CHAPTER
LIPIDS AND LIPOIDS
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 major importance in physiopathologicalmanifestationswith long-lasting
patterns.The study of the lipids has substantiatedthis.
However, before discussingthese substancesand their properties, a
nosologicalproblem must be considered:What are lipids? How can they
be defined?
DEFINITION
OF LIPIDS
The literature fails to furnish an adequatedefinition for the group of
that show thosepropertieswhich biochemistryand experimental
substances
biology attributeto the lipids, A definitionon a chemicalbasis,such as one
which considerslipids to be fatty acids and fatty acid derivatives,appears
to be insufficient. It excludes substancessuch as those forming insaponifiable fractions which not only have properties attributed to lipids, but
continuouslyintervenein the processesrelated to them.
such as "greasiness"and solubility come nearer
Physicalcharacteristics
to the real situation without providing a satisfactorydefinition. Bloor's
definition (Note /J, widely acccptedtoday in spite of having been found
inadequate,has introduced-in addition to the important solubility characteristics-certain less acceptable criteria such as the origin of these
substances
and the direct relationshipto fatty acids.Without thesecriteria,
lipids would have to include the group of hydrocarbonswhich have the
same solubility property but usually are not encounteredin organisms.
However, with these criteria, Bloor's definition, besideslimiting the field
too much through the requirementfor a relationshipto fatty acids, excludes the entire important group of synthetic agents with similar properties.To be complete,a definition would have to include these artificial
substances.
Thus confronted by the need for a satisfactorygeneral definition, we
proposedone in 1940 (23) which has since been of great help to us in
all our research: a Iipoid is a polar-nonpolar substancein which the nonpolar part is predominant. It is thus lormed by one or more polar groups
bound to one or more nonpolar groups, the last being energeticallypredominant. In terms of intervening forces, this definition considers the
cohesionforces of the nonpolar part, and especiallythose related to its
surfaceand known as the constant"b" of van der waals forces, which in
lipoids are predominant upon the electrostaticforces of the polar part.
The definitionhas provided the key for the study of the multiple problems
in which these substancesappear to be involved. This definition appears
acceptablesinceit explainsall the known prop€rtiesof the lipids. Further-
LTPTDSAND
LTPOTDS
/
ll5
more,the study of the specificrelationshipbetweenthe forces involved
couldevenpredictnew properties,
as will be shownlater.
The distinctionbetweennaturaland syntheticsubstances
as a basisfor
a definifioahasbeenobsoletein biochemistryfor a long time. In our study
however,
it appearedto be didacticallyusefulto indicatewhetheror not
a substanceis encounterednaturally in the organism.Therefore,while
adheringto our generaldefinition,we haveemployedthe term "lipoids"
for the entiregroup of polar-nonpolarsubstances
with a predominance
of
thenonpoliupan, and have conservedthe term "lipids" to designatethe
naturallyoccurringmembers.With this separation,
we havealso avoided
Hydro
id
Borderl
ine
Lipoid
I
FN\\\
FI\\\\\\\\\
eot.rGroup
Group
Sl llon-Polar
Ftc.6lbis. Schematic representationof the predominant relationship of the polar
and nonpolar parts in hydroids, borderline substancesand lipoids.
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
Lipoids may be subdividedaccordingto differentcriteria.
l. According to the polar group.
A. Lipoids classifiedaccording to the nature ol their polar group.
( -COOH)
1. Lipo-carboxylic acids
(-SH)
2. Lipo-thiols
(_sofi)
3. Lipo-sulfonic acids
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.)
B. Lipoids classified according to the predominantelement ol the polar
group.
l. Lipo-sulfur compounds
a. Lipo'thiols
b. Lipo-sulfonicacids
c. Lipo-sulfides
d. Lipo-sulfoxides
e. Lipo-sulfones
f. Lipo-sulfites
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,)
C. Lipoids classified according to the energetic character ol their polar
8roup.
A. Lipoids with negativepolar groups
l. Lipoacids
a. Lipo-carboxylicacids
b. Lipo-thiols
c. Lipo-sulfonicacids,etc.
2. Lipo'atdehydes
B. Lipoids with positivepolar groups
3. Lipobases
a. Lipo-amines
b. Lipo,guanidines
c. Lipo-imines,etc.
4. Lipo-alcohols
ll. According to the nonpolar group.
A. Lipoids classifiedaccording to the structure ol their hydrocarbon chain.
l. Aliphatic
2. Alicyclic
3. Aromatic
4. Heterocyclic,etc,
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
Physical Lipoidic Properties
Solubility
Solubility representsthe first and most important of thcselipoidic prop
erties.Characteristically,a lipoid has a greatersolubility in neutral solvents
than in water.This is explainedby the fact that the two constituentgroups,
polar and nonpolar,inducediffercntsolubilitypropertics,
to a frec movementbetwcen
As is well known, solubilitycorresponds
moleculesof the solventand the solute.(25) Solubilityis greaterwhen
the physicalpropertiesof the groupsformingthe solventand thoseforming
Lhesolutearc similar;it is impairedwhcn thcy are different.Consequently,
polar groupsin a solutewill tend to favor solubilityin solvcntswith polar
groups, such as water. At the same time, thcy will oppose solubility in
neutral solventsformed by nonpolargroups.On thc othcr hand, nonpolar
groupsin a substancewill favor solubilityin nonpolarneutralsolventsbut
will opposeit in polar solventssuch as watcr. Polar groups thus are hydrophilic and lipophobic,while nonpolarare lipophilicand hydrophobic.(26)
While the solubility characteristics
of substances
composedonly of
polar or nonpolargroupsare rcadily apparent,thc problcm is more complex when a substancecontainsboth polar and nonpolar groups. Since
groupswith antagonistic
such a compoundpossesses
solubilitytendencies,
"in
toto" will dcpendupon thc relationshipbctweenthe opits solubility
posing forces. For a borderline group of polar-nonpolarsubstanceswith
approximatelyequalforces,therc will be cqual solubilityin polar and nonpolar solvents.For other substances,
thc predonrinancc
of one or the other
group will determinesolubilitycharacteristics.
If thc electricalforces of
the polar group predominatc,the substanccwill be hydrosolublebut insolubleor only partly solublcin nonpolarsolvent.s.
If, on thc contrary,the
cohesion-i.e., the van der Waals forccs-of the nonpolar group predominate, the substancewill be solublein nonpolarsolventsand less,or even
not at all, solublein water. (Tnnn VI)
Polar and nonpolarforcescan be calculatedand thcir study can indicate the place of a substancein this systematization.
The importanceof
solubility for defining and systematizing
lipoids became apparent in a
physicomathematical
study of thescsubstances
carried out by Jean Mariani
in our laboratories.(Note 2) We havc defined as "hydroids" those substanceswith predominant polar groups which are morc soluble in polar
with no predominance
solventssuch as water.The "borderlinesubstances,"
of either group, show the same solubilityin polar and neutral solvents.
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
6
3 . 7( a t 1 6 " )
0.4
o.24
@
0 . 2 5( a t 1 0 0 " )
€
@
6
' 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
natein their molecules.
Therefore,they are not lipoids.n-Butyl alcohol,
moresolublein neutralsolventthan in water,thus is the first lipoidic member of this homologousseries.However,this is not true for all its isomers.
The primary,secondaryand iso butanolare the first in their respective
seriesto possessthe solubility propertiescharacteristicof lipoids. In the
tertiaryalcoholseries,however,the four+arbonmember,the tert.-butanol,
doesnot showthe samesolubilityproperties.Tert.-butanolis misciblewith
water and neutralsolventand as such,is not a tipoid. For this tertiary
alcoholseries,it is the five-carbonmember,the tert.-amylalcohol,which
first showsthe solubilitypropertiesof a lipoid,being only 12.57osoluble
in waterandinfinitelysolublein petroleum
ether.Thus,of the four isomers
of butyl alcohol,three are lipoids,while one, tert.-butylalcohol,is not.
( Trn l n V III)
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
@
€
6
@
J
€
t
J
a
@
A
4
4
4
5
5
5
5
5
5
6
6
6
8
r Names approved by International Union of Chemistry.
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
The same methods were used to recognizethe first Iipoidic membcrs
of various alkane derivativesstudied.-fnnle IX shows the first lipoid
membersof severalhomologousseries.
T,rgls IX
Flnsr Lrpololc MevneRs lN Vrnrous AlxeNr' Denrv,rrrve
Hovor-ocous SE'Rrr.s
S u b s t a n c eI
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.
Molecular l^oyer Formation
Other fundamentalcharacteristics
result from the different solubility
propertiesof the two parts, polar and nonpolar, forming a lipoid. Introducedin a diphasicmediumin which one phaseis watcr and the other oil
or even air, the polar group, with the tendencyto be solublcin water, will
penetratethe water. Since the nonpolar group, which is hydrophobic, is
predominant,not only will it not cnter the water but it will also prevent
theentire moleculefrom moving frecly in water.Conscquently,
the lipoid
moleculewill remain at the surfaceof the water with only its polar group
penetrating.
Becauseof this, the moleculewill assumean oricnted position
towardthe surfaceof water. If the secondphaseis a neutral solvent,the
lipoidmolecules,which will accumulateat the interphasewith the polar
group in water, will have the nonpolar group penetrating the ncutral
solvent.
In both cases,the moleculesform oricntedmolccurarlayerswhich, if
presentat the limit betweentwo phases,would appearas organizedformations.This property which appearsas a direct consequence
of the characteristic
constitutionof the lipoids has further consequcnces.
In such a
layer,the polar groups penetratingin water will influencethe properties
of its surface,and thus reduceits surfacetension.
124 /
REsEARcHrN pHystopATHoLocy
Through the coulombiancharacterof their electrostaticforces,the polar
groups will thus confer, accordingto their nature. a positive or negative
electricalcharacterto the layer.
In a mixture of water and oil, the presenceof a lipoid layer will lower
the intersurfacetension and will favor the breaking down of the phases,
facilitatingthe formation of an emulsion.The presenceof the same electrical chargeat the surfaceof theseresultingemulsiondroplets will act as
a repeUentforce betweenthem and increasethe stability of the emulsion.
(FiS.62) This is anotherimportantcharacteristic
of lipoids which results
from their peculiar constitution.
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
LtPtDS AND LtPOtDs
/
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
their insolubility in water, the orientation of lipoids has allowed them to
play a very important role in biology.
The very existence of biological entities appears to depend upon the
ability of lipids to build up boundary formations separating and thus assuring the individuality of biological entities.
Through peculiar, reciprocallyopposedorientations,two or more layers
of lipids can form a membrane with two polar faces which has the ability
to separatetwo aqueousmedia. In its simplestform, such a membrane
appears in mitochondria. (Note 3/ Similar boundary formations identify
nuclei and cells and appear in higher entities, as in the membranes and
intercellular cements of lymphatic and blood vessel endothelia. It is this
peculiar orientation which allows lipids to establishthe necessaryboundary
formations resulting in complex hierarchic organisms.The existenceof
biological entities, at least from the chromosome level up (and probably
even below that level), can be seento result directly from the intervention
of lipids as a separatesystem,particularlyin the formation of the dipolar
lipidic boundaries.
However, boundary formations which separatethe biological entities
would not have been efficient if they did not fulfill another capital role:
that of allowing selective passageof metabolites. A totally impermeable
membranewould isolate the respectiveentities and result in their death.
On the other hand, a totally permeablemembranewould have no usefulness.The boundaryformation has to act selectively,pcrmitting the passage
of some,but not all. substances.
But even this does not seemto be sufficient to insure an efficientboundary. Most important, such a membrane
must be able to alter its permeability,quantitativelyand qualitatively,according to variationsin circumstances.
Such capacityfor altering perrneability can be relatedto the presenceof the two groups of lipids, fafty acids
and sterols,with their antagonisticpropertiesrelating to permeability.
The fatty acids appear to induce permeabilityin the membranethey
form, especiallypermeabilityfor anions.The perpendicularposition to the
surfaceof water assumedby the nonpolar aliphatic groups when the fatty
acids form this boundary membraneappearsto be favorable for the passageof a substancethrough the membrane.The fatty acid moleculesthus
can be separated,permitting other moleculesto pass between them; that
is, to passthrough the membraneformed by the fatty acids.The negative
electricalcharacterof the polar groups of these fatty acids explainswhy
they representa kind of barrier to the free passageof cations.Thesecations
are attractedand retainedby the acid polar group. This would explain the
manifestchangesin permeabilityunder the influenceof calcium ion. The
LtPTDS AND
LTPOTDS
/
127
removalo[ calciumfrom cellularmembranes,
throughtrcatmentwith oxalates, increasespermeability,while trcatnrentwith calcium salts reduces
permeability.The bivalentcalcium ion, when it binds the polar groups
of two adjacentfatty acid moleculcsin the membrane,prcventsthe passage
of other moleculesbetweenthcse parts o[ thc membrane,a fact which
explainsthe manifestdecrease
in permeabilityinducedby this cation.
The other group of lipids, the sterols,have an effect on permcability
opposite to that of fatty acids. This can be related in part to the bond
rvhich these sterols make with the fatty acids. Consequently,they block
any passagethrough the part of the membraneformed by fatty acids.The
impermeabilityis due, to someextcnt, to a peculiarityof the layersformed
by thesesterolsthemselves,
The polycyclicmolcculesof sterolsdo not take
the same perpendicularposition toward the surfaceof water as fatty acids
d o . ( 2 7 ) ( F i S . 6 J l S i n c es t e r o lm o l e c u l eass s u m ea p o s i t i o na l m o s tp a r a l l e l
(a)
-ll+llllll
ooooooooc'
Fattyacidorientedlayr
ratgr
(b)
o
/z///,///,.
.
rater
oooooooo
Sterol orientedlayer
l.-
polar group
rrcn-polargroup
Ftc. 63. Schenraticuspcct ol orientcd interluce lu1'ers.The perpendicularposition to
t h e s u r f a c eo f w a t e r o f f a t t y a c i d m o l e c u l e s( a ) f a v o r s t h e p a s s a g eo f o t h e r m o l e c u l e s
t h r o u g h t h e s e p a r a t i n gm e m b r a n e st h e y f o r m . O p p o s i t e l y t, h e a l m o s t p a r a l l e l p o s i t i o n
t o t h e s u r f a c eo f w a t e r o f t h e p o l y c y c l i cm o l e c u l c so f s t c r o l s ( b ) p r e v e n t st h e p a s s a g e o f o t h e r m o l e c u l e st h r o u g h t h e m e m b r a n et h e y f o r m ,
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
the energeticcenterpresent.In one which occursat room temperatureonly
for the conjugatedfatty acids,such as eleostearicacid, and at 100"c only
to some extent for oleic, linoleic and linolenic acid, the oxidation leads to
thc appearance
of peroxides.(29) In anotherform of this oxidation,taking
place for oleic. linoleic and linolenicfatty acids at room temperatureor
below 50"c, hydroperoxidesresult, as it has been shown by Farmer and
corvorkers,
first for rubber (30) and afterfor fats (31). Another important
fact seen in rancidity changesis that the atmosphcricoxidation of polyethenoidfatty acidscan resultin a displacement
of the double bondswith
the appearance
of conjugatedisomcrs.(32)
The study of natural rancidityhas represented
the basicguide for our
study and systematization
of the processes
encounteredin normal and abnormal physiology. we searchedand found this similarity not only in
generaloutlines,but also for most of their details.By referringto the processesfound in rancidity, we were able to identify, besidesenzymaticlipolysis and enzymaticKnoop beta oxidation,known to occur in the organism,
also the interventionof hydroperoxides,peroxidesand the conjugationof
double bonds. Not only the processesthemselvesbut a.lsothe conditions
under which they take place and their inter-relationship
have been found
p
a
r
a
l
l
e
l
to
i n v i v o t h o s ew h i c h c a n b e s e e ni n v i t r o .
We rvill seeall along in the studyof fatty acidshow far the biological
interventionof this parallelismgoes.
Caloric Metabolism
Although all the fatty acidsarc ulrimatelyusedby differentorganisms
as caloric metabolites,the saturatedand monocthenicmembersare most
importantfrom this point of view. Among the saturatedand monoethenic
fatty acids,the memberswith long chainsappcar to be thosc which are
kept in reservefor caloric purposes.we could show that the principal form
of caloric desmolysis,
the Knoop beta oxidarion,takes place directly, almost exclusively,on memberswith relativelyshort chains,tlat is, with a
maximumof l0 or 72 carbons.
While fatty acids with short chainstake part directly in thesecaloric
metabolicchanges,thosewith longercarbonchainsmust undergopreliminary changesbefore entering into caloric metabolism.A desaturation,
changinga saturatedfatty acid into a monoethenic,appearsto be a first
stepin caloricmetabolismof the long chain members.The monoethenoids
thus can be seento be intermediaryforms bctwcenthe saturatedreserve
and the short-chain,easilymetabolized
fatty acids.
The double bond in these monoethenicacids would rhus appear to
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
such a process,the oxygen is libcratedas a free radical, the entire process
thus corresponding
to an activationof oxygen.The changeof a molecular
oxygcn into a frec radical would representthe physiologicalrole of unsaturatcdfatty acidsin oxidationprocesses.
The presenceof two double bondsin non-parallelposition,contmon to
most of the naturallyoccurringpolyunsaturatcd
fatty acids,is even morc
important; the two double bonds exert a parlicularly strong influcnce on
the specialcarbonwhich is in thc intermediarypositionbetweenthem. Becauseof the alternateinduction producedby the strongJypositive carbon
of the carboxyl,the carbonsof the chainhave alternatecharacters,
positive
and negative.When an intermediarycarbonalso has a strongpositivecharacter, it appcarsto be cspeciallyable to fix oxygen.This strong.lypositive
intermediarycarbon,occurringin naturalpolyunsaturated
fatty acidswith
more than two double bonds, may be the rcason for the important role
playedby theseacids whcn they act as essentialfatty acids in the organism. (Note 7)
The study of rancidity has further shown that, while the in vitro
oxidationof an unsaturated
fatty acid under mild conditionssuch as room
temperatureleads to the appearanceof hydroperoxidcs,oxidation at a
higher temperaturewill result in anothcrfixation of oxygen,this time at
the doublebond itself.Epoxidcsor peroxidcswill appearaccordingto the
ionic or molecularcharactcr of the oxygen. This extremely important
processalso occurs in rancidity under the inl'luenceof an enzyme. It is
highly probablc that a similar proccsstakes placc in vivo in those pathologica.l
conditionsin which pcroxidesappearin the urine.Radiation,certain
inflammations(especiallythosc due to streptococci),administrationof
seleniumpreparations
or of highlypolyunsaturated
fatty acidsare followed
by the appearanceof theseoxidizingsubstances
in urine. As mentioned
above, when these substancesappear, there also arc increasesin indoxyl
and glucuronicacid, which can be considered,up to a certain point, to
resultfrom abnormallyintensiveoxidationtakingplaccon tryptophaneand
glucose.(See below.) while acrivationof oxygenis a physiologicalprocess,peroxidesappearunder abnormalconditions.
ABNORN{AL FATTY ACIDS
T h e s t u d yo f t h e r e l a t i o n s h i bp e t r v c c na b n o r m a cl o n d i t i o n sa n d l i p i d s h a s
progrcssively
led us to considerthe existence
of clualitative
changesin these
l i p i d s ,b e s i d e st h e c l u a n t i t a t i voen e s .T h e c x i s t c n c eo f a b n o r m a ln r e t a b o l i c
processes,
and espcciallythe fact that such abnormalitiesare often of long
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
Methods ol Investigation Used
Followingthis line,we firstinvestigated
the forms underwhich the lipids
in gcneralare presentin the organism.We utilizedthe differences
in solubility betweenthesedifferentforms, separatingthem into free lipids, lipids
kept in a labile bond with other constituents,
as in cenapse,lipids bound
throughtheir polar group as in fats,or in the still strongerform as lipids in
combinationsso firm that thcy cannot be separatedexcept through saponification.The methoddevisedfor this study and someexamplesare in Note
8A. This researchshowedthat under abnormalconditions,very important
variationsoccur in the amountsof the difterent forms. This study pointed
out that the free lipids are greatlyresponsible
for the importantnranifestations in which lipids appearas activeagents.
In the study concerningthe abnormal metabolismof the carboxyl and
nearby carbons, we investigatedthe appearanceof fatty aldehydesor
ketonesin blood, urine and in the cells.
Onc of the major problemsencountered
was the appearance
in vivo o[
conjugatedfatty acids,as abnormalfatty acids.In order to ascertaintheir
presenceand to measuretheir amounts,we had utilized three differcnt
methodsof investigation:spectralanalysisin ultra-violetand in the first
portion of the visible spectrum(Note 8Bl; the study of the place of thc
doublebond in the fatty acidsmoleculethroughthe fissionof thesemoleculesand the analysesof the resultingfractions.(SeeNote I, Chapter 101
More recentlywe have tried the vapor fractionationmethod (gas chromatography) (Note 8C).
The first, and especiallythe secondmethod,gave us valuabledata permitting us to recognizethe interventionof conjugatedfatty acids in ab'l'hese
normal conditions.
studiesrevealedthe appearanceof conjugated
fatty acids,especiallyas trienes,the increaseof their amountwith the progressof the conditionsand especiallythe fact that deathoccurswhen their
concentration
in the bodieshasreacheda criticalvalue.This has markedthe
importanceof thesesubstances
in physiopathology.
The fact that gaschromatographydid not revealthe presence
of conjugatedfatty acidsappearsdue
to the conditionsunder which the methodactuallyworks.
Later, we will frequcntlyreturn to the variousproblemsrelatedto interventionof conjugatedfatty acids.We could thus directly correlatethe
interventionof theseabnormalfatty acidswith the pathogenesis
of the manifestationsof many conditionssucb as trauma,shock,adrenalectomy,
and
especiallywith the noxiousmanifestations
following irradiation.(Chapter
I0)
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
Gl;'cerol and Glyc'erophosphoricAcid
It is classically acceptedthat the intestinal absorption and circulation
of fatty acids is made through bonding to varioussubstances.
The analysis
of this absorption shows, however,that different fatty acids have preferential bonds. For saturatedand mono€thenicfatty acids,the bond is principally with glycerol. Although mono- and di-glyceridescan be identified
in the cells of the intestinalmucosa,these fatty acids leave the intestine
as triglycerides,forming the largest part of the chylomicrons.They are
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
rc'ductionof the fatty acid's activity, aflectingoxidative processesdirectly,
or indirectly through reductionof membraneFrcrmeability.
while one group of anti-fatty acidscan be directly relatedto hydroids,
and especiallyto glycerolor to glycerolbound to phosphoricacid as in the
ion, a secondgroup is represented
by lipoids, principally
-elycerophosphoric
formed by derivativesof a characteristicring system,the cyclopentanophenanthrene.As anti-fatty acid lipoids, thesc compounds,the steroids,
were of special intcrest in lipoid research.only some aspectsof the biologicalactivity of steroids-mainly, thosewhich reprcsentnew views in the
studyof thesesubstances-willbe discussed
here.
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
steroidshas permitted us to recognizethe conditions which induce stronger
activity for these polar groups, conditions which are usually fulfilled in the
naturally occurringmembers.It could thus be seenthat the reactivityof an
oxygen bound to Cs is increasedif another double bond present in the
cycle is parallel to the double bond through which the oxygen is bound to
Cr.A doublebond betweenC1 and Cr,,as shownin Figure 64 (a), fulfills
(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 ) .
such a condition.A similar influenceis exertedindirectly by a double bond
betweenC6 zrndCz (b) which, through induction,will influencethe parallel
Cr and C5 bond and further the double bond of the oxygen.This explains
the influenceexertedby the double bond presentbetweenC, and C, (c).
as in the synthetic,prednisolone.
Further enhancement
of reactivitywould
be obtainedwith a third doublebond addedbetweenC6 and C7. The parallelism bctwecn three double bonds (d ) would produce an increased
reactivity.
For the hydroxyl, a similar enhancedreactivity is induced by double
bonding of the carbon to which the hydroxyl is attachedwith a double
bond for the Cs - Ca or C, - C., as shown in Figure 65 (a and b). A
similar condition is fulfillcd if a doublc bond is prescntin the moleculc
parallelto any of thesebonds,as seenin Figure 65 (c) where the double
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
bond is betweenC5 and Co. An enhancedreactivityof thesecompounds
would be obtainedwith one double bond betweenC3 and C. and another
betweenCr and C,, (d ).
We will come back to this important intervention of the double bonds
in cyclic molecules.
The energeticproperty of the cyclopentanegroup app€arsto be correlated with its odd number of carbons.The a.lternatesuccessionof carbons
with positive and negativecharactersresultingfrom the induction effect
causestwo carbonsof this cycle to have the sarnesign. This "twin formation" inducesa specialmolccular reactivity related to the pentanic cycle
of the steroidmolecule.(Note 9)
The special reactivity seen for c3 of thc cyclopentanophcnanthrene
molecule can be explained through a hypothesiscovering the origin of
these substances.
Although the origin of a cholesterolmoleculethrough a
cyclizationof squalene(35) appearsplausible,this seemsless probable
for the corticoids.We havetried to connecttheir origin to arachidonicacid.
Severalconsiderationssuch as the high levelsof arachidonicacid and
corticoids in the adrcnals,and the reduction of the former when an important amount of the latter is excreted(Note I0), seem to establisha
correlation between these substances.According to our hypothesis,the
steroidswith a two carbon chain at Cr;, as seenpresentin the corticoids
and luteoids,would result from a cyclization of the arachidonicmolecule.
(Note 11l This would explain the specialreactivityof C., which woutd
correspond to ce of the arachidonicacid bound in this molecule by a
double bond.
A study of the different steroidsunder this energeticaspecthas permitted us to understandtheir physiologicpropcrties.
with the c3 having a hydroxyl or an oxygen as polar group in almost
all the steroids,the variety of the biologicalproperrieswould be relatedto
the different conditionsat the other extremity of the molecule,principally
at Crz, which result from the specia.lenergeticconditionsprevalentat this
regionof the molecule.The simpleststeroidsare thosehavinga polar group
representedby an OH or O fixcd at C17.Such naturally occurring steroids
have propertiesrelated to secondarysex characteristics.We will discuss
them briefly here.
Sex flormones
This group of steroidshas two polar groups, one at C.qand one at C17.
The energeticcenter at C3 can have negativeor positive polar characters,
accordingto the presenceof oxygen or hydroxyl. The energeticcenter at
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
and the hydroxyl at C3, the reverseof testosterone.In both substances,
testosteroneand androsterone,the two conditions for testoid activity, positive-negative polarity and the same relative position between the polar
groups, are fulfi.lled.(Fis. 67)The differenceswhich exist betweenthese
substancesin their specifichormonal activity can be explained through the
difterent influence exerted upon the two polar groups in these substances
by the rest of the molecules.
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
the center at Cg has a nucleophilic character. This is reinforced by the
presenceof a double bond botweenC5 and C6 which, by paralleling the
bond betweenC3 and C1, increasesits ionic character and consequently
the reactivity of the hydroxyl bond to C3. Through this hydroxyl, sterols
combine in general with substanceshaving a negative polar group to form
esters.
Besidesthe capacityto combine with fatty acids in general,one of the
most important characteristicsof the principal sterol of animals, cholesterol, is its selectiveaffinity for certainfatty acid membcrs,the polyunsaturated.We tried to explain the specificityof this bond through an interesting
processwhich could be called "steric coupling."
In this process,two molecules,usually lipoids, are kept together not
only by the combination of their polar groups but also through a bond
betweentheir nonpolar parts. The two moleculesare reciprocallyattracted
through the multiple forces presentin the nonpolar groups. Some are related to attached centers, while some, such as those correspondingto
cohesionforces,are relatedto the rings themselves.An important factor is
the rigidiry of the sterol moleculewhich permits another molecule,if it is
flexible,to make the steric coupling.The rigid skelctonnot only kecpsthe
energeticcentersof one moleculein a fixed positionbut permits the flexible
aliphatic chain to cover over the polycyclic molecule and thus bring the
energeticcenters of one molecule in contact with those of the other.
Through this, steric coupling completesthe bonding of the polar groups.
The greaterthe concordanccbetwcencnergcticcentcrsin both molecules,
the more perfect the coupling is, for the more complete is the reciprocal
neutralizationof the energeticcentersof the two molecules.Stericcoupling
explains why, of all the fatfy acids presentin the organism, cholesterol
seemsto prefer to bind thosewith polyunsaturatedchains.It is thesefatty
acidswhich have severalenergeticcentersin thc nonpolar group as representedby double bonds. The long chains of these fatty acid molecules,
havinga certaindegreeof flexibility,will then completcthe stericcoupling.
(Note I2 )
Stericcoupling, in addition to its generalimportancein biology, where
it representsa kind of molecular reactivity,seemsto cxplain the antagonistic influence exercisedby different constituents,especiallythe sterols
and polyunsaturatedfatty acids.Through stericcoupling.cholesterolcould
influencethe activity of thesefatty acids more dircctly related to the nonpolar group. It has to be emphasized,however, that the neutralization
resultingfrom steric coupling is not irreversible,On thc contrary, through
the interventionof various factors,such as the breakingdown of the bond
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.
Steroids with a Two-Carbon Chain
Among the most important steroids are those having a two-carbon chain
fixed at C17.
Two groups, the Iuteoids and corticoids, appear directly related to
allopregnanehydrocarbon,the steroidpolycyclewith a two-carbonlateral
chain fixed at Cr;. As we have alreadyseenin the hypothesisconcerning
the origin of the steroids(Note 11l, this hydrocarboncould have been directly derived from arachidonicacid, thc two-carbonlateral chain correspondingto the tail chain of this acid, a tail which remainsafter cvclization.
The Luteoids
The prototype of the luteoids is progesterone.Two polar groups C : O
are present,one at C3 of the polycycleand the other at C:roof thc tail chain.
A paralleldouble bond betweenCa and C; completesthe formula. Energetically,progesteronepresentsa first center at C3 which appearsstrongly
nucleophilicfor two reasons:first, becauseit correspondsto the potent
electronegative
Ca and second,becauseit is reinforcedby a double bond
presentbetweenC1 and C5,and whichis henceparallelwith the doublebond
of the carbonyl.The second : O is attachedto the Cs,yof thc tail chain.
This alsoappearsreinforced,the doublebond of this carbonylbeingparatlel
to the bond betweenthe Crr and C17,which in the cyclopentane,
accordhypothesis
of
ing to the
twin carbons,binds two negativechargedcarbons.
Through its constitutionprogcsterone
is also a lipoid, thc complexhydrocarbon group being predominant over the polar groups. With its polar
nucleophilicccnters,progesterone
has the fundamentalcharacterof acid
luteoid activitycorresponds
lipoids. Progesterone's
to the presenceof two
relativelystrong neutrophiliccenterskept in the characteristicpositions,
one at C3 and the other at C2s.
We can see that any disturbancein this energeticpicture, any change
from the dinucleophilicat any center, decreasesthe luteoid propertiesof
the substance.With more profound changes,the luteoid activity is even
(Note 13)
suppressed.
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
syntheses.[n difterent corticoids the constitution of the Crr : Czr formation
will differ and this will determine which substanceis to be synthesizedby
the respectivemold or templateformation.(Note I6)
Using the template hypothesis, we studied an entire series of body
constituentsforming the "gluco group." Glucose,galactose,glucosamine
and galactosamine,with their respectiveacids, as well as ascorbic acid,
:ue rmong thesessubstances.According to our hypothesis,thes€ neoglucogenic corticoids would have the important role of producing, possibly
along with other mechanisms,the entire seriesof "gluco" constituents.The
existence of difterent template formations would result in a variety of
synthesizedconstituents.
The intervention of the template formation in synthesis can occur
again and again without aftecting the molecule of the corticoid as such. It
is interesting to note here a structural curiosity which could be interpreted
as being related to template activity. In this template,the group of successivec,r, c,r, c13 and crz ?re part of the rigid skeletonof the cyclic
molecule,while C:o and Czr are forming the lateral chain attachedto C17.
This can be regarded as conferring a certain prop€r mobility to this lateral
chain as related to the polycycle.It is conceivablethat this lateral chain
would becomea closedformation when synthesistakes place.A movement
of the chain at Crz would permit the mold to open and thus liberate the
synthesizedmolecule. It is interesting to note here the importance of the
structure of the template for the constitution of the substancessynthesized.
Besidesthe polar group at c17, that at Crr is also important for neogluco
genic activity since it insuresa six-carbonchain in the synthesizedmolecules. A hydroxyl or carboxyl at the Co of the synthesizedsubstancewill
app€ar, according to the nature of the polar group at Cn of the steroid.
The respectivecharactersand positionsof Czr and C12 will permit thc
appearanceof a cycle formed by five carbons and an oxygen, characteristic
for the pyranic form of newly synthesized
substances.
An interesting confirmation of the template hypothesis was obtained
when glucosamine which, according to the hypothesis,is synthesizedby
the cortisonemolecule,was found to inducein patientsmany of the clinical
changeswhich are obtainedby treatmentwith cortisone.We will consider
these resultslater in our discussionof therapy.The capital role played by
glucosamine,galactosamineand the respectiveuronic acids in the constitution of the connective tissue representsthe "missing link" for the
explanation of the relationship betweencortisone,the other neoglucogenic
corticoids, and this tissue. Some part of the therapeutic eftect obtained
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
with theseneoglucogeniccorticoidsin diseasesof the connectivetissuehas
to be attributed to the intervention of the amino sugars.
In the study of anti-fatty acid activity, glycerol and glycerophosphoric
ion werefound to control the absorptionand circulationof saturatedmono-,
di-, or tri-unsaturatedfree fatty acids.The sterolsappearto counterbalance
the normal polyunsaturatedmemberswhile adrenal corticoids, and especially the neoglucogeniccorticoids, counteract the toxicity of fatty acids in
general and of the abnormal conjugated members in particular. Research
done in our laboratoriesby E. F. Taskier indicatesthat the adrenalsintervenein the defensemechanismagainstfatty acids,and especiallyagainst
the conjugated members which appear to be related to abnormal conditions. (Note 17)
The part of our researchconcernedwith the role of lipoids in normal
and abnormal physiologyhas been almost entirely guided by the concept
of an antagonismbetween the two groups, one with a positive and the
other with a negativepolar character.This specificaspecthas led us to
study, together with the fatty acids and the anti-fatty acids, other substancesrelated to this antagonism.In the group of Iipoacidsor acidic lip
oids, as obtained from tissues,organs or organisms,we rccognizedthe
group of porphyrinic acids,relatedto various hemespresentin the organisms. In the group of anti-fatty acids obtained from the same sources,
differentconstituentsform the insaponifiablefractions.
As related to this dualistic aspectwe have studicd another group of
substances,
which appearto act in the organismagainstthe anti-fatty acids
themselves.These other substanceswould representa kind of biological
brake to counteract an exaggeratedintervcntion of anti-fatty acid constituents,
we have made a specialinvestigationof two substancesof this group,
glucuronic and sulfuric acid anions, which charactcristicallyseem to opposecertain anti-fatty acid substances.
Thesesubstances
appearas a result
of an exaggerated
oxidationof normalmetabolites.
Under abnormalconditions, the oxygen resultingfrom the interventionof peroxidemay be fixed
to carbohydrateseven bcfore they have undergonethe preliminary fermentativetransformationsseenin normal metabolism.with the aldehyde
group bound to phosphoricacid, the oxidationtakesplaceat c6, the second
most reactivecarbon h the molecule.This direct oxidation would represent, accordingto our view, one of the sourcesof glucuronic acid. Similarly the sulfuric anion would result from the oxidation of sulfur present
in the organism.They correspondto the oxygenphascof offba.lance
D.
LIPIDS AND
LIPOIDS
/
147
Glucuronic and Sulfuric Anions
urine specimensthat contain abnormal oxidizing substancesshow significant amounts of glucuronic and sulfuric acid compounds.(Nore Ig)
The analysisof the conditions under which these two substancesexert
anti-toxic activity permits a better understandingof their role in general
biology. A certain parailelism exists, and has always been emphasized,
between a detoxifying and an etminating function exerted by these two
radicals. Not only do sulfo- and glucurono-derivativesappear in the urine,
but it often has been noted that glucuronic acid intervenes when large
amounts of certain substances,such as menthol and phenol, are present
and there are insufficient sulfuric acid radicals to insure detoxification and
elimination. when mineral sulfatesare administered,the proportion of
sulfederivatives increases.
This parellelism appearsespeciallyinterestingwhen we recognizethat
sulfuric acid representsthe end result of the oxidation of sulfur introduced
into the organism in combinations in which it is a bivalent negative element. Only a smaller amount of sulfur is introduced as a hexavalentpositive element: that is, as sulfate. Sulfur is introduced mostly in bivalent
negative form, as in methionine,cystine, etc. Both sulfuric and glucuronic
acid result from oxidative processes,acting in one case upon the thiol
group and in the other upon glucose.
The relationship botweensulfuric and glucuronic acid goes still further.
It has been noted that glucuronic acid appearswhen enough sulfuric radicals necessaryfor detoxifying action are not available. However, this is
not entirely true since one processdoes not duplicate the other. Qualitative
di.fferencesintervene. (Note I9)
The significanceof glucuronic acid in the defense mechanism seems
clearer when we recognizethat, with but few exceptions such as benzoic
acid, all the substanceswith which glucuronicacid combinesare lipids or
lipoids having one or more positive polar groups. The combination with
glucuronic acid takes place through these positive polar groups. (Note 20)
In our view, glucuronic acid like sulfuric acid, has a specific role in the
defense of the organism and this seemsto be directed especially against
lipids or lipoids with a positive polar group. Bound by glucuronic acids,
the latter are eliminated as excrementalsubstances.Glucuronic acid thus
would act against many anti-fatty acid agents.We can conceiveof sulfuric
and glucuronic acids as mears by which organisms are protected against
an exaggerated activity of anti-fatty acid agents. Along the same lines,
when lipoids with positive polar groups are predominant and able to act
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
tematizationof these variations would help us understandmany of the
processesencounteredin normal and abnormal physiology.
As we have mentioned before, the balance between two antagonistic
forces, especially for normal conditions, is not static. Instead, there is
alternating predominance of the forces, which results in an oscillatory
movement. Several groups of such coupled forces, each group with its
proper rhythm, are at work. Operating simultaneously,they make for a
series of very complicated variations. Yet analysis is possible since each
of the variations follows a dualistic pattern. The variations, as they occur
at different levels and with different intensities,have been identified
through various tests. In a secondstep they have been tentativelycorrelated with changesin lipids. And next, didactically,lipid changeshave
been related to various etiologic factors, some intrinsic and some extrinsic.
^Se.r
The influence exerted by the sex of the organism upon l-ipidicbalance
was brought to our attention by a curious effect seen when cholesterolwas
administeredin an ether-oil solution to rats. only the females showed
paraplegiaand ulcerationsof the hind legs.While castrationor administration of sex hormonesdid not alter this response,it was influencedby the
administrationof two groups of lipids. The insaponifiablefraction of human placenta,for instance,was seen to induce a high sensitivityto this
preparation of cholesterol even for males, while the acid lipidic fraction
of placentapreventedparaplegiain females.(Note 2l)
Similarly,the fact that in femalesalone,adipouscells appearedquickly
in the skin of the ear after the application of sulfur mustard could be
related to the interventionof the insaponifiablefractions. (Note 22)
Starting with these observations,it could be seen that, in generar,a
higher proportion of positive lipids exists in females than in males. This
could be shown by direct analysesand by analysesof manifestationsrelated to such lipids. while many differencesare to be seen in various
manifestationsbetween females and males, only some could be related
to the direct or indirect interventionof sex hormones.In such instances.
castrationwith or without the administrationof sex hormoneswas able to
change,and even to reverse,the differencesin manifestationsseenbetween
sexes.However, in instancesin which thesemeasureswere without effect,
the differencescould be relatedto rhe interventionof lipids.
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.
changescomparableto an offbalancetype D. This phaseis followed by a
much longer phase of type A, indicating sterol predominance.Intensive
rnentalexerciseproducesa markedchangesimilar to offbalanceA, with all
urinary testsshowingthe patternsfound with predominanccof sterols.
The responsesattributed to influencesexerted by external lactors could
be integrated in tle same dualistic mechanism,All the data indicate the
manifest influenceexerted by the time of day. Two marked changesare
seen,one around four o'clock in the morning and the other usually around
eight or nine o'clock in the evening.The morning changecorrespondsto a
predominanceof sterols,the eveningto predominanceof fatty acids.These
changes together with the clinical manilestations related to time of day
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
/
could be observedrelated to the atmospherichumidity. Other tests as well,
such as urinary pH, calcium excretion,etc., show a similar relationship but
to a much lesserdegree.(Note 28) The influence exerted by the environment could explain the changesseenfrom one day to the other in various
analyses.(Note 28)
t2
t,
q,
L
=
5t
.\t ao
(-
c,
o- 32
E
o,
6'
v,
(u
.U
z
v,
(a
s5
vt
(u
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E
(u
6t
l!
./,
q
i
9
b
r
t
z
!
,
t
6
t
0aYs
l
t
E
t
t
2
2
2
s
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.
ths influence exerted by changesin seasonswas studied. An increase
in fatty acids in winter and of sterols in summer could be noted. These
increa-sesalso could be largely related to the seasonalvariations in temperature. Very hot days were marked by analysesindicating intervention of
lipids with a positive character. The influence exerted by the seasonswas
s€en even in the responsesof organismsto pathological conditions such as
t.:t,t:i
154 /
nESEARcHtN pHysropATHoLocy
tumors; variations in character and growth of experimental cancer could
be noted. (Note 29) The relationship of many viral infectious diseasesto
seasonalchangeswhich has been noted in many epidemiologicalstudies
coufd be rclated to the changesin lipids. (Note 30)
Efiect ol Antagonistic Lipids a Difierent Levels
A more complete study of lipids under the dualistic concept was made
by considering their activity at different levels of hierarchic organization.
This researchwas greatly facilitated by the degree of individuality which
different biological levels exhibit when they are part of the hierarchic organizationof complex organisms.It was also aided by the availability of
lower organismsin nature which correspondto various hierarchic levels.
Through this double approach,the information obtained showed the importance of the relationshipbetweenthe levels of the complex organism
and the influence exerted by lipoids. If high doses of the agents are applied, the inJluenceis exertedupon all the levels.A preferentialinfluence
is exertedupon a singlelevel if reduceddosesare used.when medium size
dosesare administered,to the preferentialeffectupon one level a reactional
responseat other levelsis added.This resultsoften in concomitantopposite
effectat theselevels.
Eflects ol Lipoids on Viruses
The antagonisticeffectsof positiveand negativelipids were evident in
the sfudy of their action upon viruses.Generally,agentswith a positive
polar group appcaredto create favorableconditionsfor the development
of viruses,while those with a negativepolar group had an oppositeeffect.
This influence,which was first seenin phagesin vitro, becamestill more
evident in viral infections.
subcutaneousadministrationof positive lipids, such as sterols or insaponifiablefractionsof organs,inducedgreaterlocal receptivityto viruses.
In experimentswith smallpox virus in rabbits,for instance,virus inoculation of the skin induced an exaggeratedresponsein those areas where
positivelipids previouslyhad been injectedsubcutaneously
comparedwith
the responsein other previouslyuntreatedareas.In less sensitivespecies
such as mice and rats, positive lipid injectionsinduced abnormally high
local receptivity to virus inoculation. Intracerebral injection of sterols
followed by subcutaneousinoculationwith smallpox virus invariably produced nervous system localizationof the virus. Intraperitonealadministration of sterolsin very high dosesin mice prior to smallpox inoculation
produced a great degree of central nervous system localization. Intra-
L T P T D SA N D
LlPOlDs
/
155
cerebralvirus inoculation,after subcutaneous
administrationof high doses
of anti-fatty acids, brought death earlier in test animals than in controls
given intracerebralvirus alone.
A sniking opposite effect was noted for lipids with a negativepolar
character. In rabbits, subcutaneousinjection of a polyunsaturatedfatty
acid set up a local skin area refractory to smallpox virus inoculation,although inoculation was positive in other areas of the body. Death also
occurredlater,followingintracerebral
inoculationwith a neurotropicvirus
in test animals given subcutaneousor intraperitonealinjections of fatty
acids, than in controls.This partially protectiveeffect was oppositeto the
increased receptivity seen in animals injected subcutaneouslyor intraperitoneallywith insaponifiablefractionsand intracerebrallywith the same
virus, wheredeath appearedearlier than in controls.
The antagonisticeffectsof the two groups of tipids for viral infection
appearedinterestingfrom severalpoints of view. The cffects were local,
at the cellular level, where virusesthemselvesact. Subcutaneousinjection
of lipids inducedmanifestchangcsin responsetoward the virus in the skin
at the site of injection, and little or no changeat all elsewhere.we have
utilized this fact, as we will seebelow, to obtain information regardingthc
level at rvhich various agents act. A change induced in receptivity to
viruses,limited to the skin at the site of injection, woutclindicate activity
of the agentat the cellular level. Tests basedupon the skin responseto
smallpoxvirus infection have shown that, among the lipids with a negative
polar character,a maximum of influenceis exertedby the insaponifiable
fraction of organs of exodermicorigin from speciessensitiveto the virus.
The insaponifiablefractionsof rabbit skin and rabbit brain were the most
activeof the lipids tested.Among the fatty acids,the prevcntiveeffectwas
seen to increasewith the degreeof desaturation.It was almost entirely
absent in saturatedfatty acids, notably present in polyunsaturatedfatty
acids.
The increaseand decreasein receptivityof the skin to smallpox virus
following injection of lipids also furnishedinformation about the roles of
the polar and nonpolar parts of lipids in this specificactivity. An opposite
effect was seen between two groups of substanceshaving the same nonpolar group but differing in their polar groups.While the polyunsaturated
fatty acids of saffower oil, for instance,greatly reduced receptivity, the
same polyunsaturatedmembershaving alcoholsas polar groups increased
receptivity. The polar group--negative or positive-appears to be the
factor inducingthe oppositeeffect.
The role of the nonpolar group was studied by comparing saturated
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
f
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&
\
f
,
-{.'
\
Fto' 71. Lipids and microbes. Drawing of the progressivepassagetoward normal
bacilli of the tiny gram negative microbes obtained through the treatment of Bac.
Anthracis with fatty acids. The passagetakes place usually in successivesteps. Thc
gram positive formations appear first as fine granules;they later become clumps ancl
finally give the microbes their normal aspect.
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 ) .
the positive sterols.(Note JSl The sameinfluenceupon thc aging processes,
as manifestedin a prolongation of the lifc-span, was noted for polyunsaturated fatty acids with a long chain and evcn for some mcmbcrs of the
saturated seriesbut with a shorter chain.
Effects ot' Lipids on Complex Organisnu
Morphological changes-The same level separation was uscd in thc
study of the effectsof lipids on complcx organisms.Acting at chromosomal
levels, lipids led to the appearanceof monstrosities.various lipids, especially insaponi{iablefractions of organs, rvere injectcd into larvae of flies.
While an immediate change in the cells of the larvac could be traccd to
the subnuclear level as secn in chromosornes,monstrositiesrvere seen to
be induced in the resulting flies. A similar eJTectbccame evident when
lipids were injectedinto hens' eggsbeforeor during incubation.Especially
with cholesteml but also with insaponifiablefractions of organs, a high
proportion of chickens were hatched with spasticparaplegia.
The sarneproblem is being studied, in collaboration with P. Fluss, in
Drosophila melanogaster, grown for many generations in nedia to which
an entire scriesof difierent lipids from one or the othcr group are addcd.
This study is in progressand the resultswill be published later.
We have seen that the antagonisticeffects induccd by the two groups
of lipids could be related ultimately to oppositechangesin the fundamental
biological proces$ of aging. This appeared clear for lower morphologic*l
levcls of organization and was especially evident for cells. While anti-fatty
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
acids induce changes which can be regarded as corresponding to prolonged
youth, the polyunsaturatedfatty acids induce rapid aging with pyknosis and
karyorrhexis and death of the cells as old entities. This could be seen
clearlyin tumors,in which cellswith youthful characterlead to non-necrotic
tumoral ma.sses,
while cells that age rapidly produce necrosisin the tumors
followed by ulceration il the tumor is superficial.
The eftect of conjugated fatty acids was somewhatmore complex, indicating an abnormality in the induced processes.Their sdminisfixtion was
followed by the appe:uance of cytoplasmic and even nuclear vacuoles,
correspondingultimately to an anomaly of water metabolism.
The effect of lipids upon adipous cells appearedto be of special interest. The anti-fatty acids, especially the sterols, when injected subcutaneously in animals, induced a characteristicprocessin the adipous cells near
the injection site. These cells became very enlarged and higlrJy irregular,
with their content changed into an emulsion only slightly stained with
sudan. The fatty acids, on the contrary, imparted to adipous cells an abnormal resistanceto destruction. They remained persistently unchanged
even in the midst of very active processeswhich usually cause them to
disappear.Unchangedadipouscells were found encircled by the hvading
cancerousceus,deep within tumors in animalstreatedwith fatty acids.
On Pain-From the start of this research,the opposing effects of the
two groups of lipids upon pain has been most impressive. For the fatty
acids the degree of saturation is important. The saturated members of the
fatty acid seriesand even oleic acid are entirely without effect. Linoleic
and linolenicacidsshow a slight influence,while the polyunsaturatedmembers show a marked effect. Administration of highly unsaturatedfatty
acids and of acid lipidic fractions of certain organs,such as placenta,liver,
spleenor blood, uniformly decreasedpain of an acid pattern and increased
pain of an alkaline pattern. These oppositeeffectshave, from the beginning
of our study, contradictedthe idea that this influenceupon pain was the
result of the direct action of tlese agentsupon the nervous system.Furthermore, the opposite effects exerted upon the same pain by the other group
of lipids have confirmed the hypothesisthat the acrion takes place at the
level of the painful lesion, where the difterencesbetweentre two pains was
found to correspondto two oppositeacid-baseoffbalances.
In the study of the effectsof lipids on the pH of the secondday wound
crust,madein collaborationwith CarlosHuesca,we have demonstratedthat
lipids influence pain through changes induced on the acid-base balance
presentat the tissuelevel. The poeitivelipids constantlylowered this pH
while the negative lipids elevated it. (Note I Chapter V) Even more im-
L T P T D SA N D L T P O t D S /
151
portant than this temporary pH effect in establishingthe mechanism of
lipid action in pain was the change in the actual pattern of an existing
pain after administration of these agents. Polyunsaturatedfatty acids in
sufficient amount were found to convert an acid-pain pattern to an alkaline pattern, while sterols changed an alkaline pattern into an acid one.
We will return to this important fact later.
A pathogenic role for lipids becomesevident too, when pain can be
induced through the administration of lipids in previously painlesslesions.
Such lesions treated with large amounts of lipidic preparations became
painful. An alkaline pattern of pain was seen to appear after fatty acid
administration, while an acid pattern followed use of the insaponifiable
fraction. (Note 36)
At the tissuelevel,lipids also affectsuchacid-basesymptomsas vertigo,
itching, dyspnea,tremor, and even mental diseases.
In theseconditionsthe
same antagonismbetweenthe two groups of lipids-and the same opposite
effects upon the acid and the alkaline pattern---can be noted along with
the same possibility for changing the pattern to the opposite type if big
doses of lipids are administered.
wound Healing-The same manifest antagonism between the two
groups of lipids was also noted in their influence upon the evolution of
wounds. Changesin the sloughingand healing processwere followed by
measuring the size of wounds (Note 37) as well as by serial histological
examinations.The lipids with a negative polar group were seen to retard
the evolution of the processesby prolongrngthe first catabolic phase. positive lipids generally had an opposite effect. However, here too it could be
observedthat sterolshave relatively little effect on the healing of connective
tissue, but manifestlyfavor proliferation of the epithelia. This was especially evident in the changesin scar formation of the skin of treated animals. In rabbis treated with cholesterol,the epithelial scars were found to
have 8-10 layers insteadof the 2-3 characteristicfor the rest of the skin
and for the scars in control animals.
Regenerati6n-11 collaboration with E. F. Taskier we studied the
eftect of lipids upon the regenerationof liver in rats, after the resectionof
almost 3/q of. this organ. The rate of regenerationcould be estimated by
correlating it to the time of appearanceof fatty droplets filling up almost
all the cells. as a first step in the regenerativeprocess.In very young animals, this changein fatty liver cells was seento take place even within the
first 24 hours after resection.The change was progressivelydelayed as the
age of operated animals increased.In old animals the change in fatty liver
cells appearedonly after the fourth day.
162 /
REsEARcH rN pHyslopATHoLooy
The administration of lipids had a marked effect on appearance time
of fatty cells. Sterols induced precociousappearancein old animals. From
this point of view, sterol-treated animals appeared to react as young individuals, with fatty cells evident even on the secondday. The fatty acids
and acidJipidic fractions of organs showed an opposite effect, delaying the
time of app€arance of the fatty cells. Young animals treated with polyunsaturated or conjugated fatty acids showed no fatty droplets in the liver
cells for as long as three to four days. with higher doses of the same
agents,the fatty infiltration did not occur at all.
It is interesting to note a parallelism between fatty infiltration of liver
cells and the richness of adrenals in sudanophil subsrances.An almost
complete lipid depletion of the adrenals was seen after high doses of fatty
acids and coincided with a total lack of fatty cells in liver regeneration.
(Note 38)
organic Level-Effects of the two antagonistic groups of lipids at the
organic level have been studied in terms of manifestationsclearly associated with various organs. we will review these effects briefly here, with
more details to come when the therapeuticuso of lipoids is discussed.
lnlgslings-The influence of lipids upon intestinal function is marked
by the same antagonismbetween the two groups of agents.oral administration of large amountsof fatty acids,especiallyhigher unsaturatedsuch
as obtaincd from cod liver oil, was usually fotlowed by diarrhea. Diarrhea
also occurred after parenteral administration of these substancesin large
arnounts. It was interesting to note that parenteral administration of the
acid lipidic fraction of placenta, blood or even organs had a marked influence upon the colon and rectum in particular. High doses produced
tenesmuswith a mucous or even sanguinolentsecretion.This localization
of the effectsof the lipidic fraction appearedto be especiallyinteresting
from a therapeutic point of view, as will be seen later. The oral or parenteraladministrationof the oppositegroup of lipids, sterolsand insaponifiable fractions, has an opposite effect, a constipating one, which we will
discusslater together with its therapeuticaspects.
Kidney-The manifcst opposite effects exerted by the two groups of
antagonisticlipids upon diuresis raise the question of where these effects
take place. While a systemiceffect can be recognized,a more direct intervention upon the kidney also must be considered.The addition of the acid
lipidic fraction of organs,and especiallythose obtainedfrom pork kidney,
to the perfusion fluid in a dog kidney preparation produces a manifest
decreaseof excretedurine. The administrationof insaponifiablofraction
LtPrDs AND LTPOTDS /
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
corticoids,which made Seyleseparalethem accordingto their "phlogistic"
and "antiphlogistic" activity, would explain the two opposite manifestations inducing convulsionsand coma, produced in individuals previously
treated with the same anti-fatty acid agents.
On Cardiac Rhythm-The influence exerted by the two groups of lipoids upon the cardiac rhythm was studied under the same dualisl"icaspect.
The effects observed can easily be interpreted considering the role of the
differentiation of the cardiac cells for their part in the cardiac physiology.
The role of a cell in cardiac physiology is a direct function of its own
automatismwhich can ultimately be related to its degreeof differentiation.
The fact that the two groups of lipids acr antagonisticallyupon this cellular differentiation, the acid lipids exaggeratingit and the insaponifiable
fraction of sterols reducing it, has explained some of the effects induced
by these agents upon normal and abnormal cardiac rhythm. (Note 4l)
on oestral cycles-The action of the two groups of lipids at the organic level was also studied in the rat ovarian cycle. Daily, and even twice
a day, vaginal smears were made in animals treated with these agents.
when large amounts were administered,both groups suppressedthe cycle.
with smaller doses,only the lipids with positive polar groups, especiauy
sterols,produced this effect.
Systemic Level-Blood has appeared especially suitable for in vitro
and in vivo studiesof the effectsof the two groups of lipids at the systemic
level. The effects on different blood constituentswere analyzed and led to
very conclusive results. We will outline here the principal points of this
study.
Under the influence of anti-fatty acids, the erythrocytes become more
turgescent increasein volume, show a strong refringency of their crown
in dark field examination, and remain isolated from one another. The
sedimentationrate, if previously high, is reduced by treating blood in vitro
with insaponifiable fractions of organs. oxygen appears to be retained
longer in treated red cells than in controls.
The fatty acids have an opposite effect. Under their influerrce,the red
cells becomecrenelatedand develop a tendencyto form sludges.The sedimentation rate is increased.The color of the treated blood is dark and,
even after oxidation, rapidly darkens again. In vivo, lipids wittr a positive
charaoter induce leucocytosis,tlose with a negative character leucopenia.
This last effect is seen even in vitro. In Note 42, the influence exerted by
lipids upon the blood is presentedwith more details. (36)
On Temperature-The administrationof sufficient amoun{s of positive
lipids induces a frank elevation of temperature,while hypothermia follows
LrPrDs AND LtPorDs
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165
thc administrationof negativelipids. The relationship betweentemperature
and lipids, however, is not so simple since changesin external lemperature
influencethe balance betweenthe antagonisticlipids. For example, animals
kept in incubators at a temperature of 35oC show an increase in lipids
with a positive character.Animals kept in a cool place, such as a refrigerator, show an increasein lipids with a negativepolar group. The organisms
are able to combat the increaseof lipids with negativecharacter by means
of the normal defensemechanism,but are less capable of dealing with an
increaseof lipids of pcitive character.Therefore, while a high proportion
of animals kept in refrigerators adapt themselvesto the new conditions,
those in incubators die in a few days.
on systemic Patterns-The influence exerted by lipids upon various
other systemic manifestationswhich are reflected in abnormal patterns in
urine analyseshas been studied. In genora.l,the fatty acids induce patterns
corresponding to the offbalance of type D, while the sterols induce patterns
of the type A offbalance.Here again we must emphasizethat any lipid, if
administered in large quantity, influencesa.ll analytical values. A certain
specificity, however, is noted since, in relatively small doses,lipids induce
changesonly in certain values. Becauseof the inherent technical problems
conceming the patterns, only a few analysescould be followed accurately
over the period of time necessaryfor a clear recognition of changes in
small laboratory animals. It is for this reason that most of our studies in
this area were made on humans where pattern changes could be easily
identffied and followed over long periods.
It is to be emphasizedthat, under these conditions, the influenceof
lipids is exerted especially upon zrlreadyexisting abnormal patterns, increasingor decreasingtheir deviationsfrom the normal, or changingthe
patterns entirely. Abnormal patternswere induced through huge amounts
of lipids, which very seldomwere administeredto patients.Tnelp X shows
schematically the analytical changesinduced by the two groups of lipids
upon various urine and blood analyses,expressedas patterns corresponding to abnormalconditions,as well as upon the manifestations
presentat
other levels.
we will discuss these effects in more detail when describins the
pharmacodynamicpropertiesof lipids and lipoids.
Mechanismol the Lipidic Biological Activity
The analysisof the changesinduced by lipids has emphasizedcertain
characlerswhich appearof capital importancefor the understandingof the
biological interventionof thesesubstances.
In one kind of activity a lipid
rirt';:l;
l.';.rr
166
RESEARCH IN
PHYSIOPATHOLOGY
acts throughits Iipoidic properties.From the data concerningits distribution
in the organismit can be seenthat, due to its solubility characters,a lipid
introducedin an organismwill be selectivelyretainedby the existinglipidic
system.When such interventionthrough its lipoidic propertiestakes place,
the nonspecificcharacterof the activity of the lipid is prevalent.A second
kind of activity results from the bond realized through the charge ol the
polar groups.The positiveor negativecharacterof thesepolar groupsdeterTrsle X
LEVEL
EFFECTS OF STEROLS
EFFECTS OF F^TTY ACIDS
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
Urine Induceswater excretion
Inducessulfhydrylretention
Inducescalciumexcretion
lnduceschlorideexcretion
Inducessodiumexcretion
Inducesphosphateretention
Inducesretentionof surface
active substances
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
LIPIDS AND LIPOTDS /
167
minesthus the natureof this secondkind of activity.A third kind of activity
resultsfrom the chemical constitutionof the polar group, which will induce
selectivecombinationsand consequentlywill have a more specificinfluence.
A fourth group of changesare inducedby the activity which takes place at
the nonpolar group of the lipid and more specificallyat the energeticlormations presentin it. They will have a still higher characterof specificity.
with this systematizationof the activity of the lipids, a further systematic analysisof the influenceexerted by the lipids appears possible.
Through its selectivedistribution, the administrationof a substance
having lipoidic propertieswill influencethoseentitieswhich have lipids in
an active form in their constitution.The influence exerted will thus be
proPortional to the richnessof the entity in theseactive lipids. This fact
explains why the administrationof a lipid or lipoid affectsselectivelythe
abnormalentitiesrich in free lipids and to a much lesserdegree,the normal
ones. It is this selectivedistributionwhich will further limit the activity of
the lipoid to the lipidic systemand most manifestlyto the abnormalentities.
In the frame of this limitation, this activity resultsfrom the chargeof the
polar group. Similar effectsare thus obtained for all the different lipoids
which have the same electric positiveor negativecharacterof their polar
group. This explainswhy one can usedifferentagentsfrom the samegroup
and still obtain similar results.Agentschemicallyso differentas fatty acids,
mercaptans,persulfides,aldehydesor epichlorohydrine,have similar activity becausethey all have negativepolar groups.The characteristicof the
effectsresultingfrom the electricalcharacterof the polar groups,is that they
are common for the groupshavingthe samesign and diametricallyopposite
for the agentswith a positiveor a negativepolar group.
This effect was clearly seen in fatty acids in which the negativeciuboxylic polar group was changedinto the positive primary alcohol. The
biological effects of the new substancewere opposite.
It is in the third kind of activity that the chemicalnature of the lipoids
intervene.Certain effectsresultingfrom the bond of an amino polar group
will thus be differentfrom that of the alcohols,althoughboth act as positive
energeticcentersand as such have exertedother common effects.The same
is true for the carboxyland thiol groups.
Still more specificappear the effectsresultingfrom the interventionof
the energetic factors present in the nonpolar group, such as the double
bonds,and the energeticformationsthey realizesuch as conjugated,or two
double bonds separatedby a methyleniccarbon.
The various mechanismsinvolved have explainedfurther the different
kinds of biologicaleftectswhich result.The action of the lipid by meansof
168 /
nEsEARcH rN pHysropATHoLocy
the lipoidic effect will thus influencegeneral,nonspecificmanifestations,
suchas thoseconcerningthe permeabilityof membranes.only secondarily,
will thesechangesin membraneperrneabilityinfluencethe different metabolic processes
which the membranegoverns.
In the secondgroup of changes,relatedto the interventionof the polar
groups,the antagonisticeffectsinducedwere seento concern processesresulting from membranepermeability.It is only in a third change that a
more specificaction upon the differentmetabolicprocesseshas to be considered.These are concernedwith an interventionupon metabolitesor the
agentsgoverningthem. The characterof this last lipidic interventionis its
specificinfluenceexertedupon a definitemetabolicsystem.
we tried to interpretthe influenceexerredby a lipid or lipoid according
to the above systematization.
The recent developmentof the biochemical
methodsof investigationhas put into limelight many biochemicalprocesses
by consideringthem as isolatedmetabolicentities.Most of them were seen
to result from the interventionof enzymesupon more or less specificsubstrata. One of the principal objectivesof the actual pharmacodynamic
studiesis to correlateas directly as possible,biological eftectsof diftcrent
agentsto specific metabolic processes,most of them correspondingto a
change in an enzymatic process.This approach, while very interesting,
would not take into considerationthe important role played by the nonspecificactivity of lipids and lipoids. These nonspccificinfluencesthrough
changesin the lipidic systeminducedifferentchangesin different metabolic
processes.A nonspecificchangein membranepenneability will affect many
enzymaticprocesses.It explainsthe existenceof similar influencesexerted
upon theseprocessescommon to agentswhich have nothing more in common than their lipoidic propertiesand the presenceof a positiveor a negative characterof their polar group.It is this characterwhich binds an eftect
to the nonspecificintervention.This so-systematized
analysishas thus permitted to separatethe biologicalactivity of the lipids and lipoids, the more
specificfrom the lesserinfluences,and correlateeach one to a proper or
commoncharacterof the agent.This view has amply simplifiedthe study of
the pharmacodynamic
interventionof thesesubstances.
OTHER CONSTITUENTS
In addition to the chemical elements and lipids, other constituents have
been studiedfrom the dualisticpoint of view. Although the other constituents havc receivedless emphasis.interestinginformationhas been obtained.
I
..rr,.,,fra.
$tii.iti:t:r,:.;!$11;li:r1
LrPrDs AND LlPorDs
/
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*
I
!
170
/
xESEARcH rN pHysropATHoLocy
ency of thesesubstancesto act through metabolicchanges,than to a direct
influence upon the acid-basesystemicbalance. The first group acts as
heterotropicagentsand the secondas homotropic, as mentionedabove.
Abnormal Amino Acids
Our researchled us to severaltentativesto defineabnormal amino acids
and the proteins they form. one concernedtheir rotatory capacity. The
naturallyoccurringamino acidsare all levorotatory.However,the organism
constantlyhas enzymesableto attackdextrorotatoryamino acid membersas
if it would have to be prepared to encounter and destroy them. Such
dextrorotatorymemberscan be conceivedto appearon a statisticalbasisas
the result of the resonanceprocessseen to occur in all the synthesisin
nature. The interventionof specificenzymesagainstthem would have the
aim to control their existenceand especiallyto prevent their interventionin
further evolution. In a work hypothesisconcerningthe cancerousprocess,
we consideredthat their persistenceand especiallytheir participation in
forming hierarchic entities would correspondto the specific abnormality
characterizingthis condition.
In another work hypothesiswhich concernsalso cancerousprocesses
and which we will discusslater, abnorma.lproteinsare thought to appearas
a resultof the bond of a carbamicradical (295) to the amino acid group.
The resultingcyclic formation having the characteristicNCNC group in
it, would correspondto abnormal amino acids which would representthe
primary characteristicformation of the cancerouscondition. (Seechapter
II,Note I)
Carbohydrates-Glucose acts as an anti-fatty acid agent, possibly becauseof the glyceryl compoundsresultingfrom its metabolism.We have
studiedit in opposition to the respectiveacids-gluconic, glucuronic and
saccharic.Glucosehas an analgesiceffect,although limited, upon pain of
an alkaline pattern, and an oppositeeffect upon pain of an acid pattern.
The acid group has an oppositeeffect upon pain. This could be correlated with the changestoward acidosisseen in the local pH of the lesions.
A manifest changetoward acidosiswas seenunder the influence of glucose
in the second day wound crust pH. We have noted previously the role
playedby glucuronic acid as an agentwith anti-positive-lipidactivity. We
believethat it is largely through this mechanismthat it favorably influences
acid pain, having an indirect action similar to that of fatty acids.