Beruflich Dokumente
Kultur Dokumente
IN PHYSIOPATHOLOGY
AS BASIS OF
GUIDED CHEMOTHERAPHY
With Special Application to Cancer
EMANUEL REVICI, M.D.
Scientific Director, Institute of Applied Biology,
New York, N.Y.
Chief of Dept. Of Oncology, Trafalgar Hospital,
New York, N.Y.
D . V A N N O S T R A N D C O M P A N Y, I N C .
TORONTO
LONDON
COPYRIGHT 1961, BY
EMANUEL REVICI. M.D.
Published simultaneously in Canada by
D. VAN NOSTRAND COMPANY (Canada), LTD.
CONTENTS
FOREWORD
THEORY A ND FAC TS
EW OTHER PATHOLOGICAL CONDITIONS have aroused, as cancer has, the interest of so
Meanwhile, normal development of cancer research has been hindered, sidetracked from its logical course. While thousands of scientists with almost unlimited funds
at their disposal are presently using the most advanced methods for the acquisition of
details, almost no attempts are being made to resolve basic problems, although the cancer
investigator is continuously obliged to realize the dearth of fundamental knowledge.
If we attempt to analyze this abnormal situation further, we can nd indications
that it may have its origin also in a distortion of the proper relationship between the two
factors that, together, make for progress in researchideas and experiment.
The experimental approach provides precise information about particular
phenomena under dened conditions. The analytical method tries to investigate reality by
recognizing the proper place of the various constituents of a whole, the parts being
identied as such by the experimental ndings. On the other hand, the conceptual
method not only provides an inkling of what the completed whole will eventually look like,
but also attempts to predict the properties and the relationship of the component parts.
In dealing with a highly rened and complicated subject, the analytical method by
itself appears inadequate. For example, in atomic physics, the results of experiments are
expressed by numbers giving the values of certain physical quantities that have been
measured. In order to complete the analysis, we must simultaneously determine the
numerical values of certain quantities dening the material bodies, the objects of the
experiments. This is prohibitive so far as canonical coordinates by Heisenbergs uncertainty principles are concerned. With experimental knowledge somewhat curtailed, theory
at present must attempt explanation.
In other areas as well, experiments present only limited numerical values pertaining
to some physical quantities. Were we able to measure all quantities, we could analytically
reconstruct the entire theme of the physical reality. However, when some quantities
cannot be simultaneously determined, this direct reconstruction is not possible and
experiments merely give an indirect approach to what we regard as reality.
If the inadequacy of the analytical approach by itself is evident in the highly positive
disciplines, such as in the physical sciences, it is even more so in biology. As Bohr and
others have intimated, the conditions of uncertainty seem to be much more pronounced
in biology than in physical science. The fact that experiments in biology give only
fragmentary and unrelated results is not surprising; the need for a synthetic theoretical
method in this eld is clear.
In medicine, which is applied biology, the need for the conceptual approach is
especially profound. lt is true that this approach, as the sole approach, has shown its
inherent weakness in the past. There was a time in the development of medicine when
available data were so scarce and unreliable, and the need for ideas to provide some sort
of guidance was so great, that the worker resorted to broad imagination, using it to
replace almost entirely any other form of investigation.
Largely as a reaction to the high proportion of speculations prevalent in the early
immediate skin and systemic effects. Now, as earlier, with few exceptions, the benets of
radiation are no more than temporary. Longlasting good effects still are limited to only a
few radio-sensitive tumors. The resort to isotopes, in which the scientic world has put so
much hope and millions of dollars, also has proved greatly disappointing.
Of the thousands of cases of various kinds of cancer in which isotope therapy has
been tried, only a very limited number of cancers of the thyroid have responded. Not only
because of its continuing failures, but because of its inherent qualitative inadequacy,
radiation does not appear, any more than surgery, to represent the solution for the
problem of cancer.
With surgery and radiation therapy incapable of resolving the problem, more and
more research workers have turned their efforts in other directions. The existence of some
cases of spontaneous remission has led many investigators to believe that immunological
procedures related to cancer would be able to resolve these problems. Unfortunately the
existing knowledge in this specic eld is too meager to permit more than some tentative
investigations, usually only repetitions of similar researches made many years ago with
limited success. Fruitful development of this approach would have to follow the normal
pathway, starting with more knowledge of the complex immunological processes
intervening in cancer.
An enormous amount of cancer research in recent years has been directed toward
chemotherapy. It is a fact that many agents and groups of agents have shown the capacity
to inuence tumor evolution. However, each has had limited usefulness. Results of
treatment have been characterized by inconsistency. Even in seemingly susceptible types
of cancers, results have been good in one case, poor in another and have varied even for
the same patient at different times. The inability to explain and remedy these variations
has discouraged many workers. Although it appears evident that the source of
discrepancies resides in the patients themselves, the general tendency among researchers
has been to try to resolve the problemby nding agents able to act independently of any
differences which exist between subjects.
In despair at the lack of progress in this approach, many workers today are using
the screening enterprise mentioned above as a kind of last resort. For this project, they
have renounced the scientic concept that pharmaco-dynamic activity must serve as the
basis on which an agent is to be tried in therapy. They have fastened into a purely empiric
approach. Now, all available chemical substancesand many others which will be
synthesized especially for the purposeare to be screened indiscriminately, for their
effects on animal tumors with no reason for this test other than that the agents are, or can
be made, available. We will not dwell here on the assumption that routine technique is
more likely than imaginative brainpower to resolve the problem of cancer. The results of
this screening to date have shown it to be an invalid procedure, as expected by most
critical workers. With tens of thousands of substances already tested, the busy screeners
are obliged to recognize that the approach itself is fundamentally erroneous. Experience
has proved that an agent can be wonderfully effective against one tumor and still be en-
tirely inactive in others. Of tens ofthousands of agents tested, less than a hundred have
shown effects on tumors in animals. None appears to have signicant value when applied
in humans. These results have emphasized again the importance of factors other than the
agent itself. One factor lies in the differences which exist between various tumors. Some of
the other factors include variations between species, between individuals of the same
species, between origins of tumors, between spontaneous and transplanted tumors, and
even variations in anyone individual at different times.
Faced with this situation, some workers have concluded that not one treatment but
at least hundreds of different treatments must be found in order to cope with the huge
variety of conditions.
Taking cognizance of these considerations, it has seemed to us that a more realistic
and logical approach is to try to understand the nature ofthe existing differences and to
attempt to make the treatment adequate on the basis of that understanding. It has been
this approach which has been followed in our research.
We have studied the problem of cancer for the last thirty years from an entirely
different vantage point than that used by other workers. Attention has been focused on
the physiopathological aspect of cancer, on the basic changes that occur in the different
patients, with the ultimate aim of understanding the part played by these changes in the
response of cancer to therapeutic attempts. This emphasis on the physiopathological
aspect of cancer has been made possible by applying a more general overall idea of the
nature of the disease.
This approach is based under various new concepts. They concern,
1) The role of the organization in the pathogenesis of the conditions.
2) A dualistic systematization of the manifestations related to normal and
abnormal physiology.
3) The predominant intervention of certain constituents such as lipoids and
chemical elements in the induction of the opposite manifestations.
4) The possibility to integrate the occurring processes into a system of defense
mechanism against the noxious inuence exerted by the environment.
Many general and special problems of physiopathology, some of them concerning cancer
and other conditions, have been analyzed in this framework.
The application of this approach to therapy has resulted from a logical
development of that approach. The recognition of the intervention of a variety of
pathogenic factors, not only differing from one subject to the other, but even changing in
the same subject during the evolution of the condition has emphasized the need for
individualized therapy. As opposed to the tendency to overcome the differences existing
between individual subjects through a standard therapy, the guided therapy utilizes the
knowledge of the occurring different pathogenic particularities in order to correct them. A
high degree of exibility in the treatment has appeared necessary.
As part of this approach to therapy, has appeared the need for more complete
knowledge of the existing differences and their interpretation in terms of the pathogen-
esis of the condition. The search for adequate analytical tests has thus represented the
rst task. The development of day-by-day analysis of the condition has been possible by
choosing relatively simple but reliable procedures. The information they offered was used
to determinethe nature of the agents able to correct with a certain specicity, the
encountered pathological conditions. These two parts the recognition of the existing
condition and the adequate agents, have concretized this approach.
These considerations explain also why the new developed guided therapy cannot
be understood and correctly applied without a sufficient knowledge of its
physiopathological and pharmacological basis. These same considerations have led us to
present the research concerning this approachas a block, instead of fragmented
communications. The form of a monograph has appeared consequently the best suited. In
a further effort to achieve a cohesive presentation, we have separated from the text most
of the technical and experimental data, and presented them as notes at the end of the
text.
ACKNOWLEDGEMENT
Progress in our research has been made possible only through the day by day
cooperation of different groups of co-workers who have contributed years of assiduous
work. Many of them are mentioned in the following pages where the research in which
they took part is presented. I am deeply indebted to them.
I wish to thank all those friends whose personal efforts or, who through their
organizations, have given their material and moral support to the continuation of our
research. Special thanks go to Mrs. Sherman Pratt for her tireless efforts on our behalf.
My sincere appreciation to those who have helped me in the preparation of this
book, Mr. L. Galton, Mrs. E. F. Taskier and Mrs. H. Kennedy for the editorial work; Mrs. E. F.
Taskier for the gures; and Mrs. P. Berger, Mrs. B. Doctors and Mrs. M. Prikasky for the
secretarial work.
I wish to gratefully acknowledge Miss Fanny I-Ioltzmanns devoted friendship and
help.
CHAPTER
t s A S I CC
] O N C E P T SO F
ORCANIZATION
nr\
rNrELLEcruALMEcHANTsM
I
usedby man to acquireknowredgehas
"t
led him to recognizethe existenceo[ relationshipsbetween the various
manifestationsencounteredin nature. He has employed abstractionand
integration to build up conceprualindividualitieswhich he identified as
separateentitiesin nature. Structuralcharacteristicsand dynamic properties have appearedto be the most suitablecriteria for defining theseentities. However,curiosityhas constantlyimpelledman to attemptto extend
his knowledgeby explainingand correlatingtheseentities,and an important means has been analysesbreaking them down into their component
parts.
Out of these analyseshas come recognitionof the fundamental importanceof organization.F^ot,as entitieshave been analyzedone after the
other, it has becomeclear that the seeminglyinfinite variety of them perceived by our sensesis in reality the result of the arrangementof a relatively small number of basic units, the molecules.Moreover, analysishas
shown that only a very small numberof chemicalelementsmake up even
the most complexmolecules;that combinationsof less than one hundred
elements,in differentproportionsand relationships,
account for tens of
thousandsof compoundsand many billionsof entities.And further analysis
hasrevealedthat elementsthemselvesrepresentdifferentdynamic arrangements of only n fsw-nscording to some hypotheses,only two-fundamental corpuscles.
Upon close analysis,nature, which appearsto be so greatly varied,
turnsout, in fact, to be basedupon only a very few fundamentalconstituents and it is the manner in which theseconstituentsare bound together,
which providesvariety.
theirorganizationinto a multitudeof combinations,
I
R E S E A R c Hr N p H y s t o p A T H o L o c y
The studyof organizationobviously,then,could furnish the most valuable information about nature.And it would not appearto be too much to
expect that, if nature's seerninglyinfinite variety stems from organization
of only a few constituents,then organizationitself might also be achieved
through a few, relatively simple fundamentalpatterns.If so, seekingout
suchpatterns-systematicanalysisof organizationcomparableto the efforts
to systematizeconstituents----could
be of primary importance to better
understandingof a host of problems.
Homotropy and Heterotropy in Nature
As man attemptedto recognizeordcr in the constantchangessecn in
nature,he noted certain patternsthat appearedto indicatedefinite"laws of
nature."Someof theselaws have been observedto operateunder such a
wide variety of circumstances
that they have come to be acceptedas
"fundamentalIaws."
ln 1824, Sadi Carnot formulatedone which is known as the Second
Law of Thermodynamics.Carnot observedthat, in a given system,work
involving the transfornrationof thermal into mechanicalenergy is only
accomplishedas heat drops from high to low temperature.In more general
terms,this meansthat work accomplished
in an isolatedsystemresultsin
progressively
eliminatingdifferencesin temperature.Clausiusrecognized
this as a fundamentalprinciple and postulatedthat the amount of energy
availablefor work alwaystendstoward a maximum. This condition, called
"maximum entropy," correspondsto uniformization of temperatureand
also to homogeneousdisorganization.At first, it appearedthat the principle was in conflict with the First Law of Thermodynamicswhich expresses
the rule of conservationof energy.However, Helmholtz soon was able to
demonstrateits validity by showingthat only the secondlaw could reconcile
the first with the impossibilityof perpetualmotion.
In a more philosophicalvein, we considered,in our research,that this
Second Law of Thermodynamicsin its broadest sense could define a
fundamentaltrend toward annihilation of any existing differencesin nature, through the triumph of total uniformity. SinceClausiusused the term
"entropy" in applying Carnot's original observationto closed mechanical
systems,it has seemedpreferableto avoid confusion by utilizing another
term for this general tendency toward uniformity in its broadest sense.
Therefore, we have chosen the term "homotropy."
Despitethe theoreticallyrapid trend in the directionof absoluteuniformity, or homotropy, no such final state has yet been achieved.It must
be concluded,therefore,that someotherfactoropposedto that trend exists.
R E S E A R c Hr N p H y s r o p A T H o L o c y
electrons would come closer and closer to the nucleus and would, as
already pointed out, finally be absorbedby it. The quantum theory of
fields accountsfor the absenceof radiation and for electronsremainingin
their particular orbits. However, the concept of stationary statesfails to
explainall the propertiesof the atom, particularlyits chemicalreactivity,by
virtue of which different atoms combine to form molecules.
According to anothertenetof the quantum theory, the Pauli Exclusion
Principle,an orbit cannot be occupiedby an indefinitenumber of electrons
but, at most, by two electronsthat spin in oppositedirections.The orbits
are arrangedin shells,each shell having a definite level of energy.A shell
is completewhen it containsthe maximum number of electronscompatible
with the Pauli Principle.Completeshellsconsistof.2, 8, 18, etc., electrons.
When an inner shell has its quota of elcctrons,additional electronsmust
occupy an outer shell. Consequently,
insteadof falling into the nucleus,
the electronsin their lowest energy stateswill continue to revolve at it
considerabledistancefrom the nucleus.
As already indicated,if there were only electrostaticforces, the electronswould have long sincefalleninto their nuclei,neutralizingall electric
charges.The universewould be in a state of maximum homotropy.No
strong atomic forces would exist and no chemical reactions would take
place.The intervcntionof quantumforcesavoidsthis. It is apparent,then,
that the organizationof the atom resultsfrom the operation of two types
of forces,electrostaticand quantum, the electrostaticservingto bring and
keep nucleusand clectronstogetherto constitutethe atom, the quantum
accountingfor a motion of electronswhich preventstheir total annihilation and the neutralizationof all electricalcharges.
Homotropic and Heterotropic Forces in the Atom
We rnay now attempt to considerclectrostaticand quantum forces in
the atom in terms of homotropicand heterotropictrends.Let us hypothesize
an atomic systemin which only electrostaticforces are active and compare
it with a real systemwhich also has active quantum forces.Whereasthe
fictitioussystemwill rapidlyevolvetowardsa stateof maximum homotropy,
with annihilation of all charges,this will not occur in the real system.
When the two systemshave reachedfinal statesof equilibrium, the homotropy of the imaginary systemwill be greaterthan that of the real system.
If the quantum forces that keep the electronsaway from the nucleus in
the real atom could be withdrawn, the electrostaticforces acting alone
would bring about a state of completeannihilation,thus making available
a certain amount of energy that had previously been preservedby the
B^sIc coNcEprs
oF oRcANrzATIoN
.'
R E s E A R c HI N p H y s t o p A T H o L o c y
tum forces of the atom. Two atoms which do not have sufrcient electrons
in their external shells to complete the external shells of both can fulfill
their quantum forces by sharing some of their electrons.By achieving a
complete external shell for each atom, the sharing processsatisfiesthe
quantum forcesof both atoms.This methodof quantum fulfil.lmentthrough
the sharing of electrons also can lead to the appearanceof electrostatic
forces.If the two atoms are identical,the sharedelectronshave an intermediate position and, therefore,do not influence them. As a result, the
atoms have their quantum forces fulfilled without inducing new electrostatic forces. This is the so-called"homopolar bond." If, however, two
atoms are dissimilar energetically,their shared electrons will be located
closerto one atom than to the other, the distancebeing determinedby the
competitiveinfluenceexertedby the atoms upon the sharedelectrons.At
the same time, other electronswill be influencedby the bond, and, as a
result,their orbits will be alteredto someextent.Weakerelectrostaticforces
will result and the bond will be intermediarybetweenthe ionic and the
homopolar.Both kinds of fulfillment of quantum forces-one achievedby
transfer,the other by sharing-thus lead to the appearanceof new electrostatic forces in the ions or ionoids.
We must repeathere for emphasisthat the fulfillment of quantum forces
can take place through variousavenues,either by loss or gain of elcctrons,
or by sharingwhich can range from ionic to homopolar. The plurality of
possibilitiesfor fulfillment of quantum forces is very important, making
it necessaryto considerthe resultsof such fulfillment on a statisticalbasis.
The electrostaticforces act betweenchargedions of oppositesigns,or
betweenatoms bound by sharedelectrons.Through the balanccof thesc
electrostaticforces,bound atoms appearand correspondto ncutral formations, having their electrostaticforces fulfilled. However, it is only with
the intcrvcntionof suitablequantum forcesthat the bound atoms can form
a new entity, the molecule.
Quantum and Electrostatic Forces in Molecules
Alternate operation of electrostaticand quantum forces leads to the
organizationof atomsinto molecules.The quantum forcesin the molecules
interveneto permit organizationof theseentitiesso that the constituents
are maintainedat proper distancesand positions.The result is electrostatic neutrality.The appearanceof new quantum forces that maintain the
constituents,through their organizedmovement,at certain distancesand in
but also the stabilityof
certainpositions,insuresnot only the establishment
the new formations.Besidesvibrationalmovements,other more definite
B A S I C C O N C E P T SO F O R C A N I Z A T I O N
REsEARcH rN pHysropATHoLocy
tive charge,dependingupon the nature of the lost or gained entity, and this
is the outcome of the fulfi.llmentof the molecularquantum forces.This is
illustrated by the following examples concerning the benzene molecule,
and the carboxyl and hydroxonium radicals,in which we shall limit ourselvesto changesproducedby quantum and electrostaticforces.
In the benzenemolecule,which is electrostaticallyneutral, the electrostatic positive and negativeforces of the constituentatoms are balanced,
However,all the electronsare not in fixed positions.The ,,' electronsof the
doublebondsmove around in the molecule.Becausethe moleculeis closed,
this movementis circular, thus accountingfor the stability of the molccule,
recognizedin part by the equal reactivityof all its carbon atoms which is
encounteredunder certain conditions and results in the Kekulian forms.
The fulfillment of the quantum forces accountsfor a kind of relative
fixation of these wandering,,,electronswhich is responsiblefor the other
structuresof the benzenemoleculedifferent from the Kekulian ones. It is
this localizationof electronswith the capacityto entcr into further reactions
which results in the activation of the molecule as seen in the resulting
Dewar structureswhich in turn accountsfor active centerssuch as ortho,
meta, and para positions.These excited molecules,electrostaticallyactive,
can readily take part in chemicalreactions.Study of the mobile zrelectrons
in many other moleculesallows us to undcrstandtheir role in providing
molecularstability, while their relative localizationfavors the appearancc
of electrostaticallyactive centersin the molecule and resulting reactivity.
Here again, localizationof electronsopens up many possible avenuesto
activation.
The carboxyl and hydroxonium ions represent typical examples of
another kind of activation. Inactive carboxyl occurs when the quantum
forcescausethe electronsto wander continouslybetweenthe two oxygens
of carboxyl.Becauseof this electroniccondition,the H atom seemsno
longer to be bound to either of the o atoms,but is situatedbetweenboth:
this form correspondsto the electrostaticatly
fulfilled condition. With fulfillment of quantum forces,the wanderingelectrontakes a more fixed position at one or the other oxygen.when this occurs.the H+ ion leavcsthe
carboxyl group, and the carboxyl acquiresa negativeelectrostaticequilibrium, leadingto further combiningactivity.This fulfillment of the quantum
forcesis responsiblenot only for the appearanceof an activatedgroup of
electrostaticcharacter,but also for the existenceof two structures.each
one with anotheractive oxygen.
A similar activation takes place when a molecule acquiresan ion, as
seen for the hydroxoniumion. Water can, under ccrtain circumstances,
l0
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l l
12
xEsEARcH rN pHysropATHoLocy
13
PrincipalParts
First [ntities
Secondany
Parts
o f t h e S e c o nEdn t i t y
S e c o nEdn t i t t e s
T h i r dE n t i t i e s
F o u r t hE n t i t y
f t h e T h i r dE n t i t y
f t h e F o u r t hE n i i t y
Secondaryparts have important characteristicsin common. These become especiallyevident when lower level entitiesare examined.At levels
below the atom, secondarypartsfor all entitiesare electrons.It is of interest
to observethat in higher entitiesas well, such as moleculesor micelles,all
secondary parts have a negativeelectrical charge.
L4
xESEARcHIN pHyslopATHoLocy
15
these entities are assumedto have appeared.we will see below how important this is for the more complexentitiesof higher levels.
In the role played by secondaryparts in progressiveorganization,the
changesin their mobility are of specialsignificance.we have already seen
that in hierarchicentitiesthe secondaryparts are simpler units than the
principalparts,a factor which facilitatestheir mobility. The mobility can
be related to the fact that these secondaryparts are derived from the
environmentwhere they arc mobile, with their motion not systematized.
The interventionof quantumand quantum-likeforces,which help to create
new entities,can be seenas a kind of organizationof the relativemobility
of the secondaryparts. It is the systematization
of their movementwhich
preventscompleteannihilationof electrostaticforces present.The relationship between secondaryparts and environmentthus explains the
characterof the mobility encounteredthroughouthierarchicorganization.
It must be emphasizedhere that, becauseof the electrostaticnature of
the bond betrveenprincipal part and secondaryparts, a principal part is
capableof entering into the formation of more than one specifictype of
entity. similarly, fulfillmentof quantumforcescan lead to more than one
typc of structure,However, of the many possiblenew entities,or structures,only a few will fulfill the requirements
for developinga still higher
organization,i.e. will be capablcof acting as principalpart in a new cntity. some rcmain at their original level without progressing,
even after
being bound to other entities.Even many of thosewhich have somecapability for higher organizationcan go only one or two steps.Only a very
few will continueall the way up. [n other terms,only a few will be able
to utilizc new quantumforcesin order to realizencw entities.While various bondsand structuresoffer a largevarietyof possiblenew entities,it is
thc entity with a capacityto adcquatelyresistthe cffectsof the changing
environmentwhich will take part in progressiveorganization.
In considering the forces which intervene in progressivehierarchic
organization,one has to considerthe free cnergyavailablein the environment. The immenseamount of energyreceivedfrom the sun representsa
typc of heterotropicenergy which can intervenein organization.We will
see that this is easilyrecognizedfor higherentities.The ability of certain
entitiesto develop may be relatedto their peculiarability to utilize heterotropic forces, most of them of solar origin. The lesssuccessfuldisappearor
remain at lower levels.
16
xESEARcHIN PHYSIoPATHoLocY
CHAPTER
BIOLOCICAL ENTITIES
\xr
18
R E S E A R c Hr N p H y s I o p A T H o L o c y
BroLocrcAL ENTtTIES
19
Entities.
AddedSecondary
Format
ions
Chromomeres
Chromonemata
Fibrillar material
Chromosomial
sap
Chromosomes
lfucleus
CelI
Nuclearsap
Cytoplasma
; : : i '
. r ' :
I ]!a:; trr:4trr:itl'riitir{)
Nl
20
RESEARcH rN pHysropATHoLocy
more, organs grouped together, with blood as secondary part, form the
entity called the organism.(Fig. 3,)
Does the samepattern apply for entitieshierarchicallyinferior to chromomeres?For these lower entities, morphologicalinformation to define
the relationshipbetweenprincipal and secondaryparts is unavailablefor
Entitv
Added
Secondary
Formations
_ _ I n t e r s t i t i a lf o r m a t i o n s
Tissue--Lymphat
ic fornations
0rgans
0rganism
- -8lood f ormat
ions
Ftc. 3. The oganization abot'e the cells. The hierarchic pattern of the organization
in general is recognized also in the organization above the cells. Tissues, organs.
organism are seento representhierarchic entities,each one being made by a principal
p a r t f o r m e d b y t h e g r o u p i n go f e n t i t i e sh i e r a r c h i c a l l yi m m e d i a t e l yi n f e r i o r t o i t . T h e
principal part is bound to a secondarypart characteristicfor each entity. This secondary part would correspond to the proper environment in which the entities forming the principal part, have evolved.
BroLocrcAL ENTITtES
2l
22
nESEARcHrN PHYSToPATHoLocY
Princtpal
Added
Secondarv
Format
ions
il-C-l{-CqrouD'
Alkaline-ami!o
Aminoacid group
Histones
aminoacids
Deoxycibo-, \
nutleoproteins
Deoxyribonucleic
acld
Genes
Nucleooroteins
and
o t h e ls^e c .e n t i t i e s
B t o L o c r c A L E N T T T T E S/
23
constituents
forming the principal part for a new entity, the predominance
of one or another constituentwill make entities at the same level differ.
This mechanismof differentiationthrough the constitutionof the principal
part has been extremelyimportant throughoutthe biological realm.
Nucleolus
The concept that plural groupingscan enter into the principal part of
the nucleusputs the role of the nucleolusin a new light. It was accepted
for a long time that the nucleolus representsonly the reserve material
necessaryfor metabolism of the nucleus. The strong positive electrical
characterof the nucleus,as recognizedthrough its rather alkaline reaction,
would give it roles more important than that of the other constituents.
According to a work hypothesiswhich we advance,successivenuclcolar
formationswould representthe principal parts of hierarchic organization
below the nucleuslevel. In chromonemata,chromosomeand nucleus,the
partscorrespondingto the nucleoluscan be recognized.These formations
are grouped together with genesto form the principal part of the chromonemata.Similarly, chromatineformationsrepresentingentitiesof the
samelevel as the respectivenucleolar formations,will form togcther the
groupscharacterizingthe principal part of chromosomes.In the nucleus,
the nucleolusis joining the other formationsto form is principal part.
Protop lasmatic F ormations
In the cell, a hierarchicallysuperiorentity, a similar condition also
appearsto persist.The protoplasmaticformationswith ribo-nucleicacid
can be conceivedas representingentitiesof a nuclearlevel, that is, a level
similar to that of the nucleus.Togetherwith the nucleusthey would form
the group correspondingto the cell. This kind of evolution of entities in
relatively separateparallel lines, with their further grouping together to
form principal parts for new entities,is part of the typical pattern of
organizationespeciallyevidentin the biologicalrealm.
Boundary Formalions
We have mentionedthat groupingsof severalentitieswould not be
sufficientto form a new entity so long as the secondaryenvironmentalpart
is not isolatedfrom the medium from which it originates.Consequently,
the new entity appearsonly when a distinctboundaryformation is formed.
Progressivehierarchic developmentis dependentupon the appearanceof
suchboundary formations.For the first biologicalentities,the radicals,the
boundary semsto be more an energeticproperty than a morphologically
24
nESEARcH rN PHYStoPATHoLooY
BroLocrcALENTrrrEs
25
Enti ty
ChromomeresChromonemala/-Chromosomes.Nucleus- -
formation
Boundary
boundary
Chromomere
boundary
Chromonemata
membranE
Chromosomial
-- Nuclearmembrane
-.Cellularmembrane
- Lymphat
ium
ic endothel
C eIIs
Tissues-J-
0roans
- F-- I
\
B l o o dv e s s e l s
S k i na n dn u c o u s
menbranes
\
\
\
0rganism
26
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BfoLoorcAL ENTtTTES /
27
28
REsEARcH rN pHysropATHoLocy
BroLocrcAL
ENTtTTES
29
30
nESEARcH rN pHystopATHoLocy
|later Circulation
The passageof complex organizedanimals into the new environment
of terrestrial life brings to the fore the problem of the place of water in
hierarchic systematization.It appears to us an acceptable concept that
water doesnot circulatefreely in the organism.Its appearancein hierarchic
entitiescan be understoodif, as we did for the other constituents,we relate
water to its place in the environmentin which phylogeneticdevelopment
has taken place, In the first near-volcanoenvironments,which applied to
the subnuclearentities,water was relativelyscarce,which explainsthe high
concentrationof the constituentsin the nucleus.The mud of the earth's
crust is richer in water, which explainsthe difterencebetweenthe nucleus
and the cytoplasm, with the latter richer in water. The sea was the
environmentfor the metazoiccompartment,which explainsthe richnessof
water in this compartment.The so-called"internal sea" consequentlycan
be seen only in the metazoic compartment.With the passageinto the
terrestrialenvironmentwith its air medium,the water againbecomesscarce
and has to be conserved.The circulationof water betweencompartments
is governedby osmotic forces which are determinedby the original richness in water of the respectiveenvironments.The importance of water
circulation appearsevidentwhen an abnormalityin its distribution occurs.
Water arriving in a compartment-alone or with a cation-in an amount
abovethatcorresponding
tothe constantfor that compartment,isseparated
from the constituentsof the entity in order to reestablishthe characteristic
constant value. Such separationof abnormal amounts is accomplished
accordingto the compartment,through the appearanceof vacuoles,edema,
exudatesor diuresis.
Animals and Plants
The conceptof hierarchicorganizationin which each entity canconserve
its own environment allows us to consider in a new light various other
problems of living organisms.One concernsthe fundamentaldifferences
betweenanimalsand plants.Analysisof the constituentsof the metazoic
secondarypart providesa new criterion for distinguishingbetweenanimals
and plants and gives logical meaningto its distinction.Animals can be
characterized
as having sodium as the cation of thcir metazoiccompartment; from the cell level on, they have had the sea as their temporary or
even permanentenvironment.Plants,on the other hand, have potassium
as the principal cation for their metazoic compartment, indicating that,
from the cell level on, they have had the earth'scrust as their environment,
B r o L o c r c A LE N T r r r p s /
lt
32
REsEARctf IN
PHYSIoPATHoLocY
B I O L O c T C A LE N T I T t E S
33
sistenceof constantsproper to each entity is distinctly opposed to homotropy. Life in its broadestsense,correspondsto the capacity of an entity
to maintain heterotropyby conservingits characteristicconstants.The life
of any entity aPpearsto be synonymouswith conservationof its constants.
An entity dies when it has permanently-that is, irremediably-lost its
capacityto conservethe constantswhich characterizeit. Death then representsexhaustionof heterotropyfor the specificentity.
The fact that, in essence,life appearsto be synonymouswith the conservation of constants and is heterotropic,relates it, and especiallyits
origin, to one of the important sourcesof heterotropicforce, solar energy.
A distinction has to be made betwcen heterotropyas one of the fundamental laws of nature and the meansby which it is exerted.Solar energy,
with all of its quantas,would greatly increasethe effectsof heterotropic
forces in nature. It would not createsuch forcesbut would simplify them
and extend their applications.The origin of matter and, as we have seen
above, of entities, biological or nonbiological,is in the final analysisthe
result of heterotropicforces.External conditionsqualitatively and quantitatively influenceoperation of heterotropicforces.
The sun's heterotropiccontributionshave to be consideredunder this
aspect.Throughthe quantasit disposes
of, solarenergyhasnot createdlife,
as conceivedabove, but by permitting more and more entities to appear,
has greatly facilitated their extension.Its effect, although certainly not
limited to any group of entities,seemsto be especiallyimportant to those
forming the biological realm. Similarly, the effect, of a special type of
energ.y,radiation,also must be considered.Radiation appearsto be related
to the elements,and will be discussedin a later chapter devoted to the
elements.
Sincelife itself is related to changesdirectly aimed at conservingconstants,in this broadestsenseit is no longer limited to the specificgroup of
cntitiesfound in the "biological" realm. Life has the samemeaningfor an
atom, crystal or micelle, as for a cell, organ or organism.It is for this
reason that knowledge of the mechanismthrough which constancy is
achievedbecomesof great importancein the study of all matter and especially of the biologicalrealm.
Maintenattce ol Constants
To study the mechanismused to maintain constants,we must define
exactlywhat constancymeans.According to Cannon'sprinciple of homeostasis,
constantshave beenconsidered
to correspondto a dynamicbalance
that results from the continuousoperation of two opposingfactors. And
34
R E S E A R c Hr N p H y s r o p A T H o L o c y
BIOLOCIC^L ENTITIES
35
36
nEsEARcH rN pHysIopATHoLocy
BtoLOctc^L ENTTTTES /
37
CHAPTER
CANCERAS AN ORGANIZED
CONDITION
il
CANCER ^S
^N
ORGANIZEDCONDITION
39
40
REsEARcHrN pHysrop^THoLocy
tion which is one of the basic characteristicof the epithelium. Since the
regularrelationshipbetweenthe cells forming epitheliumcan be attributed
to dipolarity, the anarchicdispositionseenin this phaseof cancer can be
ascribedto loss of cellular dipolarity.
cancer-in-situ, in terms of hierarchic organization,would appear to
involve the level of the nuclei, and the noninvasiveness.
characteristicof
this phase,persistsas long as the "cancerous"abnormatityremainslimited
to this level, that is, as long as the cytoplasmof cells remains apparently
unaffected.
Invasive Phose
This phaseis characterizedby irregularproliferationof cells and penetration into neighboringtissues.To the anarchicarrangementsnoted in the
noninvasivephase, now has been added exaggeratedgrowth. And the
changeof a noninvasivecancer into an invasiveone can be consideredto
result solely from the addition of the new factor of abnormal growth. The
invadingcells will persistonly if, concurrently,there is a loss of the defense
mechanismof the invaded tissues,as will be seen later.
Studiesof invading cells have revealed,in this phase,an anomaly no
longer limited to the nucleusbut now encompassing
the cytoplasmas well.
Exfoliative cytology has shown an abnormal and rapidly disintegrating
cytoplasmand this has servedas an important diagnosticcriterion. From
the point of view of organization,it can be said that, with the participation
of the cytoplasm,the diseasehas progressedfrom the nuclear to the cellular level.
Painlul Phase
Pain is the principal clinical manifestationcharacterizingthe next phase
of the disease.As we shall explain in greaterdetail later, pain arisesfrom
changesin the pH of the intercellularfluid that bathessensorialnerve endings.For the moment, we can remark that biochemicalchangesnow occur
outsidethe cells, and, with the participationof interstitial formations,the
diseasehas progressedto the tissularlevel.
Preterminal and Terminal Phases
In the next stage,the preterminal,biochemicalchangesaffect the function of variousorganswhich may or may not in themselvescontain cancerous cells. while some changesin function may be seen even before this
preterminalphase,now, there is manifestimpairment.And, while the invasionof an organ by cancerousmassesis a factor precipitatingthe func-
c A N c E R A S A N
O R C A N I Z E DC O N D T T I O N
4 l
tionalchanges,
invasionis not indispensable.
Abnormatbiochemicalchanges
leadingto seriousfunctionalimpairmentsare seenin organsentirelyexempt
from tumor masses.
with further progress of cancer, metabolic functions that are systemicallyimportant becomeabnormal,Later, we will analyzr,in detail these
changeswhich affect the whole organismprofoundly.For the moment we
want only to note that, with thesechangescancerpassesfrom the clinically
preterminal to the terminal phose.
In the light of this systematization,cancer then appears to progress
clinically in organized fashion as it passesfrom the relatively innocuous
nuclearnoninvasivecancer-in-situto a lethal systemicdisease,the progress
being marked by the successiveparticipationof difterent hierarchic levels
of the organization. Table I sums this up.
T,fSLP
Organizational
Level
Subnuclear
Nuclear
Cellular
Tissular
Organic
Systemic
PhysiopathologicalChanges
Gene and chromosomeanomalies
Nuclearanomaliesand atypical
cellular arrangemenls
Atypical growth
Local pH changes
Organic metabolicchanges
Systemicmetabolicchanges
Ctinical phase
precancerous
Noninvasivecancer
Invasivecancer
Painful cancer
Preterminalcancer
Terminal cancer
42
RESEARcH rN pHysropATHoLocy
CHAPTER
DUALISIU
'fhe
/ \\s wE HAVEotssERvED
ABOVE,
dualismprevailsin nature.
concept
of an oscillatorydynamic balance-the result of alternateopcration of opposedforccs-has beenof specialvalue in the study of most of the physiological phenomcna.Over the yearswe have alsoconstantlyobservedthat in
most physiopathological
manifestations,
dualisticpatternscan be recognized.
This dualisticpathogenicconccpthas helpcd to guide our study of disease.
In cancer, it has permitted bettcr understandingof many processesand
manifestations.It has also servedas a basis for our attemptsto influence
cancer and other conditionstherapeutically.
It was in the study of pain
that, initially, we found clear evidenceof pathogcnicdualism.
PAIN
Many years ago, during expcrimentswith an alcoholic cxtract of human placentaas a therapeuticagentin terminalcancercases,a curiouseffect was observed.In somepatientswith painful lesions,administrationof
the preparationresultedin a decreasein the intensityof pain and even in its
disappearance
within a few minutes,with rclicf usuallylastingfor hours. In
other cases,however,there was an oppositeeffect;pain increasedin intensity within a few minutesafter an injcction.In somesubjects,the exacerbation was so great and pain becamcso unbearablcthat the experimental
treatmenthad to be discontinuedquickly. In severalcasesin which the
preparationwas used in progressivelylarger doscs,another notcworthy effect was observed: after the first injections,pain decreasedand even disappearedfor severaldays, only to have a new pain arise as treatment
continued.This new pain becamemore intcnseaftcr each injcctionso that
the therapyhad to be discontinued.
Patientsclearly recognizedthe differpains.
encebetween
The new one frequentlyhad a burningcharacter.
43
44
REsEARcH rN pHysropATHoLocy
DUALTSM
45
46
nEsEARcH rN pHyslopATHoLocy
DUALTSM
47
pH
8.O
Unne
7.5
7.O
6.5
6.O
5.5
5.O
to
tz
'6
,.1,!
tB
22
24
48
RESEARCH IN
PHYSIOPATHOLOGY
PAIN
Unbcorobh
Vcry Scvcn
Scwrc
Moduote
S,,9ht
l{ona
0H
Utina
6.2
6.O
58
5.6
5
5.2
5.
l'lour
Fto. 7. The alkaline paltern ol pain in which the concomitant variations in the curves
of pain intensity and urinary pH are parallel, seen iD a case of arthritis.
pH
7.5
Uttna
7-O
6.5
6.O
5.5
5.O
Hour
Ftc. 8. An ocid pain patlern is seenin a caseof carcinoma of the prostate with meta.
s t a t i c b o n e l e s i o n s ,i n w h i c h t h e c o n c o m i t a n tv a r i a t i o n so f t h e c u r v e s o f p a i n i n t e n 5ity and urinary pH are divergent.
DU^LrsM
49
_N_
Utlte
8.O
7.5
7.O
6.5
6.O
5.5
llou I
Ftc. 9. An acid pattern ol pain recognizedby the divergent variations of the curves
o f p a i n i n t e n s i t y a n d u r i n a r y p H i s s e e ni n a c a s eo f p h a n t o m l i m b .
50
nESEARcHrN pHysropATHoLocy
C o r c r n o m oo f R e c t u m
|9 4 3
+++ +
+ + +
c
++
o
o+
8.2
74
--
6'2
54
46
C o u r t e s yo f D r .M o r r oR o g n o n i
Fto. 10. The pain pattern can be recognizedalso through the characteristicopposite
variations of the curves of pain intensity and urinary pH, followed during successive
days-instead of hours-as seen in the above curves of a case of cancer of the rectum. (Courtesy of Dr. Rognoni)
operative and accidental wounds, burns and fractures, for example-always has an alkaline pattern. This is also true for the pain of gallbladder
colic. In other conditions,eitherpatternmay be presentand must be determined by analysis,For instance,the pain of neuritis and simple headachehas an acid pattern in somecases,alkalinein others.
In rheumatoidarthritis,an alkalinepain is almost constantlyfound.
ln osteoarthritis,the pain is of an acid type. In arthritic patientsin whom
DUALISM
5I
Pain
i n t e n ist y
6
4
2
(a)
K+ in serum
in mEq
.2
.0
1
H o u r s
Frc. I l. Pain pailern and potassium in blood serum. The comparison of the concomitant changesin the curves of pain intensity and those of the amount of potassium
in blood serum shows parallel variations in a case with an alkaline pattern.
52
RESEARCH IN
PHYSIOPATHOLOGY
t0
Pain
Intensity
1
7
o
(b)
1.0
K+ tn serun 58
i n dq
16
54
32
3
Hours
F t o . 1 2 . D i v e r g e n t v a r i a t i o n sb e t w e e np a i n i n t e n s i t yc h a n g e sa n d t h o s e o f t h e b l o o d
serum potassium in a case of acid pain pattern.
PAIN LESION A
YaTt
Sevlrl
'.odarota
Sllghl
Nonr
MIN LESION8
Vr"t S.v.r.'l
Stvrrr
I
Hod.rlt
I
srbhf
{
I
ttolr
n!__UntlE ?.o
6.5
60
3.5
Frc. 13. Pains. Acid and alkaline pains can co-exist on different lesions,as seen in a
patient with multiple osseous metastasesfrom breast carcinoma. Lesion A, which
c o r r e s p o n d st o a n a c i d p a t t e r n . s h o w s d i v e r g e n t v a r i a t i o n s b e t w e e n t h e c u r v e o f
u r i n a r y p H a n d t h a t o f i t s p a i n i n t e n s i t y ,w h i l e f o r l e s i o n B . w i t h a n a l k a l i n e p a t t e r n ,
t h e v a r i a t i o n so f t h e c u r v e so f p a i n i n t e n s i t ya n d o f u r i n a r y p H a r e p a r a l l e l .
I _H..1"*_
:" .
_,
l,lil{Si"Ss\jilJ,1,$s-\i1,,i"r".+r,,"
DU^LrsM /
53
HrPOr
7.O
6 .o
5 0
4.O
8
9
t
Houl
F t c . 1 4 . T h e c h a n g e si n d u c e d i n t h e p a i n i n t e n s i t yb y t h e a d m i n i s t r a t i o no f a n a c i d i fying agent indicate the pattern present. Pain with an acid pauern is intensified foll o w i n go r a l a d m i n i s t r a t i o no f 1 . 5 c c . o f . a 5 o % s o l . o f p h o s p h o r i ca c i d . U r i n a r y p H
changesreflect the induced systemic acidification.
changes
occur rapidly. In a caseof sciatica.we have seenrapid and frequentchangesin pattern,especiallyin responseto therapeuticmeasures.
Worthy of being noted is the correlationfound betweenvariationsin
painintensityand changesin the acid-basebalanceof the body even in
casesin which nerves are directly involved in lesionsand in which a
mechanicalpathogenesis
usuallyhas been accepted.This would indicate
thatthe chemicalfactor mentionedabovehas a role in the pathogenesis
of
paineven in thesecases.
Acidilying and Alkalizing Agents
We next demonstratedthe cause-effect
correlationbetweenacid-base
changes
and variationsin the intensityof pain. Administrationof acidifying or alkalizing substancescould induce the same changesin pain
intensityas those causedby spontaneous
variationsin the acid-basebalanceof the body.
The relationshipbetweenacid-basebalancechangcsand pain intensity
was thus investigatedby administeringstrong acidifying and alkalizing
54
RESEARcH rN PHYsropATHoLocy
PAIN
Severe
Poo
JHt
Moderote
Slight
None
pH
Urine
7.O
6.5
6,O
5.5
Hours
Flc. 15. Pain with an alkaline pattern is relieved by oral administration of two doses
of 1.5 cc phosphoric acid (50Vo sol.). Urinary pH changes reflect the systemic acidification.
Sodium bicarbonate or ammonium acetate in quantities that alkalinized the urine increasedintensityof pain with an alkaline (Fig. I6) ^nd
diminishedintensityof pain with an acid pattern.(Fig. 17)
These changes of the systemic acid-basebalance induced by administration of strong acidifying or alkalizing agents explains how similar
changes,when they occur spontaneously,affect pain intensity. The effect
DUALISM
PAIN
Unbrorcble
Very Scvere
lVoHCOr
Sevcre
Moderote
S1 , 9 h ,
NonG
pH
Uttt'
/ s s
80
7 0
6 0
5.O
9
Houl
F t c . 1 6 . P a i n w i t h a n o l k a l i n ep a l t e r n i s i n t c n s i f i e db y o r a l a d n r i n i s t r a t i o no f 5 g r a m s
of sodium bicarbonate.urinary pH changesreflect the systcmic alkalinization.
fllA/
U^bP'oroble
Wy Severe
Sevcrc
Modercle
NoHCOs
Slighl
None
PH
Unne
8.O
7O
6.O
a n
I
a
Houl
F t c , 1 7 . P a i n w i t h a n a c i d p a t t e r n i s r e l i e v e dh y o r a l a d m i n i s t r a t i o no f 5 g r a m s o f
s o d i u mb i c a r b o n a t e .U r i n a r y p H c h a n g e sr e f l e c tt h e s y s t c m i ca t k a l i n i z a t i o n .
56
xESE^RcHrN pHysropATHoLocy
Hours 35
unbccroblc
vcry rcvtra
t cv0ra
modorolo
rlight
nona
Co of Prortolr
f5
25
35
45
1 . 5g m r . A m o n i u m
55
05
15
25
9m3A
. monium
occloll
ocr tot.
r.5eT
A m o n i u mo c c l o l c
Go of U l c r u r
fo
20 30 4 0
oo
oo ro
l
unbcoroblc O . 5g m t . A m o n i u mC h l o r i d r
varyr
Co of 8r.ort
20
30 40
50
l g m . A m o n i u mC h l o r l d r
taYcra
nodcroh
I light
nona
Co of Ulcrur
Co of Uteruc
30
YaTy scvarc
stvcrS
40
50
00
ro
to
? o r c 4 p 5 0
modorolc
rllght
nonc
Ftc. l8bis. The responseof the pain of a lesion to an agent permits to identify the
acid or alkaline pattern present.The effect of an acidifying and alkalizing agent correspondsto an increaseor a decreasein pain intensity, according to the pain pattern
present.First row, left side-acid pattern, right side-alkaline; second row, left sideacid, right side-alkaline; third row. right side-alkaline, left side-acid pattern.
DUALTSM
57
formed. Specialglasselectrodest were used for this purpose and determinations were carried out employing a sensitivepH meter. The tip of the
electrodewas placed on the surface of the area to be testedif ulcerated,or
was introduced into the tissue to be tested through a small incision. In
reality this givesa measurementof the pH of the local interstitialfluid.
Urine pH and local pH determinationswere then performed at difterent
times correspondingwith spontaneousvariationsin the pain intensityexperiencedby the patient. Simultaneously,the pH values of normal tissue
areas and, when possible,of nonpainful tumor areas were determined.
Similar studieswere carried out after administrationof strong acidifying
and alkalizingagents.
Many difficultieswere encounteredboth in the choice of suitableclinical subjectsand in techniques.Neoplasmsproved to be the simplesttype
of painful abnormalprocessto employ.The neoplasmhad to be locatedin
a readily accessibleregion so that the electrodescould be introducedinto
an ulceratedarea or through small incisions.The patient had to be able
to very accuratelylocalize the area of pain since considerabledifferences
in pH valueswere found to exist in difterentparts of the same lesion.The
pain had to be superficiallylocalizedbecauseaccuratedeterminationsin
the depths were not possible.Finally, the complete cooperation of the
patientwas essential.
The data obtained for a patient with an ulcerated,profusely draining
carcinomaof the breast is recordedin TnsLe II. Pain was most intense
T,rsl.e II
Onse.nvrtroNsIN A Crse wlrn Alx.,rllxe P,rlx P,rrrenx
Treatment (oral)
Phosphoricacid
S O V o . 2c c .
Sodium bicarbonate
5 grams
pH Normal
Tissues
pH Tumor
None
Moderate
Very severe
5.4
6.2
7.1
7.3
7.4
7.6
8.1
8.5
Slight
5.5
7.3
7.9
Unbearable
7.8
1 A
I .t+
8.8
58
R E S E A R c Ht N p H y s r o p A T H o L o c y
pH TUMOR
Treatment(oral)
Pain
Intensity
None
Moderate
Severe
Phosphoricacid
5 0 V o, 1 . 5 c c .
Unbearable
Sodiumbicarbonate
5 grams
None
p H r pH Normal Painful
Urine
Tissues i area
7.0 l
7.4
i 66 . 3r
Nonpainful area
5.8
5.3
t.)
5.8
7.2
7.1
6.9
5.0
t.-)
5.5
6.8
6.6
7.2
l.+
DU^LrsM
59
60
REsEARcHrN pHysropATHoLocy
Abnormal Substances
changes in the local pH, like changesin acid and alkaline patterns,
could be related to the appearanceand subsequentaccumulationof difterent substancesin the lesion. Increasedconcentrationof lactic acid was
found in the interstitialfluids of acid pattern lesions,while increasedconcentrationsof sodium and especiallypotassiumions were found in the
interstitialfluid of lesionswith an alkalinepattern.
Processesleading to a local acidosisare known and ascribed to the
well-known anoxybiotic metabolismof carbohydrates.They result from
lack of the "respiratory" oxybiotic phase of carbohydrate metabolism,
with the consequentconversionof pyruvic acid into lactic acid. only part
of the lactic acid is changedinto glycogenthrough the pasteur-Myerhoff
reaction,and an accumulationof lactic acid results.While the presenceof
lactic acid is well known, its presencehas not previously been related to
pain or other manifestations.
We could showthis correlationin somecases.
(Note 3 )
The scientificliteratureofreredno information concerningthe appearance of alkaline compounds.We were able to establishthat the presence
of sodium ions coincideswith another anomaly found in theselesions-a
high fixationof chloridesin the lesionsthemselves.
Valuesof 1600 mgr. of
cl., or even higher, per 100 gms. of wet tissuewere found in theselesions
insteadof about 400 mgr./ 100 gms. measuredin normal tissues.The local
alkalosisand the resultingalkaline pain pattern could thus be correlated
with an abnormal sodiumchloride metabolismat the tissularlevel in which
with chloride ions fixed by the cells, sodium ions remain free to combine
with carbonate anions and form alkaline compounds. If the abnormal
NaCl metabolismoccurs in the interstitial fluid, the subsequentalka.losis
inducespain. We will seelater how abnormal NaCl metabolismalso takes
place at other levelsof the organization.In thesecasesof alkaline pattern
of pain, the fact that abnormalamountsof sodium ions still enter the cells
will result in a loss by thesecells of potassiumwhich will accumulatein
the pericellularfluids,and form alkalinecompounds.
An immediateconclusionthat could be drawn from these studieswas
that there is a definitedualismin pathogenesis
of pain originatingin abnormal tissuesand that the two pain patternsevidencedin lesionswith acidosis
or alkalosisof the interstitialfluids indicatethat processesof two opposite
naturesgo on at the tissularlevel.
DU^LtsM
6l
SYMPTOMS
using the same method of investigationas in the study of pain, a relationship betweenvariationsin intensityof certain other symptomsand
variationsin urinary pH could be found. Acid and alkaline patternsof
itching could be recognized.The samepatternscould be found for vertigo
and impairedhearing.Among psychiatricmanifestations,
manicdepressive
statesshowedchangesthat could be relatedto acid-base
variations.
The correlationbetweendyspncaand acid-base
variationswas contrary
to what we expected.classically,dyspneais believedto be associated
only
with an increaseof acidityof blood. However,both acid and alkalinepatterns of dyspnea were observed,and could be related more directly to
changesat the tissuelevel.We will rcviewherethe pathogenesis
of itching,
vertigo,dyspnea,and other conditions,undcr this dualisticaspect.
Itching
The problem of itchingis interestingfrom other than the therapeuticpoint
of view, sincelittle is known regardingthe natureand causeof this condition. Various hypotheses
have been offeredbut have failed to explain its
pathogenesis.
our interestin itching originatedduring the study of pain
and led to an hypothesiswhich allowsus to approachthe problemfrom a
new angle. By analogy with pain, we separateditching into two types:
physiological,
as the responseof normal tissuesto externalstimuli; and
pathological.as a sensationarising within diseasedor damagedtissues.
Ticklingthe normalskin or evenstimulatingit throughheat.colcl,etc.,may
causa sensationof itchingand lead to scratching,Certainmucousmembranes,such as thoseof the nose,and the skin aroundnaturalorifices,arc
especially
sensitiveto such stimulation.This type of itching,as a response
of normal tissuesis phy,siological
or sensoriul.
Under pathologicalconditionshowever,the skin, mucousmembranes
andother formationsmay itch without externalstimulationor in response
to stimuli which ordinarilydo not producethis sensation.
The itching then
can be consideredas a manifestation
of diseasedor damagedtissuesand,
as such can be describedas pathological.Just as does pathologicalpain.
pathologicalitching representsa symptom rclated to abnormal changes
afreadypresent.(Note 4)
In spite of their relative independence,
the fact that similar fundamentalmechanismsare involved in the productionof itching and pain
explainscertain characteristics
they have in common. Like pathological
62
R E S E A R c Hr N p H y s I o p A T H o L o c y
lTCHING
Very Severe
Seyere
Moderote
Slight
None
PH
Urine
5.5
6.O
5.5
5.O
t
4
Hours
DUALTSM
63
sity of their itching for each hour, rather than to indicate the maximum
intensity at the exact time of recording. voided urine specimenswere
obtained at the end of each hour. The pH of the urine specimenswas determined electrometrically.A graph was plottcd to compare thc variations
in the subjectivedata furnishedby the paticntwith the concomitanthourly
changesin the pH of the urine. It was usuallynecessaryto repeat the test
severaltimes before the patient appearedable to satisfactorilyevaluatethe
changesin the intensity of the symptomfor hour-long periods rather than
ITCH IN G
VerySeyere
Severe
Moderote
Slight
IVone
fi_
Urine
7.O
6.5
6.O
5.5
Hour
FIc. 21. An acid patlern ol itching is recognizedin a caseof pruritus ani through the
divergencein the concomitant variations of the curves of itching intensity and urinary pH.
for just the moment of recording. (we also tried to judge the intensity of
itching through the frequency,intensityand duration of scratching,as
notedby an observer,but without success.)Fiftecn patientswere studied
and, becauseof the limited number, the resultsare presentedas merely
preliminary.
64
x E S E A R c Ht N p H y s l o p A T H o L o c y
In four cases,the curvesof itching and urinary pH did not show any
definitecorrelationeven after repeatedtests.of the remainingI I cases,7
showed a distinct parallelismof the two curves, Md in the other 4, an
inverserelationshipbetweencurveswas apparent.The graphs obtained in
two characteristiccasesare presentedhere. (Frgs. 20 and 2l )
Severe
lvloderole
Slf9ht
HsPOe
None
pH
Urine
7.O
6.5
6.O
5.5
5.O
Hours
F t c . 2 2 . A d m i n i s t r a t i o no f p h o s p h o r i ca c i d - 1 . 5 c c p h o s p h o r i ca c i d , s o l . 5 0 c Z - i n d u c e s t o g e t h e rw i t h a n a c i d i f i c a t i o no f t h e u r i n e , a n i n c r e a s ei n t h e i n t e n s i t vo f t h e
itching with an acid pattern.
DU^LtsM
65
\\'asmore acid, and less severewhen the urine was more alkaline.This
representsan acid pattern.
The effect of a strong acidifying agent,phosphoricacid, in caseswith
acid and alkalineitching,is illustratedin Figures22 and 23. The intensity
of the alkalineitchingin the first casewas reducedby the acidifyingaction
of phosphoricacid, while the acid itching of the secondcasewas intensified. In Figure 24, the responseof a patient with alkaline itching to the
administrationof sodium bicarbonateis shown. The intensityof itching
wasgreatly increasedafter the alkalizingagent was given.
ITCHING
Seyere
IH'Po'
Moderote
Slight
None
pH
Urine
7.O
5.5
6.O
5.5
4
Hours
F l c . 2 3 . l t c h i n g w i t h a n a l k a l i n ep a t t e r n i s r e d u c e di n i n t e n s i t yb y t h e a d m i n i s t r a t i o n
o f a n a c i d i f y i n g a g e n t . I n t h e a b o v e e x a m p l e ,t w o d o s e s ,e a c h o f t . 5 c c p h o s p h o r i c
a c i d( 5 0 % ) , w e r e n e c e s s a r iyn o r d e r t o o b t a i n t h i s e f f e c t .
66
R E S E A R c Hr N p H y s r o p A T H o L o c y
evokethe sensation
of itching.More intensepH wouldresultin pain.The
fact that itching,one of the principalsymptomsof dermatological
conditions,can be relatedto local acid and alkalinechangeswithin the skin
wouldpermit the integrationof skin pathologyin a more generalphysiopathological
mechanism.
The conceptof the interventionof two difterent
ITCHING
Seyere
lvloderote
Slight
None
pH
Urine
7.O
6.5
6,O
5.5
6
Hour
F r c . 2 4 . T h e a d m i n i s t r a t i o no f 4 g r a m s o f s o d i u m b i c a r b o n a t et o a s u b j e c tw i t h a n
alkaline patteril of. itching induces together with the alkalinization o f t h e u r i n e , e x acerbalion of the symptom.
DUALTSM
67
Vertigo
vertigo has been of specialinterest in our research.while it can be
inducedby various etiologicalfactors, a basic dualism in its pathogenesis
is evident.
Many patientsrvith vertigo have been found to experiencewide variations in the intensity of the symptom. Some have exacerbationsin the
morning,others in the evening.This observationsuggesteda possibledual
patternand we investigatedvertigo by the samemethod usedfor the study
of pathological pain.
In patients with vertigo, the intensitywas determinedat hourly intervalsby using either a scaleranging from 0 to 10, or a seriesof qualifica-
x
@
g
(u
5
U'
o
OJ
g
N
N
8.7
7.
H
!
7o
o 5.6
L
o
c
L
=
,'D
46
Noon
pm.
midnightatn.
Frc. 25. Shows the curves of the intensity of lerligo and of the urinary pH in a case
w i t h a n a l k a l i n e p a t l e r n , t h e v a r i a t i o n so f t h e t w o c u r v e s b e i n g p a r a l l e l . ( 8 . W e l t ,
A M A A r c b i v e s o f O t o l a r y n g o l o g y ,5 8 : 2 7 3 - 3 0 0 ,1 9 5 3 . )
68
REsEARcHrN pHysropATHoLocy
:\
v,
c
o
c
o
CD
L
(u
o
(_
ftt
E
L
Hours
Ftc. 26. Vertigo with an acid pottern. The symptom is more intensive when the
urinary pH values are lower. (B. Welt A M A A r c h i v e s o f O t o l a r y n g o l o g y ,5 8 : 2 7 3 300,1953.)
DUALtSM
69
500cycles
--- 1000cycles
-2000 cycles
o2o
C)
'3a
o
o
,'=u4j)
an9^-to-tl-t2-/^;-Z-J
zt
Elq
L
Eto
L
I
n.n
t2
HouFs
t
e^1.
Fto. 27. The analysis of the concomitant changesin thc curvc of the intensity of thc
auditive acuity and of the urinary pH shows opposite variations iu a subject with
impairedhearing. The opposite variations arc especiallymanifest for some velucr2000cycles in this case. This rclationship corrcspondsto an acid pottern. (Courtesy
of Dr. B. Welt.)
70
*EsEARcH rN pHysropATHoLocy
:b
.at
1to
{A
tO
&
.t-
r-.
a6
a-
F. f{.
In i t i a l
PostPlacebo
P o s tA c i d i f i c a t l o n
u,
q,
o
o
o,
C
.n
C'
-t
ttiohtear A.C.
\.
\
F r c . 2 8 . T h e a d m i n i s t r a t i o no f a n a c i d i f y i n g a g c n t - 2 g r a m s o f m o n o a m m o n i u m
phosphate-induces a marked increaseof the heoring acuity if. the abnormality presenl correspondsto an alkaline pattern. Only minimal or no changesare seen to occur
if a few drops of acetic acid are used as placebo. (Courtesy of Dr. B. Welt.)
7l
F. |l.
- - - lnltial
"'-" PostPlacebo
-Post Acidification
o
o,
D
(,
c,
o
c
ti
an
C'
J
L e f t e a rA . C .
.*i*r#d'ltW$l$*iliffik,s;l.ii$i,,i#:
72
RESEARCH IN
PHYSIOPATHOLOGY
o
A. l.
--- Initral
F o s tA c i d r f r c l i r o n
o
tf,
20
zf
?t
(y
cl
IJ
&
,t
o,
c ,o
rt
an
o
af
to
Left car
to
DUALISM
73
Impaired Hearing
In studying thc various otologicalprobrems,B. welt and I noted
thatmany patientswith impaired hearingexperiencedvariationsin auditive
acuityat different times of the day, or even in conjunction with food intake.This led us to investigateimpairedhearingby the samemethod used
for pain. (Fis. 27) we studied the influenceof acidifying and alkalizing
agentsupon auditive acuity in casesof impaired hearing.
complete audiograms were obtained, employing differences of only
two decibelsbetweenmeasurements
and using all of the acceptedfrequenciesfor air and bone conduction.Audiograms were obtained in subjects
beforeand after administrationof acidifying agents,such as ammonium
fe
7fo
t6o
K.H.
- -- lnitial
' ..' ....
PostPlacebo
P o s tA c i d i f i c a t i c n
o
o
D
(J
c,
o
c
t^
o
o
'at-e'a
-'-
-./'/-./u"
'"
a-.
L e f t E a rA . C .
F r c . 3 t . T h e a d m i n i s t r a t i o n o f 2 g r a m s o f m o n o a m m o n i u m p h o s p h a t ei n d u c e s a
decreascin hearing acuity if the abnormal pattern is acid.
74
R E s E A R c Hr N p H y s r o p A r H o L o c y
K .H .
----|nitial
- * - - P o s tP l a c e b o
-f65[
Acidification
U'
o,
-r/--t'
II
-'1
I
(u
o
t
"t""'-
o
U'
o
llight Ear8 . C .
ta
2ooo
4ooo
K.H.
at
o
o
--- lnitial
' - - - - PostPlacebo
-Post Acidification
(,
o
ct
C
v,
U'
C'
-J
L e f t E a r8 . C .
Ftc. 32. Acid pattern of hearingimpairment.
was obtained with one group of agents,an opposite effect was obtained
when an agentof the oppositegroup was administered.
changes of at least 6 decibelswere required before they were considered to be induced by an agent.when changesof such intensity were obtainedwith an agent,it was invariablytrue that oppositechangeswould be
obtainedwith an oppositeagent.It was also true that the same responses
DUALTSM
75
Manic-depressiveCondition
We have studiedthe relationshipbetweenintensityof manifestations
and
systemicacid-basebalanceof the body in another group of subjectswith
variousmental disorders.of all the casesstudied, the only condition in
whicha clear relationshipcould be shown was in manic-depressive
subjects.From patients presentingchangesduring the day, passing from
periodsof high excitation to calm, from deep depressionto calm, we obtainedhourly urine specimens.At the same time, observationsof their
mentalstate were made. The evaluationsof mental condition were made
by trained observersor membersof the family using a conventionalscale
whichpermitted translationinto graph. The pH of the urine sampleswas
measured
electrometrically.Curvesof the pH and of the mental conditions
wereplotted having the common time as abscissa.A striking correlation
betweenthe two curves was found in the first investigationsin manic patients.with the pH of the urine at higher values, the patient was calm,
whilethe more acid urine correspondedto periodsof intensiveagitation.
Figure33 showsan example.
This correlationalsocould be demonstrated
by administering
acidifying
andalkalizingsubstances
to manic patients.Acidificationthrough the administrationof phosphoric acid was followed by a manifest increasein
agitationwhile administrationof an alkalizingagent-sodium bicarbonate
76
REsEARcHrN pHysropATHoLocy
-was followed by a pcriod of calm. An oppositebut less evidentcorrelation was seen for severaldepressedpatients.we must mention that the
usual difficulty of judging accuratelythe degreeof depressionfrom one
hour to another can explain this lessercorrelation.Manic manifestations
were much more readilv evaluated.
o
(o
q)
o_
Io
(u
c 6o
L
Hours
Fro. 33. Tbe variations of the curve of the ogitotion of a lO-year-old boy are oppositc to that of the urinary pH, indicating an acid pattern.
DUALTSM
77
Dyspnea
Certain characteristicsof dyspnea suggestedthat this symptom might be
exploredby the srme meansemployedin studying pain associatedwith pathological tissue changes.A regular pattern is observed in certain types of
dyspnearelated to the time of day. Cardiac asthma or paroxysmal cardiac
dyspneaoccur mainly at night after patients have gone to sleep. However,
this is not due to the recumbentposition since patients may assumethis
position during the day without any ill effect. Harrison's "evening dyspnea"
(20) is characteristicallyabsent in the morning, but develops slowly during the course of the day, reaching a maximum in the evening.
Just as for pain, the degreeof intensity of dyspneawas studied in relation to hourly acid-basebalancechangesas indicated by changesin the pH
of the urine. Patientshaving dyspneaof prolongedduration as a result of
variouspathologicalconditions,with no treatmentof any kind for at least
six hours before or during the period of observation,were the subjectsof
this investigation.The degreeof intensityof the dyspneawas estimatedby
trainedobserverswho were in constantaftendance.Dyspneawas recorded
by the observersas absent,slight, moderate,severe,or very severe,estimationsbeing basedupon rate depth and evidentdifficulty in respiration.
At the time of these observations,hourly urine specimenswere obtained from patients with as little disturbanceas possible. No patient
showedevidenceof renal disease.The pH of the urine specimenwas determined potentiometrically.The curves showing hourly fluctuations in
the intensity of the dyspneaand the changesin the pH of the hourly urine
specimenswere then plotted and compared.Acidifying and alkalizingsubstanceswere administeredto patients during the course of some tests in
order to observe the influence of induced changesin the acid-basebalance
uponthe degreeof dyspnea.Phosphoricacid and sodium bicarbonatewere
usedfor this purpose.
Fourteen patients with different pathologicalconditions were studied.
In ten cases, a distinct correlation betweenthe intensity of the symptom
and acid-base variations was found.
Four patients had pulmonary edema associatedwith the symptom of
dyspnea.One patient had edemadue to congestiveheart failure; another
had pulmonary edema and lung metastases
from a carcinomaof the pancreas.Two other patients with cancer of the breast metastaticto the bone
andskin also had pulmonaryedema.In one of these,the pulmonaryedema
appearedto be a result of the accidentalintroduction of a fatty acid in oil
preparationinto the blood stream following an intramuscularinjection. In
78
n E S E A R C Hr N p H y s r o p A T H o L o c y
DYSPNEA
Seyere
Moderote
Stight
None
pH
Urine
6.5
6.O
5.5
5.O
4
Hour
Ftc. 34. The parallel variations between the curves of the intensity of dyspnea and
o f t h e v a l u e o f t h e u r i n a r y p H i n a c a s e o f p u l m o n a r y c o n g e s t i o nf o l l o w i n g a n
a c c i d e n t a li n t r a v e n o u si n j e c t i o n o f a f a t t y a c i d p r e p a r a t i o n ,i n d i c a t e s a n a l k a l i n e
patlern.
istrationof phosphoricacid, while sodiumbicarbonateincreasedthe dyspnea. By analogywith pain, we called this correlationan alkalinepattern.
(Fie.34)
Six casesshowedan oppositetype of correlationbetwecnthe intensity
of dyspneaand the acid-basechangesin the pH of the urine, All of these
DUALISM
79
Severe
Moderote
Slighl
None
pH
Urine
6.5
6.O
5.5
5.O
3
Hour
F r a . 3 5 , T h c o p p o s i t ec o n c o m i t a n tvariations betwcen the intensity of dyspnea and
urinary pH in a case of mcdiastinal metastasesof a hypernephroma indicatc an acid
paltefn,
80
REsEARcH tN pHysropATHoLoGy
oYSPNEA
NoHCOs
Seyerc
Moderotc
Slight
None
7.5
70
6.5
6.O
5.5
f
Hour
Flc. 36. The administration of sodium bicarbonatc increases the intensity of the
dyspneain a casc of pulmonary metastases
of cancer of the gall-bladder,indicatin3 an
alkaline pattcrn.
DUALtSM
8l
82
R E s E A R c Ht N p H y s r o p A T H o L o c y
DU^LtsM
g3
Flc" 37, The curve of oral temperutureof a casewith gcneralizedmctastaticmelanoma is pcrsistentlyabovc the 98.6'F (37'C) line, which corrcspondsto the averagevaluc of normal individuals.
Fre. 38. The curve of the oral tempcratureof a paticnt wilh gcneralizcdmctastases
of the breastshowsvaluesconstantlybelnx'lhe 9t.6"F averase
of adcnocarcinetma
line.
DUALTsM
r{i r1,..
R.
6-lo
O$r
IT
* 1 "
rr .,r,a rrrk
I
/3
/2
/ 8 5
t,g
/{
/6
/7
I
rl
!06
lt
L1r
rt
; ;
: "J
r'i
ri
tlJt
I st'
101"
t:
,
j ror' .",|
t
.J
rma
nt'
$'
it
t \
-,I
'./
I
\
t
\
\ia
Ftc. 39. Curve of the oral temperature of a patient witb bronchogenic cancer with
abdominal metastassshows values constantly helnw g8.6"F (37'C) avcrage line.
independentof type or site of origin of tumors. we will discussthe significaoce of tempefature patterns later.
.Syrtenrc Analyses:
Changesin urine or blood valueswere followed in hundredsof subjects
for long periods,even years. It appearedessentialto plot the data graphically. An average value, obtained from a significant number of normal
subjects,was riepresentedin graphs as a referenceline. In normal subjects,
as a general rule, relatively wide oscillations omur around the average
value line. By contrast, in diseasestates,there is a fixation of the curve on
one or the other side of the averagevalue, the curvesexhibiting only slight,
or even no, variations.Two opposingpatternsare thus evident in disease
sialesfor each type of analysisof blood and urine. The dualism indicates
once again the existencein systemicmetabolismof two kinds of abnormal
changeswith antagonistic characteristics.It is important to note that the
advancedcancerouscondition is characterizedby a marked dualistic pattcrn at the systemic level.
86
REsEARcHrN pHysropATHoLocy
Blood
In blood analyses,the concentrations
of potassiumand calcium, the
presenceof C reactiveproteins,the number of leucocytesand of circulating eosinophiles,(Nole 7) and the red cell sedimentationrate, have been
studied,usingclassicalmethods.They all show the samedualism.Figs. 40
to 43 show someof thesecurveswith the averagevaluesas referencelines.
with the intention of obtaining information concerning the amount of
potassiumpresent in cells, we investigatedthe content of this cation of
the red cells.(Note 8)
Urine
In urine studies, measurementswere made for pH, specific gravity,
surfacetension,oxido-reduction
potential,excretionof sodium,potassium,
calcium, chlorides, phosphates,sulfates, sulfhydryl, indoxyl, grucuronic
acid, peroxides, etc. Most studies were carried out by routine test techniques.For some analyses,however,conventionaltechniqueswere found
to be inadequateand new testsdevised.
For the urinary excretionof sulfhydryl,a new techniquewas devisedby
M. Bier and P. Teitelbaum in our laboratories.Using the warburg
micromanometer,
thc nitrogenliberatedfrom sodium azide in a buffered
solution in the presenceof free iodine was found to be directly related to
the amountof sulfhydrylpresent.The amountof nitrogenfreed at a determined momcnt-13 minutes-was most indicative.(Note 9)
For the information which we neededconcerningthe amount of calcium in urine a very simplemethodwas devised.(Note I0)
For measurementof urinary surface tension, we devised a new
technique.We used a capillary so calibratedas to give the surfacetension
in dyne/cm for a fluid with a specificgravity of 1,015. In this method,
severalarrestsor slowdownsof the descendingcolumn are noted and make
it possibleto obtain informationabout an extremelyimportant factor which
is usuallynot consideredin the measurement
of surfacetensionwith other
methods.It is known that urine is formed of differentconstituents.
some
of them with the tendencyto move toward the surfacewhile others tend
to move toward the bulk of the fluid. Changesin the distributionof these
constituentstake place. They induce changesin the value of the surface
tensionof the urine which occur even during the time measurements
are
taken. The descendingcolumn in a capillary will indicatethesechanges
which are important for precisemeasurement
of surfacetensionof urine.
(Note I I )
D U A L I S M
87
lz
!'
a1 a
Ftc. 40. Curve of the value of the K + in blood serunt in a case with periarteritis
nodosa. The values remain above 4.5 mEq, which represent the average value obtained from seriesof normals.
H
Y
5
c
o
a/,
F I c . 4 1 . C u r v e o f t h e v a l u e so f K + i n t h e b l o o d s e r u m o f a s u b j e c tw i t h c a r c i n o m a
of thc breast, liver metastasesand jaundice. The values remain almost constantly below the 4.5 mEq line.
,:{#:i;;!lil:tt;iiil:tiii'i1l,l1$.'*ii
{llii*ri'+
88
R E S E A R C H
I N
P H Y S I O P A T H O L O C Y
U'
O.J
t)
o
IJ
f
Days
F I o . 4 2 . T h e c u r v e o f t h c n u m b e r o f b l o o d l e u c o c y t e so f a c a s e o f b r e a s t a < J e n o carcinoma shows constantly values abovc 7.0O0,considercdas the average value for
normals.
9ao
C
7
(e
3n
(l,
(J
f,
(J
5
o,
-J
0ays
Ftc. 43. The curve of thc number of blood leucocytesof a casc of brcast carcinoma
showspcrsistentlyvalues below the averageline of 2.000.
DUALTSM
89
90
xESE^RcH tN pHysropATHoLocy
Sagiibr,
| l ? o ? 2
?r
0" 2 1
.o 15
9
AI
ERS
Flc. 44. Curves of. diflerent anall'sesof daily urine samples of a normol 4O-yearo l d m a l e . F o r e a c h k i n d o f a n a l y s i s t, h e c o r r e s p o n d i n ga v e r a g cv a l u e w a s c a l c u t a t e d
from the measurementsobtained in more than one hundrcd normal subjects. The
curves are seen to pass from one to the other side of the lines correspondingto thc
r e s p e c t i v ea v e r a g ev a l u e s .A c e r t a i n c o r r e s p o n d e n c ies s e e n f o r t h e c h a n g e sw h i c h
take place at the same time in these curves. Parallel variations are seen between spccific gravity and chloride index, while opposite to thosc for the pH and surface
tension,
9l
D U A L I S M
at;-./
'r_-.^..,"
-'.
.-_..
"-.'
&n
9n
I
(b)
t,
oon
pH
50
PH
6
I
6
Frc. 45. Curves of the valuesof the urinary pff followed during 50 days in five
s u b j c c t s .T h e c u r v e ( a ) c o r r e s p o n d st o a n o r m a l i n d i v i d u a l w h i l e t h e o t h e r f o u r
( b , c , d , e ) t o s u b j e c t sw i t b d i f f e r e n t c a n c e r o u sl e s i o n s .F o r t h e n o r m a l c a s e , t h e
values pass above and below the 6.2 value which is consideredas the average computed values obtained from normals, For the abnormal cases,the curves which show
l i t t l e v a r i a t i o n s a r e f i x e d a t o n e o r t h e o t h e r s i d e o f t h e a v e r a g el i n e , F o r c a s e ( b ) ,
an adenocarcinoma of the ovary' and case (c), a bronchogenic cancer, the curves
are fixed below the averageline. For case (d), with a bronchogeniccancer. and case
( e ) . a c a n c e r o f t h e b r e a s t w i t h g e n e r a l i z e dm e t a s t a s e st ,h e c u r v e s a r e a b o v e t h i s
averageline.
*s.8"3
.5-
E s $g s
: g. e8
E
! E8.r
( ! e
7 o X- q
o o
E 5 : -
[ i i
:u$iE
efldrr
6 ;
l E g
9;r
.!rsFE
i:l*:
: 8 " i
I ' I iT
n E !$e
-';3:
l! :$ .lg3e. t i
:F:;
;;[;E
D U A L I S M
/ e 3
July
June
?5
6ts
o
a 5
Po?ient-Mr: Spe
Ftc. 47. Curve of the values of the urinary specific gravity in a case of cancer
of thc breast, with the values fixed above 1016 which is considered as an avcrage
value for the specificgravity as computed from a seriesof measurementsin normals.
Moy
ro
r5
20
25
25
:15
a
\,/
---'---''
Potienf-Mrs FS.
F r c . 4 8 . T h e v a l u e so f t h e u r i n a r y s p e c i f i cg r a v i t y i n a c a s eo f c a n c e r o f t h e b r e a s t .
are the whole time below thc averatc value of 1016.
94
RESEARCH IN
PHYSIOPATHOLOCY
Ftc. 49. The urinary surloce tension remains constantly fixed abot,e the average
value of 68 for more than a year in a case of cancer of the urethra.
2oo
t7t
x
tu
'c,
c
L
E
t8,
/15
E
f
(-4,
loo
.75
.sat
0ays
F t c . 5 0 . T h e u r i n a r y s u l f h y d r y l i n d e x r e m a i n s b e l o w t h e a v e r a g ev a l u e o f
c a s co f c a n c e ro f t h e b r e a s t ,w i t h b o n e m e t a s t a s e s .
DUALISM
95
It was especia.lly
in advancedstagesthat definitegroupingsof patterns
with opposite characteristicscould be recognized.They correspondedto
two fundamentaloffbalanceswhich we have called "Type A" and ,.Type
D." 1"4" for anoxybiosis,"D" for dysoxybiosiswhich representthe principal manifestationsof oxygenmetabolismin a phaseof theseoffbalances.
)
The correspondencebetwecn the different abnormal patterns defining
the offbalanceA or D, is seenin Figures52 to 60. The abnormalpattern of
q,
!
s
L
-t)
E
t
L/'
Frc. 5 1 . T h e u r i n a r y s u l f h y d r y l i n d e x r e m a i n sa b o v e t h e a v e r a g ev a l u e o f r . 5 i n a
caso f h y p e r n e p h r o m aw i t h l u n g m e t a s t a s e s .
e 6 /
RESEARCH IN
t94l
Moy
30
PHYSIOPATHOLOGY
Ju n e
ro
r5
20
Io25
(9
l5
o.
a
rooo
7.4
o.
62
54
4.6
Polrenl- Mrs Von
Frc. 52. A correspondenceis seen between the patterns of. urinary specific grat,ity
and that of the urinary pH in a cancer of the colon correspondingto tbe type A offbalance.The changesstill present in the curves are opposite.
ZC
62
4 6
cl
I tiaar
2.5
F r c . 5 3 . I n a c a s e o f s a r c o m ao f t h e l e g w i t h l u n g m e t a s t a s etsh c c u r v e o f t h e a r i nury pH is fixed above the averageline (6.2) while the curve of chloride index Wl o w t h e a v e r a g el i n e o f 2 . 5 . T h i s r e l a t i o n s h i pindicates a type A of the offbalance.
D U A L I S M
Fab.n,t
5
r
/ e 7
-22
lo 't .2
18r
is: {-- 1
4CJ
323
Oro
t = ^
;..f-eo.l
F t c . - 5 4 .T h e a n a l y s i so f a t e r m i n a l c a s eo f c a n c e ro f t h e b r e a s t ,s h o w i n g t h e
fundanrcntaloflbolance, low specific gravity, high pH, low chloride index and high surface tension, correspondingthus to the offbalance A.
,n.-'n
i..
\_/\_,\
60
t'/
r/\r
Pgt,anl Arh
Ftc. 55. A terminal case of cancer of the breast shows all the analyses fixed in
opposite position to the case of Fig. 54, i.e., high specific gravity, low pH, high
chloride index and low surface tension, corresponding to an offbalance of the type D.
RESE^RcH IN
Leucocyt
es
o
. eg n
'8'
o
PHYSIOPATHOLOCY
(t,
E
ld
, *lllitRo',.id!t!s
(t
I
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)
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ly
q
Ftc. 56. offbalance type D, shows high urine specific gravity, tow pH, low surface
tension and low blood leucocyte number.
66
64
-o Q'
6.o
'A
:2t
-
a{,
.fe
/3'
io'
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56
Ftc. 57. The 3 curves, PH, sulfhydryl index and surface tension show an offbalance
type D.
DUALtSM
gg
isticphysiopathological
mechanism
studiedin complexconditions
in gcneral
andin cancerin particular.
r5
5?
so G. t 6
o
9H
7d
62
46
Cl
2 5
sl
74
68
@
Potranr Alr
F t c . 5 8 . T h e u r i n e a n a l y s e so f a p a t i e n tw i r h c a n c e ro f t h e o v a r y . s h o w i n g a d i s c o r d a n c e b e t w e e nt h e p a t t e r n sp r e s e n t .W h i l e a l l t h e a n a l y s e ss h o w p a t t e r n so f t h e
t y p e D , t h e c h l o r i d e i n d e x r e m a i n s t h r o u g h o u t t h e e n t i r e o b s c r v a t i o nc o n s t a n t l yo f
type A.
- U 3 c i <n
i;sHn
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e i I T*
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$ g ;E :
: 3 9 s E
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DUALISM
l0l
!2
d16
a
Ftc. 60. A condition can pass from one offbalanceto the opposite one. The passage
ol the analyses lrom the oflbalance lype D into the oflbalance trpe ,,1 is seen in a
caseof caocer of the lung. The analyseswhich first show high specific gravity, Iow
pH and high chloride index, pass subsequentlyto the opposite side, with low specific
gravity, high pH and low chloride index. This new oflbalance remains unchanged
for a long time as shown in the second part of the curve.
102
*ESEARcH IN
PHYsroPATHoLocy
Offbalance"A"
Offbalance"D"
Rapid cellularaging
Less difterentiatedcells, especially More differentiated cells, espeof connectivetissues
cially of connective tissues
lncreased amount of connective Decreasedamount of connective
tissue
tissue
Organic
Low
oxido.reduction poten
processes
Low chloride content
Local acidosis
Acid pattern symptoms
(Pain, dyspnea,itching,etc.)
Decreasedlymphatic tissue
High oxido-reduction potential
processes
High chloride content
Local alkalosis
Alkaline pattern symptorns
(Pain, dyspnea,itching, etc.)
Somnolence
Constipation
Polyuria
Exophtalmia
NasalpH below6.5
Slow absorbtionof skin wheal
Insomnia
Diarrhea
Oliguria
Enophtalmia
Nasal pH above6.5
Rapid absorbtionof skin wheal
Systemic Blood
Leucocytosis
Eosinophilia
High color index
Low potassiumcontent
Low R.C. sed.rate
No C reactiveprotein
Urine
Low specificgravity
High pH
High Cl, Na excretion
Low SH, Ca, phosphate,sulfate
excretion
High surfacetension
Death in coma
Blood
Leucopenia
Eosinopenia
Low color index
High potassiumcontent
High R.C. sed.rate
High C reactiveprotein
Urine
High specificgravity
Low pH
Low Cl, Na excretion
High SH, Ca, phosphate,sulfate
excretion
Low surfacetension
Presenceof oxidizing substances
Death whi.leconscious
DU^LrsM /
103
CHAPTER
TH E CONSTITUENTS
E-lr
.lL lltsnencHrc oRcANrzArroNand dualism have opened the way for
a
study of the body constituentsin an attempt to systematizethem and their
functioningin accordancewith thesetwo concepts.In this research,we
consideredbesidesthe constituents
separatedin groupsas lipids, proteins,
carbohydrates
and electrolytesalso the elementsas a sourceof interesting
information.
With most of the manifestations
integratedin dualistic patterns,rvc
plannedto test the differentconstituents
by noting their influenceon these
patterns.It was to be expectedthat some might have selectiveactivity at
certain levelsof organizationand the manifestationsrelatedto theselevels.
This selectivitydid becomeevidcntbut it also turned out that any agent,
in sufficientamount, cxertcd an effect at any level. The problem was to
selectthe manifestationwhich would respondmost readily to the greatest
number of agents.This would make comparisonsbetweenagentseasier
and serveas a practicalcriterionfor the start of classification
on this dual
basis.
The measurementof the influenceexertedby various agentsupon the
secondday wound crust pH (s.d.c.pH) proved to be particularlyrewarding and was employedin the first part of the investigation.
The s.d.c.pH
providedan indication not only of acidifyingand alka.lizingeffectsbut also
comparative values for these effects. Later, the influence exerted upon
many other manifestations
was studiedfor corroboration.Details of the
s.d.c.pH techniqueand thc resultsobtainedare in Note 1.
In studyingthe elements,we chose,first, simplecombinationsin which
they appear.Each anion was investigated
by studyingit as it occurredin
thc respective acid and in compounds in combinations with diffcrent
cations;each cation was studiedin its combinationswith differentanions.
104
THE CONSTITUENTS
105
106 /
xESEARcHrN pHysropATHoLocy
rHE
CoNSTTTUENTS /
107
bers (II, IV, VI); amongall the B series,in thosewith odd numbers(I B,
III B, V B, VII B); and, amongthe C series,in thosewith alternatenumbers which can be consideredto correspondto even numbers. All these
series-with partial fulfillment for the sum of their shells-have a D inducing or anti A, character, and thus a homotropic tendency.
Periods
Coming back to the influenceexertedby the elementson test manifestations, we found that elementsof the same seriesshow some similar
propertieswhen acting at a certain level of organization,but some differencesappearwhen they are acting at difrerentlevels.Someof thesedifferences are important. Magnesium,calcium and strontium act similarly on
the s.d.c. pH, and againstconvulsionsas well. However, magnesiumhas
been found to induce somnolenceor even deep sleepin test animals,while
calcium immediately wakes them from magnesium-inducedsomnolence.
This type of antagonisticaction among membersof the sameseriessometimes apparsespeciallypronouncedbetweentwo consecutivemembersin
the series-for example,betweensodiumand potassium,magnesiumand
calcium,oxygenand sulfur, and sulfur and selenium.
Study of this "antagonism" has permitted us to recognizea specific
characteristic.When two elementsof the same seriesact upon the same
entity, one may substitutefor the other.Sodiummay replacepotassiumin
cells.Magnesiumand calcium.oxygenand sulfur,and sulfur and selenium
can replaceeachother in this kind of reciprocalactivity.There is no truly
antagonisticaction betweenthem. This explainsthc fact that two elements
of the same series,if in sufficientamount can have similar activity at a
given level-that of the tissular,for instance,where the changesof the
s.d.c. pH take place.
Further analysisof the activityof membersof the sameserieshas revealedanotherimportantcharacteristic
which has permittcdfurther classification. Differencesin activity of membersof the same seriescould be
related to the organizationalcompartmentsinvolved.This becameclear
when activity of members of the I A series was analyzed according to
whether these elementsform constituentsof the metazoic,nuclear or
subcellularcompartments.
Sodiumis the predominantcation of the metazoiccompartment,which consistsof the interstitialfluids,lymph and blood.
Potassiumis the principalcationof the cellularcompartment.Ammonium,
which corresponds in most of its propertiesto rubidium, representsthe
cation at the nuclearlevel.It could be seenthat the developmentof hier-
108
nEsEARcH rN PHYSToPATHoLocY
THE CoNSTtruENrs
lo9
plv.
The generalrule, which appearsto governthe variationsin distribution
of an elementwithin the organism,makesit important to know the proper
level of an element. Some exampleswill illustratewhat we mean, An increaseof copper is seen in the blood serum of cancer patients,althougb
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THE CoNSTrruENTs
lll
ll2
RESEARcH rN PHYsroPATHoLooy
CHAPTER
1r
pATHwAys,one theoreticaland the other experimental,
lLrvo DTFFERENT
have led us to considerlipids as possiblythe most important constituents
involved in thc dualisticpatternsof physiopathological
manifestations.The
study of all the constituentsof the organism--electrolytes,
proteins,carbohydrates and lipids-has shown that for each of them, a rough division
into two classeswith antagonisticreactivitycan be made accordingto the
positive or negativeelectrostaticcharacterof their polar groups-nucleophilic or electrophilicfor some,anionic or cationicfor others.However,
these fundamentaldifferenceswhich can explain thcir interventionin
processesin which dualism is apparent,do not representthe reason for
their rolc in the inductionof patterns.
The reactionsin which someof theseconstituents
take part are carried
out as rapid changeswhile others are complet.,ed
only slowly. It is these
slow rcactions,once accomplished,
which tend to be stablefor long periods
of time. Sinccsuchstabilityis characteristic
of clinicaland analyticalmanifestationswhich have dual patterns,it appearedlogical to considerthe
with slow reactivitywhich are relatedto thesemanifestations.
constituents
Becauseof their hydrosolubility,
and the rapidityof the reactionsin which
part,
proteins
take
most
they
electrolytes,
and evencarbohydrates
probably
play a lesserrole in theselong lastingprocesscs.
The lipids, on the contrary,seemto be especiallysuitedfor this role.
Many of the reactionsin which the lipids participateare slow. As we will
seelater,this is primarily becauseof their insolubilityin water.They form
in the organisma group "apart" from all the water solubleconstituents,
a fact which permits them to function through proper reactions largely
without continuous interferencefrom thc other constituents.For these
reasons,the lipids appearedto be the most likely of all constituentsto be
lt3
114 /
R E S E A R c HI N P H Y S t o P A T H o L o c Y
OF LIPIDS
LTPTDSAND
LTPOTDS
ll5
Hydro
id
Borderl
ine
Lipoid
FN\\\
FI\\\\\\\\\
eot.rGroup
Group
Sl llon-Polar
a certain apprehensionfelt by many workers about incorporating indistinctly, in the same group of agents,substanceswith vastly different chemical constitutions,which until now have not been associatedwith lipids.
The fact that, beyond physicochemicalconstitution, biological properties
characterizingthe lipids are common to the entire group of lipoids, will
in time, we hope, help to reduce the importance of this separation betweenlipoids and lipids.
The structure of the lipoids-with a large variety of polar and nonpolar
Sroupsbut always with the same characteristicenergeticrelationshipbetweenthem-has led to a logical systematizationof thesesubstances,using
thenature of the polar and nonpolar goups as criteria.
CLASSIFICATION OF LIPOIDS
l16
nESEARcH rN pHysropATHoLocy
4.
5.
6.
7.
8.
9.
10.
Lipeamines
Lipo-amides
Lipo-alcohols
Lipo-aldehydes
Lipo-ketones
Lipo-halogens
Lipo-metals
etc.
( -NH.)
-coNH,)
-oH)
-cHo)
(:CO)
(-Cl, etc.)
( -Na, etc.)
(-SH)
( -so"H)
(:S)
(-so)
(:SO,)
(:SO")
2. Lipo-nitrogenderivatives
a.
b.
c.
d.
e.
Lipo-amines
Lipo-amides
Lipo-nitriles
Lipe.isocyanides
Lipo-nitro derivatives
etc.
(-NH,)
(-CONH,)
( -CN)
(-NC)
( -NO,)
L t P t D s A N D L r P o t D S
l 1 7
accordingto theircarhttnbond.s.
B. Lipoidscla.ssified
l. Saturated
2. Unsaturated
a. Ethenic(mono-,di-,poly-)
b. Ethynic
Someaspectsof this classification
requirediscussion.
The genericterm lipoacidhas beenemployedto describesimplelipoids
having polar groupswith acid functions.While the principallipoacidsare
the fatry acids,other membershavc other acid polar groups,such as SO3,
SH, NOr, etc. The significanccof this groupingtogethcrof lipoids with
negativepolar groupshas becomeevidentespeciallyin studyingthe similaritiesin the biologicaleflectsof thcsesubstanccs.
In certain aspectsof
our research,this correlationhas pcrmittedus to substituteone category
of lipoids (lipo-thiols or lipo-aldehydes) for another (lipo-carboxylic
acids), therebyavoidingcertain undesirableeffectsof the latter group of
substances.
Lipoids havingpolar groupsenergetically
oppositeto thoseof the acids
have been grouped togcther.O[ thcse,the memberswith a polar group
with alkalinefunctionshave becn classifiedas lipobascs.The tcrm baseis
generallyappliedto ionizablecompoundswhich influcncethe pH of solutions and combine readily with acids by losing an OH and gaining a
proton. Another group is formedby the lipoidicalcohols.Recentevidence
indicatesthat, in miiny circumstances,
the differencesin the reactionsof
alcoholsand common basesare quantitativeratherthan qualitative.Quite
often the reactionof an alcohol with an acid is analogousto the reaction
befweenan acid and sodium hydroxide,thc H' of the acid combiningwith
the OH- of the alcohol.The differcnces
betweenreactionsare considered
to be mattersof time-rate.Whereasthe reactionof the baseis almost instantaneous,
that of alcohol is a slow reactionand is less complete.This
behavior of alcoholic substancesis particularly clear when the hydroxyl
group of an alcohol is replacedby a halogento preparethe alkyl halides.
For instancc,accordingto Karrer (24): "This can be done by the action
halogcnacidson the alcohol:
of the concentrated
CH3OH + HCI -----> CHrCI + H2O
The reaction correspondssuperficiallyto the formation of a salt from an
acid and a base:
NaOH + HCI ------>NaCl + H:rO
There is, however,a differencebetweenthe two processes.
Basesand acids
are largelydissociated;when thcy come together,hydrogenions and hy-
ll8
,/
R E S E A R c HI N P H Y S t o P A T H o L o c Y
droxyl ions combine almost at once to give the very little electrostatically
dissociatedwater, so that the reactionwhich really occurs is:
Nal * OH- + H+ + Cl- -----> Na+ * Cl- + HrO
Ionic reactionsalwaysoccur instantaneously.
Reactionbetweenalcohol
and hydrogen halides is governed by other laws. Alcohol is only very
slightly ionized. For the removal of the hydroxyl group a certain time is
required.The reactionbetweenalcoholand acid with eliminationof water,
known as esterification.
is thereforea time reaction."No essentialdifference exists between the reaction of lipo-alcoholsor lipo-amines,for instance,with organicacids.
Theseconsiderationswould have been sufficientto allow lipobasesand
lipo-alcoholsto be grouped together.There are othcr considerationsas
well. Their common biologicalactivity and mutual interchangeability
also
justify
appcar to
grouping them together.Furthermore,the recognitionof
the existence
of a generalmutual antagonismbetwecnlipoacidson the one
hand and lipobasesand lipo--alcohols
on the other hand, chemically,physically and biologically,has proven of considerablevalue in explaininga
variety of experimentallyobservedfacts in many aspectsof our research.
Following this through, we have found it advantageousto define the
two groupsof lipoids by a more generalcharacter,the electricalaspectof
the polar part, negativcfor the lipoacidsand lipo-aldchydes
and positive
for the group of lipo-alcoholsand lipobases.The terms,"positive and negative lipoids,"servealso to emphasizethe natureof their antagonism.
The structureof thc nonpolargroup as it confersphysical,chemical
and biological propertieson the lipoids permits further subdivisions.Lipoids may be classifiedon the basisof thc a.liphatic,alicyclic.aromaticor
heterocycliccharactcrof the nonpolar group. While the negativelipids
are principally formed by fatty acids,the positive are made up principally
of sterols.The prcsenceor absenceof doublebondsdefiningsaturatedand
unsaturated
carbon chainshas becn one subjectof our study and considerablebiologicalimportancehas bcen found to be relatedto this character
as well as to the positionalrclationshipof the double bond to the polar
group and the polarity inducedby the doublebond.
The study of lipoids has shownthat, bcsidespropcrticscontributedby
the elementsand groups which compose thcm, they have additional
physico-chemical
and cven biologicalpropertieswhich are characteristic.
We have termedthcse"lipoidic propcrties"to indicatethat they are consideredto resultdirectlyfrom the particularconstitutionof the lipoids.
LIPIDS AND
LIPOIDS
II9
120 /
R E S E A R , C HI N
PHYSIOPATHOLOGY
TrsLe VI
CursstntcerroNoF Cueulc,rl Coupouxos
Composition
Predominance
Name
Polargroupsonly
Polar-nonpolar
groups
Nonpolar groups
only
Example
Water
Polar group
predominant
Hydroides
Glycerin
No predominance Borderline
substances
n-Propyl alcohol
Nonpolargroup
predominant
Oleic acid
n-Butyl alcohol
Lipoides
Paraffin
The "lipoids," in which the nonpolar groups predominate,are more soluble in neutral solventsthan in water.
As we have mentionedabove, from a practical point of view, a substancecould be judged to be a hydroid, borderlinesubstance,or lipoid by
consideringthe differencesin its solubility in water and in a nonpoliu solvent, such as petroleumether, which correspondsto a mixture of the first
aliphatic saturatedhydrocarbonsliquid at normal temperatureand pressure. ,{ polar-nonpolar substancemore soluble in water than in neutral
solvent is considered a hydroid; one equally soluble in both solvents is
classifiedas a borderline substance;w,hilea substancemore soluble in the
neutral solvent than in water is a lipoid.
Difterent polar groups such as COOH, OH, NH2, CO, SOz, SH, etc.,
enter into the constitutionof various lipoids. They differ considerablyin
their electrostaticforces. As a result, the forces of the nonpolar goups
required for predominance,if a lipoid is to be formed, also will difter. A
different nonpolar group thus is necessaryfor each different polar group.
For aliphatic molecules,it is principally the length of the chain which
determinescohesionforces and a differentnumber of carbons in the nonpolar group appearsto be necessary,dependingupon the polar group, in
order to form a lipoid. The study of homologousseries from this point of
view is interesting.
Sincethe value of the electrostaticforcesvariesgreatly from one polar
group to another, the first members of the various homologous series.
which are also lipoids, will differ from seriesto series,dependingupon
L T P T D S^ N D L t P o t D S
l2l
the nature of the polar group. The lengh of the carbon chain of the nonpolar group will thus indicate in what member of a series the lipoidic
characterappears.By comparingmathematicallythe value of the electrostatic forces of each polar group and the cohesionforces of the nonpolar
group in the respectiveseries,it is possibleto determinewhich member of
each homologousseriesof substanceswill first show the properties of the
lipoids. This a.lsocan be determinedexperimentally,as seen above, using
the solubility characteristicsof the lipoids. For the different membersof
the series,degreesof solubility in a polar solvent such as water, and in a
nonpolar solvent such as petroleum ether, were determined.The first member of an homologous series to be considereda lipoid was the one found
to be more soluble in the nonpolar than in the polar solvent.All members
with a large number of carbon atoms show lipoidic properties;those with
fewer carbon atoms lack those properties.
Thus, lipoidic properties first become manifest, among the carboxylic
acid series,in valeric acid,i.e., the five-carbonmember.The shortercarbon
chain membersare soluble to an equal or greaterdegreein water than in
petroleum ether, while those having a carbon chain longer than four show
a higher degreeof solubilityin the nonpolarsolventsthan in water. (Tnnr-E
vlr)
TesLe VII
Sor,usrLlrresor CrnsoxyLtc Aclo HovolocuEs
ility in
Substance'
Common Name
Methanoic acid
Ethanoic acid
Propanoic acid
Butanoic acid
Pentanoic acid
Hexanoic acid
Heptanoic acid
Octanoic acid
Formic acid
Acetic acid
Propanoic acid
Butyric acid
Valeric acid
Caproic acid
Enanthic acid
Caprylic acid
Polar
Solvent
( W a t e ra t 2 0 " )
I
2
3
4
5
6
7
8
Nonpolar
Solvent
( Petroleum
Ether)
insol.
cO
3 . 7( a t 1 6 " )
0.4
o.24
0 . 2 5( a t 1 0 0 " )
' Namesapproved
by International
Unionof Chemistry.
The sameis true for the trJkylalcohols.n-Propyl alcohol and the members below it are either miscible with both water and petroleum ether or
more soluble in water, indicatingthat the nonpolar forcesdo not predomi-
?.:.]l,:'j
\jii;
122
RESEARcH rN pHysropATHoLocy
Trnle VIII
Sor-uslI-rrresoF THE Ar-xyr- Alconor-s
7o of solubilitvin
Substance'
Conrmon Name
Methanol
Ethanol
I -Propanol
2-Propanol
l-Butanol
2-Butanol
2-Methyl, 2-propanol
2-Methyl, I -propanol
l-Pentanol
2-Pentanol
3-Pentanol
2-Methyl, 2-butanol
2-Methyl, l-butanol
3-Methyl, 2-butanol
l-Hexanol
2-Hexanol
3-Hexanol
l-Heptanol
l-Octanol
Methyl alcohol
Ethyl alcohol
Propyl alcohol
Isopropyl alcohol
n-Butyl alcohol
sec.-Butylalcohol
tert.-Butyl alcohol
Isobutyl alcohol
n-Amyl alcohol
sec.act. Amyl alcohol
Diethyl carbinol
tert.-Amyl alcohol
n-act. Amyl alcohol
Isoamyl sec. alcohol
n-Hexyl alcohol
sec.-Hexyl alcohol
Ethyl propyl alcohol
n-Heptyl alcohol
n-Octyl alcohol
No. of
Carbon
Atoms
Polar
Solvent
(Water
at 20")
Nonpolar
Solvent
(Petroleum
Ether)
I
2
4
4
4
5
5
5
5
5
5
6
6
6
8
7.9
t2.5
@
9.5
2.7
5.3
insol.
t2.5
insol.
sl. sol.
very sl. sol.
very sl. sol.
0.9
insol.
insol.
co
oO
co
oO
@
co
&
*
t
*
cc
r
co
LrPrDsAND LrPorDs
123
Conrnron Name
N{ethanethiol
Propanal
Propylcarbylamine
l-Butanol
2-Butanone
Butanamide
2-Methyl, 2-butanol
Pentanoicacid (n)
H e x y l a m i n e( n )
l, I Octandiol
Methyl mercaptan
Propionaldehyde
Propyl isocyanide
n-Butyl alcohol
Butyl ketone
Butylanride
tert.-Amyl alcohol
Valeric acid
Hexylamine
l. 8 Octandiol
Polar
Group
-SH
-CHO
-NC
-oH
:CO
-CONH.
-oH
-COOH
-NH'
-oH
i
]
No. of
Carbon Atoms
l
J
J
A
,
A
a
5
5
6
8
t N a m e sa p p r o v e db y I n t e r n a t i o n a l
Union of Chcmistrv.
124 /
REsEARcHrN pHystopATHoLocy
F t c . 6 2 . T h e p r e s e n c eo f t h e s a m e e l e c t r i c a lc h a r g e a t t h e s u r f a c eo f d r o p l e t so f a n
emulsion insures,througb the repellent forces, the stability of the emulsion.
Chemical Properties
Lipoids have two groups of chemicalpropertieswhich can be related to
their two principal parts, polar and nonpolar. The polar groups with their
electrostaticforces give to the lipoids one group of characteristicreactivities. A carboxylic lipoid will act like any other organic acid, while the
lipo-'alcoholswill act like other alcohols,a thiolipoid like a mercaptan,
and so on. A characteristicof chemical reactionsinduced by the polar
groupsis that, while they are occurringin a water medium, they are largely
limited to the site where the lipoid is localizeddue to the insolubility of
125
the entire molecule in water. Through this localization, the polar reactivity
of the lipoids becomeslargely a "surface reactivity." It is interestingthat
even minute amountsof lipids are able, through this localizationat separating surfaces,to induce important changes.
A second group of reactionstake place at the nonpolar group and
especiallyat the different formations presentin it, such as double bonds,
cycles, etc. The hydrophobic and lipophilic character of the nonpolar
groups confers a specialcharacteron thesereactions.Most of them take
place in nonionic nonpolar media. Many occur at the semipolar double
bonds with nucleophilicor electrophiliccarbonswhich appear to be especially suitablefor this reactivity. This would explain the fact thar nondissociatedmoleculesmay take part in thesereactions.Most of thesereactions
are relatively slow. This double reactivity,ionic through the polar group
and rather nonionic through the nonpolar group, makesthe study of these
Iipoids one of great interestand it will be discussedbelow in more detail.
Biological Properties
The biological properties of lipoids in general also can be related directly to their physiochemicalcharacteristicsand, thus, to their peculiar
constitution.
Lipidic System
The relative insolubility of the lipids in water and their solubility in
neutral solventshas permitted us to separatethesesubstancesas a group
from the other constituentsof organisms.For more than just didactic purposes,we considerlipids to constitutea separatesystemin the organism.
The part played by lipids in the organizationand the functioningof various entities supports this concept.For example,when a lipoid is introduced into the organism, it will be selectivelydissolved in, circulated
through, retainedby and metabolizedas part of the lipidic system.overtone's"Index of Repartition" of anestheticsin the organism can be seen
to be a direct corollary of the existenceof such a systemalthoughthe anestheticagent can be a lipoid or a nonpolar substance.
A great degreeof independenceof this system is morphologically evident as in adipous cells, when fats circulateas chylomicronsor when
they form oriented layers. We have seen above how the orientation of
lipoids at the surface of water results from the relationshipbetweenthe
solubilitiesof the two constituentgroups,polar and nonpolar. Along with
126 /
s,EsEARcH
rN pHysropATHoLocy
LtPTDS AND
LTPOTDS
127
-ll+llllll
ooooooooc'
Fattyacidorientedlayr
ratgr
(b)
o
/z///,///,.
rater
oooooooo
Sterol orientedlayer
l.-
polar group
rrcn-polargroup
to this surface,the layer which they form exhibits no permeabilityproperties.It evenopposesany passage
throughit. It seemsthat fatty acidsand
sterolsmake separate"spots" in the cellularmembranesso that, through
their quantitativerelationships,
they confer differentdegreesof permeability to differentregionsof the mcmbrane.The changesin permeability
which resultfrom the antagonistic
interventionof the two groupsof lipids
seemto play an important role in norrnaland abnormal physiology.
With fatty acids inducing permeability,and sterols opposing it, the
fundamentalcharacter of their biological relationshipcan be recognized.
It would seemthat part of the function of sterolsis to opposethe activity
of fatty acids.Conceptually,sterolswould appear,in this specificactivity,
to be "anti-fatty" acid agents,with a capacityto control the activity of fatty
acids rather than be active by themselves.
Partly for this reasonas well as
for greater general understanding,it is necessaryfirst to investigatefatty
acid activity.
'EllElt;tlf
[Ln[it8.AIf
128 /
nESEARcHrN pHystop^THoLocy
FATTY ACIDS
Besides their constructive role in cstablishingboundary formations,
fatty acids appear to serve various othcr purposesin the organism.They
can be used as caloric metabolites,and they play an active functional role
in a biological change. while all fatty acids may exhibit these three
activities---<aloric,constructiveand functional-there are important individual difterences.with the carboxyl as common polar group, the differencesbetween the various fatty acids can be related to the nonpolar
groups.we will discussthis aspectof fatty acids, emphasizingonly what
can be consideredto be new contributionsto understandingthe biological
role of the substances.
Rancidity
The srudyof changeswhich take placein virro, on lipids and especially
on fatty acids after they have been separatedfrom the organisms,led us
to consider a possible parallelism between them and the changes which
take placein the organism.We tried thus to utilize especiallythe knowledge
furnishedby the study of the chemicaldeteriorationof natural fats generallyknown as rancidity(28), to betterunderstandand alsoto systematize
many of the processes
occurringin vivo.
Three typesof rancidityare describcd.In onc-hydrolytic rancidityfats are separatedin free fatty acids and glycerol (or mono- or di-glycerides), through thc intervcntion of lipolytic enzymes. These are often
producedby molds (Penicillium,Aspergillus,ctc.) or by microbesrich in
suchlipolyticenzymesor cven thc lipasepresentin the tissuesfrom which
the lipids are obtained.]-he characreristic
of rhis typc of rancidityis the
interventionof enzymesand the appearance
of frce fatty acidsas a result.
In a secondtyp. of rancidity,also occurringunder the interventionof
enzymes,an oxidativeprocessis involved.Thc characteristic
of this typc
of rancidityis that it affectsaJmost,if not cxclusively,saturatedfatty acids,
convertingthem into methyl-ketones
by a bcta oxidation process.This
"perfume rancidity" called so becauscof the odor of the methyl-ketones
with seven,nine or cleven carbonswhich rcsult-takcs place apparently
through the interventionof a peroxidaseprescntin certain molds (such
as penicillium glaucum) , one of its characteristicsis that it occurs
e s p c c i a l loyn s a t u r a t e fda t t y a c i d sw i t h a l o w n u m b c ro f c a r b o n s( 8 t o l 2 ) .
The third type of ranciditygroupstogethcrthe oxidativechangeswhich
take placeat the unsaturatednonpolargroup of the lipids. As they result
from the interventionof double bonds, the reactionsdiffer accordinsto
L T P T D SA N D L t P O t D s
Lzg
130
RESEARcH IN
PHYSIoPATHoLocY
have two uses: one, to reducethe melting point below body temperature
and thus permit easymobilization,and two, to induce changeswhich lead
to the breaking up of the long moleculeinto two shorter ones which can
be metabolizedthrough the Knoop oxidation.
All the data indicate that this fission would not take place at the double
bond but through a more complex process.A first change consists of
oxygen fixation at the carbon near the double bond. This leads to the appearanceof a hydroperoxidegroup. It is only in a subsequentstep that
the moleculebreaks at a place betweenthis carbon near the double bond
and the double bond itself, resulting in the appearan of short chains
which have an even number of carbonscapableof being directly metabolized through the beta oxidation.(Note 4)
The positionof the doublebond in the naturally occurringmonoethenic
fany acids, separatingalmost always a group of nine carbons toward the
carboxyl or the mcthyl end, (Note 5/ acquiresa special significancefor
the breakingdown of the moleculesfor caloric purposes.
The desaturationof the saturatedfatty acids,which would representa
first step toward allowing them to participate in metabolic caloric changes,
would usually takc place in the liver, apparentlythrough the same processesby which polyunsaturatedfatty acidsare partially saturated.(Note 6)
An interestingpart of the caloric metabolismof the saturatedand
monoethenicfatty acids, which will be shown below, is their combination
with glycerol to form triglycerides.
Constitutional Role
Although saturatedand monoethenoidfatty acids enter into the formation of boundary membranes,the di-, tri- and tetraenicmembersseem to
havea particularlyimportant role in the constructivefunction of fatty acids.
Some of them entcr directly into the formation of the membrane;some
forrn complex lipoids such as lecithinewith the glycerophosphoricradical
and nitrogen containing bases.As a rule, these last representa lipoidic
substratewhich would act as a neutral natural solvent present in membranes,and as such, intervenein the realizationof a diphasic medium at
the level of the boundary formation.This medium would largely insure
the orientationof the fatty acidsat the separationsurfaceand the formation of permeablelipidic layers,
FunctionalRole
The third role of fatty acids is as functional agentstaking part in certain reactions.This activity appearsto be strongly related to two factors:
LTPIDSAND LTPOTDS /
l3l
the presenceof an uncombinedcarboxyl $oup and the energeticintervention of the double bonds of the nonpolar part of the polyunsaturated
members.
Free fatty acids appear to be functionally active while the combined
ones usually are inactive.The activity is relatedonly partially to the direct
capacity of the carboxyl to realize new combinations.It resultsfrom the
induction exertedby the carboxyl upon the nonpolar group. The so-called
free fatty acids of the organism are probably bound in a labile form to
proteins, but this bond will not influencethe induction effect exerted upon
the nonpolar group. The intensive positive carbon of the carboxyl, together with the zig-zagdispositionof the fatty acid molecule,causesthe
inductive effect to charge the successive
carbons of the chain. They will
thus show alternativesigns.The even carbonsshow a negativecharacter,
while the odd ones are positive.The fact that oxygencombineswith positive carbonsexplainsnot only why, as in Knoop oxidation,this bond occurs
at C3, which is strongly positive,but also explainsthe so.calledalternate
oxidation (33) where the other following odd carbonsare binding oxygens.
Through the influenceexerted by the carboxyl, the double bond shows a
specialactivity which has been worth studying.
Double Bonds
There has been some tendencyto regardthe double bond as a weak,
easily broken point of the molecule.Actually, it emergesas an important
center of activity. With its capaciry to become a semipolar center, and
consequently,to bind or lose radicals,the double bond is an energetic
center in the molecule.Is importantcharacteristic
is the ability to effect
such changeswithout altcringthe chain of the moleculeitself. Since this
type of reactionis reversibleand can be repeatedfor the same molecule,
the double bond appearsto representa functional entity. Becausethe reaction principally involvesnonmetallicelements,the unsaturatcdfatty acid
takesan activepart in the metabolismin which theseelementsappear.
The study of rancidity has helpedus, by analogy,to systematizeoxidaas they take placein vivo. In addition to Knoop beta, several
tion processes
other g'pes of oxidation could be recognizedin which double bonds intervene more directly. The double bond, with its semipolarcharacter,influences nearby carbons, rendering them highly reactive. In one form of
oxidation, a molecularoxygen is bound to a nearby carbon to produce a
formation,as was shownto occur in vitro by Farmer. (31)
hydroperoxide
this oxygenfixationbecomesreversible.
When,under certaincircumstances,
the fatty acid will liberate the oxygen. It appearshighly probable that in
132 /
R E s E A R C Hr N p H y s r o p A T H o L o c y
L T P t D SA N D L t P o t D s
133
duration,could hardly be attributedto variationsin the quantityof the interveninglipids alone. More probably they would result from changesin
the nature of the interveninglipids themselves.
We have investigated
this
aspectof the fatty acids presentunder abnormal conditions.
As a guide for the direction to be followed in theseinvestigations,we
used the information furnishedby the study of rancidity. We believedrancidity would be ableto indicatebroadly the natureof the qualitativechanges
which the lipids may undergounder abnormalconditions.Conceptually,the
abnormal can be consideredto result from a loss of the capacity of the
organism to sufficiently control occurring processesand keep them in
the frame of the constantswhich characterizethe entity. Due to this lack of
effectivecontrol, the in vivo occurringchangesunder abnormal conditions
would closelyapproachthose which take place in vitro where such a control doesnot exist,Theseconsiderations
led us to searchfor changessimilar
to those seenin rancidity,or occurringin vivo in lipids, under abnormal
conditions.
As mentioned above, in rancidity a first group of changesconcerns
the polar group. Someof them result in the appearanceof free fatty acids,
others correspondto changesin the carboxylsthemselves,while still others
are representedby processes
of oxidationwhich occur in the chain near the
polar group. A secondgroup of rancidity changesconcernsthe nonpolar
group and especiallythe energeticcenterspresentin it, the double bonds.
The study of this last group of changesled us to consider,the changes
appearingin vitro under the direct influenceof heat and oxygen.As part of
thesechanges,we consideredof specialimportancethe conjugationof the
double bonds seen to occur in vitro as a step in the oxidation of polyunsaturatedfatty acids.This conjugationcorrespondsto a characteristicdisplacementin the moleculeof two or more of the double bonds present,so
as to result in parallel reciprocalpositions.
While in the simple bond two tetrahedriccarbons are bound through
their peaks,in the double bond they are bound by one edge, and in the
triple bond by a surface.In the conjugatedformation, the common edges
of two doublebonds,being separatedby one simple bond, are consequently
parallel.The planesin which the electronsof thesedouble bondsare moving
for eachdouble bond and which are perpendicularto that of the bond itself,
becomeparallel. Through the resultingreciprocalinduction their energetic
value is enhanced.
We have studied systematicallythe different qualitative abnormalities
concerning the fatty acids, guided mainly by the information obtained
throughthe study of rancidity.
.^
134 /
nESEARcHrN pHysropATHoLocy
LIPIDS AND
LIPOIDS
,/
I35
In abnormal metabolic changes,an important factor is the intervention of abnormal fatty acids in the metabolismof chloride ions, producing
an especiallystrongfixation of the chlorideion to the carbonsat the double
bonds. The conjugateddouble bonds in a fatty acid moleculeappearto be
especially suitable for this since an abnormal, irreversible fixation of
chlorides occurs in two steps. First, the halogen is fixed at the extreme
carbons of conjugatedformationswith a displacementof the double bond
in the intermediaryposition. In the secondphase,the fixation takes place
in the intermediarycarbons,too. (Nole 8D)
Functionally,fatty acids induce activationof oxygenas a norrnal process, but the appearanceof peroxidesor irreversiblefixation of ctrloride
ions is an abnormal event.
It is the abnormal fixation of chloridesby the conjugatedfatty acids
which leads to a more complex group of processcsinvolving sodium
chloride metabolism.with the chloride ion fixed. the sodium ion of sodium
chlorideremainsfree to enterinto othercombinations,
especially
with a carbonateion, producingstronglyalkalinecompounds.This processexplains
the appearanceof local alkalosisas a result of thc intcrvention of conjugated abnormalfatty acids,corresponding
to the chloride phaseof ',D."
The division of fatty acids into four groups-l ) saturatedand monounsaturated,2) di-, tri- and possiblyalso tetra-unsaturated,
3) tetra- and
higher polyunsaturated,and 4) conjugated---{orresponds
schematicallyto
the four principal roles---caloric,organizationa.l,functional and pathogenic
-which fatty acids play in the organism. These roles are seen to be
dictated both by the differentstructuresof the fatty acids and the different
substancesto which they are preferentiallybound. The fate of a fatty acid
in the organism seemsto be greatly influenced by its bond to other substances.As already noted, we have called these other substances"antifattv acids."
THE
ANTI.FATTY
ACIDS
135
nESEARcH rN PHYStoPATHoLocY
also found in reserye in adipous cells as triglycerides. The di-, tri- and
even tetraenic fatty acids usually enter the circulation as phospholipids,
that is, in direct combination with glyccrophosphoric ions. The polyunsaturated acids are bound to sterols when they enter the blood, circulate
and are stored. While the structures of the various fatty acids determine
their difterent roles in the organism, it is the anti-fatty-acid constituents
to which they are bound which enhancethcseroles.The study of the antifatty acids has shown that these substancescan even dictate, by themselves,difterentfates for the variousfatty acids they bind.
The combination of glycerol with any fatty acid seemsto establisha
caloric metabolic character. This is true for the very different fatty acids
found in plants and animalsas triglycerides.Even the ricinoleic triglyceride,
if fresh, is used as comestibleoil, castor oil. The same is true of the oil
of triglyceridesof polyunsaturated
fatty acids found in marine animal oils.
In the seeds,all the triglyceridesof fatty acids,even the conjugatedones
such as eleostearicand parinaric,representenergeticsources.It seemsthat
it is their combination with glycerol which has given all these fatty acids
value as caloric metabolites.The same is true for the bond to glycerophosphoric ion.
Combination with glycerophosphoricacid endows various fatty acids
with the abi.lityto participatein the constructionof membranes.The bond
to sterols,on the contrary, inducesan ultimate functional activity provided
the fatty acid iself is so constitutedas to be able to fulfill this function.
The influenceexcrtedby anti-fattyacidscan be understoodin terms of
the changesthey induce in the activity of the fatty acids.Sincethe activity
of the last is largely relatedto their presenceas free substances,
it is principally through their combinationwith fatty acids that the anti-fatty acids
intervene.By inactivatingthose free fatty acids which form a membrane
and insure its permeability,an anti-fatty-acidagcnt can causethe membrane to changeits permeabilityand evento becomecompletelyimpermeable. Similarly, an anti-fatty acid, by combining with a polyunsaturated
fatty acid, can reduce or even suppressits functional activity. It is to be
notedthat,by both changingpermeabilityand suppressing
functionalactivity, the anti-fatty acids exert their influenceultimately by altering oxygen
metabolism.From this point of view, metabolismbecomespredominantly
anoxybioticin contrast to normal oxybiotic metabolism.For glucose.for
instance,suppression
of the oxidativephasearrestsmetabolismat pyruvic
acid which passesinto lactic acid. The appearanceof acid substancesas a
biological effect of the action of anti-fafty acids results,in fact. from the
r.rPrDs^ND LrPotDS
137
Steroids
A fundamentalrole of thesesubstances
in biology is determinedby the
'l'his
fact that they are polycyclic.
leadsus to considerthe role of the ring
iself in reactivity, as shown by a study of the steroidsin oppositionto the
fany acids.In thc fatty acids.the bondsbetwecncarbonsas presentin the
aliphatic chain, insure a high reciprocalmobility between these carbons.
As a result, the entirealiphaticchain is highly llexible.on the other hand,
ri-uidityis characteristicfor all the rings,and is increascdby the polycycling
of the nrolcculcs.The constitucnts
of the moleculesare kept in fixed reciprocalpositions.while, in the fatty acids,the flexibility of the chain permits
the energeticcentersto take different relative positionsamong themselves
towardother nrolecule-s,
the rigidity o[ thc p<llycyclicmoleculcsmaintains
theenergeticcentersof the cycle,or thoseattachedto it, in the samerelative position. This fundamentalcharacteristic
of the cyclic moleculesappearsto be an important factor in determiningthe biological role of the
variousagentswhich have such cyclesin their molecules.
In the case of steroids,this attributcacquiresspecialimportance.An
understanding
of the differentbrologicalactiviticsof steroidscan be obtainedby an ana.lysisof the forces resultingfrom this characteristiccomposition.Besides the energeticcentersor formations attachcd to it, two
energeticcenters appear as part of the steroid nucleus itself. One is at
Cr and the other center is rcpresentedby the cyclopentanicgroup. The
factthat these centersare maintaincdin fixed relativeposition through the
rigidityof this polycyclicnucleushas resultedin an importantpropertyof
thenucleus itself which becomestranslatedinto a dipolarity of the molecule.The study of thesetwo energeticcentershas advancedour knowledge
of the role of steroids.
The study of the polar groupsbound to C,, of the polycycleskeletonof
138
nESEARcH rN pHysropATHoLocy
(a)
(b)
(L'
F l c . 6 4 . I n f l u e n c ee x e r t e du p o n t h e o x y g e n b o n d a t C ' b y t h e p o s i t i o n o f t h e d o u h l e
bond in the cycles I and 2 of the cyclopentanephenanthrene
molecule. A parallelism
b e t w e e nt h e d o u b l e b o n d o f o x y g e na n d t h a t p r e s e n tb e t w e e nC , a n d C , i n c r e a s e st h e
e n e r g e t i cc h a r a c t e ro f t h e c a r b o n y l ( a ) . A s i m i l a r i n f l u e n c eb u t l e s s a c t i v e , i s e x e r t e d b y t h e d o u b l e b o n d b e t w c e nG a n d C , . A d o u b l e b o n d a d d e d b e t w e e nC , a n d
C ' ( c ) i n c r e a s e st h e a c t i v i t y . S t i l l m o r e a c t i v i t y w o u l d r e s u l t f r o m a t b i r d d o u b l e
b o n d a d d e d b e t w e e nC c a n d C , ( d ) .
r*
,\/-t-r*oAA
(a)
rff
-k
.*
Ho,/W
*o&
(b)
(c)
6)
F t c . 6 5 . T h e i n f l u c n c ee x e r t e du p o n t h e h y d r o x y l b o n d a t C . b y a d o u b l e b o n d i n
t h e c y c l e I a n d 2 o f t h e p h e n a n t h r e n ei s i n c r e a s e di f t h e d o u b l e b o n d i s a d j a c e n to r
parallelto the bonds of C., bearingthe hydroxyl.
LTPtDSAND LrPorDS
139
140 /
R E S E A R c Hr N p H y s r o p A T H o L o c y
Crz also can have an oxygen or hydroxyl group and thus be negative or
positive. An important factor for the properties of the substanceis the
relationship between the two polar groups in the same molecule. It is apparent that the polarity of the molecule will vary according to what polar
groups are present at C3 and C17.[n a very simplified concept, which we
consider only partially accurate, we have tried to associate female and
male hormonal charact,eristicswith this polarity.
In the simple steroid molecules,a folliculinoid or estrogenicbiological
property seems to be conferred if the two polar groups-at Cs and Crz
-are formed by hydroxyls. The molecule appears to have a dipositive
polarity. It seemsto be important that the two hydroxyls be kept in the
relatively fixed reciprocal positions---+orrespondingto C3 and C,7-as
part of the solid skeleton of the steroids.Estrogenic properties are present
in various steroids that fulfill this condition. Furtherrnore, substancesfar
removed from the steroids have folliculinoid properties if they have this
relationship between the two hydroxyls. As shown in Figure 66 diethylstilbestrol,which has its two hydroxyls maintained in a fixed relative position similar to that of steroid estrogens, also shows potent estrogenic
activity.
OH
Estradiol
D i e t hIvs ti l b e s t r o l
Fro. 66. The lolliculinoid activity appears to be related to the existence of ru,o
hydroxyls kept at lhe same relative position. as it appearsin estradiol and in diethylstilbestrol.
ln the same way, we tried to correlate testoid activity with the presencc
of positive-negative
polarity; that is, with two polar centersenergetically
different, one corresponding to an oxygen and the other to a hydroxyl,
maintained in the same fixed relative position. The importance of this
relative steric position of the two polar groups for testoid activity becomes
evidentwhen it is found in substances
other than testosterone,
the principal
male hormone,with an oxygenat C3 and a hydroxyl at Cri. Testoid activity is presentin androsterone,which has an oxygenand an hydroxyl maintained in the samereciprocalpositions,althoughhere the oxygen is at C17
LIPTDS AND
LtPotDS
l4l
Testosterone
Androsterone
Ftc. 67. The testoid activity seemsto be related to the presenceof a hydroxyt and a
carbonyl in the same fixed relative position which is insured by tbe rigidity of the
steroid molecule. The same positional reciprocal relationship is seen to exist between
these two polar groups in testosteroneand androsterone.
The testoid activity secn for cortisone, hydrocortisone and other hormones also can be explained by the presencein these moleculesof oxygen
and hydroxyl at C3 and C17,and maintenanceof the fixed position between
thesetwo polar groups.
Conceprually,the antagonismbetweenestrogenicand testoid biological
activitiescan be consideredto be ultimately related to the differencesin
polarity, which in one form or anothercan be found in other factors differing for the sexes.we will men[ion here only that a similar differencebetween male and femalecharacteris seenin the sexualchromosomes,where
the female characteris related to the XX chromosome,and the male to
an X and a Y chromosome.As we will see below, a relationshipexists
betweenlipids in generaland sex.
Besidesthe sex hormones,fatty acids appear to be connectedwith male
sex characteristics,
while femalecharacteristics
are relatedto anotherg.oup
of steroids,the sterols.
Sterols
Characteristic of the structure of this group of steroids is the presencc
of a hydroxyl at Cs and a long chain bond at C,z. Through the hydroxyl,
:-ttr I
t.t
t;
142 /
xESEARcH rN pHysropATHoLocy
LTPTDS AND
LTPOTDS
143
betweenthe polar groups,the two coupledmoleculescan regain their independence.This would explain the relativelability of the combinationsberween fatty acids and sterols. The antagonism between fatty acids and
sterols is an important aspectof biological dualism which will be discussed
in more detail later when these substancesare studied in terms of their
influence at the difterent levels of organization.
144 /
nESEARcHtN pHystopATHoLocy
Corticoids
The corticoidsrepresentthe group of hormonesupon which the attention of scientistsrecentlyhas beenintensivelyfocusedbecauseof their new
therapeutic applications.
chemically, they appear to be the same as luteoids,derivativesof the
s:rmeparent hydrocarbon, allopregnane.Structurally, all these adrenocorticoid hormoneshave: a) a c3 bindingan o group; b) a double bond betweencr and cc in the first cycle;c) a two-carbontail chain with an o
attachedin ketone form to Czui d) an oH as primary alcohol presentat
c2t. This structure,common to all corticoids,seemsto be responsible
for
the principal propertiesof thesesubstances.
Corticoidshave been separated
into subgroupsbasedupon the presenceof aftachedgroups oH or _--o at
Crr or OH at Ctz. The presenceor absenceof attachedradicalsat C,,
appearsto be most important. Corticoidswithout attachedradicals at the
C1l have a major influenceon the metabolismof electrolytes.The second
group of corticoids,having the radical, are known as neoglucogeniccorticoids,the nameindicatingtheirprincipalbiologicalcharacteristics.
Energetically,the corticoids present a nucleophilic center at c3,
reinforcedby the presenceof the double bond in the cycle betweenCr and
cr. The doublebond is parallelto the doublebond of the carboxyl,and thus
inductivelyincreasesthe ionic characterof the latter.
A secondenergeticgroup of the tail chain appearsin toto as a strong
tripolarcenterwith a nucleophiliccenterat C.:,of this chain and an electrophilic center at c31. (Note 14) To this basic pattern is added, in the
neoglucogenic
corticoid,a separateenergeticcenter at C11.which can be
either clectrophilic,formed by a hydroxyl,or nucleophilic,formed by an
oxygen.
Corticoidsappeil, in general,to act as positivelipoids.(Note t5)
Becauseof their importancein relation to anti-fatty acid activity, we
will discussfirst the neoglucogeniccorticoids,the members with a polar
group also at C11.Accordingto our hypothesis,
thesesteroidshave a special biological activity, a role in the processof synthesisin the organism.
The part of the molecule between C11 and Crr constitutesan energetic
formation with a peculiar property. It representsa kind of energeticmold
or template,in which each carbon has its specificenergeticcharacter.Different radicalswould be attractedby the energeticcentersof this template
formation accordingto their own energeticnature.Kept in their respective
positions,they would be induced to bind together in order to fornr new
substances.In this manner this template formation would promote new
L T P T D SA N D L T P O T D S /
145
146 /
R E s E A R c Hr N p H y s r o p A T H o L o c y
LIPIDS AND
LIPOIDS
147
148 /
nEsEARcH tN pHysropATHoLocy
in an exaggeratedmanner to oppose the fatry acids physically and chemically, thc same means can be utilized to reduce this exaggeration.Thus, the
intervention of glucuronic acid as a result of an abnormal oxidation of
glucooe induced by fatty acids appears to be biologically sound. This is
also true for the sulfuric radical.
The importanceof thesesubstancesdoesnot resideonly in the fact that
the organism can easily produce them in larger quantities than fatty acids,
The fac-tthat they combine to form excrementalsubstancesis important
too, for in this way, they help in materially eliminating the anti-fatty acid
substancesfrom the organism. This would not take place if only a combination with fatty acids were possible,since the esters of fatty acids are
usually rotained in the organism and, under certain circumstances,can
again liberate their constituents.The intervention of glucuronic acid and
sulfuric acid appearsto be more effeotivethan that of the fatty acids which
have their own activity and are more toxic in exaggeratedamounts.This
appears to be especially true in the case of glucuronic acid becausethe
amount of glucose available is practically unlimited as compared with
other metabolites.Glucuronic and sulfuric acid would thus intervene in
the biological antagonism botween fatty acids and anti-fatty acid substances,inactivating and eliminating agontsfrom the last group, especialty
when in excess.Teleologically speaking,their intervention appears to be
still more interestingsince the body has, as part of its defensemechanism,
a tendencyto manufactureanti-fatty acids in excess.The interventionof
agentsother than fatty acids would prevent a vicious circle and permit an
excessof anti,fatty acidsto be removedby excretion.
FATTY ACIDS VS ANTI.FATTY
ACIDS
This study of the relationship betweenfatty acids and anti-fatty acids has
been guided by the dualistic concept. It must be recognized,however, that
the direct activity of thesesubstances
could be largely reducedto that of
one 8roup, the fatty acids. The action of anti-fatty acids is largely indirect.
They control and thus limit the activity of the fatty acids. It is within this
framework that the different anti-fatty acids selectivelyinfluencedifferent
specificfunctions of the free fatty acids.
The lipids and asscciated constituents with their multiple activities
create for each entity a balance responsiblefor many of the manifestations
of the entity. Variations in manifestationscan be attributed in large part to
qualitative and quantitative variations in the intervening lipids. A sys-
:,,;'r)
LTPTDSAND
LTPOTDS
t49
150 /
x E S E A R c Hr N P H Y s r o p A T H o L o o y
Age
The changesin lipidic balancerelated to agehave been made the object
of an extensivestudy which also sought to determine the role of lipids in
aglng processes.A general predominanceof positive lipids, more manilest
in the cellular and tissue levels than in the blood, was seen in youth. This
would be expected in view of the special motabolic influence exerted by
this group of lipids. The anoxybiotic charaoter of metabolism induced by
sterolsresultsin the intervention of dehydrogenases
which lead to an abundance of hydrogen ions. This, in turn, leads to a predomhance of the kind
of syntheseswhich favor anabolism. Growth thus could be related to the
predominanceof lipids with positive polar groups, especiallysterols.
Agng processes,on the conEary, could be related to a predominance
of lipids with negativo polar groups, especiallyfatty acids. This predominance could be found especiallyat the cellular level, as seen in cultures of
tetrahymena.(Note 23) ln complex organismsor in rats (Note 24) in which
an increasein the proportion of fatty acids at the cellular level is present,
an opposite change occurs at the systemic and even at the organic level.
There is an excessof cholesterol,this time limited to the higher levels, as
revealedthrough analysesof the blood, for instance.Changesin the blood
vesselsare related in part to this excessof sterols at the systemic level.
Many manifestationshave confirmed such an offbalance with sterol predominanceat higher levels.For example,we found the urine surfacetension
abnormally high in old age. (Note 25l Similarly, skin wheal absorption
in old people requires more than 90 minutes for completion as against
approximately 20 minutes in middle-agedaduls. A predominanceof fatty
acids at lower levels and of sterols at higher levels would thus characterizs
the changesin lipidic balancerelatedto old age.(Fig. 68)
O ther Physiological F actors
The study of the role of lipids in various physiological functions was
made indirectly for the most part, using the tests previously mentioned
which were interpreted in terms of dualistic patterns. These were related to
the general offbalances A and D and, through them, attributed ultimately
to a predominanceof sterols or fatty acids.
Sleep in itself, without relation to night or day, was found to induce
a marked change,comparableto a type A offbalancewith predominance
of sterols. Subjects with pain of an acid pattern often correlate the appearanceof pain with sleep, the pain occurring uniformly at the moment
they wake up. In these cases,the urine shows a low specific gravity with
L T P T D SA N D
LTPOTDS
l5l
a high pH and a high surface tension,correspondingto an A type offbalance.As we will see, in subjectswith an intensiveA type offbalance,
nocturnal polyuria and pollakiuria occur.
sexual intercoursein males was seento induce,in analyses,transitory
changessimilar to an offbalanceof type D, correspondingto a predominance of fatty acids.In femalesthe changecorrespondsto a transitory offbalanceof type A, manifestedby changesat the systemiclevel. Muscular
exercisewas seen to induce, in a first phase during the exerciseitself,
r00
too
60
/"
40
20
0 1 0 20 l0 10 50 60
minutes
S u b j e c t sb e t w e e n
15 and35 years
l020lo{o5060?oo
minutes
S u b j e c t sb e t w e e n
65 and87 years
F I c . 6 8 . T h e d i s a p p e a r a n ctei m e f o r t h e w h e a l i n d u c e d b y t h e i n t r a d e r m i c i n j e c t i o n
of 0.2 cc saline, varies with the age. In old age, the wheal oftcn persistsfor more
than 90 minutes.
152 /
REsEARcHrN pHysropATHoLocy
appear in a new light when interpreted not as being the direct results of
time changesbut rather of patterns of diurnal activity and nocturnal rest.
This explainswhy in rats and mice, which are nocturnal animals,most of
the analysesshow variations related to the time of day opposite to those
in humans. other variations could be recognizedmore strongly related to
time of day. variations with a 24-hour rhythm could be seen,for instance,
for urinary surfacetension in mice. But, when rats and mice were maintainedfor a length of time under artificialcondirions,with light during the
night and dark during the day, the animals changedtheir habits, becoming
active during the day and sleepingduring the night. Afrer a certain time,
most of their analytical patterns such as urinary pH, blood leucocytes,
eosinophilesetc. changed,acquiringthe type of variation seen in humans.
Urinary surface tension remained unchangedfor a long time. (Note 8
chapter IV) Even more interestingwere other changeswhich could be related to changes in externa.ltemperature.The urinary surface tension
measuredin rats in the morning for long periods of time showed variations relatedto changesin the temperatureof the environment.(Note 26)
(Fis.69)
The importanceof temperatureled to its more detailed investigation.
Variations in lipidic balancehave been found to para-llelvariations in body
temperature.The blood of normal individualsis richer in sterolsthan the
blood of those with hypothermia.Furthermore,in moments of high temperature,more sterolsare found than in momentsof low or normal temperatures.An increaseof fatty acidsoccursin conditionswith hypothermia.
These changeswere confirmed also by the correlation between blood content in lipids and temperaturein different abnormal conditions. In shock
with hypothermia the blood is rich in fatty acids, while in infections with
fever.it is rich in sterols.
The role of.temperaturewas also investigatedby studyingthe influence
upon the lipidic balance by externally applied heat or cold. Characteristic
variationscould be seenin human analysesunder the influenceof hot and
cold days, and of local applicationsof heat and cold. Manifestationscorrespondingto predominanceof lipids with positive characterwere induced
by heat, while others correspondingto predominanceof Iipids with negative characterwere inducedby cold. Variationswere seenin animalskepn
in an incubator or in a refrigerator. ( N ote 27) w e will seebelow, by studying their influence at different levels, the importance of these variations
produced by temperature.
The influence exerted by barometic change.scould be seen in changes
in total blood potassium,the two curves being paraUel.Similar changes
LTPTDSAND
LTPOtDS
153
t2
t,
q,
L
5t
.\t ao
(-
c,
o- 32
E
o,
6'
v,
(u
.U
z
v,
(a
s5
vt
(u
(t
E
(u
6t
l!
./,
q
0aYs
Ftc. 69. The curves of the average values of surface tension of two groups of 20
malc rats cach, and two groups of female rats each, show parallel changes with thc
inverse curve of the temperatureof the environment.
154 /
nESEARcHtN pHysropATHoLocy
L T P T D SA N D
LlPOlDs
155
156
REsEARcH rN pHystopATHoLooy
and unsaturatedacids and alcohols. Almost no activity was seen for the
saturated.The unsaturatedmemberswere active in general,with activity in
any direction increasing with the degree of desaturation of the nonpolar
group. Thus, it appears that the nonpolar group determines whether a
substanceis active or inactive, but the nature of the activity-that is, increasingor decreasingreceptivity-is determinedby the polar group.
The influence exerted by agents with a positive charact,er upon viral
infection would explain the seasonal changes in clinical manifestations
which are especially interesting for the paralytic form of poliomyelitis.
we could show experimentally that when mice, after being inoculated
subcutaneously with smallpox vaccine virus, are kept in an incubator at
37"c, all develop cerebral involvement, while such involvement appears
in only a small proportion of other animals kept at room temperature, and
does not appear at all in those kept in a cool place. As we could also show
that one of the effects of exposure of an animal to a higher temperature
is an increase in the body of the amount of free lipids with positive character, this could explain the increasein the virus sensitivity of cells in the
central nervous sys0emwhich are especiallysensitiveto these lipids. This
relationship would also explain the increasedincidence of paralytic polio
casesduring hot weather.
The presence of greater amount of lipids with positive character in
youth helps also to explain the frequency and intensiry of viral infections
in children.(Note 31)
The study of the effects of temperature and lipids upon viruses has
shown that those effects are not limited to the host but also are exerted
upon the viruses. (Note 32) The influence of heat and cold upon virus
activity was studied in bacteriophages,where effects for virus and host
could be separated.The direct influenceupon the virus appearedrelatively
small and secondaryto the changeswhich appear in the host itself. Bacteriophage,separatedfrom microbes by filtration and kept in an incubator at
temperatures 2-3 degrees C higher or lower than controls, showed no
change in virulence. This was true as long as microbes were not present.
Microbes kept at higber temperaturewere more sensitiveto phages;when
kept at lower temperature, they were less sensitive. This influence went
so far as to change a sensitivestrain to a refractory one, and vice-versa.
The fact that microbesgrown at higher temperaturesfavor the develop
ment of bacteriophagewhile those grown at lower temperatue hinder it
could be correlatedwith the changein the richnessof lipids in the microbes
themselves.Similar results were obtained when microbes were grown for
a time in media containing fatty acids or insaponifiablefractions and were
l;r:iS$r
i
LIPTDSAND LrPolDS
157
then removed and exposed to phages.These experiments(Note 3Jl indicate the direct role played by the lipids of the hosts in the activity of
phages,and would explain the influenceexerted by temperature.Through
the change in the lipids of the hosr, the virus changestoo, becoming more
active if grown in microbes at a higher temperature and less aggressiveif
passedthrough microbes kept at lower temperature.
Efrects ol Lipids on Miuobes
The antagonisticeffectsof the two groups of lipids upon microbes were
investigated. As an example, we will mention here the characteristic
changesin Bac. anthracis treated with polyunsaturatedfatty acids and insaponifiablefraction preparations.(Fig. 70) We investigatedthe microbes
for their morphological, tinctorial, cultural and virulence characteristics.
with the fatty acids added to media, changeswhich can be consideredto
be mutationalwere induced,leadingto tiny Gram negativemicrobesgrowing on agar as transparentsmall colonies.The changes,however, were
reversible.Usually several passagesin normal media were sufficient to
produce reversal.First small and separate,then larger and more confluent
Gram positive granules were seen to appear in the microbes which, themselves, also became progressivelyplumper. Ultimately, all the characteri51is5,-rnsrphological, tinctorial and cultural---of the normal microbes
reappeared
. (Fig. 7l )
Microbes showed opposite changes when treated with insaponifiable
fractions,(Fig. 70) losing their bacillusform. Abnormally intensiveGram
6
(a)
treated.
with
sterols
i=L
\-
(b)
(c)
t?:ttio.:ilh
controI
Fro.70. lnfluence ol lipids upon microbes. Schematic drawing of the changes in.
duced in Bacillus Anthracis by the influence exerted by the two groups of lipids.
Treatcd with sterols (a) as in tbe unsaponifiable fraction of placenta, the microbes
change into cocci irregularly shaped and intensely retaining the gram stain. Treated
with fatty acids (c) from cod liver oil, the bacilli change into very tiny gram negative microbes. (b) shows untreated microbes.
..,;
t.t;.'i,*i;;',;,.
."t.
""#.;-14
158
nEsEARcH rN pHysropATHoLocy
=/--'
- -
a{a*
e&
-{.'
\
positive cocci appeared. They grew on agar as very thick creamy white
colonies. These changeswere seen to persist for a long time and seldom
were sPontaneouslyreversed.Treatment with fatty acids induced reversal
although inconsistently. we attempted to correlate the differences in
changesinduced by different lipids to the different levels of the microbe
at which they work. The changeto cocci can be regarded as corresponding
to an influenceexerted upon the membraneand the change to Gram positive to an influence upon differentiated formations present in the body.
( 3 1 3)
Effects of Lipids on Protozoa
The effects of lipids upon monocellular organisms,especiallytetrahymena pyriformis, were studied and an effort made to relate the nature
of the main changesinduced in these protozoa to changesobserved at the
cellular level of complex organisms. An initial effect was noted on the
polarity in protozoa which seemed to be oppositely influenced by long
chain polyunsaturatedfatty acids and sterols.Lipids with a positive character were seen to induce a change in the form of protozoa causing them
to become almost round, a change considered to correspond to reduced
polarity. Lipids with a negativecharacterhad an oppositeeftect;the tetrahymenabecameabnormallyelongated.
The administration of higher amounts of polyunsaturated fatty acids
was seento induce immediate changeslocalized at the anterior pole of the
organism, changeswhich ultimately lead to the breakdown of the membrane particularly at this point. This effect parallels in intensity the degree
of desaturationof the fatty acids. other changeswere seen in growth rate
and survivxfrime and, thus,in the agingprocess.(Nole 3a) (Fig.7a)
At the same time, resistanceto heat was seen to increaseas the result
of treatment with negative tipids, while it decreasedafter treatment with
LIPIDS AND
ControI
LTPOIDS
r59
with
Treated
fatty acids
Ftc. 74. In a direct action of fatty acidr on tetrahynlcna, a pas:,agcof fluicl occurs
at the surface with a break of the membrane especiallymanifest at the anterior polc
( a ) , c o n t r o l u n r r e a t e d( b ) , ( 1 2 0 0 x ) .
160 /
R E S E A R c Hr N p H y s r o p A T H o L o c y
L T P T D SA N D L T P O t D S /
151
162 /
REsEARcH rN pHyslopATHoLooy
163
has a marked diuretic eftect which we will discuss below with its therapeutic aspect.
Nervous system-rnteresting eftects by the two groups of antagonistic
lipids upon many manifestationsof the central nervous system have been
noted.
Convulsions-Administration of sterols and insaponifiablefractions of
many organs such as placenta, liver, butter, eggs, etc., in large amounts
induces convulsionsin rats. Convulsionsalso were noted in humans when
huge dosesof these agentswere administered.But even in relatively small
amounts,theselipid agentssensitizedanimalsto the administration of other
c-onvulsantagents.In rats or mice receiving such lipids, thiamine chloride
induced convulsionsin doseswithout effect in controls. (Note 39)
An opposite effect was observed for lipids with negative character.
Saturatedfatty acids showedno influenceon thiamine-inducedconvulsions.
Such convulsions were prevented by the administration of nonsaturated
members.The effect was related to the degreeof desaturation of the fatty
acids. with increasesof the iodine number, the necessaryeffective doses
of these fatty acids becameprogressivelysmaller. While hundreds of milligrams of mono- and diethenicacids were necessaryfor each 100 gram of
body weight, the anti-convulsanteffect was obtained with only a few milliSams of clupanodonicacids,and with still lessof the nonenicacid, bixine.
The study of the pathogenesisof convulsionsalso covered the influence
exerted by these lipids of the adrenal corticoids. The administrationof
mineralocorticoids,especiallydesoxycorticosterol,
even in small doses,to
who
subjects
had receivedany one of the lipids with a positive polar group,
such as cholest,erolor insaponifiablefraction of placenta,liver or kidney,
was followed almost invariably by convulsions.we will presentmore details on this effect later in the discussionof syntheticsubstances.For the
moment we want only to note the relationship between mineralocorticoids
and lipids with positive character in the pathogenesisof convulsions.The
concomitant intervention of the two factors-an oftbalance induced by
lipids with positive character, and action of mineralcorticoids-seems to
provide new light on the pathogenic problem of epilepsy and convulsions
in general.
Coma-The role of cortical hormonesin the pathogenesisof convulsions was confirmed by the opposite effect produced by neoglucogenic
corticoids. We will see later that the administration of cortisone to subjects
receiving higher alcohols such as heptanol, octanol or octandiol in large
doses,induced a subcomatosecondition at first which progressivelychanged
into coma. (Note a0) Oppositepropertiesof the mineral and neoglucogenic
164 /
REsEARcH tN pHysropATHoLocy
165
rirt';:l;
l.';.rr
166
RESEARCH IN
PHYSIOPATHOLOGY
EFFECTS OF STEROLS
Cells
Prolongsyouth character
Increases
potassiumcontent
Decreasessodium content
Reducesmembranepermeability
Reducescellularoxidation
Reduceschloridecontent
Inducesrapid aging
Decreasespotassiumcontent
Increases
sodiumcontent
I ncreasesmembranepermeability
I ncreasescellular oxidation
Increaseschloride content
Tnissaes
Lowers pH of lesions
Lowers chloride content of
lesions
Lowers water content of lesions
RaisespH of lesions
Raiseschloride content of
lesions
Raiseswatercontentof Iesions
Organs
Inducessomnolence
Inducesdiuresis
Inducesconstipation
lnducestachycardia
Inducesinsomnia
Inducesoliguria
I n d u c e sd i a r r h e a
lnducesbradvcardia
Systemic Induceshyperthermia
Induceshypertension
Blood
Increases
RC volume
DecreasesRC sed.rate
Increasespersistenceof
oxygen fixation
Determinespersistence
of
RC isolation
Determineshyperleucocytosis
Determineseosinophilia
Decreaseskalemia
Induceshypothermia
Induceshypotension
DecreasesRC volume
IncreasesRC sed. rate
Decreasespersistenceof
oxySenfixation
Determinesformation of sludge
Determinesleucopenia
Determineseosinopenia
kalemia
Increases
Induceswater retention
I nducessulfhydryl excretion
lnducescalciumretention
I nduceschloride retention
Inducessodiumretention
Inducesphosphateexcretion
Inducesexcretionof surface
activesubstances
167
168 /
nEsEARcH rN pHysropATHoLocy
..rr,.,,fra.
$tii.iti:t:r,:.;!$11;li:r1
169
Amino Acids
Amino acids have been separatedinto groups basedupon their effects
at different levels. The first group includes the simple amino acids. In
these members the portions of the molecule which are added to [he amphoteric amino acid group, are usually electrically neutral. The amino acids
polymerizedthrough the anaphotericgroup serve as building materialsfor
the bigger protein molecules. They have appeared to be inert without
effects upon the different levels. Beyond these simple amino acids, are
two groups, energetically active, which have a second energetic center
with a negative or positive character in their molecules. while the amino
acid group servesto make thesesubstancespars of higher proteins through
the same bonds of amino acid groups as the simple members, it is the
other energeticcenter, with acid or alkaline character, which confers upon
these amino acids a positive or negativecharacter.
We studiedeffects,at difterent levels,of arginine,lysine and histidine,
which are members of the group with alkaline centers; of glutamic and
aspartic acids which have acid centers; and of methionine with a thiolic
center. Like for the lipids, the last two groups have shown similar propertias,but oppositeto thoseof the memberswith dkaline centers.The nature of their intervention appearedevident through the interesting opposite
effectsexertedupon microbes.Cultures of B. subtilis in broth containing
membersof one or the other of the antagonisticgroups show characteristic
changes.Unlike controls in which the long chains of microbes remain isolated, the microbes were seen to be kept together in media with atkaline
amino acids, forming a consistent gelatinous mass separated from the
medium. In broth with acidic or thiolic amino acids, the microbes remained separatedor formed very small aggregates.This appearedinteresting when we considered the positive character present in a.lkaline amino
acids, as related to the heterotropic,constructivetrend, while the negative,
as in the acid and thiolic members,is related to the opposite trend, We saw
further the sameantagonism betweenthe influenceexerted by histonesand
nucleic acids, the first paralleling the alkaline amino acid groups and the
second the opposite group. The more manifest effect of the ribonucleic
acids could be seen to take place at higher levels of the organization and
possibly explains the more direct action upon the genes.
We investigated the eftect of the two groups of amino acids at the
tissuelevel upon pain. Arginine,lysineand histidinedisplayedan analgesic
effect upon alkaline pain, while glutamic acid and methionine had this
effect upon acid pain. The effect could be related more to the basic tend-
#4i#*r;.i;
..,..b*
170
xESEARcH rN pHysropATHoLocy
CHAPTER
DEFENSE
rTr\
REcocNtrtoN THAT multiple factors are responsiblefor abnormal
I
"t
conditions,
and that these factors can be systematizedaccording to the
concepts presented above, throws new light on a specific aspect of the
relationship between the different entities and the environment when this
tends to alter their characteristicorganization.This responseis concretized
as the defenseagainstthe noxious.The analysisof this defensehas been
facilitatedby emphasizingthe relativeindependence
of the endtiesforming
the complex hierarchic organism,the dualistic patterns of response,and
the critical role of the lipids as well as of proteins. Abnormal processesin
an organism's defense system may be better understood when they are
compared to those correspondingto normal physiologicalprocesses.For
this reason,we start with this last aspect.
The direct intervention of a noxious agent upon a biological entity can
be characterizedby its tendencyto induce heterogenization,
through an alterationof the entity'sconstituents
or the relationship
betweenthem.This, in
turn, affectsone or more of the constantsthat characterizn,the entity. The
ensuing defense response is directed ultimately at restoring the altered
constantsto their normal values.
Involved in a first stage of defenseare those very factors which normally maintain the constants,the factors which induce the oscillatory
dynamic balance. As a first response, they become exaggerated. Such
exaggeration,which takes place successively
for the opposite phases,resolves many slight noxious interventionswithout clinical manifestations.
Through a damping movement, the exaggeratedoscillationssoon return
to normal. If the normal constantsare reestablished,
the phenomenoncan
be consideredto be a physiologicalresponse.
But if the alterationsinducedby the noxiousagentpersist,an abnormal
L7r
172 /
RESEARcHIN pHystopATHoLocy
condition results. Indeed, in this casethe exaggerationof oscillatory movement can be so great that an abnormaliry may result even from this exaggeratedattempt of the entity to reestablishnormalcy. In fact, offbalances
are induced by just such changeswhich often represent,by themselvesas
will be secnlater, one of the major immediatefactors inducing the abnormalcy. As long as an abnormal condition is not resolved,the biological
entity will try to utilize new meansin order to reestablishthe normal balance. If the constantsdisturbed by the noxious intervention are fundamental, or if the changesresultingfrom the defensemechanismitself are
too great, death of the entity will result.
As expected,responseswill differ accordingto the level to which an
affectedentity belongs.However, despitethe many differencesrelated to
levels, a common and relatively simple pattern can be recognizedwhen
manifestationsoccurring at different levels as tlre result of the noxious
intervention are compared and referred to the basic pathological concepts
already noted.
Most of the information about this simple pattern was originally obtained by srudyingresponsesat the systemiclevel. Blood was parricularly
suitablebecauseof its availability,its multiple constantsand manifestcapacity to conserve them, and particularly because of the facility with
which noxious agentscould be inducedto act upon it.
The interventionof a noxious agent able to change the energeticbalance of blood sets in motion immediatelya group of successiveprocesses
which may or may not be clinically apparent,dependingupon their intensity.They have been describedas hemoclasiaby widal and hemo-shock
by many authon. Although widely investiga,ted,
the mechanismdid not
appear clcar. From our studies, we have arrived at certain conclusions
which we will briefly prasenthere.
Diphasic Phenomenon
As a noxiousfactor, we usedan intravenousinjectionof killed microbes
or of a colloidal suspensionof a metal. within a few minutes, a group of
changesoccurred.They were revealedthrough a seriesof analysesmade at
very short intervals.(Note /) The changeswere found to affect most of
the blood constituents,The most characteristicchangein our opinion is a
leucopeniawhich especiallyaffects the granulocytes.With it, there is a
lowering of serum antitryptic power; a decreaseof serum albumin; appearance of degradatedproteins,esteraseand amylase;increaseof free fatty
acids; and a lowering of coagulabilitywith reduced clot retraction.Clinically, these changesare accompaniedby hypothermia and hypotension.
DEFENSE
173
Together they representwhat we will call the "negative phase" of the immediate response.
This group of changesrepresents,in fact, only the first part of a diphasic phenomenon.The negativephase is usually followed by a second
and opposite one which we call the "positive phase" of this immcdiate
response.It results from the tendencyof the body to correct, and even
over{orrect, the changesoccurring in the fint phase. After hypothermia
and hypotension,hyperthermiaand slight hypertensionfollow. At the same
time, thc number of granulocytesincreases,as does the antitryptic power
of serum and its albumin contcnt. The serum appears richcr in frec
stcrols. Blood coagulabilityand clot retractionalso increase.After moving
rapidly to a peak, all thesevaluesreturn slowly to normal. The existence
/eo
(u
+t
rJ
o
Zeoo
(J
:t
q,
J
l'{inutes
Frc;, 75. Diphusic responsein the delense.The intravenousinjections to a normal individual of a foreign material such as of a suspensionof killed microbes or of a colloidal
metal induces a typical responsewhich correspondsto the hemoshock. A diphasic
curve seen in most of the analysescharacterizesthe occurring variations. The curve
presentedcorrespondsto the total number of the blood leucocytes.A parallel diphasic
curve is seen for other blood analysessuch as clot retraction, albumin content of the
s e r u m , a n d a n t i t r y p t i c v a l u e s o f t h e s e r u m . S i m i l a r d i p h a s i cc u r v e s , b u t o p p o s i t e i n
sens,are seen for blood coagulation time, amount of arnylase and esterasein the
\ e r u m . a m o u n t o f K , i n t h e s e r u m , a n d f o r t h e a m o u n t s o f p r o t e o s e sa n d p e p t o n e s
#t!*
idi\
tthi.l
ilF
, . tt."
.,.
174 /
xEsEARcHrN pHysropATHoLocy
of two phases can be recognized in all the changes occurring in hemos h o c k(. F i g . 7 5 )
In trying to correlatethe multiple changestaking place, it is the lysis
of leucocytes,especiallygranulocytes,which can be considcredo[ primary
importancein the developmentof hemo-shock.This is evident from the
relationship between granulocytopeniaand the intensity of the diphasic
phenomenon.The administrationof morphine or other opium derivatives
to an individual,prior to the applicationof the noxious factor, will reduce
or suppressthe granulocytopeniatogether with all the manifestations.
(Note 2/ Intensive physical exerciseconcomitantwith the application of
the noxious factor will increasethe granulocytopeniaparallel with all the
manifestationsof hemo-shock.(Note 3)
According to our hypothesis,lysis leads to liberation of proteolytic
enzymeswhich may be present as such or may be present in precursor
form in the leucocytes.And it is the interventionof theseenzymeswhich
reducesthe antitryptic power of the blood and, by digestingblood constituents,Iowers the amount of albumin presentin the serum, and induces
a parallel increaseof productsof protcin hydrolysis.The increasein anrylase as well as in esterasepresentin blood is related to the other hydrolytic enzymesliberated in this phase,and is also probably correlatedwith
leucolysis.The esterascacts hydrolytically upon the neutral fats present
and this would explain, at least in part, the liberation of free fatty acids
seenin this phase.In the changescorrespondingto the first phase,digestive
effect of theseenzymesupon the blood constituentscan be recognizedas
being one of the most important interveningfactors.
We confirmedthe correlationbetwcenthesechangesand leucolysisnot
only through thcir parallel variations, as mentioned above, but also
through in-vitro experiments.Lysis of leucocytesresultedin liberation of
hydrolytic enzymcs.An exudatc rich in granulocyteswas obtained by
injectingsterilebroth, or an aleuronsuspension,
into the pleura of rabbits.
To this exudate,removed through pleural puncture, a small amount of a
colloidalsilver-proteinpreparation(Collargol0.lvo) was added and the
preparationmaintainedat 38oC. This was seento induce the appearance
of vacuolesin the leucocytes,following the phagocytosisof silver grains.
The vacuoleswere observedto grow rapidly to huge dimensionsfollowed
(FiS. 76)
by burstingof the leucocytes.
Analysisof the pleuralfluid treatedin this mannerhas shown the same
changeas those seenin the first phaseof hemo-shock:loweringof antitryptic power with a decreasein albumin content, increasein products
formed by partial digestionof proteins,appearanceof amylaseand ester-
DEFENSE
l7S
ase, and an increaseof free fatty acids. There were also the same nuclear
"shadows" as encounteredin large amounts in the circulating blood at this
phase. The increaseof the potassiumcontent of serum seen in this phase,
and the increase found also in the supernatantpart following centrifugation of the exudate to which Collargol had been added, representsa further
confirmation of the role of leucolysisin this first phase.These data enabled
us to consider that the mechanismthrough which the blood tries to combat the intervention of a noxious agent corresponds,in the first phase,
primarily to a lysis of granulocytesfollowed by hydrolytic digestion.
.%g
lrorrr Granrlocytes
"P @lii,,lr.anu,es
@
" "/*crear
shador
Leucocytes
traated
rtth Collarjol
FIc. 76. Drawing of the changesinducedby a colloidul su.spension
ol silver proteinate
upon leucocyles.The leucocyteswerc obtained by injecting broth intrapleurally to
rabbits. Silver proteinate was added to thc suspensionof leucocytesand thc changes
observed in a microscope heated chamber maintained at 38'C. The phagocytosisof
the silver proteinate leads first to the appearanceof this substanceas intracellular
granules, followed by the formation of vacuoles.As these grow to a huge size the
cells burst. The nucleus remains as nuclear shadow.
176 /
nESEARcHrN pHysropATHoLocy
If this hemeshock, in spite of its frequently violent clinical manilestations, resolves the effects of the noxious intervention upon the blood, it
can be considered to be, to a certain degree, a physiological response.It
amounts to an exaggoration of the oscillatory mechanism through which
the characteristicconstantsof the blood are maintained. By employing the
hydrolytic enzymesstored h the leucocytes,the blood tries to resolve the
influence exercised by the noxious factor, digesting and thus breaking
down either the factor itself or the results of its direct intervention. Acting
upon blood constituents,the noxious agent often inducesthe appearanceof
micelles bigger than those normally circulating. The fatty acids liberated
by hydrolytic enzymes would insure, in the first place, a higher boundary
permeability, thus permitting the passagethrough the capillaries of substancesotherwisebarred. At the sametime the fatty acids bind the antigen
in a lipidic complex.
In the second phase, the organism tries to repair damagescaused by
the exaggerateddigestive processor by the intervention of fatty acids. If
the organism is able to resolve through a successfuldiphasic reaction the
changesinduced by the noxious agent,it returns to normal.
Prolonged Hemoshock
Inability of the organism to resolvethe noxious interventionthrough
the mechanisminvolved in the diphasic phenomenonleads to abnormal
prolongationof one phaseor another.If it is unableto destroythe noxious
factor in the first phase or to mobilize the repair process in the second
and thus correct the damage induced by the first phase, the organism remains in a prolonged first phase of hemo-shock.If the second phase is
quantitativelyor especiallyqualitativelyinadequate,the organism remains
in a prolongedsecondphase,continuingto try to resolvethe offbalanceby
a quantitativelygreater mobilization of the otherwisequalitatively inadequate weaponswhich are at its disposal.It is the predominantintervention
of the lipids which characterizestheseextendedphases.We wish to note
again that the fatty acids intervenein the prolongedfirst phasewhile antifatty acid agents,especiallysterols,are active in the second.
The adrenalsplay a particularly important role in the immediate and
prolonged defense process.In the first phase, the increasedamount of
fatty acids with four or morc double bonds found in blood and in the
organismin generalappearsto conrefrom the adrenals,which are usually
extremelyrich in thesesubstances.
In an exaggeratedlyintensiveprolonged
first phase,we found small reddish adrenalspracticallydevoid of fatty
acids.This occurrence,togetherwith the concurrentincreasein fatty acids
DEFENSE
177
in the blood, relates thesechangesin fatty acid content of the blood largely
to a Iiberation of the adrenal fatty acids into the circulation. Another important factor for the prolonged first phase appears in the intervention of
lymphocytes able to induce a lysis of compounds of very high fatty acids
such as presenteven in waxes.(Note 4) A lymphopeniacorrespondsto the
prolonged first phase. In the elevation of the amount of anti-fatty acid
agents in blood, characteristic of the prolonged second phase of the diphasic phenomenon, the adrenals seem to intervene again providing a
portion of the increasedcirculating sterols. The exaggeratedmanufacture
of sterols can be attributed to the reticuloendothelial system in general.
cranulocytosis and lymphocytosis occur in this prolonged second phase.
The interventionof sterols, which are relatively simple steroids,can explain the clinical manifestationssuch as fever, which charactcrizethe prolonged second phase, since fever can be induced by the administration of
Iarge amounts of sterols.
we can separate, from the point of view of its manifestations, the
immediatediphasichemo-shockphenomenonwith a short evolution, from
the more prolonged forms. while the former, if not too exaggerated,would
correspondto a physiologicalphenomenon,t}tc latter is always abnormal.
In the former, the principal interventionis that of hydrolytic enzymes;in
the latter, lipids play the most predominant role. pathogenically,each
phase of the diphasic phenomenon,if unable by itself to resolvethe immediateproblem, will be followed by a correspondinglipidic predominance.
The result may be either one of the two phascs,with fatty acids or sterols
predominant.We call this entire response"the antiheterogeneous
reaction"
of the defense,separatingits diphasic manifesrationsinto immediate hydrolytic and prolonged lipidic stages.
A rui heterogeneous Reaction
Although, in the prolongedlipidic stage,a certain specificiryfor particular antigens can be recognized,the antiheterogeneousresponse in
generalrepresentsrather a nonspecificeffort of the organismto resolvethe
problemscausedby the presencof any heterogeneous
factors as such.
Before going further, we want to cmphasizesomc important characteristics of this antiheterogeneousresponse related to organization. The
catabolic processespresent in the first phasc appear to result in part from
the direct hydrolytic process and in part from the biological intervention
of the products of hydrolysis, especiallyfatty acids. The hydrolytic enzymatic processis homotropic in nature by delinition, as it breaks down
difterent constituents,liberatinggroupswith anionic and cationiccharacter.
178
n EsEARcH rN pHysropATHoLocy
DEFENSE
179
tain degreeof specificity toward the antigen. Two kinds of antibodies will
be manufactured and will differ in their fundamental characteristics,the
time of their appearance,and their role in the defenseprocesses.
CoagulantAntibodies
The first group of antibodies have a characteristicproperty. Together
with the antigen, on which they fix with a degree of specificity, they form
highly energeticcomplexes.This is manifest in a marked tendency to bind
together such complexesas well as constituentsof the blood and form huge
aggregates.When such antibodies are produced for, and act against, a
specific microbe, agglutination results. Conglutination, prccipitation and
flocculation occur when similar antibodiesact against other antigens.Due
to their tendency to establishantigen antibody complexesresulting in huge
formations, these antibodiesare generallygrouped as coagulant antibodies.
Although the coagulation characteristic is not demonstrable in vitro for
all antibodies in this group, we use the term "coagulant antibodies" for
didactic purposes.
The huge complex formation resultingfrom the binding of coagulant
antibodieswith an antigencan appear as a precipitate,agglutinatedmicrobe
or conglutinated red cells. Once established,this formation representsa
new heterogeneousentity of much larger dimensions than the antigen
alone. As srrch,it becomesby itself a new noxious agent for the organism
which consequentlyreacts against it. The organism utilizes the same processesagainst this noxious antigen-coagulantantibody complex as it uses
for any heterogeneousagent, with the same immediate diphasic or prolonged mechanism.
Teleologically,the formation of coagulantantibodiescan be interpreted
as an attempt of the organism to defend itself anew against the an,tigen.
The antigen, this time fixed through these antibodies in a new and more
noxious formation, will once again incito the nonspecific defense mechanism. First there will be the antiheterogeneousresponsewith its diphasic
phenomenonand, if once more this is not effective,a prolonged new lipidic
intervention will follow. If the quantity of heterogeneousformations is
gteat, the first phase of the diphasic phenomenoncan be so severe as to
cause death in a few minutes. If less severe,this first phase is followed by
the second,with chills and high temperature,As in all the antiheterogeneous reactions, the organism tries to combat the presenceof the noxious
factor-in this case, the flocculate produced by the antigen-antibodybond
-attempting to digest it through hydrolytic enzymes or to neutralize it
through constituents brought in during the second phase of the diphasic
180
RESEARcH rN pHysropATHoLoGy
DEPENSE
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pHystopATHoLocy
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DEFENSE
185
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REsEARcH rN pHysropATHoLocy
DEFENSE
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188 /
nESEARcHrN pHysropATHoLocy
DEPENSE
189
sams 1319as they appetr, as defenseprocessesin the blood can do, severe
local manifestations result. This may lead to necrosis and even rejection cif
alteredcells or tissues.These representthe very important difterenceswhich
exist between allergic processeswhich occur in the blood and those which
occur at the difterent levels following the appearanceof allergic antibodies
while the antigen is still present.
The fact that there will be no reaction when small amounts of the
allergic complex are progressivelyformed, as in casesin which antigensare
presentin the blood at the moment of appearanceof the allergic antibodi$,
is confirmed indirectly by the possibitity of prevonting severe systemic
manifestationsthrough skeptophylacticor desensitizationprocedures.The
introduction of very small amounts of antigen thus produces only small
amounts of complex at any one time, avoiding clinical manifestations.With
progressivedoses however, the antigens will fix circulating antibodies in
srrfficient proportion to prevent the formation of important amounts of the
same complexes after further administration of the antigen. The presence
of the secondphase of the diphasic phenomenon,with the exaggerationof
constituentsantagonisticto those presentin the first phase,will also act to
Prevent the occurrenceof an intensivefirst phasewhen the antigen appears
anew.
The situation changesentirely when antibodiesappear and the antigen
no longer is present. They can then accumulate in the blood in large
amounts. Thereafter, sudden appearanceof the antigen in sufficient quantity will form a large amount of the allergic complex and the subsequent
reaction can be so violent as to kill the subject.This occurs in anaphylactic
shock. When the antigenis limited to other levels,important local changes
can be induced.
The neutralizing immune antibodies, if manifestations already exist,
will prevent new ones from appearingand this will permit healing prooesses
to take place without further interference. The antibodies will prevent
manifsstationsat the respectivelevel if the antigen appearsagain.
Affinity ol Antigens
In the defense procsses,another factor intervenesto produce difierencesbetween responsesat difterent levels-the special affinity of antigens
for various cells, tissuesor organs.This affinity will determine not only the
levelbut also the individual entitieswhere manifestationswill occur. It has
to be emphasizedthat the independenceof the levelsor of groups of entities
in an organism goes so far as to allow the defenseprocessesto progressto
difierent stages.While defenseprocessesat the tissular level, for instance,
*,rffi
iffi kffiiiliffi *1h{LX##1{*"ir$itiil
190
REsEARcH rN pHysropATHoLocy
cannot go beyond the stage of prolonged lipidic response,those at the organic or systemic level can arrive at the allergic stage.we will see below
the importance of this unequal responseof the difterent levels.
The unequal capacity of different tissuesto manufacture allergic antibodies could be postulatedto explain the propensity for local allergic conditions. The ectodermic system appears especially inclined to allergic
responses,as seen for the skin. We tried to relate this to the natural richnessof these organs in sterols. This would explain the fact that the brain,
which is richest in sterols,seemsto show the earliest allergic manifestations,
which could be interpreted as resultingfrom early or more constant appearance of coagulantantibodies.
Besidesthese difrerencesin the responsesof various entities, an important factor intervenes in the induction of localized allergic manifestations. It corresponds to unequal affinity of the antigen itself for various
entities.This would localize the antigen in cells, tissuesor organs so that
when coagulatedantibodiesdo appear, the noxious allergic complex will
be formed locally in the sameentities.It seemsthat this localizationof the
antigen,such as upon nerves,kidney,lung, etc., is more important than the
capacity to produce antibodies in determining predilection of pathological
pr@essesfor specificcells, tissuesor organs.
one of the most interestingaspectsof the defensemechanismis the
relationshipbetween successivesteps.we could show, generally,that an
intensiveresponsein one step representsa favorable condition for appearanceof an intensiveresponsein the next step.It is a known fact that manufacture of immune antibodies is influenced by an inflammatory process.
This is the reason for the customaryinjection of tapioca, for instance,in
horsesduring their immunization for the production of therapeuticsera.
We could show that injectionsof lipids, lipid acids or insaponifiablefraction of placenta,or of organsof animalsof the same speciesfor instance,
manifestly hasten the appearanceof the next step in the defense against
the microbe.
It seemsclear that under the influenceof the lipids used,the agglutinins
appear in blood earlier and their amount increasesmore rapidly than in
the control animals.
Antigenic Factors
The interventionof different mechanismsin the defensehas led to the
suppositionthat each one would be induced by relatively specificfactors
presenteitherin the antigenitselfor appearingduring the defenseprocesses.
DEFENSE
l9l
':t.1fl;+ki#F*:..J*ir,#J,.,S*
; :'
192 /
n E s E A R c Hr N p H y s r o p A T H o L o c y
antigenicfactors. we will describehere very briefly severalsuch applications which were interestingalso becauseof the practicalresultsobtained.
Hydrolysis Products
we tried thus to utilize the productsresultingfrom the breakingdown
of body constituentsor of other materialsin order to induce through their
administration,the appearanccof the seconddefensemechanism.Applying
the dualistic concept, we separatedthus in the products of hydrolysis of
different materials,those with an acid characterfrom the group with basic
and alcoholic characters.Various materials were thus hydrolyzed using
KoH, NaoH or ammonia.The solublepart, separated,
was treatedwith an
acid and a precipitateobtained.After washingit, this precipitatewas red.issolvedby a.lkalizing
to a pH still below neutrality.This has represented
the
"acid fraction." Besidesacid lipids, this fraction containsalso acid protein
groups and even humic acids.
The part which remainedinsolubleafter treatmentwith KoH (separated from the soluble part) was treated with an acid. The part which
becamesolublewas then separated,
reprecipitated
by alkali, and partially
redissolvedby bringingthe pH, throughacidification,near 7. This representsthe "alkaline fraction." With differentdegreesof chemicalhydrolysis,
various fractions-more or less broken dsu/n-sps obtained for both the
acid and alkaline fractions.The degreeof this "digestion"has appeared
highlyimportant.The amountof the productsobtaineddecreases
for an insufficienthydrolysisas well as for a too highly pushedhydrolysis.
Accordingto the mechanisnrmentionedaboveit was expectedthat these
fractions,resultingfrom the breakingdown of body constituentsor of the
antigen and correspondingto the effect of the first enzymatic defense
mechanism,
would inducethe secondstepof the defensemechanism.This
would correspondin part to the interventionof the properdin systemand
of the lipidic defense.It has as characteristic
the fact that it would appear
only within a certaintime. The followingexperimentillustratesthis clearly.
The "acid fractions" of human blood, hydrolyzedby KoH was obtained
and then injectedintraperitoneallyto mice. At different intervalsfollowing
this injectionthe mice were inoculatedwith 3,000,000microbesof a fresh
c u l t u r eo f B a c .p r o t e u sI.n c o n t r o l st h i s i n o c u l a t i o n
w o u l d r e s u l ti n a I O O V o
lethalinfection.No protectionwas seento appearin the l6 hoursfollowing
the injectionof the "acid fraction."At thc l6th hour, J,2,,were protected.
This protectionincreasedwith time to be completeafter 22 hours, when all
the animals survived. This protection was still present after a few days.
The inoculationof thc nontreatedblood in the same proportion was
DEFENSE
le3
seenincapableof conferringthe samedegreeof defense,a fact which indicatesthe importanceof the breakingdown processin this ,.24 hours" defense.These resultsare similar to those obtainedby I. A. parfentjev with
malucidin,a product of hydrolysisof yeast.
Another applicationof the sameconcept was in the use of the lipidoproteiccomplexes.
In a group of researchstudies,we utirizedthe products resultingfrom
the bond betweenan antigenand a lipid, with the intent to obtain a lipidoproteic antigen and through it. a lipido-proteic defense response.Often
the mixture of the antigenwith the lipidic preparationsappearedsufficient.
Fatty acids, such as oleic, linoleic, arachidonicor eleostearic,acting
directly upon the killed typhoid microbeswere usually seento enhancethe
production of agglutinins and of specific immune antibodies. The same
effect was produced by lipoacids of the same spciesas the test animal.
Lipoacids of guinea pigs were especiallyactive in promoting the apparance
of agglutinins but less potent in inducing the appearanceof immune antibodies. The lipoacid fraction of bacteria such as B. subtilis, coli, diphtheria, acting in vitro upon typhoid killed microbes,led to the appearance
of antibodies against typhoid microbes but produced almoet no antibod.ies
againstthosemicrobesfrom which the fatty acidswere obtained.The lipo
acid fraction of tuberclebacitli bound to killed typhoid microbeswas seen
to induce agglutininsbut seemedto roduce and even prevent the appearance of immune antibodies.The same influencewas seen with the lipids
obtained frorn tlre seedsof Bixa orellana but was less accentuatedfor the
lipids from fish and squid. Butanol and especiallyheptanol were seen to
retard the appearanceof all antibodies,allergic and immune.
Allergic Precipitates
The injection of killed typhoid microbes agglutinated by a specific
serum was followed by rapid production of immune antisera. The serum
of rabbits injected with these mixtures prevents a lethal condition induced
in mice by intraperitonealiniectionof living microbesin much smallerdoses
than serum obtained with untreated microbes.
on the other hand, the injection of the same killed ryphoid microbes,
mixed together with a flocculate obtained, for instance, from egg protein,
and an antiegg precipitant-guinea pig serum-produces a much lcss
rapid appearance of antityphoid immune antibodies than injection of
microbes alone.
Another form of lipido-proteiccomplex, utilized as agent with the aim
to induce not a lipido-proteic responsebut a higher one in the defense
194
nEsEARcH tN
pHyslopATHoLooy
DEFENSE
r95
INFECTIOUSDISEASES
Toxic and Allergic Conditions
In infectiousdiseasethe antigenis a micro-organismwhich may bc a virus,
microbe, protozoa,mycct, etc., or even a product elaboratedby a microorganism.Thc responscof an organism to the presenceof an infectious
antigentendsto follow the same successive
stagespreviouslyoutlincd. If
the meansat the immediatedisposalof the organismare qualitativclyand
quantitativelysufficientto neutralizethe antigen,the entire processwi.ll be
resolvedasymptomatically.
othcrwise, the first stage of the defensercaction, the primary toxic diphasicphcnomenon.will bc set into motion.
According to the qualitativeeffectiveness
o[ this response,manifestations
will vary from simplesubclinicerJ
changesto clinicalreactions.If thc second
phaseof the diphasicresponsecannottake place.a prolongedform of the
first phasewill result.It correspondsto shock,which is encounteredonly
in very severeinfections.The rapidly lethalconditionresuttingfrom transfusion of massivelyinfectcdblood is an examplc.
The secondphasebringschill and fever.If thc secondphaseresponse
is qualitativelyinsufficicnt,the prolongedform ensucs,bringing fever,the
usualmanifestationof many infcctiousdiscases.
Thc fever persistsas long
as the nonneutralizedantigcnis present.In this stageof the defcnsereaction againsta micro-organism
or its toxins,the symptoms,althoughresultingfrom thc responscof the organism,are still directly rclated to the
presenceof the antigen in sufficientquantity.The quantity necessaryto
induccthe clinicalmanifestations
can be reachcdwithin a short time after
the penetrationof the antigeninto the organism.The toxic reactionthus
can appear in a few hours. Consequently,
there is no specificobligatory
incubationtime. The manifestations
will disappcarwhen the amount of
antigenis decreased
sufficiently.For somemicrobes,antibioticshave such
action,resuitingin a decreasein the amount of the antigenprcsent,and
consequently
in the disappearance
of the clinicalmanifestations.
A similar
decreasein the amount of the free antigenprcsentcan be obtainedby its
neutralization
throughspecificimmunescra,if available.Conseqently,
such
serahave curativeeffectsin infectiousdiseases
characterized
by a primary
toxic pathogenesis.
Allcrgic antibodieswill appearafter an obligatoryincubationperiod of
6 or more days. If the antigenis still prcsent.it may be destroycdby the
new defensiveantiheterogeneous
responses
mobilizedagainstthe resulting
allergiccomplex.ln this case,the appearance
of the allcrgicantibodiesre-
196
REsEARcH rN pHysropATHoLocy
sttlts in a kind of clinical crisis which can lead to the cessationof the disease. However, if this eftect does not occur, the appearanceof allergic
antibodieswill cause an increasein symptornsor in their gravity.
In cases asymptomatic prior to the appearanceof the allergic antibodiesbecauseof low direct toxicity or insufficient quantity of the antigen,
the diseasewill become cl.inically apparent only with the appearanceof the
allergic manifestations.The clinical condition thus will have an obligatory
incubation of 6 or more days, since this representsthe time necessaryfor
the coagulant antibodies to be produced. Since the manifestationsin such
casesare due to the allergic complex and not to the direct action of antigen,
they will be nonexistentor minimal during the incubation time. Due to the
allergic complex, the condition will not respond to specific immune sera
able to neutralize the antigen but ineffective against the allergic complex.
Specific immune sera are not curative for these infectious conditions of
alleryic pathogenesis.As already noted, only when administered before
allergic antibodies have appeared,during their incubation period, do these
sera have a marked preventive effect.
Thus the pathogenesisof an infectious diseasecan be toxic or allergic
in nature. The two pathogenicmechanismscan be identified easily tfuough
incubation time of major clinical manifestations. An infectious disease
which appears shortly after the entrance of the antigen without an obligatory incubation has to be corcidered, according to our concept, to be ol
toxic pathogenesiswhile one which appears alter an incubation time obligatory greater than 5 or 6 days has to be considered allergic.
Applying this concept, we have separatedthe clinical infectious diseases
into two groups, toxic and allergic, using incubation time as the criterion.
We wish to note here the great similarity in the incubation time for the
diseasesin each group. Most of the allergic group have an obligatory incubation time ranging from 6 to 14 days, which coincides with the usual
time neededfor the appearanceof the allergic antibodies.The incubation
time is independent of the fundamental nature of the etiologioal agentvirus, microbe, protozoa, etc.---or of the nature of their products+xo.
toxins, endotoxins, etc. This indicates that the principal factor in the
incubation time is the allergic pathogenic mechanism itself.
Based upon the criterion of obligatory incubation time, the following
diseaseswith brief incubation time have been consideredas having a toxic
pathogenic mechanism: diphtheria, botulism, anthrax (Bac. anthracis),
meningococcalinfections, cholera, some streptococcalinfections, dysentery
(especiallyShiga Kruse bac.), plague, scarletfever, pneumonia,etc. In the
allergic group, with an obligatory incubation time above 6 days, we find:
DTFENSE
l9'7
Xll
lNrrctrous Dtselsus
Incubation
Low
Dyphtheria
Anthrax
Botulism
GaseousCangrene
Plague
Erysipelas
Dysentery
MeningococcicInf.
Cholera
Pneumococcus
Inf.
Ohligatoryubove 6 duys
Typhoid
-f
etanus
Pcrtussis
Glander
Tularenria
Leprosis
Typhus
Rabies
i\'teasles
Munrps
Poliomyelitis
Smallpox
Chickenpox
Recurrentfever
198
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If the paralysis were induced by direct action of a thermoatabilefraction of the toxin, then higher doses of the fraction should reduce, or at
least, not increase the incubation time. This paradoxical fact can be explained simply through the mechanismof allergic pathogenesis.It is a fact
common to immunological reactionsthat an organism has gteater difficulty
in manufac{uring any antibody when very large amounts of antigen are
Presentthan when smaller amountsare involved. This difficulty is translated
into a longer time necessaryfor the appearanceof the antibodies.As seen
in our experiments,in the caseof an a.llergicreaction, this difficulty in the
manufacture of coagulant antibodies would result in a longer incubation
time.
The localization of the auergic manifestation as paralysis can be explained in part through the affnity of toxin as antigen for nerves and in
part through the participation of the nerves in the allergic reaction. The
levels at which the diphtheria toxin acts seem to be tissular, organic and
systemic,with preferencefor the adrenals,inducing characteristicsuprarenalitis. When coagulant antibodies appear, no manifest systemic allergic
reaction will occur with the antigen still present in the blood. The allergy
will be manifest,however,at the lower tissuelevel and especiallyin the
nearby nerves. Antigen must be presentin the nerve at the moment of
appearanceof the coagulant antibodies if paralysis is to occur. This can
be demonstratedby using sensitizingand triggeringinjectionsof toxin in
animals.
we sensitizedguinea pigs to heated and unheatedtoxin by injecting
relatively small amounts intravenously.On the sixth or seventhday, another small quantity of the same toxin was injected, this time near the
sciatic nerve. The total amount of toxin was far below the lethal dose. Two
or three days later, paralysisdevelopedin the injeaed timb in a high proportion of animals while no such paralysiscould be observedin animals
injected only intravenouslyor with the same total amount of toxin at once
in the limb. In other experiments,the daily injection of small amounts of
toxin, whetherheatedor nonheated,near the sciaticnerve, induced paralysis although the total quantity of toxin was much lower than that which
ordinarily would induce para.lysis
in any similar animal. paralysisappeared
in thesecasesafter an incubation period of about 12 days. The animals
sensitizedby one or more injectionsof heatedtoxin respondedto the nonheated as triggering injections and vice versa, indicating that antigenic
propertieswere responsiblcfor rhe paralytic allergic manifestation.
In humans,anti-diphtheriaserum,eftectiveagainsttoxic manifestations.
had no effect upon paralysisonce it had appearedbut is very effective in
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DEFENSE
201
202 /
R E S E A R c Hr N p H y s l o p A T H o L o c y
8th day, the same serum prevented the appearanceof the allergic form of
the experimental disease.
Tetanus
In the light of our concept, we studied tetanus pathogenesisin an effort
to explain the classicallyemphasizedseparationbetweenthe so<alled small
and big animal disease.(39) In mice, tetanushas a short incubation period
and is manifestedby localized contractions,while the diseaseof so-called
"big animals" starts with trismus after an incubation period of more than
6 days. Based upon incubation times, we consideredtetanus to be the toxic
form in small animals, the allergic form in large animals.
we could, in fact, induce a condition in mice manifested by trismus
and epistotonus,and having an incubation period longer than 8 days, by
daily repeatedintravenousinjectionsof small amountsof toxin. The special
affinity of this toxin for the nervous system,and the strong bond between
nervous tissue and toxin, limited the response to the antitetanic scrum
even of the primary form. If injected in time, however, the antitetanic
serum controlled this primary form. The same serum appearsto be highly
effective in the prevention of the condition in animals which have been
prepared for the allergic form, provided it is administered before the allergic manifestations have appeared. The same senrm is totally inaaive
once the allergic condition is present.
Rabies
Under all circumstances,rabies needsan incubation time of more than
5 days. From our point of view, therefore, it has to be considered an allergic condition. when the rabies virus was passedrepeatedly through the
brains of rabbits, incubation time became continuously shorter and ultimately was fixed at 6 or even 5 days. Classically,this progressivelyshortened incubationtime is interpretedas being due to progressivelyincreased
virulence of the virus after these passages.In the tight of our concept of
the pathogenesis
of infectiousdisease,a reductionof incubationtime is the
result of increasedvirulence onJy in casesof primary direct toxic pathogenesis.In the allergiccondition,which has an entirely differentpathogenic
mechanismwith the incubation period related to the time necessaryfor
the body to producecoagulantantibodies,a shorterincubationwould correspond to a different change.It resultsfrom a greaterfacility of the organism
for manufacturingantibodiesagainstthe infectious agent. In the case of
rabies, a short incubation period of 5 days for a "fixed virus" would mean
that the organism is able to manufacture allergic antibodies more easily,
DEFENSE
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enzymes,lipids, allergic antibodiesor even immune antibodies-a graduated seriesof changesin an organism'sown entities can be induced. of
the factors which intervene in the heterogenizationof such entities, we
have studiedthe lipids in particular. An antigenicrole for lipoacidscould
be shown in many experiments.For example,suspensionsof red cells or
cellsof different tissuesof guineapigs in a concentrationof I gram of cells
to l0 cc. of saline were prepared,At the same time lipoacid preparations
wereobtainedin the followingmanner.5 cc. of a Z% solutionin alcoholof
difterentlipoacidsor mixturesof lipoacidswere added to ll0 cc. of water
and the preparationboiled under low pressureuntil reducedto 100 cc.
The cell suspensions
were administeredto guineapigs in four injections
at weekly intervalswith no seriousmanifestations.
The same was done for
the lipoacidsabove. A preparationwas obtainedthrough the action of the
lipoacidsupon the cell suspensionin the following manner. 5 cc. of the
colloidallipoacid aqueoussuspension
were addedto 5 cc. of the suspension
of cells of different tissues.To 5 cc. of red cells, only I cc. o[ the lipidic
suspensionwas added. The mixture in each case was incubated for two
hours at 37"C and centrifuged.The cellular residues,separatedfrom the
supernatantfluid, were resuspendedin saline zr.ndkept frozen. While one
injection of the so-treatedcells showed no noxious manifestations.consecutiveinjectionsat weekly intervalswere seen to induce, in less than a
month, manifest changesin the respectiveorgans. with the red cells, a
marked anemia was induced. oleic and linoleic acids, and the lipoacids
from human placenta,cow liver or total body of guinea pigs had only a
slight effect in inducing organ lesions.The lipoacids obtained from Bixa
orellana and especiallyfrom the tuberclebacilli led to seriousdamage in
the respectiveorgans and resultedin death usually in less than 3 weeks.
Tuberculin in thesecaseshad the same effect as lipids obtained from tuberclebacilli.
The degreeof heterogeneityof the lipoacid appearsto be the factor
which determinesthe stageof defenseinduced,The lipoacid-cellcomplex
acts as an antigen.with the type of cell determiningthe organ where the
abnormal changesoccurs, and the lipid determiningthe character of the
occurringreaction.Dependingupon the lipoacid. the effect will vary from
minimal tissularlesionsall the way to massivedegenerative
changesleading
to death.
The intervention of a bond betweencells and lipids apparsevident
when acid lipid preparationsare injected repeatedlyat week.lyintervalsin
the sameorgan. Lesionsare obtainedwhich are similar to but lessintensive
than thoseproducedby the cell-lipoacidcomplexes.
DEFENSE
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R E s E A R c Hr N P H Y s T o P A T H o L o c Y
DEFENSE
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RESEARCH IN
PHYSIOPATHOLOGY
o
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Flc 7E. Wound chloride ctrrve. The curve of the amount of the chlorides present
in skin wounds in rats correspondsto the average value obtained in 6 rats for each
figure. A first phase, with high values correspondsto tbc offbalance D. This is followed by a second phase characterizedby an offbalanceA. A variation in the curve
correspondingto the 8th day is constantly seen as corresponding to an allergic reaction. The values representmgs. of chlorine per 100 gr. of weight of the wet malerial.
PROBLEMS IN CANCER
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xESEARcHtN PHYsropATHoLooy
Grafts in Humans
The grafting of cancerouscells in normal humans usually leads to the
appearanceof a growing tumor which (315) after a period of days, almost always sufters the same fate as a moderately heterogeneoustransplant: death followed by resorption or expulsion. The time when this processo@urs indicatesthe allergic natureof the defensemechanism.It is highly
probable that, without the intervention of this efficient allergic defense,the
cancerousprocess would have continued to evolve.
Such continued growth occurs if grafts are made in subjects already
having their own cancerousprocess.This would indicate that the allergic
defensemechanism against the graft is no longer operating in these cases.
The fact that a canceroussubject acceptsa new tumor graft while the
normal one rejects it indicates that defenseprocessesare different for the
normal and this canceroussubject.The inability of thesesubjectsto reject
a grafted tumor through an alleryic responseappears to be the major immunological difference between the normal and the cancerous subject. Still
more important is the fact that in cancer patients,the anomaly would correspond to a lossof the capacity to reject the grafted tumor which the normal
subjectseemsto have.
In trying to determine the nature of the immunological anomaly in the
evolution of a spontaneouscancr in patients, we have to relate it to this
loss of the defenseprocessesas seen above.
In studying this occurrencein general,a loss of defenseagainst:rn antigen can be conceived to occur for any of the three different mechanisms
involved in defense:primary, allergic or protective.In caseswhen this takes
place, the loss of the protective stagewill take place first. The allergic defense will be affected next and finally, the primary response.This explains
why the inability of an individual to achieveone stageof the defenseleaves
the defenseresting in the immediately previous stage.The inability to manufacture protective globulinic antibodies,for instance, will leave an individual in the allergic stagewhich, in the developmentof defense,preccdes
manufactureof the immune antibodies.This results in a potential allergic
condition if the antigen is absent or an actual allergic condition if the antigen is present.We have seenthat this occurs in most of the chronic infectious conditions.Similarly,with the inability of an organismto manufacture
coagulantantibodies,the defenseremains in the previous defensestage,the
primary lipidic one.
Before going further, we have to discussa factor believed by many
authon to be involved also in the defensemechanismagainstcancer. A few
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DEFENSE
215
216
nESEARcHrN pHyslopATHoLocy
present,indicatinga prolongedprimary, toxic stage.The lack of local reaction seenat the site of the growing transplantin the canceroussubject at
the time when the normal individual kills or rejects the transplant points
to the fact that the anomaly residesin qualitative inability to manufacture
allergicantibodies.
The next problem was to investigate the reason for the failure of
allergic defense against the tumors. we could show that the cancerous subject has not lost the capacity in general to manufacture coagulant antibodies.Even subjectswith very widely spread cancer were able
to respondwith a local skin allergicreactionto a secondinjection of an antigen (proteins from mollusks) made more than ten days after a first preparatory one. (Note 7) Their inability to fight transplantedcancer cells
through a similar allergic reaction indicatesthat the loss of this capacity
is not generalbut relatively specifictoward the cancerouscells. The lack
of an intensive inflammatory process, as well as the existence of high
amountsof lipids in the canceroustissue,also would indicateindirectly an
inability of the canceroussubjectto resolvethe existing immunological
problem of fighting cancer through an allergic reaction. The presenceof
large amountsof lipids indicatesthat thc defensemechanismhas been
arrested in the stage of pronounced lipidic predominance. Abnormal
amountsof lipids thus could representan indirect meansof recognizingthe
failure of an allergic responseto cancerousentities.
The next problem was to try to determinewhere in the organization
the failure occurs.The different levelsof the organizationare independent
to a certaindegreeand passageof an abnormalityfrom one level to another
induceshierarchicprogressionof the condition. This has posed the problem of the progressiveloss at the differentlevelsof the natural capacityto
defend againstcancer. Recently many investigatorshave shown that cancer cells passinto the lymphatic systemand into the generalcirculation in
a much higher proportion than had been suspectedbefore. Malignant cells
in the circulation are destroyed,however,by the defcnsemeanswhich are
not lost at this level.The samepatientthus may still have an activelygrowing cancerat the tissularlevel,indicatingthat this defenseprocess,although
successfulfor the higher levelsof the organization,does not interveneat
this lower level.
The hierarchicprogressionof cancercan be seenas a proefessiveloss
of the immunologicaldefensecapacity.while the organism conservesthe
capacityto fight at a higher level, a lower hierarchicentity no longer opposesthe cancerouscondition. It is not the absenceof cancerouscells in
blood or organs which explainsthe lack of an explosivespreadof the dis-
DEFENSE
217
218
RESEARcH rN pHysropATHoLocy
DEFENSE
219
virus will be prevented from reaching the cells. Once tbe circulating antibodies iue no longer present, the cells cease to be protected. For an efficient defenseagainst viruses an immunity within the cell appears thus to
be indispensable.The use of dead virus vaccine will induce only systemic
immunity, which can be recognizedthrough the circulating antibodies. It
is unlikely that the killed virus enters the cells. It does not aftect them,
and consequentlydoes not induce cellular immunity. Even a mild cellular
infection with living virus will give the necessarylongJasting cellular imrnunity. This would explain the need for living and not killed vaccinesfor
viral infections, as first postulatedby Pasteur.
A similar level immunity can explain the differencesseen between the
immunity resulting from the use of microbial vaccines and that produced
by natural disease.Typhoid infection gives lifelong immunity; the vaccination, only relative and temporary immunity. An explanation can be found
in the fact that, in the disease,along with the septicemia,manifestchanges
occur in organs and tissues.Spleenand lymphatic tissuesare highly affected
in typhoid and it is possible that the developmentof the defense at their
level would explain the lifelong immunity that follows the natural infection.
rn cancer, the problem would be to induce not a systemic defense,
which is still present for invasive cancer, but an effective tissue or even
cellular defense.Immunologicaltreatmentof cancer would have to make
tissular and possibly cellular levels regain their capacity to defend themselves through efficient allergic responses.The immunological prevention
of cancer would lie not in the creation of this defenseor in increasingit
quantitativelybut enhancingit qualitatively.A successfulallergic defensc
at this level apparently would have a preventive and even curative effect.
The use of lipids in the induction of the defensemechanismagainsttissues
has an interestingapplicationin cancer.A systemictreatment with lipids
or lipoids can change the defenseresponseso that it can be effective at
a specific level where it is otherwise inadequate.For invasive cancer, the
lipid activity must be induced at the cell level. The active lipoids for this
purpose are those with a high affinity for the cancerouscell.
As abnormal cells in general show similar capacity to bind the lipoids
administered,this general affinity becomesa handicap if abnormal entities
other than cancerouscells are present. These considerationshave led us
to attempt to use methodswhich will insure the activity of lipids at the ccll
level.
In one of these methods,the chosenlipoids are brought directly into
contact with cancerouscells through local injections into the tumors. SingJe
injections produce only limited changes in tumors. Local injections re-
22O /
REsEARcH rN pHysropATHoLocy
peatedso as to insure the presenceof the lipoids once, and then again 15
days later, are required to induce an eftective response.The lipoids or
lipids are so chosen that, when bound to body constituentsthey will induce allergic or immune defenseresponses.The acid lipids of tubercle
bacilli, bixine or guinea pigs are especiallyprone to induce allergic reactions,while the lipids of microbes-such as coli, typhoid or diphtheria
-produce immune responses.
In another method, lipids chosenwere bound to cancerousentities in
vitro. Cancerouscells were obtainedand treatedin vitro with lipids under
whoseinfluencethe body is able to manufactureallergic or immune antibodies. colloidal suspensionsof the lipids or lipoids were prepared as
mentionedabove,mixed with suspensions
of cancerouscells, kept at 37.C
for a few hours, then separatedfrom the non-fixedlipids and injectedinto
patients. In order to obtain good results it was necessaryto inject this
material at least twice, at an interval longer than two weeks, in order to
insurean allergicreactionagainstthc cell-lipidpreparation.While a single
injection produced good resultsonly in a very small number of cases,repeated injections were manifestly more effective.When canceroustumor
cells could be obtained through biopsy from the patient, we used thenr
for the in vitro treatmentwith lipoacids.when biopsy material was not
available,we used cancerouscells of similar origin as the tumor of the
subject,preferablypooled.
The condition for successof these methods has appearedto be the
presenceof the cell-lipidcomplex at the moment of appearanceof antibodies. This is assuredonly by the repetition of the injection. Another
interestingaspectof the immunologicalproblem in cancer, related to loss
of the natural defensemechanism,is the loss by cancerentitiesof their
capacity to utilize certain elements known to intervene in the defense
mechanism.The role of magnesiumin the properdin system, copper in
cytochromeoxidase,of calcium in generaldefense,suggestsa correlation
betweentheir deficientutilizationin cancerand the loss of the defense.We
will discussthis problembelow,after reviewingthe pharmacological
aspect
of theseelements.
CHAPTER
T H E C O R R E L A T I O Nt s E T W E E N
T H E B A S I CC O N C E P T S
rTrl
rs A cLEARinterrelationship
l[
betweenthe four basicconceptspre"r^u
viouslydiscussed
which permitsus to considerthem togetherand to establish a unified viewpoint.For all four can be scento representdifferent parts
of the same fundamentalproblem in biology: the manner in which an
entity resolvesenergeticdifferencesbetweenitself and the environment.
we have seen that, in the framework of fundamentallaws governing
nature, matter can be consideredto correspondto isletsof heterotropyopposingthe homotropictrend of evolution.Conservationof an existingentity
appe:us to be the principal meansby which heterotropycan be achieved.
And heterotropyis fulfilled, specifically,through maintenanceof the constantsof entitiesas valuesdifferentfrom thoseof the environment.
H i erarchic Organization
The continuoustendencyof natureto progresstoward maximum homotropy has made the conservationof existing entities a persistentlyacute
problem. The problem posed by the progressivelychanging environment
cannot be solvedthroughchangeswithin entitiesthemselves.
Any "adaptation" of the entity itselfwould affectits constantsand, consequently,
would
be contrary to the fundamentalpurposeof heterotropy.Nature has resolved
the problemin an entirelydifferentway. Sincethe entity itself must remain
unchanged,and yet the influenceof the environmentmust be conteracted,
nature has made use of hierarchic organization.Secondaryparts, reproducing the immediateenvironment,are joined to existingentities.often
surroundingthem and acting as buffersagainstenvironmentalinfluences.
Through theseadded secondaryparts,hierarchicentitiesare organizedso
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T H E C O R R P L A T I O NB E T W E E N T H E B A S I C C O N C E P T S
223
cntity, which is part of the complex organism,suggeststhat theseconstituents entered into the formation of these entities as the result of thcir
interventionin the defensemechanism.Thus, it can be conceivedthat. in
its phylogeneticdevelopment,each entity has passedthrough a succession
of defense phases in which specific groups of constituents-nzymatic,
lipidic, lipido-proteicand proteic-have been predominant.In actual organization,while all higher entitiescontain fundamentallythe sameconstituents, different substancesare predominant at different levels. This can
be explainedby the predominanceof a particular defensemechanismat a
particularlevel.According to this view, this defenseis principally in the first
stage,that is, of enzymaticnature, for most endodermicformations. It is
in the prolongedlipidic stagefor ectodermicformations,lipido-proteicfor
the reticuloendothelialsystem,and proteinic for cells. Through thesecorrelations,constituentscan be more completelyintegratedin defcnse.
The kind of special defensedevelopedfor the different levels of organization, through predominant specific constituents, has not been
followed by a total discard of the other constituents,which do not have
such roles. Instead,the latter have been retainedin the entities in smaller
amounts and in inactive forms. This confers upon the entity the capacity
to mobilize these constituentsand use them when the need to respond
to an acute emergencyarises.Pre-fermentsand cven ferments in mitochondria; fatty acids and anti-fatty acids bound as esters;lipido-proteins
and proteins in various combinations-all these are inactive constituents
which can be changed easily into active agents. when fighting a new
noxious intervention,an entity will resort to liberating or activating these
constituentskept in reserve.Each entity and level of organizationdoes
this indepcndentlyof other entitiesand levels,yet constituentsactivatedat
one level can act at other levels, too. The successor failure of defense
especiallyin its first stages,dependsnot only on the intrinsic value of thc
constituentsavailable, but also on the capacity of the affiicted entity to
utilize these means by activating them. Although activation processesbecome strikingly evident in abnormal conditions similar processessecm to
be important even in the maintenanceof existingentities.
Dualism, as we have seen, characterizesboth normal and abnormal
physiology.That which is considered"normal" is the result of an altcrnating intervention of two groups of opposite constituents,producing an
oscillatory movement and a dynamic balance. The dualism seen in abnormalities,when one or the other opposedfactor is persistentlypredominant, is related to hierarchicorganizationand the defensemechanism.
Dualism results from the intervention of two fundamental forces in
224
nESEARcHrN pHystopArHoLoGy
CHAPTER
S H O ( ]K
r
ll* ro,r, oF THE pRocRESs
rcalizedin the lust decade,shock remains
one of the most challengingproblemsin medicine.That lipids have a
critical role in shock pathogenesis
seemsclear from a long-term study
which beganwith an investigation
of the activity of fatty acids in the induction of the abnormallydark color of blood secn in shock.The results
of this study will be discussedhcre not only becauseof the intrinsicinterest of the problem of shock itself but a.lsobecauseshock often represents
the terminal phaseof cancer as it does of many other diseases.In this
presentation,we will try to remain as much as possiblewithin the framework of our direct contributionto an understanding
of shock. A portion
o f t h e s er e s e a r c h ewsa s p u b l i s h e di n 1 9 4 3 .( 4 0 )
In studyingthe very complex phenomenonof shock, one has to consider a seriesof well-definedproblems.Shock has been related not only
to a large number of causesbut also to a seriesof vcry varied clinical
manifestations.
An initial problem was to determinewhetherthere is any
common relationshipbetweenthe different types-between the shock, for
instance,which kills a subject within a few minutes after a severesudden
trauma, and the shock that kills in days through profound systemicmetabolic impairments.What is common to, and what is different between
them, from the point of view of pathogenesis?
What constituentsintervene
and how, in shock?Theseand many other problemshave beenapproached
systematically.
T-vpesol Shock-As a starting point, we attenrptedto classify the
types of shockand found an interestingrelationshipaccordingto the time
that is, the intervalbetweenapplicationof thc noxious
of their appearance,
Three typescould be identifiedwith
stimulusand onsetof manifestations.
this criterion.
225
226 /
R E s E A R c Hr N p H y s r o p A T H o L o G y
sHocK
227
conditions.
The next problem was: could a common pathogenicmechanismbe
recognizeddespite the greatly varied manifestationsof these three forms
of shock?
Shock Mechanism
We saw one primary correlation between the three clinical types of
shock in the fact that sometimesone type is followed by another. Superacute shock, if not lethal, may be followed by acute shock which, in turn,
can change into a state of shock.
But it was the chemical analysisof blood, organs and entire bodies of
animals killed by any of the three types of shock which indicated the possibility of a mechanism common to all three. A low antitryptic power of
the blood, and the presenceof substancesresulting from protein hydrolysis
were found to characterlznall 3 types of shock. Additionally, ut increase
in the amount of free fatty acids, and the presenceof abnormal members,
occurred h all three types.
Fatty acids were studied from the point of view of the reciprocal position of their double bonds, through the oxidative fission method mentioned
previously. The appearanceof oxalic acid following oxidative fission indicates the presence of conjugated double bonds. The oxalic acid index
obtained indicates the proportion of these conjugated double bonds. In
normal rats, this oxalic acid index usually is zero in the total amount of
fatty acids; in normal mice, values below I are seen. In all animals in
shock, even in cases of superacuteshock followed by sudden death, the
oxalic acid index is invariably much higher. Furthermore, the death of an
animal in acute shock or state of shock appearsto be related to the presence of a critical oxalic acid index, indicating a concentration of abnormal
fatty acids incompatible with life. Whether it appears in a relatively short
time as in acute shock, or many days after the noxious intervention as in
the state of shock,the oxalic acid index found in dying animals is between
14 and 17. Such high values are not found in superacuteshock but the
oxalic acid still is markedly increased.Thus, the presence of hydrolytic
processestogether with abnormal fatty acids appears to be a common
pathogenic factor for the difterent forms of shock.
Pathological Clanges
The three types of shock-because of the presencein all of hydrolytic
processesand abnormal fatty acids-<ould be related to the first phase of
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RESEARcH rN pHysropATHoLocy
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231
andof sodium ions into the intestinesa@ountsfor the physiological"alkalinetide" associatedwith digestion.
when chloride ions are pathologicallyfixed to abnormal fatty acids in
theblood, they can no longer be dissociatedand secretedby the stomach
in the form of hydrochloric acid. Instead,they remain bound to the fatty
acidsand accumulatein this form within the gastricmucosa.The multiple
rnilliar gastric mucosal ulcerations in the state of shock results from the
interventionof these abnormal fatty acids brought into the mucous membraneby the chloride ions to which they are bound. The ulcerationsare
causedby the catabolicaction of fatty acids.Thus, the first phaseof abnormal sodium chloride metabolismleadsto the characteristicmultiple gastric
ulcerations.
The secondphaseis relatedto the metabolismof sodium. The sodium
ions are secretedas carbonatesby the pancreasand intestinal mucosa in
the fint part of the small intestine.In the state of shock, becausethey do
notencounterthe chloridesnormally coming from the stomach,they remain
as carbonates.As sodium carbonateis accumulatedin the first portion of
the small intestine,a local alkalosisoccurs, leading in turn to an important
loca.lretentionof water. It shouldbe noted that this is a very differentsituationfrom achlorhydriaor hypochlorhydria
in which,while the chlorideions
arenot secretedinto the stomach,no excesses
of sodium ions appearin the
blood or in the intestines,and consequentlyno local alkalosisor fluid
accumulationoccurs.
The differencebetweenthe systemicand tissueprocessesin shock lies
in the localizationof the abnormal sodium chloride metabolism.In tissue
anomaly,the separationof sodium chloride takes place between the cells
andthe pericellularstructures.At the systcmiclevcl, it occurs betweenthe
stomachand intestines,
with the blood servingas intermediary.This mechanismexplainsthe larger amountsof water which distendthe upper parts
of the intestine,as observedin autopsiesof animalswhich have died in this
form of shock.
The close similarity betweenthe abnormal processesthat take place in
sodiumchloride metabolismat the tissueand systemiclevelsprovidesthe
basisfor another working hypothesisconcerningthe mechanismin superacuteshock. We have seenthat the productionof vacuolesin cells characterizesthis latter form of shock. The unequal cellular permeability for
chloridesand sodium in their dissociatedform is known. Chloride ions can
circulatemuch more easily betweencells and the pericellularspacesthan
can sodium ions. An initial effect of the intervention of abnormal fatty
acidsin cellularpathologyis the fixationof chlorides.At the sametime, an
232
RESEARcHrN pHysropATHoLocy
sHocK
233
same pathogenicprocess,abnormal sodium chloride and consequentabnormal water metabolism, produces such different manifestations in the
various types of shock. It must not bc forgotten howcver, that in the last
analysis,the abnormalitiesin sodium chloride and water metabolismresult
from the interventionof abnormal fatty acids.Fatty acid intervention,together with the abnormal sodium chloride and water metabolism confirm
the unitary pathogenesisof the three forms of shock.
Other Changes
Other changesassociatedwith shock also can be related to the influence
exercisedby abnormal fatty acids. The appearanceof rouleaux of red cells
may be easily explained by fatty acid intervention. It is the replacementof
the nonpolarity normally present at the surface of the red cells by a dipolarity which results in the formation of the rouleaux. This can be induced
by fatty acids in vitro. Sludge formation would represent a still more advanced step in this same processand would apparto result from a polypolarity at the surfaceof the red cells.Sludgeformationshave been induced
in vitro by fatty acids added in larger amounts to plasma. (Note 2) They
contribute to the circulatory impairment considered to be an important
factor in the tissular respiratory troubles seen in shock.
We have noted that the richnessin free fatty acids interferes with the
ability of the red cells to keep oxygen fixed, a fact which would impair its
transport. This, together with hemoconcentrationand circulatory impairment, has been found to account for the black color of the blood in shock.
(Note 3/ The clinical manifestationsare characteristicof offbalanceD.
Experimentally Induced Shock
The hypothesisthat the three types of shock are caused by the intervention of the same factol-3lnospal fatty acids-has been further confirmed experimentally. The cellular changes that characterlze superacute
shock can be induced by the rapid introduction into the blood stream of
c-ven minislsl amounts of fatty acids in preparationsin which they are
bound to plasmaconstituents.
Pooled heparinizedplasma of mice was treated by stirring it in a nitrogen atmospherefor one hour with a preparation of conjugated trienic fatty
acids. The nonbound fatty acids were separatedthrough short centrifugation. The plasma was injected intravenouslyin mice. For control, plasma
treated under the sane conditions with stearicacid was used. While control animals did not show any apparent discomfort, the mice injected with
the plasma treated with conjugated fatty acids died immediately, in most
234
xEsEARcH rN PHYsToPATHoLooY
sHocK
235
O
C H A P T EI R
RADIATION
"ln
lL Hp sruoy
RADr^rtoN
237
g
o
v,
0
I
6
t
.6
L
fa
I rradiated
fbnirradiated
i l aY e
Length
(np)
Fto. 79. Irradialion ond conjugation in vilro. Spectral analysesin ultraviolet of samp l e s o f c o m m e r c i a l l y a v a i l a b l el i n o l e i c a c i d ( w i t h s m a l l a m o u n t s o f l i n o l e n i c a c i d
present) irradiated with gamma rays from 80 mgr of platinum filtered radium/10 cc,
f o r 6 d a y s a t r o o m t e m p e r a t u r eF. o r t h e a n a l y s e sd, i l u t i o n so f 0 . 0 0 2 9 bi n a l c o h o l ,w i t h
alcohol as reference, were used. The absorption spectra of the irradiated (-)
linoleic acid compared with the nonirradiated ( . . . . ) shows the appearanceof conjugated trienes recognizedthrough the characteristicpeaks.
238 /
n E s E A R c Hr N p H y s l o p A T H o L o c y
-l
,tl
RADIATION
I
,t-1
30
E
an
c
o
F
"-*
"l
-l
uo']
t
I
I
'ol
'ol
too
rl
I
I
I
I
I
I
I
I
I
I
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oo1
tn
t-'\
,
t
,
t
,
I
I
20
20)
.9
239
-aa
c
.9
t
a
t
t
t
320
300 ?30
240
220
200
ysis. When the same mixture of fatty acids was treated by the usual
chemical methods ernployedto produce conjugation,i.e,, with potassium
hydroxide in ethylene glycol or glycerol ( 4l ), the spectral analysis
showed that the preparation contained fatty acids having between 2 and 6
240
RESEARCH TN PHYSIOPATHOLOGY
T,rsle XIV
Fatty Acid
Type of
Radiation
Sourceof Radiation
Exposure
Time
(Days)
Oxalic
Acid
mg/E
Fatty
Acid
6
8
l5
4
0
8.3
9.9
13.3
9.85
7
27
4.3
6.8
53
12.4
5
8
0
5.2
8.0
Linoleicacid
tt
tt
tt
t
tt
r
tt
gamma
gamma
gamma
beta
alpha
x-ray
tt
x-ray
50 mg Radium
120 mg Radium
120 mg Radium
25 mg Radium
in Monel metal
150 uc Thorium X
5000r daily--deep
therapymachine
5000r daily--{eep
therapymachine
Cottonseedoil
t
tl
t
tt
gamma
gamma
80 mg Radium
80 mg Radium
Fatty Acid
Linoleic Acid
l0 cc.
l0 cc.
Exposure
time:
Days
Oxalic
Acid
mg g
Fatty
Acids
0
4
6
8
0
4.5
6.2
9.9
tt
15
20
r3.3
t4.4
1 5 0m g R a
,,
6
15
8.2
16.3
Sourceof
Radiation
0
1 2 0m g R a
RADIATION
241
IO
20
30
c
o
10
(a)
tn
.n
E
50
v,
E
.tt
t-
60
ia
70
I rradiated
Noninradlated
80
s
28o
260
i t a v e L e n g t h
2,to
(D!)
220
Frc. tl. Spectral analysis (0.002Vo in alcohol) of a mixture of. latty acids lrom cod
with 100 mg platinum
l i v e r o i l , u n t r e a t e d( . . . . ) ( a ) a n d i r r a d i a t e d( b ) ( - )
filtered radium/3 cc, at room temperature for 6 days. The analysis shows that the
conjugation which takcs place leads to the appearanceonly of conjugated trienes,
in spite of tbe presence of di-, tri-, tetra-, penta- and hexaenic unsaturated fatty
acids as shown by the absorption spectrum of the same mixture after chemical conj u g a t i o n w i t h p o t a s s i u mh y d r o x i d e i n e t h y l e n eg l y c o l . ( c ) a s s e e n i n F i g . 8 2 .
c) The changesinduced in fatty acids are essentiallythe same regardless of radioactive source. Thus the effect upon a linoleic acid preparation
containingsomelinolenicacid was the samewith alpha particlesof Thorium
X, beta rays from radium in monel metal, gamma radiation from platinum
242
xESEARcH IN
PHYSIoPATHoLocY
30
40
50
C
.9 -^
.3bu
c
o
Fzo
)
80
90
400
380
36U.
340
320
300
28c
WoveLength(mp)
260
240
220
243
RADIATION
l0
20
30
/r'\'.
f .,
I '
c
o
.a
40
att
/,'
l t
\'.\
\'..i
t v
t
t
,
ah
t
E
U'
)
,
l
bU
I
g
I
C'
I
t
I
I
i-
r-
60
t
}Q
m
I
l i
I r
80
l i
I t
t^'
Ir r a d i a t e d
llonirradiated
i
90
---.-'2
oo
2qo
U a v e l e n g t h ( m p )
Fro. 83. Absorption spectra (O.OO2Voin ethyl alcohol/ethyl alcohol) in ultraviolet
o f c o m m e r c i a l l ya v a i l a b l el i n o l e i c a c i d ( w i t h s m a l l a m o u n t o f l i n o l e n i c a c i d ) n o n with beta particles from 25 mg monel metal
irradiated ( . . . . ) and irradiated (-;
filtered radium/10 cc at 37'C for four bours. Some conjugation occurs in thc control
when kept in the incubator,
raiff$.r,4,{a{tfr
{iffi#(ffi'}*$itl's i#t1'
244
nESEARcH rN pHysropATHoLocy
beforeexposure.Groups of four treatedanimalswere sacrificedperiodically, startingimmediatelyafter irradiation,at 2, 6 and 24 hours aftcr irradiation,and eachday thereafteruntil all animalsdied or had beenkilled.
During this time, nontreatedcontrol animals,kept under the same
conditions,were also sacrificed.The quantity of conjugatedfatty acids
c
o
e
c
C'
L
>e
f a v e l e n g t h ( z r A )
F r c . E 4 . A b s o r p t i o n s p e c t r u m o f t h e s a m e m a t e r i a l a s i n F i g . E3, non-irradiated
( . . . . ) a n d i r r a d i a t c dw i t h o l p h a p a r t i c l e sf r o m 1 5 0 m c T h o r i u m X / 1 0 c c a t r o o m
t e m p e r a t u r ef o r 7 d a Y s . ( - )
RADr^rtoN
245
*
r
xt
X
r r r
l
,
x
8
r.ft
:
x
c
;
.!
x
o
I
E
(t
L
Ftc. E5. Futty acid conjugution indttted hy-irradiation in vir'<t.Changes in the oxalic index of total fatty acids of rats irradiated with a letlrcl dose (1500r). Sacrificed
at different intervals, the oxalic index of their fatty acid shows progressivelyincreasi n g v a l u e s ,T h e a n i m a l sd i e w h e n t h e i n d e x h a s a r r i v e d a t a c r i t i c a l v a l u e b e t w c e n l 4
and 17.
246
npsEARcH tN pHysropATHoLocy
oigl&
I ndex
7 days 12,2
!o
I
1.4
4.L
3.t
c
o { o
6
o
c
6
Eao
la
Ncrnal
3 hrs.
3,2
7o
Eo
Joo
22o
F t c . E 6 . S p e c t r a l a n a l y s i s ( . 0 1 i n e t h y l a l c o h o )l o f t h e t o t a l f a t t y a c i d s o b t a i n e d
from tbe body of rats irradiated with 1500r shows changes more manifcst around
270 mp. The oxalic index of the preparation is indicated and shows a parallel increase
with the changesin the curve.
RADtATTON
247
sacrificedafter the l3th day or which had died at any time, the index
showedvaluesbetweenl4 and 17. (Note 2/ Figure 85 showsthescresults
in the group of rats describedin this experiment.These changeswere observedwhen the same procedurewas repeatedin other goups of animals.
These experimentsclearly indicated that the quantity of conjugatedfatty
acids progressivelyincreasesin the days following the exposurein animals
lo
[\.*'
,"-
'iiradtated
o
an
o
e
-q
,O
E
(!
L
'a
Eo
Jto
tto
.t?o
frl"ue
tenqffir1l
,r<
Frc. 87. Spectral analysis (.01 in ethyl alcohol) of the fatty acids of the lotal body
of a mouse irradiated with l50Or, shows an increasein fatty acids with the absorption
correspondingto 270 mp., as compared with the control.
treated with one lethal dose of X-ray. Death occurred when the amount of
conjugatedfatty acid reacheda critical level equivalentto an oxalic acid
index value betweenl4 and 17. The spectralanalysisof fatty acidsof animals treatedwith radiation showedchangescorrespondingto the presence
of conjugatedisomers.These appearin the samplesof fatty acids obtained
from the entire body of theseanimals.(Figs. 86 and 87l Still more evident
werethe conjugatedtrienesin the fatty acidsof organs.Figures 88, 89 and
90 show the differencein such analysesas comparedto correspondingun.
248
xESEARcH rN pHysropATHoLocy
treatedcontrols.The presence
of conjugated
trienesappearsclearlyin the
characteristic
peaks.
The concept of a critical value for the oxalic index is supportedby
other studiesin which the same value is found in animalsdying after
adrenalectomyor after thermal, chemicalor traumatic statesof shock.
Even in animalsdying in superacuteshock,within 3 to 5 minutesafter
beingseverelyscaldedin hot water,the level of conjugatedfatty acidsis
higherthan in controls.
2c)-+-
0 . 0 1 2t n e t h y l a l c o h o l / e i h yal l c o h o l
Imariiated
,o
(o
c
o
q
vl
c
E6o
|9
t
Da
70
9o
,.o
3@
2to
waritnothr#Ft
22o
F r c . 8 8 . S p e c t r a la n a l y s e s( . 0 1 i n e t h y l a l c o h o l ) o f t h e f a t t y a c i d s o f t h e k i d n e y o f a
normal rat and of a rat irradiated 6 days previously with 1500r. The peaks characteristic for conjugated trienes are seen.
R^DrArtoN
249
0 . 0 1 2i n e t h y l a l c o h o l / e t h yall c o h o l
- lmadiated
Jo
t
a
I
t
to
c
o
t^
vt
E
tn
c
t>e
76
8o
9o
5oo
"?.r. 1.61fr
@r)t""
220
Frc. 89. The spcctral analyses (.01 in ethyl alcohol) of the fatty acids of the liver
of a normal rat and of a rat irradiated 6 days previously with 1500r shows the appearaoceof the cbaracteristicpeaks of conjugated trienes,
xEsEARcH rN pHysropATHoLocy
250 /
was to establisha radiation procedurewhich would producea standardized lesion.When radiationswereapplieddirectlyto the skin of animals,
the individualdifferencesin responsewere quite marked.Thesecould be
explainedin part on the basisof ageand particularlyof sex,the difference
,o
0.01%
i n e i I alcohol/ethyl
alcohol
,l r r adiated
(o
,
,
I
I
,
,
I
,
t
o 6o
an
v,
E
U'
c
(9
L 7O
i'i.,n, ,/'
r
\'
/""
ba
9o
ve Lenoth
(np;
F t c . 9 0 . S h o w s t h e s p e c t r a la n a l y s e s( 0 . 1 i n e t h y l a l c o h o l ) o f t h e f a t t y a c i d s o f t h e
lung of a normal rat and of a rat irradiated with 150016 days previously. Pcaks correspondingto coDjugaledtrienes are present.
RADIATION
,/
251
The problem of variability was satisfactorilyresolvedby radiating abnormal tissues,such as thoseof a wound, insteadof normal tissues.Standardizedlesionswere first producedand then irradiated.we usedthe following technique:an area of the skin on the back of male ratsweighingaround
200 grams was epilatedand, under ether anaesthesia,
a 2 cm.long incision
a
c)
E
q,
(o
P
L
r
'
x
T
Ccntrol
'H.r.
,
2
Oays
Ftc. 91. Fatry acid coniugation and irradiotion in yiv'o. The changes in the oxalic
i n d e x o f t o t a l f a t t y a c i d s o f r a t s s u b m i t t e dt o s u b l e t h a li r r a d i a t i o n ( 6 0 0 r . ) . O n l y a
temporary increasein the oxalic index of the fatty acids of the animats is seen, the
a m o u n l s n o t r e a c h i n gt h e c r i t i c a l v a l u e s .
252
R E s E A R C Hr N p H y s r o p A T H o L o c y
removedwith the help of the thread passedthrough the hole of the needle.
The retaining sutureswere also removed and the wound left open and undressed.
The length of time that the needle was left in place varied with the
amount of radium, the nature of the filtering metal, and the radiation burn
desired.we found that l0 mg. of platinum filteredradium had to be left
in place for about 90 houn in order to produce a standardizedulceration
that would last about four weeksbefore healing.The same effect was obtained when 25 mg. of monel metal filtered radium was kept betweenthe
lips of the wound for only two hours. when monel metal needleswere
used for only one hour, too great difterencesappearedbetween the ulcerations obtainedand the time necessaryfor their healing.A two-hour exposure causedan ulceration which usually required 4 to 5 weeks to heal in
control animals.lf the needlewas left in place for 3 hours, the ulcerations
were quite uniform but they required over two months to heal and more
than half of the wounds never healed.Failure to heal and more extensive
necrosisresultedfor periodsof exposurebeyond 3 hours.
Therefore,we utilized l0 mg. of radium in platinum for 90 hours in
one group of experiments,and 25 mg. of radium in monel metal for 2
hours in another group, in order to produce standardizcdulcerationsthat
would generally heal spontaneouslyafter 4 to 5 weeks.This technique has
been used in several hundred animals for various experiments. The fatty
acids of thesestandardizedradiation lesionswere studied.
Days after irradiation, the ulceratedlesionswere removed along with
about one cm. of surroundingtissueand submittedto analyses.It was always necessaryto use as many as 5 or 6 lesionsto obtain the quantity of
fatty acids needed for an oxalic index determination.The lesions werc
found abnormally rich in conjugatedfatty acids. Commonly, indices as
high as 4o-and in exceptionalcasesas high as 65-were found (Trnle
XVI) in comparisonwith 0 or 0.3 for normal skin with its subcutaneous
tissues.
Lipids and Radiation Burns
S) T'1t" apparanceof conjugatedfatty acids as an effect of radiation
has posed the problem of the role of theseabnormal fatty acids as intermediary agentsin the biological changesinduced by radiation. In trying
to solve it, we compared the eftectsobtained by administrationof conjugated fatty acids with those of radiation at different levelsof organization.
This study was facilitatedby consideringthe changeswhich take place in
R A D I A T I ( ) N
2s3
Trnle XVI
Ox,rlrc lNoex on Frrry Actos or Reorrrrox Bunxs
ElapsedTime
NormalSkin
Non-treated
wound
24 hours
48 hours
7 2 hours
6 days
2 hours
24 hours
4 8 hours
4 days
I week
')
weeks
3 weeks
Average
0.1
2.2
3.9
2.3
1.8
t.7
6.1
l3.9
l9.1
31.0
46.0
49.4
254
REsEARcH rN pHyslopATHoLocy
BADIATION
255
,,.,
,;,]]:,,i.
u
C
Ftc. 92. Lipids anct radiatian *'ounds. Radiation rvounds 5 weeks aftcr exposure to
l0 rngr. radium in platinum-for 96 hours. (a) Untrcated conrols: (b) treatcd tlnily
with I cc of a cod liver oil fatty acids l0{?. solution; {c) treated daily with 0.5 cc of
a 109* rolution of unsaponifiable lipid fraction cxtracted from human placcnta. The
lrcatmenl with fatty acids results in larger lesionsthan in controls, with no tendency
to hesl. Thc treatmcnt with the unsaponifiablc fraction lcads to a hcaling of the lesion
in rround threc wocks.
256
*ESEARCHrN pHysropATHoLocy
R^DrArroN /
257
258
RBsEARcH rN pHysropATHoLocy
monal secretion. The difterences between the effects seen in adrenalectomized and nonadrenalectomizedinadiated animals correspondsthus to
the adrenal responseto the systemicchanges.
In experimentson rats, we have shown that polyunsaturated,and especially conjugated,fatty acids inducechangesin the number of lymphocytes
a short time after their administration,and that this is followed by involution of the thymus, spleenand lymph nodes.This seemsto occur through
the intervention of the adrenals since it takes place to a greatly reduced
degreewhen the sameamountsof fatty acidsare administeredto adrenalectomized animals.The abnormal fatty acids seem to influencethe adrenals
and their responseelicits lymphopeniaand involution of lymphatic organs.
However, this indirect action through the adrenal glands is only part of
the story. Large dosesof the samefatty acidswill directly induce a certain
amount of lymphopenia and involution of the lymphopoeticorgans since
these changesalso occur when these fatty acids are administered in large
dosesto adrenalectomized
animals.
n) All of this researchindicatesthat two of the mechanismsthrough
which radiation acts upon the organisminvolve changesin lipids. In one,
the action is directly through fatty acids; in the other, as a responseto
thesefatty acids, anti-fatty-acidagentsintervene.The role of the adrenals
appearsto be still more interestingconsideringthe nature of the fatty acids
produced by radiation. As seen above, the conjugatedtrienes appear almost specificallyas a result of irradiation of mixturesof fatty acids.It was
also seenthat the corticoidsintervenespecificallyagainsttheseconjugated
fatty acids.This correlationseemsto representthe link betweenradiation,
conjugatedfatty acids and the adrenalresponse.(Ch. 6, Note l7)
Direct Action ol Radiation
o) In spite of the importanceof fatty acids and anti-fatty acids, they
representonly one part of the mechanismthrough which radiation acts.
The direct and indirect action of radiationon other constituentsalso must
be considered.Thc influence exercisedupon these constituentscan be
largely relatedto variouschanges.There is a quantitativerelationship,for
instance,betweeninductionof mutationsand the direct impactof radiation
on proteins.Changesin fatty acids also are the result of such a direct impact. It appearedinterestingto ascertainhow much and which of the pathological changesthat follow irradiation are due to the direct impact upon
lipids and how much to the impacton the other constituents.
The three kinds of biologicalactivity of radiation-through other constituents,through changed fatty acids. and reactional through anti-fatt)'
R^DrArroN /
259
260
xESEARcH rN pHysropATHoLoGy
lipidic effect by changing the nature of the radiation. The use of more
penetrating rays or of different corpuscleshas to be investigatedin terms
of the relationship betweeninfluenceupon fatty acids and the effect on
other constituents.It could be seenthat, in correspclnding
dosages,the less
penetratingradiations had a greater influence upon fatty acids than the
more penetrating.The fact explainsthe reductionof radiationburns directly
relatedto the interventionof fatty acids.Similarly,in a systemicprocedure,
such as teleradiotherapy,
the effecton other constituentsis reduc'edas compared with the direct influenceexertedupon the fatty acids.
It is possiblethat radiationsusing neutronswould induce an increascd
direct impact on other constituentswithout a correspondinglyincreased
effectupon the systemicfatty acids,The skin effect,which is minimal with
theseradiations,would indicatelittle interventionof fatty acids.
The unequalpart playedby lipids at the different levelscan be utilized
to obtain variationsin radiation eftects.If effectsupon the lowest levels of
the hierarchicorganization,such as upon histonesand basic amino acids,
are desired,radiation could very well be the tool to be chosen,becauseof
the small amountsof lipids prcsent at these levels. If the influencecould
be limited to such action, radiation could be consideredideal for such
therapeuticeftects. Unfortunately, this is not possible even when very
penetratingradiation is used, and the eftect of radiation upon lipids still
constitutesone of the principal factors which must be consideredwhen
radiationis usedas a therapeuticweapon.
Thus radiation is not the ideal meansfor affectingthe subchromosomal
level, in spite of the fact that it may, througlr its effect upon proteins,have
a profound influencebelow this level. Its ability to causea conjugationof
fatty acidsrepresentsthe seriousobstaclcto its use. In view of this. the
effectof radiation upon lipids actuallycan be consideredas an undesirable
epiphenomenonwheneverthe purposeof the therapy is to achievea local
effect at the lowcr levels. Frequently, the changeswhich require discontinuationof radiationtherapycan be recognizedto correspondto abnormal
local or systemicmetabolismproducedby the abnormalfatty acids.
It must, however,be recognizedthat the appearanceof abnormal fatty
acids has some advantageseven upon protein effects,since indirectly they
can make local tissuesmore sensitiveto radiation.We have previously
noted that abnormalfatty acids causcchangcsin the tissueand cellular
metabolismwhich lead to local alkalosis.This local pH changemay have
favorableresultsby acting upon the amphotericproteinsand by increasing
the positivelychargedmemberswhich apparentlyare the only ones sensi-
R^Dt^rtoN /
261
262
xESEARcH rN pHysropATHoLocy
R^DIATION
263
C H A P T EI RI
P R O B L E M SI N C A N C E R
NT
PROBLEMS tN
CANCER
265
266
xesEARcH rN PHYsToPATHoLocY
nant cancer in the clinically frightening sense.On the other hand, the
processeswhich are addedto a noninvasivephaseform of cancr and turn
it into the invasive,tissue,organicor systemicphase,have no characterof
specificity. Similar growth changes,or the appearanceof lipidic predominance, which representadded factors are seen in many other conditions.
By using them for diagnostic purposeswe will not recognizn,the cancerous
condition but only nonspecificinterveningfactors. While these factors are
responsiblefor the changes,malignancydevelopsonly when, and because,
thesefactors operateon already abnormalentities,i.e., cancerousentities.
This explainsthe nonspecificityof many proposedtestsand the misleading
positive results obtained in conditions such as pregnancy where one of
theseadded factors, (active growth processes)is always present.
A test for cancer, to have clinical value, would have to indicate two
things: one, the specificearly changewhich is widely distributedbut representsthe cssentialcondition for the potential developmentof malignancy;
and, two, the concomitantprcsenceand concomitantoperationof the nonspecific factors which can cause the actual developmentof malignancy.
This kind of diagnostictest undoubtedlywill come from further systematized study of biochemicalchangesinducedby the simultaneousaction of
the two groups of factors.
Immunological studies reprcsentan approach of value for diagnosis.
The different phasesof cancer can be interpreted,in the final analysis,to
correspondlargely to the interventionof the defensemechanismat different stagesat the different levels.As mentionedabove, a changein a phase
resultsalso from a changein the defensestageat the respectivelevel. We
have seenthat the immunologicalaspectof cancer cannot be understood
without acceptinga relative independenceof the levels in their different
stagesof defense.This view explains somc seeminglyparadoxicaloccurrences.
Cancerouscells are frequently found circulating in the blood yet this
does not indicate generalizedcancer. While the organism defends itself
successfullyat thc systemiclevel againstcancerouscells, the cancer can
still progressat the lower level of the tissues.The loss at this Iow level of
an effectivedefense,principally primary or allergic,which is still persistent
at the systemiclevel, explainswhy the cancerouscells invade the tissues.
A test indicating the presenceor absenceof any immunologicalreaction
would consequentlyhave value only when related to hierarchic levels. It
must furnish indicationsonly of what is happeningat a specificlevel. The
nature of the immunologicalreaction in cancer is also different from the
reactionin other conditions.Defensecapacity-natural defensecapacity-
PROBLEMstN cANCER
267
268
RESE^RcH tN pHystop^THoLocy
ous cells which get into the blood. This is not true at the level of the
interstitial formations, that is, at the tissular level, where such defense
means are failing. The real danger during surgeryconsequentlyis not so
much the presenceof cancer cells in the blood, since the blood can still
take care of them, but the spreadof thesecells at the tissular level where
the defensecapacityhas beenlost. and where a cancerouscell consequently
has every chancenot only to remain alive but also to grow.
The independenceof the defensemechanismat different levels also
must be takeninto accountin explainingthe differences
in the eventswhich
follow the appearanceof a spontaneous
tumor and those which occur after
cxperimentaltumor transplantation.
EXPERIMENTAL
CARCINOGENESIS
i,ffi#l:*
PRoBLEMS rN c^NcER
269
aspect,carcinogenesis
no longer can be acccptedas a simple processoccurring in the cells, but must be regardedas a successionof organizcd
processes.
This becomesstill more interestingwhen it is realizedthat changesin
the constituentsat the lowest levels of the organizationcan occur on a
statisticalbasis,that is, independentlyof the direct interventionof external
agents.As thesechangeshave to be developedfor many successivehierarchic entities, it takes a certain time for them to be realized. This would
explain why most cancers appear after a certain age. Cells with cancerous
nuclei, i.e., ln the noninvasivephase,frequentlyare present,in older people, in many organs without producingclinical manifestations.Conceptually, in order for an agent to be considered a successfulcarcinogen, it
must act upon these noninvasive entities to such an extent as to change
them into invasiveones. It can thus act upon entities which have already
progressed,by themselves,far enough in the hierarchicdevelopmentof a
cancerous processand have arrived at the noninvasive phase without any
manifestation.The excessivelength of time necessary,even for the most
active agents,to induce invasive cancer would suggest,however, that more
than a simple passagefrom an already existing noninvasive cancer into
an invasiveone is involved. A plurality of changesmust be induced,some
or all at levels below the cell.
We are inclined to favor this last hypothesiswhich obliges us to consider that a carcinogeninduceschangesat differentlevelsof the organization. It is supportedby a seriesof facts. In addition to having the capacity
to induce invasive tumors, carcinogenic agents also induce precancerous
lesions which correspondto cancerousentities below the invasive phase.
Cells with abnormal nuclei or with only abnormal chromosomesare almost
constantly seen in induced carcinogenesis.
Even agents which produce a
high proportion of invasive cancer consistentlyinduce such changes at
Iower levels as well. For carcinogenswhich induce a low proportion of
invasive cancers,the effects often appear to stop at Iower levels. Such
activity at subnuclearlevels of the organizationis seen in the capacity of
most of the carcinogensto induce mutationsand monstrosities.
In the conceptof hierarchicorganization,mutations are consideredto
result from changestaking place at the gene level, with lower levels left
unaffected.Monstrositiesresult from changesat the chromosomelevel.
with mutationsand monstrositieshas led us
Comparisonof carcinogenesis
to consider that cancerouschangesbegin at levels much below those involved in mutations and monstrosities,possiblyat the nucleo-proteinlevel
or. even below. The complex cancerouscondition to which the invasive
270 /
x z s E A R c Hr N p H y s r o p A T H o L o c y
P R O B L E M Sr N C A N C E R
271
carcinogenicproperties.This conceptwas partially modified and amplified by Pulman,Dawdeland their co-workers(44) who have shown,by
quantumanalysisof variouscarcinogens,
that the densityof the a electrons
is increasedin certain preferredregionsof the molecules,the K regions.
They showedthat, whenelectrondensities
exceed1.292eat theseregions,
the substances
have carcinogenic
properties.Figure 95 showssuch a K
region.
o)
i-=
\z
\,2
CHs
9:l0 Dimethyl
| :2:7:8 Dibenzanthracene
Flc. 95. The regions K in carcinogenicmolecules.
From our point of view, a tentativelyinterestingaspctof this condensation of rr electronslies in two facts: the presencein some carcinogen
moleculesof more than one such K region, and the presenceof different
valuesfor theseK regionsin differentmolecules.It would be the presence
of more than one K region in the same molecule which would result in
interventionin more than one processand thus contributeto plural activity.
Further analyses,however,suggestedthat the condensationof z electrons in K regions would represent only one of the factors that would
induce activity in theseagents.We have identifiedanother energeticfactor
in the presenceof two atoms having the same electrical sign and being
bound togetherwithin the molecule.
Twin Formation
We have considered the existenceand importance of these "twin formations" as indicationsof energeticactivity in the course of studies on
electronicmoleculararrangements.In a molecule,an alternationof successiveatomsresultsin part from the alternatingpolarity of theseatomswithin
a molecule and in part from the opposite charactersconferred upon the
272
R E S E A R c Hr N p H y s r o p A T H o L o c y
two carbon atoms when they form acetic acid, an important precursor in
biologicalsyntheses.It is through alternatepolarity that an induction effect
of an energeticcenter in thc molecule propagatesitself along the chain.
The presenceof any energeticcenter in the moleculerepresentedby polar
groupsor a lateral chain, for instance,will enhancethis alternalepolarity.
When one or more such inductiveetTectsare propagatedthrough the chain,
two adjacentatoms may be found to possessthe same electrical sign for
their chargeor ionoid character.The twin formation which results representsa center of increasedmolccular reactivity. This reactivity can be so
intense as to lead to breaking down of the molecule, something which
occurs often in inorganic substances.
This has led Pauling to believe that
this condition,called "adjacentchargeru|e," cannot exist.
"Pauling has pointed out that the murual potential enerry of two electrical chargesof the samesign is so high that a canonical structurehaving
net residualchargesof the same sign on any adjacent atoms would have
too high an energy level to contribute appreciably to the real molecular
structure." So notes William A. Waters in "Physical Aspects of Organic
C h e m i s t r y . "( 4 5 )
The form suggestedfor nitrogenperoxide (N.On), (Fig. 96l would
appear to be impossiblebecauseof the high energy developedat the two
positive nitrogens.
NitrogenPeroxide
Frc. 96. The existencc of nitrogen peroxide molecule is prevented by the high energy developed at the two adjacent positive nitrogens.
However, the forces that exist in most of the organic molecules are
much weaker, so that the resulting"twin formations," although energetically potent. arc not strong enough to induce the breaking down of the
molecule.Consequently,they would exist and rePresentimportant energetic
centers.
P R O B L E M Sr N C A N C E R
273
we have studied a number of carcinogenicagents,seekingtwin formations. Analysisof the ionoid characterof the carbonsof the methylcholanthrene molecule revealsthe presenceof twin formations which could be
localizedat various points of the molecule.Figure 97 shows the energetic
aspectof methylcholanthrene
and the ionoid characterof its carbons.It is
the presenceof the cyclopentanegroup in the molecule that induces the
samesign in two adjacentcarbons.The presenceof the methyl group would
determine the electrical character of C2e and consequentlythe srccession
of alternate signs. On the other hand, the double bonds will determine the
probable localization of these twin formations in the molecule at the K
formation itself, that is, at C5 and C6.
Hsc
1)
20-Me
t hyI choI anthrene
Ftc. 97. The energeticaspectof methylcholanthrene.with twin formations.
Twin formations can be found in many carcinogens.It must be emphasized,however,that unequal energeticvaluescan be recognizedeasily
for difterent twin formations and would explain differencesin their activity, a fact which would confer possible plural properties upon this group
of qualitativelysimilar energeticformations.
Another aspect of the relationship between these formations and carcinogenesisappearsto be even more interesting.While no twin formations
can be found in severalagents,the formations are presentin the substances
resultingfrom metabolism of these agentsin the body. The relationship of
twin formation to carcinogenicactivity can be suspectedwhen such changes
appear simultaneouslywith carcinogenicity.
For example,no twin formation occurs in 2-naphthylamine,(Fis. 98)
whosedirect carcinogenicityis questioned.but such a formation appearsin
heterocyclic3:4:5:6 dibenzcarbazole,
one of its intermediates(46), which
is known for its carcinogenicproperties.(Fig. 98bis) This is also true for
274
nESEARcH rN pHysropATHoLooy
NHz
(a)
2-Napht
ylamine
Ftc. 98. No twin formations exist in 2-naphthylamine.
(b)
3:4:5:6 Dibenzcarbazole
F r c . g E b i s .A t w i n f o r m a t i o n a p p e a r si n t h e i n t e r m e d i a t e3 : 4 : 5 : 6 d i b e n z c a r b a z o l e .
PROBLEMS IN
CANCER
275
NHz
Z-Aninofluorene
F l c . 9 9 . A t w i n c a r b o n g r o u p i s p r e s e n ti n a m i n o f l u o r e n e .
(-)
cHg
N: l'l
cHg
4
Dimethylamino
azobenzene
F l c . 1 0 0 . A t w i n f o r m a t i o n i s p r e s e n ti n 4 - d i m e t h y l a m i n o - a z o b e n z eantet h e l e v e l o f
the azo bond.
ti.,v:.Si:"Sffi&Sl
276
n E S E A R c Ht N p H y s r o p A T H o L o c y
<\
N
(+)
(+)
2:2' Azonaphthalene
Frc. l0l . 2:2' Azotaphtbalenehas only a slight activity.
2:2'
l: I'
Di a mni o
dinaohthyl
An interestingaspectis furnishedby urethaneand other estersof carbamic acid.Figure 106 showsthat no twin formationscan be seendirectly
or through a changein the molecule.This accordswith these substances'
lack of capacity,noted by many authors,to induce cancerouslesionsor
e v e nt u m o r s .( 5 8 , 5 9 , 6 0 ) O n ( 6 1 ) r e l a t e sl e s i o n sp r o d u c e db y c a r b a m i c
PROBLEMS tN
CANCER
277
/C",,,
\.r. ,*,
CH: CH
lott
D i e t h y l si ltb e s t r o l
F r c . 1 0 4 . A t w i n f o r m a t i o n e x i s l si n d i e t h v l s t i l b e s t r o l .
278 /
x E S E A R c Hr N p H y s r o p A T H o L o c y
Triphenyl-ethylene
F r c . 1 0 5 . T h e p o s i t i o no f t h e t w i n f o r m a t i o n i n t r i p h e n y l - e t h y l e n e .
NHe-C0-OCzHs
Urethan
F r c . 1 0 6 . U r e t h a n h a s n o t w i n f o r m a t i o n a n d a p p a r e n t l y - a c c o r d i n gt o m a n y a u t h o r s
- n o d i r e c t c a r c i n o g e n i ca c t i v i t y .
PRoBLEMSrN CANCER
279
ically active,and Haddow has shown that activity is presentonly if hydrolysis is sufficientlyhigh. (62) The inequalityof hydrolysisin different
memberscan be related to the influenccexertedby the radical bound to
the nitrogen.It seemsthat the presenceof a strongercnergcticcenter,as
it appears in positively or negativelycharged atoms bound to the cyclic
radical, reduces the dissociationof the chloroethyl group. Generally.
nucleophilicgloups would retard the dissociation.
Sufficientevidenceexists
to show that biologicalactivity follows the elimination of the chloride ion
and the appearanceof a carbonium ion as a reactive intermediate.A
further passageinto the ethyleneimonium
ion, consideredmore stableand
R *a"'
'cH2
cHzcl
c+z ct
NitrogenMustard
F l c . 1 0 7 . T h e n i t r o g e nm u s t a r dd e r i v a t i v e s .
280
xEsEARcH rN pHysropATHoLocy
zCHz-CHz-CL
C H-sN <\ - C H z - C H z - C L
I
nCHz
*ct-
CHr-r'r.(CH.
\r*,
- c H z- c t
+Hz0
J
-.CHz-CHr-oH
cHs- N< - C H . - C H r - c L
, L H r - C H ro H
/
CHs- u "(-Cn.
\l ' C H r
+ Ct-
+ HrO
/.'CHz
C H s- N . _ _^ . .
- CHu - OH r+l
-CHr -CHr - 0H F)
Dielectrophilic
'Ihc
Fto. 108.
c h a n g e sc l c c u r r i n gi n t h e n i t r o g e nm u s t a r d l e a d s t o e t h y l e n e i m m o n i u m
i n w h i c h a n e n e r g e t i ca s p e c ts i m i l a r t o r h a t o f a t w i n f o r m a t i o n i s p r e s e n t .
P R O B L E M Sr N C A N C E R
2gl
in methyleneimine
l:3:5 triazine,(Fig. l,r0l Through the influenceexerted
by these centers,the ethyleneiminegroup can have its carbons charged
sufficientlyto bccome a dielectrophilicformation. The possibilityof a reactive intermediate and a more stable electrophilic form thus appears
common to the two groups,mustardsand ethyleneimines.
;>. a=-<-;
\z
N
H,cArr,
Triethylenimine
2-4-6-Triazine
Flo. 109. Ethyleneimines are active carcinogens,probably related to their energctic
aspectwith a formation energeticallysimilar to the twin formation.
Eporide Cucinogens
A similar condition is also found in the epoxide carcinogens.carcinogenic activity has been recognizedfor substanceshaving two epoxide centers in close proximity in the molecule. The epoxide center by itself can
leadto a formationsimilar to that of carboniumion, as seenin Figure 108
and thus to the same formation found in mustards and ethyleneimines.
The analogy goes still further. The energeticcenter appears insrrfficientto
accomplishbiologicalchangeswithout an inductiveactivation.In the caseof
epoxides, this is usually brought about by another similar epoxide group
in the samemolecule.
As no carcinogenicactivity has been found in substanceswith only one
epoxidecenter or with two epoxidecentersfar apart, the inductive centers
seemto be of primary importance.The two energeticcentersforming the
epoxide g'oup, similar to those of the ethyleneimines,do not alone appear
282
,/
xEsEARcH rN pHysropATHoLocy
NOz
i)
N
H,cAcH,
2 : 4 D i n i tr o p h e n y l
Ethylenimine
F I c . l l 0 . T h r o u g h t h e i n f l u e n c ee x e r t e db y t h e n i t r o g r o u p u p o n t h e e t h y l e n e i m i n e ,
t h e i m i n e g r o u p o f 2 : 4 d i n i t r o - p h e n y l - e t h y l e n e i m i nbee, c o m e sd i a l e c r r o p h y l i c .
moleculeis metabolized,with the differencethat the two CHr in this instance seem to come originally from other chains. (Fig. I I I ) For the
it is possiblethat a similar processoccurs during the
methylolamides,
changesthat take placein the organism.
Some corroboration can be found in the fact that two forms can be
observedin theselast groupsof carcinogens.One is electrostaticallyactive;
that is, it has a certain ionic character.The secondhas a dual electrophilic
activity which can be relatedto a twin formation with nrolecularreactivity.
Thus, twin formation,with its specialreactivity,appearscommon in
many carcinogenicagents.To be biologicallyactive,the twin formation has
to be sufficientlystrong and this is insured by an induction effect exerted
PRoBLEMS
rN
CANCER
283
by other formationsin the molecule,such as double bonds in parallel position or polar groups.A twin formation as energeticcenter in the molecule
would exert a molecular field effcct. It would thus representa center of
molecularreactivity which has to be consideredas such in the analysisof
plural activity.
Ar.N{
- . C H z - C H z 0 S0 2 ' A r
RCH2+
t
\cHr c H 2- o s o 2 .A r
-0502' Ar
R C H 2 ++
Sulfonoxyalkane
-0302'
Ar
ReactiveIntermediate
F I c . t l l . C h a n g e so c c u r r i n g i n s u l f o n o x y a l k a n c sl e a d i n g t o t w o a c t i v e C H r c e n t e r s .
SynjugatedFormations
The study of variouscarcinogenshas permittedus to recognizeand
relate to complexcarcinogenicactivityanothercnergeticinfluenceexerted
by two or more doublebondswhen prcsentin a parallelreciprocalposition
in cyclic molecules.This led us to the conceptof "synjugatedformations"
with 2, 3, 4 or more suchparalleldoublebonds.
In studyingmethylcholanthrene,
one o[ the most potent of the known
carcinogenicagenLs,the curve of its absorptionin ultraviolet light was
considered.This curve is shown in Figure | 12. The place and form of
the peakscould be interpretedin a peculiarway when conjugateddouble
bond formationswere considered.In thc curve of methylcholanthrene,
we
portions
that corrcspondto an inverseof the curvesobcould recognizr
tained from variousconjugatedpolyenes.Furthermore,the curve obtained
through the spectralanalysisof methylcholanthrene
can be consideredto
have high similaritiesto the inverscof the curveof a mixtureof conjugated
polyenes.Figure I l3 showsthe spectralanalysisof conjugatedcod liver oil
f a t t y a c i d s ,w h i l eF i g . l l 4 s h o w st h e i n v e r s ec u r v eo f m i x t u r eo f c o n j u g a t e d
fatty acidsof cod liver oil in which conjugateddi-, tri-, tetra-,penta-and
hexaenesare identified.Figure I l5 shows the comparisonbetweenthe
and the inverseof the peaksof the mixture.
curve of methylcholanthrene
We were thus led to consider the conceptualinterpretationof these
curvesin termsof the specialrelationshipthat existsbetweendoublebonds
in the same molecule.In the classicalconcept,two double bonds are consideredconjugatedif two of their carbonsare joined by a singlebond. ln
the zig-zag representationof aliphatic molecules,the conjugated double
r,iti{
284
RESEARcH rN PHYSToPATHoLocY
c
o
I
c
L
Y a v eL e n g t h( n 1 )
cule. Two or more double bonds in a cyclic molecule would thus realize a
similar kind of energeticformation when parallel, and would do so independentlyof the number of the singlebondspresentin-between.(FiS. 1I6c)
For didactic purposes,we have applied the term "synjugated" to energetic
formations resulting from parallel double bonds separatedby more than
one single bond.
- , , r - : r . $ : : , :':+--"1ar.i:i:
{1$.
:1 .i"r: i..;itri. ,
I riilL
P R O B L f : M S I N
C A N C E R
285
c
cl
; $
tn
E
o
C
.E
L
*60
br
mpp))
f i a v e L eennqqt thh ( m
F t c . I 1 3 . T h e c u r v e s h o w st h e s p e c t r a a
l n a l y s i so f r h e m i x r u r e o f c o n j u g a t e df a t t y
a c i d sw i t h m e m b e r s h a v i n g f r o m 2 t o 6 d o u b l e b o n d s . a s o b t a i n e d b y t r e a t i n g c o d
liver oil fatty acids with KOH.
286
R E S E A R c Hr N p H y s r o p A T H o L o c y
c
ct
o
tt
E
tt
c
(!
L
a.a
PRoBLEMS rN CANCER
287
evenpenta-synjugated
formations.According to the concpt of plural activity in carcinogenesis,
synjugation,while not indispensablefor carcinogeneticactivity,would representone of the factorsthat can make it possible.
Togetherwith the condensationof the electronsin the K regionsand
".
the presenceof polar groups, the twin and synjugatedformations would
?to
o
(-
-c
c
,'
,
.I'
l\
, l
Eta
|
I
-c
269
I
L -
60
l ' .
J \
c
o
'
it
234
, ' I
'izo
c
(g
L
}Q
Joo
280
250
288
REsEARcH tN pHysropATHoLocy
a
Frc. l16. Conjugation and synjugation. ln the aliphatic chain (a) the presence of
single bonds between the double bonds induce the parallel position of double bonds.
It is this parallelism, which through thc reciprocal influence cxertcd, induccs thc
energctic characteristicsof the conjugated formations. Io the benzene molecule (b)
whcre thc double bonds, although separated by single bonds, arc not paraltel, thc
lack of this parallelism explains the lack of the properties charactcristic to thc conjugated formation. Tbe parallelism when present in cyclic molecules (c) realizcs thc
"synjugated" formations.
sult of difterences in their energetic spectra. The di.fterencesare consequently qualitative as well as quantitative. From this viewpoint, it is
possiblethat the great carcinogenicactivity recognizedfor some substances
would correspond to the presencein them at once of a great number of
energeticfactors.
The study of the correlationbetweenthe presenceof various energetic
centers and carcinogenesishas been facilitated by relating carcinogenic
changesto levels of organization.Taking place at different levels, the induced processes
can be seento correspondto an entire seriesof manifestations which, while present also in invasive cancer, often can be recognized
PRoBLEMSrN cANcER
289
Tetrasynjugat
ed
p;
HS
/
.r'
Trisynjugat
ed/----/-'- J
/'
Eisynjugat
ed
ion in 20-Methylcholanthrene
Synjugat
F I c . l l 7 . T h e p a r a l l e l p o s i t i o n o f t h e e x i s t i n gd o u b l e b o n d s i n m e t h y l c h o l a n t h r e n e
c o r r e s p o n d st o a b i - , t r i - , a n d t e t r a s y n j u g a t i o n .
Tl l n ai
.a--
\
-\
cHs
\ i
il-l
r ll
\ / l
{
I
f-\ I
,
t
.
/
t
l -
lt /
'l r l r
-'\
t
T w o K R e g i o n s+ .
T r oT r i n F o r m a t i o n s
[tneTrisynjugated
Bond
Tro Tetrasynjugated
Bonds
L'{r H :
/
Tetra
-V
Tetra
9 : 1 0D i m e t h y ll : 2 : 7 : 8 - D i b e n z a n t h r a c e n e
F r o . l l 8 . ' f h e e n c r g e t i cp i c t u r e o f 9 : 1 0 d i m e t h y l . l : 2 : 7 : 8 d i b e n z a n t h r a c e nseh, o w s
t h e p r e s e n c eo f t w o K r e g i o n s ,t w o t w i n n e g a t i v ef o r m a t i o n s ,o n e t r i s y n j u g a t e db o n d
and two tetrasynjugatcdbonds.
29O /
n E s E A R c Hr N p H y s r o p A T H o L o c y
PRoBLEMS
tN
CANCER
2gl
292
xEsEARcH rN pHysropATHoLocy
PRoBLEMS lN
cANcER
293
294 /
B , E s E A R c Hr N p H y s r o p A T H o L o c y
PROBLEMS lN
CANCER
295
(80), only partial inactivation can be consideredto occur since electrophoresis,repeatedpassage,or even dilution restorespathogenicity,The
restorativefactor can be of varied nature.Casesin which pathogenicityis
restoredby a nonviral agent-activation of the virus of swine influenzain
the presenceof Hemophilusinfluenza(81), for example-are most revealing.
Virus and the Host
This conceptof plural activity cxplainsthe rclation betweentumorgenesisand destructioninduced by viruses.Often "neoplastic" infection
and "destructive"infectionare inducedby the samevirus. (82)
The herpes virus thus induces necrotic lesions in the chick embryo
whenintroducedin early stages,but if the cmbryo is more developed,the
samevirus producesproliferativechanges.(83) The myxomavirus induces
more proliferative lesions if attentuatedthan does the unchangedvirus.
(84) Under specialcircumstances,
such as in oldcr animals,shecp pox
virus inducespapilloma insteadof pustular infection (85). It must be
remarkedthat thesedifferentresultsare not limited to viruses;they occur
with radiationor even with other infectiousagents.(86) Bartonellabacilliformis,which inducesoften-lcthaloroya fevcr, seemsto be the causeof
"verruga peruviana," a fibroangiomatoustumor often seen in subjects
recoveringfrom the acute disease.
The differencesin activity of thc same virus appear to be related to
the age of the host. Generally,youth of the host increasesthe virus' capacityfor acting at more levels.The virus can produce lethal destructive
disease
in young animalsbut only a neoplasticrcsponsein adults,as seen
for the fibroma virus in rabbits.Furthermorc,the neoplasticresponsealso
occursin young animalsbut only if a small amountof virus is inoculated,
or if an attenuatedvirus, such as a long-storedone, is used. (87) This is
clearin the caseof the Rous sarcomaand other chickcntumors.
When injected into very young animals. Rous sarcoma and other
chickentumor virusesproduce a hemorrhagiclesion (88) but they will
inducetumors in adult animals.The dcstructivccffect can bc repeated
with repeatedpassages
of the virus in very young animalsbut in adults
eachpassageproducesthe ncoplasticresponsc.This is also true for some
strainsof lymphomatosisvirus (89) which induce tumor formation in
adultsand necrotizingprocesses
in young animalsor embryos.It is also
true for the virus of neurolymphomatosis
(90), and of gliomas. (91)
Theseviruses,althoughselectivefor thc nervoussystem,induceinflammatory or neoplasticlesionsaccordingto the age of the infcctedanimal.
;1,,,,i1#ff$ii#ffiffi,($
296
REsEARcH rN pHystopATHoLocy
PRoBLEMS tN c^NcER
297
298
xESEARcH rN pHysropATHoLocy
PROBLEMS IN
CANCER
299
300
REsEARcH rN pHysropATHoLocy
PROBLEMS tN
cANcER
301
(2e4)
302
n EsEARcH rN pHysropATHoLocy
PRoBLEMS rN cANcER
lOl
304
x E S E A R C Hr N p H y s t o p A T H o L o c y
P R O B L E M S
I N
C ^ N C E R
305
306
xESEARcH rN pHysropATHoLocy
PRoBLEMS tN C^NcER
3O7
,,,giw'*ffi,
308
R E S F - A R c Hr N p H y s r o p A T H o L o o y
P R O B L E M Sr N C ^ N C E R
309
t22\
In summingup this researchin a recentpapr (123), the following
conclusions
couldbe reachedby LcShan:
l. There seernsto bc a correlationbctweenthe existenceof neoplasticdiseaseand the persistence
of certaintypesof psychological
situations.
2. The most consistentlyrepnrted.relevantpsychologicalstate has
beenthe lossof a major cmotionalrelationship.Often the psychic
state resulting from this loss could be traced to a period shortly
beforethe first notedsymptomsof cancer.
3. There appearsto be somerelationshipbetweenpersonalityorganization and the evolutionof the cancerouscondition.
4. There may be some relationshipbetweenpersonalityorganization
and the type or location of a cancer.
It would seemas if future researchin this area,to bc as usefulas possible,must focusupon the chemicophysiological
changeswhich resultfrom
variationsin psychic statesin general.It is highly probable that these
changesare mediatedthrough the endocrinesystem.Through the linking
of psychic stateswith hormonal changes,we may be able not only to
integratepsychologicalfactors with the many other factors influencingdevelopmentof cancer,but also to relatcthcm to certainlevelsof organization. It may be possibleto establishthe relationshipof psychologicalfactors
to other influenceswhich favor or even inducepassageof cancerfrom one
phase to another. This may prmit a more complete understandingof
cancer and help in finding new points at which some therapeuticvalue
might be expectedfrom psychological
intervenrion.
The relationshipbetweenthe adrenalsand psychicstateson the one
hand,and betweenthe adrenalsand the lymphaticsystemon the other hand.
couldexplainwhy the inlluenceof a psychologically
unresolvedproblem is
most evident,among all cancerousconditions.in lymphomas.LeShanhas
been able, by analyzinga sizablenumber of these lymphoma cases,to
310
./
nEsEARcH rN pHysropATHoLocy
C H A P T ELR2
PHARN,IACODYNAIUIC]
ACTIVITY
TT
we have investigated
l[* ou* REsEARCH,
the pharmacodynamicactivity
of a seriesof substances
in terms of thc physiopathological
conceptsdiscussedabove: the level of organizationat which thcy act, the dualistic
natureof their activity,their relationshipto body constituents,
especiallyto
lipids, and the changesthcy induce in the dcfensemechanism.We will
limit ourselvesin this presentation
only to thoscof the substances
investigated which are of therapeuticinterest.
The dualistic concept has pcrmitted didactic separationof agentsinto
two groupswith antagonisticproperties.In the last analysis,this separation
could be related to the two fundamentaltendenciesin nature, homotropy
or heterotropy.For inorganic agents,this criterion appearsto be directly
relatedto the elementspresent.However, for organic agents,especiallyfor
lipids and lipid-like substanccs,
this simplecritcrionis lessvalid. Another,
the positive and negativeelectricalcharacterof their active polar groups
could be used. It must be emphasized,however,that some of these polar
goups show either positivc or negative properties depcnding upon the
medium in which they work. An alcohol,for cxample,can act as an acid
under special circumstances,in which case it forms metal alcoholates;or
it can act as an alcoholforming esterswith acids.For this reason,in conwe havc limited the sphereof changesstudiedto
sideringthesesubstances,
thosecorrespondingto the conditionspresentin biologicalentities.
Since, as seen above, substanceswith negativc polar groups actively
while those with positivepolar groups conintervenein various processes,
trol tbe activity of the first, we will start this presentationwith the former.
3ll
312
xESE^RCHrN pHysropA't-HoLoGy
Acids
SaturatedFatty Acids
The lipoidic character in this homologousseries starts with the five
carbon valeric acid, although caproic acid is the first lipoidic member
found in animals.The role of the saturatedfatty acids in organizationis
largely related to the activity, as caloric metabolites,of the memberswith
even carbon numbers.These saturatedfatty acids are absorbed,circulated
and stored as triglycerides,and it is also this bond to glycerol which ap
parentlyfavorstheir biologicalrole as caloricmetabolites.
As previously noted, only the memberswith a relatively short chain,
lcss than 12 carbons, can be directly metabolizedthrough Knoop beta
oxidation.Longer chain saturatedfatty acids undergo a breakdown before
further metabolic degradation.Members with l6 and more carbons also
play a limited constitutional role, usually together with an unsaturated
member, when bound to glycerophosphoricacid to form phospholipids.
Even when frec-that is, whcn their polar group is not neutralized-they
do not have a manifestfunctional role and, consequently,show no important pharmacodynamicactivity.
The higher fatty acids exert no dcmonstrableinfluenceupon microbes
nor upon viruses such as bacteriophages.
No influencehas been observed
upon monocellularorganisms,cells or tissuesin vitro. No changeswere
seenin the respirationof liver slicesor of Sa 180 ascitescancerouscells in
the Warburgmicrorespironreter.
Similarly,no influencecan be seenon red
blood cells nor on leucocytestreated in vitro. No changesin the chloride
P H A R M A C o D Y N A M T CA C r r v t T Y
3 1 3
;,lr1[arr,
Fil..:'S
314
,/
xESEARcHlN
pHysropATHoLocy
dermotropeviruses.Injected subcutaneously
in rabbits, oleic acid induces
in the skin, at the site of injection a zone of low receptivity for smallpox
virus. Oleic acid has a reducedeffect upon microbes,causingthe appearance of some gram negativeindividual forms of Bac. subtilis, for instance.
Mixed directly with blood, oleic acid induceshemolysis.when plasmais
treatedwith this acid, and then mixed with red cells.the influenceexerted
upon red cells becauseof the small amount of acid fixed upon plasma, is
reduced. Although olcic acid may influence the pH of the second day
wound crust, causingan increasedlocal alkalosis(Fig. II9), its influence
0leic actd
F t c . l l 9 . T h e s e c o n dd a y w o u n d c r u s t p H f o r o l e i c a c i d
s h o w st h e c o n s t a n tp r e s e n c eo f a c h a n g et o w a r d m o r e a l k a l i n ev a l u e s .
upon pain is almostnil. Little or no changeis scen in standardizedradiation woundsin animalstreatedwith the acid. A limited influencecan be
observedon tumor grorvthby using thc techniqueof dipping transplants
gencrations.Usc of a 1094.solutionof oleic acid in tricap
for succcssive
gcncrations,
impairsgrowth of
rylin bcforegrafting,rcpeatcdfor successivc
after thc sixth or sevcnthgenerationin
Ehrlich mouse adenocarcinonra
some experiments,even later in othcrs.Negativepassagetakes place betweenthe cighth and tenth graft. Under the samecondition,very little or
no changesare noted in other tumors,and no changesare seenin tumors
in animalstreatedwith this acid even thoughchangeshave been reported
of cells of differentorgans,treated
by some authors, (124) Suspcnsions
of oleic acid and injectedin animalsof
in vitro with colloidalsuspcnsion
P H A R M A C O D Y N A M T CA C T T V T T Y
315
,-t.f
316
R E s E , ^ R c l rr N p H y s r o p A T u o L o c y
PHARMACoDYNAMTC ACrlvrTY
317
rfll:'-.
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.ji
$ffi
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318
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RESEARcH IN
PHysIopATHoLocy
:6
a
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aa
lave Lengthfnp.;
F l c . 1 2 0 . S p e c t r a la n a l y s i so f r a t l i v e r f a t t y a c i d sa f t e r c h e m i c a li s o m e r i z a t i o ns h o w s
t h e p r e s e n c eo f f a t t y a c i d s w i t h 3 a n d 4 d o u b l e b o n d s . ( 0 . 0 0 2 % i n e t h y l a l c o h o l )
;illl:
PHARMACODYN^MIC ^CTIVITY
3I9
c
o
g
c
L
ira
lave Lemth6n*;
F t o . l 2 l . S p e c t r a la n a l y s i so f t h e i n t e s t i n a lf a t t y a c i d so f r a t s a f t e r c h e m i c a l i s o m e r i z a t i o n s h o w s m i n i m a l a m o u n t so f m e m b e r sw i t h 3 . 4 o r m o r e d o u b l e b o n d s .( 0 . 0 0 2 V o
in ethyl alcohol)
.ffi{r',
ffii,j
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320
RESEARCH IN
PHYSIOPATHOLOCY
"l
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f a v e L e n g t h( n p )
Frc. 122. Spectral analysisof the fatty acids of the entire bo<ly of rats, after chemical isomerization shows the presenceof di- and triethenic members, and littlc of
m e m b e r sw i t h m o r e d o u b l e b o n d s . ( 0 . 0 0 2 V oi n e t h y l a l c o h o l )
ffi,-N
PH^RMAcoDYNAMTcAcrrvrTY
321
different lipoacidrcpreparationsby using a modificationof classicalmcthr.. (Note 3) Nlany mixtures of naturally occurring fafty
ods of conjugar..c
acids found in sapcnifiablefractionsu'cre conjugatedby the same method.
Conjugated di, tri, tetra, penta-and hexaenic fatty acids werc further
separatedfrom tl::sc itii:turesand studied,The unexpectcdrelationshipof
t h e s ec o n j u g a t c Jf , , l : ' , ' a c i d tso o x y g e nr v a so f i n t e r e s tW
. hilethe treatment
of linoleic acid at 37"C with oxygcn lcads ro progressiveincreasein the
amountof peroxidespresent,this doesnot occur for the conjugatedisomer
(FiS. 123) apparentlybecauseof repeateddestructionsof the peroxides
formed.
In general,the effectsproduced by conjugatedfatty acids at difterent
levels of the organizationare more intensiveand persistlonger than those
t2
(a)
to
cl
vf
de
z
o
8
e
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d
?o4
tr
,'
'-
--/t
,)
\---__-.,
,n,
\tv.,"-.
(b)
""
6
l 'i me ( hour s)
Frc. 123. Curves of peroxides of samples of linoleic acid (a) and its conjugated
isomer (b) induced tbrough the passageof oxygen ( 100 ml per minute for 30 cc of
sample) at 37'C. While peroxides are progressivelyincreasing i n t h e l i n o l e i c a c i d
prcparation,they do not change in the conjugated isomer,
322
n E S E , A R c Hr N p H y s r o p A T H o L o c y
PHARMACODYNAMIC ACTTVITY
//
323
324
xESEARcH
IN
PHYSToPATHoLoGY
P H A R M A C O D Y N ^ M I C ]^ I ' T I V I T Y
325
326
R E S E A R c Hr N P H Y S T o P A T H o L o c Y
PHARMACoDYNAMIC ACTtVTTY
327
and against which it would not be prepared to defend itself. This brought
us to the study of fatty acids having different nonpolar groups than those
of the normal and abnormal constituents.while most of these were prepared syntheticallyin the laboratory, we utilized on a substantiallylarge
scale two natural fatty acids which exist in plants and are sufficientlyheterogeneous,ricinoleic and crotonic acids.
Ricinoleic acid has a double bond between9 and l0 and a hydroxyl at
carbon 12 insteadof the seconddoublebond found in linoleicand linolenic
acids. As a result of the induction effect propagatedfrom the carboxyl
through the chain, the crr is a positive carbon. The positive characteris
enhancedby the adjaccntdouble bond betweence and c1e, and by the
hydroxyl bound to c12.C,, thusis very stronglypositive.we relatedthe intenselocal alkalosiswith consequent
watcr cxcretioncorresponding
to the
a l k a l i n ew a t e r yd i a r r h e at o t h c c f f e c to f r i c i n o l c i ca c i d l i b c r a t c di n t h e i n testine,and have consideredit to correspondto a local organicoffbalance
similar to that induccd in tissuesby unsaturatcdfatty acids.This would
explainthe intensivelaxativecflectof castoroil.
We uti.lizedricinoleic acid parenterallywith the aim of obtaining a
similar effcct in abnormalcellularand tissuelesions.The oily solutionof
ricinoleicacid has low toxicity when administeredparcnterally.However,
no manifesteffcct upon tumors or at differentlevelsof organizationwas
obtained.Crotonicacid did not showthe expectedinfluenceat theselevels.
Other hcterogeneous
agents,polyhydroxy fatty acids, were studied.
They were preparedby adding one or morc OH groupsto the nonpolar
groups at the double bonds of unsaturatedfatty acids.9, l0 dihydroxy
and 9, lO, 12, l3 tetrahydroxyfatty acidswere no differentthan thc corresponding unsaturatedfatty acids in their effects upon pain or systemic
analysesin humans,or upon tumor growth in animalsand humans.
Peroxide Fatty Acids
Peroxidesand epoxidesof fatty acidswcre preparedand studied.They
showedmore manifesteflectsupon virusesand bactcriain vitro and in vivo
than the other fatty acids. Investigationsof the effectsof these fatty acids
upon higher levels havc been limited until now. It seemsthat the effects
upon systemicand organic manifestationsare somewhatdifferent than
those obtained with use of the fatty acids from which the peroxidesand
epoxideswere derived. Influenceupon pain and upon tumors was grcater
for the correspondingunsaturatedfatty acids. This research---cspecially
with polyepoxidefatty acids-is still in progress.
328
R E S E A R C H
I N
P H Y S T O P A T H O L O G Y
t
!
c
o
L
h a y e L e n o t h( b N
F r c . 1 2 4 . I n f l u e n c ee x e r t e db y c h l o r i n e8 , a su p o n t h e c o n j u g a l e df a t t y a c i d s .A p a r a l l e l
d e c r e a s ei n t h e a m o u n t o f a l l t h e m e m b e r si s s e e n .( 0 . 0 0 2 % i n e t h y l a l c o h o l )
P H A R M A C o D Y N A M T c^ C r l v l r Y
3 2 9
330
nESEARcHrN pHysropATHoLocy
ence exerted by the carbonyl and methyl groups upon the intennediary
carbonsof the chain givesthe entire moleculean especiallyhigh reactivity.
This oppositeinfluenceis seenat its maximum in propanal,where cL suffers the influenceencrgeticauyoppositeof the carbonyl and methyl group.
The fact that, due to its relativelyhigh solubility in water, propanalwhich is a lipoid--{an be administeredin aqueoussolutionsand still act
upon the lipidic system-makes of it an especiallyinterestingagent.
we have investigatedthesegroups of lipoaldehydesfrom the point of
view of their influenceexertedupon the two offbalances.In the group with
unsaturatedshort chains, we studicd acrolein and crotonic and malcic
aldehydewithout seeingany specialeffect upon the other levels than the
cellular one, where a vacuolationwas obtained.Furthermore their toxicity
has represented
an handicap.More interestinghas appearcdthe group of
the saturatedshort moleculeswith odd number of carbons. while with
heptanalwe havc obtainedbesidesan influenceupon pain, also a manifest
inhibitoryeffect upon thc growth of expcrimentaltumors, it was propion
aldehydewhich has shown the most interestingcffectsupon pain.
This was secn for the group of aldehydeswith aliphatic saturated
chain such as propionit' and heptylic aldehl,tlesor with cyclic, as .rclicy'lic
aldehydes.Inadequatedoscs-f rom ,urocc. to 2 cc. of the l0% solutionof
propionicaldehyde,or of the I % solutionof the heptylicor salicylicaldchydes-a manifcstinlluencewas obtainedupon the systemiccondition as
well as upon pain. Patientsin offbalanceA with pain and generaldiscomfort, wcre seento have a decrease
of the symptomsaftcr the administration
of propionicaldehyde.The eflectupon tumorswas reducedand propionic
aldehydedid not changethe cvolutionof the tumorsin spiteof the marked
improvementof thc gencralconditionand even of the ccssationof pain.
Lipoids n'ith Thiol Groups
Mercaptans: According to thc systematizationof lipoids presented
above, a thiol group, acting as a polar group, will form a lipoid when
prepondcrantaliphaticor cyclic nonpolargroup.
bound to an cnergetically
In thc homologousrncrcaptanseries,evcn thc lowestmembersare lipoids
forcesof the thiol group.
becauscof the weak electrostatic
Although methyl mercaptanis a lipoid accordingto our classification,
this substanceis too volatilc to be used.Therefore,the first low membcr
was ethyl mercaptan.
of this homologoussericsto be investigated
The effectsof ethyl mcrcaptanupon microbeswere more limited than
those seen for fatty acids. To determinethe effectsat the different levels
PHAR,M^CODYN^MIC ACTIVITY
331
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782
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77,!8
776
7.7a
712
dd"
+
t
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7.62
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F l c . 1 2 5 . S e c o n dd a y w o u n d c r u s t p H s h o w sa c o n s t a n tc h a n g et o w a r d m o r e a l k a l i n c
v a l u e s f o r a l l t h e s u b s t a n c e sh a v i n g s u l f u r i n t h e i r p o l a r g r o u p .
332
nESEARcHIN PHYStoPATHoLocY
AC T I V I TY
1 1 1
-t .1 .1
creascd.For examplc.cflectswerc considerablyrcducedfor hexyl mcrcilptan as comparedwith ethyl mercaptan.Thc secondgroup-thc heptyl and
'I
allyl mercaptans-procJucedmorc intensivceffccts. his was espcciallytrue
for allyl mercaptan.Membersof the third group-rvith longercarbon chains
such as dodecyland hexadccylmcrcaptans-producedcflccts so slight that
ttrey seemedalmost nonexistent,exceptupon pH of the secondday wountl
crust, which showed valucs far abovc the rangc of thc controls, just as
with other membersof this homologousscrics.
Yesks
Control
ooo
Start treatoent
Treated
rl
ethyl
ercaptan
Treatedrith
ethyl
mcaptan
OOOI
Start treatnenl
U l s e r arto n
e O - .
i;.
F t c . l l 6 . l n f l u c n c e c x c r t e d h 1 ' c t h r ' l n r c r c a p t i u l u p ( ) n l \ : r r c o n l i r r n d t r e c t lb l , h e n z pyrene. For biggcr lumor\ jt induccs constltntly ln ulccration, while for rmall tun)ori,
thcir disappearance.
334
RESEARcHrN pHysropATHoLocy
tions that appearedto have a bivalent sulfur bond at the nonpolar group.
We investigatedcolloidal sulfur which, if introduced in the organism,
seemedto undergo changessimrlar to those of bivalent sulfur. We found,
thus, that the sulfur absorbedaftcr being administcredin colloidal form,
was almost entirely eliminatedafter oxidation in the form o[ sulfates.In
animals,no pharmacodynamicinfluenceswere observed.OnIy the pH of
the secondday wound crust was increasedwhen thesepreparationswcre
administeredparenterally in suspensionor were given orally mixed rvith
food, in proportions up to 4%. There was no evident change in tumor
evolution in animalsor humans.
Hydro-Persulfides
Another sulfur compound,so-called"sulfurizedoil," in which sulfur
and fatty acidsappearto form a hydropersulfide,
(Note 7) was tested.This
hydropersulfidepreparation,althoughit exhibitedno influenceupon viruses
in vitro, did inducea good degreeof local resistanceof the skin to smallpox
infection.The effectsupon microbeswere reduced.There was little direct
influenceexerted upon cells. The preparationswith 0.5 to lVo srtfur
bound to cottonseedoil were well toleratedlocallv when administeredintramuscularlyor intraperitoneally.
The effect of parentera.land oral hydropcrsulfrdeupon pain was slow
to appear,in contrast to the effect of fatty acids and even of mcrcaptans.
However, it persistedfor a long time. Pain of the acid pattern was eased;
pain of alkaline pattern was exacerbated.The influenceupon the second
day wound crust pH was marked.The local pH increasedto valueseven
higherthan 7.80. In radiationlesions,the dimensionof ulcerationincreased
and healingwas retardedor even prevented.In some tumors in animals,
the rate of groMh was slorved.This latter effect was not uniform in the
different types of tumors tested and even in the same type of tumor in
different groups o[ animals. systemic changesalso varied. Doses correspondingto 5 mgr. of sulfur were not toxic for 30 grn. mice in a single injection.Nor were l5 mgr. dosesin 200 gr. rats. chronic toxicity studies
showedthat 0.2 mgr. daily injectionsin mice and 5 mgr. injectionsin rats
for as long as three monthsdid not inducepathologicalchanges.High doses
suchas I to 2 cc. of a lVo preparationinjectedseveraltimes a day in humanswas almost uniformly followed by a rise in temperature,usually after
a few davs.
PH^RM^CODYN^MIC
ACTIVTTY
335
336
p . E S E A R c Hr N p H y s t o p A T H o L o c y
PH^RMACODYNAMTC^CTlvtTY
33'1
338
xESEARcH IN pHYSropATHoLocy
PH^RM^CODYN^MtC
^CTIVITY
339
340
nEsEARcH rN pHysropATHoLocy
PHARMACODYNAMIC
ACTtvrrY
341
342
xESEARcH rN pHysropATHoLocy
Selenium Lipoids
The influence exerted by bivalent sulfur upon oxidation processesin
which fatty acids participate,servedas a guide for further research.Seeking agentsthat would act at a still lower levcl of the organization,even
below the cclls,we consideredother substances
which also affectoxidation
processes.Theoretically,at least, it appearedpossibleto induce changes
at a compartmentbelow the metazoic,at which sulfhydryl-containingcompoundsact.
We have discussedpreviouslythe systematization
of the biologicalactivity of elements,their fundamentalanti-A and anti-D influence,their distribution among the various levels of the organism, all related to their
atomic structureand their place in the periodic chart.
All of this led us to investigateseleniumwhich is the nonmetal element next to sulfur in the sixth seriesof Mendeleeffsperiodic table-the
serieswith an anti-A characterin which oxygenis the first member.According to its period, seleniumbelongsto the cellular compartment.
The first problem was the nature of the compound in which it would
be active.We wcre particularly interestedin using bivalent negativeselenium becauseof the activity of bivalent negativesulfur. However, we did
investigatethe selenicand seleniousacids.Theseacidsor their sodium salts
PHARMACODYN^MIC
^CTIVITY
343
344
RESEARcHIN PHYSIoPATHoLocY
PHARMAcoDYNAMTC
ACrrvtTY
345
ALKYLATING
AGENTS
We investigated,as compounds with negative character, certain alkylating agents,choosing from the large number availablethose which also
showed lipidic properties.We were especiallyinterestedin two members,
sulfur mustardand epichlorohydrine.
Sulfur mustardcontains,along with
one active polar chloroethyl group, a secondrepresentedby the bivalent
sulfur polar group. It has the effect at different levels of organizationof
producing an offbalancewith predominanceof the acid lipids. We will
discussbriefly here some of the experimentsin which this influenceupon
the body lipids has been observed.
346 /
x E s E A R c Hr N P H Y s r o P A T H o L o o y
Su$ur Mustard
In studying sulfur mustard,we were first interestedin its effectsupon
body lipids and, through them, upon the lipidic systemof organisms.In inducing sulfur mustard'scharacteristicskin lesions,an interestingrelationship wasobserved.Pure sulfur mustardwas appliedto mechanicallyepilated
skin of rats. If a suffficientamount-2 to 3 drops-was used and spread on
one squarecm., the animal died. However,the time of death varied. If the
lesionshoweda massivenecrosis,followed by deep ulceration,similar to a
burn of the third degree,the animal died in about three weeks. However,
if the lesion was only erythematous,similar to a burn of the first degree,
the animal died in only 3 to 4 days. It seemedas if the lesion itself intervened secondarilyin the pathogenesisof changesleading to death. Sick
but still living cells appearto have an activity which is highly detrimental.
The abnormal cells apparentlyproduce substanceswhich are responsiblefor
rapid death of the animal. In widely necrotic tissues,these abnormal but
still living cells are limited in number; in an erythematous lesion, they
form the lesion itself. This correlation of toxicity with local Iesion was confirmed by the fact that excision of the lesion itself, il performed in time,
preventeddeath in some animals.The administrationof ferrous sulfate to
rats having sulfur mustard applied to their skin was seen to induce the
erythematousform of the lesions,with death in 3-4 days.
The similarity between the influence exerted by mustard burns and
caloric burns with a sufficientlyextensivefirst degreeburn producing more
rapid death than a third degreeburn, was of interest.
Analysis of the body of an animsl killed by a mustard burn reveals
abnormal amounts of unsaturatedfatty acids and reduced amounts of
sterols. In some cases,where death occurred after more than three weeks,
body sterols were found to be almost completelylacking. In these cases,
almost no insaponifiablefraction could be found. The lesion itself, especially an erythematousand edematousone, was very rich in unsaturated
fatty acids.Histologicalstudy of theseskin lesionsrevealedchangessimilar
to those obtained through the intradermic injection of concentratedsolutions of body acid lipidic fractions.The study of these lesionsfurther revealedthat the lesionsthemselveswere separatedfrom the organism by a
barrier of adipouscells,the resultof an exaggerationin number of the cells
of the subdermicfatty layer.
We studied these important changesfrom severalpoints of view. We
could show that an exaggerationof the adipouslayer underneaththe skin
occurs wben lipoids with negativecharacter,such as polyunsaturatedfatty
PHARMACODYNAMTC ACTMTY
347
348
xESEARcH rN pHysropATHoLocy
Epichlorohydrine
The importance of the relationship betweenthe energeticcenters present in alkylating agentsand their ability to produce type D offbalance, led
us to study a group of these substanceswhich, at once, have both short
molecules and two polar groups in close proximity. The desire to have
such an agent with lipoidic property as well led us to study epichlorohydrine which correspondsto propane and has an epoxy group binding C2
and C3, while cr binds a chlorine. Solublein neutral solvents,epichlorohydrine becomes soluble in water only after hydrolysis. Its biological
activity differs from other chlorohydrinessuch as chloropropanediolor
trichloropropane,both of which can be consideredto be closely related to
the substanceproducedby hydrolysisof epichlorohydrine.The acute toxic
dose of epichlorohydrinewas found to be 6 mgr./30 gr. for mice and 25
mgr./100gr. for rats by intraperitoneal
administration;22and 35 mg./100
for mice and rats by subcutaneous
injection.In testsfor chronic toxicity, it
was apparentthat dosesof 5 and L5 mgr. injecteddaily were well tolerated
respectivelyby rats and mice. with higherdoses,the animalsbecamerapidly
emaciatedbeforedying. Used orally in drinking warer,a solutionof l/3000
was well toleratedby rats and mice even for months.With the use of solutions of l/2000, only a few animalsdid not lose weight,while a solution of
l/1000 invariablyinducedwcight loss.
There were no effectsobservedupon microbesor bacteriophage.
It appearsthat epichlorohydrine,actingbelow the morphologicallevels,
induceschangessimilar to thoseseenfor other alkylating agents.However,
it is not upon the desoxyribo-nucleic
acidspresentthat an important action
is seen but in the lipidic system at these lower levels. Epichlorohydrine
seemsto act also at other levels.The influenceupon pain-an increasefor
alkaline pattern, a decreasefor acid-was more noticeablethan for sulfur
compounds.Delaycd eftects,however,were more obvious than immediate
ones. The influence upon wound healing was similar to that of polyunsaturatedfatty acids. Cancerouscells, such as those from mouse ascites,
were destroyedin vitro by a 0.5 solutionof epichlorohydrine.The effect in
vivo upon sarcoma 180 or Ehrlich ascitestumors was most interesting.
Administeredby subcutaneous
or intramuscularinjection,epichlorohydrine
had no effecton the tumor even in dosesas high as 2,5 mgr. daily. However,
when administeredin drinking water in a 1/1000 solution,it prevented
the developmentof ascitesin 19/20 animals.But the toxicity was too high.
A L/2000 solution,usedas drinking water,controlledthe condition in morc
than 50o/aof the animals, while a I /3000 solution showed favorable re-
P H A R M A C o D Y N A M T CA C ' r l v t r Y
349
350
xEsE^RcH rN pHysropATHoLocy
PHARM^CODYNAMTC ACTTvTTY
351
Amount of Element
at the Higher Level
high
Iow
high
high
high
low
low
low
Interpretation
: Occurrence
at the Proper Level
quantitativeexcess
qualitativeinsufficiency
qualitativeexcessive
utilization
quantitativedeficiency
352
xEsEARcH tN pHystop^THoLocy
trr
Excess
E
+
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(t)
TotalBloodK+rtq
F r c . 1 2 7 . T h e r e l a t j o n s h i pb e t w e e nt h e a m o u n t o f p o t a s s i u mi n s e r u m a n d i n t o t a l
b l o o d p e r m i t s t o i n d i c a t et h e e x i s t i n gc o n d i t i o n a s b e i n g i n n o r m a l l i m i t s , i n q u a n r i tative deficiencyor excess,or in an offbalancetype A or D.
PHARMACODYNAMIC ACTIVITY
,,/ 353
Metals
Be
Mg
Ca Sc V
S r Y Nb
Ta
Ba
Ce N d
Ra Th U
Non-Metals
c
si
Mn
Tc
Re
Sm
Pu
Co C u
Rh Ag
Ir
Au
C d Dy
Cm Cf
Ge
Sn
Pb
Er
o
s
Se
Te
Yb
354
REsEARcH rN pHysropATHoLocy
P H A R M A C O D Y N ^ M I CA C T I V I T Y
,/
355
356
R E s E A R C Hl N
pHystopATHoLocy
P H A R M ^ C o D Y N A M t C A C r l v r T Y
3 5 7
358
xESEARcH rN pHysropATHoLocy
G
($
F r c . 1 2 8 . T h e i n f l u e n c ee x e r t e d b y t w o e l e m e n t s ,p o t a s s i u ma n d c a l c i u m , u p o n t h e
second day wound crust pH shows a frank tendency toward acidification for potassium and alkalinization for calcium. For diflerent salts, the differences result from
t h e u n e q u a l i n f l u e n c ee x e r t e d b y t h e a n i o n w h i c h w o r k s a d d i t i v e t y t o t h a t o f t h e
cation.
358
r.ESEARcHrN pHysropATHoLocy
r L
0 v
LL)
I
|
o
E-o
f . i L
Oo
G
|.!
F r c , 1 2 8 . T h e i n f l u e n c ec x c r t e d b y t w o e l e m e n t s ,p o t a s s i u ma n d c a l c i u m , u p o n t h e
second day wound crust pH shows a frank tendency toward acidification for potassium and alkalinization for calcium. For diflerent salts, the differences result from
t h e u n e q u a l i n f l u e n c ee x e r t e d b y t h e a n i o n w h i c h w o r k s a d d i t i v e l y t o t h a t o f t h e
cation.
PH^RM^CODYNAMIC ACTIVITY
359
360
nEsEARcH rN pHystopATHoLocy
r
.i::
-ia$^
, :, ;
. a:ia ii,*:.tj1l\!.rl
n:!ii:
iil\r*#iiini
a
I
P H A R M A C O D YN ^ M I C
A C T I V I T Y
361
Heavier Elements
we have studiedthe activity of elementscorrespondingto the lower
lgvsl5-5ugh as strontium and tin for the nuclear;barium, gold and lead
for the submorphologic;and cesium for rhe primary biological compartments. The lower the level of the element,the greater appearsto be the
preventiveeftect againstinduction o[ tumors by carcinogens.But the minimal, or completeabsenceof effect upon alreadyexistingtumor cells remains unchanged.
As mentionedabove,we conncctedthis paradoxicalactivityto the fact
that, while normalcellsare able to manufacturecompoundsthroughwhich
the appearanceof a cancerousentity can be prcvented,these compounds
are no longerformed if an cntity is alreadycancerous.
These have been the considerations
forming the basis for an entire
seriesof studiesof the role of variouselementsin the pathogenesis
of
cancer.
Elements in the Pathogene,sis
ol Canc'er
Investigationshave been made of the amount and form of the element presentin the normal animal as contparedto the cancerousanimal.
The quantitativeand especiallyqualitativedifferenceshave been seen to
indicatethe site of the abnormalitylargelyresponsible
for the lack of influenceexertedby the element.The resultsexplainwhy the administration
of the elementalone is unable to influencethe cvolution of an a-lready
existingcancerousprocess.More interesting,
they show what compoundof
the elementcould have an influence.This research,which is in progress,
opens the door for possibletherapeuticapplications.It is through such
compounds,presentin the normal and lackingin the cancer-stricken
animal, that attemptsare beingmadeto influencethe evolutionof cancer.An
important step has been the finding that suitablecompoundscan be obtained by the treatmentof fresh organsin vitro with some of the elements.
Their study may make possiblesyntheticpreparationof suitable compounds.The few resultsalreadyobtainedin experiments
with animalsconfirm that D-inducing activity representsa factor which the promising elementsshare.
C H A P T EI R5
PHARMACODYNAMIC
ACTTVTTY(PART TW O)
ANTI.FATTY ACID GROUP
ID
ll- nnellrL
To INVEsTTcATIoN
oF AcENTScapable of correcting offbal-
p H A R M A C o D y N A M T cA c r t v t r y
(p,rnr rwo)
363
one, a relatively
wereinvestigatcd:
In this study,two typesof influences
direct effect induced through a neutralizationof the energeticcenters resulting in a more or less advanceddegreeof inactivationof the fatty acid;
the second,an indirect effect achievedthrough changcsin the metabolic
processes
in which fatty acidsintervene,In a differentkind of intervention,
the anti-fatty acid to which a fatty acid is bound governs its ultimate
biological fate. For example,the bond to glycerol favors caloric metabolism. The bond to glycerophosphoricion converts a fatty acid, saturated
as well as unsaturated,into an organizationalconstituent.The bond to
sterolsfavors a functional role, even for monoethenoids.
We started the study of the naturally occurring anti-fatty acids with
those agents known to be bound to fatty acids in the organism. The
simplestsuch agentis glycerol.
Glycerol
Glycerolis the most ubiquitousfatty acid-bindingsubstancein nature.
We attempt to explain this fact on the basisof glycerol'sstructureand the
specialbiological role it confers.We have seen that fatty acid molecules
take reciprocal parallel positions when they form monomolecularlayers.
In their bond to glycerol,thesefatty acid moleculesconservethis recipro(Fig. 129)Thiscould cxplainwhy, in the body, the bond of
cal relationship.
being
fatty acid and glycerolalwaysis a triglyceride,mono and digJycerides
(?)
CD
(-)
c)
(-)
(J
O
O
c(\.1) o
- t -: E
a
(?)
cD
(J
(.)
co
(J
cf)
CJ
cf)
(J
(7)
(-)
Io
C\J
-(J
F t c . 1 2 9 . T h e f a t t y a c i d r a d i c a l st a k e i n t h e t r i g l y c c r i d em o l e c u l e sa s i m i l a r p a r a l l e l
position as when they form a monomolecular la1'erat the surface of water.
364
nESEARcHrN PHYsropATHoLocY
only intermediatesteps.The samefatty acid has different biological activities if administeredas free acid or as triglyceride.The combinationseems
to serveas an energy-furnishingmetabolite.Bound to glycerol,fatty acids
with long or short chains, saturated,monounsaturated,polyunsaturated,
and even conjugated,seem to representenergeticreserveswhich are utilized as caloric metabolites,especiallyin those specieswhich are able to
storethem.
With this relationshipin mind. we administeredglycerolwith two objectives: I ) to obtain, as an immediateeffect,the inactivationof the free
fatty acids presentin abnormalconditionsthrough the neutralizationof
their polar groups, and 2) to eliminatethesefatty acids by turning them
into caloric metabolites.
Studying the activity of glycerol at different levels, we could see no
influence upon phages. However, an indirect effect was observed upon
viruses.Glycerol is widely used as a specialmedium for the preservation
of virusesin tissucs.Its preservative
value can be correlated,at least in
part, with its influenceupon fatty acids.We have seenthat fatty acids have
a noxious effect upon viruses,leading to their disappearancein various
organs. The treatment with purc glycerol reduces autolysis of organs
through a dehydrationeffect.Curtailingthe lytic activity of the enrymes
active in autolysis,glycerol reduccsthe amount of fatty acids liberated
through such autolysis,and thus preventsthe destructionof the virus.
Glycerolmay also preservevirusesby acting antagonistically
to any fatty
acids which still manageto appear.
Glyccrol has a bacteriostaticeffectupon only a few speciesof microbes
and only when appliedin high concentrations.
A minimal influenceupon cells was seenfor glycerolin Tetrahymena
pyriformis and ascitestumor cells. To study its action at higher levels,
glycerol was administeredorally or parenterallyto animals or humans.
Solutionsof ZOVoglycerol were well tolerated when injected subcutaneously or intramuscularly.It should be noted that when glycerol was administeredto complcx organisms,it was largely absorbedand circulated
without alteration,a fact which would explain the effectiveness
of relatively small amounts.At the tissuelevel.glycerolinduceda changeof the
local pH of a lesiontoward the acid side,as seenin the secondday wound
crust. Figure 130 illustratesthis. The changeexplainsglycerol'saction in
increasingintensityof acid pattern pain and decreasingintensityof the
alkaline.This influenceupon pain was obtainedconstantlywith very small
amounts,permittingthe use of glycerolevenas a testfor diagnosisof pain
pattern.Intramuscularinjectionof t/z cc. of a 2Q% solutionor oral ad-
p H A R M A c o D y N A MAt c r r v r r y ( p , r n r r w o )
365
minisuation of. Vz cc. of a 5oo/osolution in water has been used for this
purpose. However, later, when other agcntswere found to produce even
more overt responses,we stoppedusing gJycerolas a routine test.
Glycerol has almost no beneficialinfluenceupon the healingof wounds
or radiationlesions.Healingwas evenretardedin someexperiments.
Various changesin the evolution of tumors occur when host or transplant are
trcated with g.lycerol.In some, these changesare minimal; in others an
obvious reduction in grofih occurs. In a high proportion (12/20), a
marked involution has been noted for Walker tumors in rats. Ascites Sa
180 in mice, after repcatedintraperitoneal
injectionsof a solution of.ZVo
GI ycerol
F r c . l l 0 . C l y c e r o l i n d u c e sa l o w e r i n go f t h e s e c o n dd a y
wound crust pH.
aoaa
aa
.O
366 /
RESEARCH IN
PHYSIOPATTIOLOOY
OO
Control
Start t reatnent
Ireatedrith
Glycero
I
rith
Treated
Glycerol
Ireatedrith
Glycerol
. l
. l
. 1
I -,.
pHARMAcoDyNAMtc
(p,rnr rwo)
^crtvrry
,/
367
E , *rr
?1
2{f,2e
2
27 to5//l
- J T J J J J I J I J J I ,
S p .l6
Grav.
, 6o
Z8
tu
62
pHf.1
Cll.a
/
S . T61.
E
J
L
(u
v,
E
+
><
F r c . 1 3 2 . T h e a d m i n i s t r a t i o no f I c c g l y c e r o l d a i l y i n d u c e sa c h a n g eo f t h e u r i n a r y
pH toward thc type A, before changesin the other analysestake place.
368
REsEARcHrN pHysropATHoLocy
p H A R M A c o D y N A M T c^ c r t v t r y
( plnr
rwo)
369
370
nESEARcH rN PHYSToPATHoLocY
p H A R M A c o D y N A M I cA C r t v r T y ( n r n r
rwo)
371
372
nEsEARcH rN PHYSropATHoLocy
p H A R M A c o D y N ^ M l c A c T - r v r T y( p , r n r r w o )
373
374
x E S E A R C I {r N p H y s r o p A r H o L o c y
donicacid.About 25Vo of the total fatty acid contentof the glandsis made
up of this acid which is found only in smallquantitiesin other organs.We
alsofound pentaencs
and hcxaenes
prcscntin greateramountsthan in other
organs.These fatty acids nrust have a biological purpose and two hypothesescan be advanced.Accordingto one mcntionedpreviously,(Note I I,
Chapter VI), the corticoidswould be synthesized
from arachidonicacid
throughcyclization.Accordingto thc sccondhypothesis,arachidonicacid,
as well as other high fatty acidsprcsentin the adrenals,would be used as
activefunctionalfatty acids.Secreted
by the adrenals,they would passinto
the circulation and intcrvene as nccded by the organism, especiallyfor
immediatedefcnsepurposcs.
We have seenthat an interventionof polyunsaturatedfatty acids occurs
in the first defcnsercsponscof an organismto a noxious agent.These acids
for the exaggeratcd
are responsible
through which the
oxidationprocesses
organismattacksthe noxiousagentsthemselves
or the heterogenized
constituentsresultingfrom their action. Wc considerthat some of the fatty
acidsinterveningin this defensemcchanismare liberatedlocally,especially
if they appearin responscto a conditionlimited to a lower entity. In this
case,they would come from changesinduccd in the constituentsof the
entity itself. Thc local interventionof lipolytic enzymeswould lead to a
liberationof free fatty acids.In a defcnseresponsefor the organismas the
highestentity, thc activcly intervcningfatty acids appear in the general
circulationin the first phaseof the diphasicphenomenon.Some of these
fatty acids would be of adrcnal origin, liberatedat these moments.In the
secondphaseof thc defensemechanism,
a further liberationof steroidsby
the adrenalswould occur, aimed at counteractingthe effectsof fatty acids.
The diphasicsystemicprocesswhich rcsultscan be consideredto represent
which occur alternatelyand which, through
an exaggcrationof the proccsses
normal oscillations,insurethe dynamic systemicba.lance.
The adrenalsconccivablycontrol abnormalfatty acid activityby their
quantitativelyexaggcrated
interventionand by releaseof qualitativelyabnormal products that would pass into the circulation and result in offbalances.The activity of the adrcnirlsin counteractingthe influenceexercisedby fatty acids has been made the subjectof a specialinvestigation
in our laboratoriesby E. F. Taskier.
By comparingthe doscsof an agentrcquiredto kill normal and adrenalectomizedanimals, it has been possiblenot only to identify this intervention but to judge thc degreeof this specificdefensemechanism.The
"Adrenal DefenseIndex" for an agent-the ratio betweenthe minimal
animals represents
lethal dose in normal animalsand in adrenalectomized
p H A R M ^ c o D y N A MA
r cc r r v r r y ( r l n r
rwo)
375
tY"1,:'
i,*:',i
jgf;r-J{.9!1:;'.';.r
i}.\-'ifl..il'::r.:;;
?5sJJf,;.ftrtalfg'.?aG,E-*r
if
'-S
l'*
, i ;:16',"1'";
l?6
RDSIJA RCI{
I N
PI"IYSIOPA']'}IOLOGY
t{gc-9-ry
H. Butyl
Alcohol
S e c .B u t y l TertiaryButyl
Alcohol
AIcohoI
Q rch
\IH
I sobutyl
Alcohol
F l c . 1 3 3 . S c h e m r r t i cr c p r e s c n t a t i o no f t h e n r o l e c u l l r s u r f a c e so f t h e . l i s o m c r s t r f
b u t a n o l . T h e c o n s t a n t b o f t h c v a n t l e r W a a l s f o r c e s r c l a t c d t o t h e i r s u r f a c e sa r e
u n e q u a l .A n l i n i m u n r v a l t r c i s s e c n f o r a l n r o s ts p h e r i c n r o l c c u l eo f l c r t i a r y b u t a n o l ,
a f a c t w h i c h e x p l a i n st h e n o n p r c d o m i n a n c co f t h e p o l a r g r o u p i n t h i s m o l e c u l e .r e s p e c t i v e l yi t s n o n - l i p o i d i cc h a r a c t c r .
o
g
(tt
=
co
I
c
o
g
(It
ct
I
(J
OJ
v,
m
I
(f,
.Fl
c)
c
rO
3
m
I
o,
p H A R . M A c o D y N A M t cA c r r v t r y
(p,lnr .rwo)
3jj
n - B u t y l s e c - t s u t tyel n t - B u t ylls o b u t y r
F r c . 1 3 5 . W h i l e t h e 3 i s o m e r so f b u t a n o lw h i c h a r e l i p o i d si n f l u e n c et h e s e c o n dd a y
w o u n dc r u s t p H . l o w e r i n gi t s v a l u e s .t c r t i a r y b u t l n o l w h i c h i s n o t a l i p o i d , d o e s n o t
i n f l u e n c ei t .
378
RESE^RcH tN pHysropATHoLocy
29o
Contro I s
Butanol
Agein reeks
F t c . 1 3 6 . E . f f e cot f 0 . 5 9 ' bs o l u t i o n o f .n - b u ! a n o la d m i n i s t e r e di n s t e a do f d r i n k i n g w a t e r
u p o n t h c i n c r e a s ei n w e i g h t o f y o u n g r a t s . T h e v a l u c s r e p r c s e n tt h e a v e r a g ef o r 2 0
f e m a l e s ( . . . . ) . N o d i f f e r e n c e sa r e s e e n f r o m n o n t r e a t e dc o n t r o l s ( - ) .
p H A R M A C o D y N A M T cA c r t v r r y
(p,rnr rwo)
3'79
Controls
Butanol
/6
20
24
29
J2
Ageln t|eeks
F r c . 1 3 7 .T h e s a m c d a i l y c h a n g es c e ni n m a l e r a t s
380
./
RESEARCH IN
pHystopATltoLocy
p H A R M A C o D y N A M t cA c r r v r T y ( p , r n r r w o )
3gl
loo
-No Loctole
./
90
, Untreofed
80
-No
70
=
o
Cl
LI,
. / n - B u fo n o l
' .. t/
NoCl
50
ae
30
n- Bulonol
No loclole
' f r
t/,'
20
i / ''
ro
,'
/I,,,,
6
T i m e ( h r s ).
F t c . 1 3 8 . I n f l u e n c ee x e r t e d b y d i f f e r e n t a g e n t su p o n t h e m o r t a l i t y o f m i c e s c a l d e d
f o r 3 s e c o n d si n w a t e r a t 9 0 ' C . W h i l e s o d i u m l a c t a t e s e e m se v e D t o i n c r e a s et h e
m o r t a l i t y a n d N a C l a n d b u t a n o l i n s a l i n eh a v e l i t t l e i n f l u c n c e .a m a r k e d p r o l o n g a t i o n
o f t h e s u r v i v a l t i m e i s i n d u c e db y t h e m i x t u r e o f n - b u t a n o l a n d N a l a c t a t e .
ffi
::"v:''i:*a:.1?riaL:
*i.
;r:'rii:
*,
?'
.i J, .,' -; j ;,:):i I-r\1,:;-*
U0irir{., ;'--fj'",&'}.n':'.i;.q:
hqf*,SJt..'',{r
i:.:_*{fjlt
gj :r,f
382
p . E s E ^ R c l {r N p H y s r o p A T H o L o c y
92
n-HeptyI
n-Octyl
n-l,lonyl
778
776
PH
oo
o
o
771
772
o-o
o
o
oo
7
ooo
1
o
o
oooo
oo
o
:6
F t c . 1 3 9 . T h e i n f l u e n c ee x e r t e db y h e p t a n o l ,o c t a n o la n d n o n a n o l u p o n t h e s . d . c .p H .
W h i l e h e p t a n o l i n d u c e s c o n s t a n t l ya l o w e r i n g o f t h e a l k a l i n i t y o f t h e s e c o n d d a 1 ,
w o u n d c r u s t , t h i s c f f e c t i s l c s sc o n s t a n tf o r o c t a n o la n d n i l f o r n o n a n o l .
p H A R M A C O D y N A M T CA C r r v r T y ( p ^ R T T w o )
393
384
,/
sESEARcHtN pHysropATHoLocy
/0
rl
tt
tS
H e p t a In-oB u t a nI o
Sp.
trn
rav.
I
,.
)
).
)
nl-{, 6
l./r
6
6.
6,2
6.o
J,E
a6
5.,/
68
<A
6,t
7
.1.6e
6c
tt
J(
) t
a 1
6 )
E c i
l 6 l
L a /
$ t.t
alrs7
J:'
cr!
.A
+
)z
3:./
4. 1
4:7
t(l
y a
J{
l)
F l c . 1 4 0 . l n a p a t i e n tw i t h b o n e m e t a s t a t i cl e s . i o n tsh e a d m i n i s t r a t i o no f h e p t a n o l a n d
b u t a n o l s h o w s a p r o g r e s s i v ed c c r e a s ei n t h e v a l u e so f s e r u m p o t a s s i u ma n d i n c r e a s e
i n t h e u r i n a r y p H , t o w a r d o f f b a l a n c et y p e A .
p H A R M A c o D y N A M TAc c r r v r r y ( p e n r r w o )
385
o
o
to
lr3
/
/
ti
/
r----t/ " ,
Or
i
5
20
.02
.o4
06
.o8
.to
.t2
,t4
.t6
M o l o r C o n c e n l r o l t o n- E u l o n o l
F r c . l 4 l . C l o t r e t r a c t i o n ,m e a s u r e da s p e r c e n tw e i g h t o f c l o t / t o t a l weight of blood,
plotted against molar concentration of butanol in blood. Different symbols used for
blood samplcs of cach animal.
386
RESEARCH IN
PHYSIOPATHOLOGY
R
I
60
o
o
I
02
04
06
Molor
08
Concenlrolton
to
t?
t4
.t6
- Bulonol
Frc. 142. Clot retraction. measuredas percent weight of clot/total weiBht of blood,
p l o t t e d a g a i n s ts i m i l a r c o n c e n t r a t i o no f b u t a n o l i n b l o o d . A v e r a g e so f d i f f e r e n t a g e
g r o u p a n i m a l sw e r e s t u d i e d .
Jc""*J.$J
\
l"
as
\e 80
Eoo
't!
I
'20
04
08
t2
t6
20
.24
}--m
32
.48
p H A R M A c o D y N A M tA
ccrlvtry (p,rnrrwo)
387
Polyols
In another study, we consideredthe polyalcohols,bearing in mind the
important role playcd by glycerol in the biological acrivity of lipids. In
animals,ethylene-glycol
and diethylene-glycol
proved to be too toxic for
parenteraladministration.However, near toxic dosesproduced interesting
resultsespeciallyin tumors. Even in relativelysmall subcutaneousWalker
rat tumors, 2.5 cm. in diameter,for instance,necrosiswas constantlyinduced and followed by skin ulceration. The characteristicinfluence of
these alcohols was to induce a necrotic processnot limited to the tumor
alone but affectingsurroundingtissues,
1.2 Diols
By following the influenceexertedby more than one hydroxyl in the
molecule,we tried to relate propertiesof glycerol to those of aliphatic
lipoids.we preparedlipoids havinga polar formarionof 2 or 3 hydroxyls
bound to the first carbonsof an aliphaticchain. The lipoidic charactcr
was inducedby the lengthof the chain. To preparethcsesubstances,
we
started with correspondingalph-hydroxyfatty acids in which the carboxyl
was reduced to a primary alcohol by treatmentwith lithium aluminum
hydride.As a prototype,we studied1.2 octanediol.The lipoidic character
was recognizedby its high solubilityin neutralsolventsand a limited solubility in water.
There were no marked diflerencesbetweenthe effcctsof octanol and
1.2 octanediolin systemicanalyses.Both raisedsurfacetensionvaluesin
particular.However, the new componenthad an effect upon the central
nervoussystemdifferentfrom most other higher alcohols.As mentioned
above, thesealiphaticmono-alcoholsdo not induce convulsionswithout
the concomitantinterventionof anotherfactor. The secondfactor can be
a local condition in the nervoussystemitself, as in subjcctswith cerebral
tumors,or otherswho have had previousconvulsions.It can also be another substance;desoxycorticosterol,
coraminc,glycerolor glucose,when
administered
with octanol,for example,inducedconyulsionsin some subjects.However,1.2 octanediol,in rcpeatcddosesof around200 mgr. daily,
inducedconvulsionsby itself.This could be explainedby the facr that 1.2
octanediolcontainsin its moleculca group which energeticallyresemblcs
glycerol.Beyond this effect,thcre were no manifestdifferencesbetween
this substanceand correspondingmono-alcoholsin influenceupon pain,
tumor growth or systemicmanifestations.
388
x E S E ^ R C Ht N p H y s r o p A T H o L o c y
pHARM^coDyN^Mr( ^crrvrry
(p,r*r rwo)
lgg
390
/'
RESEARCH IN
PHYSIoPATHoLOOY
Lipamines
Theoretically,it was to be expectedthat lipids with an amine as polar
group would have a marked anti-fatty acid action. We studied severalof
these substancesfrom the standpoint of their influence upon physiopathologicalchangesconsideredrelatedto lipoidic predominance.The first lipoid
of the aliphatic amine seriesis hexylamine.A nonpolar group of at least
6 carbons is required for predominanceover the potent amino radical.
From commercial sources,we obtained aminescorrespondingto the usual
saturatedfatty acids with cven numbersof carbons,ranging from 6 to l8;
a few unsaturatedwith 18 carbons;and heptylaminewith an odd number
of carbons.All thesecompounds,when injected in mice and rats produced
severelocal reactions,often followed by skin ulcerations,even when administeredin oily solutions.For this reason,we tried salts of theseamines
usuallyobtainedwith aceticor hydrochloricacid. Saltsof the lower members of the series,no longer had lipoidic character.However, we could use
hexylaminein an oily solution for intramuscularinjection. It appearedto
be rclatively well tolerated locally evcn in humans. No apparent changes
were seen, however, in systemicanalyses,and the immediate and long
range effects upon pain were minimal. No changes were obtained in experimental tumors except by local treatment,as in ascitestumor, or by
injectingthe product at the level of the transplantitself, in which case the
growth of the tumor was slowedor even halted. A similar effect was seen
when the transplantwas dipped in the oily solution of the product and the
procedurewas repcatedin successive
generations.
On a larger scale,we utilized, both in animals and humans. the salt
obtained from hexylaminewith nicotinic acid. It showed favorable influenceon pain of alkaline pattern, and exacerbatcdpain of acid pattern, but
had no other effects.Heptylamine has been used by others as a hypertensiveagent.In our studies,its hypertensiveactivity appearedto be weak
and transitory.
The study of lipamineswas the startingpoint for an entire seriesof
into the biologicalrole of the amino group, especiallyas it interresearches
venesin a complexmolecule.We have seenthat, like all other polar group6,
the amino group will act as a fixinggoup in a molecule.Its characteristics
appearto be relatedto its capacityto bind the moleculeto other molecules
in a relativelystrongerand more specificway than other polar groups.
The rcalization of more complex chemical polymers as biological entitiesevidentlyis relatedto this capacityof the amino group, This appean
clear when the amino goup is bound to various acidsin the alpha position
p H A R M A c o D y N A MA
T cc r r v r r y ( p r n r r w o )
391
to form the alpha amino acids that enter into the formation of complex
proteins.In amino sugars,the amino goup showsthe same property, producing the polymer formationscharacteristicof connectivetissues.Furthermore, it is the amino group, acting as a secondpolar group, which givesto
alkaline amino acids their fundamental role in the biological realm, as
mentioned above. The alkaline amino acids,like other amino acids,form
polymers through their amino acid group. However, as these polymers,
histonesor protaminesremain reactivethrough the terminal alkaline nitrogen-containinggroups and it is through these groups that they realize new
bonds, such as to nucleic acids,
In a molecule with two polar groups far apart the amino group will fix
the molecule, while the other active radical wi.ll provide reactivity. The
selectivefixation upon certain constituentsin various placesin the organization accomplishedby the amino group localizesthe intcrventionof the
other active groups of the molecule.
we have consideredmany of the biological substancescontaining an
amino group and a secondactive group. Typical examplesare the local
anestheticsin which the amino group acts to fix the molecule, while the
other energeticformationsintervenemore activelyto induce the anaesthetic
effect. Similarly, in epinephrine and ephedrine,the amino group serves
to fix the molecule,while the hydroxyls later intervenemore specifically.
In somemoleculessuch as the alkaline amino acids,the secondactive
group can be another amine or another nitrogen containing group. It was
for this reasonthat we first becameinterestedin investigatingnatural and
syntheticcompoundswith an amino acid to servefor fixation, and another
energetic center to intervene more actively. We studied amino-butanols,
but no particular activity could be found. These agents, however, were
not lipoids. We were consequentlyinterestedin substanceshaving a lipoid
character as well as two active polar groups of which one is an amine. A
study of theseagentsis now in progress.
Procaine
In this group of agentswe studiedprocaine.We becameespecially
interestedin procaine after we had seensome casesof ulcerationsdue to
varicoseveins healed in a few days with only two intra-arterialinjections
of procaine accordingto Leriche'smethod. With the idea in mind that an
we studiedprocainebase.
actionthroughits lipoidic propertiesintervenes,
procaine
in
base sesameoil, or suspensionin Tween or gum
Solutionsof
we
cellulosewere prepared.Parallelwith theseprocainebasepreparations,
srudiedalso the hydrochlorideas well as severalsaltsof procaine,such as
392
nESEARcH rN pHysIopATHoLocy
lactate,glycerophosphate,
where a further effect was sought through the
acidsat which procainewas bound. We studiedall theseagentsbut only
underthe aspectof their activityin relationshipto the dualisticoffbalances.
No specialeffectswere seenupon virusesand nricrobesfrom this point
view.
of
Injectedsubcutaneously
to rabbits,it was seento increasein the
skin correspondingto the place of injection,the manifestations
of a subsequentsmallpoxinoculation.Added to the medium for culturesof tetrahymenapyriformisin higherdoscs,it led to the appcarance
of almostround
forms. we studiedthe survival time through the capillary tubes method
mentionedabove.For progressive
amountsof procainehydrochlorideadded
to a culture of tetrahymena,no immediateeffect of thc life-spanof the
culturecould be seen.In anothergroup of experiments,the procainewas
added to the culture before its inoculationand thc life-spanof the tetrahymenagrown on this medium was studied.Under thesecircumstances
a
prolongationof the life of the cultureitselfcould bc seen,but only in the
cultureswhich were showingalso a slow growth.
Procainein general,and much more evidentin the basepreparations,
was seento reducethe pH valuesof the secondday wound crust. In micc
and rats, the lethal doseswere seento bc manifestcdthrough the appearanceof convulsions.
Sublethaldoseswcre seento lead to convulsionsafter
a few successive
daysof administration.
In humans,an intramuscularinjection with procainebasewas seento influencepain but this effectwas appare n t l yl i m i t e do n l y t o t h ec a s e sw i t h a n a l k a l i n ep a t t e r np r e s e n tT. h i s a n d t h e
fact that the effectswere no more manifestwith procainebase than with
other lipoidic preparationswith positivepolar group, even when administeredin relativelyhigh dosessuchas of 2 cc. of the 57o solutionof procaine
in oil, indicatesthat its actionother than at the placeof the injection,has
to be relatedlargely to the interventionof procaineas lipoid. A special
mention,however,has to be madefor the anginapain which seemsto be
more evidentlyinfluencedby the injectionwith procainebase than other
pains.
The immediateinfluenceexertedupon systemicanalyses,
if any. was in
generalreducedeven aftcr the administrationof | 0 cc. of. ZVo procainc
h y d r o c h l o r i d eI t. w a sm a n i f e s t epda r t i c u l a r l uy p o nt h e u r i n a r yp H . i n d u c i n g
a changetoward more alkalinevalues.This effectwas transitoryand especially seenfor the first injections.With rcpeateddoses,this effectdecreased
evento the point of disappearing.
T h e s t u d y o f t h e p h a r m a c o l o goyf p r o c a i n eh a s e m p h a s i z e tdh e c h a r acter of its activity as a lipoid with a positivepolar group rathc'rthan a
s .h i s v i e w i s c o n f i r m e db v t h e
d i r e c to n e t h r o u s hi t s c h e m i c acl o n s t i t u c n t T
p H A R M A c o D y N A M T cA c r r v t r y
(e,rnr rwo)
393
fact that the sameeffectsare not obtainedwith any one of thc two constituentsof procaine, para-aminobcnzoic
acid and diethylaminoethanot,
administeredseparatedor evcn togethcr.The only differencein this case
betrveenprocaincand its constituc'nts
residcsin the propertiesof procaine
as such, which we relatedlargely to its lipoidic properties.
Furthermore,the studyof the diflerenteffectsinducedby procainecan
be seen to result from a nonspecificeffect in which these agentsintervene
indirectlyupon the metabolicprocesses
whichoccurat the cellsfor instance,
rather than to a direct influenceexertedby procaineconstituentsupon
specificmetabolicprocesses.In this group would enter the effectsseen
upon different enzymatic processesindirectly influenced by procaine
throughthe nonspecific
changesinduced.
wc have investigatedundcr this specialaspcct the use of procaine
hydrochloridcin solutionswith a pH between3 and 4, as indicatedby A.
Aslan,againstthe manifestations
of old age.The low pH seemsto intervene
by preventing,as long as possiblc,the enzymatichydrolysisof procaineand
thus permittingits absorptionand action as a nondissociated
lipoid. we
found that the protectionexertedby the low pH would result from the
chemotacticnegativeinfluenceexerciscdby hydrosolubleacids upon the
whichthuswill reducethe interventionof the leucocyticenzymes
Ieucocytes,
in the processof hydrolysis.
Furthermore,the fact that the bond of procaine,as such, takes place
throughthe aminoradicalof the P A B A explainswhy a high aciditywould
preventthis local bond of procaincto occur at the placc of the injection
and thus would favor its action at other sitesin the organism.This action
is confirntedby the fact that the analgesicpropertiesof procainedecrease
through the low pH, while at thc same time, its other metaboliceffects
increase.
The fact that good effectswere observedwith procainein old age and
a r t h r i t i s .s c h i z o p h r c n iaan d m r n y o t h c r c o n d i t i o n si,n d i c a t c st h a t a n o n specificaction would take place. The fact that the effectsobtained are
similar to thoseobservedin the sameconditionswith other lipoidic agents
with a positivepolar group.pernritsus to seeas principalfactorin the pharmacologicalaction of procainc, its nonspecificinterventionas positive
lipoid. Besidesthis nonspecificlipoidic effect,some othcrs relatedto its
chemicalconstituentsand rvhich would rcpresentspecificadded factors
Thc relationto thc folic acid would be an example.
haveto be considered.
led us to study variouspreparationsin which
All theseconsiderations
was
procaine
bound to differentagents,with the aim to enhanceits nonspecificintervention.Through procaine lactate,malate and citrate, the
anl
itfi-FlIrjj*:clryr
;.is\r,
394 /
n E s E A R c Hr N p H y s r o p A T H o L o o y
chemotacticnegativeeffect was highly increasedand procaine activity enhanced.Bound to maleic or citraconicacids,the anti-fatty acid activity of
procainewas further directedtoward influencingmore specifically,the abnormalchangesoccurringleadingto the appearance
of abnormalfatty acids.
Tle Elements
with the recognition that elementsact biologically in two opposite directions,we becameinterestedin those with anti-D character,capable of
intervening in offbalance D. we studied the direct action of these agents
upon fatty acids, as well as their indirect action upon processesand substancesrelated to the metabolic changescharacterizing the offbalance D.
we separated these inducing elements-according to their series and
the compartments where they predominantly exercisetheir influence. The
following table showsthis systematization.
T,rsla XIX
InducirrgGroup or Anti-D Group
Metals
Non Metals
Series
Seric.s
Compartments
10_ I V B I V I B , V I I I
Organism
Li l
Metazoic
Cellular
N, I
K l TiiC.
Fe
Ni
Zn
Nuclear
Rb
Zr
Mo
Ru
Pd
cd
Submorphologic Cs
Hf
Os
PtlHe
Primary
VIII'
IIIA
_l_-1-
AI
vA furro
F" I
P l c r
Ca
As l B r
Tl
Bi
Fr
Eu
Am
Tb
Bk
Ho
Tm
Lu
.irffifi$i
p H A R M A c o D y N A MA
T cc r r v r r y ( r r n r
rwo)
395
396
nESEARcHrN pHysropArHoLocy
sodium also favors the appearanceof thiamine-inducedconvulsionsin animals.In rats and mice kept on high salt intake,the convulsantdoseof thiam i n ef e l l f r o m a r o u n d1 5 0 m g r . / l 0 0 g r a m st o b e l o w1 0 0 m g r . / 1 0 0g r a m so f
body weight. Administration of sodium-in the form of sodium chloride
and especiallyas sodium lactate-favorably influencesthe state of shock
which, as we have seen,correspondsto a changetaking place principally
at the metazoic level. In superacuteand acute shock, we consideredthe
excessof sodiumpresentat still lower levels,suchas cellsand tissues.to be
one of the pathogenicfactors.The anomalylies not alone in the excessbut
in the fact that the excessis at a levelto which the cation doesnot belong.
The study of sodium mctabolismin thc light of organizationalsystcmatizationof elementshasrevealedthe importanceof two factors;the combinationwhich is "proper" for an elementin its normal compartmentand
distributionof the elementamongcompartments.
In abnormalconditions.
the unusualcombinationsoccur. At the metazoiccompartment,the bond
of sodiumto chloridecan be consideredto be normal. we have seenthat.
when this combinationdoesnot take place,the result is an anomalycharacterizedby accumulationof excessamountsof sodiumin the immediately
higher compartment.
In stateof shock,for example,the pathogenicanomalyin fatty acids
leadsthem to bind the chlorideion. This removesthe normal combining
factor for sodium which then passeson to the immediatelysuperiorcompartment,the gastro-intestinal
tract, This explainsnot only the excessivc'
passage
of sodiumto thc duodenumbut also its retentionthere. The fact
that sodium is present in reduced amounts in blood and in excessive
a m o u n t si n t h e d u o d e n u mi n t h e s t a t eo f s h o c k .i l l u s t r a t etsh e r u l e d i s c u s s e d
abovewhich we believegovernsthc distributionbetwcenlevelsof elements
in abnormalities.
Potassium
This same rule would explain the distributionof potassium,another
elementof the same series.Potassiumis the principal anti-D cation for
t h e c e l l u l a rc o m p a r t m e n ta, s i t i s o n c o f t h e p r i n c i p a lc o n s t i t u e n tosf t h e
earth'scrust, the environmentin which the nuclei developed.Excessof
potassiumin the cell resultsin a cellular A offbalancewith consequent
active proliferation.Potassiumin excessappearsthus to be the cellular
growth-inducing
factorand its role in cancerhasto be considered
especially
the
at
cellular level. Through the inducedgrorvth,the excessof cellular
potassiumwould thus representthe factor immediatelyresponsiblefor the
invasivephase.
,,|:j:
*dF.i:
p H A R M A c o D y N A M T c^ ( : T t v l r y ( p e n r
rwo)
397
F o l l o w i n gt h e r u l c o f d i s t r i b u t i o no f e l e m e n t sb c t w e c nt h e l e v e l sa n d
c o m p a r t m e n t sa. n e x c e s so f c e l l u l a rp o t a s s i u mw i l l r c s u l t i n l o w b l o o d
potassium.This permits us to associateexccssof cellular potassiumand
hypokalemiain cancerwith activecellularprolifcration.The oppositeoccurs in the stateof shock and in offbalanceD. The amount of potassium
decreases
in the cells. Consequently,
the elementaccumulatesand is retainedin excessivc
amounlsin the compartmcntimmediatclyabovethe cells.
the metazoic.Teleologicallyspcaking,whilc the excessof sodium in the
intestinesand the excessof potassiumin blood could be eliminatedeasily
by means availableto thc organism,they are kept in high amounts in
thesesuperiorcompartmentsas rcscrves,disposablewhcn and if the abnormality disappearsand they can be used properly again.We have seen
horv this samemechanismexplainsthe exccssivcamountsof coppcr in the
blood in cancerpatients.
This redistributionbetweencompartments
explainsanotherfact about
potassium.An excessof potassiumis found in cancercells: the greaterthe
degreeof malignancy,thc g:reaterthe excessof the element.Along with
the cellular excess,the anrountof potassiumin the blood drops to low
values,even below 4 mEq. Thc low blood valuecannot be relatedto lack
of potassium,sincequantitiesof the elementsare eliminatedin the urine.
It has to be consideredas a kind of defensethroughthc higherlevel to an
excessive
amount of the elementat its propcr level.This relationshiphas
led to the comparativestudy of thc potassiumcontent of red cells and
serum as a meansof obtainingindicationsconcerningthe interventionof
this metal at its propercellularlevel.The changesin the potassiumcontent
of red blood cclls are consideredto pararllelthoseoccurring in the cells in
g e n e r a l I. t i s n o t t h e r a t i o b e t w e e nt h c s ev a l u e sw h i c h i s o f i n t c r e s t b
, ut
each value by itself.Low amountsin red cellsand in serumcorrespondto
a quantitativedeficiency;high amountsin both, to an excessof the mctal;
low amountsin cellsand high in serumindicatea metabolicanomalycorrespondingto a depletionof potassiumin the cells as seenin offbalanceD;
while a high amount in cells and with low valuesin thc serum indicatea
cellular offbalanceof the typo A.
Administrationof sodiumand potassiumas therapeuticaids has to be
guided by thesefindings.Isotonicsalineappearsto be adequateas a replacementproduct but it is uselcssto administerit only becausehyponatremia exists, except if this hyponatremiaresults from a quantitative
perspiration.
as in excessive
insufficiency,
The problem to be consideredis
the
normal
bond
for
to
restore
the
cation
at its proper level. Often,
how
w h a t i s n e e d e di s c h l o r i d ei o n s f o r s o d i u m ,n o t m o r e s o d i u m .S i m i l a r l y ,
398
REsEARcH tN pHyslopATHoLocy
p H A R M A c o D y N A M t cA c r r v t r y
(r,rnr rwo)
399
causeof the insolubility resultingfrom the high atomic weight of this element.
In the VI B series, we know little about chromium. Molybdenum
appears to be an active agent. The influencexerted by molybdenum is
neutralizedby the action of methionine,with its active thiol group. Excess
of molybdenumfound in somepasturesinducesa deficiencyin copper and
calcium in anima.lsfollowed by osteomalacyand bone fractures,just as it
induces low fertility. (174) The antagonismbetween these elements is
shown by the inhibitory effect of ammonium molybdateupon oxidaseactivity of ceruloplasmin,the form in which copper is bound to protein in
blood serum. (175)
The anti-D effect of molybdenumis especiallymarked in microbes,in
which it induces morphologicaland tinctorial changes.Bac. anthracis
treatedwith ammoniummolybdateshowsa cocciformchangeand abnormally intensiveGram positivestaining.
Iron
Iron, a member of the VIII seriesand an anti-D agent belongingalso
to the cellular level, is of specialinterest.Its form of activity as cytochrome
oxidaseor hemoglobinhas facilitatedthe understandingof its interveotion.
We could thus relate the high or low amountof the red cells in hemoglobin
which correspondsto the amount of iron ions to thc respcctivooffbalance.
Hypochromia correspondsto a type D offbalance and hyperchromia to a
typ A. These patterns, which were first recognizedthrough clinical investigationsare in accord with the anti-D characterof iron at the cellular
level where it belongs.The existenceof offbalancesof A and D types in
cancer explainsthe high and low values of Fe in the immediatelyhigher
level, the blood serum. Fever, as we have noted, correspondsto an offbalanceof type A at the metazoiccompartment.The fact that serum iron
is low during fever would indicate increasediron activity at the cellular
level, which accordswith its A-inducingrole. We administerediron compounds to cancer patientswith hypochromicanemiato correct the anemia
and also becauseof iron's anti-D effectwhich might act antagonisticallyon
sulfhydryl groups.In severalcases,with very high sulfhydryl index values,
ferrous sulfate has been administeredin dosesas large as severalgrams
daily. In addition to producing an increasein hemoglobinand color index,
ferrous sulfate in large doseshas been observedto have another effect in
some cases,reducingpain of an alkaline pattern.The salutary effect upon
pain was noted more often with reducediron than with ferrous sulfate.No
other influenceupon systemicchangeswas observed.
'irffi
400
nESEARcHrN pHysropATHoLocy
p H A R M A c o D y N A M T^cc r r v t T y ( p , r n r r w o )
401
4OZ /
xESEARcH rN PHYSToPATHoLocY
dosesof 1500 r. In differentexperimentsin which 85 to L)OVoof the controls died in lessthan twelvedays,the mortality rate for animalsgiven zinc
rangedfrom 25 to 50%. In one g'oup, a mortality rate of ISVo was observed.There was a much weakereflect when nickel was incorporatedin
food. No effect at all could be obtainedwith iron.
Mercury
Mercury, a nrember of the IIB series was studied. Theoretically, it
shouldact upon the sulfhydrylgroups,and thus limit the processes
in which
thesegroups take part.
A seriesof compoundsin which mercury is presentin the anion, and
which are routinely utilized as therapeuticagentsbecauseof their diuretic
action and their effectson clectrolytemetabolism,was investigated.Because
of their well-known diuretic effect,we initially used them in patients with
generalizedanasarca.We extendedtheir use to caseswith localizededema
which could be related to tumors with local alkalosis.It was noted that
the mercurial diuretic, in addition to influencingwater and sodium excretion, changedother systemicanalysestoward type A, although only temporarily. However, when these substanceswere administeredover long
periodsof time, other effectswere observedwhich could not be attributed
to diuretic action. In animals with slowly growing tumors, mercury was
found fixed with a degrec of selectivitywithin thesc lesions.After treatment for a certain time, peritumoralfats becamerich in mercury, as recogniz*d macroscopicallythrough abnormal ash color and confirmed by
histochemicalanaiysis.It was interestingto note that mercury appeared
largelyin the necroticpart of tumors.A beneficialinfluenceupon evolution
of tumors was seen,if a type D offbalancewas present.
Bismuth
Bismuth, from the VA serieswas studied.We utilized available antilueticcompounds.Again localizationin peritumoralfats was seen,with the
fats this time becoming abnormally reddish. Neither in animals nor in
patientscould other important effectsbe recognized.
Arsenic
Arsenic, from the same VA series,belongsto the cellular compartment. Its manifest A inducing or anti-D eftect was noted in all compartments if the amount administeredwas high enough. It is interestingthat
Bac. anthracis,under the influenceof arseniousacid, changesto cocci
.'ilru-ff
.,
. i,$i(
i df,,
p H A R M A c o D y N A M TA
ccrrvrry (r,rnr rwo)
4O3
404
IESEARcH
rN
pHystopATHoLocy
p H A R M A C o D y N A MAI cc r r v r r y ( e , r n r r w o )
405
taining whether the eflects of iodine were related to one of the two offbalancespresent,and if this was so, to try to take therapeuticadvantage
of it. Iodine was administeredprincipallyin the form of potassiumiodide
solutionsand Lugol's solution.In most caseswith alkaline pain, the intensitydiminishedand pain evendisappeared
soon after iodine administration. In severalcases,dosesas low as 3-10 drops of Lugol's solutionwere
sufficientto induce such an effect for hours. However, larger dosesor repeated doses produced cffects that wcre distinctly undesirable.Edema
within the neoplasticlesionwas increasedby the administrationof iodine,
sometimesto such an extentas to requirediscontinuingits use before any
other changesin the tumor could be noted.
Researchalso was done with lipid and lipoid moleculesincorporating
iodine. The purposewas to determincwhethcrthesemoleculeswould act
more selectivelyupon abnormal cclls and induce local toxic effects.9,
l 0 - d i - i o d o s t e a r iacc i d a n d 9 , 1 0 , 1 2 . l 3 - t e t r a - i o d o s t e a rai c i d w e r e p r e paredand testedin animalsand humans.No differcnccs
wcre notedbetween
thesesubstances
and inorganiciodinc preparationsin influenceupon syst e m i c a n a l y s e sp, a i n o r t u m o r g r o w t h , i n b o t h a n i m a l s a n d h u m a n s .
Oxygen
Oxygen, an agent with a ncgativeanti-A charactcr,at the organism
level,acts like an anti-fattyacid agentat the lower tissularlcvel. For that
reason,we will discussit here.
At the beginningof our work, we werc intercstedin determiningthe
relationshipbetweencellularmembranepermeabilityand the pathogenesis
of the two offbalances.
If a changein the permeabilityof cell membrane
was the primary mechanisminvolvcd,changingthe oxygentensionin or
around entitieswhere offbalancesoccur misht result in thc correctionof
the abnormal manifestationsprcsent.
Clinical studieswere made in which oxygen was administeredto patients with acid pain pattern.The pain was not relievedas expected.Indeed, its intensitywas even increased.These experimentsindicate that
impairedcell permeability.if it exists,is not thc major factor in the pathogenic mechanisminvolvedin the acid pain pattern.Actually, thesestudies
indicatedthat another pathogenicmechanismwas involved since oxygen
administrationincreasedthis pain. According:ly,
it had an oppositeeffect
the pain being
on alkalinepain. The intensityof alkalinepain decreased,
often entirely relievedby administrationof oxygen.We found that pain
producedby traumatic injuries.which was subsequently
identifiedas inpattern,
the
alkaline
could
be
satisfactorily
variablyof
relievedby oxygen,
406
xESEARcHrN pHysropATHoLocy
Such relief occurred in patients who had suffered all sorts of traumatic
injuries, from superficialwounds to severecomminutedfractures.
A curious phenomenonwas seento occur which limited the practical
usefulnessof oxygen. After fust relieving pain, continuation of oxygen
administrationled to appearanceof a new pain. The patient was able to
distinguishbetweenoriginal and new pain by is localization,by the different quality of the sensation,and also by the fact that insteadof being releved by oxygen, the new pain tended to increasewith the continued
administrationof oxygcn.It disappeared
soon after oxygenadministration
was discontinued.If oxygen were administeredagain, the new pain returned within a short time. The new pain might becomesevereand even
unbearablervith continuationof oxygenadministration.
on the other hand,
when administrationof oxygen was stopped,the original pain again appearedwithin l0-20 minutes.
The appearanceof a "new pain" and the resemblancebetween thcse
changesand those observedwith the use of lipids, suggesteda change in
the pain pattern itself, although the changesinduced by oxygen evolved
over minutesinsteadof days.This was confirmedby following the responsc
to acidifyingand alkalizingagents,the only adequatemeansto investigate
the patternin thesecases.By this test the new pain was found to be of an
acid pattern.
Bccauseof the possibilityof inducinga period of calm betweenthe
old alkalinc and new acid pain, oxygenadministrationstill seemedto bc
useful.In casesof traumaticpain, alwayswith an alkalinepattern,successful resuls were obtained.A necessary
condition appearedto be the physical
and mentalability of the patientto guidethe administrationof oxygen.He
had to recognizeif too little or too much was being administeredon the
basisof the di{Iercntsensations
felt, and consequently,
to adjust the administrationto the optimum amount. Extreme pain causedby extensive
traumatic injuries was controlledvery successfullywhen the patient could
be taught to utilize oxygenproperly.
In addition to pain relief, an effectupon cvolutionof the lesion iself
was manifcst.In a few dayswith this form of oxygentreatment,guidedby
the subjectivesensations,
thc healingprocessitself was observedto be sufto
make
ficiently advanced
the pain disappearentirely. Healing of the
woundsseemedto be greatlyenhancedby oxygen therapyguided by the
patient.In severalcasesof open comminutedfracturesin which amputation was considcrcdinevitableon admission,unexpectedimprovementwas
noted. Atonic rvoundswere transformed,becomingrich in granulations.
and hcalingwas rapid.
p H A R M A c o D y N A MA
T cc r r v r r y ( p , r n r r w o )
40j
408
R E s E ^ R C Hr N p n y s t o p A T H o L o c y
pHARMAcoDyNAMTc Acrtvlry
(p,rnr rwo)
409
410
xESEARcH rN pHysropATHoLocy
p H A R M A C o D y N A M t cA c r r v r r y
(nrrnr rwo)
4lI
TrsLr XX
CLrlrc,rLRssulrs Wtrtr AcrNrs Tur
T H e O n n s , { t - A N c ET y p t
Group
Agent
.,A"
Systemic
Level
Pain
None
Slight
None
Fair
None
Slight
Fair
Slight
Fair
Oleic
SIight
None
Slight
Slight
Good
Good
Acr Upox
lnfluenccsExerted
Upon Tunror
None
Some,not consistent,
not persistent
Good I
Fair
Fair, not consistent,
not persrstent
.:
rr
{t
ta
None I None
I
None
Some, not consistent
not perslstent
None
I S l i g h t ,n o t c o n s l s t e n t ,
not persistent
Fair
Good l S l i g h t F a i r
Good I Cood I F a r r
None
None N o n e
Methyl thioglycolate
Tetrahydronaphthalene oersulfides
Srk"lr^
| Alkyldiselenide
Cctmpoundsl
I Perselenide
-l
Peracids
Perborate
Perchlorate
Slight
Slight
Fair
Fair
Slight
Fair
Fair
Fair
Fair
Slight
Fair
Cood
i=;-
1-'F
---.l
Hormones
i Testosterone
Slight
None
Mustards
I Sulfur mustard
Fh"
Fair
Fair
to good
Slight
Hydrines
Epichlorohydrin
Ciood, consistent,
persistent
Fair, not consistent,
not perslstent
Good, consistcnt
and pcrsistcnt
Good, consistent,
.f".ti:,"n,..
S o m e ,n o t c o n s i s t e n t ,
not persistent
4t2 /
RESEARCH IN
PHYSIOPATHOLOGY
Trsl-e XXI
Cr-rNrcrl Results Wtrs AceNts THr.r Acr Upox
THe OnrsrLANcETYpe "D"
Group
Sterols
Agent
Cholesterol
Insapon.fraction
of organs
of eggs
of milk
Alcohols
Octanol
Slight
Polyconjugatcd
Crotonic
Ricinoleic
Salicylic
Hormones
Estrogens
Amines
Aminobutanol
Hexylamine
Heptylamine
Glucosamine
Halogen
Iodine
i Oxvcen
'|
Fair
aa
ar
ra
Good
Fair
Slight
None
None
Slight
Good
None
None
Slight
Slight
Fair
F^t;:
,,
Slight
Fair
Slight
.:h
a,
Good
Good
Fair
iI N"*
:.
.:
None
None
Slight, not consistent,
not persistent
,: .:
Slight,
r'
Fair,
aa
aa
ra
aa
Slight
Fair,
None
Good
None
Slight
None
None
Fair
Good
Good, consistent,persistent
None
ra
Slight
aa
Good, consistent,persistent
Fair, not consistent,
not persistent
Slight
None
t?I'
Niketamidc
Iron
Mercury
Bismuth
lnfluences Exerted
Upon Tumor
aa
Good
Fair
Inositol
U nsaturated
Oleic
Linoleic
Polyunsaturated
Metals
Fair
Aliphatic saturated
Butanol
Pentanol
Heptanol
Octanediol
Nonanol
Polyalcohol
Glycerol
Nicotinic
acid deriv.
Systemic
Level
None
"
aa
C H A P T ELR1
1 ' H E R A P E U T I CA P P R O A ( ] H
ln
lLHe
sruDrESpREvroust-yDEscRrBEo
have identifiedmany factors involved in the pathogenesisof abnormal conditions and have shown that
there are similar factors which also govern the pharmacologicalactivity of
variousagents.Out of this peculiarrelationshipbetweenabnormalconditions and agentshas evolveda therapeuticmethodwhich is fundamentally
different from methodsin common use.
In conventionaltherapeuticefforts againstdisease-in chemotherapy
of cancer for example-methods as standardizedas possibleare sought.
The goal is to find agentscapableof influencingthe cancercells,if not in
all tumors,then at least in tumors of the sameorigin. Within the limitations imposedby the generaltoxicityof thc agent,the conductof treatment
is standardizedas far as possible.The criterion of value of a treatment is
its ability to overcomeindividual variations.
In our approach,in clear contradistinction,the effort is to influencethe
complexcondition as it is presentat differentlevelsof the organization.It
correspondsto treatment individualizedas far as possible.Therapeutic
effortsare guided in all their aspects--{hoiceof agents,dosesto be usedwith their multiple quantitativeand qualitative
by existingmanifestations
variations.The fact that many of thesevariationsoccur during treatment,
as a responseof the organisminducedby the medicationitself, increases
the individual characterof treatment.No predictioncan be made about
the whole course of treatmentto be used for an individual patient becausethe individualclualitativeand quantitativechangeswhich occur during the treatmentcannotbe foreseen.
414 /
n E S E ^ R C Hl N
pHysropArHoLocy
THERAPEUTIC
APPROACH
4I5
416 /
xEsEARcH tN pHyslopArHoLocy
i;riiNF]
T H E R A P E U T T CA P P R O A C H /
417
418
xEsEARcH rN pHysropATHoLocy
$:in
,ii ;
:J...i,
THERAPEUTIC
^PPROACH
,/
419
420
nESEARcHtN pHyslopATHoLocy
THERAPEUTTc APPRoACH
421
422
xESEARcH rN pHysropATHoLocy
THER^PEUTIC
APPROACH
423
Trauma
A particularly interestingapplicationof the method describedabove is in
the treatment of pain related to traumatic lesions,The study of wounds
from the standpointof the offbalancepresenthas revcaleda definite pattern in their evolution. In all wounds, there is an initial period during
which the offbalanceis alwaystypc D, correspondingto a predominanceof
fatty acids. This can be recognizedindirectly by measuringthe local pH
which shows alkalosis,the chloride content which shows a manifest increase,and by cytologicaland histologicalstudieswhich indicatea rapid
cellular agtng process, with necrosis and sloughing. In uncomplicated
wounds,in humans, the painful period usually lasts a few days. The pain
in this initial period is of an alkaline pattern.
The constancyof the alkalinepatternin pain in traumaticlesionselimi-
:let
424
RESEARcH tN
pHysropATHoLo6y
W'TH
BUTYL
ALCAHOL
WITH
PHYSrcLOGICAL
SAL'TVE
4O cosas
25 cosas
t 2 . 5 4 Ar.9 5 0 . )
T H E R A P E U T I C^ P P R O A C H
425
WITH
BUTYL ALCOHOL
362 coses
fvo
TREATMENT
585 coses
9t%
+
O = minimol or no po,n
426
xESEARcH rN pHysropATHoLocy
Burns
The pain of burns has been successfullytreated with higher alcohols such
as butanol, heptanolor polyunsaturatedalcohols.However, here the prob.
lem is not so simple since pain is only one, and not the most important,
of the manifestations.Systemicfunctions are often so greatly impaired as
to representthe immediatecauseof death.The higher alcoholsare effective
in controlling pain and local troubles,while other agentsare neededwhen
the condition also involves the systemic level. Glycerol and especially
sterolswhich are presentin the insaponifiablefractions of organs such as
.,i,i:.
4:li,r,i
T H E R A P E U T T CA P P R O A C H /
427
428
nesEARcH rN PHYsroPATHoLocy
Vertigo
Another interestingapplicationof the same therapeuticmethod has been
in otological conditions.B, Welt has made a stimulatingcontribution
to the study of ear conditionsin terms of the pathogenicmechanisminvolved and an even more valuablecontributionto therapy.
Vertigowas one of the conditionsstudied.In a few cases,by comparing
concomitantvariationsin the intensityof the symptomand changesin the
urinary pH, we had been able to show that acid and alkaline pathogenic
patternsexist,similar to thoseseenin pain.
Going on from there, B. Welt, in an extensivestudy, has shown thc
presenceof this dualism and has successfullyapplied the therapeutic
methoddiscussedabove,to a significantnumber of casesof vertigo.The
major advantageof this method over other treatmentsfor this condition
lies in its extremesimplicity.
For vertigo,which has so many difTerentcauses,an etiologicalapproach
althoughideal,has seemedimpossibleto all workersin the field especially
as a routine procedurein medical practice.An initial simplificationof the
approachcould be made by relating vertigo to acid and alkaline patterns.
thus reducingtreatmentchoiceto two groupsof agents.Welt, in a further
step, simplified even this procedure,making the method highly applicable
in routine medicalpractice.He administercdto patientsone or the other
of two agentschosenfrom each group, such as butanol or sodium thiosulfate,being guided in his choice by clinical aspectssuch as changesin
the symptomwith the time of day or the intakeof food. A favorablechange
in the symptomwas consideredto be a confirmationof this tentativediagnosisof the pattern.The treatmentwas then continuedwith the samegroup
of agents.An increasein vertigo led to the use of the opposite group of
agents.The clinical resultsobtainedby Welt with this simple method arc
impressive.(190) He arrivesat the following conclusions:
" Summaryand Conclusions
A seriesof 44 patientshaving the symptom of dizzinesshas been
studied.
The symptom of dizzinesshas been analyzed according to Revici's
conceptconcerningthe alkaline or acid patternof the symptoms.
In 80% of the analyzedcases,the responseshave shown the existence
of an alkalineor an acid pattern;lTVo showedan inconsistent
relationshipl
8% showedno result.
T H E R A P E U T T CA P P R o A c H
429
(lel)
" Sum mary and C oncl usi ons
A seriesof 106 caseswith the symptomof vertigohas beenpresentedin
this and a previouspaperand analyzedaccordingto Revici'sconceptof a
metabolic imbalancewhich exists in pathologicalfoci and manifestsitself
as a local alkalosisor acidosis.
Sevencaseswere elinrinatedfor lack of data. This left a total of 99
case studies.This figure represents
the sum of 42 casestermed the 1953
seriesand the presentseriesof 57 cases.The two serieswere combined
and analyzedaccordingto sex, alkalineor acid characterof the symptom,
and clinicaldiagnosis.
The analysisshowed an averagegood result of. 80% for both series.
This percentageindicates a consistentcorrelation of the use of Revici's
method and substanceswith the results achieved.Additional substances
from various sourceswere utilized in the present study. It is here again
emphasizedthat I have submittedthis article for the practical reasonthat
the method describedis simple and has bcen successfulin its application.
In addition, it gives a new systematizationto clinical data in the field of
otolaryngology.Finally, this communicationpoints to a vast group of sub-
430
RESEARcHtN pHyslopATHoLocy
T H E R A P E U T T CA P P R o A C H
431
432
RESE^RCH IN
PHYSIOPATHOLOGY
LeftEar-1.1.
.tl
trd
l?l
zJe
In i t i a l
After treatment
tl
?0
I
t
t!
t0
tl
3r
tl
r
t0
||c
Frc. 146,M a n i f e s t i m p r o v e m e n t i n t h e i m p a i r e d h e a r i n g is obtained in a subject
wirh acid pattern, through guided treatment with negative lipoids. (Courtesy of Dr.
B. Welt.)
RightEar- L.L.
.tl
frd
rl
n
I
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n lr r:Innttrnn
tl
--- Inltlal
-After treatrent
tl
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F t o . l 4 6 b i s . I n t b e s a m ep a t i e n ta s i n F i g . 1 4 6 t h e i m p r o v e m e n ti s n o t a s m a n i f e s t f o r
the right ear as for the left ear.
THER^PEUTIC ^PPROACH
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impaired hcaring.
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A U D I O G R A ILTe{ -f t E a r - F . M .
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434
RBSEARcH IN
PHYSIOPATHOLOGY
500tr r00et5001000
t00 |e0t3t0 f00c
lrrrl I
t0
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ItI
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t0
t0
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ts
F l c . 1 4 9 . A n e x c e p t i o n a l l yg o o d r e s u l t i s o b t a i n e df o r t h e l e f t e a r i n t h e c a s e A . W . .
i n s p i t eo f a n i n i t i a l l yv e r y i m p a i r e dh e a r i n g .
5m t50 1000
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THERAPT,UTTA
c PPRoACH
435
Itching
Acid and alkalinepatternsof itching havc beenrecognized,and treatment
basedupon dual pathogenesis
has been applied.The agentto be used is
determinedby the pattern found through analyses.Dosage,as for pain, is
established
accordingto the subjectiveresponse.
Following thesemethods,
many casesof intractableitching have becn completelycontrolled. of thc
group of agentswith positivecharactcr,butanol and heptanolhave produced the best results;in the oppositcgroup, cpichlorohydrinand sodium
thiosulfatehave been most effectivc.
A*hritis
The analysisof the pattern of pain has shown that often in rheumatoid
arthritisthe patternis alkaline,while in osteoarthritis,
it is acid. Comparison of variationsin pain intensitywith changesin urinary surfacetension
helps to determine the pattern. However, with urinary surface tension
values usedas a guide both for agentand dosage,favorableclinical results
have been obtained in casesresistantto other therapy. Arthritis, painful
for many months, was controlled in many instanceswith severaldosesof
heptanol or of sodium thiosulfatewith epichlorohydrinand more recntly
with propionic aldehydeor sulfurizedtetralin. The simplicity of treatment,
total lack of undesirableside effects,and the long period of improvement
436
nESEARcHrN pHysropATHoLocy
THER^PEUTIC APPRO^CH
437
Case $3
M r . N . K . , 5 5 y e a r so l d , s u f f e r e df o r m a n y y e a r sw i t h g e n e r a l i z eadn d
very painful rheumatoidarthritis, Prior to coming under our care, hc
had beenbedriddenfor six months,entirelyimmobilizedwith severepain
in arms,back and legs.Thc patientwas entirelyincapacitated,
unableeven
to feed himself.Treatmentswith differentcortisoneprcparations,ACTH,
g o l d ,e t c , ,p r o v i d e dp r a c t i c a l l n
y o r e l i e fo f t h e s e v e r cp a i n s .H e c a m eu n d e r
o u r c a r e ,e n t i r e l yi m m o b i l i z e da n d i n s c v c r ep a i n . A c c o r d i n gt o t h e a n a l ysis which showcda low urinaryspccificgravity.and pH, high surfacetcnsion and a low blood scrum pot:rssium,
the paticntreccivedmedicationsof
t h e g r o u pA , s o d i u mt h i o s u l f a t ee,p i c h l o r o h y d r iann d h e p t y l d i s e l e n i dUen. der this medicationthc patientmadc a most dranraticrecovery.within a
f e w d a y s ,t h e p a t i e n tw a s o u t o f b c d w i t h o u tp a i n . w a l k i n gn o r m a l l ya n d
w i t h f u l l f u n c t i o n acl a p a c i t yo f h i s a r m sa n d l e g s .W h i l c s t i l l u n d e rt r e a t m e n t
t h e p a t i e n tw e n t b a c k t o e n t i r e l yn o r m a la c t i v i t y .H e g a i n e d3 5 l b s . i n t h e
Iastmonths.
ConnectiveTissueConditions
We have discussedpreviouslythe hypothesisthat corticoidsactively intervenein the synthesisof an cntire group of substances
in the organism.
The energcticformationbetwcenCrr and C3, o[ thesecorticoidswould act
as a mold or templateto attractand keep variousradicalsin specialreciprocal position,thus favoring,as a secondstep,the synthesisof new substances.Thesesubstances
would vary accordingto the cnergeticformation
of the corticoid.In one group,whichcould be callcdthe "gluco" group,wc
identifiedglucose,glucuronicacid, glucosaminc,gaJactosamine,
glucosaminic acid,galactosaminic
acid,and someotheruronic acids.Even ascorbic
acid could be relatedto such a synthesis
mechanism.
we investigatedsome of thesesubstances,
particularlyin relation to
t h e t h e r a p e u t ipcr o p er t i e so f c o r t i c o i d sI.n 1 9 5 1 ,w e c h o s eg l u c o s a m i naes
a substance
closelycorresponding
to the templateof cortisone.Glucosamine
treatmentwas usedin patientswith pemphigus.rheumatoidarthritis.psoriasis,ocularconditionand allergy.The followingcasesillustratethe results
obtained.
Mr. H. S., 46 years old, had generalizedpsoriasis.In the last
twelveyea$, the lesionshad progressed
continuouslyto cover more
thanT5Vo of his skin. Local treatmenthad had little effect.Cortisone
injections cleared the skin in two weeks but not completely. With
438
n E S E A R C Hr N p H y s t o p A T H o L o c y
THERAPEUTTC APPROACH
439
Hemorrhage
In studying hemorrhage,it seemednecessaryfirst to define preciselythe
conditionsunder which this importantepisodeoccurs,as we did for pain,
itching,etc., sincethe coursco[ hcmorrhage,and particularlyits response
to treatment,seemsto dependgreatly upon certain peculiaritiesrelated to
its pathogenesis.Hemorrhageresultsfrom a break in the continuity of a
blood vessel,which can be inducedby an externalinfluenceon a previously
normal vessel,or can occur as a result of proccssestaking place in abnormal vessels.we havc called the first type of hemorrhage"accidental" or
"traumatic" and reservedthe ternr "pathological"for the second,which
apparsto be a direct result of pathologicalchangesin a vessel.The latter
term is used in the samc senscas it is usedfor fractureswhere "pathological" indicatesonly preexistinglesionsin the bones.
In a traumatichemorrhage,the therapeuticproblem is limited to stopping the flow of blood. Mechanicalmeansto close the bleedingvascular
wound or agentsable to increasethe capacityof the blood to form clots
can be used.They representthe only approachin accidentalhemorrhage.
In pathologicalhemorrhage,other problems arise. The knowledgeof
the local pathologicalchanges,which lead to the appearanceof bleeding,
is importantboth for prcventingand controllinghemorrhage.The pathogenic factor involved will be consideredalong with the problem of hemostasis.
H emorrhagiparousA gents
Local and generalfactorsare involvedin pathologicalhemorrhage.Ulceratedlesionsprovide a favorablecondition.So doesloca.linfection.Anticoagulantsmay act indirectly upon blood vessels.
propcrtiesof somesubstances
Hemorrhagiparous
bccameevidentduring researchon their therapeuticuse in cancerpatients.For example:
P. T., a patient with carcinomaof ttre floor of the mouth widely
ulceratedin the submaxillarregion,bled sporadicallyfrom the lesion.
440
nESEARCH tN
PHYSToPATHoLocY
'f r' t,
:.:.5: i
T H E R A P E T J T t cA P P R o A c H
441
Antihemorrhagic A gents
Butanol
I n 1 9 4 3 , d u r i n g r c s e a r c hs t u d i e sc o n c c r n i n gt h e p h a r m a c o l o g i c a l
activity of glycerol on abnormal foci, its hemorrhagiparouseftect
appearedas a serious handicap.Various hemostypticsubstanceswere
testedwithout sufficienteffcct. At this time, a new product, to which a
hemostaticeffect was attributcd.appearcdon the market. It was a very
w e a k s o l u t i o n( a r o u n d l / 1 0 , 0 0 0 ) o f o c t a n o li n s a l i n e .w e c o u l d f i n d n o
therapeuticeflect for it. Howevcr, we were studyingbutanol and other
higher aliphaticalcoholswith lipoidic propcrties,and we decidedto test
butanol for its antihemorrhagicactivity, hoping that it might counteract
the undesirablehemorrhagiparous
effectof glycerol.It did and we have
sinceaddedbutanolto glycerolfor this purposc.
We observedthc remarkablehemostaticeffectof butanolyearslater in
a patient with severehemorrhage,to whom dosesof l0 cc. of a 6.5Vo
solutionwere given intravenously.
Henrorrhagestoppedin a few minutes.
Sincethen, wc have succcssfully
appliedbutanolclinicallyin hemorrhages
of variousorigins.
A s a n a n t i h e m o r r h a ga
i cg e n t .b u t a n o li s a d m i n i s t e r eedi t h e rp a r e n t e r ally as a 6.5vo solutionin saline,or orally as 6.59,i solutionin water.The
r o u t eo f a d m i n i s t r a t i o n - i n t r a v c n o uisn,t r a m u s c u l asr .u b c u t a n e o uosr o r a l
'liijr.t,i!'
442
RESEARcHrN pHysrop^THoLocy
-is
THERAPEUTIC ^PPRoACH
Number of casesobserved
Number of caseswith profuse hemorrhage
Percent of caseswith profuse hemorrhage
Number of deaths attributed to hemorrhage
Percentof deathsin caseswith profuse hemorrhaget
443
Group I
Group II
Untreated
n
-Butanol
- -256-344
l8
25
j
7
lz
l
67
4
444
xESEARcH IN pHysropATHoLocy
few patientswho neglectedto takc the mcdication,and in whom a hemorrhageappeared,bleedingwas rapidly conrrolledby butanol. (Note 6)
ExperimerrtulResearchon the Henro.stl'ptic'
Eflet't; ln a seriesof studies
we soughtto find the mechanismby which butanol controlshemorrhage.
In an investigationcarried out in our laboratory,M. Bier and p. Teitelbaum (194) showedthat individualmembersof the homologousseriesof
aliphaticalcoholsdecreasethc degreeof rctractionof clots when added in
v i t r o t o b l o o d .( F i g s .l 4 l t o 1 4 3 ) B y v a r y i n gt h e a n r o u n t sa d d e d ,r h i se f f e c r
was observedto occur only at valuesabovea critical concentrationof the
alcoholin blood. Bier could show that the critical value.which diffcrs for
nrembersof the homologousseries,is proportionalto the lethal toxic dose
in mice.
This non-retractionof clots in vitro also can be rccognizedin cascs
treatedwith butanol through the gelatinousaspectof the clots at the moment whenthc hemorrhage
s t o p s .H o w e v e r ,i t i s i n t e r c s t i n gt o n o t e t h a t
t h e a m o u n to f b u t a n o li n j c c t e d c, o n s i d e r e idn t e r m so f t h c a m o u n to f t h e
circulatingblood. resultsin concentrations
considcrablybelow the critical
valuesneedcdto producethis effcct in vitro. However,concentrationof
butanolat the sitc of the wound may explainthis. lt is also of interestthat
the samc gelatinouscharactcrtypifiesthc clots which remain attachedto
b l e e d i n gl e s i o n si n a n i n r a l st r e a t e dw i t h b u t a n o l .I n m i c c , w h e n a p o r t i o n
of tail was cut and butanol was used,the abnormallylong clot remained
adherentto the wound,differingfrom that secnin controls.
ln spiteof changesin the clot, it scenrsthat the effcct upon the blood
itsclf, and its coagulationin a wound, representsonly one of the means
throughwhich butanolcontrolshemorrhagc.
The spcedwith which butanol
acts,often within secondsaftcr intravcnousinjcction,is nruch greaterthan
blood coagulationtime. Consequently,
changesin clot formation alone do
not appearsufficientto cxplain the mcchanismthrough which the rapid
henrostasis
occurs.
M. Bier and H. Lerner in our laboratory studied the influence
exertedby butanol upon hcmorrhageinducedby the highly activeprotcolytic enzyme,ficine.(195) Thcy were able to inducestandardized
hemorrhagesby injectingficinesolutionsunder the skin of the abdomenof white
n t i c c .( F i g . l 5 l a ) ( l n o t h e ra n i m a l sa n d w i t h o t h c rs i t e so f i n j c c t i o ni n m i c e .
the individualvariationswere too greatto make the resultingbleedingusef ul as nraterialfor tcsting the cffect of agcntsupon hemorrhage.
) with
adequatcdosesof ficinc, sevcrehcmorrhages
followcd by skin ulcerations
were induccd.The bleeding.and eventhe ulcerations,
were almostentirely
preventcdwhen butanolwas administcred.
Figurc I 5 I b showsthe resultsof
"r'!:l{-i1r!::,
. i . . .
, .
T H E R . { I } I ]U T I C
A PTROAC}I
445
446
xESEARcH rN pHysropATHoLocy
rnlfin,
r:r1
iiii
THERAPEUTTC APPROAcH
447
448 /
R E S E A R c Hr N p H y s r o p A T H o L o c y
T H E R A P E U T T CA P P R O A C H
449
450
REsEARcH rN pHysropATHoLocy
instence,l0 cc. of the butanol solution, 2-5 cc, of the patient's blood is
withdrawn. After mixing them, and without removing the needlefrom the
vein, the contentsof the syringeare injectedintravenously.A similar mixture can be injected also intramuscularlyor subcutaneously.The results
have been very good. In some casesin which butanol alone was not able
to stop a hemorrhage,the blood-butanolmixture did. with this special
technique,we have becn able, in recent times, to bring most hemorrhages
under control within a few minutes.
The fact that agentswith positivepolar groups,such as sterols,glycerol,
coramine,thiamine and others, have hemorrhagiparousactivity led us to
try to influencehemorrhagewith agentsconsideredbiologicallyantagonistic
through their negativepolar group.
Many years ago, experimentingwith cilorine solutionsin saline, we
observeda manifcst effcct upon coagulationtime. Intravenousinjection of
such solutionsbrought coagulationtime to values as low as one minute.
The addition of thesesolutionsto butanol greatly increaseits eftectin vitro
upon coagulationtime but had lesseftectupon hemostasisin vivo. Similar
but somewhatless manifest effectswere obtained by adding hydrochloric
acid to butanol solution. On the other hand, organic acids such as oxalic,
malonic,citric, lactic maleic or citraconicshoweda favorable effect.Without changing coagulability of the blood, these acids were seen to increasethe hemostaticeffect.
Fatty Acids
The same antagonismto sterolsand glycerol led us to use fatty acids
from cod liver oil. (197) While resultsin severelarge arterial hemorrhages
were not impressive,the effect upon oozing capillary, venous and small
arterialhemorrhagcs
was vcry good in a largeproportionof cases.For example:
N. V., 57 years old, with multiple pulmonarymetastaticlesions
from a hypernephroma,
had frequenthemoptysis.
At times,the bleeding becamemore accentuated,the patient expectoratingclots as well
as uncoagulatedfresh blood. Intramuscular administration of two
dosesof 10 cc. of a 6.57o solutionof butanol at half hour intervals
had little influence upon the bleeding.Intramuscular administration
of I cc. of a solutionof l0% of the mixture of fatty acids obtained
from cod liver oil stoppedthe bleedingin less than 20 minutes, with
the effect persistingfor more than two months. A new episode of
oozing bleedingwas again immediatelycontrolledby injection of the
fatty acid preparation.
; i,ii:;
Sillj r;;rlri:ir8
..i.i
THERAPEUTTC APPROACH
45L
Allergic Conditions
Allergic conditions, which as seen above, are related to the defense
mechanism,also have been integratedinto the general therapeutic approach. This has largely permittedus to apply to such conditionsthe same
therapeuticmeasuresused in general,The study of the manifestationshas,
however,shown that exceptfor the anaphylacticshock, the clinical allergy
correspondsto the prolongedphasewith the predominanceeither of sterols
or lipoacids.The urinary surfacetension has appearedas a valuablecriterion to indicate the occurring offbalance.[n caseswith a high urinary
surfacetension,sodiumthiosulfate,aldehydesor epichlorohydrinhave given
particularly good results,while the caseswith low surfacetension showed
favorable responseto heptanol and butanol. Food allcrgies,asthma,urticaria which had persistedfor years and were insufficientlyinfluencedeven
by corticoids-have respondedwith the complete disappearanceof their
manifestationswith an adequatedosc of one or the other of the medications mentionedabove.In most of the caseskept under continuoustreatment with minimal doses,favorableresultspcrsistedeven after the subjects
were again under the influence of antigens,The following observations
illustratethese results.
Mrs. M. S., 45 years old, with skin and eye allergic manifestations,
higbly sensitivefor the past six years to fish, eggs, alcoholic beverages,
some vegetablesand fruits, and especiallyto dogs, showed no favorable
response to corticoids. With high urinary surface tension and high
the patient was treatedwith 50 mg. sodium thiosulfateand
eosinophiles,
2 mg.epichlorohydrin,four times a day. The symptomsdecreasedprogressivelyto disappearin a week. After a month's treatment,she was able
to take-without any inconvenience-foodsand alcohol to which she had
been previouslysensitive.After another two months, the acquisitionof
three puppiesproducedno disagreeable
effects.The patient has continued
:"f-
452 /
REsEARcH rN pHystopATHoLoGy
on the sametrcatment,but reducedto twicc a day, for the past year without recurrenceof the manifestations.
Mr. A. L., 58 yearsold, had frequentattacksof asthmafor over four
and a half ycars which left him unableto work for the past year. Because
of low urinary surfacetension,the patient was put on butanol-S cc. of a
6.5Vo solutionIn wn1s1-to be taken every six hours. Although the first
dosesshowedmarked objectiveand subjectivechanges,the treatmentwas
continued.The patient was free of attacksfor four months at which time
he stoppedmedication.Two weeks after stopping medication,he had an
attack,the first in four and a half months and this was followed by another
the following day. By resumingthe medication,he has been free of symptoms for more than a year.
The possibilityof preventingallergicmanifestations
with butanol,which
appearedso clear in the traumaticlesions,opensan entirelynew view for
many conditions where such a pathogenesis
intervenes.
The conceptof a nervoustissueallergyintcrveningin the pathogenesis
of multiple sclerosis,Ied us to try similar antiallergictreatments.The low
surfacetensionled us to utilizeas agcnts,lipoidswith a positivecharacter.
Interesting-although not constant-results were obtained with insaponifiablefractionof cow brain. Good resultswereobtainedwith butanol-100
mgr. four times a day-together with an antihistamine
preparation.
ARTERIOSCLEROSIS
One of the most important medicalproblcmsfrom all points of viewpathogenic,therapeuticand even social-since it still representsthe chief
direct causeof death in most civilizcdsocieties,
is that of arteriosclerosis.
The fact that lipids seemto intervenein its pathogenesis,
has led us to consider this condition from the point of view of biologicaloffbalances.With
the developmentof our rcsearch,wc tried to apply to this conditiona systematicanalysisin accordancewith the basic conceptspresentedabove.
This attempthas permittedus to arrive at some new views which will be
discussed
briefly here.
The analysisof the specificmanifestationsof arteriosclerosis
from the
point of view of organiTatiorr,
that is, as relatedto the differentlevels,led
us to recognizethat it represents
a conditionprincipallylimited to the level
more
organism,and
especiallyto its secondarypart, the circulatorysystem.
Many of the fundamentalcharactersof the condition could be explainedby
the relationshipof this lcvel to other levclsof the organization,as we will
seebelow.
T H E R A P E U T T CA P P R O A c H
453
From the point of view of the dualisticconcept,it was easy to see that
arteriosclerosis
corresponds
to an offbalancetype A. All the analysesconcerning the systemiclevel show patternswhich indicatc this offbalance.
High urinary surfacetension,low urinaryspecificgravity,high urinary pH,
low urinary sulfhydryl index, found in the routine analysespoint to such
an offbalance.The long persistenceof the skin wheal seen in the subjects
analyzed,confirms this diagnosis.The fact that some manifestations
of
arteriosclerosis
can be induced experimentallyin animals through the administrationof cholesterol,has placedinto limelight the pathogenicrole of
cholesterol.While its administrationin high amountsto rabbitsor cockerels
inducesatheromas,it doesnot inducethe complexconditionitself.Usually
the animalsreturn to normal, even with a rapid healingof their atheromas
after the suppression
of the administration
of cholesterol,Although hypercholesteremia
representsthus only one pathogenicfactor, it appearedinterestingto investigateits interventionin the condition.
The relationshipof cholesteremia
to arteriosclerosis
is certainlynot a
simpleone.The total amountof cholesterol
in blood alone,in its free form,
and the macromolecules
of certain dimensionsrcsultingfrom their bond
to the other constituents
of thc blood, althoughrelatedto arteriosclerosis,
do not seemto representby themselves
the pathogenicfactor of this condition. This latter seemsto be relatedto a more specificinterventionof this
substanceat the level of the blood and circulatorysystcm.Researchin
this field has led us to recognizethis specialintervention.
It is a known fact that the appearance
of crystalsof cholesterolin the
cellsof the intima of the arteriesand in the cellsof the vasavasorumrepresentsan essentialfactor in the pathogenesis
of atheromas.Study of the
capacity of blood serum to dissolveor, on the contrary, to precipitate
cholesterolhas permittedto link it with the appearanceof specificarterial
lesionsin this condition.
In this study, we used the procedureoriginally devisedby Policard
the relationshipof cholesteroland arthritis.Blood is
when he investigated
veinous
punctureand the serumis separatedaseptically.
through
obtained
The amountof total cholesterolis determinedin the blood serum.A sterile
crystal of cholesterolis addedto the serum.The treatedserum is incubated
throughfiltrationfrom
for 6-12 hoursat 37"C. The serumis thenseparated
the crystal added,or from those formed during the incubation.The difference between the amount of cholesterolin the serum before and after
incubationwith the crystal.showsthat while someseraincreasetheir content in cholesterolthroughthis treatment,othersdecreaseit.
We could show that the serum of those rabbits which were fed with
454 /
n E s E A R c Hr N p H y s r o p A T H o L o c y
tt
2tc
g
o
t
o /to
I
r
L
a@
c
(J
o
o
o
0
a
F l c . 1 5 2 . T h e i n c u b a t i o no f c i t r a t e db l o o d a t 3 7 " C c h a n g e so n l y l i t t l e t h e a m o u n t o f
c h o l e s t e r o li n C a s e ( a ) , w h i l e i t d e t e r m i n e sa r a p i d d e c r e a s ei n C a s e ( b ) .
THERAPEUTTC APPROACH
455
>e
bl
L
c,
E
L
o
a
E
I
E
o
L
o
E
u
o
U
# of rashinos
f of rashinqs
F t c . 1 5 3 . T h e w a s h i n g o f h u m a n r c d c c l l s w i t h s a l i n e i n d u c e sa v e r y s l o w d e c r e a s e
i n t h e i r c o n t e n t i n c h o l e s t e r o li n C a s e ( c ) . T h e h e m o l y s i ss t a r t s i n t h i s c a s e a t t h e
lOth washing. In Case (d), such an effcct appears after the second washing.
456 /
RESEARcHtN pHysropArHoLocy
THERAPEUTTc APPROACH
457
a n o f f b a l a n c eA. h i g h c c l l r l a r . w i l h a r e l a t i v e l yn o r m a l o r l o w p l a s m a t i c
cholesterol,correspondsthu:; to an abnormalcholcstcroloffbalanceprcd o m i n a n ta t t h e c c l l u l a r I c v s l , 1 1 ' h i 1ag r c l a t i v e l yl o w c e l l u l a r a n d h i g h
plasmaticcholesterol,
to thc opposireoffbalance.
By now considering
cholesterol in its anti-fattyacid rolc, thcscchangcspermit further interpretation.
As we haveseenabovc,cholestcrolcorresponds
ratherto the anti-fatty
acid constituentscontrollingthc normal polyunsaturated
fatty acids and
thus differsfrom the corticoids*hich representagentsopposingthe abnormal fatty acids.The ultimatccauseof hypercholesteremia
would logically
b e s o u g h ti n a q u a n t i t a t i v c layb n o r m a l l h
y i g ha m o u n to f q u a l i t a t i v e lnyo r m a l
fatty acidspresentat thc low'crlcvcls.
A l t h o u g hh y p c r c h o l c s t c r c n r ci ap r c s c n rbsy i t s e l fo n l y a r e s p o n s et o a
fatry acid offbalancctaking placcat lowcr levels,the high amount of sterol
presentin blood can cause,by itsclf,a scricsof disagreeable
manifestations.
Hypercholesteremia
can thus inducenoxiousmanifestations
although,teleologicallyspeaking,it is directedto correctan offbalanceat a lower level.
The effectof this chronicrichncssin blood cholesterolis manifestedin the
circulatorysystemand athcromatosis
appcarsas an immediateresult.The
precipitatingtendencyfor any ncw incrcascin bloodcholesterolexplainsthe
appearanceof crystalsin the cclls of the intima and secondarilythe appearanceof atheromas.
Therapeutic Attempts
This pathogenicconccptof artcriosclerosis
presented
above.has guided
. s c a r l y a s 1 9 4 2 ,w e u s e da c i d l i p i d i c f r a c t i o n s
o u r t h e r a p e u t iac p p r o a c hA
of organs in a tentativethcrapy to control hyperrensionand indirectly
with intcrestingimmcdiatercsults.We learned however,
arteriosclerosis,
that not only were thc effectstcmporarybut also that after sometime, the
fatty acidsinduceda progressive
administrationof unsaturated
increasein
t h e a m o u n to f c h o l c s t c r opl r c s e n ti n b l o o d .
as a secondaryblood responseto
The conceptof hypcrcholcsteremia
a cellular fatty acid predominancccxplaincdthis occurrenceand led us
also to the further developmentof this approach.With cholesterolas an
anti-fattyacid agentwhich appearsin high amountsin blood, as a response
to an excessof fatty acids at the cellularlevel,we tried to influencethis
secondaryresponseby acting on thc existingprintary cellular offbalance.
could be obtainedby supplyingsubstances
other
A decreaseof this response
than cholesterol,acting as anti-fattyacid agcnts.If the cellular fatty acid
offbalancecan be controlled,the organismas an entity would no longer
be obliged to respondto the occurringoffbalanceand manufactureblood
458
REsEARcHrN pHysropATHoLocy
THER^PEUTIC
APPROACH
459
460
R t , S E A R C H l N
P H Y S I O P A T H O L O c Y
C H A P T ETR 5
THERAPEUTICAPPROA(IH
TO CANCER
/\
o F T H Eu L T T M A TnEr u s o f o u r r e s e a r c h a s b c c n t o t r y t o u t i l i z e
L/*E
in cancerthe knowledgeobtainedfrom investigations
of the gencralprobIems of pathologyand therapy.Encouragedby the resultsof biologically
guided therapyin many other conditions,we have appliedit to the treatment of malignancy.
As we have mentionedbefore, differencesbetween animal cancers,
both experimentaland spontaneous,
and human cancersrepresentone reason why an agent,howevergood its resultsin animals,may not apply to
human malignancy.Another factor, conductof treatment,is no less important.The main characteristic
of our therapeuticapproachresidesin the
fact that treatmcntis continuouslyguidedby data rcprescntingthe actual
conditionof the subject.At leastfor the moment,it appearsimpossibleto
recognize,through suitabletests,the patternspresentin animalsso as to
apply them to guidcd therapy.Thcrefore,we have becn obligedto do our
therapeuticresearchin humans,reservinganimal studiesfor limited problems.This situationhad led us to emphasize,
always,the experimentalnature
o[ our thcrapeuticeflorts in humans.Although we startedwith desperate
terminal cases,frank immediatesubjectiveand objectivebenefits,even
though temporary,were obtainedfrequentlyenoughto encourageus to go
on. Together with the above mentionedconsiderations,
they seemedto
justify the continuationof therapeuticresearchin human patients.We will
try to reviewas objectivelyas possiblethe resultsobtainedwith therapeutic
methodsand agentsevolvedover the years.
In 1927, a 33-yearold woman with typical preterminalcancerof the
stomach came under our care. In the highly emaciatedpatient, a hard,
461
462
nEsEARcH tN pHysropATHoLoGy
irregular mass frlling the entire epigasterwas palpable. Radiological cxamination indicated a prepyloric gastric tumor. Laparotomy showed an
inoperabletumor of the stomach,with the omentumand lymphaticgJands
greatly involved and evidenceof multiple peritonealand liver metastases.
In view of the patient'sgeneralconditionand the fact that the pyloruswas
only partially obstructed,no surgicalprocedurewas performed other than
biopsy of one of the metastasesin the omentum. The biopsy showed an
adenocarcinoma
Grade III of gastricorigin. Treatmentwas not prescribcd.
I saw the paticnt two yearslater in apparentlygood health.clinical and
radiologicalexaminationat that time showedno tumor. The patient attested
to receivingno treatment.At thc time of the operation she had been trvo
months'pregnant.we had attributedher amenorrheaat that time to the
advancedcachccticcondition.She had given birth at terrn to a normal grl.
Hers was one of thosecasesusuallycatalogued
as "spontaneous
remission."
Since then, I have analyzedmany of the published observationsof
casesof so-callcd"spontaneousrcmission"of cancer alwaysto find a turnilg point that coincided with the interventionof some event usually considered to have no possiblesignificancefor malignancy. The fact that such
eventshavenot inducedsimilarchangesin other cancerpatientshas made
them seemunimportant to many investigators.
While not regardingthem as the only causeof favorable changes,we
havc not eliminatedthe possibititythat sucheventsmay have a contributory
role. We must recognizethat, if such events in themselvesappear to be
powerlessto changethe courseof cancer,they may intervenein conjunction
with, and potentiate,another factor also powerlessin itself to induce a
change.
It was with this concept in mind that we reviewed the case of the
woman with stomach cancer. We consideredthe possibleeffects of two
factorswhich apparentlyintervenedconcomitantly:pregnancyand surgery.
We then begana seriesof experiments.
Ehrlich mammarycarcinomawas graftedin two groupsof female mice,
onc pregnantand the other not. In eachgroup,half of the mice were kept
as controlswhile the other half was submittedto a sham surgicalprocedure
consistingof a laparotomy in which multiple ligatures were performed.
Growth of the tumors and survival times were noted. Compared with nonoperated,nonprcgnantmice servingas controls,both the pregnantmice
and the surgicallytreatedmice showeda slowingdown h the evolution of
cancer lesions.It was in the group of mice, both pregnant and surgically
treated,that a temporary arrest in tumor evolution was seen.In some ani-
:;tiitl)r,l:.i
liti,i
'
i,,I'{,tr,l
THERAPEUTIC
APPROACH TO
cANcER
463
'n
malseventemporaryregression
was noted; 1/20 the tumor regressed
entirely.
PlacentaExtracts
Theseexperimentsled us to try an extractobtainedfrom placentaautolysateswhich, in our opinion at the time, would reproduce,up to a point,
some of the conditions present in these experiments.Human placentas
were autolyzedby being maintainedfrom severalhours to a few days at
37" C, and an alcoholicextractwas obtained.The alcoholwas eliminated
tfuough distillation in vacuum. The residueproved non-toxic in animals
and was injectedintramuscularlyin somc terminal cancercases.
Imprcssiveresultswere observedin the first cases.Pain was markedly
diminishedand, in someinstances,
disappeared
entirely.objective changes
in the tumors could be noted. while only temporary resultswere seenfor
most of thesecases,for some the resultsappearedto last a long time. Their
number would excludepure coincidence.
Mr. H. 8., 56 yearsold, came under our care with a cancerinvolving
more than half of the right parr of the tongue.Multiple large submaxillar
and cervical gland metastaseswere present,two of them being approximately 8 cm. in diameter.The mouth lesion was very painful and bled
occasionally;there was moderate pain in the ganglionar metastases.A
biopsy performed at a much earlier stagehad shown a squamouscarcinoma. Consideredinoperable,the subjecthad not receivedany treatment
except for pain palliation.
We administereddaily intramuscularinjectionsof 5 cc. of the placenta
extract. Except for a limited local reaction at the site of injection, no disagreeableeffectswere seen.On the contrary, after each injection, the pain
in the tongue was reduced for a few hours. It disappearedentirely after
one week of treatment.During the secondweek of treatment,the tumor of
the tongue, as well as the metastases,
beganto decreasein size. The local
reaction at the site of the injectionsincreased,however,to such an extent
that we were obliged to stop treatmentafter 5 weeks.In spite of this, the
lesionscontinuedto decreaseso that the tongue tumor was no longer
palpableafter two months.At that time, the gland metastases
were reduced
to approximatelyone and a half centimetersin diameter.The patient's
general condition was much improved and he gained weight. In another
month, exceptfor a scar on the tongue,no other pathologycould be found.
We followed this casewithout treatmentfor anotheryear and a half during
which time there was no recurrence.After that, the patient left town and
$'e were unableto reestablish
contactwith him.
464
REsEARcH rN pHysropATHoLocy
Mrs. B. A., 44 yearsold, cl..:i u;r 'cr our care with a massivetumor
filling the entire vagina.The crnrl;:;rn had becn diagnosedg months previouslyas carcinomaof the ccr'..x "1;.:h had invadedthe parametriaand
was propagatingtoward thc .,ig.;- r. .', Liopsy made at that time indicated
squamouscarcinomaGradc III. .\: ll c patientwas consideredinoperable
and refusedany other treatment,only scdationwas prescribed.when we
examinedher, the tumor was pr-iilrdirg from the vaginaas a hard mass.
Rectal examinationrevealedin.,'asionof the entire recto-vaginalwall. The
patient received a daily injection cl the placenta extract preparation for
45 days, after which she intcrrupleJ the treatment.The pain had been
entirely controlled in less than a w'cck, but no other changeshad been
observed.
She returnedthree months latcr, having receivedno treatmentin the
interval. Examination revealed complcte disappearanceof the vaginal
tumor, with the cervix entirely rcplaccd by soft scar tissue. we followed
this casefor two years,during u,hich no further treatment was given and
the patient showedno recurrencc.
Mr. A. N., 40 years old had an extensivecancerof the cheek, with
massiveulcerationresultingin a largc communicationbetweenoral cavity
and exterior. Occasionallysmall hemorrhagicepisodeswere experienced.
The patient had had severalcoursesof radium therapy. when he came
under our care, he had multiple lesions,and severalbiopsiesperformed at
that time revealedactive carcinoma in atl lesionstested.The patient receivedintramuscularinjectionsof 5 cc. of the placentaextract preparation
daily for l7 days. A massivehemorrhageoccurredat this point, treatment
was stopped, and he went home without further medication. when the
patient returnedthree months later, scar tissuecoveredall areaswhere the
tumor had been seen previously.clinically no trace of tumor could be
found. In a few months,the patient'scondition was good enough to allow
his surgeonto attempt a skin graft to cover the big opening in the cheek.
This was not successful.The graft from the skin of the neck unfortunately
underwentnecrosis.
In severalother cases,similar subjectiveand objective changeswere
observedwith use of the same alcoholicextract of human placentaautolysates.During this time, we attemptedto substitutecow placenta,utilizing
both fetal and maternalparts, which are easily separablein the cow. In a
relatively small number of casesin which these products were used, we
could see no differencesin the influenceof placentaextract accordingto
origin. Poorer resultswere obtained with extractsusing fresh placentain-
465
..":i;"
:..:.rl;i
:j
l"'&:{lr.,l i rs.":r
466
nESEARcH rN pHysropATHoLooy
THER^pE,UTlCAppROACH To
CANCER
467
468
,i
R E S E A R C Hr N p r t y s r o p A T H o L o c y
Mrs. D. A., 68 yearsold, had a cancerof the left breastfor which she
had undergonea radical mastectomyfour years previously.pathological
examinationof the lesionhad shown an adenocarcinoma
Grade IV, with
g a n g J i o n ai n
r v o l v e m c n tW
. h e n t h e p a t i e n tc a m c u n d e r o u r c a r e s h e w a s
bedriddenwith a diagnosisof multiple bone metastases.
Radiologicalexaminationshowed multiple osteolyticlesionsin the pelvis, femur, lower
spine.ribs and skull. We institutedtreatmentwith cod liver oily fatty acids
i n g e l a t i n ec a p s u l e sT. h e d o s e w a s p r o g r e s s i v e il n
y c r e a s e db. y 0 . 2 5 g m .
rncrements,until it reached3 grams a day. ortho-phosphoricacid was
addedorally in dosesof vq cc. of a Sovo solutiongiven in water in order
to controlthe pain which appearedafter administration
of the capsulesand
wasof an alkalinepattern.Improvementbeganin a few daysand continued
so satisfactorilythat in lessthan six weeksthe patient was up and about.
Five monthslater, with bone lesionshealed,the patientwent home. I saw
h e r i n I 9 4 1, a l m o s tt w o a n d a h a l f y e a r sl a t e r ,d u r i n gw h i c h t i m e n o t r e a t m e n th a d b e e ng i v e n .w h e n e x a m i n c da t t h a t t i m e ,s h e a p p e a r e di n e x c e l lent conditionS
. u b s e q u e n t l by e
, cause
o f t h e w a r , I l o s t c o n t a c tw i t h h e r .
The increaseof pain, and especiallythe frcquent appearanceof pain
o [ a n a l k a l i n ep a t t e r na f t e r e x t e n d e dt r c a t m e n t c, o n s i d e r a b l lyi m i t e d t h c
use of thesecod livcr oil fatty acid preparations.
Furthermore,an incons i s t e n ciyn o b j e c t i v c h a n g ew
s a ss e e ne v e nw h c na d m i n i s t r a t i ow
n a sg u i d e d
b y t h e a c i d o r a l k a l i n ec h a r a c t eor f t h e p a i n , I n m o s t p a t i e n t s f, a v o r a b l e
objectivechangcswere only temporary.
During this research,wc observeda very favorablcrcsponscin some
casesof hemorrhage,
especiallyof the long-termoozingtype, treatedwith
thesepreparations.Bleedingusuallystoppedafter one injection of I cc.
of a l0olc solutionof unsaturatcdmembersof cod livcr oil fatty acids.We
still use this preparationfor this purpose,as mentionedpreviously.
Among thc group of lipids opposedto fatty acids,we first usedcholest e r o l w i t h t h e i n t e n t i o no f t r y i n g t o i n f l u e n c ep a i n h a v i n ga n a l k a l i n ep a t tern. The effect was much less impressivethan that obtained with fatty
a c i d si n p a i n o f a c i d p a t t e r n .I n s o m ec a s e so. b j e c t i v ec h a n g e sa l s o w e r e
o b s e r v e da l t h o u g ht h e y w c r e l c s s f r e q u e n ta n d l e s sp r o f o u n dt h a n t h o s e
secn with the fatty acid preparations.Cholesterolalone never produced
t o t a lc l i n i c a ld i s a p p e a r a n coef t u m o r s .
Acid Lipidic' Fractions and Unsuponifiable
The developmcntof the conceptof dualisticpathogenicintervention
o f t w o g r o u p so f l i p i d s l e d u s t o a t r e a t m e net m p l o y i n gt h e s et w o t y p e s
o f l i p i d c o n s t i t u e n tosf t h c b o d y c h o s e na c c o r d i n gt o t h e c h a r a c t e ro f t h e
T H E R A P N U T I C
A P P R O A C H T O
C A N C E R
469
manifestations.
We obtainedfrom differentorganicsourcesthe insaponifia b l ef r a c t i o na s w e l l a s t h e a c i d l i p i d f r a c t i o n H
. u m a n p l a c e n t aw a s w i d e l y
employed.The two preparations,acid lipid and insaponifiablefraction,
w e r e u s e d u n t i l 1 9 4 3 o n a b o u t 2 0 0 p a t i e n t s( i n F r a n c e ,a n d M e x i c o ) .
Somegood subjectiveand oblectiveresultswere observed.The subjective
changeswere most impressive.Frequently,an injectionof only I cc. of
the human placentaacid Iipid prepararion(5c/c in oil) controlledpain in
a few minutes,with relicf lastingfor hours or sometimeseven days. with
the samcpreparation,alkalinepain increased
after only a few minutesand
sometimesbecameunbearable.In casestreatcdwith placentaacid preparation, usingacid pain patternas a criterion,we observedsome significant
objectivechanges.
Mrs. B. B., 54 yearsold, with a papillaryadenocarcinoma
of the ovary
and multiple peritonealmetastases
found during exploratorylaparotomy,
.l
had rapidlyreproducingascites. hc patientrequircdrepeatedparacenteses
a t s h o r ti n t e r v a l sI.n t h c m o n t hj u s t p r i o r t o c o m i n gu n d e ro u r c a r e ,i t h a d
beennecessary
to tap her once a week or evenevery five days,Treatment
w i t h a c i d l i p i d s o f p l a c e n t aw a s i n s t i t u t e dw, i t h d a i l y i n j e c t i o n sf,i r s t o f I
c c . a n d t h e n o f 2 c c . o f t h e 5 v c o i l s o l u t i o n .I n l c s st h a n t w o w e e k s ,t h e
pain was controlledand much lcsslluid accumulated.
The patienthad two
more paracenteses
at two and three week intervals,after which fluid no
longerwas a problem.The multipletumor masses,which wcre very easily
felt throughthe skin after cach paracentesis,
were seento decrcaserapidly
and disappcared
in about two and a half monthsof treatment.After four
m o n t h s ,t r e a t m e n tw a s d i s c o n t i n u e a
d n d n o r e c u r r e n c ew a s s e e nd u r i n g
years
we
the three
followedthis patient.
W e s a w M r s . L . S . N . . 7 3 y e a r so l d , i n 1 9 4 2 i n a s u b c o m a t o usst a t e ,
w i t h d e e pj a u n d i c ea n d w i t h a h i s t o r yo f p r i m a r yt u m o r o f t h e s t o m a c ha n d
m u l t i p l e b i g m e t a s t a t i cl c s i o n so f t h e l i v e r . H e r c o n d i t i o n ,w h i c h h a d
s t a r t c da f e w m o n t h se a r l i e r w
, a sg c t t i n gr a p i d l yw o r s e .w h e n w e e x a m i n e d
h e r . t h e I i v e r w a s o c c u p y i n gt h e a b d o m e nu n t i l t h e p u b i s ,a n d p r a c t i c a l l y
c a c ho f t h e i n d i v i d u a m
l e t a s t a t itcu m o r sp r e s c n a
t t t h c s u r f a c co f t h e l i v e r
w a s e a s i l yp a l p a b l et h r o u g ht h c t h i n a b d o m i n awl a l l .
G u i d e db y t h e u r i n c a n l l y s e s - w i t h a h i g h p H a n d a l o w o x i r e d u c t i o n
i n d e x - w e s t a r t e dw i t h a t r c a t m e n w
t i t h l 0 % s o l u t i o ni n o i l o f a h u m a n
p l a c e n t al i p o a c i dp r e p a r a t i o nT. h c d o s e sw c r e i n c r e a s e d
a c c o r d i n gt o t h e
a n a l y s e su n t i l t h e y r e a c h e d3 i n j e c t i o n sd a i l y o f 2 c c . e a c h .T h e p a t i e n t
i m p r o v e d a, n d i n l e s st h a n u w e e ks h ew a s c o n s c i o u as g a i n .H e r c o n d i t i o n
c o n t i n u e dt o i m p r o v ef o r m o r e t h a n a m o n t h w h e n a r a p i d c h a n g ef o r t h e
\,\'orse
took place.As the analysesat this time showedthe oppositeoffbal-
470 ,/
R . E s E A R crHN p H y s r o p A T H o L o c y
ancepresent,the treatmentwas changed.In additionto infusionswith glucose and saline,glycerol in a dose of g drops daily with 20 drops of
coreminewas given.Again the response
was impressivcly
good.The general
conditionimprovedrapidlyand in lessthan a month, the patientwas out of
bed and startedto take care o[ her home. By this timc, the jaundicehad
almostcompletelydisappeared,
althoughthe liver remainedalmost of the
same dimension.The patient continuedto take glycerol for more than 6
months,lcadingan absolutelynormal life. It took almostone year for the
liver to come back to normal dimensions.
The patient remainedin perfect
healthfor the next 9 years.Severalcoronaryocclusions
led to her death at
the age of 83, from a myocardiarinfarction.She showedno objectiveor
subjectivesigns of recurrenceof either her stomachor liver condition.
Unfortunately,uncontrollablechangestoward rapid tumor gowth ultimately occurred in most casestreatcd with placentaacid lipids despite
favorableobjectivechangesat the beginningof treatment.
Clinlcaluseof the insaponifiable
fractionpreparationssupportedobservationsmade in animals,indicatingthe importanceof other factors for
obtainingfavorablechangesin tumors.With the insaponifiablefractions of
placenta,marked clinical effcctscould be obtaincdonly when the conclition
of the patient permittedthe treatmentto be continuedfor a long time.
Temporaryregressionor evenclinical disappearance
was effectedin several
cases,only to have the tumors start growing again,this time bcyond control
by the mcdication.Even at this point, it was obviousthat favorableinfluence with thesc two antagonisticgroups of lipids was dependenton using
the lipid which corresponded
to the patternpresent.It becameincreasingly
evidentthat changesin the patternoccurredduring treatment.Administration of acid lipidic and insaponifiable
fractionsinduced unfavorableresponses
in patientswith tumorsin which a predominance
of the samelipids
was indicatedby analyses.Pain increasedand unfavorablechangesoccurredin the evolutionof the discase.For cxamplc:
Mrs. A. D., a 42-year-oldwoman,operatedon for an adenocarcinoma
of the breast l8 monthsprior to comingunder our care, presenteda few
skin lesionsnear the operationscar.Radiologicalstudy revealeda few small
osteolyticlesions in two ribs, and one in the skull. The analytical data
showedlow urinary pH, high specificgravity and high chloride index, indicatingan offbalancewhich we attributedto predominanceof fatty acids.
Treatmentwith 2 injectionsdaily of i cc. of a 5o/ooily solution of the insaponifiable
fraction of placentawas started.This was later increasedto
2 cc. twice a day. The patientleft the city for a summervacationand took
the medicationwith her. Disregardingour instructionsthat treatment must
4 71
All
1 t L
R E S E A R C H
I N
P H Y S I O P A T H O T - O C Y
c o n t r o l l e da g a i nb y t h e s a m ep r e p a r a t i o nI .n a s m a l l n u m b e ro f p a t i e n t s ,
about 3Vo of the group of 200 treatedwith thesepreparations.
the favorableresultscould be maintainedover a numbcrof years
Even with the relativelystrict guidanceof therapy by the analyses
availableat the time, resultswere not alwaysfavorable.We attributedthis
both to the agentsused for therapyand the criteria employedfor recognition of offbalanccs.
For a long time, researchwas devotedto developing
meansto permit betterrecognitionof offbalances
and to ascertainthe value
of the variousanalysesusedas criteriafor the conductof treatment.Each
new urine or blood tcst was investigatcdas a criterion for the group of
lipids to be administcred.This lcd to bettcr resulrsin controlling pain,
i m p r o v i n gt h e g e n e r a cl o n d i t i o na n d e v e n i n o b j e c t i v cc h a n g e si n t u m o r s .
H o w e v e r ,t h e t e m p o r a r yc h a r a c t c ro f t h e c f T e c t o
s b t a i n e dw i t h l i p i d s
derivedfrom nornralorgansappearcdmore and nrorc evidentin long-term
appraisalof results.We changcd from human placenta to other sources
for both acid and insaponifiablc
fractions.We prcparedand used lipids
from differentorS,ansof cow, pig, tish. and chicken.We also used molI u s k s ,c h i c k e ne m b r y o s ,m o l d s a n d c v e n m i c r o b e s a. s w e l l a s m i l k a n d
eggsas sourcefor thcselipids.In one group of invcstigations,
we even tried
to use lipids of the organ from which the tumor derived.Most of these
preparationssatisfactorily
controlledpain, and in somecases,good results
wereseenin the growth of tumors.
T h e f o l l o w i n go b s e r v a t i o cno n c c r n sa c a s et r e a t e dw i t h t h e l i p o a c i d s
of humanblood.
A . M . , a 5 6 - y e a r - o l dn r a n .w a s r e f e r r e dt o u s b y h i s p h y s i c i a nw i t h a
diagnosisof cancerof the rcctum.Difficultyin defecation,mucosanguinolent discharges,and pain in thc rectal region had been increasingin the
t h r e em o n t h sp r i o r t o t h e d i a g n o s i sE. x a m i n a t i o nh a d r e v e a l e da t u m o r o f
a cauliflowertype, startingat about 4 cm. from the anal orifice and almost
entirelyfilling the rcctal anrpulla.A biopsyhad shown it to be an adenocarcinoma,Crade III. The patient had refusedsurgicalinterventionbecause,yearsbefore,a minor operationon his right hand had led to local
infectionfollowedby amputationof the hand.
At the time he was referredto us. his main complaintswere pain in
the rectum,radiatingto the left lc'g,and tenesmuswith frequentmucosanguinolentdischarges.
We employeddaily injectionsof I cc. of a 5 % olly
solutionof the acid lipidic fructionobtainedfrom human blood. The treatm e n t w a s c o n t i n u e df o r s i x w e c k s ,t h e i n j e c t i o nb e i n gg i v e n d a i l y d u r i n g
the first two weeksand twice a week thercafter.After one week. the pain
and tenesmusdisappcarcdand thcre was a decrease
in the mucosanguinol-
ro
cANCE.R
473
ent discharge.
There was also an obviousdecrease
in the sizeof the tumor.
In lessthan a month the tumor regressed
to one-fourthits originalsize;in
six weeks,digital and proctoscopic
examinationshowedno clinical tumor.
A whitish scar could be observedin the posteriorwall of the rectum.
Thereafter,the condition of the paticnt was followed indirectly through
reportsfrom his physician.Therc was no tumor recurrencein spite of the
fact that he receivedno further treatment.He died six vearslater from an
acuteparatyphoidinfection.
Our clinicalexperienceprovidedamplecvidenccthat the preparations
rich in polyunsaturated
fatty acids would influencepain as well as the
growth and evolutionof human cancers.Using the sameamountsof polyethenicfatty acidspreparedfrom varioussources,no differencesin efiects
could be noted. The effcct upon tumors in all instanceswas relatively
limited.
our next effort was to try fatty acidsunlike thosefound in the organism. They includednorbixine-the monomcthylester of the bicarboxylic
acid, bixine-which we uscd in a group of patientsbetween 1938 and
1940.with urinary specificgravityand pH as crireria,daily dosesof from
I mgr. to 100 mgr. were administered
to 30 preterminaland terminalpatients with pattcrnscorrespondingto prcdominanceof sterols.Even with
small doses,the changestoward a predominanccof fatty acids were impressive.Superficial,massivetumors were often seento melt away within
a f e w d a y s ,u s u a l l yl e a v i n gu l c e r a t i o ni n t h e i r p l a c e . - f h i sr a p i d c h a n g eo f
a m a s s i v ct u m o r i n t o a n u l c e r a t e o
d n e ,h o w e v e ru, s u a l l yw a s f o l l o w e db y a
manifestdeteriorationof thc generalcondition. With this preparation,
once the offbalancewas changcdfrom thc original to the oppositetype,
attemptsto control the new offbalance*'ere usuallyunsuccessful.
This led
us to discontinue
i t s u s e a t t h i s t i m e i n s p i t co f t h e r a p i d a n d i n t e n s i v e
c h a n g eist i n d u c e di n t u n r o r s .
Croups ol Agents
Parallelto theseresearchcs
on fatty acids,the usc of agentswith positive polar groups also underwentchanges.As previouslymentioned,it
appearedincreasing.ly
clear that no singleagentcould be effectivein itself
if an offbalancecorrespondine
to predominance
of fatty acidswas present.
use
Quite early in this researchit could be observcdthat the simultaneous
of two agentsfrom the samegroup appearcdto be a bctter procedurethan
u s i n ge i t h e ra g e n ta l o n e .I n a l i m i t e dn u m b e ro f p a t i e n t st r e a t e dw i t h s i n g ) e
agents,without impressivcclinical changes.bctter effectswere obtained
when combinationswere tried. Although it was difficult to ascertainin
474
R E S E A R c Hr N p H y s r o p A T H o L o c y
THER^PEUTIC APPRO^CH TO
C^NCER
475
476
RESEARCfI IN
P}IYSIOPATHOLOCY
s
#
F l c . l - q 5 . , , \ r t e r i o p ( ) \ t e r i t r r v i s r r . o f t h c c h c r t o f p : r ti c n t 5 m o n t h s l n t e r . w i t h t h e
y
i r p p f a r a n c c o f t h e n e r v b o n c s r e p l a r ; i n g t h c p i l r t o f t h c r i h r p r c v i e i u s ldcstroyetl.
T H E R A p L - L i T t cA p p R O A C H T o
c A N C E R
4 7 7
of the 6th and 9th left ribs. (F'ig. 154)The analysesshowedan offbalance
t y p e D a n d c o n s e q u e n t lsyh e w a s t r e a t e dw i t h b u t a n o la n d g l y c e r o l .T h e
patientshowedrapid changcsnot only in her subjectivefeeling,but also
o b j e c t i v e l yF.i g . 1 5 5 s h o w st h e r i b o s t e o l y t ilce s i o n sh e a l e da n d t h e m i s s i n g
bonereplacedby new bone.
In general,however,in spiteof somefavorablecases,the resultswere
not satisfactory,Even in patientsin whom tumors regressedand disappeared,recurrences
wereseenanywherefrom a few monthsto severalyears
after discontinuation
of treatment.By prolongingtreatmentover a period
o f y e a r s ,t h e p e r i o d o f i m p r o v e n l e nwt a s l e n g t h e n e idn s o m e c a s e s S
. till,
all too often the responseto treatmentwas only temporary.
Mercaptans
Other substances
with lipoidiccharacterbut with a polar group different from thc carboxyl were tested.Mercaptans,as lipoids with a thiolic
negativepolar group, were used.In 1942. we startedto study the therapeuticeftectof ethyl mercaptaninjectedintramuscularly,
in a l0o/osolution
in oil. (326)
Use of ethyl mercaptanhad to be limited to subjectswith an offbalancc
corresponding
to a predominance
of stcrols.Although only a few patients
were treatedbecauseof the offcnsiveodor, thc resultswere satisfactoryas
indicatedby the followingcasehistories.
F . C . - T h i s p a t i e n t ,a t t h e a g eo f 6 6 , h a d a s m a l lc a r c i n o m ao f t h e t i p
o f t h e t o n g u er e s e c t e a
d t C o l u m b i aP r e s b y t e r i aHno s p i t a lo n M a y 6 , 1 9 4 1 .
At the same time, a secondcarcinomaof the left lateral bordcr of the
t o n g u e w a s f o u n d i n f i l t r a t i n gi n t o t h e d e e p e rp o r t i o n s .T h i s l e s i o nw a s
t r e a t e dw i t h r a d i u mn c e d l e w
s i t h a t o t a ld o s eo f 2 5 0 0 m g . h o u r s .I n S e p t e n r b e r 1 9 4 1 ,a p r o p h y l a c t i cl e f t r a d i c a ln c c k d i s s e c t i o w
. hc
n a s p c r f o r m e dT
patient was well for 8 months thereafter,and then began to experience
sorenessin the regionof the scar on the lateralside of thc tongue.
I n A u g u s t 1 9 4 2 , l 6 m o n t h sa f t e r t h e t u m o r h a d b e e n i r r a d i a t e dt,h e
p a t i e n tc a m e u n d c r o u r c a r e . A t t h c j u n c t u r eo f t h c a n t e r i o ra n d m i d d l c
t h i r d o f t h e l e f t l a t e r a lb o r d e r o f t h e t o n g u e ,t h e r c w a s a n i n d u r a t i o no f
2 / l c m . i n d i m e n s i o nw i t h a n u l c c r a t i o no f 1 / 0 . , 5c m . c o v e r c db y n e c r o t i c
tissue.The induratedbasewas especially
developedanteriorlyto the ulcerated lesion.There was no evidenceof recurrcnceat the scar of the tip of
the tonguenor at the left sideof the neck.The latterlesionwas very tender.
A W a s s e r m a nann a l y s i sw a s n e g a t i v eB. i o p s yo f t h e e d g eo f t h e u l c e r a t e d
s q u a m o u cs e l l e p i t h e l i o m a .
tumorrevealed
T h e p a t i e n tw a s t r e a t e dw i t h p l a c e n t aal c i d l i p i d f r a c t i o nf o r t h e f i r s t
478 /
REsEARcH lN
pHysropATHoLocy
THERApEUTTC AppROACH To
CANCER
479
F I c . l - J 6 . C y s t o g r a mo f p a t i e n tH , A , b e f o r et r e a t r n e n l .
4lt0
R E S I J A R C } I I N
P } I } ' S I O P A T I I O L O C Y
t . l L , . 1 5 7 . ( y r t o g r a n r o f p a t i c n t H . A . . a f t c r t r c a t n r c n tu ' i t l r e t h y ' l n l c r c l p t u n .
r H E R A p E U r r cA p p R o A c HT o c A N C E R
4gl
482
RESEARcH rN pHysropATHoLocy
rary.
The use of sulfurized oil in conjunction with sodium thiosulfate has
beentestedin a sufficientnumberof cases(more than 75) to enableus to
recognizethat the combinationproduceschangesin pain and systemic
analysis,as well as reductionin the sizeof tumors,especiallywhen given in
adequateamountsover a long enougbperiod of time. In severalcases,
tumors disappearedfor many yearsfollowing this treatment.The following
are illustrativecases:
G. M.-In March 1944, this patient had an ulcerationof the cervix.
Biopsyshowedsquamouscell epithelioma,
Grade III. A total hysterectomy
was performed.She was treatedwith 3,600 mg. hours of radium in and
aroundthe ccrvix in April 1944.Five monthslater, there was evidenceof
local recurrenceand the patient was given 1800 r. of deep X-ray therapy.
In July 1945,examinationrevealedno evidenceof disease.
On October28,
1945,the patient was examinedat the Scott and White clinic in Temple,
Texas, and a diagnosisof extensivemetastaticcarcinoma of the para-
493
metria was made and further deep X-ray therapywas advised.The patient
refused this.
She cameunder our care in November1945.She was extremelyweak
and showedevidenceof considerableweight loss, weighingonly 86 lbs.
Hematuriaand dysuriawere the principalcomplaints.Multiple largetumor
masseswere palpablein the pelvisand extendedinto the abdomenabove
the umbilicus. The largest mass palpable,about the size of a big grapefruit, was in the right lower quadrant.The tumor was found to have invaded the bladder, too.
The treatmentwith which we started,t.i.d. intramuscularinjectionsof
I cc. of placentafatty acids, Ljvo in oil, was changedafter one week, in
view of the preterminalconditionof the patient.A hydropersulfide
preparation containingI o/csulfur,and sodiumthiosulfatelOTo wasadministered
orally. we startedwith a doseof lq cc.threetimesdaily of hydropersulfide
and Yz cc. thiosulfateand increasedit progressively,with ameliorationof
the generalconditionand disappearance
of the hcmaturia.After a month,
the dosagereached3 cc. of the first and 9 cc. of the latterpreparationdaily.
Under this treatment,the patientcontinuouslygainedstrenglhand weight.
The hematuriadid not reappear.
In May 1946,she was admittedto the Universityof Chicagoclinic. A
large fixed, firm, irregular tumor masswas still presentin the lower abdomen risingfrom the pelvisto the umbilicus.Althoughher generalcondition
had improved,the patient againhad urinary frequencyand urgency,and
cystoscopyrevealeda severecystitisand severalsmall stonesin the bladder.
An intravenouspyelogramshoweda right hydro-nephrosrs.
The treatmentwith hydropersulfide
and sodium thiosulfatewas continued for 4 months in Chicagoduring which time the abdominalniass
becamesmaller,softer, less fixed and was no longer tender. By August
1946, her weight was 136 lbs., a gain of 50 lbs. sincestart of treatment.
The treatmentwas continuedduring 1947, althoughabdominalexamination did not revealany palpablemasses.On rectalexamination,however,
the pelvis appearedto be frozen but no definitemass was felt. Cystoscopy
showedseverecystitis,bladder calculi and a distortedbladder.Although
shepassedseveralstonesand gravel,her urinarysymptomspersisted.
In December1948,an attemptwas madein Texasto removea bladder
after lithotrity.The bladderwas perforatedduring
calculustransuretherally
this procedureand a recto-vasicalfistularesulted.The patient'slocal physician in Texas believedthat the patient was terminal, but we insistedupon
This was done, and the surgeonreportedthat there was no evidenceof any
a colostomy to divert the fecal stream as an immediate first procedurc.
{84
R E S h A R (
l N
P H Y s l ( ) P d T H O t . ( } C t ,
'fhe
pelvic clr abdominal nrasses.
patient made a slow rccovery.According
to rcportsto this date. l6 ycarsaftcr start of treatntent,no recurrencehas
beennoted. Recentlythc colostomywits closed.the patientbeing in gtxxl
condition.
M r s . M . L . - l n N o v e m b e r1 9 4 1 ,a t t h c a g e o f 4 0 . t h e p a r i e n rh a d i r
lcft oophorectontyfor a multi-loculatedovarian tumor with ascitesand
peritonealinrplants.Thc pathologicalfinding was papillary cyst adenocarcinoma of the left ovary. Without any othcr treatnrent,she rentainedfree
Flc. l.5tt, Photonticrograph of l;-nrph node fronr ('ase [:.H. showing metastatic adenoc a r c i n o m aC r l l t ( 4 o O r ) .
T H E R A p L U ' r l c A p p R O A C r rT u c A N C C . n /
495
:$
r1ffi
i$
's
s,x
:*
TF
"J
2 2/ 4 8
l ' t t ; . l - 5 9 . A n t e r o p o s l e r i o rv i c w o f c h e s t i n ( ' i l s e F . . H . a t c o n c l u s i o no f l c s l o s t e r g n e
l . n d d c e p X - r a y t h c r a p ! 't o \ p i n c . s h o w i n gd c r t r u c t i o no f n r e d i a l ' . r o f l e f t c l a v i c l e
r n d m e t a r t a t i cr i h l e . ' i o n r .
486
R E S I ] A R C H
I N
P H Y S I O P A ' IH O L O C Y
tQ/ 24/48
F t c . 1 6 0 . A n t e r o p o s t c r r ovr i c u ' o f c h c s l i n C ' a s c L . H . t * ' o n r o n t h s a f t c r r t o p p i n g
t e s t o s t e r o n et n d X - r a 1 ' t h e r a p y . s h o w i n g f u r t h e r i n v o l v e m c n t o f m e d i a l r . . r o f l c f t
clavicle.
T H I R A P I T U T I C
A P I T R O A C i I T O
( ' A N C L R
4ti7
F t u . l 6 l . L a t e r i r lv i c w o f t h o r a c i cs p i n ei n ( l a s c F - . H .a t t i r r r eo f : r d m i s s i o ns.h o w i n g
e r l e n s i v e m e t a s t a t i ci n v o l v e m e n lo f v c r l e b r a ew i t h c o l l a n r c o f l i f t h a n d n i n t h v e r t e brae.
413tt
R E S E A R C } TI N
PHYSIOPATHOI.-OGY
twelve week period gave complete pain relief. During this time, the patient's generalcondition improved rapidly, Motion and sensationreturned
to the toes,feet and finally to the wholc leg. By April, the patient was able
to sit clutof bed, By the end of May, shc was ambulatorywithout requiring
a braceor cane.Hcr only complaintwas a mild facial acnethat developed
durins the courseof treatment.
I'llLRAPlll".itt(
At't,R()iclt
I () l ANCI:R
.lse
F l o . 1 6 . { . [ - t t e r a l r i c u o f t h o r . c r c : p i n c i n ( ' . r r e l ' l l . f o u r r r . n . h r a f t c r h c g r n n i n gt r f
in ull reltchr;re.
trealment, shetwing considelalrlc bone 1c*a..tr,r.t
490
R F . S E A R C HI N
PHYSIOPATHOLOGY
F ' t c . 1 6 , 5 .A n t e r o p o s l c r i o rv i e r + o f l ef t s h o u l d er r eg i o n i n C l s c E . I l . f o u r n r t i r r t h r
a f t e r b e g i n n i n go f t h er a p y . r l r o * i n g c o n s i d c r a h l cr e p a i r o f l e f t c l a v i c u l a r a n r l r r h
lesions.
t H L RA P l: Lj't l(:
A P P R0AClt
TO
CANC[,R
491
HvdrcrnaphtlralencPersulfides
good resultsobtaincdwith ntercaptans
Thc exceptionallv
in trcatingcanccr
in humanscould only partiallybe rcproducedby thc differentagentswith
bivalentsulfur in the polar group we tested.In addirionro the hydropersulfidesof the falty acids,we tricd to prepareand study other groups of
persulfidus.Thc great ability of the products of hydrogcnizationof
( tetralin) and dccahydronaphthalene.such as tctrahydronaphthalcnc
(decalin)
to
fix
peroxides,
oxygenas
naphthalene
led us to try to fix sulfur
on their mr:leculcs.
Thescsubstances
wcre trcatedwith sulfur
as pcrsulfides
i n c o n d i t i o n s i m i l a rt o t h o s cw h i c h l c d t o f i x a t i o no f s u l f u r t o f a t t y a c i d s .
The agentsobtained through this fixation on tetralin wcre particularly
studied.
We havementionedthe pharmacological
studyof theseproducts.Basr-'d
on their biological action. they appearedto bc intcrnrediarybctweenmercaptansand fatty acids pcrsulfides,with the big aclvantageof having a
bearableodor. In humans.we utilizedthis producein casesof rvhich.most
are still under obscrvation.In gencral,the rcsultsobtained were good.
492 /
R E S E A R c Ht N p H y s r o p A T H o L o c y
THl:RAt,EliTtc
Af l'l{()A('l{ 1 (1 (.AN(,tR
491
Butanol, Glycerri
During the periotj when mercaptansand other sulfur-containingagents
were being studied.attentionalso was ccntcredon butanol in the group of
anti-fatty acid agcnts.while butanol'seffecr upon pain and other subjective manifest:rtionsappearedevident from thc beginning of its use, the
inffucnceuFJn tumors seemed.snrall.'fogelher with glycerol, however, it
producedsevcrallong-lastingobjectivcchanges.Characteristically,
in nost
of thesecasesrecurrenccsapgraredonly after ntany yearsof normal activc
life during which therc was no clinical ntanifestation
of cancer.In some
cases,howevcr.therc werc no recurrcnccs.
l - l a . 1 6 7 . f v l y e l o g r a mo f p a t i c n t \ 1 . H . r h o w i n g t h e l l o w o f l i p o i d o l a r r e s r e da t t h e
ievel of {-T.
.{9.1
R[,SEARCH IN
PHYSIOPATHOLOCY
'l'he
I - ' r t ; .l 6 t { . l Ul c l r t g r a r n o f t h c p l t i c n t t r | . H . , r h o i ri n g t h e c o n r p l c l c d i s a p p c a r a n c ct r {
thr obstruction.
Ill[:RAPLUII('AP]'ROACtt
T() CANCER
4q.5
par&lysis.
In 1943.anotherrecurrcncew&strcatedsurgically.ln Junc 1945,
the same procedure was repcated.Immediately after thc last operation,
there was still another recurrenceand the tumor this time started to grow
rapidly. Severepain was only stighrlyrelievedby narcotics.Radiothcrapy
was refused,in spiteof the massivetumor and pain.
Thc patientcanrc under our care in Decernber1945. u,ith scveraltumors occupyingthe right parotid region and cxtcndingbckrw thc nrirndibula. He was using variousnarcoticsr.r,rth
lirtlc cllccr. Wirh the urinary
chlorideretcntionindcx, pH and specificgravityls critcria.trcatnlentwith
l - t t ; . 1 6 9 . P l t t i c n t l . l l . w i t h a r c c u r r c n tc h o n u r o n t v r r l s i i r r ( ) t o
r lfl t h c r i g h t p i r r t l t i r , l
g l a n d b e f o r c ; t n d a f t c r t r c a i n t c n l . - l ' h es c a r i r f r o n r r c p c a t c dp r e v i o r r s u r g i c i r il n t c r ,
v c n ti o n s .
s o d i u mt h i o s u l f a t a
c nd hydropcrsulfidw
e sa s s t a r t c d .t . n d c r t h i s t r c a t n r c n t ,
which lasteda rveck,the lesionappcaredto be unfavorablvinfluenccdand
pain increascd.With the urine chlorideindex usedas u critcrion.the treatnrent was changed.I cc. of butanol 6.,5r;lwas administercdurally threc
timesa day alongwith 0.3 cc. of glyccrol.After 4 da;-s,the doscof butanol
was incrcasedto 2 cc. three timcs a day. Pain was rclisvedin a fcrv days.
Rapid disappearancc
of the tunror masscsfoliowed.The same trcatnrcnt
rvascontinuedfor one year. Sincethen. the paticnt has been u,cll and is
cnjoying good health without any rccurrcnccas of this date. {F'ig. l69 )
E . M . - l n 1 9 3 . 5a, t t h e l g e o f 4 2 , t h i s p a r i e n rh a d a r i g h r r a d i c a ln r u s tectomyfor adenocarcinorna
of the brcast.ln I 940. recurrentnoduiesappearedin the linc of thc scar. One of thesewas bit-rpsied
and showeda
recurrent adenercarcinoma.
Cradc IIl. The pirticnt wAS then treated
with deep X-ray therapy,3800 r, beingdeliveredthroughfive {ieldsto the
496 /
R E , s E A R c Hr N p H y s t o p A T H o L o c y
right chest and axillary regions.A daily dose of 200 r. was given for nincteen days betweenDecenrber1940 and January 19.11,using tbe following
f a c t o r s :2 0 0 k v , . 2 5 m a . , 5 0 c m . , l , i m m . C u l n d I m m . A l f i l t e r . I n
May 1941, the wound arca and recurrentnodules(/.'is,.170) were excised
and a skin graft was used to repair the defect.
In July 1943, skeletalmetastases.
predominantlyosteolytic in nature,
were reported in the fourth, fifth and twelfth thoracic vertabraeand first,
second,fourth and fifth lumbar vertebrae,the first sacral segmentand the
l c f t ; r l l ro f t h e \ i r c r u n t l. l t e r c r r l s r t l . r 'i n r o l l c r n c ntlr f 1 [ g o u t e rp o r t i t t n< , f
t h c l t f t i i i t i t t tr r r l r -t il t e i r r n e rP t r r t i o nt , i t h c r i g h t i l i u n r n e l r t h e s a c r o i i i a c
'l
i o i n t . h c l ; r t r u n lr c c c i r e r rl r : u c o n cul o u r s ct r l d c e p\ , r a l t h c r a p yo v e r t h c
\ p i t ) ci t n d I ) ( ) s t c r i (I ) rc l rr r . t i r r 'l c t ; r ld , r " ch c i n ul g o ( ) r . * i t h t h e s a m cf a c -l
I t r r : . I j o l l c r r i r r tt h r s .h t ' r 'r ] r t ' n r t 'cr , ; l r r c r i . h c | ; 1 i n i n h c r h u c k . w h i c h h a d
c t t n l i n c t ' ll t f t l ' , r l r c t i . x l r r c o t t r r t l c n r l ' r l lr r : l i r - ' r e tlli n r . lr l r c b e c a n l c a n r b u l i l t e r r t ' .
I l r J l r t t r t ; r r r l ( l ' 1I l t o l t r .r . l . p r i r r r l r r r , . r1r ' r u l r c r l . t t l r i t h c p a t i e n t r v a s
; t g l t i t l e , r l t i t n r ' r "kl \ h r ' L Jl t l ( ) s l r r l t h r t t n t c \
a=
.t'r'
4lts{t-$.-
#r
=. ..- ELtil
-'
-{ f --i 1rfi- E
-of,:ryi
'
' fuf
,--f',
-t
_-.z _- !
d$$i${
-*.
rn*-hl1*
"
fu$ e**t:l: " "'
*t
*i*;.
*'&
{F
i*.j
wr*"f-;
'':i'
._;ffiSp$
*
tuoI
*.-x,*
l - t c . 1 7 0 . P h o t r . r n r i c r r r g r a p hr r f r e c u r r c n t r L i n n t r t l u l c i n ( ' l r e
c a r c i n o n t a .C r l l l ( 4 0 0 x ) .
T H [ . R A p F . L r T t ( ' A p p R ( ] ^ ( ' H1 ( ] c A N C E R
497
especially the frrst, fourth and lifth bodics and transverseprocesses.De;xlsitswere presentthroughoutthe sacrum,in txlth iliac, the right ischiurn,
left acetabulumand upper fcmurs. Another seriesof deep x-ray treatments rvas uscd on the same areas of thc spinc and pxrsteriorpelvis as
before,the total dose this time being2000 r., usingthc samefactclrs.There
F r c . l 7 l . A n t e r o p o s l e r r o rv i e w s
of thoracic spine in Case E.M.
a l t i m c r . r f a d m i s s i o n ,s h o r v i n g
m o t a s t a t i ci n v o l v e n r e not f v e r t c brae.
F t r ; . I 1 2 . 1 . . ; rrtuel v i c w 6 f t h e
t h o r a c i cs p i n e i n ( a r e E . l v { .a t
tirnc of irtlnrisrion. showing
n r c t a s t i - t t iicn r , o l v c n r e n to f v c r tcbrac.
4q8
R I " : S t : A R ( : HI N
PllYSl{)PATllOt,ocY
the riglrt chcst region were well healedand therc was no evidenceof local
recurrenccof the tumor. I'here was no superficiallymphadenopa$y.The
liver area was tender to pressureon dcep inspirationbut the liver was not
palpable.There was tcnderncssover most of the vertebraewith pressure.
Patellar rellcxeswere hyperactivebilatcrllly. X-rays showed widespnead
skeletalmctastases
involvingthe dorsaland lumbar vertcbrae,the sacrum
a n d t h c p e l v i s .T h c l c s i o n sw c r e p r c d o m i n a n t l yo s t c o p l a s t i c( F
. igs.l7l,
1 7 2 .I 7 - t)
THER^PEUTIC ^PPRO^CH TO
C^NCER
499
acid. After five months of treatment,the urine became alkaline and remained so until the patient was dischargedfrom the hospital.(FiS. 174)
Clinically, pain was completely relieved after about one month. The
patient's appetite improved and she began to gain weight and to feel
stronger.After five months, she was mobilized with a brace and showed
good progressin the ability to walk.
No changeswere observedin monthly X-ray studiesduring the first
four monthsof treatment.In the X-rays taken in July, the fifth month, at
the time of urinary pH change,a few lesionsbegan to show areasof decreaseddensity.These changesprogressedfairly rapidly thereafter.
zura
aa
co
7a
r2
6C
6'
al
30
56
5l
aC
aa
.,O
Ftc.2
F t c . 1 7 4 . p H o f d a i l y m o r n i n g u r i n e s p e c i m e n si n C a s e E . M . d e t e r m i n e dc o l o r i m e t r i c a l l y ,s h o w i n gc h a n g e sf r o m a c i d t o a l k a l i n e5 m o n t h s a f t e r t h e b e g i n n i n go f t h e
treatment.
-500
'
Rr.sEARCt{ tN
pHysl()t,A'rHot.()cy
abdominalpain. An cxplclratoryoperationrt-'vealcd
nn obstructioncaused
by a tumor of the bowel that had spreadto the lymph nodesthroughoutthe
abdomen.A by-passingoperationwils performedto rclieve the obstruction,
A picceof the massand someof thc involvedlymph nodes were removed.
The pathologicaldiagnosiswa.sfibro-sarcoma.
F r c . l 7 - 5 .A n t e r o p o s t e r i o rv i e w s
o f t h o r a c r cs p i n e i n C a s e E . N l.
l i f t l ' - s c l c nn r o n t h si r f l c r o n s e to f
thcrapl', showing almclstcomp l c t c r er t i t u t i o n o f n o r m a l h o n e
5 tr u c tu r c .
Fri;. 176. I-ateral view of thoracic spine in (-ase E.M., fifi1'\cvcn rtronlhs lftcr onsct of
therapl. shou'ing almosl cttnr.
p l e l e r e s l i t r r t i o no f n o r m a l b o n e
\ l r U C t rUC .
On lvtarch3, l94ll, the patientcan)eunder our care.At that time, alnlost a nonth aftcr surgicalintervcntion,a nrassthe sizeclf a tangerinewas
found in the right side of the abdomen.Basedupon her analyses,the patient was trcated with ',a cc, glyccrol and 2 cc. of butanol solution thre.e
times a day. Shc continuedthe samc trcatmcnt without interruptionfor
501
Fto. 177. Antcroposteriorview of pelvis in case E.M. fifty-scvenmonths after onrel of treatment,
showingalmostcompleterestitution
of bone structrlre.
F'uttt',4 cids
C'otrjugatetl
ln 1947, rve startedtherapcutictrials of conjugatcdfatry acids,lirst
using eleostearicacid, the conjugatcdtrienc obtainedfrom tung oil, ad'fhc
ministeredorally or parr'nterally.
elTcctsupon pain, systemicchanges
and particularlytumor evolution,werenot up to cxpectationalthoughsubjectivcchangeswcre immediatelymorc nranifestthan for unconjugatcd
fattv
acids. [n a short timc, howevcr.it was found neccssaryto continuously
increasethe dosagein order to niaintainthe effects.'I'he intcrventionof a
defensemechanismagainstthese prcparationsoftcn rvas evident. Canccr
patients who had responcledto administrationo[ ulcostearicacid with
relicf of pain and even with an arrest of tuntor grou,th rvcre founcl to
requireincrcasingamountsof this substance.Aftcr a rvhilc.they no longcr
501
RtiSl:AR('l{
lN
Pl{YSl()PAl
ltOl.o(i}
T H T R A P N [ J T I CA F P R O A C H T O C A N C E R
503
F r c . 1 7 8 . l - a t e r a l v i e w o f t h e s k u t l o f p a t i e n t ( t l . . f , ) u t t h e t i n r c o f a d n r i s s i o nr, h o w i n g m u l t i p l e o r t e o l y t i cm e t a s t & $ e s .
504
R r . s l - t A R C l t{ N
pHvstopAlnot.oc't'
T H E R A p E L T T T cA F p R O A C H T O C A N C E R
505
r n o n t h so f l r c a t r n e n l .
506
RTsEARCH rN pHvsrclpArHot.0fir,
l : : t t ; . l f { I . , . \ r t t c r o p c r rrlieo r v i c r v o f t h e p el l i r a n t l f e n r r r r st r f p a l i e n t { l l . - f . ) t f l e r f o u r
n r o n l h r o f t r e u l n r c n t .r h e i r v i n qt h e l e l i c r n sd e c r c l r i n g ,
T H L R A P L L ' T I C
A [ tP R O A C H
T O
C A N C E R
507
two wceks shc was out of bed and resumccla nornral life. She continusd
with the same treatmcnton an ambulatorybasis.An X-ray cxanination
four monthslater showeda manifesthealingof thc previous lesions.ff i.rt.
I 84)
After anothcr four nreinthsholcvcr. without ;lnv recurrcnceof her
malignancythe diabctesinsipiduscriuld not bc adcquutcll'conlrollcd.Shc
r e f u s e dh o s p i t a l i z a t i oann d l e i t o u r c a r c .S h e d i c c lu s i r o r tt i n r c l a t c r u ' i t l r
symptomsof clcctrolyticoffbalancc.
M r . L . N . , 6 . 1y e a r so l d , h a d r l u n g h i s t o r vo f v c s i c a lt r o u b l e s w
. ith
508 I
rrEsEARcH
tN prrysropATlror.ocv
. ' 1g r ' t t l r
,'$
t
#;:
'*,
T H E R A p E U T T CA p p R O A C HT o c A N C E R
509
F r c . 1 8 4 . l - a t e r a l v i e w o f t h e s k r i l l o f p a t i c n t { S . T . ) s h o w i n g m o s l o f t b c l e r i o n sd i s appeared.
5t0
R L S E A R C H I N
P H Y S I O P A T H O L O G Y
l 1 1 , .I h i
i . t . l r i . t r 1 ; r I , ; ' l r c r i ! r r t i l t t L n , i r l g n r r c ; t f u i n n f t i ltr' l
t h e b l a d d e r .s h o w i n g t h e d e s t r u c t i r r no f r h e l c f t i s c h i o n h o n e .
T H F . R A P I L I T I C
A P P R ( ) A C I I T ( )
C A N C E R
5il
Parallel to thcse eflorts to lind new agentsfor patients with the one
typc of offbalance,othcr agentsfor usc againstthe opposite type of ofTbalancewerc investigated.
Sterolsand nonsaponifiable
fractionswere treatcd in variousways to
obtain heterogeneous
substanccs
not founelin living organisms,Wc used
heat at 300'C or ultravioletlighr, accordingro thc prcrcedures
employcd
by Roffcl.in ordcr to inducc changcsin sterolswhich would tcrnd.accord-
Flc. 186. Anteroptlttericlr view of the pelris of patient of Fig. lll.\. d rneinthr luter.
showing the healing of thc lesion.
512
x E S E A R C Hr N p H y s t o p A T H o L o c y
T H E R A p E U T T cA p p R o ^ c H T o c A N c E R
513
tensionas the criterion, and lately we have usedthe changesin serum and
total blood pntassium.In general,we administeredthe medicationonly if
the sulfhydryl index was below 1.5, the surface tension above 68. A
marked influenceupon the tumor itself was seen in a relatively high proportion of cases.The resultsalso are of theoreticalinterestsince the compound had lessinfluenceupon symptomsat the tissuelevel,such as pain,
or at the systemiclevel,and more on thoseat the cellularlevel.The effect
on pain was slow to appear,often requiringdays.But once relicf of pain
was achieved,it persistedfor a long time, in contrastto the brief effect
p r o d u c e db y o t h e r a g e n t ss u c h a s t h i o s u l f a t eo r h y d r o p e r s u l f i d casc t i n g
d i r e c t l y a t t h e t i s s u el e v e l . P r i m a r i l yb e c a u s eh e x y l d i s e l e n i dacl t e r s t h e
patternpresentat the cellularlevel,it must bc emphasized
that determining
proper dosageat least in the beginning,appearedmorc difficult than for
any other substance
with which we have had experience.
A t t h e b e g i n n i n gw
, e u s e dd o s e si n t h c r a n g eo f l 0 - t t 0 m g . b u t i n a
numberof casesa pcrsistentchangeto the oppositcpatternoccurredafter
o n e o r t w o d o s e sT
. h i s l e d t o t h e u t i l i z a t i o no f s m a l l e ra n d s m a l l e rd o s e s
in order to avoid too rapid changcto the oppositepattern.Also with small
d o s e sw
, e h o p e dt o l i m i t t h e t h c r a p e u t ircc s p o n s teo t h e c e l l u l a rl e v e lo n l y .
w e d e c r e a s ctdh e d a i l y d o s c t o 5 m g . , t h e n t o I m g . a n d e v e n t u a l l ye v e n
t o m i c r o g r a m sW
. i t h t h e s es m a l l a m o u n t s .t h e i m m e d i a t ec l i n i c a le f l e c t s
s e e m e dt o b e a l m o s te n t i r e l yl i m i t e dt o t h e c c l l u l a rl e v c l .T h e c h a n g e si n
pain and the systenric
patternwereminimal.The choiccof doseto be given
w a sd e t e r m i n c idn t h e s e c o n dp a r t o f t h i s r e s e a r c p
h r i m a r i l yb y u r i n a r ys u r face tcnsion.It was observedthat microgramdosesmay influencelesions
without changingthe valuesof the other analysescorrespondingto the
systemiclevel.
The clinicalresultsobtainedwith the useof hexyldiselenide
in humans
warrantsdetailedconsideration.
Cood resultswere obtainedwith greater
consistencyafter the problem of dosagewas resolvedand the relationship
to urinary analyseswas established.
Important objectivechangescould be
achievedby using urinary analysesas a guidefor dosage.as illustratedby
the following cases.
A . 8 . , 6 8 y e a r so l d , m a l e .I n O c t o b e r1 9 4 9 .t h e p a t i e n th a d a n a m p u tation of the left leg at the hip-joint performedat the Memorial Hospital
becauseof a tumor of the femur. The pathologicaldiagnosiswas sarcoma.
H e w a s w e l l u n t i l J u l y 1 9 5 5 w h e n h e b e g a nt o c o u g ha n d h a v e r e c u r r e n t
e p i s o d eo
s f h e m o p t y s i sX. - r a y e x a n t i n a t i o rne v e a l e dr i g h t p l c u r a le f f u s i o n
a n d i n f i l t r a t i o no f t h e r i g h t l o w c r l o b e . H e w a s r e u d n r i t t e d
to Memorial
Hospital in october 1955 and thoracentesis
revealedbloody fluid. Bron-
514
./
n r , s E A R C Ht N p n y s r o p A r H o l , o c y
chr:scopy
showcdpartialnarrowingof the right lower lobe bronchus.Examinatiein
of pleuralfluid and bronchialwashingby the Papanicolau
methodshowedceltsthat wcre suspicious
for mirlignantdisease.but not
canclusivc.
It couldnertbe dctermincd
rvhether
a primarylung canceror
metastatic
silrconta
wasprescnt.An exploratory
thoracotonly
wasadviscd
but refuscdby the patient.who signcdhimsclfout.
2-5-s5
f r t t ; . l l { 7 . r \ n l e r o p o s t c r i r t rr ' r c u r r f c h c r t r n ( ' ; r s c { . 4 . 8 . ) a t t i n r c o f a t l n r i r s i o n s h o * . i n s
l h c p r e r c n u : co f l r m a s r i n t h e k r w e r p a r t o f t h e r i g h t h c n r i r h o r u x .
THf-RAPI:-t;TI(
4t'PRt)4(lll
t'() ('AN(:hR
5r5
,.*.."
*Wrt
.,,.rsqffiF*
".*n
,r'
a - 2 6- 5 6
iYR*.
,$
rs:
';*
F t c . 1 8 8 . A n t e r o p o \ t e r i o rv i e w o f t h c c h u r t o f t h e p a t i e n t . { . 8 . a f t e r t r . n m o n t h s
o f t r e a t m e n tw i t h h c x v h l i s c l e n i d c .
At the beginningof Februarv.thc nrassin the right lrls'cr lobe was alnrost
entirelygonc. The intcrlobularthickcningof thc left ktucr lung still pers i s t e dl.n A p r i l . f u r t h e rc l e a r i n go l t h c r i g h tl c l w c rl o b c w a ss s c n .l f i S . 1 8 8 /
for a fel morc nronths.
The patientreceivcdtreatnlcntwith hcxyldisclenidc
H e c o n t i n u e di n e x c e l l e nct o n d i t i o n* i t h o u t f u r t h c rt r e a t m c n ft o r t h e n c " t
three years.At thc cnd of that tinrc. thcrc tus il recurrcnceof pain and
thc massin thc right lowcr lobe rcappearcdlnd grcw rlpidly. Trsatment
was rcsumcd.Thc pain disuppearcdagain within a
with hcxyldiselenide
shorttime. and thc tumor againregrcsscd.
516
x E S E , A R c Ht N p H y s l o p A T H o L o c y
T H E R A p E U T t CA p p R O ^ C H T o
C A N C E R
5 1 7
tcnsioninto the renal vesselsand surroundingtissue."There was no evidenceof pulmonaryor other metastases.
The patient did not undergo any treatment at this time and felt well
until September1955, when he experiencedincreasedfatigue,flatulence,
slight pain in the right kidney region,and some abdominalpressure.No
abdominal mass or lymph node involvementwas found. He was treated
with hexyldiselenide,
starting with 200 microgramsa day. This dose was
increasedprogressivelyuntil it reachedZVz mrlligramsa day.
Under this treatment, most of the patient's pain disappearedand no
palpablemasscould be found. The patientcontinuedtreatment,with the
samedose,until October 1957,when epichlorohydrinwas added.At first,
with the new rnedication,the patient was more tired and there was a noticeablcincreasein perspiration.However,after a short time, hc continued
to improve.At present,he is still taking l5 milligramsof epichlorohydrin,
and 100 microgramsof hexyldiselenide
daily. He feelswell, continuesnorwork,
palpable
mal
and no tumor is
now four and a half years since the
beginningof the treatment.
W. H.-ln March 1954,at the ageof I l, this boy had the first of three
brain operationsat the JerseyCity MedicalCenter.He had complainedof
persistentheadaches
for about a year.A brain tumor was removedand on
pathologicalexamination,was first thoughtto be benign,but later proved
t o b e m a l i g n a n t( s p o n g i o - b l a s t o m aB) .y A u g u s to f 1 9 5 4 , h i s s y m p t o m s
recurredand a swellingappearedin the area of the scar. At the second
operation,it was possibleto removeonly a part of the rccurrenttumor. In
November 1954, local swellingand headaches
returned.X-ray treatments
failed to give relief and a third operationwas performed,but only a piece
of tumor was removed.lt showedthe sanrepathology.
H e c a m eu n d c ro u r c a r eM a y 3 , 1 9 5 5 ,a n d w a st r e a t e db, e c a u s e
of low
with
hexyldiselenide,
sulfhydrylurinary indcx,
in dosagesrangingfrom 300
-l'his
microgramsto I milligrampcr day.
treatmentwas continuedfor one
year. He has done rcmarkablywell since.He has had no headaches,is
more alert and lessdrowsythan at any time sincehe first becameill, There
has been no evidenceof recurrence,Since treatnrcnt,there has been a
definitereductionin involuntarymovementsof his head, extremitiesand
body, which appearto havebeen relatedto the tumor growth. There have
to date,five yearssincethe beginningof
beenno abnormalmanifestations
the treatment.The patient goes to school and engagesin all the activities
of a normal boy of his age,
Mrs. A. L.-This patientfirst noteda lump in her neck in the summer
of 1953 when she was 26 years old. In February 1954, a secondlump
518
R E s E A R c Hr N p H y s r o p ^ T H o L o c y
appeared.Surgerywas performedat the Ottawa Civic Hospital.The pathologicalexaminationshoweda cancerof the thyroid (papillary adenocarcinoma). It involvedboth sidesof the glandand many of the lymph nodes.
The entire thyroid gJandwas removedand bilaterallymph node dissection
was done in two stages,Sincethe surgeonfelt that he had not rcmoved
all the affectedareas,the patient receiveddeep neck X-ray therapy following the operation.However,by July 1954, a new massdevelopedon the
right side of the neck and this was removed in November.Thc pathologicalexaminationproving it to be the same type of tumor.
T h e p a t i e n tw a s f i r s t s e e nb y u s o n J a n u a r y1 0 , 1 9 5 5 .A l t h o u g hi t w a s
only two months since the last operation,there were severalrecurrent
tumor massesin the lateralright side of the back of the neck, as well as
infiltrationsinto the area of the last operativewound. The patient was
treatedwith hexyldiselenide,
in dosesrangingfrom 300 microgramsto I
milligramdaily. Under this treatment,the massesprogressively
decreased
and, after 3 months,disappeared.
Shecontinuedthe treatmcntfor another
six months.Now, after 5 t/z years,she is feelingwell. has had no recurrcnces,and is carryingon her usual activitiesas a housewifeand mother.
Hexyldiselenide,
althoughit producesimpressiveresults,very often is
not of itself able to provide enduringbenefit.Many other patients have
had recurrences,
some in spiteof impressivefirst resultsand continuation
o f t r e a t m e na
t s s h o w ni n t h e f o l l o w i n go b s e r v a t i o nJ.. D . , l 0 y e a r s o l d ,
cameundcr our care in a preterminalstate,aftcr an exploratorylaparotomy
revealingan extensivecarcinomaof the liver. (Fig. 189) The responseto
hexyldeselenide
treatmentwas impressive
with the patientmaking a perfect
rccovery.His livcr which had filled the abdomen,returnedto normal dim e n s i o n sT. h e p a t i c n tc o n t i n u e da n o r m a ll i f c f o r 2 y e a n sw h c n , i n s p i t e
of the continuationof the treatment,gcneralizedrecurrencesappcared.
Thesccould no longerbe controlled.
Tetralin Perselenide
The good cffectsobtained with persulfideson one hand, and with
seleniumon the other, have led us to investigate
the correspondingcomp o u n d - t e t r a l i n ep e r s e l c n i d cw.i t h i t s l o w t o x i c i t y ,t h c c o m p o u n dw a s a d m i n i s t e r c dt o h u m a n si n w h i c h t h e d e s t r u c t i o o
n f t h e t u m o r a p p e a r e dt h c
immediateaim. The cffectupon pain was good althoughnot immediate,thc
s a m ea s f o r t h e s y s t e m i cl e v e l .I t w a s a t t h e c e l l u l a rl e v e l sw h e r e t h e s e
were the most manifcst.Researchwith this agent is still in progressand
for the momentthe influenceexertedupon the tumorsseemsto be very fa-
.IH[:RAPEU'TIC
APPR0^CII TO
CANCER
519
F r c . 1 8 9 . P h o t o m i c r o g r a p ho f a l i v er h i o p s yo f p a t i c n tJ . D .
520
xESEARcHrN pHysropATHoLocy
l.llllRAPF.trTl(
At,t'R(]A( l{
1'o
( ANctiR
5lr
F t c ; . 1 9 0 . P h u t o n r i c r o l i r a p ho f i r l r n t p h n o d e o f p u t i c n t \ 1 . S . s h o ! , , ' i n ga l - v r n p h o .
\a rconta.
not cntirely control thc condition. It was rvith highcr alcoholsthat irn-fhc
portant objectivcas well as subjectivcchangeswcrc achievcd.
tunlor
decreased
rapidly. Intcrruptionof treatment.duc to fie cardiaccondition"
did not seemto inlluencethe favorablcproercssof the condition.
M. B.-Toward the'end of 1950.at thc agc of 4-5,thc patientbcgan
to complain of right lower quadrant and i.ntcrnrittentlcft upper quadrant
p a i n .S h cd i s c o v e r e a
d n a b d o n r i n anl r a s sb y h e r s c l f I. n F c b r u a r yl 9 - 5 1 .a n
exploratorylaporatomywas pcrformedat Lsnox Hill Hospital rvith the
following findings: "Situated in the left side of the pelvis, a huge cystic
structurcthe sizeof a football was found. 1'hc supcrioru'all oi this nrirss
was attachedto thc nrcsentery
of thc kxrp of the snrallbori'cl.The anterior
522
nEsEARcH tN pHystopArHoLocy
.i
-523
F r c . 1 9 l . P h o t o m i c r o g r a p ho f a l y m p h n o d e b i o p s i e df r o m p a t i e n t M . B . s h o w i n g
meta$taticadenocarcinoma.
524
nEsEARcH rN pHystopATHoLoGy
and for four years has had no subjectivediscomfort.No mass can be felt
in the aMomen and the patient is well now.
Radiation and Chcmotherapy
Parallel to the guided chemotherapy,the idea of mixed radio- and
chemotherapyhas led us over the yearsto severaltentatives.In one of them,
in collaborationwith Dr. L. Goldman on a larger scale,chemotherapywas
addedto radiotherapy.(Note I)ln spite of the interestingimmediateresults, l/ chemotherapywas not further pursued, the ultimate fate of the
patientsthus treatedwas no better than that of the patientswho had received radiationalone. Analysisof thesevarious attemptshas permitted
us, however,to establishthe conditionsunder which such a mixed treatment would appear especiallyindicated.consequently,it is the type of
offbalancepresentwhich is seento determinethe relationshipbetweenthe
two therapies,and respectivelythe procedureto be followed. The samc
relationhas permittedto definethe conditionsunder which chemotherapy
would be indicatedand addedto radiotherapy.It should not be forgotten
that throughits action upon the fatty acids,radiationrepresentsindirectly
an agentof the samegroup to be addedas the negativelipoids.
The caseswith a type A offbalancewhich seemto respondinsufficiently
to medicationwith agents,representan indicationfor the mixed treatment.
Acting in conjunctionwith radiation,chemotherapy
has appearedespecially
active. Chemotherapystartedbefore,is continuedactively during radiotherapyand followingradiation,for a long time, even for years.The radiation acting as an adjuvantagentis usuallyapplied in small doses.fronr
only severalhundredr to a few thousandr, in generalmuch belorvthosc
dosagesknown to induceby themselves
alone therapeuticeffect upon tumors. In caseswith type D, as the radiotherapyrepresentsan agent increasingthe actualoffbalance,
an addedchemotherapy
with positivelipoids
would have the role of correctingnot only the existingmanifestations
but
of preventingsome of those which would be inducedby radiotherapy.It
w o u l d i n f a c t h a v e t h e s p e c i f i ca i m t o p e r m i t t h e c o n t i n u a t i o no f r a d i o t h e r a p yw h i l e l i m i t i n gi t s n o x i o u si n f l u e n c e .
The mixed therapyhas to be guidedby the routinetest, with frequent
analyses,in view of the rapid important changeswhich have been seen to
occur. In thesecases,it appearsusefulto follow especiallythe changesof
the ctrlorideindex, thoseof peroxidesin the urine which togetherwith the
surfacetension,are seento be not only particularlyinfluencedby radiation
but able to indicatethe importantchangein offbalance.
THERApEUTIC AppRoACH
'ro
CANCER
525
526
RESEARCHtN
pHysropATHoLocy
T H E R A p E U T t cA p p R o A c H r o
cANcER
527
the cellular level, and bixine and propionic aldehydeat the organic and
systemiclevel-the conditionseemscontrolled.I'he pulmonarymetastases
seenin previousX-rays have alsodisappeared
and all thc skin nodulesdecreasedrapidly, most of them having complctelydisappeared.she is at
presentin good stateof health,doing all her own housework.
Mrs. c. H., 60 yearsold, camc under our care two years ago with a
historyof lymphosarcoma
with two positivebiopsiesof thc inguinalglands,
complaining
e s p e c i a l lo
y f p a i n i n t h e a b d o m c nO
. n e x a m i n a t i o ni ,n g u i n a l ,
axillar and cervicalglandswere present,someof thenr 4 crn in diameter.
With treatmentof sodium thiosulfateand fatty acid hydropersulfides
in
relativelysmalldoses,the pain wassufficiently
controlled.After two months
of treatment,the patientexperienced
extremepain and a tumor of the head
of the right humerus.In view of the local lesionswith analyseswhich all
showedan intensiveoffbalanceof type A, the treatmentwas changcdto
epichlorohydrinand heptyldiselcnide.
Administeringtheseagentsin relativelyhigh doses,it was possiblcto not only controlthc pain in a few hours,
but to have the iesiondisappearin lessthan two weeks,as did also all the
abnormalgJands.
However,in spiteof theseobjectivevery good results,the
generalconditionbecameunsatisfactory,
cspeciallywith marked weakncss.
C h a n g e si n t h e d o s e so f m e d i c a t i o o
n r t h e t e m p o r a r yd i s c o n t i n u a t i oonf t h e
m e d i c a t i o nf.a i l e dt o c o r r e c ti t . I t w a s o n l y w h e n p r o p i o n i ca l d e h y d ec o n s i d e r e dt o a c t u p o nt h e o r g a n i s ml c v c lw a sa d m i n i s t c r e tdh. a t a r a p i dc h a n g c
t o w a r da f e e l i n go f g e n e r aw
l ell-being
w a s o b t a i n e dA. t p r e s e n tw
, ith small
dosesof propionicaldehydethe patientis entirclyfree from any subjective
or objectiveabnormalmanifcstations
and has bccn so for almost a year,
M r s . E . R . , 4 2 y e a r so l d , h a d a r a d i c a lm a s t c c t o m fyo r a n a d e n o c a r cinomaof the breast.After I t/z years,shc experienccd
persistent
back pain
w h i c hw a sf i r s td i a g n o s e ad s a r t h r i t i sX
. - r a y s t u d i c sa n d a m y e l o g r a mm a d e
at MontefioreHospital showedtwo metastaticlesionsat the I st and 2nd
lumbar.Surgicalinterventionwas thoughtto be not indicated.X-ray therapy
neitherrelievedthe pain nor made it possiblcfor her to lcave her bed. At
a d m i s s i o un n d e ro u r c a r e ,t h e p a t i e n w
t a si n s e v e r ep a i n a n d u n a b l et o e v e n
turn in bed, althoughshewas still ablc to moveher legsand thcir sensitivity
w a s c o n s e r v e dA. t r e a t m e nw
t i t h h e p t y l d i s c l c n i dpcr,o p i o n i ca l d e h y d ea n d
bixine made thc pain disappearand up to date the patient is leading a
normal life, after havingbeenbedriddenfor 8 months.
M r s . M . M c B . , 6 2 y e a r so l d , c a m c u n d e ro u r c a r ef o r a b a s o c e l l u l a r
carcinomaof the left side of the face,near the inferior eyelid.Advised to
.,..
,.:,:,.4t
:ltt ,
'.:j:.,!:;:ti
528
RESEARCH tN
PHt'StOPATItOLoCY
undergosurgcry,which would cnucleatcthe left cyc. shc rcfusedthe operation. A hiopsy u,as performcd (l'ig. tq2) showing the presenceof a
basocellularcarcinoma.She u,as treatr-'dlvith hcxyldisclcnidcand with
sodium thiosulfate,and the lesion clislppearedwithin a few weeks. No
rccurrcnccrr,asobscrvcddurint thc nlrsl 6 r'cars.
l"l
. t
r,s
t;r'*
tl"
rl
ti
rd
'r\
A
F l t ; . l 9 l , P h o t o m i c r o g r a p ho f t h c h i o p : y n r l d c o n p a t i c n t l t l . N l c B . s h o w i n g u
b l s t l c e l I u l a rc u r c i n o l n t r .
M r . C ' 2 . . 5 6 Y c a r so l d c ; . t n t u
c n d c ro u r c a r c f r r r a l c s i o no f t h e u p p e r
l i p . B i o p s y p r o v c d i t t o b c x s q u a n l ( ) ucsc l l c a r c i n o m a . ( f i g . / 9 J l T h e
patientrefusedsurgeryor nrdiation*hich r,''cadvised.Followingthe biops.v
t h c t u n r o rs t l r t c d t o s r o $ r ; r p i d l r .u \ s c c ni n F i g . 1 9 4 . A c c o r d i n gt o t h e
anall'scs.
the paticnt*'us trertctl rrith flttv acid hydropersulfidc.
The lcsion
(
i
n
d i s a p p c a r c d l c s st h u n I u c c k s t : t g . 1 9 ,/5N o r c c u r r c n c cw a s o b s c r r v c c l
a l t h o u g ht h e p a t i c n tw a s n o l o n s c r u n d e r l r c a t m e n t T
. h c p a t i e n td i e d i i
m o n t h sl l t c r f r r l n t i t c ( ) r ( ) n i l r ov c c l L r s i o n
"l'hcability to induccchangcsat sinqle
specificlevelshas alteredcompletclyi,l'hatrvus()nceit grossduulistrcapprolch. Llntil rccently,we had
c o n s i c l e r ci d
t i n l c l r i s a b l el o i n d u c et h c r l p c L r t icch a n g c su t a s i n g l el c v c l i f
r-rflbalances
\\,crcprcscntat nrorethan onc levcl. With the conceptof level
independcncc.it has become part of the merhod to attempt to influcncc
T T I E R A P L U T I C A P P R ( ) A C I I" T OC A t \ i C I R
l9l.
P a t i e n t ( i . 2 . h e f < t r cl r c a r r n c n t
F t c ; . 1 9 4 . T h c t u m o r g r c w r a p i d l l f o l l r r u i n gh i o p s y
F r ; . 1 9 5 . P a t i e n tt i . Z , a f t er r r c ; r r n r c n r
-5l()
530
* E S E A R C Hr N p H y s r o p A T H o L o c y
C H A P T EI R6
PRESENT F-0RM
OF TREATT{ENT
rTr\t
l L n , , " u * A p E U r r c M E A S U R EaSg a r n sct a n c c rw h i c h h a v e e v o l v e dt h u s
far out of the pathogenicand pharmacodynamic
conceptsdescribedin these
pageshave provcd, as we have shown above,to bc useful.All along in
their development,thcy have fallen short of producinguniformly,the full
measureof benefit which, we have continuouslyhoped, one day will be
attainedin most and possiblyin all casesof malignancy.
Yet the resultsachicvedwith biologicallyguidedtherapy,imperfectas
t h e y h a v e b e e n , h a v e c o n t i n u o u s l yi n d i c a t e dt h c g r e a t p o t e n t i a lo f t h e
method.
W h a t h a s b e e na c c o m p l i s h eddu r i n gt h c y c a r sh a s b c e n h a r d l yc o n s i d ered as a linal measureof the effectiveness
of this basicnew approachto
treatmentbut onJy of the availablecriteria and agents.With thc criteria
and agentsthe method itself has been evolving.In its presentform, the
method representsa valuabletool for helping many cancer patientswho
are entirelybeyondhelp with presentday methods.In spite of its accomplishmentsfurther advancesalong the same lines-both in techniquesfor
recognizing
fundamentaloffbalances
at diflerentlevelsof organizationand
cven in new compoundsmore effectivein correctingthem-are to be
expected.
Equal in importancewith the current resultswith biologicallyguided
therapyis the fact that new applicationsarc evolving;that this approach
helpsus to betterunderstanding
and treatmentof malignancyand of other
pathological
well;
conditionsas
and that, by its very nature,the methodhas
the capacityto furnish the guidanceneededfor improvingit.
We are presentinghere the form of treatmentwhich we currentlyuse.
531
: 1 , :
.
| . .
532
n E S E A R C Hr N P H Y S r o p A T H o L o c y
OffbalanceD
high
low
low
high
low
low
low
low
high
Iow
high
alkaline
Average
Values
I .016
6.2
68
4.5 mEq
38 mEq
37'C
7000'cmm
1 0 0/ c m m
, , I
2.5
s::'s
Offbalance
low
high
high
low
hish
high
high
high
low
high
low
acid
PRESENT FoRM
oF
TRE^TMENT
533
534
n E S E A R C Hr N P H y s r o p A T H o L o c y
PRESENT FORM
OF TREATMENT
,,i .535
Caffeine
Procaine
('odeine
Morphine
Dcnrcrol
A m i n o p h yl l i n e
Liver extracts
vit. A, D, 8,,
I ron
Vit. 8,, B,, K. E
Testosterone
I)rogesteronc
Sti Ibesterol
Dcsox.v-cort
icoslerone
G I ucosanri ne
Epinephrine
Penicillin
streptomycin
Aureomycin
Cortisonc
Sulfas
Insulin
B a r bi tu r a t e s
lv{ercuhydrine
Benzedrine
B e na dr r I
536
pHysropATHoLocy
xESEARcH tN
In tuing the various agents,we have to keep irt mind the eftect upon
the existingoffbalances.
Conductol Treatmenl;Although there may be somevariationsdepending upon the circumstancesof individual cases,treatment is conducrcd
generallyas follows:
Urine and blood analysesare performed.Values for two or three consecutivedays are usuallydeterminedbeforestartingtreatment.If all values
indicate the same pattern, the diagnosisof the offbalanceis clear. If the
analysesindicate the presenceof different patterns, an interpretation is
made on the basisof analysesshowingthe offbalanceat specificlevels.For
patients in whom manifestationsinvolve the metazoic compartment, urinary surfacetensionis the important criterion. Urinary specificgravity and
urinary pH, are important when systemicor tissular manifestationsare
present.Potassiumis the criterion when changesat the cellular level are
most important.The body temperatureand the other complementaryanalysesare usedwhen discordantpatternsare encountered.
They help to recognizethe type of offbalanceat the different correspondinglevels.
Once the level oftbalancesare determined.suitabletreatmentis instituted.The agentis chosenfrom the proper group accordingto the level
TrsLE XXIV
Acexrs CHosrNAcconorNcro rHE Tr,srs
AceNrs
Test
Level
OftbalanceA
OffbalanceD
Cellular
Potassiumin I S e l e n i u mp r e p a r a t i o n s Heptanol
blood
lEpichlorohydrin
Urinary
calcium
Tissue
U r i n a r yp H
Lipoacids
Blood
tTetralinpersulfides
eosinophiles
Hydropersu
lfides
Mg Thiosulfate
Surface
Tension
P a i np a t t e r n
Polyunsaturated alcohols
Unsaponifiable
fraction
liver
Glycerol
Butanol
Organ and
U r i n e s p e c i f i cM g o r N a T h i o s u l f a t e
gravrty
Organisnr
P r o p i o n i ca l d c h y d e
Glycerophosphoric
or
I
I
Surface.
l ensron
I Body
l_
I
t"lo"'"i"':]
lactic acid
PRESENT FORM
OF TRE^TMENT
537
538
*ESEARcHrN pHysropATHoLocy
P R E S E N T F O R M O F T R I ] A T M E N T
539
540
REsEARcH tN pHysropATHoLocy
ness.For 30 years,the patienthad complainedof pain in the right hypochonder,related to the prescnceof gall btadder stones.In February of
1960, a laparatomywas performedand a tumor of the gall bladder with
metastases
to liver was found. Only a biopsywas performedwhich showed
a carcinoma.At admissionunder our carc, the patient was in very poor
generalcondition with marked dyspnea,dcep jaundice and severepains
in the upper abdomen,emaciatcd.A right pleural effusionwas found and
a thoracentesis
performed.An irregular masswas found in the right hypochondriumarrivinguntil the umbilicus.Shehad clay coloredstools,typical
for obstructivcjaundice.In spiteof thoracentesis
the dyspneacontinuedto
be severeand the patientwas kept under oxygen.The patient was placed
under chemotherapeutictreatment with epichlorohydrin,bixine and perselenidein accordance
with her urinalyses.
The patient'sconditionimproved
progressively.
The stool returncdto normal color; fluid in the right chest
did not reproduceand the mass on the right hypochondriumdecreased
p r o g r e s s i v etloy h a v et h e l i v c r i n n o r n r a d
l irncnsionA
s .t p r e s c n tt,h e p a t i e n t
with all the subjectiveand objectivcsympromsimprovcd considerably,is
ambulatory.
M r . S . S . , 6 4 y e a r so l d , w a so p e r a t e di n l 9 _ 5 3f o r a h y p e r n e p h r o moaf
the right kidney.Two yearslater, massivemetastases
were seenin the left
femur and pelvic bones.Pathological
fractureof the neck of the left femur
was treated surgically.Further X-ray cxaminationsrevealed extensive
metastases
of the femura,and pelvicbones,with multiplelung metastascs.
When the paticnt came undcr our carc, he was sufferingagonizingpain
in the left hip. An X-ray cxaminationshowedan almostcomplete
especially
disappearance
of thc upper part r)f thc lcft fenrur,with multiple metastases
in the right femur and pelvicbones,and multiplenretastases
in both lungs.
Accordingto the analyses,
the patientwas treatedwith perselenide,
bixine,
and epichlorohydrin.The condition improvcd rapidly, the patient being
ableto sit up and evento walk a little. Hc rvasdischarged
from the hospital
to follow the treatmcntat home which was done very irregularly.He was
readmittea
d n r o n t hl a t c r w i t h v e r y s e v e r ep a i n a n d t h e t r e a t m e nrt e s u m e d .
g r a d u a l l ya n d t h e g e n c r acl o n d i t i o ni m p r o v e dm a r k e d l y .
The painsubsided
RccentX-rays revealeda manifestrecalcification
of the upper part of the
s ,a d n o l o n g e rb e e nv i s i l e f t f e m u rw h i c h ,i n p r e v i o u sX - r a y e x a m i n a t i o n h
ble. At the sametimc, many of the metastatic
lesionsof the lung disappeared
while in others,a markeddecreasein their sizewas seen.Thesesubjective
and objectiveimprovementsare continuingconstantly,up to date.
F . G . , 6 1 y e a r o l d f e m a l c .i n J u l y 1 9 6 0 h a d p a r t i a lc e c o s t o m yf o r
The mcsentericlymph nodes were found involved. A
adenocarcinoma.
P R E S E N T F ' o R M o l ' T R E A T M E N T
5 4 1
NOTES
C h a p t e rI , N o t e l . S u b n u c l e a O
r rganization
T h e a n a l y s i os f t h e a v a i l a b l e
d a t ac o n c e r n i nsgu b n u c l c apra r t i c l c ss h o w s
that while an unsuspccted
number of diffcrentparticlesari progressively
discovered,no satisfactoryrelationshipbetweenthem-from the point of
view of the organization---can
be established.
The rccognitionof a pattern
c o n c e r n i n gt h e i r o r g a n i z a t i o n ar le l a t i o n s h i p
w o u l d f i l l a n i m p o r t a n tg a p
in the knowledgeof this cntire field.Thc fact that the samepatterngoverns
t h e o r g a n i z a t i o ni n g e n e r a l ,f r o m a t o n r i cn u c l c i u p , h a s i n d u c c d u s t o
a t t e m p tt h r o u g ha n c x t r a p o l a t i o nt,o s c a r c hi t f o r t h e s u b n u c l e a re a l m .
A s s e e na b o v e .t h e s t u d yo f t h c o r g a n i z a t i ohna s p c r m i t t c du s t o d c f i n c
t h e f o l l o w i n gc o n c e p t sa s c h a r a c t e r i s tfi o
cr the organizationa
p la t t e r n :
I ) A l l t h c e n t i t i e si n n a r u r ec a n b e i d e n t i f i e db y t h e i r p l a c e i n a
h i e r a r c h i co r g a n i z a t i o ni,n w h i c h t h c c n t i t i c sa r c c o n n e c t c dt h r o u g h a
s u p e r i o r - i n f e r i or re l a t i o n s h i pA. n e n t i t y e n t e r si n t h e f o r m a t i o no f o t h e r
e n t i t i e sw h i c h a r e c o n s i d e r e d" s u p e r i o r "t o i t . a n d i s f o r n r e db y e n t i t i e s
w h i c h a r e h i e r a r c h i c a l l"yi n f e r i o r " t o i t .
2 ) E a c h e n t i t y i s f o r m e d b y a p r i n c i p a la n d a s e c o n d a r yp a r t , t h c
p r i n c i p a lp a r t b e i n g r e p r e s e n t ebdy e n t i t i e sh r e r a r c h i c a l liyn f e r i o r t o i r ,
r v h i l et h e s c c o n d a r yp a r t , b y c l c n r c n t st a k e nf r o n r t h c i m m e d i a t cc n v i r o n m e n t i n w h i c h t h e e n t i t i e sf o r m i n gt h e p r i n c i p a p
l a r t h a v ee x i s t e d .
3 ) E n t i t i c sw i t h s i m i l a rp r i n c i p a lp a r t sb c l o n gt o a s a m cl c v c l . I n t h c
h i e r a r c h ipc r o g r c s s i o nt h, e r ea r ee n t i t i e so f t h es a m el e v e lw h i c ha r eg r o u p e d
t o g e t h etro c o n s t i t u t e
t h e p r i n c i p a lp a r t o f a n e n t i t yo f a l e v e li m m e d i a t e l y
superior.
4) From the energeticpoint of vicw, thc principalpart in the organization of eachentity appearsmore positivethan the secondarypart to which
it is bound.
5 ) The hierarchicprogressionof the organizationfrom one entity to
t h a t i m m e d i a t e l ys u p e r i o rt o i t , i s m a d e t h r o u g ht w o p r c r c c s s ewsi t h t w o
d i f f e r e n ti n t e r v e n i n gf o r c e s .F o r c c so f c o l u m b i a nn a t u r e b r i n g a n d k e e p
the electrostatically
opposite principal and secondaryparts together,A
n e w e n t i t ya p p e a r sh o w e v e ro n l y w h e nq u a n t u mf o r c e si n t e r v e n eo, r g a n i z i n g t h e r e l a t i o n s h i op f t h e t w o c o n s t i t u e n tasn d e s p e c i a l l tyh e i r r e c i p r o c a l
m o v e m e n tT
. h e i m m e d i a t ea i m o f t h i s o r g a n i z a t i o ni s t o p r e v e n ta r e c i p r o c a lt o t a l a n n i h i l a t i o no f t h e t w o p a r t s ,p o s i t i v cp r i n c i p a la n d n e g a t i v e
secondarywhich would occur if the electrostaticforces alone would be
present.
6) In the hierarchicprogrcssivcdevclopmcntof the organizationfor
543
544
n . E s E ^ R C Hr N p H y s r o p A T H o L o c y
eachpassage
from one levelto the immediatelysuperior,thesetwo kinds of
forces---electrostatic
and quantum-were seento intervenealternatively.The
fulfrllmentof one force is seento inducethe appearanceof the other. From
an energeticpoint of view, an entity will appearinactivewhen its electrostaticforcesare fulfilled but with quantumforcespresent,or energetically
active with quantum forces fulfilled and the electrostaticforces present.
The exampleof atomsand ions is a typicalillustrationof this relaiionship
for the atom level.
As a work hypothesis,we tried to apply the above schematically
presentedconcept of hierarchicorganization,to the subnuclearrealm. lt
is only by using analogiesthat such an attemptcan be made. The scarce
data availableseem to confirm however,this view. According to it, the
electronand positronwould representthe lowestentitiesof the subnuclear
realm with the smallestmassand oppositecharge.If thesetwo corpuscles.
when attracted, encounter one another, they will annihilate each other
with liberationof two photons.This annihilationis preventedhowever,
althoughthe two corpusclesremain bound togetherthrough their electrostatic forces by the interventionof quantum forces organizingtheir reciprocal movements.The rcsult is a new entity, of a level immediately
superiorto the positron,in which the electronis kept electrostatically
bound
to the positronbut kept into an orbital movement.Hypothetically,it can
evenbe conceivedthat throughdifferences
in the resultingmovement,more
than one solutionwould exist.
Residual Clurge
Due to the interventionof this movement.the resultingelectrostatic
neutralizationbetweenpositronand electronis incomplete.A "residual"
positivechargewould characterize
the new entity.This chargealone would
not be sufficientto keep anotherelectronby neutralizingits charge.However severalsuch entitiesgrouped togethercan have the sum of their
"residua.lpositivecharge" such as to be compensated
by a new electron.
The two electrostatic
forces,that of the groupof entitiesand that of the new
electronwill keep thesetwo parts together,while the quantum forces will
again organizethe movementof this new added electron,preventingthis
t i m e a g a i nt h e a n n i h i l a t i o o
n f t h e s et w o p a r t s .A n e w l e v e l ,t h i s t i m e o f t h e
third order, is thus realized.It is casyto conceivethat severalsolutionscan
existfor each case,sincethe sum of the residualpositivechargesdoes not
Severalsolutionsappear
correspondexactlyto that of thc negativeelectrons.
thus possible.Besidesthis in which a small group of entitieswould be
by one electron,a higher number of entitieswould be kept
compensated
togetherwith the sum of their residualchargesapproachingthat of two
or more electrons.For each level,severalsuch solutionsare conceivable.
passage
toward higherlevels,the number of the soluWith the progressive
tions increases.
positiveand negativeparts of the
The fact that the two electrostatic
entitiesdo not compensateperfectly,leadsto the possibilitythat the compensationtakes place either with an excessor lack of negativecharge.
nEsEARcH rN pHystopATHoLocy
546 /
Frc.,,,r-ot-&**
Hlsttdtnc
@'-@,*
l'ilIR0GEmt6
8ASS
Prlncs
Adeninc
furlrtdincs
Thplnc
ol
I
ot
61zcrr,
,\/
H
Cytostrr
tfu-ot-o,z-o,Sro.,
ffi
H
F l c . 2 0 1 . T h e N H , a n d c - N - c - N g r o u p sa p p e a ra s e n t i t i c st a k i n g p a r t i n t h e f o r m a t i o n o f a l k a l i n e a m i n o - a c i d sa s w e l l a s o f n i t r o g e n o u sb a s e s .T h e b o n d t o a c h a i n
h a v i n g a n a m i n o a c i d g r o u p i n t h e f i r s t c a s e ,r e s u l t s i n a n e w e n t i t y - a n a l k a l i n e
amino acid-which polymerizesthrough the amino-acid group, Through the alkalinc
group it conserves its positive electrical character. In the nitrogenous bases, the
C-N-C-N group is part of the cycle. Bound to phosphoric acid, the results are acid
e n t i t i e sw i t h n e g a t i v ec h a r g e s .
NOTES
541
( o r i n f i s h ,t h e p r o t a m i n s )w o u l d t h u s r e p r e s e ntth e f l r s t h i e r a r c h i cs t e p s
i n t h e p r o g r c s s i oonf t h c b i o l o g i c asl c r i c s .
A f e w w o r d ss h o u l db e s a i da b o u tl y s i n c .t h e a l k a l i n ea m i n o - a c i dw i t h
a n a m i n og r o u p a s a l k a l i n et e r m i n a g
l r o u p .A l t h o u g ht o g c t h c rw i t h t h c
o t h e r a l k a l i n ea m i n o - a c i d si t, e n t e r si n t h e f o r m a t i o no f h i s t o n e si,t s e e m s
t o h a v e a n o t h e ri m p o r t a n tb i o l o g i c arl o l c , t h a t o f a n a g c n ti n r c r v c n i n gi n
t h e m e t a b o l i s mo f l i p i d s .
Chapter2, Note 2. Distribution o[ Potassiumand Sodiunr
A s m e n t i o n e da b o v c , p o t a s s i u mi s t h c c a t i o n o f t h c c y t o p l a s m t, h e
secondarypart of cells, while sodium is the cation of the secondarypart
of the metazoiccompartment,that is of the lluids of this compartment.
A c c o r d i n gt o t h e v i e w p r c s e n t e da b o v e .t h e p e c u l i a rd i s t r i b u t i o no f t h e s c
t w o c a t i o n si n t h e b i o l o g i c a rl e a l m r c s u l t sf r o m t h e i r s i m i l a rd i s t r i b u t i o n
i n t h e e n v i r o n m e n tfsr o m w h i c h t h c s cr e s p e c t i vsec c o n d a r yp a r t s ,c e l l u l a r
a n d m e t a z o i c a, r c c o n s i d e r c dt o h a v c b e e n d e r i v e d .A s w e r e l a t e dt h e
c y t o p l a s mt o m u d , r c s p e c t i v c ltyo t h e l i t h o s p h c r ca. n d t h c f l u i d s o f t h c
m e t a z o i c o m p a r t m e nt to t h e s ea , w e I o o k e df o r a c o n f i r n r a t i oonf t h i s v i e w
i n t h ec o m p a r i s o b
n e t w c c nt h c a m o u n to f t h c s ec a t i o n si n t h c t w o b i o l o g i c a l
c o m p a r t m e n t as n d i n t h e t w o e n v i r o n m e n tw
s h i c h w e c o n s i d e rt o c o r respondto them.
A l t h o u g hp o t a s s i u ma n d s o d i u n ta r c i n a l m o s te q u a l a m o u n t si n t h c
g e n e r acl o n s t i t u t i o o
n f t h e c a r t h ' sc r u s t .p o t a s s i u ni s) f o u n d a l n r o s e
t ntirely
i n t h e s o l i d p a r t sw h i l e s o d i u mf o r n r st h e p r i n c i p a cl o n s t i t u e not f t h e s a l t s
o f t h e f l u i d p a r t o f t h e c a r t h .T h c d i s t r i b u t i o n
of potassium
foundbetween
c c l l s a n d e x t r a c c l l u l afrl u i d ss e e m sv e r y n e a r t o t h a t w h i c h e x i s t sb e t w e e n
l i t h o s p h e raen d h y d r o s p h c r cP. o t a s s i u m
i s f o u n di n a p r o p o r t i o no f 2 . 4 6 0 / o
o f t h e l i t h o s p h e raen d o n l y i n 0 . 0 4 f t o f t h c h y d r o s p h e r(e2 O l ) . T h e r a t i o
o f t h c s er e s p c c t i vceo n c c n t r a t i o ncso r r c s p o n dtso a r i ; , v a l u e .T h i s s e e m s
n e a re n o u g ftro t h e r a t i of o u n di n b i o l o g y W
. h i l et h e e x t r a c e l l u l apro t a s s i u m
r e p r e s e n tosn l y 5 m E q p e r l i t c r , w i t h a t o t a l o f 7 0 m E q ( 2 . 7 g m . ) f o r a
n o r m a l b o d y , t h e i n t r a c e l l u l apr a r t c o r r e s p o n dtso I l 5 m E q p e r l i t e r o f
c c l f s ,w i t h a t o t a lo f 4 . 0 0 0 m E q ( 1 6 0 g m . ) f o r t h e b o d y ( 2 0 2 ) . T h e r a t i o
o f t / r , , f o r t o t a l e x t r a c c l l u l aar n d i n t r a c e l l u l airs a c c e p t e dt o d a y a l t h o u g h
g e n e r a l l cy o n s i d e r etdo o h i g h w h e nc o m p a r c dw i t h t h e p r e v i o u sd a t a g i v e n
y e a r sa g o b y S h o h l ( 2 6 5 ) . T h i s v a l u eo f r . , , a p p c a r si m p r e s s i v e n
l ye a r
t h e r a t i o o f l , rr; f o u n d i n t h e c o m p a r i s o no f t h e p o t a s s i u mc o n t e n to f t h e
lithospherw
e ith that of the hydrosphere.
d h c n c o m p a r i n gt h e p r o p o r t i o n
A s i m i l a rr e s e m b l a n ci es e n c o u n t e r ew
b e t w e e ns o d i u ma n d p o t a s s i u mi n t w o l l u i d s : t h c i n t c r s t i t i afl l u i d o f t h c
. sp p c a rc l o s ee n o u g h .
b o d y a n d o f t h e s e a .T h c t w o r a t i o so f t h e s ee l e m e n t a
g
m
.
h
a
s
1
0
.
4
6
4
o
f
s
o d i u mp c r t h o u s a n d
A
t
l
a
n
t
i
c
Ocean
F o r i n s t a n c et,h e
, h i l e t h e P a c i f i cO c e a n h a s
a n d 0 . 7 2 5 g r . p e r t h o u s a n do f p o t a s s i u mw
1 0 . 2 3 3g r . p e r t h o u s a n do f s o d i u ma n d 0 . 6 3 4 g r . p c r t h o u s a n do f s o d i u n r
( ? 6 6 ) . T h e r a t i o sb e t w e e ns o d i u ma n d p o t a s s i u m
a r e r c s p e c t i v e lly4 a n d
w
h
e
n
t
h
e
a
v e - r a gies c o n s i d c r c da s
i
s
l
6
1 6 , I n t h e b l o o d s e r u m ,t h e r a t i o
548
nESEARcHtN pHysropATHoLocy
3 2 0 m g . o / ao f N a , a n d 2 0 m g . ( / ro f K . T a b l e X X V s h o w st h e s ec o m p a r a tive values.
T n s L EX X V
(a) Comparison
b e t w e e nt h e E x t r a c e l l u l aarn d I n t r a c e l l u l aPr o t a s s i u m
and the amount presentin Hydrosphereand Lithosphere.
Rstio
Potassiumextracellular-total
body 2.7 gm. )
P o t a s s i u mi n t r a c e l l u l a r - r o rbaol d y 1 6 0 g n r . )
Potassiumin the hydrosphere 0.04ch )
Potassiumin the lithosphere
2.46% )
1,59
Ii6l
( b ) P o t a s s i u ma n d S o d i u m i n t h e s e a a n d i n t h e body fluids.
Na
A t l a n t i cO c e a n 1 0 . 4 6 . 1 g r . 0 0
P a c i f i cO c e a n
1 0 . 2 3 3g r . 0 0 0
Blood Serum
320 nrer. 0 0
K
0 . 7 2 5g r . 0 0 0
0 . 6 3 4g r . 0 0 0
2 0 m g r . 0 , ,0
Ratio
14.4
16.0
16.0
T h e s ev a l u e ss e e mt o b i n d t h e d i s t r i b u t i o no f K a n d N a , s e e n b e t w e e n
the cellularand metazoiccompartments,
to that which existsbetweenthc'
environmentsfrom which we considertheserespectivesecondaryparts to
have been derivcd.
Chaptcr 2, Note 3. Social Hierarchic Organization
The organizationof nature with the characteristichierarchic structure
so evidentin the biologicalrealm, has suggested
a similar structurein social
organization.
All the factors which were seento characterizehierarchicorganization
a p p e a rc l e a r l yi n a n a n a l y s i so f s o c i a lo r g a n i z a t i o n( F
. is. 202)The entity,
immediatelyabove that of the individual,is the family. Here, parents and
children-as a grouping of entities of the same level form the principal
part. The secondarypart is made up of elementsof the immediateenvironment, which are kept organizedaround this principal part, and as such are
integratedin the new entity, the family. Housing,goods,even psychological
factors, ideas and habits, characterizethese added factors. A boundary
formation is often much more visible than expected.Living quarters and
comrnon possessionsare well delineated,and charactertznthe farnily. As
expected,most of them are not consideredto belong to an individual but
the family as entity. "This is family property" is a common expression.
Almost always,numerousfamilies are grouped in nearby dwellings.
althoughthis fact alone does not lead to the immediatesuperior entity. the
community.When the group of familiesorganizestogetherand limits certain possessions
taken from the environment,as common to the group.
the entity "community"appears.The principalpart is madeup of the group
of families,the secondaryby the material and even moral goodswhich are
attachedto the goup of families in common. The community has prop-
NorEs
549
lndividualsG
' oods--
Families'Goods
- ---rt.
C o m m ui e
n si t' G o o d s ' C o u ni et s ' G o o d_,/_
s
S t a t e s 'G o o d ts'
i l a t i o n ' sG o o d s
l n t e r n a t i o n aGl o o d s -
In d i v i d u a l s
F a mI i e s
C o n m u inei st
C o u ni et s
- States
N aito n s
H u m aSno c i e t Y
F r c . 2 0 2 . T h e s c , c ' i ohl i e r u r c h i co r g u n i z u ! i o nf o l l o w s t h e s a m e p a t t c r n a s t h e o r g a n i z a t i o n o f m a t t e r o r o f t h e b i o l o g i c a l r e a l m . E a c h e n t i t y r e s u l t sf r o m t h e b o n d o f a
group of lower entities with a secondarypart taken from the environment and limited
by a proper boundary.
proper parts taken from the environmentand common only to this new
entity form the secondarypart. This entity also is defined through its
boundary. It is easy to see how through the same tuerarchic pattern we
passfrom countiesto states,nations,hemispheresand world which reprehigher hierarchicentities.It is interestingto see how, in
sent successively
which we have
each one of thesesocialentities,the samemanifestations
found to characterizethe biologicalentitiesalso exist. The relationship
betweenentities and especiallymany of their functions shows that the
550
RESEARCH IN
PHYSIOPATHOLOCY
social entities are not artificial mental concepts,but are the result of the
interventionof the same forcesin which heterotropicorganizationopposes
the lawlesshomotropy. It has appearedinterestingto seehow much of the
knowledgeof the physiologicaland especiallythe pathologicalmanifestations of the lower entities,we can apply to understandmanifestationsoccurring at the social hierarchiclevels.
Under this aspect,sociologyfinds a new basisnot only for the analysis
of many of its problems,but can have an insight to how nature,through its
own organization,has tried and often succeededin resolving problems.
With the concept of unity in all organization,from subatomic to social
entities,we can understandhow the evolution of the environment,representedby material and intellectualgoods, can produce changesin social
entities.The conceptof highersocialentities,organizedso as to conserve
the characteristicsof the lowest social entities,gives a new aspct to the
relationshipbetween individual, family and society. A scienceof social
physiologycan be createdby systematizinghierarchicsocial entities much
as we did for entitiesin the biologicalrealm. The same approachcan be
applied to social pathology and social therapy as well. Such an approach,
will be the subjectof other presentations.
Chapter 3, Note 1. PrecancerousLesions
Precancerous
lesionswere identifiedespeciallyin casesin which cancerous lesionswere inducedand wherea manifestpolycentricityof lesionswas
present.(203) Polycentriclesionspermit us to study the entiresuccessive
changesfrom normal to invasivecancer.
Induction of cancer in the stomach of rats through carcinogensand
detergents(204) has furnishedexcellentmaterialfor such study; it has
also permitted us to characterizethe specificchanges.Among cells which
appeargrosslynormal, there are somein which certain morphologicalcharacteristicsof the nucleus,notably size and form, appear abnormal. The
existenceof an anomaly is much more evident when the cell divides. lt
may be limited to just a few chromosomeswhich are abnormal in their
dimension and form. This chromosomialabnormality appea.rsstill more
evidentwhen comparedto cellsin mitosisin controlswith normal mucous
membranes.( 309 )
Chapter 3, Note 2. Non-invasiveCancer
We have emphasizedthe characterof the cytoplasm of the cells in
non-invasivecancer,The nuclei show a number of changeswhich, together
a cancerousentity: an irregular shape
rather than separatelycharacterize
of the nucleus with a manifest increasein size; a sharp nuclear border
formed by a dark pigmentednuclear membrane having fine chromatine
particles; a hyperchromatism with clumps of chromatin separated in
bizarre, irregular fashion; and an uneven, irregular distribution of these
chromatin clumps, concentratednear the nuclear membrane. Also often
NOTES
,,, 55I
-552 .'
RESEARCH tN
pHystopATHoLocy
NorEs
553
554
nESEARcH IN
pHysropATHoLocy
is released,the pain that is experiencedduring the period of ischemia disappears,but if the cuff is reinflated,pain may recur without further exercise.In both instances,
no new stimulusis requiredto arousepain. It has
been found that in erythralgia,ncither vasodilatationnor changein skin
temperatureis the factor responsiblefor lowering the pain threshold. According to Lewis, when the circulation to the affectedarea is severelyreduced,accumulatedstablechemicalsubstances
elaboratedby the damaged
tissuesmay act directlyas the pain stimulus.No definiteevidencehas been
offered by these researchers,however, as to the chemical nature of the
elaborated
substances
involved.
2. Local changesin damagedtissuesmay bring about a lowering of
the nerve threshold for pain. Lewis has demonstratedthe spread of the
lowered threshold to nerves far beyond the site of the lesion itself. He
studiedthe cutaneoushyperalgesia
followingtissue-damaglng
excitationof
a tiny area of skin by a tapcredforcepsor faradic current. By producing
damagein a previouslyanaesthetized
area,he found that the local changes
brought about by the damagedid not producehyperalgesiain the surrounding skin until the effectsof the local anaestheticwore off. The localized
nerve changesthen createda wide zone of hyperalgesiafor prolonged
periods.Tower (14) has presented
evidenceto show that the receptorend
structuresfor pain have an arborizingrather than a plexiform arrangement,
thus making unnecessary
the postulationof an autonomicallyunidentified
"nocifensor"nervesystem,as proposedby Lewis, ( l5 ) to accountfor the
type of spreadof the hyperalgesia.
The cxtent and especiallythe distribution of this areaof hyperalgesia
has clearlyindicatedthat it is the result of
a loweredthresholdin the arborizingbranchesof the cutaneousnerve, a
few branchesof which were originally intenselystimulated.When a few
fibers of a cutaneousnerve were directly stimulated,the same effect was
observed.The findings suggestedthat a local tissue change lowers the
thresholdfor pain for the nerveendingsof the damagedarea,and that this
effectmay spreadthrough other branchesof the cutaneousnerve involved
as well as through larger nerve trunks so that the resultantarea of hyperalgesiabecomesvery extensive.
3. Local changesmay alter end organsordinarilyconcernedwith other
forms of sensationin such a way that the impulsesoriginatedby them
evoke the sensationof pain. Certain areassuch as the appendixand the
mucosaof the stomachapparentlycannot,under normal circumstances,
bc
incitedto respondpainfullyto any form of stimulation.(16) However, in
the presenceof inflammation,the same stimuli may give rise to pain in
theseareas.The rclationshipbetweenthe end organsor nervesordinarily
concernedwith the receptionof other forms of sensationand those of pain
has been consideredby severalauthors.Weddell (17) has demonstrated
that the various complex end organ structuresare suppliedwith accessory
fibers,unmyelinatedand beaded.analogousto thoseconsideredto be pain
roceptors.Head ( I 8 ) has shown in experimentson the glans penis that
theremay be a fusionof varioussensations
into a singleconceptand that
one sensationmay inhibit another.Accordingto Feng, (19) the balancein
NOTES
555
556
n E S E A R c Hr N p H Y s l o p A T H o L o c y
storedat 5oC and then brought to room temperature(20"C) before analysis.Fifty cc. of distilledwater was used as a wash when ffask contentswere
transferredto a beaker for titration. The total alkalinity was determined
b y e l e c t r o m e t r itci t r a t i o nt o p H 7 . 0 a g a i n sat . 0 l N h y d r o c h l o r i ca c i d s o l u ,s6
75r
e
75o
?t.
,t6
z' rr.
,t2
,90
/o
c -
cc. KI nzlC0
6t
56
6tt
3:
!3
.-';
6L
60
5l'
_56
Stoo
f,ors
@
::
I fr soto
1 'Y. so2r
60 00
7tl
JrS
too
tf
us
a -
V)o
ner 7
\
67t
2!o
zts 6?5
-----lll-After
6r
50
I
6q
: 6 0
qa
1
s6
5r
O
JQ
t{Irul ts
&
tra
.l!o
Ja l
F t c . 2 0 1 . T h e c o m p a r i s o n b e t w e e nt h e c o n c o m i t a n tc h a n g e ss e c n i n v a r i o u s b l o o d
and urine analysesconccrning lhe ocid-hasebalance of the body. It shows that rhe
variations of the titrimetric alkalinity of the total bloo'd are thc only values which
c o n s t a n t l yp a r a l l e l t h o s e o f t h e u r i n a r y p H . ( P a t i e n t w i t h b r e a s t c a n c e r )
NOTES
557
715
9
o
o
@
?3s
,o cc. fl n/ltlO
78
!
7l
t s
! o <
22
7o
68
66
6e
E .
t9.
(8
< G
'/O
JQr
@
-----'---'------
r l
-t?-
6\'
tJo
60,)
/cp
510
K . L . z
7tl
e
L
a
c
7o
64
rAf{r
6 ^
st
Sa
60'o
- J.
Frc. 204. In some casesthe changes in several blood analysesparallel the cbanges
of urinary pH. (Subject with metastaticmelanoma)
cumulated in the bladder during the half hour precedingbleeding and again
during the half hour following bleeding.The pH values of the urine specimens were plotted separatelyon graphs as A and B curves. Comparison
was then made betweenthe two urine curves and the curve representingthe
valuesof the blood titrimetric alkalinity, using time as abscissa.In another
group of experimentsusing dogs, the bladdercontentswere drained at intervals of from five to ten minutes and blood specimenswere obtained
every thirty minutes.
558
pHysropATHoLocy
. , ' R E , s E A R c Ht N
cc. HCIn/100
oa
3 . .
a
a
I
O
a a
>
'erc
a l
O
(U
_)a
'io
ltl
(u
t
a
(F r n
l n
E
o
o
@
. ?.
a
a
i
a
a
o
a
oa
a
a
ara
o
a l '
a
a
a
aa
U r r n a r yp H
F r c . 2 0 5 . ' I h e c o m p a r i s o nb e t w e e nt h e c o n c o m r t a n tv a l u e s o f u r i n a r y p H a n d o f
t h e t i t r i m e t r i ca l k a l i n i t yo f t h e b l o o d s h o w st h a t t h i s r e l a t i o n s h i pc o n c e r n sm o r e t h e
o c c u r r i n g c h a n g e sa n d l e s s t h e a b s o l u t ev a l u e s o f t h e f i n d i n g s .U r i n e s a m p l e s ' * , i t h
, r e s e e nt o c o r r e s p o n dt o b l o o d s a m p l e sw i t h d i f t h e s a m e p H . i n d i f f e r e n ts u b j e c t s a
ferenttitrimetricalkalinitY.
NOTES
,i
559
cc. C\
4e
947
c,
d
3+a
(lJ
c . -4t
?
J
++
4!
HCIn/100
;c{
3ee
:60
L
?,0
L
? l o
7tl
7,1
7.t
7.2
e
>L
7,1
7.<
D.
Minuteso
f 6rr !'lo
24o
F r c . 2 0 6 . C o m p a r i s o no f t h e h o u r l y b l o o d t i t r a t a b l ea l k a l i n i t y ( i n t e r m s o f c c . o f
. O I N H C I ) a n d h a l f h o u r l y p H c u r v e s i n a h u m a n s u b j e c t .U r i n e c u r v e s A a n d B
r e p r e s e n tt h e b e f o r e a n d a f t e r b l e e d i n g s p e c i m e n s . ' I h e p a r a l l e l i s m b e t w e e n t h e
t i t r a t a b l ea l k a l i n i t y o f b l o o d a n d t h e u r i n e c u r v e A i s c l e a r l y s h o w n , C u r v e B s h o w s
no correlation.
560
nESEARcHrN pHysropArHoLocy
ing the thirty minutes after each bleeding (B curves) did not show the
s a m ec o n s i s t e nct o r r e l a t i o nt o t h c b l o o d t i t r i m e t r i ca l k a l i n i t y .( F i e . 2 0 7 )
In severaltests,collectionsof half-hourly urine specimenswere made
f,fteen minutes before and flfteen minutes after each bleeding.The same
parallelismwas found betweenthc pH of urine accumulatedin the bladder
during the period from fifteenminutesbeforeto fifteenminutesafter bleeding, and the titrimetric alkalinity of blood specimensdrawn in the middle
Elood
cc.
to
9
I
7
6.O
5.8
56
5.4
\---
Hour
-'
F r c . 2 0 7 . C o m p a r i s o n o f t h e c u r v e s r e p r e s e n t i n gh o u r l y b l o o d t i t r a t a b l e a l k a l i n i r l ,
( i n t c r m s o f c c . o f . O l N H C I ) a n d u r i n e p H i n a h u m a n s u b j e c t .U r i n e p H c u r v e
" B e f o r e " s h o w s v a l u e s o f s p e c i m e n sa c c u m u l a t e di n b l a d d e r d u r i n g t h e h a l f h o u r
i m m e d i a t e l yp r e c e d i n ga n d c o l l e c t e da t t h e t i m e o f b l e e d i n g ,w h i l e u r i n e p H c u n ' e
" A f t e r " i s f o r s p e c i m e n sc o l l e c t e do n e h a l f h o u r a f t e r b l e e d i n g .T h e r e i s a definite
p a r a l l e l i s mo n l y b e t w e e nt h e c u r v e so f t h e t i t r a t a b l ea l k a l i n i t y o f b l o o d a n d the urine
pH curve A.
of the urine collectionperiod.The pH curvesof urine specimensaccumulated during the period from fifteen to forty-five minutes after each bleeding showedno consistentcorrelation.
When urine and blood specimenswere obtainedat shorter intervals.
the sametendencyof urine pH changesto precedethe changesin blood
t i t r i m e t r i ca l k a l i n i t yw a s o b s e r v e dI.n F i g u r e2 0 8 a r a p i d r i s e i n u r i n e p H
is seen to begin within twenty minutesof the time of administrationof
sodium bicarbonate.The blood titrimetric alkalinity does not sho,*,an
elevationfor at least forty-five minutes,
NorEs
561
cc
Elood
(.ot N Hct)t7 O
t6.o
r{n
59n..NoHCO,
t,
t40
130
t2.o
9H
Urinc
8.5
8.O
7.5
7.O
6.5
6.O
40
70rc^.90
Minules
ilo
t9
t@
F r c . 2 0 8 . T h e e f f e c to f t h e a d n r i n i s t r a t i oonf 5 g r a m s o f s o d i u n rb i c a r b o n a t eu p o n
the blood titratahle ulkalinitv and urine pH curt'es in dog. The elevation in urine
p H i s s e e n t o p r e c e d eb y t w e n t y - f i v em i n u t e s t h a t o f t h e t i t r a t a b l c a l k a l i n i t y o f t h e
blood.
562
RESEARCH IN
PHYSIOPATHOLOGY
NorEs
563
minor irritant such as a fly or mosquito,for instance-scratching is sufficient to eliminateit. with a fly on his skin, the individualneed not flee or
fight, but only scratch.With the conceptthat itching can result from exactly
the sametype of stimuli as pain and touch,we integrateit in the group of
sensorialsensations.
We can then establisha separatesensorialsubgroup
for itching and pain. whi.le other sensorialsensationsinform us of the
nature of the gxsilsli6n-heat, cold, sound,taste,etg.-itching and pain
inform us only aboutthe intensityof the stimulus,not its nature.
Pathological
itching,like pathological
pain, is relatedto the existenceof
abnormalities,
In addition to the differences
in stimulusintensityrequired
to induceitching and pain, their differentnervousformationshelp explain
their clinical separation.No patient we have studied has ever indicated
any confusionas to whetherhis discomfortwas due to severeitching or
pain. The two sensationsare seldomconcomitant;usually they sucieed
one another.The fact that proteopathicpain and itching both seemto be
conductedthrough unmyelinizednervesindicateswhy they can appear
under similarconditions,as in nerveregcneration.
This semsto have led
to confusionbetweenitching and pain. However, itching and pain observedduring nerve regeneration
can be clearly differentiated
by the patient. The fact that the itching sensationis producedby stimuli of low
intensityalso explainswhy itching is so oftcn presenton skin or mucous
membraneswithout appreciablepathology.Minimal changesappearsufficientto inducethe sensation.
Pain
Touch
It c h i n g
I n t e n s i t yo f e x c i t aito n
Ftc. 208 bis. Similar to pain, itching representsa special kind of sensation,with the
a i m t o i n f o r m a b o u t t h e i n t e n s i t yo f t h e e x c i t a t i o n ,I f t h i s i s v e r y s l i g h t , i t i n d u c e s
i t c h i n g .I f a b o v e t h e t h r e s h o l d ,a t w h i c h t h e i n c i t a t i o na c q u i r e sa n o x i o u s c h a r a c t e r ,
it inducespain.
564
RESEARCH IN
PHYSIOPATHOLOGY
CL
v)
(It
z.
uays
F t c . 2 0 9 . T h e n a s a l p l l m e a s u r e dw i t h a g l a s s e l e c t r o d e i n t r o d u c e d d e e p i n t h e
n o s e s h o w s t h e s a m e d u a l i s m a s t h e o t h e r a n a l y s e sW
. i t h 6 . 5 a s t h e a v e r a g ev a l u e ,
t h c c u r v e so f t h e n a s a lp H h a s m o r e r a p i d a n d b r o a d e r v a r i a t i o n st h a n o t h e r a n a l y s c s .
C u r v e o f d a i l y a n a l y s e ss h o w s v a l u e sa b o v e t h e a v e r a g el i n e i n a c a s e o f g e n e r a l i z e d
melanoma.
7tr
70
o(tt
u,
(u
z.
f 6 7 t
? ro tr (2 rt tr lu r fr /C /9
Days
F r c . 2 1 0 . T h e n a s a lp H s h o w sp e r s i s t e nl to w v a l u e si n a c a s eo f c a n c e ro f t h e l i v e r .
NOTES
565
C h a p t e r4 , N o t e 5 , N a s a l p H
Nasal pH was measuredusing a portableBeckman pH meter and a
glasselectrodesmall enoughto penetratedeepinto the nose.In a research
m a d e w i t h N . B u c h a n a ni t w a s f o u n d t h a t v a l i d d a t a c o u l d b e o b t a i n e d
only if the electrodetouchedthe turbinate,otherwisemarked differencesin
valueswere noted.
7
7
o
ct
(n
(!
z, 6
q
t
Zro
T
(t
lL
6
f
5to
,s
IQ
lt
/3
/1
/6
Uays
b e t w e e nd a i l y c h a n g e so f t h e n a s a la n d u r i n a r y p H s h o r v s
Frc. 2l l. The relationship
oppositevariations.
566
../ RESEARCH tN
pHyslopATHoLocy
NorEs
567
resorption time meant a bad prognosis.A few patients with only values of
2 to 4 minutes died within a few days although other symptomsgave no
indicationof a fatal outcomewithin a short tlmc.
Extendedresorptiontime hasbcenfound in subjcctswith an offbalance
of type A. Values as high as 60 to 90 minutes were found in subjects in whom all other analysesindicated this oftbalance. It is also
interestingto note the existenceof slow resorptiontime for aged subjects.
In a group of 80 patientsrangingfrom 70 to 90 years of age,^n average
resorptiontime of 90 minuteswas found. (FiS. 68)
Chapter4, Note 7. Eosinophiles
The role of the blood as the secondarypart of the entity organismhas
explainedmany of the peculiaritiesof its cells.Aside frorn the phagocytary
functions that can be consideredas a particular form of capturation,the
leucocyteshave to be recognizedas acting as holocrine monocellularformations whose specificconstituentsare liberated by cellular lysis. We have
seen that in the case of the neutrophilicgranulocytes,the hydrolytic enrymes so liberated,stronglyresemblethe externalsecretionof the Dancreas.
Under this aspect,we have investigated
the blood eosinophiles
with a role
similarto the Panethcellsof thc duodenum.
The physiologyof theseleucocytes
has to be soughtin the acidophilic
character of their granules. Morphological analysis of the eosinophile
granule shows that it is formed by a content and a membrane,the last
clearly seenin preparationsin which the granuleshave lost their content.
Like many other membranes,that of the eosinophilegranulecan be easily
identifiedas being made partially at leastby lipids being stainedwith dyes
dissolvingin lipids, such as black Sudan or Scharlach.However, it is the
content of the granule with its ability to combinewith acid dyes that indicatesits specificcharacters.Under certain circumstances,
when the blood
is maintainedin vitro betweenthe slide and cover object for a certain time,
the membraneand granulecontentare seento separate.Before this occurrence,a lysis of the eosinophileleucocyteitself takes place. This is manifested through the breakdown of the cellular membranewith the lysis of
the nucleus.It is in a secondstepthat the eosinophilegranulelosesits content. Following it, besidesthe empty glanulesand lysed eosinophiles,characteristicCharcot-Leydencrystals appear. The correlation betweenthese
crystalsand eosinophileshas been recognizedand is generallyacceptedas
occurringin vivo and in vitro.
A y e r ( 2 1 5 ) h a s s h o r t e n c dt h e p r o c e s so f l y s i s o f t h e e o s i n o p h i l eisn
vitro by treatingthe blood preparationswith a detergent,aerosol.By repeating Ayer's experiments,tle relationshipbetween the appearanceof
the Charcot-Leydencrystalsand the more complex processof lysis of the
eosinophileshas becomeapparent.It could be seenthat the crystalswould
appearat the site where the nuclei of the eosinophilesdisappearedthrough
lysis, and where careful observationof the granulesreveals the loss of
their eosinophilic content. The presenceof empty granule membranes
568
R E S E ^ R c Hr N P H Y S T o P ^ T H o L o c y
NorEs
559
tial condition for their lysis. This situation was apparcnt in a particular
case, that of eosinopeniainduced through administrationof the adrenal
corticoids.Although this appearsto have a direct effect upon the eosinophiles,a definitetime, often even24 hours, is seenessentialin order to
achievethe disappearanceof the eosinophilesfrom the circulating blood.
This delay has been related to the presenceof eosinophiles,allegedly"resistant"to the corticoids.In fact, in studyingthe eosinophileswhich persist
after the administrationof thesehormones,we could see that they represent only immature elements,probably prematurely liberated in larger
amountsfrom the bone marrow. The cells from which some will persist
even for 24 hours after administrationof corticoneoglucogenic
hormones,
do not show lysis in vitro nor the appearanceof Charcot-Leydencrystals,
and they presentthe tinctorial charactersof immaturity for the cytoplasm
and especiallyfor the granules.
Granule maturation,which correspondsto the acidophiliccharacter,
seemsto be the essentialconditionfor the lytic interventionof thesecells.
In the physiologicalrole of eosinophiles,
an importantaspectof maturation was seenin the relationshipbetweenthe richnessof theseelementsin
the circulatingblood and the processesin which a manifest local eosinophilia is induced, as through injection of parasitelarvae or vegetal oils.
A direct relationshipbetweenlocal and sanguineeosinophiliawas apparent, the value of the former being the function of the latter. The ability of
bone marrow to rapidly compensatethe transitory eosinopeniafollowing
the passageof these cells into the tissues,has further directly connected
the local richnessin eosinophilesto the bone marrow's capacity to send
new cells into the circulation.In all thesechanges,the prevailingfactor
has apparentlybeen the degreeof maturity of the eosinophilegranulae,
which seemsto require a certain time to reach thc desired degreewhich
is the principalconditionalso for their physiological
intervention.
Correlationbetweenthe biologicalinterventionof the eosinophiles
and
the acidophiliccharacterof the granulesand their richnessin alkaline
a m i n oa c i d sh a s b e e nc o n f i r m e di n a s t u d yo f t h c b a s o p h i l eosf t h e b l o o d
cells with granuleshaving an oppositecharacter.These granuleshave an
acid content, as seen by their tinctorial affinity for alkaline dyes. They
were also observedto contain heparine.a polysulfonatedmucoid of frank
acid character.The biologicalantagonismbetweenheparineand alkaline
proteinsis well known. Protamines,the correspondentof histonesfor fish,
are usedto correctthe exccsses
of hcparinin thc body. cspeciallyas therapeutic measures.
Therefore,the antagonismbetweeneosinophilicand basophilic granulocytesgoesbeyond their tinctorial characters.
Through the alkaline reactivityof eosinophilesas related to the fundamental separationof interveningconstituentsaccording to their positive
or negativecharacter,the antacid eosinophilescould be consideredto be
in the former group, while the basophiles,
rich in lytic heparin,are in the
latter.
We shall more fully discussbelow the nature of the interventionof the
eosinophilesafter studyingthe role of a specialgroup of constituents.For
570
nESEARcH rN pHysropATHoLocy
E
E
C,
o
C'
u,J
0ays
F t o . 2 1 2 . C u r v e o f b l o o d e o s i n o p h i l e si n a c a s e o f b r e a s t a d e n o c a r c i n o m aw i t h
m u l t i p l e m e t a s t a s e ss,h o w i n g v a l u e sp e r s i s t e n t l yb e l o w t h e a v e r a g el i n e o f 1 0 0 .
In order to understandthis aspectof the eoshophiles,we tried to follow the changesin their amount in the blood in relation to normal and
abnormal physiology.Study of the changesin the number of eosinophiles
in the circulating blood under physiologicalconditions has indicated the
existenceof the same 24-hour oscillationsas seen for many other constituentsof the blood. A relationshipis apparentbetweenthe periodscorrespondingto higher or lower quantiticsof circulating eosinophilesand the
degreeof activity of the individual.This appearsto be oppositein humans
who show diurnal activity, and mice and rats that show nocturnal activity.
By experimentallychangingthe hours of light and dark for mice and rats,
and through it the time of rest and of activity, the rhythm of change was
reversed,
Following the concept of the interventionof eosinophilesin biological
balance,we further investigatedthis aspectof the problem in relation to
the dualism in abnormalconditions.Just as for other tests,we obtained an
NorEs
571
E
(.)
an
o
UJ
Days
F t c . 2 1 3 . C u r v e o f t h e b l o o d e o s i n o p h i l e si n a c a s eo f g e n e r a l i z e dm e l a n o m a s h o w ing values persistentlyabove the averageline of 100.
572
R E S E A R C H
I N
P H Y S T O P A T H O L O G Y
E.
a"!"(:i{llii:','l:":':'."tt'"r'-'r'2trr',",'-i'i'/i','i'i't'i.J.;l'l!l
, . t n { t
,.5
,.4
at
5r,
Ett
2tr
a ,
- . , .
t t 4 , ' 4 ' t t . . t . r
? .
t a n t . ' , 2 , ,
N^
A
.\ rA' \ nA
'r \ VA\ ^ n
l
\
/
\
V
\
^rA
r
V V V ^-J \ i V
\r /
VV
< tt
t
.;
?.'
""'^.-'
)r
C r
6 t
L A
- ' \a A r
lo
J '
t o
I t
tr
jl!
F I c . 2 1 4 . R e l a t i o n s h i pb e t w e e n s e r u m K + a n d t o t a l b l o o d K + prmrts to recogn i z e t h e n a t u r e o f t h e c h a n g e sc o n c e r n i n gt h e i n t e r v e n t i o no f t h i s e l e m e n t . I n a c a s e
of periarteritis nodosa, the bigh values of serum potassium and low values of tbc
total blood potassium indicate an offbalancetype D.
,\!L1t:t4u1Jldt
n /\
i\
V \iV
\
F t c . 2 1 5 . L o w v a l u e so f s e r u m p o t a s s i u ma n d h i g h v a l u e s o f t o t a l b l o o d p o t a s s i u m
i n d i c a t ea n o f f b a l a n c et y p e A i n a c a s eo f c a n c e r o f t h e g a l l b l a d d e r .
NorEs
573
set-up of the apparatus.The blood was diluted with a lvo aceticacid solut i o n i n t h e p i p e t t cu s e df o r c o u n t i n gw h i t c c e l l s .T h e p i p e r t ew a s s h a k e na s
for the count of cells, and the necessaryamount taken from the diluted
content.The potassiumamount was determinedand the result multiplied
by 10. while the averagevalue for the total blood was found to be around
3 8 m E q . ,v a l u e sa s l o w a s 2 0 o r a s h i g h a s 6 0 w e r es c e n (. F i S s . 2 l 4 , 2 l S ,
and 216)
tr4
G
J
60
.ft
7,
on
l l l l | | l | l
(u
a
g
\z
1e
4o
l5
3o
2t
o
o
fo
1o
4z
(g
JE
o
lt
:P9
c
+
Y
F t c . 2 1 6 . L o w v a l u e so f p o t a s s i u mi n s e r u m a n d i n t o t a l b l o o d i n d i c a t ea q u a n t i t a t i v e
d e f i c i e n c y i, n a s u b j e c tw i t h a l i v e r a d e n o c a r c i n o m aT. h e a d m i n i s t r a t i o no f 4 0 m E q
KCI daily, for 9 days brought the two curves to normal.
574
R F . S E A R C } II N
P I I Y S I ( ) T ' AT T I O L O C Y
Poly+rurtrrrtd
Bhlne
alcohols
Aftrr
Aftrr
Beforc
Codliver oll
fatty aclds
Octanol
t
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l
^
LJ\4Jr'*'Lq"/
Aftr
Before
i.^^.^----^**
After
Before
Eeforc
lable
Ursapontf
frrctlon of
Epichlorohydrtn
II
,llvqr.r^.,^.-.*.rrJ
rr.-rb,t+!*L\L
:
Beforc
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h^tn^n",ra"y
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F*l.\fA+,trfl
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t-:
{+^,'--- t ,
Beforc
I : r c . 2 1 6 l J . l i l c c t r o c a r t l i o g n r n r si n. { i r r t l e i r d i n r u b b i t r i n j c c t c d i n t r a p t r i t o n e a l l y w i t h
s u b l e t h ; rdl o s c sr . r fd i l l c r c n t i i g c n l \ . l n t h e g r o u p u ' i t h l i p o i d s * ' i t h p o s i t i v ep o l a r g r o u p s
b e s i d c so t h c r c h a n g c s .i t l l a t t c n i n go f l h c r l a v c T i s i n r l u c e d ,w h i c h c o n t r a s t s w i t h
n m o r e e l e v a t c dT f o r t h e g r o r r p o f l i p o i t h i v i t h n c g a t i v cc h a r a c t e r .
NOTES
5'ts
576 /
n E S E A R c Hr N P H Y S r o p A T H o L o o y
NorEs
5'17
a'L'
A
lf mcasurements
with a capillarytube havinga bore radiusof 0.5 mm.
are madein New York City, whcre the acceleration
of gravity is 981 upon
water which at 18"C has a surfacetension73 dynes/cm.,the heightof the
fluid column is found to be 6.0 cm. and the rclationshipbetweeno and A,
expressedin the cgs. systemis o : 73 A.
A capillary tube thus can be calibratedto permit the direct readingof
the surfacetension in dynes/cm. for any liquid having the same specific
gravity, For fluids of different specificgravity, rhe samecapillary tube can
be usedif a correctionof 0.073 dynes/cm.,is made for each 0.001 increment of the specificgravity.
Urinary specific gravity values encounteredclinically range between
1 . 0 0 1a n d 1 . 0 3 5 ,w i t h a n a v e r a g ev a l u e a r o u n d 1 . 0 1 5 .T u b e s c a l i b r a t e d
to measureurine specimenswith specificgravity valuesat either extreme
can yield errorsin the surfacetensionof as much as 2 dynes/cm.In order
to minimize the degreeof error for routine laboratory use, the capillary
tube has beencalibratedto correspondto a fluid with a specificgravityof
1.015.The maximum error of the surfacetensionvaluesfor the extremes
of specificgravityclinicallyobscrvedwill be reducedto approximately+ I
dyne/cm.in this way. Furthermore,the fact that sodiumchlorideconcentration is one of the importantfactorsinducingdiffcrentvaluesfor urinary
specificgravity reducesthe influenceexercisedby specificgravityupon the
hcightof the column. Sodiumchloriderepresents
a negativesurfaceactive
It will raise the surfacetensionvaluesas its concentrationinsubstance.
e f i t s t c n d c n c yt o n r i u r a t cf r o m t h c s u r f a c ct o w a r dt h e b u l k
c r e a s cbs c c a u s o
of the fluid.This will partiallydecreasc
the influenceexertedby the specific
gravityof the urine, Sincethe surfacetensionvaluesof human urine speci-
578
R E s [ . A R c Ht N p H y s r o p A T H o L o c y
14cm.
NorEs
579
The fluid is drawn slightly above the highestmark by mouth suction and
evacuatedseveraltimes by positive pressure.The tube is again filled to the
same point, care being taken this time that no air bubbles interrupt the
continuity of the fluid column. The tube is removedfrom betweenthe lips,
and the tip of the capillary is then gradually raised toward the surfaceof
the fluid. When the top of the column descendsto the top line (T) of the
scale, the tip of the tube is removed from the fluid and maintained in a
vertical positionat eye level. The descentof the top of the column can best
be observedby viewing the meniscusbetweenthe ends of the split line
calibration markings. The top of the column descendswithin one or two
secondsto an initial point (P1) where it comesto a temporary halt or its
rate of descentsuddenlyslows perceptibly.The column again slowly descends,coming to rest after severalminutesat a secondpoint (P2). After
some time, the descentmay again be resumedat a much slower rate until
a third and final stopping point (P3) is reachedafter more than fi.fteen
minutes.For routine measurements,
the first reading (Pr) is consideredas
the surface tension value of the urine. This correspondsrougNy to the
surface tension value of the specimenbefore any important secondary redistribution of moleculeshas taken place.
The capillary tube should be thoroughly cleaned with distilled water
after use. It is well to check the tube before each seriesof measurements,
using distilled water at room temperature.If the check readingsare above
74 or below 73 on the scale,the tube must be carefully flushed through
with distilledwater by meansof a suctionpump. Occasionally,water alone
may not be srrfficientand it will be necessaryto clean the tube with sulfuricochromiccleaningsolution,followed by thoroughflushingwith water,
in order to obtain correct check readings.when the tube is not in use, it is
best left standingin a gJassbeakercontainingdistilledwater.
Surlace Tension in Clinical and Experimental Research
The Urotensiometerfor the first time makespossibledeterminationsof
the surfacetensionof urine and other physiologicalsolutionsas a routine
laboratory procedure. The highest surface tension value for urine encounteredclinicallyis 73 dynes/cm.,and this is correlatedwith a minimal
quantity of surface-activesubstances.
The lower the surfacetensionof the
urine in dynes/cm.,the greaterthe amount of tensio-activeagentspresent
in the specimen.A surfacetcnsion of 52 dynes/cm. is the lowest clinical
value that we have found by this method in more than 100,000 measurementsmade during the last 12 years.
The first problemconcerningthe meaningof the differentvaluesof urinary surface tension arose when it was observedthat usually the urines
with low specificgravity have high surfacetension,while those with high
specificgravity have low surfacc tcnsion. The direct correlation between
the valuesof surfacetension and specificgravity of the samplesthus had
to be investigatedwith the suppositionthat the amount of water in the
urine will have a great influence,by itself, on surface tension. While a
correlationbetween surface tension and water content is often observed.
580
x E S E ^ R C Ht N p H Y s l o p ^ T H o L o c y
N o r Es
581
:p
olChlqhl r-
I.-
F t c . 2 l E . 2 4 - h o u r s h o u r l y u r i n a r y s u r f a c e t e n s i o n v a l u e o f a 3 O - y e a ro l d n o r m a l
male, kept resting and with a constant hourly food intake, showing a maximum in
the afternoon and a minimum around 5 a.m.
measurements.
During this period, and the entire period of the experiment,
the subjectswere perrnitted to leave their beds each hour to void. Throughout the experiment,they were given the samekind and amountof food each
hour. This eliminatedas variablethe influenceof food and activity.Figures
218,219 and 220 and Table XXVI show samplesof the curvesof surface
tensionin suchcases.A 24-hourdiphasiccurve can be noted.
Surfacetensionin mice undcr similar conditions,however,showsdifterences.A group of 20 mice kept in cageswere used.By slight squeezingof
s82
R E S E A R C H
I N
P H Y S I O P A T H O L O C Y
26 yr mon on stondordrzed
hwrly fad oN lluid rntotc
E
(,
o
c
D
c
o
6
c
e
c,
f
v)
t2
t2
fVoon
lrloon
F t c . 2 1 9 . C u r v e o f t h e u r i n a r y s u r f a c el e n s i o ni n a 2 6 - y e a ro l d m a l e o n s t a n d a r d i z e d
hourly food and fluid intake showinga maximum toward the early morning hours
and a minimum toward the evening.
the lower abdomen,a few drops of urine were obtainedin a little cup and
usedfor surfacetensionmeasuremcnt.
Changesseenin Figure 221 show
that with the passage
of time,thereis a dampeningeffecton the curve.This
has made us doubt that the interventionof strcssin thesecasescan be responsiblefor the changes.In order to eliminatestressas a factor, a second
group of experimentswas done in which urinary sampleswere obtained
rcon
e---
o
D!.
/
o
'----
'
t
----J
a
rta^toht
'
+I.------J
Ftc. 220. Curve of the urinary surface tension in a 27-year old female on stAndard
hourly feeding, with a maximum in the afternoon and a minimum in the morning.
NOrES
,/
583
65
64
:63
E
()
v)
i62
s
;
6l
(l)
()
60
f,
u)
59
58
t2
Noon
g t o 1 2
2 4
I|idnjgh!
t o 1 2
Noon
F t o , 2 2 l . A v e r a g e v a l u e o f s u r f a c el e n s i o n i n t h e u r i n e o f 2 0 m i c e . o b t a i n e d e v e r v
h o u r , s h o w i n g v a r i a t i o n sw i t h a d a m p e n i n gc h a r a c t e r .
584
nESEARcHrN pHysropATHoLocy
!g!r
7 a.m.
8 a.m.
9 a.m.
l0 a.m.
I I a.m.
12 Noon
I p.m.
2 p.m.
3 P.m.
4 p.m.
5 p.m.
6 p.m.
7 p.m.
8 p.m.
9 p.m.
l0 p.m.
I I p.m.
l2 Midnight
I a.m.
2 a.m.
3 a.m.
4 a.m.
5 a.m.
6 a.m.
Male
S.T. in Dynes/cm.
65
64
65
67
69
'70
7l
7l
1t l^
72
73
73
72
70
69
68
67
67
66
66
65
65
63
64
Female
S.T.in Dynes,cm.
66
61
67
69
70
70
72
73
73
72
73
72
7l
7l
'lo
IU
67
67
67
65
65
66
66
65
!
'
'
t. I
,
I
:,r
:.:1': '
i'l
:
NorEs
585
o
o
F.
c
.s
q,
U
o
(,
tz
,\roon
t2
Mtdnght
to
t2
IVoo.t
F t c . 2 2 2 . A v e r a g e h o u r l y v a l u e si n t h c u r i n a r y s u r f a c et e n s i o no f g r o u p s o f 5 m i c e ,
t h c g r o u p b e i n g c h a n g e de a c h h o u r .
586
RESEARCH IN
PHYSIOPATHOLOCY
Becauseof the diurnal patternof surfacetensionvariations,urine specimens were collectedat the same hour every day. During a period of. 22
days, daily urine specimenswere obtained between9 and I I A.M. This
was easily accomplishedby firmly gripping a mouse in one hand by the
scruft of the neck and tail. with the finger of the other hand, the lower
q)
6
a
6
O
{!
aI
(o 6 t
E
l!
a
g
t:
6a
JZ
o
L
:f
o
L
o
E
o
h'
tt
(u
tJ
q
0
o
L
: 99
o
E
o
L
:Y9rn
4)
So
>\
6o
E
E
l
1o
2o
22
2'
UAVS
2t
at
h t c . 2 2 3 . C o m p a r i s o n b e t w e e nw e a t h e r d a t a a n d t h e a v e r a g e v a l u e o f t h e s u r f a c e
t e n s i o ni n 4 0 m a l e r a t s a n d 4 0 f e m a l e r a t s . I t s h o w s a r e l a t i v e p a r a l l e l i s m w i t h t b e
c u r v e o f t h e b a r o m e t r i cp r e s s u r ea n d a m o r e c o n s i s t e n rt e l a t i o n s b i pw i t h t h e i n v c r s e
curve of the temperature.
NorEs
587
F t c . 2 2 4 . A v e r a g ev a l u e o f s u r f a c el e n s i o no f t h e u r i n e i n c o n t r o l m i c e o v e r a p e r i o d
of 3 weeks.
588
RESEARCH IN
PHYSIOPATHOLOOY
41
962
;
o
--
:50
/
Y--
-r'
O
Doys
Frc. 225. Tbc average values of the urinary surface tension of 20 female mice kcpt
in the incubator at 37"c. and of 20 female mice kept in refrigerator at 8"C. The
values are progressivelyincreasing for the animals kept in the incubator until the
a n i m a l d i e d . F o r t h e a n i m a l s k e p t i n r e f r i g e r a t o r ,a f t e r a n i n i t i a l d e s c e n t .t h e v a t u e s
a s c e n dt o w a r d n o r m a l .
l,:-
()
qJ
.cc
t(-r,
at
att
c
/E
-x-/
67
/-l+
Pne-ac'rx.
Cays
Post- adr x.
days
Ftc. 226. Tbe surfacetensionof tbe urine increasesto high values after adrenalcctomv.
5E9
NOTES
r
I
:
'
F
b).
:
Oeltrarg4
c CtL).
0
;
c
lday 7 i
t
l
I
e 6iJ
5 0 tt
Ut
\ I
I
Y
. -,
l
lwv
Oec
1l
2r
'rvn
aC?r
6
Fab
t4
t?
t
il,or
t t
2 t
5 u t 5 2 Q t f ? f
Aq
tloy
Frc. 127. Curvc of the surface lension changes in urine specinrens of a 36-yr.
old prcgnant woman shows a manifest change toward low values, starting with thc
4th month, and becoming especiallylow in the last three months.
I
l4J
(t)
J
q
<t
o
d.9
590
xEsEARcH rN pHyslopATHoLocy
NOTES
59r
rr rt r.,lrr r5 ;7 rl r1
F r c . 2 2 9 . T h e c u r v e o f t h e o x i d o r e d u c t i o np o t e n t i a l o f t h e u r i n e m e a s u r e de l e c t r i cally. The curve of the measurementsmade directly on the urine (El) show big variations which are smaller if the pH of the sample is brought to 7 (E2). In a case of
cancer of thc breast,the curve remains constantly below the 0 valuc.
Frc. 230. Curve of the oxidoreduction potential values of the curve brougbt to pH 7
of a case of cancer of the breast,shows valuesaround or above the 0 value.
592
REsEARcH rN pHysropArHoLocy
cohol was addedto 100 cc, of n/10 hydrochloricacid. 1 cc. of this reagent
was addedto 4 cc. of urine in a test tube kept for a while in boiling water.
The time necessaryfor the discolorationwas marked. values as low as 3-4
secondsor as high as above420 secondswereseen.A high oxidoreduction
potentialinducing a rapid discolorationwas found to correspondto a pattern of the oftbalance typ A while a low discoloration was seen to corre-
C'
(t
-9f
q,
eI
x
o
to
aO
b
F t c . 2 3 l . C u r v e o f u r i n a r y o x i d o r c d u c t i o nv a l u e s i n a c a s e o f c a r c i n o m a o f t h e
breast with multiple bone metastases.The values are establishedas the time ncccssary to obtain the reduction at 100"C and with a pH around 2, of a solution of
toluidinc blue so chosen as to have 100 secondsas the average value for groups of
normal individuals. In this case thc values remain fixed low bclow 100 scconds,correspondingto a patterD,of the offbatancetype A.
spondto the patternof type D. Figures231 and 232 show two suchcurves.
We used this test for many years as main analysesto determinethe exist(220)
ing oftbalances.
Chapter4, Note 14. Peroxidesin the Urine
The hypothesis
of the existence
of a phase"oxygen"of offbalanceD led
us to study the appcarancein urinc of productsrcsultingfrom abnormal
oxidation.We were especiallyinterested
in the existenceof substances
having peroxide properties.We found that addition of sulfuric acid to urine
of certain subjectsinduced appearanceof indigotin and indigo'rubin. In
order to hvestigatethe reaction,we have utilized the solubility of indigotin
and indigo-rubin in neutral solvents.Through their extraction it appears
possiblefirst to preventtheir transformationin colorlessisatin and, second,
NorEs
593
1a
2oo
o
60
C'
f
E
o,
?Y
I
o
-E
>\
x
'@
9o
b
D
h
1o
Days
Fto. 232. The urinary oxidoreduction values in a case of cancer of the colon with
abdominal metastases.The values remain the wholc time above 100 scconds,corrcsponding to the pattern present in the offbalancetype D.
594
xEsEARcH rN pHystopATHoLocy
erysipelasor tonsillitis.In radiation sicknessthe reaction is positive especially when tissuelesionsare manifestsuch as mucositisor epidermitis.In
general,treatmentwith seleniumhas given a relatively high proportion of
positiveresults,especiallyat the beginningof treatment.While positive reaction appearedto be consistentwith a favorableevolution of tumors, an
extremelyintensivereaction appearedrelatedto a bad prognosis.The following observationis characteristic.
o
L
F I c . 2 3 3 . T h e r e a c t i o n f o r p e r o x i d e sr e m a i n s c o n s i s t e n t l yp o s i t i v e i n t h e u r i n e o f a
schizopbrenicin daily analysesduring a period of 3 years. (part of the curve.)
NorEs
595
betweenentitiesand their environmentsunder normal and abnormal conditions. It is under this aspectthat we have investigatedthe renal excretion,
which accordingto the hierarchicorganization,correspondsto the relationship betweenthe organismas an entity and its environment.
some of the substancesexcretedcome from the metabolismof lower
entities.Relatedto blood, they would representoften noxiousundesirable
substances,
if the higher mechanismof the blood would not intervene.Although that which we see as urine is the resultof the relationshipbetween
the organism as an entity and its environment,the origin of the different
substancesforming it, as related to the different other levels, has to be
considered.While for certain elementsthis origin is evident,for many substancesonly suppositionsare availabletoday.
when a systematicanalysisof theseconstituents
was attempted,other
difficultiesarose.The isolatedurine sampleseasily available,have only
a very relativevalue for many of theseinvestigations.
The titrimetricdata
expressedas concentrationof various substancesare all functions of the
amountof watereliminatedin the sample.As this often varieswidely,the
informationsobtainedare only relative.Balanceanalysesconcerningentire
intakesand outputs representsuch technicaldifficultiesas to make them
unavailablefor routine investigation,in which hundredsof subjectsare daily
studied. We tried to bypassthis difficulty by eliminating the factor water
excretion,from the considereddata. The fact that the concentrationof a
substanceand the specificgravity of the urine, are both direct function of
the amount of water present,has permittedto eliminatethis factor. The
ratio betweenthem appearsthus independentof the amount of water
present.It relatesthe amount of a substanceto that of the bulk of the
substances
eliminatedthroughthe kidney.An index of excretionwas thus
obtainedby dividing the concentrationof the substanceby the specific
gravity.The oppositeratio would correspondto an index of retention.
From the physiologicalpoint of view, these ratios are not aftected
by the factors which govern the glomerular filtration, which are acting
present.They are little or not affectedalso
similarlyfor all the substances
by the back resorption,wherethe differences
betweenthe varioussubstances
are reduced.They will show consequently,
big variationsas resultingfrom
which takesplacein the distal portionsof the conthe activereabsorption,
volutedtubes.It is this characterwhich givesthe indexesof excretionor
retention,as we calculatethem, their value. We have utilized for years
theseindexesfor chlorides,sodium, potassium,phosphoricion, sulfhydryl,
calcium, to obtain valuable information which otherwisecould not be
furnishedby the simpleanalysisof the isolatedurinesamples.We will come
back to theseindicesduring further analyses.
Chapter 4, Note 16. Water and Nitrogen Metabolism
The analysesof different urine sampleshave shown that the amount
of water presentin urine seemsto influenceindirectlyits constitution.It
could thus be seenthat while very diluted urine, correspondingto a large
596
nEsEARcH tN pHysropArHoLocy
amount of water excreted, is usually alkaline, concentratedurine, correspondingto small amountsof water excreted,is generallyacid. Furthermore, it could be seen that thesechangesare related to more profound
metabolic differences,When related to the nitrogen metabolism,it could
be seen that diluted urines are rich in free ammonia while concentrated
urinesin uric acid.We havethusinvestigated
the relationshipbetweenthese
two factors,the amount of water excretedand the form under which the
nitrogenis eliminated.
The comparativephysiologyshowsus that the manner under which
nitrogenis excretedvaries for differentanimalsaccordingto the amount
of water availablein the surroundingenvironment.In fish, with water
almostunlimited,nitrogenis excretedin the form of ammonia.The high
toxicityin this form of nitrogenexcretionis counter-balanced
by the amount
of water in which the excretaare diluted.Fish are ammonioselic.In terrestrialmammals,wherethe amountof water availableis more limited, the
excretionof nitrogenis made in the form of urea which is much lesstoxic
than ammonia.The dangerof poisoningthe drinking water throughexcreta
is thus reduced.Mammalsare ureoselicanimals.For birds,for whom water
is scarce,the form of nitrogenexcretionis of uric acid. which through
its low solubility in water, has little chanceto contaminatethe drinking
water.Birds are uricoselic.Basedon this relationshipbetweenwater availability and the type of nitrogenexcretion,we looked for a similar relationship in humansbetweenthe excretableamount of water and the rype of
nitrogenmetabolized
undernormaland abnormalconditions.An immediate
confirmationwas obtainedin thoseabnormalconditionswherethe amount
of water excretedis abnormal.As alreadyseenabove,in subjectshaving
a high diuresis,which in generalwould correspondto a high amount of
water availableto be excretedfrom the body, the urine is usuallyalkaline,
the alkalinitydue to ammonia.On the oppositeside of the normal, there
are subjectswith a very reducedurinary cxcretion.In patientseliminaring
only two to three hundredcc. in 24 hours,the amount of uric acid in the
urine is manifestlyincreased.Upon standing,theseurines alwaysshow a
reddishdepositformed mostly by uric acid. Relatingthis to comparative
physiology,while the normal subjectswould appear ureoselic,those with
polyuria can be consideredas ammonioselicand those with oliguria,
uricoselic.
We tried to see if a changcin the form under which the nitrogen is
eliminatedcan be inducedby changingthe amount of water availableto
be excreted.Normal subjectswhoseurineswere testedfor certain periods
of time for their contentof ammonia,urea and uric acid,were given I to 2
litcrs of water to drink. The highly diluted urine which was subsequently
. h e t o t a l a m o u n to f a m m o n i ai n c r e a s e dw h i l e
e x c r e t e db, e c a m ea l k a l i n e T
urea and uric acid were slightlyreduced.The same subjectswere later
given a dry diet for 12 hours or more. The specificgravity of the urine in
m o s to f t h e s ec a s e sw a s a b o v e1 . 0 2 6 ,i n d i c a t i n ga k i d n e yw i t h n o r m a l c o n d anifestly
c e n t r a t i o cn a p a c i t yA. l t h o u g ht h e c o n t e n it n a m m o n i ad e c r e a s em
in theseurines,the increasein uric acid excretedwas minimal. We sub-
NorEs
59'7
598
.,i
pHysropATHoLocy
RESEARcH rN
7&
778
+ -
776
o
f74
.
- .
- a
- F a - a
- a
- - a
- a - a a a - - . -
- . - . - - - - - - -
7to
78
76
7e4
762
F r c . 7 3 4 . S e c o n dd a y w o u n d c r u s t p H v a l u e sr e m a i n i n a r a n g e f . r o m 7 . 7 2 a n d 7 . 7 6 i n
n o r m a l r a t s a s s e e n i n t h e a n i m a l s w h i c h s e r v e da s c o n t r o l s f o r t h e m u l t i p l e e x p e r i .
ments.
NorEs
599
I n 4 1 0 o f t h e 8 6 0 a n i m a l se x p o s e dt o v a r i o u sc h e m i c a l sp H v a l u eo f
the woundswas determinedseveralminutesafter the operationand found
to be between7.28and 7.35. only 22 showeda deviationfrom the control
range of 7.30 to 7.33. No correlationwas observedbetweenthe minimal
changesin theseanimalsand the type of treatmentadministered.
while the pH values found several minutes after wound induction
showedvery little or no variation from the control rangeand no correlation
with the variousexperimentalconditionsemployed,the findingsat twentyfour hours showedconsiderablesignificancefor the agentsused. At fortyeight hours and thereafter,no important differenceswere observedbetween
valuesfor controlsand thosefor groupstrcatedin differentways. In continuing thesestudiestherefore,determinationswere carried out only several
minutesafter the productionof the experimentalwounds and then again
24 hours later. Actually, only valuesfor the twenty-fourhour readingappearedsignificantand will be discussed.
we made 24 hour measurements
f o r a l l o f t h e 8 6 0 a n i m a l st r c a t e dw i t h d i f l e r e n ta g c n t s F
. or convenience,
we refer to the pH value of the crust at twenty-fourhours after creation
of the cxperimentalwound as the s.d.c.pH (secondday crust pH).
From four to twelve animalswere employedin the assaysof the activity
of each agent.By applyingtie sameexperimental
conditionsto animalsin
groups testedat different times, it was possibleto determinewhether the
changesobservedwere due to some externalfactor such as temperature,
humidity, etc., or were actuallydue to the imposedexperimentalconditions. The s.d.c. pH has proven to be of considerableinterest because
consistentlysimilar changeshave beenfound to be producedby the same
agentswhcn appliedto animalsin differentgroupstcstedweeksor months
apart.
Consideringall the animalstrcatedwith variousagents,three possibilities have been found to exist: I ) There may be no effect upon the s.d.c.
pH, in which casethe valueswill all fall within the control rangeof 7.72
a n d 7 . 7 6 f o u n d i n u n t r e a t e da n i m a l s ;2 ) s . d . c .p H m a y b e e l e v a t e dt o
v a l u e sb e t w e e n7 , 1 7 t o 7 . 8 5 ;o r 3 ) t h e s . d . c .p H m a y b e r e d u c e dt o v a l u e s
of 7.70 to 7.60.
We will presenthere only the conclusionsof thesestudiesas related
to the variousagentsinvestigated.
CHEMICAL
FACTORS
Cationsand Anions
It was interestingto investigatethe influenceexertcdby some cations
and anions by first using the same anion with various cations and then
usingdifferentanionswith the samecation.It was apparentin all the experimentsthat the immediatepH of the wound doesnot differ from that of
the untreatcdanimals,and that the s.d.c.pH data obtainedare concordant.
We studiedthe influenceexertedby anionsfirst by investigatingthc
effectsof admhistrationof acids.With even strong inorganicacids, no
600
R E S E , ^ R c Ht N p H y s r o p A T H o L o G v
NorEs
601
602
nESEARcHrN pHysropATHoLocy
Exlrelad
Lr9@t
786
784
7A2
780
778
775
774
7f?
7 n
- , ^ - _
,o.C .."+il ..*
.fr' urt'itf ,p".
r.
- - -i - l - -J
i
+
.
L.
Lbtotrs,ld
L,pc6
\
*
.d/
F
s{tuat.d
L,@d!
.*." n"*s,
"s
:l
1
,+
!
"C""a I
=-
--
l l
-l-l -1- ---.- l - i rr--.1
F
rtO
na""o
alo
--
c.
_I_.
'
.t --la
r
?''d
768
766
761
762
760
F r c . 2 3 5 . T h e a d m i n i s t r a t i o no f v a r i o u s l i p o a c i d s u p o n t h e s . d . c . p H s h o w s t h a t
while the saturated fatty acids do not influence it, the non-saturatedfatty acids as
w e l l a s t h e l i p o a c i d p r e p a r a t i o n so b t a i n e df r o m d i f t e r e n ts o u r c e si n d u c e a n e l e v a t i o n
of the local pH.
NorEs
60-3
Alcohols
78?
7n
7/8
716
774
77?
774
/a
.tr'
et'
l
(t
+_
tlt
rur
{_
tt
I
cf
((I
;-
''fl
-----|-
76
a
I
764
nn
Ju
"*'*
-;
c q.'c
-j
--
--T-
Co^t6t
I
I
a
a
a
IF
7&
F t c . 2 3 6 . S e c o n dd a y w o u n d c r u s t p H v a l u e sf o r v a r i o u s a l c o h o l ss h o w s t h e r e l a t i o n s h i p b e t w e e na c t i v i t y a n d l i p o i d i c p r o p e r t y .A l l t h e a l c o h o l s b e l o u , b u t a n o l a n d
t e r t . b u t a n o l .w h i c h a r e n o t l i p o i d s ,a r e i n a c t i v eT. h i s a l s o h o l d s t r u e f o r a l c o h o l sw i t h
c h a i n sl o n g e rt h a n e i g h tc a r b o n s .
604
R E s E A R c Hr N p H y s r o p A T H o L o c y
xores
605
Vitamins.
A, D, B.
B ' , B r ,K ' E
Hormones
Testosterone,
Epinephrine
Stilbestrol
Progesterone,
Desoxycorticosterone
Alkaloids . . .
Atropine,
Quinine
Codeine,
Morphine
Antibiotics . .
Penicillin,
Streptomycin,
Aureomycin
Sulfonamides. .
Sulfathiazole,
Sulfamerazine
Antipyretics and
analgesics
Acetophenetidin,
Aminopyrine
Antipyrine
Acetyl-salicylic
acid
Narcotics....
Chloroform.....
Phenobarbital,
Pentobarbital
Liverextract....
A n t i - a n e m i c s. .
X a n t h i n e s . . . . Caffeine
Miscellaneous. . Dicoumerol,
Benzene,
Toluol,
Saponin,
Pteryl-glutamic
acid Teropterin)
Iron (reduced)
Aminophylline
Procaine,
Benadryl,
Coramine,
Glucose,
Glycerine,
Sodium gluconate,
Rutin,
Methyleneblue,
Mercuhydrin,
Benzedrine,
Demerol
606
xESEARcH rN pHystopATHoLocy
7.76
7.74
772
7.68
7.6
7.64
7.62
7.60
7
Ooys
to
d a l k e r t u m o r o n t h c s . d . c .p H d c F r c . 2 3 8 . T h e c f f e c t o f a g r o w i n gt r a n s p l a n t e W
w
o
u
n
d
s
p
r
o
d
u
c
e
d
s
e
r
i
a
l
l
yo n t h e d a y o f t r a n s p l a n ta n d e v e r y
i
n
s
u
r
g
i
c
a
l
terminalions
t
h
e
s . d . c .p H w a s o b s e r v e di n a l l a n i A
m
a
r
k
e
d
r
e
d
u
c
t
i
o
n
i
n
t
h
e
r
e
a
f
t
e
r
.
three days
mals.
NorEs
607
776
7.74
7.72
7.70
7.66
Doys
F r c . 2 3 9 . S e r i a l s . d . c . p H c h a n g e si n a n i m a l s w i t h t r a n s p l a n t e dt u m o r s t h a t r e g r e s s e dr a p i d l y o r f a i l e d t o g r o w , A s l i g h t f a l l i s s e e n t o o c c u r i n t h e f o u r t h a n d
seventh post-transplantday determinations,with a r e t u r n t o t h e c o n t r o l r a n g e i n a l l
the animals by the tenth day.
ffi
-?tLw#--t!lt?l!:aFl
I!+5Effry
608
xESEARCH rN
PHYSToPATHoLocY
BIOLOCICALFACTORS
774
- ---
--
7.72
7.70
7.66
Ooys
F r o . 2 4 0 , T h e s . d . c . p H c h a n g c si n t w o r a t s w i t h g r o w i n g W a l k c r t u m o r s ( - ) .
from which the tumor w8s removed on tbe seventh port-transplant day. Followio3
tho rcmoval, tbc e.d.c. pH began to return towards the control r&n8e. A similar largc
inc.irion was madc in two animals without tumors and no effcct was observed upon
t h e s , d . cp. H i n t b e s ea n i m a l s( , . . . ) .
fourth,seventh,
tenthandthirteenth
post-transpr.",
O"r, ;;;:,J
;::
pH valucsdetermined
eachtime immediately
wound
proafterthe
was
duced,did not differ from the valuesfound in untreatedcontrols. As in all
other anim4ls,the s,d.c,pH only showedsignificantchanges.
775
7.74
7.73
7.7t
4
Doys
F r c . 2 4 1 . S e r i a l s . d . c .p H d e t e r m i n a t i o n isn r a t s w i t h o u t t r a n s p l a n t e dt u m o r s . T h e
scrial small surgical proceduresdid not alter the s.d.c.pH from the control range.
F i g u r e 2 3 8 i l l u s t r a t e sg r a p h i c a l l yt h e c h a n g e si n t h e s . d . c .p H i n a
group of twelve animals with successfultumor transplants.A steady and
maintainedlowering of the s.d.c. pH was observedin each animal, with a
tendencyfor it to rise slightly in thosc animalssurvivingthirteen days.
In some of our animalsthe tumor has shown a tendencyto regress
or fail to take. In six animalsin which the tumor failed to
spontaneously
the s,d.c,pH in the
take or underwentearly regressionand disappeared,
610
xESEARcH rN PHYSToPATHoLocY
fourth and seventh post-transplantday tests fell slightly, but thereafter returnedto control levels.(Fig. 239)
When transplantedtumors that were growing well, were surgically removed following the secondpost-transplantdeterminationon the 7th day,
the s.d.c.pH was seento increaseat the next test.(FiS.2a0)
Surgical wounds. The same incision as for the removal of the tumors
was producedin control animals,but the incision alone did not aftect the
s.d.c.pH. (Fis. 2a0)
In order to further control these experiments,repeateds.d.c. pH determinationswere carried out upon wounds produced serially as in the
tumor bearing animals.When no tumor was present the serial s.d.c. pH
valuesdid not vary from one examinationto the next. (Fis.241)
The serial small surgicalproceduresdid not alter the s.d.c.pH from the
control range.
Chapter 5, Note 2. Potassium
We have seen in Chapter 2, Note I how the place occupied in nature
sodium
and potassiumcan explaintheir peculiardistributionin the body.
by
Proper to the earth crust, one of the environmentsthrough which the
complexindividual has passedduring its phylogeneticevolution, potassium
part of the cellularcompartmentin
appearsas an elementof the secondary
heterotropicelement, potasAs
a
monovalent
the hierarchicorganization.
cation of this compartment.
sium representsthe principal organizational
Its influenceexertedin normal and abnormalphysiologycan be understood
throughits specificinterventionat the cellularlevel, the changesin potassium contentof the other compartmentsof the hierarchicorganizationbeing
sccondaryto thoseoccurringat the cellularlevel.
Potassium,absentat the nuclear compartment,is thus found in the
nuclearor chromosomalsap,only in minimal amounts.We have no direct
information about a passageof potassiumfrom cytoplasminto the nuclear
compartmentunder abnormalconditions.Judging by analogy,it appears
probablethat such a passagewould occur and result in the appearanceof
nuclearvacuoles.
Ample data are availableconcerningthe relationshipbetweenpotassium
of the cellular and of the metazoiccompartments.This information receives
a specialinterpretationwhen relatedto the above mentionedhierarchic
distributionof the elements.
The cells maintain a proPer amount of potassiumin the cytoplasm
which correspondsto a cellularprimary constant.This constantinsuresa
normal cellular metabolismand is controlledin part by the selectiveinterventionof the cellularmcmbranc.Under normal conditions,only a slow
passageof potassiumthrough this membranetakes place as compared to
such as of water, for example.On using radioactive
bther-constituents.
potassium,Moore has shown that it takes about frfteen hours to bring it
into balancewith the intracellularpotassium,while for heavy water, such
an equilibrium was reached in less than two hours. (254) Due to the
NorEs
6ll
612 /
n E S E A R c Hr N p H y s r o p A r H o L o c y
NOTBS
613
614
. , ' R E , s E A R c Hr N p H y s r o p A T H o L o c y
NorEs
615
616
REsEARcH rN pHysropATHoLocy
, ,
)air *
1 , )
?2ai,
due to van der Waals forces (and to the polar groups of A). But we can
simplify this definition.Let us considera solventthe moleculesof which are
dipolar and form polar groupssimilar to thosecontainedin the solute. We
can call those substancesIipoids for which the dipole interaction energy in
the cavity occupiedby the moleculeis lessthan the energyarisingfrom van
der Waals forces.
We get:
* - I,,,,',^""
(o"*,) -' (o"*' * o ) = ) " - e
where Dow o representsvan der Waals forces in a cavity occupied by the
molecule and f the dipolar forces at the surface of the cavity.
Chapter 6, Note 3. Separating Membrane Between Aqueous Media
The possibility of having a membrane with polar groups at both surfaces, as shown in mitochondria, representsone of the most important
meansused in biology to separatetwo aqueoussolutions. (228) One of
the characteristicsof such a membraneis that often it representsan isolating boundary rather than a membranewith functional activity. This is
seen in the fact that very often such a membranebreaks when the two
separatedaqueousmedia have to mix.
Chapter 6, Note 4. Fatry Acids Break Down
In a work hypothesisof the biological breakdown of a long chain
moleculefor caloric metabolism,we considertwo factors as being of capital
importance: 1 ) that the result be an even carbon-numberchain molecule.
since the Knoop oxidation leads to complete caloric utilization only for
suchmolecules,and 2) that the lengthof the chain of carbonsbound to the
carboxyl in the new moleculebe not higher than l1 carbons in order to
permit direct beta oxidation. To fulfill the first condition, the breakdown
processdoes not occur between the carbons of the double bonds themselves,since in natural fatty acids the double bonds separateportions of
the chains,usually with odd number of carbons.Such a brcakdown would
lead to moleculesresultingin further incompleteKnoop oxidation.
According to the hypothesisthe breakdownof the molecule occurs at
the carbon ncarest the double bond. Through the energetic influence
exerted by the double bond, the even numbered carbon nearby appears
strongly positive. As a first step, this carbon was seen to fix a molecule of
NorEs
6l'1
, 7
.
(a)
(b)
\/v A/
\
OoH
(c)
F t o . 2 1 7 . ' f h e o x i d a t i v eb r e a k i n gd o w n o f a f a t t y ' a c i dm o l e c u i e( a ) o c c u r si n v i v o
t a k i n g p l a c et h r o u g h t h e a p p e a r a n c eo f a h y d r o p e r o x i d ea t t h e c a r b o n a d j a c e n tt o t h e
d o u b l e b o n d ( b ) . I t l e a d s u l t i m a t e l yt o a c a r b o x y l f o r m a t i o n ( c ) a t t h i s a d j a c e n t
c a r b o n a o d r e s u l t st h u s i n c h a i n sw i t h e v e n n u m b e r o f c a r b o n s .
If the even numberedcarbon near the double bond is toward the tcrminal methyl group, a monocarboxylicacid will result. A similar process
taking place at the other carbon adjacentto the double bond toward the
carboxyl will lead to a dicarboxylic molecule.The metabolic changesin
vitro-and also i:r vivo-have shown the appearanceof thesetwo groups
of evencarbonmono and dibasicfatty acids.By binding two moleculcsof
water the remaining 2-carbon chain linked by the double bond would re. u c hc h a n g e so c c u r r i n gi n t h e c a l o r i cm o n o s u l t i n a c e t i ca c i d m o l e c u l eS
ethenoidspermitted us to explain one of the baffiing peculiaritiesseen in
the constitutionof the monoethenicfatty acids.
In Note 5, we discussthe positionof the doublebond in the principal
naturallyoccurringmonoethenoids
as it followsa charactcristic
pattern.In
moleculeswith l6 or lesscarbonsthe doublebond is more often placedso
as to separatea group with 9 carbonstoward thc carboxyl end, while in
moleculesof 18 carbonsor more, the doublebond separatesalmost constantlya group of 9 carbonstowardthe methylend. Figure 243 showstwo
characteristicexamples.
t*t
*ooa
t*t
**",
e m p l a c e m c n t sa r c s e e n f o r t h e d o u b l e b o n d i n t w o m o n o Oro. ,or.T*ic
ethenoids.The double bond separatestwo groups with odd number of carbons. For
t h e m y r i s t o l e i ca c i d , i t s e p a r a t e sa g r o u p o f 9 c a r b o n st o l v a r d t h e c a r b o x y l i ce n d . a n d
a g r o u p w i t h a s h o r t 5 c a r b o n c h a i n t o w a r d t h e m e t h y l e n d . F o r h e x a c o s e n o i ca c i d
( 2 6 C ) a c h a i n w i t h l 7 c a r b o n s i s s e p a r a t e dt o w a r d t h e c a r b o x y l e n d a n d o n e o f 9
carbons loward the methvl end.
618
nesEARcH tN pHYsropATHoLocy
,"*AA,/\,/*
i **/y'*'
F t c . 2 4 4 . T h r o u g h t h e e m p l a c e m e n to f t h e d o u b l e b o n d i n t h e m o l e c u l e s o f t h e
m o n o e t h e n o i d st,h e b r e a k i n g d o w n o f t h e m o l e c u l e so f f a t t y a c i d s a t t h e c a r b o n s
adjacent to the double bond leads to the sppearanceof a dicarboxylic acid for the
part of the chain having more than l0 carbons.
i n g t h e c a r b o x y la t i t s e n d , r e s u l t si n a d i a c i d . ( F i 1 . 2 4 4 ) T h i s a l s o w i l l
be subjectto Knoop oxidation,even with a long chain. In the diacid, this
process,which is relatedto the interventionof the carboxyl, can take place
at both endswherecarboxylsare present.
Chapter 6, Note 5. Position of the Double Bond in lUonoethenoids
The comparativeanalysisof the principal naturally occurring monoethenicfatty acids has shown a curiousconfigurationdue to the peculiar
r e l a t i v ep o s i t i o no f t h e d o u b l eb o n d i n t h e s em o l e c u l e sF. i g u r e2 4 5 s h o * s
t h e p o s i t i o no f t h i s d o u b l eb o n d i n t h c p r i n c i p a lm o n o e t h e n o i d s( .2 6 7 )
A n d r e h a ss h o w nt h a t t h e d o u b l eb o n d i s m o r e o f t e n p l a c e ds o a s t o s c p r rate a group of 9 carbonsendingwith the carboxyl.We could show that
this group is presenttoward the extremityending with the carboxyl especially if the chain has l8 carbonsor less.For the longerchain, this group
of 9 carbonsis presentbut usuallytoward the extremity,ending rvith the
methyl. The importanceof this configurationfor the Knoop beta-oxidation
of theseacids is discussedin the previousNote.
NorES
//
619
ic llame
I'lame Systemat
Common
Formula
A 4 , 5 D e c e n o i c *oo"A4AA4'*'
O b t u sI i c
C a p r o l e i c A 9 , 1 0D e c e n o i c xooc&s\AAl"'
A 9 ,l 0 D o d e c e n o i cHoocAAAAI\/,:"
L a i n Ioe i c
A 5 , 6 T e t n a d e c e nxoo<
o i [fuArr1.r41r17c"r
c
o i cn,An.AAA/'n'
l.lyristoleic A 9 ,l 0 T e t r a d e c e n u*
nrlav\'\l'Wvcxs
xooc
P a l m i t o l e i cA 9,l0 Hexadecenoic
ic
P e t r o s ienl i c A 6 r ? O c t a d e c e n o,,*.AAAAAAAA/'''
,.oo.nz\4lA.1yl^yV.cut
0leic
A 9,l0 0ctadecenoic
ic
Vaccenic A l l , l 2 0 c t a d e c e n o-*AA.z,\AAqA/NAIc*r
Gadoleic A 9 , 1 0E i c o s e n o i c ,*"Wt"t
A l l , l 2 E c o s e n o i c eoo(W(*t
C e t r loei c A l i , 1 2 D o c o s e n o i c* o o . W t " t
Eonicic
A i 3 , 1 4D o c o s e n o i c*-.Wc*'
x oiocc W t " l
, 6T e t r a c o s e n o
S e l a c h o l e iAc 1 5 1
A 1 7 , 1 6H e x a c o s e n ou oi ocW t * t
Zr i c o s e n o i c
A 2 1 , 2 'T
Ct{g
F l c . 2 4 - 5 . P r i n c i p a l n a t u r a l l y e x i s t i n g m < l n o e t h e n o i dssh o w t h a t f o r t h e m e m b e r s
w i t h a s h o r t c a r b o n c h a i n t h e d o u b l e b o n d s e p a r a t e sa g r o u p o f 9 c a r b o n s t o w a r d
t h e e n d o f t h e m o l e c u l eh a v i n g t h e c a r b o x y l .F o r f a t t y a c i d s w i t h m o r e t h a n l E c a r bons, the group of 9 carbons separatedis toward the end with the mcthyl 8roup.
620
RESEARCH IN
PHYSIOPATHOLOGY
u.a
a gtz
cta
A rt,n
Cro
+LaJ.
r*- A
Agrz.tt
G1
+a+
t
Czo
qt,t
cto
-f1r
'
tJgtJ-g
L 2,,.,t
czo
,!
L , z .o . ' t
1zr-
,!
ta
22
20
+]Jr
711rJ
Ttro- A
Igt.rt.rt
cao
f i z r o r cr c
Czz
A",z,o.,,
cla
{Js
C
JJs
&t(n'A
l 6 r
A<,0.r;. ,1tt
c zz'
4arL44L41LJJL
S".
t-g
t
J
r
'
to
ta
t'
tL
tt
'o
Ar ro.u,to.,g
czz
H < r a -A
i-:IJ1 l-tO./t /6 /9
czt
,rqJJglJL
2
Lt
6
'gJ
5!.'
to
,LtJ-rJL
/2
/a
s
r.t
JJs
.Ll_
L,+t.to n to
,A
,/ v \,/
c,L4+Lr-Lr!L_rJu!:-!+
)o
2z
)o
22
&-
/a
"
J!qt-
Fro. 246. In the polycthenoids, groups of 9 carbons are recoSnized, formed usuatly by the sum of two or three groups,multiple of 3 (3 or 6). The group of 9 carbons
i s u s u a l l yp l a c e dt o w a r d t h e m e t h y l e n d o f t h e m o l e c u l e .
showed the same iodine number, indicating that in the changeswhich had
taken place, the saturation and desaturationprocesseshad comPensated
each other through a transfer of double bonds from the polyunsaturatedto
the saturatedmembers.
NorEs
621
(+)
(+)
(-)
Linoleicacid
L i n o l e n i ca c i d
(+)
(+)
A r a c h i d o na
i cc i d
F t c . 2 4 7 . R e l a t i o n s h i pb e t w e e n t h e p o s i t i v e c h a r g e o f t h e m e t h y l e n i c c a r b o n a n d
'l-hc
similarity which exists between linoleic
the character of essential fatty acid.
a n d l i n o l e n i c f a t t y a c i d s a s e s s e n t i a lf a t t y a c i d s ,c a n b e e r p l a i n e d b y t h e f a c t t h a t
b o t h h a v e o n l y o n e p o s i t i v e m e t h y l e n i cc a r b o n . A r a c h i d o n i c a c i d , w i t h t w o s u c h
p o s i t i v e l yc h a n g e dc a r b o n s ,h a s t h i s c h a r a c t e ro f e s s e n t i a lf a t t y a c i d s m a r k e d l y i n creased.
This explainsfurther a peculiarrelationshipbetweenthe three important essentialfatty acids. No differencesare seen,from the point of view
of activity as essentialfatty acid, betweenlinoleic and linolenic acid, although the last has 2 intermediarycarbons,one at Crr and another at
C1a.This can be explainedby the fact that the secondintermediarycarbon,
the C1a,as an even carbon, has a negativeelectricalcharacter.From this
point of view of strongly positiventethyleniccarbon, there is no difference between linoleic and linolenic acids. The relationshipbetween the
characterof essentialfatty acid and intermediarypositivecarbon is further
confirmedby the fact that arachidonicacid, with two positiveand one negative intermediarycarbons,is also a more active essentialfatty acid than
linoleic and linolenic acids,each of which has only one positiveintermediary carbon.(FiS. 2a7)
622
xESEARcHtN pHyslopATHoLocy
ctA/"
,rxr[
F t c . 2 4 8 . F i x a t i o n o f h a l o g e n sa t t h e c o n j u g a t e dd o u b l e b o n d s t a k e s p l a c e in t*'o
steps, with a displacementof the double bond in an intermediary position in the
f i r s t p h a s e ,a f a c t w h i c h e x p l a i n st h e n o n r e v e r s i b i l i t yo f t h e p r o c e s s .
NorES
/'
623
differences
noted from one sampleto anotherare so manifestand so consistentthat this mcthod can be consideredas an interestingand reliable
sourceof information.We have,therefore,usedthis techniqueof separation
for thousandsof samplesthroughthe years.
Accordingto the techniquewe devised.the materialto be analyzedtissues,organs, entire organismsor only biological products-is finely
divided in a blender.It is then extractedseveraltimes with ether under
stirring,or in a Soxhletapparatus.Under theseconditions,ether removes
the lipidspresentin the form of free lipids and neutralfats.This represents
a mixtureof the fractionsI + Il. The rcsidueis againextracted,this time
w i t h a m i x t u r eo f l 0 % e t h y l a l c o h o li n e t h e rw h i c h b r c a k st h e c e n a p s e s
a n d s e p a r a t etsh e l i p i d sp r e v i o u s l b
y o u n di n c e n a p s eT. h e r e s u l tr e p r e s e n t s
fractionIII. Thc residucis saponifiedwith l}c/o KOH and extractedwith
ether.This represents
fraction lV US or the unsaponifiable
lipids of fraction IV. After acidification
w i t h t a r t a r i ca c i d . a n e w c x t r a c t i o ni s m a d e
which represents
fractionIV LA or the acid lipids of this fraction.
The ether of fraction I + tl is distilledand the resiductreatedwith
85-90Voalcohol concentrationwhich dissolvesselectivelythe free lipids
as fraction I, and leavesthe part formcd by neutral fats as fraction II.
FractionsI, II and III are then saponifiedseparately.The unsaponifiable fractionsarc extractedwith ether, giving respectivelythe fractions
n i t h t a r t a r i ca c i d , o t h e r e x I U S , I I U S a n d I I I U S . A f t e r a c i d i f i c a t i ow
which
representfractionst LA,
with
made
respectively
ethcr
are
tractions
I I L A a n d I I I L A . E a c h o f t h e s ef r a c t i o n si s w a s h e dw i t h d i s t i l l e dw a t e r ,
dried with anhydroussodiumsulfateand, after the etheris distilledoff, the
fractionsare weighed.
For each material,we obtainedthus four different fractionsfor the
part which we called the US fractions,and four for the
unsaponifiable
saponifiablecalled the LA fractions.They correspondrespcctivelyto:
I , f r e e l i p i d s ; I I , l i p i d s p r e s e n ta s e s t c r so r f a t s ; I I I . l i p i d s b o u n d i n
s h i c h a r e l i b e r a t c do n l y t h r o u g h
c e n a p s eI;V , l i p i d sf i x e di n c o m b i n a t i o nw
In the followingexamples,we chosedifferentmaterialsto
saponification.
illustratethe kind of informationobtainedthrough this method.Fig. 249
the resultsobtainedin normal rats, in rats under abnormalconrepresents
w
d i t i o n sa
. s e l l a s i n t u m o r s ,a t l e x p r e s s eads t h e 8 l i p i d i c f r a c t i o n s .
In order to facilitatethe comparisonbetweennormal and abnormal
c o n d i t i o n sw, e c h o s ef o r t h i s e x a m p l e r, a t s b e t w e e n1 8 0 a n d 2 0 0 g m s ' o f
w e i g h t ,a l l m a l e sw i t h o n e e x c e p t i o n( c a s c b ) . T h e d a t a o b t a i n c dw c r c
thus comparedwith that obtainedfor case (a) which correspondsto a
normal male rat killed by ethcr.Thc analysisfor thc lipoacidfractionsof
t h i sc a s es h o w sa f r a c t i o n( l V ) a t 3 . 5 m g . / p e rg r a mo f a n i m a l ,t h e f r a c t i o n
a t . 2 m g . i p e rg r a m ,l I c o r r c s p o n d i ntgo f a t s a r l ' 2 m g . / p e r
III in cenapse
g r a m a n d t h e f r a c t i o n - la s f r e e l i p i d sa t 8 . 1 m g . / p e rg r a m o f a n i m a l 'F o r
ih. unruponifiablefraction. the fixed part IV is at 0.8 mg./per gram-of
frec
a n i m a l ,t h e I I I i n c e n a p s ca t 2 m g . , t h e I l a s f a t s a t 0 ' 8 m g ' ' a n d t h c
part I at 7 mg./gramof animal.
The female rat (b) showsin general,lower valuesfor the acid frac-
624
RESEARCH IN
PH}'SIOPATHOLOOY
o
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.
c
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q
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b
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*t
g"l
iI
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c
o o
!ts
.!o
! E
. d o
F\
I
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c,o
O E
c
i*-
L]-lls
| , r .t r e e
t:
3E r"
-t{
ut
Frred
rt
o
c)E
a d
--.F\
lE
rr'
f ' r r l i ; . - , 1; n , ; l i t p t , l r
Ftc. 249. Solvent separalion between difrerent lipoacids and unsaponifiable fractions
in various aninrids and tumors. l'hc vlrlucs cxprcssedin mgr,,'grof animal show big
varialions concerning espccialli' the frcc lipidr. (Top fraction)
t i o n sa n c ih i g h c rf o r c u c hf r a c t i o no f t h r u n s i . r p r ) n i R apbl rl ct . I t i s i n t e r e s t i n g
to note that the fixed part (lV) in acid iipids is lorverthan in any male,
'l'hc
a fact conflrnrcd lry otht-.ranalyscs.
increascin unsaponifiablepart
concernsespeciallythe ccnapse"fats and frce lipicl fractions.These differencesare in &grcementwith the data conccrningthe rclationshipbet$recn
lipids in males and females,lrs discussedin Chapter 6.
It is interestingto nc,tethe changr'sin starved animals a$ $hown in casc
NorE s
625
626
x E S E A R C Hr N p H y s r o p A T H o L o c y
The greatconcordance
for all variationshowever,has practicallyoverruled
any objectionof a failure of the separationof thesefree lipidic fractions.
Chapter 6, Note 8B, Spectral Analysis
In collaborationwith Carlos Huesca-Mejiaand PriscillaTeitelbaum.
we studiedseveralthousandsamplesof lipid preparationsthrough spectral
analysis,in ultravioletand first portion of visible light, and more rarely
in infrared,usingthe Beckmanspectrophotometer.
We will limit ourselves
to note here only some of the principalconclusionsreached.
I ) Concerningthc chemicalisomerization
procedures,we could show
the importanceof the temperatureused when a mixture o[ fatty acids is
treated.Thc conjugationin vitro as it is usuallycarriedout, with ethyleneglycol or gJycerolas solvents,was seen to result in preparationswith too
low amountsof tetra-,penta-and hcxacnicconjugatedmembers.A relatively rapid disapparance
of the conjugatedformationswith 4, 5 and 6
doublebonds was seento bc induccdby the high temperaturcused. This
led us to utilizea new methodof conjugation,at lower temperatures.
Using
cthyl alcohol as solvent,prcparationswith high conjugatedformations
were obtained.
2) We utilizedthe spectralanalysisfor quantitativedeterminationnot
only for di-, tri- and tetraencsas usuallyemployed,but also for pentaenes
and hexaenes.For this purposewe determinedthe extinctioncoefficient
to thesepentaeneand hexaeneformations.This was made
corresponding
possibleby isolating the respectivepentaenicand hexaenicconjugated
membersthroughappropriatcsolvents.
3 ) We studied various nraterialsand especiallydifferent organs in
order to corrclatctheir richnessin difTerentfatty acids to thcir biological
activity,by usingthe spectralanalysisof the in-vitro conjugatedfatty acids.
as mentionedabovc.
4 ) S i m i l a r l yw
, c t r i e dt o c o r r c l a t et h e e x i s t e n c o
e f c h a r a c t e r i s t ipce a k s
fractionsof organsto their
in the spectralanalysiscurve of unsaponifiable
biologicalactivity.
5 ) We utilizedspectralanalysisfor the study of the effectsof various
agentssuchas chlorine,sulfur,sulfuricacid or oxygcnupon the conjugated
fatty acids.
6 ) We showedthat minimalchangesare inducedin thenonpolargroups
of conjugatedfatty acidsby changingtheir polar grouPfrom carboxyl into
hydride.
t ith lithium-aluminum
a p r i m a r ya l c o h o l ,b y t r e a t m e nw
7) We studiedthe influenceexertedby conjugatedfatty acids upon
This can be partiallyrevealcdby the quenchingaction induced
carcinogens.
of theselatter agents.
the
fluorescence
upon
8) In an extensivestudy we investigatedthe influenceexerted by
w a sc h a r a c t e r r a d i a t i o nu p o nf a t t ya c i d si n - v i t r oa n d i n - v i v o .T h i s i n f l u e n c e
of conjugatedtrienes,and is presentedin Chapter
ized by the appearance
i
n
" R a d i a t i o n . "a n d o t h e r N o t c s .
NorES
627
(x
$l
o
lt
N
G
9,
't
"r
ti
s'
Llnolenate
o
q1
o
ct
o
CL
l
c
O
t
o
o,
o
cl
0
Llnoleate
o
0leate
Stearate
Palal
toleate
Palmltate
Ityristate
3
t
L i n ol e n a t e
l
o
o
o
r
1
c\
Lrnoleate
Uleate
5teanate
Palnrioleale
i'almii.ate
l,lyrist at e
-,:'
. i
630
R E S E A R C HI N
P H Y S I O P A T H O L O C Y
c
o
E
c
d
L
!l
F r c . 2 5 2 . S p e c t r a l a n a l y s i so f t h e s a m p l e u s e d f o r g a s c h r o m a t o g r a p h y .s h o w n i n
F r c . 2 5 1 , i n d i c a t i n gl h e p r e s e n c eo f d i - , t r i - , t e t r a - ,p e n t a - a n d h e x a e n i cc o n j u g a t e d
members.
NOTES
631
632
REsEARcH rN pHysropATHoLocy
t cHt
(a)
(d)
Ftc. 255. Hypothesis of the synthesis of the allopregnane radicd from arachidonic
acid. Fig. (a) shows the relative position of tbc double bonds in thc arachidonic acid
molecule. Fig. (b) shows how the molecule bends,due to the attraction betwecn tbc
energeticallyoppositely charged C: and C*, C., and C',, and G and C,'. ln Fig. (c)
the cycles are closed with the electronsfurnished by the double bonds. (d) Tbc closing of the cyclopentane occurs with the appearanceof a twin formation. The electrons
of tbe double bond between C, and C, become available at G (G of allopregnan)
to realizc Oe bond with an oxygen at this carbon.
NorBs
633
634
nEsE^RcH tN pHystopATHoLocy
,\^) -.ar--,
lnacttve (a)
Decreasedactrvitv
ll tl G )
Inactrve
t i
\?v
(b)
D e c r e a s e da c t ' " , 1 y G )
./-A/., ")yy (r
?(,u,ty
Decreased
-0x
Inactrve
activrty(jl
Decreased
lr.actrve
(,)
activity
Decreased
- 0
l n aaccti
t r vve
e 0
0))
lnactive
( m)
F r c . 1 5 6 . A n y c h a n g e i n t h e e n e r g e t i cc h a r a c t e r i s t i c so f t h e c e n t e r s w h i c h a p p e a r
r e l a t e d t o t h e s p e c i f i cl u t e o i d a c t , i v i t yl e a d s t o a d e c r e a s eo r d i s a p p e a r a n c eo f t b i s
activity.
NOTES
6]5
636
RESEARcH rN pHysropATHoLocy
dione), would lead to the appearanceof corticosterone(a 4-pregnenell:21-diol-3:20-dione). A further oxidationwould changethis OH into
an O, resulting in dehydrocorticosterone( A pregnene-21-ol-3
: I I :20trione). All theseare mineralocorticoid
compounds.
With a further change, this time at C17,where an hydroxyl would be
attached,all three compounds-desoxycorticosterone,
corticosteroneand
dehydrocorticosterone-displayneoglucogenicproperties. They represent
17 hydroxy derivativesas l7 hydroxy-desoxycorticosterone
(a 4-pregnenel7-9 (beta):21-diol-3:20-dione);17 hydroxycorticosterone
(A 4-pregn e n e - l l ( b e t a ) : 1 7 ( b e t a ) : 2 1 - t r i o l - 3 : 2 O - d i o noer )c, o m p o u n dF ; a n d l 7
hydroxy-1I dehydrocorticosterone
(a 4-pregnene-17:2ldiol-3:I I :20-rrio n e ) , o r c o m p o u n dE o r c o r t i s o n e .
The fact that the presenceof an hydroxyl at Crz greatly changesthe
propertiesof the entire group makesit likely that the energeticformation
of which Crz is a part intervenesin the specificactivity of thesesubstances.
The analysisof the constitutionof the corticoidshas further shown that
they have the charactersof lipoids-being polar-nonpolarsubstanceswith tie nonpolar group predominant,The membersstudied,mineralo- as
well as neoglucogenic
corticoids,have beenshown to induce a changetoward lower valuesin the secondday wound crust pH, indicatingthus a
tendencyto induce an offbalanceof the type A.
Chapter 6, Note 16. The Template Hypothesis
Figure 257 showsthc templateformationin cortisonc,which cxtcnds
from C11to C:r. Each one of thesesix carbonswill attract a carbon from
the radical in front of it. The energeticcharacterof each of the six carbons
of the templatewill determinethc electrophilicor nucleophilic character
of the carbon so attracted.This attraction is easily induced when acetic
radicals,with an clectrophilicand a nucleophiliccarbon,form thesegroups.
Furthermore,the value of the carbonsof the template also will determine
which polar radical will be bound to the respectivecarbon kept in front of
it. In general,the carbon kept in front of a carbon of the templatewill have
an oppositeelectricalsign. Thc polar group bound to the carbon kept in
placewill be oppositein sign to the polar group bound to the carbon of the
template.When the first polar group takesa position parallel to that of the
polar group of the template,both will have the sameelectricalsign.
lt can bc sccnthat C2t within the templatehas an OH, the group being
electrophilic.This will causethe carbon kept in front to preferablybind
an oxygen,realizinga nucleophiliccenter.Czoof the template,which correspondsto a carbonyl, representsa nucleophiliccenter. With an oxygen
bound through a double bond, it has strong reactivity.The positivity of
C3ealso is highly enhancedthrough its bond to the two strongly negative
carbons,C:o and C17,each being bound respectivelyto a hydroxyl. With
this high positivity,C3swill induce a strongreactivity in the carbon kept in
front of it. This will be strongenoughto bind a radical energeticallyopposite to oxygenand strongerthan the hydroxyl;that is, an amino group.
NorEs
637
H2
o=,/'\:->
HL
E,Q
"
O=
o\oH
F r c . 2 5 7 . H y p o t h e t i c a l v i e w o f t h e t e n r p l a t ef o r m a t i o n b e t w e e nc a r b o n l l a n d c a r bon 2l of the corticoid molecule. The groups kept in front of the different carbons
. t h e e x a m p l ep r e s e n t e da b o v e ,
o f t h e f o r m a t i o n s e r v c t o s y n t h e s i z en e w s u b s t a n c e sI n
t h e t e r n p l a t eo f t h e c o r t i s o n em o l e c u l ew o u l d l e a d t o t h e a p p e a r a n c eo f a g l u c o s a m i n e
molecule.
638
pESEARCH rN
pHysropATHoLocy
NOTES
,'
639
Lauric Acid
Myristic Acid
Palmitic Acid
Stearic Acid
A.D.I.
2.5
5
5
1.5
2
t2
6
A.D.I.
6
9
5
A.D.I.
4.5
4
3
20
9
3
50
640
R E S E A R c Hr N p H y s r o p A T H o L o c y
A.D.L of this acid thus showeda 24-fold increaseover the index of its
nonconjugatedisomer.
The data indicatea degreeof specificityfor the adrenaldefensemechanism. A.D.[. va]uesof 3 or lesscould be interpretedto correspondto a
nonspecific
adrenalinterventiontoward fatty acidsin general,while higher
valuesindicatea larger,probablyspecificadrcnal activity. In the case of
alpha-OHlauric and alpha-OHstearicacids,the high A.D.I. valuescould
be releatedto the fact that thesefatty acids have been found to be part of
the constitutionof bacteria.
The intensivedefenseevidenced
by the high A.D.I. valuefor eleostearic
acid is relatedto the appearanceof conjugatedtrienesduring the course of
certainpathologicalconditions,especially
trauma,The continuousincrease
,
especiallyof conjugatedtrienes,in thc adrenalectomizrd
animalssuggests
that thesefatty acids appearin the organismbut are destroyedunder normal conditions.They accumulate,howevcr,in adrenalectomized
anima-ls
and may contributeto deathwhen they reach a certaincritical concentration. It is possiblethat with additionaladministrationof theseconjugated
trienestheir criticalconccntrationwould be reachedand the animalswould
die.
In the light of thesedata, we investigatedthe role of different adrenal
factors in thesc responscs.Cortisone, desoxycorticosteroneacetatc
(DOCA), and sodium chloride were tested for their protective action
againstthe toxic effectsof oleic and eleostearic
acids.Groups of rats were
treatedimmediatelyafter adrenalectomy
with daily dosesof I mg. of cortisone,two-tenthsof a cc. of DOCA, or l7o NaCl drinking water ad libitum.
Control adrenalcctomized
rats wcre given no sustainingtherapy. Three
days after adrenalectomy,
a challengingintraperitonealdose of 1 cc. of
lUVo oleic acid pcr 150 gr. of body weight was administered.
Whereasthe mortality of the control rats was 90Va, it was 25% in
t h c c o r t i s o n et r e a t e da n i m a l sD
. O C A a d m i n i s t r a t i odne c r e a s etdh e m o r t a l ity only to 65Va and NaCl had little effcct,decreasing
it only to 85Vo.
The protectiveeffectagainstoleic acid in adrenalectomy
is seenin thc'
followingtable:
AGENT
C--ontrol
Cortisone
DOCA
NaCl
To MORTALITY
90
25
65
85
T h i s s u g g e s ttsh a t t h c n c o g l u c o g e nhi co r m o n ep l a y s a s i g n i f i c a npt r o tectiverole in the defenseof the organismagainstthe noxious effectsof
fatty acid.The mineralocorticoid,
on the other hand,seemsto play a lesser
role in this mechanism,a fact which is confirmedby the ineflectiveness
of
s o d i u ma d m i n i s t r a t i o n .
A similarpreliminaryexperimentcarriedout for eleostearicacid shorvs
the sameprotectiveeffectiveness
of cortisone,lessereffectiveness
of DOCA
and almost no effect at all of sodium ctrloride.
NOTES
641
642
REsEARcH rN pHysropATHoLocy
NorEs
643
,-0H- [ster
s0,f
'\
sl
Salt
S u l f u r i cA c i d
[)erivatives
(a)
CH
0 - Glycoside
tJH
CH
(JH
CH
\
tH [st ers
CH
CH
CH
c00l-l SaI t
G l u c u r o n iAc c i d
Derivatives
(b)
F t c . 2 5 8 a . T h e b o n d o f s u l f u r i c a c i d i n t h e o r g a n i s mc a n r e s u l t i n a n e s t e r a n d a s a l t .
F I c . 2 5 8 b . T h e b o n d o f g l u c u r o n i ca c i d c a n r e s u l t i n a g l u c o s i d e ,a n e s t e r o r a s a l t
combination.
644
R E S E A R c Hr N p H y s r o p A T H o L o o y
C00H HrN...
hr--Hocc)"'
Ortho
(a)
K- H00C
001-l-H0C
I
H0c
I
H0c
I
fH\,il!
Para
(b)
F t o . 2 5 9 . T h e o r t h o - o x y b e n z o iacc i d ( s a l i c y l i ca c i d ) i s b o u n d i n t h e o r g a n i s m r o
g l y c o c o l l ,( a ) w h i l e t h e p a r a - i s t r m cirr c o u p l e db y t h e g l u c u r o n i cr a d i c a l . ( b ) .
NorEs
645
646
'
R E S E A R c Hr N P H Y s t o P A T H o L o c y
Femolc
Male
tf
lyhole 6roug
n
tJ,
trJ
(t)
-^
> J U
J
li
<l
Tt
o 2 C
|.."
65
k
-hftr*lJlil
66
67
68
59
7,
7t
i:)
73
F t o . 2 6 0 . D i s l r i h u t i o n o f u r i n a r l ' s u r f a c e l e n s i o n v a l u c si n t h e g r < - r uo
pf o l d p e o p l c
studied.
648
x E S E A R C Hr N P H Y S r o p A r H o L o c y
v,
v,
trJ
J
z
o
b.o
fu ond
ovel
65oN
under
Fro. 261. Distribution of surfacetension values in healthy adults and in old age.
s h o w st h e h i g h p r o p o r l i o n o f v a l u e so f 7 0 d y n c / c m a n d a b o v e f o r t h e l a s t g r o u p .
NorEs
649
650
RESEARCH IN
PHYSIOPATHOLOCY
acid (e). Some less marked difterencesfrom the control curve is seen for
the fatty acid preparationobtainedfrom cow spleen(f ).
Independentof these level differences,all the curves of the controls
as well as of the treatedgroupsshow parallel daily changes.An exception
I
to
J
|!
L
f
q)
|J
()
(c,
(c)
='
|J
(J
66
a)
(d)
o
o
o
c
56
50
fll
62
(e)
o - d
L ( I t
> > .
-J+
o
(J
It
..1
r.
V, 6C
g L )
u 6
,t
tt
(f)
JV
i/)
Ftc. 262. Curves of the averagevalue of the urinary surface tension in groups of rats
treated with different fatty acid preparations(l cc of lo/o in oil daily). Thc parallel
changesin the curves, except for oleic acid when the variations are opposit+indicate a common external influence.The differencesin the relationship of the curvs lo
the average value line, corresponds to the direct influence exerted by tbc atsnts.
NorEs
651
652
, , ' R E S E A R c Hr N p H y s r o p A T H o L o c y
sto
L
O
6 l t
Y
!
12
40
t,
o
o
CD
o
o
o
l
6
c
g
L
Jo.26
b!t
,o lo
Joo2
o
e 29
o
o
m
/tu
9o
>
E t o
E
f
-zo
b
G
L
o
75
7o
o
C
o
6o
8/' t
t9
22
21
2{
F t c . 2 6 3 . R e l a t i o n s h i pb e t r v e e nt h e c u r v e so f s e r u m a n d t o t a l b l o o d p o t a s s i u m o f a
g r o u p o f 2 0 s u b j e c t sa n d t h e a t m o s p h e r i ct e m p e r a t u r e .h u m i d i t y a n d b a r o m e t r i c
p r e s s u r eP
. a r a l l e l v a r i a t i o n sa r e s e e nb e t w c e nt h e c u r v e so f t o t a l b l o o d p o t a s s i u ma n d
o f b a r o m e t r i c p r e s s u r e .A n i n v e r s er e l a t i o n s h i pi s s e e n w i t h t h e c u r v e o f h u m i d i t y , .
a n d o n l y p a r t i a l r e l a t i o n s h i pt o t h e t e m p e r a l u r e ,
NorEs
653
temperature,
are broken when a continuouslyunchangedhigh or low temp e r a t u r ei s a p p l i e d .W i t h t h e r e a c t i o n arle s p o n s e x h a u s t e d
a f t e ra c e r t a i n
t i m e . t h e i n f l u e n c e x e r t e da p p e a r sa d i r e c to n e , t h e h i g h t e m p e r a t u r e
ind u c i n ga h i g h u r i n a r ys u r f a c et e n s i o na n d t h e l o w t e m p e r a t u r ae l o w o n e .
However, a differenceis seen betweenthe influenceexerted by the
h i g h a n d l o w t e m p e r a t u r eA. f t e r s o m e t i m e , t h e a n i m a l sk e p t i n a r e frigeratorusuallyrecoverthe capacityto fightthe persistently
low temperat u r e ,a n d t h e s u r f a c et e n s i o nr e t u r n st o n o r m a lv a l u e s T
. h i s d o e sn o t o c c u r
f o r t h e p e r s i s t i n g lhyi g h t e m p e r a t u r eT.h e a n i m a l sd i e w i t h a p r o g r e s s i v e l y
h i c h s u r f a c et e n s i o na s i f t h e d e f e n s i vree s p o n s see e nf o r l o w t e m p e r a t u r e s
w o u l d n o t i n t e r v e n ef o r h i g h t e m p e r a t u r e s .
T h i s d i f f e r e n c ien t h c r e s p o n soc f a n a n i m a lt o w a r da p e r s i s t e n t lhyi g h
o r l o w t e m p e r a t u rcea n b e e x p l a i n e db y t h e f u n d a m e n t adl i f f e r e n c ew h i i h
existsbetweenthesetwo factorsfrom the point of view of homo- or heterot r o p y . w h i l e t h c l o w t e m p c r a t u r eh a s a h o m o t r o p i cc h a r a c t e r t, h e h i g h
t e m p e r a t u rhea sa h e t e r o t r o p iocn e .T h e b o d i c sa r e b a s i c a l l ym o r c p r e p a r e d
t o r e a c ts u c c e s s f u ltl oy w a r dh o m o t r o p i ci n f l u e n c eass t h e y h a v ed o n e i t f o r
m i l l i o n so f y e a r si n t h c p a s t ,t h a n t o a n e n t i r c l yu n u s u a lh e t e r o t r o p i icn flucnce.
Chapter 6, Note 28. Barometric Influence
F o r s e v e r ayl e a r s .w e s t u d i e dt h e p o t a s s i u m
c o n t e n to f b l o o d r e d c e l l s
in subjectswith various abnormal conditions,usually following it daily
for the samepatientfor weeksor months.Besidesother informationswhich
this studyfurnishedand which are discussed
above,we want to emphasize
now the relationshipwith barometricpressurewhich we were able to establishduring repeatedanalyseson the samegroup of subjects.we were
impressed
b y t h e p a r a l l e lv a r i a t i o n ss e e ni n t h e a m o u n t o f p o t a s s i u mi n
t h e t o t a l b l o o do f d i f l e r e n p
t a t i e n t so n d i f f e r e ndt a y sr e g a r d l e sosf m e d i c a t i o n . T h e v a r i a t i o n sc o u l d b e c o r r e l a t e dw i t h c h a n g e si n t h c b a r o m e t r i c
p r c s s u r c sW
. i t h l o w e r p r c s s u r c sa, d c c r e a s ci n t h e a m o u n t o f p o t a s s i u m
i n t o t a l b l o o d ( F i g . 2 6 3 1 w a sn o t c d .L e s sm a n i f e sct h a n g e sw e r e s e e nw i t h
t h c o p p o s i t cv a r i a t i o n si n a t n t o s p h e r ihcu n r i d i t y .
Chapter 6. Note 29. Age, Lipoids and Tumor Transplants
The influenceof ageof a host upon the differentnranifestations
through
the interventionof sterolsand lipoacidscan be scenin the followingexperiments.
Transplantsfrom the samewalker tumor weregraftedat the sametime
i n a n i m a l so f d i f f e r e n ta g e s ,s u c h a s n e w b o r n ,w e a n l i n g sy, o u n g a n i m a l s ,
adultsand aged.The differencebetweentransplants
was evidenteven from
b u t w a s s t i l l m o r ee n h a n c e d
thebeginning
i f f u r t h e rt r a n s p l a n tw
s e r em a d e
i n a n i m a l su n d e r s i m i l a r c o n d i t i o n sA
. ftcr one transplana
t nd especially
after severaltransplants,the followingchangcscould be seen.In the newborn, the tumor took on the aspectof an hemangiomatous
lesion,Therewas
654
nESE^RcH tN pHyslopATHoLocy
no massivetumor and the amount of blood present gave the tumor the
appearance
of a pieceof liver. In weanlings,the characterwas oppositc.
Massive tumors without necrosisand with the aspect of fish meat were
seen.In youngstersthe samecharacterwas obtained.In adult animals,the
tumor had a large portion of necrosiswith predominanceof hemorrhagic
fluid. In very old animals,this characterwas still more accentuated,and
the tumor showedbig cavitieswith hemorrhagicfluid and very little tumor
substancebetween
Similar changeswere obtainedby changingthe site of the graft. By
graftinga portion of the sametumor,'intram-usJularly,
we obtaineda massive whitish,nonnecrotictumor, while subcutaneous
injectioneven in the
samcanimal led to the appearanceof a tumor with multiple nccrotic areas
and caviticsfilled with fluid.
We tried to correlatethesechangeswith the nature of differentlipids
predominantat diflcrent ages.The administrationof fatty acids tcnds to
promotenccrosis,edema,and formationof cavitiesfilled with fluid. while
administrationof insaponifiablefractions,espcciallyfrom placentaor liver,
tendsto produce a type of tumor with whitish, nonnecroticmasses.
Chapter 6, Note 30. Temperature, Lipids and Viral Infection
The relationshipbetweenepidemicsand seasonsis a well-established
concept.In an attemptto explainthis correlation,we considered,as one of
the interveningfactors,the seasonal
changcsin lipids,in view of the influence excrtedby thc two oppositegroupsof lipids upon the receptivity and
manifestations
of infectiousdiseasc.
we
As
havc mentioned,viral infectionsare especiallyinfluencedby the
predominanceof one or the other group of lipids in the body. Among
other factors, temperaturechangeswere found related to this predominance.A relativelydirect correlationwas found for polio, for instance.
The appearanceof neurologicalsymptomssuch as paralysiswas seen to
increaseon days with high tempcrature.
Sucha corrclationcould bc establishedexpcrimentally.Among micc injectedsubcutaneously
with smallpox
v a c c i n ea n d k e p t i n a n i n c u b a t o ra t 3 5 ' C , t h e i n c i d e n c eo f e n c e p h a l i t i s
rose to more than 90% as comparedto 10Vo in controlskept in an airconditionedroom. This correlationrvasfurther explainedby the predominance of sterols in the organism undcr the influenceexerted by high
'l'his
predominance
wasfurther seento inducea higher receptempcrature.
viral
infection,
organism
to
the
and a changein the virus virulence
tivity of
itself, both of which increasewith temperaturcand with the richnessof
sterols.
Chapter6, Note 3l . Youth and Vinrses
g s p e c ot f t h e c h a n g e si n d u c c di n v i r u s e sb y l i p o i d s
A n o t h e ri n t e r c s t i n a
with an alcoholicpolar group,especiallysterols,is the relationshipto age
of the host.Thc greatamountof insaponifiable
fractionpresentin youth-
NOTES
//
655
656
nESEARcHrN pHysropATHoLocy
NOTES
6s7
658
n E S E A R C HI N p H y s r o p A T H o L o c y
NorEs
659
660
RESE^RCH IN
PHYSIOPATHOLocY
NorEs
661
662
, , ' R F - s E A R c r {r N p H y s I o p A T H o L o c y
NorEs
663
Control
lt0
96
ul
4fi
t8
l2
I
8
6
Treated
t2
l9
l8
l5
l0
6
1
tl
5
664
xESEARcH rN pHysropATHoLocy
Trnlr XXIX
Reo Cells SeorpreNr,rtroN
Rrrt'-ntm,i'lst hour
Fatty Acid Used
Stearic
Palmitic
Linoleic
Linolenic
Cod Liver Oil
Control
9
9
9
9
9
Control
53
53
53
Treated
66
54
50
NorEs
665
l a t i o n s h i po f p l a s m at o r e d c e l l sa n d l i p i d s ,T h i s w a s d o n e i n t h e f o l l o w i n g
manner. The cholesterolcontent of red cells was seen to be progressively
lowered by repeatedwashingswith isotonicsaline.when the amount of
cholesterolis reducedbelow a certainlevel,hemolysisensues.Standardizing thesewashingsby replacingthe plasmawith an equalamountof saline,
hemolysisis usuallyobtainedin some bloodsafter I or 2 washingswhile
i n m o s ta f t e rm o r et h a n l 0 w a s h i n g sT.h i s o c c u r sw h e nt h e c h o l e s t e r oclo n -
oc2; &
o Co
(a)
(b)
F t c . 2 6 4 . T ' h e t r e a t m c n to f r e d c e l l s i n v i t r o w i t h c o n j u g a t e df a t t y a c i d s ( t r i e n e s ) i n d u c e st h e a p p e a r a n c eo f v a c u o l e s .( a ) I n s p o n t a n e o u Ise s i o n sc h a r a c t e r i z e db y a n o f f b a l a n c e t y p e D w i t h a p r e d o m i n a n c eo f f a t t y a c i d s o r i n l e s i o n s i n d u c e d b y t h e
a d m i n i s t r a t i o no f c o n j u g a t e dt r i e n i c f a t t y a c i d s , v a c u o l a t e dr e d c e l l s a r e s e c n . ( b )
tent of the red cellsfalls below 58 mgm. Vo. When the separatedplasmais
again added to these repeatedlywashed and consequentlycholesterolimpoverishedblood cells, their cholesterolrises. (Trnle XXXI) By
further additionsof new portionsof the sameplasma,the cholesterolcontent of the red cells can be progressively
elevatedto the original values.
Apparentlycholesterolpassesreadilyfrom the plasmato the red cells.This
Trsle XXXI
I N r . r - u E r c eo r C t r n a r e o P l , r s u l U p o x R E o C e l l C H o l e s r e R o l
BeforeTreatnrent
After l0 Washings
After First Treatnrent
with Plasma
After Second Treatment
with Plasma
186 mgr.9t
62
lil
t42
666
nEsEARcH rN pHysropATHoLocy
226 mgr.7o
96
163
180
NorEs
667
668
nEsEARcHrN pHysropATHoLocy
substances
in thoseclinical conditionsin which similar color changesare
notedin the blood as in shock.We will presentour studiesin shock belou'.
For the momentwe will only note that in the stateof shockexperimentally
inducedby trauma,burns or irradiation,or found in terminallyill adrenalectomized
animals,theseanimalshavenot only a high fatty acid content,
but that the kind of fatty acidsencountered
are not the sameas in normal
animals.We have discussed
theseabnormalfatty acidsabove.The existing
differences
have beenshown by measuringthe quantity of oxalic acid that
is producedwhen thesefatty acidsare submittedto a careful standardized
oxidativefission.The oxidativefissionof the fatty acids not only from their
entirebody but even from their blood has shown that for normal animals,
no oxalic acid could be found, leadingto the assumptionthat no conjugatedfatty acidsare present.On the other hand, oxalic acid appearcdwhen
fatty acidsobtainedfrom animalsin shockor from their blood were broken
down with the analyticalmethod utilized.
Of particularsignificance
for the pathogenicrole of thesefatty acids is
the fact which we will discussagain below, that death appearsto ensue
when the conjugationof fatty acids reachesa certain value, which is ap
proximatelythe same whetherthe animal has been traumatized,burned,
irradiatedor adrenalectomized,
and independentof the fact that death occurs in a short time or severa-l
days.It corresponds
to l4-17 mgm. of oxalic
acid per gram of fatty acids.It is also interestingto note that these abnormal fatty acids were found to be more abundantin the red cells than
in the plasma.
Eflect upon Leucocytes
The biologicalantagonismbetweensterolsand fatty acids has appeared
in the influenceupon other blood constituents.We have observcdthat the
administrationof sterolstends to elevatethe total white blood cell count
and especiallythe number of neutrophilic granulocytes.Polyunsaturated
fatty acids,on the other hand, producerapid leucopenia,and again it is
the neutrophileelementsthat are first affected.A hyperleucocytosis
often
was seenfollowingthe neutropeniainducedby polyunsaturated
fatty acids
if small amountsare administered.This effectcould be consideredas being
reactionalto thc first leucopenia,since it is retardedor even preventedif
large dosesof thesefatty acidsare injected.(Tnnr-e xxxIII)
It is also
interestingto note that a deviationto the right, in Arneth's formula, was
seenafter the treatmentwith fatty acids;and to the left after treatment
with sterols.Thus, this concordswcll with the antagonisticeffectsupon
the agingprocessseenfor theselipids and which is discussedbelow.
Lipids and Blood Serum Cholesterol
Further study of the relationshipbetweenblood and lipids has permitted the recognitionof severalpeculiaritiesconcerningthe blood serum
which when relatedto abnormalconditions,acquiresa specialsignificance.
Policard has observedthat when crystalsof cholesterolare added to blood
sera,two oppositechangescan ensue.In one, a precipitateappearswhile
N O T E S
,,'
669
TrsLE XXXIII
E r r e c r o r L r p r o sA o v t x r s t e R E D
l N V l v o U p o Nr H e T o r , r l N u t ' l s r R
or Leucocytes
Fractionof Blood-lO% Solution-S cc. I.P.
Unsaponifiable
BeforeAdministration
14,600
12,000
2 Hours Later
18,400
19,000
77r Hours Later
26,000
22600
Fractionof Blood-10% Solution-S cc. I.P.
Saponifiable
BeforeAdministration
13,200
16,200
2 Hours Later
I 1.000
6,800
7 V 2 H o u r sL a r e r
5,000
5,100
S t e a r i cA c i d - 1 0 7 o S o l u t i o n - Sc c . I . P .
B e f o r eA d m i n i s t r a t i o n
I6.100
14,200
2 H o u r sL a t e r
12,800
15,100
7 7z Hours Later
I 5,000
I 2,000
the serumcholesterol
contentdecreases.
On the contraryin the other,a part
of the addedcholesterolpassesin solutioninto the serum,therebycausing
an increasein cholesterolcontent. When animals were treated with large
amountsof sterolsfor a long time, their sera showedthis tendencyto precipitationwhen in contactwith cholesterolin vitro, while the sera of animals treated with large amounts of fatty acids showed the capacity of
dissolvingmore cholesterol.We believethat this capacityof sera to precipitate in the presenceof cholesterolmay be correlatedwith the clinical conditions presentin arteriosclerosis
when acute episodesoccur.
Thesestudiesof the role of lipids in blood physiologysuggestthat the
general antagonismbetween sterols and polyunsaturatedfatty acids also
intervenesin other important processes
of blood physiology.It has thus
raisedthe questionof the role theselipids may play, through their opposite
eftects, in different metabolic balancesof the body governed by blood
changes.
Chapter7, Note l. AnalysesUsed for the Study of Hemoshock
Besides body temperature and blood pressure,the following blood
were made: completeblood count;coagulationtime; clot retractaanalyses
bility; values of albumin and globulin, total and free cholesterol.free
fatty acids,degradatedproteins,antitrypticpower of the serum, esterase
amylase,potassium,sodium,calcium and glucose.Most of theseanalyses
were made in venousblood samplesobtaincdevery fivc minutes,during
the fifteenminutesprecedingand the half hour following the noxious intervention.
67t)
RESEARCH IN
PHYSIOPATHOI-OCY
2A
630
subjectssubjects
a
- t q
'.4
lb
(J
wi thout r i t h
i se
exerci se exerc
F t c . l 6 - i . ' I ' h e p r o p r : r t ' t i o no f c h i l l s a p p e a r i n g a f t e r d i r e c t t r a n s f u s i o n ( w i t h t h e
Jubi syringc, injecling 500 cc in less than l0 minutes) increasesmanifestly after cxe r c i s e( w u l k i n g i n t h e r o o m ) .
NorEs
671
.
.1
1' :,.
j.
l.l
, rir:
...1
6'72 .'
R E S E A R c Hr N P H Y S T o P A T H o L o c Y
tf50f.
H-. Control
o-H I nsaponifiablefraction
.-r'| Lipoacids
1 ?3 4 s 6 ? 8 I 1 0 n
D a v s
Flc. 265A. Lipids and ugglutinirres.lnfluence exerted by the administration of unsaponifiable lipid fraction and the acid lipid fraction of human placenta upon the appearanceof ugglutinine.ragainst Eb. typhi vaccinesinjected intravenously. Each curve
c o r r e s p o n d st o t h e a v e r a g ev a l u e o f 5 r a b b i t s .T h e a g g l u t i n i n e sa p p e a r e a r l i e r a n d
t h e i r a m o u n t i n c r e a s e sm o r e r a p i d l y i n t h e a n i m a l s t r e a t e d w i t h l i p i d s t h a n i n t h c
controls.
cod liver oil or butanol or heptanolalso were used.5 cc. of a 2Vo alcoholic
solutionof the lipoids were introducedin I l0 cc. distilled water and rhe
solventeliminatedby boiling the mixture to bring the amount to l0O cc.
l-5 cc. of thesemilky preparationswere added to 5 cc. of the microbe
suspensionand incubatedat 37oC for 24 hours. The treated microbes
were then separatedby centrifugation,the fluid decantedand the microbes
in the same amount of saline.They were used in doses from
resuspended
l/10-l cc. for subcutaneous
injectionsin rabbis, repeatedevery third day
for two weeks.As controls,animalswere injectedwith the same amounrs
of suspensionof untreatedmicrobes.
NorES
7'
673
674
n E S E A R c Hr N p H y s r o r , ^ r H O L o G y
period after the immersion of the patient's hands for ten minutes in icy
water and includedthe followingprocedures:
1) Measurements
of blood pressureand temperatureevery 5 minutes;
2) Determinationsat ten minute intervals,of coagulationtime, clot retraction,white cell count and differential;serum hemoglobincontent,
serumproteins,antitrypticpowerand esterase-allmeasu:ements
being
made on venousblood.
3 ) White cell count and differentialmeasuredon capillary blood obtained
every l0 minutesby finger punctureat 5 rninuteintervalsafter withdrawalsof venoussamples.
4) Tests for the presenceand amount of hemoglobinin the urine at 15minute intervals.
Coagulationtimc was established
in the centrifugetube and was related
to the moment when blood ccasedto flow if the vertical positionof the tube
waschanged.CIot retractionwasdeterminedby centrifugingthe coagulated
blood after 2 hours at room temperatureand measuringthe amount of
serum obtainedfrom l5 cc. of blood. The serum and urine hemoglobin
contentwas determinedphotometrically.
For total proteincontent,we used
both the refractomctricindcx of the serum and gravimetricmeasurements
after adequateprccipitation.For antitryptic power we determinedthe inhibitory effectof the scrum upon the digestionof a solutionof caseinby
trypsin.The quantity of esterasepresentwas determincdby the changes
upon cthyl-butyratc.
The data obtaincdwereplottedas curveswith time as commonabscissa.
Parallelvariationsrvereobservedin all three patientsduring repeatedly
inducedattacks.
For almostall analyses,
exceptfor the presence
of hemoglobinin urine.
t h e v a r i a t i o n si n d i c a t c d: r d i p h a s i cp h c n o m c n o n( .F i g . 2 6 6 ) T h e f i r s t p h a s c
was characterized
clinically by hypotensionand slight hypothermia.The
characteristicanalyticalchangeswere lcucopenia,prolongedcoagrlation
time, reductionin clot retraction,lower refractometricserum value. lou,er
antitrypticserumpowerand increased
scrumesterase.
During this first phase
of the diphasicphenomenon,hemoglobinalso appearedin the serum and.
when abundantin the serum,also was found in the urine. The first phase
was followed by a second5 to l0 minuteslater. The clinical manifestations were a sensationof chill, varyingfrom very slight to severe,followed
by temperatureelevationand slight hypertension.Analytical changesin
the opposite direction from those noted during the first phase could be
seen.Hyperleucocytosis,
reducedcoagulationtime, higher retractionof the
clots, elevatedrefractometricvalue and antitryptic power, and reduced
estcrase
contentwere charactcristic
of the secondphase.Hemoglobinpresphase
ent in the serumin the previous
disappeared
at this time.
The most intercstingfinding in paroxysmalhemoglobinuriawas that
two or three such distinctdiphasicepisodesfollowed each immersion.In
all cases,the first diphasiccomplex appearedin about 10 minutes after
NOrEs
675
Gi -c
:L ( . )
aDt
(n*c
(J
C.a
at)
. o lt
I
+ l
E
5
tG'
vl
eo,
:tc
l- .(uL
.ta
c c
-o.o
+ +
r(f.rr
tlo
lLt
aE
I'linutes
F t c . 2 6 6 . T ' h c c l i n i c a l d a t a a n d a n a l y t i c a lc h a n g e so f h l L x r da n d u r i n c f o l l o w i n g t h e
immersion in icy water clf the hand of a subiect with henroglobinuria a frigore rev e a i t h r e e d i s t i n c t d i p h a s i c p h e n o m e n ac o r r e s p o n d i n gr c s p c c t i v c l yt'o t h r c e s c p a r a t e
hemo shocks. Their intensity appe{irs correlated to the clegree of the occurring
leucopenia.
were followed by the secondphaseof the complex, with scvcrc chill and
manifestchangesin the analyscsin the oppositedirection. lt is the second
phaseof the diphasiccomplcx that is recognizcdclinically as the "attack"
of hemoglobinuria.l'he chill was often very intense,followed by a temperature above 39"C, and hypertension.Leucocytesincreasr:dto as much as
20,000 per cmm., blood coagulationtimc rvas abnormllly shortenedand
clot retractionincreased.Hemoglobindisappearedrapidly from the serunl.
The albumin contcnt as well as the antitryptic porvcr of thc serum increased,whilc the esterasccontent fcll. In about 30 minutes,horvever,all
these changeswere dampened and the blocld slowly rcsained its nornral
This period, with almost all manifestationsslorvlyreturning
characteristics.
to normal often rvas followed by a third diphasicconrplcx.not clinically
evidentbut revealedby hcmatologicalfindings.In most cases,it appcared
about two hours after immersion.While it was much milder than the frrst
two, its diphasiccharacterwas quite clear.Occasionallythc patient rcported
a slight sensationof cold. Thc amountof hemoglobinin the serum was less
676
. . ' R E , s E A R C Ht N p H y s t o p A T t { o t - o c y
I
I
' i, *"t-:
I
/ a !ri)nl
t
io /';"'1
1
I
o
:
O,
J
i
I
,i
f,
\ 1
l
-
l
l
.rl
, ir
* l'
r*1
*.-.J-
{1, .rl
l
F t c . 2 6 7 . I n t h c p a t i c n t w i t h h e m o g l o b i n u r i aa f r i g o r e . e x e r c i s ea f t e r i m m e r s i o n o f
t h e h a n d i n i c y w a t e r i n d u c e sa v i o l e n t c h i l l w i t h a b u n d a n l h e m o g l o b i n u r i a .
NorEs
677
v,
(u
>\
()
o
()
=
q,
t
I
g
o
cl
o
E
|9
CL
R
f
Min:tes
F r c . 2 6 E . T h e a d m i n i s t r a t i o n o f 0 . 0 3 8 m P a n t o p o n p r e v e n t s t h e a p p e a r a n c eo f
m a n i f e s t a t i o n sa f t e r l h e i m m e r s i o n o f t h e h a n d i n i c y w a t e r , i n t h e s u b j e c t w i t h
hemoglobinuria a frigore.
678
,'
RESE^RcH
IN
PHYSIoPATHoLoGy
pathogenesisof the diphasic complex, the influence of morphine has furnisheda meansof preventingthe attacks.
The presenceof two or three successivediphasic phenomena in the
case of paroxystic hemoglobinuriaattack has revealedanother important
relationship.As mentionedabove, hemolysishas been seen to occur only
in the first phaseof the diphasicphenomenon.Hemoglobindisappearsfrom
the serum betweenthe complexes.It is classicallyacceptedthat in cold
hemoglobinuriatwo factors must interveneto produce hemolysis.One is
sensitizationof the red cells which resultsfrom the influenceof cold; the
other is the presenceof the complement.It appearsthat while sensitization
persistsfor a while after exposureto cold, hemolysiswitl occur when the
secondcondition is also fulfilled. Complementis releasedin the first phase
of the diphasicphenomenasinceit is during the first phasethat hemolysis
occurs through the changesinduced by the lysis of the leucocytes.The
correlationbetweenthese two processesis clear from the fact that, under
the influenceof morphine,leucolysisdoes not take place and consequently
hemolysisis prevented.
Chapter 9, Note 2. Lipids and Rouleaux and Sludge Formation
Human blood obtainedthroughvenouspuncturewas mixed in the syri n g e w i t h l / 1 0 o f i t s v o l u m eo f 1 . 5 7 os o d i u mc i t r a t e ,p a s s e di n t o s c v e r a l
centrifugetubes and separatedinto plasmaand cells. The plasma from the
various centrifuge tubes was placed in separatctest tubes. One of these
samplesof plasmawas treatedwith a mixture of conjugatedfatty acids of
cod liver oi.l, another with a lipoacid preparation of human placenta, a
third with a preparationof the insaponifiablefraction of human placenta.
As control, the plasmawas treatedwith liquid paraffin. The mixtures, frequently shaken,were kept in a warm bath at 37'C for one hour, after
which they were centrifugedand the oiJy material separated.The plasma
treatedwith the lipoacidswas then added in proportion of l07o to untreatedplasmaswhich then were reunitedwith their red cells. The plasma
treatedwith the insaponifiablefraction was mixed directly with its respective red cells. Plasma and red cells were shakenfor five minutes and left
at room temperaturefor anotherten minutes.One small drop of this blood
obtained with a platinum loop or a capillary pipette was mixed with two
dropsof salineon a slidecoveredby glassand examinedunder the microscope. While the controls showed few short rouleaux, the blood treated
with insaponifiablefraction showedisolatedcells. In the blood treated with
placenta lipoacids,almost all of the red cells formed rouleaux; in blood
treatedwith conjugatedfatty acid,almostall cellsformed sludges.
Chapter 9, Note 3. Dark Color of the Blood in Shock
In order to determinewhy dark-coloredblood is seenin shock and the
role of fatty acids,the followingexperimentswere performed.
Venousblood of patientsin severestateof shock was drawn and mixed
680
p . E S E A R c Hr N p H y s t o p A T H o L o c y
weight was injected intraperitoneallyor even subcutaneouslyth hour before the animalswere placedin the drum, more than 5OVoof the animals
died during the trauma itself and the fatality rate in some experiments
approached100%. Bleedingfrom the nose and mouth was dark in color
and smallerin quantity than in untreatedanimals.If the same amount of
the lipoacid preparationwas injected immediately after the animal came
out of the drum, it also increasedmortality within the first two hours. For
some animals,death occurredin a few minutesafter the injection.
Chapter 9, Note 7. Infuence of UnsaponiftableFractions Upon
Traumatic Shock
The influenceexertedby the unsaponifiable
fractionsupon traumatic
shockappearedevidentin rats submittedto 5-700 falls in the Collip-Noble
drum. l07o solutionsin sesameoil of the unsaponifiable
fractionof human
or cow placenta,of eggsor of butter,were usedin theseexperiments.I to
5 cc. of thesesolutionswere injectedintraperitoneally
at differentintervals
before or after trauma. The injection of 2 cc., Vz hour before trauma, rvas
seen to entirely preventlethal shock (0/20) in a group of experiments
wherethe mortalityof controlswas 18/20. The sameresultswere obtained
with 2-3 cc. of the preparations
injectedimmediatelyafter the animalswere
takenout of the drum. Dosesas high as 5 cc. injectedone hour after the animal wasremovedfrom the drum, protectedonly a fewanimals(l l/20) and
generallyonly those without symptomsof shock. Once the symptomsof
shock were present,the effectof the unsaponifiable
fractionswas greatly
reduced.(From 2/20 to 5/20 in diffcrentexperiments.)
Chapter 10, Notc I . Oxalic Index
The need to havc quantitativeinformation about the amount of double
bonds presentin the organismor in its lipoacidshas led to a method of
analysisbasedon the fact that molecularbreakdownor fissionwill furnish
characteristiccomponents.With fissionoccurringat the level of the double
bonds,the fraction correspondingto a conjugateddouble bond will appear
as oxalic acid. The problem was to obtain this fissionwith a carboxyl correspondingto each carbon and without having artificially induced displacementsof the doublebond which is a frequentresultof treatment.
We employedthe following technique.Fatty acids from an organism
or any other preparationwere neutralizedwith the exactamountof sodium
carbonatenecessary.
This amount was established
through the neutralization indexof the substances
to be treated.After sufficientdilution,an excess
of sodium carbonatewas added with the aim of obtaining an alkaline
medium.After bringingthe solutionto 4oC potassiumpermanganatewas
addeduntil further discolorationof the permanganate
stopped,after which
ZOVomore of the amount alreadyused was added.The mixture was kept
refrigeratedat 4oC for 16 hours, after which the excessof permanganate
was reducedby sodium bisulfide.The liquid obtained was filtered and the
NorEs
681
Oxalic Acid
nrg gnr of Fatty_Acid
0
tt7
205
| 14.2
l 19.4
99.9
92
u5
80
77.3
40.8
Iodine
Number
180
I19.8
l l5.l
94
96
9l
86
8l,7
76
76.5
57.3
682
RESEARCH IN
PHYSIOPATHOLOGY
T,rsLeXXXV
Ernecr on FoRveltx UpoN rxr, QulNrtry oF Oxllrc Acro Pnoouce,oBy
Oxro,rtrve Ftssrox on A Mrxrune on CoN.luc,rrep F,rrry Acns. (5 cc.
Alrquors or A MrxruRL, oF CoN.luclrro F,rrry Acros Wenr, MxEo Ix A
Sropprnrp Cyr-rxorn Wrrn l0 cc. oF A 20Vo Fonvol Solurlox. THE CyLrNoen Wrs Snlxen ,rNn S,rprpr-rsWrnr Rrproveo Ar FneeuENT INTERVATJ
Fon DerenMrNArroNon Oxruc Acro.)
Time
Before mixing
After
I hour
"
24 hours
"
48 hours
"
72 hours
"
144 hours
tzt.7
121.8
120.5
120.9
I 18.9
t2t.9
NorEs
683
ir-1;-_z-r7
:---'wr==^.<r'i;-zv-Ul^.
\,/-
684
c , E s E A R c Ht N p H y s r o p A T H o L o c y
_-^---^
. -Vf
.,^t--V\--:
1r.\
F t c . 2 7 l . K . L . 5 2 y e a r so l d . C a n c e ro f t h e h y p o p h a r y n xT. r e a t e dw i t h 3 0 0 r f o r e a c h
s e s s i o n .T h e h i g h s p e c i f i cg r a v i t y s h o w s a t e n d e n c y l o w a r d t h e t y p e A . a m o n t h
a f t e r t h e c o m p l e t i o no f t h e t r e a t m e n t .
W h i l e s p c c i f i cg r a v i t y a n d S T s h o w a s l i g h t o f f b a l a n c c[ ) d u r i n g t h e t r e a t m e n ta n d
t h e p H a m a n i f e s tc h a n g et o w a r d t h e t y p e D , f o l l o w i n g t h e r a d i a t i o n a l l t h e a n a l y s e s
show a changetoward the type A, one month after completion of rhe radiation.
P e r o x i d e sp e r s i s t e di n t h e u r i n e . C l i n i c a l l y t h e e n t i r e e v o l u t i o n w a s e v e n t l e s s .
.:JJ.
J-^v
L .r .
"iF:
l
-/Y
^ L : .
u t t ; -
f---\'-^r
\-
\2---=^-f
zr'r
al'.1
la
8
len0x.
F r c . 2 7 2 . L . W . , 5 8 y e a r s o l d . C a n c e r o f t h e l u n g . A f t e r r e c e i v i n go n l y 1 0 0 0 r ,v c r y
w e a k . M u c h w o r s e a f t e r , w i t h v e r y r a p i d d o w n h i l l c o u r s e .A f t e r t b e t r e a t m e n t o n
3 / 5 , t h e p a t i e n t e n t e r e di n t o c o m a a n d d i e d 3 / 7 . T h e e x t r e m e l yh i g h v a l u e s f o r C l I
a n d h i g h s p e c i f i cg r a v i t y c h a r a c t e r i z et h e a n a l y t i c a lc h a n g e s .I t i s i n t e r e s t i n gt o n o t e
tbe negative reaction for peroxidesfor almost the entire time.
IiIIEiI
ESEEES
i:
E5$ E
I:;Ei!F
l = &E ; g ;
I E fr U;T 5
i : 3gE ;
s : ; E!:
E 8B : EFE
H Ers5s s
l6 c: I : i 0
F . E . :E t
i;r!r::
;igEigfi
NorE s
697
1iJ0r.
F r c . 2 7 4 . C a n c e r o f t h e c e r v i x i n t e n s i v e l yi r r a d i a t e d . A l t h o u g h c h a n g e s o c c u r i n
the analyses,the p""ssges in offbalance D do not coincide in the difterent tests. With
the continuation of the treatment, the patient passcs in offbalance A. No clinical,
noxious manifestations were seen.
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lJr
NOTES
689
F t c . 2 7 6 . F . 8 . , 6 4 y e a r s o l d . C a n c e r o f t h e f u n d u s o f t h e u t e r u s ,t r e a t e d w i t h 4 5
mgr. radium for 122 hours in situ. with a total of 5490 mgr. hours.Felt subjectively
w e l l a f t e r r a d i a t i o n w i t h o u t a n y c o m p l a i n t .T h e a n a l y s e ss h o w a m a n i f e s t c h a n g ef o r
almost one week from the type A to type D. For the S.T. it starts 2 days after the
i n s e r t i o no f r a d i u m w i t h t h e v a l u e sd r o p p i n g f r o m 7 0 t o 5 5 . F o r t h e p H , t h e c h a n g e
s t a r t e da f e w d a y s l a t e r w i t h t h e v a l u e sp a s s i n gf r o m 7 . 8 t o 4 . 6 . T h e c b a n g e so f C l I
s h o w v a l u e sa s h i g h a s 7 w i t h o u t h o w e v e r ,h a v i n g t h e c h a n g e sc o i n c i d ew i t h t h e p H
a n d S . T . t 6 d a y s a f t e r r a d i u m w a s t a k e n o u t , t h e a n a l y s e sw e n t t o p r e v i o u s v a l u e s ,
cxcept the peroxides,which remain present.
690
nesEARcH rN PHYSIOPATHOLOGY
0
I
NHr
I
C - S l+ H 0 - C - C - R +
t
ltl2
-C-M
HNA
groupleadsto the
Frc. 277.The bondbetween
acidandan amino-acid
thecarbamic
group.
formation
of theNCNC
genicactivityof urethanetakesplace at the lowestlevelsof organization,
explainsthe necessitythat a certain time separatesits interventionfrom
that of croton oil, which would act only at the higher levels,probably
inducing the passagefrom noninvasiveto invasivephase. This timc is
necessaryfor the first cancerouschangesto build up the seriesof cancerous
hierarchicentitiessincc the cocarcinogcn,
croton oil, would act only in
those more evoluted cancerousentities.In experimentsin course, the
passageof the noninvasiveurethane-induced
carcinogenesis
entitics into
invasivecancer,is successfully
inducedby preparationsof unsaponifiable
fractions.
Chapter 11, Note 2. Constitution of Viruses
It is superfluous
to emphasize
the interestwith which virusesare being
studiedfrom all points of view. Their role in carcinogenesis
has placed
them in the limelightof cancerresearch,and any contributionconcerning
their constitutionor activity is of great interest.A much debatedinitial
problem concernsthe nature of viruscsand their place among the other
entities.(293)
Two fundamentalgroupsof constituents-DNA and proteins-(301.
289) havebeenrecognized
to takepart in theformationof theviruses.These
two groups could be separatedand reunited, reproducingthe original
virus with all its characters.Futhermore,new virusescould be created
when fractionsresultingfrom differentviruseswere bound together.(289)
However,the fact that a part of the virus, the DNA fraction,was seento
be furnishedby the constituents
of the host cell, and the protein directly
by it ( 312), has raisedthe <;uestion
of the natureof the virus itself. Some
workers have gone so far as to seethe virusesas parts of the constituents
of the cells. By consideringthe virusesin the conceptof the hierarchic
organizationpresentedabovc, a new aspectemerges.
Accordingto this organizational
concept,a virus representsan entity
that has reacheda certainstep in the hierarchicevolution,and remained
Like all entities,a virus can be contherethroughoutits individualization.
ceivedto be formed by a principal part bound to a secondarypart. the
ensuingentity limited by a boundaryformation.The principal part would
be formed by a group of immediatelyinferior entities in the hierarchic
NorES
691
692
R I:Sl:ARC ll
I N
;S,tS.
"iil
aD.
I
- ?
Sr
Fu; l l l { . I r t i e r r c t r o no f h u r n a ns e r u n tu p ( ) nS u 1 t i Oa s c i t c sc c l l s .
{I)
t1t
(3l
( : 1)
[ - \)
( ( r)
. llli
693
NOTES
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(9)
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it
Con t inuatic'rn-'.
N o r m a l S A l l { 0 ( c l l r X l 8 { ) 0 .( i i e n r r i t \ t i l n .
C a r c i n o l y ' t rcct l c c t o f S e r u mo n S , { c c l l s X l t { 0 1 }(.i i c r n s l r t ; r i n .
N o r r n a l l - 1 r i s o f S , \ l E { ) ( e l l r ; r s c o n l r i . r il n P o l r r i r c c h . i r r d el ns h r b r t i o n S t u d i e so n C ' l r c r n o l ir t s , X . - l ( X )n. t r t s l i i i n e d .
( l 0 ) E l l c c l o n C ' a r c i n o ls\ i r h y t h e P o l l : a c c h a r i d eD e. ' t t r a nX 1 0 0 , n o t * t a i n e d .
i l l r E f f e c t o f l - c v a n o n ( a r c i n o l v s i r .N t ; t e t h c c l c a r l y d i s t i n c t o u l l i n c o f u n a f f e c t e dc e l l s . X l ( X ) . n { ) t r t a i n e d .
i l l ) E f f e c l o f i l u c i n o n ( - u r c i n o l y r i rX 3 0 0 , n o t s t i i i n c d ( e v i d e n c eo f i n h i h i t e d
Lysis.
694
s , E s E A R c Hr N p H y s r o p A T H o L o c y
NorEs
695
696
x E S E A R C HI N P H Y s r o P A T H o L o c y
The fluorescence
of carcinogenichydrocarbonsis measuredby meansof
the fluorcscentattachmentto the Beckman spectrophotometer
using a
wavelengtharound365 mp. The sensitivityof the apparatusis adjustedto
show a value of 100 for the fluorescentlight utilizing the concentrationof
the hydrocarbonhaving the maximum fluorescence.
The carcinogenichydrocarbonsare dissolvedin alcohol,iso-octans
or cyclohexane,
the last two
being purified and thus renderedoptically inactive by passagethrough a
silicacolumn.
The fatty acidsare dissolvedin varyingdilutionsin the samesolvents
and addedto the solutionof hydrocarboncarcinogenwhich has been previously chosento give a fluorescentvalue of 100. The fluorescenceof the
mixture is immediatelydetermined.The quenchingeflect of the fatty acid
is shownby the percentof the residualfluorescence
of the carcinogenwhen
mixed with differentsolutionsof a mixture of conjugatedfish oil fatty acids.
Tesle xxxvl
showsthe quenchingeffectof mixturesof fatty acidsconjugatedby treatmentwith KoH upon differcntcarcinogenicand related
hydrocarbons.
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lave Lemth!rt)
tb
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F t c . 2 8 0 . C ' o n j u g a t i o nm e t h o d .
use of ethylic alcohol as solventfor conjugation
c h a n g e st h e a m o u n t o f p o l y c o n j u g a t e dm e m b e r s o b t a i n e d f o r c o d liver oil fatty
a c i d s ( a ) a s c o m p a r e d w i t h t h o s e o b t a i n e dw h e n t h e c o n j u g a t i o ni s made at higbcr
t e m p e r a t u r e su s i n g e t h y l e n e g l y c o l(,b ) g l y c e r o l o r o t h e r s o l v e n t s .
Dilution 0.002t; in ethvl alcohol.
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NOTES
699
0 . 0 0 2 ii n e t h y l a l c o h o l , / e t h y l
c
o
c
I
ra
d a v eI e n o t h( r y )
F t c . 2 E l . S p e c t r a la n a l y s i so f c o n j u g a t e dc o d l i v e r o i l f a t t y a c i d s s h o w s t h e p r e s e n c e
o f c o n j u g a t e dd i - , t r i - , t e t r a - ,p e n t a - a n d h e x a e n e s .
700
xESEARcH rN pHysropArHoLocy
The relationship
of the differentconjugated
membersto the quenching
effecthasbeenstudied.Fatty acidmixtureshavingdiflerentproportionsof
isomerswith 2, 3, 4, 5 and 6 conjugated
doublebondswere obtainedby
conjugation
or by treatmentof conjugated
mixtureswith heat, oxygen,
0uenchino
effect
o
u
c
O
IJ
vl
a,
L
o
t
l!
bt
C o n c e n t r a t i oonf t h e C o n . i u o a t F
atty Acids
ed
F I c . 2 8 2 . Q u e n c h i n go f f l u o r e s c e n c e
o f a m e t h y l c h o l a n t h r e n e( . 0 0 6 2 9 b) s o l u r i o n i n
alcohol by different concentrationsof conjugated fish oil fatty acids.
chlorine or sulfur. Changesin the proportionsof the di-, tri-, tetra-, pentawere followedby meansof spectralanalyses.
and hexaenes
The changesin
the heightof the peaksin thesecurvescorresponding
to the differentconjugated polyeneswere then comparedwith the changesin the quenching
fatty acid mixtures.Figure 283 showsthe speceffectof the corresponding
N o r Es
701
:
6
2
6
e
F
ta
lave
Length
Crt't')
F t c . 2 8 3 . C h a n g e si n t h e a b s o r p t i o ns p e c t r ao f a m i x t u r e o f c o n j u g a t e df i s h o i l f a t t y
a c i d s i n d u c e d b y t r e a t m e n t w i t h o x y g e n . T h e t r e a t m e n th a s a g r e a t e r e f f e c t o n t h e
h i g h e r u n s a t u r a t e dm e m b e r s ,w i t h t h e p r o p o r t i o n o f t r i - , t e t r a - , p e n t a - ,a n d h e x a e n e s
d e c r e a s i n ga s t r e a t m e n tc o n t i n u e s a
. s s e e nb y t h e r e d u c t i o ni n t h e h e i g h t o r e v e n d i s a p p e a r a n c eo f t h e p e a k s .A f t e r l 2 h o u r s o f t r e a t m e n t ,t h e c o n j u g a t e dd i e n e sa r e t h e
only ones not yet influenced.
D i l u t i o n0 . 0 0 2 9 i i n e t h v l a l c o h o l .
t h e m i x t u r e s ,I n t h i s e x p e r i m e n ti t a p p e a r st h a t t h e q u e n c h i n ge f f e c t
could also be relatedto the presenceof conjugatedpentaenes.Evidence
availablefrom othcr cxpcrimentsdo not, howcvcr,suffhciently
supportthis.
We studiedin a similar way the effectinducedby the treatment-with
sulfuricacid-of a mixture of conjugatedfatty acidsof cod liver oil. Fig.
285 shows part of the occurringchangesand Fig. 286, the quenching
effect.
Similarly,wc studiedthc changesin the quenchingeffect during the
conjugationwith KOH of cod liver oil fatty acidsin ethyl alcohol.Fig,287
702
RESEARCH IN
PHYSIOPATHOLOGY
R e s i d u aF
l luorescence
'-"'
----- ---
total mixture
234dienes
2 7 0t r i e n e s
3I4 tetraenes
c46pentaenes
37iihexaenes
c
o
E
c
f!
L
;a
n 0u r s
F r c . 2 8 4 . T h e r e l a t i o n s h i pb e l w e e nc h a n g e so f t h e d i - , t r i - , t e t r a - ,p e n t a -a n d h e x a e n i c
peaks as found on spectral analysis of samples of conjugated fish oil fatty acids
treated for different lengths of time with oxygen and the quenching effect of tbe
same samples.A close parallelism exists between the decreascin the proportion of
t e t r a - a n d p e n t a e n i cp e a k sa n d t h e q u e n c h i n ga c t i v i t y o f t h e m i x t u r e .
NOTES
703
f a v e L e n g t h( 1 )
F l c . 2 8 5 . T h e c h a n g e si n t h e s p e c t r a la n a l y s i so f a m i x t u r e o f c o n j u g a t e dc o d l i v e r
o i l f a t t y a c i d s , i n d u c e d b y t h e t r e a t m e n tw i t h s u l f u r i c a c i d . T h e t r e a t m e n t l e a d s t o
u n e q u a l d e c r e a s ei n t h e a m o u n t o f d i f f e r e n t c o n j u g a t e dm e m b e r s .O n l y t w o c u r v e s
a r e s h o w n : a t t h e b e g i n n i n go f t h e t r e a t m e n ta n d a t 2 6 0 m i n u t e s .
Conjugated Fatty Acids and Induced Carcinogenesis
We have investigatedthe influenccexertedby the fatty acids---<onjugatedor not, and their mixture,upon the inductionof tumors by carcinogens.From the variousexperiments,
somewere eliminated,either because
the dose of methylcholanthreneemployed did not produce tumors in a
sufficientnumber in control animalsto permit any conclusivecomparison,
or the deathrate from intercurrentcauseswas abnorrnallyhigh so that the
entire experimentshad to be discarded.
The experimentsthat were satisfactorilycompleted are summarized
'104
nESEARcHrN pHysropATHoLocy
g
o
o
o
i\ ..
trlene
Li
c(rJ
r ng
Cuench
ie
c
c
.D
f
cl
t
f
.o
l ' ' in u te s
F r o . 2 8 6 . T h e c h a n g e si n t h e q u e n c h i n go f t h e t r e a t e d m i x t u r e p a r a l l e l t b e c h a n g e s
ioduccd in the amounts of conjugatedtetra-, penta- and bexaeoes.
NorEs
705
/1,
ir[
l^
, i
/ \'.^,
f\
,i..'\.
\.
,,
lr^tr,,/
t
t
r .
i r
,,
,r',/
\
i
"-'...j'
ll
,i ti
tr '\.
,',
".,'
'
\ \
\ \ "(a)
\
\r(b)
t
I
I
I
I
I
I
'\f
t
'r,'
l'-'
,!
have
Lenolh (np1
F t c . 2 8 7 . S p e c t r a la n a l y s e so f f a t t y a c i d s o f c o d l i v e r o i l , t r e a t e dw i t h K O H i n c t h y l
a l c o h o l .T h e y s h o w t h e a p p e a r a n c eo f h i g h a m o u n t so f t c t r a - , p c n t a - a n d h e x a e n e s .
Died
Without Tumors
Tumors
l0
3
t2/3O
t8/37
40
48
7/34
20
3,/26
II
Cottonseed oil-controls
Fatty acids from cod liver oil
Conjugatedfatty acids from
cod liver oil
Conjugatedfatty acids from
cod liver oil ( + )
1 A
%
With Tumors
706
RESEARCH IN
PHYSIOPATHOLOGY
\\.\
\
q,
(J
c
qJ
a
4o
L
o
t!
5'o
q,
-ol
o,
E
}{
C o n c e n t r a t roof nt n e f a t t y a c r ds o l s .
F r c . 2 8 8 . C h a n g e si n t h e t o t a l q u e n c h i n gc a p a c i t yo f s a m p l e so f c o d l i v e r o i l d u r i n g
i s o m e r i z a t i o nw i t h K O H i n e t h y l a l c o b o l . W h i l e t h e q u e n c h i n ge f f e c t i s r e d u c e d even for high concentration-for the sample having only 15 hour of conjugation, it is
high for that obtainedafter 24 hours. It remainsalmost thc same for the samplc
after E4 hours of conjugation. The quenching appears related to the presence of
c o n j u g a t e di s o m e r s ,w i t h 4 o r m o r e d o u b l e b o n d s .
:,
:,
I.
.;,
:t,tti
NOTES
C
C'
v,
o
707
&
E
6
c
.rt
L
F
la
.12
,62
.ott
,.b-
.@t
.@
,<b.t
!{)ol'
Concentrat
ion
Flc. 289. Quenching effect of parinaric acid upon the ffuorescence of methylcholanthrene. The relationship between tbe quenching eftect and the prescnce of
conjugatedletraenesis seen in the fact that parinaric acid has a quenching eftect of
96.2 for a dilution of 0.O06Voand still one of 62% for a dilution of. O.AOO2%.
708
nESEARcHtN pHysropATHoLocy
methylcholanthrene
injection.Thirty-threeanimals died without tumors
during the courseof the experimental
period,The number of animalssurviving for five months plus the number in which tumors developedduring
this period of time are listed for each group.
In a secondgroup of experiments(Tnnle XXXIX) six groupsof 40
mice each receivedone subcutaneousinjection of 0.25 mgm. of methylcholanthreneof an 0.ZVosolutionin tricaprylin and a secondsimilar injection one week later. Groups were treated twice weekly for three months
with 0.3 cc. of 57o solutionsof the following fatty acids in cottonseedoil:
Fatty acids from cod liver oil, conjugatedfatty acids from cod liver oil,
eleostearicacid, linoleic acid and conjugatedlinoleic acid. A control goup
of animalsreceived0.3 cc. of cottonseedoil in the contralateralflank twice
weeklv for three months.
T,rsLr XXXIX
Died
Without Tumors
Treatment
Cottonseedoil--<ontrols
Fatty acids from cod liver oil
Conjugatedfatty acidsfrom cod
liver oil
Eleostearicacid
Linoleic acid
Conjugatedlinoleic acid
-/o
Tumors
With Tunrors
4
3
3t.36
33i37
86
89
6
l0
o
6
1 5 ,3t 4
24,30
29//40
25,,34
44
80
72
73
Methylcholanthrene
f
cottonseedoil
Methylcholanthrene-lfatty acidsfrom
fish oil
r
Methylcholanthrene
conjugatedfatty acids
from fish liver oil
MethylcholanthreneL
eleostearicand
conjugatedlinoleic
acid
Tumors
% With
Tunrors
95
33i44
85
20./46
43
t9
6i46
l3
24,/39
6l
92
ll
NorE s
709
Substance
Glycerol
n-Butanol
Butyl mercaptan
Hexyl mercaptan
Dodecyl mercaptan
Hexadecyl nrercaptan
Na thiosulfate
Ethyl sulfate
N itrogen mustard
Allyl K xanthate
Nitromethane
Ethylene trithiocarbanrate
Cholesterol
Alconol_
% Dilution Used
5.0
4.5
1.0
2.0
2.0
2.0
.0-5cc. fronr
50% solution
1.0
0.1
1.0
1.0
1.0
1.0
Fluorescence
106.8
96.4
ro2.5
92.O
82.0
70.0
97.0
95.0
79.9
3.8
7.4
,7
93.0
lllElFFl-ffi*iEgiE-
710
xESEARcHIN PHYSIopATHoLocY
7ll
Control
# t
Start treataent
rith
Treated
o(- 0H
caprylicaci
tjto. 290. The administraiion of alpha oH c:aprylic acid in micc bcnring 6c.HED
lympbosarcoma tumor, induccs the dirappcaranccof the tumor in a high proportion
of cases.
or four days,when the tumor was alreadyhighly reduced,the administration of the preparertionwas discontinucd.the tumor began to grow again,
but much more slowly than usual.
Chapter 12, Note 7. Hydropersulftdes
The existenceof different bonds betweenunsaturatedfatty acids and
oxygenled us to study difTcrentbondssinrilarlyoccurringbctwecnthe same
fatty acidsand sulfur, the secondmenrberof the oxygen series.The treatment of polyunsaturatcdfatty acids or their triglycerideswith sulfur has
shown that two different formationscan bc obtained,By hcating the mixtures above ll0' but below 125'C. precipitatedsulfur is incorporated
without a manifestchangcin color or other properties.The iodine numbcr
is not changed.When conjugatcdfatty acids or thcir triglyceridesare
treated, no changesare seen in thc spectral analysis.By heating abovc
130uC, the color of thc prcparationchangcdprogrcssively
reachingdeep
prolonged.
rcd-brown if the trcatment is suflicicntly
Concomitantly, the
iodine number decreascsprogressivelyand cventually reacheszero. The
spectralanalysisof the conjugatedfatty acids shows the peaks going progressivelydown until no more conjugatedformationsare present,indicating
that thesechangesaffect the double bond.
The analogy between the fixation effectsof oxygcn and sulfur has sug-
712
REsEARcH rN pHysropATHoLocy
F I c . 2 9 l . Q u e n c h i n go f m e t h y l c h o l a n t h r e n e
f l u o r e s c e n c ei n s a m p l e so b t a i n e d w h e n
0 . 5 % s u l f u r i n c o t t o n s e e do i l w a s h e a t e d f r o m 1 2 0 " C t o 2 8 0 ' C . T h e s a m p l e sw e r e
d i s s o l v e di n e t h e r - a l c o b o lm i x t u r e a n d m i x e d i n e q u a l p a r t w i t h . O l 2 S % m e t h y l cholanthrene.The existenceof concomitant changes in the oil with sulfur and in
the oil alone, indicates that the variations correspond primarily to cbangcs which
t a k e p l a c ei n t h e o i l i t s e l f w h e n h e a t e d .
NorEs
713
714 I
R E s r , A R c HI N p H y s r o p A T H o L o c y
Glycerol
Eefone
After
Ftc. 292. Electrocardiogram of a rahbit receiving intravcinous injections of a rolulion of glycerol ?0f1. characterizedby the appearanceof extrasystoles.(a) bcfore
treatment. (b) after 30 cc.
Butarrcl
F 1 o . 2 9 3 . E . r t r a s y s t o l eas p p c a r a f t e r t h c i n t r a p c r i t o n c a li n j e c t i o n o f b u t $ n o l i n v e r y
h i g h d o s e s .( a ) b e f o r e t r e a t m e n t .( b ) a f t e r 1 . 6 g r a n r / 1 0 0 0g r o f a n i m s l .
NorEs
715
n E s E A R c Hr N p H y s r o p A T H o L o c y
716 /
C
o
i n e t h v l a l c o h o l / e t h yal l c o h o l
Lyrphnodes
Adrenalgland
Sonetumrs
t y p e0
Io
L
La
f a v e L e n o t h( m l ' )
NorEs
717
t J . 4 Xs o l , r rt o n
c
o
E
6
c
4'
t-
}{
l'
J@
fraveLenoth(m r.)
Frc. 295. Details of the spectral analysis of rat colostrum indicates the existenceof
a formation with three peaks in the region 290-250 mp with some similarity to tbe
c o n j u g a t e dt r i e D e s .
718
xESEARcH tN pHysropATHoLocy
NorEs
'l19
necrosiswhen administeredintramuscularly
in animalsand to inducepain
at the site of injectionh humans.A 307o solutionof butanol,however,
was well tolerated.Daily administrationof subnarcoticdosesfor long periods to micecausedno toxic effects.On the other hand, repeated injections
with narcotizingdoseswere toxic and even led to death of the animals
after severaldays.
Chapter 13, Note 9. Butanol and Leucocytes
T h e a d m i n i s t r a t i oonf b u t a n o li n s o l u t i o n so f 6 . 5 % i n s a l i n ei n t r a p e r itoneallyin rats was seento inducea hyperleucocytosis.
5 cc. injectedat
once was seento double the prcviousamountof leucocytes.
This increase
6l
o
>
o
I
J
o
J2
6() :to
Jo
oo
so
lLo
,r"
tuo )ra
rar
rj,
160 te
tzo ,60
i,rr?rtff
F t c . 2 9 6 . T h e a d m i n i s t r a t i o no f 5 c c . o f a s o l u t i o no f 6 . 5 % n - B u t a n o l i n t r a - p e r i t o n e c l l y t o r a t s i n d u c e sa n i n c r e a s ei n t h e l e u c o c y t en u m b e r .
s t a r t e dt w o h o u r s a f t e r t h e b e g i n n i n go f t h e i n j e c t i o n sa n d c o n t i n u e d
prog5cssively,
to reach the valuc of 34,000, seven hours after the bcginning of the injections.The hyperleucocytosis
was seen to persistfor
hours.
was
more than 24
The numberof leucocytes increasedin the animal
s h o w ni n F i g . 2 9 6 .
A s t i l l m o r e m a n i f e set f f e c tw a so b t a i n e dw i t h i n j e c t i o n o
sf I cc.of the
e v e r yh o u r d u r i n gt h e d a y . I t i s i n t e r e s t i ntgo n o t e
s a m es o l u t i o nr, e p e a t e d
that this effect was manifestedalmost 6 hours after thc injection with
b u t a n o lF
. i g . 2 9 7 s h o w sa n e x a m p l eo f t h e s ee x p e r i m e n ti sn w h i c ht h e n u m arrivedat 42,500.
ber of leucocytes
'720
nesEARcH rN pHystopATHoLocy
4{w
o
>
(,
CI
.J
J
3 4,no
60
Jo
Jo
90
/2o
/t0
lto
2to
2(o
27o
J@
JJO
J6O
J9O
.aO
Hinutes
Ftc. 297. The administration of I cc. of n-Butanol, repeated every hour, induces a
sizable increaseof leucocvtesin rats.
;ir:it:!
NorEs
721
'122
nEsEARcH rN pHysropATHoLocy
In February 1944, the pain reappeared.Study of the pain curve revealedit to be of the acid type sincethe paroxysmalpain correspondsto a
very low pH. The patient was given ammonium acetate and lipoesters.
Four days later the pain had considerablydecreased.and fifteendays later,
completelysubsided."
"Mrs. W. For elevenyearsthis patienthas sufferedof a left trigeminal
neuralgia.Each year the crisislastedfour to five weeks,during which time
the pain always appearedbetweenfour and seven in the morning, lasted
for one to two hours and then disappeared.
The pain was so severe,being
almost unbearable.For the rest of the day, the patient only felt a slight
sensitivity.Barbituratestaken even in large amountshad no influenceon
the pain. Removal of the Gassergangliawas suggestedas the sole possible
cure by numerousdoctorsconsultedin Mexico and the United States.
The patient came under our care on November 12, 1943. Her most
recentcrisishad startedon October3l . Studyof hcr pain showedit to be
of an alkalinecharacter,sincepain is quite intenscwhen the urinary pH is
glycerine.The patientprotested,feelingthat a few
high. we recommended
dropsof glycerinewould not be ableto help her, pointingout that intensive
treatment,had given no results.However,on November l5th, when pain
startedat 5:00 A.M.,the patienttook the glycerinedropsand to her amazement, the pain disappearedwithin two minutes.She assertedthat no medicine had ever beenable to stop the pain once it had started.At 7:30,r.rrr.
the pain returned but again decreasedafter the patient had taken several
glycerinedrops.
O n N o v e m b c rl 6 t h , t h e p a t i e n te x p e r i e n c epda i n a t l : 3 0 , 5 : 3 0 a n d
7:30 n.t"r.During the first two periodsof pain, the pain was instantaneously calmedwith glycerineand phosphoricacid; the third period of pain
was decreased
in intensitybut lastedfor 40 minutes.
on NovemberlTth: pain appearedat 9:00 l.r',r.but disappeared
three
minutesafter medicationof glycerineand phosphoricacid.
O n N o v e m b e rl 8 t h : p a i n w h i c h s t a r t e da t 8 : 0 0 L v . c o u l d n o t b e
calmedwith medication.Coramine,Cholesterol
and insaponifiable
fractions
rverethen prescribcd.
o n N o v e m b e rl 9 t h : p a i n w a s e x p e r i e n c eadt 4 : 0 0 a n d 8 : 0 0 p . r r r .b u t
subsidedwithin three minutes after medicationof glycerine,phosphoric
acid and coraminewas taken.
on Novcmber20th: from this day throughNovember25th, the medication was unableto influenccthe pain, and consequently
the patient becamedisheartened.
A study of her pain patternat this point indicatedthar
it had changedto the oppositetype-from alkaline to a defrniteacid pattern.
on November 26th: bicarbonatewas given at the onset of pain resulting in a considerabledecrcasein its severity.No further pain was felt on
the following days and the crisis was consideredended.In this instancc.
the crisishad lastedfor 27 days (the lengthusuallyvaried from 27 to 35
ror"l,
days). However, the treatmentachievedwhat had been;;::rr.',
previouslytried treatments-the cessationof pain once it had started.
These observationsled to the following pertinentconclusions:1 ) they
showedthe existenceof typical acid or alkaline pain;2) the possibilityof
changingpain from one type to the oppositeone, either during the course
of the diseaseor due to medication;and 3) the possibilityof eliminating
pain with appropriatetreatment."
Chapter 14, Note 2. Dr. Welt's Publicationon Butanol-Conclusions
"n-Butanolwas administered
to a largenumberof patientswith pain duc
to the trauma of variouscommon otorhinolaryngological
and ophthalmologic surgicalprocedures.Pain was relievedin approximately90 per cent
of the patientsso treated.
Theseclinical resultswere consideredin the light of studiesby Revici
and his co-workersregardingthe physiopathologyof wounds. The results
indicatethat the proposedconceptof pain has significantpracticalclinical
applications."
Chapter 14, Note 3. Dr. A. Ravich'sConclusions(189)
In his articleconcerningthe post-opcrative
A.
care in prostatectomies,
Ravich arrivesat the followingconclusions
298.
exemplifiedin Figure
SUMMARY
" A n e w c o n c c p to f t h e l o c a l p h y s i c o c h c m i ccahl a n g e so c c u r r i n gw i t h i n
pathologicalfoci as introducedby Revici, has beenbriefly described.Accordingto this view, pain is the resultof local pH changesbrought about
by the accumulation
within disturbedtissues.
of acid or alkalinesubstances
Changesin the lipid balanceare associated
with and may accountfor these
alterations.
"Thc possibilityof correctingor neutralizing
suchlipid changeshasbccn
exploredclinically in severalseriesof urologicalcases.Thc favorableeffects
upon pain as well as upon bleeding,wound healingand other important
postoperativeproblems and complicationsindicate the need for further
study along theselines."
Chapter 14, Note 4. Treatment of Post-traumaticConditions
Of spccialtheoreticaland practicalintercsthas becn the treatmentof
traumaticconditions,especiallythose following surgicalprocedures.The
recognitionof the role of fatty acids acting at differentlevelsof the organizationand inducing severaldifferent manifestations,
has led to the
concomitantuse of variousagentsproper to the levels.From the various
agentsstudied,heptanolwas thus chosenas acting at the cellular level,
glycerol,polyunsaturated
alcoholsand alkaline amino acids and butanol
724 /
RESEARCH IN
I)ayof
PHYSIOPATHOLOGY
NorEs
725
726
xESEARcH rN pHysropATHoLocy
Reliefof Symptoms
NumMod.a r K ber
Slight erate
ed
z
l
44
2
l
I
5
2
0
J
4
2
0
2
4
0
l
3
J
0
I
2
3
0
( 6 0 %)
l
0
0
l
1
l
0
0
0
0
I
0
I
I
Temporary
l Aresl'
I
I
s -
Regression
2
0
i ') l
I
i
i
6
0
0
I
0
0
0
I
1
2
6
(24%) (t2%)
T,rsr-pXVIB
Resurrsor LtproTHenrpyAloxr,
TemTotal
Type of
NumMalignancy
ber
Breast
t4
Lymphoblastoma 8
Lung
3
Head and Neck
3
Gastro-intestinal 6
Gynecological
)
t
Sarcoma
I
Miscellaneous
2
Total
porary
Reliefof Symptoms
NumMod- MarkArber
Slight erate
rest
ed
-2
1
0
6
5
0
0
5
3
3
0
0
3
0
2
l
0
l
0
3
l
l
l
0
I
0
0
r
I
0
0
0
0
0
2
0
0
2
I
23
l9
7
(58c/c)
Regression
2
2
0
0
I
0
I
6
(t8vo)(ts%)
BIBLIOCRAPHY
728
RESEARcH lN
PHYSToPATHoLocY
1947.
25. LE FEVRE, R. J. W.: Dipole Moments. New York. Chem. Publ. Co..
1938.
26. PENNY, W. G.: The Quantunt T'heory ol Valency. London. Methuen.
1935.
SIDGWICK, N. V.: The Elecnonic Theory ol Valency.Oxford. Clarendon
Press, 1929.
GORTNER, R. A.: Outlinesol Biochemrstry.London. Wiley & Son. 1947.
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r949.
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305. GARDNER, W. V.: Studieson steroidhormonesin experinrentalcancer.
R e c .P r o g r . H o r m o n e R e s . l : 2 1 7 , 1 9 4 7 .
306. GREENSTEIN, J. P.: Bictchemistry
<tl Cancer.New York, 1954.
307. HAUSCHKA, T. S,: Inrmunologicaspectsof cancer: a review. Cancer
R e s .l 2 : 6 1 5 , 1 9 5 2 .
308. KALISS, N., and MOLOMUT, N.: Tbe eftectof prior injectionsof tissue
a n t i s e r u m sC. a n c e rR e s . l 2 : l l O , 1 9 5 2 .
309. LEVAN, A.: Chromosomesin cancer tissue.Ann. N. Y. Acad. Sci.
63:774, 1956.
310. LURIA, S. E.: Mechanism of bacteriophagereproduction.Fed. Proc.
1 0 : 5 8 2 ,1 9 5 1 .
746 /
R E S E A R c Hr N P H Y s l o P A r H o L o c Y
IIiDEX
A s N o R v ^ , r .a m i n o a c i d s .. 1I , l ? 0 . t - 5 - l
- i ; r i l r ; r c i c l sl.i l l
,rnrl !r)fllur:.ill!lfr. I l(].
fr"(i
i I l.
: is.
fr,tr
I i-i
il'l
llllrlil].rr,,l,rr'.
. l t l r j l . r t i r . r i t' n ' i i l 1 1 q ' . r . I l t i
, i t ] \ lr . , i , . r . l l t \ j - . S i I r
-,
. - ; r 1 1 r sJ l l , ' . 1 . . . ' . 1 l ] ' . } . l i ' r
'
"
\.'r',,
. r ' l i i r l ; t ! : : t r , r \ r i t , t ' i ( l[ ) r ) l i r \ r i r i n t .
lr..i{}irilt lti..t,fl,.:trtJr,rirltttti. "111
\ , i , r ; . . , r , , , 1 . .] j
\ , j r : t , , r r t , r ' , r i r r. l l , r r , r . l r . t , ! l l L i l | t l r : . ? l t i
l ' ' , , . l . r l r , r l l i . i, / i l t \ . i { l l
\,1ti.,,|. ii'llr.rrlr1 1r|11j. ltrt}
, tl,j ,l;llIt
fiti:\l.ritl. j-lfr, .l"ll. 6{i
.\,ltt 1liirclrr1l\
.rtrrl ,il.1r]t tlt;ll fitlll, irCir.lt,
.l(l
! l t ) \ t ' l l l t l l r ' i r l : ! ! ' f.
.\r'r'l\lrlLlr.rlir,rii,lI'il-l
. ' \ i i , l 1 . , ' r r l ' ; r l , r r i , r ' i l r , , ' , 1 .i { (
. i ' ' i t l l ! i t ' r r: I i , r i r r ' . { i l . . . i - - - { '
'',r
l l l ( , \ \ n r l c , r .r , l .
-'.
i n i n t n : r i r r : ,111 . : . , t r n t: .t l.
i,l
(
,
l
'
:
.
.
1
i
l l l i l i h i n * , 1 ,1
" i n i t l l t t t i r( i f l l l L ' r \ i \ t ' i . , l r ' r . l , f
irrp:rirt .lt, i.'1. ii-\. ir I
i.',,
: , ; 1 : . 1 l , - ' i l L r l , - . t ; r l i , . t iti.\ : . I , i f i
, l r l , . l 1 , ,r t t l . i l . l l t . ' , . \ i ! . I n . l l ) i ,
i t t . - ll l t . i r , 1 r ' ! 1 i 1 1 1 it<. i
i(tri
, r i r , il r , , . i r i , . l , r : , .
l-'\
lll \r'lii|',. f)'1!
ll1 '.\\llil1rl\.
. \ c t r l . i r , r r cl l \ \ l i r 1 i : f t r r j r l r t , \ . \ . .
'\ri,llfrrttS;rg,.'r:- i*
Al
r r ( 1 ." i
. . \ e i t l l r r t l t r r ] ' .1 i '
Aciti
-
lr.lrll,,tt.
irtr
.lil
\ri
l')l
\1irrrrt,l1
. r t ) l t i . l r . n . ,I \ { - }
r r l r t r l r t r l i r r , r ' 1l 1l r r ; 1 1 1 l. r l r )
i(rt
\l.ttr1'..'nlllj(ln{inir,igiC,.1"ll
, r u ( i [ r . i i r .: . i ] :
- - * ; r r t r iS . L i t - .
;t[1. t'li]
..
.- 61 t1l.fl!.lL'l,i,rill:. l{)1.:{}i
":t}r
- - l I : l l l r l ' i t ) l l l . r l " l \ f l . r l l i ! , .f r q i . - I t '
-'r
.\uttl trnri .'li,.,lrri, i:,ri:arJl.,'i iii.,l:nt,'.
' 1 1i
l l l l l ' 1 1 r1: t r ' , i i r r , l l i l l ! t . l i -
t . r r i i , , i i i ' t rl '. -
\ i l r . r ; r r ' i i i s l t r ( . r ' . r 1 , 1r l r t r r k . . l : i l
lipirlri l"t.rttit,ttr i II
lll fllflel': lllil,il.!.
I i'r
, i l t r l i i t ' l t . t r r U .l r . l { l
-l-'
: r r r r li l . r f l t . . ' \ , i r l 't . , 1 t .
.
l
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. . \ t ' r ' i i r ; r r r , l l. l l
j
5t lf :ttit1il. i:l
Itl Lrrirlf. rrl]
li.)fr
\ . , . r r l l t . r \ i : l i \ .l , ' , . r t r , l i , r l t i r : 1 .] l \
\,ritr, 'ltLrrf. ,t.t \1r,,.1.
t l C i r i l l l r i l l 1 r l g , 1 f r . 1 l\ \ i ( l r , i i t l
-1,,
Il-'.
- , \ l \ n { , i l r l . r l i t i r l i , , r r . { , ( . 1 I. i i )
.
i r t , l i r 1 r 1 , ; 1 r j 1 1 1 ;I,' .i l
\\.fla,\.li]/|
\li;nlljti
I l 1 e 1 1 1 , ) ( il 1
l'l;trl'il.rit,ti1.
/,,'i,. {i-'-'
frll.
"
A c r o l ei n . 3 3 0
, { C ' l l { l r r r , .l , l l , r i - .:rrrl . , r ).f f r l i
I t t r l r l r r l r r tt i i ' ^ ; . l l
-. .r.l
cfr i
ttl ll;|il. .:l
i
llt llnrt,r'.. l{}1
-{},
."si*tlai(int!,
i{,. f.l.
.llil
lIr lr:rli!:rl. r,(). 'i:S
i:,.i1i,trit:rgrp.rr{i1...l'i(l
t r ! t l t r . l ; ! t ) \ . . . l i f . r{ .i i
t r g g l l i r r ! 1 , l i , r r t l. : l ) .
iri.l
l(ii}. |}-l
t,i('
748 /
rxoex
Analytical dualism,86
- patterns,90
A n a p h y l a t i cs h o c k ,l 8 l
A n d r o s t e r o n e l,4 I
A n e s t h e t i c si ,n d e x o f r e p a r t i t i o n ,1 2 5
A n g i n a p a i n a n d p r o c a i n e ,3 9 2
A n i m a l s , p l a c c i n o r g a n i z a t i o n ,3 0
A n i o n s i n S . d . c .p H , 5 9 9
A n o x y b i o s i s ,9 5
Antagonismagents,seepharmacodynamic
activity
- b e t w e e ne l e m e n t s ,1 0 4 - 1 0 7
- - falty acids-antifatty acids, I4g-
r65,362
INDEX
Anuria..ll7
Arrchitlunic acid and rdrenals.316. 373
* - - * a n d u l l o p r e g n a n e6, 3 |
- - - - - a n c lb l o t i l r e t l c e l l r . 3 1 6
* - and corticoid\, | "19
* - a \ c r s e n t i a lf a t t y a c i d , 6 ? I
--- - irnd qucnching tif llut>resccncc,
697
- * - * a n d S . d . c .p H . t r Ol
- - ' . a n d r m a l l p o xv i r u s .I l 6
- * a n d r l e r o l s ,I 1 6 . 6 l t
,\rginine, t 69
- l n d e o r i n o p h i l e r .- s 6 9
' i n r ' r ; l r r n i r ; i l i r '1r S
r.
. \ r n c l l r . l u i - r t t r r l l irl n i i I r [ . r \ l \ ,( r t ) S
,.\r.tnir: ,rnrl lrirc. ii,rlijr,tiil. J{l:
-,r\
r a r f ! ' i l l r r g u t 1t .{ t t )
- - - - a r r L i t t - ft l ] { r g f t } . J l } , i
, \ f [ ( ' f] { ) \ !i r ' t i r r i s .J \ l
.
. r t i t l , l l . i l l l , r l r . . li r
'lllt('f\'ill.i, .i{ I
.
.
.rlltl liil,rflti',rrlrrll
B c r r r ' l l i u n r ,J - 1 4
[ 3 e t r g k r h u l i n s ,l l l l
B e t l n r c r e i r p t o p r o p a n ot ci ci d . , 1 3 9
B e t ao r i d a t i o n .l : t t . 1 3 1 .6 1 6 . 6 l l t
- - * * . i n r a n c i d i { y ,l l t {
B et l n r d i a t i o n .l . l I
[ ] i c i i r h o n a t ci o n i i n d S . d . c .p H . 6 0 0
1 3 i o l o g i c af al c l o r s a n d S . d . c .p H . 6 0 8
- p f r l p c r l i c Joi f l i p i d s . 1 2 . 5
- r e i l l m .l 7
- -- houndirryfornlalion. 23
B i o l o g i c a l l yg u i d e ' dc h c m o l h e r a p ) "5. 2 6
" .rr' ? 6 1
r ; , . i i' ,l l r r ' r p
t l l r r ' r p ' ., ' l I ' l
l l j r r i l r r l i rI ,{ } 5 ,- 1 { ) 1(.} 1 ) ( i
l l i r r i l l l r lr r c g i r t i v. tr ': l e
r r i r r r l3
, Jl
' u l f r r r l. J ( l
l l i ' , i r r t l' i ) I
, r l i ' 1 r : i 1 1 1 1 9 L 1 l r .cr " ' l l r . t l J
, 1 1 . 1 , . , ' l l ! l r i . i \ r l t i.^ ' . 1
1,1"1
ii
i::rilrlrl..
a
.1
"-
.\ltiriitr'. J.i5
.,\'tili.r
.,rtl r l
llllri,l
it!r,!ii\
, r l l l l l \ r i r - t r t t1
, r l r . i l .i l r
t | t c i t i l t , r i t i i J t t : ti . l t
'
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llIl
l|r\llll(
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ill.t,ll
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.l.l'l
ll.
ltr
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ic i
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. i l.'. {rfrS
.rlirt rrrtilLrtl).i't;
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'
'
'. '.
(rlr
rll
lllrr'l{:l-lr)nllirut'\. .('r
llli.iclil.l,rllr
i{r.i
iirl
l,trl
ittllttr.ir..'. f,' {
ll,rr,,irli1l1ti11ip,rr;ti,''iiir
'
l J 1 1 ' 1 r r ] 1 1 i tr i. i i ' r l l , - ; 1 , r
l l r r t l . r t t l i l l . ! L L l l r , i 1 r . i t , , i .1 1 , . , i l l l lrrr: l l l . ; r r t t r
tt]11t1"
s"I
l l i , , r i t r r l r l t r r i l i t r r r' , f i i l - . i t l . , I l - l
l { , ' , 1 , ,. 1 ; 1 , 1l r l r r t i r ; r l l r l t r . r -: .i t ,
. ; r t t l l t t i r r r : . i l t r l . : i ! r ' 1 1 : , - ^. i i l l
r'a!(l
iirl\l llllirj.' l '-.
lril,ilcs
r11r'rl rrl il'rr'r{\1.
ii-".
l l ( ! 1 , t \ . l r i | l r ,. 1 t i , 5 { , .
. , \ l r r i \ J ) i l i ! ' \ 1 , 1 l. \ r r
I t . j , , ' 1 1 r .\ ( ' r | ( ' i l ! ' l ] \
.rr'.1
l r : r r 'r r ' , 1 r l l . . : t ' ' . ' i
{ , 1 ,I
,rllrl linjtrjr.
llrt
ii,.i
frr'llr
-iirl
l i ' , t . , i : . l ri ' t
I i . ' r t . jt ' r 1 ' . r ' l r. r , r , . { ...f r i r , r i r l r , ' Iq,.I l . J { 1 . 1
t , ' r
li-
750
rNDEX
Caffeine, 604
C a l c i f e r o la n d c a l c i u m , 3 5 8
Calcium in blood, 86
- and calciferol,358
- andcancer3
, 5 6 , 3 6 1 ,3 9 5
- a n d c a r c i n o g e n s3, 5 7
- a n d c c l l u l a rm e m b r a n e .1 2 6
- c h l o r i d e ,4 0 3
- and copper,359
- a n d d u a l i s m ,E 6 , 1 0 2
- e l e m e n t s ,3 5 6
- excretionof, I 66. 595
- a n d f a t t y a c i d s ,1 2 6 , 3 5 8
- and grass tetany, 355
-index, 575, 595
- i n d u c e dc h a n g e s 3
, 57
- a n d l i v e r c e l l r e g e n e r a t i o n3, 5 6
- a n d p a t t e r n s8, 5
- a n d p h o s p h o r u s ,3 5 9
- r e t e n t i o n ,1 6 6 , 3 5 9 , 5 7 5 . 5 9 5
- a n d S . d . c .p H , 3 5 7 , 6 0 0
- and testosterone,358
-and zinc.40l
C a l o r i c m e t a b o l i s mo f f a t t y a c i d s , 1 2 9 .
I 35.361
- - r o l e o f g l y c e r o l ,3 6 3
Cancer, sce also canccrous
- seealso tumors
- a b n o r m a lg r o w t h i n , 4 0
- allergic reaction as test in, 267
- allotropic resonanceforms, 302
- a n d c a l c i u m .3 5 7 . 3 5 9
- a n d c a r b a m i ca c i d , 3 0 7
- of the cervix and circumcision3
. 0i
- and chloridecontent, 312, 710, 716
- c i r c u l a t i n gc e l l s , 2 6 6
- and copper,l l l, 360
- d i a g n o s t i ct e s t s ,2 6 5
- and divorce, 308
- d u a l i s m i n p h y s i o p a t h o l o g yo f , 9 9
- e v a l u a t i o no f r e s u l t s ,4 1 7
- a n d e m o t i o n a l r e l a t i o n s h i p ,3 0 8 , 3 0 9
- and fatty acids. 312, 323, 326, 466.
4 7 3 , 5 0 1 ,5 3 3 , 7 0 3 , 7 t 0 , 7 t 5
- s e ea l s o f a t t y a c i d s
- and genetics3
, O 2 ,3 0 7
- a s h i e r a r c h i cc o n d i t i o n , 3 8 . 2 6 8
- immunological problems, 212-Z2O
- induction, see carcinogenesis
- i n s i r u ,3 9 , 4 0 , 3 0 5 , 5 5 0
- i n v a s i v e p h a s e ,4 0 , 3 0 5
- and lipids. see specific lipids
- and lipoids, see specific lipoids
- and magnesium3
, 55
- a n d m a r i t a l s t a t u s ,3 0 8
- a n d m e r c a p t a n ,3 3 2 . 4 7 7
- ngn-inyssive phase, 39
INDEX
Cancer as organized condition, 38
- painful phase, 40
- pathogenesis,264
- physiopathologydualism, 99
- plurality of phases, 38, 40, 264
- a n d p o t a s s i u m ,3 9 6
- preterminal phase, 40
- of prostate and circumcision, 305
- o f p r o s t a t ea n d z i n c , 4 0 l
- psychologicalfactors, 308, 3 t0
- r a d i a t i o n ,2 6 1 , 2 6 2 , 5 2 4 , 6 8 3 , 7 2 6
- a n d s m e g m a ,3 0 5
-and
sterols, 148, 158, 369-373, 466,
7t5
- and sulfur, sec specific agents
- t e r m i n a l p h a s eo f , 4 l
- therapeutic approach to, see cancer
therapy
- and unsaponifiablefractions, 306,372,
468
-- and urethane. 689
- viruses.290-301
- and widows, 308
C a n c e rt h e r a p y ,a c i d l i p i d i c f r a c t i o n ,4 6 8 .
533
-b u t a n o l .4 9 3 . 5 3 3
- - chemotherapy,463, 542
- - c o d l i v e r o i l f a t t y a c i d s ,5 3 3
- - c o n d u c t o f t r e a t m e n t ,5 3 6
--conjugated
fatty acids, 501
- - criteria used, 532
- - g l y c e r o l ,4 9 3 , 5 3 3
- - group of agents in, 473
-g u i d e dt h e r a p y , 4 6 1 , 4 6 3 , 5 2 6 , 5 4 7
- - g u i d e d l e v e l c h e m o t h e r a p y5
, 26
- - m e r c a p t a n s4
, 77
- - p l a c e n t ae x t r a c t s ,4 6 3
- - p r e s e n tf o r m , 5 3 1
- - r a d i a t i o n 2, 6 1, 5 2 4 , 6 E 3 , 7 2 6
- - radiotherapy, 524
- - r e s u l t s ,4 6 3 , 5 2 6 , 5 3 7
- - selenium5
, 1 2 ,5 1 8 , 5 3 3
- - s t e r o l s ,4 6 6
- - s u l f u r i z e do i l , 4 8 1
- - t e t r a l i n ep e r s e l e n i d e5. 1 8 , 5 3 3
persulfides, 49 l , 5 33
- - t h i o s u l f a t e s4, 8 2 , 5 3 3
- - u n s a p o n i f i a b l fer a c t i o n s ,4 6 8 , 5 3 3
C a n c e r o u sa m i n o a c i d s . 3 0 7
- c e l l s ,4 0 . 3 0 3
-chromosomes, 39. 41, 303
- e n t i t i e s a s o n t o g e n i ca l l o t r o p y , 3 0 3
- h i e r a r c h i ce n t i t i c s ,4 1 , 3 0 3
Capillary hemorrhage in shock. 228
C a p r i ca c i d ,l 2 l , 6 3 9
Caproicacid, l2l, 639
C a p r y l i ca c i d , l 2 l , 6 3 9
751
C a r b a m i ca c i d , 1 7 0 , 3 0 7
Carbon, 108,334
C a r b o n a t ei o n s i n s h o c k .2 3 0
Carbonium ion, 279
Carboxyl, resonanceforms, 8
Carcinogenesis,allotropic resonance,302
- a n d a r s e n i c ,4 0 3
- and copper, 360
- and croton oil. 690
- energeticfactors, 270
- epoxides,28 I
- e x p e r i m e n t a l2, 6 8 , 3 0 7
- and genetics,302
- and host, 295
- and lipids, 304
- and lipoids, 296
- a n d n i t r o g e n m u s t a r d ,2 7 E
- a n d s y n j u g a t ef o r m a t i o n s ,2 E 3
- t w i n f o r m a t i o n s ,2 7 1
- unsaponifiablefractions, 690
- a n d u r e t h a n e ,3 0 7
- a n d v i r u s .2 9 0 . 3 0 1
Carcinogenic hydrocarbons, fl uorescence,
695
Carcinogens,see specificsubslances
- and calcium,357
- and energetic spectrum, 288
- a n d f a t t y a c i d s ,7 0 5 , 7 0 8
- m o n s t r o s i t i e s a n d m u t a t i o n s ,2 6 8
C a r d i a cr h y t h m a n d g l y c e r o l ,3 6 6
- - a n d l i p i d s , 1 6 9 ,7 I 4
- - a n d l i p o i d s ,6 6 1 ,7 1 4
C a t a b o l i cp h a s e o f w o u n d s , l 6 l
C a t a l a s e sl ,l l , 3 6 0 , 4 0 9
Cations, see elements
- of different comparlment, 27
- a n d o r g a n i z a t i o n 2, 7
- i n S . d . c .p H , 5 9 9
C e l l s ,p l a c e i n o r g a n i z a l i o n 2
,3
- c a n c e r o u s4, 0 , 3 0 3 , 3 0 5
- - i n p r o s t a t e ,3 0 5
- rapid aging, 82
C e l l u l a r a d h e s i v e n e sasn d c a l c i u m , 3 5 7
- b a s o p h i l c y t o p l a s m ,E 2
- m e m b r a n e s1, 2 7
- o x i d a t i o n ,1 6 6
- vacuolation in shock. 228
-youth, prolonged, lO2
C e l l u l o s eg u m , 7 1 5
Cenapse, lipids, 622
Ceruloplasmin and ammonium molybdate,399
- a n d c o p p e r ,3 5 9
Cervix cancerin Jewishwomen, 305
C h a i n ,r i g i d i t y , 1 3 7
Character of tumors. adenocarcinomatous,716
752
TNDEX
270
Conductof treatmentin cancer,535
red cells, 179
Conglutinated
rNDEx
Conjugated fatty acids and ascitescclls,
't1)
753
C o n v u l s i o n sa n d h e p t o n a l , 3 8 3
- and lipid, 163,370, 659,715
- and magnesium3
i n c a n c e r t h e r a p y ,5 0 1
, 55
- and octanediol, 3E7
a n d c a r c i n o g e n e s i s7 ,0 3
- a n d s t e r o i d ,4 4 8
and carcinogens,705
- i n d u c e d b y t h i a m i n e , 3 l 3 - 31 7 , 3 2 4 ,
from cod liver oil, 335
a n d c o n v u l s i o n s3
, 24
356.44E
and fixation of chlorides, 135
Copper, 358-360
- and calcium,356
a n d l i v e r r e g e n e r a t i o n3, 2 3
- a n d c a n c e r ,I I l , 3 6 0
a n d l y m p h a t i c s y s t e m .3 2 3
- d i s t r i b u t i o n ,1 0 9 , I I I
a n d m a n i f e s t a t i o n s3, 2 2
- - - and neoglucogenic corticoids, - i n l i v e r c e l l s . I I I
- i n p e r i o d i cc h a r t , 1 0 5
362
- a n d S . d . c .p H , 6 0 0
and oxygen,701
p h a r m a c o l o g y ,3 l 9
C o r a m i n e a n d h e m o r r h a g e ,4 4 0
a n d q u e n c h i n g o f f l u o r e s c e n c e , C o r o n a r y o c c l u s i o n ,3 7 0 , 4 5 9
- t h r o m b o s i sa n d m a g n e s i u m 3
699
, 55
and S.d.c. pH,323
C o r p u s c l e s s, u b n u c l e a r ,5 4 3
a n d s h o c k ,2 2 7
C o r t i c o i d s ,1 4 4 , 3 5 5 , 3 6 7 , 3 7 3 , 3 8 3 , 6 3 5 ,
transport of, 694
637
- and aminosugars1
t r e a t m e n t so f , 7 0 1 , 7 0 2
, 45, 437, 639
-and
and tumors,322
a r a c h i d o n i ca c i d , 1 3 9 , 1 4 3 , 6 3 1
- c o m a . 1 6 3 .6 5 9
and viruses, 322
- f o r m a t i o n s ,1 3 5 , 6 2 2
- c o n v u l s i o n s , 1 6 3 .3 8 7 , 6 5 9
- i5sms15,see conjugated fatty acids
- and defense6
, 40
- linoleic acid, 335, 639, 708
- a n d e o s i n o p h i l e s5, 6 9
- pentaenics,701
C o r t i c o s t e r o n e3, 6 5
- polyenes,283
C o r t i s o n e ,1 4 5 , 4 3 5 , 4 3 8 , 6 3 6 , 6 3 7 , 6 3 8 .
- tetraenics,700
640
- t r i e n e sa n d a d r e n a ld e f e n s ei n d e x ,6 9 5 C o t t o n s e e do i l l i p o a l c o h o l s ,3 8 8
- - a n d r a d i a t i o n .2 3 8 , 2 4 E
C r i s i s i n p n e u m o n i a l. 9 E
- - and trauma, 375
C r i t e r i a f o r t r e a t m e n t ,4 1 5 , 5 3 2
- t r i e n i c a l c o h o l s ,7 1 8
Critrcaloxalic acid index,227
C o n j u g a t i o nm e t h o d s ,1 2 9 , 1 3 3 .2 3 8 . 6 7 9 , C r o t o n o i l a n d c a r c i n o g e n e s i s6,9 0
695
C r o t o n i c a l d e h y d e ,3 3 0
Connective tissue,amount of, 102
Crotyl alcohol, 389
- - a n d b i x i n e .3 2 4
Crust of the earth, constitution, 108
- - conditions, 437
C y c l i z a t i o no f a r a c h i d o n i ca c i d , 1 3 9 , 1 4 3 ,
--and
g l u c o s a m i n e1, 4 5 , 4 3 8 , 6 4 6
631
- - healing,161
- of squalene,t 39
- - and radiation, 256
Cyclopentane forces. | 39
- group and twin formations, 273
Constancyand organization,33
C o n s t a n t so f t h e e n t i t y , l 7 l
C y c l o p e n t a n o p h e n a n t h r e noer i,g i n o f , 6 3 |
- role in organization, 28
C y s t e i n e ,3 4 0
C o n s t i p a t i o n ,E 3 , 1 0 2 , 1 6 6
C y t o c h r o m eo x i d a s e ,3 9 9
- - and copper, 3-59,360
Constitucnts, 104, 222
Constitution, crust of the certh, 108
Cytologicaldualism, 82
- of viruses. 690
C y t o l y t i c a c t i v i t y ,6 9 1 - 6 9 4
C o n s t i t u t i o n a lr o l e o f f a t t y a c i d s . 1 3 0
C y t o p l a s mi n c a n c e r ,4 0 , 5 . 5 0
C o n v u l s i o n sa n d c h o l e s t e r o l3, 7 0 . 7 1 5
C y t o p l a s m i cv i r u s . 2 9 2
- and coramine, 448
- and deoxycorticosterone,367, 659
D,ux color of blood. 678
- a n d e t h y l m e r c a p t a n3, 3 2
D e a d v i r u sv a c c i n e , 2 l 9
- and fatty acids, see specificagents
Deafness,see Impaired Hearing
- and glucose, 36E
D e a t h i n c o m a , o r c o n s c i o u s ,1 0 2
- a n d g l y c e r o l ,3 6 6 , 7 1 4
- and organization, 32
- hemorrhagiparousagents,448
- in shock. 226
754
INDEX
Decarboxylation,633
Decyl alcohol, 602
D e f e n s e ,l 7 l - 2 2 4 , 3 1 2 , 6 3 9
- adrenal index. see adrenal
* and allergic reaction, I 80
- a n t i b o d i e sl ,E 0 , l E l , l E 2
- a n t i g e n s ,I 8 9
-antiheterogenous reaction,177
- and cancer, 212
- at cellular level, 206
- role of corticoids.640
- d i p h a s i cp h e n o m e n u m ,1 7 2
- a n d d u a l i s m .2 2 3
- f a t t y a c i d s ,6 3 9
- a n d g r a f t s ,2 1 4
- and hetero- and homotropy, 224
- a n d h i e r a r c h i cl e v e l s ,1 8 4
- and infectious diseasc,195
*and lipids,176
- n o x i o u s a g e n t s ,1 7 l
-oscillatory movement, 223
- prolonged hemoshock, 176
- phylogeneticdevelopment, 222
- a n d r e t i c u l o e n d o t h e l i asl y s t e m ,2 2 3
- t i s s u e s2, 0 6
- and trauma. 640
D e f i n i t i o no f I i p i d s , I l 4
Degradatedproteins, 172
Deoxycorticorticosterol convulsive seizu r e s ,3 6 7 , 6 5 9
- defcnse.640
- and glycerol, 367
- a n d h e p t a n o l ,3 E 3
- as lipid,637
- a n d S . d . c .p H , 6 0 4
D e r m a t o t r o p i cv i r u s e s ,2 9 9 , 3 1 4
D e s a t u r a t i o no f f a t t y a c i d s , 1 2 9 , 1 3 0
- in the liver,619
- a n d r a d i a t i o n ,2 3 E
D e s m o l y s e6, 4 2
Destructionof thrombocytes,magnesium,
3_r5
D e s t r u c t i v ei n f e c t i o n , 2 9 5
D e t e r g e n t sa n d c a n c e r ,5 5 0
D e t o x i f i c a t i o ng, l u c u r o n i ca c i d , 6 4 3
- sulfuric acid, 641
Dewar structures,benzene.E
D e x t r o r o t a t o r ya m i n o a c i d s , 1 7 0
a n d c a n c e r ,5 5 1
D i a c i d f a t t y a c i d s ,6 1 8
Diagnosis of pain pattern, 364
Diagnostictestsin cancer,265
D i a r r h e a ,8 3 , 1 6 6
1 , 2 , 5 , 6D i b e n z a n t h r a c e n e
and quenching
of fluorescence,698
3 : 4 : 5 : 5 d i b e n z c a r b a z o l e2, 7 3
9 - , l 0 - d i c h l o r o s t e a r i c4, 0 4
D i c u m a r o l ,6 0 4
Di-epioxides2
, 78
Diethenoics, 3 l5
Diethylaminoethanol, 393
Diethylcarbinol, 122
Diethylene-glycol,3E7
Diethylstilbestrol, 140, 276
Differentiation of cells and cancer. 102.
291,551
Digitaline, 604
9-, l0-diiodostearic acid, 405
Dimercapto-propanol,333
Dimethanesulfoxonoxyalcanes,
27E
Di methylaminoazobenzete,275
2 : 4 Dinotrophenyl-ethylcneimine,280
Dinucleophily, 144
Diols, 387
D i p h a s i cp h e n o m e n o n ,1 7 2 , 1 7 3
- - in hemoglobinuria a frigore, 674
D i p h t h e r i a ,1 8 7 , 1 9 9
Dipolar lipidic boundaries, 126
Dipolarity of red cells in shock, 233
D i p o s i t i v ep o l a r i t y , 1 3 9
Direct action of radiation, 257
- transfusionsand shock, 226
Discharge,electric,545
D i s p l a c e m e n to f t h e d o u b l e b o n d , 1 3 5 ,
622
D i u r e s i s .1 6 6
- and lipids, 162
- and mercury, 402
DNA in viruses,690
Dodecyl mercaptan,333
Double bonds,l3l
- - c h a n g e s ,6 1 9
--conjugation,
132, 135,695
- - fixation on, 622
- - position of, 618
- - a n d s y n j u g a t i o n ,2 E 3
D u a l i s m ,3 5 , 4 3 - 1 0 3
- a b n o r m a l s u b s t a n c e s6, 0
- acid-basebalance,45
- a c i d i f y i n ga n d a l k a l i z i n ga g e n t s ,5 2
- in blood analyses,86
- a n d c a n c e r p h y s i o l o g y ,9 9
- a n d c a r d i a cr h y t h m , 1 6 4
- a t c e l l u l a r l ev c l , 8 2
- a n d c h l o r i d e e x c r e t i o n .6 l
- for coma and convulsion, 163
- in defense,223
- in dyspnea, 77
- of elements, 105
- a n d e o s i n o p h i l e s8. 6 , 5 6 7
- of fatty acids and aoti-fatty acids. l4E
- of fundamentaloffbalances,9l
- in impaired hearing, 73
- for intestines,152
INDEX
'7
55
2s6
E n d o t o x i n ,l 9 7
Energeticcentersof nonpolar group, l3l
- - i n s t c r o i d s ,6 3 5
- factors in carcinogenesis,270
-spectrum of carcinogens,2E8
E n o p h t h a l m i a ,1 0 2 , 2 2 6 , 3 7 1
E n t i t i e s ,l 2
Environmental influencc, 649
- - and surface tension, 65 I
E n z y m a t i ch y d r o l y s i s ,1 7 4
- - of procaine,393
- K n o o p b e t a o x i d a t i o n ,1 2 9
E o s i n o p h i l e s8 ,6 , 1 0 2 ,1 6 6 , 3 2 4 , 5 3 2 ,5 6 7 570
E p h e d r i n e3, 9 1
E p i c h l o r o h y d r i n ,3 4 5 , 3 4 8 , 4 2 2 , 4 3 2
- i n c a n c e r ,5 2 3 , 5 3 3
Epidemiology of poliomyelitis, 298
Epileptic seizures,magnesium, 355
Epinephrine,39l
E p i t h e l i ah e a l i n g , l 6 l
Epoxides,409
- c a r c i n o g e n s2, EI
- of fatty acids, 132, 327
E r y s i p e l a s1, 9 7 , 2 0 4
E r y t h r a l g i a ,5 5 4
E o n u , r a n d h e p t a n o l ,3 8 3
Erythrocytes,see red cells
- in shock,228
E s s e n t i a lf a t t y a c i d s , 1 3 2 , 6 2 1
- and wheal resorption, 566
E s t e r a s e ,1 7 2 , 6 7 4
Ehrlich ascitestumor, 322, 348, 691
E s t r a d i o l .1 4 0
Ehrlich mammary tumor, 715
E s t r o g e n i cp r o p e r t i e s ,1 4 0
E l e c t r i c d i s c h a r g e s5
. 45
E t h y l a l c o h o l i n c o n j u g a t i o n ,1 2 2
- b u t y r a t e ,6 7 4
Electron. 544
- transfer. 5
- m e r c a p t a n3, 3 0 - 3 3 3
E l e c t r o n i c c o n f i g u r a t i o n .e l e m e n t s , 1 0 6 - - i n t h e r a p y , 4 7 6
E l e c t r o p h o r e t i c a n a l y s i s o f a n t i b o d i e s , - s u l f a t e ,q u e n c h i n ge f f e c t o f , 7 0 9
t8l
E t h y l e n et r i t h i o c a r b a m a t eq, u e n c h i n ge f E l e c t r o s t a t i cf o r c e s i n a t o m , 5
fect, 709
- - in molecules.6. 9
Ethyleneglycol,38T
E l e m e n t s 1. 0 5 . I l 2
E t h y l e n i m i n e s 2, 8 0
- anli-A, 105,349-361
E t h y l e n i m o n i u mi o n , 1 7 9
-anri-D. 105.394-404
Evaluation of projective personality, 308
- i n o r g a n i z a t i o n .1 0 7
-of
r e s u l t si n c a n c e rt h e r a p y ,4 1 7
- p r o p e r l e v e l ,3 5 0
E v e n i n gd y s p n e a ,7 7
E l e o s t e a r i ca c i d . 3 2 0 , 3 3 5
E x c r e t i o ni n d e x . 8 6 , 4 1 5 , 5 9 4
- - and carcinogens.708
- o f s u r f a c ea c t i v es u b s t a n c e s1,6 6
--defense against,639
Exercise and blood changes,67 |
- - and oxidation, 129
E x f o l i a t i v ec y t o l o g y ,4 0 , 5 5 0
--and
S.d.cp
. H,601
E x o p h t h a l m i a1, 0 2 , 3 7 0
E l e o s t e a r i ca l c o h o l .3 8 8
Exotoxin,197
Embryos' unsaponifiable fractions, 371
Experimentalcarcinogenesis,see carcinoE m u l s i o n .1 2 4
genesis
E n c e p h a l o i dt u m o r s , 3 7 2
E x p l o r a t o r yp s y c h o t h e r a p y3, 0 E
756 /
lNoex
Extracellularpotassium,547
Extrasystoles
and lipoids,661
rNDEx
F l e x i b i l i t yo f f a t t y a c i d c h a i n . 1 3 7
F l o c c u l a t i o n ,1 7 9
F l u o r i n e , 1 0 5 ,4 0 4
F l u o r e s c e n c es, e e q u e n c h i n g o f
F o c i . c o n s t i t u t i o na n d p r o c e s s e isn . 6 0
F o o t u l c e r a t i o n sa n d c h o l e s t e r o l ,3 7 1
F o r m i c a c i d , l 2 - s ,- 5 4 5
Fractionation of f atty acids, 627
F r a c t u r e sa n d a c i d - b a s eb a l a n c e , 5 0
Free lipids, 622
F u n c t i o n a lr o l e o f f a t t y a c i d s ,1 3 0 , 1 3 5 ,
363
.3
F u n d a m e n t a lo f f b a l a n c e s 9
CrLrclosrvrNp, 145
C a l a c t o s e ,1 4 5
G a l l b l a d d e rc o l i c , 5 0 , 4 3 2
G a m m a g l o b u l i n s ,1 8 3
- r a d i a t i o n ,2 4 1
G a s e o u sg a n g r e n e ,1 9 7
G a s t r i c u l c e r a t i o n sa n d c h l o r i d e s ,4 0 3
C e n e s ,p l a c e i n o r g a n i z a t i o n ,2 5
s ,0 2
G e n e t i cf a c t o r i n c a r c i n o g e n e s i 3
Geriatry, see old age
Glioma virus, 295
C l o b u l i n i ca n t i b o d i e s 1
, 83
G l o m e r u l o n e p h r i t iisn i n f e c t i o n ,2 0 4
G l u c a g o na n d z i n c , 4 0 I
Gluco group, 145
G l u c o n a t ei o n a n d S . d . c . p H , 6 0 0
G l u c o n i ca c i d . 1 7 0
G l u c o s a m i n ei n a r t h r i t i s , 4 3 5
- a n d c o n n e c t i v et i s s u e ,4 3 E
- a n d i r i d o c y e l i t i s4, 3 9
- s y n t h e s i s1, 4 5 ,6 3 8
G l u c o s a m i n i ac c i d ,6 1 8
G l u c o s ea n d c o n v u l s i o n s 3
, 6E
- a n d h e m o r r h a g e ,4 4 0
- o x i d a t i o na b n o r m a l , I 3 2
- pH of tumors,368
- a n d p o t a s s i u m3
, 98
- a n d S . d . c .p H , 3 6 E , 6 0 3
G l u c o s i d e s6, 0 4
C l u c u r o n i ca c i d , 1 3 2 , 1 4 7 , 1 7 0
- - c o u p l i n gm e c h a n i s m 6
, 41
- - a n d s u l f u r i cr a d i c a l s5. 4 2
- - in urine,86
G l u t a t h i o n ,3 4 1
C l u t a m i ca c i d , 1 6 9
G l y c e r i ca c i d a n d a l d e h y d e ,3 6 8
s ,5 8
G l y c e r o la n d a r t e r i o s c l e r o s i 4
- c a l o r i c m e t a b o l i s m ,3 6 3
- a n d c a n c e rt h e r a p y ,4 9 3 , 5 3 3
- c a r d i a cr h y t h m , 7 1 4
- c h a n g e si n d u c e s .3 6 6
-and chills,366,713
- a n d c h o l e s t e r o l3, 6 i
757
G f y c e r o fc o n v u l s i o n s7,1 4 , 7 l 5
- a n d d e s o x y c o r t i c o s t e r o naec e t a t e ,3 6 7
-and fatty acids,135
- h e a l i n go f w o u n d s ,3 6 5
- and hemorrhage4
, 40
- h y p e r t e n s i o n3, 6 6
- a n d m o n o m o l e c u l alra y e r ,3 6 3
- pain, 364, 422
- a n d p a t t e r n s 3, 6 8
- a n d r a d i a t i o nl e s i o n s .3 6 - 5
- a n d s a t u r a t e df a t t y a c i d s , 3 6 2
- and S.d.cp
. H, 603
- a n d t r a u m a ,4 2 4
- and tumors,365
- a n d v i r u s e s ,3 6 4
G l y c e r o p h o s p h o r i ca c i d a n d c a n c e r , 5 3 3
- - b o u n d t o f a t t y a c i d s , l 3 0 , 1 3 7 ,3 6 2
- - and postoperativepain, 426, 723
- - a n d S . d . c .p H , 6 0 3
- - t h y r o i d e f f e c t s ,3 6 9
G l y c i n e ,- 5 4 5
Clycogen in foci, 60
G r a f t s o f t u m o r s i n h u m a n s .2 1 4
G r a m p o s i t i v i t ya n d l i p i d s , 1 5 7 , 3 6 9
G r a n u l o c y t e sa n d h e m o s h o c k , 1 7 2
- l y s i s ,1 7 4
G r a s s - t e t a nay n d m a g n e s i u m 3. 5 5
G r o w i n g t u m o r a l m a s s ,4 0
C r o w t h a n d b u t a n o l ,3 7 9
- i n d u c i n g f a c t o r a n d p o t a s s i u m ,3 9 6
. 65
G u i l l a u m i ni n d i c a t o r 5
(iumatous and iodine, 404
H r l o c r x l c c o m p o u n d so f f a t t y a c i d s ,3 2 8
Headaches,422, 423
H e a l i n g p r o c e s sa n d b u t a n o l , 4 2 2
- - o f t i s s u e sl,6 I
- - a n d p o l y u n s a t u r a t e da l c o h o l s , 4 2 6
- - postoperative,422, 426
- - r a d i a t i o n l e s i o n s ,3 6 5
-a n d u n s a p o n i f i a b l ef r a c t i o n s , 3 7 t
- - wounds, 365
H e a r i n g i m p a i r m e n t a s a c i d - b a s es y m p tom, 6l
- - p a t t e r n s7
. 3 , 4 3l
- - t h e r a p e u t i ce s s a y s 4, 3 0 , 7 2 4
Heart, see also cardiac system
- b l o c k a n d l i p o i d s ,6 2 2
- a n d u n s a p o n i f i a b l ef r a c t i o n , 3 7 2
H e a t i n a c t i v a t i o no f v i r u s e s ,2 9 4
- t r e a t m e n to f f a t t y a c i d , 7 0 0
H e a v y e l e m e n t s3, 6 1
H e m a n g i o m a t o utsu m o r s ,4 0 1 , 6 5 3
H e m o c l a s i a1. 7 2
H e m o c o n c e n t r a t i o inn s h o c k ,2 2 8
Hemoglobin and coppcr, 359
758
tNoex
H o m o t r o p i c e l e m e n t s ,1 0 5 , 3 4 9
- processesand ulceration, 224
H o r m o n e s a n d c a r c i n o g e n e s i s3,0 1
Horse radish peroxidase,409
Human placenta,see placenta
H y a l u r o n i d a s e6, 4 9
H y d r a z o d e r i v a t i v e s2
. 75
Hydrogen peroxide, 345
H y d r o i d s ,I l 9
Hydrolytic producls, 192
- d i g e s t i o ni n s h o c k , 2 2 8
- e n z y m e s ,l 9 l
- rancidity, 128
H y d r o p h o b i cg r o u p , I l 9
H y d r o n a p h t h a l e np
e e r s u l f i d e s4.9 l . 5 3 3
H y d r o p e r o x i d e s1, 2 9 , 4 0 8 , 4 0 9 , 6 1 7
- of fatty acids, I 3 I
Hydrophilicgroup, I l9
H y d r o p e r s u l f i d ei n c a n c e r t h e r a p y , 4 8 t ,
533
- a n d m i c r o b e s .3 3 4
- and pain,422
- a n d p a t t e r n s ,3 3 4
H y d r o s p h e r e ,5 4 7
Hydroxonium ion and resonance8
.
Hyperalgesia,554
H y p e r k a l e m i a3, 5 3 , 6 1 4
H y p e r c h r o m i aa n d o f t b a l a n c eA , 3 9 9
H y p e r l e u c o c y t o s i st,6 6
H y p e r t e n s i o n1, 6 6 , 1 7 2 , 1 7 3
- _ a n d a d r e n a lf a t t y a c i d s ,3 7 1
- a n d g l y c e r o l .3 6 6
Hyperthemia,| 66
H y p o c h r o m i aa n d o f f b a l a n c eD , 3 9 9
Hypokalemia,39T
H y p o t e n s i o n ,1 6 6 , 2 2 6
Hyponatremia, 397
Hypothermia,166, t74, 226
H y p n o t i c s ,6 0 4
I p r v u x e a n t i b o d i e s1, 8 2
- s e r a . 1 9 8 , 1 9 9 ,2 0 1 . 2 0 3
- s t a g e o f d e f e n s e1, 8 3
l m m u n i t y a g a i n s tv i r u s e s .2 t 8
Immunological aspectof cancer, Zl?-220
- - a g a i n s tc e l l s a n d t i s s u e s 2, 0 6
-t h e r a p e u t i ca p p r o a c h ,2 l E
I m p a i r m e n to f c i r c u l a t i o na n d s h o c k , 2 2 6
I n c r e a s e dp e r m e a b i l i t yi n s h o c k . 2 3 2
Incubation time in allergy, 185
- - i n f e c t i o u sd i s e a s e s1, 9 . 5 .1 9 6 , 1 9 7
I n d e x o f e x c r e t i o na n d r e t e n t i o n ,5 9 5
- of repartition of anesthetics,125
Indigorubine,89, 593
I n d i g o t i n ,8 9 , 5 9 3
I n d i v i d u a l i z e dt r e a t m e n r .4 l 3
Indoxyl in urine, E6, 89, 132, 591
INDEX
lnduced carcinogenesis,
268, 290, 703
- - and conjugated fatty acids, 703
- convulsions,see convulsions
- shock, 679
- tumors, see carcinogenesis
Infection and abnormal fatty acids, 319
- destructive,295
- neoplastic,295
Infectious disease, allergic and toxic
p a t h o g e n e s i s1, 9 5
- i n c u b a t i o n ,1 9 6
- s y s t e m a t i z a t i o no f , 1 9 6
Inflammatory form of breast carcinoma,
215
I n o r g a n i c a c i d s a n d S . d . c .p H , 6 0 2
I n s o m n i a8, 3 , 1 6 6 ,3 7 3
l n s u l i n a n d p o t a s s i u m ,3 9 E
-and
zinc, 401
Intake of food and itching, 62
and pain, 45
I n t e n s i t yo f s t i m u l u sa n d i t c h i n g ,5 6 2
I n t e r m e d i a r yl y s a t e s ,1 9 4
Internal sea, place in organization, 30
I n t e r s t i t i a lf l u i d s a n d e l e m e n t s ,1 0 7
- - p H a n d p a i n ,5 5
I n t e r v i e w so f c a n c e r p a t i e n t s ,3 0 8
I n t e s t i n a l a b s o r p t i o no f f a t t y a c i d s , 1 3 5
- 5sg1s1i6n5
in shock, 230
I n t e s t i n e sa n d l i p i d s , 1 6 2 , 3 7 2
I n t r a c e l l u l a rp o t a s s i u m ,5 4 7
- v a c u o l e si n s h o c k . 2 3 2
I n t r a c t a b l ed i a r r h e a ,3 7 2
I n t r a c y t o p l a s m a t ivci r u s e s ,2 9 0
I n t r a n u c l e a rv i r u s e s ,2 9 0
- vacuoles in shock, 232
I n t r a v e n o u si n j e c t i o ni n d u c i n gs h o c k ,2 2 6
I n v a s i v ep h a s eo f c a n c e r ,4 0 , 3 9 6
- - a n d p o t a s s i u m ,3 9 6
- - a n d s m e g m a ,3 0 5
- - a n d s t e r o l s ,3 0 6
I n v a s i v e n e sasn d c a l c i u m , 3 5 7
I o d i n e . 1 0 5 .4 0 4
I o n i z i n g r a d i a t i o n ,5 4 5
I r i d o c y c l i t i sa n d g l u c o s a m i n e4, 3 9
I r o n , 1 0 5 ,J 9 9 , 6 0 0 , 6 0 4
l r r a d i a t e dw o u n d , 2 5 1
I r r a d i a t i o n ,s e e R a d i a t i o n
Irreversible fixation of chlorides, 134
I s a m i n eb l u e a n d h e m o r r h a g e ,4 4 0
Ischemic infarct and cholesterol,370
Isotropic resonanceforms, 302
Itching as acid-basesymptoms, 6l
- a n d i n t e n s i t yo f s t i m u l u s ,5 6 2 , 5 6 3
- pathologicaa
l n d p h y s i o l o g i c a l6, 1 , 6 2
Krt-eltrr, 166
K a r y o r r h e x i s ,8 2 , 1 6 0
7s9
760
INDEX
rNDEx
761
762
INDEX
Mitochondria, 126
- m e m b r a n e .6 1 6
- pre-ferments,223
Molecular oriented layers, 123
and glycerol, 363
Molecules, forces, 6, 9
Molybdenum, pharmacological activity,
399
Mono ammonium phosphateand pain' 54
Monocellular organisms, 3 l2
- - place in organization, 26
M o n o e t h e n i cf a t t y a c i d s , 3 1 3
position of double bond, 618'
619
M o n o - g l y c e r i d e s1, 3 5
M o n s t r o s i t i e s ,1 5 9
- and carcinogens,268
- and ontogeneticallotroPY,303
Morphine and hemoglobinuria a frigore,
679
-and
sbock. 1'14,229, 670
Morphological changesas pattern, 159
Motion of particles in organization, I I
Multiple carcinogenesis,270
Multiplication and organization, 3l
- of viruses, 690
Mumps, 197
M u s c u l a re x e r c i s ea n d l i P i d s ,l 5 l
--and
s h o c k ,6 7 1
Mutations and carcinogens,268
- ontogenetic allotroPY, 303
Myocardial infarct, 7th day manifesta'
tion, 2l I
- - and oxygen, 407
- insufhciency and unsaponifiablefractions, 372
Myristic acid, 639
Myxoma virus, 295
NrpxrsrLexe perselenide,344
--and
, 1 9 ,5 3 3
C a t h e r a p y5
2-naphthylamine, 273
Narcotics, 604
- and anti-hemorrhagic action, 448
N a s a l p H , 8 3 , 1 0 2 ,5 6 5
Nature, dualism in, in, 35
- o s c i l l a t o r Ym o v e m e n ti n , 3 4
Neck pain, 422
Necrosisof tumors, 160, 3E7
Negative phase in heart physiology, 660
--in
d e f e n s e 1. 7 3
Neoglucogeniccorticoids, 144
--and
coma,659
- - and conjugated fattY acids, 362
- - and convulsions,659
- - and defense. 640
-a n d g l u c o s a m i n e6, 3 7
l. 2. OcrlxeoIor, 387
l . 8 . O c t a n e d i o l1
, 23
Octanol. 380, 382, 602
Oestral cycles and lipids, 164
Offbalances. 93
OffbalanceA and agents,4l I
- - analyses,95
- - and arteriosclerosis,453
-a n d c a n c e r ,9 9 , 5 3 2 , 5 3 5 , 5 3 7 , 5 4 0 ,
546
--and
e l e m e n t s1, 0 5 , 3 4 9 ,3 5 3 , 3 9 4
- - manifestations,102
- - a n d s t e r o l s ,3 7 1
- - trauma. 423
- - w o u n d s .2 1 4
- - seealso specificagentsand analyses
Offbalancc D, agents,4l I
- - analyses,95
71)
rNDEx
O f f b a l a n c eD a n d c a n c e r , 9 9 , 5 3 2 , 5 3 5 .
537.540
--and
e l e m e n r s1, 0 5 .3 4 9 .3 5 3 .3 9 4
- - a n d r a d i a t i o n ,2 6 1 , 6 8 3
- - a n d t r a u m a ,4 2 3
- - seealso specificagentsand analyses
OH as polar group, 120
Old age and lipoids,646
procaine,393
w h e a l r e s o r p t i o nt i m e . 4 1 9
O l e i c a c i d , a d r e n a l d e f e n s e ,6 3 9
--bacteriophages,3l3
- - d e r m o l r o p ev i r u s e s .3 1 4
- - a n d o x i d a t i o n .1 2 9
O l e i c a l c o h o l .3 8 8
O l i g u r i a .8 3 , 1 6 6
Ontogenesis,24
O n t o g e n e t i ca l l o t r o p y , 3 0 3
O p i u m a l k a l o i d s .2 2 9 , 6 0 4
O r g a n i c l e v e l .2 3
- - a n d c a n c e r ,4 0
- - d u a l i s m ,8 3
--elements, ll0
- - m a n i f e s l a t i o n s1, 6 2 , 1 6 3 , 1 6 4
O r g a n i s ml e v e l , s e e s y s t e m i c
O r g a n i z a t i o na n d a n t i e n t r o p y ,3 , 5
- atom, 3
- basic concept, ,l
- b i o l o g i c a lr e a l m , l 7
- i n c a n c e r ,J 8 - 4 2
- a n d c a t i o n s ,2 7
- a n d c o n s t a n t s3
.3
- a n d d e a t h .3 2
- a n d h i e r a r c h i ce n t i t i e s ,1 2 . 2 0
- o f i m m u n i t y ,1 8 6
- a n d l e v e l s ,I 2
- and life, 32
- m i c e l l e s ,l 0
- m o l e c u l e s ,6
- a n d m o t i o n o f p a r t i c l e s ,t I
- a n d m u l t i p l i c a t i o n3, l
- a n d p h y l o g e n e t i cd e v e l o p m e n t
,24,222
- a n d p o l y m o l e c u l a rf o r m a t i o n s ,9
-social, 19,548
- and solar energy, 33
- s u b a t o m i c ,I I
- s u b n u c l e a r5, 4 3
- a n d w a t e r c i r c u l a t i o n .3 0
O r g a n i z a t i o nr o l e , f a t t y a c i d s , | 3 5
-g l y c e r o p h o s p h o r i ico n , 3 6 3
O r g a n i z e db o u n d a r y , l 5
O r i g i n o f a l l o p r e g n a n e3
, 61
- o f a m i n o s u g a r s6, 3 6
- o f c h o l e s t e r o l ,1 3 9
O r i e n t e d ,m o l e c u l a rl a y e r s ,1 2 3
O s c i f l a t o r ym o v e m e n t ,3 4 , l 7 Z
- - a n d d e f e n s e .2 2 3
763
Osteo-arthritis,450
- - pain and acid-basebalance,50
- - t h e r a p y ,4 3 5
O s t e o m a l a c i aa n d c o p p e r , 3 5 9
- a n d m o l y b d e n u m ,3 9 9
O t o f o g i c a lc o n d i t i o n s ,6 7 , 7 3 , 4 2 8 , 4 3 0
Oxalic index, 23E, 6E0
- a n d r a d i a t i o n ,2 4 0 , 2 4 5 , 2 S l , Z S 3 , 6 8 2
- and shock, 227, 248
O x i d a s ea n d a m m o n i u m m o l y b d a t e ,3 9 9
O x i d a t i o n ,a b n o r m a l , 1 3 2 , 1 4 7
- o f f a t t y a c i d s ,1 2 9 ,l 3 l , 1 3 3 ,3 5 4 ,6 1 6 .
6r8
O x i d a t i v ed e s a m i n a t i o n3
, 54
- fission of fatty acids, see oxalic index
O x i d i z i n g s u b s t a n c e ss, e e a l s o p e r o x i d e s
* - a n d r a d i a t i o n ,2 5 4 , S 2 S ,i 9 4 , 6 g 3
- - a n d s e l e n i u m3, 4 3
- - in urine,89, t}Z, l3Z.265-266
Oxidoreductionpotential of foci, 59
- - o f u r i n e , 8 6 , 8 9 , 1 0 2 ,5 9 0
O x y g e n ,l 0 E , 3 4 5
- and anoxybiotic process,407
- and conjugatedfatty acids,700
- fixation of, 156
- and pain pattern, 405
- sedativeeftect of, 407
- t h e r a p y ,4 0 6
- t r a n s p o r t ,6 5 7
P r r x , 4 3 - 6 1 8, 3 , 1 0 2 ,3 1 3 ,3 1 4 ,3 1 6 ,3 t 7 .
380,421,532,552
- see also specificagents
- a n d a c i d - b a s be a l a n c e4. 6 . 5 6 1
- a m m o n i u m c h l o r i d e .5 4
- i n a r r h r i t i s4
, 35
- and bixin, 324
- a n d b u t a n o l ,3 8 0
* c o n i u g a t e df a t t y a c i d s ,3 2 2
- different agents, 422
- d u a l i s m ,4 3 , 4 5
- ethylmercapta3
n3
,1
- a n d g l y c e r o l ,3 6 4
- h e p t a n o l ,4 2 2
- hydroperselenide,344
- h y d r o p e r s u l f i d e3, 3 4
- and intake of food, 45
- l i p i d sa n d l i p o i d s ,1 6 0 , 6 5 7
- and local pH, 55, 5E
- m o n o - a m m o n i u mp h o s p h a t e5, 4
- o s t e o a r t h r i t i s5.0
- a n d o x i d o r e d u c t i o np o t e n t i a l ,5 9
- and oxygen, 4O5, 407
- p a t h o l o S i c a l4, 4 , 4 7 , 5 5 3
- p a t t e r n s ,4 7 , 5 3 2 , 5 6 1
- p e r s e l e n ied, 3 4 4
- physiological,44, 552
764
INDEX
P e r o x i d e s ,1 4 6 , 4 0 9
-of
fatty acids, 129, 132. 327
- and iodometry, 593
- a n d r a d i a t i o n ,2 5 4 , 5 2 5 , 5 9 4 , 6 E 3 , 7 2 6
- a n d s c h i z o p h r e n i a ,4 1 9 , 5 9 4
- a n d s e l e n i u m ,3 4 3
- in urine, 86, 102, 132, 254, 265-266.
525, 592
P e r s e l e n i d e s3,4 4 , 5 1 9 , 5 3 3
P e n u f f i d e s3
, 3 6 , 4 2 2 ,4 9 1 , 5 3 3
Pertussis,197
P e t e c h i a e2, 2 8
pH, see also S.d.c. pH
-local,55, 166,316
- t u m o r s a n d g l u c o s e ,3 6 E
- urine. 90, 532, 557
Phagesand arachidonic acid, 316
- a n d g l y c e r o l ,3 6 4
- a n d l i p i d s ,6 5 6
- see also specificagenls
Pharmacology, see pharmacodynamic
activity
Pharmacodynamic activity
- - 3lnsrmal fatty acids.3l9
- - a c i d l i p i d i c f r a c t i o n s ,3 1 7
- - agents with negative polar group.
3t2, 329
- p o s i t i v ep o l a r g r o u p , 3 6 ? , 3 1 , s
- - a l c o h o l s ,3 7 5 , 3 8 0 , 3 t 9
- - a l p h a h y d r o x y f a t r y a c i d s ,3 2 6
t h i o f a t t y a c i d s ,3 3 E
- - a n t i f a t t y a c i d s c o n s t i t u e n t s3, 6 2
- - a n t i o x i d a n t s4, 0 9
- - arsenic,bismuth, mercury, 402
- - b i x i n e ,3 2 4
- - b u t a n o l ,3 7 5
- - c a l c i u m ,3 5 4
- - c a t i o n s ,3 9 5
- - chlorine, fluorine, 403
- - c o b a l t ,3 6 0
--compounds
with thiol groups, 33_5
- - copper,358
- - c o r t i c o i d s ,3 7 2
- - diols. 3E7
- - e l e m e n t s3, 4 9 , 3 6 1 , 3 9 1
- - epichlorohydrin, 348
- - fatty acids, 3 12, 326
- - glucose, glycerophosphoric acid.
368
- - g l y c e r o l ,3 5 3
- - halogencompounds fatty acids, 328
- - heavy etements,361
- - heptyl- and bexyldisclenide,343
- - hydropersulfides,334
- - iodine, 405
- - iron, 399
- - Iipoalcohols, 388
INDEX
765
766
INDEX
Ir-DlX
It:rdiali*n lntl trriilir inrJlx. l.i,t, l{i). l+5
-<l
'(i
,,h1
. - - ; r n r Jp u i r r . 1 5 . 1
- ;rnd fotritl\. I lt,. :s:
f
- t t l i g r ; t l i o r rl.5 ( l
' ' - l ) f i r lt t n r l r p r : n i l i l r l r lfct ; r c lj r r n r , : . i h j . l
4tv
-
3 p d u r i n { i t n r r l r , r c1r ,5 . . 11.5 . s l, { r l . l 6 l
5:J. -r:5.f;Nl. itf)
- . * : r n r l! l ! c L l l . l rl c s i * n r .: 5 J . 1 5 6
- ' . a n t l r i n c . . 1 {I}
R a d i l l e t j v e r r r r . l i i t m ,a { 1
* - - \ o l l t C C \ .1 { |
R ; r r l i o r r r i n r e l iic: l [ e t l r r f f . r l l r . ; r c i i l r , ] { g
R ; r t l i o t i r c r , r p i J. : J . ( ) 1 1
. i r i r r i r r g i r : i t l gi ru i r l c r l .l { r I
- - - i r n d c h r . r n r r l l l ' t r , i l t n i\ l.f ,
R ; r ' l i r t r rhrr r r n : ,l : 1 . J l ( ) " 1 1 - - * i l t n r c n c l r r r et , r l , l J { 1 , : . . l : , : { l
- In
n l i r t i n l l n t:.. 1I
R r t n * i r l i ! ! ' i. r h p l i l1 1t11 f l r l l r . r ,j,i:l r , I t 1
. t t f c d i h l r f ; r t r , - 1I t t
o f l l l t l ' l r c i i l s ^l l H
. -'
fl(n)\irlit\0. 119
R ; r p i r rl r g i n gl r n J c r r l c i L r nir .\ l
- . c e l l u l r r r ,l l ' : . l h a l
R ; t t r ; r t i . ' o r t r;l rr n r le t h r l r r r c r t , r p l l r n .i 1 , 1
l t * ; r c ti r ' ; r il o n t ' f r i r L r r r s . l ( ) J
Rcclurnlrnd lrPiiir.lfrl
I t e c u r r c n t f r . rc r . l t ) j
R ec l c c l l r , f r r t t r r c j r l s . I I i I l t i . f r h l . f r ( r , l
fr 6.-i
I t n t l l t p i t ' l . l r , r r i ' r p l rl . { r { r f r
1 1 n l l\ \ \ \ ! e n t i , t t ) . i r ( )Lt l ) f , irtlrl J)()lit\\nttil,l, I I
l(!illr';rll\ il1l!l \lll(l!f:. {r-ft
r t ' r l i t t t c t t l i r l i r r r . tl r 1 r . S { r . ! f r { , . { r i l ri
itnrl rtr:tillr. l{rtl. i-(i. .l{<. /,i,i
fr (r il
r a c t l r l l ; tilo t t . ( ' { r . J
Rcdu,.'rd rrrctirlr. 19(.)
RcItn'-'talillr of lircr. lt, I
r r n r lt : l l c i r i r n . i 5 l ,
;rnrllipitlr. l{':. (ril
I t e l l r l i o n r h r pl r t . t \ \ u e . !i li l r i i c ( ) l d \ . { ) i :
R c n a l a c t i r . er c ; r h r o r p t i l n . ! f )
Renul ct:lic. "lJl
R c n i r l c . r c r et i o n , 9 { }
R c p c l t t c l p l r \ \ ; r S fu n r l v i r r r r p ; r t l r o g c r r r c i l l
:!5
Rcurttlttcc fornrr, li
-.- --- irnd ullolnrl]r'. ltii
-- *- hcnrcnc. X
** - clrrho.rt,l. li
. - - h r d r n . t o r t i u r r ri r r r r .S
- ' ' i t t c r r n c * l p i r t h r . r g c n * r i : .3 0 1
761
R r r r J r : r r ]r I i i i f ! c . : " { - i
ll i r,rtr,.rCC
t.tCC,fr(lfi
R r r p i r i r l $ r \ ( ) \ ) h i ( ) t i L n h : 1 \ ei r t l t l f r r c i , f r ( l
R r r L lrt : . e r ' , r l t r . r l i c on f , J 1 ?
[ t r l e r ]l J ( ) n1 1 1 1 1 ci )1{.1 .. 5 9 {
R l : \ r r n i l c i t r r l u s l s l t r l".1 5 { r
r n . l c l ' q n r e .l l J
R l t c r r n r i r l i ul r r ' * r . l t l l
R l t c t r r t t , i l o i rul r t h r i t i r , . 1 . 1 - 5
R h i n r r . p h ; r rnr g c l r l i n f r c t i q : r n i". { 11
R l r rt h r r r ,c ; L r t l i ; r ch,5 9
il1 nirlltrt.,1J
R l l r l n r r r ' l r . i ui r e i r l r n ( 1 t g i r n i . 1 a l i o n: l,l
R l c t t t , r l r ' r ;ur l c o h r r l .l N l { . l l { 9
[ { r i r r t o ] r ' i c1 r ' i g l r e c r i r i e 1. . 1 ( r
l l r ' l , r ' l l r t . r .| ( ) l
R i s i , l i i r i r f t l t r . 'r . ' r . l r c r i r l r i r r . l . t ?
I { r r l t ' r i [ . r t tl r n l r l ' t t r t l , : f r : n s c{ ,r J l {
ilt rillllltlr0rt.l.\!)
n r r eg c l l c r i i t r o n , I f r l , 6 . l l {
r . i j r r L u l p u r r r r r : ; r h i l i nI .l 6
! ( ) l j \ l : i t l l \i t t , ' l q 1 1 1 1 1 7 . , 1] f; t r r t .
l ' . i t l ! ; r u r r l r .I l r r
lipitl' in lrlo,rtlphlrirrl.g1,. 6f:)
I t { r i r l r ' ;jrr{ l o r n t i r t i o r r t r r r r ,lli p r d : . 6 ? 1 1
irr .hoiln.lJ 1
I i r r l \ \ , t t ! ( ) n f : ]r 9. 5
R u h h er h l r . l r u p er o r , i t l t r . l l 9
l l r r h i r l i r r n rl ,t ) 7 , 3 9 l l
5 r t t I o \ \ I H r r i l l i p o - ; s l g 1 y | 1 ,11311. 1
,
\ l r l t t ; i t r I i t r t t r l ; r n t j r ; r f ' r e r .l t ) ( )
\ . r l i c . , ' l i rr r l . i c hrrl c r r n r lp l i i r r .J 3 l
\ . r p , r 1 1 1 l i ; l l rl l1gt - l r , r t 1 r ,r r , ' t l i p r r l r C t r , tl r. n t l
: r er r l l i p t r l i , , f ' l] a i t | t t l
\ . r l r , ' r t t t t r "{ ,r { ) " 1
\ . r r i o r i l ; r i i r i r l{ l t l r f i r a t r t 0 f l t r n r t r t l . I l f r
l i , u l \ l ( r f l t l . l t l ( )onl l t t t t t t r t r J. f l
\ . r i t ; r , r l r " tfi, r l l i i i L r r l r _t ; L j r i r i ur o l c . l : \ i
r l r l e n r c . l ! " r t n \ t .6 3 9
' ' ' ' '- . ; r n r lg l l u c r , r l . . l f r l
p l t ; i r n l ; r e o l t i g l .1 l l
.
r l t r i r n r i r i i l , , ,l l N
i r n d S . r , . r .1 1Il . 6 { ) 1
\ . r l l i t . r t r r r inl l ( l r l c \ . r t r i r i r t i r ri rnr l i r r r . f r I r J
i ; { ; r r f r ) r { l } i 1 1 rgl [ r : c t . r ; i d i l l i i : n . ] - ! 6
S u h i z o p h r c n i i r. r n ( l h u t . r n r i l ,7 l E
' - - ; r r r t lr l r i n ( '
I l e t o r j d c s . ^ ll , 4 t 9
\r-lcrotir rcrir in hclring impirirntcnt, ,i)
S c r l en r n g p r ( ) j c { l i n c i r n c c r . - 1l ?
\*.<rntl rltrl rv*untl crutt Fll. l{}J. 'l 11.
l : l { . 1 1 6 . 1 \ 1 . 1 1 { r.}t.a l . . { g l f r l l
\rr \llcciliL irgfn{t
hrttlugiill iticturs. {ri)t{
- - - - c h c m i r l r l i r g en t r , . t 9 9
. i i n r l p h 1 . , : i c ; r ;l t g e n l \ . 6 ( ) :
Srrr iotrotitr^.rrir:n
I l,l l l
768
./
tNoEx
rNDIX
.'.' 769
Squalcneand cholesterol,6.1I
S q u i de x t r a c t s ,6 7 3
S t a p e sm o b i l i z a t i o n .4 3 0
S t a r v a t i o n ,l i p i d s , 6 1 4
Stutus epilepticusand magnesium, 3-56
S t e a r i ca c i d . d e f e n s ea g a i n s t .6 3 9
S t er i c c o u p l i n g ,l 4 ? , 6 3 3
- p o s i t i o n .l 4 l
S t c r o i d s l, l 7 . 1 4 6 ,{ f i 6
. * d c r i v i n g f r o m a r a c h i d t l n i ca c i d , 6 3 |
- ' a n d f a t t y a c i d s .3 1 6
- c n c r g e l l cc e n l e r s ,6 J i
r r i ( h ; r l \ \ ' { r - a i ! ht r r r t s l ' i ; r r n , i . 1 I
1 g q '; s i r r r \ l r L r ' i l i i r t g l ( } t r i s
Slcttiu. rs1 ;1lro cl)r)lf\l(.r{}l
'
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S l r L t t t t i L t r r r1. { } i . : / , 1 . { , 1 ) 1 1
\ r t l . t r , t L r l . ' i l r ' t r n t l \ i , l ; i . i i 1 l. \r i l ! l . , i l 1 ,1. . . 11 r ,
\ r t l ' l r : t l i r ' l l r i , r , . { t , i l r r x ;1l l l , . ! ( } \ i r l r i " r n i l l r .
l'3...
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\ , r , ' t t ' l t t l l , r i . t ) , / i"r l.{ r i
\i. it,,'
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l t . l r , i . r , l r r , l i i r . r . r g .. .r',r , i r
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, r 1 . 1l i p r t i r . l ( . 1. l 6 J
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j hv
l l t r i \ l t . r l r p l c ' , t i r ' r r .{ r ( . 1
! r ' r r r 1 . l , , 1lsr ' ,n r r l l r r r i . . { r 1 f r
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770
INDEX
T r e a t m e n to f a g e n t s ,5 3 3
- - butanol, 493
- - biological guided chemorherapy,
526
- - cod liver oil fatty acids,466
- - conduct of treatment, 536
- - conjugatedfatty acids, 501
- - criteria of, 532
- - glycerol, 493
- - heterogeneousagents,508
- - mercaptans,476
- - p r e s e n tf o r m , 5 3 1 - 5 J 7
- - radiation, 524
- - s e l e n i u mp r e p a r a t i o o ,5 1 2
- - sterols, 465
- - sulfurized oil, 4EI
- - tetralin perselenide,5l E
- - persulfides,491
- - thiosulfates,482
- - unsaponifiablefractions, 468
- - see also Other agents
T r e a t m e n t s ,g u i d e d ,4 1 3
- a g e n t s4
, 16
-criteria,4l5,
- c v a l u a t i o n ,4 1 7
- i n d i v i d u a l i z e d4, 1 3
- in successivegeneralions,7 I 5
- t u m o r s i n a n i m a l s ,4 1 7
- of other conditions than cancer. 4/E
- of acid-basemanifcstation, 420
- o f a l l e r g i cc o n d i t i o n s ,4 5 l
- of arteriosclerosis,457
- o f a r t h r i t i s ,4 3 5
- of burns, 426
- of gallbladder colic, 435
- o f h e a r i n g i m p a i r m e n t ,4 3 0
- of hemorrhage,439
- of itching, 435
- of pain, 42 I
- of trauma, 423
- of vertigo, 42E
Trichlorcthyleoe, 404
Trigerminal neuralgia, butanol treatment.
721
Trichloropropane, 340
Triglycerine compounds, 135
T r i p h e n y l e t h y l e n i ca c i d , 2 7 5
Tryptophane oxidation, 132
Tubercle bacilli, lipidic fracrion, 193, 205
Tuberculin, 208
Tuberculosis.203
Tularemia, 197
Tumors, see also cancer, different agents
- a n d c a l c i u m ,3 5 9
- a n d c h l o r i d ec o n t e D t ,3 1 2 , 7 1 0 , 7 1 6
- and cholesterol.370
- and conjugatedfatty acids,322
1 a
I
I t l
I N D E X
'I'umors
S t i tn u t r
a n d 1 1 l . r l . l 1 c \ 1 1 1 .I < i
and nr:rng;rncs*, 1(ii)
massi!c,
a n d n t r r c i l F l i r n \ . . 1l l . " l l ?
f l n d r l l r t l t \ ' l i l ) { , l . r n l l l r en u , ! 0 { ) . l l . 1
and rrrlllirdr:nulrr_ i!)9
mullinlr.:10
n e c f ( ) l l \ ' l r t t i . lt t c r r r . 1 1 1 , r 1 1 r ll jf.,,{ }
ilnd ()l I f;rrlr 'i('t'l\. ilh. : I (l
__ pilpiilrrrrrrL. :!ll
-* and
l r t r l i r s r r i r n t .1 ' l f r
*and
r : r r l r l r l i o r tl .l , l , l r ' r 1 , 5 1 1 , ( l t i , : . 1 ( r
and rlctrrl." 1"1)i, | {rt. .iiV,.i7.i. .l(r/r.
I ' \
*and
. ' \ l { , . 1 - 1 6 .' 1 : : . J f i l . l ( t l
",,1;111.
* $Cf iilrrr .pt'r'ili. irrr'rtl'.
- a n d r u l l r . r rl l r r r ' t . , t , | .i J r ,
-UlCCr:ltrrl. ili
ti{'i
* A n d l t t t s t r p r r i t r l t . rltt',l r( . l r , r r t r l, l l r i i - * 1
46li
* i f n d r j r u . r . r . : ( ) t t .i ( i i
- W a l l c r ' r . : 0 9 , , lj 6 . - { l ) l. r r l } N ,{ ) l J , r ' i I
- and /rni, -.1()l
Tung orl. i1{}
T u r p e n t i n c t r i l . i - 1i
T w i n f o r n t ; i t i r r n ,l l l . { r l l
* - i r r c i r r ei r r o t er r r . l i l
- - n r { t h } l e h d l ; i n t h r c n E 'l.l J
T y p e s t t f r r l l h t r l u r r r r r ' , ' e cl f i l . i i l ; r r r c r . \
rirrJ I)
- - g n p gj y l i ) l r l g i ( c l f ej t . J r l { l
T y p h o i d ,1 9 7 . : { } 1 . : l n
Typhus, lql
iir hnrirrrlrop ir proi:r:rr. l l l
Ulcrnlilor
- a n d l i p i r l s . l { r { 1 .3 3 . 1 ,i , 5 l
JhX
r r I : . 1 . .t r r . . - l ' , ,
. i l i , i i c . $ r ' r t t '"l 1 1 1 1 r 1/ 1r i. X
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;rtl!l \lt(i(k.
I ll.
6ti()
r r r r r .t ll l r l r . p l i r r r t r I. I { }
- . - t i l t i ! r i r r r r r . r .l 9 l
[ : r r r u l r t l l r l c d1 1 1 ' J 1 | 1 r !] t; u1 t11' 1 -iyt . tr ; r n c i d i l ! .
r:q
urN.rnr.
. r r l i l f ; l l n . . . 1l '
, ' l 1 ' l . r . r '' t. .
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rr,i,
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L i r l l - ' l t l L l , : , i . . l . r l , r : ,l i { r
, l l t ' t l , , t l . . . r . ! \. i r i \ : l t i , r l ] t | l ! r
| \ ! r r t l t L ' t t , r n r l f a t c J ) 1 1 ( r tr tn t l c f , 5 l ]
i:lrrtttrrrtti;
.i.i,l.
o.l I
1 1 ( l lf n 1 t i l l .
l.i(r.
,119.
5 r l (I
rt,,rtltrtttg \lttr\lilncc\. \r.C pcroxrdct
Ir,ltlfrn\.9{}
f ) ( t r r \ l ( l r : \ . l i ( ) . l 1 - 1. - ' i l .
f r Xi
. 1t ' ) . 5 : J .
I t r r t c I l l , J . l . ' J l ] l ( r l i . \ 1 1 . - i , 51
, t r t , . il ' i o i , d I i l r i l l ( 1 r i i a l L r r l i n i t l ' , - { 5 J
'
\ f \ t ' , : l l l fg r , r r i t t . X f ' ^ J 1 5 . 5 l :
... -.,irlllirtlt.\1.lfr6. .ral. i95
t rin,rli
-
\urtiicr lertrii'rrt"l{',
i l"l. :.1f)
1fi:,
; i r l { l i r r t tc t t r i c u l { ) t ] ] \ ' , . q B J (
(N.J
ttl ]tlittt:rls.
- . ; r t l t l L i r l l f( ' r i l r c r i p r , 5 1 1
'
, r l l r i . , - r l l i i t r l r{ ' ) { } , { } J H
l5l.
772
INDEX
Virus pathogencity
- place in organization,26, 690
- plural activity, 293
- and seasonalchanges,297
- and tumors. 290-301
- two types, 290
V i t a m i n sA , B ' , B r , 8 " , D , E . K a n d S . d . c .
pH, 604
Volcanoes, 108
WurEn tumor, 326
--and
lipids,625
- - a n d S . d . c .p H , 6 0 8
- - variation, 653
-and zinc,401
W a t e r c i r c u l a t i o na n d o r g a n i z a t i o n .3 0
- e x c r e t i o n .1 6 6
- a n d n i t r o g e nm e t a b o l i s m ,5 9 5
W h e a l r e s o r p t i o n ,8 3 , l l 9
- - i n a b n o r m a l c o n d i t i o n .5 6 6
- - a n d o l d a g e ,4 1 9
Widows and cancer, 30E
Wounds, see also S.d.c. pH
- atonic and oxygen, 406
-and cblorides,2ll
- h e a l i n g ,l 6 l , 6 5 8
X-n.rys, see radiation
Y o u r n c h a r a c t e r s ,3 5 7 , 6 5 4
- - effect of lipids, 165
- - persistence,82, 66 I
Y o u t h a n d v i r u s e s ,1 5 6 . 6 5 4
Zrxc. 359. 401
- a n d c a l c i u m ,3 5 6
- and copper, 359
- in periodic chart, 105, 394
- p h a r m a c o l o g y .4 0 I
- and testesteratomas.298
- and various effects.401