Tips

Chapter 12- biochemistry

Carbohydrate biochemistry
Catabolism- breaking down of molecules- it is anerobic and requires energy- examples are
glycoylsis when glucose or pyruvate from glycolysis is converted to carbon dioxide and water
Annabolism- formation of molecules- it doesnt require energy for example formation of the
porphyrin ring in heme-
Fermentation is a reaction in which ethanol or lactate are fromed from glucose. It is anaerobic
Carboydrate metabolism include- glycogenesis, glycogenolysis, gylcolysis, glucogenesis.
Glycogenesis- formation of glycogen
Glycolysis- break down of glucose
Glucogenesois- formation of glucose
Glycogenolysis break down of glycogen
In anaerobic conditions glycolysis occurs in cells without mitochondria- and glucose is converted
to lactic acid
Citric acid cycle is the krebs cycle and is break down or synthetic purposes occurs in the
mitochondria
In aerobic conditions = pyruvate from glycolysis enters the krebs cycle and then it is oxidized to
water and carbon dioxide if there is not enough 02 pyruvate is converted to lactate
Protein biochemistry
Amino acids are building blocks of proteins.
Proteins are formed from a polymerization reaction between two amino acids and cause a
bonding (peptide bond)
Essential amino acids are those that can not be synthesized in the body and need to be
gotten from the diet- (PVT TIMHALL)
Essential Aminoacds
Phenylalanine
Valine
Trypthophan
Theorine
Isoleucine
Methionine
Histidine (infants)
Argenine (infants)
Leucine
Lysine

Amino acid derivatives

PhenylalanineTyrosine dopadopamine norepherine ephedrine
Enzymes-> Tyrosine hydroxylase dopa decarboxylasedopamine b hydorxylase
phenlethanolamine n methyl transferase

Dopamelanin

Tryptophan 5 hydroxytryoptopham 5 hydroxy triptamine

Tryptophan niacin

Tryptophan melanonin

Glycine porphyrin ring heme

Arginine NO vasodilator

Arginine urea

Arginine creatinine

Glutamate GABA

Amino acids- acidic side chain- aspartic acid and glutamic acid
Amino Acids basic side chain- argine. Lysine, histadine
Amino Acids- Polar and non ionic- Glycine, serine cystine
Amino Acids non polar and hydrophobic- Alanine, Valine, Leucine, Methionine,
Trypthophan, Phenylamine, Leucine, Isoleucin

Amino acids are zwitterions- this means that they have no net charge as they are made up of +ve
and Ve ions. 50% is dissociated and they have 2 titration curves.
Isoelectric point (pl) is the point in which there is no net charge
pH>pl (-ve charge)
PH<pl (+ve charge)
There are four structures-
primary structure: sequence of amino acids- location of disulfide bonds
Secondary structure- arrangement of proteins in an alpha helix or beta sheath
Tertirary- 3d structure
Quatanery- arrangement of individual subunit chains into a complex molecue

Simple proteins- are naturally occurring proteins and hydrolysis yields only alpha amino acds
eg albumin, prolamine glutelin
Conjugated proteins- classified on the nature of their prosthetic groups-
Derived proteins- are formed from primary or conjucated proteins by the actions of an acid-
alkali- heat water-alcohol or an enzyme. They differ in chemical and physical properties of
the primary or secondary derived protein.

Protein Denaturation
When the structure of the protein is unfolded and the structure gets disorganized due to the
base pairs being broken. Proteins not denatured by hydrolysis but denatured by heat, ph,
alcohol or enzymes. The process is usually irreversaible

Globular and Fibrous proteins
Globular proteins- Heme protens are specialized proteins- and they contain tightly bound
prosthetic groups- Haemoglobin and myoglobin are the most common heme proteins in
humans. They bind to oxygen reversaibly and they have a high affinity to carbon monoxide
Haemoglobin- is a complex ring- of porphyrin and of ferrous ion (fe2+) It transports oxygen
in the blood- and CO and CO2 bind reversailby with haemoglobin
Myoglobin- is a complex of prorpyrin ring and Fe2+- transports oxygen in the tissues and the
muscles and skeletal muscles
Hemoglobinopathies- eg for thalsemia and for sickle cell anemia
4 pyrol rings make this porphyrin ring in Haemoblobin and
myoglobin
Methamoglobin (fe3+) can not bind to oxygen and is converted to haemoglobin by
methamoglobin reductase.

Fats and lipids
In fatty acid synthesis- the end product is Palmitate
It is associated with Hexose monophosphate shunt,

Lipids are divded into 5 classes due to their structure

Glycolipids
-known as cerbrosides
-Isolated from the brain
-in hydrolysis they yield fatty acid, galactose and sphingosine
-Known as galactolipids due to presence of galactose- such as phrenosin and kerasin
-
Sphingolipids- formed from palmitoyl CoA and serine- Sphingosine forms a ceramic backbone
when joined to fatty acids-
Sphingosine + (sugars, choline phosphate, sialic acid)= cerborsides or shingomyelin found in
nerve tissues and membrane.

Phospholipids- known as phosphatides- they are esters that consist of fatty acid and phosphoric
acid eg lethicin- form part of membranes

Sterols (steroids)
the sterols are alcohols and structurally related to steroids
They have 3 cyclohexane ring and 1 cyclopentane ring
The steroids are converted to compounds such as bile, vitamin D and are not broken down
completely


Waxes
High molecular weight esters
Made up of monohydric alcohol and fatty acids eg spermaceti

Fixed oils and fats- Esters of glycerol and fatty acids
fixed oil solid at room temperature is called fat

Essential Fatty acids
-Linoleic acid, arachiondinc acid, and these are broken down by acetyl co a and enter the krebs
cycle- very rapid break down causes ketoacidosis in diabetes

Fatty acid synthesis is cytosolic and the end product is palamitic acid. Acetly co A is the
substrate-

Cholesterol synthesis- the RLS is 3 hydroxy- 3 methylglutaryl co A reductase
Acetyl co A 3 hydoxy 3 melthyl glutraly co A
HMG Co A - melvanonate
melvanoate isoprene
6 xIsoprene squalene
Squalene lanosterol
Lanosterol cholesterol


Essential trace elements
Copper- Deficiency Wilkinsons disease
leucopenia, neutropenia, mimic (cu2+) defect
present in cytochrome oxidase

Iron
-most abundant metal in the body
-microcytic/hyopchonic anemia is deficiency
-Hemochromatosis- in high doses

Zinc
Children- poor growth
Adults sexual defiecincy

Selinium- Deficincy causes cardiomyopathy

Chromium- impaired glucose tolerance

Molybendum- present in xanthine oxidase needed to convert purine to uric acid

Iodine- deficiency can cause goiter disease (diffuse toxic goiture , toxic non glodular goiter)


TIPS

Nitric oxide is derived from arginine -
Arginine also gives creatinine and urea

Serotonin is produced by tryptophan
Tryptohan- 5-hydoxy trypophan5 hydroxy triptamine (serotonin)

Phenylketanuria- is excessive phenylalanine in the urine

Carbon monoxide has affinity Haemoglobin, myoglobin and cytochrome oxidase

Carboxyhaemoglobin- Carbon monoxide and oxy haemoglyobin

Iron in haemoglobin is in the ferrous state

Rate limiting step in cholesterol synthesis is HMG CO A reductase

Essential fatty acids- lineloic and linelonic (omega 3 and omega 6)

Eiconosaids are prostaglanids

Basic amino acid is arginine also is histamine and lycine

Amino acid with a negative and positive charge- zwitter ion

They have 2 tituration curves

Ph>pl - -ve charge

Ph <pl- positive charge

Energy stored in the body in glycogen

Starch made up of glucose

Leithicins are phospholipids

Sphingolipids contain sphinogsne and are glycolipids

End product of anaerobic glycoyisi- lactate lactic acid

End product of aerobic glycosis is pyruvic acid

Krebs cycle in the mitochondria

Gylcolysis occurs in the cycoplasm

End product or purine is uric acid

Interstitial fluid surrounds the cells so the ion that is most extracellular is sodium, calcium and
chloride- if inside the cell most abundant is potassium, magnesium and phosphate



Lineloic acid (omega 6-) above is lineloic acid as the c=c is on 6 carbon atom!
Linolic acid (omega 3)




13- Clinical Biochemistry
Renal Function test- Crcl and BUN(blood urea nitrogen)
Liver function test- LDH, AST, ALT, ALP
Cardiac enzymes- TROPIONINS, CREATINE KINASE
Urine analysis- KETONE BODIES, PH , SPECIFIC GRAVITY
Blood works- CBC,
Anticoagulants- INR- WARFARIN (2-3 INR), HEPARIN (APTT), LMWH
Thyroid test: SERUM TSH (0.5-5MU/L) TT4, FT4, TT3, FT3
Blood urea nitrogen is an end product of protein metabolism- it is produced In the liver and
excreted by the kidneys- in normal conditions the urea clearance is 60% GFR. BUN increase
indicates renal disease.
Normal values for BUN- 8mg/dL to 18mg/dL (3 to 6.5mmol/L) the concentration of the BUN
reflects renal function- because the urea nitrogen in the blood is filtered completely at the
glomeruli of the kidney. Then it is absorbed and tubuar secreted within the nephron.
Increase in BUN indicated acute renal failure. The BUN decrease may indicate a terminal liver
disease- as the liver only produces BUN. Increase in BUN is due to azetomia.
Serum creatinine- the creatinine is a metabolic product of muscle creatine phosphate- it is a
more sensitive indicator for renal damage than BUN indicators.
Normal values are 50-120mmol/l or 0.6-1.2mg/dl
Serum creatinine will double by 50% with decrease in GFR.
A decrease in GFR leads to an increase in CrCl so renal failure is indicated
Serum creatinine decreases in the elderly
Crcl is the rate at which creatinine is removed from the blood by the kidney. Normal is 80-120
and if
and if its less than 50- it indicates renal disease

Liver function test- LDH- ALP- AST- ALT
LDH- lactate dehydrogenase-
LDH1 and LDH2- heart
LDH 3 lungs
LDH4 and LDH5 liver and skeletal muscle

ALP- alkaline phosphate

AST- Aspartate aminotransferase

ALT Alanine aminotransferase

LDH- this is a glycocyitc enzyme catalyeses interconversion of lactate and pyruvate in most
tissues. LDH present in high concentration in the liver, heart, kidney, lung and skeletal muscles
- LDH is quickly liberated by the liver when diagnosed by trauma, infection or ischemia. So it is
useful for MI, hepatic disease and Lung disease

ALP- Alkaline phosphate- produced from the liver and the bones sensitive to partial to
moderate bile obstruction

AST- Aspartate aminotransferase- also known as SGPT- found in the heart, liver tissues, skeletal
muscles and renal tissues.
ALT sensitive to cell damage and less sensitive than AST.


Cardiac Enzymes
Cardiac Troponins (Tn)
Troponin C T and I are compex proteins that mediate the calcium mediated interactions of actin
and myosin
Troponin T is in cardiac and skeletal muscles
Troponin I is only in cardiac muscle
Troponin C is present in two isoforms in cardiac and skeletal muscle
Troponin T and I are more careful indicators in myocardial injury
They are used to assist in the diagnosis of acute MI


Troponin is a primary diagnostic test for MI

Serum bilirubin (bile)
Breakdown product of rbc and is formed in the reticulocytes and then transported into the blood
Bile is bound to serum albumin
When bilirubin arrives in the liver at the sinusoidal surface of the liver cells the free fraction is rapidly
taken up by the liver and converted to bilirubin diglucorinde and monoglucorinide (conjugated
bilirubin)- This is then released into the intestine.
Most is destroyed in the freaces but some is absorbed in the blood and the liver.
Increase of bilirubin indicates jaundice.
Causes of increased bilirubin :
-increase hemolysis urine colour not changed
-biliary obstruction- bile in urine and kidney stones and dark urine, chlorpromazine gives intra hepatic
cholestatsis
-liver cell necrosis- viral hepatitis- dark urine colour and bile in the urine
Serum proteins (blood proteins)
These are albumins and globulins (alpha beta and gamma)
Albumin is the major glycoprotein in the blood- it binds to acidic drugs
it is produced by the liver
levels are decreased in liver disease e
Albumin contributes of 80% colloidal osmotic pressure and hence hypoalbumineia is associated with
edema and ECF.
Hypoalbuminea
decrease in essential amino acids due to malnutrition can lead to hypoalbuminea
It can be lost directy from the blood because of haemorrge and burns
Hyperalbuminemia
Increase in this can cause shock or volume depletion

Condition albumin Alpha glycoprotein
Renal failure Decrease Increase
Hepatic failure decrease Increase
arthiritis decrease Increase
burns decrease -
pregnancy decrease
Stress/trauma decrease Increase

Globulins ( alpha, beta or gamma)
They are the same as Ig
opposite effect to albumin
Bind to basic drugs

Urinalysis
provides basic information regarding renal infection and function and UTI


Normal urine- clear, pale yellow or golden yellow
Red- usually indicates blood in the urine- or can be drugs such as -phenolphtalein- laxative
Brownish yellow- bile- jaundice
Orange red- rifampicin
Dark urine- metrondiazole and metformine
Blue urine- triamterine
Pyuria and bacteruia are UTI and are normally symptomless.

Drugs that discolour urine
Rifampicin- red/orange and alos body fluids
Metronidazole (dark)
Tetracycline
Riboflavin
Pyrinzinamide
Trimaterin
Nitrofuration
Phenolphatin
Pyrivium pamoate

Specific gravity increase- indicated excessive blood sugar or protein in the urine
decreased SG indicates diabetes insipidus
Fixed SG- kidney loses ability to dilute or concentrate the urine
Protein in the urine proteinurea- can be caused by UTI, renal infection, venous congestion

Albuminurea- indicates glomerular permeability
microalbimurea- - albumin the the urine that is higher than normal but lower than the set
standards- sign of renal nephritis
Glycosuria- indicates diabetes
Ketones
They do NOT normally appear in the urine- and if there are no glucose stores then the
fat stores start to metabolise to form ketones.
Ketonuria is uncontrolled DM or starvation or zero or low carbohydrate diets
Three types of ketone bodies:
-Betahydroxy butyric acid (80%)
-Acetoacetic acid (2o%)
-acetone (trace amounts)

URINE ANALYSIS

Urine Ph- 4.5-9
Specific gravity- 1.010-1.025
Protein levels -50-80mg
Glucose levels 180
Ketone do not appear in urine


Common serum enzymes
CK or CPK is found in the heart and skeletal muscle
CK transfers the high energy phosphate group in tissues that use a lot of energy
Total CK- increases with excersise and IM injections of drugs irritating to tissues like
diazepam

Ck isoenzyme
-Deep IM jnection can increase CK levels
can use to diagnose MI or skeletal muscle damage

-CK-MM in skeletal muscle
CK-BB- brain tissue
CK MB in the heart for MI

Blood work
CBC
Haemoglobin
Hct or PCV- this is the % of the RBC per volume of blood
WBC

RBC
MCHC
Platelet count, reticulocyte count and LDC do not usually need to be included in a CBC

Hct- this sit he percentage of the RBC per the volume of blood. Decrease in HCt is results from
anaemia or bleeding or bone marrow depression. Chronic anaemia or hemolysis
Increase can result from polycythemia

WBC- 4000-11000

Neutrophils 55-75%
Lympocytes 20-40%
monocytes 0-7%
eosionphos
basophils

-Bacterial infections indicate increase in neutrophil and drcrease in lymphocytes
-Viral infection increase lymphocytes
Allergic reaction- increase basophils (asthma)
COPD- neutrophils increase
AIDS- T cell decrease (lymphocytes decrease)
TB- increase monocytes


Warfarin and anticoagulants

Warfarin Heparin LMWH
Oral Iv or SC Sc
INR and PT apTT Not monitored due to
predictable reactions but
monitor rash and bleeding and
heparin assay


Normal INR 2-3
Heparin- apTT, heparin assay and PT

PT- prothrombin is syntheisied in the liver and then is converted into thrombin during the blood
clotting process.
-Thrombin is critical in the homeostatic process because it creates the fibrin monomers that
form the mesh over the wound and activates the platelets
-Clotting time is measured
-PT- measures 2,7,9,10 factors
-Normal value is 10-13 seconds

Increase in PT (INR)
can occur due to inadequate vitamin K in the diet or drugs that incrase PT- warfarin, heparin,
LMWH, high dose of salicycates, and antibiotics.
Higher the INR- more bleeding- thinner blood

Decrase in PT (INR)
due to increase in vitamin K- vegetables and supplements and can cause an increase in blood
clots

IF not bleeding and increase in INR- stop warfarin

aPTT
this measures the intrinsic clotting system which depends on the facors 2 7 9 10
measures the other clotting factors- 8,9,10,12,13
monitored for heparin therapy
normal values are 21-45 seconds

Increase in aPTT
-liver dysfunction
-inadequate vitamin K intake
-poor or inadequate nutrition
-increase in aPTT increases the risk of bleeding


INR>3- overdose of blood thinners warfarin, NSAIDS
INR<2 oral contraceptives , vitamin K

Normal lipoprotein level
-Low density- (LDL) <2.2
-TCG<3.6
-HDL >0.9
-C.HDL 5mmol/L

Thyroid function tests (serum)

Normal hypothyroidism hyperthryroidsm
Serum TSH 0.5-5 >5 <0.5
Serum TSH elevated Serum TSH, Free T4 and T3

Thyroid disease test free T4 and serum TSH

Free T3 and T4
Replacement therapy for hypothyroidism is serum TSH, Free thyroxin index, resin TII uptake
(Rt3U) and TT4

Free T3 and T4

FT4 is the mist reliable test for hyopthyroidsm and hyperthyroidism in contrast to:
FT3 is expensive
Total tt3 and tt4
TT3 and TT4 measure free and bound total serum t3 and t4
TT3 is useful in Graves disease
TT3- not good for hypothyroidsm

Dehydration
BUN normal in mild
Moderate increase BUN
Sever increase BUN, increase Hb and low sucrose










TIPS
Normal K level range is 3.5-5mmol/L
Precursor of plasma cells B lymphocytes
Leukopenia- reduced lycocytes
Normal CrCL- 80-120ml/min
Bilirubin is icnrased in jaundice and AST and ALT AST>ALT and AST more sensitive
Alcholic hepitisi- AST >ALT (both elevated)
INR is normal test for warfarin
INR in warfarin patient should be between 2-3
LMWH- not monitored- monitor for rash bleeding and heparin assay
INR >5 bleeding talk to dr stop warfarin
aptT and PTt is heparin
in MB- CK MB and LDH and Ti
Troponin I is elevated after an MI
Calcitionin opposes action of parathyroid hormone
Hypothyroidism measured by TSH
Hyperthyroidism TT3 and TT4 (free)


14- Nutrition
Canada Health Food Guide
-Carbohydrate 55%
-Proteins 30%
Fats <5%
Fibre 30g/day
Minerals and vitamins
Water 8-10 glasses a day
Salt <2g /day

Allergens
-milk- lactose
Peanuts
Tartazine- colouring agent

Vitamins
Water Soluble- B and C
Fat Soluble ADEK

Vitamin short notes
Vitamin B1- Thiamine
Vitamin B2- Riboflavin
Vitamin B3 niacin
Vitamin B5- panththenic acid
Vitamin B12- Cyconobalamin
Vitamin C Ascorbic Acid
Vitamin E- alpha tocopherol
Vitamin K1- phylloquinone
Vitamin K2 menaquninone
Vitamin k3 synthetic menaquione

Vitamin B1- Thiamine

Contains a sulphur atom
Made from pryrimidine and a thiazole and couple by the methylene bridge
Rapidly converted to Thiamine pyrophosphate ad thiamine diphophosporansferase
Usually in the diet but if not need to take thiamine
Deficiency:
Reduced capacity for the cells to release energy
Can cause beri beri
Wernicke korskaoff syndrome
Found in chronic alcoholics so vit b1 supplement is needed





Vitamin B2- Riboflavin

-Precursor for co enzyme FMN and FAD
-It decomposes when exposed to light
-Can cause deficiencies in new born
-Found in eggs, meat, cereal and milk
-1.2-1.7 day
Seen in alcoholics due to poor diet
-Glossitis, stmatisis, photobhobia and sehorrhea is deficiency

NiacIn Vitamin B3
Nicotinic acid and nicotinamide can serve the source for this
Niacin is required for the synthesis of active form of vitamin B3
NAD and NADP are co factors for dehydrogenases
Niacin can be derived from tryptophan and is not a true vitamin
Nicotinic acid can be used in the diet but is not recommended in patients with gout or diabetes
Glossitis, depression dementia, are the deficiency
Can cause PELLAGRA- depression, diarrhea,
Hartnup disease causes it too
decrease in niacin can be caused by isonazid therapy for TB







Vitamin B5- Pathothenic Acid

- It is formed from Beta alanine and panthtenci acid
- can be gained from wholemeal grains, legumes and meat\
- -Extremely rare

Vitamin B6- Pyridoxine- pyridoXAL

Used as a supplement for nausea and vomiting in pregnant women
0.6mg/day
Drugs can reduce it like anti tb isonazaid, penicillamine for RA , avoid with levodopa- as vitamin b6
causes the conversion of levodopa to dopamine and can give nausea and vomiting
Biotin
this is co factor that is required by enzymes involved in carboxylation reactions- eg acetyl coA
carboxylase and pyruvate carboxylase
-Found in foods and synthesized in the bacteria
-Can be got from excessive consumption of raw eggs-
-Antiboitics can cause decrease in biotin in the long run








Cyanocobalamin- Vitamin B12
Has a corrin ring and a tetrapyrolle ring structre
has a cobalt ion in the center

It is absorbed in the ileum
it is a co factor used in catabolism of fatty acids- and amino acids valine isoleucine and transcobalamin
Megoblastic or pernicious anaemia can occur from the defficeicny
due to the lack of the intrinsic factor that is produced to help it to be absorbed
Should be given parentally- oral supplements are not effective
Oral contraceptives, trifluperazine, KI metformin should not be taken at the same time
Use in the elderly

Folic Acid
COnjucated molecule and has a pteridine structure|
it is linked to PABA
Animals cant synthezise paba so it mst be got from folic or folate in the diet
Yeast and leafy vegetables- are a good source
Megoblastic anaemia and in preg can cause neural tube defect
Conc decreased by- anticonvulsants, oral contraceptives, metformin, dapsone, 5-FU, sulfonamides




Vitamin C Ascorbic Acid


Wound healing- used in collagen synthesis
Fruits and veg
deficiency causes scurvy smokers are vit C deficient

Vitamin A
Has 3 active molecules
Beta carotene retinol retinal retinoic acid
Retinol, retina and retinoic acid
Beta carotene is precursor of vitamin A
Rhodopsin is inactive form of vit A in vision
B-carotene is two molecules than of retinal that are linked at their aldehyde ends also called pro vitamin
A
Good anti oxidants
od vit A is toxic
can cause bone pain- nausea diarrhea and hepatosplegenomy (enlarged liver and spleen)
Def- xeroplhtalmia
Night blindness-
Early symptoms- increased suspectivity to infection, cancer anaemaia
prolonged- deterioration of the eye tissue- and progressive keratinzation of the cornea
Accutaine- cis retinoic acid- oral only
TTT Retin A trans retinoic acid- cream
ov >2500IU
Retinol

Retinoia acid

Vitamin D
Steroid hormone
1,25 dihyoxycolecalcferol is the active form
deroved from ergosterol and prodced in the skin
Chronic renal failure is a deficiency of vitamin D3
Milk- fish eggs liver oil
rickets in children and oesteomalacia in adults
Newborn should get vitamin D drops (oral)

Vitamin E
Alpha tocopherol 0 is the strongest antioxidant
Vitamin $E is a natural antioxidant
Storage site is the adipose tissues
Increased intake is needed in premature infants and persons with fat diet
cause increase in blood cell fragility
Vitamin K (Quinolone structre)
K1(phylloquninone) green vegetables
k2(menaquinone) interstinal bacteria
K3 synthetic mendione- When administed it is alyklated to V2
maintain clotting 2 7 9 10
antidote for warfarin

Essential fatty acids
These include lineloic acid (omega 3), arachdionic acid, eicosopentanoic acid,
EFA- need to be gained from the diet-
Omega 3 and omega 6 is from vegetable oil and fish oil provide eicosapentanoic acid- some EFA can be
made from other EFA like araciondonic acid



TIPS
Vitamin B12- commonly found in animal products
PABA is a precursor of folic acid
Vitamin D deficiency is common in Canada and USA
D3 1,25 dihydroxycolecalciferol is the active form of vitamin D
Supplement of folic acid in pregnancy reduces neural tube defect
Storage form of vitamin D is D2
Skin exposed vitamin D is cholecalciferol
ADEK absorption occurs in the small intestine
All B compex washout except vitamin B12
Folic acid is used for DNA and RNA synthesis
Vit A overdose causes toxicity
Alcoholic patients deficient in vitamin B12
Chronic renal disease need vitamin D
Deficincy in newborn treateted for hyperbilurimea by photo therapy riboflavin
Niacin is not a true vitamin
Pellagra is due to def of vitamin d3
Pteridine ring is in folic acid
Scurvy is in vitamin C def
B carotene is the precursor of Vitamin A
Omega 6- lineloic acid
Omega 3 linelonic acid act like aspirtin antiplatlet
Vit E toxicity prevents vit C from working
Vit B1 is beriberi and Wrnick Korsaff syndrome
Vitamin D in chronic renal disease
Vit b12 has cobalt center- cycobalamin
Vitamin b2 riboflavin- pellagra





Chapter 15
Microbiology

Bacteria: Contain cell membrane and cell organs
Bacterial shape- round(coccus) rod like (bacillus) spiral (spirochete)
Bacterial nucleus is not surrounded by a cell membrane
Bacterial ribosomes are 30s 50s and 70s
Cell membrane consist of cytochrome, lipids and enzymes
Mesosomes are invaginations of bacteria
Plamid- circular piece of dna
Endospore- is a inactive cell
External layer is a capsule and is resistant to phagocytosis
Cell wall- resistant to osmotic pressure
Peptidoglycan cell wall present in gram positive and gram negative cells
Mucopeptide is presnt in peptidoglycan
Techoic acid- water soluble polymer- present in gram positive only
Periplasmic space gram positive bacteria between cell memembrane and outer cell
Outer membrane- grame nevative- phospholipid layer embedded proteins and prions
Lipopolysaccharide- gram ve and consist of lipid A an endotoxin
Glyocalyx is the external layer- slime layer and is adhesive
Appendages- flagella/pilli/ ordinally pillae or sex pillae
Bacteria growth cuvre- lag- increase in size and exponential is increase in numbers
Decline is death phase
Obligate aerobe- generate h202 are a bactericidal
Superoxide dismustase- enzyme released to neturalise h202
Obligate anaerobe- has no superoxide dimusterase
Facultative anerobe- most pathogenic- can switch from fermentative to respiratory metabolism
Aero tolerant anerobes- similar to facultative and remain fementative
Canophilic anaerobe- similar to facultative
Oxygen requirement
Facultative anerobe- most pathogenic can shift from fermanetative to respiratory
Obligate aerobe has superoxide dismustase enzyme and releases h202
Obligage aerobe killed by 02
Aerotoelarnt anaerobe femain fermentative



Virus
No cell structures
Made of DNA RNA and protein
All are harmful

Fungi
cell membrane contain ergosterol layer
made up of thread like structures called chitin
Protozoa
eg amoeba
unicellular or single cell organisms- based on flagelletes-

Atypical bacteria
Mycoplasma: have no cell wall
Rickettsia- can be transported by ticks and mites
Chlymaida- lack ATP synthesis
Mycobacteria
Cell membrane contain a mycolic layer
acid fast test can detect it




Gram ve has more layers than gram +ve
Gram +ve Gram ve
Stain blue or purple Stain pink or red
Techoic acid LPs in the outer membrane
Peptidoglycan layer is thick Peptidoglycan layer is thin
Exotoxin is a metabolic product Endotoxin is metabolic product
Exotoxin is thermoliable- and destroyed at high
temperature. It is a high molecular weight protein
Endotoxin is complex and made of phospholipid-
polysaccharide and protein



Gram positive Gram negative
Cocci- Streptococcus
S pyogenes (A)
S agalactiae (B)
S bovis (D)
S pneumonia
S viridians

Gram ve cocci
NISSERIA
N gonorrhea
N meningitis
Moraxella catarrhalis
Stapylococcus
S aureus
S epidermis
S saprophyticus
Enterococcus

Rods
E coli
Klebesiella pneumonia
Enterobacter
Baccili
C. dipthariaie
Listeria monocytogenes
Norcardia
Baccilus cerus
Shigella
Proteus mirabilis
Salmonella

Gram +ve and anaerobic
S typi
S eneridis
Vibero cholarea
Clostridium P aeruginosa
Hinflenza
Y pestis

C perfinges, Gram ve anaerobic
Fusobacterium
B fragilis
Bacetiodies
C difficile, C botilium, C tetana



Gram +ve cocci
S aureus
Catalase and coagulase
positive
Found on the skin and in the
nose
On boils
Wound infections
Toxic shock syndrome
Penicillin g and penicillin v
Beta hemolytic streptocci s
pyogenes
Tonsillitis, cellulits,
septecaemia can occur in
immue diease like rheumatic
fever
Clarithromycin
Azithromycin
Erythromycin
Penicillin G
S pneuomanae CAP, ototis media and
menigitis
Amoxicillin otitis media
Penicillin G
Clarithromycin
Azithromycin

S viridians Endocardiits and dental
carries
Penicillin G or amoxicillin
S epidermis Instrument contamination-
cathether infections UTI

Gram ve
Cornyebacterium diptherai


Diphtheria disased due to
toxic production
Erythromycin or penicillins
Tetracycline

Cl tetani
Cl perfugines
CL botilium
Cl dofficile
Tetanus
Gas gangrene
Botulinism
P colitis
Vancomycin or metronidazole
Gram ve cocci
Nessesieria menengitits

Nesseria Gonorrhea
Menigococcol meningitis- and
shock of URT
STI- always pathogenic
Penicillin G

Cephalosporins 2
nd
or 3
rd

generation or ciprofloxacin
Gram ve bacilli
E coli
Proteus
klebsiella
UTI
Travelers diahreaha
Wound infection or sepsis
Inhabitants of the gut
E coli (sulfa drugs) UTI
E coli diahrrhea ciprofloxacin
S typhi
salmonella
Enteric fever, food poisoning,
most Sp are animal pathogens
and typi only infects man
Chloramphenicol and
ciprofloxacin
Shigella Dysentry Ciprofloxacin
P aureginosa Hospital acquired and
opportunittic infections
Aminoglycosides
3
rd
gen cephalosporins-
crefazidime or cefurtoxime
Ampicillin
H influenzae Pneumonia, meningitis, otitis
media
Clarithromycin
Azithromycin



Acid fast bacill
M tuberculosis Tuberculosis- most common
infectious cause of death
world wide
Isonazaid
Rifampicin
Ethambutol
Pyrazinamide
Streptomycin
M leprae leprosy Dapsone
Rifampicin
Spiroches
Treponema pallidum Syphilis (STI) Doxycycline
Penicillin G
Borellie burgoferi Lyme disease- tick bourne
infection causing rash
arthlagian and nerulogical
signs- bull eye rash
Tetracycline



Fungi Have thick ergosterol
containing walls- and grow in
humans as budding yeast cells-
and slender tubules hyphae

C. albicans

Thrush , valvovaginitis,
mucicutaneous infection
Nystatin
Clotrimoxazole
Miconazole
T Pedis Atheltes foot Clotrimoxazole
Tolfinate
Dermatophtes Ringworm Acquired from animals
sometime
Skin naiks hair
Aspergillus Allergic reactions or
opportunitsitc infections and is
airbourne



C neoformans Meningitis in
immunocompromised= soil nd
pigeon droppings



Amoxicillin
Tetracycline


Protozoa
Plasmodia Malaria- Chloroquine, mefloquine,
primaquine, doxycylcine

g. lambalia Low grade GI disease-
giardiasis
Metronidazole
E. Histolytica Dysentry- severe when it
spreads to the liver
Metronidazole, Ciprofloxacin,
Cotrimoxazole
Viruses
DNA virus
Adenovirus Conjunctivitis- sore throat-
Herpes simplex
Herpes Zoster
Cytomegalovirus
Epstein BAR (EB virus)





HSV1 and HSV2 can cause oral
and genital lesions.
HSV 1 is cold sores and
kertaoconjunctivitis
HSV 2 is genital

VZV can cause chicken pox-
shingles glandular fever
rosella infantum (6
th
disease)
Aciclovir
Famciclovir
Foscarnet
Ganciclovir

Hepandavirus Hepatits B Transmitted via sexual or
bodily fluids
Interferon Alpha
Papilliovirus

Polyomaviurs
Warts and cervical cancer

Hamaerogic cysts
Garadsil used for cervical
cancer

Poxovirus Smallpox
Orthyomyxoviruses- Influenza
A and B
Influenza (flu) Amatadine
Neurominidase inhibitios
Osetlamavir
Zanamavir
Rimantidine (influenza A )
Flavivirus- yellow fever-
hepititis
Yellow fever chronic hepititis
Paramyxoviruses- Enterovirus-
RSv measles mumps
Resp infections Croup Can be severe in infacnts
Reterovirus Rhinorrhea- hepatitis
common cold
Rhino- runny nose
Rotavirus gastroenteritis
Reterovirus HIV HTLV Aids T cell leukaimea NRTI NNRTI protease
inhibitors

Rhabdovirus- rabies rabies Zoonotic infection








Togavirus Rubella
Alpha virus
German measles
Encephilitis


Bacterial infections
Stye- external hordeulu or internal hordelum

Causes- S aurus

Site of infection- edge of the eyelid- and head
ruptures within a few days

Treatment
Warm compress and cloxacillin or flucloxacillin

Inclusion and drainage are indicated and
patient should be referred if they do not
respond to the treatment


Conjuncitivits Pink eye or red eye
Viral- bacterial- chlyamidial
Non infectious- foreign or allergy

Treamtnet
Oral tetracycline doxycline
Erythromycin safe in pregnancy
Azithromycn 1g as a single dose
Amoxiciilin safe in pregnancy
Ceftraixone- 1g IM single dose Genococcal
conjuncitivits in adults

Purleunt or mucopurleunt discharge is
bacterial
Viral non mucouperulent discharge

Watery discharge can be due ti URTI or
adenovirus

Viral conjunctivitis is follicular reaction and
prerucicula lympadenothpy

Viral conjunctivitis
Treatment is supportive- topical corticosteroid
therapy is controversial

Children kept out of school for 2 weeks
following onset of infection

Bacterial conjuncitivits
Staphylococcus and strepoloccus
H influenza in children
Gentamicin or tobramcyin eye drops
Fusicidic acid eye drops

New born conjunctivitis
c.trachomitis or n gonnorhea
chylamidia trachomonitis- erythromcycin
n gonnherea- ceftriaxone
2.5% povidone iodine is best for the
propylaxis




Cannilcuitis
Actimces
Mechanical expression of exudative or
granular material from the canculi and
penicillin G
Refer the patient for a definitive treatment


Darocystitis
Streptococci or s pneumonaie
Infection of nasoscrlimal sac
Acute- amoxicillin
Chronic irrigation nd penicillin G


Keratitis
Bacteria fungi herpes simplex acanthameobea
Sight threatening and so an emergency

Herpes simplex keratitis
HSV 1
Topical disease- acyclovir 5 times a day and
continued 3 days after healing
Trifluridine and hifluridine drops
Stromal disease- antiviral and corticosteroids



Ear nose and thorat

Common cold Rhinovirus

Sinusitis Bacteria s pneumonia
Pharngitis sore throat Virus adenovirus
Bacteir s pyognees
Pneumonia H pneuomina
Bronchitis H influenza
Otitis media S pnenonia h influenza m catrrahlis

Skin and soft tissues
Cellulitis
S pyrogens or s aureus
Dermis
Penicillin and flucolaxcillin
Acute spreading of the dermis- lesion is hot
and red and swollen
Impetigo
S aureus and S pyogenes
Bullous crusted pustular eruption of the skin
Due to invasion of local causative agent
Penicillin or amox and fluclox

Folliculitis (boil)
S aureus
Penicillin or amoxicillin

Erysipelas- rapidly spreagind infection fo the
skin
S pyrgiens or s aureus
Dermis of the face
Penicillin or amoxic with or without fluclox

Neroctising fasicits
Inflammatory response of tissue below the
dermis and spread along the facial planes and
disrupt the blood supply and cause necrosis
and gangrene
S pyogenes (B haemolytic group)
Benzyl penicillin and clinamycin with or
without metrondizaole

Gangrene
Coliform or sterptococco (group A)
Penicillin and celhaliposin beta lactam and
qunilones can be used
Pai fever systemic toxicity

Viruses
Papillonoma virus
Molluscum contagiosum (pox virus)
Pox virus from sheeps or goats



Common wart
Fleshy papule
Papovascular lesions with systemic spread like
herpes simplex

Fungi
Dermatophyte Ring worm or skine lesions (keratin loving
fungi) C neoformans, or Blastomyces
dermtitidis
Tinea pedis Atheltlets foot clotrimazole miconazole
tolnafate

CNS infections
Neonatal (6 weeks) group B strep E coli and other
Ampicillin + gentamicin or ampicillin and
cefrtiaxone
Children > 3 months and adults
S pneumonia N meningitis H influenza type B
Cefrtoixone or ampicillin and vancomycin

Eldery alcoholics immunocomprimesd E cloli S
pneumonia
Cefrtixione or ampicillin or vancomycin
Meningococcal infection N menignigitis Respiratory route

Haemophilus influenza type B Affects 6 month to 5 year old children
Hib vaccine spread
Blood-> meninges

encepelitis Viral infection of the brain cns and cells
HSV 1 is most common CMV rabies mumps

polio Enterovirus
TB meningitis Meningococcus Pneumoccus H influenza
CNS
Benzyp penicillin menicoccois
Bezyl penicillin/cefotaxime or vancomycin if
resistant for pneumoccus
H influenza cefotaxime
Prop meng rifampicin or ciprofloxacin whole
family or close contacts

Respiratory infections
CAP Causative agent: S pneumonia (most common)
Abulatory patients 18-40 year
-M pneumonia (24%)
-S pneumonia (5%)
-Chlyamidia Pneumonia (2%)
-H influenza (1%)
-Legionella pneumophilia (1%)

Requiring hospital administration
S pneumonia (17%)
M pneumonia(14%)
Chylamidia pneumonia (10%)
H influenza (7%)
L pneumophilia (1%)
Emergency treatment is penicillin V
S pneumonia amoxicillin, penicillin G or
macrolide
H infleuzna 2
nd
or 3
rd
gen cephalosporin +
clauvnate
S aureus-
Methicillin susceptible- Cloxacillin
Methicillin resistant Vancomycin
M pneumonia and C pneumonia- Doxycylcine
or macrolide
Legionella- Fluroquinolones- marcolide
rifampicin
E coli- aerobic gram ve bacilli- 2
nd
or 3
rd
gen
cephalosporins- initial therapy should be with
cefoxitin pipercillin or tazobactum

Brochitis Based on age group
<1 year- RSV , parainfuenza , corona virus
1-10 years- parainfluzna , enterovirus rsv
>10 years- influenza RSV adenovirus

Bacterial bronchitis Chlyamidia pneumonia , mycoplasma
pneumonaie `
Treatment Routine antibitict not recommended
Atipyrectci or angalestic aciteomenophen
Antiussive- dextromethorphan
Beta agonist salbutamol
`

Urinary Tract infection
Lower UTI Cystitis and urethritis
Upper UTI Polynephritis and uretitis

UTI most common agent is E coli
Cystitis Cotrimoxazole, or amoxicillin or ciprofloxacin
Urethritis Urea plasma and chlymidia infection-
doxycylcine during pregnancy use
erythromycin
Pyleonephritis Bacterial infection of kidney subustances
Amoxicillin cephalosporins cortimoxazole or
aminoglycosides


Sexually transmitted infections
Causative agents
Nisseria gonorrhea
Chylamidia
Lymphogranuloma
Bacterial vaginosis
AIDS
Hepititis B and C
Chancroid (syphilis)
Genital herpes
Genital Warts
Candidia or yeast are not STI

Candidia is not STI so no treatment and if the
symptoms recur within two months then the
partner would need treatment





Infectious Arthiritis
Lyme diease
Oseteomyleitis are all bone and joint
infections


Infectious arthiritis

The gonococcus bacteria can cause different
symptoms in males and females

Women develop red sores on the hands and
feet and severe pain in the wrist and ankles

In men gonococcus only affects a single joint
and most often the knee is affected.


Arthiritis is due to gonococcus and oral
ampicillin is the treatment

Surgery doesnt really help

Lyme disease
Is tick bourne and can cause arithirits and in
severe cases heart and cns complications

Spirochete (Boreli bergodorferi) Is transmitted
to humans via deer tick and prevelant during
summer months
Tetracycline is the drug of choice
Oesteomylitis Bone infection of the marrow and caused by S
aureus

G I infections
Stomach H pylori is the common infection and is
associated with peptic ulcer disease and
gastric cancer
Large intesting 99% anaerobes
Bactersisides- Bifidobacterium clostridium
anerobic cocci and anaerobes
Anerobes enterobacaea- e coli enterococci
proteus
Food poisoning
Shigella Dysentry travelers diaorrhea
C jejuni Travelers diahrea
Salmonella Eggs meat poultry and travelers diarrhea
C difficile P colitis
E coli Meat poisingig and travelers diarrhea
S aureus Meat mayonnaise custard
c. perfigines Actue gastroenteritis and reheated dishes
Norwalk virus Diarrhea in hospitalized patients
Entomebea Ameobiasis
B cerus Reheated rice
V parahemolyticus Contaminated sea food
Listeria Meat


TIPS
Beta haemolytic bacteria- Sterp. Pyogenes ( Group A strep_
Toxic shock syndrome- S aureus
Lyme disease is borrelia burgodorferi
Techoic acid- is in gram positive bacteria S pneumonaia, S aureus
Encephelitis is a brain infection caused by:
-Viral alpha viruses- the ones that cause rubella or rabies virus HSV virus
-bacterial- N meningitis
Chylamidia in new more is C trachomitis-
Non gonococcal infections that cause UTI is E coli and chylamidia
Diphtheria- Corynebacterium diphteriae, URTI
Sypiliis- Treponema palladium
Anthrax- bacillus anthraics
E coli- gram negative anaerobic cocci
The bacteria that degrades h202 is obligate anaerobe S aureus- s epidermis does not
Live vaccines- MMR, sabine, small pox, VZ, yellow fever
Killed vaccine- polio, influzna rabies
Viral diarrhea- rota virus
CAP- S pneumonia ambulatory patients M pneumonia. And Hospital admission is S
pneumonia
Sub acute endocaritis- s aureas and s viridians
Syphillis Tropenem Pallidum
Gram +ve bacilli=
B diarrhea- C jejuni, E colli shigella
Otitis externa- pseudomonas
Aspiration of ear is in middle ear infection
Encephalitis HSV
Gram +ve bacilli C diptheri
Sinusitis H pneumonia, H inflenze M carttharalis
Nocosomial infections S aureus and Pseudomonas
Shingles herpes zoster
Herpes- HSV 1 HSV 2 VZV CMV Epistein barr
Tuberculosis:
caused by Menigicococcus, Pneumoccocus, H influenza
Menigcoccus- benzyl penicillin
Pneumococcus- benzyl penicillin/ cefotaxime/ vancomycin if resistant
H influanzae- Cefotaxime (start with penicillin + cefotaxime)
Propylaxsis- Rifampicin or ciprofloxacin

Pneumonia
emergency treatment- Penicillin V
S pneumonia- Penicillin G- amox or macrolide
High level resistant- quinolines and 3
rd
gen cephalosporins
H influenza- 2
nd
3
rd
generation cephalosporins or amoxicillin and clavanate
S aureus- Methicillin Susceptible- Cloxacillin and meth resistant vancomycin
M pneunonae or C pneumonia- Doxycycline or macrolide
Legionella florquniolines macrolide and rifampicin
Ecoli- 2
nd
nd 3
rd
cephalosporins- initial therapy- cefoxtin or piperacollin and
tazobactum

16 - Cell and molecular Biology
DNA replication
This first occurs by separating the double stranded DNA molecules and then each new strand then
serves as a template onto which new bases are paired upon in complementary base pairing. The same
thing is done in transcription from DNA t RNA. The RNA is the intermediate between the DNA and the
protein.
Cell organelles
ER membranes that extend throughout the cytoplasm . only in eukaryotic cells. Cytochrome P450 is
present in ER
RER- helps in protein synthesis
SER- helps in lipid synthesis
Golgi bodies contain sacs help in protein and lipid synthesis
lysosomes help in digestion
Genome- complete set of DNA in the organism
Gene expression- gene is DNA sequence that encodes a protein or RNA molecules

Nucleotide- Base+sugar+phosphate
pentose sugar- ribose or deoxyribise
phosphate 5 carbon
nirotgenous base

Nitrogen bases
Purine bases adenine and guanine
Pyrimidine bases- thiamine- uracil- cytosine

at (2 base pair)
cg (3 )
Au
Purine 2 bonds and pyrimidine 1 bond
cDNA- complementary DNA that is syntheisezed from RNA rather than from DNA template
used in cloning or to locate specific probes\
RNA differences has Uracil instead of Thiamine, single stranded, has ribose instead of deoxyribose
RNA polymerases- enzymes help in the synthesis of rRNA tRNA and mRNA
3 types of RNA based on their functions:
rRNA- ribosomal = 80% is syntheisie in ribosome in cells
tRNA- 15% of tota- tRNA amino acid carries the specific amino acid to the site of protein synthesis and it
contains an anticodon that recongnizes al codons on mRNA
mRNA- 5% carries the genetic information from DNA to cytosol in protein synthesis
Codon
present in MRNA nd is have a set of 3 bases- there are 64 combinations but 20 common amino acids
UAG UAA UGA- dont code for anthing
When UA UGA UAG is reahed its nonsense codon and it stops
Step 1-> transcription
Step 2 -> translation
DNa mrnatrna rrna protein
Transcription 1
st
process of protein synthesis and DNA is copied into mRNA
from mrna to dna s reverse transcriptase
Translation- mrna is brought to the ribosme by the trna and then under goes complimentary base
pairing catalysed by amino acyl trna synthase- and then it keeps going until a stop codon is reached.
After which there is post translational modification glycoslyation sulfation etc splicing of exons
Intron and exons
Coding reigon of a eukaryotcyte gene is introns and exons- intron is a segment of a gene that is suited
between exons.
It is not responsible for the coding of a protein- so they wud be spliced out of the mRNA- the exon is a
nucleotide seuqnece in DNA that carries the code for the final mRNA molecule- and so defines the
amino acid sequence
Remoiving introns is called splicing-

DNA recombinant technology
Cloning
Duplicating DNA and it is identical using a vector
Insert the DNA into a plasmid vector- it is usually an antibiotic resistant getna and recombinant molecule
is formed. Plasmids and DNA fragments must have compatible RE ends for ligation by DNA ligase- The
ends are joined together to form the circular DNA product .
Once the Recombinnt (circular DNA ) is formed- it is then inserted into a host cell like E coli and allowed
to replicate and then the memebrane of the cells are made permeable with shock treatment of calcium
ions to allow the cells to take up the DNA pieces
So the cells are grown in an antibiotic medium and only the resistant cells will grow
Select the cells with the DNA x and the antibiotic resistant cell use beta galactosidase to get cells with
the DNA x and not the vector due to unsuccessful ligation.
Kill the cells and extract the DNA X
Eg Cystic fibrosis etc

DNA recombinant technology
Plasmid circular piece of DNA used in recominbation and cloning
Endonuclease enzymes- Restriction enzymes are a set of enzymes that are bacterial and they cut the
DNA at specific sites called the restriction sites- by breaking the phospohidester bonds- between the
nucleotides- to form the single stranded DNA they are isolated from bacterial species- and break
foreign DNA
Lyase and lygase are the enzymes
Lyase- splits DNA on specific sites
Lygase- joins the DNA on specific sites

TIPS
Nucleic acid- DNA and RNA
Building blocks are nucleotides
Purine bases are adenine and guanine
Pyrimidine bases are thiamine cytocine and uracil
Base only in RNA uracil
All genetic information in an organism is called as genome
DNA sequence that encodes a protein or a DNA is a gene
Fist step in protein synthesis is transcription
Segment of gene between exons is intron
Split DNA on specific sigte DNA lyase
Join DNA DNA ligase
Small circular chromosome of DNA is plasmid
Genetic disease with chromosome X is haemophilia
Geneome all the genes etc
Microtubule in cell division- centrosomes
Prokaryotes- have cell membrane
Eukaroycte- have cell wall
Nucleotide is sugar phosbate and base
DNA- A G C T
RNA A G C U
Complementary- AT CG AU
Transcription is DNa to mRNA
Translation is mRNA to tRNa
Andicodon on tRNA
cDNA is complementart DNA syntheisized fromm RNA rather than DNA
rRNA- 80%
virus have single stranded DNA
RNA polymerase is in transcription mRNA
DNA transferase cataylses moving of specific functional grouos such as phosphate
DNA isomerase- isomeraisation
DNA lyase- cutting apart
DNA ligase joining together
Reverse transcriptase is used to make DNa from rna it is a RNA dependent DNA
polymerase
NNRTI- reverse transcriptase inhibitiors and it is a virus
DNA synthesis by reverse ranscriptase- inhibits by AZT
Hapten is a low MW compound that act as an immunogen by chemically complexing to
a large molecue at the cell surface
PCR is used to make a bigger DNA chain or to amplify it- can be used in finger printing
1
st
step is denaturation and the DNA heated o 96 0C to break it up done for 9 minutes
and oly used in DNa that need heat activation
Denaturation is then used heating to 96-98 degrees for 20 -30 mins so DNA double
strand is broken and as hydrogen bonds are broken
Annealing step- lower the temp to 50-60 degrees to allow the bonds to reform between
the base pairs
Elongation depends on the tem and Taq polymerase is used-
Extension/elongation step: The temperature at this step depends on the DNA polymerase
used; Taq polymerase has its optimum activity temperature at 7580 C,
[12][13]
and
commonly a temperature of 72 C is used with this enzyme. At this step the DNA
polymerase synthesizes a new DNA strand complementary to the DNA template strand
by adding dNTPs that are complementary to the template in 5' to 3' direction, condensing
the 5'-phosphate group of the dNTPs with the 3'-hydroxyl group at the end of the nascent
(extending) DNA strand. The extension time depends both on the DNA polymerase used
and on the length of the DNA fragment to be amplified. As a rule-of-thumb, at its optimum
temperature, the DNA polymerase will polymerize a thousand bases per minute. Under
optimum conditions, i.e., if there are no limitations due to limiting substrates or reagents,
at each extension step, the amount of DNA target is doubled, leading to exponential
(geometric) amplification of the specific DNA fragment.
Final elongation- is the single step and performed at the end to ensure the last DNA is
fully extended done at 70-74oC for 15 mins
Final hold: This step at 415 C for an indefinite time may be employed for short-term
storage of the reaction
_______________________________________________________________________

Chapter 17- Pharmacogenetics
This is when genetic differences in individuals can affect the way that the drugs work or are metabolized
in their bodies and the drug response
This allows drugs to be made that are customized to each persons genetic mark up and they used for
cytochrome enzymes that are responsible for drug interactions.
The first step is a detailed analysis of teach of the genes un a single nucleotide polymorphism
SNP- single nucleotide polymorphism- is used when one base pair on a nucleotide replaces another- A
single base differences exist between individuals- this is the most commogenetic variation In DNA.
Defective splicing- is which an internal polypeptide segment is abnormally removed and the ends are then
joined.
Gene therapy
This is the use of genes to cure diseases by altering a single defective gene like in cystic fibrosis
Eg drugs used are HER2 receptor genes and traszumatab used for breast cancer-
HER2 Rna inhibitor

Immunology
In organ transplant rejection due to infiltration of T cells
Most common antibody is IgG
Munorab ad Monoclonal antibody attack CD4
igM first produced when exposed to an antigen
igE activated in asthma patients
acute graft rejection is type 2
tuberculin test is type 4
infliximab is for RA and chrons disease
hapten is low molecular weight molecule that covalently binds to a larger molecule
infliximab is iv
hashimoto is hypothyroidism and is type 2- organ specific autoimmune diease
acute graft rejection is type 2
igE mediates type 1
poison ivy is type 4
neutrophil- respond to bacterial and fungal infection
monocytes hightest cell in the body
basophils not mast cells
penicillin hypersensitivity type 1
lupus drugs HIPP MCQ- hydralazine- isonazid- procanamide- penicillamine- methyldopa-
chlorpromazine qunidine
SLE is type 3 non organ specific autoimmune
Steven Johnson- od of pshycotics or TCA- rash- photosensitivity feverer 3
rd
degree burns
SASPAN- sulphonamides phenytoin allopurinol nsaids sulfoxazole
Red man syndrome vancomycin
Gluten in wheat and rye
____________________________________________________________________________


Immunization
High risk for flu jab- 6month- 2 year kids, egg allergy, asthma, cvd, pregnancy and seniors
Who should not take- < 6 month- egg allergy and flu symptoms
Live vaccines not given to pregnant and immunocompromised
Live vaccine eg MMR, varciella, yellow fever, small pox- sabine polio
Flu vaccine is killed vaccine and effective against influenza a and b
Killed vaccine- flu, hepatitis A and B, thypoid, cholera
Travellers should take Hep A and B
Hep C is chronic hepatitis
Hep B vaccine protects against Hep D
Dukoral oral vaccine is for E coli and cholera- - two doses one week apart and the last one a week before
you travel- High risk latin America- south east asia and Africa ( basically poor countries)
Giadardisil is used for papilloma virus
Biotechnology
-Anameia with chemotherapy- chronic renal disease- erythropoietin
-anaemia for cancer chemotherapy and chronic dialyisis and AZT- epoeitn alph and darpoeitn
-Neutropenia associated with chemotherapy- filgristim
-TNF alpha and beta- enterecept
-Graft rejection- muromonab
-Chrons and ra is infliximab
-haemophilia A def of clotting factor 8
-muromonab attacks CD4 cells
Vaccnes
Ascorbic acid can detoxify the urine
Conjcation reaction is the one that causes acetominphen toxicity