pregnancy, were divided into two experimental groups that differed in their diet
until the birth of offspring: Group C (control, n = 8) - animals fed a normal protein diet (17% PB) and Group R (restricted, n = 8)
- animals fed a low protein diet (6% PB). During lactation, the mother rats were fed with normal diet. One female and one male
pup from each of the dams were sacrificed at weaning, by decapitation, and blood was collected for determination of plasma
cholesterol. After laparotomy, the liver and retroperitoneal (RP) and perigonadal (PG) adipose deposits were removed and
weighed, and the liver was frozen for later determination of fat. Students t test was used for comparisons between R and C
groups and the significant differences were defined as P
Conclusions:
The results show that protein restriction during intrauterine life promotes changes in body size and lipid metabolism disorders,
which are already significant in the early life of the animal, but that affect males and females differently.
Keywords: intrauterine malnutrition, lipid metabolism, protein restriction
Resumo:24-079
ASSOCIATION BETWEEN PERINATAL FACTORS, BODY MASS INDEX AND EATING DISORDERS IN
COLLEGE STUDENTS
Lofrano-prado, M. C. 1; Amorim, A. C. R. 2; Silva, L. M. L. 2; Prado, W. L. 3; Albuquerque, A. A. U. 2;

Silva, A. L. 2; Lopes-de-souza, S. 4
1
1Programa de Ps-graduao em Nutrio, UFPE
2
Departamento de Nutrio, UFPE
3
Escola Superior de Educao Fsica, UPE
4
Departamento de Anatomia, UFPE
Objectives:
To describe the prevalence of eating disorders symptoms and to verify the associations between perinatal factors (birth weight,
time of lactation and obstetric complications), body mass index (BMI) with symptoms of eating disorders in young college
students (healthy sciences students).
A cross-sectional study was conducted with 358 college students (239 girls and 119 boys), aged between 18 to 23 years old.
Weight and height were measured at students college, and BMI was then calculated (body mass/height2), students were
encouraged to ask to their parents the birth weight, time of lactation and if they presented some obstetric complications (selfreport). Binge eating symptoms was assessed by Binge Eating Scale (BES); to assay the presence of purging was used Bulimic
Investigatory Test Edinburgh (BITE) and Eating Attitude Test (EAT) to anorexia nervosa symptoms. Data are expressed as
relative frequencies, and comparisons between boys and girls were made by chi-square. This study was formally approved by the
ethical committee of the Federal University of Pernambuco (CEP/CCS/UFPE No 143/10). Informed consent was obtained from
all subjects.
Conclusions:
The prevalence of eating disorders symptoms is higher in female than in male college students, and in both genders these
symptoms presented a positive association with BMI. Our data suggests that as higher as BMI, higher is the risk to healthy
science college students to develop eating disorders. In addition, longitudinal studies are needed to better elucidate the
associations between perinatal factors and eating disorders.
Keywords: Eating Disorders, Eating Behavior, Binge Eating, Perinatal Factors
Resumo:24-080
EFFECT OF HYDROLYZED HQSOLABI FROM CHENOPODIUM QUINOA WILLD ON THE HEPATOCYTES
MORPHOLOGY OF WISTAR RATS.
Veloso, N. C. ; Carrara, M. A. ; Stefanello, T. F. ; Bernardo, Z. H. ; Teixeira, C. J. ; Natali, M. R. M. ;

Batista, M. R.
Universidade Estadual de Maring, UEM
Objectives:
To evaluate the effects of hydrolyzed HQ-Solabi from Chenopodium quinoa Willd on the hepatocytes morphology of Wistar
rats.
64 adult Wistar rats (60 days) were divided in four groups: sedentary control group (SCG), sedentary supplemented group
(SSG), exercise control group (ECG) and exercise supplemented group (ESG). During the 30 days of treatment, SSG and ESG
groups received a daily intragastric dose of 2 mL hydrolyzed HQ-Solabi 250 mg/mL distilled water), and SCG and ECG
groups 2mL of saline (0,9%). At this time, ECG and ESG groups were submitted to forced swimming. After the supplementation
period and/or exercise, the livers of these animals were removed and frozen for later morphological analysis of hepatocytes. 10
m thickness semi-serial sections were made on a glass razor using a cryostat, stained by Harris hematoxylin and eosin (HE)
technique and PAS (Periodic Acid-Schiff) histochemistry. The images were captured with the help of a high-resolution camera
coupled to microscope and morphometry performed by an image analysis system. The morphometric analysis showed no
statistically differences between the groups. The presence of inflammatory infiltrates was more common in the ECG and ESG
groups.
Conclusions:
Daily supplementation with hydrolyzed HQ-Solabi at the dose used did not cause changes in hepatocytes morphology and
glycogen intracellular deposition. However, the intense physical exercise protocol applied caused a qualitative increase of the
inflammatory infiltrates in the liver of animals.
Keywords: Chenopodium quinoa Willd, Hepatic morphology, PAS histochemistry, Rats
Financial Support: Solabi do Brasil
Resumo:24-081
EFFECT OF KETOGENIC DIETS WITH DIFFERENT COMPOSITIONS OF POLYUNSATURATED FATTY ACIDS
IN SERUM BIOCHEMICAL PARAMETERS AND PROTEIN S100B IN DIFFERENT NEURAL STRUCTURES OF
RATS
Vizuete, A. F. K. ; Souza, D. F. D. ; Batassini, C. ; Dutra, M. F. ; Costa, A. P. ; Bernardi, C. ; Gonalves, C.

A. S.
Bioqumica- Instituto Cincias Bsicas da Sade, UFRGS
Objectives:
ketogenic diet (KD) bas been use as anti-epileptic treatment in cases of children who resist to antiepileptic drugs. During the
intake of the diet, adaptations occur in the peripheral and nervous metabolism, as the synthesis of ketone bodies in the liver and
the partial replacement of glucose by ketone bodies as an energy source in the brain. However, the action mechanism of
ketogenesis and neuroprotection in epilepsy is not yet understood, and there is some discussion of side effects of this diet on lipid
body profile and cardiovascular system. The KD is composed of: hight concentration of lipidis, low or no carbohydrates and low
protein.This study aimed to standardize ketogenic diets with different compositions of polyunsaturated fatty acids and study their
effects on peripheral and nervous metabolism in order to understand its mechanism of action and their different effects on the
cardiovascular system.
For this purpose, ten Wistar rats (thirty days old) were subjected to different diets: 1) standard diet (SD); 2) ketogenic diet (KD +
SD); 3) ketogenic diet with increased omega-3 composition (KD + O). After eight weeks of treatment, the rats were fasted for 8
hours and sacrificed for perform biochemical markers of serum and S100B protein (ELISA) in cerebrospinal fluid (CSF), serum
and brain structures hippocampus, temporal cortex and striatum. The data obtained are expressed as mean S.E.M.. The
statistical analysis used was one-way ANOVA followed by Tukey's post hoc test (p
Conclusions:
There was no difference in peripheral metabolism between the types of KD. However, was different in some biochemical
parameters when compared to SD. S100 B a calcium- binding protein secreted by astrocytes, in striatum there was difference of
S100B contents between the two types KD maybe this can be occur by a neuroprotective effect of the own ketogenic and/ or
immunomodulation and antiinflammatory activity from the linolenic acid. This work is ongoing and aims to better analyze the
neuroprotective action of the ketogenic diet on the central nervous system.
Keywords: ketogenic diet, PUFAs, epilepsy, S100B
Financial Support: CNPq, CAPES, IBN-Net, FAPERGS
Resumo:24-082
EFFECT OF CHRONIC TAURINE TREATMENT ON BODY WEIGHT AND ON FOOD AND WATER INTAKE IN
NON-OBESE DIABETIC AND NON-DIABETIC RATS
Caletti, G. 1; Pedrollo, E. F. 3; Barros, H. M. T. 1,3; Gomez, R. 2,1

1
Programa de Ps-Graduao de Cincias da Sade, UFCSPA
2
Departamento de Farmacologia - ICBS - UFRGS, UFRGS
3
Disciplina de Farmacologia, UFCSPA
Objectives:
Taurine is one of the most abundant free amino acids in the central nervous system and in the peripheral tissues. Physiological
and therapeutic properties of this amino acid has been explored and revealed that taurine modulates a variety of biological
function, showing antioxidant, osmoregulator, neuromodulator, hypoglycemic, and anti-obesity properties. Anti-obesity property
or changes on food and water intake after taurine administration have been tested only in obese rats. However there are no studies
in non-obese diabetic and non-diabetic rats. Thus, our objective here was to verify the effect of chronic taurine treatment on body
weight and on food and water intake in non-obese diabetic and non-diabetic rats.
Eighty adult male Wistar rats were divided in control (CTR) and diabetes (STZ) group. Diabetes was induced by a dose of
streptozotocin, 60mg/kg, via ip. After diabetes confirmation they were randomly allocated to receive daily 0, 25, 50 and 100
mg/kg of taurine (n=10/subgroup), i.p., for 28 days. During the experimental period water and food intake were measured every
day. Body weight was measured twice a week. Data were grouped and a Two Way ANOVA was used to detect differences
between groups and treatments, followed by the Bonferroni test when appropriate. Our results showed that STZ rats gain less
weight than CTR rats during the experimental period (gain of weight: CTR0 = 22.2 2.3, STZ0: 7.94.9g; P< 0.001). Taurine
did not change the food intake in CTR rats (CTR0: 22.40.7; CTR25: 21.40.9; CTR50: 22.60.5; CTR100: 22.21.3 g/day/rat).
STZ rats drink water more than three times than CTR rats (CTR0: 35.10.7 x STZ0*: 112.52.3 mL/day/rat, *P<0.001).
Conclusions:
Here we did not observe an anti-obesity effect of taurine in non-obese STZ and CTR rats. In CTR rats, chronic taurine treatment
decreases water intake, that is explained on bases on the GABAergic effect of taurine, since GABAA agonists produces a doserelated inhibition of water consumption that are abolished by pretreatment with GABAA receptor antagonists in rats. Chronic
taurine treatment decreases time- and dose-dependently the food and water intake in STZ rats. The highest dose of taurine also
decreases the loss of weight in STZ rats. Because these effects, we hypothesize the taurine increases the energy recovery in
diabetic individuals and may be an alternative drug to delay long-term macrovascular, microvascular, and neurological
complications that are causes of major morbidity and mortality in diabetic patients.
Keywords: obesity, amino acid, appetite, streptozotocin, energetic drinks
Financial Support: CAPES, CNPq, UFCSPA
Resumo:24-083
PHARMACOLOGICAL MANIPULATION OF THE SEROTONINERGIC SYSTEM INDUCES
HYPORESPONSIVITY OF THE CONTROL MECHANISM OF THE SACIETY.
Silva, A. I. 1; Torres, M. S. 2; D'alcantara, S. C. N. L. 2; Novaes, L. C. M. G. 2; Almeida, L. C. A. 2; Sousa,

S. L. 1,2
1
NUTRIO-UNIVERSIDADE FEDERAL DE PERNAMBUCO, UFPE
2
ANATOMIA, UNIVERSIDADE FEDERAL DE PERNAMBUCO, UFPE
Objectives:
Diseases related to energy metabolism such as obesity become a problem of public health worldwide, because it represents one of
the most important risk factor for development of cardiovascular disease and type II diabetes. Many studies have linked obesity to
a nutritional status in early periods of life as a result of an apparent deficit in the mechanism of homeostatic control. The
serotoninergic system plays a crucial role on the development and to the homeostatic and hedonic control of eating behavior.
Under homeostatic control serotonin acts in hypothalamic nuclei stimulating satiety through blocking neurons that produce
orexigenic peptides and stimulates neurons which produce anorexigenic peptides. Changes in the functioning of serotoninergic
neurons during critical periods of development reflect on the ability of the brain and body to adapt and survive. The aims of the
study were verify the effects of neonatal chronic pharmacological manipulation of the serotoninergic system during the
development on changes in eating behavior. Fluoxetine inhibits the reuptake of serotonin from the synaptic cleft and was used in
this study to manipulate the system serotonergic neurotransmission.
In the present study we used male Wistar rats. At the first day after weaning the breed were organized with 8 offspring per dam.
Each breed was divided into two groups: Neonatal Fluoxetine (Fn= 18) and Neonatal Saline (Sn= 18). Each group received daily
subcutaneous injection of fluoxetine (10 mg/kg, 1l/g, BW) or drug vehicle (sterile 0.9% NaCl 1l/g, BW), respectively, at 7:00
a.m. from the 1st to the 21st postnatal day. At 35 days of life was assessed food intake (g) during 24 hours. At 40 days of life was
measured body weight (g) and food intake (g) after 3 hours of fasting. At 41 days was performed to test food intake (g) and
Behavioral Sequence of Satiety (BSS) (s) following Halford e Blundell (Physiol Behav.60(3):933-9,1996) 1 hour after applying
acute dose of fluoxetine (10 mg / kg, 1l/g, BW) (Sn+Fag and Fn+Fag). All data were analyzed by Student t test. Related to body
weight we observe that in Fn group, body weight was lower (p = 0.0001) than Sn group (Sn = 141 +/- 2,29; Fn = 123,94 +/2,02). No difference was observed between groups when measured food intake during 24h (Sn = 16,1 +/- 0,41; Fn = 16,8 +/- 0,3,
p = 0,1736) and after three hours of fasting (Sn = 3,01 +/- 0,17; Fn = 3,36 +/- 0,22, p = 0,2220). However, after receiving acute
dose of fluoxetine Sn+Fag showed a reduction in food consumption when compared to its own control (Sn =3,01 +/- 0,17;
Sn+Fag = 2,31 +/- 0,21; p = 0, 0142) and the Sn+Fag showed no significant difference (Fn =3,36 +/- 0,22; Fn+Fag = 3,37 +/-
0,26; p = 0,9767). In the BSS analysis was observed progression of the behaviors of feeding, cleaning and rest without
interruption from them and a slight delay at the satiety point in Fn+Fag group when compared to Sn+Fag.
Conclusions:
The results present in this study suggested that rat subjected to pharmacological manipulation during the development periods
alters mechanisms which trigger satiety behavior in response to increased concentrations of brain serotonin. With our results we
reinforce the importance of the serotonin as a programmer of the brain functions.
Keywords: FLUOXETINE, SACIETY, BEHAVIOURAL SATIETY SEQUENCE, FOOD INTAKE, RATS
Resumo:24-084
ACUTE INTERMITTENT HYPOXIA INCREASES SKELETAL MUSCLE PROTEIN BREAKDOWN IN RATS
Przygodda, F. 1; Manfredi, L. H. 1; Gonalves, D. A. P. 1; Zanon, N. M. 1; Bonagamba, L. G. H. 1;

Machado, B. H. 1; Kettelhut, I. D. C. 2; Navegantes, L. C. C. 1
2
Departamento de Bioqumica e Imunologia - FMRP, USP
1
Departamento de Fisiologia - FMRP, USP
Objectives:
The process of adaptation to hypoxia involves changes in endocrine and metabolic functions. Although it is well known that
carbohydrate metabolism is profoundly altered by hypoxia, little is known about the in vivo effects of hypoxic stress on skeletal
muscle protein metabolism. Thus, the present study was undertaken to examine in rat isolated muscles the possible changes in
proteolysis and protein synthesis that occur after acute intermittent hypoxia (AIH).
Three-week-old rats were exposed to 8 hours of AIH (6% O2 for 40s at 9 min intervals). At the end of the cycle, animals were
sacrificed and the soleus and extensor digitorum longus (EDL) muscles were harvested. Then, rates of protein synthesis and
degradation, the activity of proteolytic systems, protein levels and the profile of Ubiquitin(Ub)-proteasome system (UPS)-related
and autophagy-related genes were examined. In both soleus and EDL muscles, rates of overall proteolysis (nmol Tyrmg wt-12h1) increased after AIH. This rise in EDL was accompanied by a 40% and 48% increased activity of UPS (0,100 0,006 vs 0,140
0,009) and Ca2+-dependent proteolysis (0,056 0,0013 vs 0,107 0,028), respectively. Furthermore, AIH induced a 20%
increase in the levels of Ub-protein conjugates and a 2-fold increase in gene expression of atrogin-1 and MuRF-1, two key Ubprotein ligases involved in muscle atrophy, and LC3, a marker of autophagy. In vitro rates of protein synthesis (nmol Tyrmg wt12h-1) in EDL muscles did not differ between AIH and control rats (0,231 0,019 vs 0,229 0,012).
Conclusions:
The data are in agreement with the notion that hypoxia acts as a catabolic trigger on muscle protein metabolism and show that the
UPS and Ca2+-dependent pathway play an important role in AIH-induced muscle proteolysis and suggest that ubiquitin ligases
and autophagy may participate in the development of muscle wasting during chronic hypoxia.
Keywords: HYPOXIA, PROTEIN METABOLISM, SKELETAL MUSCLE
Financial Support: CAPES and FAPESP (08/06694-6).
Resumo:24-085
PERINATAL PROTEIN RESTRICTION MAY INDUCE OXIDATIVE STRESS IN HEART AND LIVER?
Filho, R. C. S. 1; Nascimento, L. C. P. 1; Silva, A. B. 1; Freitas, C. M. 1; Matos, R. J. B. 1; Leandro, C. G. 1;

Lagranha, C. J. 1; Fernandes, M. P. 1
1
Physical Education Nucleus and Sports Science, CAV/UFPE
2
Nutrition Nucleus, CAV/UFPE
Objectives:
The mechanism of the association between malnourish and risk of disease in adulthood remains controversial. Given that our aim
was evaluate the effect of malnourish (i.e. protein restriction) during critic period of development in the oxidative stress in heart
and liver.
Male Wistar rats were used and divided according to the diet received during the perinatal period. Control group (animals from
mothers that received 17% casein during the gestation and lactation period), and malnourished group (MG: animals from mothers
receiving 8% casein diet during the gestation and lactation period). After this period both groups were given standard animal
house diet (Labina-Purina). Samples from heart and liver were obtained from control and malnourished-rats. At 90 days of age
the animals from both group was sacrificed, and heart and liver were quickly removed and saved in -80oC until biochemical
analysis. We evaluate protein concentration by Bradford method (Anal Biochem. 72:248-254, 1976), levels of oxidative stress
(Malondialdehyde-MDA previously described by Draper & Hadley in Methods Enzymol 186:421-31, 1990) and antioxidant
enzyme (catalase activity-CAT previously described by Aebi et al. in Methods Enzymol 105:121, 1984). We observe that in heart
a non-significant increase in MDA in the malnourished group (MG: 8.15 + 1.09 vs control group: 5.16 + 1.4 nmol/mg protein)
and a significant increase in catalase activity (MG: 330.9 + 24.15 vs control group: 171.4 + 4.3 nmol/min/mg protein;
pnmol/min/mg protein).
Conclusions:
Our data suggest that protein restriction during critic period of development induce significant alteration in oxidative stress
biomarker in heart which could be associated with diseases in adulthood.
Keywords: Heart, Liver, Oxidative Stress, Protein Restriction
Resumo:24-086
APPLICATION OF LAB-ON-A-CHIP TECHNOLOGY FOR PROTEOMICS ANALYSIS OF BLOOD SERUM OF
ADULT WISTAR RATS SUBMITTED TO NEONATAL MALNUTRITION
Raele, R. A. 1; Bezerra, A. A. 2,1; Firmino, G. O. 1; Andrade, M. A. 5,1; Martins, D. B. G. 4,1; Chaves, M. E.

C4,1; de Castro, C. M. B. 3,1; Lima-filho, J. L. 2,1,4
1
Laboratrio de Imunopatologia Keizo Asami, LIKA/UFPE
2
Centro de Cincias Biolgicas UFPE, CCB/UFPE
3
Departamento de Medicina Tropical- UFPE, DMT/UFPE
4
Departamento de Bioqumica - UFPE, DBQ/UFPE
5
Departamento de Fisioterapia- UFPE, DFisio/UFPE
Objectives:
During the intrauterine and neonatal life (critical periods of development), environmental factors - for example, nutrients from the
diet - trigger permanent changes in physiology and metabolism, influencing the development of animals. Physiological changes
can be monitored through the search for protein biomarkers. In this context, proteomic techniques - such as the lab-on-a-chip
electrophoresis system - are important tools for the study of expression of differential proteins. Thus, the aim of this study was to
evaluate, through lab-on-a-chip electrophoresis technology, the one-dimensional proteomic profile (ODPP) of blood serum from
adult rats submitted to neonatal malnutrition (NM).
8 Wistar and male rats were used, obtained newborns from the vivarium of the Department of Nutrition (UFPE), divided into two
groups according to the administered diet durring lactation period (1 to 21 days of life): Nourished Group (NG), consisting of
pups whose mothers were fed with diet containing 17% of casein and Malnourished Group (MG), in which mothers were fed
with diet containing 8% of casein. After the lactation period (22 day), began the nutritional supplementation period in which
pups were fed with a standard vivarium diet (23% protein). Animals were weighed daily during lactation and on alternate days
for nutritional supplementation period. On day 90 of life, were anesthetized with a combination (1:1) of ketamine(116g/mL) and
xylazine (23g/mL) chloridrates, and then, subjected to cardiac puncture for blood collection (4mL). After coagulation and
centrifugation, the serum layer was collected and packed at -20 C. To obtain the ODPP serum, aliquots were diluted (1:10) and
deposited on the chips, using the kits Agilent Protein 80 (for proteins 50 to 80 kDa) and Agilent Protein 230 (for proteins between
14 and 230 kDa), followed manufacturer's instructions. Were then applied to the Agilent Bioanalyzer 2100 appliance and the
results obtained in the form of gel of electropherogram. For data analysis, we calculated the molecular medium weight (MW) of
the proteins (kDa) and the tests used were t-test and Mann-Whitney test (p0.05) to compare the protein concentrations (ng/L)
of both groups. It was also used the TagIdent tool (ExPASy) to search for a possible proteins present in the bands of MW
detected. MG animals showed body weight values lower than NG from the 5th day of life (NG: 12.553.35g and MG:
10.412.03g, p=0.0262), condition that remained until day 90 (p
Conclusions:
The (NM) caused a permanent loss of body weight until day 90 of life, and in the adult animal, caused changes in the ODPP of
the serum samples analyzed. These changes in the proteome profile may be related to changes in the metabolism of MG animals
which potentially are associated with development of metabolic disorders in adults.
Keywords: LAB-ON-A-CHIP Technology, Biomarkers, Neonatal Malnutrition, Proteomic Profile
Resumo:24-087
THE OXIDANT ACTIVITY OF ALVEOLAR MACROPHAGES IN OBESE RATS SUBJECTED TO ENDOTOXEMY
Sales, T. H. A. ; Silva, R. R. ; Silva, K. M. F. ; Santana, V. O. ; Barros, V. A. ; Andrade, M. A. ; de Castro,

C. M. M. B.
Laboratrio de Imunopatologia Keizo Asami - LIKA, UFPE
Objectives:
To observe the effects of obesity related to the oxidant activity of alveolar macrophages in endotoxemic rats.
We used male Wistar rats (n=40), aged 147 days. After weaning, the animals were divided into two groups according to the
dietary regimen. The diet group (DP) was fed with the standard diet (Labina-Purina of Brazil S/A) and the hyperlipidic diet group
(DH) was maintained with a high fat diet consisting of a mixture calorie diet (normal protein and hyperlipidic). The calorie
density to the diet palatable was 4.8kcal/100g (24.5% Lipid) whereas the standard diet, presented 2.7kcal/100g (4% Lipid). The
management of the diets lasted 18 weeks, during which time all animals were weighed on alternate days. Afterwards the groups
were subdivided into endotoxemic (DPE E DHE) and not endotoxemic (DP and HD). To obtain the endotoxemic groups, animals
received intraperitoneal injection of lipopolysaccharide at a dose of 1mg/kg of body weight. After 24 hours, the bronchoalveolar
lavage was collected and from two isolated macrophages, the dosiphication of nitric oxide and superoxide was carried out. The
production of nitric oxide was determined according to the supernatant cells in cultivation. The cultivation plates were placed in
the stove for 24 hours so to be read in the spectrephotometer. The production of superoxide was appreciated in two instances (1h
and 2h) through their ability to reduce in vitro the ferrocitocrome C, after being stimulated in acetate miristate of forbol. In the
comparison of the two groups, the Student T test was used for the parametric data and the Mann-Whytney test for the nonparametric data. The results were expressed as means standard error of the mean. The DH group, in opposition to the DP one,
did not present alteration in the body weight (DP=273,68,9g and DH=268,75,5g). However, the overlipid diet of the animals
increased the quantity of the visceral fat as the DH group was found thrice fatter than the DP one (DH=14,50,8g; DP=4,80,3g).
The nutritive quantity of the DP group (22,03,2g) was more than 32% of the DH group (15,0 1,4g,P=0,001). The DH animals
presented a reduction of the NO production in comparison to the DP ones (DP=9,880,49; DH=7,770,68,P=0,024). When these
animals were subjected to endotoxemia, the DPE group showed a considerable reduction of the production of NO compared to
the DP one(DPE=8,751,24; DP=9,880,49,P=0,034). Amongst the endotoxemic animals, the production of NO in the group
DHE showed an important reduction when it was compared to the DPE group (DHE=6,211,52; DPE=8,751,24,P=0,001). The
production of superoxide was manifestedly reduced (P=0,001) in DH animals in comparison of DP animals after 2 hours of
stimulation of the macrophages with PMA. During the first hour, the DP and the DH group differed in the expulsion of
superoxide (DP=1,220,14; DH =0,670,23,P=0,001). In the second hour, the expulsion of superoxide remained altered when the
two groups were compared (DP=1,420,20), being higher in the DH group (0,800,17,P=0,001). Between the two endotoxemic
groups, the group DHE (0,631,99) presented a reduction in relation to the DPE one (1,310,69,P=0,020) during the second hour.
Conclusions:
The decrease in nitric oxide and superoxide production by alveolar macrophages in response to the development of obesity show
that there is an impairment in the activity of macrophages and consequently a loss in the host response against an infectious
process.
Keywords: overlipid palate diet, cellular immunology, infection, oxidative stress
Financial Support: FACEPE, CNPq, PROPESQ.
Resumo:24-088
THE USE OF A CHILDS FEEDING QUESTIONNAIRE (CFQ) FOR THE STUDY OF PARENTAL ATTITUDES,
BELIEFS AND PRACTICES ABOUT CHILDS FEEDING AND OBESITY PRONENESS
Lorenzato, L. 1; Braga Costa, T. M. 2; Almeida, S. S. 1

1
Departamento de Psicologia - FFCLRP-USP , FFCLRP-USP
2
Curso de Nutrio - Universidade de Ribeiro Preto, UNAERP
Objectives:
To investigate the attitudes, beliefs and practices of the parents about childs feeding and obesity proneness on children who are
served by the Unified Health System (UHS) in the city of Ribeiro Preto - SP.
This project was approved by the Institutional Ethics Committee (CEP-FFCLRP n452/2009). Three hundred parents and
children (150 parents and 150 children of both sexes) were the participants. The children were randomly chosen, with the age
ranging from 2 to 11 years. The instrument used was the Child Feeding Questionnaire (CFQ), that assesses parents beliefs,
attitudes and practices about childs feeding and these findings were compared with the trend towards childhood obesity. The
original version of this questionnaire has 31 questions and it evaluates 07 factors: Perception of Responsibility, Parents Weight
Perception, Childs Weight Perception, Concerning about Childs Weight, Restriction, Pressure on Eating and Eating Monitoring.
The QAC was applied on parents and the anthropometric measurements were recorded by measurement of parents and
childrens body weight and height, according to the instructions recommended by the Ministry of Health. For the data analysis, a
descriptive statistics (percentages and frequencies), the Kendall correlation test, when applicable, was used. The levels of
statistical significance were set at p < 0.05. The assessment of nutritional status used the Body Mass Index (BMI) according to
the classification criteria recommended by the World Health Organization (WHO). The results showed that 17.33% of the
children and 27.33% of the parents were overweight. In relation to the QAC, for the Perception Responsibility Factor, mothers
considered themselves, the responsible ones for feeding their child most of the time. As for Parents Weight Factor, they reported
to have had normal weight from childhood up to the present moment. The Childs Weight Factor Perception, parents also
affirmed that their children always had normal weight from their childhood up to the present moment. On Concerning about
Childs Weight Factor, the mothers are considered to be worried. About the Restriction Factor, mothers agree with the practice to
restrict the ingestion of some type of food to their children. For the Pressure on Eating Factor, mothers agree about the pressure to
regulate the quantity of determined type of food consumed by their children. On Monitoring Factor, mothers reported that, they
always should monitor what their child eats. Moreover, there was a positive correlation between children and parents BMI
variables, as well as childrens BMI and the factors Perceived on Responsibility; Childs Weight; Concern about Child's Weight,
Restriction and Monitoring, and negative correlation between childrens BMI and Pressure on Eating.
Conclusions:
It can be concluded that the CFQ is an useful tool to measure parental attitudes, beliefs and practices about childs feeding and
obesity proneness, and the present results suggest that the parents behavior on their childs feeding are positively associated with
overweight in childhood.
Keywords: childhood obesity, feeding behavior, nutrition
Financial Support: CAPES.
Resumo:24-089
SERUM LEVELS OF MICRONUTRIENTS AND THYROID FUNCTION IN DOWN SYNDROME
Calil, N. O. 1; Sarkis, J. E. D. S. 2; Hortellani, M. A. 2; Chicourel, E. L. 1; Raposo, N. R. B. 1

1
NUPICS/Faculdade de Farmcia, UFJF
2
Centro de Qumica e Meio Ambiente, IPEN
Objectives:
Down syndrome (DS) is the most common genetic alteration at birth, occurring in all races and socioeconomic levels. About 30%
of individuals with DS develop hypothyroidism and this fact may be related to changes in serum levels of micronutrients such as
zinc (Zn), copper (Cu), iron (Fe) and magnesium (Mg).The aim of the study was to determinethe serum levels of these
micronutrients in patients with DS and to show whether there is a connection between these ones and the occurrence of
hypothyroidism in this population.
Cross-sectional study conducted with DS subjects, between 5 and 15 years of age, attended by the public health system of Juiz de
Fora - MG, and their respective well age and sex-matched controls, selected among the students of this city. Whole blood (10
mL) was collected and centrifuged (1258 g/10 min/4C) to separate the serum. This one was diluted with 1% HNO31:4 (w/w) for
Zn, Cu and Fe measurements and 1:40 (w/w) for Mg measurements. Micronutrients were quantified using flame atomic
absorption spectrometry, model Spectr-AAS-220-FSVarian.The instrumental parameters: wavelength of 213.9 nm (Zn), 324.8
nm (Cu), 248.3 nm (Fe) and 285.2 nm (Mg); with a deuterium lamp background correction; air/acetylene flame; analysis were
performed in duplicate with integration time of 4 seconds. The validation of this method was performed by analyzing human
serum reference materials (Seronorm, QMEQAS06S-06 and QMEQAS08S-06) and human serum samples from the proficiency
test organized by the Wadsworth Center New York State Department of Health. The range of recovery for serum samples and
certified reference materials were 85-103%. Descriptive statistical analysis of serum levels of micronutrients and analysis of
variance (ANOVA), followed by Tukey's post hoc test were performed using the software Statistical Package for the Social
Sciences version 14.0. The minimum signicance level was set at p < 0.05. The study was approved by the Ethics Committee of
the Federal University of Juiz de Fora (015/2010). The sample included 27 subjects with DS (age 9.83.6 years) and 35 age- and
gender-matched healthy controls (9.02.8years). There were no statistical significant differences between groups in serum levels
of Zn (DS: 67.8 11.8 g/dL; control: 75.2 18.1 g/dL), Cu (DS: 110.9 20.7 g/dL; control: 100.4 17.2 g/dL), Fe (DS:
114.0 35.4 g/dL; control: 109.2 47.7 g/dL) and Mg (DS: 1758.9 102.9 g/dL; control: 1720.5 176.9 g/dL). However,
significant differences in ratios Zn/Cu (p=0.002), Zn/Fe (p=0.012) and Zn/Mg (p=0.024) were found between the groups.DS
subjects were classified according to thyroid status as euthyroidism (n=15) and hypothyroidism (n=14). There were no statistical
significant differences between these groups in serum levels of Zn, Cu, Fe and Mg, as well as in the proportion of these
micronutrients.
Conclusions:
Altered serum levels of micronutrients were observed in Down syndrome, but they did not relate to thyroid status in this
population.
Keywords: Down syndrome , Micronutrients, Thyroid disease
Resumo:24-090
THE ANTIOXIDANT QUERCETIN REDUCES DNA DAMAGE IN NONALCOHOLIC STEATOHEPATITIS IN
MICE
Marcolin, . 2,3; Forgiarini, L. F. 3; Tieppo, J. 3,4; Rodrigues, G. 1,3; Picada, J. 6; Freitas, L. A. R. D. 5;

Tuon, M. J. 4; Gonzlez-gallego, J. 4; Marroni, N. P. 1,2,6,3
1
Programa de Ps Graduao em Medicina, PPGCM/UFRGS
2
Programa de Ps Graduao em Fisiologia, PPG Fisiologia/UFRGS
3
Laboratrio de Fisiologia Experimental e Gastroenterologia, LFEG/HCPA
4
CIBERehd, CIBERehd/UNILEON
5
FIOCRUZ, FIOCRUZ - CPqGM
6
Programa de Ps Graduao em Gentica e Toxicologia Aplicada, PPGGTA/ULBRA
Objectives:
Nonalcoholic steatohepatitis (NASH) is a metabolic condition characterized by lipid accumulation in hepatocytes, inflammatory
infiltration and fibrosis and with increased frequency in individuals with obesity, overweight and metabolic syndrome. You can
verify that the DNA damage may be one of the factors involved in this disease. The aims was verify the biochemical levels,
histopathology, lipid peroxidation and DNA damage in mice with NASH induced by a diet deficient in methionine and choline,
treated with the flavonoid quercetin (Q).
METHODS: We used C57BL/6 males, 8 weeks, divided into four experimental groups (n=12) CO+V (control + vehicle
carboxymethylcellulose), CO+Q (Q 50 mg/kg), NASH+V, NASH+Q. Was administered intragastrically 250 ul of Q for 4 weeks.
Were analyzed in liver tissue lipoperoxidation by TBARS (Thiobarbituric Acid Reactive Substances), liver function tests - Aspart
Aminotransferase (AST), Alanina aminotransferase (ALT) and alkaline phosphatase (ALP) and histologic hematoxylin-eosin,
Comet Assay to investigate damage to DNA. Analysis of TNF-alpha; and COX-2 mRNA expression was performed in liver
tissue by quantitative real time RT-PCR. The study was conducted with the approval of the Ethics and Research of Hospital de
Clnicas de Porto Alegre. Data are presented as meanSEM and statistically analyzed by ANOVA followed by Student
Newmann-Keuls with significance of 5%. RESULTS: Lipid peroxidation, blood AST, ALT and ALP activities and mRNA levels
of pro-inflammatory cytokines were significantly higher in untreated NASH+V rats compared to CO+V rats. The flavonoid
quercetin significantly lowered lipid peroxidation (7,671,84 vs 6,80,8(mol/mg de protein), serum ALT (534,7860,83 vs
36817,38 U/l), AST (481,2547,68 vs 352,1739,24 U/l) and ALP (1082,661 vs 96,293,27 U/l), activities and consistently
caused a reduction of mRNA levels of TNF-alfa (737,6338,1 vs 418,5719,1) and COX-2 (720,977,1 vs 355,140,8), in
NASH animals compared with treated NASH+Q rats. It is observed histological improvement in NASH+Q group compared to
the NASH+V. And, there were reductions in both the damage index as the frequency of DNA damage in the NASH+Q group
compared with NASH+V group (189,118,2 vs 215,811,9%).
Conclusions:
The administration of Q 50mg/kg shows improvement in the liver of NASH due to a decrease in lipid peroxidation, indices of
liver injury and in pathological examinations. This effect can be explained by the high antioxidant ability of quercetin and your
action in the degree oxidative stress in this patology.
Keywords: QUERCETIN, NONALCOHOLIC STEATOHEPATITIS, ANTIOXIDANT
Financial Support: CAPES-REUNI; HCPA; CIBERehd
Resumo:24-091
CONSUMPTION OF SOFT DRINKS REDUCES PROTEIC AND CALORIC CONSUMPTION FROM SOLID FOODS
IN RATS WISTAR WITH OBESITY INDUCED BY CAFETERIA DIET
Mucellini, A. B. ; Goularte, J. F. ; Sanvitto, G. L.

Graduate Program in Biological Sciences: Physiology, UFRGS
Objectives:
Recent investigations indicate that the soft drink contributes most total daily energy intake. Besides being associated with weight
gain and energy intake, its consumption also appears to be associated with changes in food choices, since it was positively
associated with intake of processed foods and negatively with intake of fruits, vegetables, grains, fish, seafood and eggs. Some
authors suggest that the obesity epidemic, observed mainly in the United States, may have been the result of increased
consumption of soda. The cafeteria diet composed of palatable foods and calorie-rich carbohydrates and lipids, including soda, is
an animal model that reproduces the approximate feeding pattern observed in many countries and is associated with the obesity
epidemic. In a previous study, we compared the pattern of nutritional and caloric intake of rats with access to two diets: standard
diet (SD) (only food and water) and Cafeteria Diet (CD) (food, water, food palatable and soda).
Female Wistar rats (n = 24) were fed from weaning with CD or SD for 26 weeks. The foods offered on CD were: chow, Salami,
Bread Seven Boys, Snack Yokitos, Deliket Jelly Bean, Coca-Cola, Smoked Sousage, Chocolate Cake, Biscuit Maizena,
Marshmallow, Ham, Snack Fritello, Wafer Biscuit Chocolate and Gumdrop Gomets. The assessment of food intake was
performed by daily weighing of each food. From the data of weight and volume of each food ingested and the nutritional
information listed on product labels, it was possible to quantify the total energy intake, carbohydrate intake, protein intake, intake
of fat and sodium intake. At the end of week 26, the intake of standard food and water from the CD group was lower compared to
the SD group (3.2g 0.2 vs.14.5g 0.2 e 6mL vs. 22.2mL, p<0.05).
Conclusions:
The reduction in caloric intake of solid food observed in the CD group compared to the SD group, indicates that the effects of
intake of soda on food intake may be more harmful than just produce increased intake of energy, such as the decrease of protein
intake, due to low content of this nutrient on CD.
Keywords: Cafeteria diet, Obesity, Soft drinks, Proteic consumption, Caloric consumption
Financial Support: CNPq and Graduate Program in Biological Sciences: Physiology/UFRGS
Resumo:24-092
MATERNAL HIGH-FAT AND/OR HIGH-SALT DIET AND GLUCOSE METABOLISM IN MICE OFFSPRING
Bringhenti, I. ; Moraes-teixeira, J. A. ; Leiroz, R. F. ; Cunha, M. R. ; Mandarim-de-lacerda, C. A. ; guila,

M. B.
Departamento de Anatomia, UERJ
Objectives:
Maternal nutrition is a strong factor affecting progeny. This study aimed to assess how the maternal high-fat and/or high-salt diets
alter the biometry and glucose metabolism.

Six weeks-old C57BL/6 female mice were divided into four groups according to the amount and type of lipid and salt in the diet:
SC (Standard Chow), HS (High-salt), HF (High-Fat), HF-HS (High-Fat and High-salt). The diets were administered from 8-wks
before mating and throughout pregnancy and lactation periods. Male offspring was assessed at birth and at 10 days old. No
difference was observed on the body mass (BM), at birth among the groups. However, at 10 days old pups from HF group were
heavier than SC group (+20%, P
Conclusions:
These preliminary results demonstrated that diets administered before and during pregnancy and lactation periods have effects on
glucose metabolism and body biometry, possibly with adverse consequences later in life.
Keywords: maternal nutrition, glucose metabolism, biometry, mice offspring
Financial Support: CNPq, CAPES
Resumo:24-093
EFFECTS OF THE INCREASED BRAIN AMMONIA IN THE METABOLISM.
Serafim, A. C. A. 1; Almeida, M. M. A. D. 1; Cameron, L. C. 2; Lima, N. R. V. 1

1
Educao Fsica, Fisioterapia e Terapia Ocupacional - UFMG, UFMG
2
Laboratrio de Protna e Bioqumica - Uni-Rio, Uni-Rio
Objectives:
Hyperammonemia is a condition common to pathological liver disorders and intense or exhausting exercise. Hyperammonemia is
linked to impairment of normal brain function and the onset of the neurological condition. High levels of brain ammonia can lead
to alterations in cerebral energy metabolism and neurotransmission, which may in turn impact on the functioning of important
signaling pathways within the neurons. Therefore, the aim of this study was to evaluate the effects of central ammonia increase
on the regulation of metabolic responses in rats.
Male Wistar rats (300 10 g; n=6) were housed in individual cages under 1410 h light/dark cycles and had free access to water
and rat chow (CETEA:236/2010). Under Ketamine (80 mg/kg body weight, i.p.) and Xilazine (15 mg/kg kg body weight, i.p.)
anesthesia, a silastic catheter was inserted through the jugular vein into the right atrium for repeated blood sampling. After this
procedure, the rats were fixed to a stereotaxic apparatus, and a guide cannula (22 G) was then implanted into the right lateral
cerebral ventricle. All animals were allowed to recover for at least 1 week before being submitted to the experiments. On the day
of the experiment, an injection needle (30 G) 0.3 mm longer than the guide cannula and connected to a Hamilton syringe was
introduced, via the cannula, into the lateral cerebral ventricle. Either 2.0 L of 0.15 M NaCl (SAL), 2.0 L of 50 M Ammonia
(AM 50 M), 2.0 L of 400 M Ammonia (AM 400 M) or 2.0 L of 750 M Ammonia (AM 750 M) were randomly injected
into the lateral ventricle. Blood was withdrawn from the atrial catheters 10 min before intracerebroventricular (i.c.v.; BASAL)
administration of saline or ammonia and 6 and 16 minutes after the administration. Concentrations of blood glucose, lactate, uric
acid, urea and total protein were measured. The mean values from each experiment trial were analyzed by two-way ANOVA with
repeated measures, and the significance of the differences between groups means was determined by the post hoc Newman-Keuls
test. Throughout the analyses, P
Conclusions:
High levels of brain ammonia lead to alterations on the energy metabolism. The disruption of energy metabolism by central
ammonia involves the decrease the mean blood glucose and lactate to buffer and decrease the elevated ammonia concentrations.
Keywords: Brain, Metabolism, Ammonia
Financial Support: FAPEMIG, CAPEs, PRPG/UFMG
Resumo:24-094
FOOD INTAKE AND ANTHROPOMETRIC PARAMETERS OF RATS FED WITH A DIET BASED ON
HYDROGENATED VEGETABLE FAT DURING GESTATION AND LACTATION PERIODS.
Ladislau, H. F. L. ; Melo, B. M. O. ; Santos, T. D. ; Pereira-da-silva, M. S. ; Borba, J. M. C. ; Rocha-demelo, A. P.

Depto. de Nutrio - Lab. de Fisiologia da Nutrio -LAFINNT, UFPE
Objectives:
Aim: Investigate if the substitution of soybean oil in the diet for hydrogenated vegetable fat alters food intake and murinometric
parameters of Wistar rats.
Methods and Results: During pregnancy and lactation, Wistar rats were fed an isocaloric/normolipidic diets containing 7%
soybean oil (control, C) or hydrogenated vegetable fat 7% (experimental; E). After weaning (21 days old), pups received the
same diet of their respective mothers until adulthood, corresponding to the C (n = 24) and E (n = 38) group. Food intake (g) and
body weight (g) of the dams (C: n = 4 and E: n = 5) were monitored weekly during pregnancy and lactation. Pups were weighted
at the 1st, 7th, 14 th and 21st days of life and after weaning were weighted at 30, 60 and 90 days old. During nine weeks after
weaning, food intake of the male pups (C=4 e E= 6) was checked weekly. At 90 days, rats were anesthetized with Chloralose +
Urethane at dose of 1ml/100g for verification of the following murinometric parameters: head-anus length, chest and abdominal
circumference. The body mass index (BMI) of animals was also calculated. During pregnancy, food intake (g) of the E group was
significantly higher at the 2nd week (E = 158.57 16.21 and C = 113.59 4.84; p < 0.05; mean SEM), whereas during lactation
there was no difference. The body weights (g) of female rats during pregnancy and lactation did not show any difference. At the
day 1 after birth, pups of the E group had lower body weights (g) when compared to the C (E=6.61 0.70 and C=7.02 0.51; p <
0.05; mean SEM). However, at 90 days of life the E group showed higher body weights (g) than the C group (E=296.30
41.27 and C=275.93 49.13; median-interquartile; p < 0.05). At the 1st (E = 88.14 8.15 and C = 45.16 9.08; p < 0.05; mean
SEM), 6th (E = 142.68 3.31 and C = 125.51 5.07; p < 0.05 ; mean SEM) and 7 th (E = 195.35 23.92 and 119.07 8.20; p <
0.05; mean SEM) week after weaning, food intake (g) of pups in the E group was higher than in the C group. There were no
significant differences of the murinometric parameters as well as the BMI between groups.
Conclusions:
Conclusion: The chronic intake of a normolipidic diet based on the hydrogenated vegetable fat during gestation and lactation did
not affect body weight and maternal consumption. However, food consumption and body weight of the pups were influenced by
this diet without change both murinometric and BMI parameters.
Keywords: Food Intake , Hydrogenated Vegetable Fat, Murinometric Parameters, Rats
Resumo:24-095
CHANGES IN LIVER MITOCHONDRIAL METABOLISM IN MICE SUPPLEMENTED WITH CONJUGATED
LINOLEIC ACID
Pereira, A. F. ; S, L. L. ; Prado, I. M. R. ; Alberici, L. C.

Faculdade de Cincias Farmacuticas de Ribeiro Preto, FCFRP/USP
Objectives:
Several studies have shown that daily intake of conjugated linoleic acid (CLA) reduces body fat accumulation, increases body
metabolism and prevents obesity in animal models(Int J Obes Relat Metab Disord. 28:941, 2004). Dietary supplementation with
CLA reduces lipid accumulation in adipose tissue and increases the supply of free fatty acid to other tissues such as liver(Lipids.
32:853, 1997). The aim of this study was to determine whether the increased supply of lipids to the liver alters the parameters of
liver mitochondrial metabolism. The hypothesis is that supplementation with CLA induces mechanisms of mitochondrial
uncoupling.
Four groups of 10 male C57BL6 mice, aged 5 weeks, had access to standard laboratory rodent chow containing 4% lipids (CR1;
Nuvital, Colombo, Paran, Brazil) and water ad libitum and were housed at 22C 2C on a 12-hour light-dark cycle. Each
group was divided into two subgroups of 5 animals supplemented with 0.05 mL linoleic acid (control) or CLA (50% of each
isomer cis-9 trans-11 and trans-10,cis-12) by gavage, 3 times/week for 2 months. The dose of CLA administrated corresponds for
1% of total diet. The mice had their body weight gain monitored once a week. After this period the mice were sacrificed in a CO2
chamber and perigonadal adipose tissue and liver were extracted and weighed. A slice of liver right lobe was used for histology
analysis (hematoxilina/eosina) and whole liver for mitochondrial isolation (differential centrifugation, Biochem J. 212:279,
1983). Mitochondrial oxygen consumption was monitored using a Clark type electrode (Hansatech Instruments Ldt, OXIGRAPH
software V1.10, England) in a thermostatically controlled chamber equipped with magnetic stirrer. Statistical analysis was
performed by Student's t test for two-mean comparison or one-way ANOVA, for multiple comparisons using the GraphPad Prism
5. Supplementation with CLA did not change body weight gain within 60 days (P = 0.3). However, CLA supplementation
increased 12% liver weight (P = 0.0002) and reduced 42% the weight of perigonadal adipose tissue (P = 0.0002). No histological
alterations on hepatic lipids were found in liver from CLA-supplemented mice. Liver mitochondria isolated from CLAsupplemented mice presented 16% (P = 0.033) reduction on respiratory control ratio (RCR, State III/State IV) due to elevation
State IV respiration in the presence of 0.2 M linoleic acid compared to liver mitochondria isolated from control mice. These
results suggest the induction/activation of mitochondrial uncoupling system, such as uncoupling proteins.
Conclusions:
Dietary supplementation with CLA do not reduce body weight but decreases adiposity. In liver, this supplementation increases its
weight, not related to lipid accumulation, and induces mitochondrial uncoupling mechanisms to dissipate the excess of energy
intake.
Keywords: Adiposity, CLA, Metabolism, Mitochondria, Uncoupling
Financial Support: PIBIC/CNPq
Resumo:24-096
ORGANOTIN AND METABOLIC RATES: PRELIMINARY STUDIES
Lopes, P. F. I. 1; Filho, V. S. D. 1; Podratz, P. L. 1; Barros, V. P. O. 1; Tamie, S. M. 2; Costa, M. B. 1;

Takiya, C. M. 2; Graceli, J. B. 1
1
Universidade Federal do Esprito Santo, UFES
2
ICMorfolgicas, Universidade Federal do Rio de Janeiro, ICB, UFRJ
Objectives:
Organotin compounds (OT), such as tributiltin (TBT), have been widely used as biocides, agriculture fungicides, wood
preservatives, and disinfecting agents in circulating industrial cooling waters, as well as antifouling paints for marine vessels.
There are many reports of the biological effects of OT, which vary in their toxic effects on eukaryotes. These compounds are
potent endocrine disrupters in gastropods,and through the inhibition of aromatase enzyme can increase testosterone levels, change
reproductive parameters, and induce mollusks sterile by the development of an endocrine syndrome called imposex. These
chemical compounds are also suspected to cause endocrine-disrupting effects in mammals, due in part the possibility of
transferring through marine food chains. Although reproductive function is mainly regulated by sexual hormones. It is likely that
female gender steroids are correlated with changes in metabolic parameters, for example in normal function of white adipose
tissue. To evaluate the influence of OT on lipid and metabolic profile correlated with changes on female rat steroids hormones.
Wistar fertile female rats were utilized (200-250g, aged 3 months, Rattus norgegicus). Animals were divided in two groups: 1)
Control (CON, n=8, treated daily with vehicle for 14 days by gavage); 2) Treated with TBT (TBT, n=8, 100ng/kg, treated daily
with TBT for 14 days by gavage). After treatment, the animals were sacrificed, their blood was collected, glucose levels, lipid
profile (HDL, LDL, triglycerides, total cholesterol), the perirenal, mesenteric, retroperitoneal and parametrial weight fats were
measured. Also, the white adipose tissue was analyzed by histological sections stained with hematoxylin and eosin. Results are
presented as meanSEM. One-way analysis of variance (ANOVA) followed by unpaired Student t-test). Differences were
assumed to be significant when p
Conclusions:
These results suggest that serum alterations in female sexual steroids, induced by OT, affect metabolic parameters in female rats.
Keywords: female sexual hormones, metabolism, Organotin compounds
Financial Support: UFES , FAPES (n 45446121/09)
Resumo:24-097
LIVER OXIDATIVE PROFILE OF RATS SUBMITTED TO AN HIPERLIPIDIC DIET AND TREATED WITH THE
AQUEOUS EXTRACT OF ACHYROCLINE SATUREIOIDES
Espia, D. C. 2; Pinheiro, K. V. 1; Zanini, D. 1; Carvalho, F. B. 1; Schlemmer, J. B. 1; Kohl, C. O. 2;

Zimmermann, C. E. P. 1; Leal, D. B. R. 1; Baiotto, C. R. 2; Jaques, J. A. S. 1
1
Programa de Ps Graduao em Bioqumica Toxicolgica, UFSM
2
Curso de Cincias Biolgicas, UNICRUZ
Objectives:
To investigate the effect of the aqueous extract of Achyrocline satureioides on the liver oxidative profile of rats submitted to an
hiperlipidic diet.
Twenty four adult male Wistar (200-250g) rats were used in this study. The animals were randomly divided into 4 groups (6 rats
in each group): control (C); A. satureioides 10% (A10), treated with a 10% aqueous extract; Hyperlipidic control (H), treated with
the hyperlipidic ration; and Hyperlipidic/ A. satureioides 10% (HA10), treated with the hyperlipidic ration and the 10% aqueous
extract. The animals from the H and HA10 group received the hyperlipidic ration ad libitum immediately after the weaning (21
days old). The animals from the groups C and A10 received the standard ration equals to 10% of their body weight to avoid the
excessive weight gain. The animals from the groups A10 and HA10 received the aqueous extract of A. satureioides. For the
preparation of the hyperlipidic ration it was grinded 650g of standard ration for rodents, 500 mL of distilled water, animal grease
(200g), soy oil (100 ml) and starch (50g of wheat powder). Latter the ration pellets were prepared and let to dry at 50oC
overnight. The aqueous extract of A. satureioides was prepared as a decoction (10 mg/mL) and was administered orally according
with the animal weight (1 ml/kg of body weight). Six weeks after the hyperlipidic diet started, the aqueous extract administration
began and lasted 5 weeks. At the end of the experiment, the rats, at 12 hours of fasting were anesthetized with isoflurane and the
liver removed to perform the oxidative stress assays. The liver was homogenated (1:10, w:v) with Tris HCl 50 mM pH 7.5,
centrifuged, and in the supernatant it was measured the amount of thiobarbituric acid reactive substances (TBARS) and carbonyl
protein. In the same sample it was measured the enzymatic activity of superoxide dismutase (SOD), catalase (CAT) and Na+,K+ATPase. The data presented as mean + Standard Error of the Mean (SEM) was analyzed with one and two-way ANOVA
followed by the Post Hoc test Student Newman Keuls and considered significant when pA. satureioides was able to prevent these
effects. The CAT activity showed to be decreased in both H (30 %) and HA10 (28 %) groups.
Conclusions:
The data obtained from this study suggest an increased protein oxidation in the rats treated with the hyperlipidic diet, as
demonstrated by the carbonyl content. Another fact which remits to an increased protein oxidation is the decreased activity of Napump in the H group. The decreased activity of the antioxidant enzyme CAT would contribute to the elevated protein damage
observed in the H group. Taken together, these results point toward the antioxidant effects of the aqueous extract of A.
satureioides.
Keywords: Achyrocline satureioides, Hiperlipidic diet , Lipid Profile , Na+,K+-ATPase, Oxidative stress
Financial Support: CNPq, FAPERGS and CAPES
Resumo:24-098
METABOLIC PROFILE OF OVARIECTOMIZED RATS SUBMITTED HIGH-FAT DIETS WITH DIFFERENT
SODIUM LEVELS.
Santos Filho, L. E. ; Santos, R. L. ; Santos, G. P. L. ; Dantas, A. C. S. ; Carvalho, G. O. ; Coimbra, C. C. ;

Soares, T. J. ; Magalhes, A. C. M.
UNIVERSIDADE FEDERAL DA BAHIA, UFBA
Objectives:
Estrogen levels reduction in the post-menopausal period has been associated with metabolic changes that may interfere in the
regulation of body weight, blood pressure, adipose tissue and insulin resistance. High salt consumption contributes to the
pathogenesis of hypertension, but its association with the development of insulin resistance, changes in the plasma lipids
concentrations and in the adipose tissue deposition pattern in the post-menopausal period is still not fully elucidated. In this
context, this study aimed to evaluate the metabolic profile of ovariectomized female rats submitted to high-fat diets with different
sodium levels.
Twenty ovariectomized(O)female Wistar rats were submitted to standard diet (SD), high-fat diet (HF) or high-fat diet with
sodium content of 8% (HFS) for 20 weeks. The groups consisted of ovariectomized female rats submitted to standard diet (OSD,
n = 5), or high-fat diet (OHF, n = 5) or high-fat diet with sodium content of 8% (OHFS, n = 10). The animals were kept in a room
with light (light-dark cycle of 12h) and temperature ( 23C) control and free access to both water and food. Body weight
measures and blood pressure determinations were weekly evaluated. The food, caloric and sodium intake were monthly evaluated
through metabolic assays. The plasma biochemical profile was analyzed through serum glucose, total cholesterol and
triglycerides levels. For evaluation of insulin resistance was held the glucose tolerance test (1g&fraslkg) and plasma insulin by
radioimmunoassay. After animals sacrifice, the visceral adipose tissue (mesenteric, parametrial and retroperitoneal) was collected
weighed and was adjusted for body weight. The results were expressed as mean mean standard error. Statistical differences
were determined using one-way ANOVA followed by StudentNewmanKeuls post hoc (p
Conclusions:
High-fat diet and high-fat high-sodium diet determined increases in the visceral adipose tissue weight and decrease in the glucose
tolerance in ovariectomized female rats. Moreover, high-fat high-sodium diet promoted systolic hypertension despite having
determined decrease in the caloric intake and lower weight gain in relation to high-fat diet.
Keywords: Estrogen, High-fat diet, Metabolic profile, Ovariectomy, Sodium
Financial Support: Conselho Nacional de Pesquisa e Desenvolvimento (CNPq)
Resumo:24-099
FRAIL ELDERLY AND NUTRITIONAL RESTRICTION
Moreno, D. F. 1; Ferreira, A. S. 2; Raposo, N. R. B. 1,2; Barbosa, E. M. S. 3; Chicourel, E. L. 1

1
NUPICS, Faculty of Pharmacy, UFJF
2
Depto de Psiquiatria, Faculdade de Medicina, IPq-HCFMUSP
3
Centro Mais Vida, SES/MG
Objectives:
Aging process is a worldwide reality and a challenge in developing countries including Brazil due to the lack of resource and the
usually inefficient planning. Frail elderly needs a specific and rich food, and many times this nutrition requires restriction due to
diseases, specially hypertension and diabetes, the chronic and most prevalent ones. The aim of this study was to characterize a
frail elderly sample and to check if the nutrition restriction is presented in the cited diseases.
One hundred sixty-one (161) frail elderly were evaluated by questionnaires developed by the researchers and they were applied in
their houses during a research day. All participants agreed to take part and signed the Free Informed Consent. Among the
questions, socio-demographic characteristics, food restriction and auto-related absence or presence of the most common elderly
diseases were answered. Data were expressed as frequencies and mean with standard deviation. To verify association, qui-square
test was performed. If an association was verified, odds ratio with the confidence interval was calculated. The significance was
set at 0.05. All the analyses were performed with thes Statistical Package for Social Sciences for Windows, version 13.0. The
predominance in the sample was female (n= 94) (58%), married (n = 96) (60%), with 71.2 9.7 years old and until four years of
study (n= 130) (81%), which gain a minimum wage (n= 81) (51%) and had an insurance health (n= 83) (52%). Seventy-eight frail
elderly (48%) had a food restriction. We found that sodium restriction was predominant in hypertensive frail elderly (OR= 7.47,
CI95% = 1.70 32.79) and carbohydrate restriction was predominant on diabetes people (OR= 33.06, CI95% = 10.13 107.84).
Conclusions:
We highlighted the importance of an appropriate diet taken by frail elderly, which can also act on the prevention of hypertension
and diabetes.
Keywords: diabetes, frail elderly, hypertension, nutritional restriction
Resumo:24-100
EFFECT OF THE CAFETERIA DIET IN THE METABOLISM OF WISTAR RATS.
Guilherme, C. ; Louzada, S. M. ; Jacobs, S. ; Krolow, R. . ; Sanvitto, G. L.

Departamento de Cincias Fisilogicas , UFRGS
Objectives:
Aim: There are many experimental models that induce insulin resistence and obesity. The model that most closely resembles
human obesity is exogenous obesity, in which the animal receives a higher caloric intake through a surcharge of carbohydrates
and/or fat. This diet is called fast food diet or cafeteria diet. The cafeteria diet produces an increase in total corporal wheight plus
a significant increase in the amount of visceral fat and induces resistence to insulin. This study evaluates the effect of the diet in
the change of corporal weight and in biochemical parameters and whether the effects differ based on gender.
Methods: Wistar rats newly weaned (n=48), males and females, were used. There were 4 animals per box, light cycle was 12h
light/12h dark and controlled room temperature (22 2C) was maintained. The experiment lasted six months. Animals were
divided in Control Group, which had access ad libitum to water and standard chow, and Cafeteria Group, which had access daily
and ad libitum to soda and water and to highly palatable and hypercaloric food, rich in carbohydrates e lipids, offered in alternate
days. Diet A: standard chow, salami, bread, snacks and candy. Diet B: standard chow, sausage, chocolate cake, biscuit and
marshmallow. Diet C: standard chow, ham, snacks, chocolate waffer and gumdrop. Animals were wheighed weekly and the final
average wheights were compared using the t-test. At the end of the experiment each animal was sacrificed in the guillotine, the
visceral fat was collected and wheighed for comparison between the groups using the t-test. Values are presented as mean SEM
and the significance level was p
Conclusions:
Conclusion: The intake of a hypercaloric and highly palatable diet induces increase in corporal weight and in visceral fat weight,
hypertriglyceridemia and decrease in cholesterol concentration, which suggests that diet composition alters metabolism. We did
not observe the influence of gender in the parameters evaluated.
Keywords: obesity, cafeteria diet, visceral fat, biochemical parameters
Financial Support: CNPQ and Physiology Graduate Program UFRGS.
Resumo:24-101
DISTINCT EFFECTS OF CREATINE SUPPLEMENTATION ON SKELETAL MUSCLE AND CARDIOMYOCYTE
OF RATS TREATED WITH DEXAMETHASONE
Guimares F de S1; Moraes, W. M. M. D. 1; Nicastro, H. 1; Souza, P. R. M. D. 3; Junior, A. H. L. 1; Brum,

P. C. 1; Medeiros, A. 2
1
Escola de Educao Fsica e Esportes, EEFE- USP
2
3
Faculdade de Medicina, FMUSP
Objectives:
To evaluate the effects of creatine supplementation on the remodeling of skeletal muscle, glucose homeostasis and cardiac
morphology in rats treated with high doses of dexamethasone.
Wistar male rats (400-415g) were randomized into four groups: Dexamethasone (DEX; n=6), pair feed control (CON-PF; n=6),
Dexamethasone + CR (DEX-CR; n=6), and pair feed CR (CR-PF; n=6). The group DEX and DEX-CR were treated with
5mg/kg/day of dexamethasone, which were administered in drinking water. DEX-CR and CR-PF also received 5g/kg/day of CR
supplementation in drinking water. Both were treated for 7 days. On plantaris muscle were evaluated muscle weight, total protein
and membrane expression of GLUT-4 at baseline state, and the expression of phospo-AKTSer237 by western blot. In the heart
were assessed heart weight total and chamber`s weight, and were measured the diameter of cardiomyocytes and cardiac collagen
fraction. The weight of the plantaris muscle was significantly lower in DEX compared to CON-PF group (DEX 274 27 vs.
CON-PF 406 27 mg; p < 0.05). CR supplementation did not attenuate the atrophy observed in these tissues (DEX-CR 264 24
vs. DEX 274 27 mg). Fasting glucose was significantly higher in DEX when compared with CON-PF group (DEX 7.8 0.6 vs.
CON-PF 5.2 0.5 mmol/l; p < 0.05). Hyperglycemia has been exacerbated with the addition of CR in DEX- CR compared to
other groups (DEX-CR 15.1 2.4 mmol/l; p < 0.05). The total protein expression of GLUT-4 was similar between groups (p >
0.05). However, the DEX group had lower membrane expression of GLUT-4 compared to CON-PF (p < 0.05) and CR
supplementation aggravated this response, as observed in DEX-CR compared to the others groups (p < 0.05). The dexamethasone
treatment decreases significantly phospho-AKTSer273 expression compared with CON-PF group (p < 0.05), and CR
supplementation aggravated this response (p < 0.001). In the heart, CR supplementation attenuated the increase in left ventricular
mass (DEX-CR 1.15 0.35 mg vs. DEX 1.7 0.12 mg vs. CON-PF 1.71 0.8 mg; p < 0.05), and cardiac collagen volume
fraction (DEX-CR 12.9 1.4 vs. DEX 15.8 0.6 vs. CON-PF 11.6 1.0 %; p < 0.05) induced by dexamethasone treatment.
Conclusions:
CR supplementation did not prevent skeletal muscle atrophy and increased the diabetogenic effects. In contrast, the effects on
cardiac tissue were positives, because attenuates morphological changes induced by dexamethasone.
Keywords: Glicorticoid, heart hypertrophy, creatine supplementation, cardiovascular desords
Resumo:24-102
CDC2-LIKE KINASE 2 (CLK2) IS A KEY MODULATOR OF LEPTIN AND INSULIN SIGNALING/ACTION IN THE
HYPOTHALAMUS.
Quaresma, P. G. F. 1; Santos, A. C. 1; Weissmann, L. 1; Caricilli, A. M. 1; Morari, J. 1; Velloso, L. A. 1;

Mendes, M. C. 1; Saad, M. J. A. 1; Prada, P. O. 1,2
1
Faculdade de Cincias Mdicas, UNICAMP
2
Faculdade de Cincias Aplicadas, UNICAMP
Objectives:
The aim of the present study was to investigate in vivo firstly the effect of insulin and leptin on Clk2 regulation and on energy
homeostasis and secondly the effect of nutritional status and obesity on the modulation of this kinase in the hypothalamus.
By western blot and immunofluorescence we detected Clk2 expression in all hypothalamic nuclei. Fasting decreased and
refeeding increased Clk2 pTh/Ser in the hypothalamus from control mice. Intracerebroventricular (ICV) injection of insulin or
leptin increases hypothalamic Clk2 pTh/Ser in a time and dose dependent manner. ICV PI3K (LY294002) or Akt (Akt VIII)
inhibitors blocked insulin-induced Clk2 pTh/Ser suggesting that the PI3K/Akt pathway mediates the effects of insulin on Clk2
pTh/Ser. Furthermore, the inhibition of Clk2 for 5 days with ICV oligonucleotide antisense (ASO) or with TG003
(pharmacological inhibitor) increased body weight and adiposity due to increased food intake. The inhibition of Clk2 by ICV
ASO or TG003 decreased by 30-40% the anorexigenic effects of leptin and insulin and was associated with lower POMC, CART
and CRH levels in the hypothalamus. In addition, in db/db mice and in diet induced obesity hypothalamic insulin-induced Clk2
pTh/Ser in the hypothalamus was downregulated.
Conclusions:
In summary, our data provide evidence, in vivo that hypothalamic Clk2 pTh/Ser is regulated by nutritional status and by insulin
and leptin. This regulation is blunted in obese mice. Moreover these findings suggest that hypothalamic Clk2 is required for the
appropriate expression of specific hypothalamic neuropeptides to modulate food intake. Thus, the potential for hypothalamic
Clk2 as a site for new therapeutical approach of obesity and energy balance deserves further exploration
Keywords: Clk2, hypothalamus, insulin, leptin
Financial Support: FAEPEX, FAPESP
Resumo:24-103
TUB REGULATION EFFECTS OF CALORIC RESTRICTION AND HIGH-FAT DIET IN HYPOTHALAMUS AND
ADIPOSE TISSUE
Santos, A. C. 1; Quaresma, P. G. F. 1; Weissman, L. 1; Caricilli, A. M. 1; Mendes, M. C. 1; Cesquini, M. 1;

Prada, P. O. 1,1
1
Faculdade de Cincias Mdicas, Unicamp
2
Faculdade de Cincias Aplicadas, Unicamp
Objectives:
The first aim of this study was to investigate whether the caloric restriction (CR) or high-fat diet (HFD) modulate the regulation
by insulin of tub in mice hypothalamus. Recently it was shown that tub is regulated by insulin, in vitro, in adipose tissue cells.
Therefore, the second aim of this study was to investigate the tub regulation by insulin, in vivo, using adipose tissue of control
and obese mice.
The CR was achieved by reduction of 30% of chow and high fat diet included 55% of calories from saturated fat. Both diets were
administered for 8 weeks. The extraction of the hypothalamus was performed after intracerebroventricular injection of insulin,
leptin or saline and the tyrosine phosphorylation of tub (p-tyr tub) were analyzed by immunoblotting The RC mice presented 25%
of reduction in body weight, and less adipose tissue mass. The p-tyr tub by insulin in RC mice was decreased in hypothalamus
compared to controls. In addition, the p-tyr tub by insulin in insulin receptor, Akt and FOXO1 were increased in RC animals. In
contrast, obese animals presented 20% of increase in body weight and adipose tissue mass. The p-tyr tub by insulin was inhibited
in hypothalamus of obese animals. Similarly, the phosphorylation of insulin receptor, Akt and FOXO1 were decreased. To
investigate the regulation of tub in adipose tissue in vivo, RC and obese mice had insulin injected in their cava vena and the
epididimal adipose tissue was extracted. As well as the hypothalamus, insulin increased the p-tyr tub in vivo of adipose tissue in
control animals. This regulation is impaired in HFD animals.
Conclusions:
In summary, these results suggest that tub is regulated both by insulin and leptin in hypothalamus and adipose tissue in vivo.
Thus, metabolic or energetic changes (RC and HFD) can modify the regulation of this protein.
Keywords: tub, hypothalamus, insulin, caloric restriction
Resumo:24-104
DIFFERENT MODELS OF OBESITY PROMOTE DISTINCT MODULATIONS IN SKELETAL MUSCLE FIBERS.
Barreiros, G. B. ; Correia, M. H. ; Silva, A. F. ; Lazzari, A. C. ; Paul, L. F. ; Mello, E. V. S. L. ; Lepri, E. R.
Departamento de Morfologia/Universidade Estadual de Maring, UEM
Objectives:
The aim of this study was to analyze the skeletal fibers of the gastrocnemius muscle of rats in different models of obesity: the
model of hyperlipidic diet (rat food handled) and hypothalamic injury (intradermal injections of monosodium glutamate - MSG).
The analysis was based on the histological differences observed in the glycolytic metabolism of different types of skeletal fibers.
We used 20 male Wistar rats. The MSG group received 4g/Kg body weight of monosodium glutamate in the neonatal period, the
control group of MSG (CMSG) received saline injections. The fat diet group (DH) received a diet high in fat, in the first 50 days
of life and the control group of fat diet hyperlipidic (CDH) received normal food ad libitum. At 90 days all the groups were
sacrificed. Gastrocnemius muscles were dissected and processed by the histochemical NADH-TR reaction, used to distinguish
different types of fibers. The data were subjected to analysis of variance (One-Way ANOVA) followed by post-Tukey test with
significance level of 5%. The DH group had a higher body weight compared with CDH group (p
Conclusions:
The high fat diet leads to increased body mass and affects the oxidative metabolism of skeletal muscles, increasing the proportion
of oxidative fibers (glycolytic). Obesity induced by monosodium glutamate MSG, possible causes hypothalamus lesions that
change hormone levels leading to an increase in the body weight. This hormonal change might have somehow affected the
differentiation and refurbishment of muscle fibers.
Keywords: skeletal fibers, obesity, hyperlipidic diet , MSG, histological analysis
Financial Support: CNPq-UEM.
Resumo:24-105
RESVERATROL HAS A POTENTIAL THERAPEUTIC EFFECT ON REDUCTION OF VISCERAL OBESITY,
DYSLIPIDEMIA AND HIGHER BLOOD PRESSURE IN OVERWEIGHED ADULT RATS PROGRAMMED BY
EARLY WEANING
Franco, J. G. 1; Lima, N. S. 1; Amaral, T. A. S. 1; Resende, . C. 2; Maia, L. A. 1; Oliveira, E. 1; Lisboa, P.

C. 1; Passos, M. C. F. 1; Moura, E. G. 1
2
Departamento de Farmacologia, UERJ
1
Departamento de Cincias Fisiolgicas, UERJ
Objectives:
Studies in human and animal models have shown that malnutrition during critical periods of neonatal life (gestation and lactation)
is associated with later metabolic disorders, such as hypertension, type 2 diabetes and obesity. Recently, we have shown that
early weaning (EW) with no use of pharmacological substances or maternal separation, programmed for obesity,
hyperleptinemia, and leptin and insulin resistance in adulthood. Resveratrol (Res), a natural phytoalexin found in grapes has
received attention through the link with the French paradox and later with its beneficial effects against most degenerative and
cardiovascular diseases. Here, we evaluated the effect of Res on visceral fat depots, serum lipid profile and blood pressure in
adult rats programmed by EW.
After birth, lactating Wistar rats were separated in: EW (early weaning) - dams were wrapped with a bandage to interrupt
lactation during the last 3 days of lactation, and C (control) - dams whose pups had free access to milk throughout lactation (21
days). At least 8 litters were used per group. At the 150th day, two EW offsprings from each litter were randomly assigned to one
of two groups: EW - rats received 0.5% (w/v) aqueous methylcellulose (vehicle) and EW+Res- rats received Res (30 mg/kg per
day for 30 days). The control group received the vehicle. At the end of experimental period, rats were killed in order to collect
blood and visceral fat mass (VFM) mesenteric, epidydimal, and retroperitoneal. Blood pressure was measured at 180 days by
the tail-cuff method using a Letica LE 5000 device. Differences were considered significant at p
Conclusions:
These findings suggest that resveratrol (30 mg/kg/day for 30 days) prevented the development of visceral obesity, dyslipidemia
and the increase in blood pressure in adult rats programmed by EW.
Keywords: metabolic programming, resveratrol, early weanning
Financial Support: Capes, Cnpq and Faperj
Resumo:24-106
CHANGES IN THE DIETARY FATTY ACID COMPOSITION DURING THE FIRST HALF OF PREGNANCY
AFFECT METABOLIC ADAPTATIONS DURING LATE PREGNANCY IN THE RAT
Fernandes, F. S. 1,2; Sardinha, F. L. D. C. 1; Tavares do Carmo, M. G. 1; Herrera, E. 2

1
INJC - Faculdade de Nutrio, INJC/UFRJ
2
Facultad de Farmacia, Univ. San Pablo CEU
Objectives:
To study the metabolic effects of different sources of dietetic Fatty Acid (FA) intake during the first half of pregnancy (from day
0 to day 12) in rats.
Five groups of Sprague Dawley rats received semi-synthetic diets witch the only difference was the FA sources: I) 9 % soy oil;
II) 9% olive oil; III) 8% fish oil + 1% sunflower oil; IV) 8% flaxseed oil + 1% sunflower oil ; V) 8% palm oil + 1% soy oil. From
day 12 to day 20 of pregnancy, all the animals received the same commercial diet. Rats from each group were killed at day 0, 12
or 20 of pregnancy. Were studied also virgin rats that received the same treatment of the pregnant ones. The most abundant FA
from the diets were: I: 18:1n-9 and 18:2n-6; II: 18:1n-9; III: n-3 FA and 18:1n-9; IV:18:2n-6 and 18:3n-3 and V: 16:0 and 18:1n9. Plasmatic glucose has decreased progressively in all the pregnant rats, unless in V that changed at day 20 and not day 12 (c,
pIV>III>II>I (I:55,222,87g; II: 57,033,20g; III: 58,381,65g; IV: 61,841,35g; V: 64,972,85g); the number and weight of
fetuses was lower in V (number of fetuses: I:14,170,87; II: 14,001,46; III: 14,171,11; IV:14,710,57; V: 12,501,26,
p<0,05).
Conclusions:
Moderate changes in the FA composition of the diet in the first half of pregnancy changes the metabolic adaptations and affects
the fetal development of rats.
Keywords: fatty acids, lipids, metabolism, pregnancy
Financial Support: CAPES; Convnio EADS/CASA y Universidad San Pablo CEU/Espaa
Resumo:24-107
EFFECT OF OVEREXPRESSION OF THE INDUCIBLE HEAT SHOCK PROTEIN 70 IN MOUSE ON SKELETAL
MUSCLE ATROPHY AND SUBSEQUENT RECOVERY
Nascimento, T. L. 1; Rodrigues, D. C. 1; Moriscot, A. S. 1; Davilla, F. W. 2; Mou, Y. A. 2; Tombe, P. D. 2;

Mestril, R. 2; Miyabara, E. H. 1
1
Instituto de cincias Biomdicas , USP
2
Physiology Department, Loyola University of Chicago, LUC
Objectives:
This work aimed to investigate the effect of the overexpression of the inducible heat shock protein 70 (HSP70i) in mouse on
skeletal muscle atrophy and subsequent recovery.
Methods: Wild type (WT) and HSP70i-overexpressing transgenic mice were subjected to hindlimb immobilization for 7 days or
for immobilization (7 days) and subsequent recovery (7 days). After the experimental procedure, the animals were weighed and
euthanized; the soleus and the extensor digitorum longus (EDL) muscles were removed, weighed, and stored for analysis.
Histological muscle cross-sections were generated in order to determine the myofiber cross-sectional area (CSA) and the
immunoexpression of the satellite cell marker neuronal cell adhesion molecule (NCAM). Contraction experiments were
performed in order to obtain the maximum tetanic force (MTC). Results: EDL muscle weight did not change in all groups.
Soleus muscle weight decreased in WT animals immobilized and immobilized and recovered respectively (25.7% and 22.4%,
respectively). In transgenic mice there was a reduction in soleus weight only in immobilized group (40%). The myofiber type I
CSA of EDL did not change in both WT and transgenic mice. The myofiber type II CSA of EDL from the immobilized group
reduced in both WT and transgenic mice (49.7% and 15.9%, respectively), but only the transgenic mice recovered the myofiber
CSA after release from immobilization. The myofiber type I and II CSA of immobilized soleus muscles reduced in both WT
(27.4% and 31.7%, respectively) and transgenic mice (17.2% and 31.7%, respectively) and those myofiber type I and II CSA of
immobilized and recovered soleus muscles from both WT and transgenic mice showed values similar to their control intact
muscles. The percentage of NCAM-positive satellite cells in immobilized and immobilized and recovered was reduced only in
WT mice in both soleus (56% and 69%, respectively) and EDL muscles (36% and 57%, respectively). EDL muscle did not show
alteration either in MTC or in fatigue protocol. Soleus muscles from WT mice had a decreased MTC in immobilized group
(54.7%), which returned to the control values in immobilized and recovered group.
Conclusions:
The preliminary results suggest that HSP70i improves skeletal muscle recovery from atrophy.
Keywords: ATROPHY, HEAT SHOCK PROTEIN 70, SKELETAL MUSCLE
Financial Support: FAPESP (2009/53528-7; 2009/53528-7).
Resumo:24-108
NEONATAL MALNUTRITION AND KINETICS OF NITRIC OXIDE PRODUCTION: A STUDY OF CELL
INFECTION IN VITRO BY STAPHYLOCOCCUS AUREUS
Morais, N. G. 2; Miranda, C. A. L. F. 1; Costa, T. B. 2; Almeida, T. M. 1; Andrade, M. A. 2; Castro, C. M.

M. B. 2
1
Laboratrio de Imunopatologia Keizo Assami - UFPE, UFPE
2
Departamento de Medicina Tropical/UFPE, UFPE
Objectives:
To study the kinetics of nitric oxide (NO) by alveolar macrophages (AM) after cell infection in vitro with Staphylococcus aureus
(SA) in groups of rats with neonatal malnutrition (NM) and a group of well-nourished rats (N).
Male Wistar rats (n=12) were breastfed by rats whose diet contained 8% and 17% of protein during lactation (MN and N group,
respectivelly). After weaning, both groups were administered a normoproteic diet. The AM were obtained after tracheostomy,
through the collection of bronchoalveolar lavage. After isolation of different cell types in type plaque Falcon, systems were
formed: (NC) negative control only composed by AM, (PC) positive control added with lipopolysaccharide and one test system,
SA, containing AM and 100L of a suspension of ATCC 29213. The systems were incubated at 37 C, humid atmosphere and 5%
CO2. Every two hours, totaling 24 hours of incubation, 100L were removed from the supernatant of cultures. The quantification
of NO was based on the Griess reaction, by determining the concentration of nitrate/nitrite. Statistical analysis used the Student t
test and Mann-Whitney assuming a significance level of p 0.005 (NC-10h: 0,6850,0994M/mL; SA-10h: 0,4960,2M/mL;
NC-12h: 1,0840,0864M/mL; SA-12h: 0,5580,213M/mL; NC-14h: 1,2070,0981M/mL; SA-14h: 0,5770,21M/mL; NC16h: 1,2530,0766M/mL; SA-16h: 0,8550,181M/mL; NC-18h: 1,5020,193M/mL; SA-18h: 0,8940,166M/mL; NC-20h:
1,3500,167M/mL; SA-20h: 1,036 0,211M/mL; NC-22h: 1,0370,0616M/mL; SA-22h: 1,0370,199M/mL;NC-24h:
1,1740,114M/mL; SA-24h: 1,0370,134M/mL).
Conclusions:
The model of neonatal malnutrition adopted extensively altered function of phagocytic cells, compromising the nitrosative stress
of macrophages. Moreover, Staphylococcus aureus developed some mechanism to reduce levels of NO, favoring his stay in the
macrophages.
Keywords: NEONATAL MALNUTRITION, NITRIC OXIDE, MACROPHAGES, STAPHYLOCOCCUS AUREUS
Financial Support: CAPES/PROPESQ
Resumo:25-061
OVEREXPRESSION OF PTEN REVERTS THE MALIGNANT PHENOTYPE OF HUMAN COLORECTAL CANCER

CELLS.
Moraes, P. D. S. ; Arajo, W. M. D. ; Robbs, B. K. ; Viola, J. P. D. B. ; Daz, J. A. M.

Instituto Nacional de Cncer- Diviso de Biologia Celular, INCa
Objectives:
Phosphatase tensin homologue deleted on chromosome 10 (PTEN) is a tumor suppressor, which is greatly impaired in many
cancer types. It was reported that PTEN overexpression could inhibit proliferation, migration and invasion of different cancer
cells, however the role that this event plays during the colorectal cancer (CRC) progression remains unknown yet. In the present
study we analyze whether PTEN overexpression can inhibit the malignant phenotype of CRC cells.
We used HCT-116 cells, a cell line derived from human colorectal carcinoma, which is highly invasive and metastatic, as CRC
model. The cells were transfected with the PTEN wild-type (pBabepuroL Flag HA PTEN) plasmid using retroviral vectors.
Western blot analysis confirmed the overexpression of PTEN in transfected HCT-116 cells. We observed that after PTEN
overexpression the migratory and proliferative potential were reduced, as see by using the crystal violet and wound-healing
techniques, respectively. Moreover, we observed that PTEN overexpression induced a decreasing in the phosphorylation state of
GSK-3&beta confirming the inhibition of the Wnt pathway, which is associated with tumor progression in colorectal cancer.
Conclusions:
We show that PTEN plays an important role in events related to progression of colorectal cancer. However, more studies are in
course to confirm these finding and define whether PTEN overexpression can be useful to reverse the malignant phenotype of
CRC cells.
Keywords: PTEN, colorectal cancer, HCT-116, transfection, overexpression
Financial Support: FAF, CAPES, INCa, FAPERJ
Resumo:25-062
EVALUATION OF APOPTOTIC EFFECTS OF CYCLIC IMIDES DERIVATIVES ON HUMAN ACUTE LEUKEMIA
CELL LINES.
Machado, K. E. ; Andreossi, H. M. S. ; Oliveira, K. N. D. ; Licnio, M. A. ; Nunes, R. J. ; Santos-silva, M.

C.
Objectives:
Recent studies have shown that some cyclic imides derivatives have demonstrated important results as potential antitumor agents
(Bioorg Med Chem; 16:8440, 2008; Eur. J. Med. Chem; 44:4674, 2009; Anticancer Drugs; 19:23, 2008). The objective of this
work was to investigate the cytotoxic effect of cyclic imides derivatives on K562 human acute myeloid leukemia cells and Jurkat
acute lymphoblastic leukemia cells.

Initially, a screening of eleven cyclic imides was realized by MTT (3-(4,5-dimethiazol-zyl)-2-5-diphenyltetrazolium bromide)
assay and the compound with the best cytotoxic potential, 1-phenyl-1H-pyrrole-2,5dione (1), was selected. Chemical structure
modifications were performed from compound (1), creating more five derivatives: 1-(4-methyphenyl)-1H-pyrrole-2,5-dione (2),
1-(3,4-dichlorophenyl)-1H-pyrrole-2,5-dione (3), 1-(4-nitrophenyl)-1H-pyrrole-2,5-dione (4), 1-(4-methoxyphenyl)-1H-pyrrole2,5-dione (5) and 1-(4-hydroxyphenyl)-1H-pyrrole-2,5-dione (6). These six compounds were tested for their cytotoxicity on two
human acute leukemia cell lines, K562 and Jurkat. Cells were incubated with increasing compounds concentrations (0.1-100 M)
from 6 h to 72 h, depending on the experiment procedures. Cell viability was assessed using MTT assay. Flow cytometry was
performed to analyze the compounds effects on cell cycle by propidium iodide staining and also to detect the induction of
apoptosis by Annexin V-Fluorescein Isothiocyanate (FITC) staining. Apoptosis was yet evaluated by fluorescence microscopy
with ethidium bromide and acridine orange staining. All compounds showed cytotoxicity in a time and concentration-dependent
manner. In K562 cell line, IC50 value (50% inhibitory concentration) in 24 hours was 29.88 M (+/- 0.82); 24.99 M (+/- 1.39);
29.81 M (+/- 1.43); 26.29 M (+/- 1.28), 30.42 M (+/- 1.46) and 30.62 M (+/- 1.37), to compounds (1), (2), (3), (4), (5) and
(6) respectively. In Jurkat cells, IC50 value was 18.41 M (+/- 0.43); 27.26 M (+/- 0.75); 19.96 M (+/- 1.69); 24.81 M (+/1.85); 22.06 M (+/- 1,10); 25.35 M (+/- 1.78), to compounds (1), (2), (3), (4), (5) and (6) respectively. The effect of these six
compounds on cell cycle demonstrated that all treated cells showed a significant increase in the proportion of cells in S phase in
both K562 and Jurkat. Flow cytometry analysis of annexin-VFITC showed a significant increase in the percentage of annexin V
positive cells after treatment with compound (2) (71.58 +/- 0.07), compound (3) (84.82 +/-3.5) and compound (5) (59.02 +/- 8.3)
on Jurkat cell line, and with compound (3) (54.6 +/- 0.65) and compound (5) (46.96 +/- 4.9) on K562 cell line. These results
confirm that the percentage of cells observed in sub-G0/G1 phase are undergoing apoptosis and are compatible with the results
obtained in the assessment of cell viability by MTT and with ethidium bromide/acridine orange method.
Conclusions:
The studied cyclic imides derivatives demonstrated a cytotoxic effect against both K562 and Jurkat cells. This effect occurs
through an apoptosis pathway and it is related to cell cycle arrest in S phase. The biological effects observed in the present study
for the cyclic imides suggests promising applications, mainly to compound (3) and compound (5). However, further studies are
needed to determine the mechanism of apoptosis induced by these compounds.
Keywords: Apoptotic Effects, Cyclic Imides, Human Acute Leukemia
Financial Support: none
Resumo:25-063
EXTRACELLULAR ATP MEDIATE CITOTOXIC EFFECT IN CELL CULTURE OF HUMAN CERVICAL
CARCINOMA.
Mello, P. A. 1; Beckenkamp, A. 1; Bertodo, D. 1; Bruno, A. N. 2; Wink, M. R. 3; Lenz, G. 4; Buffon, A. 1

1
Departamento de Anlises/ Faculdade de Farmcia, UFRGS
2
Instituto Federal de Educao, Cincia e Tecnologia, IFRS
3
Laboratrio de Biologia Celular, UFCSPA
4
Objectives:
Adenosine nucleotides may carry out a cytotoxic effect upon tumor cells depending on its concentration. Therefore, extracellular
ATP could play a toxic effect by a mechanism that involves apoptosis, through P2X7 purinoceptors. Moreover, a mechanism that
involves suppression on apoptosis and cellular adhesion is an important factor in cervical carcinogenesis. In this work we intend
to evaluate the effect of adenine nucleotides on the proliferation and death in SiHa human cell line (ATCC) derived from cervical
carcinoma.
Cell viability was assessed using the MTT assay. For that, 2.000 cells/well were seeded on 96 multiwell plates, were grown for
48h, treated with different concentration of ATP, ADP, AMP and adenosine and incubated for 2h and 24h at 37C. To evaluate
the time effect, 20.000 cells/well were seeded on 24 multiwell plates, were grown for 72h and treated with 5mM ATP for 24, 48
and 72h. After treatment, cells were counted in a hemocytometer. The MTT assay showed a significant reduction on cells
viability only when cells were treated with 5mM ATP (58%) for 24h, compared to control. There wasnt a significant reduction
on cells viability during a time incubation of 2h. ATP played a cytotoxic effect in a time-depend manner. The counter assay
showed a significant reduction in number of cells at times 48h and 72h, which was proportional at time of treatment with 5mM
ATP (54% and 19%, respectively).
Conclusions:
Adenine nucleotides show cytotoxic effect in cells culture of cervical cancer (SiHa), like demonstrated by ATP in this study.
Therefore, they can be considerated important in modulation of this disease and may present a future application as anti-cancer
drug therapy.
Keywords: Adenosine nucleotides , ATP, Human cervical carcinoma, P2X7
Financial Support: CNPq, FAPERGS, Propesq - UFRGS
Resumo:25-064
CARVACROL: ANALYSIS OF CYTOTOXICITY AND APOPTOSIS POTENTIAL IN LEUKEMIA CELL LINE HL60
Incio Jnior, A. L. A. ; Freitas, B. C. ; Silva, M. J. X. ; Dantas, B. B. ; Martins, G. V. F. ; Silveira, A. L. ;

Arajo, D. A. M.
Objectives:
Carvacrol is a monoterpene present in the essential oil fraction of oreganum, and used nowadays on a large scale in the food and
cosmetic industries. Studies involving the anticancer activity of this compound have shown that it has cytotoxic effects on
different tumor cell lines, but this activity is expressed differently according to the strain tested (Phytomed. 17:581, 2010). With
this purpose, the cytotoxic effect of Carvacrol was evaluated on HL-60 cell line and human peripheral blood mononuclear cells
(PBMC).
HL-60 cells (fifty thousand cells/well) and PBMC (two hundred thousand cells/well) were treated with Carvacrol (80-250 M)
for 24h. The cell viability was analyzed by the MTT and flow cytometry. To assess the type of cell death caused in HL-60, cells
were incubated with the Carvacrol (80-250 M) for 24 hours. Etoposide (2.5 M) was used as a positive control. To analyze the
cell cycle distribution and mitochondrial depolarization was used flow cytometry. Morphological alterations were determined by
acridine orange/ethidium bromide staining. The IC50 value found for the HL-60 line in MTT assay was 172.5 14,6 M.
Carvacrol was more toxic to HL-60 cells than PBMC (IC50>300 M). Analysis by flow cytometry showed a significant
reduction in cell viability at concentrations of 150 M (reduction at 6,47%1,53% in 250 M). The morphological changes
caused indicate, mainly, to occur apoptosis at all concentrations tested, despite a low level of necrosis was observed. Flow
cytometry analyses showed that Carvacrol induced G1 phase cell-cycle arrest. A significant mitochondrial depolarization
occurred only at a concentration of 250 M (9,52%2,17%). Consistent with these observations, the drug could effectively
induce apoptosis, whose levels are concentration dependent.
Conclusions:
Carvacrol was less toxic for normal cells (PBMC) in comparison whit HL-60 cells. The molecular mechanism of this cytotoxicity
in leukemia cells involves induction of apoptosis.
Keywords: APOPTOSIS, CANCER, CARVACROL, CYTOTOXICITY, HL-60
Resumo:25-065
EXPRESSION OF ESTROGEN RECEPTORS IN BREAST CANCER CELL LINES AFTER ANTI-ESTROGEN
TREATMENT
Aguiar, G. ; Ierardi, D. F. ; Jasiulionis, M. G.

Farmacologia/Universidade Federal de So Paulo, Unifesp
Objectives:
Until now, breast tumors present high incidence levels worldwide. In Brazil breast tumors are the most incident tumor among
women. The improvement and development of new techniques that help to detect the disease in an early stage and the discovery
of new biomarkers that can direct a better treatment are extremely urgent. Nowadays, one of the most used biomarkers to direct
the treatment is the estrogen receptors. Three estrogen receptors are known, two of them, ER and ER are members of nuclear
receptor family, while GPR30 is a G protein-coupled estrogen receptor. As the estrogen receptors are responsible for regulation
the transcription of many genes, including themselves, the goal of this study was analyze the expression of these receptors in the
breast tumor cell lines MCF-7 and SKBr3 after treatment with tamoxifen, fulvestrant, anastrozole, 17--estradiol, and the
epigenetic drugs 5-aza-2-deoxycytidine (5AzaCdR) and tricostatin A (TSA).
RNA extracts from the cell lines MCF-7 and SKBr3 treated or not with anti-estrogen drugs, 17--estradiol and demethylating
agent was obtained using TRIzol Reagent (Life Technologies USA) protocol. cDNA was synthesized by using the reverse
transcriptase enzyme Superscript III (Invitrogen). The expression of the three estrogen receptor genes was carried out by Real
time PCR (qPCR). The GAPDH gene expression was used as a control of constitutive expression. According to American Type
Culture Collection (ATCC), the SKBr3 is a cell line derived from a metastatic mammary adenocarcinoma and shows tumorigenic
capacity. The MCF-7, despite of coming from a metastatic mammary adenocarcinoma, it retains characteristics of differentiated
mammary epithelium including ability to process estradiol via cytoplasmic estrogen receptors. The SKBr3 cell line, that is known
as not expressing ER, presented increased expression of ER after the treatment with tamoxifen (p
Conclusions:
These data suggest that the anti-estrogen drugs and the epigenetic drugs modify the expression pattern of estrogen receptors
analyzed in MCF-7 and SKBr3 cell lines. Nevertheless, more detailed analysis have to be done to confirm and validate these data,
but it already shows that these drugs influence the expression of estrogen receptors and that this can be in some grade related to
the response to treatment in breast tumor patients.
Keywords: breast tumor, estrogen receptors, anti-estrogenic treatment, expression analysis, cell line
Financial Support: Fapesp, CAPES, CNPq
Resumo:25-066
8-METHOXYPSORALEN INHIBITS GST-PI AND PRESENTS IN VITRO ANTI-GLIOMA ACTION
Oliveira, D. M. 1; Teles, A. L. B. 2; Farias, M. T. 3; El-bach, R. S. 1

Laboratrio de Neuroqumica - Universidade Federal da Bahia , UFBA
2
Laboratrio de Modelagem Molecular, UEFS
3
Laboratrio de Toxicologia, HSR
Objectives:
Brain tumors have been studied for a long time, nevertheless the impact of technological advances on clinical outcome has not
been satisfactory. The main reason for the unsuccessful therapy is chemoresistance. Among factors responsible for this resistance,
enzymes involved in drug metabolism have important role. The enzyme glutathione S-transferase-Pi (GSTP) promotes
conjugation of drugs with reduced glutathione (GSH) and its expression is up-regulated in a number of tumors, including brain
tumors. Then, a compound that inhibits GST-Pi activity has potential chemo-sensitizer action, and it would be useful in the
treatment of many types of cancer. Aim: This study investigated the GST-Pi inhibitory activity of the coumarin-derivative 8methoxypsoralen (8-MOP) by in vitro and in silico approaches.
A docking calculation of the 8-MOP at the active site of the GSTP1-1 (PDB structure 3IE3[1]) was carried out using the
Autodock Vina software and showed a binding energy of -6.4 Kcal/mol, against -6.1 and -5.9 Kcal/mol from two GSTP
inhibitors (ethacrynic acid and chlorambucil, respectively). Enzymatic kinetic analysis was performed in vitro using GSH and
chloro-dinitrobenzene (CDNB) as substrates. The reaction rate was determined spectrophotometrically at 340 nm. IC50 value
was calculated by non-linear regression increasing the concentrations of 8-MOP. Vmax and Km (mol/min.mg and mM,
respectively) were calculated following the Michaelis-Mentem pattern. 8-MOP inhibited dose-dependently GSTP activity (IC50
of 0.09 mM) through competitive way (it increased Km but did not change Vmax). Interestingly, 8-MOP acted as competitive
inhibitor, but not as substrate, since HPLC, and UV-Vis spectrophotometric analysis showed no 8-MOP-SG conjugate. 8-MOP
also inhibited GST activity in tests with extract of human glioma cells (GL-15). Living cells (in culture) were also used in this
work: monochlorobimane (MCB) assay was used to detect GSH in GL-15 cells and showed that 8-MOP does not deplete GSH,
but CDBN (GST substrate) does, confirming ours conclusions. As additional effects, 8-MOP treatment in vitro reduced tumor
cell proliferation (accessed by MTT assay, trypan blue exclusion test, and phase contrast microscopy) and promoted apoptosis
(accessed by annexin-V and propidium iodide staining/flow cytometry) and changes on cell morphology (accessed by scanning
electron microscopy). The same effects were observed in rat glioma cells (C6), but normal astrocytes were not affected to the
same extent as tumor cells were.
Conclusions:
8-MOP is a recognized effective drug for skin diseases, but we demonstrated, for the first time, its GST inhibiting potential, that
points out this substance as a possible chemo-sensitizer in cancer therapy. Data also suggest 8-MOP as a prototype for new anti-
cancer design, since it demonstrated direct and selective action on glioma cells.
Keywords: antiproliferative, cancer, glioma, gst, psoralen
Financial Support: Capes and CNPq
Resumo:25-067
DNMT3B OVEREXPRESSION CONTRIBUTES TO ABERRANT METHYLATION IN BURKITTS LYMPHOMA
CELL LINES
Arruda, V. O. 1,2; Robaina, M. C. S. 1,2; Klumb, C. E. N. P. 1,2

Laboratorio de Hemato-Oncologia Celular e Molecular, INCA
2
Programa de Ps-graduao em Oncologia , INCA
Objectives:
Burkitt Lymphoma (BL) is an agressive B-cell malignancy accounting for 80% of childhood and less than 5% of adult nonHodgkin lymphomas worldwide. Genetics and epigenetics alterations contribute to tumor progression and inactivation of the
apoptosis pathways in these tumors. Aberrant DNA methylation is the most widely studied epigenetic modification and is
associated with gene silencing. However its role in the pathogenesis of BL is still poorly elucidated. In the current study, we
analyzed three BL cell lines for expression of Bcl-2 pro-apoptotic family and DNA methyltransferase genes. Moreover, promoter
methylation patterns were analysed and correlated with gene expression status.
BL cell lines Daudi, Raji and BL41 were propagated in growth medium RPMI and harvested for RNA preparation, cDNA
synthesis (Read-to-go T-primed First-Strand Kit) and analysis of BIM and PUMA gene expression by quantitative Real Time
PCR using Sybr Green detection system. Gene expression levels were normalized using GAPDH for each cell line and
differences in the gene expression were determined using the comparative Ct method. All cell lines showed basal BIM expression
levels, as well as in the healthy donors sample. Low expression levels of PUMA were detected in Daudi and Raji cell lines.
Then, the expression of DNA methyltransferases enzymes (DNMT1 and DNMT3b) were analyzed by quantitative Real Time
PCR. An increased expression of DNMT1 about 2-fold higher was found in the BL41 cell line. Furthermore, an increase about
10-fold of DNMT3b expression levels were observed for BL41, and 7, 2.7-fold for Daudi and Raji cell lines, respectively. To
investigate whether the low gene expression was related to DNA methylation we performed a methylation-specific PCR (MSP).
The DNA was extracted using QIAamp DNA Blood Mini Kit (Qiagen) and converted with sodium bisulfite (Epitect Bisulfite Kit
Qiagen). The PCR assay was performed using MSP primers directed to specific segments within the promoter regions of selected
genes. All cell lines were methylated for the gene BIM and for the gene PUMA only the cell lines Raji and Daudi were
methylated.
Conclusions:
BL cell lines showed a decrease of BIM and PUMA gene expression, as well as DNA methyltransferase 3b is significantly
elevated in these cell lines. These findings suggest that the mechanism that accounts for the methylation-dependent gene silencing
observed is associated with the elevated DNMT activity, specially the overexpression of DNMT3b.
Keywords: METHYLATION , BURKITT LYMPHOMA , BIM, PUMA , DNMT3b
Financial Support: INCT para Controle do Cncer, CNPq 573806/2008-0; FAPERJ E26/170.026/2008
Resumo:25-068
POLYMORPHIC ABCB1 GENE STATUS, AND P-GLYCOPROTEIN EXPRESSION AND ACTIVITY DO NOT
INTERFERE WITH THE PTEROCARPANQUINONE- LQB-118-INDUCED APOPTOSIS IN CELLS FROM
MYELOID LEUKEMIA PATIENTS
Matta, R. R. 1; Scheiner, M. A. M. 1; Silva, L. F. R. 1; Vasconcelos, F. C. 1; Silva, A. J. M. 2; Costa, P. R. R.

2
; Maia, R. C. 1
1
Laboratrio de Hemato-Oncologia Celular e Molecular, LHOCM/INCA
2
Laboratrio de Qumica Bioorgnica, LQB/UFRJ
Objectives:
Multidrug resistance (MDR) is mediated by the efflux pump protein P-glycoprotein (Pgp) activity. Polymorphisms (SNPs) in the
ABCB1 gene may influence Pgp expression and activity. The synthetic compound pterocarpanquinone-LQB-118 has showed
anti-tumoral activity against myeloid leukemic cells. The objective is evaluate the influence of ABCB1 gene (exons 12 and 26)
SNPs in the apoptosis rate induced by LQB-118 in cells from myeloid leukemia patients.
Forty-three white peripheral blood from myeloid leukemia patients were isolated by Ficoll-Hypaque gradient and stocked in
Trizol, for DNA extraction. The genotypes of each ABCB1 gene variants, exons 12 and 26, were performed by PCR-RFLP assay.
Expression and activity of Pgp were performed by flow cytometry (FC). The apoptosis rates of leukemic cells were assessed
using the Annexin V assay by FC, after treatment with LQB-118 3.0 M for 48h. For the apoptosis rate analysis were considered
two groups: 20% (low apoptosis rate) and > 20% (high apoptosis rate). Frequency for exon 12: CT=28/43 (0.65), CC=12/43
(0.28) and TT=3/43 (0.07). For exon 26: CT=24/43 (0.56), CC=11/43 (0.25) and TT=8/43 (0.19). The patient group with high
apoptosis rate, 16/21 (76%) presented positive Pgp expression, and 20/21 (95%) positive Pgp activity. For positive Pgp
expression, 10/16 (63%) was CT for both exons. For positive Pgp activity, 12/20 (60%) was CT for exon 12 and 11/20 (55%) was
CT for exon 26. In the group with low apoptosis rate, 6/11 (54%) presented positive Pgp expression and 11/11 (100%) positive
Pgp activity. For positive Pgp expression, 3/6 (50%) were CT for both exons. For positive Pgp activity, 5/11 (46%) were CT for
both exons. Comparing the genotypic frequency in both exons for each group of apoptosis rate, there was not relevant difference,
except for the genotype TT in which there was an increase of the frequency of the samples exhibiting Pgp expression and activity
associated to low apoptosis rate.
Conclusions:
The genotype CT was the most frequent and TT the least frequent within the both exons of ABCB1 gene. In the group with
positive Pgp expression and activity, the genotype CT for exons 12 or 26 was more frequent, but this status did not indicate a
relation with the LQB-118-induced apoptosis. LQB-118 was capable to induce apoptosis in myeloid leukemia cells,
independently of the Pgp expression and activity status, but it seems related to TT SNPs status for exons 12 and 26 of the ABCB1
gene.
Keywords: leukemia, polymorphism, P-glycoprotein, ABCB1 gene, pterocarpanquinone-LQB-118
Financial Support: Swiss Bridge Foundation, INCT, FINEP, CNPq
Resumo:25-069
EVALUATION OF THE IMMUNOHISTOCHEMICAL EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH
FACTOR (VEGF) IN GASTRIC CANCER
Eifler, L. S. 1,4; Forgiarini, L. F. 4; Toneto, M. G. 4; Marroni, N. P. 1,2,3,4

1
Programa de Ps Graduao em Medicina , PPGCM/UFRGS
2
Programa de Ps Graduao em Fisiologia, PPG Fisiologia/UFRGS
3
Programa de Ps Graduao em Gentica e Toxicologia, PPGGT/ULBRA
4
Laboratrio de Fisiologia Experimental e Gastroenterologia, LFEG/HCPA
Objectives:
Evaluate the prognostic significance of VEGF expression and its correlation with clinicopathological presentations in gastric
resected tumors.
Methods: Cohort study with contemporary component, including 48 samples of human gastric tumors resected between 1995 and
2003. Tumors were prepared in paraffin blocks and VEGF expression evaluated by immunohistochemical study using rabbit
polyclonal A-20 (Santa Cruz Biotechnology,USA). Expression quantification was made by computer analysis identifying the
percentage and intensity of VEGF expression.The histopathologic staging used was the TNM International classification
proposed by the UICC (International Union Against Cancer) considering gastric wall penetration, lymph node involvement and
metastasis occurrence. Statistical analysis was performed by ANOVA followed by the Student Newman-Keuls test considering
the significance level p < 0.05. Results are presented as mean standard error. This study was evaluated and approved by the
ethics committee and research at Porto Alegre Clinicas Hospital. Results: We found significant differences (p
Conclusions:
In this study there was an inverse correlation in the intensity of VEGF expression with progression, staging and prognosis of
gastric tumors. The finding is likely due to a greater degree of hypoxia tissue in the tumor inducing increased angiogenic stimulus
and VEGF expression in the early stages of gastric tumors.
Keywords: VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF, GASTRIC CANCER, EXPRESSION
Financial Support: Fundo de Incentivo Pesquisa e Eventos FIPE
Resumo:25-070
THE LQB 118 INDUCE APOPTOSIS BY INTRINSIC OR ENDOPLASMIC STRESS PATHWAY
Bacelar, T. S. 1; Netto, C. D. 2; Costa, P. R. R. C. 3; da Silva, A. J. M. 3; Rumjanek, V. M. 1

1
Departamento de Qumica Orgnica, IQ

Ncleo de Pesquisas de Produtos Naturais, NPPN
Objectives:
Leukemias are characterized by accumulation of anormal undifferentiated cells in the bone marrow replacing healthy blood cells.
Treatment consists mainly of chemotherapy cycles, which may lead to selection and expansion of resistant cells. Multidrugresistance (MDR) phenomenon is the most important reason for failure during cancer chemotherapy. The extrusion of
chemotherapics mediated by ABC family transporters, overexpression of anti-apoptotic proteins such as Bcl-2, and p53 mutation
are classic mechanisms responsible for the MDR phenotype. Therefore, the development of novel chemotherapic agents against
MDR leukemias is of great interest. Our group tested different compounds with naphtoquinone and pterocarpan moieties, named
pterocarpanquinones being the most important, the compound named LQB 118. LQB 118 was efficient for different leukemia
cell lines and the objective for this work was to assess the pathways of cell death induced for this compound.
Cells K562 and Jurkat were grown in RPMI medium with 10% serum at 2x104 cells/mL or 1x105 cells/mL, and incubated with
the diverse compounds in concentrations varying from 1.25M to 50M. For apoptotic and mithocondrial membrane potential
(MMP) assay, cells were incubated in 3M of the compound and then, labeled with annexin-V/Pi (for apoptotic assay) or Dioc6
(for MMP assay). For caspases -9 and -12, cells were incubated in 3M or 8M of the compound and then, labeled with caspase9 or -12 staining kit. For the assay of intracellular calcium, cells were pre-incubated with Fluo-3 AM and then, incubated with
3M or 8M of the compound for 10min to 60min plus BAPTA AM or EDTA. Cell viability was measured by MTT after 72h.
Compound tested showed EC50 values around 3.5M against leukemia cells. The compound caused apoptosis and changes in
MMP and/or caspase-12 activation at 24h, and activation of caspase-9. The compound also was able to induce mobilization of
cytoplasmic calcium.
Conclusions:
According to data, this new synthetic compound exhibited cytotoxic effect in leukemic cell lines. The compound also induced
cell death for apoptosis by intrinsic or endoplasmic reticulum stress pathways. This work allowed us to demonstrate that the same
compound is able to activate different pathways of cell death according the dose or cell line used.
Keywords: LQB 118, leukemia, apoptosis, calcium
Financial Support: CNPq, FINEP, FAPERJ
Resumo:25-071
CORRELATION BETWEEN HOMOCYSTEINE CONCENTRATION IN SERUM AND RESPONSE TO ANTIHORMONAL TREATMENT IN BREAST TUMOR PATIENTS
Ierardi, D. F. 1; Germano, P. B. R. M. 1; Raimundo, L. G. 2; Mattar, A. 3; D'almeida, V. 4; Calegari, B. 4;

Porto, C. S. 1; Gebrim, L. H. 3; Correa, M. 2; Jasiulionis, M. G. 1
1
Farmacologia/Universidade Federal de So Paulo, UNIFESP
2
Instituto do Cncer do Estado de So Paulo, ICESP
3
Hospital Perola Byington, Hospital Perola
4
Psicobiologia/Universidade Federal de So Paulo, UNIFESP
Objectives:
Breast tumors are today the most incident tumor in women. One of the widely used markers for breast cancer is the estrogen
receptor (ER), which is member of the nuclear receptor superfamily of transcription factors. Almost 70% of the breast cancer
patients are ER positive and can be beneficiated with hormonal therapy. Patients with ER+ tumors are candidates for antihormonal therapy, including tamoxifen and anastrozol. Many studies proposed that elevated levels of total homocysteine (tHcy)
in the blood can be a risk factor for cancer as well as an important tumor marker. It is also known that estrogen status is
associated with tHcy concentration. One important epigenetic mechanism which is a well-described for carcinogenesis, and
seems to be related to homocysteine pathways, is the DNA methylation. In tumors, it is well documented a decrease in total DNA
methylation and a hipermethylation of specific gene promoters. There are no studies showing the impact of anti-hormonal
treatment in the tHcy concentration and in the DNA methylation pattern of the breast tumor patients. In this study we aimed to
verify which is the impact of the anti-hormonal treatment in the tHcy concentration and its correlation with the clinical data.
To investigate the effect of anti-hormonal therapy on homocysteine metabolism, 50 breast tumor patients were randomly assigned
to therapy with anastrozol or tamoxifen or placebo. Sera from the patients were obtained before and after the treatment with antihormonal drugs. Homocysteine levels were determined by HPLC. Total DNA methylation was evaluated in the tumor biopsies
before the treatment by restriction enzyme method. Pearsons correlation test and t test were performed for statistical analysis.
We observed that the tHcy before the anti-hormonal treatment increase in the patients with a high agressive tumor (p=0,0401),
and with positive lifonodes (p=0,0379). After the treatment with tamoxifen, the tHcy level decreases (p=0,0263). We could
observe that the decrease was significant in the patients with positive linfonodes (p=0,0400). No significant correlations were
observed when the patients were treated with anastrozol or placebo and also no correlation were observed in these patients when
we correlated the tHcy variation with the clinical data set. We observed a total DNA hypomethylation in the tumors before
treatment when compared with adjacent tissue (p=0,0157).
Conclusions:
We could observe that the higher tHcy levels are related to aggressive tumors. Our results also show that the treatment with
tamoxifen, mainly in aggressive tumors, decreases the tHcy levels showing that this treatment can have a role in the tHcy level.
Other clinical data and new studies should be done to evaluate the tamoxifen impact in tHcy and also its relation with the tumor
DNA methylation.
Keywords: Homocysteine, Breast tumor, Estrogen receptor, Tamoxifen, anti-hormonal therapy
Financial Support: Fapesp, CAPES, CNPq
Resumo:25-072
CHEMOSENSITIVITY OF THE SYNTHETIC COMPOUND LQB-118 THROUGH IAPS INHIBITION IN MYELOID
LEUKEMIAS CELLS
Castro, C. P. 1,3; Silva, K. L. 1,3; Reis, F. R. S. 1,3; Costa, P. R. R. 2; Maia, R. C. 1,3

1
Laboratrio de Hemato-Oncologia Celular e Molecular, INCA
2
Laboratrio de Qumica Bio Orgnica, UFRJ
3
Programa de Ps Graduao em Oncologia, INCA
Objectives:
Drug resistance is one of principal causes of treatment failure in myeloid leukemias. Although the treatment of chronic myeloid
leukemia (CML) has achieved major advances with the advent of imatinib, about a third of patients treated with imatinib need
alternative therapies, due to resistance or intolerance to this drug. Meanwhile, in acute myeloid leukemia (AML), the adult
survival rate after treatment with combination of an anthracycline and cytarabine is approximately 5% in five years. The
resistance to treatment and relapses are frequent events in AML patients. Overexpression of anti-apoptotic proteins such as the
members of inhibitor of apoptosis proteins (IAPs) Survivin and XIAP have been correlated to drug resistance and poor prognosis
in leukemias. In this context, the development of new drugs able to downregulate the expression of IAPs and hence facilitating
apoptosis is necessary. Previous results of our group showed that LQB-118, a synthetic compound was able to induce apoptosis
and downregulates the expression of Survivin and XIAP in a CML cell line. In the present work we evaluated whether LQB-118
was capable of inducing downregulation in CML and AML cells from patients and if it was correlated to apoptosis induction.
Leukemic cells from patients with AML (n = 7) and CML (n = 8) were treated with LQB-118 at concentrations of 3.0 M for 24
h. The levels of IAPs were determined by Western blot. Seven out of eight samples analyzed for Survivin expression, including
CML and AML, exhibited downregulation of this protein after LQB-118 treatment. In relation to XIAP expression, this
compound was capable of decrease the levels of protein in 8 out of 12 samples, while 4 samples had their levels of XIAP
increased. Besides, LQB-118 was able to induce significant apoptosis in the majority of leukemic samples, independently of
Survivin or XIAP overexpressions.
Conclusions:
Our results demonstrated that the compound LQB-118 is effective in inducing apoptosis through inhibition of Survivin and
XIAP. This finding suggests this drug as a potential good alternative in the treatment of drug resistance caused by these IAPs
overexpression in acute and chronic myeloid leukemias.
Keywords: MYELOID LEUKEMIAS, PTEROCARPANQUINONE, SURVIVIN, XIAP, APOPTOSIS
Financial Support: INCT, Fundao SwissBridge, FINEP, Prog. Oncobiologia (UFRJ/Fundao do Cncer)
Resumo:25-073
LQB-118 - A PTEROCARPANQUINONE - INDUCES APOPTOSIS THROUGH NFKB SUBCELLULAR
MODULATION AND IAPS INHIBITION IN CHRONIC MYELOID LEUKEMIA CELL LINES
Faria, F. C. C. 1,2; Silva, K. L. 1; Silva, A. J. M. 3; Costa, P. R. R. 3; Maia, R. C. 1,2

1
Laboratrio de Hemato-oncologia Celular e Molecular, INCA
2
Programa de Ps-graduao em Oncologia, INCA
3
Laboratrio de Qumica Bio-orgnica , UFRJ
Objectives:
Imatinib represents the most attractive and revolutionary therapy for BCR-ABL- positive oncoprotein in chronic myeloid
leukemia (CML). However, not all patients benefit from this type of treatment because of drug resistance phenomenon.
Consequently, research efforts have focused on developing more potent drugs with the ability to overcome the imatinib
resistance. In a previous study (Invest. New Drugs; 2010) we demonstrated that the treatment with the new synthetic compound
pterocarpanquinone named LQB-118 caused an important reduction of cell viability and increased apoptosis index in cell lines
derived from CML, both the vincristine-sensitive K562 cell line, and the resistant K562-Lucena overexpressing P-glycoprotein. It
was also observed the induction of caspase-3 activation, accompanied by a reduction of P-glycoprotein, survivin, and XIAP
expressions. The transcription nuclear factor B (NF-B) is related to oncogenesis through its capacity to regulate the expression
of a large number of genes that regulate apoptosis, cell proliferation and differentiation. Therefore, the aim of this study was to
evaluate which mechanism could be involved in the response observed after in vitro treatment with LQB-118 on both CML cell
lines K562 and K562-Lucena.
After treatment of the cell lines with each imatinib or LQB-118 we first evaluated the expression of Bim, antagonist of Bcl-2
family members, and Bcl-XL by Western Blot. Both cell lines treated with imatinib demonstrated increased levels of Bim, but the
same induction was not observed after LQB-118 treatment. The expression levels of the caspase-8 and IB, a NFB inhibitor,
were then analyzed by Western Blot. It was observed that the treatment with LQB-118 was able to induce the activation of
caspase-8 in both CML cell lines. Besides, LQB-118 treatment maintained and/or induced the levels of IB, suggesting that
NFB was kept inactive in the cytoplasm. These data were also complemented by immunofluorescence assay confirming the
NFB in the cytoplasm localization.
Conclusions:
Our findings indicate that LQB-118 induced high apoptosis index in the sensitive as well in the resistant CML cells through the
extrinsic pathway and modulated the activity of NFB, therefore preventing the expression of XIAP and survivin, reinforcing our
data in the previous study.
Keywords: NFkB, Chronic Myeloid Leukemia, IAP, PTEROCARPANQUINONE, Apoptosis
Financial Support: INCT, Programa de Oncobiologia (UFRJ e Fundao do Cncer)
Resumo:25-074
EFFECTS OF ABCB1 ACTIVITY ON MYELOID LEUKEMIC CELLS
Novis, B. F. ; Daflon-yunes, N. ; Rumjanek, V. M.

Objectives:
Multidrug resistance (MDR) is one of the main causes of chemotherapy failure. Tumor cells are capable of expressing transporter
proteins that are able to decrease the intracellular concentration of drugs used in cancer treatment, inducing the appearance of the
MDR phenotype in these cells. These transporters are members of the ABC transporter family. Among these proteins, ABCB1 is
probably the best known transporter linked to the MDR phenomenon. Some treatments of MDR tumors and other diseases use
ABCB1 inhibitors that may affect the progression of cancers that exhibit the expression of this transporter. Therefore, the aim of
this project was to analyze the effects of the different states of ABCB1 activity on myeloid leukemic cells. For this we studied
cells with ABCB1 activity inhibited (in the presence of its inhibitor), fully active (in the presence of its substrates) and basal
activity (in the absence of inhibitor or other substrates).
Cell lines used in this project were: K562, sensitive to chemotherapy; and FEPS, multidrug resistant cells selected from K562.
They were cultured with verapamil (ABCB1 inhibitor), with the chemotherapic ABCB1 substrate used to select FEPS cell line,
daunorubicin (DNR), or none of these drugs. Cells were kept in these experimental conditions for three days. Cell viability was
verified through MTT essay, and, the percentage of cells in each cell cycle phase was measured quantifying the nuclear DNA
using flow cytometry. Viable and non-viable cells were counted using Trypan Blue dye and light microscopy. Their ABCB1
expression was evaluated using a fluorescent antibody and the activity of the protein was analyzed through incubation with its
fluorescent substrate, both using flow cytometry. Results indicate that FEPS cells are resistant to DNR due to its expression of
functional ABCB1. FEPS cells kept in DNR presence showed a decrease in cell cycle rate and that may be due to the constant
activity of ABCB1 in this condition. FEPS cells kept in the presence of verapamil showed a decrease in its viability and that may
be a result of the inhibition of ABCB1. It was also shown a significant increase in ABCB1 expression when FEPS cells were kept
in presence of verapamil. The cells kept in all three conditions showed the same ABCB1 activity. That may be due to the increase
in ABCB1 expression in the presence of its inhibitor.
Conclusions:
These results suggest that the inhibition of ABCB1 may be compensated by its own overexpression, and that inhibition may also
decrease cell viability. Also, ABCB1 in constant activity may lead to a decrease in cell cycle rate.
Keywords: ABCB1, Myeloid Leukemic Cells, MDR, Cancer, Verapamil
Resumo:25-075
CELLULAR RESPONSE ANALYSIS IN COLORECTAL CANCER CELLS INDUCED BY ALQUILANT AGENTS IN
ENERGETIC STRESS CONDITIONS.
Lima, M1; Bordin, D1; Escargueil, A. 2; Larsen, A. 2; Soares, D. 1; Lenz, G1; Henriques, J. A. P. 1
1
2
Centre de Recherche Saint Antoine, UPMC
Objectives:
Solid tumours usually show regions with low vascularization and, as a consequence, these regions become hypoxic and deprived
of nutrients. In such conditions, a series of cellular functions are activated, which lead tumor cells not only to survive but also to
metastatic proliferation. Recently, it has been suggested that autophagy can contribute to the survival of tumor cells and conduct
to chemotherapy resistance. According to this, the proposal of this research was to analyze the cellular response of colorectal
cancer cells lines HCT116 wt and HCT116 p53-/-, treated with oxaliplatin (L-OHP) and cisplatin (CDDP) in low, normal and
high glucose conditions.
The cellular viability was determined by the MTT method that showed, for both drugs, a higher IC50 for cells treated in low
glucose, for both cell lines. The cell cycle, analyzed by flow citometry indicated that in low glucose conditions there was a G1
phase arrest, while higher glucose concentrations induced more cells to stop at S phase. The autophagy induction was determined
by flow citometry using acridin orange. Both drugs induced autophagy in HCT116 wt cell line, mainly in low glucose conditions.
No significant increase in autophagic cells was observed in HCT116 p53-/-.
Conclusions:
The findings suggest that these cell lines present a differentiated metabolism when exposed to low glucose which can contribute
to the survival of the cells. Besides, the results also suggest that p53 protein is involved in autophagy induction by L-OHP and
CDDP in colorectal cancer under low glucose conditions.

Keywords: alquilant agents, autophagy, colorectal cancer cells, low glucose
Financial Support: CNPq, FAPERGS, CAPES.
Resumo:25-076
MODULATION OF NAPI-IIB CO-TRANSPORTER ACTIVITY IN OVARIAN CANCER.
Sirtoli, G. M. 2,1; Neves, C. C. 1,3; Rangel, L. B. A. 2

1
Instituto de Biofsica Carlos Chagas Filho / UFRJ, IBCCF/UFRJ
2
Departamento de Cincias Farmacuticas/UFES, UFES
3
INCT-INBEB/CNPq/MCT, INCT-INBEB/CNPq/MCT
Objectives:
Ovarian cancer (OVCA) is the gynecologic malignancy with the higher mortality rate among women because the inefficiency of
early diagnosis, prognosis evaluation, and currently therapeutic strategies. Among the possible OVCA biomarkers, there is the
sodium-dependent phosphate co-transporter Type IIb (NaPi-IIb), codified by the gene SLC34A2, which has been already
associated to cancer development and/or progression. As a secondary active transporter, NaPi-IIb promotes the influx of sodium
and phosphate ions coupled to the electrochemical gradient generated and maintained by (Na+K+)ATPase. Similarly to the
observation for SLC34A2/NaPi-IIb, increased expression of 1- (Na+K+)ATPase has been documented in several carcinomas.
On the other hand, the enzyme inhibition has been related to the reduction in cellular proliferation and tumor invasion. Taken
together, the present work aimed to study the correlation of these transporters in 3 different OVCA lineages, A2780, ACRP e
OVCAR-3, respectively sensitive, acquired drug resistant phenotype, and refractory to cisplatin lineages.
Na+-dependent phosphate uptake by OVCA lineages was assayed by the difference of the transporter activity, expressed as nmol
Pi x min-1 x mg-1 protein, measured in the presence by the one obtained in the absence of Na2HPO4/NaH2PO4.H2O 0,1mM,
both in the presence of 32Pi 1mCi/mM. (Na+K+)ATPase activity measurement followed the method described by Grubmeyer &
Penefsky, and was expressed as nmol Pi x min-1 x mg-1 protein. Expression of (Na+K+)ATPase was analyzed by Western
Blotting. We have verified that the NaPi-IIb activity is approximately three fold higher in A2780 and four fold higher in ACRP
when compared to that of OVCAR-3 (A2780= 0,1410,015 nmol Pi x min-1 x mg-1; ACRP= 0,18670,023 nmol Pi x min-1 x
mg-1; OVCAR-3= 0,0460,005 nmol Pi x min-1 x mg-1;p
Conclusions:
Altogether, our data show an inverse correlation between the two transporters in OVCA lineages. The event is probably elicited
by a cellular mechanism in which the inhibition of (Na+K+)ATPase derives from phosphate accumulation due to the high activity
of NaPi-IIb. The elucidation of such mechanism might be the key to the understanding of NaPi-IIb role in OVCA. Therefore,
future experiments may lead to the discovery of molecular aspects related to ovarian malignant transformation, a condition that
still challenges global health by its impressive mortality rate.
Keywords: membrane transporters, NaPi-IIb, ovarian cancer, sodium pump
Financial Support: FAPES, FAPERJ, CAPES, INBEB, INPETAm and CNPq
Resumo:25-077
CYCLIN D1 EXPRESSION MEASURED BY REAL-TIME PCR IS DIFFERENT IN PATIENTS WITH MANTLECELL LYMPHOMA IN COMPARISON TO HEALTHY INDIVIDUAL
Paes, V. R. 1; Levy D1; Brocardo, G. A. 2; Pereira, J. 2; Bydlowski, S. P. 1

1
Laboratrio de Gentica e Hematologia Molecular , FMUSP
2
Laboratrio de Imunopatologia, HCFMUSP
Objectives:
Mantle cell lymphoma is an aggressive non-follicular small B-cell lymphoma that is associated with significantly shorter survival
(median survival is 3 years) despite a low-grade histology. Because of the aggressive nature of mantle cell lymphoma, accurate
diagnosis is important for prognosis and management. The t(11;14)(q13;q32) translocation causes deregulation of the bcl-1 gene
and over-expression of cyclin D1, which may in turn lead to lymphoma genesis. The bcl-1 translocation is specific for mantle cell
lymphoma and occurs in the majority of cases. The objective of this study was to evaluate the expression of cyclin D1 in healthy
human lymphocytes obtained from peripheral blood sample, by real-time PCR, to determine normal cyclin D1 expression.
5 mL of peripheral blood were collected from 21 healthy volunteers and 4 mantle-cell lymphoma patients after written informed
consent using EDTA as anticoagulant. Lymphocytes were isolated using Ficoll-Paque. Total RNA was extracted with Trizol.
Real-time PCR was performed using SuperScript III Platinum One-Step qRT-PCR Kit in a RotorGene RG-3000 (Corbett).
GAPDH was used as internal control. The primes and probes were bought from Applied Biosystems (cyclin D1:
Hs00765553_m1; GAPDH: 4333764F). PCR efficiency were evaluated using sequential dilutions from 500 to 31,25 ng of KG1
cell line. These concentrations were chosen because all the experiments had been done using 125 ng. Amplification of samples
was done in duplicate. The quantitative values were obtained from the cycle threshold (Ct), where the increase of signal is
associated to the PCR products exponential growth. The efficiency was calculated by the formula E = (10 -1/slope -1)X100. The
cyclin D1 efficiency was 98,13% and for the GAPDH 98,90%. RT-PCR results showed that the average expression of cyclin D1
gene in healthy individuals was 0,37 [0,14 0,59] and in mantle-cell lymphoma patients was 4,17 [1,10 7,23].
Conclusions:
Cyclin D1 expression in mantle-cell lymphoma patients measure by real time PCR was shown to be different from that in health
individuals. However, more studies are need to determine a cut-off value in order to use this technique for diagnostic tool.
Keywords: Cyclin D1, real time PCR, mantle-cell lymphoma
Financial Support: CNPq and INCT
Resumo:25-078
EVALUATION OF APOPTOTIC EFFECT OF CYCLIC IMIDES DERIVATIVES ON MURINE B16F10 MELANOMA
CELLS
Machado, K. E. ; Oliveira, K. N. D. ; Andreossi, H. M. S. ; Santos, L. ; Licnio, M. A. ; Nunes, R. J. ;

Santos-silva, M. C.
Objectives:
Cyclic Imides obtained by organic synthesis can be grouped in several subclasses, including maleimides, succinimides,
glutarimides, phtalimides, naphtalimides, and their respective derivatives. Due to their hydrophobic nature, these compounds can
cross biological membranes and exert important biological effects including anti-inflammatory, antinociceptive, antimicrobial
and antitumoral activities (Quim. Nova; 26:230, 2003). The objective of this study was to analyze the cytotoxicity of eleven
cyclic imides derivatives on Murine Melanoma Cells (B16F10).
Murine B16F10 melanoma cells were cultured in RPMI supplemented with 10% fetal calf serum, 100 U/ml penicillin, 100 g/ml
streptomycin and 10 mM HEPES, pH 7.4 at 37oC in a 5% CO2 humidified atmosphere. Cell viability was assessed using MTT
(3-(4,5-dimethiazol-zyl)-2-5-diphenyltetrazolium bromide) assay. Initially, a screening test evaluated eleven cyclic imides to
select the compound with the best cytotoxic potential. This was done by using increasing concentrations (0.1-100 M) of cyclic
imides derivatives for 24h of incubation. In this selection, compound 2-benzyl-1H-benzo[de]isoquinoline-1,3(2H)-dione (4) was
chosen for being the most effective. Subsequently, cells were incubated with increasing concentrations (0.1-100 M) of
compound (4) for 24, 48 and 72h. Compound (4) reduced viable cells in a concentration-dependent manner when compared to the
control group (not treated cells), with an IC50 value (50% inhibitory concentration) of 77.75 M (+/- 1.3) for 24 hours, of 69.62
M (+/- 2.3) for 48 hours and of 38.99 M (+/- 3.0) for 72 hours. Flow cytometry was performed to analyze the compound
effects on cell cycle with the use of propidium iodide. Cell cycle results demonstrated that treated cells showed a significant
increase in the proportion of cells in sub-G0/G1 phase (18.76%), S phase (31.05%) and G2/M phase (21.8%), accompanied by a
significant decrease in G0/G1 phase (47.47%), when compared to the control group. Two methods were employed to determine if
cell death occurred by necrosis or apoptosis. One method used fluorescence microscopy with ethidium bromide and acridine
orange staining and other method used flow cytometry with Annexin V-Fluorescein Isothiocyanate (FITC) staining. The assay
using Annexin V-FITC Apoptosis Detection kit showed a significant increase in the percentage of annexin V positive cells
(69.27%) when compared to the control group (36.70%). These results confirm that the percentage of cells B16F10 observed in
sub-G0/G1 phase are undergoing apoptosis and are compatible with the results obtained in the assessment of cell viability by
MTT and with ethidium bromide/acridine orange methods.
Conclusions:
The studied cyclic imide derivative, compound (4), had a cytotoxic effect against murine B16F10 melanoma cells. This effect
occurred through an apoptosis pathway, and it is related to cell cycle arrest in S and G2/M phases. The cytotoxic effect observed
in the present study with the use of cyclic imides suggested promising applications. However, further studies are needed to
determine the mechanism of apoptosis induced by this compound.
Keywords: APOPTOTIC EFFECT, CYCLIC IMIDES, MURINE MELANOMA CELLS (B16F10)
Financial Support: none
Resumo:25-079
EVALUATION OF CYTOTOXICITY INDUCED BY LAURYL GALLATE ON K562 HUMAN ACUTE MYELOID
LEUKEMIA CELLS
Maioral, M. F. 1; Ferreira, S. C. 1; Licinio, M. A. 1; Silva, M. C. S. 1; Nunes, R. 2; Yunes, R. A. 2;

Creczinsky-pasa, T. B. 3; Rosa, G. R. 1
1
Depto. de Anlises Clnicas - Universidade Federal de SC, UFSC
2
Depto. de Quimica - Universidade Federal de SC, UFSC
3
Depto. de Cincias Farmacuticas -Universidade Federal de SC, UFSC
Objectives:
Recent studies have shown that gallic acid and its alkyl esters (methyl, propyl, octyl and dodecyl) induce apoptosis in different
cell lines (Arzneim-forsch.55:66, 2005; Leuk Res. 33:297, 2009). Since new compounds with biological targets and less
cytotoxicity to normal cells are necessary for cancer therapy, the aim of this study was to evaluate the cytotoxic effect of lauryl
gallate in human K562 acute myeloid leukemia cells.
The cell viability was determined by MTT colorimeter method. After incubation of K562 cells with increasing concentrations (1100M) of lauryl gallate for 12, 24 and 48 h, the compound reduced the percentage of viable cells in a concentration and timedependent manner when compared with the control (non-treated cells). The IC50 (50% inhibitory concentration) in 24h was
observed at a concentration of 30M. The greatest cell death was observed after 48 h and the percentage of viable cells was
92.48% +/- 3.86, 80.33% +/- 3.5, 56.80% +/- 2.16, 39.03% +/- 2.51, 25.55% +/- 1.72, 13.01% +/-0.47 and 4.86% +/- 0.31, for the
concentrations of 1, 2.5, 5, 10, 30, 50 and 100 M, respectively. The apoptosis induction was assessed by bromide and acridine
orange method and by Annexin V-FITC Apoptosis Detection kit. For the first method, the cells were treated with the compound
(30M) for 12, 24 and 48 h, marked with ethidium bromide and acridine orange and observed in a fluorescence microscope. This
method showed that lauryl gallate (30 M) caused apoptosis in K562 cells and that the number of cells stained with ethidium
bromide increased in 24 and 48 h, confirming the result that lauryl gallate reduces the percentage of viable cells in a time
dependent manner. The Annexin V-FITC Apoptosis Detection kit was used according to the manufacturers instructions. It
showed that 77.5% of K562 cells treated with 30 M of lauryl gallate expressed Annexin, compared with 7.2% of the control.
The cell cycle phase analysis was carried out by flow cytometry after propidium iodide staining. In comparison with the control
group, treated cells (30M of lauryl gallate) showed a significant increase in the proportion of cells in the subG0/G1 (40.55 +/0.184) and G0/G1 (50.98 +/- 1.58) phases, accompanied by a significant decreased in the G2/M (14.35 +/- 1.34) phase. All tests
were performed in triplicate.
Conclusions:
These results are in agreement with studies described in the literature since lauryl gallate showed a cytotoxic effect in K562 cell
line in a concentration-dependent manner. Moreover, it was demonstrated that cell death occurred by apoptosis rather than
necrosis. The evaluation of the cell cycle showed that lauryl gallate (30M) caused a block in G0/G1 phase and increased the
number of cells in apoptosis. This may indicate that the mechanism by which lauryl gallate inhibits the growth of leukemic cells
is not only by apoptosis but also by cell cycle arrest.
Keywords: apoptosis, cytotoxicity, K562 cells, lauryl gallate, leukemia
Financial Support: None.
Resumo:25-080
EFFECTS OF TUMOR-DERIVED EXTRACELLULAR MATRIX ON ENDOTHELIAL CELL FUNCTIONS:
IMPLICATIONS TO TUMOR-ASSOCIATED ANGIOGENESIS
Brando-costa; Saldanha-da-gama; Helal-neto; Morandi; Barja-fidalgo

Farmacologia e Pscicobiologia, UERJ
Objectives:
Tumor progression creates a microenvironment that influences neighboring cell plasticity, migration and invasion. Therefore,
tumors are reported to induce the formation of a new capillary network from preexisting blood vessels (angiogenesis). This
process depends on the production, by tumor cells, of growth factors. Adhesion to extracellular matrix is also important to
regulate endothelial cell survival, proliferation and motility during angiogenesis. Integrins are the major protein adhesion
molecules responsible for cell-matrix interactions, transducing molecular signals from microenvironment to regulate cell shape,
survival and proliferation. However the contribution of tumor matrix/endothelial cell interaction to tumor-associated angiogenesis
is poorly studied. In the present study, we evaluated the interactions between tumor-derived extracellular matrix and endothelial
cells and how these interactions may impact on tumor-associated angiogenesis
Extracellular matrix was obtained from a human melanoma cell line, MV3 and from a melanocyte cell line, NGM. To matrix
characterization, NGM and MV3 were cultivated for 24, 48 and 72 hours and the content of tenascin-C (TN), lamin (LN),
fibronectin, thrombospondin and collagen I and IV was evaluated by indirect ELISA. MV3 - derived matrix has significantly
more LN and TN than NGM-derived matrix. The contribution of each matrix on proliferation of a human endothelial cell line,
HMEC1, was evaluated by MTT assay. We observed that, after 48 h of incubation, HMEC1 proliferated more over MV3-ECM
when related to NGM-ECM. To investigate the contribution of specific integrins to endothelial cell proliferation we used a panel
of different disintegrins and ADAM 9D. Proliferation increase was inhibited by incubation with ADAM 9D, a ligand of laminin
receptor. Then, we investigated the role of ECM on adhesion, migration and tubulogenesis. HMEC1 adhesion on MV3 or NGMderived matrix was assayed by MTT. After 2 h, we did not verify any difference on the groups studied. To transmigration assay
we use a transwell plate where MV3 or NGM were cultured inside the inserts for 48 h before cell lyses. HMEC1 were seeded and
let to migrate through the matrix to a chemotatic stimulus (FBS) for 6h. MV3-derived matrix was able to increase endothelial cell
migration. Matrigel was used to study tubulogenesis by HMEC1. Previously, HMEC1 was cultured on MV3 or NGM derived
matrix for 24 h, then plated on a matrigel for 8 h. MV3-derived matrix was able to increase size and number of cell tubular
structures. Finally, we evaluated the effect of tumor-derived matrix on activity of metalloproteases by zimography gel. We
observed an increase in the activity of metalloproteases of HMEC1 cells grown on MV3 derived matrix.
Conclusions:
These data suggest that tumor-derived extracellular matrix influences endothelial cell functions and may play an important role in
tumor-associated angiogenesis.
Keywords: angiognese, clula endotelial, integrina , matriz extracelular, melanoma
Financial Support: Capes, CNPq and FAPERJ
Resumo:25-081
ANTIPROLIFERATIVE ACTIVITY OF PTEROCARPOQUINONES ON TUMOR CELL LINES
Lima, D. J. B. 1; Milito, G. C. G. 2,1; Costa-lotufo, L. V. 1; Moraes , M. O. 1; Buarque, C. D. 4,3; Pessoa, C.

1
; Costa, P. R. R. 4
1
Fisiologia e Farmacologia/ Universidade Federal do Cear, UFC
Fisiologia e Farmacologia/ Univ. Federal de Pernambuco, UFPE

Depto de Qumica/Pontifcia Univ. Catlica do Rio de Janeiro, PUC-RIO
4
Centro de Cincias da Sade/ Univ. Federal do Rio de Janeiro, UFRJ
Objectives:
Pterocarpoquinones possess biological properties such as antineoplasic activity, indicating that this chemical core could be an
interesting lead for new compounds. Then, the present work aims to investigate the cytotoxicity of new synthetic
pterocarpoquinones and azapterocarpoquinones.
Firstly, the antiproliferative effect of 9 compounds, LQB 149, 192, 205, 216, 221, 222, 223, 226 and 235 were evaluated by MTT
assay on leukemia (HL-60, K-562), glioblastoma (SF-205), colon (HCT-8), and melanoma (MDA-MB435) human cell lines after
72h of treatment. Only compounds that presented cytotoxic activity were assayed for hemolytic activity for 60 min using mouse
erythrocytes suspensions (2%) in NaCl (0.85%) and for cytotoxicity on peripheral blood mononuclear cells (PBMC) isolated
from human donors after 72h of incubation, by MTT assay. The MTT assays on tumor cells showed that 6 compounds, LQB 192,
216, 222, 223, 226 and 149 were capable to reduce in vitro tumor proliferation, being LQB 192 and LQB 223 the most active
compounds with lowest IC50 values at HL60 (0,4 and 0,3 M, respectively). Both compound presented low toxicity toward
PBMC (7,2 and 23 M, respectively). None of the six substances tested caused hemolysis, suggesting that cytotoxicity
mechanism is related to a more specific pathway.
Conclusions:
These findings indicated the potential of LQB 192 and 223 as new compounds with anticancer properties.
Keywords: PTEROCARPOQUINONES, antineoplasic , cytotoxicity , antiproliferative, MTT
Financial Support: CNPq, UFC
Resumo:25-082
LACK OF ASSOCIATION OF C677T MTHF-R (METHYLENETETRAHYDROFOLATE REDUCTASE)
POLYMORPHISM IN PATIENTS WITH DIFFUSE LARGE B CELL LYMPHOMA
Silva, K. S. 1; Maselli, L. M. F. 1,2; Chaves, D. S. C. 2; Bydlowski, S. P. 1

1
Cincias Mdicas/ Faculdade de Medicina da USP, FMUSP
2
Fundao Pr-Sangue Hemocentro de So Paulo, FPSHSP
Objectives:
The Diffuse Large B Cell Lymphoma (DLBC-L) is a heterogeneous clinicopathologic process that accounts for 60% to 70% of
cases of Non-Hodgkin's lymphoma (NHL) classified as aggressive. Its pathogenesis involves the accumulation of multiple
genetic events in different genes. Among the genes related to the development of various types of cancer many are polymorphic
as the gene of the enzyme methylenetetrahydrofolate reductase (MTHFR). This gene has a crucial role in folate metabolism being
important for the maintaining of the biosynthesis of DNA and/or RNA and in the methylation of genomic DNA. It is widely
accepted that the absence of these events lead to a genomic instability that may be critical in the oncogenic transformation of
human cells, favoring a possible link between MTHFR and carcinogenesis. The C677T change in MTHF-R gene promotes an
Alanin for Valin substitution in the protein, which results in a thermolabile enzyme with reduced activity. Whereas the MTHFR
C667T polymorphism could contribute to cancer risk (as observed for colorectal, lung, prostate cancer, etc). The purpose of this
study was to evaluate the MTHFR C667T polymorphism in patients with DLBC-L.
Genomic DNA was obtained from 5mL of whole blood from 129 patients with (DLBC-L). The MTHFR C667T polymorphism
analysis was performed by PCR/RFLP using the restriction enzyme Hinf I. The restriction product was analyzed in 2% agarose
gel stained with ethidium bromide. The allelic frequencies observed were C= 0.67, T= 0.33 and genotypic frequencies were 46%
for CC; 42% for CT and 13% for TT genotypes, respectively. The results obtained here indicate that the genotype frequencies of
MTHFR C667T polymorphism were not significant in this study group.
Conclusions:
No association was found between C667T genotype and DLBC-L due to the independent status of promoter methylation in
MTHFR gene or genomic ancestry of the patients.
Keywords: DIFFUSE LARGE B CELL LYMPHOMA, MTHF-R , POLYMORPHISM
Financial Support: Capes and CNPq
Resumo:25-083
EPIGENETIC CONTROL OF KEY REGULATORS OF THE EPITHELIAL-MESENCHYMAL TRANSITION AND
STEM CELL-LIKE PHENOTYPE ALONG THE GENESIS OF MELANOMA
Morais, A. S. ; Souza, C. F. D. ; Molognoni, F. ; Jasiulionis, M. G.

Departamento de Farmacologia - INFAR, UNIFESP
Objectives:
Introduction: Melanoma is the most aggressive form of skin cancer and its incidence is increasing worldwide. Although
melanoma is often curable when early diagnosed, the clinical extreme, metastatic melanoma, detains the worst prognosis and
remains one most resistant tumor types. In this way, continuous efforts have been made to better understand the molecular
pathogenesis of this disease with the aim to discovery new treatments to metastatic disease. It has been strongly suggested that
like mutations (genetic alterations), aberrant epigenetic events also take place in specific steps and are important to tumor
establishment and progression. Aim: Based on a murine model of melanocyte malignant transformation, the aim of our group has
been to identify and understand the effective participation of epigenetic mechanisms in the acquisition of a transformed
phenotype in our model.
Methodology: Gene expression profiles were analyzed through oligonucleotide microarray technology (GeneChip Mouse
Genome 430 2.0 Array, Affymetrix Inc.) and validated using RT- PCR and Real-time PCR assays (SYBR Green, Qiagen). In
order to demonstrate the role of epigenetic events in gene expression, the same assays were performed in cells treated or not with
demethylating agent, 5-Aza-CdR, and histone deacetylase inhibitor, Trichostatin A. Protein expression and subcellular
localization were evaluated by Western Blot and Immunofluorescence assays. Results and brief discussion: The model of
melanoma genesis used was established in vitro by submitting non-tumorigenic melanocytes (murine melan-a lineage) to
sequential cycles of anchorage blockade, suggesting that the interference of microenvironment may lead to intracellular
alterations with important gene expression impact. Our cell lineages represent distinct phases of melanoma genesis (melan-a
melanocytes; 4C, pre-malignant melanocytes; 4C11-, non-metastatic melanoma; 4C11+, metastatic melanoma). Moreover, it was
observed clearly morphological differences, mainly in intermediate stages of progression (in 4C and 4C11- cell lines), suggesting
a transient change between epithelial and mesenchymal phenotype along malignant transformation. This transient modification
was accompanied by the switch in the expression of master epithelial to mesenchymal transition (EMT) regulators, Twist1, Zeb1,
Wnt11, as well as Snail1. The presence of direct and indirect targets of Snail1, like &beta-catenin, E-cadherin and N-cadherin
was also evaluated and supports previous findings. In parallel, Chd1 (chromatin remodelling factor) and Nanog (pluripotency
factor) present the same expression pattern which may indicate that malignant transformation process followed by EMT and
MET (mesenchymal to epithelial transition) may be encouraged by an open chromatin configuration. Interestingly, expression of
most of these genes appear to be controlled by key epigenetic mechanisms, both DNA methylation and chromatin modification.
Conclusions:
Conclusion: We hypothesized that during melanocyte malignant progression associated with sustained stress, the occurrence of
an opened chromatin would facilitate cells to undergo changes in gene expression that allowing cell adaptation to new
environment and these alterations are orchestrated by epigenetic events
Keywords: epigenetic, EMT regulators, melanoma, cancer
Resumo:25-084
COMPARATIVE MORPHOLOGICAL ANALYSIS OF A SPONTANEOUS CANINE MAMMARY TUMOR IN
PRIMARY CELL CULTURE AND IN VIVO SPECIMENS.
Gouveia, G. M. 1; Paiva, M. B. 1,1; Soares-borges, J. C. 1; Oliveira, S. H. P. 1; Luvizotto, M. C. R. 1

1
1 departamento de clinica, cirurgia e reproduo veterinari , UNESP-Araatuba
2
2Departamento de Cincias Bsicas./ Faculdade de Odontologia, UNESP-Araatuba
Objectives:
In female dogs, mammary tumors are the most prevalent oncologic processes, comprising approximately 52% of all neoplasm.
These tumors have been used as a model for the human disease rather than murine assays, due to the common characteristics such
as histological and immunohistochemistry similarities, high incidence, heterogeneous nature of both canine and human
population, large number of recurrences and the absence of viral involvement in processs etiology . This study aims to present a
morphologic comparison of the histopathology, cell culture and cytology of a canine mammary tumor to verify the differences
between in vivo and in vitro neoplasm.
Mammary tumor samples were collected from a bitch during surgical procedure. A portion of the sample was fixed in 10%
formalin and imbedded in paraffin for histopathological evaluation with HE staining. Cytological smears were prepared using
fine needle aspiration biopsy (FNAB) technique (C1). For cell culture, a sample was collected and mechanically dissociated to
obtain explants that were placed in cell culture bottles with 2,5 mL of supplemented DMEM containing Streptomycin/Penicillin
(5000 UI/mL), Fungizone (250 g/ml), MEGS ( Mammary Epithelial Growth Supplement) and fetal bovine serum. The
bottles were storage at 37 C with 5% CO2 and 70% humidity. After the explants adherence, the medium was replaced every
other day until the cells reached confluence, and then transplanted to other bottles. At this moment, a sample was collected and
new smears were prepared using the same standard cytological protocol (C2). C1 showed predominance of pleomorphic
epithelial cells with round to ovoid nuclei associated with mild pleomorphic spindle cells with morphology of myoepithelial cells.
Cytological aspects were compatible with carcinoma in mixed tumor. Histopathology analysis confirmed the cytological findings,
with cartilaginous metaplasia. Smears from cell culture revealed bizarre and bulky epithelial cells with more malignant phenotype
and multiple coarse nucleoli. On the other hand, cells with myoepithelial features did not exhibit significant differences when
compared with C1. The cell culture protocol used was effective, demonstrated by the in vitro cell growing. Through
morphological analysis, it was possible to identify epithelial mammary cells both in histopathology and cytology, as in cell
culture, although significant increased malignancy criteria was noted in the last one.
Conclusions:
Cytological and histopathological analysis performed before and after cell culture provides comparative information of neoplasic
behavior, either in vivo or in vitro growth, and can be used as an important tool for researches concerning biological behavior of
canine mammary neoplasms.
Keywords: cell culture, dog, mammary tumor, cytology
Resumo:25-085
GERMLINE TP53PIN3 POLYMORPHISM IN HEALTH SUBJECTS FROM THE NORTH OF BRAZIL.
Paradela, L. S. 1; Paiva, S. 1; Mouro, M. M. 1; Montenegro, R. C. 1; Khayat, A. S. 2; Ribeiro-dos-santos, A.

1
; Assumpo, P. 3; Santos, N. P. C. 1; Santos, S. E. B. 1; Burbano, R. M. R. 2
1
Laboratrio de Gentica Humana e Mdica, LGHM / UFPA
2
Laboratrio de Citogentica Humana, LCH / UFPA
3
Hospital Universitrio Joo de Barros Barreto, HUJBB
Objectives:
TP53 is one of the major tumor suppressor gene, which is essential for the preservation of genome integrity. Different
poymorphic variants of the p53 gene have been widely investigated in population studies for their association with cancer
susceptibility. Intron 3 16 bp duplication polymorphism of TP53 (TP53 PIN3 - A1, non-duplicated allele; A2, duplicated allele)
has been reported to be associated with an increase cancer risk and has a strong modifier effect on TP53 germline mutations. This
polymorphism has been reported to be associated with a difference of several years (19 years) in the mean age at the first
diagnosis in TP53 mutation carrier. Thus, determination of PIN3 A2A2 genotype may provide a useful genetic marker in
predicatie high-risk individuals for the development of cancer and also for an early diagnosis. In this way, the aim of the study
was to evaluate the PIN3 Ins16bp polymorphism in healthy subjects in Belm, Par, the city in the Northern of Brazil with more
new cases of cancer in 2010.
High-quality blood samples for genetic studies were obtained from a series of 224 healthy brazilian and PIN3 Ins16bp
polymorphism were analyzed by PCR followed by sequencing in ABI 3130 using GeneMapper ID v.3.2. The Ethics Committee
in UFPA approved this work. In this study, 82.59% of the subjects have PIN3 A1A1, whereas 2.67% have PIN3 A2A2. Agestratification revealed that mutated allele is significant in young subjets (23.6 mean age) when compared to older subjects (38.9
mean age) (p< 0.0001). In a time point of 30 years-old, mutated allele was observed in 17% of the subjects under 30 years-old
and only in 2% in the subjects over 30 years-old. Consedering only mutated homozygose subjects, only 1 individual (0.9%) and 5
(4,3%) were found in subjects over and under 30 years-old, respectivily. A significant risk of 6 times to carry mutated allele
(PIN3 A1A2 or PIN3 A2A2) were observed in subjects under 30 years-old (OR=5.4; 2.14-13.9; IC95%;p=0,001) when compared
to older subjects.
Conclusions:
In our preliminar study, TP53 PIN3 germline polymorphism suggests a gene antecipation event in young subjects. A duplication
of intron 3 could increase the risk of developing cancer and thus this can be a genetic marker to early diagnosis of cancer,
however more studies and follow up of this subjects must be performed.
Keywords: cancer , genetic, polymorphism, PIN3, TP53
Financial Support: CAPES, CNPq, FINEP, FAPESPA
Resumo:25-086
NEW INSIGHTS OF P21WAF1/CIP1 ROLE IN MELANOMA PROGRESSION: ITS PARTICIPATION IN THE
CONTROL OF DNA METHYLATION
Colaneri, G. N. ; Cruz, A. T. D. ; Jasiulionis, M. G.

Departamento de Farmacologia- UNIFESP, UNIFESP
Objectives:
Aim: Cell cycle regulation, gene expression control mediated by epigenetic machinery and DNA repair mechanisms are events
tightly correlated. Disruptions in these pathways have a crucial effect in cell homeostasis balance and may contribute to
malignant transformation. p21waf1/cip1 was initially described as a cell cycle inhibitor. This protein is responsible for
modulating DNA repair and also seems to be able to interfere on DNA methylation. DNA methylation is correlated with
transcriptional repression. Such process is a consequence of methyl group addition on CpG dinucleotides, by DNA
methyltransferases (DNMTs). Involvement of p21waf1/cip1 in DNA methylation process is due to its competition with DNMT1
by the component of the replication fork PCNA. Thus, p21waf1/cip1 may influence DNA methylation profile and its deregulation
can favor tumor development. DNA methylation patterns are always altered in tumors, however, little is known about the
regulation of enzymes involved in this event. Several studies have shown that cell cycle components play a role in regulating the
expression and activity of DNMTs. Nevertheless, studies showing the role of p21waf1/cip1 in this regulation are rare. This
background combined with previous data obtained by our group prompted us to study the role of this protein in a model of
malignant transformation of melanocytes. Thus, the aim of this work is to assess the interactions involving p21waf1/cip1, PCNA
and the enzyme that maintains DNA methylation patterns, DNMT1 along melanoma genesis.
Methods and Results: In this study, we used a model of malignant transformation of murine melanocytes obtained through
induction of sustained stress. The cell lines utilized, which represent different stages in the progression of melanoma, were nontumorigenic melanocytes (ma), pre-malignant melanocytes (4C), non-metastatic melanoma (4C11-) and metastatic melanoma
(4C11+). Data from real-time PCR and Western blot showed changes in p21waf1/cip1 mRNA and protein levels, respectively,
over the transformation of melanocytes. The highest levels of expression were observed in 4C11- and 4C11+ melanoma cell
lines, and an even more significant increase was observed in 4C11+ metastatic cells. Protein immunoprecipitation assay was
performed to analyze p21waf1/cip1, DNMT1 and PCNA interactions. Results showed a differential association of these
components through melanocyte malignant transformation.
Conclusions:
Conclusion: The results obtained so far are an indication that, in this model, there are alterations in DNA methylation and that
these may be partly due to changes in the regulation of DNMT1 by p21waf1/cip1. Supported by FAPESP.
Keywords: cell cycle, DNA repair, epigenetic, melanoma
Resumo:25-087
LQB-118, A NOVEL ANTINEOPLASTIC AGENT, REDUCES B16F10 MELANOMA GROWTH AND INDUCES
THYMUS CELL SUBPOPULATION ALTERATION IN VIVO
Salustiano, E. J. 1; Dumas, M. L. 1; Netto, C. D. 2; da Silva, A. J. M. 3; Costa, P. R. R. 3; Rumjanek, V. M. 1

1
Instituto de Bioqumica Mdica, IBqM - UFRJ
2
Instituto Maca de Metrologia e Tecnologia, IMMT - UFRJ
3
Ncleo de Pesquisa de Produtos Naturais, NPPN - UFRJ
Objectives:
Cancer is a malignancy of difficult treatment and side effects of most antineoplastic agents contributes to therapeutic failure.
Therefore, development of novel, safer chemotherapy agents is of great interest. Among natural products with antineoplastic
effect, the pterocarpans, isoflavonoids able to induce DNA fragmentation, and the naphthoquinones, known for inducing
oxidative stress, were inspiration for a new hybrid synthetic molecule. LQB-118 was proven to be effective against human
leukemias and lung cancer in vitro on our previous works. Thus, this work aims to evaluate the effect of LQB-118 on the growth
of B16F10 murine melanoma in vivo. Safety was also considered, since in vivo toxicity was observed, highlighting effects on
immune system cells.
For toxicity evaluation of the compund, female swiss mice (6-8 weeks old and six months old) received a single, intraperitonial
acute dose of LQB-118 (3,8 mg/kg). After different periods (24 h, 72 h, 30 days and 90 days) weight alteration and behaviour
were observed (n=9). At the same time, thymus, spleen and bone marrow were excised and cells were analyzed by flow
cytometry to detect cell subpopulation alterations. Yet in this context, female C57BL/6 mice received daily intraperitonial
injections of LQB-118 (0,19 mg/kg/day) for two weeks in order to evaluate chronic toxicity (n=9). To evaluate the antineoplastic
effect of LQB-118, 105 cells B16F10 (murine melanoma) cells were subcutaneously injected on C57BL/6 mice, cells were left to
grow for three days and animals were then treated with daily intraperitonial injections of LQB-118 at chronic doses (0,19
mg/kg/day), for two weeks. Animals were euthanized, tumor mass was excised, and tumor size and weight were evaluated
(n=10). It was observed that LQB-118 did not show toxicity for young and adult mice since intraperitonial administration did not
seem to change weight gain, weight of the immune system organs and the absolute number of cells when compared to control
group. However, LQB-118 seems to lead to a decrease of T CD4+/CD8+ cells with concomitant increase of T CD4+ cells in
thymus. Furthermore, in vivo experiments showed that LQB-118 had an interesting antineoplastic effect in vivo, being able to
significantly reduce melanoma mass and size after two weeks.
Conclusions:
Data showed that the synthetic LQB-118 presents potential chemotherapy use on human patients, because of its low toxicity.
However, the effect of LQB-118 on T CD4 thymocytes still needs further investigation to understand its impact on the immune
system.
Keywords: Chemotherapy, Melanoma, Immunology, LQB-118, Thymus
Financial Support: CNPq, FINEP, FAPERJ, INCT - UFRJ
Resumo:25-088
EVALUATION OF THE SURVIVAL AND PROTEOLYTIC ENZYMES ACTIVITIES IN YOUNG TUMOURBEARING RATS SUBMITTED TO LEUCINE-RICH DIET AND VITAMIN-C SUPPLEMENTATION.
Duarte Junior, C. A. ; Gomes-marcondes, M. C. C.

Dept Physiology and Biophisics / Biology Institute, UNICAMP
Objectives:
Cancer is now responsible for the deaths of seven million people and eleven million new cases are revealed each year around the
world, most of these cases are characterized by metabolic changes characterized as cachexia. Thus, nowadays the study of coadjuvant therapies associated to traditional treatments is very important and become enhancing. Supplementation of leucine is
considered as an improving factor in many cases of cachexia, including those caused by the presence of tumor, because this
branched chain amino acid is considered the most important in muscle protein building. Meanwhile, the inhibition of oxidative
stress, caused by tumour growth, is applied to vitamin-C for its high antioxidant effect. The aim of this study was to determine
the effects caused by the association leucine-rich diet and vitamin-C supplementation on proteolytic enzymes activities in
gastrocnemius muscle of young Walker-256-bearing rats.
Weanling Wistar rats (21 days old, n=48) were distributed into eight groups, following standard criteria for nutritional support
and tumor implant. The rats received eutrophic diet (C) or leucine-rich diet (L) and/or were supplemented or not with vitamin-C
(VC) dissolved in drink water. The Walker-256 tumour-bearing rats received 2.5x105 viable cells in subcutaneous injection. In
all animals, the growth curve evolution was performed and measured the serum total protein, albumin and glucose, and the
enzymatic activities of cathepsin B and H, calpain, chymotrypsin-like and alkaline-phosphatase were assessed in gastrocnemius
muscle.The supplementation of leucine and vitamin-C (WLVC group) leaded to a longer survival even under the jeopardizing
effects of tumour growth (around 14 days) when compared to the animals without supplements (group W; survived 10 days). The
relative tumor weight showed 2% less in tumor mass in WLVC than group W. The serum albumin decreased in W (1.8 mg/dL)
animals compared to the controls (C 3,0 + 0,2 mg/dL ), although the lower serum albumin the WLVC group had slight decrease
as compared to W animals (WLVC 2.5 mg/dL). The enzymatic activity of chymotrypsin-like protease increased only in W group
(28.5 + 1,2 U//g/min) when compared to control and WLVC (19.2 + 0,3 and 20.5 + 1,0 U//g/min, respectively).
Conclusions:
The association leucine-rich diet and vitamin-C significantly improved the tumour-bearing rat survival, maintaining the
proteolytic activity pathways, as the control rats, suggesting a benefit of this association on reducing global cellular activities of
muscle tissue in cachectic animals.
Keywords: Cancer, CaChexia, Branched-chain amino acids, Ascorbic Acid, Proteases
Resumo:25-089
PHARMACOKINETIC AND METABOLISM INVESTIGATION IN RODENTS OF THE ACRIDINE ANTITUMOR
CANDIDATE AC04
Pigatto, M. C. 1; Ucha, F. D. T. 1; Torres, B. G. S. 1; Haas, S. E. 1; Lima, M. C. A. 2; Galdino, S. L. 2; Pitta,

I. D. R. 2; Lopes, N. P. 3; Costa, T. D. 1
1
Programa de Ps-Graduao em Cincias Farmacuticas/FACFAR, UFRGS
2
Depto. de Antibiticos, UFPE
3
Depto. de Fsica e Qumica, USP
4
Centro Bioanaltico de Medicamentos/FACFAR, UFRGS
Objectives:
AC04 (5-acridin-9-ylmethylene-3-(4-methyl-benzyl)-thiazolidine-2,4-dione) is an acridine derivative synthesized by the Ncleo
de Pesquisa em Inovao Tecnolgica (NUPIT, UFPE/Recife, Brazil) with important activity against solid tumors in mice.
Viewing this promising activity, the goal of this work was to investigate the pharmacokinetics of AC04 and its 1-oxo-AC04
metabolite disposition after intravenous (i.v.) dosing to Wistar rats.
Experiments approved by UFRGS Ethics in Research Committee (2008032 and 2007843). Awake male Wistar rats (n = 6)
weighting 250-300 g were injected with a single 1.5 mg/kg i.v. bolus dose into the lateral tail vein. After dosing, blood samples
were taken from the opposite tail vein up to 120 h. AC04 and metabolite were separated from plasma by deproteinization with
acetonitrile and quantified by previously validated liquid chromatography/mass spectrometry in tandem mode (LC/MS/MS)
method. AC04 individual plasma concentration-time profiles were adequately fitted to a two-compartment open model using
Scientist v. 2.01 (MicroMath) software and similar weight scheme. A vary rapid distribution phase followed by a much slower
elimination phase was observed. Pharmacokinetic parameters of AC04 were: clearance (CLtot) of 3.4 3.4 L/h/kg, volume of
distribution (Vdss) of 137.9 91.4 L/kg, area under the plasma concentration-time curve (AUC0-) of 788 483 ngh/mL and
terminal half-life (t1/2) of 45.5 31.5 h. A high interindividual variability on plasma profiles was observed, consistent with
preliminary studies. The rate of formation and elimination of 1-oxo-AC04, previously identified as the drugs main metabolite by
in vitro and in vivo studies, was determined. The mean 1-oxo-AC04 plasma profile obtained after AC04 i.v. bolus administration
showed that the metabolite is rapidly formed in vivo, presenting a t1/2 of 23.1 10.4 h. The metabolite reaches approximately
10% of the plasma concentration of the parent drug and whether it contributes to the parent drug activity or is potentially toxic
remains to be investigated.
Conclusions:
These results suggest that the AC04 distributes rapidly and extensively, presenting a large volume of distribution and plasma
half-life, which is important for its activity in solid tumors. The drug is metabolized forming 1-oxo-AC04 as main metabolite.
The set of results obtained indicate that AC04 has adequate pharmacokinetic properties and further pharmacological and
toxicological investigation should be conducted with this compound.
Keywords: pharmacokinetics, AC04, antitumor candidate, acridine derivative, LC-MS/MS
Financial Support: Financial support from INCT-if (CNPq/Brazil) and scholarships from CNPq/Brazil.
Resumo:26-157
EFFECT OF THE EXTRACT OF EUTERPE OLERACEA MART. (AAI) ON CARDIOVASCULAR CHANGES IN

SPONTANEOUSLY HYPERTENSIVE RATS
Cordeiro, V. S. C. 1; Carvalho, L. C. R. M. 1; Bem, G. F. 1; Sousa, M. A. V. 1; Sousa, P. J. C. 2; Soares de

Moura, R. S. 1; Resende, A. C. 1
1
Departamento de Farmacologia e Psicobiologia, UERJ
2
Universidade Federal do Par, UFPA
Objectives:
The Euterpe oleracea Mart. (aai) is a typical plant of the tropics, and rich in polyphenols. These substances have shown great
therapeutic potential, since it reduces the incidence of cardiovascular disease and its benefits may be associated with antioxidant
action, vasodilating and anti-hypertensive actions. The aim of this study was to investigate the protective effects of ASE in
arterial hypertension that is associated with an endothelial dysfunction and oxidative stress.
Methods:The experiments were approved by the Ethics Committee of Animal Experiments of the UERJ (protocol:
CEA/022/2010). Young male Wistar (W) and Spontaneously Hypertension Rats (SHR) (21 days old) received daily treatment
with ASE (200 mg/Kg/day) or not in water tap until 16 weeks old and systolic blood pressure (SBP, mm Hg) was measured by
plethysmography. The vasodilator effect of acetylcholine (ACh, 1-100 pmol) was studied in mesenteric arterial bed (MAB) precontracted with norepinephrine (30 M). Determination of oxidative damage was estimated by protein carbonylation (nmol/mg
protein) in kidney (K) and superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activities (U/mg protein)
in K and plasma (PL) by spectrophotometry. Results: SBP was increased in SHR compared to controls (SHR:2174.6 vs
W:1403.4 and W+ASE:1377.2) and treatment with ASE reduced the hypertension (SHR+ASE:1756.4; P0.05). The reduced
vasodilator effect (% of relaxation) of ACh (100 pmol) in SHR was recovered by the ASE (W:834.7; W+ASE:740.98;
SHR:593.7; SHR+ASE:851.8; P0.05). ASE decreased the levels of carbonyl protein in SHR (W:0.020.009;
W+ASE:0.040.011; SHR:0.070.014; SHR+ASE:0.0130,008; P0.05). The activities of SOD and CAT were reduced in K of
SHR compared to controls and treatment with ASE increased these activities (SOD: W:23.83.2; W+ASE:38.26.5;
SHR:7.63.5; SHR+ASE:46.216.9; CAT: W:0.180.034; W+ASE:0.140.052; SHR:0.020.009; SHR+ASE:0.100.027;
P0.05). The GPx activity was increased in K of SHR (W: 0.00060.0001; W+ASE: 0.00080.0001; SHR: 0.00220.0003;
P0.05) and decreased by treatment with ASE (SHR+ASE: 0.00130.0002). The activity of SOD in PL was not different
between groups (SOD: W:81.96.44; W+ASE:78.111.90; SHR:84.75.47; SHR+ASE:80.920.87). The antioxidant activities
of CAT and GPx were decreased in PL of SHR and ASE increased only GPx activity (CAT: W: 0.120.028;
W+ASE:0.120.030; SHR:0.040.012; SHR+ASE:0.070.009; GPx: W: 0.000920.00017; W+ASE: 0.000960.00017; SHR:
0.000350.00015; SHR+ASE: 0.000970.00017; P0.05).
Conclusions:
The results demonstrate that chronic treatment with ASE reduces the hypertension in SHR. The improvement of endothelial
function and the antioxidant activity induced by ASE may contribute to this beneficial effect of the extract in SHR.
Keywords: Aai, Endothelial dysfunction, Extract, Hypertension, SHR
Financial Support: CAPES and FAPERJ
Resumo:26-158
EFFECT OF THE ESSENTIAL OIL OF HYPTIS MARTIUSII BENTH. (LAMIACEAE) ON GASTRIC SECRETION
PARAMETERS AND GASTROINTESTINAL MOTILITY IN WISTAR RATS.
Caldas, G. F. R. 1; Silva, J. B. R. 1; Costa, J. G. M. 3; Wanderley, A. G. 2; Siqueira, N. C. 1

1
Departamento de Cincias Farmacuticas, UFPE
2
Departamento de Fisiologia e Farmacologia, UFPE
3
Departamento de Qumica Biolgica , URCA
Objectives:
Hyptis martiusii popularly known as cidreira-do-mato, grows in abundance in the Northeast of Brazil, where its leaves in infusion
or decoction has been used in folk medicine in the treatment of intestinal and stomachic diseases. The aim the study was to
evaluate the effect of essential oil from leaves of Hyptis martiusii (EOHM) on gastric secretion parameters and on gastrointestinal
motility in Wistar rats.
EOHM was obtained the dried leaves by hydrodistillation. The volume, pH and total acidity of gastric secretion were determined
by pyloric ligature model. The effects on gastric emptying and intestinal transit were also evaluated. Males rats (250-300g) were
used and divided into five groups (n=6) in either protocols. The animals were treated (i.d.) with EOHM (100, 200 and 400mg/kg),
vehicle (10mL/kg) and pantoprazole (40mg/kg) immediately after ligature of pylorus. In the gastric emptying, rats were
pretreated (p.o.) with EOHM, vehicle and atropine (3mg/kg, s.c.) 60 or 30 min before administration of the marker (phenol red, 5
mg/mL, p.o). The small intestine of animals used in the method of gastric emptying was removed for assessment of intestinal
transit. Data are expressed as means.e.m. Differences between groups were analyzed by ANOVA and Tukeys test (p
Conclusions:
These findings indicate that possibly the property the essential oil of the Hyptis martiusii of reducing gastric acidity and increased
pH, may play an important role on gastric mucosa protection. The continuity of studies with Hyptis martiusii will allow us to
scientifically validate the use of essential oil as a potential gastroprotective agent.
Keywords: ESSENTIAL OIL, Hytpis martiusii, GASTROINTESTINAL MOTILITY, GASTROPROTECTIVE
Resumo:26-159
CURCUMA LONGA L AND CURCUMIN ACUTELY ADMINISTERED SHOWS A POTENT ANTINOCICEPTIVE
ACTION IN UNRESTRAINED RATS
Carvalho, E. T. ; Souza-filho, J. O. C. ; Landim, M. A. ; Cruz, G. M. P. ; Viana, G. S. B.

Faculdade de Medicina Estcio de Juazeiro do Norte, FMJ
Objectives:
Curcumin (diferuloylmethane) is a major constituent of Curcuma longa L (turmeric), a medicinal Asian species widely used as a
condiment and in folk medicine due, among others, to its anti-inflammatory properties. Since the mechanism of its
antinociceptive effect is not fully understood, we decided to investigate the peripheral antihyperalgesic activity of both turmeric
and curcumin by a Plantar Test (Hargreaves Method) in unrestrained rats.
Male Wistar rats (180-200 g) were distributed into the following groups (10 to 12 animals per group): normal controls (NORM)
received only distilled water, p.o.; CARGN group, with animals that received water, p.o., and, subsequently, an intraplantar
injection of 100 L of 1% carrageenan (CARGN) into their right hind paw; INDO20, rats receiving Indomethacin (20 mg/kg,
p.o.); two groups treated with a turmeric (CL) aqueous suspension (100 and 200 mg/kg, p.o. CL100 and CL200, respectively)
and three groups treated with curcumin at the doses 10, 25 and 50 mg/kg, p.o. (CUMN10, CUMN25 and CUMN50). With the
exception of the NORM group, treatments of all the other groups were immediately followed by administration of CARGN. One
hour after the treatments, these animals were submitted to a focused thermal stimulation to provoke peripheral hyperalgesia. The
reaction time, in seconds, (by moving the injected paw) was measured. The Data were analyzed by ANOVA and the StudentNewman-Keuls test and expressed as means SEM (with significance when p< 0.05) and percentage reduction related to NORM
or CARGN groups. CARGN group showed a reaction time to thermal stimulation (6.31 0.57 s) 44% less than that of the
NORM group (11.23 0.46 s). Both turmeric and curcumin groups significantly increased the reaction time when compared to
CARGN. CL100 and CL200 increased the reaction time by 31% (8.24 3.49 s) and 53% (9.66 3.03 s) as related to CARGN,
respectively. The curcumin pretreatment showed a potent antinociceptive effect in this model for all doses tested. These results
were similar to that found with INDO20 (15.0 1.58 s), which showed an increased reaction time by 34% and 138% as related to
both NORM and CARGN groups, respectively. Thus, CUMN10, CUMN25 and CUMN50 increased the reaction times by 29, 53
and 46% (14.53 0.75, 17.15 0.90, 16.34 1.05 s) as related to NORM. In addition, these three curcumin doses increased
reaction times by 130, 172 and 159%, respectively, when compared to the CARGN group.
Conclusions:
Both Curcuma longa L and its most important constituent, curcumin, acutely administered significantly increased the reaction
time to thermal stimulation. Interestingly, at the lower dose tested a maximum effect of curcumin was already reached. This
observed antihyperalgesic effect was similar to that presented by a well-established anti-inflammatory drug, indomethacin. As
curcumin inhibits TNF- and NO releases as shown by others, these actions might be involved with the drug antinociceptive
effect.
Keywords: curcumin, antinociceptive effects of curcumin, Curcuma longa, Hargreaves Test
Financial Support: Faculdade de Medicina Estcio de Juazeiro do Norte - CE
Resumo:26-160
EFFICACY OF THE ESSENTIAL OIL OF PTERODON EMARGINATUS IN AN ANIMAL MODEL OF COMPLEX
REGIONAL PAIN SYNDROME-TYPE I (CRPS-I) IN MICE
Tomasini, L. 1; Nucci, C. 3; Mazzardo-martins, L. 1; Stramosk, J. 2; Santos, A. R. S. 1; Martins, D. F. 1,2,3

1
Departamento de Cincias Fisiolgicas, UFSC
2
Curso de Fisioterapia, Universidade do Sul de Santa Catarina, UNISUL
3
Curso de Naturologia Aplicada, Un. do Sul de Santa Catarina , UNISUL
Objectives:
Chronic post-ischemic pain (CPIP) is an animal model of CRPS-I developed using a 3-h. ischemia-reperfusion (IR) injury of the
rodent hindpaw. Complex regional pain syndrome, with major nerve injury (type II) or without (type I), is characterized by
spontaneous and stimulus-evoked pain, edema, vasomotor and sudomotor abnormalities, motor dysfunction, and trophic changes
(Pain 63:127, 1995). Pterodon emarginatus, is a native tree widely distributed over the central region of Brazil. Although an antirheumatic active compound seems to be present in their oleaginous fraction, seeds hydroalcoholic (wine) infusions are used in
folk medicine, throughout Brazil, for their anti-rheumatic, analgesic and anti- inflammatory properties. The present study was
designed to investigate the effect of the essential oil of Pterodon emarginatus (EOP) in a model of CPIP.
All experiments were conducted using male Swiss mice (25-35 g). Experimental procedures were carried out in accordance with
the National Institutes of Health Animal Care Guidelines (NIH publications No. 80-23), and were approved by the Ethics
Committee of the Universidade do Sul de Santa Catarina (UNISUL). Behavioral testing measurements were obtained from
separate groups of sham (n = 8), CPIP (n = 8) and CPIP + EOP (0.1-100mg/kg; intragastric, i.g.) (n = 8). Surgery: CPIP mice was
generated following exposure to prolonged (3 h.) hindpaw ischemia and reperfusion (IR). To assess hypersensitivity to
mechanical stimuli in CPIP mice, the right hindpaw was stimulated with 0.4 g von Frey filament (Stoelting, Chicago, USA)
(frequency response) (Eur. J. Pharmacol. 25:203, 2002). Two and seven days after CPIP, mice were treated with EOP (0.1100mg/kg, i.g.) and mechanical hypernociception was examined before and at different time-points after treatment (1, 2, 3 and 4
hours). In separate experiment, on day 7 after CPIP we evaluated the mechanical hypernociception 1h. after EOP (100mg/kg, i.g.)
at each day after CPIP for 5 days (7-11 days). On day 2 after IR, the frequency response to mechanical stimuli of CPIP mice, 1h.
after treatment with EOP, was significantly different from those of CPIP mice (p < 0.01); suggesting that EOP reversed
mechanical hypernociception (inflammatory pain) in CPIP mice. EOP (100mg/kg) administered by intragastric gavage inhibited
mechanical hypernociception. This action started 120 min. after i.g. administration (inhibition [I] of: 5511%) and remained
significant up to 3 h. (I= 4516%). On day 7 after IR, the frequency response to mechanical stimuli of CPIP mice, 1h. following
treatment with EOP, was significantly different from those of CPIP mice (p < 0.001), suggesting that EOP (100mg/kg, i.g.)
reversed mechanical hypernociception (neuropathic pain) in CPIP mice. EOP administered by intra-gastric (i.g.) gavage inhibited
mechanical hypernociception. This action started 60 min. (I: 4710%) after i.g. administration and remained significant up to 3 h.
(I: 494%). In separate experiment, daily treatment (7-11 days after IR) with EOP (100mg/kg, i.g.) decreased mechanical
hypernociception induced by CPIP, evaluated 1 h. after treatment (p < 0.01). This effect was evident until the fifth day of EOP
treatment (p < 0.001).
Conclusions:
The present results demonstrated that administration of EOP reduces mechanical hypernociception induced by CPIP in mice,
providing a rationale for its popular use in pain disorders.
Keywords: Chronic pain , CRPS-I, Neuropathic pain , Mice , Pterodon emarginatus
Financial Support: UFSC, Capes, UNISUL.
Resumo:26-161
EVALUATION OF THE HYPOTENSIVE EFFECT OF SYZIGIUM CUMINI
Herculano, E. D. A. 1; Feitoza, P. R. 1; Costa, C. D. F. D. 1; Paulino, E. T. 1; Aquino, P. G. V. 3; Frana, P.

H. B. 3; Arajo-jnior, J. X. 3,1; Santana, A. E. G. 3; Ribeiro, . A. N. 1
1
ESCOLA DE ENFERMAGEM E FARMCIA, ESENFAR
3
INSTITUTO DE QUMICA E BIOTECNOLOGIA, IQB
Objectives:
Syzigium cumini (Myrtaceae) is a plant popularly known as brinco-de-viva or jamelo. The specie has been described in the
literature as antifungal (J Antimicrob Chemother. 60; 312, 2007), anti-allergic (Braz J Med Biol Res 40; 1, 2007), anti-diabetic
(Planta Med. 9; 139, 1986), and anti-mutagenic (Phytochemistry. 36: 1027, 1994). The aim of the present study was to investigate
the acute cardiovascular effects of the ethanolic extract of the Syzigium cumini fruit (EESCF) in rats.
The extract was prepared by the percolation method with 95% ethanol, followed by steam evaporation. Exactly 250 g of dry
powder was percolated to get a net yield of 30.6 g of concentrated extract (12.24%). Male Wistar rats (200-300g total weight)
were anesthetized with sodium pentobarbital (45mg/kg, i.p.). Polyethylene catheters were inserted into the lower abdominal aorta
and into the inferior vena cava, for blood pressure measurements and administration of drugs, respectively. After a 24h period,
experiments were performed in conscious unrestrained rats. The results are presented as mean standard error of the mean. The
study was approved by the ethics committee of the Federal University of Alagoas (010151/2008-82). The EESCF (0.5; 1; 5; 10;
20 and 30 mg.kg-1 i.v., randomly, n=5) elicited immediate and dose-independent decreases in mean arterial pressure (MAP) (15.1 1.4%; -15.8 1.1%; -14.9 1.9%; -13.0 0.9%; -11,2 2.4%; -13,0 2.2%, respectively) with reduction in heart rate
(HR) (-5.7 1.0%; -5.3 1.3%; -5.6 0.5%; -14.2 1.5%; -8.3 1.5%; -9.6 2.4%, respectively). The hypotensive effect of
EESCF was unaltered by Methylatropine (2 mg/Kg; 30 min; i.v., n=5), however significantly reduced the bradycardia in higher
doses (10 mg/kg, only). After i.v. pretreatment of 20 min with the nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl
Esther L-NAME (20 mg/Kg; i.v., n=5), the hypotensive responses to EESCF were significantly attenuated (0.5; 5 and 10 mg/Kg)
nevertheless, bradycardia were abolished. Hypotensive responses to EESCF were significantly attenuated after i.v. pretreatment
with indomethacin for 20 min (3 mg/Kg; i.v., n=5) and cause an antagonistic effect over bradycardia. Pretreatment with i.v.
hexamethonium for 30 min (30 mg/kg; i. v., n=5) significantly reduced the hipotensive effect of EESCF (0.5 and 10 mg/kg) and
bradycardia (0.5; 5; 10 and 20 mg/kg).
Conclusions:
These results suggest that the hypotensive actions of the EESCF is probably due to a peripheral vasodilation, at least partly
secondary to the release of endothelium-derived products and of the bradycardia appears mainly dependent upon the participation
of ganglion pathway.
Keywords: HYPOTENSIVE , SYZIGIUM CUMINI , IN VIVO
Financial Support: FAPEAL, CAPES and CNPq
Resumo:26-162
INFLUENCE OF GLYCYRRHIZA URALENSIS EXTRACT ON THE PLASMID TOPOLOGY AND ON THE
STANNOUS CHLORIDE EFFECTS ON THE DNA OF PLASMIDS.
Santos, R. R. M. ; Pereira, M. O. ; Carmo, F. S. ; Jost, R. T. ; Almeida, D. S. ; Frederico, E. H. F. F. ;

Dantas, M. P. ; Santos, P. T. S. ; Filho, S. D. S. ; Bernardo-filho, M.
Departamento de Biofsica e Biometria, IBRAG/UERJ
Objectives:
Glycyrrhiza uralensis root (Alcauz) is considered one of the oldest and most widely used herbal drugs around the world.
Stannous chloride (SnCl2) is a powerful reducing agent used for different purposes and presents genotoxic effects. The objective
of this work was to investigate the effect of an aqueous G. uralensis extract (GU) on the plasmid DNA topology and on the
effects of the stannous chloride on DNA plasmid. Some physical characteristics, as visible absorbance spectrum, pH, and
refractive index of GU extract were also determined.

In the preparation of GU, 50 mL of boiled saline (0.9% NaCl) solution was added to 2 g of the extract, in a form of commercial
concentrated granules (E fong, lote number 08055431, validity 04/2013, China) and homogenized. The preparation was
centrifuged (2.000 rpm, 15 min), and the supernatant was considered to have 40 mg/mL extract and was denominated as 100%.
After that, dilutions (GU 20 mg/mL; GU 10 mg/mL; GU 5 mg/mL; GU 2.5 mg/mL) were prepared in saline, and the pH and
refractive index for each extract concentration was measured. The spectrophotometry of GU was perfomed with extract
concentration of 10 mg/mL. All measures were done in three concurrent preparations of GU extract, and the data are reported as
average and standard deviation. The pH (5.350.02), the refractive index (1.30.06), and the absorbance spectrum of the extract
(GU 10 mg/mL) were used as markers of the reproducibility of the conditions of the extract. Data showed an absorbance peak at
490 nm (0.5980.028) that was pH and refractive index concentration dependent. Plasmid DNA samples were incubated with GU
(40 mg/mL, 10 mg/mL, 2.5 mg/mL) in the presence or absence of SnCl2 (200 mg/mL) during 40 minutes. As controls plasmid
incubated only with SnCl2 or with saline (0.9% NaCl) were used. After that, 0.8% agarose gel electrophoresis (7V/cm) was
performed, the gel was stained with ethidium bromide and the plasmid topological forms (bands) were visualized and
semiquantied data are reported as percentage of supercoiled form (%SC). The results in %SC form were (XSD): 1000
(saline); 75.903.96 (GU 40 mg/mL); 1000 (GU 10 mg/mL); 1000 (GU 2.5 mg/mL); 54.24.81 (SnCl2); 72.358.84 (GU 40
mg/mL + SnCl2); 61.2511.67 (GU 10 mg/mL + SnCl2); 76.954.88 (GU 2.5 mg/mL + SnCl2).
Conclusions:
The results obtained in this work could indicate a dose-dependent genotoxic effect of GU extract and a protective action against
SnCl2 effects on the plasmid DNA.
Keywords: Glycyrrhiza uralensis, plasmid DNA, stannous chloride, genotoxic effect
Financial Support: CNPq, FAPERJ and UERJ.
Resumo:26-163
ANTIPLATELET, ANTITHROMBOTIC AND FIBRINOLYTIC ACTIVITIES OF CAMPOMANESIA
XANTHOCARPA
Klafke, J. Z. 1,3; Silva, M. A. 1; Rossato, M. F. 1; Trevisan, G. 1; Walker, C. I. B. 1; Leal, C. A. M. 1; Borges,

D. O. 3; Duarte, M. M. M. F. 2; Viecili, P. R. N. 3; Ferreira, J. 1
1
Programa de Ps-Graduao em Bioqumica Toxicolgica, UFSM
2
Departamento de Cincias da Sade, ULBRA
3
Grupo Multidisciplinar de Sade, UNICRUZ
Objectives:
One of the most recent studies demonstrated that the Campomanesia xanthocarpa produced an effect parallel with the mechanism
of oral hypolipemiants (J. Ethnopharmacol. 127; 299, 2010). Since hypolipemiants also exert antithrombotic effects, and although
C. xanthocarpa has reduced the blood cholesterol levels in hypercholesterolemic patients, until now no information has been
available about the antithrombotic effect of C. xanthocarpa. Thus, the major aim of the present work was to investigate the effects
of C. xanthocarpa extract on antiplatelet, antithrombotic and fibrinolytic activities in mice.
Male Swiss mice (30-40 g, n= 8-10) were treated orally for 5 days with 10, 30 or 100 mg/Kg of C. xanthocarpa extract (CXE) or
100 mg/Kg of acetylsalicylic acid (ASA) and at the end of the treatment period animals were challenged for bleeding, acute
thromboembolism and ulcerogenic activity. In addition, we have assessed the prothrombin time (PT) and activated partial
thromboplastin time (aPTT) after oral administration. In in vitro assays, antiplatelet effects of CXE were evaluated on ADPinduced platelet aggregation, and fibrinolytic activity of the extract was observed by artificial blood clot degradation. Moreover,
platelet citotoxity of the extract was also determined by the LDH assay. The mean amount of blood lost from control animals 2
min after tail transaction was 5.83.1 L. Mice treated with antithrombotic doses CXE, 30 and 100 mg/Kg/day, showed
significant difference from control, losing 24.36.6 and 43.414.5 L of blood, respectively. ASA treated animals lost 51.912.9
L of blood, showing a significant difference when compared to the control group. Moreover, CXE had significant
antithrombotic activity when administered at 100 mg/Kg/day since it prevented paralysis in 80% compared with control. ASA
showed no significant difference compared to the control to reduce paralysis. None of the doses of the extract were capable of
inducing ulcerogenic activity, while aspirin (100 mg/Kg/ day) induced the formation of gastric lesions [the medians (25-75
percentiles) with lesion scores of 0 (0-1); 4 (34); 0 (01), 1 (0-1) and 2 (12) for vehicle; aspirin; 10 mg/Kg, 30 mg/Kg and 100
mg/Kg/day of CXE, respectively. Besides, in the CXE -treated groups, the aPPT increased significantly to 32.300.91 sec and
33.130.63 sec, at the doses of 30 and 100 mg/Kg, respectively, although remaining in normal limits when compared to the
vehicle group. There were no significant increases of PT when compared to the vehicle group. In in vitro assays, CXE showed a
concentration-dependent inhibition of ADP-induced platelet aggregation (Imax= 365% for the concentration of 1000 g/mL;
EC50= 35 (15 84) g/mL) compared with the control group. In the cytotoxicity parameter, no concentration tested of CXE
induced LDH release, while the positive control, SDS 10%, significantly increased the LDH release. Moreover, in the evaluation
of the fibrinolytic effect, blood clot degradation was observed in all the test tubes of CXE or streptokinase (STK), with an EC50
value of 21 (5 87) g/mL and 24 (9 64) g/mL, and an Emax of 569% and 6014%, respectively, for the concentration of
100 g/mL of both.
Conclusions:
CXE showed antiplatelet, antithrombotic and fibrinolytic activities in mice. The antithrombotic activity of CXE derived probably
from antiplatelet aggregation and fibrinolytic activities.
Keywords: Bleeding, Medicinal plant, Myrtaceae, Platelet, Thrombosis
Financial Support: CAPES; CNPq; CCNE/UFSM.
Resumo:26-164
INVESTIGATION OF IN VITRO ANTICHOLINESTERASIC AND ANTIOXIDANT EFFECTS OF CARYOCAR
BRASILIENSIS CAMB.
Rodrigues, A. G. ; Silva, B. G. D. ; Oliveira, L. M. D. ; Nascimento, M. V. M. ; Costa, E. A. ; Ghedini, P.

C.
Departamento Fisiologia e Farmacologia/ICB, UFG
Objectives:
Alzheimer's disease (AD) is a progressive neurodegenerative pathology with severe economic and social impact. Considering the
presence of central cholinergic disfunction in the AD and that the free radical-induced oxidative damage may play a role in the
pathogenesis of this disease, the development of drugs with anticholinesterasic and antioxidant effects can represent compounds
more efficient for AD treatment. The natural products are potential candidates in the research of new drugs. Thus, the aim of this
work was to investigate the anticholinesterasic and antioxidant effects of Caryocar brasiliensis, a plant of Brazilian Cerrado
known as pequi.

The crude hidroalcoholic extract of leaves of C. brasiliensis (CHE) was used in this study. The acetylcholinesterase enzyme was
obtained of mice brain in phosphate buffer pH 8.0. 10 L of DTNB 0.01 M, 8 L of ASCh 0.075 M, 1.200 L of enzyme (20 g
protein/mL) and CHE at final concentrations of 1, 3, 10, 30, 100, 300, and 1000 g/mL was added to 1.5 mL cuvette and the
reaction was monitored for 10 min at 412 nm. Inhibitory activity was calculated considering the percentage of enzyme activity of
sample (CHE) in relation to the control (buffer instead of CHE). Extract concentration providing 50% inhibition (IC50) was
obtained by plotting the percentage inhibition against extract concentration. The neostigmine was used as the positive control.
The DPPH assay was used for the antioxidant activity study. 1 mL of CHE at final concentrations of 0.1, 0.3, 1, 10, 30, 100, and
300 g/mL was added to 0.4 mL of DPPH 3 mM in ethanol. The absorption was measured at 517 nm against a corresponding
blank. The ability to scavenge the DPPH radical was calculated considering the percentage of absorbance of sample (DPPH plus
CHE) in relation to the control (DPPH plus ethanol). The extract concentration providing 50% inhibition was obtained by plotting
the percentage inhibition against extract concentration. The quercetin was used as the positive control. All determinations were
done in triplicate, and the IC50 values are represented by geometric mean and 95% confidence intervals (CI). The CHE inhibited
the acetylcholinesterase enzyme with IC50 of 527 g/mL (CI: 473-602 g/mL). The IC50 neostigmine was 12 ng/mL (CI: 3-55
ng/mL). The IC50 of the extract that is able to scavenge half of the DPPH was 2.7 g/mL(CI = 1.5- 5 g/mL). The IC50
quercetin was 256 ng/mL (CI = 172-381 ng/mL).
Conclusions:
The results suggested that the Caryocar brasiliensis presents better antioxidant activity than anticholinesterasic effect. Further
studies will be conducted to elucidate the effect in vivo of this plant.
Keywords: anticholinesterasic, antioxidant, Caryocar brasiliensis
Financial Support: FAPEG
Resumo:26-165
EVALUATION OF PIPER REGNELLII CRUDE EXTRACT INFLUENCE ON HORMONE-DEPENDENT CANCER
CELL LINE OVCAR-3
Longato, G. B. 1; Fiorito, G. F. 1; Monteiro, K. M. 1; Ruiz, A. L. T. G. 1; Foglio, M. A. 2; Carvalho, J. E. 1

2
Diviso de Fitoqumica, CPQBA/UNICAMP
1
Diviso de Farmacologia e Toxicologia, CPQBA/UNICAMP
Objectives:
Ovarian carcinoma is one of the most aggressive gynecological diseases and generally diagnosed at advanced stages. The search
for new therapeutic modalities in ovarian cancer, including ways to increase the effectiveness of chemotherapy, has been target of
several studies. Previous studies reported by our research group revealed that in vitro antiproliferative activity of P. regnellii
(Miq). C. DC. var. regnellii dichloromethane crude extract (DCE) was potent and selective for ovary (OVCAR-3) cancer cell
line, with TGI (Total Growth Inhibition) value of 12.05 g/mL. This study aimed the evaluation of Piper regnellii DCE influence
on human ovarian OVCAR-3 cell line proliferation through E-screen assay, that was developed to assess the estrogenicity of
samples.
DCE was evaluated in a concentration range of 25, 2.5 and 0.25 g/mL, in the presence or absence of estradiol (10-9 M).
Anastrozole was used as positive control (400, 40 and 4M). After 144h of treatment, hormonal activity was determined by
colorimetric sulphorodamine B (SRB) assay and concentrationresponse curves were plotted. It is known that estrogen is a
potential factor of ovarian carcinogenesis, acting via two nuclear receptors (alpha and beta), but the cellular signal pathways
involved are not completely clear. When administered in association with DCE at the major dose, the estrogen did not interfere in
DCE antiproliferative activity, but at dose of 2.5 g/mL this association increased the antiproliferative activity of the crude
extract. The cell growth percentage was 10.27 in the presence and 20.06 in the absence of estrogen. The estrogen also did not
interfere in cells treated with 0.25 g/mL of DCE.
Conclusions:
These results suggest that DCE activity on this ovarian cancer cell line at dose of 2.5 g/mL was improved when associated with
estradiol, promoting a synergism response. Further in vitro and in vivo studies need to be undertaken to evaluate this contribution
in hormone-dependent ovary cancer treatment.
Keywords: anticancer activity, estrogen-receptor, hormone-dependent cancer cell line, Piper regnellii, OVCAR-3
Financial Support: FAPESP, CNPq, Capes
Resumo:26-166
EVALUATION OF CYTOTOXIC VERNONIA CROTONOIDES SUBFRACTIONS IN NON-TUMOR CELLS AND
TUMORS (U87 AND K562).
Castro, E. D. S. ; Burth, P. ; Lobo, J. F. R; Arruda, L. P. ; Fernandes, C. P. ; Rocha, L. M. ; Amorim, L. M.

F.
GCM- Instituto de Biologia, UFF
Objectives:
Evaluation of cell cytotoxic activity present in seven column elutions from a dichloromethane extract of Vernonia plant leaves in
tumor cell lines (U87 and K562) and in a non-tumor cell (Vero).
The dichloromethane extract was passed through a silica column and eluted with different volumes (1.0 and 0.5 L) of the
solvents: hexane, dichloromethane, dichloromethane/acetate and acetate. One lactone was isolated from the
dichloromethane/acetate extract. Cells were maintained in DMEM (U87 and VERO) or RPMI (K562) medium with 10% fetal
bovine serum inactivated at 560C, 100 U/ml penicillin and 100 U/ml streptomycin at 370C in humid atmosphere containing 5%
CO2. A total of 3x104 cells/ well were plated in 96-well plates and, on the next day, cells were incubated with dried column
dichloromethane and the isolated lactone previously dissolved in DMSO at different concentrations (100, 50, 25, 12.5, 25.6,
ug/ml). K562 cells were plated and treated with column eluates e and lactone in this same day. After a 24h incubation,
cytotoxicity was assessed by MTT reduction and quantified by spectrophotometry at 545nm. Results of three different
experiments in triplicate (minimum) were statistically analyzed by Graph-Prism 5 program. Results: Leaf elutions of Vernonia
were cytotoxic to tumor cell lines tested. IC50 values are expressed in the table below. Elutions from DCM Extract; Cell line
IC50 (g/ml) Vero; K562; U87 01 - Hexane (1,0 L); 73,8; 20,9; 21,1 02 - Hexane (0,5 L); 69,12; 0,97; 26,83 03 Dichloromethane (1,0 L); 18,44; 0,06; 11,17 04 - Dichloromethane/acetate 4:6 (0,5 L); 32,02; 0,03; 8,02 05 - Acetate (1,0 L);
21,14; 2,26; 0,34 Isolated Lactone; 6,45; 0,01; 4,15
Conclusions:
Preliminary cytotoxicity tests showed that eluates 02, 03, 04, 05 and lactone were more cytotoxic for K562 than for U87 cell
lines. In U87 line. the highest cytotoxicity was observed with eluate 05 and with the lactone fraction. Vero cells were more
sensitive to lactone.
Keywords: Vernonia, MTT, cytotoxicity, K562, U87
Financial Support: FAPERJ, Proppi-UFF, CAPES, REUNI
Resumo:26-167
PURIFICATION AND PARTIAL BIOCHEMICAL CHARACTERIZATION OF P6S1: A METALLOPROTEASE
FROM BOTHROPS MOOJENI VENOM.
Marinho, A. L. Z. 1,3; Queiroz, M. R. 2,3,1; Morais, N. C. G. 1,3; Sousa, B. B. 1,3; Pereira, D. F. C1; Silva, T.
K. A. 1; Oliveira, F. 1,3
1
Universidade Federal de Uberlndia, UFU
2
3
Instituto Nacional de Cincia e Tecnologia, INCT (N-Biofar)
Objectives:
The aims of this study were to purify and partially characterize a protease named P6S1 from Bothrops moojeni venom.
P6S1 was obtained after two chromatographic steps, ion exchange on DEAE-Sephacel and molecular exclusion on Sephacryl
S300. The enzyme was purified to homogeneity as judged by its migration profile in SDSPAGE stained with coomassie blue,
and showed a molecular mass of about 60 kDa. Its proteolytic activity was assayed using bovine fibrinogen as substrate at
different times (5, 15, 30, 60 and 90min), temperatures (30, 40, 50, 60, 70, 80 e 90C) and pHs (4-11). P6S1 showed high
proteolytic activity on bovine fibrinogen as substrate and it was devoid coagulant activities on bovine plasma. This enzyme
cleaves the A&alpha-chain of fibrinogen first, followed by the B&beta-chain, and shows no effects on the gama-chain. The
fibrinogenolytic activity of P6S1 was abolished after incubation with EDTA (ethylenediamine tetra-acetic acid) and 1,10phenantroline. Aprotinin and benzamidine, specific serine protease inhibitors, had no effect on P6S1 activity. Beside this, P6S1
maintained its fibrinogenolytic activity on pH from 5.0 to 11.0 and temperature from 30-40C.
Conclusions:
We have purified a protease, P6S1, with fibrinogenolytic activities, from Bothrops moojeni venom. The properties of P6S1
suggests that this enzyme is a &alpha-fibrinogease and zinc-dependent metalloprotease. Its pH and temperature optimum of
proteolysis of bovine fibrinogen was about 8 e 30C respectively.
Keywords: Bothrops moojeni , metalloprotease, snake venom
Resumo:26-168
EVALUATION OF THE TOXICITY OF THE METHANOLIC EXTRACT FROM LEAVES OF MAYTENUS
ERYTHROXYLON REISSEK (CELASTRACEAE)
Almeida, C. L. F. ; Lima, G. R. M. ; Montenegro, C. A. ; Leite, T. J. A. ; Viana, W. P. ; Tavares, J. F. ;

Castello-branco, M. V. S. ; Batista, L. M.
LTF/DCF/CCS/UNIVERSIDADE FEDERAL DA PARABA, UFPB
Objectives:
Currently there are approximately 80 species of Maytenus distributed throughout the Brazilian territory. The Maytenus genus is
characterized chemically by the presence of secondary metabolites such as flavonoids, triterpenes, alkaloids and tannins. Given
the importance of chemistry and pharmacology of the genus and the lack of studies on the species Maytenus erythroxylon, this
study proposes to investigate the toxicity of methanolic extract from leaves of M. eryhtroxylon (Me-MetOH).
To evaluate the toxicity of Me-MetOH was used the brine shrimp (Artemia salina) lethality test. 25 mg of eggs of A. saline were
incubated in seawater (pH 8-9/29C) with artificial light for 24 hours to obtain occlusion of cysts and larvae. The extract was
diluted in seawater (1-10mg/mL) and then was added 5mL of different concentrations of Me-MetOH in tubes containing 13-15
nauplii. The control group was prepared with the solvent and A. saline. The set was incubated in the presence of artificial light
(24 h) and then the surviving larvae were counted to determine the LC50 (Phytomed. 8:395, 2001). In the study of acute oral
toxicity, single dose of 2000 mg/kg was administered in groups of Swiss albino mice males and females (n= 6-10, weight: 2535g) after 12 h. The animals that received vehicle (saline 0.9%) served as controls. After treatment, the parameters of behavior
were observed for 30, 60, 90, 120, 180, 240 minutes, 24, 48 and 72 h (Rev. Bras. Sci. Farm. 80:72, 1999). Food and water
consumption were evaluated in both sexes, within 14 days. At the end of the period the number of survivors was recorded to
determine DL50. Macroscopic changes in organs of mice were evaluated. The results were expressed as meanstandard
deviation. These data were analyzed by test t of Student. The experimental protocols were approved by the Institutional
Committee for Ethics in Animal Research of LTF/UFPB. The test of lethality of brine shrimp is used for allow an initial
toxicological analysis. This test showed that the Me-MetOH has bioactivity suggesting the presence of important bioactive
substances in it, because the LC50 (827,1 g/mL) was less than 1000 mg/mL. The acute oral toxicity consists of a preliminary
assessment of the toxic properties of a substance test. A single dose of Me-MetOH (2000 mg/kg) not induce changes in behavior.
During the 14 days of observation there were no deaths and found no increase in body weight of animals at the start of the
protocol. Me-MetOH not induced macroscopic changes in the organs of mice. It was observed that the extract didnt cause
significant change in water consumption of females, whereas there was significant decrease in water intake of treated males (MeMetOH:79,059,370***;control:101,716,42). In relation to feed intake was observed that Me-MetOH not promote any change
in the treated groups.
Conclusions:
Me-MetOH presented bioactive front A. salina and despite the observed change is not possible to infer toxicity to the extract.
Thus it is possible to suggest that Me-MetOH single dose of 2000 mg/kg showed low toxicity in the evaluated conditions.
Keywords: Artemia salina, extract, Maytenus erytrhoxylon, natural products, toxicology
Financial Support: CAPES/CNPq/LTF/UFPB
Resumo:26-169
EVALUATION OF THE GASTROPROTECTIVE ACTIVITY OF COMBRETUM DUARTEANUM CAMBESS

(COMBRETACEAE) LEAVES: ROLE OF ENDOGENOUS SULPHYDRYLS AND NITRIC OXIDE
Lima, G. R. M. ; Montenegro, C. A. ; Leite, T. J. A. ; Almeida, C. L. F. ; Cabral, A. G. S. ; Gomes, I. F. ;

Tavares, J. F. ; Batista, L. M.
Objectives:
Combretum duarteanum Cambess (Cd), unique species of South America, registered in Bolivia, Paraguay and Brazil. Its
occurrence in Paraiba is restricted to the Caatinga, usually found in rocky outcrops. It is popularly known as mofumbo or
cipiba. Based on the pharmacological effects, this work aims to evaluate the gastroprotective activity and the likely
mechanisms of related actions of the ethanolic extract (Cd-EEtOH) and the hexane phase (Cd-FaHex) obtained from the leaves of
C. duarteanum Cambess.
The Cd-EEtOH and Cd-FaHex were evaluated for the ability to protect the gastric mucosa against injuries caused by pylorus
ligation induced ulcer model in rats and participation of gastric wall mucus, sulfhydryls (SH) groups and NO (nitric oxide) in the
Cd gastroprotective action. Gastric secretion studies were done by pyloric ligation experiment. Male Wistar rats (180250 g)
were treated with Cd-EEtOH (250 mg/Kg), Cd-FaHex (250 mg/Kg), carbenoxolone (100 or 200 mg/kg), cimetidine (100 mg/kg)
or the tween 80 solution 12% (10 mg/kg) (n=5-7). The ulcerative lesion index (U.L.I.) is expressed in meanS.D and were
compared using ANOVA followed by Dunnetts or Tukeys test, pp>0, 05). Rats treated with Cd-FaHex did not increase free
mucus production (p>0.05). For ethanol-induced gastric lesion, pretreatment with N-ethylmaleimide (NEM) (10 mg/kg, i.p.), a
blocker of endogenous sulphydryl group, markedly increased the gastric lesions when compared to control groups. Animals
treated with Cd-EEtOH or Cd-FaHex showed a significant reduction of the U.L.I. (67%; 46% with tween 80 (12%) versus 39%;
40% with NEM). To investigate the role of endogenous NO in cytoprotection, we used the NO synthase inhibitor (L-NAME) to
access the protective effect of the Cd. Oral administration of Cd-EEtOH or Cd-FaHex to animals pretreated with L-NAME (70
mg/kg, i.p.) produced a rise in gastric damage when compared to pretreated with tween 80 (12%). Therefore, the gastroprotective
effect of Cd-EEtOH and Cd-FaHex observed by pretreatment with tween 80 (12%) (67,1710,72; 117,549,59) was significantly
reduced (p
Conclusions:
These results suggest that the Cd-EEtOH and Cd-FaHex displays gastroprotective effect, but did not exhibited antisecretory
activity. The observed gastroprotectives effects probably involve the participation of NO as well as an increase in endogenous SH
compounds, which are related to the protective mechanisms of the gastrointestinal mucosa against aggressive factors.
Keywords: Combretum duarteanum Cambess, gastric ulcer, gastroprotective activity, Sulfhydryl compounds, Nitric oxide
Financial Support: CAPES/CNPq/LTF/UFPB.
Resumo:26-170
PHARMACOLOGICAL STUDY OF P-MENTHANE ESTERS: STRUCTURE-ACTIVITY RELATIONSHIP
Andrade, L. N. ; Matos, R. M. ; Santos, D. S. ; Lima, T. C. ; Batista, J. S. ; de Sousa, D. P.

Departamento de Fisiologia/ Universidade Federal de Sergipe, UFS
Objectives:
Several essential oils have exhibited spasmolytic effect in various types of smooth muscles. This effect has been attributed to
monoterpenes, major compounds and of varied structural diversity. Therefore, the main of the present work was to evaluate the
structure-spasmolytic activity relationship of ten p-menthane esters derived from terpenes in isolated guinea-pig ileum as well as
to investigate the mechanism of action of the more potent ester.
The acetates of menthyl (1), perillyl (2), -terpinyl (3), p-menth-1-en-9-yl (4), thymyl (5), isopulegyl (6), 4-terpinyl (7), carveyl
(8), carvacryl (9) e neo-isopulegyl (10) were obtained from acetilation of p-menthane monoterpenes. The relaxant activity of
these compounds was examined in guinea pig isolated ileum pre-contracted by bethanechol (30 M). The next step consisted in
the evaluation of the mechanism of relaxant action of 4-terpinyl acetate (10 M), one of the more potent compounds. To this
purpose, were evaluated the involvement of voltage-dependent calcium channels (curve to calcium were obtained in absence and
in presence of nifedipine 10 M and of 7), and of potassium channels (relaxation induced by 7 were obtained in pre-contracted
ileum by bethanechol in the absence and presence of tetraethylammonium TEA - 1 mM) in the relaxation induced by 7. In
addition, it was investigated the involvement of muscarinic receptors antagonism (concentration-response curves to bethanechol
was obtained in the absence and in the presence of 7), and histaminergic receptors antagonism (contraction induced by histamine
0.15 M was obtained in the absence and in the presence of 7) in the relaxant response induced by 7. Statistical analysis was
performed by one-way Anova with Tukeys post-test or by Student's test. The EC50 values to the relaxant activity of the
compounds 1, 2, 3, 4, 5, 6, 7, 8, 9 e 10 were 73, 220, 53, 27, 17, 16, 10, 30, 37 e 6.5 M, respectively. The calcium-induced
contraction was fully blocked by nifedipine. However, 4-terpinyl acetate did not shift the calcium curve. Moreover, pre-treatment
with TEA did not inhibit the relaxation induced by 7. Therefore, these results suggest that the relaxation induced by 7 does not
involve either voltage-dependent calcium blockage or potassium channels activation. Contraction produced by histamine also was
not inhibited by 7, which suggests that this compound do not produce histamine receptors antagonism. On the other hand, 4terpinyl acetate shifted the bethanechol curve to the right with reduction of the maximum response (bethanechol CE50 values
were 0.7 (0.4-1) M and 2 (1-4) M in the absence and the presence of 7, respectively). Thus, this result suggests that 7 produces
antagonism of muscarinic receptors.
Conclusions:
These results suggest that the relaxant activity of 4-terpinyl acetate in guinea pig isolated ileum is not involve either blockade of
voltage-dependent calcium channels, or activation of potassium channels or antagonism of histaminergic receptors. The most
likely mechanism involves muscarinic receptors antagonism.
Keywords: atividade espasmoltica, steres p-mentnicos, mecanismo de ao, relao estrutura-atividade
Financial Support: Cnpq and CAPES
Resumo:26-171
ESSENTIAL OIL FROM PTERODON EMARGINATUS SEEDS PRODUCES ANTIINFLAMMATORY AND
ANTIEDEMATOGENIC EFFECT IN MICE
Nucci, C. 1; Mazzardo-martins, L. 3; Stramosk, J. 2; Ludtke, D. D. 2; Santos, A. R. S. 3; Martins, D. F. 1,2,3

1
2
3
Objectives:
Pterodon emarginatus, known as sucupira branca, is a native tree widely distributed over the central region of Brazil. Although
an anti-rheumatic active compound seems to be present in their oleaginous fraction, seeds hydroalcoholic (wine) infusions are
used in folk medicine, throughout Brazil, for their anti-rheumatic, analgesic and anti- inflammatory properties. Phytochemical
studies on Pterodon have shown the presence of diterpenes and isoflavones in seed oil. Furthermore, toxicological studies have
demonstrated that oral administration of high doses of Pterodon emarginatus seed oil did not produce acute toxicity effects in
healthy mice. This study examined the effect of ethanolic extract of Pterodon emaginatus in inflammatory nociception model
induced by formalin in mice.
Committee of the Universidade do Sul de Santa Catarina (UNISUL). The ethanolic extract of the Pterodon emarginatus was
prepared with 100g of the fruit (pod) with the crushed seeds in 1 liter of alcohol. Then, placed in a dark glass to macerate for 30
days in a fresh and airy. After this time the compound was then strained and passed by the vaporization process for essential oil
extraction plant. The essential oil of Pterodon emarginatus (EOP) was diluted in saline solution (NaCl, 0.9%) and Tween (5%) to
intragastric treatment (i.g.) in mice. The control group was treated only with vehicle (saline 0.9% and Tween 5%). To examine
the antinociceptive and antiinflammatory effect of EOP we used intraplantar injection of formalin. Mice were injected with
formalin (20 l of 2.5% formalin (0.92% formaldehyde) in phosphate-buffer saline) or saline in the ventral area of the right and
left hind paw, respectively. Licking of the injected paw, recorded as nociceptive responses, were measured 0-5 min. (first phase)
and 15-30 min. (second phase) after formalin injection. The animals were treated with a solution containing the EOP (3-100
mg/kg, i.g.) 1h before the tests. Control groups were treated with vehicle (10 ml/kg. i.g.). The effect of antiedematogenic of EOP
was verificated after intraplantar injection of formalin, by measuring the difference in weight and thickness of the paws mice. To
investigate the effect of EOP on locomotor activity we used the open field test. Our results shown that a 60-minute pretreatment
with the essential oil Pterodon emarginatus (EOP) (3-100mg/kg, i.g.) significantly inhibited the inflammatory (late) phase of
formalin- induced pain (6572% inhibition) and, to a lesser extent, neurogenic (early phase, 4395% inhibition) formalininduced nociception. The mean ID50 value for the late phase was 3.63mg/kg (2.76-4.51mg/kg confidence intervals).
Furthermore, the EOP also produced antiedematogenic effect with reduction of 5211% at the dose of 100 mg/kg, i.g. on
thickness and 2316% on weight of the paw. However, the treatment of animals with the EOP in different doses (1-100mg/kg,
i.g.), did not affect the motor performance in the open-field test.
Conclusions:
We demostrated that the essential oil extracted from seeds of Pterodon emarginatus produced antinociceptive and antiinflammatory effect in the formalin model of nociception in mice.
Keywords: Inflammation, Pterodon emarginatus, Mice, Sucupira
Financial Support: UFSC, Capes, UNISUL (Artigo 170).
Resumo:26-172
A PROTEOLYTIC FRACTION FROM VASCONCELLEA CUNDINAMARCENSIS (CARICA CANDAMARCENCIS)
LATEX ALTERS HEMOSTATIC PATTERNS IN VITRO AND DISPLAYS ANTITHROMBOTIC ACTIVITY IN
VIVO.
Bilheiro, R. P. 1; Braga, A. D. 1; Filho, M. L. 1; Carvalho-tavares, J1; Salas, C. E. 1; Agero, U. 4; Carvalho,

M. G. 3; Lopes, M. T. P. 1
Depto de Farmacologia/Instituto de Cincias Biolgicas/UFMG, ICB / UFMG

2
Depto de Fisiologia / Instituto de Cincias Biolgicas, ICB / UFMG
3
Faculdade de Farmcia da UFMG, Fac de Farmcia/UFMG
4
Depto de Fsica/Instituto de Cincias Exatas, ICEx/UFMG
Objectives:
In previous studies, P1G10, a cistein proteases-rich proteolitic fraction of the latex from fruits of Vanconcellea
Cundinamarcensis, has affected various physiophatological systems, such as mitogenesis, angiogenesis, antitumor and
antimetastatic effect, gastric wound healing and cicatrization of skin injuries. In the acute toxicity study of P1G10 (50mg/kg, i.p.
or i.v), signs of hemorrhage were observed in the stomach and intestines of Wistar rats of both sexes. In this study, we assessed
the effect of P1G10 on plasma coagulation and on platelet aggregation as well as its action on a model of thrombotic obstruction.
For the coagulation assays, 50L of plasma from male Wistar rats ( 200g, 8 weeks, n=6/group) were incubated at 37oC for 5
minutes with either P1G10 (0.25, 0.50, 1.0g/L) or saline. The Activated Partial Tromboplastin Time (APTT) (Actin, DadeBehring, USA), the Prothrombin Time (PT) (Thromborel, Dade-Behring, USA) and the Thrombin Time (Biopool Thrombin,
Biopool, USA) assays were then performed in a coagulometer (BFT II, Dade-Behring, USA). In the coagulation assays, P1G10
significantly increased PT at 1.0g/L (15.8 1.0s, 1.4 fold) compared to control (10.7 0.2s), whereas a dose-dependent
significant increase in APTT was found at 0.5g/L (47.2 2.6s, 2.5 fold) and at 1.0g/L (92.4 4.8s, 4.8 fold) compared to
control (19.0 0.8s), and no increase found at 0.25g/L. Also, by the TT assay, a significant increase in the coagulation time
was found at 0.5g/L (142.5 5.7s, 2.5 fold) and at 1.0g/L (interrupted at 250s due to absence of coagulation) compared to
control (56.6 9.1s) (p
Conclusions:
The data shows that P1G10 prevented the formation of a stable thrombus in vivo and that this effect might be caused, at least
partially, by the inhibition of both plasma coagulation and platelet aggregation
Keywords: Carica candamarcensis, Vasconcellea cundinamarcencis, Hemostasis, Thrombosis, Intravital
Financial Support: FAPEMIG CAPES CNPq
Resumo:26-173
EFFECT OF THE GREEN TEA-DERIVED POLYPHENOL EPIGALLOCATECHIN GALLATE IN THE 6HYDROXYDOPAMINE RAT MODEL OF PARKINSONS DISEASE.
Batassini, C. ; Abib, R. T. ; Dutra, M. F. ; Borsoi, M. ; Lazzaretti, C. ; Silvestrin, R. B. ; Sorrentino, J. M. ;

Gonalves, C. A. ; Mello E Souza, T. ; Gottfried, C.
Objectives:
Parkinsons Disease (PD) is a neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia
nigra (SN) and consequent depletion of striatal dopamine. Available treatments are based on dopaminomimetic strategies but they
do not prevent disease progression, strengthening the importance of new therapies research. Green tea consumption has shown
numerous benefits to human health, and these effects have been attributed to catechins, especially epicatechin gallate (ECG) and
epigallocatechin gallate (EGCG). In this study, our aim was to evaluate the possible neuroprotective action of EGCG in a rat
model of PD induced by 6-hydroxydopamine (6-OHDA).
Male Wistar rats (110-150 days old; 250-400 g) received a single intraperitoneal (i.p).injection/day for seven days, containing 10
mg/Kg of EGCG or saline (NaCl 0.9%). In the fourth day of the treatment, they received three stereotaxic infusions of 6-OHDA
(6 microlitros; 21 microgramas) or vehicle (NaCl 0.9% + ascorbic acid 0.2%, sham) into the right striatum. The animals were
randomly assigned into four groups: (a) saline+sham; (b) saline+6-OHDA; (c) EGCG+sham; (d) EGCG+6-OHDA. The
assessment of rotational behavior induced by methylphenidate (MF) (40 mg/kg, 4 mg/mL, i.p.) took place 30 days after
stereotaxic surgery. Sixty days after stereotaxic surgery, an anti-tyrosine hydroxylase (TH) immunohistochemistry was performed
in order to identify TH-positive cells in the SN pars compacta (SNpc). The images obtained were analyzed using the Scion Image
for Windows program by pairing the amount of nigral TH-positive neurons in the SNpc from both hemispheres, whereas the
value of contralateral hemisphere to the 6-OHDA administration site was normalized to 100%. All protocols were approved by
local Research Ethical Committee (project number 19553). The statistical analysis was performed using the two-way ANOVA
test followed by Duncan post-hoc test. P< 0.05 indicated statistical difference. There was an increase in the number of ipsilateral
rotations in animals lesioned with 6-OHDA [lesion effect factor; F(3,16)=10.44; p=0.002], confirming the induction of the model.
However, there was no significant difference in the number of rotations between groups [treatment effect factor; F(3,18)=1.77;
p=0.19] (n = 13, 9, 10, 11 in the groups saline+sham, EGCG+sham, saline+6-OHDA and EGCG+6-OHDA, respectively).
Animals treated with EGCG showed a smaller reduction in immunocontent of TH when compared with animals treated with
saline [treatment effect factor; F(1,10)=11; p=0.007]. (n=2, 4, 4, 4 in the groups saline+sham, EGCG+sham, saline+6-OHDA and
EGCG+6-OHDA, respectively).
Conclusions:
Our preliminary results suggest that EGCG may exert a neuroprotective effect in the 6-OHDA PD model, since we found a
smaller decrease in the number of TH-positive cells in EGCG-treated animals when compared to the control animals. However,
EGCG has not shown a beneficial effect on motor behavior.
Keywords: GREEN TEA, EPIGALLOCATECHIN GALLATE , PARKINSONS DISEASE, 6-HYDROXYDOPAMINE
Financial Support: CNPq, CAPES, FAPERGS
Resumo:26-174
INFLUENCE OF THE TEMPERATURE ON LEAVES OF I. SUFFRUTICOSA TO OBTAIN BIS INDOL ALKALOIDS
Vieira, J. R. C. ; Lima, I. R. D. ; Silva, I. B. D. ; Nascimento, A. F. L. ; Leite, S. P.

Histologia e Embriologia, UFPE
Objectives:
Indigofera suffruticosa Mill (Fabaceae), popularly known as anil, occurs in abundance in Northeastern Brazil and has intensive
popular use in the treatment of infections, inflammation and other processes, without reports of harmful effects to humans. The
storage temperatures of plants are important factors to obtain chemical natural products. To extract bis indol alkaloids (indigo and
indirubin) efficiently, this study was developed to determine the best storage environmental temperature of leaves of I.
suffruticosa.
The obtainment of indigo and indirubin was performed using 300 g of leaves divided into two equal parts (150g) stored at 5C
and room temperature for 4 days. The plant material was submitted to aqueous extraction to investigate the presence of bands of
indigo and indirubin. The bands were analyzed by Thin Layer Chromatography (TLC) using as mobile phase system: ethyl
acetate 100 v.v., formic acetate 2 v.v., acetic acid 2 v.v., water 2v.v. The silica gel plates were revealed with vanillin
hydrochloride. Leaves of I. suffruticosa stored at 5C revealed the presence of indigo and indirubin with retention factor of 0.75
and 0.50 respectivily, and at room temperature absence of the derivative indigoids.
Conclusions:
The TLC showed the presence of bands of indigo and indirubin from the aqueous extract of leaves stored at 5C and absence of
bands at room temperature. This condition determines the influence of temperature on the bis indol alkaloids of I. suffruticosa.
Keywords: Indigofera suffruticosa, Bis indol alkaloids, Indigo, Indirubin
Resumo:26-175
TEPHROSIA SINAPOU EXTRACT REDUCES INFLAMMATORY LEUKOCYTE RECRUITMENT IN MICE:
EFFECT ON OXIDATIVE STRESS, NITRIC OXIDE AND CYTOKINE PRODUCTION
Martinez, R. M. 1; Zarpelon, A. C. 2; Zimermann, V. V. M. 1; Andrei, C. C. 3; Fonseca, M. J. V. 4; Vicentini,

F. T. M. D. C. 4; Moreira, I. C. 5; Baracat, M. M. 1; Georgetti, S. R. 1; Verri, W. A. 2; Casagrande, R. 1
1
Cincias Farmacuticas/Universidade Estadual de Londrina, CCS/UEL
2
Cincias Patolgicas/Universidade Estadual de Londrina, CCB/UEL
3
Qumica/Universidade Estadual de Londrina, CCE/UEL
4
Cincias Farmacuticas/ Faculdade de Cincias Farmacuticas , FCFRP/USP
5
Universidade Tecnolgica Federal do Paran, UTFPR
Objectives:
Tephrosia sinapou is a tropical plant and is known as a source of flavonoids. Flavonoids are phenolic antioxidant compounds,
and this activity might explain their other effects on modulation of inflammation, nevertheless, there is no study on the T. sinapou
effects on inflammation. Therefore, it was investigated the in vitro effect of T. sinapou ethyl acetate extract on oxidative
stress/antioxidant mechanism, and the in vivo effect on inflammatory leukocyte recruitment, and the participation of nitric oxide
and cytokines in the extract anti-inflammatory mechanism in mice.
The antioxidant activity of T. sinapou ethyl acetate extract was determined by the DPPH, ABTS scavenging, iron-chelating, irondependent and iron-independent lipid peroxidation activity tests. The T. sinapou extract concentration-dependently scavenged the
negatively charged DPPH radical (IC50 = 142 micrograms/ml) and the positively charged ABTS radical (IC50 = 54.51
micrograms/ml). The extract also concentration-dependently inhibited the iron-independent and iron-dependent lipid peroxidation
and iron chelation assays with IC50 of 8.53 micrograms/ml, 20.10 micrograms/ml and 1.34 mg/ml, respectively. Male Swiss
mice (20-25g; protocols approved by the Ethics Comitee of UEL) were treated with vehicle (20% tween 80 in saline,
intraperitoneally [i.p.]), T. sinapou extract (10-100 mg/kg, i.p.) or prednisolone (400 micromolar/kg, p.o.) 30 min before
carrageenin (500 micrograms/mice, i.p.) stimulus. There was dose-dependent inhibition of carrageenin-induced total leukocytes
and neutrophil recruitment by T. sinapou extract (78 and 88 %, respectively). In another set, mice were treated with L-NAME
(nitric oxide synthase inhibitor; 10-100 mg/kg, s.c.) 30 min before T. sinapou (100 mg/kg, i.p.) treatment, which reverted the
extract inhibition of carrageenin-induced total leukocytes (71%) and neutrophil (81%) recruitment. The extract also inhibited
zymosan (100 micrograms/mice, i.p.)-, glycogen (500 microliters of 5% glycogen, i.p.)-, and lipopolysaccharide (LPS, 100
nanograms/mice, i.p.)-induced total leukocytes and neutrophil recruitment by 61 and 82%; 62 and 70%, and 73 and 82%,
respectively. The carrageenin-induced TNFalfa and IL-1beta production in the peritoneal cavity was also inhibited by T. sinapou
by 94% and 74%, respectively.
Conclusions:
This study provides the first pre-clinical evidence that the extract of T. sinapou presents anti-inflammatory effects by mechanisms
involving the inhibition of oxidative stress, nitric oxide-dependent inhibition of leukocyte recruitment, and inhibition of cytokine
production.
Keywords: Tephrosia sinapou, Inflammation, Oxidative stress, Nitric oxide, Cytokine
Financial Support: Fundao Araucria and CNPQ
Resumo:26-176
VASORELAXANT ACTIVITY OF THE FLAVONOID NARINGENIN AND ITS METABOLITE OBTAINED BY
MICROBIAL BIOCONVERSION
Rocha, M. L. ; Penso, J. ; Martins, D. R. ; Mendanha, C. R. ; de Oliveira, V.

Faculdade de Farmcia, UFG
Objectives:
Naringenin is a flavonoid of considerable therapeutic interest, reported to modulate vascular tone and present protective
cardiovascular effects. Despite this, its low water solubility often presents a shortcoming for biological applications.
Glycosylation is useful for improving bioavailability and pharmacological properties. In this study, we aimed to glycosylate
naringenin in good yield using cell biocatalysts and compare vascular reactivity of the naringenin and its glycosylated metabolite.
Methods: For bioconversion experiments, Erlenmeyer flasks containing sterile broth were inoculated with a spore suspension
(one flask for each strain) and then incubated at 27 C and 200 rpm in a rotatory shaker. After 65h, naringenin was added to a
final concentration of 0.5 mg/mL and the flasks were maintained under the same conditions to allow bioconversion. Samples of
bioconversion broth were taken every 24 hours for HPLC analysis and determination of the reaction kinetics. After 96 hours,
mycelia were extracted with ethyl acetate and the resulting fraction was purified by flash-chromatography, using silica gel 60 and
ethyl acetate/methanol (70:30 v/v). For the experiments of the vascular reactivity, aortic rings were isolated from male Wistar rats
(180-200g) and mounted for isometric tension recording in an organ bath. Thus, the aortic rings with endothelium pre-contracted
with phenylephrine were exposed to naringenin, (0.5 to 500 M), its metabolite naringenin-7--O-glucoside (0.5 to 500 M) or
vehicle (DMSO). The same protocols were repeated in presence (20 min) of non-selective K+ channel blocker,
tetraethylammonium (TEA, 1 mM). Results: Beauveria sp. strains converted naringenin to one major metabolite. The strain with
best yield afforded 38% of the product, which was identified as naringenin-7--O-glucoside on the basis of spectroscopic
techniques including ESI-MS/MS, 1H and 13C NMR, HSQC and HMBC. Analyzing the biotransformation time-course of
naringenin with cultured B. bassiana cells, we observed that formation of 7--O-glucoside started at 24 hours. In the vascular
study, relaxation induced by naringenin and its metabolite have evoked concentration-dependent relaxation. The maximal
response to naringenin (84.1 2.2%, n=5) was approximately 37% higher (P0.05). The vehicle (DMSO) does not produced
relaxation. Interestingly, TEA reduced the vasorelaxant effect only to metabolite (from 48.0 3.3% to 26.3 3.2%, P
Conclusions:
Our results have shown that the presented biotransformation method is useful to convert naringenin to its 7--O-glucoside in good
yield. This metabolite is also able to induce vasodilation, although this relaxation is less effective. Moreover, it is interesting to
note that, the cellular mechanism of action responsible for relaxation seems to involve different pathways.
Keywords: aorta, flavonoid, naringenin, relaxation
Financial Support: CAPES/CNPq
Resumo:26-177
NEUROPROTECTIVE EFFECT OF ANACARDIC ACID IN EXPERIMENTAL MODEL OF PARKINSON'S
DISEASE
Medeiros-linard; C. F. B. 1; Pereira, R. D. C. R. 1; Sereniki, A. 1; Silva, S. N. 1; Amorim, A. F. C. 1; Neto, H.

P. D. F. 1; Trevisan, M. T. S3; Lafayette, S. S. L. 2
1
DEPARTAMENTO DE CINCIAS FARMACUTICAS, UFPE
2
3
DEPARTAMENTO DE QUMICA ORGNICA E INOGNICA, UFC
Objectives:
Research has found that the developments of neurodegenerative diseases are associated with high levels of oxidative stress,
mitochondrial alterations and apoptosis. The presence of oxidative stress in Parkinson's disease is associated with increased lipids
oxidation and the generation of reactive oxygen species, which are unstable chemical species. Anacardic acid is a phenolic
derivative with biological and pharmacological property. Lipoxygenase inhibitor, parasiticide, antibiotic, gastroprotective, anti
Helicobacter pylori, anticarcinogenic, antimutagenic and antioxidant activities have been identified; however no neuroprotective
in vivo activity has so far been reported according to a literature survey. This study aims to investigate the effect of anacardic
acid in behavioral tests of animals with subcutaneous rotenone induced Parkinson's.
Male Wistar rats, weighing 280-350g were used in these experiments, divided into five groups (n=6). They received rotenone
3mg/kg subcutaneous for 5 days and were treated orally with water or anacardic acid at doses of 25, 50 and 100mg/kg for seven
days, while the control animals received subcutaneous soybean oil and DMSO and were treated orally with water. After this
period were performed the following behavioral tests: open field test and elevated plus maze with all animals. Data are expressed
as mean s.e.m. Differences between groups were analyzed by ANOVA and Tukeys test (p
Conclusions:
Anacardic acid promoted a significant improvement of parkinsonian symptoms in behavioral tests of open field and elevated plus
maze.
Keywords: ANACARDIC ACID, OPEN FIELD TEST, PARKINSON
Resumo:26-178
ANTIOXIDANT EFFECT OF ANACARDIC ACID IN RATS WITH PARKINSON'S DISEASE
Medeiros-linard, C. F. B2; Amorim, A. F. C. 2; Sereniki, A2; Ribas, R. D. C. 2; Silva, S. N. 2; Wanderley, A.

G. 1; Lafayette, S. S. L. 1
1
2
DEPARTAMENTO DE CINCIAS FARMACUTICAS, UFPE
Objectives:
Studies had shown that oxidative stress and mitochondrial alterations are intimately involved in the cell death process by
apoptosis. This study aims to analyze the administration effects of anacardic acid in the rotenone model of Parkinsonism in rats.
Male Wistar rats, weighing 280-350g were used in these experiments divided into five groups (n=6). They received rotenone
3mg/kg subcutaneously for 5 days and were treated orally with water or anacardic acid at doses of 25, 50 and 100mg/kg for seven
days, while control animals received subcutaneously soybean oil and DMSO and were treated orally with water. After this period
the animals were anesthetized and sacrificed by decapitation, the brains removed. Striatum, cortex and substantia nigra were
dissected quickly on a cold plate, weighed and homogenized (10% w/v) in PBS with 1% addition of 0.004% BHT. Then the
homogenates were centrifuged at 10.000 rpm for 15 minutes at 4C and supernatants were frozen at -80C for later proteins
measurement and lipid peroxidation evaluation. Data are expressed as mean s.e.m. Differences between groups were analyzed
by ANOVA and Dunn's or Student-Newman-Keuls test (p
Conclusions:
Thus, our data demonstrate that Anacardic acid was able to prevent oxidation induced by rotenone.
Keywords: PARKINSON'S , ANTIOXIDANT , ANACARDIC ACID
Financial Support: FACEPE; CAPES
Resumo:26-179
ANTIMYCOBACTERIAL ACTIVITY AND INHIBITION OF NITRIC OXIDE PRODUCTION BY EXTRACT AND
FRACTIONS FROM PSYCHOTRIA NUDA
Heggdorne-arajo, M. 1; Ventura, T. L. B. 1; Gomes, M. V. S. 2; Oliveira, P. F. 2; Lassounskaia, E. 1;
Muzitano, M. F. 2
1
LBR/CBB - Universidade Estadual do Norte Fluminense, UENF
2
Farmcia - Universidade Federal do Rio de Janeiro/Maca, UFRJ/Maca
Objectives:
The Tuberculosis, infectious diseases caused by Mycobacterium tuberculosis, represents a challenge in worldwide. The treatment
involves the use antibiotics for a long period (6-9 months) leading frequently to the abandonment of treatment (Drug Discov
Today. 11: 21, 2006). The World Health Organization reported that 9.4 million of new tuberculosis cases occurred in 2008 and
1.3 million people died of this disease in the same year. Studies focusing antituberculosis activity of natural products are more
common. And the persistence of the Mycobacterium to antimycobacterial drugs highlight the necessity for search of new drugs to
tuberculosis treatment. The aim this study is measure antimycobacterial activity and anti-inflammatory evidences in order to
assist in the immunopathology of tuberculosis.
Samples of Psychotria nuda: Extract (PnE), infusion (PnI) and the following fractions in hexane (PnH), dichloromethane (PnD),
residual dichloromethane (PnRD), acetate (PnA), butanolic (PnB) and aqueous (PnAq) were evaluated for their antimycobacterial
activity using a tetrazole salt (3-[4,5-dimetiltiazol-2-il]-2,5-difenil-tetrazol MTT 5mg/mL em PBS estril). Initially, 50L of a
suspension of Mycobacterium bovis BCG in Middlebrook 7H9 medium supplemented with 0.05% Tween 80 and 10% ADC
(albumin-dextrose-catalase) were plated in a 96-well plate at 1 106 CFU/well and then was added 50L of medium culture
(negative control), rifampin (positive control) and of each sample in three concentrations. The plate was incubated at 37C and
5%CO2 for 7 days. After this period, 10L of MTT was added, and 3 hours after this 100L of lyses buffer. The plate was
incubated overnight and the reading was made using a spectrophotometer at 570 nm. The determination of the inhibition nitric
oxide production (NO) by macrophage and cytotoxicity was evaluated in the presence of Psychotria nuda samples. RAW 264.7
murine macrophages (5 104 cells/well) were stimulated by LPS (Lipopolissacardeo - Escherichia coli 055:B5) [1g/mL] and
incubated with samples in the concentration of 100, 20 and 4 g/mL by 24 h (n=3). The amount of NO in the culture supernatants
was determined by Griess test. The cytotoxicity was determined through specific release of LDH. In the evaluation of
mycobacterial growth, PnH (83.62%4.10), PnB (81.50%3.67), PnA (81.42%1.58), PnD (75.98%7.67), PnE (70.24%3.12)
and PnRD (60.54%0.41) in the concentration of 100g/mL showed inhibition while PnI and PnAq not showed significant
inhibition. Among the samples tested than not presented citotoxicity and inhibited the NO production, the PnA (55.33%4.07) at
20g/mL and PnD, PnRD, PnB as well as PnA at 100g/mL showed inhibition of nitric oxide production higher than 95% (data
from two independent experiments with similar results).
Conclusions:
In conclusion, samples of Psychotria nuda possibly have active substances against Mycobacterium bovis BCG that could be
active also against more virulent Mycobacterium species. In addition, properties as inhibition of the NO production and low
citotoxicity may contribute in the search of new drugs more promising for tuberculosis treatment.
Keywords: Mycobacterium bovis BCG, Natural products, Tuberculosis
Financial Support: CNPq, FAPERJ and UENF
Resumo:26-180
IN VITRO EVALUATION OF ANTI-NEOPLASTIC ACTIVITY OF THE FRUIT EXTRACT FROM SCHINUS
TEREBINTHIFOLIUS .
Marini, M. B. 1; Freitas, W. R. 1; Bernardes, N. R. 3,1; Martins, S. M. 4; Oliveira, D. B. D. 3; Muzitano, M. F.

4
; Kanshiro, M. M. 1
1
LBR/CBB- Universidade Estudal do Norte Fluminense, UENF
3
LTA/CCTA - Universidade Estadual do Norte Fluminense, UENF
4
Farmcia - Universidade Federal do Rio de Janeiro/Maca, UFRJ/Maca
Objectives:
In the last thirty years cancer appears as a disease of high world impact, being responsible for 13.7% of deaths registered in
Brazil. The anti-neoplastic treatment is very aggressive and causes many collateral effects that limit the effectiveness. Therefore
the search for new compounds with anti-neoplastic activity is fundamental. The objective of that work is to evaluate in vitro the
anti-proliferative activity of the methanolic extract of the fruits of Schinus terebinthifolius (pepper tree). It was also evaluated the
methanolic extract 1B fraction and apigenin, a flavonoid isolated from this fraction.
Methanolic extract, Fraction 1B and Apigenin were incubated with human leukemia cell line U-937 and cell viability was
evaluated by MTT assay after 48 hour of incubation. After the treatment period, it was possible to verify that 20g/mL the
methanolic extract of the pepper tree fruit reduced the cellular viability in 94,88,7% when compared with the controls cells. The
1B fraction reduced 991,3% of the cell viability and the flavonoid treatment reduced in 1000,5% the viability of the cells. In
order to verify the type of cell death induced by the extract and compound, cells were treated for 12, 24 and 48 hours and the
apoptosis evaluation was performed. Cells were stained with acridine orange plus ethidium bromide (1mg/mL) and apoptosis,
necrosis and normal cells were discriminated by fluorescence microscope examination. Methanolic extract at 20g/mL were able
to induce 50,0%10,3 of the apoptosis in 12 hour of incubation. In 48 hours of treatment 81,7%3,4 of the cells were dead by
apoptosis when treated with 2g/mL. The methanolic extract 1B fraction was active in the concentration of 20g/mL, promoting
39,1%0,7 of apoptosis with 48 hours of treatment. When treated with Apigenin for 48 hours 0,2 g/mL induced 37,9%2,1 of
cellular death by apoptosis and 20g/mL the extract induced death by apoptosis in 90,4%8,0. The cellular deaths for necrosis
didn't overcome 1% of the experimental cell, and the percentage of apoptosis in the controls groups were less than 2,5%. In
addition, the comet assay was performed to corroborate the cell death by apoptosis.
Conclusions:
Our results shows that Apigenin isolated from the methanolic extracts of pepper tree fruit was able to induce apoptosis cell death
and suggests that more detailed studies should be carried out.
Keywords: Schinus terebinthifolius , peper tree, anti-neoplastic , cancer, apoptosis
Financial Support: FAPERJ, CNPq and CAPES.
Resumo:26-181
ANTIMYCOBACTERIAL AND ANTI-INFLAMMATORY ACTIVITY OF EXTRACTS AND HALOGENATED
SESQUITERPENES FROM THE BRAZILIAN RED ALGA LAURENCIA DENDROIDEA
Ventura, T. L. B. 1; Heggdorne-arajo, M. 1; Machado, F. L. 2; Lassounskaia, E. 1; Soares, A. R. 2;

Muzitano, M. F. 2
1
LBR/CBB - Universidade Estadual do Norte Fluminense, UENF
2
Universidade Federal do Rio de Janeiro/Maca, UFRJ/Maca
Objectives:
Tuberculosis (TB), caused mainly by Mycobacterium tuberculosis infection, remains a global health threat. Conventional
treatment involves the use of antimicrobial for a long term (6-9 months), leading to abandonment of treatment. In general, the
production of pro-inflammatory mediators is essential for inflammatory response to mycobacteria but the excessive production of
these and inappropriate activation of the immune system leading to severity of inflammation resulting from infection with
tuberculosis (J. Immunol.179: 3119, 2007). The marine organisms are a promising source of antimycobacterial compounds and
there is no study dealing with L.dendroidea antimycobaterial activity. The aim of this study was to find new immunotherapeutic
that beyond microbicide activity can act also in the exacerbated inflammation observed in TB.
Crude extracts from four populations of the red alga Laurencia dendroidea from Southeastern Brazilian coast and three isolated
halogenated sesquiterpenes: (-)-elatol, obtusol and cartilagineol were evaluated for their antimycobacterial and anti-inflammatory
activity. Fifty microliters of suspension of Mycobacterium bovis BCG (1 106 CFU/well) were plated and incubated with 50L
of each sample [100, 20 e 4g/mL]. After 7 days, 10l of MTT [5mg/mL] was added, and 3 hours after, lyses buffer. The
modulation of inflammatory response was assessed through of the inhibiting production of nitric oxide (NO) and tumor necrosis
factor (TNF-) and their cytotoxicity. RAW 264.7 murine macrophages were stimulated by LPS [1g/mL] and incubated with
samples [100, 20 e 4g/mL], n = 3. The amount of NO produced and toxicity were determined by Griess method, and both LDH
and MTT after 24h. The ability to inhibit TNF- was assessed by cell viability (MTT method) of murine L929 fibroblasts
incubated for 24 h with 50l of supernatant from the culture of RAW 264.7. The mechanism of inhibition of NO production was
evaluated through western blot analysis. L. dendroidea extract (Angra dos Reis, RJ, Brasil) were the most active against
Mycobacterium (IC50 8.66 1.36 g/mL) and on the inhibition of inflammatory mediators production: NO (IC50 5.30 1.33
g/mL) and TNF-. In addition, for isolated compounds, obtusol (IC50 31.44 0.76 g/mL) was more active on Mycobacterial
growth and (-)-elatol (IC50 16.51 1.08) on inhibition of inflammatory mediators, especially NO. This last activity showed to be
not only due to NO scavenging but due to a specific inhibition of iNOS activity or expression. It was the first time that
antimycobacterial and anti-inflammatory activity were described for L. dendroidea. and for (-)-elatol, obtusol and cartilagineol.
Conclusions:
In conclusion, our results showed that extracts from Laurencia dendroidea. and isolated halogenated sesquiterpenes contributes
to an anti-tuberculosis profile that is promising in the search of new alternatives for tuberculosis treatment.
Keywords: Anti-inflammatory, Antimycobacterial, Laurencia dedroidea, Marine products
Financial Support: Capes, Faperj and UENF
Resumo:26-182
INVESTIGATION ANTIDIARRHOEAL ACTIVITY OF HYPTIS MACROSTACHYS BENTH. (LAMIACEAE) IN
MICE
Vasconcelos, M. A. 1; Clementino-neto, J. 1; Silva, A. D. S. 1; Silva, K. M. 1; Costa, V. C. O. 2; Tavares, J.

F. 2; Silva, M. S. 2; Cavalcante, F. A. 1
1
1Instituto de Cincias Biolgicas e da Sade/UFAL, ICBS
2
Laboratrio de Tecnologia Farmacutica/UFPB, LTF
Objectives:
as some species of Hyptis (Lamiaceae) are used in folk medicine for gastrointestinal disorders, we decided to investigate possible
toxic and antidiarrhoeal activities of ethanol extract obtained of leaves from Hyptis macrostachys (HM-EtOH) in mice.
Methods: Pharmacological behavioral screening and Acute toxicity: were used mice (n=6) treated with saline p.o. (10 mL/kg) or
HM-EtOH (2500 and 5000 mg/kg p.o. or 1000 and 2000 mg/kg i.p.), and several behavioral parameters were evaluated for 4h
and quantified the deaths number (72 h). Castor oil-induced diarrhoea: mice (n=6) were divided into negative control (10 mL/kg
saline), positive control (10 mg/kg loperamide) and test groups (HM-EtOH at various doses). Diarrhoea was induced by oral
administration of 0.01 mL castor oil/ animal gram. Total number of faecal output and number of wet faeces excreted by the
animals were recorded (4h). Normal intestinal transit: mice were divided into groups (n=6). Group 1 received saline p.o., group 2
was administered 2 mg/kg atropine p.o. (positive control) and the other groups were treated with HM-EtOH p.o. After 30 min,
standard charcoal meal (0.01mL/animal gram) were given to mice orally. Animals were sacrificed and the small intestine
immediately isolated and determined the distance traveled by marker. Intestinal fluid accumulation: mice were divided into
groups (n=6). Group 1 received saline; group 2 was administered 10 mg/kg loperamide and the other groups were administered
HM-EtOH at various doses. 2mL of castor oil/animal was administered, and half an hour later, the mice were euthanized, and the
fluid volume was measured. All the experimental protocols were submitted by Ethical Committee in Research of UFAL (Protocol
N 23065.006775/2011-09). Results: HM-EtOH did not induce behavioral changes, or induce death of animals showing absence
of acute toxicity. HM-EtOH produced a significant antidiarrhoeal activity (p < 0.05), both the frequency of defecation (ED50 =
248.0 41.0 mg/kg), and liquid faeces (ED50 = 201.8 21.7 mg/kg) in mice. However, until the dose of 500 mg/kg the HMEtOH did not significantly inhibited intestinal motility when compared to negative control. Differently, in the evaluation of
intestinal fluid accumulation HM-EtOH inhibited significantly (p < 0.05) the intestinal secretion (ED50 = 259,9 65,3 mg/kg).
Conclusions:
based on our results suggest that the HM-EtOH extract has antidiarrhoeal activity; and this effect seems to involve only changes
in intestinal secretion.
Keywords: Antidiarrhoeal, Hyptis, Lamiaceae, Toxic
Financial Support: PIBIC/CNPq/FAPEAL/UFAL
Resumo:26-183
ANTIMICROBIAL ACTIVITY OF AGONANDRA BRASILIENSIS AGAINST OPPORTUNISTIC HUMAN YEASTS.
Souza, J. M. ; Vieira, M. C. S. ; Mendes, V. A. ; Moura, S. V. D. ; Ramon, J. D. L. ; Vieira, J. C. S;

Campos, E. P. ; Teles, H. L. ; Goulart, L. S.
Instituto de Cincias Exatas e Naturais, UFMT
Objectives:
The aim of this investigation was to evaluate the antifungal activity of extracts and fractions from leaves and stem from
Agonandra brasiliensis (Pau-marfim) against opportunistic fungi species by using broth microdilution method

Powdered shade-dried plant material was divided into two parts, one part was macerated in water:methanol (1:9) solution while
the other in dichloromethane:ethanol (7:3) solution. The crude hydromethanolic extract was partitioned with hexane,
dichloromethane and water:methanol (4:6). The crude extracts and three fractions were tested at concentrations ranging from
50000 to 70 g/mL. It was included in this study C. albicans ATCC 10231, C. glabrata ATCC 90030, C. krusei ATCC 6258, C.
tropicalis ATCC 25922, C. parapsilosis ATCC 40058, C. dubliniensis ATCC 7978, C. neoformans ATCC 32264 and C. gattii
ATCC 56990 isolates. Antifungal activity tests were performed using broth microdilution method as described in the M27-A2
document of Clinical and Laboratory Standards Institute (CLSI). MIC values were calculated as the lowest concentration of the
test samples which shows a complete growth inhibition of the fungi strains. The crude dichloromethane:ethanol extract and
dichloromethane and hydroalcoholic fractions from leaves were inactive against all fungal species tested. The crude
dichloromethane:ethanol extract and dichloromethane fraction from stem were not active to yeasts studied. The crude
hidromethanolic extract and hexane fraction from leaves were weakly active with MIC values ranging from 5000 to 2500 g/mL
and 5000 to 150 g/mL respectively. Hidromethanolic extract and hexane fraction from stem inhibits the growth of opportunistic
fungi, except for C. glabrata, with MIC values of 5000 to 300 g/mL and 5000 to 150 g/mL respectively. Hydroalcoholic
fraction from stem was only active against C. Krusei (MIC = 300 g/mL ), C. tropicalis (MIC = 150 g/mL) and C. dubliniensis
(MIC = 150 g/mL). The strongest effects were observed for hexane fraction from leaves and hexane and hydroalcoholic
fractions from stem against C. tropicalis (MIC of all the fractions = 150 g/mL) and C. dubliniensis (MIC of all the fractions =
150 g/mL).
Conclusions:
The current study suggests that A. brasiliensis have antimicrobial activity against opportunistic yeasts. Evaluations of the crude
extracts and fractions against clinical isolates are also being conducted.
Keywords: Antifungal, Agonandra brasiliensis, Yeasts
Financial Support: Fundao de Amparo a Pesquisa do Estado de Mato Grosso (FAPEMAT).
Resumo:26-184
EVALUATION OF NITRIC OXIDE SCAVENGING ACTIVITY OF PSYCHOTRIA NUDA EXTRACT AND
FRACTIONS
Calixto, S. D. 1; Ventura, T. L. B. 1; Heggdorne-arajo, M. 1; Gomes, M. V. S. 2; Oliveira, P. F. 2; Muzitano,

M. F. 2,1
1
Universidade Estadual do Norte Fluminense Darcy Ribeiro, UENF
2
Universidade Federal do Rio de Janeiro - Maca, UFRJ-Maca
Objectives:
The free radical nitric oxide (NO) plays an important role in the immune system, controlling infectious disease, for example.
However, it could be associated with worsening of inflammatory and degenerative diseases. Inhibition of NO production as well
as the free radical scavenging activity may be associated with an improvement in pathological condition. Sodium nitroprusside
(SNP) is a chemical compound, sensitive to light, used as a source of nitric oxide, allowing the evaluation of antioxidant activity.
The objective of this study was to standardize antioxidant testing using SNP and verify if the plant source selected is able to
sequester the NO released in culture supernatants. The speciesPsychotria nuda was chosen because its reported anti-inflammatory
and anti-mycobacterial activity.

The crystals of SNP were diluted to a curve of concentration-dependence (5, 10, 15, 20 and 25 mM) in medium DMEM-F12 and
then incubated at 25C at different periods time (30, 60, 90, 120, 150 and 180 minutes). This process was also evaluated in the
presence or absence of light. The best concentration of SNP, and incubation time were used for the test with the extract and
fractions of P. nuda at concentrations of 4, 20, 100 and 500 g/mL. After incubation, nitrite concentration was determine
colorimetric by Griess method, in comparison to the standard curve and to the flavonoid rutin, known for its antioxidant activity.
It is observed that the presence of light during incubation is an important factor for the antioxidant assay with SNP, the
concentration of 10 mM in 90 minutes showed a good performance and was adopted in our methodology. The activity observed
for the ethanolic extract was significant also at the lower concentration (4 g/mL) scavenging 74.360.61 % of NO radical
generated. Fractions in hexane, dichloromethane, ethyl acetate and butanol showed similar activity (between 67-77 % of
antioxidant activity), but the residual aqueous fraction was less active (57.862.59%).
Conclusions:
It can be concluded that the presence of light during incubation is important for the release of NO, and the ability to sequester the
free radical NO is one of the properties that makes P. nuda a promising candidate to treat inflammatory processes.
Keywords: Free radical, Nitric oxide, Psychotria nuda, Sodium nitroprusside
Financial Support: UENF; CNPq and Faperj Jovens Talentos
Resumo:26-185
EVALUATION OF THE ANDROGENIC AND ANTI-ANDROGENIC ACTIVITY OF HYDROALCOHOLIC
EXTRACT FROM VITEX AGNUS-CASTUS IN RATS
Sousa, M. R. S. C. 1; Vasconcelos, E. A. F. 3; Oliveira, J. M. G. 2; Lopes, J. C. O. 4; Nascimento, L. F. M. 1;

S, M. C. 2; Sales, P. A. B. 1; Pereira, C. F. C. 1; Lopes, S. T. P. 1; Costa, A. P. R. 1,2
1
3
Departamento de Bioqumica e Farmacologia, UFPI
2
4
Setor de Patologia Clnica Veterinria, UFPI
Objectives:
The existence of androgenic activity in plants can represent in addition to potential new therapeutic indications, undesirable
effects on reproduction, especially inhibition of spermatogenesis in males and masculinization of females. Therefore, the
objective of this paper was to evaluate a possible anti-androgenic and androgenic activity of the extract of Vitex agnus-castus in
rats.
The hydroalcoholic extract from Vitex agnus-castus was prepared by maceration from the dried leaves and crushed with 70%
ethanol for five days. Then, it was concentrated on Rotavapor at 50 C and then placed in glass jars and stored in refrigerator.
Were used 64 adult male Wistar rats weighing 180-250 g, kept under 12 hours light / 12 hours darkness in a room with controlled
temperature and free access to food and water. To evaluate the possible androgenic or anti-androgenic activity, the rats underwent
orchiectomy. Before returning from anesthesia, the animals received a combination of penicillin and streptomycin with
diclofenac sodium, to prevent infection and postoperative pain. The animals were left to stand for 30 days for complete recovery
of the surgical procedure and then were divided into these groups (n = 8): G1 - received saline at dose of 1 mL/100 g body weight
(bw) via gastric (vg); G2 - received 1mL/100g pc of the extract at 250mg/mL vg; G3 - received 1mL/100g pc of the extract at
500mg/mL vg; G4 - received 1mL/100g pc of the extract at 750mg/mL vg; G5 - received testosterone solution (2500 g / mL) at
dose of 0.1 mL/100 pc subcutaneously (sc) and saline vg at dose of 1 mL/100g pc; G6 - received testosterone sc + 1mL/100g pc
of the extract at 250mg/mL vg; G7 - received testosterone sc + 1mL/100g pc of the extract at 500mg/mL vg; G8 - received
testosterone sc + 1mL/100g pc of the extract at 750mg/mL vg. After 32 days of treatment, the rats were euthanized by excess of
anesthesia and proceeded to collection and evaluation of the prostate, seminal vesicles and adrenal glands. Then statistical
analysis was performed, and the differences between means tested with one-way ANOVA and Tukey post-test (p
Conclusions:
The hydroalcoholic extract from Vitex agnus-castus did not show androgenic or anti-androgenic activity in any of the
concentrations administered.
Keywords: androgenic, testosterona, Vitex
Financial Support: PIBIC/ Universidade Federal do Piau
Resumo:26-186
ANTIMICROBIAL ACTIVITY OF ALPINIA SPECIOSA (K. SCHUM.) EXTRACTS ON DIFFERENT BACTERIAL
SPECIES
Fochat, R. C. ; Nicolau, R. D. S. ; Silva, A. F. ; Raposo, N. R. B.

NUPICS/Faculdade de Farmcia da UFJF, FF/UFJF
Objectives:
To verify the antimicrobial activity of different extracts obtained from Alpinia speciosa (K. Schum.) leaves on three bacterial
strains and to determinate a Minimum Inhibitory Concentration (MIC) of them.
The vegetal material was collected in April 2010 in the Forestry Garden of the Faculty of Pharmacy of the Federal University of
Juiz de Fora - MG. The hexanic, ethyl acetate and methanolic extracts were prepared by Soxhlet (8 hours) and the ethanolic and
aqueous extracts by dynamic maceration (72 hours). The antimicrobial activity of the extracts was tested on Kocuria rhizophila
ATCC 9341, Enterococcus faecalis ATCC 51299 and clinical origin Escherichia coli EPEC, using the microdilution in culture
medium technique. The extracts were tested from 0.313 to 5 mg.mL-1 concentrations and the MIC was determinated by
turbimetric assay. All analyses were performed in triplicate and chloramphenicol (0.025 to 250 g.mL-1) was used as reference
drug. It was observed a bacteriostatic activity of ethyl acetate (MIC=5.0 mg.mL-1) and ethanolic (MIC=2.5 mg.mL-1) extracts
and bactericidal activity (MIC=2.5 mg.mL-1) of aqueous extract on Kocuria rhizophila ATCC 9341. No extract showed
antimicrobial activity on Entereococcus faecalis ATCC 51299. Finally, it was found a bacteriostatic activity (MIC=1.25 mg.mL1) of hexanic extract on clinical origin Escherichia coli EPEC. Chloramphenicol showed bactericidal activity on all bacterial
strains tested: Kocuria rhizophila ATCC 9341 (MIC=2.5 g.mL-1) and MIC=250 g.mL-1 to the other ones.
Conclusions:
The results showed the antimicrobial potential of A. speciosa extracts on Kocuria rhizophila ATCC9341 and clinical origin
Escherichia coli EPEC. Further studies are needed to confirm the antimicrobial activity of this plant specie, with the
perspective of obtaining a new antibiotic of natural occurrence.

Keywords: Alpinia speciosa, ANTIMICROBIAL ACTIVITY, Escherichia coli, Enterococcus faecalis , Kocuria rhizophila
Resumo:26-187
WOUND HEALING ACTIVITY OF OILS FROM COPAIFERA OFFICINALIS L. AND ROSMARINUS OFFICINALIS
L. LABIATAE ASSOCIATED TO SOLANUM SESSILIFLORUM DUNAL
Venncio, R. P. ; Gonalves, K. M. ; Soldati, P. P. ; Gomes, T. B. B. ; Rocha, P. R. ; Raposo, N. R. B.

NUPICS/Faculdade de Farmcia, UFJF
Objectives:
To verify the wound healing activity of essential oils from Copaifera officinalis L. (copaiba) and Rosmarinus officinalis L.
Labiatae (rosemary) associated with aqueous extract (AE) from fruits of Solanum sessiliflorum Dunal (cubiu).
Two combinations were prepared and incorporated in Lanette cream: a) 1% copaiba oil + 5% AE cubiu; b) 1% rosemary oil + 5%
AE cubiu. The experimental model chosen consisted of the induction of dermal ulcers in New Zealander albino rabbits (males,
1.5 to 2.0 kg, n = 4), their back being divided into four quadrants. In quadrant 1 it was tested the cream containing 1% copaiba +
5% cubiu while in quadrant 2 the test was performed using 1% rosemary + 5% cubiu. In quadrant 3 the Lanette cream was
applied (negative control) while in quadrant 4 it was 1% silver sulfadiazine (positive control). The ulcers were analyzed based on
the contraction of the lesion area (cm2) during 10 days. After this period, a study was made on histological slides stained with
hematoxylin-eosin to compare the number of inflammatory cells, fibroblasts, blood vessels, collagen area and area of
extracellular matrix between the experimental groups. The analysis of variance (ANOVA) was performed followed by Tukey's
post hoc test. Through macroscopic evaluation it was verified that treatment with 1% copaiba + 5% cubiu showed a decrease of
37.1% on the extension of the lesions, whereas treatment with 1% rosemary + 5% cubiu increased it on 4.3%, both in relation to
the positive control. Only the combination of 1% copaiba + 5% cubiu presented the wound area smaller than the negative control
(22.9% of shrinkage). On microscopic examination it was observed that the number of inflammatory cells was not statistically
different between the treatments of interest, both being statistically similar to the negative control. Both treatments showed a
higher number of fibroblasts in relation to the positive control (p = 0.020 for both). However, these values were not statistically
superior to the Lanette cream. Regarding the blood vessels, it was observed that the treatment with 1% copaiba + 5% of cubiu
showed the best result and was statistically higher than controls (p < 0.001) and treatment with 1% rosemary + 5% cubiu (p =
0.001). This same treatment presented an area of collagen statistically similar to controls. However, it was observed a significant
difference between treatment with 1% copaiba + 5% of cubiu and positive control (p = 0.004). Finally, it can be verified that
treatments with 1% copaiba + 5% cubiu and 1% rosemary + 5% cubiu showed an area of extracellular matrix with greater
extension than the positive control (p < 0.05) and similar to the negative control.
Conclusions:
The analysis of the data suggests that the treatment with the formulation containing 1% copaiba oil + 5% AE cubiu has the
greatest wound healing potential. It is suggested to conduct further analysis.
Keywords: WOUND HEALING ACTIVITY, Copaifera officinalis L., Rosmarinus officinalisL.Labiatae
Financial Support: CAPES. FINEP. MUNDO ANIMAL
Resumo:26-188
DEPIGMENTING AND WOUND HEALING ACTIVITIES OF AQUEOUS EXTRACT OF SOLANUM
SESSILIFLORUM DUNAL
Fonseca, Z. R. Q. ; Soldati, P. P. ; Gonalves, K. M. ; Venncio, R. P. ; Silva, A. F. ; Raposo, N. R. B.

NIQUA - FACULDADE DE FARMCIA, UFJF
Objectives:
To evaluate the anti-tyrosinase and wound healing activities of the commercial lyophilized (Experimental Station Santa Luzia)
aqueous extract (AE) obtained from fruits of cubiu (Solanum sessiliflorum Dunal).
In order to determine the anti-tyrosinase activity, the inhibitory activity of the tyrosinase was evaluated, according to Braz J
Pharm Sci. 45:4, 715-721, 2009. The experimental method used to evaluate the cicatrizing activity was the induction of dermal
ulcers in New Zealander albino rabbits (males, 1.5 to 2.0 kg, n = 6), their back being divided into four quadrants. In quadrant 1 it
was tested the cream containing 5% AE cubiu, while in quadrant 2 it was tested the cream containing 10% AE cubiu. In quadrant
3 it was applied the Lanette cream (negative control) and in quadrant 4, the silver 1% sulfadiazine cream (positive control). The
macroscopic analysis of the ulcers was done based on the contraction of the lesion area (cm2) for 10 days. After this period, the
histologic study of the slides (stained with hematoxylin-eosin) was performed in order to compare the number of inflammatory
cells, fibroblasts, blood vessels, collagen area and extracellular matrix area between the experimental groups. The descriptive
statistics were performed, along with the analysis of variance (ANOVA) followed by Tukey's post hoc test. Data were analyzed
through the software SPSS version 14.0, assuming a significance level of p
Conclusions:
The observed depigmenting activity was moderate. Although the treatments tested in the evaluation of cicatrizing activity have
shown a positive result concerning the vascularization of the wounds, the conjoint analysis of the results suggests that this activity
was considered modest for both formulations containing Solanum sessiliflorum.
Keywords: anti-tyrosinase , cicatrizing activity , cubiu, depigmenting activity , Solanum sessiliflorum Dunal
Financial Support: CAPES. FAPEMIG.
Resumo:26-189
ANTI-INFLAMMATORY POTENTIAL OF HEXANIC EXTRACT AND OF ITS FR2 FRACTION FROM
PTERODON POLYGALAEOFLORUS EVALUATED ON THE AIR POUCH MODEL.
Vigliano, M. V. ; Justo, M. G. ; Pinto, F. D. A. ; Silva, G. P. ; Leal, N. R. F. ; Marques, P. R. ; Sabino, K. C.
C. ; Coelho, M. G. P.
Departamento de Bioquimica, Ibrag - Uerj
Objectives:
The crude alcoholic extracts obtained from Pterodon pubescens Benth. fruits are widely used in Brazilian folk medicine as antiinflammatory, analgesic, anti-rheumatic tonics and depurative preparations (Spindola et al., 2011). The aim of this work was to
study the anti-inflammatory effects of hexanic fractions from Pterodon polygalaeflorus Benth. fruits using an acute inflammation
model.
The hexanic extract of Pterodon polygalaeflorus fruit (RB 350278, Jardim Botnico, RJ) (HEPpg) was obtained by maceration on
hexane for 15 days and further evaporation under vacuum. HEPpg was applied on a silica gel 60 column yielding six fractions,
which were analyzed by gas chromatography, TLC on plates of Silica Gel 60 and HPLC-DAD, being the Fr2Ppg fraction
analyzed in this work. The air pouch model was induced in male SW mice (2535 g b.w., n= 5/group). The pouch was done by a
sterile air injection of 5 mL in the mouse back. Three days after this, the pouch was maintained by 3 mL injection of sterile air. In
the six day, one hour before the administration of carrageenan (1 mL into the cavity), different doses (0.02 and 0.2 mg/kg) of
HEPpg and Fr2Ppg (100 L), prepared with the vehicle (ethanol 15% 1.25% Tween-20) were administrated. One group received
the vehicle (control) and another was treated with the control drug indomethacin (10 mg/kg b.w.). After four hours of carrageenan
injection, the animals were killed and their cavities were exposed, the exsudates were collected and measured. The air pouch
tissues were removed for histological procedures. All procedures were approved by CEA-IBRAG committee/protocol 05/2009.
The saline and vehicle groups showed cell amount around 2 and 40 x 106 cells/ml, respectively. Indomethacin inhibited 75%;
both HEPpg doses exhibited inhibition around 70%; and the lower Fr2Ppg dose reduced the cell number in 54,5% in relation to
the control group (vehicle). The highest Fr2Ppg dose not showed inhibition in relation to control group. The treatment with
HEPpg and its Fr2Ppg fraction reduced the exsudates volume in relation to the control group. These results were confirmed by
the histological evaluation of the air pouches.
Conclusions:
In conclusion, the HEPpg and Fr2Ppg showed efficiency in a reduction of the inflammatory index.
Keywords: Inflamao, Air pouch, Produto natural, Pterodon polygalaeflorus, Sucupira
Financial Support: FAPERJ, CNPq, Capes
Resumo:26-190
EFFECT OF LIPIDS TOTAL CONTENT OF THE MICROALGA ISOCHRYSIS GALBANA IN K562 HUMAN
ERYTHROLEUKEMIA CELL LINE
Vasconcelos, R. O. ; Souza, M. M. D. ; Buffon, J. G. ; Furlong, E. B. ; Britto, V. O. ; Abreu, P. C. O. V. D. ;

Moraes, M. M. D. ; Votto, A. P. S. ; Trindade, G. S.
Instituto de Cincias Biolgicas - ICB, FURG
Objectives:
The aim of this study was identify the fatty acids composition of the marine microalga Isochrysis galbana (I. galbana) and
investigate the possible antitumoral role of lipids total content in K562 human erythroleukemia cell line.
The microalga was grown in Guillard's F/2 medium, maintained in erlenmeyers, under constant aeration, 12:12 photoperiod, at
20C and in 28 salinity until stationary phase of growth. The culture of I. galbana (7500 mL) was centrifuged at 2500 rpm
for 15 min. So, the biomass of the microalga was incubated at 60C for 48h and it was stored at -20C until the extraction of
lipids to be realized. The lipids total content of the microalga was extracted in chloroform and methanol (J. Biol. Chem. 226:497,
1957). An esterification method (Anal. Chem. 38:514, 1966,) was performed with boron trifluoride reagent to establish its profile
of fatty acids. K562 cell line was maintained in RPMI-1640 medium with -mercaptoethanol, supplemented with 10% fetal
bovine serum, 1% antibiotic and antimycotic in cell culture flasks at 37C. K562 cell line was centrifuged, washed twice with
PBS, suspense in medium without -mercaptoethanol (5x105 cells/mL) and incubated with 1, 10, 100 and 1000 g/mL of the
lipidic extract. The control cells received only the ethanol vehicle (0,25%). The viability of K562 cells exposed to lipidic extract
was measured through method of exclusion by trypan blue 0h, 24h, 48h and 72h after incubation at 37C (n=6). The lipids total
content of the microalga was 61%. Regarding the profile of fatty acids found, it was observed oleic (17,1%), followed -linolenic
(13,6%), docosahexaenoic and nervonic (11,2%), linoleic (10,9%), -linolenic (8%), cis-10-pentadecenoic (8,5%), palmitic
(8,5%), miristic (7,8%), margaroleic (3,7%), myristoleic (1,9%), gadoleic (1,5%), lignoceric (1,3%), tricosanoic (1,2%),
eicosapentaenoic (1,0%), palmitoleic (0,6%), margaric (0,6%), eicosatrienoic (0,6%), lauric (0,5%), eicosadienoic (0,5%), beenic
(0,5%) and erucic (0,5%) acids. Relating to the lipidic extract effect in K562 cell line, it was showed an increase in the cell
proliferation in the concentrations of 1 (124,667 5,965) and 100 g/mL (131,2 12,80391) compared with control cells (73,333
4,667) in 48h. On the other hand, in the highest concentration tested, it was demonstrated a cytotoxic effect in 48h (86,63
1,81) and in 72h (80,15 1,39) with respect to control cells (94,29 1,67 and 91,41 0,55) respectively.
Conclusions:
In conclusion, the present study evidenced that the microalga I. galbana could be considered not only as an important source of
omega 3 fatty acids, but also omega 6 and omega 9 in relation to unsaturatedsaturated fatty acids. Moreover, this work suggests
that extracted lipids of the microalga I. galbana may influence cellular processes, capable of modulating cancer cell growth and
altering the cellular viability in K562 human erythroleukemia cell line.
Keywords: Erythroleukemia, K562, Microalga, Isochrysis galbana, Lipids total content
Resumo:26-191
INVESTIGATION OF ANTIFUNGAL ACTIVITY OF ESSENTIAL OIL OF ORIGANUM VULGARE ON CLINICAL
STRAINS OF CANDIDA TROPICALIS
Mendes, J. M. 1; Guerra, F. Q. S. 1; Mota, K. S. L. 1; Sousa, J. P. 1; Meneze, E. A. 2; Cunha, F. A. 2; Lima,

E. O. 1
1
Cincias Farmacuticas/Universidade Federal da Paraiba, UFPB
2
Anlises Clnicas/Universidade Federal do Cear, UFC
Objectives:
This study aimed to evaluate the antifungal activity of essential oil of Origanum vulgare against Candida tropicalis and to
characterize the relationship between the concentration of this essential oil with the rate and extent of its antifungal activity.
This work analyzed the activity of the essential oil of O. vulgare against C. tropicalis by minimum inhibitory concentration
(MIC), the minimum fungicidal concentration (MFC) and time-kill. The strains of C. tropicalis tested belong to the collection of
the Mycology Laboratory, UFPB (ATCC 13803) and the clinical strains are part of the Collection of Laboratory of Microbiology
of Yeasts, College of Pharmacy, Federal University of Cear (LMY/CP/UFC). The MIC was determined by the microdilution
method. In a 96-wells plate was added Sauborand dextrose broth and essential oil of O.vulgare at 1024 to 1g/mL concentrations
and incubated at 35-37C for 24-48 hours. The MIC was determined as the lowest oil concentration that inhibited visible growth
of microorganisms. To determine the MFC, 10 L aliquots of cavities where there was no growth of microorganisms were
transferred to sterile microdilution plates containing 100 L/cavity of Sauborand dextrose broth (SDB). After incubation at 3537C for 24 hours. The CFM was considered the lowest concentration seeded plate with sauborand dextrose agar (SDA) in which
growth was less than 3 colony forming units (CFU) (Bol. Micol. 4, 77, 1989; Phytochem, 69, 1895, 2008). The time-kill
(Antimicrob. Agents Chemother. 41 (6), 1392-1395): Colonies of the strains CT 10 and ATCC 13803 derived from culture in
SDA were suspended in 0.9% NaCl and turbidity adjusted according to the 0.5 McFarland scale. Then, fungal suspension was
added to SDB with or without the essential oil in various appropriate concentrations. The initial inoculum contained 1 x 105
CFU/mL. Concentrations of essential oil of O. vulgare tested were: MIC/2, MIC, MICx2. These cultures were incubated at 37 C.
At various time periods (0, 2, 4, 8 and 24 hours) a 1L aliquot of each dilution was removed and sown on plates with SDA. Plates
were incubated at 37 C for 24-48 hours and the number of colony forming units (CFU) was counted. The log10 CFU/mL was
plotted on a graph against time to check the rate and extent of antifungal activity in various concentrations of essential oil.
Fungicidal activity was considered when there was a decrease greater than or equal to 3 log10CFU/mL of initial inoculum, which
results in 99.9% reduction or greater in CFU/mL in 24 - 48 hours compared with the initial inoculation. (Diagn. Micr. Infec. Dis.
26, 125-131.; Diagn. Micr. Infec. Dis. 36, 101-105.). The results of tests shows that the MIC 90% essential oil of O. vulgare was
256 g/mL. The MBC was 256 g/mL. In time-kill study was found that the essential oil of O. vulgare exert fungicidal activity,
because had reduction of less than 99.9% or 3log10 the number of CFU/mL of initial inoculum.
Conclusions:
The antifungal activity demonstrated by the essential oil of O. vulgare makes it a potential candidate as an agent in controlling
fungi growth that cause infections, mainly Candida not-albicans strains.
Keywords: Origanum vulgare, Candida tropicalis, Essential oil , Antifungal activity
Financial Support: CNPq, DCF/CCS/UFPB.
Resumo:26-192
EFFECT OF QUERCETIN ON K562 HUMAN ERYTHOLEUKEMIA CELL LINE
Marques, M. B. ; Martins, C. R. ; Votto, A. P. S.

Instituto de Cincias Biolgicas, FURG
Objectives:
The aim of this study was to evaluate the sensitivity of K562 human erythroleukemia cell line to different concentrations of
quercetin, a compound found in the onion (Allium cepa).
K562 cells were maintained in RPMI-1640 with 10% fetal bovine serum, 1% antibiotic and antimycotic, in cell culture flasks at
37 C. The viability of K562 cells exposed to quercetina was performed trough three independent experiments containing
triplicates of samples. For each experiment, the cells were centrifuged, washed twice with PBS, resuspended in medium without
&beta-mercaptoethanol to a final concentration of 5x105 cells/mL and treated with different concentrations of quercetin (5, 10,
50, 100, and 500 &mug/mL). Control group received the same volume of sterile water. Cell viability (%) was measured by
Trypan blue exclusion immediately, 24, 48 and 72 hours after incubation with quercetin. It was observed an decrease in the
viable cells number (x 104 cells/mL) treated with the concentration of 5 &mug/mL (130.67 7.91) without change in cell
viability after 72 hours of exposure when compared to the control (232.33 28.33), indicating an effect of inhibition of
proliferation. The concentrations of 10, 50, 100 and 500 &mug/mL, showed cytotoxic effect from 48 hours of exposure with
respect to the control (CV = 89.00 0.93), as indicated by the decrease in the viable cells number (56.89 7.36; 50.89 5.37;
59.00 3.87; 73.80 9.25 respectively) and cell viability (76.25 2.14; 73.63 2.26; 72.68 3.20; 69.18 4.10 respectively).
Conclusions:
In conclusion, the present study indicated that quercetin was able of causing proliferation inhibition and cytotoxic effects in K562
human erythrouleukemia cell line.
Keywords: Erytholeukemia, K562, quercetin, inhibition proliferation, cytotoxicity
Financial Support: FURG
Resumo:26-193
IDENTICATION OF CHEMICAL CONSTITUENTS OF BASIL (OCIMUM AMERICANUM L) ESSENCIAL OIL AND
THEIR EFFECTS ON NEUTROPHIL MIGRATION IN ZYMOZAN ARTHRITIS-INDUCED MODEL
Yamada, A. N. ; Grespan, R. ; Fachini, F. C. ; Estevo-silva, C. F. . ; Pinho, R. J. ; Kummer, R. ; Bersaniamado, C. A. B. ; Cuman, R. K. N.

DEPARTMENT OF PHARMACOLOGY AND THERAPEUTIC, UEM
Objectives:
The aim of this study was to identify the chemical components and investigate if Ocimum americanun L essential oil (OEO)
reduced the neutrophil migration on zymosan induced-arthritis model
The Ocimum americanun L was collected in the city of Maringa, Paran, Brazil, March-April 2010, after have been identified and
stored under the number 11,160. The OEO was obtained from fresh leaves by hydrodistillation in a Clevenger-type apparatus for
three hours at 70C. Sample of OEO was analyzed by Gas chromatography-mass spectrometry (GC/MS). The chemical analysis
of OEO by GC/MS identified seven chemicals in the sample, 1.8-cineole (major component), linalol, camphor, terpineol,
eugenol, caryophyllene and germacrene D. The activity of OEO was evaluated in arthritis induced by zymosan (Sigma, St. Louis,
MO). The Balb/c female mice, weighing 22 2 g, were housed at 22 2C, light: dark cycle of 12 hours, with free access to food
and water. The protocol (number 066/2010) regarding this study was approved by the ethical in animal research (CAEA/state
University Of Maringa). The animals received intra articular injection of zymosan (200 g/10L of saline), thirty minutes after
oral treatment with vehicle (1% Tween 80 solution), 50, 150 or 300mg/Kg of OEO dissolved in vehicle. The control group was
injected with saline in contra-lateral knee joint. After six hours, the animals were euthanized and the knee joint exposed by
surgical incision. The articular cavity was washing twice with 5L of PBS containing EDTA. The total leukocyte count was done
in Neubauer chambers under optical microscopy (Nikon Eclipse E-200), values were reported as the number of cells per cavity.
Data were presented as mean SEM. The means from different treatments were compared by ANOVA with Tukey correction. P
values less than < 0.05 were considered significant. A significant inhibition of neutrophil recruitment was observed with groups
of mice pretreated with OEO in doses of 150mg/kg (7.2 x 104 1.5 cells/cavity), and 300mg/kg (9.1 x 104 2.1 cells/cavity) but
not 50mg/kg (13.4 x 104 2.1 cells/cavity) when compared with animals pretreated with vehicle and injected with zymosan (15.5
x 104 5.4 cells/cavity). Saline was used as negative control (0.19 x 104 0.08 cells/cavity).
Conclusions:
The present study report that OEO reduced neutrophil migration on experimental arthritis model, suggesting an important role in
therapeutic immunointervention to inflammatory diseases.
Keywords: NEUTROPHIL MIGRATION , BASIL (Ocimum americanum L) , ESSENCIAL OIL , ARTHRITIS
Financial Support: CNPq, CAPES and FUNDAO ARACRIA
Resumo:26-194
NEUROPROTECTIVE EFFECT OF OCIMUM AMERICANUM ON CELL DAMAGE INDUCED BY HYDROGEN
PEROXIDE IN RAT HIPPOCAMPAL SLICES.
Vanzella, C. ; Lovatel, G. A. ; Bertoldi, K. ; Almeida, E. F. ; Spindler, C. ; Barbosa, J. ; Von Poser, G. L. ;

Siqueira, I. R.
Objectives:
Many species of the family Lamiaceae have been reported as source of innovative neuroprotective agents, based on
acetylcholinesterase inhibition, antioxidant and antiinflammatory properties. Previous studies in our laboratory showed that
ethanol extract of Ocimum americanum (Lamiaceae), known as basil, has a potent antioxidant and in vitro anticholinesterase
activities, besides improved memory parameters after supplementation for 15 days in mice. The aim of this study was to evaluate
the neuroprotective effect of the ethanol extract of Ocimum americanum on the susceptibility to cell damage induced by
hydrogen peroxide (H2O2) in hippocampal slices from young and aged rats.
Young and aged male Wistar rats (4 and 16 months) were killed by decapitation, brains were removed and the hippocampi
dissected out. Hippocampal slices of 400-M thickness were prepared using a McIlwain Tissue Chopper. These slices were
individually pre-incubated for 30 min in HEPES-saline buffer. The medium was replaced by fresh buffer and the slices were
incubated with different concentrations of the ethanol extract of Ocimum americanum (0, 0.1, and 1 g/mL) for 1 h at 35 C.
After, the medium was replaced by fresh buffer in the absence or presence of H2O2 (2 mM). Mitochondrial activity, an index of
cell viability, was assessed by reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The plasma
membrane damage was measured by release of lactate dehydrogenase, using a kit (LDH, DOLES Reagents, Goinia, Brazil). The
incubation with H2O2 significantly impaired cellular viability in hippocampal slices from young and aged rats (reduction about
30 and 40%, respectively) (ANOVA followed by Tukeys, 65.85.8, p=0.009; 58.518.6, p=0.0001; respectively). The ethanol
extract at 1 g/mL reverted the damage induced by H2O2 in hippocampal slices from young rats (increase about 30%), but not
from aged rats (ANOVA followed by Tukeys, 96.429.2 p=0.053; 63.810.7, p=0.992; respectively). Besides, incubation with
H2O2 enhanced LDH released to the incubation media by hippocampal slices from young rats (ANOVA followed by Tukeys,
125.67.4, p=0.001), while ethanol extract of Ocimum americanum at 1 g/mL reverted this parameter (ANOVA followed by
Tukeys, 102.92.5, p=0.009). Interestingly, the incubation with H2O2 did not increase the LDH release in hippocampal slices
from aged rats, what may be related to other finding that age per se enhanced this parameter (t-Test, t=-2.996, p=0.013). The
ethanol extract of Ocimum americanum at 1 g/mL reduced the LDH release (ANOVA followed by Tukeys, 67.812.7,
p=0.006). Results were expressed as percentage of control and 100% was considered sample without H2O2 and ethanol extract.
Conclusions:
Our results indicate that hippocampal slices from aged rats respond to H2O2 brain injury differently than those from young rats.
The ethanol extract of Ocimum americanum reverted membrane cell damage in hippocampal slices in both ages and was able to
reduce impaired cellular viability in young rats. Our data suggests that Ocimum americanum contains useful neuroprotective
compounds.
Keywords: ethanol extract, hippocampal slices, hydrogen peroxide, Ocimum americanum, rats
Financial Support: CNPq, FIPE/HCPA
Resumo:26-195
CHEMICAL ANALYSIS AND IMMUNOPHARMACOLOGICAL ACTIVITY OF AQUEOUS AND METHANOLIC
EXTRAXTS OF THE FRUITS FROM SCHINUS TEREBENTHIFOLIUS
Borges I. F. . J. C. 1; Bernardes, N. R. 1; Muzitano, M. F. 2; Oliveira, D. B. D. 1

1
Universidade Estatual do Norte Fluminense Darcy Ribeiro, UENF
2
Universidade Federal do Rio de Janeiro, UFRJ-Maca
Objectives:
The objective was to evaluate the inhibition of chronic inflammatory processes through modulation of human lymphocytes
proliferation using the aqueous and methanolic extract prepared from the fruits of S. terebenthifolius and to compare the chemical
profile of these extracts by High Performance Liquid Cromathography (HPLC).
This work complements the previous results of our group who showed that the extracts and fraction of Schinus terebenthifolius
are able to inhibit nitric oxide production in cultured macrophages. It is the first time that the suppressive action on lymphocyte
proliferation is described for Schinus genus. Human lymphocytes, stimulated with 5 ug/mL of phytohemagglutinin - PHA
(mitogen) and treated with aqueous or methanolic extract of fruits were cultured for 5 days. Cyclosporin A was used as a drug
control. The proliferation was assessed by MTT. The chemical profile of aqueous and methanolic extracts were analyzed by
HPLC and it was observed only the presence of flavonoids in the methanolic extract emphasizing that this extraction was better to
obtain these compounds of interest. Aqueous and methanolic extracts were also tested against the inhibition of human
lymphocyte proliferation (100 and 20 ug/mL). The aqueous extract showed no inhibition of proliferation in human lymphocytes.
However, methanolic extract showed similar inhibition when compared to Cyclosporin in high concentration (83.763.8 %). This
result suggest that the differential chemical composition of the methanol extract confers its anti-inflammatory action, since
literature data show the high biological potential of flavonoids, compounds found only in this extract. To examine the toxic effect
of the extracts, these were evaluated by LDH assay. It may be noted that both aqueous and methanol toxicity was around 50% at
the high concentration (58.71.9%), and they decrease toxicity at low concentrations (13.44.0%).
Conclusions:
In conclusion, methanol extract of the fruits of the pepper tree is more effective in inhibit proliferation of human lymphocytes
stimulated by PHA and the identification of flavonoids only in this extract suggests the role of this class in the activity. The
immunomodulatory properties of pepper tree extract could corroborate to its popular use as an anti-inflammatory plant and
functional food.
Keywords: Anti-inflammatory, HPLC , Lymphocytes, Schinus Terebenthifolius
Financial Support: UENF,FAPERJ,CNPQ
Resumo:26-196
EFFECT OF SALINE AND METHANOLIC EXTRACTS FROM BARK AND LEAVES OF MYRACRODRUON
URUNDEUVA ON SURVIVAL AND BEHAVIOR OF NASUTITERMES CORNIGER
Gomes, F. S. 1; Napoleo, T. H. 1; Xavier, H. S. 2; Coelho, L. C. B. B. 1; Paiva, P. M. G. 1

1
1Departamento de Bioqumica, CCB, UFPE, Recife, PE., UFPE
2
2Departamento de Cincias Farmacuticas, CCS., UFPE
Objectives:
Secondary metabolites (alkaloids, flavonoids, terpenoids, and other phenolic compounds) participate in insect-plant interactions
by their toxic, repellent or attractive properties. Lectins are proteins that recognize carbohydrates and have showed insecticidal
activity. Myracrodruon urundeuva bark and leaves contain lectins which are soluble in saline solution and showed termiticidal
activity on Nasutitermes corniger. This work reports the phytochemical analysis of methanolic and saline extracts from M.
urundeuva bark and leaves and the investigation of their effects on survival and behavior of N. corniger workers and soldiers.
Saline extract (SE) were obtained by homogenization of bark or leaf powder in 0.15 M NaCl (16 h at 4 C) followed by filtration
and centrifugation (9,000 g, 4 C, 15 min); the extracts were dialyzed against distilled water. Methanolic extracts (ME) were
obtained by mixing (3 h, 28 C) the tissue powder with methanol and then dried in rotary evaporator. The ME from bark and leaf
were then resuspended in distilled water. The classes of secondary metabolites were determined by thin layer chromatography
using adequate development systems and revealers. N. corniger (16 workers and 4 soldiers) were transferred to plastic cups
containing a paper disk soaked with 200 L of extract (1.0, 2.0 and 3.0 mg/mL, dry weight/volume). Distilled water was used as
negative control. Bioassay was achieved in quintuplicate. To evaluate repellent or attractive effect on termite behavior, Petri
plates were filled up with 2% agar solution and wells are made in agar by the removal of a one central and eight peripherical
cylinders. Filter paper disks soaked with 15 L of extracts or distilled water (negative control) were placed in peripherical wells.
Termites (16 workers and 4 soldiers) were then transferred to central well. Assay was made in triplicate and the following
parameters were observed: absence or presence of termites in peripherical wells, construction standards of tunnels in agar, and the
closing by insects of constructed galleries. TLC detected the presence of hydrolyzable tannins and flavonoids in SE and ME from
bark and leaves. Leucoanthocyanidin was also detected in bark ME while -sitosterol was found in ME from leaves. Leaf extracts
showed higher variability of flavonoids types than bark extracts. SE from leaves showed termiticidal activity on workers at 3
mg/mL. Leaf ME was attractive for termites since termites built tunnels in direction to wells containing leaf ME and remained in
these wells during all the experiment. Bark extracts showed no effect on termite survival and have no repellent or attractive
property.
Conclusions:
Methanolic extract from M. urundeuva leaves was rich in secondary metabolites but not termiticidal on N. corniger. Termite
behavior assay indicates that leaf secondary metabolites attracted the termites. Termiticidal activity detected in saline leaf extract
is probably linked to lectin presence.
Keywords: phytochemical analysis, termiticidal activity, bark, leaf, extracts
Financial Support: FACEPE, CNPq and CAPES
Resumo:26-197
CYTOTOXIC EFFECTS OF EUCALYPTUS BENTHAMII ESSENTIAL OIL AND ITS MAIN TERPENE
CONSTITUENTS ON TUMORAL CELL LINES
Dll-boscardin, P. M. 1; Nakashima, T. 1; Farago, P. V. 2; Kanunfre, C. C. 2

1
Programa de Ps-graduao em Cincias Farmacuticas, UFPR
2
Setor de Cincias Biolgicas e da Sade, UEPG
Objectives:
Eucalyptus essential oils are widely used in food, pharmaceutical, cosmetic and perfumery industries. Several works show their
antibacterial effects on pathogenic bacteria in the respiratory tract, antiinflammatory and analgesic properties and repellent
activity. Nevertheless, there are more than 700 species of Eucalyptus and only some dozens are used as sources of essential oils.
Eucalyptus benthamii Maiden et Cambage is commonly used for reforestation in Southern Brazil due its remarkable cold
tolerance. The major components of this essential oil were identified by GCMS as pinene, terpinene, aromadendrene,
globulol and terpinen4ol. Therefore the aim of this study was to investigate in vitro cytotoxic potential of this volatile oil as
well as the related terpenes pinene, terpinen4ol and terpinene on J774A.1 (murine macrophage tumor) and Jurkat cells (T
leukemia cells).
The leaves of E. benthamii were air dried and then hidrodistilled for 6 h using a Clevenger type apparatus. The compounds
pinene, terpinen4ol and terpinene were obtained from Sigma. In vitro cytotoxicity test was performed using MTT
(methyltetrazolium test). The cell lines were submitted to increasing concentrations of essential oils (3, 10, 30, 100 and 300
gmL). One exposure period of 24 h was chosen for determining the in vitro cytotoxicity of each material. Cytotoxicity was
expressed as the concentration of sample that inhibited 50% of cell growth (IC50). The results are shown as mean SD from
three independent experiments (n5). Both the volatile oil extracted from E. benthamii leaves and the related terpenes showed
some degree of toxicity against evaluated cells. Jurkat cells were more sensitive to the studied essential oil (IC50 77.55 7.25
gmL) when compared to the J77A.1 cells (IC50 211.46 5.86 gmL). The same behavior was observed for pinene,
terpinen4ol and terpinene in which Jurkat cells were more sensitive to these compounds (IC50 217.84 1.21 gmL; 69.91
2.66 gmL and 184.32 6.25 gmL, respectively) than J774A.1 cells. The terpenes pinene and terpinene showed no
activity against J77A.1 cells while terpinen4ol revealed 231.98 8.22 gmL as IC50 value.
Conclusions:
The obtained results indicate that the essential oil of E. benthamii and its related components have moderate and low cytotoxicity
against Jurkat and J774A.1 cells, respectively.
Keywords: cytotoxicity, essential oil, Eucalyptus, reforestation
Resumo:26-198
INVESTIGATION OF TOXIC AND ANTIDIARRHOEAL ACTIVITIES OF AERIAL PARTS FROM SOLANUM
PANICULATUM L. (SOLANACEAE) IN MICE
Neto, J. C. 1; Mendona, M. A. O. 1; Silva, A. D. S. 1; Silva, K. M. 1; Silva, P. C. B. 2; Silva, T. M. S. 3;

Cavalcante, F. A. 1
1
Instituto de Cincias Biolgicas e da Sade, UFAL
2
Ps-Graduao em Produtos Naturais e Sintticos Bioativos, UFPB
3
Departamento de Qumica, UFRPE
Objectives:
Solanum paniculatum L. is popularly known as jurubeba-verdadeira. Based on chemotaxonomic criteria, coupled with the fact
that several species of Solanum have antidiarrhoeal activity, we decided to investigate a possible toxic effect and antidiarrhoeal
activity of ethanol extract obtained from aerial parts of S. paniculatum (SP-EtOHp) in mice.
Methods: Pharmacological behavioral screening and Acute toxicity: were used mice (n=6) treated with saline p.o. (10
mL/kg) or SP-EtOHp (2500 and 5000 mg/kg p.o. or 1000 and 2000 mg/kg i.p.), and several behavioral parameters were evaluated
(for 4 hours) and quantified the number of deaths (for 14 days). Castor oil-induced diarrhoea: mice (n=6) were divided into
negative control (10 mL/kg saline), positive control (10 mg/kg loperamide) and test groups (SP-EtOHp at various doses).
Diarrhoea was induced by oral administration of 0.01 mL castor oil/ animal gram 30 min after the above treatments. The total
number of faecal output and number of wet faeces excreted by the animals were recorded (4h). Normal intestinal transit: mice
were divided into groups (n=6). Group 1 received saline p.o., group 2 was administered atropine 2 mg/kg p.o. (positive control)
and the others groups were treated with SP-EtOHp p.o. After 30 min, standard (5%) charcoal meal (0.01mL/animal gram) were
given to mice orally. Animals were sacrificed 30 min after administration of charcoal meal, the small intestine immediately
isolated and determined the distance traveled by the marker. All the experimental protocols were approved by Ethical Committee
in Research of UFAL (Protocol N 010489/2009-15). Results: SP-EtOHp did not induce behavioral changes, or induce death of
animals showing absence of acute toxicity. SP-EtOHp produced a significant antidiarrhoeal activity (p
Conclusions:
The results of this study suggest that the SP-EtOHp possesses antidiarrhoeal activity, and this effect does not involve changes in
intestinal motility, however other studies must be carried out to elucidate the mechanisms involved in this activity.
Keywords: Solanum paniculatum L., aerial parts, diarrhoea
Financial Support: PIBIC/CNPq/FAPEAL/UFAL.
Resumo:26-199
FUCOSE MOIETIES IS ESSENTIAL FOR THE ABILITY OF FUCOSYLATED CHONDROITIN SULFATE TO
INHIBIT MUSCLE DAMAGE INDUCED BY BOTHROPS JARARACUSSU VENOM
Machado, M. M. ; Strauch, M. A. ; Tomaz, M. A. ; Martins, V. V. ; Fonseca, R. J. C. ; Mouro, P. A. S. ;

Melo, P. A.
PPGFQM / UFRJ, UFRJ
Objectives:
Snakebites by Bothrops jararacussu snake induces intense local tissue damage. The venom contains a complex mixture of
enzymes and small peptides. Phospholipases A2 are enzymes present in the venom which are responsible for a wide range of
activities, such as myotoxicity, oedema, anticoagulant, hemolytic, neurotoxic and cardiotoxic effects (Toxicon 45, p.1147, 2005).
Some polyanions have been shown to present antivenom properties against this venom (Toxicon 31, p.285, 1993). A new natural
polyanion polysaccharide, named Fucosylated Chondroitin Sulfate (fucCS), has been isolated from the body wall of the sea
cucumber Ludwigothurea grisea, and it is involved in many biological activities (JBC 282 (20), p.14984, 2007). We assessed the
ability of fucCS and its analogue without fucose moieties, named Defucosylated Chondroitin Sulfate (defucCS) to antagonize the
muscle damage induced by B. jararacussu crude venom.
In vitro CK assays were performed with isolated mouse extensor digitorium longus (EDL) muscle bathed with venom alone (25
g/mL) or incubated with fucCS or defucCS (10-50 g/mL). In vivo experiments were performed by i.m. venom injection alone
or preincubated with fucCS or defucCS (1-10 mg/kg) and the plasma CK activity was evaluated before and 2 hours after injection
(1 mg/kg). The phospholipase and hyaluronidase activities were measured using turbidimetric methods. The CK content was
evaluated in EDL muscle after a perimuscular injection of venom (1 mg/kg). Histological sections were performed in EDL
muscle after crude venom perimuscular injection. All experiments were approved by the Committee of Animal Use of the Rio de
Janeiro Federal University (DFBCICB 026). It was observed that fucCS inhibits 75% of phospholipase venom activity with IC50
= 10 g/mL (n=10) and 100% of hyaluronidase activity with IC50 = 7 g/mL (n=5), in concentration-dependent manner.
Incubation of fucCS with the venom eliminates the increase of plasma CK, in vivo (n=4). The EDL muscle was preserved when
exposed to venom with fucCS in vitro (30 and 50 g/mL) (n=4). The reduction of the CK content was prevented by fucCS (1-10
mg/kg) (n=4). DeFucCS was unable to protect the phospholipase activity and miotoxicity in vivo and in vitro. Light microscopy
shows that fucCS can inhibit the muscle damage induced by the venom (n=4).
Conclusions:
FucCS was capable to inhibit venom activities related to tissue damage, although defucCS does not have this ability. These
results indicate that fucCS presents activity against Bothrops jararacussu venom and we believe that this antivenom activity may
be due to the interaction of negative charges of fucose moieties of fucCS with positively charges toxins present in this snake
venom, like others polyanions. Our study suggests that Fucosylate Chondroitin Sulfate is a potential new inhibitor of myotoxicity
induced by B. jararacussu venom.
Keywords: Bothrops jararacussu, Fucosylated chondroitin sulfate, Muscle damage, Polyanions, Creatino kinase
Financial Support: CNPq, CAPES, FAPERJ and PRONEX.
Resumo:26-200
EVALUATION OF POTENTIAL ANTIHYPERGLYCAEMIC FROM TERMINALIA CATAPPA LINN. IN RATS
NORMOGLYCEMIA
Ferreira, A. K. B. 1; Tenrio, E. P. 1; Costa, D. L. 2; Santana, A. E. G. 2; Humberto, M. M. D. S. 2; Ribeiro,

. A. N. 1
ESENFAR/ Universidade Federal de Alagoas, ESENFAR/UFAL

2
IQB/ Universidade Federal de Alagoas, IQB/UFAL
Objectives:
Terminalia catappa L., tree of Malaysia (Fitoterapia 75:253, 2004), is found on the Brazilian coast, including the state of
Alagoas, has high use in the population as a medicinal plant. The pharmacological evaluation of ethanol extract of stem bark
showed significant antidiabetic activity (Congresso brasileiro de farmacologia e teraputica experimental 42:39, 2010). Thus, this
study aimed to investigate the potential anti-hyperglycemic the ethanol extract of stem bark of Terminalia catappa L. (TCEE) in
normoglycemic rats.
Male Wistar rats, normoglycemic, weighing 200 to 270 g were divided randomly into three groups (n = 5): group 1 (control
glucose 2 g/Kg, v.o.), group 2 (glucose 2 g/Kg, v.o. + TCEE 200 mg/kg, v.o.) and group 3 (control glucose 2 g/Kg, v.o. +
glibenclamide 10 mg/Kg, v.o.). After fasting for 18 hours, drugs were administered, and then were followed for blood glucose
levels of animals from the venous flow, at 30, 60, 90, 120, 150, 180 and 240 minutes. The study was approved by the Ethics
Committee of the Federal University of Alagoas (010151/2008-82). The analysis was performed results start using values
expressed mean standard error of mean (SEM). *** p < 0.001 versus control glucose; *** p < 0.01 versus glibenclamide. The
glucose values obtained on consecutive times, groups 1, 2 and 3, respectively, were: 30 minutes (136.8 7.3, 87.4 1.8***,
100.8 3.1 mg/dL), 60 minutes (129.2 4.9, 82.0 4.6***, 89.8 4.1* mg/dL), 90 minutes (112.8 5.4, 76.4 4.8*** , 86.4
4.6* mg/dL) 120 minutes (106.4 5.6, 73.0 3.9**, 87.4 5.8 mg/dL) 150 minutes (98.6 6.8 and 70.4 2.0, 88.6 3.1
mg/dL) 180 minutes (90.2 5.9 77.8 4.0, 78.8 4, 6 mg/dL) and 240 minutes (108.8 8.9, 64.8 2.5***, 84.6 5.0 mg/dL).
Thus, the curve of blood glucose in group 1 shows a clear hyperglycemic effect that decreases with time due to the physiological
metabolism of glucose in the time of 240 minutes is a further increase in blood glucose, possibly due to metabolism of muscle
glycogen. In group 2 the glycemia curve shows no changes of hyperglycemia caused by large glucose administration, suggesting
that this form presents TCEE antihyperglycaemic effect due to stimulation of insulin release by the pancreas. In group 3, the
curve of glucose has a lower elevation when compared with group 1, but values are still s a lot higher than those in group 3.
Conclusions:
The results indicate that TCEE shows anti-hyperglycemic effect, which was higher than that of glibenclamide. However, further
studies are needed to define the mechanism of action of the extract.
Keywords: medicinal plants, rats, Terminalia catappa Linn., test oral glucose tolerance
Financial Support: CNPq, FAPEAL, PPSUS - MS
Resumo:26-201
PROTEOLYTIC ACTIVITY OF THE ENDOPHYTIC FUNGUS COLLETOTRICHUM GLOESPORIOIDES ISOLATED
OF THE LEAVES OF JUSTICIA PECTORALIS JACQ.
Silva, A. P. S. 2; Santos, I. P. 2; Azevedo, A. L. R. 1; Chagas, M. B. O. 2; Cavalcanti, M. S. 1; Lima, V. L. M.

2
1
Depart. of Mycology / Federal University of Pernambuco, UFPE

Depart. of Biochemistry / Federal University of Pernambuco, UFPE
Objectives:
The endophytic microorganisms are present within plant tissues and organs such as leaves, stems and roots of various plants, and
in some cases cause damage to the plant when environmental conditions and physiological status of the host become favorable.
These are of great importance in biotechnology by producing hydrolytic enzymes such as proteases, which are enzymes
responsible for hydrolytic cleavage of peptide bonds. Justicia pectoralis Jacq., chamba is popularly known as a medicinal herb
used in North and Northeast of Brazil. This study was done to analyze the proteolytic activity of Colletotrichum gloesporioides
(Penz) Sacc. endophytic fungus isolated of the chamba collected of the Guided Garden Center of Biological Sciences, Federal
University of Pernambuco, Recife, PE.
Leaves were collected and sent to Dept. Mycology of UFPE, where they were fragmented into leaves discs, 5 mm in diameter,
and carried out disinfection. Then the fragments were placed in Petri dishes containing potato-dextrose-agar (PDA) supplemented
with antibiotic (chloramphenicol 50 mg / L-1) at room temperature (28 2 C), where they were observed daily. After the
development of the colonies, the species identified as C. gloesporioides underwent the test for the detection of proteolytic activity
by hydrolysis of casein in milk-agar medium. Was used HCl at 5% for visualization of halos. The microorganisms tested showed
Enzyme Index (SI) ranging from 2.83 to 3.11, thus being considered to produce a protease according to parameters established by
Harrigan & McCanoe. Mt. Lab. Microb. Alim. y Prod. Lcteos. Espaa.361, 1979.
Conclusions:
All tested fungi showed potential in the production of proteolytic enzymes. These enzymes can be used in laundry detergents,
dairy and meat. Thus, one can conclude that the fungus C. gloesporioides isolated from the leaves of J. pectoralis is a new source
of these enzymes for these applications in biotechnological processes.
Keywords: Atividade proteoltica, Colletotrichum gloesporioides, Fungos endofticos, Justicia pectoralis
Resumo:26-202
THE EFFECT OF ESSENTIAL OIL FROM PTERODON EMARGINATUS SEEDS ON PLANTAR INCISIONINDUCED MECHANICAL HYPERNOCICEPTION IN MICE
Silva-jnior, D. C. 1; Nucci, C. 2; Mazzardo-martins, L. 4; Stramosk, J. 3; Santos, A. R. S. 4; Martins, D. F.

2,3,4
1
Curso de Medicina, Universidade do Sul de Santa Catarina, UNISUL

3
4
Objectives:
The economic burden of treating chronic pain that develops from acute pain in a 30 year-old individual over a lifetime could be
as much as $1 million. The prevention and effective relief of acute pain may improve clinical outcomes, avoid clinical
complications, save health care resources and improve quality of life. The Pterodon pubescens seeds are commercially available
at the Brazilian medicinal flora market and the crude alcoholic extract of this plant is used in folk medicine in anti-inflammatory,
analgesic, and anti-rheumatic preparations. Phytochemical studies of Pterodon genus have revealed the presence of alkaloids,
isoflavones, and diterpenes. In the current study, we investigated the effects of the essential oil of Pterodon emarginatus (EOP)
after plantar incision (PI) on a model of postoperative pain in mice.
Committee of the Universidade do Sul de Santa Catarina (UNISUL). Groups of mice with a plantar incision (PI) were subjected
to treatment with EOP (0.1-100mg/kg, intragastric, i.g.). Behavioral testing measurements were obtained from separate groups of
sham (n = 8), PI (n = 8) and PI + EOP (n = 8). 12 hours after PI mice were treated with EOP (0.1-100mg/kg, i.g.) and mechanical
hypernociception was examined before and at different time-points after treatment (1, 2, 3 and 4 hours). In separate experiment,
on day 1 after PI we evaluated the mechanical hypernociception 1h. after treatment with EOP (100mg/kg, i.g.) at each day after
PI for 6 days. The control group was treated only with vehicle (saline -NaCl 0.9%- and Tween 5%). The results show a
significant difference between groups (incision vs. incision + EOP, 100mg/kg, i.g.) for withdrawal response frequency occurred
at 1h. (p < 0.001) and 2h. (p < 0.001) after treatment with EOP. Furthermore, chronic treatment with EOP (100mg/k, i.g.)
inhibited mechanical hypernociception. This action started 1 day (inhibition of: 4711%) and remained significant up to 5 days
(inhibition of 6813%).
Conclusions:
The present results demonstrated that EOP decrease mechanical hypernociception induced by plantar incision in mice.
Keywords: Anti-hypernociception, Mice, Pterodon emarginatus, Postoperative pain , Surgery
Financial Support: UNISUL, UFSC, Capes.
Resumo:26-203
ACUTE TOXICITY EVALUATION OF HYDROALCOHOLIC EXTRACT OF ZORNIA BRASILIENSIS (FABACEAE)
Moreira, M. M. B. ; Batista, T. M. ; Lima, C. U. G. B. D. ; Tavares, J. F. ; Silva, M. S. ; Diniz, M. F. F. M. ;

Castello-branco, M. V. S.
Lab. de Tecnologia Farmacutica/Univers. Federal da Paraba, LTF/UFPB
Objectives:
Zornia brasiliensis (Fabaceae), popularly known as urinria, urinana and carrapicho, is used by folk medicine as a diuretic
(Oecol. Bras., 11 (3):323, 2007). Being the specie used for medicinal purposes, tests for toxicity evaluation of Z. brasiliensis are
essential. Few phytochemical and biological studies of Z. brasiliensis are reported (Fitoterapia, 78:215, 2007) . The present
investigation was carried out to evaluate the safety of crude hydroalcoholic extract of Z. brasiliensis (ZBE) determining its
potential toxicity. For that, we used two assays: acute toxicity assay in mice and brine shrimp test (Artemia salina).
To acute study, ZBE was administered in mice (oral) at dose of 2000 mg/kg (n=6 males, 6 females). General behavior adverse
effects and mortality (to calculate LD50) were determined for up to 14 days. Furthermore, indexes of organs (mg organ/g
animal), liver, heart, kidneys, spleen and thymus were evaluated to detect possible signs of toxicity (protocol number from the
Ethics Comitee/LTF: CEPA N 0509/09). The differences were compared by ANOVA one way followed by Tukey, using
GraphPad Prism. To brine shrimp test, eggs of A. salina were incubated in artificial sea water and light during 24 h for obtain the
larvae. ZBE (0-1000 g/mL) was added in tubes containing 10 nauplii. The set was incubated at artificial light for 24 h and then
the survivors larvae were counted to determine the LC50 (Lethal Concentration 50%) (Phytom., 8:395, 2001). Three replications
were done for each concentration and the experiment was repeated three times. LC50 values and its 95 % confidence intervals
were obtained by nonlinear regression, using GraphPad Prism. No death or behavioral changes was observed in experimental
animals (LD50 2000 mg/kg). In addition, no significant differences in water and food consumption were observed in female
group. However, there was a significant decrease in food consumption of males animals (24.85 2.17) compared to male control
group (42.13 3.63). No significant differences in the indexes of heart, liver, kidneys, spleen and thymus were observed, both in
males and females, when compared with the control group. The LC50 value obtained in brine shrimp test was 22.41 (21.46
22.87) g/mL. Considering the criterion of activity being LC50 values of A. salina.
Conclusions:
According to the data presented it is possible to infer that ZBE showed low toxicity when orally administered in mice and was
strongly bioactive against A. salina, which may be related to the presence of bioactive constituents in the plant.
Keywords: Acute toxicity, Artemia salina, Zornia brasiliensis
Resumo:26-204
EVALUATION OF BIOLOGIC ACTIVITIES OF TWO NEW DITERPENES ISOLATED FROM HYMENAEA
STIGONOCARPA.
Brasil, T. R. 1; Andreo, P. S. S. 1; Miranda, P. C. M. L. 3; Frauches, T. S. 1; Borges, F. V. 1; Kanashiro, M.

M. 1
1
CBB/LBR/Universidade Estadual do Norte Fluminense, UENF
2
CCT/LCQUI/Universidade Estadual do Norte Fluminense, UENF
3
IQ/Universidade Estadual de Campinas, UNICAMP
Objectives:
Terpens constitutes a wide class of natural products with pronounced biological activities such as antibacterial, antiinflammatory, anti-bleeding and cytotoxic effects. Therefore, this work aims to evaluate the biologic activities of two new
isolated compounds from Hymenaea stigonocarpa (jatob do cerrado).
Two substances with kaurane-type structure [metil kaur-16-en-18-oic (C1), metil kaur-15-en-18-oic (C2)] were purified from the
hexanic extract of Hymenaea stigonocarpa epicarp in a silica-gel chromatography columm impregnated with 10% of AgNO3. To
evaluate the biological activity in vitro, U937, H460, Colo 205 and Raw 264.7 cell lines were cultured (1x106/mL) in 96 wells
plate, and treated with compounds at different concentrations, where the higher concentration was 200M. Antiproliferative and
citotoxic activity of the compounds was determined by MTT assay and Lactate dehydrogenase-release assay (LDH) respectively,
after 24 and 48 hours of treatment. MTT assay showed that compounds were able to inhibit only Raw 264.7 cell line growing in a
dose-dependent manner. The cytotoxicity, measured by LDH quantification, showed that the compounds were not able to induce
enzyme release. Apoptosis analysis by fluorescence microscopy, using ethidium bromide and orange acridine double staining,
showed that compound 1 induced 60% of apoptosis cell death at the higher concentration and compound 2 induced 65% of
apoptosis at 25 M, achieving 100% at 200M. In order to evaluate the possible anti-inflammatory activity of the compounds, we
assayed the inhibition of nitric oxide (NO) production by Raw 264.7 cell line stimulated with LPS. NO concentration measured
by griess reaction showed that all compounds were able to inhibit NO production in a time and dose-dependent manner. Antiinflammatory activity was also tested in vivo by mixing 100, 200 and 300M of compound 1 and 5 g of Bothrops atrox snake
venom and injecting in Swiss mice hind paw. The assessment of edema and hemorrhage was made after 30min, 1, 2, 6, 8 and 24
hours. This experiment demonstrated a slight suppression of edema and hemorrhage at the highest concentration tested.
Conclusions:
This work showed that compounds [metil kaur-16-en-18-oic (C1) and metil kaur-15-en-18-oic (C2)] were able to inhibit NO
production in LPS stimulated Raw-264.7 cell line, suggesting a possible anti-inflammatory activity.
Keywords: ANTI-INFLAMMATORY, DITERPENES, HYMENAEA STIGONOCARPA., NATURAL PRODUCTS
Financial Support: CNPq, UENF, FAPERJ, UNICAMP
Resumo:26-205
STUDIES OF ANTIHYPERGLYCEMIC EFFECT OF MUSA PARADISIACA IN RATS
Guesser, S. M. 1; Postal, B. G. 1; Kappel, V. D. 1; Pereira, D. F. 1; Madoglio, F. A. 2; Reginatto, F. H. 2;

Silva, F. R. M. B. 1
1
Laboratrio de Hormnio & Transduo de Sinais, UFSC
2
Laboratrio de Farmacognosia, UFSC
Objectives:
Musa paradisiaca, commonly known as banana, is traditionally used as antidiabetic remedy in folk medicine. The aim of this
work was to study the acute effect of M. paradisiaca on oral glucose tolerance curve in hyperglycemic rats.
Leaves of M. paradisiaca were extracted by decoction (plant:ethanol 40oGL,1:10,wt/vol) for 10 min. After, the extract was
filtered and a part of this hydroethanolic extract was evaporated under reduced pressure to dryness, yielding the crude extract.
The other part was successively partitioned with n-butanol, yielding butanolic and aqueous residual fractions. Fasted normal
Wistar rats received 100 mg/kg p.o. of crude extract or 50 mg/kg of butanolic or aqueous residual fractions and after 30 min were
loaded with glucose (4 g/kg p.o.). The glycemia (mg/dL) was measured by glucose oxidase method at zero, 15, 30, 60 and 180
min after the glucose loading. The sulfonylurea, tolbutamide (100 mg/kg p.o), was used as a positive control as well as to
compare the effectiveness of acute effect of extract and fractions on glycemia. Oral administration of 100 mg/kg of the crude
extract resulted in a significant reduction in blood glucose at 15 min (20%;P0.01) and 30 min (15%;P0.05). Also, 50 mg/kg of
aqueous residue reduced the glycemia at 15 min (20%;P0.01) and 30 min (14%;P0.05). The butanolic fraction showed a
significant antihyperglycemic effect at 15 min (21%;P0.01), 30 min (18%;P0.01) and 60 min (14%;P0.05), this results was
similar to those observed to tolbutamide, that also showed the antihyperglycemic effect at these times.
Conclusions:
The butanolic fraction from M. paradisiaca leaves showed a significant potential antihyperglycemic effect by reducing
significantly serum glucose levels in hyperglycemic rats. Studies are underway in order to characterize the active compound and
further elucidate the mechanism.

Keywords: Musa paradisiaca , diabetes, antyhyperglycemic
Financial Support: REUNI/CAPES; CNPq; PIBIC-CNPq/UFSC; FAPESC.
Resumo:26-206
CYTOPROTECTIVE EFFECTS OF FLAVONOIDS RUTIN AND HESPERIDIN IN INSULIN-SECRETING CELLS
BRIN-BD11.
Felipe, E. T. 1; Sousa-maestri, J. 1; Braatz, S. M. 1; Carpinelli, A. R. 2; Newsholme, P. 3; Kanunfre, C. C. 1;

Oliveira-emilio, H. R. 1
1
Departamento de Biologia Geral , UEPG
2
Departamento de Fisiologia e Biofsica, USP
3
School of Biomolecular & Biomedical Science , UCD
Objectives:
Oxidative stress is considered an important process in the development of diabetes mellitus. It is well established that insulinsecreting cells have low expression and activity of antioxidant enzymes, making them highly susceptible to reactive species
damages. In order to prevent or reduce cell damage caused by oxidative stress, many studies have been conducted in an attempt
to find bioactive compounds with antioxidant potential. In this category are the flavonoids that are characterized by reactive
species neutralization (direct action) and action on endogenous antioxidant defenses. Therefore, the objective of this work was to
evaluate (in vitro) the potential cytoprotective of flavonoids rutin and hesperidin in insulin-secreting cells (BRIN-BD11) exposed
to oxidative stress induced by hydrogen peroxide.
The assessment of cell viability and the cytoprotective effects of flavonoids rutin and hesperidin were evaluated using the MTT
reduction method. BRIN-BD11 cells were treated with different concentrations of hydrogen peroxide (100, 200, 400, 500 and
600 M) for 2 hours. After analysis of MTT reduction, the concentration of hydrogen peroxide that inhibits approximately 50%
cell viability (IC50=350 M of hydrogen peroxide n=16) was defined. The possible cytoprotective effect of flavonoids rutin (10
M) and hesperidin (10 M) was evaluated by pretreatment of cells with these compounds (24 hours) and subsequent exposure of
cells to oxidative stress induced by hydrogen peroxide (350 M2 hours). It was not found significant cytoprotective effect of
flavonoids compared with control group (n7).The cells were also simultaneously exposed for 2 hours with hydrogen peroxide
(350 M) and flavonoid (isolated or combined). Cells were treated with 10 M of flavonoids rutin andor hesperidin and with
combined flavonoids: rutin (5 M) plus hesperidin (5 M) and rutin (3.33 M), hesperidin (3.33 M) and quercetin (3.33 M).
Only the combined flavonoids had significant effects on cell viability: rutin plus hesperidin (106.0 6.158%) and rutin,
hesperidin and quercetin (93.84 9.264%) compared to the control group (presence of hydrogen peroxide and absence of
flavonoids 67.00 9.971%) (n8). Data are expressed as mean standard deviation and analyzed by ANOVA (p<0.05).
Conclusions:
These results demonstrated that the cytoprotective activity of flavonoids rutin and hesperidin on insulin-secreting cells depends
on the synergism between these compounds.
Keywords: diabetes mellitus, flavonoid, hydrogen peroxide, insulin-secreting cell
Resumo:26-207
EVALUATION OF CARDIOVASCULAR RESPONSE INDUCED BY THE ETHANOL EXTRACT OF THE STEM
BARK OF ZANTHOXYLUM RHOIFOLIUM LAM. IN RATS
Tenrio, E. P. 1; Barros, A. K. F. 1; Filho, E. S. F. 2; Oliveira, A. P. 2; Ribeiro, E. A. N. 1

1
ESCOLA SUPERIOR DE ENFERMAGEM E FARMCIA , UFAL
2
NPPM, UFPI
Objectives:
Zanthoxylum rhoifolium Lam is a plant popularly known as "mamica-de-cadela" or "mamica-de-porca ", and it is used for the
treatment of gastrointestinal disorders. Some pharmacological activities have been reported in the literature, such as antibacterial,
antifungal (Pharm. biology 44:657, 2006), antinociceptive (J. Etnofarmacol. 129:227, 2010) and antitumour
(J.Pharmacol.576:180, 2007). This study evaluated the cardiovascular effects induced by ethanol extract of the stem of
Zanthoxylum rhoifolium (EEZR) in rats.
Normotensive Wistar rats and spontaneously hypertensive rats (270 - 300 g) were anesthetized with sodium pentobarbital and
polyethylene catheters were inserted into the abdominal aorta and inferior vena cava for measurements of blood pressure and
drug administration, respectively. The experiments were performed 24 hours after surgery. The study was approved by the ethics
committee of the Federal University of Piau (029/09). The results are presented as mean standard error of the mean. In nonanesthetized normotensive (n = 6), administration of the EEZR (0.5; 1; 5; 10; 20 and 30 mg/Kg; i.v., randomly) induced a
biphasic response in mean arterial pressure (MAP). The extract caused a decrease in MAP in the first four doses (-173, -162, 111 and -214 mmHg) and increase in MAP in the last doses (245 and 337 mmHg). These responses were accompanied by
changes in heart rate (-124; 80.5; 4415, -635, 231 and 172 bpm). In SHR non-anaesthetized rats (n = 6), the EEZR (0.5;
1; 5; 10; 20 and 30 mg/Kg; i.v., randomly) also induced a biphasic response in MAP (-155; -256; -222, 185, 191 and 257
mmHg) and changes in heart rate (51; -6016; -289, 324, 213 and 4716 bpm.). In the first four doses, the hypotensive
response to EEZR was significantly increased and heart rate was significantly attenuated, after nitric oxide (NO) synthase
blockade (L-NAME, 20 mg/Kg, i.v.) in normotensive rats.
Conclusions:
The results showed that the extract produced a hypotensive response in both normotensive rats and in hypertensive rats. The LNAME was also capable of significantly changing EEZR-induced effects in rats normotensive, suggesting that NO appears be
participating of this effect.
Keywords: Natural products, Zanthoxylum rhoifolium, cardiovascular system, rats
Financial Support: CNPq, FAPEAL, PPSUS - MS
Resumo:26-208
INVESTIGATION OF ANTIDIARRHOEAL ACTIVITY OF CROTON ECHIOIDES BAILL. (EUPHORBIACEAE) IN
MICE.
Silva, A. D. S. 1,2; Silva, K. M. E. 1; Neto, J. C. 1; Mendona, M. A. O. D. 1; Silva, T. M. S. 3; Cavalcante, F.

A. 1
1
Instituto de Cincias Biolgicas e da Sade, UFAL
2
centro de cincias da sade/Ps-Graduao em Produtos Natur, UFPB
3
Departamento de Qumica, UFRPE
Objectives:
as some species of Croton are used in folk medicine for diarrhea, we decided to investigate the possible antidiarroheal activity of
ethanol extract of stem barks from Croton echioides (CE-EtOH) in mice.
Castor oil-induced diarrhoea: mice (n=6) were divided into negative control (saline plus cremophor), positive control
(loperamide 10mg/kg) and test groups (CE-EtOH at various doses). Diarrhoea was induced by oral administration of 0.3 mL
castor oil/mice 30 min after the treatment. The total number of faecal output and wet faeces excreted by the animals were
recorded for 4h. Normal intestinal transit: animals were divided into 3 groups (n=6). Group 1 received saline plus cremophor
(negative control); group 2 were administered atropine 2 mg/kg, p.o. (positive control); and group 3 were administered CE-EtOH
at various doses. After 30 min, standard charcoal meal (0.3 mL/mice) were given to mice orally. Animals were sacrificed 30 min
after administration of charcoal meal and the small intestine immediately isolated. Intestinal fluid accumulation: mice were
divided into groups (n=6). Group 1 received saline plus cremophor; group 2 was administered loperamide 10 mg/kg p.o. and
the other groups were administered CE-EtOH at various doses. After 30 minutes, was administered 2mL of castor oil/animal.
Half an hour later, the mice were euthanized, and the fluid volume was measured. All the experimental protocols were approved
by Ethical Committee in Research of UFAL (Protocol N 010489/2009-15). Results CE-EtOH produced a significant
antidiarrhoeal activity (p
Conclusions:
diarrhoea is the frequent passage of wet faeces and it involves both an increase in the motility of the gastrointestinal tract, along
with increased secretion and decreased mucosal absorption of fluid. Therefore, the treatment of the diarrhoeal aims for, among
other objectives, to increase resistance to flow (segmental contraction, decrease propulsion and peristalsis) and to increase
mucosal absorption or to decrease secretion. In this context, the results of this study suggest that CE-EtOH has significant
antidiarrheal activity and this effect seems to involve only changes in intestinal secretion.
Keywords: Antidiarrhoeal activity, Croton echioides, Euphorbiaceae, Intestinal fluid, Normal intestinal transit
Financial Support: PIBIC/CNPq/UFAL/FAPEAL
Resumo:26-209
EFFECTS OF THE ETHANOLIC EXTRACT OF HYPTIS SUAVEOLENS (L.) POIT, LAMIACEAE, IN GASTRIC
ULCER BY ETHANOL AND ACIDIFIED ETHANOL
Jesus, N. Z. T. D. 1,2; Montenegro, C. A. 1; Leite, T. J. A. 1; Sales, I. R. P. 1; Tavares, J. F. 1; Batista, L. M. 1
Universidade Federal daParaba, UFPB

2
Universidade de Cuiab, UNIC
Objectives:
Hyptis suaveolens is used in traditional medicine as anti-inflammatory and antiulcer. Thus, the aim of this study was evaluated
the activity antiulcer gastric of Hyptis suaveolens in gastric ulcer models by ethanol and acidified ethanol.
The plant was collected in Pirizal District-MT and one sample of botanical material was deposited in Herbarium of UFMT
(voucher number 36.375), identified by Harri Lorenzi of Plantarum Institute. The ethanolic extract of Hyptis suaveolens (HSetOH) was produced by maceration. The experimental protocols were approved by the Institutional Committee for Ethics in
Animal Research of LTF/UFPB and they were registered under 301/10. After 24h of fasting, the animals (n=5-7) were
treated using 10 ml/kg of the following reactants: 8% Tween-80 solution(A), carbenoxolone -100mg/kg (B1) ,
Lanzoprazole 30mg/kg (B2), HS-etOH -62.5 (C), 125(D), 250(E) and 500 mg/kg(F). The antiulcer assays were performed
using the following protocols: absolute ethanol in male Wistar rats (180-250g) and 0.03M HCl/60% ethanol solution which
induced to the gastric ulcer in male Swiss mice (25-35g). The animals were sacrificed, the stomachs removed and the
Ulcerative Lesion Index (ULI) was determined and expressed as mean S.D using ANOVA followed by Dunnetts Test (p
Conclusions:
The ethanolic extract of the Hyptis suaveolens present antiulcer effects in the both models analyzed.
Keywords: antiulcer, Hyptis suaveolens, medicinal plants
Financial Support: FAPEMAT/ CAPES/CNPq
Resumo:26-210
ACTION OF INDOLE-3-ACETIC ACID ON STHAPHYLOCOCCUS AUREUS
Vaz, A. C. N. ; Pugine, S. M. P. ; Melo, M. P. D.

Departamento de Cincias Bsicas, FZEA, USP
Objectives:
Studies show that indole-3-acetic acid (IAA), a plant growth hormone, as antibacterial in vitro against various pathological
microorganisms including Staphylococcus aureus. S. aureus is a pathogen capable of causing a range of life-threatening and mild
diseases such as septicemia, meningitis, toxic shock syndrome and food poisoning. The virulence factors produced by S. aureus
can be found as biofilm production and enterotoxins production. The production of biofilm can be responsible for the persistence
of bacterial infections by increase the adherence of the microorganism and protection against antimicrobial substances. The
protein SEB, a Staphylococcal Enterotoxins produced by S. aureus, has also commonly implicated in Staphylococcal food
poisoning. The aim of this study was to evaluate the action of IAA on Staphylococcus aureus with different characteristic
conferred by biofilm production and the other by protein SEB production (gene seb presence). For this purpose S. aureus
incubated in absence or presence of IAA were evaluated upon membrane integrity and antioxidant enzymes activities as
superoxide dismutase (SOD) and catalase (CAT).

The strains ATCC 6538 (biofilm positive) and ATCC 14458 (gene seb presence) used in this study were donated by Fundao
Osvaldo Cruz, Brazil. Firstly, the strains lyophilized were resuspended in Tryptic Soy Broth (TSB) and incubated overnight at
37oC. S. aureus (3.0 x 108 CFU/mL) were incubated in TSB in absence (control) and presence IAA (30 mM) for 24 h at 37oC.
Then, the microorganism was obtained by centrifugation (pellet) and was used to membrane integrity assay and enzyme activities
determination. To membrane integrity assay the S. aureus (3.0 x 109 CFU/mL) resuspended in phosphate buffer saline (PBS)
were incubated with Hoechst-33342 (10 g/mL) and iodide propidium-342 (43 mol/mL), and evaluable by flow cytometry
(n=4). For enzyme activities determination, the microorganism was resuspended with lysostaphin (20 IU/mL), incubated at 37 oC,
centrifuged for 15 minutes at 13,000 x g and the supernatant was used for CAT and SOD activities determination by
spectrophotometer (n=7). CAT activity was determined by H 2O2 consumption at 240 nm. SOD activity was determined according
to the rate of reduction of Nitro Blue Tetrazolium (NBT) by superoxide anion (O2-) at 550 nm. Significance of differences was
calculated using ANOVA and Tukey test (P0.05). Results show that IAA reduces (0.47 log cycles) the membrane integrity in
both strains of S. aureus, compared with the respective control. IAA presence was shown an increase of 3.3 times the CAT
activities in strain ATCC 14458 and no effect was detected in ATCC 6538, compared with the respective control. The SOD
activity was increased 2.6 times in ATCC 6538 cultured in presence of IAA, compared with the control; however no alteration
was detected in ATCC14458.
Conclusions:
Conclusion: The IAA (30 mM) altered the membrane integrity and antioxidant enzyme activity of the S. aureus with virulence
factors as biofilm production and presence of seb gene.
Keywords: biofilm, enterotoxin, auxin, antioxidant, membrane
Financial Support: FAPESP for grants (2008/10412-6) and CAPES for PhD scholarship.
Resumo:26-211
ANTIMICROBIAL AND ANTIOXIDANT ACTIVITY OF ETHANOL EXTRACT FROM BIDENS PILOSA LEAVES
Soldati, P. P. ; Silva, A. F. D. ; Gonalves, K. M. ; Raposo, N. R. B.

NUPICS/Faculdade Farmcia, FF/UFJF
Objectives:
Evaluate the antioxidant and antimicrobial activity of ethanol extract (EE) from Bidens pilosa leaves against different microbial
strains.
The plant material (50 grams from leaves) of Bidens pilosa was collected in September 2010 in the Forestry Garden of the
Faculty of Pharmacy of the Federal University of Juiz de Fora/MG. After drying, the EE was prepared by dynamic maceration for
72 h at room temperature. The extract was rotavaporated and dried at 40C. The antimicrobial activity of EE (0.313-5 mg/mL)
was evaluated by turbimetric assay, through which the minimum inhibitory concentration (MIC) was determined. The EE was
tested against six strains from the American Type Culture Collection (Candida albicans ATCC 10231, Enterococcus faecalis
ATCC 51299, Kocuria rhizophila ATCC 9341, Shigella sonnei ATCC 25931, Salmonella enterica serotype Thyphimurium
ATCC 14028 and Staphylococcus aureus ATCC 25923) and two clinical strains (Pseudomonas aeruginosa and enteropathogenic
Escherichia coli - EPEC). Chloramphenicol (0.025-250 g/mL) was used as reference drug for bacterial strains, while nistatin
(0.2-2000 U/mL) was used for the fungal strain. The antioxidant capacity was determined by 2,2-diphenyl-l-picrylhydrazyl
hydrate (DPPH) method (=510 nm). Ascorbic acid was used as antioxidant reference compound and both tests were performed
in triplicate. The EE showed bacteriostatic activity for Staphylococcus aureus ATCC 25923 (MIC=1.25 mg/mL) and for clinical
Pseudomonas aeruginosa (MIC= 5 mg/mL). Against Kocuria rhizophila ATCC 9341 bacteriostatic activity was found with MIC
= 0.625 mg/mL and bactericidal activity with MIC = 1.25 mg/mL. For the other strains, there was no antimicrobial activity. EE
showed antioxidant activity (IC50=16.1 g/mL) while the standard ascorbic acid was IC50=1.9g/mL.
Conclusions:
The results demonstrated the antioxidant and antimicrobial potential of ethanol extract from Bidens pilosa leaves. These findings
reinforce the need of more detailed studies about the chemical constituents of this plant and its activities, in order to enable
pharmacological applications for it.
Keywords: Antimicrobial activity, Antioxidant activity, Bidens pilosa, Pseudomonas aeruginosa , Staphylococcus aureus
Financial Support: CAPES. PROPESQ
Resumo:26-212
EFFECT OF HYDROALCOHOLIC EXTRACT OF NEEM (AZADIRACHTA INDICA) IN THE CONTROL OF
AEDES AEGYPTI.
Marinho, P. E. S. 1; Silva, K. P. D. 2; Barroso, W. A. 2; Camara, L. L. 2; Costa, M. C. P. 1; Silva, M. C. E. 1;

Borges, A. C. R. 2; Freire, S. M. D. F. 2; Camara, A. L. 2
1
Departamento de Quimica e Biologia, UEMA
2
Departamento de Ciencias Fisiologicas, UFMA
Objectives:
The aim of this study was to evaluate the effect of the hydroalcoholic extract of leaves of the Indian neem (Azadiractha indica A.
Juss) in the control of egg and larval stages of the A. aegypti.
The hydroalcoholic extract of leaves of the neem (EHAN) was prepared according with MATOS, 1997. The evaluation of
toxicity of the hydroalcoholic extract of neem on the eggs of A. aegypti was done in groups of 8 pallets at concentrations of 20%
and 25% of the extract, observed after 72 hours with water as control. The toxicity assessment EHAN on the larvae of A. aegypti,
was done in groups of 25 larvae, 3rd and 4th stage in small cups for each dose, and these were the same doses used for the eggs in
the total volume of 50 ml. To check the toxicity of the extract in adult rats was held the general test of pharmacological activity.
The general test of pharmacological activity was performed in adult rats of both sexes. The doses used in EHAN eggs and larvae
of A. aegypti were administered orally in groups of five rats for each dose, 20mg and 200m/kg rats, and a group of five rats as
control, which was treated with water. Tests with EHAN showed 100% inhibition of eggs hatching at the dose of 25% of the
extract and 98.43% of eggs hatched at a dose of 20% of the extract. It was observed that in control group the eggs hatched
normally (94.06%) after 72 hours. In tests using the same doses that eggs in larvae we observed a mortality of 76% to 25% of the
extract dose, which may make this compound a natural extract with larvae activity relevant, because have an average mortality
rate less than 50% after 24h of experiment (ref). The pharmacological tests showed that the administered doses have no toxic
effect on the rats, these results have is agreement with the data found in literature.
Conclusions:
These results showed that EHAN have ovicidal and larvicidal effect relevant at the doses used, non-toxic to mammals, thereby
demonstrating the feasibility of using this extract. From this statement one can get a viable product that can even be produced by
and used in population campaigns of the A. aegypti control, since this control is currently done only with chemicals that can be
harmful to other animals. Therefore already requested an Application for Invention: "Process for Preparation and Application of
Insecticides Compounds Obtained from the Hydroalcoholic Extract of Leaves of Neem (Azadirachta indica A. Juss) for the
Elimination of eggs and larvae of Aedes aegypti." Which is registered under number 000121. This project was approved by the
UEMA Ethics and Animal experimentation Comitee (CEEA), protocol number 014/2011. REFERENCES:MATOS, F.J.A.
Introduo Fitoqumica experimental. Fortaleza: UFC, 126p. 1997. MEDEIROS, Viviane Ferreira de. Potencial Larvicida de
Extratos de Plantas Regionais no Controle de Larvas de Aedes aegypti (Diptera: Culicidae). Natal: Universidade Federal do Rio
Grande do Norte, 2007.
Keywords: insecticide, neem, dengue, control, natural
Financial Support: BIC-UEMA Scholarship and Thanks to the Laboratory of Entomology - So Lus/MA.
Resumo:26-213
M.BRASILIENSE EXTRACT AND OIL INHIBIT PERIPHERICAL NOCICEPTION INDUCED BY CHEMISTRY
AGENTS IN MICE
Sawada, L. A. 1; Monteiro, V. S. C. 1; Sousa, T. S. 1; Conceio, L. R. V. 1; Silveira, A. J. A. 2; Nascimento,

J. L. M. 1; Bastos, G. N. T. 1
1
Universidade Federal Para, UFPA
2
Departamento de Quimica, UFPA
Objectives:
M. brasiliense (MB) is a medicinal plant found mainly in the North and Northeast areas of Brazil. In folk medicine, the seed is
used for faringites, diabetes, enterocolites, and as a mucolytic and antipyretic agent. So, this work aimed to investigate the
analgesic effect present in the MB seed extract and oil.
Swiss albino male mice (6 - 8 weeks), weighing 30 to 50 g, were divided randomly in groups (n=6). For the writhing test, mice
were divided in control (saline), 1, 5 and 10 mg/Kg concentrations of the MB extract and 10 mg/Kg of seed oil. One hour before
the test, the mice received the extract intraperitoneally. Writhing was induced by acetic acid (1%). The extract at 1, 5 and 10
mg/Kg reduce the number of writhing in 30%, 45% and 68%, respectively, showing a dose-dependent inhibition and the oil, at
10mg/Kg, administrated orally, presented a 52.1% inhibition. To evaluate central nociceptive effect, it was performed the hotplate test, utilizing the seed oil at 1, 5 and 10 mg/kg, which did not modified the latency time of termoception reaction compared
to the morphine group. Also, we investigated the MB oil composition through gas chromatography, that revealed high
concentrations of linoleic (32.82%) and oleic (21.79%) acid. All procedures involving animal care and experiment were
performed in accordance with guidelines of the ethical committee for research with experimental animals of UFPA(BIO001-09).
Conclusions:
The seed extract and oil promotes antinociception in visceral pain and the seed oil causes analgesic effect in a non-central
pathway. Also, there is a possibility that the analgesic activity is due to the oil components, since previous studies have shown
analgesic effects promoted by linoleic acid.

Keywords: analgesic, nociception, natural product
Financial Support: Cnpq, CAPES, UFPA.
Resumo:26-214
PSYCHOPHARMACOLOGICAL EVALUATION OF THE CRUDE ETHANOLIC EXTRACT OF HERISANTIA
CRISPA (L.) BRIZICKY.
Pereira, C. K. S. ; Oliveira, A. M. F. ; Penha, A. R. S. ; Timteo, R. N. P. D. ; Pinheiro, L. S. ; Teles, Y. C.

F. ; Matias, W. N. ; Souza, M. F. V. ; Almeida, R. N. ; Assis, T. S. D.
Laboratrio de Tecnologia Farmacutica/Univ. Federal Paraba, UFPB
Objectives:
Herissantia crispa (L.) Brizicky, popularly known as malvasco, belongs to the Malvaceae family (Rev. Bras. Farmacogn.,
18:472, 2008). Literature reports the use of species of this family in traditional medicine such as anti-fever, antiinflammatory e
diuretic (J Nat Prod., 53:1342, 1990). The aims of this work was to evaluate the psychopharmacological actions of Herissantia
crispa ethanolic crude extract (EEHc), in mice.
Swiss mice male weighing between 25 35 g (n= 6 ou 8), were divided into four groups: two test groups (500 or 800 mg/kg),
control (vehicle) and standard (specific for each test). The experimental protocol was approved by the Ethics Committee on
Animal Research of LTF / UFPB with registration N 0106/10. In the rotarod test, animals were placed on the rotating rod (7
r.p.m.), at the following time points: 30, 60 and 120 min; after treatment the remaining time on the bar was recorded. In the open
field test, animals were placed in the center of the field and allowed to explore freely for 5 minutes. The parameters observed
were: ambulation, defecation, rearing and grooming. In the test of elevated plus maze, the animals were placed on the apparatus
and observed the number of entries and time spent in open or closed arms for 5 min. During the test of seizures induced by
auricular electroshock the animals received an electroshock (0,5 mA, 150 pulses/s, 0,5 s) and tonic convulsions and death were
recorded. Regarding the sleeping time induced by thiopental test, the animals received a dose of 35 mg/kg (ip) of thiopental after
30 min of treatment and the latency and sleep time was recorded. EEHc treated groups did not show variation in the time spent on
the revolving bar in relation to control the rota-rod test. In the open field test there was a significant reduction of ambulation (500:
38,5 4,2; 800: 44,5 3,0; control: 82,7 5,3), defecation (500: 0,1 0,1; 800: 0,6 0,3; control: 3,0 0,8) and rearing (500:
10,1 3,5; 800: 15,0 1,5; control: 41,2 2,8) when compared to control, but there was no significant changes in grooming.
There was no significant change in the number of entries and time spent in the open and closed arms of elevated plus maze
compared to control. During the test of convulsion induced by electroshock there were no deaths and the animals treated with
EEHc showed tonic convulsions, similar to the control. The test of the potentiation of sleeping time, EEHc caused no significant
change in sleep latency but significantly increased the sleeping time induced by thiopental when compared to control (500: 53,4
11; 58,5 5,5; control: 11,1 2,9).
Conclusions:
EEHc does not show anxiolytic or anticonvulsant activity as shown by the elevated plus maze test or auricular seizures induced
by auricular electroschock, respectivelly. The muscle relaxing effect was discharted by the results obtained in the rota rod test.
However, EEHc presented sedative-hypnotic caractheristics, as evidenced in the open field and sleeping time induced by
thiopental test.
Keywords: Herissantia crispa, Psychopharmacology, sedative-hypnotic
Financial Support: UFPB, LTF, CAPES
Resumo:26-215
EFFECT OF THYMOL AND CARVACROL ON THE ELECTROPHYSIOLOGICAL PROPERTIES OF RAT
SENSORY NEURONS
Oliveira-abreu, K. ; Pimenta, P. V. C. L. ; Mendes, Y. C. ; Leal-cardoso, J. H. ; Barbosa, R.

Instituto Superior de Cincias Biomdicas, UECE
Objectives:
Thymol and its isomer carvacrol are components of essential oils from a wide variety of aromatic plants belonging to several
botanical families worldwide. Some plants that contain these components are found in Northeastern Brazil. Lippia sidoides
Cham, popularly known as "pepper-rosemary", contain thymol and carvacrol. These constituents are monoterpenes and phenolic
compounds and they have antibacterial, antifungal, and act on channels such as TRP (transient receptor channel). The aim of this
study was to investigate the effects of thymol and carvacrol on electrophysiological parameters of dorsal root ganglion (DRG)
somata of Wistar rats.
DRG were dissected from lumbar segments L4 and L5 of both sexes (180-200 gm). The use of animals was approved by local
ethics committee (CEUA - UECE) under the following protocol number: 06379067-0. The DRG were maintained between 18 C
and 24 C in modified Locke's solution (pH = 7.4). Changes in DRG membrane properties were monitored with sharp`
intracellular microelectrode technique (current clamp mode). We exposed DRG neurons to thymol (300M) and carvacrol
(300M) for 5 min and observed no statistically significant change on electrophysiological parameters analyzed. In experiments
with thymol control values for action potential (AP) amplitude, rate of AP rise (dV/dt) and rate of AP falling phase (dV/dt) were
respectively: 79 2.5 mV (34), 153 14.1 V/s (34) and -87 7.7 V/s (34). When the concentration of thymol was increased to 1
mM, AP parameters were significantly changed to respectively: 41 1.8 mV (n = 9), 68 26 V/s (9) and -44 16.6 V/s (9).
After exposure to 2mM thymol, AP values were changed to respectively: 27 0.3 mV (16), 31 8.5 V/s (16) and -33 7.8 V/s
(16). With respect to the actions of carvacrol, control values for AP parameters were, respectively: 78 1.8 mV (70), 115 5.7
V/s (70) and -106 11.9 V/s (70). After exposure to 600M carvacrol AP values changed to, respectively: 32 7.7 mV (28) and
48 13.0 V/s (28) and -48 11.8 V/s (28). At 3 mM, carvacrol completely blocked AP parameters. All these changes were
statistically different from control (p
Conclusions:
We conclude that carvacrol is more potent than thymol on AP parameters and the effects observed are probably due to a direct
action on Na+ channel.
Keywords: Thymol, Carvacrol, Current Clamp, Dorsal root ganglion
Financial Support: FUNCAP, CNPQ
Resumo:26-216
ANTITUMOR ACTIVITY OF PLANT SPECIES FROM THE SEMI ARID OF PARABA
Viana, W. P. ; Pessoa, D. R. ; Moreira, M. M. B. ; Pita, J. C. L. R. ; Silva, M. S. ; Tavares, J. F. ; Diniz, M.

F. F. M. ; Catello-branco, M. V. S.
Objectives:
For ethical reasons and financial resources, the use of in vitro tests is strongly recommended for the preliminary phase of testing,
using subsequently a smaller number of experimental animals (Toxic, 159: 135, 2001.). Fusaea longifolia, Xylopia
langsdorffiana, Pera leandrii, Nanuza plicata, Birsonima Gardneriana, Combretum duarteanum, Erythroxylum revoluntum,
Erytroxylum subrotundum, Lippia microphylla and Rollinia leptopetala are medicinal plants of native flora. The evaluation of
antitumor activity in vitro against malignant tumor cells of sarcoma 180 (S180) line has been widely used as a screening tool for
in vivo tests. Then, the objective of this work was evaluated the antitumor activity of these species on S180 cells using trypan
blue exclusion test.
To in vitro antitumor activity, S180 cells were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum (FBS),
penicillin (100 IU/mL) and streptomycin (100 g/mL) in 5% CO2 at 37 C. The cells (2x105 cells/well) were seeded in 96-well
plates and incubated with different concentrations of each extract (0-3000 g/mL), dissolved in 2% DMSO for 24 h. We used the
ethanolic extract of all the species studied. Trypan blue solution (0.4% in phosphate buffer saline, PBS) and cell suspension were
mixed in equal volumes (10 L) and the number of cells was estimated using a hemocytometer. Cells stained blue were scored as
dead. The IC50 value and its 95 % confidence intervals were obtained by nonlinear regression, using GraphPad Prism. Fusaea
longifolia, Xylopia langsdorffiana, Pera leandrii, Nanuza plicata, Birsonima Gardneriana showed IC50 of 166.1(165.8 - 166.5),
1047 (1041 1053), >5000, and 398.5 (381.2 - 416.6), 102 (98.54 105.6) g/mL. The IC50 values for Combretum duarteanum,
Erythroxylum revoluntum, Erytroxylum subrotundum, Lippia microphylla, Rollinia leptopetala were 77.4 (77.26 77.50), 239.5
(236,2 242,8),158.1(156.8 159.5), 512.3 (512.2 512.4), 278.3 (277.0 279.7) g/mL, respectively. The data presented allow
us to infer that the ethanol extract of Combretum duarteanum was the most active, while the extract of Pera leandrii showed the
lowest activity.
Conclusions:
Therefore, we conclude that most of the extracts showed moderate antitumor activity and may be selected for further tests in vivo.
Keywords: Antitumor activity, sarcoma 180 , semi arid, trypan blue
Resumo:26-217
ACUTE TOXICITY OF ESSENTIAL OIL OF XYLOPIA LANGSDORFFIANA (ANNONACEAE) FRUITS IN MICE
Macdo, B. V. F. L. ; Moura, A. P. G. ; Sousa, T. K. G. ; Duarte, M. C. ; Pita, J. C. L. R. ; Tavares, J. F. ;

Silva, M. S. D. ; Diniz, M. D. F. F. M. ; Castello-branco, M. V. S.
Objectives:
Aim: Some plants have potentially dangerous substances and, for this reason, they must be used with care, respecting their
toxicological risks. Xylopia langsdorffiana St-Hil. & Tul. is a tree, 5-7 m high and popularly known in Northeast Brazil as
pimenteira da terra (Z. Naturforsch, 62:742, 2007). The phytochemical study of essential oil of X. langsdorffiana fruits
(O.E.X.) characterized substances with important biological activities as monoterpenes (66.6 %), sesquiterpenes (22.2 %) and
diterpenes (11,2 %). The present investigation was carried out to evaluate the acute toxicity of O.E.X. determining its potential
toxicity after administration in mice.
Methods and Results: For the acute study, the O.E.X. was administered in mice (i.p.) at doses of 250, 375 and 500 mg/kg (n=6
males, 6 females). General behavior adverse effects and mortality (to calculate LD50) were determined for up to 14 days.
Furthermore, indexes of organs (mg organ/g animal), liver, heart, kidneys, spleen and thymus were evaluated to detect possible
signs of toxicity. The differences were compared by ANOVA one way followed by Tukey, using GraphPad Prism. The adverse
effects as well as lethality increased progressively with increasing doses (no deaths at 250 mg/kg to 11 deaths at 500 mg/kg).
Some adverse effects, such as ptosis, hyperactivity, aggressiveness were observed immediately after the i.p. administration, and
were more pronounced at higher doses, both in males and females. The highest dose showed sedation after 30 minutes of
treatment in females. The acute toxicity data indicated that the calculated LD50 value (J. Pharm. and Exp. Therap. 96:99, 1949)
for the i.p. doses of the O.E.X. was 351.09 mg/kg. No significant differences in the indexes of heart, liver, kidneys, spleen and
thymus were observed in males, when compared with the control group. However, the indexes of spleen and thymus increased
after the treatment with 250 mg/kg of O.E.X. in females. In addition, there was a decrease in body weight in all the animals
treated with 250 mg/kg of O.E.X.
Conclusions:
Conclusion: According to the data presented it is possible to infer that O.E.X. showed central toxic effects. The LD50 value of
351.09 mg/kg was used to direct the choice of safe doses to be tested in pharmacological assays to in vivo antitumor activity
evaluation. Repeated-dose toxicity studies are being conducted.
Keywords: ACUTE TOXICITY, ANNONACEAE, ESSENTIAL OIL, XYLOPIA LANGSDORFFIANA
Financial Support: Sources of research support: CNPq
Resumo:26-218
EFFECTS OF SEBASTIANA HISPIDA ON THE SPLEEN OF RATS INOCULATED WITH BOTHROPS MOOJENI
SNAKE VENOM
Duarte, P. O. 1; Almeida, M. F. 1; Matias, R. 2; Silva, K. 2; Dourado, D. M. 1

1
Laboratrio de Toxinologia e Plantas Medicinais, UAU
2
Laboratrio de Produtos Naturais, UAU
Objectives:
The objective of this work was to verify the effects of aqueous extracts from Sebastiana hispida on the spleen of Wistar rats,
inoculated with crude venom from Bothrops moojeni.
For the experiment, 36 male rats, Wistar strain, were used, with an average weight of 250g, obtained from Anhanguera Uniderp
University vivarium. The animals were separated into three groups: Control (SS, n=4), injected with sterile saline solution at
0.9%; Cv (n=4), injected with crude B. moojeni venom solution (40 g /ml); ShExt (n=4), treated with an aqueous extract of S.
hispida (40 g/ml) administered by intraperitoneal (IP) injection. The animals were injected with venom diluted at the moment of
use, by intramuscular (IM) injection into the gastrocnemius muscle. Then, the animals were anesthetized with Zoletil (IP) 0.1
ml/100g and the organ was removed, weighed, measured and immersed into buffered formaldehyde solution at 10% for 24 hours,
processed (alcohol and xylol), paraffin embedded, sectioned on a microtome (5 m) and stained with Haematoxylin and Eosin
(HE). This project was evaluated by the CEUA (Comisso de tica no Uso de Animais Animal Care and Use Ethics
Committee) under the protocol 64-006/09-MS. Alterations perceived in the spleen in the Cv group were the thickening and
inflammation of the capsule, hypertrophy, hyperplasia of both the white and red pulp, congestion, deposition of hemosiderin, and
depletion of periarteriolar lymphoid sheaths. For the ShExt group, the alterations were milder. At 3, 7 and 14 days, the spleen
weight for the SS group was: 0.380.71, 0.360.08, 0.300.03; for the Cv group: 0.860.06, 0.810.15, 0.750.06; and for the
ShExt group: 0.770.10, 0.630.04 and 0.630.10. Therefore, statistically, the Cv group showed heavier weights when compared
to the SS and ShExt groups.
Conclusions:
Histopathological alterations were evaluated in a semi-quantitative way, regarding the extension of alterations in all of the
histological slide, and it was attested that structural alterations were more evident at Cv group. As to the organs weight, it was
also verified that there was a volume increase in the Cv group. It may be concluded that, for the ShExt group, the venom effects
on the organ were diminished when treated with the plant extract.
Keywords: Histopathologic, herbal medicine, inflammation, water extract
Financial Support: MCT; INAU; CPP; FUNDECT and Anhanguera Uniderp University.
Resumo:26-219
ANTITUMOR ACTIVITY OF CRUDE HIDROALCOOLIC EXTRACT OF ROLLINIA LEPTOPETALA
(ANNONACEAE)
Mangueira, V. M. ; Batista, T. M. ; Lima, C. U. G. B. ; Pita, J. C. L. R. ; Moreira, M. M. B. ; Tavares, J. F. ;

Costa, V. C. O. ; Queiroga, C. S. ; Diniz, M. F. F. M. ; Castello-branco, M. V. S.
Objectives:
To evaluate the antitumor activity of crude hidroalcoolic extract of Rollinia leptopetala (RLE) we used in vitro and in vivo
assays. R. leptopetala, popularly known as "pinha-brava", is a plant endemic on Brazil and used by folk medicine as digestive and
against tumors and inflammations (Rev. Bras. Farmacog., 17:114, 2007).

To in vitro antitumor activity, sarcoma 180 cells (S180) were cultured in RPMI1640 medium supplemented with 10% fetal
bovine serum (FBS), penicillin (100 IU/mL) and streptomycin (100 g/mL) in 5% CO2 at 37 C. The cells (2x105 cells/well)
were seeded in 96-well plates and incubated with different concentrations of RLE (0-3000 g/mL), dissolved in DMSO, for 24 h.
The final content of DMSO in test medium and control was 0.1 %. The cultures were inspected for morphological alterations.
Tripan blue solution (0.4% in phosphate buffer saline, PBS) and cell suspension were mixed in equal volumes (10 L) and the
number of cells was estimated using a hemocytometer. Cells stained blue were scored as dead. The IC50 value and its 95 %
confidence intervals were obtained by nonlinear regression, using GraphPad Prism. Eight-day-old Sarcoma 180 ascites tumor
cells (25 x 106 cells/mL) were implanted subcutaneously into the left subaxilar region of Swiss mice (n=6) (J. Appl. Toxicol.,
28:599, 2008). One day after inoculation, the RLE (50, 100 and 150 mg/kg) or 5-fluorouracil (25 mg/kg) in Tween (10 %) was
administered (i.p.) for 7 days. There was a negative control. On day 8, all animals were euthanized and tumors excised and
weighed. Other parameters evaluated were: body weight, organ index and food and water consumption. The differences were
compared by ANOVA one way followed by Tukey. The RLE reduced S180 cells viability, with IC50 value 512.3 (512.2 512.4)
g/mL. The average tumor weight of the control mice was 2.196 0.235 g. The RLE reduced the tumor weight to 2.019 0.321
g, 1.345 0.243 g and 1.104 0.126 g at doses 50, 100 and 150 mg/kg, respectively. The average tumor weight of the 5-FU
treated mice was 0.44 0.08 g. These reductions gave inhibition ratios of 8.05, 38.72, 49.73 % and 80.06 % for mice treated with
50, 100 and 150 mg/kg of RL-EHB and 5-FU, 25 mg/kg, respectively. No significant changes in the body weight and food and
water consumption were observed. The index of the liver was reduced in animals treated with all the doses of RLE.
Conclusions:
Therefore, it is possible to infer that the RLE has significant in vivo antitumor activity and low toxicity on evaluated parameters,
which is an essential balance to its applicability as a pharmacological drug. Additional studies are being conducted to evaluate
liver damage.
Keywords: Antitumor activity , In vitro , in vivo , Rollinia leptopetala, Sarcoma 180
Resumo:26-220
ANTICONVULSANT EFFECT OF THE MONOTERPENE CARVEOL IN ANIMAL MODELS
Silva, E. V. B. 1; Nbrega, F. F. D. F. 1; Salvadori, M. G. S. S. 1; Sousa, D. P. D. 2; Almeida, R. N. 1

1
2
Departamento de Fisiologia/Universidade Federal de Sergipe, UFS
Objectives:
In recent years it has been reported several studies on the pharmacological properties of terpenes structurally simple. Most of
these substances are natural products abundant and well known, but its psychotherapy potential was underestimated. Carveol is a
natural unsaturated, monocyclic monoterpenoid alcohol that is a constituent of spearmint essential oil. In this study, we evaluated
the possible anticonvulsant activity of this monoterpene, using animal models classics.
Were used Swiss male mice (2535 g, n=8) maintained under controlled light and environment (12-h light/dark cycle, 21 1C)
with free access to food and water. All procedures were approved by the Ethics Committee of the Institution (protocol number:
0202/08). In the first test, seizures were induced by administration of Pentylenetetrazol (PTZ, 60 mg/kg, ip) and all the mice were
observed for at least 15min to detect the occurrence of the first episode of forelimb clonus (s). In the second test, seizures were
induced by electroconvulsive shock applied via ear-clip electrodes (0,5mA and 15 pulses/s for 0.5 s) and the percentage of
animals showing tonic convulsions was observed. In both experiments were administered doses of 125 and 250 mg/kg of the
drug-test, thirty minutes before induction of seizures. The results were analyzed by nonparametric ANOVA, followed by analysis
by the Dunnetts test or by Fishers exact test, according to the parameters. A probability of P
Conclusions:
Based on the results, we concluded that Carveol exhibits characteristics of drugs with anticonvulsant activity.
Keywords: Anticonvulsant, Carveol, Essential oil, Monoterpene, Seizures
Financial Support: UFPB, CNPq, RENORBIO
Resumo:26-221
OINTMENT ACTION OF SEBASTIANA HSPIDA PAX (EUPHORBIACEAE) IN THE WOUND HEALING IN RATS
Rizzi, E. S. ; Matias, R. ; Almeida, M. ; Aguiar, J. B. ; Torres,; Ferreira, M. B. ; Facco, G. G. ; Dourado, D.

M.
Universidade Anhanguera-Uniderp, UAU
Objectives:
To evaluate the healing potential of the ointment based on Sebastiana hspida in Wistar rats and to quantify the regression of
skin wounds in experimental groups.
The dried leaves of S. hspida, collected in Santa Emilia farm in Pantanal do Negro/MS, was used to prepare the methanol
extract, by maceration. 18 Wistar rats, adult, male (250g), divided into three groups (n=3): G1 (saline solution), G2 (2%
ointment + Carbopol Gel), G3 (2% ointment + lanolin/vaseline). The rats were anesthetized with sodium pentobarbital (10mg/kg)
administered by intraperitoneal (ip) and the trichotomy and antisepsis with 2% iodine alcohol was carried out at the surgical site.
For the skin excision the dorsal region was marked with a mold measuring 2 cm x 1 cm. After surgery the treatment was initiated
with 1g of the ointment in the injured area, once a day, in the period from 3 to 7 days of the experiment, after these periods, the
animals were subjected to euthanasia, using 50 mg / kg of pentobarbital sodium by intraperitoneal (ip). The histological pieces
were obtained through the removal of subcutaneous tissue from the injured area, then fixed in 4% paraformaldehyde, and stained
by Hematoxylin and Eosin (HE). This work was approved by the Ethics Committee (CEUA) number 770. For statistical analysis,
lesion measures were carried out daily from 3 to 7 days and registered (camera). The pictures were scanned and worked at
AutoCAD program, the data were statistically treated (ANOVA) and significant difference was not observed (P> 0.05), between
the experimental and control groups. Histological findings demonstrated that the first three days the skin of the experimental
groups presented ulceration areas, inflammatory infiltrate, predominantly neutrophils polymorphonuclear, necrosis, and
hyperemia. The dermis showed congested vessels and scarce inflammatory infiltrate. After seven days the G1 group presented
infiltrated inflammatory polymorphonuclear neutrophil. The G2 group observed beyond the mixed inflammatory infiltrate,
residual necrosis, hyperemia and fibroblast and in G3 group there was debris necrotic less thinner and inflammatory infiltrate less
intense, fibroblasts and vascular hyperemia.
Conclusions:
The group treated with the ointment-based Sebastiana hspida, provided better re-epithelialization, angiogenesis and activation
of granulation tissue than the other groups at day 7. It was observed that the vehicle used as lanolin / vaseline was more
expressive in the lesions than carbopol vehicle. Soon, the curative action in inflammation process and the lesion healing became
evident.
Keywords: angiogenesis , healing, re-epithelialization
Financial Support: MCT, CNPq; INAU, CPP, FUNDECT and the Anhanguera-Uniderp University
Resumo:26-222
STUDY OF LQB 118 AND LQB 149 PTEROCARPANS EFFECTS ON TUMOR
Martino, T. 1; Correia, R. R. 1; Jordo, F. C. 1; Coelho, M. G. P. 1; Costa, P. R. 2; Dias, A. G. 2; Sabino, K.

C. C. 1
1
Departamento de Bioqumica/UERJ , IBRAG
2
Departamento de Qumica Orgnica- UERJ, IQ
Objectives:
Pterocarpans are isoflavonoids isolated from Leguminosae, which show several bioprotective properties, including antioxidant,
antimutagenic, anticarcinogenic, antiproliferative, antiviral and antimicrobial effects (Phytochem. 55, 481-504, 2000; Life sci. 65,
337-353, 1999; Phamacol Rev. 52, 673-751, 2000). It has been demonstrated that pterocarpans induce cell cycle arrest and
apoptosis of leukemic cells. On the other hand, solid tumors like adenocarcinoma usually show high resistance to treatment with
cytotoxic drugs, reflecting a low response to chemotherapy treatment. Then, this work studied the in vitro antitumoral effects of
two new synthetic pterocarpans, the LQB 118 and LQB 149, on human lung (A549) and breast (MCF7) adenocarcinoma cells.
The different cells lines were expanded in RPMI medium containing 10% fetal bovine serum, plated (1 x 105cells/mL) in 96
wells microplates and incubated 24 h for adhesion, at 37oC and 5% CO2 atmosphere. Afterwards, the isoflavonoids LQB 118 and
LQB 149 were added or not to cultures, and the plates incubated for more 24 or 48 h. Cytotoxicity was determined by the MTT
assay, which determines the cell mitochondrial reduction activity, proportional to cell viability. The cell morphological features
were evaluated by light microscopy. Traditional chemotherapeutics (Doxorubicin, Etoposide and 5-fluorouracil) were used for
comparative studies. Cytotoxicity studies after 24 h culture showed that LQB 118 (2.5 g/mL) inhibited (p < 0.001) 55 + 22% of
breast cancer (MCF7) cells MRA, without inhibition on lung tumor (A549) cells. In 48 h cultures, the same substance (2.5
g/mL) increased inhibition (p < 0.001) of breast cancer cells MRA to 85 + 11%, while the lung cancer MRA showed no
inhibition. The cytotoxic effects of LQB 149 (2.5 g/mL) on both tumor cells were also tested at 24 h, showing no significant
inhibition of MCF7 or A549 cells MRA. In 48 h cultures, LQB 149 (2.5 g/mL) inhibited (p < 0.001) 76 + 11% of MCF7 cells
viability, while it did not inhibit A549 cells. The effects of other LQBs concentrations on these cells were also determined. The
chemotherapeutic drugs were tested in the more sensitive cell line (MCF7). Doxorubicin showed IC50 of 0.2 g/mL and 5Fluorouracil and etoposide showed IC50 of 22.7 g/mL and 108.1 g/mL, respectively, while LQB 118 and LQB 149, in the
same culture time (48h), showed IC50 of 1.9 g/mL and 2.3 g/mL, respectively. The LQB isoflavonoids cytotoxicity to breast
adenocarcinoma cells was accompanied by cell adhesion loss and cell size reduction.
Conclusions:
LQB 118 and LQB 149 isoflavonoids showed selective cytotoxic effects on adherent tumor cells of breast adenocarcinoma origin
(MCF7). As pterocarpans are estrogen-like, it is expected that their effects are greater on hormone sensitive cells, as here
demonstrated. Besides, both synthetics pterocarpans were more effective than some traditional chemotherapeutics (etoposide and
5-fluorouracil), suggesting them as potential alternatives of antineoplasic drugs. Experiments are in progress to determine the
mechanism of action of these samples
Keywords: A549/MCF7, Doxorubicin, Etoposide, Pterocarpans, 5-fluorouracil
Financial Support: CNPq, CAPES, FAPERJ, UERJ
Resumo:26-223
PURICATION OF A CHITIN-BINDING LECTIN FROM ALPINIA PURPURATA INFLORESCENCE
Marroqun, J. C. B. 1; Santos, N. D. L. 2; Brito, L. R. 1; Napoleo, T. H. 2; Pontual, E. V. 2; Coelho, L. C. B.

B. 2; Paiva, P. M. G. 2; Navarro, D. M. A. F. 1
1
Dep Qumica Fundamental, Universidade Federal de Pernambuco, UFPE
2
Dep. Bioqumica, Universidade Federal de Pernambuco, UFPE
Objectives:
Lectins are carbohydrate-binding proteins which differ from each other with respect to their molecular structures, carbohydrate
binding specificity and biological activities. Chitin-binding lectins participate in the defense mechanism of plants against
pathogens and have showed antibacterial, antifungal and insecticidal activities. Lectin activity (hemagglutinating activity) was
detected in saline extract from inflorescences of Alpinia purpurata (Zingiberaceae). The aim of this study was to purify the A.
purpurata inflorescence lectin (ApIL).
Proteins from A. purpurata inflorescence were extracted with 0.15 M NaCl and precipitated with ammonium sulfate (0-20%, 2040%, 40-60% and 60-80% saturation). The precipitated and supernatant fractions were dialyzed against distilled water (4 h) and
0.15 M NaCl (4 h) and evaluated for protein concentration and hemagglutinating activity (HA, titer) on rabbit erythrocytes treated
with glutaraldehyde. Specific HA was defined as the ratio between titer and protein concentration (mg/mL). HA inhibitory assay
was performed using 200 mM carbohydrates (fructose, galactose, glucose and N-acetylglucosamine). The 40% supernatant
fraction was loaded onto a chitin column equilibrated with 0.15 M NaCl. The adsorbed proteins eluted with 1.0 M acetic acid
(ApIL) were evaluated for protein concentration, HA and by polyacrylamide gel electrophoresis in presence of sodium dodecyl
sulfate (SDS-PAGE). ApIL HA was determined after heating (30 min) at 30-100 C. The supernatant 40% showed highest
specific HA (87,972) which was inhibited by glucose and N-acetylglucosamine. Chromatography profile of chitin column
showed a single active (specific HA: 29.681) protein (0.7 mg) peak (ApIL) eluted with 1.0 M acetic acid. SDS-PAGE revealed a
single polypeptide band for ApIL. Lectin activity was not altered after heating.
Conclusions:
A. purpurata inflorescences contain a chitin-binding and heat-stable lectin which can be isolated by one chromatographic step.
The characterization and evaluation of biological properties of ApIL are in progress.
Keywords: Alpinia purpurata, lectin, inflorescence, protein purification
Financial Support: FACEPE, CNPq and CAPES
Resumo:26-224
EFFECT OF HEXANE EXTRACT OF CALEA SERRATA ON ACETYLCHOLINESTERASE ACTIVITY OF THE
LARVAE OF RHIPICEPHALUS (BOOPHILUS) MICROPLUS.
Ribeiro, V. L. S. 1; Vanzella, C. 1; Santos, J. C. 2; Martins, J. R. S. 3; Poser, G. L. V. 2; Siqueira, I. R. 1,4

1
PPG em Cincias Biolgicas:BIOQUMICA, UFRGS
2
PPG em Cincias Farmacolgicas, UFRGS
3
Instituto de Pesquisas Veterinrias Desidrio Finamor, IPVDF
4
Departamento de Farmacologia, ICBS-UFRGS
Objectives:
Calea serrata Less. (Asteraceae) is an endemic species of south Brazil. Recently, n-hexane extract of this plant demonstrated
acaricide activity on the larvae of Rhipicephalus (Boophilus) microplus (Vet Parasitol.:151, 51, 2008), the cattle tick, a major
barrier to economic production of beef and dairy cattle. In arthropods, acetylcholinesterase (AChE) is a target for pesticides,
leadings to paralysis and death. Our work hypothesis was that components of Calea serrata could inhibit AChE in this larvae tick.
The objective of this work was to determine the effect of various concentrations of Calea serrata n-hexane extract on AChE
activity in larvae R. microplus.
Plant material (leaves and stems) of Calea serrata was collected in Porto Alegre, Rio Grande do Sul, Brazil. Air-dried and
powdered plant material was extracted by maceration with n-hexane, performing the solvent exchange until it become colorless.
After, the extract was evaporated to dryness under reduce pressure, treated with acetone and subsequently filtered and evaporated
affording an extract rich in chromenes and free of epicuticular waxes. Rhipicephalus microplus tick larvae were homogenized
(1:10) in 50 mM phosphate buffer pH 7.0 contained 0.5% of Triton-X 100 and centrifuged at 4 C. The supernatants were used as
the enzyme source. The enzyme activity was measured using the substrate acetylthiocholine and Ellman reagent (DTNB) (405
nm). The n-hexane extract of Calea serrata (final concentrations 1.5, 3 and 6 mg/mL) was incubated with the supernatants at 25C
for 60 min. AChE activity was estimated through differences in dA/min and calculated by comparison with control (without
Calea serrata). The n-hexane extract of Calea serrata inhibited (approximately 80%) the in vitro activity of AChE in larvae of R.
(B.) microplus at 6 mg/ml (0 mg/ml, median 100, 25%/75%-percentiles 100-100; 6 mg/ml, median 18.97, 25%/75%-percentiles
14.46-23.04; Kruskal-Wallis followed by Dunns, KW=16.922, p=0.0007).
Conclusions:
Our data indicate that the extract n-hexane of Calea serrata inhibits acetylcholinesterase in larvae of R. microplus, what can
contribute to, at least in part, acaricidal activity of this extract.
Keywords: ACETYLCHOLINESTERASE ACTIVITY , Calea serrata , HEXANE EXTRACT , LARVAE, Rhipicephalus
(Boophilus) microplus
Resumo:26-225
ANTIFUNGAL ACTIVITY OF THE ESSENTIAL OIL OF EUGENIA CARIOPHYLLATA ON STRAINS OF

CANDIDA TROPICALIS
Mendes, J. M. 1; Guerra, F. Q. S. 1; Sousa, J. P. 1; Mota, K. S. L. 1; Menezes, E. A. 2; Cunha, F. A. 2; Lima,

E. O. 1
1
Depto de Cincias Farmacuticas, UFPB
2
Depto de Anlises Clnicas e Toxicolgicas, UFC
Objectives:
This study aimed to evaluate the antifungal activity of essential oil of Origanum vulgare against Candida tropicalis and to
characterize the relationship between the concentration of this essential oil with the rate and extent of its antifungal activity.
This work analyzed the activity of the essential oil of O. vulgare against C. tropicalis by minimum inhibitory concentration
(MIC), the minimum fungicidal conecentration (MFC) and time-kill. The strains of C. tropicalis tested belong to the collection of
the Mycology Laboratory, UFPB (ATCC 13803) and the clinical strains are part of the Collection of Laboratory of Microbiology
of Yeasts, College of Pharmacy, Federal University of Cear (LMY/CP/UFC). The MIC was determined by the microdilution
method. In a 96-wells plate was added Sauborand dextrose broth and essential oil of O.vulgare at 1024 to 1g/mL concentrations
and incubated at 35-37C for 24-48 hours. The MIC was determined as the lowest oil concentration that inhibited visible growth
of microorganisms. To determine the MBC, 10 L aliquots of cavities where there was no growth of microorganisms were
transferred to sterile microdilution plates containing 100 L/cavity of Sauborand dextrose broth (SDB). After incubation at 3537C for 24 hours. The CFM was considered the lowest concentration seeded plate with ASD in which growth was less than 3
CFU (Bol. Micol. 4, 77, 1989; Phytochem, 69, 1895, 2008). The time-kill (Antimicrob. Agents Chemother. 41 (6), 1392-1395):
Colonies of the strains CT 10 and ATCC 13803 derived from culture in SDA were suspended in 0.9% NaCl and turbidity
adjusted according to the 0.5 McFarland scale. Then, fungal suspension was added to SDB with or without the essential oil in
various appropriate concentrations. The initial inoculum contained 1 x 105 CFU/mL. Concentrations of essential oil of O. vulgare
tested were: MIC/2, MIC, MICx2. These cultures were incubated at 37 C. At various time periods (0, 2, 4, 8 and 24 hours) a 1L
aliquot of each dilution was removed and sown on plates with Sauborand Dextrose agar. Plates were incubated at 37 C for 24-48
hours and the number of colony forming units (CFU) was counted. The log10 CFU/mL was plotted on a graph against time to
check the rate and extent of antifungal activity in various concentrations of essential oil. Fungicidal activity was considered when
there was a decrease greater than or equal to 3 log10CFU/mL of initial inoculum, which results in 99.9% reduction or greater in
CFU/mL in 24 - 48 hours compared with the initial inoculation. (Diagn. Micr. Infec. Dis. 26, 125-131.; Diagn. Micr. Infec. Dis.
36, 101-105.). The results of tests shows that the MIC 90% essential oil of O. vulgare was 256 g/mL. The MBC was 256
g/mL. In time-kill study was found that the essential oil of O. vulgare exert fungicidal activity, because had reduction of less
than 99.9% or 3log10 the number of CFU/mL of initial inoculum.
Conclusions:
The antifungal activity demonstrated by the essential oil of O. vulgaremakes it a potential candidate as an agent in controlling
fungi growth that cause infections, mainly Candida not-albicans strains.
Keywords: Candidiasis, Clinical strain, Clove, Fungicidal activity, Microdilution
Financial Support: CNPq, DCF/CCS/UFPB.
Resumo:26-226
ANTIFUNGAL ACTIVITY OF ALPHA-PINENE AND BETA-PINENE AGAINST CANDIDA ALBICANS STRAINS
Nbrega, F. M. 1; Lira, A. B. 1; Lima, I. O. 1; Lima, E. O. 1; Menezes, E. A. 2; Cunha, F. A. 2; Diniz, M. F.

F. M. 1; Souza, M. F. V. 1
1
DEPARTAMENTO DE CINCIAS FARMACUTICAS, CCS / UFPB
2
DEPARTAMENTO DE ANLISES CLNICAS E TOXICOLGICAS, UFC
Objectives:
The objective was to study the antifungal activity of -pinene and -pinene by minimum inhibitory concentration (MIC) and
determine which commercial antifungal will be used as a control in the research.
The strains of C. albicans tested belong the collection of the Mycology Laboratory, UFPB (ATCC 40042, ATCC 13803, ATCC
76485) and the clinical strains were isolated from blood (10), urine (1), respiratory tract (1), vaginal secretion (1) and are part of
the Collection of Laboratory of Microbiology of Yeasts, College of Pharmacy, Federal University of Cear (LMY/CF/UFC). The
MIC was determined by the microdilution method. Cultures of Candida albicans were placed on Sabouraud Dextrose Agar
(SDA) and incubated for 24-72 h at 37C. The inoculum was standardized according to the scale of 0.5 McFarland (1-5 x 106
CFU/mL). In a 96-well plate was added Sabouraud broth and -pinene, -pinene, amphotericin B, ketoconazole or flucytosine
concentrations of 1024 to 0,5 g/mL. In 24-48 h there was a visual observation of fungal growth. Negative control (without
drugs) was performed to confirm the cell viability (Planta Med, 64: 711, 1998; Mycol, 33: 75, 1999). The MIC value of -pinene
was 1024 g/mL (monoterpene -pinene was able to inhibit the growth of 19% strains) and -pinene was 1024 g/mL
(monoterpene -pinene, in turn, was able to inhibit the 44% strains).The amphotericin B showed the value MIC of 1 g/mL, the
ketoconazole, 128 g/mL and flucytosine showed values between 128 to 1024 g/mL (The commercial antifungals were able to
inhibit the growth of 100% strains).
Conclusions:
This study exposed that -pinene and -pinene had anti-Candida activity, but the MIC was higher when compared with
amphotericin B and ketoconazole. In addition to it, when compared the MIC values of the phytoconstituents and flucytosine the
results were similar.
Keywords: ALPHA-PINENE , ANTIFUNGAL ACTIVITY , BETA-PINENE , CANDIDA ALBICANS , MINIMUM
INHIBITORY CONCENTRATION
Financial Support: Conselho Nacional de Desenvolvimento Cientfico e Tecnolgico (CNPq).
Resumo:26-227
EFFECT OF CRUDE EXTRACT FROM MORINDA CITRIFOLIA (NONI) ON THE ONSET TIME AND DURATION
OF SLEEP INDUCED BY SODIUM THIOPENTAL IN MICE
Nascimento, L. F. M. 1; Fernandes, R. M. 1,3; Lira, S. R. S. 1; Fernandes, M. Z. L. C. M. 2,3; Oliveira, J. M.

G. 3; S, M. C. 3; Lopes, S. T. P. 1; Sousa, M. R. S. C. 1; Sales, P. A. B. 1; Oliveira, M. S. 1
1
2
Departamento de Bioqumica e Farmacologia, UFPI
3
Objectives:
Noni (Morinda citrifolia L.) is a fruit that came to Brazil as a raw material with strong commercial appeal due to all its beneficial
features. The popular use and same the traditional one are not sufficient to validate the herbal medicines as safe and effectives.
Thus the aim of this study was to evaluate the effects of Morinda citrifolia (noni) on the onset time and duration of sleep induced
by sodium thiopental in mice.
The crude extract was prepared with 8 kg of fruits of noni (M.citrifolia) mature and healthy, washed in running tap potable water,
followed by immersing them in stainless steel tanks with water chlorinated to 50 ppm for 10 minutes, then they were rinsed again
and dried. The fruits were cut in longitudinal and transverse slices, arranged in the shape of walls in a steel container with a
funnel attached to the bottom for the passage of the liquid, which after 48 hours, was stored in amber glass bottle and wrapped in
aluminum paper. Throughout the procedure, the material was covered, preventing oxidation and photosensitivity, avoiding loss of
properties. Were used albino mice (Mus musculus), adult male, weighing 20 g to 35 g, kept at the temperature of 22 to 25 C,
with light/dark cycles (12/12 h), receiving water and food ad libitum. To evaluate the onset time and duration of sleep were used
72 mice which were divided into 12 groups treated as follows: G1- treated with distilled water 0.1 mL/10g body weight (bw) +
sodium thiopental 50 mg/kg, both intraperitoneal (ip); G2 - distilled water orally (po) + thiopental ip; G3 - noni extract at
concentration of 100% (equivalent to 69mg/mL) at dose of 0.1mL/10g bw + thiopental, both ip; G4 - noni extract 100% vo +
thiopental ip; G5 - noni extract 50% + thiopental, both ip; G6 - noni extract 50% vo + thiopental ip; G7 - noni extract 25% +
thiopental, both ip; G8 - noni extract 25% vo + thiopental ip; G9 - noni extract 12.50% + thiopental, both ip; G10 - noni extract
12.50% vo + thiopental ip; G11 - noni extract 6.25% + thiopental, both ip; G12 - noni extract 6.25% vo + thiopental ip. The onset
time and duration of sleep data (minutes) were expressed as mean standard error of mean (SEM) group (n = 6) and analyzed by
Kruskal-Wallis test and Dunn post-test. Means accompanied by the standard error for onset time of sleep were: 2,33 0,21; 3,50
0,34; 3,17 0,17; 2,50 0,22; 2,50 0,22; 3,17 0,31; 2,83 0,31; 2,50 0,34; 3,67 1,28; 3,50 0,22; 3,33 0,21; 3,33
0,21 minutes for the groups G1 to G12, respectively. There was no statistically significant difference between the groups. The
results for sleep time induced by thiopental after administration of the vehicle and noni extract in the groups 1 to 12 were
expressed, respectively, as mean and SEM as follows: 11,50 2,13; 15,33 2,03; 32,50 7,99; 14,83 5,66; 89,00 13,16*;
46,67 32,70; 36,00 10,40; 64,50 36,67; 102,67 37,23*; 52,50 30,26; 19,33 5,71; 5,67 1,48* minutes. The group 12
showed a statistically significant difference compared to groups 5 and 9 (p <0.05).
Conclusions:
The groups treated with crude extract of Morinda citrifolia (noni), despite having increased sleep time in some concentrations,
did not differ statistically from control groups, suggesting no central nervous system depressant effect.
Keywords: noni, thiopental, latency
Financial Support: Universidade Federal do Piau
Resumo:26-228
EFFECTS OF MATE TEA (NATURAL AND DIET) IN HEALTH LUNGS OF AGED MICE
Cordeiro, N. C. 1; Lanzetti,m2; Barroso, M. V. 1; Pires, K. M. 1; Lopes, A. A. 2,1; Ribeiro, M. L. 3; Alves, J.

N. 2; Benjamin, C. F. 1; Porto, L. C. 2; Valenca, S. S. 1
1
2
Univerdade do Estado do Rio de Janeiro, UERJ
Universidade Sao francisco, USF
Objectives:
The Ilex paraguariensis herb has been commonly used in alternative medicine. After toasting process, the infusion known as mate
tea (MT) that contains many bioactive compounds that are cofactors in metabolic reactions. Our goal was to analyze the effects of
Mate Tea (natural [MTN] and diet [MTD]) in the lungs of mice during the aging process.
9-months-old male C57Bl/6 mice drank either MTN (n=11), MTD (n=10) or water (n=10) on a regular basis for 6 months.
Following euthanasia, lungs were removed and homogenized. The amount of reactive oxygen species (ROS) was measured. The
expression of key antioxidant enzymes (superoxide dismutase C SOD; catalase C CAT and glutathione peroxidase C GPx),
inflammatory cytokines (TNF-, IL-6), inducible nitric oxide synthase (iNOS) and important transcriptional factors (NF-E2related factor 2NRF2 and silent mating type information regulation 2 homolog) 1 C SIRT-1) were quantified by real-time
polymerase chain reaction.We observed a reduction in the expression of the folling enzimes: SOD in MTN (0,61) and MTD
(0,64) compared with control group (1,07 , p <0.05).
Conclusions:
We suggest that MT can minimize the aging process and protect the mouse lung through its antioxidant and anti-inflammatory
properties.
Keywords: aging, mate tea, stress oxidative
Financial Support: Faperj, CNPq, Mate Leao, Capes.
Resumo:26-229
EVALUATION OF ACUTE TOXICITY (LD 50) AND OSMOTIC FRAGILITY OF CRUDE AQUEOUS EXTRACT OF
TURNERA SUBULATA.
Costa, R. C. D. 1; Silva, T. R. P. M. 1; Barreto, T. L. P. 1; Silva, N. M. B. 1; Brito, M. M. V. 1; Gouveia, A.

L. A. 1; Magnata, S. S. L. P. 2; Catanho, M. T. J. A. 1
1
Depto de Biofsica e Radiobiologia/Universidade Federal, UFPE
2
Centro Acadmico de Vitria (Ncleo de Cincias Biolgicas), CAV-UFPE
Objectives:
.Despite the investigations and scientific studies on plants with medicinal use, not much is known about the active ingredients and
the extraordinary healing qualities of many plant species. The species Turnera, Turneraceae family, are known in the Brazilian
Northeast by the popular name of "chanana, " whose roots, sold in street markets in the region, are used in folk medicine to treat
amenorrhea, dysmenorrhea and as abortifacient. Other species, such as Turnera diffusa Willd. and Turnera ulmifolia L., are used
primarily as an aphrodisiac, abortifacient, expectorant, to treat gastric ulcers and diabetes. Turnera subulata is used for
amenorrhea in the form of tea. Evaluation of Acute Toxicity (LD 50) and Osmotic Fragility of crude aqueous extract of Turnera
subulata

We used 20g leaves of T. subulata, they were dried and crushed to obtain the crude aqueous extract by infusion method and
lyophilized. For the toxicity test were used Wistar rats weighing on average 30g, the lyophilized aqueous extract of T. subulata
was dissolved in distilled water and administered orally (gavage), a group of 15 animals. The animals were divided into 3 (three)
groups of 5 (five) animals each. Group I received a dose of 250mg/kg, 100mg/kg and II received the third received 500mg/kg
body weight. To test osmotic fragility was used the blood of adult Wistar rats of both sexes, weighing between 200 and 250
grams. Heparinized blood (500L) was incubated for 1 hour in 500L of crude extract in different concentrations (0%, 50 and
100% volume / volume). Subsequently, aliquots of 50mL were subjected to a gradient of NaCl (0%, 0.1%, 0.25%, 0.4%, 0.7%
and 0.9%) for 1 hour. The percentage of osmotic fragility and determined by optical density (OD / 545nm) samples. The
morphology of RBC was evaluated by estiraos blood, dried at room temperature, fixed, stained with Giemsa and visualized by
optical microscopy (x100) of Leica. Results: In the determination of toxicity within 48 hours, no lethality was observed. The
evaluation of erythrocyte osmotic fragility was studied subjected to different concentrations of crude extract of T.subulata
showed a 40% change in concentration of 0% NaCl% when compared to control, demonstrating that the crude extract of T.
subulata decreased osmotic fragility. The analysis of the morphology of red cells corroborated the results of osmotic fragility,
showing that at concentrations of 0% and 0.1% the cells typically showed hemolisadas.
Conclusions:
In the study on acute toxicity (DL 50) was not observed lethality in any of the doses administered. The effect of the extract of T.
subulata on the erythrocyte membrane osmotic fragility was decreased by increasing the resistance.
Keywords: Toxicity, osmotic fragility, extract, Turnera subulata
Financial Support: CNPq-Pibic.
Resumo:26-230
INVOLVEMENT OF POTASSIUM CHANNELS IN VASORELAXANT EFFECT INDUCED BY VALERIANA
PRIONOPHYLLA STANDL. IN MESENTERIC ARTERY
Ramos, M. 1; Arajo, J. P. B. 1; Maciel, P. M. P. 1; Queiroz, T. M. 1; Cavalcante, K. V. M. 2; Cechinel Filho,

V. 3; Silva, D. F. 4
1
Ps-graduao em Produtos Naturais e Sintticos Bioativos, UFPB
4
Depto. de Biorregulao/ICS, UFBA
3
Ncleo de Investigaes Qumico-Farmacuticas/CCS, UNIVALI
2
Depto. de Fisioterapia/NEST, UFRN
Objectives:
The aim of this study was to investigate the mechanisms underlying the vasorelaxant response induced by metanolic extract of
roots (VPR) and metanolic extract of leaves from Valeriana prionophylla Standl. (VPF) in isolated rat mesenteric artery.
Isolated rat superior mesenteric artery rings (1-2 mm) were suspended by cotton threads for isometric tension recordings in a
Tyrodes solution at 37C, gassed with a 95% O2 and 5% CO2, under a resting tension of 0.75g. In phenylephrine (Phe, 1M)precontracted mesenteric artery rings, VPR (0,01 300 g/mL) caused a concentration-dependent relaxation (Maximum
Response (MR) = 75.43 4.08%, EC50 = 5.96 (3.8 9.2) g/mL, n=6). After removal of the vascular endothelium, VPR caused
a concentration-dependent rightward shift of the response curves, despite no changes have been observed in the MR (MR = 70.33
4.58%; EC50 = 39.63 (27.2 57.6) g/mL; P < 0.05; n=5). However, VPF (0,01 300 g/mL) induced vasorelaxant effect in
mesenteric artery rings , but this response was less than that obtained by VPR. Based on the preliminary results, the subsequent
experiments were performed to investigate the endothelium-independent relaxation induced by VPR. In preparations denudedendothelium pre-incubated with Tyrodes modified solution, KCl (80 mM) and KCl (20 mM), the vasorelaxant effect of VPR was
attenuated (MR = 40.29 8.22%, MR = 50.48 6.00 % respectively, n=5). To investigate the involvement of K+ channels, the
preparations were pre-treated with tetraethylammonium (TEA, 3 mM), a non-selective K+ channel blocker, or with TEA (1 mM),
a large-conductance Ca2+-activated K+ channels selective blocker (BKCa). In both preparations the vasorelaxant activity of VPR
were significantly attenuated (MR = 46.79 6.12%, n = 8; MR = 49.33 6.39%, n=7, respectively). In contrast, neither
glibenclamide (10M), barium chloride (30M) nor 4-aminopyridine (1mM) affected VPR-induced relaxation.
Conclusions:
Taken together, these results demonstrate that endothelium-independent relaxation induced by VPR involves K+ channels
activation, probably BKCa channels, in the rat superior mesenteric artery.
Keywords: Natural products, Valeriana prionophylla Standl. , Vasodilatation, Mesenteric artery, Potassium Channel
Resumo:26-231
IN VITRO ANTIMICROBIAL ACTIVITY OF ETHANOL EXTRACT OF THE LEAF ON EVOLVULUS ELEGANS
STAPHYLOCOCCUS AUREUS. 1LEO,A.D.,2LUCENA,K.L.,3ANTUNES,PS.,4DUARTEM.C.,5QUEIROGA,C.S.,6
ARAJO,J.I.R.,7SILVA,K.H.P.O.,8FARIAS, R.L.G.P 9NASCIMENTO,J.S.,1DEPARTMENT OF PHYSIOLOGY AND
PATHOLOGY, UNIVERSIDADE FEDERAL DA PARABA, 58051-900, JOAO PESSOA, BRAZIL. 2LABORATRIO
OF PHARMACEUTICAL TECHNOLOGY. FEDERAL UNIVERSITY OF PARABA, CIDADE UNIVERSITRIA,
58051-970, JOO PESSOA, BRAZIL.
Leo,a. D. , A. D. L. . . 1,1,7,1; Lucena,k. L. , K. L. L. . 1,1; Antunes,p. S. , P. A. . . 1,1,1,1; Duarte,m. C. , M. D.

1
; Queiroga,c. S. , C. S. Q. 1; Araujo,j. I. R, J. I. R. D. A. . 1; Silva, K. H. P. O. D. 1; Farias,r. L. G. P1;
Nascimento,j. C. , J. . S. N. . . 1,1
1
Universidade Federal da Paraiba , UFPB
2
fisiologia e patologia, UFPB
3
Fisiologia e patologia, UFPB
4
5
FISIOLOGIA E PATOLOGIA, UFPB
6
7
Objectives:
Evolvulus elegans has no popular name, belongs to the family Convolvulaceae. In literature, found no study on its antimicrobial
activity. Given the above, the present study was to evaluate the in vitro antimicrobial activity of ethanol extract of leaves of
E.elegans on Staphylococcus aureus and to determine their minimum inhibitory concentration (MIC).
To perform the microbiological tests, were selected isolates of S. aureus of clinical origin. In sterile Petri dishes was seeded the
suspension of the microorganism (containing 1.5 x108UFC/mL based Mac Farland scale) with the aid of swabs sterilized. Next,
we carried out drilling of wells with a capacity of 50L using sterile tubes, which were deposited in aliquots of 50mL of the
extract, at concentrations of 0 - 25 - 50 - 100%, in triplicate. The antibiogram showed that the extract of E. elegans has not
formed inhibition zonesagainst three strains of S. aureus, where the growth of bacteria occurred in 100% in the presence of the
extract.
Conclusions:
The ethanol extract of leaves of E. eleganns not interfere with the growth of three strainsof Staphylococcus aureus in vitro, and
this is not a statement promising to control these microorganisms.
Keywords: antimicrobial activity, Convolvulaceae, Evolvulus elegans
Financial Support: UFPB
Resumo:26-232
EVALUATION OF TOXICITY IN VITRO OF ESSENTIAL OIL OF XYLOPIA LANGSDORFFIANA FRUITS
Sousa, T. K. G. ; Moura, A. P. G. ; Pita, J. C. L. R. ; Moreira, M. M. B. ; Duarte, M. C. ; Tavares, J. F. ;

Silva, M. S. ; Diniz, M. F. F. M. ; Pessoa, H. L. ; Castello-branco, M. V. S. C.
Objectives:
Xylopia langsdorffiana is a tree popularly known as pimenteira-da-terra. The phytochemical study of essential oil of X.
langsdorffiana fruits (O.E.X.) characterized substances with important biological activities as -pinene (37.73 %) and limonene
(31.42 %). To evaluate the cytotoxicity O.E.X. we used two assays: hemolytic activity and brine shrimp test. The erythrocyte
membrane is a delicate structure that can be significantly altered by interactions with drugs. The in vitro hemolysis test is used as
a screening method for estimating the toxicity of substances which can induce in vivo erythrocytes damage (Biochem. Pharmacol.
41:133, 1991). The significant correlation between the bioassay with Artemia salina and in vitro growth inhibition of cancer cell
lines shows that this method may be a useful tool for screening in the investigation of antitumor drugs (Phytochem. Anal. 2:107,
1991).
Swiss mice blood samples were suspended in Phosphate Buffer Solution (PBS) to make 1 % solution of erythrocytes (Comp.
Biochem. and Physiol. 150:85, 2009). Then, O.E.X. (0-300 g/mL) in DMSO (5%) was added to the suspension, which was
incubated on a homogenizer for 60 min and then centrifuged. The absorbance of supernatants at 540 nm was read to determine
the concentration that produces 50 % hemolysis (HC50). Positive control and negative control were also used with 1 % Triton X100 and PBS alone, respectively. To brine shrimp test, we used larvae of Artemia salina L., using the median lethal concentration
(LC50) as the parameter of biological activity. Eggs of A. salina were incubated in artificial sea water and light during 24 h for
obtain the larvae. O.E.X. (0-1000 g/mL) was added in tubes containing 10 nauplii. The set was incubated at artificial light for 24
h and then the survivors larvae were counted to determine the LC50 (Phytom. 8:395, 2001). The HC50 and LC50 values and their
95 % confidence intervals were obtained by nonlinear regression, using GraphPad Prism. The percentage of hemolysis increased
concentration dependent manner after treatment with the O.E.X. that produced 100% hemolysis at concentrations above 300
g/mL. The HC50 value obtained was 68,1 (63,6 - 73,0) g/mL. The LC50 value obtained in brine shrimp test was 459,0 (450,6 467,6) g/mL. Considering the criterion of activity being LC50 values of A. salina. Tests for assess the antitumor activity are
being conducted for comparison with data of cytotoxicity in erythrocytes, since this cell type is one of the most affected during
cancer treatment.
Conclusions:
Therefore, we conclude that the O.E.X. showed to be bioactive against A. salina and showed moderate toxicity against
erythrocytes of mice.
Keywords: Artemia salina, Cytotoxicity in erythrocytes, Essential oil , Xylopia langsdorffiana
Resumo:26-233
EVALUATION OF CYTOTOXICITY AND ACTIVITY OF ANTI-TRYPANOSOMA CRUZI COUMARIN
COMPOUNDS
Oyafuso, L. T. ; Pereira, R. M. S. ; Santos, M. R. M.

Lab of Cellular Biology/ Bandeirante University of So Paulo, UNIBAN
Objectives:
Although known since 1909, when it was described by public health physician Carlos Chagas, Chagas disease, also called
American trypanosomiasis, still has great public health importance in Brazil, considered one of the four major endemic diseases.
Due to the high toxicity of the drug (benznidazole) and its low efficacy in chronic disease, currently there is exists great interest
in research with the aim of finding new therapies for treatment. Considering the existing biodiversity in Brazilian fauna and flora
and their potential as a source of bioactive molecules to be studied and explored, our research group grounded in previous studies
of the characteristics and activities of coumarin analyzed this molecule and its derivatives obtaining promising effects in the
treatment of neglected diseases.
In experiments in vitro we used benznidazole, coumarin and three bioactive compounds derived from coumarin: nitrocoumarin;
3,4 hydroxy-nitrocoumarin and 3 carboxy-8-nitrocoumarin, diluted as its solubility in water or dimethyl sulfoxide. The evaluation
of the cytotoxicity was performed by reducing the bromide 3-(4,5-DimethylTiazol-2-yl)-2,5-difeniltetrazlio (MTT) in VERO
cells (African green monkey kidney cells fibroblasts) distributed in 96-well plates with 2x104 cellswell in 200 L of RPMI
(Roswell Park Memorial Institute - 1640), maintained in an incubator at 37 degrees in 5% CO2, with RPMI or different
concentrations of compounds ranging from 25 to 500 gmL for 24 hours. The analysis of the different concentrations of the
compounds was done by reading the absorbance values of triplicates in a 570nm spectrophotometer. The data were converted to
percentage of cell viability to determine the IC50 concentration through the equation line, where benznidazole IC50 = 189,3 g;
coumarin IC50 = 230,5 g; nitrocoumarin IC50 = 182,3 g, 3,4 hydroxy-nitrocoumarin IC50 = 262,5 g and 3 carboxy-8nitrocoumarin IC50 = 407,9 g. The assessment of antiparasitic activity was conducted in VERO cells infected with
Trypanosoma cruzi CL strain and after establishment of infection 106 cells were distributed in 24-well plates containing
coverslips. The tests were performed in triplicates and the results were determined by counting total of 200 infected and
uninfected cells, and in the infected cells were also counted the number of intracellular amastigotes. After completion of the
counts, with the average of triplicates it was calculated the percentage of infected cells and the concentrations that have the
lowest percentage of infection were benznidazole 5 g (7%), 10 g coumarin (11,71%), nitrocoumarin 50 g (9,66%), 3,4
hydroxy-nitrocoumarin 50 g (7,74%) and 3carboxy-8-nitrocoumarin 20 g (10%).
Conclusions:
We can conclude that coumarin and its derivatives have good antiparasitic activity in Chagas disease and they suggest that the
continuity of work in the new tests in vivo.
Keywords: CYTOTOXICITY, COUMARIN, CHAGAS DISEASES
Financial Support: UNIBAN
Resumo:26-234
EVALUATION OF ANTIPLATELET ACTIVITY OF MONOTERPENE PULEGONE
Flix, J. A. 1; Magalhes, R. A. 1; Leal, L. K. A. M. 1; Bastos, M. V. R. 2; Viana, G. D. A. 1; Viana, G. S. D.

B. 2; Flix, F. H. C. 3; Fontenele, J. B. 1
1
Farmcia/Faculdade de Farmcia,Odontologia e Enfermagem, UFC
2
3
Hospital Infantil Albert Sabin, HIAS
Objectives:
To evaluate the antiplatelet activity of Pulegone in human PRP.
Platelet aggregation was measured according to the method of Born and Cross (J Physiol. 163; 178-195, 1963). Briefly, platelet
aggregation was induced at 37C in the aggregometer, with stirring at 1000 rpm, by addition of ADP (4M) as agonist. The
resulting aggregation, measured as the change in light transmission, was recorded for 5 min and presented as percent aggregation
related to control (100%). Pulegone oil (PUL) was emulsified with Tween 80 (660 g mL1, control) and concentrations of 110,
220, 440 and 660 g mL1 were used. PUL 220, 440 and 660 g mL1 reduced significantly (p
Conclusions:
Direct stimulation of platelets by ADP results in shape change, reversible aggregation at physiological concentrations of calcium,
and finally, desensitization. Transduction of the ADP signal involves a transient rise in free cytoplasmic calcium, due to the
mobilization of internal stores and secondary store-mediated influx, and a concomitant inhibition of adenylyl cyclase activity. It
is possible that the antiplatelet effect of pulegone is due to blocking of a step of ADP-induced platelet stimulation. Statistical
analysis: All values are expressed as mean SD. Differences between the sample-treated and control groups were submitted to
analysis of variance (ANOVA) followed by Tukey-Newman-Keuls test for multiple comparisons (p < 0.05) was considered
significant.
Keywords: ANTIPLATELET, PULEGONE, ACTIVITY
Financial Support: FUNCAP
Resumo:27-049
DECISION MAKING AND EMOTION
Brizante, J. G. ; Baldo, M. V. C.
Fisiologia e Biofsica / Instituto de Cincias Biomdicas, ICB-USP
Objectives:
It has been observed that normal individuals tend to avoid risky outcomes in tasks of decision-making, usually preferring to
choose a source of smaller gains accompanied by smaller losses instead of choosing a source of large gains accompanied by also
large losses (Games and Econ Behav 52: 336, 2005). Our objective was to verify whether emotionally valenced pictures can
interfere in this process in at least two ways: a bias for avoiding (or taking) more risk; and an increase (or decrease) in the time
spent to make decisions depending on the valence of the image presented just prior the subjects choice from two differently
rewarding sources.
Methods : Thirty-five volunteers (16 women, mean age 22,9 years) were presented with two decks of cards on the computer
screen. Their task was to choose cards from each deck in a way they could get as much money as possible. One of the decks gave
rewards of R$0,16 and penalties of -R$0,16 (Low Risk: LR) and the other one offered rewards of R$0,32 and penalties of R$0,80 (High Risk: HR). The net gain of the LR and the HR were R$250,00 and -R$250,00, respectively. The presentation of the
pair of decks was preceded by a neutral, positive or negative picture (IAPS - International Affective Picture System) at the center
of the screen for either 10ms or 200ms, depending on the experimental condition. The experiment totalized four blocks: 1)
Neutral; 2) Negative (Positive); 3) Neutral; and 4) Positive (Negative) the presentation order of positive and negative pictures
was randomized. The subjects started the test with a positive balance of R$20,00. Results There was no influence of time of
exposure either on the risk-taking behavior or on the time spent to make decisions (p>0.01). Besides, there was no influence of
valence on the willing to take risk, but a significant effect of valence was found in the time spent to make decisions, which was
higher in the Negative block than in the Positive block (p=0.043).
Conclusions:
The emotional valence of the pictures presented didnt influence the strategy the subjects developed in the test. Considering the
risk taking, it seems that the motivation to gain money was refractory to any other affective interference in the decision making
process. Across trials, subjects tended to be conservative, with the HR deck being selected in only 30% of the trials. However,
there was a significant valence effect on the decision time. In trials with negative valence images preceding a decision, subjects
spent more time to reach their choices than they did in trials with positive valence previous images.
Keywords: decision-making, emotion, economic decision
Financial Support: Coordenao de Aperfeioamento de Pessoal de Nvel Superior (CAPES).
Resumo:27-050
REDUCED ORBITOFRONTAL CORTEX THICKNESS IN FEMALE PATIENTS WITH BORDERLINE
PERSONALITY DISORDER.
Araujo, T. B. D. 1,2,3; Araujo, G. M. F. D. 1,2,3; Sato, J. R. 4,2; Jackwoski, A. P. 1,2

1
Psiquiatria Unifesp , Unifesp
2
3
Laboratrio Interdisciplinar de Neurocincias Clnicas, Linc

Ambulatrio de Transtorno de personalidade , AMBORDER
4
Centro de Matemtica, Computao e Cognio, UFABC
Objectives:
Borderline personality disorder (BPD) is a devastating condition that affects 1% to 2% of the population and causes an intense
disruption of patients lives and relationships. BPD patients often exhibit emotional instability, impulsive behaviors (selfmutilation, substance abuse, sexual promiscuity and binge eating), unsteadiness, rapid mood changes and a propensity toward
intense negative emotional states like anger, anxiety and dysphoria. Proper understanding of the neurobiological bases of BPD
could contribute to clinical management of these patients and to provide the basis for earlier and effective interventions. Recent
evidence from neuroimaging studies have suggested that changes in frontal gray matter could help to understand the
pathophysiology of BPD. Once there is a scarse of structural and functional neuroimaging studies in patients with BPD, we
assessed the volume and thickness of orbitofrontal cortex (OFC), an important subdivision of prefrontal cortex and involved in
emotional processing, aiming to verify possible OFC volume and thickness reductions in BPD patients compared to healthy
controls.
Methods: Twenty-five female BPD patients treated as outpatients in a tertiary referral center (Department of Psychiatry,
Universidade Federal de So Paulo, So Paulo, Brazil) were included in the study. After approval by the Ethics Committee
(1246/10), they underwent a psychiatric evaluation through SCID I and II. The clinical global impression (CGI) scale was also
applied to evaluate the severity of BPD. The control group was formed by 25 female healthy controls from the community. MRI
examination of the brain was obtained from all subjects using a 1.5 T (Magnetom Sonata [Maestro Class] Siemens AG, Medical
Solutions, Erlangen, Germany) using an eight-channel head coil. To minimize variation, the subjects were positioned by the same
investigator using the orbito-meatal line as landmark. Structural images were processed using the recon-all pipeline of Freesurfer
package, a software used to study the anatomy of cortical and subcortical. Results: The mean age was (32,79,09 years) for BPD
groups and (32,27,07 years) for controls. Groups were matched for age and handedness. BPD patients showed a significant
reduction in medial and lateral regions in left (p = 0.039 and 0.016, respectively) and right (p = 0.042 and 0.026) OFC
hemispheres compared with healthy individuals.
Conclusions:
The present study showed a significant reduction of OFC thickness in a group of female BPD patients compared with a healthy
control group paired by age, gender and handedness. In accordance with previous structural neuroimaging studies involving
patients with BPD, the present study added new evidence of OFC involvement BPD through Freesurfer, a method of structural
analysis that can contribute to this discussion and offering different simultaneous volumetric and geometric parameters. Although
these data support the hypothesis of OFC involvement in the pathophysiology of BPD symptoms, more neuroimaging studies in
BPD are needed in order to understand its biological underpinnings.
Keywords: borderline personality disorder, orbitofrontal cortex, structural neuroimaging
Financial Support: This work was supported by CNPq from Brazil.
Resumo:27-051
EVALUATION OF CHANGES PROMOTED BY COCAINE ALONE AND IN COMBINATION WITH SILDENAFIL
ON THE PROCESS OF FORMATION OF MEMORY IN RATS
Costa, J. M. ; Cruz, G. M. P. ; Carvalho, A. B. ; Leite, T. A. ; Alcntara, G. F. T. ; Viana, G. S. B. ; Lima, I.

S. P.
Faculdade de Medicina Estcio de Juazeiro do Norte, FMJ
Objectives:
Cocaine remains as one of the most widely used drugs of abuse in several countries and its use results in large acute and longterm losses for the users. A recent association that has been observed is the combination of cocaine with sildenafil, a drug for
impotence. This study aimed to analyze memory changes by the use of cocaine alone and in combination with sildenafil.
Male Wistar rats (2 months old, 200-250g, n=8 animals) were divided into four groups: cocaine (Coc 20mg/kg, intraperitoneal
(i.p)), sildenafil (Sil 5mg/kg, i.p) and their association (Coc + Sil 5mg/kg, 20mg/kg, i.p). The control group received distilled
water. The animals were treated for 5 days. After 24 hours of the last treatment, animals were subjected to behavioral tests. To
assess memory, it was used the method of the water maze (WM). In WM (black circular pool with water at 25C, located in a
room with visual cues on the walls), each animal was released from a position to find a platform submerged 2cm, with 54s to find
it and remaining there for 10s. The process was repeated five times the training phase. The training was repeated after 24h, and
the test performed after 48h, where each animal was placed only once in the pool. It was also made a passive avoidance test to
analyze memory. The animals were accustomed in a device that consists of an acrylic box (48 x 22 x 22cm) divided into a light
and a dark compartment. The light compartment had a wire mesh floor, and was illuminated with a 24V-5W bayonet lamp. The
dark compartment was made of black acrylic with a floor connected to a shocker to provide a footschock. The two compartments
were separated by a sliding door. The rat was initially placed in the light compartment for 1 min (habituation). After that, the
animal was removed and fifteen minutes after it was put again in the bright compartment to evaluate short-term memory. After 24
hours the experimental procedure was repeated to evaluate the long-term memory. Results were expressed as means SEM
(ANOVA and Student-Newman-Keuls; p< 0,05). The results showed that in the water maze cocaine caused a significant
reduction in memory. (Coc 20mg/kg: 35,5 3,5 s; Control: 20,2 3,1 s), but it was not altered by sildenafil in the group treated
with the association (Coc 20mg/kg + Sil 5mg/kg: 37,4 2,8 s). In the passive avoidance test, we also observed that after 24h
cocaine significantly modified building memory process (Coc 20mg/kg: 48,5 20,1 s; Control: 237,0 30,9 s), an effect not
modified by sildenafil in the group treated with the combination (Coc 20mg/kg + Sil 5mg/kg: 35,3 15,5 s).
Conclusions:
Our results demonstrate that treatment with cocaine reduces the process of memory formation. Sildenafil, which presents some
studies on affirmative action in the process of memory formation, was unable to reverse the actions made by cocaine. Further
studies with larger numbers of doses are thus needed to evaluate the real impact of this association used by some cocaine users.
Keywords: Cocaine, Sildenafil, Memory, Cocaine and Sildenafil
Financial Support: Faculdade de Medicina Estcio de Juazeiro do Norte - CE
Resumo:27-052
ENVIRONMENTAL ENRICHMENT PROMOTES BEHAVIORAL CHANGES IN ALBINO SWISS MICE IN
OBJECT RECOGNITION TASK
Viola, G. G. 1; Binder, L. 2; Muccini, E. 2; Martins, W. C. 1; de Oliveira, P. A. 3; Prediger, R. D. 3; Tasca, C.

I. 1,2,3
3
PPG Neurocincias , UFSC

Departamento de Bioqumica, UFSC
Objectives:
Enrichment environmental (EE) protocol is an experimental model that allows studies of neuroplasticity in laboratory animals. In
rodents, EE promote increase in physical activity, learning experiences and social interactions. EE exacerbates the behavioral
differences among the laboratory species and/or strains and makes the behavioral response more similar to behavioral responses
presented by wild animals. The object recognition task (ORT) deals with the natural motivation of the humans, rats and mice
approaching and exploring novel objects rather than familiar objects (as new/unfamiliar preference), an innate instinct that drives
animals to learn about their environment (discrimination ratio). In this work, the objects were not known and not have ethological
significance for mice. The aim of this study is evaluate the changes in behavior of albino swiss mice maintained in EE in the
ORT.
Swiss mice (n=20) were weaned for 21 days and assigned randomly to standard or enriched housing immediately after weaning
for 60 days. Standard housing consisted of a 42 cm x 32 cm x 17 cm acrylic box with sawdust containing groups of 10 mice.
Enriched housing consisted same acrylic box connected to a 28 cm x 21 cm x 50 cm three-story metal cage with sawdust, housing
10 mice at a time. The enrichment housing apparatus contained three running wheels and a variety of objects, including tunnels;
hiding places and nesting material, where the mice were keep out of luminosity, the natural behavior of wild mice. After eight
weeks the animals were habituated to the experimental room for 60 minutes at dim light conditions. A light bulb was switched on
during the experimental sessions. The time spent exploring in both objects decrease in the EE group and decrease in the different
sessions. The discrimination ratio is similar between control and EE in training session (0.5038 0.021 vs 0.4759 0.052). Both
group increase the discrimination ratio in test 24h (0.6022 0.0588 vs 0.5692 0.1038) and test 48h (0.5948 0.069 vs 0.5946
0.088) indicating that the animals learned about the environment.
Conclusions:
The EE groups spent less time exploring both objects which indicates a more rapid exploration, minimizing associated costs
and/or indicate reduction of motivation, curiosity and/or interest for objects, probably because these animals previously
experienced more stimulating environmental conditions (learning, social and physical), which make the novelties not so
appealing. The discrimination ratio was similar in control and EE groups and increased in tests sessions compared to training
sessions, these results indicate a more propitious behavior for species survival, including rapid exploration and knowledge about
the environment.
Keywords: ethology, ENVIRONMENTAL ENRICHMENT, OBJECT RECOGNITION TASK, behavior
Financial Support: CNPq, CAPES, INCT, FAPESC
Resumo:27-053
EFFECT OF THE AD LIBITUM INGESTION OF ERVA-MATE (ILEX PARAGUARIENSIS) TEA ON BEHAVIOUR
IN THE OPEN FIELD AND ELEVATED PLUS MAZE, IN MALE WISTAR RATS AT 240 DAYS OF AGE.
Nakatani, M. ; Cantelli, K. R. ; Fronza, J. L. ; Padoin, M. J.

CCBS/Universidade Estadual do Oeste do Paran, UNIOESTE
Objectives:
Erva Mate(Ilex paraguariensis) is a plant widely used in the southern region of Brazil to make chimarro, terer and mate tea. It
contains various compounds that can influence metabolism. This study examined the effect of the ad libitum ingestion of erva
mate tea on selected behaviours of Wistar rats at 240 days of age.
Control Group (CG; n=12) received water; Ingestion group (IG; n=12) received erva mate tea. The tea was prepared daily, with
the erva mate(g) being added to boiling distilled water (mL) at a ratio of 20:1 (50g of Ilex paraguariensis to 1000 mL of water).
The erva mate was left to infuse in the boiling water for 5 minutes. The tea was filtered, cooled and given to the IG, for 60 days,
from 240 days of age. The behaviours analysed in the open field were:locomotion in the periphery, remaning stationary,
locomotion in the center, rearing and grooming. In the elevated plus maze the following were assessed: time spent on open arm,
time spent on closed arm, open arm head dipping and closed arm head dipping. Each rat was filmed for 5 minutes in the dark
period, witch is when the animal is most active. The parameters of latency, frequency and duration were examined in all animals.
There were no significant differences in the behaviours assessed in the open field as shown by the frequencies of locomotion
(CG=6.20.7; IG=4.80.6, rearing(CG=13.61.9; IG=12.32) and grooming (CG=7.41.2; IG=6.40.8). Similarly, there were
no differences in the behaviours assessed in the elevated plus maze; time spent on closed arms (CG=183.926.3; IG=208.917),
time spent on open arms (CG=112.526.1; IG=87.516.8), closed arms head dipping (CG=39.88.1; IG=36.36.5), and open
arms head dipping (CG=38.110; IG=24.55.4).
Conclusions:
The results indicate that the ad libitum ingestion of Ilex paraguariensis tea, by male Wistar rats aged 240 days, has no significant
influence on the animals behavior in either the open field or elevated plus maze.
Keywords: behavior in the open field, behavior in the elevated plus maze, ingestion of erva-mate
Financial Support: Unioeste
Resumo:27-054
DISTINCT SUSCEPTIBILITY TO INCORPORATE INFORMATION IN DIFFERENT AGES
Crestani, A. P. ; de Oliveira Alvares, L. ; Santana, F. ; Cassini, L. ; Huabrich, J. ; Sierra, R. O; Quillfeldt, J.

A.
Depto Biofsica - Inst.Biocincias - U.F. Rio Grande do Sul , UFRGS
Objectives:
The retrieval or reactivation of a memory places it into a labile state, requiring a process of reconsolidation to restabilize it. This
retrieval-induced plasticity is a potential mechanism for the modification of the existing memory. Since experimentally False
Memories (i.e. memories drastically changed or even constructed) in humans are created when memory are reactivated, we have
analyzed the susceptibility to incorporate information upon an existing memory in different ages during retrieval.
Rats were trained in a contextual fear conditioning task in the context A (2x0.7mA footshock- day 1), reactivated in a context AB
(90 sec day 3) and tested in context B (day 5). The control group has performed the same procedure except by skipping the
reactivation session. Hypothesis: the reactivated group would update the context during the reactivation by associating the
context B with the footshock through the hybrid context (AB), whereas the control group would not. Adults animals (3 months)
that was not reactivated (17,74[12,91] N=11), (always represented by mean [S.E.M.]) showed less freezing when was tested in a
new context (B) compared with reactivated group (50,30[4,95] N=15), indicating that the reactivated group was susceptible to
incorporate information (Independent t-test p
Conclusions:
These results suggest that information can be incorporated in memory during retrieval in order to change this trace memory in
adults. However, this pattern wasnt observed, neither in young, nor in old rats, suggesting that their memories are context
unspecific. These results could be explained considering hippocampus immaturity in the younger rats and an age-related
cognitive deficit in older animals, since in either case, the contextual discrimination would be modified.
Keywords: age, memory, reactivation, susceptibility to incorporate infomation
Financial Support: CAPES, CNPq, FINEP, FAPERGS, IFS
Resumo:27-055
PRENATAL STRESS BY LIGHT/DARK CYCLE INVERSION ALTERS COGNITIVE AND EMOTIONAL
BEHAVIORS OF FEMALE OFFSPRING IN ADULTHOOD
dos Santos, E. L. 1; Bido, A. F. 1; Souza, A. P. 1; Santos, E. L. 1; de Picoli Souza, K. 1

1
Faculty of Biological and Environmental Sciences , UFGD
2
Postgraduate Program in Health Sciences, UFGD
Objectives:
The nervous system is continually shaped by the animal's interaction with the environment throughout life, however, pregnancy is
a period in which this neural plasticity is intense, which makes it more vulnerable to such variations. In this study we investigated
the effects of prenatal stress by periodic inversion of the light/dark cycle (L/Dc) during pregnancy on feeding, cognitive and
emotional behaviors of the female offspring in adulthood.
Behavioral assessment: (1) Feeding - Food (g) and water (ml) intake and body mass evolution (g); (2) related to Anxiety: Open
Field - OF (n of squares crossed, freezing time, n of responses of grooming and rearing, n of feces defecated and urine) and
Elevated Plus Maze - EPM (n of entries and time spent in open and closed arms, central region, freezing, n of responses of
grooming/ rearing/ head-dipping/feces defecated and urine), and (3) Memory and learning - Morris Water Maze - MWM (latency
time to find the platform). Female Wistar rats of 90 days mated and at the first day of pregnancy checked by vaginal swab, were
separated into two groups: Control Group (CG, n = 5) animals maintained under standard L/Dc (12 h in light and 12 h in
darkness) during pregnancy and Prenatal Stress Group (PSG, n = 9) animals submitted to periodic inversion of the L/Dc each
three days during pregnancy period. In the first postnatal day of the offspring, the genitors were evaluated in OF and EPM to
investigate their stress level. The female offspring of CG (n = 8) and PSG (n = 16) were analyzed between PN21 and 105 for
feeding behavior, PN65 and 70 for behaviors related to anxiety and PN100 and 105 for memory and learning. Data were
expressed as mean SEM and submitted to statistical analysis by Student t test. The differences were considered significant when
p
Conclusions:
Our results show that the inversion of L/Dc during pregnancy is a stressful event for genitors with long time consequences for
offspring emotional and cognitive behaviors in adulthood, resulting in increased anxiety and memory and learning deficit.
Keywords: Animal model, Anxiety, Behavior programing, Learning, Memory
Financial Support: UFGD, FUNDECT and CNPQ.
Resumo:27-056
AN INNOVATIVE INTERVENTION TO ASSIST MOTOR AND COGNITIVE REHABILITATION OF STROKE
PATIENTS
Moura, D. M. S. 1; Souza, B. C. 2; Coutinho, E. A. G. 2; Cavalcanti, A. 3; Braga, A. 4; Grego, B. H. C. 1;

Alchieri, J. C. 3; Santos, S. R. D. 2; Campos, T. F. 4; Junior, A. P. 1
1
Programa de Ps Graduao em Psicobiologia, UFRN
2
Departamento de Informtica e Matemtica Aplicada, UFRN
3
Departamento de Psicologia, UFRN
4
Departamento de Fisioterapia, UFRN
Objectives:
Stroke is the worlds leading cause of cognitive and motor disabilities in adults and frequently leads to hemiparesis or partial
paralysis of one side of the body. This hemiparesis can profoundly impair activities of daily living (ADLs). The cognitve sequela
caused by stroke in the frontal lobe, for instance, are extremely debilitating as the physical sequelae, but are harder to diagnose
and receive less attention in rehabilitation programs. The underlying mechanisms for both spontaneous and therapy-induced
recovery are poorly understood. Preliminary evidence suggests that simply moving or passively exercising the impaired limb will
not lead to maximum recovery. For more than 50 years, mental practice with motor imagery has been used as an adjuvant
technique to the learning of motor skills and to increase athletic performance. Working memory is involved with the on-line
maintenance and active manipulation of information. It is generally conceived as a multicomponent system, which relies on a
complex network of temporoparietal and frontal areas. Improving working memory capacity leads to better performance of a
range of tasks such as problem solving and it translates in increased attentiveness in ADLs. In this project, we present a therapy
that combines elements of motor imagery and working memory in a computer game to allow the recovery of both upper limb and
cognitive function of stroke patients.
The patient played the computer game for 30 minutes, 3 days a week, during 6 weeks, while undergoing conventional physical
therapy. The computer game consisted of a set of working memory and motor imagery tasks played in an adaptive configuration.
The patient was recruited in the acute phase and was between 30 and 60 years old. The project was approved by our Universitys
ethics committee (number 064/10 CEP/UFRN) and conducted at the Hospital Universitrio Onofre Lopes (HUOL). The patient
reported that during motor imagery he could feel an urge to move his own hands. We also used several minigames aiming to
engage working memory. The patient also reported that the game improved his concentration in daily life activities. The patient
had no difficulties in understanding the rule of the game. Preliminary results with the Weschle Adult Intelligent Scale (WAIS)
showed the patient improved his score after the intervention, when compared to the baseline, particularly in processing speed and
perceptual organization. The Wisconsin Card Sorting Test (WCST) also detected an improvement in successful category
completion, with less persevarative errors when compared to the pretest condition.
Conclusions:
Our innovative therapy combining elements of motor imagery and working memory had a positive reception and improved
cognitive function in stroke patients, according to preliminary results.
Keywords: neurorehabilitation, motor imagery, stroke, cognition, computer game
Financial Support: CAPES, UFRN, PRONEX
Resumo:27-057
STUDY ACTIVITIES OF DAILY LIVING IN PATIENTS WITH PARKINSON'S DISEASE
Lucena, F. M. D. S. ; Jnior, J. R. D. S. ; Neto, E. D. S. R. ; Almeida, C. D. A. ; Campelo, C. L. D. C. ;

Lima, A. L. S. D. ; Franco, C. I. F.
Fisioterapia/ Universidade Estadual da Paraba, UEPB
Objectives:
Parkinson's Disease (PD) is an abnormal acceleration of the aging process that affects the elderly frequently, causing signs and
symptoms that interfere with functional independence and quality of life. Thus, this study aimed to analyze the Activities of Daily
Living (ADL) in patients with PD.
Methods: The sample consisted of eight patients of both sexes with a clinical diagnosis of PD, assisted at the Physical Therapy
Clinic UEPB. The instruments used were the Neurological Assessment Protocol for socio-demographic, Stage of Evolution
modified Hoehn and Yahr (EHYm) to classify the disease stage and the CIF, to assess ADL and UPDRS. Data were analyzed
using Graph Pad Prism 4.02, and values are expressed in percentage, mean and standard deviation, considering significant values
with p
Conclusions:
Based on data obtained is possible to suggest that the ADL in patients with PD are committed to writing and relevance in the
tremor, followed by self-care hygiene and dressing.
Keywords: Activities of Daily Living , CIF, Parkinson's Disease
Financial Support: PROBEX/UEPB
Resumo:27-058
INTRAHIPPOCAMPAL OKADAIC ACID ADMINISTRATION: AN ANIMAL MODEL FOR ALZHEIMER LIKE
DISEASE
Zimmer, E. R. ; Kalinine, E. ; Haas, C. B. ; Torrez, V. R. ; Muller, A. P. ; Souza, D. O. D. ; Portela, L. V.

Departamento de Bioqumica/UFRGS, PPGBIOQ/UFRGS
Objectives:
The western lifestyle and the increase in the lifespan have contributed to the increase prevalence of Alzheimers disease (AD).
This disease affects the brain and lead to a progressive cognitive and memory impairment. The pathological characterization of
AD is composed by accumulation of senile plaques, abnormal phosphorylation of tau and cerebral inflammation. The
intracerebral administration of okadaic acid (OA), a potent neurotoxin, has been much used in models of neurodegeneration. The
OA causes a selective inhibition of protein phosphatase 2A (PP2A) and can induce a hyperphosphorylation of tau an in vivo
models, glial damage and alterations in behavior, including cognition impairment. The main goal of this work was establish a
like-Alzheimer model with only a single intrahippocampal (i.h) administration of okadaic acid.
Male Wistar rats 4-5 months old (n=24) were divided in two groups: control (CO) and okadaic acid (OA). An infusion of OA
(100 ng) was made at the right hippocampus. To analyze alterations in the behavior and cognition performance we used the open
field test (locomotory and exploratory activity) and Morris water maze (spatial memory). For neurochemical effects of OA we
performed HPLC measurement of glutamate levels in cerebrospinal fluid (CSF), western blot of phospho-Tau in hippocampus
homogenate and imunohistochemistry to glial fibrillary acidic protein (GFAP) in hippocampus slices. The results showed no
differences in spontaneous locomotion (n=12 per group, p=0.064), however OA causes an impairment in the cognition
performance in Morris water maze (acquisition: n=12, *p=0.0318; retention: n=12, *p=0.0412). Further increased glutamate
levels in csf (n=6, **p= 0.003), immunocontent of phosphor-Tau (n=6, *p=0.0291) and the GFAP reactivity in hippocampus
(n=3, *p =0.0298) in OA group. The data from the water maze task were analyzed using repeated-measures analysis of variance
(ANOVA), followed by Tukeys post-hoc test and the other results were analyzed by student test. Differences between groups
were considered statistically significant if P < 0.05.
Conclusions:
The results showed that like-Alzheimer model used in our work causes behavior and neurochemical alterations similar to
demonstrated in AD patients and could be a good model to study the effect of this disease in central nervous system.
Keywords: Alzheimer Disease, Neurodegenerative Models, Okadaic Acid, Tauopathy
Financial Support: CNPq, Capes, INCTEN and Fapergs
Resumo:27-059
SPATIAL MEMORY EVALUATION OF 3XTG-AD MICE ON THE BARNES MAZE TEST
Kinoshita, D. ; Cohn, D. W. H. ; Costa-pinto, F. A. ; S-rocha, L. C.

Depto de Patologia, Faculdade de Medicina Veterinria, FMVZ-USP
Objectives:
The 3xtg-AD mice develop cerebral beta-amyloid plaques and neurofibrillary tangles on a time dependet manner, and have been
studied as an animal model of Alzheimer Disease. In this work, we sought to evaluate spatial memory on the Barnes Maze test, a
land-based spatial memory test, of 3xTg-AD mice at 4, 6 and 12 months old of age.
Thirteen 3xTg-AD mice and fourteen wild type mice were used. Animals were tested on the Barnes Maze at 4, 6 and 12 months
old. The procedures were always the same at the different ages. The Barnes Maze consisted of a circular (diameter = 90 cm), dry
maze, elevated 40 cm above the floor, with no walls, and with 12 holes (diameter = 5 cm) evenly distributed around its perimeter.
In one of the holes (target hole), an escape box was fitted, so that animals could not see it from the center of the maze. Distal
visual cues (colored geometric figures) were hanged on the walls, and the experimenter (the same for all the experiment)
remained always on the same place. Each mouse was introduced in the center of the maze inside a start box. After 10 seconds, the
start box was elevated, and the mouse was allowed to explore the maze for 3 minutes. When the mouse entered the box, the test
was ended. If not, the mouse was gently guided by the experimenter to the target hole, and forced to enter the escape box (by
gently pulling its tale on the contrary direction). After the mouse had entered the escape box, it remained there for 10 seconds,
and then returned to its home cage. Between each test, the maze was rotated, cleaned and the escape box was fitted on the same
hole (according to the visual cues), to avoid animals from using olfactory cues to solve the test. Animals were trained for 4 days,
4 trials a day. On the fifth day, the escape box was removed, and the probe trial was applied. We recorded the number of errors
(head deflections on holes that did not contain the escape box, the wrong holes), the latency (seconds) to find the target hole and
the distance (cm) travelled until finding the target hole. As at 12 months old, 3xTg-AD mice seemed to make head deflections on
same wrong holes, at this age we recorded the number of times the mouse made head deflections on an already explored wrong
hole, and named this parameter as short-term memory errors. 3x-Tg-Ad mice at all ages did not show deficits to solve the maze
(to find the target hole). Instead, shor-term memory errors were higher on these mice on some training days.
Conclusions:
Long-term memory (spatial memory) does not seen to be altered on the 3xTg-AD mice, even at 12 months old. Instead, short
term memory seems to be subtle impaired at 12 months old on these mice.
Keywords: Alzheimer disease, animal model, Barnes Maze, memory, 3xTg-AD
Resumo:27-060
THE NEURAL CORRELATES OF AFFILIATIVE AUTOBIOGRAPHICAL RECALL AND THEIR RELATIONSHIPS
TO THE RESTING DEFAULT-MODE NETWORK
Bado, P. 1; Bramati, I. E. 1; Basilio, R. 1; Paiva, F. F. 1; Garrido, G. 1; Sato, J. R. 1,2; de Oliveira Souza, R.

1,3
; Lima, D. O. 1; Zahn, R. 4; Moll, J. 1
1
Instituto D'Or de Pesquisa e Ensino, IDOR
2
Universidade Federal do ABC , UFABC
3
Universidade Federal do Estado do Rio de Janeiro, UNIRIO
4
University of Manchester, School of Psychological Sciences, NARU
Objectives:
Experimental studies in animals and studies using functional magnetic resonance imaging (fMRI) in humans have established that
a set of neural structures are essential for affiliative behaviors. A number of fMRI studies have studied brain emotional responses
using hypothetical scenarios and visual stimuli, and others using autobiographical recall. However, so far the neural correlates of
affiliative autobiographical memory remains poorly understood. This was the aim of the present study. Given the similarity of the
so-called resting state default, mode networks and networks involved in autobiographical recall, our study also investigated these
relationships.
Participants (n=15; F=7; age 26 3.5) selected autobiographical memories associated with affiliative (positive and negative) or
neutral contents. Subjects provided contextual information for each episode and chose a key word to recall each scenario during
the fMRI experiment. There were six scenarios for each category (POS, NEG and NTR). There were also a resting (REST)
condition (subject lied quietly in a scanner) and a subtract (SUBTR) condition (subjects continuously subtracted seven from a
given number). The fMRI paradigm consisted of two runs, each consisting of 9 main trials (POS, NEG and NTR), plus 3 REST
and 12 SUBTR trials. Functional images were acquired with a 3T Philips Achieva scanner with a gradient-echo, echoplanar
sequence (TR/TE =1650/22ms, Matrix/FOV =80/240mm, slice thickness=3mm, 29 slices), in addition to reference anatomical
images. Total scanning time was 1h19min. The data were analysed with SPM8. For the POS vs. NTR contrast we found
activation the hypothalamus, medial prefrontal cortex (BA32), medial frontopolar cortex (FPC, BA10/11), medial thalamus,
temporo-parietal junction (BA39) and precuneus (Prec, BA7/23). For the NEG vs. NTR contrast, we found similar results, and
additional activation of the piriform cortex (BA27) and more dorsal FPC activation. The same effect was observed when we
compared POS and NEG vs. SUBTR, with stronger effect in basal forebrain regions, including the hypothalamus. For the REST
vs. SUBTR contrast, we found activation in the typical brain default mode network, (medial prefrontal and parietal cortex, medial
and lateral temporal lobes, FPC and Prec), with great overlap with the POS and NEG vs. SUBTR comparisons. Remarkably, FPC
and Prec had stronger activation to emotional autobiographical scenarios than to REST. Finally, comparing REST vs. POS or
NEG showed robust responses in the subgenual, accumbens, and septo-hypothalamic regions (all comparison p
Conclusions:
Our results demonstrate the effects of affiliative emotions on brain activation, using autobiographical scenarios. We found that
medial fronto-parietal and hypothalamic areas were engaged by affiliative memories. We also demonstrated overlapping
activation of the brain default network at resting state. Interestingly, while this network was more activated during affiliative
autobiographical scenarios than at rest, the subgenual and septo-hypothalamic regions were more strongly activated during rest.
These findings demonstrate a dissociation between resting and autobiographical networks that was so far unknown. In subsequent
analyses, we will explore the influence of individual of emotional rating on these networks and investigate to what degree
functional connectivity is related to underlying anatomical connectivity using diffusion tensor imaging.
Keywords: autobiographical memory, emotion, fMRI, resting state
Financial Support: Instituto D'Or de Pesquisa e Ensino (IDOR); FAPERJ
Resumo:27-061
THE IMPACT OF POSITIVE AFFECT IN THREAT REACTIVITY
Arruda-sanchez, T. 1; Mocaiber, I. 3; Erthal, F. 1; Joffily, M. 1; Volchan, E. 1; Pereira, M. G. 2; Araujo, D. B.

4
; Oliveira, L. 2
1
2
Instituto Biomdico, UFF
3
Departamento Interdisciplinar, PURO-UFF
4
Instituto de Neurocincia de Natal Edmond e Lily Safra, UFRN
Objectives:
Recently, positive affect trait has been suggested as a key component for determining human variability to threat reactivity.
Furthermore, the role of attention in emotional processing is still in debate. We employed fMRI to investigate whether amygdala
responses to highly aversive pictures could be modulated by an attentional deployment task and if this modulation would be
influenced by positive affect.
Participants (n=22, 12 male; 19-37 years, mean=26,3 years, sd=) were scanned in a 1.5 T Siemens Magnetom Vision while they
viewed a display containing neutral (people) or aversive pictures (mutilated bodies) centrally presented and two peripheral bars.
They had to (a) judge the picture content as unpleasant or neutral, or (b) to judge the difference in orientation between the bars (0
or 90 orientation difference). Data analysis was performed in BrainVoyagerTM (QX 1.10) using GLM. The ROI analysis of
bilateral amygdala revealed an increased activation to aversive pictures as compared to neutral, during the picture judgment task
(p
Conclusions:
Positive affect attenuated emotional responses away from highly aversive pictures, thus facilitating attentional deployment.
Keywords: Positive Affect, Emotion, Attention Depployment, fMRI, neuroimaging
Financial Support: CAPES, PRONEX-FAPERJ, MCT-CNPq, IBN-Net.
Resumo:27-062
HAND (NON)DOMINANCE IN SIMPLE TASKS IS DISRUPTED BY 24-HOUR IMMOBILIZATION OF THE ARM.
Oliveira, M. F. 1; Helene, A. F. 2; Morya, E. 3; Sameshima, K. 1

Neurocincias e Comportamento - Inst Psicologia, NeC IPUSP
2
Fisiologia Comparativa - Inst Biocincias, IBUSP
3
Associao Alberto Santos Dummont de Apoio Pesquisa, AASDAP
1
Objectives:
Hand dominance is quite perceived in complex motor behavior, but simple tasks are usually performed similarly with both hands.
The hypothesis for this fact is that these types of tasks are accomplished by the activation of very simple and robust neural
networks in the cerebral cortex. To address the question of how sturdy these networks are, we designed an experiment where
volunteers were subsequently exposed for (1) long training in two simple motor tasks (choice reaction time, CRT, and finger
tapping, FT), (2) had the left-hand immobilized for a 24-hour period, if assigned to the immobilization group, (3) after what they
were retested on CRT and FT tasks. All volunteers were right-handed and had their left arm immobilized with cast and sling.
Volunteers signed a written informed consent term. The study was approved by the local ethical committee.
14 healthy volunteers participated in the study. Each volunteer were assigned to control (CG - n=7) or immobilized (IG - n=7)
group. Volunteers were seated in front of a computer screen positioned at eye level. In the CRT task, volunteers were exposed to
a picture of two hands with a small white square in the tip of one of the ten presented fingers, and had to respond in an ergonomic
keyboard witch finger had been marked pushing the correct key. The reaction time (ms) between the appearance of the stimulus
and the key pressing were recorded along the 800 trials that composed all the experiment, equally distributed between hand and
fingers, in 4 blocks. The median reaction time (RT) of correct trials were used for statistical analysis. In the FT task, volunteers
were instructed to press a button with their index finger repetitively, as fast as they could, for ten seconds in three blocks. The
median of the interval between consecutive presses (ms), was recorded and used for further analysis. We performed separated
ANOVAs, with the factor GROUP as between comparison, and the factors HAND, SESSION and BLOCK (RT task 1-4, FT task
1-3) as within comparisons. Tukeys post-hoc tests were performed to account for differences. Significant differences were set to
p values equal to or less than 0.5. As expected, there were no differences between hands at the session test in both groups, nor at
the retest for the CG.The ANOVA for CRT task was significant for the factor HAND (F7.534, p=0.02) and almost significant for
the factor SESSION (F4.584, p=0.058). Tukeys post-hoc tests revealed that, in the IG, the right hand was faster than the left in
the first (meansd, p value: 439.2551.89 vs. 488.5874.90, p
Conclusions:
We present evidence that 24-hour cast immobilization leads to a differentiation of the performances of the right and left hands in
very simple motor tasks. Its a novel finding that such a short period of immobilization disrupts the typical non-dominance for
these tasks.
Keywords: choice reaction time task, finger tapping, hand dominance, immobilism, motor control
Financial Support: The first author is supported by CNPq.
Resumo:27-063
ASTROGLIAL ALTERATIONS IN RATS SUBMITTED TO OKADAIC ACID-INDUCED DEMENTIA MODEL.
Bernardi, C. ; Costa, A. P. ; Tramontina, A. C. ; Biasibetti, R. ; Wartchow, K. M. ; Batassini, C. ; Tortorelli,

L. S. ; Gonalves, C. A.
UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL, UFRGS
Objectives:
Alzheimers disease (AD) is the most common form of dementia. The histopathological marks of the disease are the
formation of senile plaques, caused by the extracellular accumulation of amyloid fibrils in the brain, and also by the intraneuronal
aggregates of neurofibrillary tangles, which leads to progressive brain dysfunction, and astrocytosis. In vitro exposure to okadaic
acid (OA), a potent inhibitor phosphatase 1 and 2A, leads to the deposition of -amyloid, subsequent neuronal degeneration. In
this study, our aim was to evaluate spatial cognitive deficit and astroglial alterations in rats submitted to the okadaic acid-induced
dementia model, particularly with regard to astroglial protein markers, GFAP and S100B.
Thirty male Wistar rats (90 days old) were divided into 2 groups: sham (N = 14) and OA (N = 16). OA (100 ng) was
intrahippocampal infused, the rats received a single bilateral infusion of 1 L OA or vehicle (Hanks balanced salt
solution). Twelve days after surgery, rats were submitted to training and test in the Morris water maze. S100B and GFAP
contents measured by ELISA. The experiments statistically evaluated by Student-test, assuming p < 0.05. The escape latency
parameter in the water maze task was evaluated by repeated measures analysis of variance, assuming p < 0.05. The Morris water
maze was used to evaluate reference memory between two groups. There was a decline in the average time find to platform
(escape latency) from 4 onwards in the sham group. In addition, OA rats spent less time in the target quadrant, as compared to the
sham group. The time to find the platform was significantly lower in the OA group, as well as the number of crossings over the
platform location was less compared with sham group. A significant decrease in cerebrospinal fluid (CSF) S100B, a calcium-
binding protein predominantly expressed and secreted by astrocytes, was observed in the OA treated group. Assuming that
extracellular S100B has neurotrophic activity, this reduction could indicate impairment in astroglial function in some brain
regions in OA-treated rats. OA did not change S100B immunocontent in both brain regions, cortex and hippocampus. GFAP is a
specific astrocyte marker and its increase is taken as a sign of astrogliosis. In this study a significant increase in GFAP contents
was found in hippocampus in the OA group, this effect was not found in cortex.
Conclusions:
In conclusion, we have provide evidence that intrahippocampal-injected OA rats confirm the spatial cognitive deficit and
astroglial alterations, particularly GFAP increase in the hippocampus and the decrease in CSF S100B. Findings contribute to
understanding diseases accompanied by cognitive deficit and the OA model of dementia.
Keywords: DEMENTIA, OKADAIC ACID, ASTROCYTES, S100B, GFAP
Financial Support: CNPq, INCT for Excitotoxicity and Neuroprotection
Resumo:27-064
DEVELOPMENTAL OUTCOMES OF FLAXSEED INTAKE IN A RAT MODEL OF PERINATAL CEREBRAL
HYPOXIA-ISCHEMIA
Mucci, D. B. ; Souza, A. S. ; Rebello, N. S. ; Rodriguez, C. B.

Depto de Nutrio e Diettica/Instituto de Nutrio, UFRJ
Objectives:
The low oxygen supply (hypoxia) during the neonatal phase is the most frequent cause of permanent brain damage. The
polyunsaturated fatty acids (PUFA) n-3 are essential for neural development and exert beneficial effects on several injury
cascades involved in perinatal brain injury. The flaxseed (Linum usitatissimum) is one of the best AGPI n-3 vegetable sources.
The aim of this study is to access the effects of maternal flaxseed intake on the growth, aversive and spatial memory of the
offspring submitted to hypoxia-ischemia (HI).
Wistar rats were divided in 2 groups fed isocaloric and normolipidic diets containing different fat compositions: Control Group
(C), with soy oil as its lipid source; Flaxseed Group (F), with higher levels of PUFA n-3 and flaxseed as the lipid source. The
experimental diets were fed to mothers during pregnancy and lactation and a standard diet was given to the post-weaning pups for
4 weeks. The male pups were separated in 3 subgroups within each dietary group: HI (CHI and FHI), in which pups underwent
right carotid ligation, followed by 8% O2 for 90 minutes on postnatal day (P) 7; Sham (CSh and FSh), the false surgery group;
Control (CC and FC). The pups weight and food intake were evaluated from P21 (weaning) to P49. Between P30 and P40, the
animals were submitted to the behavioral tests Morris water maze (MWM) and passive avoidance test (PAT). Data are presented
as mean SD. Data were analyzed using t-test or Anova one way followed by Bonferroni post-hoc test and p0.05 value was
considered to be significant. Both HI groups presented inferior growth compared to the other groups from P21 to P49; FHI
(n=14) had even lower growth results than CHI (n=16) (P21: CHI: 41,516,96g, FHI: 33,225,84g; P28: CHI:61,4311,56g,
FHI: 46,8912,60g; P35: CHI: 101,7416,86g, FHI: 73,9115,92g; P42: CHI: 142,5822,43g, FHI:149,9123,45g; P49: CHI:
183,1226,98g, FHI: 149,9123,45g, p
Conclusions:
The results suggest that HI causes growth deficit, especially in animals fed the flaxseed diet, even though FHIs food intake was
significantly higher. Moreover, HI impaired spatial memory, but the flaxseed was able to prevent damage concerning aversive
memory.
Keywords: flaxseed, hypoxia-ischemia, neonatal
Financial Support: Sources of research support: CAPES, CNPq
Resumo:27-065
HUMAN BRAIN VOLUMETRY: RELATIONSHIP BETWEEN GLOBAL AND CORTICAL VOLUMES BY MRI IN
POST MORTEM
Emdio, R. S. 1,2,3; Santos, G. A. B. D. 2,3; Vieira, G. 4,1; Neves, R. D. C. 2; Carreira, L. L. 2; Grinberg, L. T.

2,3,5
; Amaro Junior, E. 1
1
Departamento de Radiologia HC FMUSP, INRAD - HC FMUSP
2
Fisiopatologia no Envelhecimento GEROLAB FMUSP, GEROLAB FMUSP
3
Instituto Israelita de Ensino e Pesquisa Albert Einstein , IIEPAE
4
Programa Interunidades de Ps-Graduao em Bioinformtica, PIPGBI
5
University of California, San Francisco Memory and Aging, UCSF
Objectives:
Ageing is accompanied by morphological and functional changes due to progressive adaptation of progression many metabolic
processes. A great effort has been done to understand the relationship between aging and neurodegenerative processes. Imaging
examinations are non-invasive and are considered promising tools for addressing this problem. However, the interpretation of
imaging findings is mostly based on assumptions since many limitations prevent reliable imaging-histopathological correlation
studies. A group of multidisciplinary researchers combined the resources of the Human Brain Bank of Aging Brain Study Group
from University of Sao Paulo Medical School and the Institute of Radiology from the same University to create a platform to
overcome these limitations. The idea is to use in situ postmortem MR images to avoid movement artifacts and provide a close
measure to histological analysis. The present study aims to identify specific changes in measures of cortical volume from
segmented gray matter in elderly subjects without cognitive impairment.
We analyzed in situ post-mortem images of five healthy subjects (3 males, mean age 64 7.5 y). The clinic functional status was
assessed through a standardized semi-structured interview, using the clinical dementia rating, and all had score 0, they are all
individuals without dementia. All the subjects were submitted to a postmortem and in-situ 3T MRI examination. We use this
image parameters: T1-TFFE: TR = 6,3 ms, TE = 2,9 ms; 1,0 mm x 1,0 mm x 1,0 mm; 1 NEX; FOV 24 x 240 cm. The cortical
volumes were measured using an automatic algorithm for tissue segmentation and quantification - Freesurfer (Dale et al., 1999;
Fischl et al., 1999; Fischl and Dale, 2000). The cortical analysis was based on segmentation of different brains regions, extraction
of the skull and other parts of intracranial space and automatic segmentation of cortical and subcortical regions. This method uses
both the signal strength of the MRI T1-weighted, and the continuity of the surfaces to produce representations of cortical
thickness. We estimated the percent change of volume (variability) between all subjects in relation to overall brain volume. The
analysis was performed using only individuals without cognitive impairment in order to avoid bias in the analysis due to tissue
loss related to neuronal disorders reported during life.The values of percentage of cortical volumes measured in relation to
cortical volume were as follows: subject 1: 38.85%; 2: 42.411%; 3: 36.329; 4: 42.923%, 5: 44.715%. The numerical data were
obtained using nonparametric tests showed a p value
Conclusions:
This result suggests that there is a significant variation of cortical volume related to the overall volume of the brain. Studies based
on measures of cortical volume to detect neuronal abnormalities should consider this variation when performing analysis of
statistical power. Moreover, this result show that a small change in percentage of brain cortex is compatible with preserved
cognitive abilities during the aging process.
Keywords: MRI, Brain, Volume
Financial Support: IIEP Albert Einstein
Resumo:27-066
MATERNAL SEPARATION CONTRIBUTES TO A FASTER LEARNING IN ADULTHOOD
Iyomasa, M. M. 1; Limonte, F. H. 1,3; Fachim, H. A. 2,3; Pereira, M. T. R. 1; Rosa, M. L. N. M. 1,3

1
Faculdade de Medicina de Catanduva, FIPA
2
Fac. Filosofia Cincias e Letras de Ribeiro Preto - USP, FFCLRP-USP
3
Instituto de Neurocincia e Comportamento, INeC
Objectives:
Maternal separation (MS) has been proposed as an animal model of early life stress and the subsequent development of
depression. The aim of this study was to investigate the long-lasting behavioral effects induced by prolonged maternal separation
in rats.
Male Wistar rats have been used. In the MS the pups (n=8) underwent a daily-3h separation from their mothers from the first
postnatal day to weaning (PND1-21) and the control pups (non-separated NS, n=8) were left undisturbed. Following weaning
the animals were housed in groups of 4/cage for 5 weeks before testing: Spatial memory was evaluated on Morris Water Maze
and Recognition memory was evaluated by the novel object discrimination test. Spatial memory: A circular pool (diameter = 104
cm; height = 50 cm) containing water at 22 2C and a platform (diameter = 9 cm) submerged 1 cm below the surface at the
midpoint of one quadrant was used. The animals were submitted to six training sessions (90 seconds) per day for 4 days and the
latencies of escape to find the platform recorded. At the end the platform was removed and the time spent swimming on that
quadrant was recorded (probe trial). Recognition memory: on the first day rats were submitted to a habituation session on the
training arena for 5-min. On the following day, rats were given 5-min training trial in which they were exposed to two identical
objects (A1 and A2). On the short-term memory (STM) testing trial (90 min after training), rats were allowed to explore two
objects: a familiar one (A) and a different one (B) for 5-min. On the long-term memory (LTM) testing trial (24 hours after
training), rats were allowed to explore two objects: (A) and a third different one (C) for 5-min. A recognition index was
expressed by the ratio TB/(TA+TB) or TC/(TA+TC). In both tests MS and NS groups were compared by Student t-test (p
Conclusions:
Pups separated from their mothers for long periods (3 hours/day until weaning) do not display spatial or recognition memory lost
in adulthood. Contrasting to this, MS may induce a faster learning as the animals spent a very short time to find the platform on
the Morris water-maze. In addition, MS may induce a more active behavior in adulthood as the animals spent a higher time
exploring both objects on the recognition memory test.
Keywords: Maternal Separation, Morris Water Maze, novel object discrimination test, Rats, learning and memory
Financial Support: FAPESP and Padre Albino Foundation
Resumo:27-067
IMPAIRED COGNITION AND BDNF LEVELS IN CHILDREN VICTIMS OF TRAUMA COMPARED WITH
CONTROLS
Wollenhaupt-aguiar, B1,2,3; Bcker, J. 2,4,3; Ceresr, K. 2,3,4; Kapczinski, N. 2,3,4; Pfaffenseller, B. 5,2,3;
Panizzutti, B. S. 2,4,3; Kapczinski, F. 2,3,4; Kauer-sant'anna, M. 2,3,4
1
Programa de Ps Graduao em Cincias Mdicas/UFRGS, PPGCM/UFRGS
2
Instituto Nacional de Cincia e Tecnologia Translacional, INCT-TM
3
Laboratrio de Psiquiatria Molecular/HCPA, HCPA
4
5
Programa de Ps Graduao em Cincias Biolgicas:Bioqumica, PPGBIOQ/UFRGS
Objectives:
Development of psychiatric disorders and cognitive impairment in adulthood is associated with the exposure to traumatic events
during childhood. However, few studies have examined cognitive function in school-age children with a history of maltreatment,
abuse or neglect. Also, the brain derived neurotrophic factor (BDNF) is the most studied member of neurotrophic factors and is
involved in neuronal plasticity and neuroprotection, as well as in the pathophisiology of psychiatric disorders such as in bipolar
disorder and schizophrenia. Our study aim to examine cognitive function, psychiatric diagnosis and serum BDNF levels in
children victims of trauma compared to controls matched for age and gender.
We recruited 30 children with childhood trauma and 30 matched controls. The evaluation included a socio-demographic protocol,
cognitive tests and a diagnostic interview of psychiatric disorders according to DSM-IV. Serum BDNF levels were assessed with
sandwich ELISA Kit.Differences among patients and controls in all experiments were determined by independent sample t test.
Results: Analysis showed a high prevalence of psychiatric symptoms in those with childhood trauma (p
Conclusions:
There is a high prevalence of psychiatric disorders in children victims of trauma and this seems to be associated with worse
cognitive performance and higher levels of BDNF in this sample.
Keywords: BDNF, COGNITION, CHILDREN, TRAUMA
Financial Support: INCT-Translacional em Medicina, FIPE-HCPA, CNPQ
Resumo:27-068
BEHAVIOUR IN THE OPEN FIELD AND ELEVATED PLUS MAZE, IN WISTAR RATS AT 60 DAYS OF AGE,
WITH CONTROLLED INGESTION OF ERVA MATE TEA (ILEX PARAGUARIENSIS).
Padoin, M. J. ; Fronza, J. L. ; Cantelli, K. R. ; Silva, H. M. V.

Centro de Cincias Biolgicas e da Sade, UNIOESTE
Objectives:
Ilex Paraguariensis is a plant used in the form of chimarro, tea and terer. Few studies have addressed the influence of this
tea on behaviour. The effect of controlled ingestion of erva-mate tea on the behaviour of 60-day old Wistar rats was investigated
in the open field and the elevated plus maze.
Control Group (CG)(n=12 males,2 females) received water; Ingestion Group (IG) (n=12 males,12 females) received erva mate
tea. Erva mate was obtained from a company in the city of Cascavel, PR. The tea was prepared daily by adding the erva mate (g)
to boiling distilled water (mL) at a ratio of 20:1 (20 water to 1 of herb); this was filtered, cooled and offered to the IG. The
animals were given the tea for 2 hours daily and intake per rat was evaluated, preceded by a period of 4 hours without
liquids.From the age of 60 days the rats received the tea for 60 days, at which point they were assessed in the Open Field test, in
which the behaviours analysed were: locomotion in the periphery and centre, remaining stationary,rearing and grooming; and in
the Elevated Plus Maze, where the behaviours examined were: time spent on the open arm, time spent on the closed arm, head
dipping on the open arm and head dipping on the closed arm. Latency, frequency and duration were determined. For male rats in
the Open Field, the latency to grooming in CG (42.35.4) was lower than in IG (70.410.0), although the frequency for the
IG (20.73.5) was greater than that of CG (5.61.0). The frequency of locomotion in the periphery for CG (9.80.9) was
higher than that of IG (5.90.6), likewise locomotion in the centre (CG=7.11.0; IG=3.70.6), and remaining stationary
(CG=4.30.6; IG=2.80.5). In the Elevated Plus Maze test, the males of CG (15.82.3) presented a greater frequency of
head dipping on the open arm than IG (9.32.2), besides spending more time on the open arm (GC=130.117.5;
GI=61.314.0), and spent more time engaged in head dipping on this arm (CG=49.18.4; IG=22.76.8). Consequently, the
IG males (235.214.4) spent more time on the closed arm than CG males (165.017.6), as well as presenting more head
dipping on the closed arm (IG=66.96.1; CG=48.36.8). Among females in the Open Field, the latency to move to the centre
was shorter in IG (1.00.0) than in CG (42.416.4). The CG females (15.11.6) exhibited a greater frequency of
locomotion in the periphery then IG females (9.00.9), as well as stopping more frequently (CG=12.61.0; IG=5.11.0).
With regard to the duration of behaviours, it was noted that rearing occurred more among CG females (29.44.6) than IG
females (17.02.9). In the Elevated Plus Maze test, IG females (5.71.7) entered the closed arm earlier than CG females
(17.46.1). Concerning the frequency of behaviours, CG females (10.31.7) demonstrated more head dipping on the open
arm than IG females (5.81.5), while in terms of duration, CG females (79.814.1) spent more time on the open arms than
those of the IG (39.911.0), and spent more time engaged in head dipping on the open arm (CG=27.17.5; IG=8.41.9).
The IG females (256.211.4) spent more time on the closed arm than CG females (213.114.5). ANOVA was performed
followed by Newman-Keuls test when appropriate, with p<0,05.
Conclusions:
The results show that in both male and female rats, at 60 days of age, the controlled ingestion of erva mate tea lead to an increase
in fear and anxiety in the behavioural tests used.
Keywords: Ilex paraguariensis, open field, elevated plus maze
Financial Support: UNIOESTE
Resumo:28-011
SURVIVAL AND FILAMENTATION OF ESCHERICHIA COLI CELLS AND ELECTROPHORETIC PROFILE OF

PLASMIDS EXPOSED TO LOW-INTENSITY INFRARED LASER
Canuto, K. S. 1; Sergio, L. P. S. 1; Marciano, R. S. 1; Martins, M. 1; Guimares, O. R. 1; Polignano, G. A. C.

1
; Geller, M. 1; Paoli, S. 1; Paoli, F. 3,1; Fonseca, A. S. 2,1,4
1
Centro de Cincias da Sade, UniFESO
2
Departamento de Cincias Fisiolgicas/Instituto Biomdico, UNIRIO
3
Departamento de Morfologia/Instituto de Cincias Biolgicas, UFJF
4
Departamento de Biofsica e Biometria/Instituto de Biologia, UERJ
Objectives:
Low-intensity infrared lasers are used for treating of different oral lesions based on its phototermal, photochemical and
biostimulant effects on biological tissues. However, data about possible adverse effects of this laser on DNA are scarce yet. The
objective of this work was to evaluate effects of low-intensity infrared laser on survival and filamentation of Escherichia coli
cells and on electrophoretic profile of bacterial plasmids.
E. coli AB1157 (wild type), BH20 (fpg mutant) and BW9091 (xthA mutant), in exponential and stationary growth phase, and
pBSK plasmids were exposed to infrared laser (808 nm) following clinical protocol for treating of pain (60 and 120 J/cm2).
Bacterial suspensions and plasmids did not expose to laser were used as controls. Aliquots of bacterial cultures were spread onto
Petri dishes containing nutritive medium, incubated (37 oC, 18 hours) and determined the survival fractions (SF). Other aliquots
were spread onto microscopic slides, stained by Gram method and observed under light microscopy (40 x). Analysis of images
was carried out by Image Proplus software (two slides for fluency, 3 fields for slide, 200 cells for field). After, percentages of
bacterial filaments (%BF) were determined. Agarose gel electrophoresis (0.8%; 7V/cm) was carried out and the plasmidial forms
were visualized on transiluminator and the images were captured for quantification of percentage of supercoiling form (%SC).
Data obtained for SF in exponential and stationary phase, in the higher fluency, for AB1157, were (XSD): 1.10.21 and
1.80.28, for BH20: 1.00.12 and 1.00.24, for BW9091: 1.10.26 and 1.00.25. Data for %BF, in exponential phase, were for
AB1157: 2.30.58 (control) and 7.30.58 (120 J/cm2), for BW9091: 19.01.00 (control) and 33.72.08 (120 J/cm2), for BH20:
1.00.28 (control) and 5.71.53 (120 J/cm2); in stationary phase were, for AB1157: 2.70,58 (control) and 3.30.58 (120 J/cm2),
for BW9091: 67.71.53 (control) and 64.74.16 (120 J/cm2), for BH20: 1.30.48 (control) and 4.30.38 (120 J/cm2). For %SC:
88.70.38 (control) and 83.14.09 (120 J/cm2). Infrared laser exposure significantly (pE. coli cultures in exponential growth
phase.
Conclusions:
Results indicate that exposition to infrared laser 808 nm in clinical protocol used for treating oral pain could not alter the
electrophoretic profile of plasmids but alter survival of wild type bacterial cultures in stationary growth phase and induce
bacterial filamentation in E. coli cultures in exponential growth phase.
Keywords: Escherichia coli, DNA, Filamentation, Electrophoresis, Laser
Financial Support: FAPERJ, UniFESO, UNIRIO
Resumo:28-012
EFFECTIVENESS OF LASER PHOTOBIOMODULATION (660NM) AND POLARIZED LIGHT (400-2000NM) ON
CUTANEOUS WOUNDS HEALING IN HYPOTHYROID RATS.
Santos, G. B. D. S. 2; Weyll, B. M. . P. 2; Paraguass, G. M. 2; Rodriguez, T. T. 1; Ramalho, M. J. P. 1;

Pinheiro, A. L. B. 2; Ramalho, L. M. P. 2
1
Depto de Biorregulao - Fisiologia, ICS - UFBA
2
Faculdade de Odontologia, FOUFBA
Objectives:
The aim of this study was to assess influence of laser photobiomodulation (660nm) and polarized light (400-2000nm) on
cutaneous wounds healing in hypothyroid rats at dosages of 20 or 40J/cm.
Forty adult male Wistar rats, weighing 230-260g, were randomly distributed into two groups (Euthyroid and Hypothyroid), of
twenty-two animals each. Each group was then divided into five subgroups based on treatment applied: Untreated Control; Laser
20 J/cm and 40 J/cm; and Polarized Light source 20J/cm and 40J/cm. Hypothyroidism was induced in rats with
propylthiouracil (0,05g/100mL) administered orally for 4 weeks and maintained until the end of the experiment. Standard
cutaneous wound (1 by 1 cm) was created on the dorsum of each animal under general anesthesia. Immediately after surgery, the
wounds were either irradiated transcutaneously with Laser (Kondortech, Brazil, 660nm) applied to four different points around
the wound every 48 hours throughout seven days or illuminated with a Polarized Light source (400-2000nm; Bioptron,
Switzerland) kept at 10 cm focal distance in order to standardize photobiomodulation. The rats were killed on the eighth day.
Specimens were removed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE) or Sirius red-stained
sections by light microscopy. Statistical analysis was performed using the Fisher's exact test non-parametric method. The
cutaneous wounds of hypothyroid rats showed delayed healing process characterized by reduced thickness of epithelial layers,
incipient formation of disorganized collagen fibers (FISHER, p=0.0276), when compared to the euthyroid group. The use of both
the Laser and Polarized Light on hypothyroid rats increased the amount of fibroblasts and the thickness of collagen fibers,
especially on the Laser 20J/cm group. Euthyroid rats have still demonstrated more regular collagen fibers pattern than
hypothyroid rats.
Conclusions:
It was therefore concluded that hypothyroidism delays wound healing and both Laser Photobiomodulation and Polarized Light at
dosages 20J/cm had improved the healing process in hypothyroid rats.
Keywords: laser, hypothyrodism, LED
Financial Support: Cnpq, Fapesb e PIBIC UFBA.
Resumo:28-013
IMMUNOHISTOCHEMICAL ANALYSIS OF CATALASE AND VEGF EXPRESSION IN THE VISUAL SYSTEM OF
THE CRAB UCIDES CORDATUS FOLLOWING EXPOSURE TO UV-B RADIATION
Fusco, M. A. 1; Wajsenzon, I. 1; Hollman, G. 1; Wyatt, N. 1; Miguel, N. C. D. O. 2; Leito, A. C. 3; Allodi, S.

1
3
Laboratrio de Radiobiologia Molecular, IBCCF

Instituto de Biofsica Carlos Chagas Filho, IBCCF, CCS, UFRJ
2
5Programa de Biologia Celular e do Desenvolvimento, ICB, CCS, UFRJ
1
Objectives:
The generation of free radicals caused by the exposure to ultraviolet B (UV-B) radiation is a well known phenomenon. The role
of catalase in the mechanism of antioxidant defense to the visual system is a good predictive of the extent of the light-induced
injury to the tissue. In addition, vascular endothelial growth factor (VEGF) has been implicated as a neuroprotector and
neurotrophic factor, besides its well known role in the mitosis of endothelial cells. Using the optic lobe of the crab as a model for
the study of the visual system, the objective of this study was to evaluate the expression of VEGF following exposure to UV-B
radiation, supported by the expression of catalase.
Five experimental male animals of the species Ucides cordatus were exposed to UV-B (Vilber Lourmat UV-B lam 312 nm) for
30 minutes per day for five days (total dose 4 W/m2). Five animals, used as controls, were not exposed to UV-B radiation,
although they were kept for the same 30 minutes under visible light (Phillips TLT 40 W/75). Two days after the last time of
exposure, animals from both irradiated and control groups were anesthetized by chilling and their optic lobes were dissected and
fixed in 4% paraformaldehyde overnight followed by cryoprotection in 5, 10 and 20% sucrose, successively, overnight. The optic
lobes were embedded in optimal cutting temperature compound and sections at a thickness of 5 m longitudinal to the long axis
of the stalk were obtained and mounted on poly-L-lysine-coated slides. For the immunohistochemical reactions the following
primary antibodies were used: rabbit anti-VEGF (Millipore) and rabbit anti-catalase (Chemicon) at 1:100 and 1:50 dilutions,
respectively. The secondary antibody used was anti-rabbit Alexa 546 (Invitrogen) at 1:600 dilution. Slides containing the retina,
lamina ganglionaris, external and internal medulla cells were evaluated under the confocal microscope and images were recorded
using the software Cell . The pattern of cytoplasm staining for VEGF and catalase in both the control and irradiated optic lobes
were quantified using the software Image Pro Plus. The statistical analysis comparing the results from the histomorphometry of
the treated and the control optic lobes was calculated using the software Graphpad Prism 5.0 (unpaired t test; p value
Conclusions:
UV-B induced-injury, related to the elevated labeling profile for catalase, is also accomplished by the inhibited expression of
VEGF in the optic lobes of the crab Ucides cordatus.
Keywords: radiation, UVB, Ucides cordatus, caspase, VEGF
Financial Support: CAPES, FAPERJ, CNPq
Resumo:28-014
CLINICAL EVALUATION OF THE EFFECTS OF LASER AND LED PHOTOBIOMODULATION ON CUTANEOUS
WOUND HEALING IN HYPOTHYROID RATS
Santos, G. B. D. S. 2; Paraguass, G. M. 2; Cerqueira, N. S. 2; Guarda, M. G. 2; Jesus, V. C. 2; Rodriguez, T.

T. 1; Ramalho, M. J. P. 1; Pinheiro, A. L. B. 2; Ramalho, L. M. P. 2
1
Depto. de Biorregulao - Fisiologia, ICS - UFBA
2
Faculdade de Odontologia - Universidade Federal da Bahia, FOUFBA
Objectives:
The aim of this study was to assess clinically the influence of low-level laser therapy (LLLT) and Light-Emitting Diodes (LED)
on healing process on dorsal cutaneous wounds of hypothyroid rats at dosages of 24J/cm.
Seventy-two male Wistar rats, weighing 200-230g, were randomly distributed into two groups (Euthyroid and Hypothyroid), of
thirty-six animals each, and these were divided into six groups, according to the type of phototherapy received (Untreated
Control; Laser 24 J/cm and LED 24J/cm) and time of death (7 or 14 days). Hypothyroidism was induced in rats with
propylthiouracil (0,05g/100mL) administered orally for 4 weeks and maintained until the end of the experiment. Standard
cutaneous wound (1 by 1 cm) was created on the dorsum of each animal under general anesthesia, and was left without suturing
or dressing. Immediately after surgery, the wounds were either irradiated transcutaneously with Laser (24J/cm, 660nm,
GaAlAs, 40mW, CW, f=4mm, Twin Flex Evolution, MMoptics, Brazil), applied to four different points around it, or irradiated
over the wound area in a single point with LED (24J/cm, InGaAlP, 630nm 150mW, CW, f=0,5 cm, FisioLED, MMoptics,
Brazil), and was repeated every other day for 7 or 14 days. Distances between the edges of the wound in its largest dimensions
were obtained using a digital caliper both on the day of the surgical procedure, as of the death of animals to assess cicatricial
contraction. Statistical analysis was performed using the Mann-Whitney non-parametric method, considering a statistical
significance level of 5% (p 0.05). Hypothyroid rats that did not undergone phototherapy, had wound cicatricial contraction to a
lesser extent than euthyroid rats only in the period of 14 days (Mann-Whitney, p=0.025). There was significant difference
between euthyroid and hypothyroid rats undergone Laser and LED phototherapy, especially in 7 days, regarding to macroscopic
analysis of wound contraction (Mann-Whitney, p=0,037 and p=0,025).
Conclusions:
This study has shown that hypothyroidism delays wound healing and Laser and LED photobiomodulation at 24 J/cm improves
the cutaneous wound healing in hypothyroid rats.
Keywords: LASER, LED, HYPOTHYROIDISM
Financial Support: CNPq, FAPESB, PIBIC/UFBA
Resumo:28-015
VALIDATION OF A METHOD TO DETERMINE THE STANNOUS ION IN RADIOPHARMACEUTICAL KITS
Dadda, A. S. ; Teixeira, A. C. ; Leite, C. E. ; Campos, M. M. ; Jeckel, C. M. M.

INSTITUTO DE TOXICOLOGIA, PUCRS
Objectives:
The stannous ion (Sn+2) is mainly used in the form of stannous salt, in order to reduce the pertechnetate ion (TcO4-), during the
production process of the radiopharmaceutical 99mTc. The easy oxidation of Sn+2 by the action of different agents may result in
loss of labeling efficiency, thus requiring the determination of its concentration in the final formulation. Voltammetry represents
one of the most appropriate methods for the selective determination of Sn+2, in the presence of its oxidized form (Sn +4). The aim
of this study was to develop and validate an analytical and selective method for quantification of Sn +2.
We have employed a 757 VA Computrace Polarography (Metrohm), composed of three types of electrodes, which are: a
multimode mercury electrode (MME), a platinum auxiliary electrode, and a reference electrode silver/silver chloride (Ag/AgCl).
The sample used for analysis was the lyophilized radiopharmaceutical MIBI (2-methoxy-2-isobutyl isonitrile). The analytical
procedure proved to be sensitive, selective, precise, accurate and linear (regression coefficients greater than 0.999) in the range of
10 to 35.0 g/mL for Sn+2. The limit of detection was 3.0 g/mL, and the limit of quantification was 4.41 0.49 g/mL. The
selective detection of stannous ion was further recorded using the same sample during three weeks. The gradual drop in the Sn +2
current was assessed at 1, 2, 13 and 23 days after the preparation of test solutions, with decrease percentages of 12 %, 28 %, 39
%, and 100 %, respectively. The current decreases due to the formation of the Sn +4 (oxidized form of Sn+2). The formation of this
element does not interfere in the analysis because it has a different potential.
Conclusions:
The method presented high sensitivity and might be considered a reliable parameter for the selective determination of Sn +2 in
radiopharmaceutical MIBI solutions, without interference of degradation products of Sn +2.
Keywords: Voltametry, radiopharmaceutical products, liofilized kits, stanous ion, eletrochemistry
Financial Support: CNPq e PUCRS
Resumo:28-016
SPEM (SINGLE PHOTON EMISSION MICROSCOPY): A NEW TECHNOLOGY OF MULTI-PINHOLE SPECT
(SINGLE PHOTON EMISSION COMPUTED TOMOGRAPHY) IMAGING FOR SMALL ANIMALS.
Reis, M. A. 1,2; Mejia, J. 1; Batista, I. R. 1,2; Shih, M. C. 1,3; Fu, G. 3; Cabral, F. R. 1; Barboza, M. R. F. F. D.
1
; Nogueira, S. A. 1; Amaro-jr, E. 1; Chen, C. T. 3; Abilio, V. C. 2; Bressan, R. A. 1,2
1
Instituto Israelita de Ensino e Pesquisa Albert Einstein, IIEPAE
2
Laboratrio Interdisciplinar de Neurocincias Clnicas, LiNC - UNIFESP
3
University of Chicago, UC
Objectives:
Currently, techniques of molecular imaging such as SPECT (Single Photon Emission Computed Tomography) have been used in
Nuclear Medicine for diagnoses and clinical research. However, for pre-clinical research - in which small animals are mostly
used (usually mice and rats) the spatial resolution and sensitivity achieved is low due to the reduced size of these animals. In this
sense, an alternative to increase the sensitivity and spatial resolution of this technique is to use a multi-pinhole collimation
system. In this study, we present the Single Photon Emission Microscope (SPEM), a state-of-art instrument for small animal
SPECT imaging, based on multi-pinhole collimation and high sensitivity Electron-Multiplying CCDs, in combination with
appropriate Maximum Likelihood-based software tools, to obtain 3D images of small animal organs.
Methods: The SPEM (developed by the Department of Radiology and Medical Physics Committee from University of Chicago
and installed at the Pre-Clinical Image Center of CETEC IIEPAE) consists of two separate imaging devices based on high
spatial resolution columnar scintillators, in combination with high sensitivity, high resolution Electron Multiplying CCD
(EMCCD) cameras. The equipment uses imaging devices with 7 and 19 pinhole collimators, respectively, each pinhole with 200
m of diameter. During the image acquisition procedure, the mouse is placed in an animal holder and is maintained in the vertical
position to minimize the oscillation of the internal organs. This holder rotates in front of the imaging detector. After acquisition of
the planar projections, a specially designed software tool based on the Maximum Likelihood algorithm iteratively searches for the
model of the radiopharmaceutical distribution on the target which better explains the recorded images. The radiopharmaceuticals
used in this protocol were radiotracers for kidneys, heart and thyroid: 99mTc-DMSA, 99mTc-MIBI and 99mTc-Pertecnetate,
respectively. These agents were injected into the tail veins at doses of 2 mCi/0,3 mL 30 minutes before the image acquisition.
During this injection procedure and data acquisition, the animals were maintained under ketamine and xilazine anesthesia.
Results: Images of kidneys, heart and thyroid obtained with SPEM will be presented.
Conclusions:
This study shows the performance of the Single Photon Emission Microscope (SPEM). This new technology of SPECT will
allow the development of a new molecular image facility in Brazil providing high resolution images. Experimental protocols for
acquisition of brain images using dopaminergic and serotoninergic radiopharmaceuticals are currently under development by our
group. Undoubtedly, this new tool, associated with the use of different radiotracers, can advance our knowledge about a variety
of diseases, their pharmacological treatments and the development of potential new therapeutical agents.
Keywords: SPEM, Small-Animal SPECT, multi-pinhole SPECT
Financial Support: FAPESP; ABADHS (Ass. Beneficente Alzira Denise Hertzog da Silva)
Resumo:28-017
EFFECTS OF LOW POWER LASER (GAAS) ON PARAMETERS OF OXIDATIVE STRESS IN AN ANIMAL
MODEL OF MUSCLE TRAUMA.
Silveira, P. C. L. ; Matos, F. ; Scheffer, D. D. L. ; Farina, M. ; Latini, A.

Objectives:
Recent studies demonstrate that low-level laser therapy (LLLT) modulates many biochemical processes, especially the decrease
of oxidative stress accelerating the muscular healing process. The aim of this study was to investigate the effect of LLLT on
oxidative stress parameters in an animal model of muscle trauma.
Methods: Male Wistar rats were randomly divided into the following four groups (n = 6): controls (uninjured muscle), muscle
injury without treatment animals, untreated animals with LLLT (AsGa) 3 J/cm2, and muscle injury animals with LLLT (AsGa) 3
J/cm2. Gastrocnemius injury was induced by a single blunt-impact trauma. LLLT was used 2, 12, 24, 48, 72, 96, and 120 hours
after muscle-trauma. The parameters namely, antioxidant enzymes activities, including superoxide dismutase (SOD), glutathione
peroxidase (GPX) and catalase (CAT), and lipid (TBARS) and protein oxidation (carbonyl groups) measurements were assessed.
Results: The results showed marked lipid and protein oxidation in muscle-injured animals (0.08 0.006; 0:34 0.02 nmol/mg
protein, respectively) compared to controls (0.05 0.003; 0:21 0.01 nmol/mg protein, respectively) and to muscle-injured
animals with LLLT 3 J/cm2 (0.05 0.003; 0:23 0.01 nmol/mg protein, respectively). The same profile was observed in injured
rats for the antioxidant enzymes activities, which were significantly increased (SOD: 0.18 0.02; CAT: 3:55 0:44; GPx: 3:00
0.31 U/mg protein), when comparing to controls (0.09 0.01, 2.19 0.19; 02.07 0.20 U/mg protein) and to muscle injury with
LLLT 3 J/cm2 group (0.08 0.01; 1:39 0.25, 1.72 0.20 U/mg protein).
Conclusions:
Our results showed that LLLT is effective in the reduction of the oxidative stress after 5 days of mechanical trauma.
Keywords: LOW POWER LASER, MUSCLE TRAUMA, OXIDATIVE STRESS
Financial Support: UFSC, CAPES
Resumo:28-018
COMBINED USE OF LOW LEVEL LASER THERAPY AND CYCLOOXYGENASE-2 SELECTIVE INHIBITION ON
SKIN INCISIONAL WOUND REEPITHELIALIZATION IN MICE: A PRECLINICAL STUDY
Felix, M. B. M. 1; Santuzzi, C. H. 2; Calixto, K. V. 1; Gouvea, S. A. 2; Nogueira, B. V. 2; Rodrigues, A. N. 1;

Goncalves, W. L. S. 1,2
1
Centro Universitario do Espirito Santo, UNESC
2
Universidade Federal do Espirito Santo, UFES
Objectives:
Low level lasertherapy (LLLT) and cyclooxygenase-2 (ICOX-2) selective inhibitors have been widely used to modulate
inflammatory response; however, their effects on wound reepithelialization are not well understood. In this study the objective
was to evaluate the isolated and combined effects of low level laser therapy and ICOX-2 in the reepithelization of skin incisional
wounds in mice.
We induced a 1-cm wound on the back of each mouse, which were divided into four groups (N = 20): control, lasertherapy,
treated with ICOX-2 and combined therapy. The animals in the celecoxib and combined therapy groups were treated with
celecoxib for 10 days before skin incision. The experimental wounds were irradiated with LLLT (He-Ne 632nm, dose: 4J/cm2) in
scanning for 12 seconds during three consecutive days in the lasertherapy and combined therapy groups. The animals were
sacrificed 3 days after surgery. Samples of the wounds were collected and stained (Masson's Trichrome) for histomorphometric
analysis. Both the LLLT group and the celecoxib group showed an increase in skin reepithelialization compared to the control
group; however, the combined therapy group showed no differences. As for keratinization, the LLLT and combined therapy
groups showed a reduction in keratinocytes compared with the control group.
Conclusions:
The results show that the use of LLLT He-Ne and ICOX-2 in isolation increases epithelial cells, but only LLLT reduced skin
keratinocytes. The combined treatment restores innate epithelialization and decreases keratinization in spite of accelerating
wound contraction with improvement in the organization of the wound in the skin of mice.
Keywords: COX-2 inhibitors, Epithelial cells, Keratinocytes, Lasertherapy, Wound healing
Financial Support: CDV-FACITEC
Resumo:28-019
EFFECT OF MATERNAL ETHANOL INTAKE DURING THE PREGNANCY AND LACTATION ON THE FEAR,
ANXIETY AND SPATIAL MEMORY IN PUPS AT DIFFERNT STAGES OF BRAIN DEVELOPMENT
Fiorillo, H. M. 1; lvares, R. M. 1; Graa, M. S. 1; Bortolani, V. 1; Iyomasa, M. M. 1; Rosa, M. L. N. M. 1,2

1
Faculdade de Medicina de Catanduva, FIPA
2
Institute of Neuroscience and Behavior USP Ribeirao Preto, INeC
Objectives:
The aim of this work was to investigate the effect of maternal ethanol intake during pregnancy and lactation on the fear and
anxiety behaviors and on spatial memory in pups at different stages of brain development.
Female Wistar rats (150g, n=16) were exposed to water (control group, n=8) or 5%-10% of ethanol (n=8), increasing 5% per
week (habituation), and 10% maintained for 60 days (chronic ingestion), corresponding to the period before mating (8 days),
mating (10 days), pregnancy (21 days) and lactation (21 days). After weaning the pups (n=61) were left undisturbed in their cages
(4/cage) until PND30 (control,n=8 and alcoholic,n=8), PND60 (control,n=8 and alcoholic,n=16) or PND90 (control,n=8 and
alcoholic,n=13) with food and water ad libitum. The levels of anxiety and the spatial memory were analyzed on the elevated plusmaze (EPM). Behavioral responses were scored for 5min: number of entries and time on the open arms, number of entries and
time on the closed arms, number of stretched attend postures (SAPs). Animals were retested after 24 hours. Groups were
compared by Student t-tests and the level of significance was set at p
Conclusions:
Maternal alcohol intake induced an anxiolytic-like effect and an impairment of spatial memory only in pups at PND60 that
disappeared at PND90, suggesting the involvement of neuronal plasticity mechanisms. The absence of any effect in pups at
PND30 might be due to the immature brains.
Keywords: anxiety, Elevated Plus Maze, ethanol, FAS, spatial memory
Financial Support: Fundao Padre Albino
Resumo:28-020
DNA DAMAGE AFTER THE ADMINISTRATION OF TECHNETIUM 99M - ECD IN RATS
Bortolotto, I. 1,2; Luiz, F. M. 1; Ramos, A. L. L. P. 1; Consiglio, A. R. 1,2

1
Dep Biofsica, Inst Biocincias, UFRGS
2
Programa de Ps Graduao em Enfermagem, UFRGS
Objectives:
The purpose of this study was to verify the presence of DNA lesion after the administration of Tc99m-ECD in rats, at doses
commonly used in human diagnostic nuclear medicine, adapted to the body weight of rats.

Adult male Wistar rats were divided into 3 groups, according to the intravenous treatment (Group 1: ethyl cysteinate dimer =
ECD, 200 L , n= 6; Group 2: 200 Ci Tc99m, n=6; Group 3: 200 Ci Tc99m-ECD, n=8). Twenty four hours after the drug
administration, the trunk blood, the brain areas (prefrontal cortex and hippocampus), and the bone marrow were extracted for
DNA analysis by the comet assay. The cells were classified according to the damage index (DI) based on the comet tail size. Data
was submitted to a 2-way analysis of variance and the Dunnets test. Twenty four hours after the infusion of Tc99m-ECD, DNA
damage index was greater at the bone marrow (DI= 76.75 + 21.00) when compared to the hippocampus (DI= 32.50 + 8.94,
p<0.05).
Conclusions:
DNA damage was produced 24h after the infusion of a radiopharmaceutical commonly used in nuclear medicine for evaluation of
the central nervous system, the Tc99m-ECD; although not detectable in blood, damage was found at the bone marrow, a very
radiosensitive tissue. On the other hand, it does not necessarily mean that it is a permanent damage.
Keywords: comet assay, DNA damage, nuclear medicine, radiobiological effects, technetium
Financial Support: Projeto Radioanlise n. 3504./FAURGS
Resumo:28-021
LOW-INTENSITY LASER INCREASES PLASMA PROTEINS AND INDUCES OXIDATIVE STRESS IN VITRO
Fonseca, A. S. 1,2,5; Presta, G. A. 1; Geller, M. 2,4; Paoli, F. 2,3; Valena, S. S. 4

1
Departamento de Cincias Fisiolgicas, Instituto Biomdico, UNIRIO
2
Centro de Cincias da Sade, UniFESO
3
Departamento de Morfologia, Instituto de Cincias Biolgicas, UFJF
4
Instituto de Cincias Biomdicas, Centro de Cincias da Sad, UFRJ
5
Depto. de Biofsica e Biometria, Instituto de Biologia, UERJ
Objectives:
Low-intensity laser therapy is based on the excitation of endogenous chromophores in biotissues and free radicals generation
could be involved in its biological effects. In this work the effects of low-intensity infrared laser on plasma protein content and
oxidative stress in blood from Wistar rats were studied.
Blood samples from Wistar rats were exposed to low-intensity infrared laser (830 nm) at continuous wave and pulsed (2.5, 250
and 2500 Hz) emission modes at different fluencies (1, 4 and 8 J/cm2). Plasma protein content, thiobarbituric acid-reactive
species (t-BARs) formation and myeloperoxidase (MP) were carried out to assess effects of laser irradiation on blood samples.At
the higher fluency, data obtained to plasma protein content were (XSD): 1.10.21 (control), 1.80.24 (continuous), 1.70.08
(2.5Hz), 1.80.41 (250Hz), 1.90.24 (2500Hz); to t-BARs: 0.10.05 (control), 0.20.22 (continuous), 0.20.08 (2.5Hz),
0.60.29 (250Hz), 0.60.23 (2500Hz); to MP: 0.70.12 (control), 0.30.11 (continuous), 1.20.37 (2.5Hz), 1.30.34 (250Hz),
0.50.22 (2500Hz). Low-intensity infrared laser exposure significantly (p
Conclusions:
Low-intensity infrared laser increases plasma proteins content and oxidative stress in blood samples suggesting that laser therapy
protocols should take into account fluencies, frequencies and wavelength of laser before beginning treatment.
Keywords: Blood, Laser, Oxidative stress, Plasma proteins, Wistar rats
Financial Support: FAPERJ, UniFESO, UERJ
Resumo:28-022
INFLUENCE OF GAALAS LASER ON SALIVARY FLOW IN HYPOTHYROID RATS
Jesus, V. C. D. 2; Santos, G. B. D. S. 2; Ramalho, M. J. P. 1; Ramalho, L. M. P. 2; Rodriguez, T. T. 1

1
Depto Biorregulao - Fisiologia, ICS-UFBA
2
Faculdade de Odontologia, UFBA
Objectives:
The salivary glands have a high rate of metabolism and blood flow; both are proportional to the rate of saliva formation and
indirectly dependant on thyroid function. The laser photobiomodulation (LPBM) acts as an agent stimulating salivation. This
study aims to investigate whether LPBM increased salivary flow rate on hypothyroid rats.
Male Wistar rats (250g), 07/group, were randomly distributed into two groups, euthyroid (EU) and hypothyroid (HYPO), treated
with propylthiouracil (PTU), 0.05g/100ml, administered orally for 4 weeks to induce hypothyroidism. Both groups had their
submandibular glands submitted to LPBM. Each group (EU and HYPO) was then divided into four subgroups according to the
protocol of irradiation: Control (without irradiation); Laser AsGaAl (Twin Flex): 660nm (40mW); 780nm (40mW) and
780nm (70mW). The dose results of 6 J/cm2 of energy on each gland, beam size (spot) of 0.4 cm, and total dose of 12 J/cm2 per
session. The first laser application occurred after 2 weeks of PTU induced, and repeatedly during 02 weeks, every 48 hours. On
the day of experiment, rats were anesthesized with ketamine (100mg/Kg) + xilazine (14mg/Kg), tracheostomized and stimulated
to salivate with the cholinergic agonist, pilocarpine i.p.(5mg/Kg body weight). The salivary flow (l/min/100g body weight) was
evaluated for 15 minutes from the time of the first drop of saliva. Hypothyroidism decreased salivary flow rate (control: 13,31
3,09; 660nm (40mW): 12,51 3,50; 780nm (40mW): 14,80 3,84; 780nm (70mW): 16,75 3,82; Mann-Whitney Test; P
Conclusions:
Our results suggest that LBPM dont stimulate salivary secretion in hypothyroid rats in the studied protocols.
Keywords: laser, hypothyrodism, saliva
Financial Support: FAPESB, CNPq, PIBIC/UFBA
Resumo:28-023
DESIGN OF APTAMER BASED RADIOPHARMACEUTICALS, PHOTOTOXIC AGENTS AND NANOPARTICLES

FOR TARGETED CANCER THERAPY
Missailidis, S. 1; da Pieve, C. 1; Makwana, V. 1; Antimisiaris, S. G. 2; Santos-oliveira, R. 3

1
Department of Chemistry and Analytical Sciences, The Open University
2
Pharmacy Department, University of Patras
3
Laboratory of Nanoradiopharmeceuticals, HUCFF
Objectives:
The aim of this project has been to use aptamers selected against the tumour marker MUC1 glycoprotein to engineer constructs
with photodynamic therapy agents, radiopharmaceuticals and nanoparticles that may provide novel treatments against breast and
other epithelial tumours.
Aptramers against the MUC1 mucin were selected using the SELEX methodology and analysed for their affinity and specificity
using ELISA, SPR, confocal microscopy and FACS. Aptamers coupled to quantum dots allow detection of circulating MUC1 as
a breast tumour marker. Light activated aptamers coupled to the heme like PDT agent chlorin e6 selectively kill epithelial cancer
cells. Similarly, Tc-99m labelled aptamers appear to localise on the tumour mass in experimental models in gamma-camera
imaging studies, whereas coupling of aptamers to linear and or combed PEG, carbon nanotubes, liposomes and silica mesoupors
changes their pharmacokinetic properties and yet allows them cancer cell binding and internalisation as well as delivery of
cytotoxic agents specifically to the cancer cell.
Conclusions:
Aptamers have been very promising in the development of advanced therapies, nanotechnology and targeted radiotherapy, as they
have the potential of confering specificity to phototoxic, radiopharmaceuticals and nanoparticles for cancer versus normal tissues,
thus increasing efficacy and minimising side effects and they are currently in development for clinical applications in the
treatment of cancer.
Keywords: aptamers, radiopharmaceuticals, phototoxic agents, nanoparticles
Financial Support: The Open University, Breast Cancer Campaign UK, Antisoma Limited, Euzoia Limited
Resumo:29-052
RAT LUMBRICAL MUSCLE: A NOVEL EX VIVO MODEL FOR EVALUATION OF ADULT SKELETAL MUSCLE
PROTEOLYSIS AND SCREENING OF DRUGS FOR ANTI-CATABOLIC PURPOSES
Bergantin, L. B. ; Figueiredo, L. B. ; Godinho, R. O.

Farmacologia, Unifesp EPM
Objectives:
Until now the pharmacological screening of drugs that interfere with skeletal muscle proteolysis have mainly been evaluated in
muscles from prepubertal rats (e.g., extensor digitorum longus - EDL and soleus), which are thin enough to allow adequate in
vitro diffusion of oxygen and substrates. However, the use of muscle at accelerated prepubertal growth has limited the analysis of
adult muscle. Thus, the effects of aging or neurodegenerative diseases have been restricted to measurement of static markers,
such as atrogin-1/MAFbx or Murf-1. In order to overcome this limitation, in the present study, we established a novel ex vivo
model to assess skeletal muscle proteolysis measuring the tyrosine release: the lumbrical muscles of rat hindlimb paw, which are
thin enough to allow plenty ex vivo oxygenation and metabolic perfusion for hours.
Lumbrical muscles (LB, 8/animal; 4/hindlimb paw) obtained from prepubertal, adult or senile rats were carefully dissected and
kept attached to their own metatarsal bones, by their tendons, at exactly resting length in order to avoid damaging. Each
muscle/bone system was transferred to tubes containing carbogen-saturated Tyrode solution, pH 7.4, plus cycloheximide (0.5
mM). After a 30 min equilibration period at 37C, tissues were incubated in 1.5 mL fresh medium of identical composition
drugs. The rate of proteolysis was determined by measuring the rate of tyrosine release in the incubation medium, using a
fluorometric method (at 485/590 nm) described by Waalkes and Udenfriend (J. Lab. Clin. Med., 50: 733, 1957) and adapted for
96-well microplate. By incubating lumbrical muscles (eight/ rat) attached to their individual metatarsal bones in Tyrode solution,
we showed that muscle proteolysis at adult and aged rats (3 to 24-month-old) was 45% to 25% of those from prepubertal animals
(30-day-old). While acute mechanical injury or 1-7 day denervation increased by up 60% tyrosine release from adult lumbrical
muscle, it was reduced by 20-28% after 2-h incubation of beta-adrenoceptor agonists, forskolin or phosphodiesterase inhibitor
IBMX. Using inhibitor of 26S proteasome (MG132) or the combination of calpain inhibitors (E64 plus leupeptin), we showed
that ubiquitin-proteasome is accountable for 40% total lumbrical proteolysis from adult rats.
Conclusions:
Our data show that this system can be used to evaluate skeletal muscle proteolysis in rats of different ages, and that protein
degradation decreases with aging, which may redefine nowadays concepts about skeletal muscle proteolysis. Besides, this system
allows the screening of 7 different doses of drugs at the same time by using a unique rat, differently from current methods, which
permit a single dose analysis. Thus, this system represents an important tool to allow high-throughput screening of drugs for
therapeutic anti-catabolic purposes.
Keywords: protein degradation, tyrosine release, aging, muscle, proteasome
Financial Support: CNPQ & FAPESP
Resumo:29-053
PROMISCUOUS COUPLING OF SKELETAL MUSCLE BETA2-ADRENOCEPTOR TO GS AND GI PROTEINS.
Rodrigues, F. S. M. ; Duarte, T. ; Godinho, R. O.

Dept of Pharmacology, Universidade Federal de So Paulo , (UNIFESP-EPM)
Objectives:
One of the most well known mechanisms that increase skeletal muscle contraction force relies on drugs that elevate cAMP
intracellular concentration. This can be achieved by activation of Gs protein coupled receptors (GsPCR), such as beta2adrenoceptors (beta2-AR), of adenylyl cyclase (AC) or inhibition of phosphodiesterase (PDE). Interestingly, in heart, beta-ARdependent inotropic effect was attenuated by promiscuous receptor coupling to Gi protein (Heubach et al., Molecular
Pharmacology, 65:1313, 2004). In the present study we evaluate the possible functional coupling of mouse diaphragm beta2-AR
to Gi-protein and its role in modulating skeletal muscle contraction.

Inotropic effects of beta-AR agonists isoproterenol (non-selective) or clenbuterol (beta2-selective) were evaluated by measuring
isometric contractions of mouse diaphragm induced by transmural electrical stimulation (0.1 Hz frequency, 2 ms duration, and
supra-maximal voltage, at 30 degrees Celsius). To assess the additional beta-AR coupling to Gi-protein, the effects of
isoproterenol and clenbuterol were analyzed in diaphragm strips pre-incubated for 1 h with 1 g/mL pertussis toxin (PTX).
Isoproterenol (3-1000 nM; n = 4-6) and clenbuterol (1-1000 nM; n = 3-5) induced a bell-shaped concentration-dependent increase
in muscle contraction that was maximal (40-50%) at 100 nM. Maximum agonist effect was observed within 30 min after the
onset of stimulation, which was followed by a descending phase of inotropic effects. PTX increased by 20%-40% the maximum
inotropic effect of isoproterenol and clenbuterol, while inhibited the delayed descending phase. Ethical committee 0011/08.
Conclusions:
These results indicate a dual coupling of beta2AR to Gs and Gi proteins in skeletal muscle. The late activation of Gi protein
signaling pathways, which results in attenuation cAMP production may provide a negative-feedback loop that limits GsPCR
response and possible deleterious effects of excessive contraction.
Keywords: SKELETAL MUSCLE, BETA-ADRENOCEPTOR, COUPLING, Gs, Gi
Financial Support: CAPES, FAPESP and CNPq
Resumo:29-054
ANALYSIS OF STRUCTURAL REQUIREMENTS OF THE RECEPTOR AT1 IN ACTIVATION OF AKT AND ERK
1/2 SIGNALING PATHWAYS: ROLE OF EGFR TRANSACTIVATION.
Machado, M. A. 1; Costa-neto, C. M. 1; Reis, R. I. 2,1

Depto. de Bioqumica/Faculdade de Medicina de Ribeiro Preto, USP-FMRP
2
Laboratrio de Rim e Hormnios/ Escola Paulista de Medicina, UNIFESP
Objectives:
The binding of the natural agonist Angiotensin II (AngII) to the AT1 receptor triggers a series of intracellular responses. At our
laboratory we generated two AT1 receptors mutants (P82A and P207A) that showed to be unable to evoke full G-protein
activation when stimulated with this agonist. Therefore, this study aimed to evaluate the role of ERGR transactivation in Akt and
ERK 1/2 signaling pathways in cells transfected with those receptor mutants comparing with the activation profile obtained in
cells transfected with the wild type AT1 receptor.
HEK293T cells were transiently transfected with the wild type and the mutant receptors. Cells were stimulated with AngII (1M)
after treatment with the angatonist for the EGFR (AG1478, 5M). Akt and ERK1/2 signaling pathways were analyzed by
Western Blotting using specific antibodies. As concerning to ERK1/2 activation, the mutant receptors showed a similar profile as
compared to the wild type receptor and the activation of this signaling pathway showed to be independent of EGFR
transactivation. Concerning to Akt activation, our results suggest that the P82A receptor mutant and the wild type AT1 receptor
are able to activate this signaling pathway independently of G-protein. In addition, after treatment with the EGFR antagonist we
observed a decrease in Akt phosphorylation evidencing a role of EGFR transactivation in cells transfected with those receptors.
Interestingly, the P207A mutant receptor showed no activation of Akt signaling pathway.
Conclusions:
Based on the above described results we can infer that the AT1 receptor can be stabilized in distinct conformations, which are
able to trigger different signaling pathways. Understanding the structural requirements of the AT1 receptor might contribute to
the future rational design of new drugs to this receptor.
Keywords: Activation of Akt, Activation of ERK1/2, Angiotensin II, AT1 receptor, EGFR transactivation
Financial Support: CNPq, FAPESP, FAEPA.
Resumo:29-055
DOPAMINE MODULATES ASCORBATE RELEASE: RECEPTORS AND PATHWAYS INVOLVED
Encarnao, T. G. ; Portugal, C. C. ; Paes-de-carvalho, R.

Department of Neurobiology/ Fluminense Federal University, UFF
Objectives:
Ascorbate (asc) is a neuromodulator present in the central nervous system (CNS), where it plays important physiological
functions. Dopamine (DA) is an important neurotransmitter that acts through receptors classified as D1-like, positively coupled to
adenylyl cyclase (AC), and D2-like, related to AC inhibition. Our objective in this work was to investigate if DA could modulate
asc release and to study the receptors and pathways involved in this function.
The release assay was performed using cultured retina cells from 8-day-old chicken embryos (E8) incubated with [14C] asc. We
observed that DA modulates asc release in a dose-dependent manner (EC50 of 293.7 nM and maximal release of 179.1 11.6 %,
n=3). Treatment with DA (50 M) stimulated asc release (71.1 18.3 %, n=3) and this effect could be observed after a second
treatment with DA (115.9 21.3%, n=3). The DA effect was not inhibited by Pargyline (50 M), a MAO (an enzyme that
degrades DA) inhibitor (DA: 66.1 8.1 %; n=19; DA + Pargyline: 54.8 10.3 %; n=3), but was inhibited by haloperidol (15 M)
(11.8 7.7 %, n=3), a DA receptor antagonist. SKF-38393 (10 M), a D1 agonist, stimulated asc release (52.9 7.8 %, n=14)
and Quinpirole (10 M), a D2 agonist, had no effect (-2.3 4.5 %, n=5). The SKF-38393 effect was inhibited by SCH-23390 (10
M), a D1 antagonist (16.9 7.9 %, n=3). Moreover, SKF-38393 and DA effects were inhibited by MDL-12,330 (10 M), an AC
inhibitor (22.5 6.8 %, n=3; and 15.5 3.6 %, n=3, respectively). The PKA inhibitors H-89 (5 M) or KT 5720 (1 M) were not
capable of blocking the effect of DA (73.2 21.1 %, n=5; and 79.6 10.9 %, n=3) or SKF-38393 (41.8 9.9 %, n=4).
Otherwise, the effect of Me-cAMP (100 M), an EPAC (other cAMP target) activator, resembled the DA effect (51.2 10.6 %,
n=2). We have also investigated the involvement of other protein kinases using Ly 294,002 (10 M), a PI3K inhibitor, as well as
PD 98,059 (10 M) or UO126 (10 M), MEK inhibitors, that blocked the DA effect (DA + Ly 11.3 11.1%, n=3; DA + PD 15.9
3.2 %, n=3; DA + UO126 15.7 11.3 %, n=2). Finally, we have also studied the participation of the asc transporter SVCT2
(Sodium-dependent Vitamin C Transporter 2) on the release stimulated by DA using the blockers Sufinpirazone (1 mM) or
Quercetin (500 M) as well as the effect of sodium absence. All these treatments completely inhibited the DA effect (DA +
Sufinpirazone: -59.4 5.7 %, n=6; DA + Quercetin: -12.2 5.6 %, n=2; DA without sodium: -12.6 5.2 %, n=4).
Conclusions:
Our results provide evidence that asc release in cultured retina cells is under DA control through the D1 / AC / cAMP pathway.
However, this effect is clearly PKA-independent but EPAC / PI3K / MEK-dependent, and probably involves SVCT2 reversal.
Keywords: ascorbate, dopamine, D1 receptor, retina, SVCT2
Financial Support: CNPq, CAPES, FAPERJ, PRONEX/MCT
Resumo:29-056
BRAIN MARKERS OF NEURODEGENERATION IN SEPSIS SURVIVORS RATS
Petronilho, F. 1,2; Miguel, S. P. 1; Danielski, L. G. 1; Guolo, K. 1; Pasquali, M. 3; Gelain, D. P. 3; Grunwald,

M. S. 3; Quevedo, J. 4; Moreira, J. C. F. 3; Dal-pizzol, F. 2
3
PPGCiencias Biolgicas Bioquimica, UFRGS
1
LIMEX/PPGCS, UNISUL
2
FISIPAT/PPGCS, UNESC
4
Neurolab/PPGCS, UNESC
Objectives:
Survivors from sepsis presented cognitive deficits that were associated with decrease quality of life increasing long-term
morbidity, and certain aspects of these alterations overlaps the pathophysiological mechanisms for neurodegeneration. In this
context, the receptor for advanced glication (RAGE) seemed to be an important step in neurodegeneration progression, thus we
here determined the presence of markers of neurodegeneration in brain of sepsis survivors rats and the possible involvement of
RAGE in these alterations.
Male Wistar rats (250-350g) n=10 per group ,were subjected to sepsis by cecal ligation and puncture and thirty days after animals
were killed, hippocampus and pre-frontal cortex isolated to the determination of beta-amyloid, alpha-synuclein and
phosphorilated tau and RAGE and the activation of its downstream signaling molecule ERK 1/2 content by western
blotting.Results: Beta-amyloid, alpha-synuclein and phosphorilated tau were increased in the hippocampus, but not in the prefrontal cortex. In the hippocampus increased the content of RAGE and the activation of ERK 1/2.
Conclusions:
Brain from sepsis survivors animals presented several markers of neurodegeneration suggesting that the RAGE-MAPK pathway
could be responsible to long-term cognitive deficits in sepsis survivors.
Keywords: sepsis, neurodegeneration, RAGE
Financial Support: INCT, CNPq, CAPES
Resumo:29-057
VASCULAR ACTIVITY AND EXPRESSION LEVEL OF KKS AND RAS RECEPTORS IN B1R OVEREXPRESSED
RAT AORTA
Silva, R. F. 1; Rodrigues, E. S. 1; Martin, R. P. 1; Oliveira, S. M. 1; Nakaie, C. R. 1; Merino, V. F. 1;

Pesquero, J. B. 1; Bader, M. 2; Shimuta, S. I. 1
1
Biofsica, UNIFESP
2
Max Delbrck Center for Molecular Medicine, MDC
Objectives:
Our aim was to investigate the vascular reactivity to the agents of Kalikrein Kinin System (KKS) and Renin Angiotensin System
(RAS) in the isolated thoracic aorta of rats overexpressing the kinin B1 receptor on the vascular endothelium (TGR(Tie2)). In
addition the expression of the B1R, B2R and AT1R receptors was assessed using real-time PCR technique.
Experiments were carried out using 300-350g Sprague-Dawley rats as control (WT) and overexpressing B1R (TGR(Tie2B1)).
For vascular reactivity, isometric relaxation and contraction responses to kinins and angiotensins curves (10-10 10-6M) were
recorded using a bath organ with PowerLab transducers and analyzed with LabChart Software. For receptors expression, the real
time PCR technique was performed with an ABI PRISM 7000. The relaxant effect of DBK (N=8) , as expected, was increased
in the transgenic rat, 73 5.5 % compared with the WT rat, 20 5.0%, but the apparent affinity (pD2, -log [EC50]) of the
receptor (N=8) remained unchanged, 8.6 0.3 (TGR(Tie2)) and 8.0 0.7 (WT). Unexpectedly, the responsiveness to BK (N+7)
was also elevated in the TGR(Tie2) rat, 54 6.0, while it was 21.5 4.0 in the WT rat. The apparent affinity of receptor B2R was
unaltered and the values of pD2 were: 7.9 0.4 (TGR(Tie2)) and 8.1 1.0 (WT) (n=7). The expression of kinin receptors (N=5)
were shown to be increased for both, B1R (7.3 folds) and for B2R (3.3 folds). In the RAS system no alteration were observed.
The values for maximum effect in comparison with KCl-induced effect and of pD2 for AngI were (N=6): 41 8.0% and 7.9 0.3
in the control rat, and 42 9.0% and 7.7 0.3 in the transgenic rat. For AngII the values for Emax and pD2 were (N=7): 32
12.0% and 7.6 0.2 in the TGR(Tie2), and 31 4.0% and 7.7 0.2. The expression of AT1R also showed no difference between
the control and transgenic rats (n=4).
Conclusions:
Our findings provided evidence that, expression of exogenous B1 receptor specifically in the vascular endothelium induced an
increase in the expression level of B2 receptors without affecting the AT1R expression. The data suggest the occurrence of
receptor autoregulation by kinins without cross-talking between KKS and RAS in the transgenic rat aorta overexpressing the B1
receptor.
Keywords: angiotensin, bradykinin, des-arg9-bradykinin, kinin receptors, real time PCR
Resumo:29-058
SIGNALING PATHWAYS ACTIVATED BY INTERLEUKIN 4 IN THE RETINA OF NEWBORNS RATS
Araujo-martins,l2; Santos, A. A. 2; Araujo, E. G. 1

Neurobiologia / Universidade Federal Fluminense, UFF
2
Fisiologia e Farmacologia / Universidade Federal Fluminense, UFF
1
Objectives:
Interleukin-4 is a pleiotropic cytokine and its effects are already well known in the immune system. Concerning the nervous
system, during the last decades, it was demonstrated that IL-4 plays a neuroprotective role in this system. Interleukin 4 is a small
protein (14,9 kDa) and its biological effects are mediated by IL-4 receptors. The aim of this study was to evaluate the signaling
pathways stimulated by the treatment with interleukin-4 (IL-4) in rat retinae. Literature shows that different signaling cascades
can be induced following IL-4 binding. It was already demonstrated that activation of IL-4 receptors stimulates at least 3 different
pathways: Jak-stat6, Raf-MEK-MAPK and PI3K. Moreover, the increase in intracellular calcium and cAMP levels as well as
PKC activation can also occur.
Lister Hooded rat pups (P0-P2) were killed by decapitation and their retinae were dissected in a saline salt solution. In our
experimental protocols we used only intact retinas. The treatment of retinal tissue was performed using complete medium (199,
5% FCS, glutamine and antibiotics) in the presence of IL-4 and or different drugs. Control retinae were maintained in complete
medium. During all the experiments the retinal tissue was incubated in atmosphere of 5% CO2 and 95% air, at 37C, for different
time intervals. To analyze protein expression and/or protein phosphorylation we used the Western blot technique. Our results
show that treatment of retinal tissue with IL-4 induces an increase in the pCREB; an effect inhibited by 1M H89 (an inhibitor of
PKA activation) [CT=100%, H-89=121.43% 11.44, IL-4= 233.19% 25.44, IL-4 + H89=158.84% 7.71 n=3]. We also
observed that the effect of IL-4 on CREB phosphorylation was blocked when the MEK pathway was inhibited by PD 98059
50M (CT= 100%, PD=111.67% 30.64, IL-4= 192.33% 2.49 and IL-4 + PD=155.67% 4.03 n=3). Our data shows that IL-4
treatment also increase BDNF expression and this effect depends on adenylyl cyclase activity (CT =100%, SQ (adenylyl cyclase
inhibitor)=128.44% 20.89, IL-4 =174.14% 8.81, IL-4 + SQ =143.18% 12.33 n=3) and PKA activity (CT =100%, H89=
110,67% 15,91, IL-4 =153,24% 21,20, IL-4 + H89= 113,17% 7,71 n=3).
Conclusions:
Our data allow us to suggest that the effect of IL-4 on BDNF expression is mediated by PKA and MEK pathways.
Keywords: INTERLEUKIN 4, RETINA, RATS, cAMP, BDNF
Financial Support: : CAPES, CNPq, FAPERJ and PRONEX
Resumo:29-059
MUTATION OF SER418 MODULATES THE EXPRESSION OF ENDOGENOUS HSGLT1.
Beloto-silva, O. ; Oliveira-souza, M.
Department of Physiology and Biophysics, ICB1
Objectives:
Glucose is the main energy source for the cells and the Na+/glucose co-transporter (SGLT) participates in the maintenance of
glucose homeostasis. It is known that protein kinase A (PKA) can phosphorylate the Serine 418 (S418) of the SGLT1 to change
its distribution in the cells. In this study was investigated, in Human Embryonic Kidney (HEK-293) cells, if the overexpression of
wild-type human SGLT1 (hSGLT1-WT) and the mutation of S418 to Histidine (hSGLT1-S418H) can modulate the expression of
endogenous hSGLT1 and the intracellular pH recovery rate (dpHi/dt).

The cDNA of hSGLT1-wt and hSGLT1-S418H were cloned into the expression vector pEGFP-N1, in the XhoI and HINDIII
sites. All the recombinants (wt and S418H) were transfected into HEK-293 cells and the stable transfection was confirmed by
Confocal microscopy. The HEK-293 cells not-transfected (NT), transfected with hSGLT1-WT (WT) or transfected with
hSGLT1-S418H (S418H) grown to confluence in DMEM medium containing glucose 5 mM. Western blot analysis confirmed
the expression of recombinants hSGLT1 WT or S418H and it was similar to endogenous SGLT1 expression. The pHi recovery
rate was analyzed by fluorescence microscopy using BCECF-AM fluorescent probe. The dpHi/dt did not change in WT
[0.3170.020 (n=7)] and S418H [0.2870.030 (n=7)] in comparison to the NT cells [0.2930.020 (n=7)].
Conclusions:
The preliminary results indicate that, to maintenance of glucose homeostasis, HEK-293 cells can modulated the endogenous pool
of the hSGLT1 when they are transfected with both hSGLT1-WT or S418H. Additionally, this factor does not permit
modifications in the dpHi/dt. However, these results will be confirmed with analysis of the Na+/H+ exchange (isoform 1 and 3 NHE1 and NHE3) expression. Previous data from our laboratory (J. Membr. Biol, 239(3):157-165, 2011), using HEK-293 cells,
demonstrated that the inhibition of PKA modifies the membrane expression of hSGLT1 and the dpHi/dt. Seemingly, in the same
cells, the mutation seems does not affect the hSGLT1 activity, indicating that this protein, in the presence of glucose 5 mM, can
be also modulated by other factors than PKA.
Keywords: Human SGLT1, Intracellular pH, Na+/H+ exchange
Resumo:29-060
INFLUENCE OF P2X7-GENE DELETION IN A MURINE MODEL OF MYCOBACTERIUM TUBERCULOSISINFECTION
Santos, Aa Jr3,4,1; Rodrigues-junior Vs3,5; Coutinho-silva R2; Santos, D. S. 3; Campos, M. M. 3,6,7; Morrone,
F. B. 1,4,6
1
Laboratrio de Farmacologia Aplicada - Faculdade de Farmcia, PUCRS
2
3
INCT-TB / CPBMF, PUCRS
4
Programa de Ps-Graduao em Biologia Celular e Molecular , PUCRS
5
Programa de Ps-Graduao em Medicina e Cincias da Sade , PUCRS
6
Instituto de Toxicologia, PUCRS
7
Faculdade de Odontologia, PUCRS
Objectives:
It was previously demonstrated that treatment of Bacillus Calmette-Gurin (BCG)-infected human macrophages with ATP
induced P2X7 receptor-mediated killing of intracellular mycobacteria (BMC Immunol. 9:35, 2008). Recently, we have
demonstrated that purinergic P2X7 receptor expression is found significantly augmented in the lungs of mice infected with
Mycobacterium tuberculosis. In the present study, we have further investigated the role of P2X7 receptors during in vivo M.
tuberculosis infection, by using knockout mice to P2X7 receptor.

Male P2X7 receptor knockout (P2X7(-/-)) and C57BL/6 wild type (WT) mice (8 per group, 25-30 g) were used. All the
experimental protocols were approved by the Local Animal Ethics Committee (CEUA 10/00203-PUCRS). The infection model
was accomplished according to the methodology described before (Antimicrob. Agents Chemother. 49:2816, 2005). The animals
were anesthetized and received an intravenous injection of 200 l of a M. tuberculosis suspension (H37Rv strain; 5 x 108
CFU/ml). Control mice received the same volume of saline. The procedures were carried out in a level III security cabinet. The
infection was confirmed by using the specific Ziehl-Neelsen staining. After 28 days of infection, the animals were euthanized.
Immediately after, the spleens and the right lungs were weighed and homogenized in 1 ml of 0.9 % saline solution. The
homogenates were serially diluted (10-fold scale), and cultured onto 7H10 agar plates. Colonies were counted after 4 weeks of
incubation. Tissue burden of M. tuberculosis was expressed as log10 CFU per organ. Infected WT mice showed a marked
increase in the spleen weight, in comparison to non-infected animals, indicating the occurrence of splenomegaly. Notably, the
spleen weight in M. tuberculosis-infected P2X7(-/-) mice was significantly higher (3.78 0.48 mg/g body weight), when
compared to WT infected mice (2.55 0.18 mg/g body weight) (PM. tuberculosis-infected P2X7(-/-) mice showed an increase of
0.881 log10 CFU/lung tissue, in relation to infected WT mice (P<0.0001).
Conclusions:
The augmentation of lung burden of M. tuberculosis and the increase of spleen weight observed in M. tuberculosis-infected
P2X7(-/-) mice suggest that this receptor have a key role for understanding the pathogenesis of tuberculosis.
Keywords: Mycobacterium tuberculosis, P2X7, P2RX7, Tuberculosis
Financial Support: CNPq-INCT-TB, CAPES, BNDES, PUCRS.
Resumo:29-061
A PUTATIVE RECEPTOR TYPE FOR CRUSTACEAN RED PIGMENT CONCENTRATING HORMONE.
Milograna, S. R. ; Mcnamara, J. C.
Departamento de Biologia/FFCLRP, USP
Objectives:
Crustaceans exhibit a remarkable capability to rapidly change color, an effect due to the differential translocation of pigment
granules within the cytoplasm of their specialized effector cells, the chromatophores. The receptor type that binds the pigment
regulating neurosecretory peptide, red pigment concentrating hormone (RPCH), and triggers pigment aggregation in shrimp
chromatophores via the Ca2+ and cyclic GMP signaling cascades, has not been investigated. To explore the receptor type to which
RPCH binds at the chromatophore plasma membrane, we employed a Protein G antagonist (PGAnt), a peptide inhibitor of Gprotein coupled receptors; a non-NMDA glutamate receptor agonist (AMPA); and a non-NMDA glutamate receptor antagonist
(CNQX).
Female Macrobrachium olfersi were collected from the Paba River, near So Sebastio, on the northern coast of So Paulo
State. Red ovarian chromatophores were perfused in vitro with physiological saline (NCS, 360 mOsm/kg, pH=7.4, Na + 180, K+ 5,
Ca2+ 5.5, Mg2+ 1, HEPES 5, glucose 2, and NaH2CO3 2.5 mM) and their maximum diameters were measured every 2 min and
converted to mean ( SEM) maximum dispersion (%) and translocation velocity (m/min). Experiments consisted of (i) control
preparations (N=12) NCS (10 min), NCS+RPCH 5 nM (30 min) and NCS (30 min); (ii) PGAnt (N=7) NCS (10 min),
NCS+PGAnt 5 M (30 min), NCS+PGAnt+RPCH (30 min), NCS+PGAnt (30 min); (iii) AMPA (N=8) NCS (10 min),
NCS+AMPA 30 M (30 min), NCS+AMPA+RPCH (30 min) and NCS+AMPA (30 min); (iv) CNQX (N=7) NCS (10 min),
NCS+CNQX 50 M (30 min), NCS+CNQX+RPCH (30 min) and NCS+CNQX (30 min). Results: PGAnt alone has no effect but
inhibits RPCH-triggered pigment aggregation by 50%; this aggregation lasts only 12 min (maximum velocity 13 2 m/min
similar to RPCH-control 17 2 m/min, P=0.85), followed by spontaneous dispersion to 60 11%. RPCH washout leads to 93
5% dispersion. AMPA and CNQX alone induce pigment hyperdispersion to 114 4% and 107 3%, respectively, after 30 min
perfusion. AMPA+RPCH allows nearly complete pigment aggregation (5 2%; maximum velocity 16 2 m/min similar to
RPCH-control, P=0.86). RPCH washout leads to 95 5% dispersion. CNQX subtly inhibits the aggregating response to RPCH,
reaching 25 7% after 30 min (maximum velocity 16 2 m/min similar to RPCH-control, P=0.95). RPCH washout leads to
82.2 13.0% dispersion.
Conclusions:
The RPCH receptor appears to be of the G-protein coupled receptor type, since inhibition by PGAnt not only strongly diminishes
the pigment aggregating response but also stimulates precocious, spontaneous pigment dispersion. Further, neither inhibition nor
stimulation by non-NMDA glutamate receptor ant/agonists affect RPCH response, ruling out this receptor type. This is the first
evidence that RPCH acts via a plasma membrane-located, G-protein coupled receptor.
Keywords: GPCR, chromatophores, pigment translocation, signal transduction, shrimp
Financial Support: CAPES, CNPq, FAPESP 08/52647-0; CEBIMar 2005/13.
Resumo:29-062
IDENTIFICATION OF NEW PUTATIVE PRPC LIGANDS BY PHAGE DISPLAY
Americo, T. A. 1; Carneiro, M. V. 1; Magdesian, M. H. 2; Linden, R. 1

1
IBCCF/UFRJ, IBCCF
2
IBqMed/UFRJ, IBqMed/UFRJ
Objectives:
Prion protein (PrPC) is a sialoglycoprotein anchored to the external face of the citoplasmic membrane by
glycosylphosphatidylinositol. Although related to diverse functions such as cellular adhesion, proliferation and cellular
differentiation, apoptosis and Cu+ and Zn+ transport and metabolism, its physiological role is still under discussion. Several
PrPC ligands were described, including the co-chaperone hop/STI-1. The interaction between these two proteins triggers various
signaling pathways such as AMPc/PKA, ERK and PI3K. The AMPc/PKA pathway is related to neuroprotection and the ERK
pathway to neuritogenesis and proliferation. It is not clear how PrPC, a protein without a transmembrane domain, can transmit
signals to the intracellular compartment. We used a phage-display approach to identify new PrPC ligands, presumably involved
in a signaling plataform with the hop/STI-1 protein.
A random heptapeptide phage display library was used to isolate clones that bind to PrPC after five biopanning rounds. The
binding specificity was tested by ELISA. The selected clones (20) were sequenced and the predicted peptides sequences were
submitted to similarity searches against the non-redundant protein sequences database (NCBI) using the BLAST algorithm.
Conclusions:
A total of 327 protein hits were recovered from this analysis One of the clones showed similarity to an extracellular region of
Metabotropic Glutamatergic Receptors 1 and 5 (mGluR1/5). A functional interaction between mGluR1/5 and PrPC has been
validated (Beraldo et al, FASEB J 2011 Jan;25(1):265-79). However, the putative PrPC-binding domain is located next to a
putative hop/STI-1 binding site in the mGluR1/5 molecules. The functional interaction among hop/STI-1, PrPC and mGluR1
suggested by the data has yet to be validated.
Keywords: prion, phage display, mGluR1
Financial Support: CAPES, CNPq, FAPERJ and FAPESP
Resumo:29-063
EVALUATION OF TOLL-LIKE RECEPTORS SIGNALING IN THE RAT PINEAL GLAND: REGULATORY
NETWORK FOR MODULATING MELATONIN SYNTHESIS
da Silveira Cruz Machado, S. 1; Tamura, E. K. 1; Pinato, L. 1,2; Markus, R. P. 1

1
Departamento de Fisiologia/Instituto de Biocincias - USP, IBUSP
2
Departamento de Fonoaudiologia - Uni. Estadual Paulista, UNESP
Objectives:
The pineal gland, a circumventricular organ, plays a pleiotropic role both in the circadian system and in inflammatory responses
(Neuroimmunomodulation, 14: 126, 2007). In fact, we suggested that the suppression of nocturnal melatonin production play a
role in the endothelial adhesion molecules expression, presumably allowing cell migration (Plos One, 5:e13958). We also have
shown that the rat pineal gland express proteins for recognition of pathogen- and danger-associated molecular patterns (PAMPs
and DAMPs) as CD14 and Toll-like receptor 4 (TLR4). These receptors induces the nuclear translocation of p50/p50 and
p50/RelA nuclear factor kappa B (NFKB) dimers triggering the inhibition of melatonin and N-acetylserotonin synthesis and the
induction of TNF production in cultured glands, respectively (J Pineal Res, 49:183, 2010). From the observations obtained so far,
there are prospects to assess whether the pineal gland presents the repertoire to recognize other PAMPs and DAMPs besides
lipopolysaccharide. Here we aimed to disclose the transcription of several genes involved in the recognition and in the
downstream signaling of PAMPs and DAMPs in the rat pineal gland.
Approved ethical committee (CEA/IB: 045/07). Male wistar rats kept in light/dark cycle (12:12 h) were killed between 4h and 5h
after lights on. Fresh-obtained rat pineal glands were collected, followed by mRNA extraction, cDNA synthesis and real-time
RT-PCR was performed using iCycler equipment. The reactions were carried out in order to analyze the expression of 84 genes
using a Toll-like receptors signaling pathway array kit (SABiosciences, Frederick, MD, USA) according manufactures
instruction. We detected the constitutive presence of all TLR mRNA analyzed (TLR from 1 to 9), except for Tlr5. We also
detected the constitutive gene transcription of the cytokine receptors Il1r1, Il6ra and Tnfrsf1a, for members of NFKB, Interferon
Regulatory Factor, JNK/p38 and JAK/STAT downstream signaling, as well as for effectors, adaptors and for TLR interacting
proteins both related to MYD88, TRAM, TIRAP and TRIF signaling. We also observed constitutive transcription for cytokines
and chemokines.
Conclusions:
The p50/p50 NFKB dimer is constitutively activated in a circadian manner in the rat pineal gland (Chronobiol Int, 27: 52, 2010).
Here we observed a constitutive transcription of the 5 subunits of this family. In addition we detected constitutive expression of
TLRs family genes, known to trigger different NFKB dimers nuclear translocation. As observed by TLR4 activation, the
activated NFKB pathway in the rat pineal gland modulates both melatonin synthesis and cytokine production (J Pineal Res,
49:183, 2010). Moreover, clinical and experimental data indicates a modulation in the melatonin production during several
disorders, such as neurodegenerative, diabetes, migraine, carcinosarcoma, heart stroke and local or systemic inflammation. As
TLRs detect endogenous and/or foreign molecules, the presence of these receptors in the pineal gland points to a constitutive
repertoire for modulating melatonin synthesis, necessary for allowing cell migration and the mounting of inflammatory
responses. In such context, here we provide molecular evidences to include the pineal gland as a sensor and player in the
inflammation.
Keywords: Pineal gland, Toll-like receptors, immune-pineal axis, NFKB signaling network, Inflammation
Resumo:29-064
ESTROGEN RECEPTORS IN PC-3 PROSTATE CANCER CELLS
Pisolato, R. ; Lucas, T. F. G. ; Lazari, M. F. M. ; Porto, C. S.

Depto. de Farmacologia/Universidade Federal de So Paulo, UNIFESP
Objectives:
Estrogen plays an important role in development and homeostasis of prostate tissue and etiology of prostatic diseases. The direct
effects of estrogen on the prostate are mediated by the classical estrogen receptors ESR1 and ESR2 that are localized primarily in
stromal and epithelial cells, respectively, and have different action profiles (Steroids 73:233, 2008). The expression of these
classical receptors in the androgen-independent prostate cancer cell line PC-3 has been conflicting (Cancer Res. 60:3175, 2000;
Eur. Urol. 40:557, 2001; Prostate 55:180, 2003), therefore, their expression needs to be further characterized. The G protein
coupled estrogen receptor (GPER), which mediates rapid (nongenomic) signaling of estrogen, is also described in PC-3 cells
(Cell Death Differ.17:1511, 2010). Recently it was identified and cloned a novel 36 kDa isoform of the full-length ESR1
designated ESR1-36 that is primarily localized in the cytoplasm and the plasma membrane and responds to membrane-initiated
estrogen and antiestrogen signaling pathways in breast cancer cells, for example, in MCF-7 cells (Biochem. Biophys. Res.
Commun. 336:1023, 2005; Proc. Natl. Acad. Sci. 103:9063, 2006). The presence of ESR1-36 has not been described in prostate
yet. This study was performed to characterize the expression of the classical estrogen receptors (ESR1 and ESR2), GPER and
ESR1-36 in PC-3 cells.
PC-3 cells were grown in RPMI medium without phenol red containing 0.02 mg/ml of gentamicin at 37C in a humidified
atmosphere with 5%CO2. Conventional RT-PCR was performed for detection of GPER and ESR1-36. Transcripts of the
expected size were detected in PC-3 cells and their identity will be confirmed by automated sequencing. Immunofluorescence and
Western Blot assays were also performed, as previously described (Biol. Reprod. 78:101, 2008; Biol. Reprod. 83:307, 2010). The
expression of ESR1, ESR2 and GPER was confirmed in PC-3 cells. Furthermore, ESR1-36 is also present in these cells and may
play a role on estrogen actions.
Conclusions:
These results indicate the presence of different receptor targets for the estrogen action in prostate cancer cells. Therefore different
signaling pathways might be expected upon estrogen stimulation of these receptors.
Keywords: ESR1-36, estrogen receptors, GPER, PC-3, prostate cancer
Financial Support: Supported by FAPESP.
Resumo:29-065
SYNTHESIS OF INTERACTING CAR AND MAGI-1 DOMAINS.
Mateus, R. S. 3,4; Kolawole, A. O. 4; Yan, R. 4; Sharma, P. 4; Hostetler, H. 4; Fiorino, P. 3; Farah, V. 3;

Excoffon, K. J. D. A. 4
3
Mackenzie Presbiteryan University, UPM
4
Wright State University, WSU
Objectives:
Adenovirus is both a respiratory pathogen and a candidate viral vector for gene therapy. The Coxsackievirus and adenovirus
receptor (CAR) is important for viral binding and entry into cells and hence CAR protein abundance and localization are
important for adenovirus infection. CAR has 2 different isoforms: CAREx7, which localizes to the basolateral (inside) surface of
polarized cells where it is inaccessible for viral infection; and CAREx8, recently discovered to localize to the apical membrane of
polarized primary human airway epithelia, where it can mediate initiation of adenovirus infection from the air exposed surface.
The two isoforms differ only at the intracellular C-terminus. It is known that MAGI-1b, a large cellular scaffolding protein with
many protein interaction domains, interacts with both isoforms in a PDZ dependent manner. Whereas, CAREx7 brings MAGI-1b
to the junctions, MAGI-1b causes the loss of CAREx8. We hypothesized that each CAR isoform interacts with different MAGI-1
PDZ-binding domains (PDZ0-PDZ5). To investigate this, we cloned each MAGI-1 PDZ domain and each CAR C-termini into
bacterial expression vectors and purified the proteins in order to study the interactions in vitro.
To determine the specific PDZ domain(s) that bind to each CAR isoform, fusion proteins for N-terminally His-GST-tagged
CAREx7-c-terminus (aa 261365), CAREx8-c-terminus (aa 261-352), and individual MAGI-1 PDZ domains (aa 20-110, 465555, 630-730, 840-930, 990-1080, 1140-1230) were prepared by cloning PCR fragments of CAR and MAGI-1 into the vector
pHH2. Clones were verified by PCR, restriction digest, and DNA sequencing. Plasmids were then transformed into Rosetta2
competent cells (EMD). Protein synthesis was induced with IPTG during 4 hours of incubation. The protein was extracted from
the culture pellet using GST-Bind Fractogel Cartridges (EMD), and the His-GST tags were removed with PreScission Protease
enzyme (GE Healthcare). Purified proteins were confirmed on 10% SDS-PAGE, followed by Coomassie blue staining, distaining
in 30% methanol, and demonstrated protein bands at the expected size of between 30kDa and 46kDa.
Conclusions:
CAR C-termini and MAGI-1 PDZ domains were successfully cloned and purified. The proteins will be used to study the
interaction between MAGI and CAREx7 and CAREx8 by in vitro Fluorescence Resonance Energy Transfer (FRET).
Keywords: adenovirus infection, Coxsackievirus and adenovirus receptor, MAGI-1, protein interaction, gene therapy
Financial Support: CAPES/FIPSE (R.S.M.), NIH 1R15AI090625-01 (K.E.), WSU BARE (A.K.).
Resumo:29-066
CROSSTALK BETWEEN ALBUMIN AND RENIN-ANGIOTENSIN SYSTEM IN RENAL SODIUM EXCRETION
Lopes, J. V. 1,2; Perucheti, D. B. 1,2; Landgraf, S. S. 1,2; Takiya, C. M. 2,1; Caruso-neves1,2

1
Instituto de Biofsica Carlos Chagas Filho/UFRJ, IBCCF/UFRJ
2
Instituto de Cincias Biomdicas/UFRJ, ICB/UFRJ
Objectives:
Recently, we observed in our group that albumin modulates the proximal tubule (PT) (Na+K+)ATPase in low concentrations.
Albumin also increases angiotensin II (Ang II) in PT, which is involved in pro inflammatory effects of the overload of albumin in
PT. Furthermore, Ang II can modulate the uptake of albumin, also the (Na+K+)ATPase activity in PT. Thus, our question is: Can
albumin modulate the (Na+K+)ATPase through Ang II signaling? The objective of this work was to verify the existence of a
crosstalk between albumin and Ang II on the PT (Na+K+)ATPase modulation
In vitro model - LLC-PK1 cells, a well characterized porcine proximal tubule cell line, were used. These cells were cultured in
DMEM with 10% FBS, 1% penicillin and streptomicin (37C and 5% CO2). When achieved the confluence, the cells were
starved and incubated in different conditions; In vivo model it was used acute renal failure (ARF) animal model, that was
induced in 14 weeks old male Wistar rats with intraperitoneal injection of 10g/kg/day bovine serum albumin during 10 days. The
different groups were treated or not with 30mg/kg/day losartan, an AT1 receptor antagonist, by gavage. In LLC-PK1 cells, we
observed that 0.01 mg/mL albumin increased the (Na+K+)ATPase activity (90%), while this effect was lost with higher albumin
concentration (20.0 mg/mL albumin). 10-6M losartan abolished the stimulatory effect of 0.01 mg/mL albumin while 10-8M
PD123319, an AT2 receptor antagonist, did not change it. Furthermore, 10-6M losartan abolished the inhibitory effect of higher
albumin concentration (20.0 mg/mL). In vivo experiment revealed that losartan treatment did not reverse the proteinuria,
interstitial collagen deposition and glomerular lesion observed in ARF model group. On the other hand, the increase of renal
sodium fraction excretion (FENa+) observed in ARF model was abolished with losartan treatment. In renal cortex of ARF group
was observed: (1) decrease of (Na+K+)ATPase activity and expression (65%); (2) decrease of AT1 expression (34%) and
increase of AT2 expression (100%). However, the losartan treatment in the ARF group reverts: (1) (Na+K+)ATPase activity and
expression; (2) the increase of AT2 expression. The ratio AT1/AT2 was decreased in ARF model and losartan-treated ARF group
partially restored this balance.
Conclusions:
Our data demonstrate that the effect of albumin on the PT (Na+K+)ATPase is mediated by Angiotensin II/AT1 receptor.
Keywords: albumin, biofsifa renal, caruso-neves, renal sodium excretion, renin-angiotensin system
Financial Support: FAPERJ, CAPES, CNPq
Resumo:29-067
THE MECHANISM OF THE FACILITATORY EFFECT-INDUCED BY METHYLPREDNISOLONE ON SKELETAL
NEUROMUSCULAR TRANSMISSION: THE INVOLVEMENT OF EXCITATORY M1 AND INIBITORY M2
MUSCARINIC AUTORECEPTORS.
Ambiel, C. R. 1; Dal Belo, C. A. . 2; Corrado, A. P. 3; Alves-do-prado, W. 1

1
Universidade Estadual de Maring, UEM
2
Universidade Federal do Pampa, UNIPAMPA
Faculdade de Medicina de Ribeiro Preto, FMRP
Objectives:
We have previously demonstrated, using mouse diaphragm preparations, that the methylprednisolone benefit of impaired
neuromuscular junctions may involve the increase of spontaneous acetylcholine release (Muscle & Nerve 26: 37, 2002). The aim
of this study was to investigate the influence of cholinergic autoreceptor in the methylprednisolone-induced facilitatory effect
using rat phrenic nerve-diaphragm preparations (RPND) direct and indirect stimulated.
The RPND were mounted essentially as described elsewhere (Brit. J. Pharmacol. 1, 38, 1946). The ratio (R) values was obtained
between the muscle tensions registered at the end (B) and at the beginning (A) of high-frequency (50 Hz, 5 sec) elicited by
electric indirect stimulation (R=B/A). R values were measured followed a previous 35 min methylprednisolone incubation, in
absence or presence of antimuscarinic agents. The experimental procedures were approved by The Ethics Committee in Animal
Experimentation State University of Maringa. Methylprednisolone (0.3 mM) increased significantly the R-values when RPND
were indirectly (16.37.3%) or directly (curarized preparations) stimulated (n=4, p<0.05).
Conclusions:
The data highly suggest that the molecular mechanism involving the methylprednisolone facilitatory actions at neuromuscular
junctions is the M1 and M2 muscarinic receptors. We suggest that the cross-talking signaling between the M2 followed a
previous activation of M1 receptor may lead to a restriction of the corticoid facilitatory action. The results also support the idea of
combining atropine with methylprednisolone in order to improve neuromuscular disorders therapeutics.
Keywords: METHYLPREDNISOLONE, MUSCARINIC AUTORECEPTORS, NEUROMUSCULAR TRANSMISSION
Financial Support: CNPq, Fundao Araucria, CESUMAR, FADEC-UEM.
Resumo:29-068
EFFECT OF BUFADIENOLIDES IN NEW INTRACELLULAR SIGNALING PATHWAYS MEDIATED BY THE
NA+/K+-ATPASE
Amaral, L. S. ; Curcio, B. C. ; Touza, N. A. ; Cunha-filho, G. A. ; Silva, C. L. M. ; Nel, F. G. ; Quintas, L.

E. M.
Instituto de Cincias Biolgicas / UFRJ, UFRJ
Objectives:
The Na+/K+-ATPase is a transmembrane enzyme that actively transports sodium and potassium ions against their concentration
gradients and this function is specific inhibited by cardiotonic steroids (cardenolides and bufadienolides). These inhibitors also
promote the activation of signaling pathways via Na+/K+-ATPase through protein-protein interactions. One of these pathways
involves activation of the cascade Ras/Raf/MEK/ERK. The ERK protein participates in many cellular processes like cell
proliferation. Ouabain, a cardenolide now considered in mammals a steroid hormone, inhibits ion transport and also stimulates
ERK. How to dissociate between these two cellular effects is still unknown, but this would allow us to establish the structural
requirements of potential selective modulators for each of these two pathways. We evaluated the ability of natural bufadienolides
to activate the signaling cascade mediated by Na+/K+-ATPase through activation of ERK, and compare their ability to inhibit
Na+/K+-ATPase activity. We also evaluated the effect of bufadienolides in cell proliferation and viability.
LLC-PK1 cells (porcine proximal renal tubule) were treated 15 min with 1nM-10&muM bufalin (BFL), telocinobufagin (TCB),
marinobufagin (MBG), lysed with RIPA buffer, centrifuged at 13,000g for 15 min and the supernatants were used in Western blot
for evaluation of activation (phosphorylation) of MAP kinases ERK1/2. To evaluate the ability of Na+/K+-ATPase inhibition,
inhibition curves were performed with increasing bufadienolides concentrations in purified membrane preparations of pig kidneys
and analyzed by the colorimetric method of Fiske and Subbarow. The evaluation of cell proliferation was measured by counting
the number of Trypan blue-viable LLC-PK1 cells treated bufadienolides for 24, 48 and 72 h. Cell viability was measured by
detection of anti- (Bcl-2) and pro-apoptotic (Bax) proteins by Western blot analysis. For comparison, ouabain was used as
standard.TCB and BFL inhibited Na+/K+-ATPasic activity in a concentration-dependent manner in the same range of inhibition
achieved by ouabain (IC50 = 0.2 0.1 M TCB and 0.3 0.06 M BFL). MBG inhibited with a 10-fold higher IC50 (3.4 0.2
M), being less potent. Tests for evaluating phospho-ERK1/2 show that the pathway induction is achieved in concentrations
where no significant inhibition of enzyme activity is observed (i.e., significant stimulation at 1 nM). MBG, like ouabain, induced
significant cell proliferation after 48 (100 nM) and 72 h (1, 10 and 100 nM) of treatment, similar to ouabain. TCB and BFL
however had no effect on cell proliferation (1 and 10 nM) and even inhibited it at 100 nM. Preliminary data point out that while
the expression of Bcl-2 is increased, Bax is decreased in MBG- and ouabain-treated cells.
Conclusions:
These results indicate that bufadienolides present functional selectivity: (1) activating ERK1/2 at concentrations that do not affect
Na+/K+-ATPase activity; (2) promoting divergent effects despite acting on the same receptor.
Keywords: Bufadienolides, ERK, NA+/K+-ATPASE, Ouabain, Cell proliferation
Financial Support: CAPES, Faperj, CNPq
Resumo:29-069
MODULATION OF NEURONAL SURVIVAL IN RETINAL CULTURES BY VITAMIN C: ROLE OF GLIAL CELLS
AND CREB
Domith, I. C. L. ; Portugal, C. C. ; Socodato, R. ; Paes-de-carvalho, R.

Departamento de Neurobiologia, UFF
Objectives:
The retina is the tissue responsible in transduce the light information from the environment to the brain and is considered part of
the CNS due to its anterior neural tube direct derivation. The retina is an excellent CNS model because most neurotransmitters
and neuromodulators found in CNS are also found in retina. Retinal cultures are widely used because most neurochemical
properties of the in vivo tissue are maintained in vitro. Vitamin C is an important antioxidant found in the CNS. Its reduced form,
ascorbate (asc), has neuroprotective effects, while its oxidized form, dehydroascorbate, may generate cellular damage. The
mechanisms of oxidative stress induced by vitamin C have not been well elucidated. CREB (a protein that binds to CRE, the
cAMP responsive element) is a transcription factor of the leucine zipper type, capable of regulating the transcription of several
genes. PKA is able to phosphorylate CREB in its serine 133 residue, thus recruiting the co-activator CBP (a protein that binds to
CREB), a histone acetyl transferase protein. CREB is able to constitutively bind to CRE sequences located in promoter regions of
genes, thus showing that the transcription of several genes would be under its regulation. Our objective in this work was to study
the modulation of cell survival and phospho-CREB (pCREB) levels by asc in cultures of chick embryo retinal cells.
Cultures of 8-day-old chick embryo retinal cells were used for experiments after the fourth day of culture. Cell death was
measured by MTT assays and proteins were analysed by western blot. Different types of cultures were treated with asc at several
concentrations. We have observed an extensive cell death when purified neuronal cultures were treated with low doses of asc (30
M) (48.1 8.3%, n=3). However, in mixed cultures containing neurons and glial cells, we found that higher doses of asc (1.5
mM) were necessary to achieve similar cell death levels. Treatment with asc (1.5 mM) increased pCREB levels after 5 minutes of
incubation (175.5 14.0%, n=10) and stabilized after 30 minutes (207.8 30.1%, n=17) up to 60 minutes. Interestingly,
adenosine A2a, dopamine D1 and NMDA receptor antagonists were all able to inhibit this effect on pCREB.
Conclusions:
Our results highlight the importance of glia in the modulation of neuronal survival by asc and show that adenosine A2a,
dopamine D1 and glutamate NMDA receptors participate in the stimulation of CREB phosphorylation after acute treatment with
asc.
Keywords: Vitamin C, CREB, Avian retina
Financial Support: CAPES, CNPQ, FAPERJ and PRONEX/MCT
Resumo:29-070
IL-6 EFFECT ON SURVIVAL OF RETINAL GANGLION CELL: INVOLVEMENT OF A2A ADENOSINE
RECEPTOR
Ritt, K. 1,2; Pergolo-vicente, R. 1; Pereira, M. R. 1; Paes-de-carvalho, R. 1; Araujo, E. G. 1

1
BIOLOGIA/UNIVERSIDADE FEDERAL FLUMINENSE, UFF
2
CENTRO UNIVERSITRIO PLNIO LEITE, UNIPLI
Objectives:
The aim of this study was to evaluate the involvement of the adenosine A2 receptor in the trophic effect mediated by IL-6 on
retinal ganglion cells kept in culture for 48h.
Methods: Within the first 24h after birth Lister Hooded rats were anesthetized by hypothermia. One L of a solution of 10% HRP
in 2% DMSO was then injected into each superior colliculus. After 16h of HRP injections animals were killed, their retinas
dissected, treated with 0.1% trypsin and dissociated by trituration. Cells were plated in coverslips and kept in complete 199
medium for 4h or 48h at 37C in an atmosphere of 5% CO2/95% air. The presence of HRP in the cytoplasm of RGC was revealed
according to the protocol of Mesulan. Cells were fixed with a mixture of aldehydes in sodium phosphate buffer (0,1M), the
coverslips were washed in phosphate buffer and reacted with tetramethylbenzidine. Results: Initially we investigated if the IL-6
effect was mediated by adenosine transporter. To address this question we treated the cultures with NBI 10nM (Adenosine
transporter inhibitor) and we observed a total inhibition of the IL-6 effect when NBI was present [CT 4h: 100%; CT 48h:
59,023,05; IL-6 50ng/mL: 97,31,66; NBI 10nM: 58,143,93; NBI/IL-6: 49,520,5; n=4-5]. Following we investigate the
involvement of cAMP in the IL-6 effect. Using SQ22536 2M (Adenylyl Cyclase inhibitor) we obtained a blockade of the IL-6
effect [CT 4h: 100%; CT 48h: 55,3163,15%; SQ: 57,843,6%; IL-6: 96,782,53%; IL-6/SQ: 63,674,0%; n=5-10]. The next
step was to evaluate the participation of A2 adenosine receptor on IL-6 effect. We obtained an inhibitory effect after ZM 10nM
(A2 adenosine receptors blocker) [CT 4h: 100%; CT 48h: 52,061,44%; Zm: 51,651,57%; IL-6: 92,053,74%; IL-6/Zm:
53,380,98; n=6-12]. Corroborating these data treatment with CGS (A2 adenosine receptor agonist) induced a trophic effect on
RGC [CT 4h: 100%; CT 48h: 72,92,7%; CGS 1nM: 80,062,7%; CGS 5nM: 89,013,67%; CGS 10nM: 96,443,1%; n=5-6]
and the effect of CGS was mediated by cAMP [CT 4h: 100%; CT 48h: 66,312,91%; SQ: 61,221,27; CGS: 87,603,66%;
CGS+SQ: 61,732,26%; n=6-12]. We also evaluated the effect of the PKA inhibitor (H89 1M) on the IL-6 effect [CT 4h:
100%; CT 48h: 50,640,78%; H89: 48,145,79%; IL-6: 80,969,7%; IL-6/H89: 79,429,6%; n=6-10]. The results show that IL6 effect does not increase the survival of RGC via PKA activation.
Conclusions:
Our data indicate a participation of A2 adenosine receptor (A2R) on the trophic effect of IL-6 on retinal ganglion cell survival.
We can suggest a possible role of IL-6 controlling neuronal cells survival.
Keywords: Interleukin-6, A2 Adenosine Receptor, retinal ganglion cell
Financial Support: CAPES, CNPq, FAPERJ e PRONEX.
Resumo:29-071
EFFECTS OF LASSBIO-767 ON M2 AND M3 MUSCARINIC RECEPTORS AND HIPPOCAMPAL GABAERGIC
TONIC CURRENT.
Vieira, K. S. T. 1; Gamba, N. F. 1; Fraga, C. A. M. 3; Barreiro, E. J. D. L. 3; Bolzani, V. D. S. 5; Castro, N.

G. 1
1
Laboratrio de Farmacologia Molecular , ICB/UFRJ
2
3
LASSBio - Faculdade de Farmcia, UFRJ
4
LASSBio - Faculdade de Farmcia, UFRJ
5
Ncleo de Bioensaios, Biossntese e Ecofisiologia (NuBBE), UNESP
6
Objectives:
LASSBio-767 is an acetylcholinesterase inhibitor with central action, composed by a mixture of (-)-3-O-acetyl-spectaline and (-)3-O-acetyl-cassine. Unlike the drugs used for the symptomatic treatment of Alzheimer's disease, it displays few peripheral
muscarinic adverse effects, suggesting additional mechanisms of action (Eur. J. Pharmacol. 580; 339, 2008). We have
investigated direct interactions with muscarinic M3 receptors in the HT-29 epithelial cell line from human colon, a model system
of gastrointestinal effects. Because the hippocampus has also muscarinic receptors, especially M2 and M1/3, which modulate
cognitive functions, we studied the effects of LASSBio-767 in central inhibitory synapses through electrophysiological
recordings in cultured rat hippocampal neurons. Additionally, to clarify the activity of this substance on the M2 muscarinic
receptors, experiments were carried out in the isolated rat heart, based on the method of Langendorff, to verify the effect of
LASSBio-767 on heart rate.
The action of LASSBio-767 and its isolated constituent (-)-3-O-acetyl-cassine was tested by calcium fluorimetry in HT-29 cells
expressing M3 muscarinic receptors. Both inhibited the calcium response induced by carbachol, with similar Kbs of 0.69 M
and 0.50 M, respectively. The recordings of GABAergic tonic current (GABAtc) in neurons of rat hippocampus were performed
using the patch-clamp technique in whole-cell mode, with membrane potential set at 80 mV. We used different protocols in
which it was possible to observe the presence of GABAtc, which was blocked by 50 M bicuculline, a GABAa receptor
antagonist. LASSBio-767 30 M decreased GABAtc by 42.0 33.6 pA (mean SD, N=7) in comparison with the control
solution. In the presence of bicuculline it also decreased the current by 32.3 21.4 pA (N=3). In the presence of carbachol,
LASSBio-767 decreased total tonic current by 14.8 16.8 pA (N=7), and it still slightly decreased the total current in the
combined presence of carbachol and atropine, by 6.5 14.6 pA (N=7). Therefore, there was no evidence of a presynaptic effect
of LASSBio-767 on tonic GABA release, but it may have inhibited inward currents in the recorded neurons. The contraction of
the isolated rat heart was measured by a tension transducer. The test was done with 300 M methamidophos, an
organophosphorus pseudo irreversible inhibitor of acetylcholinesterase, and 30 M LASSBio-767. Methamidophos perfusion
decreased the frequency during 10 minutes of recording, with average rate of change of 2.85 0.25 bpm/min (N=3). LASSBio767 also decreased the frequency at the same time of perfusion, with average rate of decay of 3.70 0.44 bpm/min (N=3).
Conclusions:
LASSBio-767 and (-)-3-O-acetyl-cassine competitively inhibited the M3 receptor-dependent calcium increase induced by
carbachol in an epithelial cell and this may explain the lack of muscarinic hypersecretory effects in vivo, typical of other
acetylcholinesterase inhibitors. The electrophysiological results did not show a clear effect of LASSBio-767 in M2 receptordependent tonic GABA release. The experiments with rat heart showed a possible muscarinic cholinomimetic effect. However,
whether this effect is due to acetylcholinesterase inhibition or activation of M2 receptors remains to be clarified.
Keywords: acetylcholinesterase inhibitor , Alzheimer, GABAergic tonic current , muscarinic receptors
Financial Support: CNPq, FAPERJ and Finep.
Resumo:29-072
SOLUBLE NTPDASE ACTIVITY IN A MOUSE MODEL OF CONTACT DERMATITIS
da Silva, G. L. 2; Sperotto, N. D. M. 2; Battastini, A. M. O. 3; Campos, M. M. 2; Zanin, R. F. 2; Morrone, F.

B. 2
2
Departamento de Farmacologia Aplicada, PUCRS
3
Objectives:
Aim: Croton oil is a chemical irritant that causes swelling when applied topically. Nucleotides released by chemically injured
keratinocytes may function as pro-inflammatory mediators in skin, by acting on keratinocytes and other cell types (Mizumoto et
al., Nat. Med., 8; 358-365, 2002). The croton oil-induced mouse ear edema is a well established contact dermatitis model. The
aim of this study was to investigate the effects of A438079 (a P2X7 antagonist) and apyrase treatment on ATP, ADP and AMP
hydrolysis in mice serum, following topical ear application of croton oil.
Methods and results: Swiss mice (n=5) received topical application of 1% croton oil on the right ear, and vehicle on the left ear.
The animals were treated with A438079 i.p. (80 mol/kg) or apyrase s.c. into the ear (0.2 U/ear), 30 min before and 3 hours after
croton oil painting. Dexamethasone s.c. (0.5 mg/kg) was used as positive control. Six hours after the edema induction, blood
samples were collected and the animals were examined for ear swelling responses. Blood samples were collected and
immediately centrifuged at 3,000 g for 10 min at room temperature and the serum samples were used to measurement of
nucleotides hydrolysis. Apyrase treatment showed an edema inhibition of 17.6% 4.5, whereas A438079 decreased the edema in
41.7% 5.3. Croton oil decreased ATP and ADP hydrolysis in serum, and these effects were reversed by all treatments. AMP
hydrolysis was not altered by croton oil application, or even by apyrase and A438079 treatments, but it was significantly
increased by dexamethasone treatment, as previously described.
Conclusions:
Conclusion: The control of nucleotide and nucleoside levels exerted by ectonucleotidases could contribute to the modulation of
purinergic signaling, promoted by purinergic receptors, during inflammatory events. Our results showed a decrease in the ATP
and ADP hydrolysis following topical inflammation induced by croton oil, suggesting that the enzymes involved in this process
might act in the regulation of extracellular nucleoside/nucleotides, during skin inflammation. Furthermore, we demonstrated a
nucleotide-mediated pathogenic mechanism and a probable involvement of P2X7 receptor in the contact dermatitis induced by
croton oil.
Keywords: Purinergic receptors, P2X7, Apyrase, Inflammation, Dermatitis
Financial Support: Capes, CNPQ
Resumo:29-073
ALCOHOL USE MOBILIZES EXTRACELLULAR CA2+ IN VAS DEFERENS OF YOUNG MALE RATS.
Verde, L. F. ; Lopes, G. S. ; Jurkiewicz, N. H. ; Caricati-neto,a. ; Jurkiewicz, A.

UNIVERSIDADE FEDERAL DE SO PAULO, UNIFESP
Objectives:
In previous studies we showed that alcohol use during pregnancy and nursery causes a decreased of cytosolyc calcium and
contractility under the action of KCl and NE in rat vas deferens (RVD) of descendent male litters (SBFTE;64,2009). Now, our
objective was to check if acute treatment of rats with alcohol causes alteration in translocation of calcium and of tension induced
by KCl and NE in RVD of 40-days old litters.
Wistar rats, 40-days old litters, were treated with single dose of alcohol (i.p. 3.0 g/Kg) 4 hours before sacrifice. Controls received
intraperitoneal saline. Longitudinal strips of prostatic portion of the RVD of litters were mounted in PTI system (USA) and
intracellular calcium and contractions were evaluated by simultaneous measurements of fluorescence and tension in strips loaded
with fura-2. The mean values ( EPM) of simultaneous measurements of fluorescence ratios (R340/380(%) and tension (T%)
evoked by KCl (80mM) were significantly lower in treated (40,3 8,5 and 0,6 0,09 (5)) than in control (118,0 32,7 and 1,3
0,3 (4)), and for NE ( 50,2 4,15 and 0,48 0,13 (4)) and (118,5 30,7 and 0,98 0,15 (2)) in treated and controls respec tively.
Conclusions:
The present results suggest that alcohol use induced an alteration in translocation of calcium in RVD of males, possibly affecting
the mobilization of calcium intra and extracellular.
Keywords: alcohol, calcium, vas deferens, rats
Financial Support: Fapesp, CNPq and Capes
Resumo:29-074
SUCCINATE DEHYDROGENASE: MODULATION BY ADENOSINE NUCLEOTIDES IN SOLANUM TUBEROSUM
Jardim-messeder, D. ; Camacho-pereira, J. ; Galina, A.

IBqM/ Universidade Federal do Rio de Janeiro, UFRJ
Objectives:
Potato tuber mitochondria (PTM) have a succinate dehydrogenase (SDH) activity in matrix face of mitochondrial inner
membrane. This enzyme can be modulated by the adenylate nucleotides balance, but the mechanism of this regulation is not
completely understood . The enzyme adenylate kinase (ADK), can be able to regulate the AMP/ADP/ATP balance, and can
participate of the SDH modulation. The objective of this work is to investigate the mechanism of this regulation.
Using DCIP assay, it was verified that ADP and AMP increase the SDH activity in 50% and 70%, respectively, and this
stimulatory effect is higher in low concentrations of these nucleotides (50 M-100 M). However, in presence of Ap5A, a
specific inhibitor of ADK, the ADP activation was prevented, indicating that the stimulatory effect is mainly due to AMP. The
incubation with ATP is not able to increase the SDH activity. In addition, succinate dependent oxygen consumption in presence
of atractyloside (ATR), an inhibitor of the adenosine nucleotide transporter (ANT), is inhibited by ATP and ADP in about 50%,
but not by AMP. These data suggest that the inhibitory effect promoted by nucleotides is dependent on the inner mitochondrial
membrane side and is not related to membrane potential. ATP also inhibits SDH oxidation activity when we measured FAD+
red/ox levels by redox fluorimetric analyses. The inhibition of SDH is accompanied by an increase in reactive oxygen species
(ROS) production with ADP e ATP. ADP and ATP also inhibit the external NADH dehydrogenase from PTM but promote no
effect in mitochondrial complex I activity.
Conclusions:
In mitochondrial matrix, AMPand ADP are able to increase the SDH activity. However the stimulatory effect of ADP is
dependent of AMP production by ADK. However in cytosolic face of the inner mitochondrial membrane, ADP and ATP inhibit
the succinate dependent oxygen consumption.
Keywords: Adenoside Nucleotides, Potato Tuber Mitochondria, Succinate Dehydrogenase
Resumo:29-075
ANTAGONIC EFFECT OF AMPHETAMINE VERSUS ETHANOL ON ADRENERGIC NEUROTRANSMISSION IN
THE YOUNG RAT VAS DEFERENS
Silva Jnior, E. D. ; Caricati-neto, A. ; Jurkiewicz, N. H. ; Jurkiewicz, A.

Departamento de Farmacologia, Unifesp/EPM
Objectives:
Because of the few studies that emphasize the in vivo acute treatment with amphetamine and ethanol, and their consequence on
noradrenergic transmission in the smooth muscles of young animals, we decided to study the effect of these drugs on peripheral
sympathetic neurotransmission. We used the vas deferens (VD) of young rats as a model for the study of sympathetic
neurotransmission.
The young animals were divided into two groups. Amphetamine (AMPH) was administered at doses of 3 mg.kg-1, while ethanol
(EtOH) was administered at doses of 1.2 mg.kg-1. Amphetamine and ethanol were administered either alone or simultaneously
(AMPH 3 mg.kg-1 + EtOH 1.2 mg.kg-1) to investigate possible interactions. For functional experiments, the VD was mounted
under 1 g tension in isolated organ bath. The contraction was recorded in physiograph. Cumulative concentration-effect curves
were made for adrenergic agonists (noradrenaline, dopamine, phenylephrine) and barium. Pharmacological parameters Emax,
pD2 and were analyzed. Time-response curves were also performed for tyramine, to check the release of endogenous
noradrenaline. To evaluate the neurogenic contraction to electrical field stimulation (EFS) the VD was mounted on the isolated
organ bath between two electrodes. Stimuli of 60 V, 1 ms duration at frequencies of 0.1 to 20 Hz were employed. To evaluate the
role of calcium the VD was depolarized for 5 minutes with 80 mM KCl in the absence of Ca+2 in the presence of EGTA (10M)
followed by the addition of a single dose of CaCl2 (10 mM for 10 minutes). For the dosage of noradrenaline we used an
electrochemical detector for the detection of catecholamine in HPLC, Shimadzu, column RP-18e Chromolith (Merck). Results:
The group treated with AMPH 3 mg.kg-1 showed a potentiation of the VD contractile response of noradrenaline, barium and
calcium, without change of neurogenic contraction. It was noted also that the content of noradrenaline was reduced. The group
treated with EtOH 1.2 g.Kg-1 showed a decrease in VD contractility to noradrenaline, phenylephrine and barium, and EFS, as
well as a lower content of catecholamine. The group treated with AMPH 3 mg.kg-1 + EtOH 1.2 g.kg-1 did not show any changes
in experimental protocols used. These data suggest that amphetamine and ethanol could be acting antagonistically. The treatments
with amphetamine or ethanol and simultaneous treatment did not significantly increase plasma concentrations of corticosterone.
Conclusions:
We observed a possible antagonism between amphetamine and ethanol when administered simultaneously on peripheral
sympathetic neurotransmission of young animals.
Keywords: Amphetamine, Antagonic effect, Ethanol, Neurotransmission, young rat
Financial Support: CAPES - Coordenao de Aperfeioamento de Pessoal de Nvel Superior
Resumo:29-076
INTRACEREBROVENTRICULAR LPS DEPHOSPHORYLATED HYPOTHALAMIC AMPK BUT IT DID NOT
ALTER HEPATIC PEPCK EXPRESSION
Vitorino, D. C. 1; Roman, E. A. 1; Santos, G. A. 2; Pereira, V. D. 2; Moura, R. F. 2; Velloso, L. A. 3;

Milanski, M. 4; Torsoni, M. A. 4
1
Physiology / IB, Unicamp
2
Medical Clinical / FCM, Unicamp
3
Internal Medicine / FCM, Unicamp
4
Central of health and biochemistry / FCA, Unicamp
Objectives:
AMP-activated protein kinase (AMPK) has long been identified as a critical regulator of energy homeostasis in metabolic tissues.
The high consumption of saturated fat as well as the presence of pathogenic organisms can lead to an increase energy expenditure
and satiety through toll like receptor-4 activation and inflammatory cytokine expression. Thus, AMPK modulation following the
TLR-4 activation could be an important event related to several metabolic diseases as obesity and type 2 diabetes. Therefore, the
aim of this study is to evaluate the TLR-4 activation effects on AMPK hypothalamic pathway and glucose homeostasis.
Methods: Wistar rats and Swiss mice were used. Rats were submitted to stereotaxic surgery to implant of guide cannula into the
lateral ventricle. Lypopolissacaride was administered via intraperitoneal (IP) or intracerebroventricular (ICV) (75 g/animal-ICV
or 1 mg/Kg-IP) and later the rats were killed by decapitation and the hypothalamus was quickly removed and stored at -80C.
When necessary the food intake was measured during dark period. The protein analysis was performed by western blot and blood
glucose was determined by glucose oxidase method. Results: The food intake in animals treated with LPS (via ICV or IP) was
significantly reduced compared to control animals (ICV:50% and IP: 60%). However, blood glucose in LPS animal compared to
control group was reduced after IP treatment, but did not change after ICV treatment. Besides, liver PEPCK expression was not
modified by ICV injection but it was reduced by IP treatment. The AMPK phosphorylation was significantly reduced by both
treatments (ICV and IP) compared to control group.
Conclusions:
Hypothalamic AMPK play important role in the satiety signal and energy expenditure, two events that can be damaged model of
sepsis and obesity. The resulted showed that although LPS treatment (ICV) inhibited the AMPK activation and reduced food
intake in fasted rats, glucose homeostasis (blood glucose and liver PEPCK expression) was not modified, suggesting that direct
effect of LPS on the liver is necessary to inhibit glucose production.
Keywords: AMP-activated protein kinase, hypothalamus, inflammation, Lypopolissacaride, TLR-4
Resumo:29-077
EFFECTS OF 17-ESTRADIOL, ICI 182,780 AND G1 ON ERS1-NEGATIVE CANCER CELLS (SKBR3)
Sousa, A. 2; Luna, M. S. A. 2; Porto, C. S. 3; Yamanouye, N. 2

2
Lab. Farmacologia, Instituto Butantan, IBU
3
Depto. Farmacologia, UNIFESP, UNIFESP
Objectives:
G protein-coupled estrogen receptor 1 (GPER, or GPR30), which belongs to the family of seven-transmembrane G proteincoupled receptors, mediates the rapid (non-genomic) signaling of 17&beta-estradiol and is involved on proliferative effects of
17&beta-estradiol in breast cancer cells. The aim of this study is to analyse the role of GPER in breast cancer cells.
An ERS1-negative cancer cells (SKBr3) that express GPER was used in this study. These cells were cultured in RPMI medium
with antibiotics and 10 % FBS. 24 hours before the experiments, the cells were maintained in RPMI medium without FBS and
phenol. Afterwards, the cells were stimulated with 17-estradiol (1nM), ICI 182,780 (1nM - antagonist of ERS1 and ERS2, but
agonist of GPER) or G1 (1 nM - specific agonist of GPER) and the signaling pathway and protein expression were evaluated. The
effect of 17&beta-estradiol on MAP kinase (ERK 1/2) or Akt was performed by Western blotting, using an antibody specific to
the phosphorylated (activated) form of this protein. 17&beta-estradiol was able to activate ERK1/2 and the maximum activation
occurred after 5 minutes of incubation, but not Akt. Protein expression was also evaluated using bidimensional eletrophoresis (2DE) and the images were analyzed by Image Master Platinum 7 software. The 2-DE images of cells extracts showed stained spots
with PI ranging from 3 to 10 and molecular mass ranging from 216 to 7 kDa in all treatments and in the control. 17&betaestradiol, ICI 182,780 or G1 (2h, 37oC) were able to promote differential protein expression in cells SKBr3, when compared to
control cells (211, 250, 318 and 44 specifics spots were found in 17&beta-estradiol, ICI 182,780 or G1 treated cells or control
cells, respectively). When the protein expression was compared among the treatments, specifics spots were observed for each
treatment (17&beta-estradiol 191, ICI 182,780 415, G1 354) and 18 spots were expressed in all treatments. Between
17&beta-estradiol and ICI 182,780 or 17&beta-estradiol and G1, 38 and 251 spots were expressed in both treatments,
respectively. However, no common spots were observed between ICI 182,780 and G1 treatments.
Conclusions:
These results showed that stimulation of GPER activated rapid signaling and protein expression in SKBr3 cells. In addition, these
results also suggested that in SKBr3 cells have other estrogen receptors besides GPER, since different treatments promoted
different protein expression.
Keywords: estrogen receptor, proteomic analysis, SKBr3 cells
Resumo:30-021
"IN VITRO ANTITUMORAL ACTIVITY OF A LIPID NANOEMULSION CONTAINING A CURCUMIN
DERIVATIVE"
Romeiro, J. G. 1,2; Santos, M. A. 1,3; Valduga, C. J. 1,3; Diniz, S. N. 1,2

1
Dep. de Farmcia da Universidade Bandeirante de So Paulo/SP, UNIBAN, So Paulo
2
Laboratrio de Biologia Celular e Molecular, LBCM
3
Laboratrio de Sntese e Formulaes, LSF
Objectives:
Aim: Natural agents with potential anticancer activity include curcumin, a natural compound extracted from turmeric (Curcuma
longa). The 1,5-bis(4-hydroxy-3-methoxyphenyl)-penta-1,4-dien-3-one (1) is a phenolic compound with similar curcumin
structure, that showed in vivo low toxicity and significant antitumoral activity. The use of various antitumor compounds is
restricted because of its inespecific cytotoxic activity. In the last decades, great investments have been made in the development
of drug delivery systems for the treatment of diverse diseases including cancer. Many works has been proved the use of lipid
based nanoparticles to carry antineoplastic agents with low water solubility. Here we developed an emulsion formulation for a
lipophilic diacyl derivative (1,5-bis(4-oleyl-3-methoxyphenyl)-penta-1,4-dien-3-one, 2) of compound 1 and evaluated its physicochemical properties, stability and in vitro cytotoxicity against leukemic cell tumor lineage.
Methods and Results: Two oleyl groups were attached in each phenolic hydroxyl group of compound 1 through an esterification
reaction in the presence of dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP) as catalyst, obtaining 100 %
of yield. The nanoemulsion composition was phosphatydylcholine 44.9 %, triglyceride (cotton-seed oil) 22.5 %, cholesterol 0.6
%, Tween 20 18.0 %, b-carotene 0.6 % and compound 2 13.4 %. The mixture was emulsified under 10,000 psi of pressure at 50
oC for 20 min. The incorporation yield of compound 2 was 100 %, the particle size average was 85 nm with approximately 0.2 of
polydispersity. Both formulations (with or without compound 2) maintained their stability for six months; without aggregation,
precipitation or color change. During this period it was not observed lipid peroxidation, but pH changed from 5.3 to 4.47,
indicating a slight hydrolysis of the triglycerides. In kinetic experiments through dialysis of the compound 2 against plasma in
vitro, we observed a dissociation ratio of 30% in the first hour of experimentation. The cytotoxicity studies were performed on
leukemic cell line Jurkat using "MTT" (3-4,5-dimethylthiazol-2,5-diphenyltetrazolium) method in three independent experiments.
The results of the nanoemulsion containing compound 2 demonstrated a significant cytotoxic activity (p = 0.025, unpaired t test)
compared with the nanoemulsion without the active agent. Future experiments should be done in normal T cells to evaluate the
specificity of the nanoemulsion and in other tumor cell lines to confirm its antitumor activity.
Conclusions:
Conclusions: In this study we developed a lipid based nanoemulsion containing a lipophilic derivative of compound 1, a
curcumin analogue, which showed significant antitumor activity in leukemic cells.
Keywords: curcumin derivative, lipid nanoemulsion, leukemic cells, antitumoral activity
Financial Support: UNIBAN / FAPESP
Resumo:30-022
CHARACTERIZATION AND COMPARISON OF THE CELL CYCLE CONTROL OF THREE DIFFERENT
EMBRYONIC STEM CELL LINEAGES
Assis, J. L. 2; Fragel-madeira, L. 1; Mariante, R. M. 2; Rehen, S. K. 3; Linden, R. 2

1
Departamento de Neurobiologia Instituto de Biologia - UFF, UFF
2
Instituto de Biofsica Carlos Chagas Filho - UFRJ, UFRJ
3
Laboratrio Nacional de Clulas-Tronco Embrionrias, LANCE - UFRJ
Objectives:
Stem-cells have unlimited or prolonged capacity to self-renewal in an undifferentiated state, and are able to produce more than
one kind of highly differentiated descendents. Although having a normal diploid karyotype and low mutation frequency their
growth is similar to that of malignant cells, whose proliferation is not sensible to serum deprivation nor is inhibited by contact or
subject of anchorage. Despite of the large amount of attention that has been given to embryonic stem cells and their potential uses
in cell therapy, we know little about the singular behavior of these cells. Thus, sparing knowledge leads to lack of safety for the
use of these cells in medical therapy. Identification of proteins that are involved in controlling proliferation and regulation of the
cell cycle in embryonic stem cells is essential to understanding the mechanisms for regulating the growth, cellular transformation
and maintenance of the pluripotency. Cyclins are short lived proteins involved in the control of cell cycle progression and form
functional complexes with the cyclin-dependent-kinases (CDKs). The INK4-family and the Cip1/Waf1/Kip1-2-family are
inhibitors sensitive to several signals, for instance DNA damage, and are able to inactivate these complexes, thus activating the
checkpoint machinery and preventing cell proliferation.
Therefore, with the goal of exploring the cell cycle control of murine embryonic stem cells (mES), we analyzed the presence and
localization of regulatory proliferation proteins, besides the cell cycle distribution in pluripotent mES, comparing three different
lineages: USP1, R1 and miPS, exposed or not to mutagenic agents, hydroxyurea (HU) or ultraviolet C (UV-C) irradiation (20
J/m2). Moreover, we did the same analyses with embryonic bodies (EBs) of these three mES lineages committed to the neural
phenotype, by treatment with retinoic acid. Then the cells were processed to immunocytochemistry and viewed by fluorescence
microscopy or submitted to flow citometry analysis. Our results showed that the mES remained undifferentiated on long-term
culture, confirmed with immunocytochemistry by presence of pluripotency markers OCT-4 and SSEA-1. These undifferentiated
cells, not synchronized for any specific phase cell cycle, expressed cyclins A, B1, D1, E, CDK2 and CDK6, confirmed by
immunocytochemistry. In addition to these, we also found CDK4, p27 and p16 absent. Interestingly, cyclins D1, E, A and CDK6
had presented nuclear localization but cyclin B1 was located in the cytoplasm. The content of cyclins A and B1 were found
increased in USP1 after HU treatment by flow citometry analysis. In addition, both HU and UV-C treatments induced an arrest in
the USP1 cell cycle, confirmed by propidium iodide (PI).
Conclusions:
These results suggest that the presence and localization of such cell cycle regulators must have an important role for the
maintenance of mES in a proliferative status and both treatments with mutagenic agents prejudice the undifferentiated phenotype.
Keywords: Embryonic Stem Cell, Cell Cycle Control, Cyclins
Financial Support: CNPq, MCT/DECIT, PRONEX, FAPERJ.
Resumo:30-023
EFFECTS OF THE PLATELETACTIVATING FACTOR ON THE PLURIPOTENCY OF MURINE EMBRIONIC
STEM CELLS
Signoretti, P. V. P. ; Fragel-madeira, L.
Neurobiologia/Universidade Federal Fluminense, UFF
Objectives:
Cell therapy to treat neurological lesions with the use of embryonic stem cells present great potential to the functional repair. As
described, the intracellular signal pathways are essential to the growth of mES in the presence of leukemia inhibiting factor (LIF),
through the activation of the LIF receptor (LIFR), to the conservation of the pluripotency and self-renewal of these cells. Because
of its pluripotentiality, however, these cells can behave as malignant cells and become tumors after the tissue transplantation. The
platelet-activating factor (PAF) is a phospholipid that shows several biological activities, including proliferation and cell
differentiation. PAF signaling occurs through a receptor coupled to the G protein (PAFR) that exists in the plasma membrane.
However its effect on the cell proliferation is somewhat controversial, depending on the cell type and distinct signaling pathways.
Our main goal with this proposal is to demonstrate the effect of PAF on the cell cycle of murine embryonic stem cells (mES)
through analysis of signaling pathways resulting from activation of PAFR in presence and absence of LIF. Until now there are no
reports upon the effects of PAF on the mES or the description of the presence of its receptor in these cells.
Murine embryonic stem cells (USP1 line) were co-culture with mouse embryonic fibroblasts (MEF) inactivated with Mitomycin
C and analyzed by immunocytochemistry for pluripotency markers OCT-4 and SSEA-1. The presence of PAFR in mES and MEF
was analyzed by western blot. mES was treated with PAF at various concentrations and analyzed for cellular viability and death
through propidium iodide and proliferation assay by BRDU incorporation. Our preliminary results showed that the mES
remained undifferentiated in long-term culture confirmed by immunofluorescence for OCT-4 and SSEA-1 and absence of the
neural progenitor marker Nestin. Beyond that mES culture over MEFs expressed PAFR as analyzed by western blotting. We
found that the treatment with some concentrations of PAF decreased the number of cells in absence of LIF. We also noted in
absence of LIF that the percentage of OCT-4 positive cells was the same after treatment with PAF, but there were less cells
incorporated BrdU compared to control, suggesting a decrease in the proliferation rate instead of cell death. In contrast, in the
presence of LIF, the OCT-4 positive cells increased BrdU incorporation after PAF treatment although did not change the
percentage of OCT-4 positive cells.
Conclusions:
Based on our results PAF treatment in combination with LIF seems to maintain the pluripotent state of mES and control their
proliferation. Thus, it is necessary to study the signaling pathways triggered by both factors in understanding the pluripotency of
stem cells due to their tumorigenic potential.
Keywords: LIF, MURINE EMBRIONIC STEM CELLS, PAF, PLURIPOTENCY
Financial Support: Faperj, Capes, Proppi-UFF
Resumo:30-024
BONE MARROW MONONUCLEAR CELL THERAPY REDUCE HIPPOCAMPAL CA1 PYRAMIDAL CELL
DEATH FOLLOWING GLOBAL CEREBRAL ISCHEMIA IN THE RAT.
Ramos, A. B. 1; de Vasconcelos, A. 1; Cintra, W. M. 2; Mendez-otero, R. 1

1
Laboratrio de Neurobiologia Celular e Molecular, IBCCF / UFRJ
2
Laboratrio de Famarcologia, ICB / UFRJ
Objectives:
The pyramidal neurons of the hippocampal CA1 (Cornu Ammonis) region are essential for cognitive functions and are selectively
destroyed after global cerebral ischemia. The aim of our study was to investigate whether bone marrow mononuclear cells
(BMMCs) treatment could protect against neuronal damage observed after transient global ischemia.
Wistar male rats of 8 week-old, weighing 250-300g were submitted to transient forebrain ischemia by the four vessels occlusion
(4VO) method. In order to establish the time course of neuronal death in CA1 region, we used a marker of early neuronal
degeneration, Fluoro-Jade C (FJ-C). Rats were anaesthetized, transcardially perfused with 4% paraformaldehyde and their brains
were dissected 3, 7 or 14 days after ischemia (DAI). Coronal slices of the hippocampus were made (3.8mm 4.3mm bregma
approximately) for the histological processing. We observed a greater number of cells labeled with FJ-C in the CA1 region of
hippocampus of animals analyzed 7 DAI (159.1 22.77 cells/ mm CA1) when compared with the animals analyzed 3 or 14 DAI
(14.12 0.89 cells/mm CA1; 114.3 16.50 cells/mm CA1, respectively). BMMCs obtained from femurs and tibias of adult rats
(3x10^7 cells in 300 l saline) or saline only (300 l) were injected into the left carotid artery 1 or 3 DAI and the animals were
then analized 7 DAI to FJ-C histochemistry. We observed a significant reduction of cell death in CA1 region of BMMCs treated
animals, in both 1 and 3 DAI therapeutic windows (66.67 10.26 cells/mm CA1; 78.33 16.63 cells/mm CA1, respectively)
when compared with the group that received saline (177.6 69.92 cells/mm CA1). In some experiments BMMCs were incubated
with Cell Trace, a fluorescent tracer for very long-term cell labeling. Cell Trace positive cells were observed near the CA1 region
of animals treated with BMMCs.
Conclusions:
We conclude that intra-arterial administration of BMMCs after transient global ischemia reduces hippocampal damage.
Keywords: global ischemia, stem cell , hippocampus
Financial Support: Coordenao de Aperfeioamento de Pessoal de Nvel Superior - CAPES
Resumo:30-025
CITOTOXICITY OF CHLORINEE6 INCORPORETED IN A NANOEMULSION LIPIDIC IN HUMAN MELANOMA
CELLS LB373-MEL
Macaroff, P. P. 1; Greene, L. J. 1; Tedesco, A. C. 2

Faculdade de Medicina de Ribeiro Preto-USP, FMRP-USP
2
Faculdade de Filosofia Cincias e Letras de Ribeiro Preto, FFCLRP-USP
1
Objectives:
Photodynamic therapy (PDT) is a minimally invasive therapeutic approach approved for the treatment of neoplastic and vascular
diseases. It consists of (i) a photosensitizer that can be applied topically or administered systemically; (ii) visible light, usually
generated by laser a source and (iii) molecular oxygen, which is used in the photodynamic reaction generating singlet oxygen
(1O2) and reactive oxygen species. The objective of this study was to evaluate citotoxicity of photosensitizer Chlorine e6
incorporated in a nanoemulsion lipidic (LDE-Ce6) on a culture of human melanoma cells LB373-MEL after irradiation with a
light laser.
The in vitro assays were conducted in the dark and in the presence of light laser at 650 nm adjusted from 0.5 to 10 J/cm2 at 300
mW power. Citotoxicity, LDE-Ce6, was determined with the MTT assay. Statistical analysis was performed using Prism 5 by
one-way ANOVA, and Newman-Keuls test as the mean SEM of three experiments (p < 0.05). Previously studies indicated that
5 M Chlorine e6 for 2 hours incubation provided maximal up-take. No cytotoxicity was detected in cells in the absence of the
light laser. Cell damage was dose dependent on the energy of light: 50% cell death at 0,5 J/cm2, 70% cell death at 2 J/cm2, 85%
cell death at 5 J/cm2 and 97% cell death at 10 J/cm2.
Conclusions:
The present study shows that the cell line LB373-MEL is susceptible to PDT action using a Chlorine e6 5M incorporated in a
nanostructured drug delivery system for 2 hours incubation and application light laser at 10J/cm2. In addition, the conditions
identified here will be used with proteomic analysis of cell line LB373-MEL treated by PDT.
Keywords: Melanoma, Photodynamic therapy , Proteomic analysis
Financial Support: FAPESP, FINEP and CNPq
Resumo:30-026
ANTIFUNGIC ACTIVITY OF DIRECT ELECTRIC CURRENT ON CANDIDA ALBICANS
Barbosa, G. M. 2; Santos, E. G. 2; Maral, C. P. 2; Valle, R. S. 2; Abi-chacra, E. A. 2; Veiga, V. F. 2; Santos,

A. L. S. 2; Holandino, C. 2
3
Instituto de Microbiologia Professor Paulo de Ges, UFRJ
2
Departamento de Medicamentos / Faculdade de Farmcia, UFRJ
Objectives:
The aim of this work was to evaluate the effect of electrotherapy on Candida albicans, since cell death promoted by direct
electric current (DC) has been reported in many cases, such as antitumoral and bactericidal effects.
Yeasts (106 cells/mL) were submitted to cathodic flow (CF), electroionic flow (EIF) and anodic flow (AF) for 1 to 20 min,
varying the DC from 0.5 to 2.0 mA. After these procedures, the cellular viability was measured by incorporation of propidium
iodide (PI) or by plating cells on nutrient agar to measure the colony-forming units (CFU). After 5 min of DC at 0.5, 1.0 and 1.5
mA, we did not detect any CFU in AF in three independent experiments. The same occurred after 1 min of DC at 2.0 mA.
However, only a reduction by 50% in cellular viability with PI was observed after 5 min at 0.5 mA. Comparing with control, CF
and EIF presented similar values of CFU under the experimental conditions assayed. Biochemical alterations related to the
expression of secreted aspartic peptidases (Saps) by Candida albicans were evaluated after exposure of yeasts to DC. Sap1-3
expression was evaluated in cell surface by flow cytometry. Additionally, the aspartic protease activity was measured using BSA
as protein substrate. After 20 min of DC at 2.0 mA, we observed a reduction of 70% and 30% in the expression of surface Sap1-3
after exposition of yeasts to CF and EIF, respectively, in comparison to the untreated cells. This result was corroborated through
qualitative analysis of the protease activity. New experiments are underway to understand the antifungal activity of DC, including
transmission electron microscopy in order to evaluate the possible morphological alterations.
Conclusions:
DC decreases the cellular viability of C. albicans, presenting high susceptibility to AF. Furthermore, DC is able to modify other
metabolic mechanisms of yeasts in CF and EIF, such as protein expression directly related to infection process, although cell
viability is not altered. These results demonstrate the potential of electrotherapy to treat cutaneous and subcutaneous fungal
infections.
Keywords: Candida albicans, Cellular viability, Direct electric current, SAPs
Resumo:30-027
SENSORY-MOTOR RECOVERY INDUCED BY THE TREATMENT WITH BONE MARROW MONONUCLEAR
CELLS AFTER UNILATERAL FOCAL ABLATION OF THE CEREBRAL CORTEX IN RATS.
Gomes da Silva,v. ; de Freitas,h. T ; Giraldi-guimares A.

Laboratrio de Biologia Celular e Tecidual, UENF
Objectives:
Aim: Focal lesions of the cerebral cortex induced by different protocols lead to different effects in functional loss and plasticity
of connections. Focal cortical lesion induced by ischemia promotes plasticity of cortical connections. However, focal cortical
lesion induced by ablation does not, albeit with the same extension. Furthermore, ablation and ischemia induce different
functional impairments. It has been shown that therapy with bone marrow mononuclear cells (BMMCs) results in functional
recovery after focal cortical ischemia. The objective of this work was to investigate whether treatment with BMMCs promotes
the same functional recovery in a model of focal cortical lesion by ablation.
Methods: We used male Wistar 2-3 months (240-380g). Brain injury was induced by the removal by suction of most of the
primary motor cortex and part of the primary somesthetic cortex of the left hemisphere. The animals were splitted in two groups:
control (n=12) and treated with BMMCs (n=10), and approx. 24 hours after the induction of the lesion, they were treated with
PBS and approx. 3 x 107 mononuclear cells extracted from bone marrow of donor rats, respectively. The administration was by
injection through the jugular vein. Two sensorimotor tests were used to estimate functional recovery and were performed before
and after injury followed by injection of cells or PBS. In cylinder test, each animal was placed inside a glass cylinder and an
asymmetry rate of the use of the forelimbs during vertical exploration was calculated. In the adhesive test, pieces of sticky paper
were placed bilaterally to the bottom of the forepaws and the bias in the first removal was quantified. Data were statistically
evaluated by analysis of repeated measures. Results: The cylinder test showed significant effect of time and treatment, with
significant difference between both experimental groups from the middle of the second month after ablation. The group treated
with BMMSs presented significant decrease in the asymmetry rate (approx. 46% lower at the post-ablation day 91 in relation to
control group), which means a tendency of return to the initial pre-ischemic stage (asymmetry rate around 0 = symmetric use of
the forelimbs). In the adhesive test, similar result was observed. The percentage of first removal of the adhesive with the impaired
(right) forelimb was significantly higher in the group treated with BMMCs from the middle of the second month after ablation, in
relation to the control group.
Conclusions:
Conclusion: Our results suggest that the treatment with BMMCs is effective to induce recovery of sensorimotor function after
focal cortical lesion by ablation. Thus, BMMCs have therapeutic effect in a model of brain injury that does not induce significant
plasticity of the cortical connections. Further studies are necessary to investigate whether the recovery promoted by BMMCs
might be correlated to a possible induction of plastic changes in the cortical connections.
Keywords: cellular therapy, bone marrow mononuclear cells, ablation
Financial Support: UENF, FAPERJ.
Resumo:30-028
EARLY AND LATE EFFECTS OF BONE MARROW-DERIVED MONONUCLEAR CELL THERAPY ON LUNG
AND DISTAL ORGANS IN EXPERIMENTAL SEPSIS
Ornellas, D. S. 1,2; Maron-gutierrez, T. 1,2; Ornellas, F. M. 2; Cruz, F. F. 1; Lucas, I. H. 1; Fujisaki, L. 1;

Capelozzi, V. L. 3; Pelosi, P. 4; Morales, M. M. 2; Rocco, P. R. M. 1
1
1Laboratory of pulmonary investigation LIP, UFRJ / IBCCF
2
Laboratory of Cellular and Molecular Physiology LFCM, UFRJ / IBCCF
3
Department of Pathology, USP
4
Department of Surgical Sciences and Integrated Diagnostics, University of Genoa
Objectives:
This study tested the hypothesis that the early beneficial effects of bone marrow-derived mononuclear cells (BMDMCs) observed
on lung and distal organs were preserved late in the course of sepsis.
C57BL/6 mice were randomly assigned into two groups. Sepsis was induced by cecal ligation and puncture (CLP) surgery, while
a Sham operated group was used as control. One-hour after surgery, Sham and CLP groups were further randomized into
subgroups receiving saline (SAL) or BMDMC (2x10E6) intravenously (iv). All animals from CLP-SAL group died within 48
hours after sepsis induction. BMDMC therapy in CLP group at days 1 and 7 led to: 1) improved survival at day 1 (60%) and day
7 (75%) (n=7, p<0.05).
Conclusions:
BMDMC therapy was effective at preserving or improving the early beneficial effects on inflammatory and remodeling
processes, contributing to endothelium and epithelium alveolar repair resulting in improvement of lung mechanics. This potential
efficacy of BMDMCs for the treatment of sepsis may be associated to paracrine mechanisms.
Keywords: Cell therapy, growth factors, cytokines, apoptosis, sepsis
Financial Support: PRONEX-FAPERJ, CNPq, FAPERJ, CAPES, INCT-INOFAR
Resumo:30-029
IMPACT OF THYMULIN GENE THERAPY DELIVERED BY NANOPARTICLES ON LUNG REMODELING IN A
MURINE MODEL OF CHRONIC ALLERGIC INFLAMMATION
Silva, A. L. 1; Martini, S. V. 1; Abreu, S. C. 1; Samary, C. S. 1; Capelozzi, V. L. 4; Goya, R. 3; Boylan, N. 2;

Hanes, J. 2; Morales, M. M. 1; Rocco, P. R. M. 1
1
Instituto de Biofsica Carlos Chagas Filho/UFRJ, UFRJ
2
Johns Hopkins University - USA, JHU
3
National University of La Plata, UNLP
4
Faculdade de Medicina da USP, USP
Objectives:
Asthma is a common disease worldwide. And so far there has been no therapy able to revert the ultrastructural changes related to
airway remodeling. These unsuccessful attempts may be due to difficulty in delivering the therapeutic drug directly to the injury.
The aim of the present study is to analyse the effects of biologically active analog gene of thymulin, methionine-FTS, delivered
by nanoparticles on airway and lung parenchyma remodeling and repair in a murine model of chronic inflammation.
42 BALB/c mice were randomly assigned to six groups. In the OVA group, mice were immunized with 10 g ovalbumin (OVA,
ip) on 7 alternate days, and after day 40 they were challenged with three intratracheal instillations of 20 g ovalbumin at 3-day
intervals. Control mice (C) received saline under the same protocol. Twenty-days before the first challenge, C and OVA groups
were treated with plasmid encoding thymulin analog gene compacted with a nanoparticle (cationic polymer: lysine amino acid
conjugated to polyethylene glycol) (100 g of DNA/50 l of saline) intratracheally (NSAL and NOVA). On the same day,
another group of mice received only plasmid, without nanoparticles(PSAL and POVA). 24 h after the last challenge, lung
mechanics, airway responsiveness, collagen and elastic fiber content in airway and alveolar septa, and histology (light and
electron microscopy) were analyzed. Lung static elastance (34%), airway hyperresponsiveness, total number of inflammatory
cells (59%), the amount of alveolar collapse (67%), collagen and elastic fiber content and the thickness of epithelial reticular
basement membrane were higher in OVA group compared to NOVA. No significant differences were observed between OVA
and POVA group.
Conclusions:
In the present model of chronic allergic inflammation, thymulin analog gene delivery by nanoparticles modulated the
inflammatory and remodeling processes improving lung mechanics. Conversely, the use of plasmid without nanoparticle resulted
in no respiratory effects. Therefore, the use of nanoparticles to deliver medicine to the lungs may be a promissing strategy for
asthma therapy, even though further studies are necessary.
Keywords: Asthma, Gene Therapy, Nanoparticles
Financial Support: CNPq, FAPERJ, PRONEX-FAPERJ
Resumo:30-030
COMPARISON ON THE WOUND HEALING EFFECTS OF NANOCRYSTALLINE SILVER DRESSING AND
PAPAIN GEL 2% IN RATS
Fioravanti-jr, G. A. 1,5; Ceolin, L. D. 2; Maia, L. D. A. 3; Morrone, F. B. 3,4; Corte, T. W. F. 3; Campos, M.

M. 1,4
1
PPG em Medicina e Cincias da Sade PUC RS, PPGMCS PUC RS
2
Hospital So Lucas da PUC RS, HSL PUC RS
3
Faculdade de Farmcia, FFARM PUC RS
4
Instituto de Toxicologia, INTOX
5
Saavedra Tecnologia em Sade, SAAVEDRA
Objectives:
Nanocrystalline silver have been demonstrated to have antimicrobial and anti-inflammatory properties, whereas papain is a
proteolytic enzyme, which has been used in different formulations for wound treatment in clinics (Burns, 36: 751, 2010). This
experimental study aimed at comparing the effects of nanocrystalline silver dressing and papain gel 2% in wound healing in rats.
Male Wistar rats (120 160g, 6 weeks) were used. All the experimental procedures were approved by the local Animal Ethics
Committee. Twenty-four rats were randomly divided into four groups (6 rats per group): Group 1, Saline; Group 2,
nanocrystalline silver (Acticoat, Smith&Nephew); Group 3, excipient Gel; Group 4, papain gel 2%. For the wound healing test,
the rats were anesthetized by a mixture of ketamine (100 mg/kg) and xylazine (10 mg/kg), by i.p. route. The dorsal hair was
shaved, and full-thickness wound of 1.5 x 1.5 cm2 was excised from the dorsum. In the experimental groups 1, 3 and 4, the
dressings were changed every day, whereas in the group 2, the dressing was changed every 3 days. All the animals were
evaluated on the 3rd, 6th, 9th, and 12nd days for wound-healing rating. Qualitative analysis revealed scab formation in the saline
group 1 in all evaluated periods, whereas notable bleeding was seen in the papain gel 2% group 3, until the 9th day. In the
nanocrystalline silver group 4, it was possible to observe a more contractive aspect of the wounds, when compared to the other
experimental groups. Of note, quantitative analysis of wound areas revealed a significant reduction of the scars, in the group
receiving nanocrystalline silver dressing. The percentages of inhibition in relation to the saline-treated group were: 38 6%; 31
10 %; 50 10 %; and 46 13 %, at 3, 6, 9, and 12 of evaluation. On the other hand, the papain gel 2% was able to significantly
reduce the wound area only at the 3rd of evaluation, when compared to the gel group, with an inhibition percentage of 27 8 %.
Conclusions:
In this work, based on both the qualitative and the quantitative analysis of wound areas, the nanocrystalline silver group showed a
better wound healing effect in view of a rapid contraction of wound, compared with the others groups, indicating a pro-healing
activity for nanocrystalline silver dressings. New experiments are being conducted in order to investigate the possible
mechanisms underlying the beneficial effects of nanocrystalline silver on cicatrization.
Keywords: wound healing, nanocrystalline silver, papain , rat
Financial Support: CNPq, CAPES and PROBOLSAS PUC-RS
Resumo:31-061
EFFECT OF AQUEOUS LEAVES EXTRACT OF SCUTIA BUXIFOLIA REISSEK ON NTPDASE AND ADENOSINE
DEAMINASE ACTIVITY IN LYMPHOCYTES FROM RATS
Gomes, J. L. 3; Dalenogare, D. 1; Pimentel, V. C. 1; Fiuza, T. L. 1; Schmatz, R. 1; Boligon, A. 2; Athayde, M.

2
; Moretto, M. B. 2; Morsch, V. M. 1; Schetinger, M. R. C. 1
1
Qumica/ Universidade Federal de Santa Maria, UFSM
2
Departamento de Anlises Clnicas e Toxicolgicas, UFSM
3
Universidade de Cruz Alta, UNICRUZ
Objectives:
The World Health Organization (WHO) estimates that 6580% of the population of the developing countries depends on
medicinal plants for basic pharmaceutical care. Scutia buxifolia Reissek belongs to the Rhamnaceae family and is popularly
known as coronilha. It is a native plant from South America, with a dispersion area that includes the Rio Grande do Sul state,
Brazil. Infusions of the root bark are popularly used as a cardiotonic, antihypertensive and diuretic. The nucleotides are
messengers that modulate the exocrine and endocrine systems, the vasculature and hemostatic mechanisms and musculoskeletal,
immune and inflammatory cells. Their levels are controlled by a complex cell surface-located group of enzymes called
ectonucleotidases, anchored in the plasmatic membrane of platelets. The cascade of ecto-enzymes, with this function, is formed
by an NTPDase that hydrolyzes ATP and ADP to AMP, 5-nucleotidase and adenosine deaminase (ADA) that hydrolyzes AMP
and adenosine respecively. Therefore, the aim of this study was to investigate the effect of aqueous leaves extract of S. buxifolia
on enzymes that hydrolyze adenine nucleotides in lymphocytes from rats.
Adult male Wistar rats (200-250 g) obtained from the Central Animal Facility from the Federal University of Santa Maria, RS,
Brazil, were utilized from experiments. Rats were euthanized and the lymphocytes were prepared for the NTPDase and ADA
activities assays (nmol Pi/min/mg of protein and U/mg of protein) according to (Diabetes Res. Clin. Pract. 65:1, 2004). The
enzyme preparation (8-12 g of protein) were added to the reaction mixture and pre-incubated at 370 C with extract of S.
buxifolia at concentrations of 10, 50, 100 and 200 g/mL. The results were analyzed by one-way ANOVA, followed by Duncans
multiple range test. P
Conclusions:
The medicinal plants have been extensively used to prevent and treat a broad variety of diseases and other medical conditions.
Circulating nucleotides are known to be important signalling molecules that render (potentially) a variety of physiological
responses. These results indicate that Scutia buxifolia can exert modulatory effect in the purinergig signaling in the immune
system increasing the ATP levels in the circulation. In this way, we propose that the decrease of NTPDase activity of
lymphocytes demonstrated in this work due to the action of chelating flavonoids present in leaves extract of Scutia buxifolia.
Additional studies are necessary to identify the active components of Scutia buxifolia that produce the effects on lymphocytes
with the intention of finding a better therapy to benefit patients with immune disorders.
Keywords: Scutia buxifolia , Adenosine deaminase , Lymphocytes, Enzymes
Financial Support: Capes; CNPq, FAPERGS
Resumo:31-062
PARTIAL ISOLATION OF PHOSPHOLIPASE A2 INHIBITORS PRESENTS IN THE CAUDISONA DURISSA
COLILLINEATUS (CROTALUS DURISSUS COLILLINEATUS) SERUM.
Gimenes, S. N. C. 1; Mendes, M. M. 1; Ferreira, F. B. 1; Silveira, A. C. P. 1; Gomes, M. S. R. 1; de Lima, L.

F. G. 1; Brites, V. L. C. 2; Santos, A. L. Q. 3; Rodrigues, V. M. 1
1
Instituto de Gentica e Bioqumica da Universidade Federal d, INGEB/UFU
2
Instituto de Biologia da Universidade Federal de Uberlndia, INBIO/UFU
3
Faculdade de Medicina Veterinria da Universidade Federal de, FAMEV/UFU
Objectives:
It is known that some animals have a natural resistance to the toxic effects of snake venom, which is caused by the presence of
endogenous inhibitors in the serum of these animals. The PLA2 inhibitors represent an important mechanism against the toxic
effects of snake venom. So this paper aims the isolation and the functional characterization of the PLA2 inhibitors from
Caudisona durissa colillineatus (Cdc) serum.
The phospholipasic A2 of different venoms and PLA2 toxin (BnSP7) from Bothrops pauloensiswas assayed by potenciometric
method by measuring the fatty acids released of a solution of egg yolk (n=3). The myotoxic activity was assayed by the plasma
creatine kinase level after 3h of injections in gastrocnemius muscle of Swiss male mice (n=4). For the inhibition tests, the venoms
or toxin were previously incubated with serum or inhibitors for the 30 min at 37C at ratios 1:5, 1:10 and 1:20 (venoms or toxin:
serum or inhibitors, w/w). The PLA2 and myotoxic activities of different venoms were efficiently inhibited by the serum of Cdc,
reaching values of 13% to 70% of inhibition. The Bothrops pauloensis venom activities were the most inhibited. The partial
isolation of inhibitors of Cdc serum was performed by two chromatographic steps. Firstly on the ion exchange Q-Sepharose Fast
Flow, resulting in six distinct peaks (Q1 to Q6). Q4 fraction inhibited 50% of phospholipase activity of B. pauloensis venom at
ratio 1:20 (w/w). This fraction was subsequently chromatographed on NHS affinity column activated with PLA2 (BnSP7),
resulting in two fractions called NHS-1 and NHS-2. NHS-2 inhibited 100% the phospholipasic activity induced by acid PLA2 at
a ratio of 1:5 (w / w), in contrast, the NHS-1 fraction was not able to inhibit such activity in any ratio assayed.
Conclusions:
Our results showed that Cdc serum and the NHS-2 fraction contain proteins able to inhibit the PLA2 activity of different venoms
and the BnSP7 toxin. Thus, this work opens up new perspectives for the application of inhibitors as a structural model for the
synthesis of compounds able to neutralize the toxic effect of snake venoms, as well as a potential medication for treatment of proinflammatory diseases arising from the action of phospholipases in cells.
Keywords: Crotalus durissus colillineatus, Inhibitor, serum of snakes
Resumo:31-063
CYTOTOXIC EFFECT OF BOTHROPS DIPORUS ON MDCK CELLS
Marinho, A. D. 1; Jorge, R. J. B. 1; Josino, T. S. 2; Menezes, R. R. P. P. B. D. 1; Morais, I. C. O 1; Guedes,

A. L. M. D. O. 1; Toyama, M. H. 3; Martins, A. M. C. 2; Santos, J. V. 1; Monteiro, H. S. A. 1
2
Departamento de Anlises Clnicas e Toxicolgicas, DACT-UFC
1
Departamento de Fisiologia e Farmacologia, DFF- UFC
3
Universidade Estadual Paulista, UNESP
Objectives:
We have studied the cytotoxic effect of the venom of B. diporus (VBd) on renal tubular cells Mardin-Darby Canine Kidney
(MDCK).
The MDCK cells were cultured in RPMI medium supplemented with 10% fetal bovine serum and penicillin / streptomycin, and
incubated in a 5% CO2 at 37 C for 3-5 days until reaching confluence state, and it was kept always aseptic conditions. Then,
they were displaced with trypsin-EDTA (0.05%/ 0.02%), counted in a Neubauer chamber and plated to 1x105 cells/mL in 96well plates. After 24h of incubation, the plate was washed with PBS and different concentrations (100, 50, 25, 12.5, 6.25, 3.12, 1,
56, 0, 78 and 0,39g/mL of VBd was added. Then, it was added to 10L of MTT solution (2.5 mg/mL), and 4h later, 90L of
sodium dodecyl sulfate (SDS) at 10% was also added. The spectrophotometric reading at 570nm was performed 17h later. As a
negative control PBS was used. The results were expressed as percent viability SEM in the control group. Three experiments
were conducted in triplicate, and data were analyzed with GraphPad Prism 5.0, by ANOVA with Dunnet post-test, with
significance at p B. diporus (VBd) was able to decrease cell viability under the study conditions, by showing cytotoxicity effect
up to 0,78 g/mL (IC50 = 1,09 g/mL and promoting an inhibition on the cell growth in a dependent-concentration.
Conclusions:
Other studies, with fractions of VBd will be conducted to ascertain which is the fraction involved in the role of the toxic effects in
MDCK culture cells and also to investigate the mechanisms involved in cell death.
Keywords: MDCK, Bothrops diporus , CYTOTOXIC
Financial Support: CNPq; CAPES; FUNCAP
Resumo:31-064
PROTECTIVE EFFECT OF MORPHINE AGAINST MERCURY INTOXICATION IN VITRO: INVOLVENMENT OF
OPIATE RECEPTORS?
Malaquias, A. C. ; Almeida, M. B. ; Nascimento, J. L. M. D. ; Maues, L. A. L. ; Herculano, A. M. ; Crespolpez, M. E.

Instituto de cincias biolgicas, UFPA
Objectives:
Morphine is a potent opioid analgesic used as a major therapeutic option for treatment of moderate to severe pain, especially of
oncologic nature. However, other possible effects of morphine, as in oxidative stress and mercurial intoxication, have been
highlighted recently. Methylmercury (MeHg) is one of the most dangerous environmental contaminants and its target organ is the
central nervous system. In Amazon, accumulation of this heavy metal causes mercury levels in humans above the limits
recommended by the World Health Organization. Oxidative stress is one of the major mechanisms underlying the deleterious
effects of mercury intoxication. Recently, an antioxidant role for morphine was demonstrated, being a hypothesized receptorindependent effect Almeida et al., 2010 (Dissertation PPGNBC 2010, UFPA). Thus, the purpose of this work was to show the
possible influence of morphine in C6 glioma cells exposed to MeHg evaluating cellular viability and the involvement of opiate
receptors.
C6 glioma cells were grown at 37C and 5% CO2 in DMEM with 10% fetal bovine serum (FBS), 100 U/ml of penicillin and
100g/ml of streptomycin. Cellular viability was tested by MTT assay as described by Mosmann et al. (Jorn. Immunol. Meth. 65,
55,1983). One-way ANOVA test, following by post hoc Tukey test when appropriated, was used (significant values for p were
set at 0.05). The results were expressed as mean standard deviation of percentage of the control group (n=4). Cell viability
decreased in a dose-dependent way when treated with 0-11 M of MeHg for 24 hours. A DL50 of 6.97M of MeHg was
observed. Exposure to morphine (0-10M), naloxone (0-2.5M), or co-treatment with morphine and naloxone (1 and 2.5 M,
respectively) did not cause significant differences in cell viability with that of control group. Exposure to 6M of MeHg reduced
cell viability to 58.94 3.66% of the control group. Interestingly, co-treatment with morphine 1 M increased the viability to
74.1 2.24% of the control group. To test whether this protective role of morphine was via opioid receptors activation, the major
antagonist of these receptors, naloxone (2.5M), was added. Surprisingly, the protective effect of morphine against mercury
intoxication was completely eliminate by naloxone, reducing the viability to 62.91 5,29%.
Conclusions:
This work demonstrates, for the first time, that therapeutic doses of morphine significantly protects against cell death caused by
6M MeHg intoxication (brain concentrations of 2.5-10M of MeHg have been associated with delayed psychomotor
development in children and adults). The prevention of the morphine protective role by the antagonist opioid receptors, naloxone,
implies that the protective effect on cell viability caused by morphine could be attributed to the action of morphine on opioid
receptor (MOR mainly). Thus, more studies on the MOR pathways of signal transduction must be carried out in order to establish
the exact molecular mechanism of the protective effect of morphine against mercury intoxication.
Keywords: morphine, oxidative stress, mercury, naloxone, c6 cells
Financial Support: CAPES, CRISTLIA, CNPq
Resumo:31-065
MOTOR DAMAGES AND BRAIN LEAD BIOACCUMULATION CAN BE ATTENUATED BY GALLIC ACID
TREATMENT IN LEAD-EXPOSED RATS
Reckziegel, P. 1; Dias, V. T. 1; Benvegn, D. 1; Boufleur, N. 1; Barcelos, R. C. S. 1; Segat, H. J. 1; Pase, C.

S. 1; Santos, C. M. M. D. 2; Flores, . M. M. 2; Brger, M. E. 1
1
Departamento de Fisiologia e Farmacologia, UFSM
2
Objectives:
The present study investigated the effects of gallic acid (GA), a natural compound found in vegetal sources, on brain
bioaccumulation and motor damages induced by lead in rats.
Rats were exposed to lead nitrate (50mg/Kg, ip) or saline (1 mL/Kg, ip) by 5 days. From day 6, the rats were treated with GA
(13.5 mg/Kg/mL, po), EDTA (110 mg/Kg/mL, ip) or water (1mL/Kg, po) by 3 days. Two hours after the last dose, crossing and
rearing numbers were quantified in open-field test. In addition, fifteen hours after the last dose, the animals were anesthetized
(sodium thiopental - 50mg/Kg, ip) and euthanized by exsanguination. Accumulation of lead was measured in brain using
inductively coupled plasma optical emission spectrometer. Animals were maintained and used in accordance with the guidelines
of the National Council for Control of Animal Experiments (CONCEA) (protocol number 109/2010). Data were analyzed by oneway ANOVA, followed by Duncans multiple range test. Correlations were investigated by Pearsons correlation coefficient.
PF(4,30)= 8.11 and 9.34, PF(4,20)=23.89; PPP
Conclusions:
The GA, a polyphenol compound often found in vegetables sources, was able to partially reverse these decreases in motor
activities (crossing and rearing) in the lead-exposed animals. Interestingly, the standard treatment with EDTA did not change
these lead-induced motor damages, thus raising questions regarding its beneficial effects when used alone in lead poisoning.
These decreased of locomotor and exploratory activity are associated to lead brain accumulation, according to correlation results.
GA benefits may be partially by its chelating property showed here and by its high antioxidant potential.
Keywords: gallic acid, lead, motor damages
Financial Support: CNPq, CAPES
Resumo:31-066
EFFECTS OF PAULLINIA CUPANA EXTRACT SUPPLEMENTATION ON CRYOPRESERVATION MEDIUM OF
HUMAN SEMEN AND LEUCOCYTES
Cadon, F. C. 1,1,1,1; Flres, E. R. S. 1,1,1,1; Fantinel, M. 1,1,1,1; Mnica-cattani, M. F. 1,1,1,1; Montagner, G. F.

D. S. 1,1,1,1; Jobim, M. 1,1,1,1; Algarve, T. D. 1,1,1,1; Ribeiro, E. E. 2; Garcia, L. F. M. 1; Cruz, I. B. M. D. 1,1,1,1
1
Universidade do Estado do Amazonas, UEA
Objectives:
To analyze oxidative stress markers after the cryopreservation of human semen and leucocytes using hydro-alchoolic extract of
Paullinia cupana (guaran) frozen medium supplementation.
We performed an in vitro prospective study using semen and leucocytes from a fertile and healthy men analyzing if the guaran
suplementation decrease the oxidative stress caused by cryopreservation. Reactive oxygen species (ROS) with 2',7'dichlorofluoresceinacetate (DCFH-DA) determined by fluorometry and lipid peroxidation (TBARS) determined by
spectrophotometry as well as viability cells (MTT assay) were analyzed in samples treated with 0, 1, 5, 10 and 20 g/mL of
guaran. The guaran supplementation showed post-thawing protection on oxidative stress variables and viability in the samples
analyzed here. The best concentration of the guaran supplementation was 10 g/mL then negative control group and other
concentrations.
Conclusions:
Guaran supplementation positively affects the condition of the viability and oxidative stress of cells cryoconserved. The results
indicated that the concentration of 10 g/ml of guaran as the best dose of supplementation. Further studies on the viability of the
sperm should be conducted.
Keywords: Paullinia cupana , CRYOPRESERVATION, HUMAN SEMEN , LEUCOCYTES, OXIDATIVE STRESS
Resumo:31-067
CYTOTOXIC EFFECTS OF ONCOCALYXONE A AGAINST HUMAN PROMYELOCYTIC LEUKEMIA CELL
LINE HL-60
Sbardelotto, A. B. 1; Barros, F. W. A. 1; Cabral, I. O. 1; Rocha, D. D. 1; Meira, A. S. 1; Moraes, M. O. D. 1;

Rodrigues, F. A. R. 1; Pessoa, O. D. L. 2,2; Cavalcanti, B. C. 1; Pessoa, C. D. O. 1
1
Departamento Fisiologia e Farmacologia, UFC
2
Departamento Qumica Orgnica e Inorgnica, UFC
Objectives:
Auxemma oncocalyx Taub. belongs to the Boraginaceae family. It is known as pau branco and is frequently found in the State
of Cear (Northeastern Brazil). The bark of the tree is an astringent and is popularly used in the treatment of wounds.
Oncocalyxone A (Onco-A), a quinone isolated from the EtOH extract of A. oncocalyx, exhibited a series of pharmacological
properties, such as analgesic, anti-inflammatory, antioxidant, cytotoxic and antitumor properties. The present study was
undertaken to provide a basic set of data on the cytotoxic effects of Onco-A against human promyelocytic leukemia cell line HL60.

The cytotoxic potential of Onco-A (0.8 to 5 g/mL) was evaluated by the MTT assay after 24 h exposure. In order to determine
the mechanisms involved in its cytotoxic effects, cells were treated with increasing concentrations (2.5, 5 and 10 g/mL) of
compound during 24 h. After cells exposure, flow cytometry experiments (cellular viability and internucleosomal DNA
fragmentation) and morphological analysis with DNA-binding fluorescent dye (acridine orange/ethidium bromide) were
performed. Moreover, to determine whether reactive oxygen species (ROS) are involved with Onco-A-induced cell death, we
examined the intracellular generation of ROS by flow cytometry using a fluorescent probe (H2DCFDA) able to detect different
oxygen radicals such as hydroxyl, peroxyl radicals, and peroxynitrite anion. Based on results of MTT test, Onco-A showed a
significant cytotoxic activity against HL-60 cells (IC50 = 3.7 0.4 g/mL). In addition, flow cytometry and morphological
analysis of cells treated with lower concentration of Onco-A, demonstrated a reduction in cell volume, formation of apoptotic
bodies, chromatin condensation, reduction of cell viability and DNA fragmentation (sub-diploid sized DNA). Also, our results
showed that ROS do not play an important role on Onco-A-cytotoxicity towards HL-60 cells.
Conclusions:
These findings suggest that apoptosis is an important form of cell death caused by Onco-A, and reinforce that this compound
might be potential lead prototypes for the cancer chemotherapy.
Keywords: apoptosis, cancer, cytotoxic, Oncocalyxone A
Financial Support: CNPq, CAPES, FUNCAP, BioTechCell
Resumo:31-068
NEUROPATHOLOGICAL AND BEHAVIORAL EFFECTS OF THE EXPERIMENTAL ALUMINUM CITRATE
INTOXICATION IN ADULT RATS.
Silva Jnior, A. F1; Aguiar, M. S. S. 1; Carvalho Junior, O. S. 1; Feio, R. A. 2; Faro, L. R. F. 3; Lima, R. R. 4;

Gomes-leal, W. 4
1
Laboratrio de Psicobiologia/Universidade Federal do Par, UFPA
3
Laboratrio de Neurotoxicologia/Universidade Federal do Par, UFPA
2
Laboratrio de Farmacologia/ Universidade Federal do Par, UFPA
4
LNNE/Universidade Federal do Par, UFPA
Objectives:
Experimental evidences suggest that aluminum is a neurotoxic agent with deleterious actions on cognitive processes.
Nevertheless, few studies have systematically investigated both neurobehavioral and neuropathological effects of the
experimental intoxication with aluminum. In this study, we have investigated the neurobehavioral effects of the experimental
contamination with aluminum citrate (AC) on the mnemonic processes of adult Wistar rats.
29 adult male Wistar rats weighing 230-250 g, were divided into one control group (G1) and 3 AC-treated groups (G2, G3 and
G4) with survival times of 8, 17 and 31 days, respectively. The neurotoxicant (320 mg/kg) was intragastrically administered for 4
days. Behavioral analysis was assessed using both open field and elevated T-maze tests. Animals were perfused with 0.9% saline
solution and 4% paraformaldehyde. Neuronal loss (anti-neuN) and astrocyte activation (anti-GFAP) were assessed in coronal
sections containing CA1 and CA3 hippocampal regions using immunohistochemistry. There was an increase in the locomotor
activity in open field test for G2 in comparison to control and other groups (P
Conclusions:
The results suggest that the experimental intoxication with AC induces hippocampal damage with associated learning and
memory deficits. This supports the hypothesis that pathological accumulation of Aluminum may impair mnemonic hippocampal
processes.
Keywords: Rat, Aluminium Citrate, Hippocampus, Memory, Learning
Financial Support: Foundation of Research and Development of the State of Para
Resumo:31-069
RLOSAC, A RECOMBINANT HEMOLIN, INDUCES CELL SURVIVAL THROUGH REGULATION OF CELL
CYCLE AND MITOCONDRIAL ACTIVITY
Bosch, R. V. ; Alvarez-flores, M. P. ; Vaz-de-lima, B. B. ; Maria, D. A. ; Chudzinski-tavassi, A. M.

Bioqumica e Biofsica/Instituto Butantan, IB
Objectives:
Losac is a 45-kDa protein from Lonomia obliqua caterpillar that promotes blood coagulation through specific proteolytic
activation of factor X. In cultures of HUVECs, Losac increased cell proliferation and inhibited the apoptosis induced by
starvation (Biochem. Biophys. Res. Commu. 343; 1216, 2006). Recombinant Losac (rLosac) was recently obtained from bacteria
system (J. Biol. Chem. 286; 6918, 2011). In this work, we investigated the effect of rLosac on endothelial cells (HUVECs) in
quiescent (10% FBS) and serum deprivation (1% FBS) conditions
Human umbilical vein endothelial cells (HUVECs) were obtained by digestion of umbilical cord veins with collagenase as
described previously (J. Clin. Invest. 52, 2745, 1973). Cell cycle phases, apoptosis/necrosis death (Anexin-V/PI), expression of
phosphorylated caspase 3 and mitochondrial electric potential (Rhodamine-123) were analyzed by flow cytometry. HUVECs
(n=03) were treated with 5, 10 and 20nM rLosac in the presence of 1% or 10% FBS during 48 and 96 hours. Results: There was
no changes in the proportion and distribution of cells in different cell cycle phases rLOSAC treated group compared to control,
with 1% FBS. HUVEC's treated with different concentrations of rLosac in the presence of 10% FBS and after 48 and 96 hours
showed a decrease of cells with fragmented DNA (36.9%) and the activity of caspase 3 (control 45.2%; rLosac 17.2%) (p<0.05).
Conclusions:
Under these conditions it is evident the increase in metabolic activity of mitochondria independently of caspase-3, showing its
effect as extrinsic regulator of cell survival.
Keywords: Lonomia obliqua , Losac, Coagulation, HUVEC, Factor X activator
Financial Support: FAPES, CAPES, CNPQ, CAT-CEPID
Resumo:31-070
HUMAN NEUROBLASTOMA CELLS TRANSFECTED WITH TYROSINE HYDROXYLASE GAIN INCREASED
RESISTANCE TO METHYLMERCURY-INDUCED CELL DEATH
Franco, J. L. 1; Paula, M. T. D. 2; Zemolin, A. P. P. 1; Salvad, V. S. 1; Rodrigues, N. R. 1; Posser, T. 1

1
Campus So Gabriel, Universidade Federal do Pampa, UNIPAMPA
2
Objectives:
In a previous study we demonstrated that human neuroblastoma SH-SY5Y cells transfected with human tyrosine hydroxylase
isoform 1 (SH+TH cells) were substantially more resistant to cell death induced by pro-oxidants than wild type SH-SY5Y cells
(SH cells). In the present communication we used methylmercury as a model of cell stress in order to test whether SH+TH cells
would behave in a similar manner in response to this stressor (Toxicol. in Vitro 24: 1498, 2010).
Experiments were performed using wild-type human neuroblastoma SH-SY5Y cells obtained from the American Type Tissue
Culture (http://www.atcc.org), or SH-SY5Y cells transfected with human TH isoform 1 (Toxicol. Sci. 113(1): 150, 2010). Cells
were incubated with methylmercury (0.13 microM) for 24 h and then, analysis of cell viability (MTT test), apoptotic markers (%
apoptotic nuclei, poly(ADP ribose polymerase cleavage) and p38MAPK phosphorylation was evaluated. Methylmercury caused a
signicant reduction in cell viability and increased apoptotic markers in both cell types. This Effect was dependent on
concentration of MeHg tested. However, the effects were significantly reduced (p
Conclusions:
In conclusion our results show that insertion of the human TH gene in cells that originally do not express this protein leads to
alterations in cell homeostasis and triggers defense mechanisms against pro-oxidative insults.
Keywords: Tyrosine hydroxilase, SH-SY5Y, transfection, cell death
Financial Support: CNPq, FAPERGS.
Resumo:31-071
BIOCIDE EFFECT OF AN ANTARCTIC ALGAE (PRASIOLA CRISPA) EXTRACT IN THE ADULT FRUIT FLY
DROSOPHILA MELANOGASTER.
Zemolin, A. P. P. ; Nunes, M. E. M. ; Pereira, B. K. ; Pereira, A. B. ; Belo, C. A. D. ; Posser, T. ; Franco, J.

L.
UNIVERSIDADE FEDERAL DO PAMPA, UNIPAMPA
Objectives:
The insecticidal properties of a number of plants have been investigated for thousands of years, and some of the plants can
substitute many synthetic means of control. In this respect, it is important to emphasize that natural agents are environmentally
less harmful than synthetic pesticides. The aim of this study was to evaluate the effects of the ethanolic extract of the terrestrial
eukaryotic green alga from Antarctica, Prasiola crispa on survival of adult D.melanogaster, and in parallel, to verify a possible
modulation of antioxidant enzymes activity and locomotor performance in response to the exposure of this organism to the
Prasiola crispa extract. The fruit fly Drosophila melanogaster, belongs to the order Diptera and family Drosophilidae. This model
is recognized for its high sensitivity to toxic substances, thus being considered a bioindicator for detection of pollutants and also
to test the biological action of natural substances.
Toxicity was assessed as mortality rate (fold increase), negative geotaxis behavior and acetylcholine esterase (AchE), glutathione
S-transferase (GST), catalase (CAT) activities as well as glutathione content (GSH) and hydroperoxide formation. Administration
of algae extract (2 mg/ml diluted in sucrose 2%) to flies for 24 hours resulted in a massive increase in mortality (7.6 fold increase
(p
Conclusions:
Our results show for the first time the toxic effects of an Antarctic algae extract in Drosophila melanogaster. The insecticide
action of Prasiola crispa may be related to changes on key cell redox-active systems. Further studies are ongoing in our laboratory
in order to elucidate the exact mechanisms of toxicity of this Antarctic algae to Drosophila melanogaster.
Keywords: ANTARCTICA, BIOPESTICIDE, DROSOPHILA MELANOGASTER, PLANT EXTRACT, TOXICITY
Financial Support: PBDA Unipampa; INCT-APA; CNPq; FAPERJ; FAPERGS
Resumo:31-072
EXPOSURE TO AIR POLLUITION INCREASES THE DELETERIOUS EFFECTS WITH ADVANCING AGE?
Domenico, M. D. 1; Ciccone, A. C. 1; Fagundes, L. S. 1; Zanchi, A. C. T. 1; Saldiva, P. H. N. 2,1; Rhoden, C.

R. 1
1
2
Objectives:
Studies have shown that pollution from the city of Porto Alegre cause oxidative stress in the cortex of rats exposed to pollutants
during the pre-and postnatal, as well as impairs short-term discriminative memory (Inhal Toxicol. 910-8, 2010). However, it was
not analyzed the issue of age in this study. The aim of this study was to evaluate if exposure to air pollution in animals with 45
days and 90 days of age changes behavior with advancing age. In addition, it was evaluated the effect of pollution on oxidative
stress and endogenous antioxidants of cerebral cortex.
The rats (n=36) were evaluated in different stage of development, 45 and 90 days, by morris water maze that analyzed the spatial
memory in the animals (Physiol.Behav. 781-785, 1992). Besides, it was analyzed oxidative stress in the cortex by superoxide
dismutase activity measure (SOD). Statistical Analysis were performed by one-way analysis of variance (ANOVA) followed by
the Post hoc multiple comparisons Student NewmanKeuls test. The level of significance was set at 5%. The analyze of morris
water maze, which evaluate the identification spatial time, showed a statistic difference among 90 days nonfiltered environment
animals in comparison with the other groups: 45 days filtered (p=0.001); 45 days nonfiltered (p
Conclusions:
These results suggest that there is damage in older animals in relation to spatial memory according to the animal is exposed to the
polluted environment. Furthermore, became evident, by decreasing the activity of the SOD, that exposure to pollution affects the
cortex by the oxidative pathway.
Keywords: behavior, oxidative stress, pollutants
Financial Support: Universidade Federal de Cincias da Sade de Porto Alegre
Resumo:31-073
EFFECT OF SUBACUTE ADMINISTRATION OF (S)-DIMETHYL 2-(3-(PHENYLTELLANYL) PROPANAMIDO)
SUCCINATE ON OXIDATIVE STRESS PARAMETERS OF MICE
Meinerz, D. F. 1; Comparsi, B. 1; Mariano, D. O. C. 1; Santos, D. B. 2; Allebrandt, J. 1; Prestes, A. S. 1;

Stefanello, S. T. 1; Franco, J. L. 3; Rocha, J. B. T. 1
1
2
3
Objectives:
Previous studies demonstrated that organotellurium compounds (OTeC) are potentially toxic and lethal to rodents at low doses.
OTeC toxicity can be related to their interaction with thiol groups of important biomolecules and to the induction of reactive
species by OTeC. In this context, we evaluated the potential toxic effect of (S)-dimethyl 2-(3-(phenyltellanyl) propanamido)
succinate (TeAsp), a telluroamino acid derivative of aspartic acid, and the possible protection effect of N- acetylcysteine (NAC),
a thiol-containing antioxidant.
Swiss adult male mice weighing 30-40 g, were used in accordance to guidelines of the Committee on Care and use of
Experimental Animal Resources of the UFSM, Brazil (23081.002435/2007-16). Mice were divided in four groups (n=5). The
animals were treated by 10 days. In the first 3 days of treatment mice received one daily injection of NAC (100mg/kg) or
phosphate buffer (PB) 2.5ml/kg (i.p.). After de third day, mice received either one of these and, also by the next 7 days injections
of TeAsp (92.5 mol/kg) or DMSO (vehicle) 1ml/kg (s.c.), resulting in the following groups: (1) PB+ DMSO; (2) NAC +
DMSO; (3) PB+ TeAsp; (4) NAC+ TeAsp. Mice were weighted daily. Exploratory and motor activity was determined in an
open-field (OF) apparatus, 24 hours after the last injection of treatment. Mice were decapitated and liver and brain were removed
for quantification of catalase (CAT), glutatione peroxidase (GPx) and reductase (GR) and thioredoxin reductase (TrxR). Data are
expressed as Mean S.E.M. Statistical analysis was performed using one-way analysis of variance (ANOVA), followed by
Newman-Keuls multiple comparison test. Results were considered statistically signicant when p < 0.05. TeAsp caused a marked
reduction in weight gain of mice (p
Conclusions:
In contrast to our expectation, NAC could not protect mice from the toxicity of TeAsp. In effect, NAC exacerbated the general
and behavioral toxicity of TeAsp (body weight loss and OF activity). Furthermore, association of NAC+TeAsp increased hepatic
GR. Although not determined here, the exacerbation of TeAsp toxicity by NAC can be associated with an acceleration of TeAsp
metabolism to a more toxic intermediate (for instance, diphenyl ditelluride) or to release of Te from the organic moiety.
Keywords: Tellurium compound, toxicity, oxidative stress, mice, NAC
Resumo:31-074
IN VITRO NEPHROTOXICITY OF BOTHROPS LEUCURUS VENOM .
Morais; I. C. O1; Dantas, R. T. 1; Torres, A. F. C. 2; Jorge, A. R. C. 1; Marinho, A. D. 1; Jorge, R. J. B. 1;

Guedes, A. L. M. D. O. 1; Toyama, M. H. 3; Monteiro, H. S. A. 1; Martins, A. M. C. 2
1
2
Departamento de Anlise Clnicas, FFOE, UFC
3
Departamento de Bioqumica, UNICAMP
Objectives:
The aim of this study was to investigate the effects of the Bothrops leucurus venom in the renal perfusion system and in cultured
renal tubular cells of the type MDCK (MadinDarby canine kidney.
Isolated kidneys from Wistar rats weighing 250 to 300g (n=6) were perfused with Krebs-Henseleit solution containing 6% of
bovine serum albumin previously dialyzed for 120 minutes. The effects of Bothrops leucurus venom (VBBL) (10g/mL) were
studied on glomerular filtration rate (GFR), urinary flow (UF), perfusion pressure (PP), renal vascular resistance (RVR) and
percentage sodium (%TNa+), potassium (%TK+) and chloride (%TCl-) tubular transport at 60, 90 and 120 minutes of
experiment. All data were analyzed by unpaired t test with level of significance of *pBothrops leucurus venom in the
concentrations of 100, 50, 25, 12.5, 6.25, 3.12 and 1.56 g/mL 24 hours afterwards, cytotoxity was available by assay MTT,
with detects cell viability on the basis of oxidative metabolism. Three experiments were conducted in triplicate, and data were
analyzed with GraphPad Prism 5.0, by ANOVA with Dunnet post-test, with significance at p B. leucurus venom (10 g/mL)
reduction the PP at 90 and 120 min (PPCT=110.2 4.3; PP90= 84.9 5.0* mmHg; PP120= 78.9 3.9 mmHg) and UF at
60 and 90 min (UFCT=0.134 0.009; UF60= 0,114 0.01*; UF90= 0.073 0.01 mL/g-1.min-1). The glomerular filtration
rate decreased at 60 and 90 min (GFRCT=0.701 0.064; GFR60=0.425 0.047*; GFR90=0.199 0.036* mL/g-1.min-1).
The renal vascular resistance decreased at 120 min (RVR CT= 5.39 0.51; RVR120 =4.13 0.17*). It was also observed a
decrease on percentual tubular transport of sodium (%TNa+) at 120 min and of chloride (%TCl-) at 60 and 90 min
(%TNa+CT=81.94 1.05; %TNa+90=64.54 2.44*); (%TCl-CT=79.90 2.00; %TCl-60= 71.34 2.58 *; %TCl-90=
61.74 1.89*). Treatment with B. leucurus venom significantly decreased is viability on MDCK cells at all concentrations
tested with an IC50 of 1.25 g/mL.
Conclusions:
The Bothrops leucurus venom caused nephrotoxicity in isolated kidney and cytotoxicity in MDCK cells.
Keywords: Botrhops leucurus, Kidney, MDCK, Toxicity
Financial Support: CNPQ; FUNCAP
Resumo:31-075
EXPERIMENTAL ENVENOMATION INDUCED BY BOTHROPS JARARACUSSU (VIPERIDAE, CROTALINAE)
SNAKE VENOM: EVALUATION OF THE EFFICACY OF TREATMENT USING DIFFERENT ANTIVENINS
Yano, M. Y. ; Sano-martins, I. S.
Laboratrio de Fisiopatologia, IBu
Objectives:
The aim of this study was to compare the efficacy of different antivenin treatments in the experimental envenomation induced by
Bothrops jararacussu (Bju) snake venom by the evaluation of clot parameters by thrombelastography (TEG), and fibrinogen
levels.
Groups of mice (n=5) were injected intravenously with 2 x Minimum Defibrinogenating Dose (0,50mg/kg) of Bju venom, or
saline (envenoming control), and treated with botropic (SAB), crotalic (SAC) and botropic/crotalic (SABC) antivenin, or saline
(treatment control), 1 hour (h) after venom inoculation (i.v.). Blood samples were collected in 3, 6, and 12 hs after the treatment,
by the orbital plexus. The plasma fibrinogen levels (PFL) were determined 3, 6 and 12 hs after the treatment. TEG was carried
out on whole blood samples with the ROTEM coagulation analyzer, according to the standard protocols supplied by the
manufacturer. The NATEM test was used to assess whole blood clot formation in the absence of activation of clotting cascade
other than the recalcification and spontaneous contact activation. Test parameters as Clotting time (CT, seconds), Clot Formation
Time (CFT, seconds), Alfa-angle () and Maximum Clot Firmness (MCF, mm) were acquired for 1 h. Statistical analysis were
accessed by one-way ANOVA and Tukey test. Regarding PFL at 3 h, the animals envenomed and treated with different
antivenom showing huge variation in the groups. And at 12 h there were no statistical difference of amount of fibrinogen between
the groups treated with saline or antivenin, indicating that fibrinogen was spontaneously normalized independently of the
treatment. On the other hand, at 6 hours, the animals treated with SABC showed significantly (p
Conclusions:
Animals envenomed by Bju and treated with SABC at 6hs, showed significantly higher levels of fibrinogen when compared to
those treated with SAB, SAC and saline. The analysis of parameters obtained by TEG corroborates these results, indicating that
treatment with SABC recovers faster than SAB and SAC.
Keywords: Antivenenos, Bothrops jararacussu, Envenenamento, Soroterapia
Financial Support: INCTTOX, CNPq and CAPES
Resumo:31-076
HEMATOLOGICAL CHANGES IN PROCHILODUS LINEATUS EXPOSED TO PHENANTHRENE.
Martins, L. P. A. ; Ferreira, J. D. F. ; Fernandes, M. N.

Departamento de Cincias Fisiolgicas, UFSCar
Objectives:
Petroleum and its derivatives are potential pollutants in aquatic ecosystems which may affect the biota, including the fish living in
such environment. This study evaluated the effects of acute exposure to phenanthrene on the hematological variables of
curimbat, Prochilodus lineatus, species widely distributed in Southeastern and Southern Brazil and characterized as a potential
vertebrate bioindicator of water quality in freshwater systems.
Fish (n = 12 in each test, Wt = 39.0 3.1 g) were divided into four experimental groups: control group (C) and groups exposed to
10 g/L (E1), 20 g/L (E2) and 200 g/L (E3) of phenantrene for 96 hours. After exposure, a sample of blood was taken from the
caudal vein using heparinized syringes for analyzing the hematological variables. After applying the D'Agostino & Pearson test
for data normality analysis were used ANOVA one way followed by Bonferroni post-test at P>0.05 significant level to compare
data. Red blood cells (RBC) number (erythrocytes number/L) and hematocrit (%) were not significantly different between the
control and exposed groups (p>0.05). Hemoglobin concentration was higher in E1 (5.3 0.24 g/dL) and MCHC (mean
corpuscular hemoglobin concentration) was higher in E2 (14.4 0.8 g/dL) compared to control (12.2 0.3 g/dL). The MCV
(mean corpuscular volume) (138.4 5.2 m3) and MCH (mean corpuscular hemoglobin) (19.4 1.34 pg/cell) were higher in fish
from E3 group than in the controls (MCV = 111.0 6.5; MCH = 13.3 1.0). Total leukocytes decreased in E1 (61612 4745)
and E3 (70209 8253) groups compared to controls (107074 11211) while the thrombocytes did not show significant
difference. Among the leukocytes, the lymphocytes increased significantly in fish from groups E2 (97.57 0.39%) and E3 (97.17
0.77%). Erythrocyte nuclear abnormality in the phenantrene exposed fish was not significant different from controls varying
from 0.6 to 0.9% but, the percentage of micronuclei increased from 0.003 0.01 % in the controls to 0.037 0.01 % in E3.
Plasma K+ (6.2 0.23 mEq) and Cl- (107.9 5.07 mEq) concentration did not differed from controls but osmolality (229.7 6.1
mOsm) and Na+ (128.4 2.8 mEq) increased in E3 group (259.4 4.7 mOsm, Na+ = 145.1 3.3 mEq).
Conclusions:
The hematological results suggested that the changes observed in the red blood cells favor the increasing of oxygen uptake and
transport by the erythrocytes probably to overcome the increase of metabolic demand. The decrease in total leukocyte number
reduces the fish defense against pathogens in water and the lymphocyte increasing may be related to inflammatory processes
probably caused by the contaminant. Although micronuclei percentages were low, the increasing percentage in fish from E3
group suggests that phenantrene exposure to higher concentrations may have a mutagenic effect. The changes in plasma
osmolality and Na+ suggested ionic unbalance.
Keywords: FISH, HEMATOLOGY, PHENANTRENE, PROCHILODUS LINEATUS, TOXICOLOGY
Financial Support: CNPq/INCT-TA, CAPES, FAPESP
Resumo:31-077
FORMALDEHYDE INDUCES LUNG INFLAMMATION BY AN OXIDANT AND ANTIOXIDANT ENZYMES
MEDIATED-MECHANISM IN LUNG TISSUE
Lino-dos-santos-franco, A. 1; Vitoretti, L. B. 1; Correa-costa, M. 2; Dos-santos-duro, A. C. C. 3; Oliveira-
filho, R. M. 1; Saraiva-cmara, N. O. 2; Macourakis, T. 3; Tavres-de-lima, W. 1

1
Department of Pharmacology, ICB-1
2
Department of Immunology, ICB--4
3
Department of Clinical and Toxicological Analyses, FCF
Objectives:
Exposure to air pollutants such as formaldehyde (FA) leads to inflammation, oxidative stress and immune-modulation in the
airways. FA is emitted by building materials, furniture carpets, plywood, floor coverings and sterilizing agents. We recently
demonstrated that FA effects on the airways are partially due to an increased oxidative stress. However, the source of reactive
oxygen species (ROS) remained to be determined. Thus, we investigated the activity and gene expression of antioxidant and
oxidant enzymes after FA exposure.
Male Wistar rats (180 g) were exposed to FA inhalation (1%, 90 min daily) for 3 consecutive days. After 24 h of the last FA
inhalation, the activities and gene expression of glutathione peroxidase (GPX), glutathione redutase (GR), glutathione Stransferase (GST), superoxide dismutase (SOD) 1 and 2, catalase (CAT), inducible nitric oxide synthase (iNOS) and
cyclooxygenase (COX)- 1 were determined in the lung tissue. Data (mean SEM of 5 animals per group) were analysed by
ANOVA followed by the Student Newman-Keuls post test or the Students tailed paired t-test, whenever appropriate. A 4.0
version of the GraphPad InStat software was used for this purpose. P
Conclusions:
Our results indicate that FA causes disruption of the physiological balance of oxidant and antioxidant enzymes in lung tissue,
most probably favoring the oxidant pathways and thus positively modulating lung inflammation.
Keywords: FORMALDEHYDE, LUNG INFLAMMATION, OXIDANTS ENZYMES, ANTIOXIDANTS ENZYMES
Financial Support: FAPESP (2008/50766-1)
Resumo:31-078
INFLUENCE OF METAL IONS ON THE PROTEOLYTIC ACTIVITY OF A DIGESTIVE ENZYME FROM THE
VISCERA OF STRIPED MOJARRA (EUGERRES BRASILIANUS CUVIER, 1830)
Andrade, D. H. H. 1; Silva, J. F. 1; Nascimento Jr, C. R. 1; Silva, J. F. X. 1; Marcuschi, M. 1; Cah, T. B. 1;

Dantas, D. M. M. 1; Souza, K. S. 1; Ribeiro, K. 2; Bezerra, R. S. 1
1
2
Unidade Acadmica Especializada em Cincias Agrrias, EAJ/UFRN
Objectives:
Eugerres brasilianus is a fish that has an important role in estuarine fish fauna and of great importance to subsistence fishing in
the Northeast region of Brazil. Like other aquatic organisms, it is also subject to contamination by heavy metals originating from
anthropogenic activity, such as domestic sewage and effluents from industrial and agricultural production. The metals, in contact
with the organisms, can cause damage in their physiology, particularly in the enzymatic activity, compromising their
development. Based on this information, the aim of this study was to evaluate the proteolytic activity of trypsin from the pyloric
caeca and intestine of E. brasilianus in the presence of different metal ions.
Initially, the pyloric caeca and intestines were collected from seven specimens of E. brasilianus obtained from a fishing
community located in Itapissuma (PE - Brazil). The specimens had a average length and weight of 22.50.6 cm and 2107 g,
respectively. Then the organs were homogenized separately in 0.01M Tris-HCl, pH 0.8 with 0.9% NaCl (w/v) to obtain the crude
extracts (CE) of tissues. Using BApNA as specific substrate for trypsin, enzyme activity was performed in the presence of
different metal ions in the forms of AlCl3, BaCl2, CaCl2, CuCl2 and HgCl2 at concentrations of 0.5 mM, 2.5 mM and 5.0 mM.
The enzyme activity obtained in the absence of metals was taken as being 100% (control) and all tests were performed at 25 C
and pH 8.0. All data was analyzed using one-way analysis of variance (ANOVA) complemented with Tukeys test. Differences
were reported as statistically significant when p < 0.05. The statistical program used was MicrocalTM OriginTM version 8.0
(Software, Inc., US). Compared to control, there was greater inhibition of enzymatic activity in the pyloric caeca CE in the
presence of Hg2+ (55.0%), Cu2+ (54.0%), Ba2+ (46.5%) and Al3+ (28.7%) at 5 mM. In this same concentration, inhibition of
enzymatic action in the intestine CE was 53.7% (Ba2+), 51.9% (Hg2+), 44.8% (Cu2+) and 21.1% (Al3+). The Hg2+ ion at 2.5
mM inhibited 34.0% and 30.0% of trypsin activity in the pyloric caeca and intestines CE, respectively. In the presence of Cu2+
ion (2.5 mM), the inhibitions of proteolytic activities of the pyloric caeca and intestine CE were respectively 25.6% and 38.2%.
For the concentration of 0.5 mM, the highest inhibition occurred in the pyloric caeca CE using Hg2+ (20.69%) and Cu2+
(14.03%). However, there was an increase in enzyme activity in the presence of Ca2+ in the lowest concentration for both the
pyloric caeca (5.17%) and intestine (22.1%) crude extract. Except for Ca+2 ion, all the results cited were statistically different (p
< 0.05) of control activity.
Conclusions:
According to exposed, observed the negative influence of metal ions on the main digestive enzyme from E. brasilianus,
evidencing its vulnerability when exposed to these contaminants. Furthermore, these results demonstrate that this biomolecule
can serve as a biomarker of heavy metal contamination and possible recognition of anthropic impacts in the estuarine ecosystem.
Keywords: Eugerres brasilianus, Protease, Trypsin
Financial Support: CNPq, SEAPPR, FINEP/RECARCINE, FACEPE, PETROBRS AMBIENTAL and

EMBRAPA.
Resumo:31-079
INVOLVEMENT OF OXIDATIVE STRESS MARKERS IN MERCURY-INDUCED TOXICITY AND LOCOMOTOR
DEFICITS IN DROSOPHILA MELANOGASTER
Salvad, V. S. 1; Paula, M. T. D. 2; Nunes, M. E. M. 1; Rodrigues, N. R. 1; Zemolin, A. P. P. 1; Rocha, J. B.

T. D. 2; Franco, J. L. 1; Posser, T. 1
2
Departamento de Qumica,, UFSM
1
Objectives:
: Mercury (Hg) is an environmental pollutant widely used in certain types of batteries and as an essential constituent of
fluorescent light bulbs. The long atmospheric permanence of Hg0 vapor and its oxidation to the soluble inorganic form Hg(II) is
considered a cause for contamination of vast amounts of water and soil by this metal. The toxic effects elicited by Hg(II) has been
associated with accumulation of this metal in tissues or binding to endogenous thiol-containing molecules. Drosophila
melanogaster is an animal model that has been used for elucidating human diseases and in toxicological studies, and is
characterized by its easy handling and rapid reproduction cycle and development process. The aim of this study was to investigate
the effects of Hg(II) exposure on antioxidant status, Ache activity and locomotor performance in D. melanogaster.
D. melanogaster aged 5 days were exposed to HgCl2 (30-300 M) dissolved in sucrose 2M for 48 h. Finished the treatments, the
flies were used for analysis of locomotor performance (negative geotaxis), biochemical markers [activity of catalase, superoxide
dismutase (SOD)], Ache, content of non-protein thiols and lipoperoxidation induction (TBARS assay). The treatment with Hg(II)
decreased flies survival over the period of treatment. This effect was most observed after 48 hours at 100 M and 300 M
(30.12.8% and 50.44.2% decrease respectively, p
Conclusions:
Our data point the antioxidant system and the enzyme Ache as important targets of Hg(II) in D. melanogaster. It is known that an
unbalance between antioxidants and reactive molecules may trigger an oxidative cell damage which is involved in etiology of
different pathologies, including neurodegenerative diseases. In addition, this study highlights D. melanogaster as a potential
biomarker for environmental contamination with toxins implicated in human health.
Keywords: Antioxidant defenses, Drosophila melanogaster , Mercury, Oxidative stress
Financial Support: CNPq, Fapergs.
Resumo:31-080
EFFECTS OF CHRONIC TREATMENT WITH MAGNETIC IRON OXIDE NANOPARTICLES IN MALE RATS.
Wang, C. C. 2,1; Pastor, F. A. C. 1; Ruffoni, L. D. G. 1; Cardim, D. A. 1; Bogni, F. H. 1; Marangoni, V. S. 2;

Vilela, G. H. F. 2; Mascarenhas, S. 2; Zucolotto, V. 2; Nonaka, K. O. 1
1
Department of Physiological Sciences, UFSCar
2
Physical Institute of So Carlos, USP
Objectives:
The nanotechnology is a science that is suffering great development. The use of nanotechnology to develop new products to help
in diagnosis diseases, drugs and treatments for many diseases is increasing. Then, some authors have been warning against the
indiscriminate use of the nanoparticles, because their effects, toxic or not, are still unknown. Magnetic iron oxide nanoparticles
have been used in contrast liquid for magnetic resonance imaging, cancer treatment, and others. The aim of this study was to
analyze the possible effects of chronic treatment with these nanoparticles in adult rats.
(Animal Experimental Ethic Committee Protocol no. 005/2010)Wistar rats were distributed in 3 groups: Control (saline 0.9% 0.1 mL/100 of body weight, n=9), nFe 0.3 (magnetic iron oxide nanoparticles 0.3 mg/kg BW, n=8) and nFe 0.6 (magnetic iron
oxide nanoparticles 0.6 mg/kg BW, n=8). The animals were treated by gastric gavage during 8 weeks, 5 days per week. The
statistical test used to analyze the data was the t-student test (p
Conclusions:
These results showed that higher concentration of magnetic iron oxide nanoparticles used was toxic and dangerous to the rats
when take daily, but the lower concentration it was safe.
Keywords: Nanotoxicology, Nanoparticles, Magnetic Iron Oxide, Chronic treatment, Rats
Resumo:31-081
STUDY ON THE BIOCHEMICAL RESPONSES IN LAMBARIS (ASTYANAX SP), EXPOSED TO WASTEWATER
IN SANTA MARIA RIVER, ROSRIO DO SUL, BRAZIL
Cruz, L. C. D. ; Silva, F. W. D. ; Silva, D. G. D. C. ; Nunes, M. E. M. ; Salvad, V. S. ; Posser, T. ; Franco,

J. L.
Universidade Federal do Pampa, Unipampa
Objectives:
The water pollution caused by wastewater discharge from domestic sources, industrial and indiscriminate use of pesticides is a
matter of concern to modern society. It is recognized that exposure of aquatic organisms to contaminants is related to
physiological, genetic and biochemical changes, such as induction of enzymes that act in the metabolism or detoxification of
exogenous compounds. The aim of this study was to evaluate water quality in the Santa Maria River, Rosrio do Sul RS,
through the analysis of biochemical responses in Lambaris (Astyanax sp). The activity of the enzymes acetylcholinesterase,
catalase, glutathione peroxidase, glutathione reductase and glutathione S-transferase was used as markers.
The animals were collected in the Santa Maria River S1 (301459,7 S, 545445,01 O) and then transferred to aquaria in the
laboratory, where tissues were isolated (brain and muscle) and homogenized in phosphate buffer 0.1 M, pH 7. The homogenates
were centrifuged at 1000 rpm, 4 C, 5 minutes. An aliquot of the supernatant was separated for determination of
acetylcholinesterase activity. The remaining supernatant was centrifuged at 14.000 rpm, 4 C, 30 minutes. The supernatant
resulted from this second centrifugation was used for determination of catalase activity and glutathione S-transferase. The control
animals were kept in aquarium with controlled pH and temperature for a minimum period of 40 days for acclimatization. There
was a significant decrease in cholinesterase activity (78.75.8%, p
Conclusions:
In conclusion, the study points to an important degree of water contamination of water from the river Santa Maria Rosrio do Sul
- RS. More studies are needed to determine the actual levels of biological damage in the region. Moreover, the data presented in
this study demonstrate that Astyanax sp has potential for use as a bioindicator of water pollution in the region of the biome
Pampa - RS.
Keywords: Acetylcholinesterase, Astyanax sp, Catalase, Glutathione S-transferase, Pollution
Financial Support: PBDA-Unipampa, CNPq, Fapergs.
Resumo:31-082
ACUTE INTOXICATION BY CIPERMETHRIN IN JUNDIA, RHAMDIA QUELEN WITH EMPHASIS ON
CLINICAL, BIOCHEMICAL AND HAEMATOLOGICAL EFFECTS
Montanha, F. P. 3; Fredianelli, A. C. 1; Rocha, D. C. C. 1; Rocha, R. M. M. 1; Mira, R. T. 1; Galeb, L. A. G.

1
; Pimpo, C. T. 1
1
Curso de Medicina Veterinria, PUCPR
3
Curso de Medicina Veterinria, FAMED
Objectives:
Cypermethrin (CM) is a synthetic pyrethroid pesticide highly toxic to aquatic organisms. Because of its lipophilic feature it can
be highly absorbed by the fish gills, which partially explains the high sensitivity of these animals to CM exposure in
concentrations up to a thousand times lower than in mammals. This study compared clinical, biochemical and haematological
observations between CM intoxicated and non-intoxicated fish (Jundia).
The experiment involved three exposure conditions in water (0; 1.5 and 2.5 mg/L of CM) and was executed with 4 fish per
aquarium (n=36) during 96 hours. At the end of 96 hours, was performed to collect blood for hematologic and biochemical
analysis. Analyses were performed: complete blood count, plasma protein, albumin, alanine aminotransferase (ALT), aspartate
aminotransferase (AST), gamma glutamyl transferase (GGT) and alkaline phosphatase (ALP). Data were analyzed by KruskalWallis followed by Dunns test.Abnormal behavioral responses and toxic symptoms were described. Fish exposed to the CM
concentration (1.5 and 2.5 mg/L) for 96 h showed loss of equilibrium, abnormal swimming, dyspnea, keeping the operculum and
mouth open, still-vertical and sudden movements of swimming in a spiral. The fish exposed to CM (2.5 mg/L) showed
significantly higher in the values of erythrocyte (145.8 17.89cel/L), hematocrit (35.80 3.36%), hemoglobin (5.70
0.80g/dL), total leukocyte count (134.7 21.50cel/L) and AST (173.0 48.34UI/L) when compared to fish in the other groups
(122.3 18.99cel/L; 31.33 2.10%; 4.64 0.48g/dL; 104.9 28.8cel/L and 109.4 33.92UI/L, respectively). Significant
decrease in the activity levels some of the hepatic enzymes such as ALT (26.33 6.08 to 17.11 5.80 UI/L) and ALP (33.83
5.09 to 22.75 5.41 UI/L) result from CM exposure.
Conclusions:
CM caused significant biochemical changes showing a hepatic tissue in Jundia.The results show that environmental
contamination by CM can cause changes in physiology and metabolic system these animals.
Keywords: Fish, Hepatic enzymes, Pyrethroids, Sublethal concentration, Toxicity
Financial Support: PUCPR
Resumo:31-083
ACTIVITY OF ACETYLCHOLINESTERASE IN SYNAPTOSOMES CORTEX OF RATS EXPOSED TO CADMIUM
AND TREATED WITH QUERCETIN
Abdalla, F. H. ; Abdalla, F. H. ; Pereira, L. B. ; Cardoso, A. M. ; Stefanello, N. ; Pereira, R. D. ; Oliveira, L.

S. D. ; Morsch, V. M. ; Rodrigues, M. V. ; Mazanti, C. M. D. A. ; Gonalvez, J. F.
Objectives:
Considering that cadmium (Cd) is a ubiquitous environmental toxicant associated with behavioral impairments and that quercetin
possesses many therapeutic properties, this study investigated the effects of this compound on acetylcholinesterase (AChE)
activity of Cd-exposed rats.
The adult male Wistar rats were divided into eight groups (n=10-15): saline/ethanol, saline/quercetin5, saline/quercetin25,
saline/quercetin50, Cd/saline, Cd/quercetin5, Cd/quercetin25 and Cd/quercetin50. The rats received Cd (2,5mg/kg) and/or
quercetin (5, 25 or 50mg/kg) by gavage five days a week during 45 days. After the treatment the animals were submitted to
euthanasia and the cortex was removed and homogenized in 10 volumes of Medium I (pH 7.5). The synaptosomes were isolated
(Journal of Neurochemical 43; 1984) using a discontinuous Percoll gradient. The sediment was suspended in an isosmotic
solution and the final protein concentration was adjusted to 0,4 to 0,6mg/ml. AChE activity was determined (Pharmacology 7;
1961). Data were analyzed by ANOVA followed by Duncan test (p
Conclusions:
Cd inhibited the activity of AChE in synaptosomes from cerebral cortex. Quercetin at both doses reversed this inhibitory effect.
One may suggest that the inhibition caused by Cd on the activity of AChE leads to accumulation of acetylcholine, causing
overstimulation of receptors for this neurotransmitter.
Keywords: ACETYLCHOLINESTERASE, SYNAPTOSSOMES, CADIMIUM, QUERCETIN, CORTEX
Financial Support: CAPES, CNPq, FAPERGS.
Resumo:31-084
HORMONAL PROFILE OF WISTAR RATS EXPOSED TO ANTIDEPRESSANT FLUOXETINE DURING
PREGNANCY AND LACTATIONAL PERIODS
Mller, J. C. ; Zaia, R. M. ; Boareto, A. C. ; Loureno, E. L. B. ; Vechi, M. F. ; Minatovicz, B. C. ; Kienast,

M. F. ; Morais, R. N. ; Martino-andrade, A. J. ; Dalsenter, P. R.
Departamento de Farmacologia/ Universidade Federal do Paran, UFPR
Objectives:
Depression is among the most common disorders in women's health and it was estimated that nearly 10% of women will
experience depression during pregnancy and 20% after child-birth. The antidepressant treatment during pregnancy and lactation
is desirable or necessary when untreated depression increases the risk for mother, fetus or newborn. The increased use of
antidepressants in the class of Inhibitors Selective Serotonin Reuptake during pregnancy and lactation in recent years has been
accompanied by limited and often controversial information about the risk of using these drugs during these periods. That
serotonin is involved in regulating hypothalamic-pituitary-adrenal axis function has long been recognized. Thinking in this
context, this study aimed to investigate possible changes in the hormonal profile of Wistar rats exposed to the Fluoxetine
antidepressant during pregnancy and lactation.
Wistar rats ( 90 days) were exposed to Fluoxetine (0.4, 1.7 and 17 mg/kg p.o.) or distilled water (5 ml/kg p.o.) from day 7 of
pregnancy to day 21 of lactation. Individual 24-hours faeces samples were collected of the dams during pregnancy (days 1, 7, 14
and 21) and during lactation (days 7, 14 and 21) of all treatment groups (9 to 15 female pregnant per group). Identified samples,
stored at -20 C, were weighed and submitted to extraction and determination of levels of fecal metabolites of progesterone,
estrogen and corticosterone (MPF, MEF and CMF, respectively) by enzyme immunoassay (Ann. N.Y. Acad. Sci. 1046, 5474,
2005). The experimental protocol was approved by the Ethics Committee for Animal Experimentation under N 347. Data were
analyzed by KruskalWallis followed by Dunns test. The results obtained showed that on the 15th day of pregnancy a significant
increase of 255% in the levels of metabolites of progesterone and 141% in metabolites of corticosterone in rats exposed to 17
mg/kg of Fluoxetine when compared with the control group. Hormonal change was also observed on day 7 of lactation, which
revealed a significant increase of 211% in the metabolites of progesterone and 137% in the metabolites of estrogen in rats
exposed to the same dose of Fluoxetine compared with control rats. Many studies have shown that serotonin increases the
secretion of adrenocorticotropic hormone (ACTH), and that Fluoxetine increases secretion of ACTH by an increase in the
hypothalamic secretion of both corticotropin-releasing factor and arginine vasopressin, beyond can act directly on the adrenal
gland. The increase of fecal hormone metabolites observed may have been caused in part by increased serotonergic stimulation
that Fluoxetine exerted on the release of ACTH, causing an increased synthesis of hormones by the adrenal cortex.
Conclusions:
We conclude that exposure of Wistar rats during pregnancy and lactation to antidepressant Fluoxetine at a dose of 17 mg/kg
caused changes in hormonal profile of them. Further studies are needed to investigate if these hormonal changes caused by
Fluoxetine could harm embryonic and fetal development.
Keywords: depression, fluoxetine, hormonal profile, lactation, pregnancy
Resumo:31-085
EVALUATION OF THE REPEATED DOSE TOXICITY OF PERSEA AMERICANA LEAVES EXTRACT IN RATS
Lima, C. R. 1; Vasconcelos, C. F. B. 1; Limeira, M. M. F. 1; Costa-silva, J. H. 2; Wanderley, A. G. 1,3

1
Depto Cincias Farmacuticas, UFPE
2
Depto Educao Fsica e Cincias do Esporte, UFPE
3
Depto Fisiologia e Farmacologia, UFPE
Objectives:
Persea americana Mill. (Lauraceae) has been used to treat hypertension, dyslipidemia and diabetes. Although its wide use, no
toxicological studies were performed. Thus, we evaluated the toxicity of hydroalcoholic extract of Persea americana leaves on
biochemical and hematological parameters of rats.

Male Wistar rats (aged 3 months, weighing 260-280 g) were randomly divided into three groups (n = 10/group). Animals
received orally water (control group, C) or the hydroalcoholic extract of Persea americana leaves at the doses of 0.15 (T1) and
1.5 g/kg/day (T2) for 30 days. At the end of the treatment, animals were anesthetized with thiopental (0.035 g/kg, i.p.) and the
blood samples were collected for biochemical and hematological analysis. The results were compared by one-way ANOVA
followed by Newman-Keuls test. No toxic signs or deaths were observed, similarly no significant differences were observed in
body mass gain, food and water consumption following whole treatment. In biochemical analysis, it was observed an statistical
difference in alanine aminotransferase values (C = 50.21.9, T1 = 58.42.8* and T2 = 66.92.1* U/L), but no differences in
glucose (C = 92.96.1, T1 = 92.74.6 and T2 = 87.34.3 mg/dL); blood urea nitrogen (C = 33.41.2, T1 = 31.34.8 and T2 =
29.24.0 mg/dL); uric acid (C = 1.030.06, T1 = 1.220.06 and T2 = 0.990.09 mg/dL); creatinina (C = 0.710.03, T1 =
0.730.03 and T2 = 0.690.01 mg/dL); aspartate aminotransferase (C = 108.25.9, T1 = 101.85.1 and T2 = 124.910.2 U/L);
gamma-glutamyl transpeptidase (C = 6.00.4, T1 = 6.10.2 and T2 = 6.00.2 U/L); total cholesterol (C = 99.611.1, T1 =
89.94.9 and T2 = 81.53.7 mg/dL); alkaline phosphatase (C = 138.912.7, T1 = 130.112.4 and T2 = 147.46.3 U/L); amylase
(C = 498.233.2, T1 = 464.933.7 and T2 = 541.241.0 U/L); total bilirubin (C = 0.1000.005, T1 = 0.1100.007 and T2 =
0.1000.004 mg/dL); direct bilirubin (C = 0.070.01, T1 = 0.070.006 and T2 = 0.060.008 mg/dL). Likewise, no changes were
observed in hematological parameters: erythrocytes count (C = 8.40.1, T1 = 8.30.6 and T2 = 8.20.3 106/uL); hemoglobin (C
= 16.10.3, T1 = 15.81.0 and T2 = 15.40.5 g/dL), hematocrit (C = 47.20.9, T1 = 45.52.6 and T2 = 45.21.7%); MCV (C =
56.01.2, T1 = 55.71.6 and T2 = 55.40.3 um3); MCH (C = 19.10.2, T1 = 19.20.4 and T2 = 18.90.3 pg); MCHC (C =
34.10.6, T1 = 34.60.3 and T2 = 34.30.5 g/dL); RDW (C = 13.10.6, T1 = 14.50.7 and T2 = 13.50.3 %); platelets count (C
= 750.428.7, T1 = 791.223.4 and T2 = 637.556.0 103/uL); white blood cell (C = 16.01.0, T1 = 16.31.2 and T2 = 14.51.3
103/uL); neutrophils (C = 10.40.9, T1 = 10.31.1 and T2 = 13.40.9 %); eosinophils (C = 0.90.3, T1 = 0.50.3 and T2 =
0.80.4 %) and lymphocytes (C = 84.91.1, T1 = 84.42.0 and T2 = 80.81.1 %). Although there was statistical difference in
alanine aminotransferase among groups, the values remained under the reference range for the species.
Conclusions:
Oral treatment with hydroalcoholic extract of Persea americana leaves does not produce any toxic effect on the biochemical and
hematological parameters of male Wistar rats.
Keywords: Lauraceae, Persea americana Mill., Repeated dose toxicity, Biochemical and hematological parameters
Financial Support: UFPE
Resumo:31-086
PULMONAR EFFECTS INDUCED BY TITYUS OBSCURUS SCORPION THE VENOM IN RATS
Silva, A. P. S1,2; Candido, D. M. 2; Abraho, A. L. N. 1; Dorce, V. A. C. 1

1
Farmacologia/ Instituto Butantan, IBu
2
Artropdes/Instituto Butantan, IBu
Objectives:
Scorpions of medical importance in the Brazilian Amazon belong to the Tityus genus and Tityus obscurus is the responsible for
the greater number of accidents in the Northern area of the country. This work aimed to verify if Tityus obscurus scorpion venom
induces pulmonary oedema as observed to others Tityus species.
The scorpions were captured in the west region of Par State and submitted to electrical stimulation for the venom obtainment.
The dose of 10mg/kg diluted in NaCl 1.46% was used. Male Wistar rats (230-260 g) were separated in 3 experimental groups
injected with scorpion venom and euthanatized 1, 4 or 6 hours after injection, and 1 control group injected with NaCl 1.46%
(1ml/kg) and euthanatized 1 hour later. It was utilized 5 animals per group injected intraperitoneally. The lungs of animals were
removed free from trachea and weighed. The severity of the pulmonary oedema was estimated through the lung/body index (lung
weight X 100/ body weight). Student t-test was used for statistical analyze, p
Conclusions:
The Tityus obscurus scorpion venom evoked some pulmonary effects without pulmonary oedema. This kind of effects is different
from that one induced by the venoms of others Tityus that cause pulmonary oedema with smaller doses and less intense
hemorrhagy.
Keywords: efeito pulmonar, veneno, Tityus obscurus
Financial Support: INCTTox, CAPES
Resumo:31-087
DOES URBAN PARTICULATE MATTER ACT DIRECTLY IN THE CENTRAL NERVOUS SYSTEM OF RATS?
Fagundes, L. S. 1; Rhoden, C. R. 1; Gamaro, G. D. 1; Saldiva, P. H. N. 2; Zanchi, A. C. T. 1; Morais, M. C. 1;

Fleck, A. D. S. 1
1
Ps graduao / UFCSPA, UFCSPA
2
Universidade Federal de So Paulo, USP
Objectives:
Particulate Matter (PM) is the major air pollutant in urban areas and it has been implicated as a source for serious health problem.
PM toxicity against respiratory tract, by oxidative stress and inflammation pathways, is well-known. Recently, some researches
demonstrate PM neurotoxicity in animal models and human. The aim of the present study was to evaluate, in vitro, the different
responses in terms of oxidative stress induced by PM incubation in the cortex, cerebellum, striatum, hippocampus and olfactory
bulb of rats developed by PM oxidative damage. In addition, to determine whether alpha-tocopherol, a wellknown non-enzymatic
antioxidant, may prevent the oxidative stress induced by urban particles.
Olfactory bulb, cerebral cortex, cerebellum, striatum and hippocampus of Wistar adult male rats (n=32) were homogenized to
perform the incubations and oxidative stress measurements. The in vitro preventive effect of alpha-tocopherol (40M) preincubation and oxidative action of PM (3g/100L) incubation were evaluated by thiobarbituric acid-reactive substances test
(TBA-RS), antioxidant enzymatic activity of catalase (CAT) and superoxide dismutase (SOD). Statistical Analysis were
performed by one-way analysis of variance (ANOVA) followed by the Post hoc multiple comparisons among Student Newman
Keuls test. The level of significance was set at 5%. Lipid peroxidation measured by TBA-RS was significantly enhanced in
hippocampus exposed to PM which was not prevented by alpha-tocopherol treatment (p=0.002). Pre-incubation with alphatocopherol isolated reduced TBA-RS levels in striatum (p=0.005) and in cerebellum (p=0.018). SOD was greatly lower in
hippocampus (p=0.002) and striatum (p=0.001) exposed alpha-tocopherol pre-incubation. Besides, CAT activity was decreased in
olfactory bulb exposed to -tocopherol plus PM or only to -tocopherol (p=0.012).
Conclusions:
These results suggest that hippocampus is the most affected brain structure by PM direct exposure and it may occurs by oxidative
stress pathway, besides alpha tocopherol (40M) pre-incubation did not prevent oxidative response.
Keywords: CENTRAL NERVOUS SYSTEM, RATS, PARTICULATE
Financial Support: FAPERGS and Conselho Nacional de Desenvolvimento Cientfico e Tecnolgico

CNPq
Resumo:31-088
EVALUATION OF TOXIC EFFECTS IN MOTOR COORDINATION, SPONTANEOUS LOCOMOTOR ACTIVITY
AND TEMPERATURE OF MICE CAUSED BY THE ASSOCIATION OF P-SINEPHRINE, EPHEDRINE, SALICIN
AND CAFFEINE: A MIXTURE CONTAINED IN WEIGHT LOSS PRODUCTS
Schuh, R. S. 1; Jacques, A. L. B. 1; Schmitt, G. C. 1; Arbo, M. D. 1; Leal, M. B. 1; Dallegrave, E. 3,4;

Limberger, R. P. 1
3
Centro de Informao Toxicolgica , CIT-RS
1
Laboratrio de Toxicologia da Faculdade de Farmcia, UFRGS
2
Laboratrio de Farmacologia do ICBS, UFRGS
4
Fundao Estadual de Produo e Pesquisa em Sade , FEPPS
Objectives:
Dietary supplements and weight loss products containing the association of p-synephrine, ephedrine, caffeine and salicin are
largely used, even if ephedrine is prohibited in many countries. However, the safety of this association is still unknown due to
lack of studies on the toxic effects of it, coupled with the growing number of reports of adverse reactions - as cardiovascular
problems - and even deaths related to the use of these products. The aim of this study was to evaluate the effects of p-synephrine
in association with ephedrine, salicin and caffeine through the spontaneous locomotor activity test, the locomotor coordination
test and body temperature of mice.
To the spontaneous locomotor activity test, the CF1 male mice were individually habituated in activity cages for 10 min and then
received the treatments (n=7-10) by oral gavage: water (control) or a 300, 350 or 400 mg/kg aqueous mixture of p-synephrine,
ephedrine, salicin and caffeine (10:4:6:80 w/w, respectively). After 30 minutes the animals returned to the activity cages and the
number of crossings was recorded for 15 min. The test showed that the number of crossings in the group treated with the mixture
of p-synephrine, ephedrine, salicin and caffeine at 300 mg/kg (68.313.3), 350 mg/kg (66.49.1), and 400 mg/kg (59.410.8)
was significantly reduced (pp-synephrine, ephedrine, salicin and caffeine aqueous mixture 30 minutes after the administration. It
was observed a significant reduction in body temperature of mice (p
Conclusions:
These findings indicate a depressant effect of the mixture and an increase in toxicity. All the substances used in this test have a
direct or indirect adrenergic stimulation and the association of them could potentiate the stimuli and enhance toxicity. So,
products containing the questioned association should be used with caution until the toxicological profile is totally elucidated.
Keywords: caffeine, ephedrine, p-synephrine, salicin, weight loss products
Financial Support: CNPq/Brazil
Resumo:31-089
ACUTE TOXICITY OF SUBSTANCES COMMONLY USED IN WEIGHT LOSS COMPOUNDS AND DIETARY
SUPPLEMENTS
Jacques, A. L. B. 1; Schuh, R. S. 1; Schmitt, G. C. 1; Arbo, M. D. 1; Dallegrave, E. 2; Leal, M. B. 3;

Limberger, R. P. 1
1
Faculdade de Farmcia, Dep. de Anlises, Lab. de Toxicologia, UFRGS
2
Centro de Informao Toxicolgica, CIT
3
Instituto de Cincias Bsicas da Sade, Lab. de Farmacologia, UFRGS
Objectives:
The use of natural products in weight loss compounds and dietary supplements lead to more attention on their toxicological
potential. The security and effectiveness of products containing an association of p-synephrine (from Citrus aurantium L.),
ephedrine (from Ephedra sinica Stapf), salicin (from Salix sp.) and caffeine (from Paullinia cupana Kunth, Cola nitida Vent
Schott&Endl.) are controversial and there are many reports of cardiovascular problems related to the use of this mixture. Thus,
the aim of this study was to investigate the acute toxicity of p-synephrine associated with ephedrine, salicin, and caffeine in mice.
The acute toxicity was performed according to OECD Series on Testing and Assessment Number 24. Male and female (n=6 per
sex per group) albino CF1 mice, weighing 40.1 5.2g and 33.1 2.0g respectively, were treated by oral gavage with a 300, 350
or 400 mg/kg aqueous mixture of p-synephrine, ephedrine, salicin and caffeine (10:4:6:80 w/w, respectively) or water (control).
The animals were observed for one minute at 5, 15, 30, 60, 120, 240, 300, 360 minutes and 24h after administration. The specific
signs monitored were alterations in the locomotor activity, stimuli reaction, piloerection, salivation, gasping, tremors and
seizures. Lethality was daily monitored for 14 days and the surviving animals were euthanized and necropsied. Macroscopic
alterations in vital organs such as heart, liver, spleen, lungs, kidneys and adrenal glands were analyzed (University Ethics
Comittee number 2007982). The LD50 was calculated using Probit analysis (Software IBM SPSS statistics 17).All doses tested
in the current work, for both male and female animals, led to episodes of ptose, piloerection, and alterations in locomotor activity.
At first moment, a locomotor activity reduction was observed in animals treated with 300mg/kg of the mixture. An increase of
this sign occurred after 120 minutes. Jumping episodes were also observed at the same dose. Animals showed the same signs
including agitation between 120 and 240 minutes at 350mg/kg. Tearing, gasping, tremors and muscle spasms were also presented
at 400 mg/kg. Higher doses led to deaths and seizures, only observed in male, one at 350 mg/kg and five animals at 400 mg/kg.
The necropsy showed pulmonary hemorrhage. Female mice presented the same signs of toxicity than male, but they were less
intense, since no jumping episodes neither death occurred. No other macroscopic changes and weight alterations were observed
in the organs. The estimated LD50 was 374,1 mg/kg for male animals. Since no deaths were observed in females, the LD50
wasnt determinate for this group.
Conclusions:
The intensity of the effects increased directly with the dose. Differences between male and female effects were observed, being
more intense in males. The LD50 of the mixture of p-synephrine, ephedrine, salicin and caffeine was estimated in 374,1 mg/kg
for male animals. Finally, the commonly used association demonstrated acute toxicity. Even more studies should be done to
establish the whole toxicological profile of the association and the differences observed between females and males.
Keywords: p-synephrine, ephedrine, salicin, caffeine, acute toxicity
Financial Support: Conselho Nacional de Desenvolvimento Cientfico - CNPq
Resumo:31-090
HEPATIC EFFECTS OF FXR AGONIST IN ALCOHOLIC HEPATIC STEATOSIS MODEL.
Lvero, F. R. ; Dreifuss, A. ; Bastos-pereira, A. L. ; Stolf, A. M. ; Chocorski, R. ; Acco, A.

Universidade Federal do Paran, UFPR
Objectives:
Alcoholic liver diseases are among the most important consequences of excessive or long-lasting ethanol use, and represent one
of the major causes of morbidity and mortality worldwide. Intracellular accumulation of lipids (steatosis) is the most common
injury in heavy drinkers, and although regarded a benign process, there is evidence that alcoholic fatty liver disease contributes
significantly to the progression of liver damage. Ethanol metabolism generates toxic molecules and reactive oxygen species
(ROS) resulting in oxidative stress, initiation of cellular injury and apoptosis, thus contributing to the progression of liver lesions.
The modulation of oxidative mechanisms in these scenarios, however, is yet to be fully elucidated. Likewise, there is still no
research on the involvement of Farnesoid X Receptor (FXR), a nuclear receptor involved in lipid and glucose metabolism, in
these pathological processes. The purpose of this study was to evaluate the co-participation of both oxidative stress and FXR in
the pathogenesis of alcoholic fatty liver disease.
All the protocols were approved by institutional ethic committee for animal research (CEEA) and received certificate n. 438.
Swiss male mice (8-10 weeks) were separated in 2 groups (n = 12), which received liquid diet containing 10% ethanol or water
(control group) for 6 weeks, as well as a low protein diet (6%). In the last 15 days of the diet, mice that had received ethanol or
water were separated again for oral treatment, performing 4 groups in the total. From these groups, 2 received FXR agonist
6ECDCA (1 mg/Kg dissolved in Tween 1%) and 2 received Tween 1% (vehicle). Following this treatment, the animals were
anesthetized with ketamine and xylazine for blood and liver samples collection, in order to perform serum biochemical assays
(AST, ALT), and measurement of hepatic oxidative stress parameters (Superoxide Dismutase (SOD), Gluthathione (GSH), Lipid
Peroxidation (LPO) and Gluthatione-S-Transferase (GST)). These parameters were expressed as the amount of protein found in
each liver sample using the Bradford method. All results were statistically analyzed by ANOVA followed by Bonferroni test as
post hoc (p
Conclusions:
The presented data show that ethanol consumption associated with hypoproteic diet, a model for alcoholic liver steatosis, can
reduce both GSH level and SOD activity, indicating an alteration in hepatic redox balance. The FXR agonist 6ECDCA seems to
protect the liver against these alterations. Further experiments regarding this compound should be encouraged, because a possible
therapeutic action of FXR agonists in alcoholic fatty liver disease can be speculated.
Keywords: Alcoholic Hepatic Steatosis, Farnesoid X Receptor (FXR), FXR agonist
Financial Support: Fundao Araucria and REUNI.
Resumo:31-091
TOXICOPATHOLOGY OF SIMVASTATIN INDUCED HEPATIC DAMAGE: A MODEL FOR NASH?
Renn, A. L. ; Souza, P. C. ; Barbosa-souza, V. ; Freitas, C. P. ; Schenka, A. A.

Farmacologia- Universidade Estadual de Campinas, UNICAMP
Objectives:
Nonalcoholic Steatohepatitis (NASH) is a silent syndrome with increasing incidence among obese, diabetic and metabolic
syndrome patients. NASH can progress to liver fibrosis ( in 27% of the cases) and cirrhosis (in 19% of the cases), and is
considered an important cause of cryptogenic cirrhosis. Currently, treatment of NASH is based solely on control of underlying
diseases (when they can be identified), with no specific measures directed against the liver lesions. During toxicopathologic
studies on statins in murine models performed in our laboratory, we observed the occurrence of histopathological signs of NASH
in animals using simvastatin. These findings led us to study the microscopic hepatic lesions associated with oral administration of
simvastatin with the aim of characterizing a new pharmacological model of NASH in experimental animals
Sprague-Dawley rats (~200g) were treated with simvastatin (purity>95%) orally for 7 consecutive days at doses 20, 40, 60, 80
and 100 mg/kg/day (n=6 each group). Simultaneously, a control group was composed by animals receiving only drug vehicle (in
this case soy oil). After euthanasia, the livers were removed, histologically processed and analyzed for the presence of the main
morphological features that allow for the diagnosis and staging of NASH according to the criteria of the Brazilian Society of
Pathology (including steatosis,ballooning anf lobular inflammation) . We also determined the plasma levels of liver enzymes
(ALT and AST) by colorimetric method. The experimental protocols were approved by an instructional Committee for Ethics in
Animal Experimentation (CEEA/UNICAMP, protocol n. 2335-1). In the present protocol, 90% of animals treated with
simvastatin developed laboratory features compatible with NASH, and/or at least borderline histological features of this disease.
Animals treated with 100 mg/kg/day significantly increased serum levels of ALT (112.8929.24 vs 58.782,08 U/mL of the
control group) and AST (108,8152,39 vs 53,9925,82 U/mL of the control group) (p
Conclusions:
These results indicate that the oral administration of simvastatin (specially in doses greater than 80mg/kg/7 days) is a promising
model of induction of NASH, which may be useful in future studies for the development of new pharmacological strategies for
the specific treatment of NASH in humans.
Keywords: NASH, Simvastatin, Hepatic Damage, Toxicolpathology, Model
Financial Support: CAPES and FAPESP

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XXVI Reunio Anual da FeSBE - FeSBE 2011

Fernandes, T. 1; Hashimoto, N. Y. 1; Magalhes, F. C. 1; Fernandes, F. B. 3; Casarini, D. E. 3; Carmona, A.

Financial Support: FAPESP, CAPES and CNPq

THE ROLE OF TYPE 2 ANGIOTENSIN II RECEPTOR (AT2) IN THE CARDIOPROTECTIVE RESPONSE

Financial Support: FAPESP, CNPq.

Gonalves, M. C. ; Sordi, R. ; Assreuy, J.

Financial Support: CNPq, CAPES and FAPESC.

Mendes, L. V. P. 1; Dias,m. S. 1; Boldrini, L. C1; Takiya, C. M. 1; Oliveira-pinto, L. M 1; Nascimento, J. H.

Methods and Results:

Financial Support: Projeto Casadinho-CNPq; PROCAD-CAPES; FAPERJ Primeiros Projetos, Programa

Dias, L. O. 2,1; Durand, M. T. 1; Castania, J. A. 1; Fazan Jr. , R. 1; Salgado, H. C. 1

Financial Support: Capes, CNPq, FAPESP

Ferraz, E. F. ; Oliveira, A. P. F. ; Barros, F. ; Riva, D. R. ; Nascimento, J. H. M.

Financial Support: FAPERJ, CNPq

Lacerda-miranda, G. ; Vieira, A. K. G. ; Sores, V. M. ; Bernardes, A. F. ; Lessa, J. G. ; Cunha, A. C. S. R. ;

Financial Support: CNPq , FAPERJ, Vital Brasil

Lino, C. A. ; Barreto - Chaves, M. L. M.

Financial Support: CAPES

Farah, H. M. A. T. 1; Almeida, R. L. 1; Mostarda, C. T. 2; Colombari, D. S. A. 4; Farah, V. M. A. 3; Sato, M.

Financial Support: PIBIC-CNPq, FAPESP and NEPAS.

Freire Jr, D1; dos Santos, L. 1; Fiorim, J. 1; Stefanon, I. 1; Vassallo, D. V. 1,2

Financial Support: CNPq,CAPES, FAPES

Almeida, S. A. 1; Tiradentes, R. V. 1; Borgo, M. V. 1; Santuzzi, C. H. 1; Moraes, A. N. 1,2; Claudio, E. R. G.

Methods and Results:

Financial Support: CAPES and CNPq

Tiradentes, R. V. ; Santuzzi, C. H. ; Borgo, M. V. ; Claudio, E. R. G. ; Brum, M. F. ; Endlich, P. W. ;

Financial Support: CAPES and CNPq (Brazil)

Hamada, R. Y. ; Cunha, N. V. D. ; Reis, D. A. S. ; Pinge, M. C. M. ; Gerardin, D. C. C. ; Gomes, G. G. P.

Financial Support: PIBICUEL

Matsumoto, A. K. ; Silva, A. M. D. ; Abreu, S. B. D. ; Filhho, P. P. ; Pinge, M. C. M.

Financial Support: PIBIC/UEL.

Penitente, A. 1; Novaes, R. D. 1; Natali, A. J. 1; Silva, M. F. D. 1; Ribeiro, M. F. 3; Fernandes, K. M. 1;

Financial Support: FAPEMIG, CNPq, CAPES, FUNARBE, UFV

Urzedo-rodrigues, L. S 1; Ferreira, H. S. 2; Almeida, D. O1; Batista, A. S. 1; Dias, D. P. M. 3; Fregoneze, J.

Financial Support: Capes, CNPq, FAPESB

Arantes, P. C. ; Rodrigues Jr, L. F. ; Medei, E. ; Nascimento, J. H. M.

Methods and Results:

Financial Support: FAPERJ and CNPq/PIBIC

Souza, P. R. M. D. 1; Moreira, E. D. 1,1; Mostarda, C. 1; Monteiro-de-moraes, W. M. A. 2; Guimares, F. 2;

Financial Support: CAPES

Manrique, N. ; Pereira, C. C. S. ; Okamoto, R. ; Antoniali, C.

Financial Support: Fapesp (2008/01893)

Melo, S. F. S. ; Junior, M. A. C. ; Bozi, L. ; Drummond, L. R. ; Natali, A. J. ; Oliveira, E. M. D.

Financial Support: FAPESP

Munhoz, F. C. 1; Potje, S. R. 1; Hildebrand, M. C. 1; Pereira, A. C. 2; Ramos, L. C. B. 2; Bendhack, L. M. 2;

Financial Support: CNPQ

Barbosa, R. M. ; Barbosa, S. P. ; Freiria-oliveira, A. H. ; Menani, J. V. ; Colombari, E. ; Colombari, D. S.

Financial Support: PIBIC-CNPq, FAPESP

Almeida, J. F. Q. ; Sobrinho, D. B. S. ; Alves, G. M. M. ; Dourado, J. G. ; Macedo, L. M. ; Mendes, E. P. ;

Financial Support: CNPq/INCT-Nanobiofar.

de Pr, M. A. ; Poli, A. ; Linder, A. E.

Keywords: SEROTONIN, UPTAKE, VASCULAR REACTIVITY, VEIN

Financial Support: CNPq, FAPESC/CNPq (005/2009), Funpesquisa.

Silva, J. S. D. 1; Zapata-sudo, G. 1; Seabra, S. 4; Sousa, P. J. D. C. 2; Resende, A. C. 3; Moura, R. S. 3; Sudo,

Financial Support: FAPERJ, Capes, CNPq, INCT/INOFAR

Potje, S. R. 1; Munhoz, F. C. 1; Hildebrand, M. C. 1; Ramos, L. C. B. 2; Bendhack, L. M. 2; Antoniali, C. 1

Financial Support: CNPq, FAPESP

Rossi, M. V. ; Schoorlemmer, G. H. ; Cravo, S. L.

Simes1; Batista1; Fiorim1; Lima1; Stefanon1; Vassallo1,2

Financial Support: CAPES / CNPq / FAPES- FUNCITEC

Simes, M. R. 1; Furieri, L. B. 1; Fioresi, M. 1,2; Broseghini, G. B. 1; Vescovi, M. V. A. 3; Faria, T. de O. 1;

Financial Support: CAPES, CNPq e FAPES

Pavan, C. G. ; Penasso, M. G. ; Barbosa, S. P. ; de Luca Jr. , L. A. ; Colombari, D. S. A. ; de Paula, P. M. ;