Report of Nhi Program'activities Mweee 31mg - Forgotten Books

REPORT OF NHI PROGRAM'

ACTIVITIES

M WEE E 31mg

Medicine and experimental Therap eutics Branch

Section on Experimental Therap eutics

Section on Cardiody mmice

Branch 59 99

ea control Program 6 20

of Gardiove souler Physioloy 1 00

Cellular Physiology and Metabolism

of Chemisfiry of Natm l Products 1 58 w 1 73

of Chemical Phem colow 1 74. a . 225

of Clinical Biechemistxz? 226 a» 233

cry of Kidney and Electron-t 234, a»

cry of Technical Development

24 9

aces 2 a. 5Research Section

1 623 Center

art Institute

Sum m ary of Intram ural R esearch A ctivities

1 9 58

Laboratom of C ellular t siOIOQ and M etabolism Section on

C e llular Phxsioloa

T he followingm ajor research projects have been under inve sti

gation in the Section on C e llular Physiologr duringthe past year

T he correlation of structure with enzym atic function in

the ribonuc lease m olecule .

T he m echanism by which genetic inform ation is utiliz edin the biosynthesis of protein m olecules , with particular

em phasis on the biosynthesis of the proteins of bacteriophage .

Studies on the structure of fibrous proteins. includingm yosin.

fibrinogea and collage n.T he biom thesis of proteins in the herds oviduct.

O zadative phosphor-

fi ction and electron transport.

Structure and m etabolic role of lipOproteins .

1D The covalent structure of ribonuclease is now essentiallycom pletely known and it is possible to exam ine , in a rational way, the

re lationships between specific parts ofthe m olecular structure and the

enzym atic activity of this protein

. T hre e m ajor approaches to this

problem have been em ployed. in the first, a system atic study of the

reduction of disuli’ide bridges inribonuc lease has revealed that at least

one , and possibly as m any as three . of the four disulfi de bridge s in th e

m olecule m ay be c leaved by reductive cleavage to the SH form without

loss of all the enzym atic activity. O ne of these bridges. nam ely. that

connectinghelio cystinc til with helio cystine £

36 (countingfrom the N

terminal end of the m olecule) m ay be opened, and the SH groups so

form ed stabiliz ed by conversioa to their carboaym ethyl derivative s.

without loss of activity. it is certain that at least one m ore bridge m aybe opened with im punity and its identity is now under investigation by the"fingerprinting

"technique to be described below.

T he findingof m ajor simificance is that ribonuclease , in which

three of the four disuli’

ide bridges have been reduced (this derivative

havingabout 1 0- 1 5% the activity of the native enzym e). m ay be reoxidiz ed

with m olecular oxygen with the écom plete disappearance of SB groups . and

with the regeneration of 1 0070 of the original activity. P hysical studie s

show that this reform ed. native - like m olecule contains even m ore intra

m olecular coiling than the original substance , in spite of its norm al

activity . T he results suggest that the pre sence of the single rem aining

disuli‘

ide bridge is sufficient to direct the reform ation of a second critical

bridge , but that the third and fourth disulfide bond m ay reform in an ah

norm al way but without a de leterious effect on function. T he arrangem ent

of the disulfide bridges in the reoaidiz ed m olecule are now under investi

gation.

O i m ajor iniportance in these studies and in a num ber of the other

activities of the laboratory has been the developm ent of a siniple and re

producible technique for the rapids qualitative study of the general features

of structure in anunknown protein. Sam ples of protein are first dige sted

with proteolytic enzym es . T he resultingpeptide m ixture is then separated

into its com ponents by successive paper chrom atography and electrOphor

sale to yie ld a characteristic pattern of peptide spots . These m ay be

located by stainingwith ninhydrin and the peptides so located m ay be iso

lated and analysed for their am ino acid com position.

T his technique has been ot‘

considerable use in connection with

studies on ribonuclease discussed above . T hus , for exam ple . finger

prints have been prepared oi digests ofbovine , ovine and porcine ribo

nucleases of pancreas tissue and the resultingpeptides have been sepa=

rated, analysed and com pared with one another . C ertain regions of the

m olecule have been found to shout differences from species to species

and, in one instance , sheep ribonuclease , a lysine residue characteristic

of the beef ensyuie , has been replaced by glutam ic acid. T his m ajorspecies difference suggests , at once . that the particular area of th e

m olecule in which this variation occurs cannot be critically involved inthe bindingor hydrolysis oi ribonucleic acid. Sim ilar species studies

are now in progress on a ribonuclease from spinach leaves and it is

hoped to extend the work to ribonucleases from A g ergillus , E . coli,and Streptococcus .

T he location of the active center in ribonuclease is also beinginvestigated by the application of controlled proteolytic digestion. Som e

enzym es rem ove sm all portions of the peptide chain with inactivation

whereas others attach the chain Without inactivation . B oth type s of ev ents

yie ld valuable inform ation as to the location of the functional conste llationof am ino acids.

T he approach outlined above will, it is hoped, eventually giv e

sufficiently specific inform ation to enable us to begin a synthetic

approach to the active center of this enzym e .

2) R ecent advances in genetic techniques , particularly with

m icroorganism s and viruses , have m ade it clear that gene s consist of

a large num ber of loci, proban of an order of siz e corre spondingto

only very few nucleotides in the DN A . m o lecule . G eneticists hav e be en

able to construct m aps of the genetic m aterial of a num ber of organism s

in which a large num ber of such loci have been pinpointed W ithin a single

functional gene , presum ably controlling the biosynthe sis of a single protein.

One such study has involved the‘

l‘o even bacteriophage s and a

genetic m ap for the host range specificity function has been constructed

by Streisinger and his colleagues . T he protein in the coat of the virus

particle which is responsible for host range specificity has be en purified

in this laboratory to a very high degree and sam ples of this protein iso

lated from various m utants which have undergone m utations in different

positions alongthe correspondingregion of the genetic m aterial are now

being subjected to"fingerprinting. T his study is aim ed at determ ining

W hether or not there enists a direct correspondence betwe en changes in

the chem ical structure of the D N A"chrom osom e

"of the phage and the

am ino acid sequence of the protein whose synthesis is under the control

of this region. A nother protein in the coat of the virus particle has todo with the penetration of the viral DN A into the host bacte rial c ell.T his protein has been identified as a lyso zym e and has been shown to

be sim ilar in m any of its properties to the corresponding enzym e in eggwhite and in various other tissue s . Lysosym e has been isolated inreasonable quantities from bacteriophage T 2 and from lysate s of E . coli

produced by phage infection. A search is now under way for a technique

by which the genetic loci in bacteriophage T 2 m ay be m apped so that the

study of correspondence between genetic inform ation and lyso zym e

structure can be undertaken. in paralle l with the se studies , a system atic

investigation of the structure of eggW hite lyso zym e is beingpursued in

order to obtain a baseline for com parison of the enzym e from the bacterio

phage . T hese studies also involve the fingerprintingtechnique describedabove .

(3? A s a consequence of studies on the three- dim ensional strum

ture of polyproline and of promos-glycine copolym ers , inform ation was

gained which has contributed considerably to the understanding of the

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4 ) Studie s on the m echanism of protein biosymheeie in the

hem e oviduct are being continued. Dr . H andler hae m ade considerable

progress in the isolation and characteriz ation of the intereettng lipid»

soluble substances which appear to be good candidates: as toterm ediete e

in protein bioeyntheete in this tissue . F ive distinct fractiono can be

separated by the use of alum inum silicate comm a and countercurrent

dietetbutioo mad the fractions contain very large am ounts of radioactivitywhen isolated teem tissue which has been incubated with radioactiv e

em mo e otde . T heee etodiee ere of greet tetereet in View of the lipidnature of the eodopteem tc reticulum which ie 8 0 like ly to be involved inthe e eeem bly ofprotein m olecules . to the course of the se studies D r .

H andle? hoe e leo retoveettgeted the technical problem s involved in thecounting ot

‘

C M and M 8 developed mtereettogm odifications of the theortee

originally proposed by W , Libby and othere for the oelfa eboorptton phenom

w on,

59 Dre , titelley and B rook heve m otioned their etudiee on the

nature of oxidative phoephoryletton alongthe lim e of le ot yeerfle won't-l o

T hyromtn and thyrow analogues havinghorm one activity have been

ehown to concentrate oetdetioe Without alteringphoephoryletiog efficiency .

A t higher comeeoteettone mad followingpeetoetabetton, anem ones; W e e ob »

serv ed. T he obeew ettone ho ve eoggeeted that the uncoupling effect to e.

eecondery one but it is still onem m Whether the acce leration of oxidetloo can be regarded ee phyetologteeuy otwtt

’

tceet. A num ber of V itam in

K and V item tn E m elogeee have eleo been exem tned in the m bm itochondrtet

eyetem . Som e ot‘

these ceased m arked acceleration of phoephorylettoe and

are beingfurther mveettgeted,

6) De . R odbell he s exemto ed the alpha. and bets», ltpOproteim m ole ea

males to peeelletwith the proteins W atch m e preeent m ohylom toroe e .

One of the m ajor com poneote to chylom icroo e he. been shown, by the

fingerprintingm ethod, to be tdemttcelwith the Mgho deoeity lipOprotem

of pleem e . Stodiee with C m ~lebelled am ino e etde have todieeted that the

alpine lipoprotein of pleem e is not to equilibrium with the correepondtngchylom tcroo protein oed that the ohylom teeon em ttaeeie m ust involv e a

eepe rete pool ofm m particular protein eubetemce .

T he highc demetty lipopeoteme m ey be m ade soluble in aqueous

system s by rem oval of the liotd com ponents , fitmdiee are under way to

investigate the"re a eymttneete

"of alpine lipoeroteine from its com ponent

paste l D r. Rodbell, together with D r , F redmickeoo ot‘

the Section on

Metabolism . is also active ly investigatingthe m etabolic origin of the

chylom icron proteins and high- density liponroteins using radioactiv e

techniques .

Duringthe past year the Section on Enzym es has been concerned

with studies on the enzym atic m e chanism s of the followingbiochem ical

processes: 1 ) the m etabolism of betesocyclic com pounds; 2) the m etano

lism of three carbon com pounds; 3) anaerobic oxidative phosphorylation

and e lectron transport; 4) cellular differentiation and protein synthesis;

5} the m etabolism of isoprenoid derivatives; 6) bom oserine m etabolism .

It W i s «W z R iboflavindegradation E , R . Stadtm an, P . Sm yrniotis , T . Miles) . Several into ).M

m ediate s in the oxidation of riboflavin to carbon dioxide and am m onia

have been isolated from the culture m edium of an aerobic bacterium

grown on riboflavin as its m ajor source of carbon, nitrogen and energy.

Detailed chem ical and enzym atic w aigete oi tbese com pounds revealsthe existence ot

’

at least two m ajor pathways of riboflavin dissim ilotlon.

One m echanism involves an initial ozddative c leavage of the pyrim idine

ring (ringC ) with tbc form ation of stoicbiom etric am ounts of lo ribitylo

2, 3- dilteto a-l, 2, 3, d tetrabydsoo o,

°7 -dim etbq ninonaline (com pound

urea, and a one carbon derivative (probably T his is a cat ions

transform ation since all of the m ost reasonable postulated interm ediates

have been synthesised tested and found not to be m etaboliz ed. C om pound

l is subsequently converted to a second closely related com pound (com

pound m by a synthetic r eaction which involves rem oval of two oxygen

atom s from the ribityl side chain and the addition of two cannon atom s

and one nitrogen atom to the side chain. T he exact structure of com pound

H is yet ondetem ined. C om pound H is subsequently conv erted by a series

of unknown reactions to 3, 4 dim ethyl- ai-pyrone o S- carbosyllc acid which ,

in turn, is ofi dised to 0 0 2. A second pathway of riboflavin breakdownis indicated by the isolation at other m etabolites which are oxidiz ed to

0 0 2 and M 3 witbont the interm ediary form ation of any of the above

m entioned substances. P reliM nary evidence indicates that this pathwayinvolves an initial attack on ringA of the riboflavin m olecule with theform ation of a pteridine derivative . One snob m etabolite has been foundto replace riboflavin forgrowth of a riboflavin requiring strain of lacto »

bacillus » suggestingthat it m ay be an interm ediate in riboflavin biosyn~

thesis .

29 T he m etabolism of three carbon comm as .

P rogionic acid oxidat'

fl tnr . V ageios) . A new pathway for

the biological oxidation of propionic has been elucidated in studies with

cell-free enzym e preparations ofthe anaerobic bacterium C . klufl erio

T his m echanism involvee the interm ediary form ation of propionyl C os. »

acrylyl C oA , fl -hydroayo

propienyl C O A . m alonyl sem ielciehyde 0 0 A ,

and m aionyl C oA . T he fate oi m aioayl C oA is still uncertain but som e

re sults indicate that it m ay he deearboxyiated to acetyl C oA and 0 0 3,

T hese results supplem ent those pz'

evious findings (reported last year)of lactoyl C os. and F - a1awi Goa e s activated interm ediaries in the

m etabolism of propiohic acid. P roof for the biological form ation of

these activated three e eeeee com pounds directs attention to their poesibie

role in the eaergy m etabolism of diverse bioeyathetic processe s .

iht and aiaoie e me s . H . Go ldfineend E . R 0 atalyse

the cem rer and

propieaic acid. Deteiied studies ofthis ferm eeztation have revealed thesurprisingfact that pm vate

'

is act an interm ediary is the oxidation of

alanine , Duringthe coarse oi theee stadiee it was discovered that with

c ell-free extracts pyruvate is utiliz ed for the eyethesis of a m ajor m etah v

e lite which has been ieoieted amid tentatively characterised as a hydroxya

dicerhoayam iao acid tpeohehiy y- am leo »

7a m ethyi

n

glutam ic acidic

Studies are in progress cm the m echaeiem oi this reaction and

the role of the new am ino acid m etabo lite in. the m etabolism of this

orgeaiem o

33 fe m W .

“ o

dynam ic considerations. it has been pestuiated that the energy for grewth

of the anaerobic bacterium C . him eri is derived from the reduction of

crotohyZio C oA te hatysyl O oh by reduced diphosphopyridine nucleotide .

T his reactioa is aseocieted with a standard free emerw change of about

0 1 4 0 000 calories A recent report hes claim ed that in soluble c ell free

preparations of C . hisfl ee phoephoryiation is coupled with the above

oxide- redaction eroceae o E xtendingw e own etaciiee in?this biochem icalreaction, the occurrence of phoephorylatioh in the anaerobic m etaboliem

oi cretoayi C oA and re lated thioieeter derivetives has been confirm ed;

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c lostridium capable of utilizingthis substance as its sole energ

and carbon source . C ell- tree extracts of this organism have be en

shown to catalyz e the conversion of choline to one m ole of trim ethyl

am ine and one u half m ole each of acetate and ethanol. Extracts of

this organism were found to contain a cytochrom e pigm ent which is

spectrally sim ilar to anim al cyto c hrom e C o T his discovery is the

first exception to the previons generalisation that c lostridia do notcontain cytochrom e pigm ents . A ltnongnthe exact role of this pigm ent

in the m etabolism ot‘

choline is not yet known, it is of special interestthat in order to serve as an electron carrier under the anaerobic condi

tions of m etabolism , it m ust function at a redos potential W ell belowthat ot

‘

the anim al cytochrom e system . The biochem istry of this pigm ent and the possibility that anaerobic dissim nlation of choline iscoup led with phosphorylation are im m ediate topics of further study ,

4 ) H om oserine m etabolism tDr . F lorin) . O -phospho

-onom o »

in the isom erisstion of hom osertne to threonine in certain m icroorganism s

F rom the m ecnm istic point of V iew the role ot s. phosphate ester in such

an isom erisation is of special interest since there is no established

precedent for such a reaction in m etabolism . it is of furthe r interest

because it m ay be analogous to , and serve as a useful m ode l for , an

early step in the biosynthesis of steroids; Win. in the conversion of

m ovalonic acid pyropnospnate to A 3~iscnentenol pyrophospnatelo

T he enzym e catalyz ingthe conversion ofpho sphonom oserine

to tnreonine and orthwnospnete has been extensively purified fromextracts of N eurosgors and was shown to reqe lre pyridoxal phosphateas a coensyrne . Stndtes ofthis reaction in tritium - labe led teeter reveal

that no significant incorporation of tritium into tnreonine occur s . T his

e lim inates from further consideration one postulated m echanism invo lvingthe interm ediary form ation of an olefinic derivative such as vinylglycine o

tdt T he re c'd

. sf M e ow»? DP ).

m olecular hgdrogen (Bre e S, Kinsley and E . R , Stadtm anl, Two heat

stable coensym es are involved in the reduction of DP N by m olecularhydrogen as catalysed by soluble extracts of C . sing er

-i. One of these

coenzym e s was identified as flarine adenine dinncleotide (F A D ) . P uri

fication of the second coensym e"

is still in progress . A s judged by behavior on ion exchange resins, electropnoretic m obility and sdsorbsbility

on charcoal, etc . it concern to be a strongly anionic substance probablyofnucleotide nature . The discovery that F A D is an obligatory coenrym e

in the reduction of D P N by hydrogen is of spe cial intere st since thestandard redox-potential of free F A D is well above that of eitherhydrogen or D P N . T herefore , from therm odynam ic considerations

it seem s probable that in the hydrogenase system the potential of the

F A D is m arkedly reduced perhaps by com bination with the ape-enzym e .

T he probable existence of such low-potential flavin system s require s

a t e - evaluation of the concept that fi avin enzym es , because of their

high potential, are involved only as electron carriers in term inal

re spiration.

5D T he oi ellular dif eren ttia ion and pe tr ein

sfl thesis (Dr. B . Wriwtb. The dramatic changes in the m orpholowand m etabolism of the slim e m old duringthe process of ce llular ditierentiation are associated with m arked changes in enzym e com position

and in concentration of the free am ino acid pool. A kinetic analysis

shows that as differentiation proceeds , first, the siz e of the free am ino

acid pool decreases to one o third its initial vsalute , then the alcohol-assh ole

proteins decrease and iinslly between the singand fruit stage s , the re »

m ainingprotein fraction decreases . The greatest proteolytic activityoccurs after the free am ino acid pool reaches a ion level, at which tim e

m any enzym e activities are at their highe st. T he results are consistent

with the workinghyp othesis that a decrease in m ine acid pool, broughtabout by starvation, m ay initiate endogenous protein breakdown.

Evidence has been obtained showingthat acrs sin, the chem o

tactic steroid- like horm one which is needed to initiate aggregation of

am oehae duringdifferentiation is not involved as a coenzym e in the

D P NH o T PN H transhydrogenase system nor does it have any direct

effect on term inal respiration.

6) T he m tabolism of iso -

_

eri ativee . C itronellol is a

low m olecularWeight compound (2isoprene omits and it serves as an

ideal m odel com pound to study isoprenoid degradation T herefore in

order to facilitate studies on the biochem istry of polyisoprene m etabo o

liens, an aerobic bacterium o ne isolated from soil enrichm ent that can

utiliz e citronellol as its sole carbon and energy source . Studies with

cell-free extracts of this bacterium were found to contain all of the en

zym es previously established in the biosynthesis of acetoecetate , ,8

hydrosybntyrate , and fl o hydroayo fl - m ethylglntaryl C oA , and in term inal

respiration by the T C A cycle . H owever, dosingthe oxidation of acetate

by cell suspensions onto‘

zii‘i'

i of the acetate carbon is utiliz ed for the biosynthesis of a relics? substance which has been isolated and tentatively

identified as a terpene . It is further dem onstrated that duringtheoxidation of citronellol or isovaleric acid. extensiv e incorporation

of C1 40 2 into acetate and fi -hydroxym ethylglutaric acid occurs .

T he detailed m echanism s of citronellol cateholism and the biosyn

thesis of the terpene- like com pound form ed from acetate is being

pursued further.

- Section on Metabolism

T he research activities of the Section on Metabolism for the

year 1 958 are best described under several categories

studies on the basic physiology of fat absorption and transport;

studies oi”

the iactors controllinglipoprotein levels. includingin vitro studies of lipoprotein hiosynthesis and the effects of

various agents each as dietary fats;

studies on the m echanism s of protein biosynthesis;

studies of clinical and experim ental nephrosis. includingim m nnochem ical studies;

basic stadies on protein structure;

studies on plasm a protein degradation and excretion.

ll M . -

th-bem 9 1 a . of ire-me e ts

P revious studiesin this laboratory have estahlishedthe importance of

unesterit’

ied fatty acids (UF A Din the transport of fat from tissue depots

to sites of omdation. it has now been shown in m that there is verylittle difference in the rates at which different fatty acids (palm itic .

oleic and linoleic acids) are converted to carbon dioxide . E xercise

m arkedly increases the rate oi conversion oi circulatingUF A to 0 0 3

(6- 1 0 feldpo

T he previously reported action of epinephrine in elevating serum

UF A concentrations has been further stadied. T he elevation of serum

UF A followingepinephrine is not m aintained even when large am ounts

of epinephrine are adm inistered intravenously. The return ofUF Alevels toward norm al occurs at shoot the sam e tim e the blood glucose

levels rise . in separate studies it was shown that adm inistration of

large am ounts at glncose conld prevent the epinephrine- inducedUF A

response . Studies on adipose tissne in vitro shots that this is a direct

and phospholipids (about 50 per cent above control values) . T he

possible im portance of this observation in relation to stress - induced

hypercholesterolem ia is beinggiven serious consideration .

T he use of inhibitors of chole sterol synthesis to effect a lower

ing of serum cholesterol levels has been further explored . Several

com pounds structurally related to A4

« cholestenone have proved in

effective as inhibitors tz-alpha-m ethyl- cholestenone . S- alpha

- m ethyl

cholestenone . sitostenone and 3-m ethoay- cholesterol) . It has been

previously postulated that the effectiveness of A 4 - cholestenone m aydepend upon its conversion to dihydrocholesterol. T he ineffectiveness

of the substituted cholestenone derivatives m ay possibly be attributable

to the inability of the organism to reduce them . B enzm alacene (bl- C lm ethyl 2, 3

- dic p- chlorOphenylpropylm aleem ic acid) has been shown to

be effective in loweringcholesterol levels in dogs up to 50 per cent,

confirm ing studies by M ercia. Sharp and Dohm e . C linical studies on a

sm allnum ber of patients indicate the drugto be effective but gastric

irritation and elevation of E SP retention appear to be problem s . ME R

29 l-tp- tfl- diethylam inoethoxyl~phenylle lo tp

e tolyll- 2- tp- chlor0phenyl)

ethanol) profoundly influences chole erolm etabolism in the rat, causingthe serum cholesterol levelto dropla% low as 1 5 m g%. and decreasingthe cholesterol content of the liver by 50th. A none ssponifiable lipid

distinct from cholesterol accum ulates in the liver of treated anim als

and the nature of this is under study.

A new m ethod for incorporatinglipids into lipoproteins in vitro

has been developed. T his sim ple proc edure perm its the incorporation

of labeled com pounds into lipoproteins at high specific radioactivity and

rushes it possible to carry out tracer studies with a better approach to

the truly physiological state . in addition it prom ises to be of value in

the study of lipoprotein transport of carcinogens and other trace com

ponents .

3» sm ile Studies

on the incorporation of am ino acid analogues into crystalline proteins

have been com pleted. The conclusive dem onstration of such incorpora

tion has im portant im plications with respect to the specificity of protein

biosynthesis . Studies of low m olecular sleight peptide derivatives

occurringin rat and lipid liv er are continuing. a large fam ily of low

m olecular weight peptides associated with an acidic com ponent. probab lynucleotide in nature . has been dem onstrated by m eans of chrom atography

on cellulose colum ns . C onclusive proof that the se are involv ed in

protein biosynthesis has not yet been obtained.

in the course of these studies a m ethod for the radioassay of

tritium labeled proteins in the liquid scintillation spectrom eter was

developed. A m ore general m ethod for countingaqueous solutions in

the liquid scintillation spectrom eter has been found and is beingfurther

explored to m ake it routine .

to the general clinical im pression. there are a significant num ber of

nephrotic patients in Whom the elevation of serum lipids is prim arilyin the 13 l

o lipclprotein fraction rather than in the very low density lipo

protein fractions . T here appears to be a spectrum of lipoprotein

patterns and duringtherapy with either steroids or album in the lipopro

tein pattern is observed to change from one typ e to another . The se

findings cast doubt on the hypothesis that the nephrotic suffers prim ar ilyfrom an inability to convert v ery low density lipoproteins to higher densitylipoproteins . While the effectiveness of intravenous serum album in in

loweringthe lipoprotein levels in nephrotic patients has im plicated al

bum in specifically. eaperim ental studies in the rat show that dextran

is to som e extent able to duplicate these effects of album in and this

raises question as to the specificity of the album in m olecule in this

re spect. A ttem pts to dem onstrate antibodies to hum an kidney in the

serum of patients with renal disease have been negative . On the other

hand. usingthe saline tannic acid hem apglutination test, it has been

possible to confirm the presence in the sera of patients with thyroiditis

of antibodies against hum an thyroglobulin. Rabbits im m uniz ed with

rabbit nucleoprotein extracts develop antibodies both against these

hom ologous nucleoproteins and '

against hum an nucleoproteins .

Forty- five patients with the nephrotic syndrom e have now been

treated with adrenal steroids and partial or cataplete rem issions have

been obtained in alm ost tnoo thirds of these cases .

52) W hidii e on reacte _

st cture . Kinetic studies of the

digestion of m yosin by proteolytic enzym es reveals two distinc t rates .

Sim ultaneous study of the optical properties of the m olecule suggest

that the portions of the m olecule not involved in hells form ation are

the parts rapidly digested. P relim inary studies suggest a sim ilar

phenom enon in the case of collagen.

Laboratory of C hem istg of N atural P roducts

T he following sections sum m ariz e work in this laboratory.

A . Isolation Studies . T he principal isolation study in progress

duringthe year was concerned with the hum an m acrom olecular vaso

dilator "kallikrein. It is currently believed that kallikrein is a pan

creatic product. present in plasm a and in urine . that acts on a norm al

protein substrate thallidinogen, a com ponent of the eta-globulin fraction)

to yield a dialyzable polypeptide (kallidin) with profound vasodilator

properties . T he kallikrein is held in com bination in the circulatorysystem by a protein inactivator and its vasodilator properties are

exerted only when this com plex dissociates to give free kallikrein;when this occurs the action is m ediated through the polypeptide kallidin.

T he physiologic significance of these relationships is not known. but it

is now clear that there are perhaps several kallihrein- like substance s

present in the hum an.

T his problem is being studied tDr. P ierce ) in collaboration withD r . S .JSarnoff and his colleagues . T he chem ical work is concerned

with the problem of isolatinghallihrein and its substrate kallidinogen.

and ultim ately kallidin, so that they m ay be defined as chem ical individo

uals . A particular need is to isolate kallidin for study and to determ ine

whether the several com pounds ofthis type described in the literatureare identical. T he technical problem s are quite difficult; the isolationof protein or polypeptide m aterial in a high state ofpurity usually re

quires extensive fractionation, and in this case relative ly sm all am ounts

of m aterial are available . Several procedures have been studied for

kalliltrein or hallidtnogen. T he heat results were obtained with a com a

hinatioa of FEE - 64 resin and DEA E ce llulose . H ogpancreatic kallikrein

(P adutin) was also studied, but the m ajor part of the work has been withhum an m aterials . R elatively good fractions have been prepared. butthese are not yet pure enough for characterisation by chem ical m eans ,

and the present work is concerned with getting still purer sam ple s of

hallihrein arid hallidinogen.

T he physioloc eaperim eots and s ortswith hypotensive patients

were carried out in D r . Sarnoit“s laboratory. A urine assay procedure

was worked out and has been puhliehed.

The urine base project (Dr. F ish) was term inated in July. Seven

bases ofunknown structure rem aia from this trot-h i

T he alkaloids of H im antandra bel%raveana were isolated in an

Itattem pt to throw som e light on the Kuru disease of. N ew Guinea.

was found (Miss Z altzm anlthat himbacine was a physiologically active

agent. producingconvulsions endingin death. and that this action dupli~

cated that observed for the crude plant extract. which was reportedlytaken orally in N ew Guinea. T here is an A ustralian chem ical groupworkingon this problem at the pre sent tim e . and it is unlikely that astructural study of the alkaloid will be pursued in view of the A ustralianwork .

Several hum an m etabolites of unknown structure . pre sent in

instances of anem ia and m alabsorption. were studied in conjunction

with D r . C . E . D alglie sh of the P ostgraduate Medical School. London.

T he isolation wort; was carried only far enough to fix the identity of the

substances as skatole m etabolites. and further work was directed to

exam iningthe biological and chem ical reactions invo lved in the form a

tion oi‘

these m aterials .

A problem directed to the isolation of the glycolipids of hum an

red cells was started recently (Dr . Sweeley. D r . Moscate lli) . T he se

com pounds are rather com plex substances m ade up of sphingosine (or

one of its relatives) and a long- chain fatty acid which often carries an

af-hydroxy group . glucose or galactose . and usually neuram inic acid.

A distinguishingfeature of these com pounds lies in the fatty acid part;

acids such as cerebronic. ne t-“sonic and lignoceric are reported to be

present, and it is be lieved that different tissues contain different acids .

One of the problem s lies in findinga way to deal with cfi-hydroxy acids .

so that they m ay be studied by gas chrom atographic techniques; another

lies in deve lopingm ore effective ilolation m ethods for glycolipids thanhave been used previously. T he present work is on isolation m ethods .

B . c R eaction . Studies in this area

oth chem ical and biological work dealingwith biological transform ations. with the aim of establishingthe basic chem istry involved ineach instance . W e erode studies were continued. Evidence was found

pointingto the existence of a cellular am ine oxide rearrangem ent reac

tion (Dr . F ish. D r. Sweeley) . The stereochem ical aspects were studied

through the preparation of both diastereoisom ers of nicotine oufde (Dr .

Su eeleyi; one underwent dem ethylation readily. the other did not.

Microbial dem ethylation atomes did not lead to a definitive answer on

the ability of the omda to replace the am ine in a biological dem ethyla

tion reaction; this m ay be due to the inability of the oxide to enter

particulate structures (the two previously observed biological dem ethyl

ation reactions for the oxide were found to occur in the so luble portionof a ce llular fraction) .

Studies on the catalyst requirem ents and on the basic chem ical

m echanism of the reaction have been started in A ustralia by Dr . Cym er

m an C raig. and these will be continued in B ethe sda. T he effectivene ss

of the catalytic effect depends on the nature of the iron- coordination

com plex.

Studies on the m etabo lism oi”

tryptam ine (D r . F ish) have been

term inated. A new product found and characteriz ed by chrom atographic

and electrophoretic m ethods was quite sim ilar to N -form yltryptam ine

in its properties. but a definitive structural assignm ent was not m ade .

T he problem is currently under study by D r . Jepson in D r . Udenfriend°slaboratory.

T he oxidation (hydronylationlof tryptophan derivatives was

studied to gain additional chem ical knowledge of the reactions of these

com pounds . it was found that the m ajor product of hydroxylation (ironascorbic acid system ) was not the d- com pound. but an isom er. This

reaction is under detailed study for shatole . Oxidation at the 2 and 3°

positions also occurs . The bio logical products are under study and

com parison with synthetic products. with the aim of determ iningthe

nature of the m ajor biological hydroaylation route . it is not the 5

pathway. hydronylation followed by conjugation. and ring- opening

followed by hydroaylation and conjugation are the two chief reactions

in the hum an and in the rat. o -A m inoacetophenone . a lipid- solublearom atic amine . is one oi the biological interm ediates .

C . Structural Studies . The m ajor work in this area has been

concerned with the A m aryllis alkaloids tDr . Wildm an. D r . F ales , Dr .

Uyeo. D r . lnubushil. T he worb of this group has established m ost of

the current chem ical istructurallknowledge of these com pounds . T he

isolation studies have been relatively straightforward and m ost of the

work has been devoted to establishing individual structures and struc o

tural relationships . A num ber of new structures were determ ined

duringthe year. and an observation of considerable chem ical interestwas m ade in regard to cyclic strained lactam s . T heoretical predictionshad been m ade about the properties of certain specific lactam structures.

but since none oi these had ever been observed. the problem was one of

discussion rather than experim ent. A situation of this kind was found

- 21

Laboratogz of C ardiovascular t siolofl

A n attem pt was m ade to exam ine in broader perspective the

im portance of the relationship generally known as Starlingfls Law of

the H eart for the regulation of the intact circulation. T he ventricular

function curve concept was em ployed. More specifically, canine ex.

perim ents were designed so as to ascertain whether nerve pathways

could shift the baseline of the relationship between fillingpressure and

external ventricular stroke work . T o do this . m easurem ents were m ade

of atrial., arterial and ventricular pressures while m etering cardiac out»

put. P ressure and flow were also m easured in the independently perfused

carotid arteries with exclusion of other arterial supply to the head. H eart

rate was held constant in all experim ents by atrial stim ulation. (meet s-

a

tions were m ade which dem onstrated that stim ulation of the isolated left

stellate ganglion shifted the ventricular function curve to the left and that

the extent of the curve shift is a function of the frequency and/or voltage

of the applied stim ulus. Stim ulation of the distal cut end of either vague

produced the opposite effect. that is. a curve M t to the right. T he

order of m agnitude of the changes observed was large . T he se superi

m ents m ake it po ssible to put Starling0s Law of the H eart in clearer

perspective . T hat is. the centralnervous system has available to it

efferent pathways by m eans ofwhich it can system atically m anipulate

the relationship between fillingpressure and ventricular stroke work .

Further experim ents were then done to ascertain Whether the se

efferent pathways are enercised by changes in carotid sinus pressure .

Stim ulation of the carotid sinus nerves produced hem odynam ic response s

identicalwith those deserved when withdrawing sym pathetic stim ulation

or initiatingregal stim ulation. that is , a shift of the ventricular function

curve to the right. E levation of the carotid sinus pressure had the sam e

effect. Loweringthe carotid pressure shifted the ventricular function

curve to the left. The observed changes were m arked. In sum m arythese data dem onstrated that the organism has available to it pathways

by m eans ofwhich the heart is caused to contract m ore forcefully at

any given fillingpressure when arterial pressure is low and. conversely,

is caused to contract less forcefully at any given fillingpressure when

arterial pressure is high .

It was further observed that. with large changes in carotid

pressure . totalperipheral resistance changed one to two fold while

changes in ventricular externals ort: increased m ore than ten fold at

the sam e or even lower fillingpressures . T hese data suggest that

the dom inant consequence ofbaroreceptor stim ulation for circulatoryregulation is the reflex effect on m yocardial contractility rather than

on peripheralvascular tone .

investigations have been initiated to determ ine whether thesenervous pathways are im portant in circulatory regulation in exercise .

While , as described shove . attem pts were m ade to relate the

intim ate perform ance characteristics of the heart to the overallbloodflow requirem ents ofthe organism , experim ents were continued of the

type aim ed at elicitinga m ore precise appreciation of cardiac energetics

p31; 5 3 . G enerally speakingthese 1were of three types .

The first utiliz es the m oving of a lever by the m yocardium in

relation to a second fixed point which does not m ove . T he change in

distance between these taro points is siwaled to a recorder through a

low-inertia. m icrotorqae potentiom eter . T he recorded changes in"fiber

length , when exam ined in relation to the pressure events of the cardiac

cycle , by and large appear appropriate , e .g. fit our preconceived notions

ofwhat is occnrring. The principal interest in this m easurem ent is to

be able to follow changes in diastolic fiber length. systolic fiber length ,

and rates oi’ change in contraction and relaxation. With the other sim ulc

taneonsly recorded param eters it appears that we are able to exam ine

for changes in end -diastolic distensihility under the influence oi’ both

cardiac sym pathetic and nasal stim nlation to ascertain Whether any such

chm gee are doe either to the specific inflnence of the nerve stim ulation

or changes in the duration of diastole or hath . E ventually it is hoped to

exam ine m ore in detail the supposed relationship between end-diastolic

fiber length and m yocardial 0 3 cercisnM ptiono

T he second type of experim ent in this group resulted from an

incidental observation that fillingpressure rose and ventricular work.

fellwhen the site oi electrical encitation eras ch i-aptly changed from

atrium to ventricle . A system atic stndy was initiated to study this

phenom enon. This consisted of the m easurem ent of pressures in left

and right atria, pnlm enary artery and sorts and, in som e experim ents .

also in the left ventricle . T otal aortic flow «C . 0 . m inus coronary flow)was also recorded and data obtained for the construction of ventricular

function carves . it h as observed that the change from an atrial to e.

ventricnlar site at excitation pit

odnced a prononnced shift of the ventricu

lar function curve to the right, that is . less eaternalwork at any given

fillingpressure . In m any instances. this effect was such that therewas m ore than 50% decreased in externalwork at the sam e filling

pressure . It was also observed that m yocardial oxygen consum ption

rose when goingfrom atrial to ventricular stim ulation which. since

externalwork fell. resulted in a decrease in m yocardial efficiency .

One possible explanation for the observed phenom ena being enter

tained is that the contraction of ventricular fibers is less synchronous

when one excites what is essentially an ectopic focus than when the im

pulse is propagated norm ally. in support of this view are the observa

tions by other. that the electrical propagation of an innpulse originatingin the ventricle is slower than an im pulse of atrial origin. We had a

further strengtheningof this interpretation from our observations that

the upslope of ventricular pressure is less steep as is the curv e showingthe rate of fiber shorteningas m easured by the newly developed length

lever. T hat is. if the im pulse is less W ell coordim ted’

and propagated

less rapidly, the initial and last fibers to contract are doing so with

the red er of the ventricle actingas a flaccid aneurysm m ore than

would be the case when the contraction is m ore synchronous .

A n additionalm echanism which appears to be involved in theobserved results is the asynchronicity (relative to norm al) of the se

quenes of atrial and ventricular contraction such that the ventricle is

less abundantly tilled when the atrium contracts against a closed m itral

valve .

We can only guess at the signifi cance of these findings for clini

calheart disease . They would appear to furnish a m ore realistic basis

for explainingthe consequences of ventricular arrhythm ias than has

been previously available . especially when com pounded by tachycardia.

O ne is also im pelled to s ender Whether certain degrees of asynchronous

m yocardial contraction m ight contribute to the lim ited perform ance

characteristics observed in clinical heart disease .

Followingthe incidental observation that arterialpressure rose

when visceral vascular hypotension was inniced a system atic inquirywas form ulated. it s ee toned that intravascnlar hypotension in the cat

abdom inalviscera supplied by coeliac . superior m e senteric and inferior

m esenteric arteries prodnced prom pt and snbstantial increases in the

system ic arterial pressure and heart rate . in the m ajority of cases.

this effect was seen even with intact carotid sinus and aortic arch baro

receptive system s but was accentnated after vagotonmy and carotid sinus

denervation. lntravascnlar pancreatic hyp otension also produced elevations

T he im m ediate ly aforem entioned experim ents on the callicrein

system and the previous experim ents indicatingthe readine ss with which

arterial hypertension could be induced by stellate ganglion stim ulation

suggested the possibility that it m ight be worthwhile to study the output

ofurinary vasodilator substance during the hypertension so induced,

Surprisingly, a m arked diuresis seem -red° T he objectives of the ex

perim ent were then altered in order to exam ine m ore adequately the

details of this interestingphenom enono T he data from nine com plete

experim ents show that upon stim ulation of the isolated steliate ganglion

in the dogthere occurred an im m ediate diuresis which appeared to be

independent of arterialpressure » and that duringthe diuresis glom erular

filtration rate usually showed little change and total urinary solute exc re

tion increasedo Followingragotom yo the diuresis was significantly re

duced although the elevation of arterislpressure was at least the sam e

or” m ore frequently, greater than that obtained before vagotom yi Left

atrialpressure deeressed during stellete stim ulation both prior to and

followingvagotom y and in seves of the sine experim ents the diuresis was

a function of the fall in left atrial pressure T he data suggest that, al

though there m ay be receptors in the heart which upon adequate stim ula

tion can m odify urise flow as suggested by G etter asd eo o workers o the

adequate stim ulus does not appesr to be as increased left atrial pressure

Laboraton of Kidnez and E lectrolfle Metabolism

Four m ajor areas of research are beingpursued in the Lab

oratory of Kidney and E lectrolyte Metabolism . T hese include : 1 )studies of the m echanism s ofwater and electrolyte transport in bio

logical system e o 2) studie s of the altered physiology in experim ental

heart failure in anim also 3) studies oi the control of aldosterone excre

tion in dogs and 4 ) studies of a cardiotonic protein system in serum of

hypertensive patients .

A theomr relative to the m echanism of elaboration of both a

concentrated and dilute urine has been published by D rs . B erlinero

Lewinsky» Davidson and E demo in general it has been postulated that

tubule urine becom es dilute by virtue of active transport of N e t from

urine into renal interstitial tissue , if no vasopressin is secreted, the

tubule m em brane distal to the dilution site rem ains virtually im perm eable

to water so that the finalurine rem ains less concentrated than plasm ao

if vasopressin is present» the m em brane becom e s perm eable to e ater s

water is abstracted by virtue of the increased osm otic pressure of the

interstitial fluid thypertonic in consequence of the transport of Nat» out

of the urine into the interstitial fluid alluded to above) and hyp ertonic

urine is elaborated. Mahatenance oi the high interstitial osm otic pres

sure is provided by a vascular countercurrent system in the m edullaryarea of the hidneyo T he presence of an osm otic gradient (increasedosm otic pressure in the pertinent area of the kidney) has been confirm ed

by Lewinsky, Davidson and B erliner o Direct experim entalproof of the

theory is now beingattem pted° T he prediction that urine in the loop of

H enle would he dilute under all circum stances is probably correct inso

far as the ascendinglim b of the loop is concerned» but that urine con

centration at the tip of the loop is not hypotonic has been shown byG ottschalh , in the light oi this findingthe theory requires m odification

and it has been tentatively propos ed that only the ascending lim b of

H enle is im perm eable to water .

in order to provide inform ation conc erningthe solute and water

m ovem ents involved in this m echanism , m easurem ents of osm otic

pressure » electrolyte contento and urea concentration of tubule urine

in the loop and elsewhere are planned° A nalysis of m edullary blood is

also to be attem pted , Dr o K ennedy is in the process of perfectingthe

m icropuncture technique for use in sm all anim als: ham ster, rat. and

nocturus . Drs tJaenihe and B ray in collaboration with Dr e B owm an of

- 29

D rs . O rloff and Burghave been exam ining the effects of various

agents on electrolyte excretion in the chicken. T he chicken pos se sses

a renal portal circulation so that substances injected into the legvein

pet-fuse the peritubular area of the kidney on that side prior to entering

the general circulation. By com paring the effects of a test substanc e

on urine com position of the injected side with that of the contralateral

control. it is possible to determ ine both those alterations due to direct

tubular effects of the substance on transport and the probable site of

action of the test substance . Usingthis technique it has been shown

1 ) that K r is secreted in the chicken. 2) that it is transported by a system

which is capable of saturation. 3) that it is subject to com petitive inhibition

by H e ions . T he dem onstration of a tubular m axim um for [G is the first

such dem onstration for a strong electrolyte in any species . Studies by

D rs . O rloii"

and Davidson are in press .

D re . O rlot'

i’

and Burghave also studied the effect of strophanthidin.

a cardiac aglycone. on tubular transport of electrolytes . T hey have shown

that the drug is a powerful diuretic . increasing sodium excretion m arkedly.

Changes in both K ai and H e excretion are consistent with the hypothesis

that the aglycone interferes with the tubular transport system by which

K -r and li-l ions are secreted into urine in eachange for reabsorbed N ae .

A uniform depression in the secretion oi”the anion. param inohippurate .

has also been observed. T he electrolyte effects are thought to be analo

goe s to those observed in the red cell. skeletalm uscle . cardiac m uscle ,

and kidney slices isee below) . in collaboration with D r . S. H aida. D rs .

O rloi‘

i‘

and Burghave dem onstrated 1 ) that the strophantbidin is secreted

by the tubule cells. and 2?that it probably acts on the contralum inalborder

of the tubule cell.

its effects of salto active adrenal steroids includingaldosterone

have been observed in the chicken kidney in acute studies . V asopressin

on the other hand dim inishes e ster excretion and prom otes the elabora

tion of a hypertonic urine . Vasopressin. an easily filterable polypeptide.

m ay act on the contralum inal border of the tubule cell since a unilateral

effect of sm all doses has been observed.

T he m echanism ofam m onia excretion continues to be a prim aryinterest of this laboratory.

"Urinary adaptation"(increased am m onia

excretion at the som e urine pillin the rat is due to an increase in th e

activity ofglutam inase . the enzym e which acc elerates the deam idation

of glutam ine . T his is clearly not the basis for adaptation in the dog.

N o difference in the activities oi any~oi the renal enzym es knosn to be

capable of providing am m onia from am ino acids we re observ ed when

either renal cortical slice s or hom ogenates of renal tissue from

acidotic and alkalotic dogs were exam ined (Dre . R ector and Orloff) .

It has not been possible to determ ine whether the observed effects of

acidosis are due to an increase in m em brane perm eability and influx

of precursor am ino acids . O f interest, howev er, is the observation

that the pattern of am m onia eucretion in"adapted dogs following the

infusion of either glutam ine or alanine is sim ilar to that of alkalotic

or norm al anim als . N o acceleration of conversion of the am ino acid

to am m onia was observed. in the rat the urinary response to alanine

was unlike that of the dog. H owever , definitive conclusions have not

been arrived at. Further studie s are in progress .

D r . C otiers has developed a precise , reproducible , electro

m etric m ethod for m easurem ent of true tissue chloride . lie is now

engaged in a system atic study of the distribution of ions in various

tissues . T he results are still in a prelim inary form .

D re . Burgand O rloi’

t’

have exam ined the kinetic s of N a+ and K 4

exchange in slices of rabbit renal corten. T hey have perform ed a large

series of studies in an attem pt to develop a reproducible and accurate

m ethod for m easuring electrolyte flunes in single slices of rabbit renal

corten. T he m ethod finally adopted is theoretically superior to others

in the literature and should provide m uch useful inform ation. T hey have

been able to estim ate K 4? inilun by m easuringthe uptake of K 4 2 by a

single slice in a s ello scintillation counter from m edium of constant

specific activity. E ffluu has been determ ined by the reverse process .

the washout of K “ from a single slice into m edium tree of iso’

m pe .

Strophanthidin has been shown to interfere with K “ influx W ithout apprec

c iably affectingK“ efflun. Studies of the effect of other drugs are in

progress . it has also been shown that the influx; of potassium increases

with increasingm edium potassium until saturation is achieved. Studie s

on sodium flux have been unsatisfactory to date . H owever, the negativ e

results are oi“

interest sinc e it is apparent that conclusions relative to

absence of linked N ae-K exchange in cortical slices based on sim ilar

findings of others need not be valid.

D rs . Burgand Orlol’

i‘

have exam ined the effects of strorihanthidin

on the uptake of P A H , N ae and its» transport, and 0 2 consum ption in renal

cortical slice s . T hey have shonn that the drugdecreases the K + content

of kidney slices and increases that of lino without appreciably affecting0 2 consum ption. A n observed interference with param inohippurate

accum ulation has been shown to be secondary to the effect on K + influx .

anim als with experim ental heart failure . T hey have com pared acto

m yoam tram norm aland failinghearts Na differeace in the yield of

acmm yosm its aedim emation pattern, viacaeity at reapcmse to A T P

has been noted Sfiudiea m Myasm A are continuing A n in put-tam

prelim inary finding 1 3 the fact that the m olecular W eight of Myoam A

from cardiac m uacle appem's m I‘m the w ane m that from M ental

mammals» ° This m ewww m mner repor mm the lifiem ture , N o clearcut differenceam the ifim mmc tenmm patterns Qfi

‘

glycergl extracted

fibres from film two gm anpa of hearts have been obm w ed o Studies on

water and NM ewww mi cardiac m mmcle from hypaphysectom med m i

m als are in pm g mgao

Laboratory of T echnical D ev elogm em

T he Laboratory“s developm em ofnew and m ore somsifi ve

m ethods and instrum om o ia comtim zixngalong severalMo oo o T he

deve lopm ent of m ethods for analysis of m aterials obtained by m icro

puncture techniqueo has shown oom e prom ise in the form of a m ethod

of analysio of the olkoli m etals by a. rather oim ple o rapid m othodo

T his m ethod otiliz oo the plasm a glow of a. radio frequency dioohargein helium at atm ospheric prom o t e to volatiliz e m o excite tho alkali

m etals from a platinum W firo o T he om iooioxm oo producod is separated

from the helium Magoo by m eans of a m omoohz om otor and m oaoured bya photom ultiplier om ! im ogrotimggawonom otoxn Sam ples are depo sited

directly on the plotimum wire from the m icro o

pipotte and to date i?

appears that m oles of oodiom can be ootim ofiod with an accuracy

of 592° Much grootor oommfi vity oppooro ofitamoblo but this m othod has

owly recently been oubjoofiod to ioteooivo study . it appears to have We

vantage over proviouo oppmoaohoo ,

F ree z ingpoint dotorm fimofi ooo om m icropwmowro oom plos have

be en; m ade by oboow ottioo of the tim e ofm owing A dequate m easure w

m em o cam be m ade tam ing oom ploo m we w age of oovorol thoo om d mo

of a. cubic m illfim otom butwe m om od mm m omm a further ovoloatiom o

T he dovelopm om of a new technique ofi extrem e om oitivfity for

goo chrom atogm pmo m oiyofio boo boon com puted and oevorol m odifi

oofiiooo off"

oxiofimg ohrom ofizogm pmo toohmoo oo hw o boon m ade o T he

developm ent of tho high oomofitivfitg? dofioofior hag m ode if; poooible to do

om m olyoio on a few m icrogoom o of fatty acid m omyfi ootom go that

m ethods fox? handlingm icrogram om ploo head to be developedo So c:

o ooofi’

ol oom pko wtxooootioga oyofiom o oopoblo of?accurately and m oron»

do cibiy mgoctfio ga few m icrogrom o of m otormlo mm heated entries to

tho colum n homo boom dovolopoo and o dooompfitmn m om m a

During the oum m ox" the nuclear m ogmofiio room m oo flow m otor

project woo m oofiivatod and o oorieo ofwow inmootod that mm took»

mquo (zoom m omm y ho mood to m oooux'

o flow in o reprodooiblo momm a

T he ooooooifiy for additional emomm c controlbecom e ovioom and a

m ore verootilo appeal-ammo mow matador om ofiruotiom io mooring com plefiiom

T ho oomotm otioo of this oqmpm om m boringm ode poooiblo by our acouioi

tiom ofi”Man, V oovokoo K odrm voov

fio oom fiooo o it is hoped that mm oppom m g

willm at ©3313; allow am to ovomoto tho oppfiioofi om of nuc lear m agnetic

rooooamoo to flow m ooom'

em om best will, aloo provide won-king equipm ent

to 33 5 115the semaitivity ofbrea d line m m mnm ce m ethade an; analytical

technique o T he advantage O f this technique lies in me beinga m ama

destructiv e m ethod as? am lyais m Oppoafid m m having a high m agis

A m gkectm nic curve m alyz er em pable of ay mkaefi izmga. cam

pcbsite curv e 03“

aevem l pure fm ctigma hm been, com plefced o T he

0 &m t Manama 5m m w um gamma curve m pm fium ed to indicate

the individual fum timma m umm im d in mm cmnplex «aux-we e T he; applica

a

mamaQfmm appm am a m we m allylw emfm um s?!G amaaiem diatribufiians

m expected m facilitate the m lyam O f m any typefi Qfm gical data

norm ally awafi able m mm farm o T he pm pm atimnof pure sam ples Em

fiemmgthe apparatug«m an t fical prablfim im mwmg the rem lutmm caf

Qvemappmxxg abm rptwm cwmve g m m psagrem o

Sam m m fmem emm film the: m im analyssm appm afma have be en.

m flde m the fi’

m 'm Of ehim im xtimgthe awed far a pm e ram m analyz er m

c®m firm ma attainm cem m?the mall pmm o A wqum m law ‘

m ntm em r hm

been 5 1m m m be a m fi afacwry m m deteatm o

T he m am waive catheter tip trm gducex has bean ghawm m be

feaamle but. tecm galpm fifi em s m gm ammcfaigm m’

{gm tip a nd adequatem am atrm sfer dawn the m am m mm mem m m be gamm a A gam e

interfem m em r 11533111312n @@m @ m fi‘

med eiectm m e zecm iquw m imcm w e

the aem itivmy Ofm em e m am wm hm M aym m m fi pm m iae of bm fim ing

a n©mm de 5 tm ctiva m m pm pgmy demicg whieh wmam auraw predmmm e

hehwim m 3 gm chrm zm mgm phy datm m r o

T he pr©fimcti©m m?fn hydmmxyh m dfixm m by mm fim mm eavimngam

hag beam review d mind, ©©m paw d m the pw dm tmgnQfi’

free radicalgs bymm zmg radiamm o

h am m em fimmm my anallygig a)?flucam m em mm phwsphfim am m

m ateriam m g lbwm QQM‘

tfimued ai mingfilm Maw Qfi’

m tfiiamgup a m efiwd

fm ‘

ammmgmad Eater m afiyzmgfirm em itficfid phw phw em em flpectm m

pmaducefd by a h igh intenmfay lh© 1m durafi mm mm aam m e o A 9m m »

eamdm mv e: zinc: 0mm w fi am i555 glm fimifim and mngwmg expm ure m

the m m @pectm m that 5 113??a is"m ad am

"by m aem a m?a vibrating,

reed 9m m ©r m eagum d aza tam cargfim pxmfim ed by «iim harga @f the

pap er by a m gvmgbgm a)? light 18mm m am m t"aygmm g news he

fluccegamkhm fih© m ammal} mmmgwith am msmmxhag mat

liver have been m easured and ueed m cem pute the specifi c eurfae e

per gram dry weight efi‘

the fro z en m aterialo T he efficieney of varieue

cements for rapid fireseing has been m ee euredo Kt bee been found that

eppre zzfim etiem ef hem tree efi'

ee at the surface ie given by

9 where T 5 is the surface tem peg'me re o Tgie the fiem perawre

ef the cement» and E , which ie new ly eee efiem » v erie e from 0 . 2 sale !cm z eee o dege fe z,“ liquid pregam e te o m e ele /em 2 eee o deg; fer liquidmitm gexno K H is n ew fe z e cement it ie peeeible fie develep an em u;

weary fer the k eeeling efi“

e eelid eff regular eeeee wifiheut phaea ehm ge

end an eppmemlm ete meek-

y fe r eeelmgwith phe ee eheege o By epplieetien

of the theery fie experim ental ceelmg curvee it w e be determ ined W hether

e eee ee eheege bee eeeum ed c Se en em; ee elyeie efi’

eeeling ee ee ee of

ge latin bleeke be e me mefted that fe e w hee leee them; 1 m m em edge » eeelee

in liquid emeem e m ere ie ee ly pertfiel {free z ing efi‘

the e aten Week has

commuted em the peepeeetiee efi’

epeeim ene fer eleearee m icre ecepy by

free z ing and drymgo H ere , in order to get e euitebly e e iferm m ete »

ple em ie m eterie e e e fleet ebjee’

zta eelemaiee ex“

been the m e eee C haee

C hee e w e eifpe rm eefle have been eteetedo

A em dy knee been e eeem ekee efi?the em ery efi‘

tree epert in linear

hiexegie el egret-ew e , typified by eteedy etete fireeezr emdiee p and e m eppiim

eemiem: ef thine enem y fie epecfifie peeielem e m the kim etie e ex?fatty acid

m etebeliem . T he beefie theery relefiee flexee in e, eyetem by en integral

eqe eeiee e? the type : ”3m 3aft

{as } e “ a w ) du g

Where e m fie e tee nepem fim emee emereefiemefi e ef the eyetem o fie m e le e

mm preelem e a the flexee ewe eeleeee by e. eyetem eff eeeh integral ewe »

tim e , G ee ex’ ei m eteie; m ethede fee the eemtiee efi’

eyetem e ofmm kind

have been dee eiepedo

"m m thee ey be e been enem a the give m e wello km ewe

eele tiene fer eyetem e whim eemeiet effeee em l e e é fi’

erm ly m ixed cem e

pemm em eo end knee beefla «applied fie geneeel peeblem e eff chem m elyeie

end m eeel eee etm efiiee m eyetem e with meem plete inferm efi emo m em u»

leberetien with D an B enem F eedriekeee s the m ee t }? he e been applied to

the m elyeie efi’ teeeee defile en m ay eeie m etebeliem p with. the prim e r}?

ebjeetive efi’

eem perieg the m etebelie pefl m eye efm eeterifi ed fetty eeid

(UF A Dend ehylem fieree triglyceride fatty ecm QT G F A EO Sem e enema

hee d eem pmafi eee hewe been «se emed em, mad the prem em hee been per»

tielfiy program m ed fer en elee ‘lreme eem petee o

Em edcfitfien te the regime pmejeeme , fine Leberefitery ne e previded

epeee g eelleberefi ee and eee eeltefi ee with the C hancel and P refe eeiem l

E de e etiee G reepa headed by D e Mem ey B ree ze» fie ee elee fie the m ug;ei

’

eeler telewiefiee teeh mquee m bielegieenreeem eh o

Laboratory a? Chem ical P harm acomm

B im bem iafig of Functmm

Much @f the current wm k m thia labm awry is related to our

pbiwsophy that the gap betwe en phygmlow and pharm acm©w am one

aide and biochem istry on the auger atem a largely mom the agsum ptmn

that Specific organic fm ctiom can be explained in term a of the"umi~

vereal"reactim s a?wterm ediary m etabalmm o Envemigatim win this

laboram ry are based can the beingthat gpecialim d fumcsim s are clemelylinked m u nique em anates , age far exam ple » peeriphe zm lmem e sand man

tiara 1 8 © 1033e w am iated Wm » the aubatrmes acetyiehglime and mm »

adrenaline (N E ) .

Role 0? M d. mm

Moat drugs which infl uenc e the peripheralnervous sygtem act

at jumctmna Where nerve im pulaeg are mm gm imed by aegtylchglme 0 1

N E , B37 m amw (3n Magmaaffem fl m cm fim lmmrvmw aystem (C N S)by m adifi

’

y mgchem ical trm gm mgicm at am ague jw ctmmg o E vidence

m mm is diffi cult m Qbmm bm m me C N S stm amm a are inaecem ible

and becam e aeefiylcflmlime hag beerm the (M 13? eatamwhed centralm am a

hm 'm m wo T he caim m mry 0 5 m e mm vem ma xim um in brain; (pfm e

imtemeely native am fimw g N E mad m m mmm WT ) » m akea it ream nm hle

m am um a mm them m bgfim cea hm m apecialfim d {m etim za 311m 8m m

Mam e they are figm de

Tug w mcepticm a gfiW e R a E m a have hem awed aa a fram emmrk

mm bringingwgem er M echa nical» phygmlogicai m mphaxm acowgmmaapem s afbrain fiw ctwmo flew infl amed that W Q Qppw img sym em aa

erggtm pic mad trvpiw fmmpic o integwam mam z mm ic with} aom amm mm ‘

and psychic mm fiiemg 32mbram amm o T he m gmtm pic «im am integrates

aym pafihefi c with m m a‘mm aam r m aivm es m pm dum the behavwm l pat

»

term of pm mwe actim o T he m mm fl afim m of: ergmm pfic pm m m im me

are arm-

mails im m am d aym patbefiic amivityg enhanc ed m uscie make and

am activated paycmc amm o Hm com ram the txm hm rapic divisiom integgatee

pam aym pam etic with m m am mr m fi vfitfieg m prgdume hfihavmm l pat»

terns » xecupermwe m mafiure o T he; m verfiuwfi’ecm emf tm phmrmxm pram

d©m mw ee are drw mfineaa m mgfleep » im m aged pm m ym pathetic activity

da m m ed akemtal tmae m aaetifi tyg m esalmm m d m gzmm e w external

Chlorprom ssine and other phenothis z ioes have been conclusive lyshown to decrease sym pathetic activity by 5 . control action and have littleor no direct peripheral action. it is pertinent that the pressor effectsof cocaine and chlorprom ssizie are com petitive , suggestingthat theycom pete for the som e receptors .

Me s s H T.M M

C om pounds like reserpine elicit soostioh , hypotension and a

vsriety of other eitects like those of chlorprom ssioe . it is believedthot they do not sotas} ; 5 3 hot by liherstihgm ines through on actionon the m echanism s thot coocehtrste the m ines in body tissue s . R eser o

sine appears to stim siste the troohotropic system through the actios of

free h it is consideriogreserpioets mode of soties it m ust be kept in

m ind that it im psiss the storsge of N E sod ii’

i‘

but not their form ation.

When stores srs desistco, syothesis of tho fires sm ises continues. it

form ed slowly, the w ise m os be com pletely m etsholised by m ososm io e

oxiosss (M5 0 35 355 the leve l st seceptoss he too low to elicit a response .

Under these conditions reserpine m sy he considered to have depletedsynoptic junction ot

‘

hom ooc, Cootrsstiogly, if the m ine is form edrsoioly, the les si st receptors m oy he sufficient to procioce a persistentrespooso ,

studies on tho tosoos es oi hosts h’

i‘moleste thst it has 5 rapid

turnover, 50 per cost is 1 0 to 1 5 m inutes, com pared to several hoursfor NE , Thins, otter resei

’

pi'

oe 5 5 5 5 5 5 5 5 5 5 5 , hrsio H T m ay he form e drapidly enough to produce 5 tsoohotsooic rsspoose . These results are

coosisteot with the thesis thot fires it'll“ is the im postsst tector in controlreserpine settos ,

T he ges sosi ooioioo that the seostive soo hwotessive actions of

reserpine hos e 5 com m oo csotrsl stochm ism m ost 5 5 5 be m odified . in

low doses , rsseroioe ooplstes oesiohersl N E;without affectingbrain H Tor N E sod s ithost prosochig 5 5 55 5 . A s 5 result, tooctiom l sym pos

thetic oeoew stioo is oroooceo. A com plete soossstioo of peripheral andcestrsl offsets is 5 5 5 5 with 50 31 1 5 tC ihsb, 5 sem i- synthetic m alogofreserpine , is sotoli 5 55137doses , this om gosolstes pe ripheral N E without loweringhrsio sm ioes , 5 5 5 5 5 are not seostso hot show hrsdycoroto , hypoteosioo , oiM oisheo response to csrotid occlusion and othereffects of chem icsl sm oothectom .

N E depletion, however, does not account tor central eti’ects oi

reserpine . This has been shown with com pounds that affect N E bindingin preference to that oi l F or enam ple , sm all doses of St] 5 1 71

(C lient. a reserpine m ales, depletes brain of N E but not of H T , and

anim als are not sedated. h owever, higher doses lon er brain MT and

also produce sedation.

Pharm acolopic evidence that reserpine does not act centrallyby NE depletion was obtained from ohsernations that, aniline chlors

prom asine , the drag does not decrease central sm athetic output, butincreases parasym pathetic ontpnt Than re serpine e licits m iosis , lecrie

m otion. nasal secretion and enhm cem ent oi light relies by central parasym pathetic action, T he decreased sym pathetic activity, e . g, decreasein blood pressnre and relaxation ofthe nictitatingm em brane , is explainedby peripheral depletion oi N E .

A dram atic difference hotn ess reserpine and chlorprom asine is

that reserpine elicits tonic close rs at eye lids and extrem e m issis as in

norm alsleep, While chlorprofm asine prodnces slight m issis and releas e

tion of eyelids tptosisl. This.

accords with the notion that reserpine m ay

canoe sedation hp stim nlation oi a“sleep

" center and chlorprom azineby snppression of an aronsalm echanism .

A nnm her oi com ponnds that inhibit m 0 are need in the treatm ent of depressed m ental states, hyp ertension and angina pectoris .

lproniasid, {in his, «Eh his andJE set , all oi which elevate brain levelsof ill?and, in certain species, llh , have heen stooled

,in this laboratory.JE sis and EB 835 are estraordinm wpotent agents o n about 50 tim es

m ore so than iproniasid E vidence sassests that the central eacitationproduced hp these com passes is re lated tem porally to the rise in brainN E rather than hrain WT . Fartherm ore MA O it itors elevate brainh

‘l‘

hnt not NE in cats and dogs . These anim als are not escited by the

The elevation in hrain N E prodnced hp the inhihitors indicatesthat MA O inliills an im portant role in the etiologic inactivation of this

am ine . O n m ethpltransi‘

erase does not seem to he involved in the inactivation oi brain NE . in tact, direct evidence indicatesthat brainN E ism etaholised alm ost entirely hp seas , There is som e evidence that the

m ain role of MA O is not the m etabeliem of N E releaeed at nerve

endings but the regulation of the em euh te ef N E and H T within the

h em-0 h so that they do that continuously 3pm onto receptor sites .

A ntieehvuleeh t P regertiee of Mah eem ih e O etde ee Ehhibitore

MA O thhtbtten' e elevate heath em th ee m aM e ek electreehe ek

eehvuleieh e 11h rete eh e m etre z el eeh vuieteh e in m ice » in contrast tere serpine whteh leweee the beam em th ee end ehheh e ee the seizures ,

Strehg evidence euggeete that the effects et’

reeerpth e and of MA O inte

hibttere ere m edteted theeegh the ehehge ih beetle em th ee ; 1 ) Afterthe adm inttetretteh et

‘

a MA O thhfihtter the ehtteehw leeht effect iscleeely related te the etee ht em th e lee ele; at hitter reeet

’

pth e the ema

heh eem eht of eehw leteh e te re lated te the deetih e th am th e levele; Sbrelee ee at em th ee by reeet

‘

pth e a hetee e treetm eh t with itpeeh te etd0

eliette the ehhehe em eh t et’

teet et’

reeerpth e ; t) tpe eh iaeid tellewed by

giving reeerpth e eh eite eh ty the eh tieeh ve teeh t ett’

eet et the MA C te

hibtter . H eweve zro evideh ee that etthem N E QB? H T te the tm perteh t

teeter te eeutveeet; he hatht em ih e eelee eed by reserpine end the

m eteheliem ofwhich te hleehed by MA G thhthttezre m ey he ih voivedo

Since the hleehe tte eff MA G eepz'

eeeh te e h ew type et’

m eeheh iem

th the eeppt'

eeeieh et’

experim ented CQQn iQttag e. elth te el trial ofthedrege te ht pregreee ih eetteheeetteh with th e Shy ,

N E er epth epheth e ht deeee et’

0 ° 5 te 1 m gper kg» imfueed evera peried effeeveeelhem e » th e me e revee ethte tetty th fi

’

iltxetieh 1h heartend ethet' tteetmee o eeeeeteted with e reverethte hypereeh ttttvity ef

’

heartte epth ephrth e

c th eh e ed eeehythm te e o T he hptd eheh gee are bleeked bydiheh eylth e o T he ee eteete heme he eh exteh dee te the effects at 0 0 1 4 9eleehet m e ethtetmth e o Eh eth gte ee eee theee ettheteh e ee eeuee e tt iglyceride

M tltretieh he met h e ere g Whteh te hteehed hy the irreverethle edeenergie

hle ehthg egehte o etheh eyh h e o diheh em th e hate the revef ethle egehto ergeta

em th e ,

T he ee eth ete l ere effpetehttel tm peeteh ee cheese 1 ) e etechelem ih eem ay he th vetved th tet treh epet tg 2» the petm emf etep ht th ug liver dam age

m ey t welve thtert’

eeeh e e with e, eeteehetem th e m eehehtem that regulates

the lipid solubility of the unionized m oiety, and the pH at the absorbingeite o Several studies have been concerned with further elaboration off

thin cone ept°

Intestine

T he steady etate diatribution ration oi”

druga between plaam a

and em ail intentine are predictable it”

it in aeaum ed 1 ) that the intestinalm em brane aeoretee hydrogen tone that m aintain a z one at its earface

at a" virtual" pit of 5 ° 39 and 2» that foreign com pounde are absorbed

large ly in unionized forth and at ratee proportional to lipid solubilitySim ilar re eulte were obtained for the co lon uninga

" virtual" pH of about6 0 5 ° T haeo reeulta explain the poor abnorption oi m any water solubledrape .

B loode B rain Barrier

F oreign com pouncle penetrate into the CN S alth oet exactly an

predicted on the banie of their phyaicochem ical properties and the

aeeuniptione that llonly the unionized term croeeee and 2) thin m oietyentere at a rate proportional to lipid lolnhilityo

P re lim inary reeulta indicate that barbital paaoee slowly frombloodetreani into hraino thene e into C SF O T hin ouggeote that the choroid

pleirue in not im portant in the paaaage oi’

drape into the C SF ,

C ertain foreign com pounde have been tound to penetrate rapidlythe pituitary gland» pineal glan d and the area poatrem ag Whereas theyelon ly penetrate the brain no a Whole T hin findingnot only euggeeta

that theee arena are not part of the brain hot that like the pituitary theym ay aleo have aeeretory {function T he obaeruation with the pituitarym ay explain ite aeneitirity to epinephrine » deeptte the poor penetrationof the am ine into hrain o

B ile e B lood Barrier

P revioue n ork diocloeed that inulin and eucroee » large lipide

ineoluble eubetanceaa appear in hile in aignit’

icant am ounte o indicatingporoaity at eom e local in the hepato e hiliary eyotern ° F urther atudieehave included a annaller m olecnle o m annitolo analindicate that em ailm oleculee paea the ”

bloooc h'

lle harrier" m ore readily than largem oleculee o Mannitol occnpiea a volum e of dietrtbution appromm ating

totalwater of liver; inulin and sucrose occupy a sm aller spac e but

still one that is probably greater than extracellular fluid , P relim l

nary results eugfigest that lipido eeluble fereign chm pm mds do not appear

in bile in appreciable am chh te but that lipida ih eelhble m etabelite s and

quaternary com pehh ds m ay appear in high cencentratieh o

P e saa e of 0 mm .

A cm es M em branes

T he View ci’

a lipeidalbew eary between iateetih e and plaem a

perm ite inferencee ae to whether a eahetah ce ia ahaerbed paseivelyer by a apecialiaed preceee o T he ih teetihal aheeeptieh at thym ine and

its eccleeaide thym idih e ihvelvee a taah epe i't m echah iom o T hus , the

degree ei’

their ahee i'ptieh decreaaee with increaeihg ceh cehtratieh o

Uracil depeeesee the aheeaptieh at?thym ih e «he m ethyluracilbby com peting

for the m echah iem o P relim ih aey reaalte eaggeet that a hammer at purine

ah apyrim idine derivativea cem pete fe i the aew e trah epcx't m echah iem o

hi centraetg high ceaceh taatieh s ei’

D o

glace ee er Le histldih e do eat th u

hibit the trah epcrt at thym ih e °

T he m etaheliem at damage ie eliacaeaeci ih teem e at”

theee cate e

gerica at auh etah cee

m biinterm ediary m etahciiam

F f v ved

T hie lahe i-atci'y hae extended ite eth diea effairingm etaboliam to

the ee e called ah tiih etahciitee g ehhetah cee atm cte rally ee eim ilar te

h erm al eahetratee that they becam e iaveivee in the”‘h tiivereawenzym es

ei’

ih term ediary m etaheiiam a T heee cam peh h ee are h eed ae antle cah c ei'

ageh te and their m etahcliem ah am echah iaih ct?actieh are cle eely tatera

twih edo 66° CLP E>hae heeh tchhate he a cem petitive the

hibitei" at ph eih e ahahypezzah thih e eatdatieh by aah thih e eatdaee o it is

ceh e eeteclte Gw chlereh i-te acid «Se CLUh a hereteteee ahhh ee m com pound

which hae heeh caye‘

talliaed ahachaeactex'iaedo T W Q ether peeducta are

alee term ed in viva ih cleclihgm ic acie‘

t 6° CLP allec ehtere iate hyac

thetic reactieh e ah aie ih ceEpe i'

ated ihte hacieie acido E xperim eate under

way will aeceetm it any ho C LP m etahelite e acceah t fer ite ahtie cah cer

eii’

eete °

- 4 7 ,

(2B Substance e acted b rela lytive v'

non c

m c fi ic ema mw ie i vo ve

A lcohol and aldehyde dehydrogenaee e are exam ples of extrem elynono epecii

’

ic enzym e s . T he norm al role oi”

aldehyde dehydrogenase but

not of alcohol dehydrogenaee is known A lcohol dehydrogenase is in»

volved in the m etabolism oi“

drugs from which an alcohol is form ed as

the iiret step in their biotraneiorm ationa e . g, barbiturates , Studienare beingundertaken to investigate the possibility that variation in

hum an tolerance to alcohol m ay be bae ed on difference s in alcohol de w

hydrogenase activity P reparatory to investigatingthe biologic varia

tion of the enzym e activity in a large populations the enbatrate Specificityoi the enzym e is being etndiedo Snrprieingiyp higher alcohols have a

greater affinity for the enzym e than ethanoio it in possible that other

alcohols m ay prove to inhibit ethanol onidation and time provide an inter a

eating experim ental toolo

M W e “ an iv “ Qtv d

Moot drag enaym ee fall into thin category F urther studie s on

the oxidative enzym es in live r m icroeom ee have been m ade

(at T ine enzym e that onidiaee ctaiorprom aaine and other sulfurcom pounds to the correeponding enii

’

onide derivative has been atudiedo

it too requiree T P N ii and 0 2°

to» T he m icroaom al enzym e that onidiae e hydrocarbon side »

chains to alconoia also requires T P N H and 0 2° T ina ie an im portant

enzym e in m aintainingbody nom eoetaaie einc e food centaine m any

terpenee which cannot be excreted onieee oxidiz ed,

to» T he key reaction in the m etabolieni of nicotine occurs inm icroeoniee and reoniree T P N i

—i and (9 3° it involvee oxidation of the

carbon atom neat to the N in the aidecnaino T he next etep to etndy

presum ably invoivee ring emitting» a reaction not yet elucidated fordruge o

(db Monoam ine onidaee inhibitore aii eeem to antagoniz e a

num ber oi“

m icroeonal enaynxee o Tnia pecniiar action in unrelated to

inhibition oi MA O ; in fact it in reveraibie and dieappeare by the tim e

that MA O ia com pletely blo cked ,

then a powerful tool for studying the m echanism and the sequenc e of

biochem ical evolution is available . F or exam ple s toads oxidise drugs

by dehydrogenation, in contrast to reptiles which hav e system s which

ahtivate oxygen , But m eadow frogs , which can live on dry land for 4 8

hours» m etaboliz e drugs by m icrosom al system s requiring T P N H and

0 2, T his suggests that the m icrosem al system s originated in am phibia

and not in reptiles ,

T oad and salam ander tadpoles cannot m etabolise foreign com e

pounds until they have undergone m etam orphosis O i interest is the

findingthat guinea pigs and m ice are born without the ability to hydroxy»

late barbiturates a dealkylate alkylam ines o reduc e nitre com pounds or

term glucurenides o T he appropriate enzym es begin to appear in the

first week after birth

F ish can xterm neither ethereal sulfates nor glucurenides , H owe»

ever g these reactions are present in am phihia and in higher v ertebrates

A considerable pussie s however » is the finding that though i’

ish m icro -a

som es have the glucurenide transierase they lack the enzym e required

for form ation oi active"

glucuronide ,

-

uww.

A ce tic“ A cid and a New P awnww ofm acaw

M etaboli sm

«a» Studies on hie synthesis oi Lw ascerhic acid which suggests ,

a new pathway of glucose m etabolism have previously be en reported an

iollows z D e

glucose D e

giucurenic acidm m _ >

L-

gulenic acid L o nylnlese 7D o xylulose_

pentose cycle (30 2° D efinitive evidence has now be en th e

tained for the occurrence oi these reactions in the anim al, T his schem e »

referred to as the glucurenic acid pathway , is im portant for severe

reasons : iii L a

gnlenic acid is converted to L=ascerhic acid in rat: and

other anim als which can eaist without dietary v itam in, whereas m ans

m onkeys and guinea pig lash this enzym atic step ; «2» this pathway GK “

plains the origin of La nylule se o the sugar excreted by patients wits

essential pentesuria; tn inesitel is m etaboliz ed to an appreciable extent

via this pathway sinc e it is converted to c lncurenic acid and L -

gulonic

acid; ti) this pathway m ay he involved in the synthesis of D o ribose present

in nucleic acids .

T here is, at present, disagreem ent as to which pathway of

norepinephrine m etabolism is of m ajor im portance in vivo . T he two

dem onstrated pathways, MA O and O n m ethylation, are under study.

Methods for assay of com pounds form ed by the latter route are in

process of developm ent. T echniques for inhibiting the in vivo m eishe

lism of norepinephrine by adm inistration of com pounds which block

either MA O or O o m ethylation. or both, are under study. It appears

that only when both pathways are blocked can the rate ofm etabolism

of norepinephrine be sim ii’

icantly decreased.

T o o am ino acids which are found only in collagen. hydroxy

proline and hydroxylysine . hav e been under study for som e tim e . T he

inform ation from these investigations willbe valuable in studie s on con

nective tissue form ation and pathology. A pparently hydroxyproline can

be incorporated directly into collagen in rapidly growingtissues such

as chick em bryos . Conversion of proline to hydroayproline has also

be en dem onstrated in cellfree system s. K etoproline . the dehydrogenated

analogue of hydronyproline , has been sheen to increase tissue levels of

hydroxyproline in anim als in vino . it can do this because it blocks m etabo

lism oi hydroayproiine so it is directly converted to hydroxy

proline by som e ensym as yet unknown. This com pound will

be further investigated in vivo to see if it can influence collagen m etabolism .

T he product of the action of glotam ic acid decarbonylase on its

substrate is ywaminobutyric acid (G ibbs ) . T his am ino acid is found in

highest concentration in brain and its m etabolism in m am m alian brain

is therefore of great interest. One roots of m etabolism involves trans

am idination to yield 3;~wanidinobntyric acid. T his com pound is also

found and form ed in brain. Like G A B A it produces inhibition of central

cortical synaptic activity. The significance of y-guanidinobntyric acid

and its further m etabolism are under study.

Other routes of GA B A m etabolism are also of current intere st.

incorporation into analogues ofpm tothenic acid containingG A B A instead

of s-alanine have been tinder study. Hydroaplation of G A B A tofl-hydroxyG A B A has been reported. attem pts to corroborate this and study its m ocha

nilxn are in progress.

Studies on phosphorus and calcium m etabolism were carried

out in ll patients with hyperparathyroidism before and after surgery,

and contro l studies in 6 norm al subjects . A n estim ate was m ade of

the relative value ot‘

phosphate c learance . response to phosphate with

drewel. and calcium infusion as diagnostic tests . T he defect of calciumm etabolism in sarcoidosis was further investigated: the studies show

that patients with hypercalciuria and sareoidosis do not uniform ly show

sensitivity to vitam in D . increased absorption of calcium , or response

to steroid therapy. A ttem pts to produce radioactive vitam in D were

initiated to aid in this phase of the study.

The effect of hypercalcem ia and hypercalciuria on the renal conc entratingm ect m aud i os renal sodium conservation were investigated

in a num ber ofpatients before m d after m ofi fication of the hypercalciuriaby therapy. T he inability of these patients to reabsorb water was con

firm ed and found not to be related to an inability to reabsorb sodium . T he

presence of the defect correlated better with the urine calcium than with

the serum calcium or the therapeutic agent.

T he antidiuresis of cirrhosis was investigated and was found notto be related to antidiuretic horm one. but probably to depend on proxim alsodium reabsorptlon: it could be decreased by infusion ofm axinitol. and

reproduced with rigorous sodium deprivation in norm al subjects.

P atients with potassium - losing renal disease were studied with

m etabolic and renal function techniques. Whereas a num ber ofthe se

subjects appeared to have renal tubular acidosis as the underlying die

seese . others showed the disorder despite norm al ability to secrete

hydrogen ions . T he role of aldcsterone in these syndrom es is beingstudied with the use of all agents knee s to alter aldosterone secretion

in the norm al state .

in studies in rabbits and dogs the transfer rate of album in and

other large substitutes across arterial c ells was m easured by the intro»

duction of rafi oactire labels . The role of absolute pressure. pulse

pressure. and alteration of the endocrine status of the animals was testedin acute and chronic studies . Both radioalbumin and radiochole sterol

m oved out ofthe pronim slaorta much m ore rapidly than the distal sorts.

T he eilects ofbovine growth horm one in m an were studied in a

number of obese and normal shbiects to determ ine whether the fat contentof the diet would alter response to the horm one . it appears that this

inhibition in the gut and possibly liver, s. m ethod for m easuringoverall

MA O inhibition becam e desirable .

A sensitive and specific m ethod for m easuringurinary tryptahainewas devised. Marked increases in the excretion of this am ine were ob

served in patients treatedwithJB - 5 1 6. T he level of urinary tryptam ineappears to be an excellent index of the MA O inhibitory action of investi

gational drugs . O tnes am ines previously undetected in m am m als m aybe found in the urine of patients when MA O has been inhibited. We have

already positively or tentatively identifi ed tyram ine. o o tyram ine and

phenyletliylam ine under these circum stances .

Since it appeared that am ines other then norepinephrine and scre w

tonin m ight be involved in responses to MA O inhibitors. the pattern of

cardiovascular response to various am ines is being studied in the dog

and in m an before end after MA O blocksde . P otentistion of the response

to dopam ine and trwtam ine butnot to norepinepltrine and serotonin were

shown in the dog. A m erited potentistion oi”

dopam ine pressor actions

has been confirm ed in m en. Thus. it is conceivable that dopam ine inthe sym pathetic nervous system . m ay have a role other then sim ply servingas the precursor ofnorepinephrine .

(29 M ismetnbol_

A m inoacido in Men: The urinary excretionofbound hydroxyproline DPR W es fonnd to be elevated in 8 of 1 0 pstients

with Marfsnfls syndrom e. possibly reflectinga basic detect in the m etano

lism of collagen in this disorder . The dsily excretion of bound O P E is

independent of dietary intake of the m ine acid unless it is taken in theform of ge latin. Dietary alterations in O PR have not produced any altere

tions in patients with M ews which are not also shown in norm als. T he

influence of other am ino acids end proteins willbe studied further and an

attem pt m ade to study the body pool ofQP R with labelled proline and O P E .

T ryptophm loading in m en has been shown to increase tryptam ine

excretion en s field. When this s ee done in petients onJB c-dls, higher

levels of excretion were obtained and the patients com plained of a drunk

fe eling. A lthough. ingestion of tryptopnsn is nsnslly innocuous. under

MA O inhibition it appears to have definite pharm acologic effects . P reva

ions stones on indoleacetic acid «we.» have been extended with the fi ndingof increased excretion in several gsstrointestinsl disturbances other than

spree. inc ludingpancreatic insufficiency end blind intestinal leap . in »

creased M A secretion in these disorders is considered to be due to altered

this are beingfurther pursued. (2) A study of the e lectrical deform ityof the Wolff- P arkinson-White syndrom e has been com pleted and pub

lisbed, T he interestingfindingwas the high incidence of abnorm alventricular conduction in this syndrome » the characteristics ofwhich

shed light on the probable mechanism of the syndrom e . (3) C os sid

ereble effort has been expended to increase our supply of autopsycoistrolled E C G data for various age groups stud disease entities . We

have wellbeyond 1 , 000 case s new in this file , The analysis has con

centratedo so far . on definingnorm sl ranges of E CG data in infants and

children, and in the age group beyond 60 years . C ollaboration with D r ,

G eorge Manningof the R C A F will augm ent our data for the third and

fourth decades , T his will be the first statistically satisfactory analysis

that has ever been m ade of autopsy- controlled and age - controlled E CG

data.

is Open, however , the indicator im m ediately appears in arterialblood

and in high concentration. T his technique has been fully evaluated inthe experim ental anim al and is beingused in patients operated upon

for esOphageal varices . The indicators m ay also be used to determ ineand perhap is lquantii

’

y valvular regurgitation. in the course of cardiac

catheteriz ation the indicator is injected into a clim ber as a sam ple issim ultaneously drawn from the next m ost promm al cham ber . When

the valve is com petent the indicator is found promm ally only after nor

m al recirculation has occurred. But when regurgitation is present it

is im m ediately detected. T his study is a routine clinical applicationand attem pts are beingm ade to provide it with a quantitative basis .

T he Section on Cardiology has carried out a group of studies on

the physiologic effects of digitalis in patients . in norm al subjects , the

effects of rapid digitalisation on cardiac output and central blood vo lum e

were m easured. P relim inary results indicate that the work of the leftventric le and the cardiac output and central blood volum e are significantlydecreased. P atients with atrial fibrillation have also been studied to

determ ine the dose of digitonin necessary followingthe rapid adm inistra

tion of ouabain. R esults of these studies are as yet incom plete as various

param eters of prelim inm dosage are necessary. in sim ilar patientsthe re lationship oi

‘

the sit‘

set oi digitalis on the heart rate has hem studied

before and after vagal blochade with atropine . it is hoped that the inter

relationships of digitalis dosage to potassium , calcium , and norepinephrine

m ay be determ ined. it beam s. from the pre lim inary p ork , that potassium

increases the dose oi“

digitalis necessary to achieve a given ventricular

rate .

A n im portant project oi the Section on C ardiology is a long- term

study oi ventricular function in essentially norm al patients . C ardiac

output and system ic pressure have been m easured before and after the

infusion of large quantities oi blood with and W ithout ganglionic blockade .

T he data indicate that Sterling’s latv oi the heart applies when the regu

lstory m echanism s of the body are blocked. A nother experim ental physiologic study is concernedwith s hether the heart m uscle has "

tone"and

to determ ine Whether the pressure-volum e curve of the ventricle can be

m odified by the adm inistration oi various drugs or by changingvarious

hem odynam ic factors . The basic too l in this study has been a m ethod

for determ iningthe end diastolic fiber length with a Cushny lever andthe end diastolic volum e by infusion studies following sacrifice . A char

acteristic pressure volum e curve has been establishedDistensibility was not changedby the adm inistration of epinephrine or

anticoagulant rem ains after the adm inistration of protam ine . in 25

patients it was shown that this study provided the m ost useful and

reliable estim ate as to the adequacy of protam ine adm inistration.

A num ber of physiologic studies concerned with Whole bodyperfusion are in progress . H epatic blood flow is beingm easured in

dogs duringbypass at various rates of perfusion . H epatic blood flowdoes not change it perfusion at norm al cardiac output is m aintained

but if perfusion rate decreases the liver flow isan increasingpercentageof total flow. These experim ental studies are beingcorrelated with them easurem ent of liver function in patients before and after perfusion. in

the laboratory the heart lungm achine provides a useful tool for the studyof hem odynam ic iactdrs when the heart is com pletely exc luded from cir

culation. F or exam ple , duringbypass with the heart stopped, digitalishas been given and it has been shown to have a m arked effect on peripheralresistance which is not blocked by heuam ethonium . Sim ilarly, change s

in peripheral and m ay be m easured in a sim ilar fashion.

With the onset of there is an imm ediate fall in peripheralresistance which then gradually rises duringthe rem ainder of the bypass .

The effects ot‘

hm sia. various drugs and carbon dioxide on peripheralflow and resistance duringperfusion are under study.

Num erous previous reports have dealt W ith the technique and

application at lett heart catheterisation in the assessm ent ofpatients

with various form s of heart disease . A s of this date. 900 transbronchialleft heart catheterisations have been perform ed W ithout a death or serious

seque l. There has been increasingapplication of left heart catheteriz ao

tion in the study of patients with congenital heart disease . P atients with

congenital aortic stenosis have been ofparticular interest and theirassessm ent by this m ethod has probed a valuable adjunct in m anagem ent.

indicatora dilution curves with left heart injection are also beingcarried

out with increasingfrequency. O ther m ethods oi left heart catheterisa

tion are also beingused. P ef‘

rcutaneous puncture oi the left ventric lehas been carried out in nearly 1 00 patients. T his technique is particu

larly app licable in youngchildren and in those patients with severe m itralvalve disease in whom the catheter cannot be passed into the left ventriclebronchuscopically.

‘

in m any instances a com bined transbronchial and

percutaneous approach has been used. Selective angiocardiography bym eans of direct puncture oi the lett atrium or leit ventric le have beencarried out in a lim ited num ber oi patients . T his technique shows great

prom ise in the evaluation oi initral insuiiiciency. the localization of aortic

stenosis. and the detection oi leit- to- right shunts .

repletion» the levels of these enzym e s returned to control levels in theold anim als . old rats were as capable as younganim als in restoring

plasm a protein levels after protein depletion. T hus. there is no evi

danc e that age interferes with the ability of the liver to synthesiz e these

specific proteins . T he effect of protein depletion on the rate of synthe sis

of other enzym es is being studied.

3. P rotein depletion did not induc e the sam e type of change s in

liver enzym es of the rat as did aging.

4 . Usingtransplantation. extirpation and graftingtechniquesoit has been found that the am ount of

"

juvenile horm one"

present in the

developing cockroach is dependent on age . T he “effect of age of a tissue

on its sensitivity to horm onal influences is also under investigation. Byrepeated transplantation into successive adult hosts , c ertain tissues

(organ discsloi the fruito fly can he m aintained for longperiods of tim e

without undergoingfinal diti‘

erentiation. since the adult fly does not contain the horm one necessary for differentiation. By transplanting

"aged

"

discs hack into a larval host. ..the effects of age on suhseouent deve lop s

m eat will he studied.

5 . E quations have been derived for new order and first order

chem ical reactions in tissues which will predict the distribution of

m etaholites in the tissue surroundinga capillary.

6. T he light transm ission changes induced in m etaboliz ing sys

tem s oi lum inous bacteria hasbeen shoe s to he due to a chw ge in scatter

rather than absorption. A n adaptation of a light integrator has been de a

veloped for the C arey Model It Spectrophotom eter which perm its the

m easurem ent of the ahsorption spectra of light scatteringbio logicalm aterials with sim plicity. proficiency and accuracy,

7. The general theory ofm ortality rates has heen extended to

include estim ates oi the m amm oth rate of inherent aging, the m inim um

rate of environm entally instead aging. and the am ount of reserve func »

tienel capacity at any age . P relim inary calculations. based on m ortalityrates in

"

good"and

”poor environm ents. prefi ct a m anim um life span

in the hum an oi appromm ately its years .

8. Short em osure to high tem perature o C . ) does not appre c

oleniy alter m ortality rates inDrosophila. hut there is a perm anentattest induced sr esposnre to teniperature shore n o G .

3. T he average latency ot‘

the plantar flenor and superficial

abdom inal reflexes was greater in old than in young subjects . In

e lderly persons , the lowered excitability of these reflexes , as deter

m ined by increased latency, is believed to be due to dim inished centralexcitatory proc esses which is reflected in effector system s governingreflex activity, rather thm those re lated prim arily to sim ple voluntarym otor perform ance .

4 . Studies of the m echanic s of limb m ovem ent have em phasised

the relation between am plitude of swingand m echanical efficiency fora series of rapid back and forth m ovem ents of the arm around the shoulder

as an axis . T he greater m echanical efficiency at high displacem ents is

interpreted as the result of a greater proportion of"free swing

"at these

displacem ents . T he greater efficiency ofyoungas com pared with old

subjects is interpreted as the m ore sim ultaneous recruitm ent ofgreater

numbers of m usc le fibers in the m uscles ofyoung subjects . T his is

associated With the observed greater acce lerations and higher swingingrates in the youngthan in the old subjee ta.

5 . Methods for the estim ation of dentrans of different m olecular”

h eights have been standardised and a num ber of dasiran fractions with

a restricted band of m olecular W eights have been prepared by reprecipi

tation techniques for calibration purposes.

6 . T he tim e course at the developm ent at increased urine conccastration tollon ingwater deprivation has been followed. O sm o lar UlPratios ranged from 1 . 59 to d. 28 with an average of 3. 08. in alm ost halfoi

‘

the subjects studied. water deprivation had no system atic effect on

urine osm olality. The factors involved in this failure to respond are

being investigated.

7. The m azdm um osm olality oi the urine dim mshes sim iiicantlywith age in the rat. in the rat. dimM shed osm olality ofthe urine is

associated with an increased proteinuria.

d. in order to test the effects of socio- econom ic and educationalstatus at subjects previously studied from the infirm ary population of

the B altim ore C ity h ospitals. a sam ple of individuals (aged 30 - 96) stilllivingin the com m unity has been recruited. T hei s subjects spend a

days in the Baltim ore C ity H ospitals and are subjected to an extensive

battery oi physiolomcal and psychological tests. A ll are recruited on

Serial No ,

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finetltwta £ 0 : publicat igm ia the full HEM amamak repent , wwakky t apouta

atemfiz fimg aeleetea infommaeflwnml ac tivitiea f®t aha Office @2 figseaxch

Infiermatiemo mime aad tk@ PES Eaioxmmtimm Ufificeg amd an annual rapext ea

the acfiivitimn of Ewart inforfiatacm Cantat a AM@ag fipQC ifi l r@p@rt 3 pre e

pared « Ufa amrrativm summarie s flog she SUS t O fiQEy Q? UUEUan h@ax& suz gaz yvrheumatic fever ” arterioscifirasimg and hypef zwflafi@aa rmpwz z s raqumsaé d byafimbers mi Congrsea mm the UUmmaawfia and A labama contributiamm in heart

roamaxcb ; repertm an outstanding XQGQQI ch ascampliehmmmt s mi aelectad Ufllscfiemgis&3 ; a report an

"Iatermatioaal Ebé acal Rmmearch a

“highiightflng

fareisa contribuziama ta pramant dfiy kmawladge in gha field wfi cardiUvascular medicine s far the ga u ze W h am m M a m a Upwm tmm o w e}

finally” a repot fi é fi serfibins and avalumaimg a ywat°s traflnagghip c®mdustaé

by an m : cm M fl ce 9 5 M om s-sh Inferm gm m

Ea $ddi t£@a ta mmpmrtaa fihfi Umare Emf@rms ti@m Cafltax pagaici u

paead axfieasfivefiy in davelapmaaz Q5 m@&eris fia xeqmfitad far badgwtaty aad

apprept fiatfigm‘

mmetera o IacladaUamamg thama dacwm&mta wfi z e eh@ Ufiraca@z °@epaafing agatfimmmt £ 0 : hearings fin Chfi Kmez imute

°s fiscal 1 960 apprapri@~

tiem; eampfilfi tieu of highfifighza efi rQaQQgea pragream mafia ia Easmiamsfi a

comducge é ave gramt o auppsreQUinv&stigatfi@fle duriag fihs 1 9 58 calemfiar yaar ;

a cagpilatiem at th@ higfilighgm ia pr@gr@m davel@pm@mfi which Gegflgt fid dwrfingthe yeax; campalfisigm af haat g xe fieaxeb Uighlighes coverafig aha fireg

gnarfiar Qfi 1 9§UB required flat the Sfimage haarimgs ma fihe fiaacal 1 959

apprapri&&i@m and a ap®ai&1 status t ep®ra an hyparmemsigm submit tfid daring

enese hemrimgao

Publicafiisma

Saveral wmre iasued fiuxing aha yfiar o

Bs@rt Uragregs a 1 957, a cgapifiatisa afi items on gekec zad zasearch fiimdiaga

repartad dat ing thQ year frgm ifiv&agfig&fi1 @ms cmaducged and flappertad by &h@

EQUEUlflfltltfltfi o was p®reiculaxly umefiml as ab mwfias cf suppfiyflag fimfi®rfia n

tiom Um rasamt é evelwpmfimfim fim cardiavaacnlam EQSQQI Gh o Writeem in au§ a

ficfiemtfiy mana geshmicfi i languagg figs umdarsfisadfimg by aha ggaeral publfie

fig was QEQQ ef anteremm ta pt efiaaaiamals o A E@@ raleased wma a 1 1 0~p&g@

bibliagraphya

liszaé Uh§ fie

pref@asifim&1 j aurmala darima gas pet i©dJaauary fihr®ughJmfi e £ 9 58 0

Owner pufiiic&s£@as writtefi mad publimhad dwriag gha year W@r@ A Uriafi 83 m

twig year se a ca@p@xafiive venant e Ufi th tag AHA was tha mew versi®a

Cat é 1 @v&scular Ufiaaaae in aha U 8 0 : Faet e afi é Fi guxego- a .. r ‘ - .A

Sgeechas and Uxticlas

Spaakiag taxfig a&d maxrafifive @ut 1 i@e s wwre prepanefi §@z mmmbor

of se gackafi giV @& fim c©ajmmefiiom'wiah v&rfl@ws gw@aga fifi é A m za

8flxfi@@m pf fi fifi t Qfi fifi B me t a iawuad dmrimg ahfi ygfir o TwaRV @ 9 2

th@s@ raportad raaaaxch adwaace s wade im'

fim@x& Emmtitute

chxaa fimmoumced app®£mtmemta mmmhorahip am mha mmfifianal Adviaary fiaara

Couwsil, and awe gave b&ckgx@umd infarmafiimm cwfi cezmfimg an fimstitute

ocimaz ist who wan a maj er awaxé fiat ragaaxch aahievaamnt o fiive oi the

relegaes imV@ 1 ve é ccepwt attwa with scianxiffic ergamiaazioms b fem; bfi ifig

relm&m@d at the amau@1 ametimg cf the Fadfiratiaa wfi fimexican sagigfiiaw

fer s srimaatal Bielogy and ama by the far aha

Advaacsmfins at Sciance o in adé itioa to the relefimaa, ewa background

statemfiata ware praparcd on mearfi Institute raaaaxch invastigafififima

iflvelvfimg aarmml velummeer subj ecta, for ismuaacs ea aha intaxmetmé

press amd athare thrsugh the Clinical Ganaet Z afiwmmmsfism Ofiiiee o wwtkflwgthxwugh aha madia ef taé i® an$ aalavisien, agat e Hmfiemmmgiom Canter

axxamged imeervigwa £®E the fifireeterg fififi » om mach programs ma Wfic‘a

“Eightlim@"and of kmerica

°s

"fifirlfl

'

fiide Ewgliah flhg w5

for NEE°3 A saiseant Biraczar em

”A sk- it o fiaskat”; aad fi®r an mmx vfisit img

scifimaiac an thQ ahwwo“In, @ur fiawm

"o fismfietamce wag alga givwm in

coorfiimatimg a heart surgery gelfica@z fx@m Wim $9 an audiemae ax aha

t ae@ma AQAS caavamtawmo

ggrwaqgg

Emmafious reqmmsts fig: infozmmaiwn Exam aha gem@ral publfic g imdfiw

v 1 &u@ 1 @ a@ . m&fi1 cal amd health prwfsaafiams , mad prfivate and puklfic

mgaacies amd urgafliz atfiama‘wara recaived aad expgdifiiwusly haadlgé dwz img

the year o fimmy thasa wara effiectivaly by fimxmishing pargfimgaa

leaflats @r fi thar apprepxiafle pmblicatieaa; others ragnflt efi spacial cfi fi w

pilasiem afi infommmfiiwm or amtgz ials ecieatimts a a?

preparaiiwa of raadiug aad ragerenea liwts o p t imt xefierfinc@

tiwn‘

maiueainad by the Cemfiaro e@mtaifi img papara @m raseaxcb fiimdfiagaresulaing tram wax cwmduaead fimfi gmamfi

-aifiad has px@v&é

fimcraasingly asafiul in wi th fiaiarmgtfi@m activigfiamo fihg apo m@ °

data card files of maferewcaa ea Eastitmte a cammac2@ é publicatfi@ma emarfimfifiyappmaximg 1 m. medical anfi aciaaz ifiic jwuragls havw fiaam particulaxfiyvaluable in facilitating tasearch rapargimgo

lw

Qevetal apecfimlfiz ed prggrams'war& dasigfiad ta give t©

visizimg groupe o fimgfig ahQEQ were thfi Virginia Mfialeh Ofificera fix©gfaaga ergimimg sesaian £9 ? a group af pasfigraflnmfie cfi rdimvaacuiar pr@

=

grams for mmmbera ofi thfi S®$ial Legislat£ @m Emfi®xfiat i0m Sarvic@ 9 ah® W@xléHealth 0rg&miz atimmo the Amaricgn ‘é sseciat iau for ahaSciencag fiaur state haara assac iatien gKQups g and grewp of awaxdo wiamimgotudaat s o flhe3e pregramm ifi clmdafl lfibwratory dmgafi garatiens p lfi caures fi

saut e , and meetings wi th scientiaea “mg have iatgt eats ia cammfia mimh ahavisitors o

Thirtyo ffiur hay rfipx@m®mt@aivew firfim Sfiatg heaxm

aaaoefiafiiwfla paraicipmtaé ia fear wwz haart aemfimara ag pgrfi gfi &h@i$oxflmmtatian and txafimiag for haama asse cfl&ti@m wark o Thraugh thr®e ygaxawf tha8@ aemfimaraa xe eetvoflr has beem builg up of heart agmae igtiam

pa®p i® throwghgut aha cannafi y wha canfiimwe @x@hmmg@ wighfimé i®®k t@ @b@ fiegt k Infammfimiflfi Cemfie t vamfifiug in€@rmgti@m fi e z vieaa o

Doportmont of Health? Educatiomm and Welfare

Public Health Service

Bureau of State Service s

Serial No . 6

II . RESEARCH PROJECT dEPORT OF PROGRESS

FISCAL'

YEAK : 1 958 DATE : August 1 , 1 958

l o DIVISION 2 0 BRANCH

Special Health Services Heart Disease Control

k c DIVISION PROJECT NUMBEROperational Rosoaroh SHS=HD- 3

PROJECT TITLElo o Angoleo City

'

Civil Service Employees oardiovaooular oiooaoo

aouoy e

INVESTIGATORSDr . Bruce Pickoo

Mo o Philip Enterline

LOCATION OF PROJECT 8 0 STATUSLo o Angolosg Oalifornio Completed

PAHTICIPATING ORGANIZ ATIONSCalifornia State Healon Department fooilitieo

Loo Angoleo City Health Doparomont a

personnel and faoilitioo

University of Southern Golifornio personnel and facilities

A . modiool officer assignod by tho Control Poogrom continued to

work on this project during the past fiscal gaor o A paper on hypooa

tension was completed o This revealed that about 1 2% of the Loo Angoloo

Ciwil Service omployooo inolué od in the study had hyportonoion3 of

onion a third had heart dioeaso o subsequent to the first exomiooti om

mortality among hypertensive white moles diagnosed as having nommal

hearts was greater than among tho nommotonsivoo in the LO= 5o age

group but thi s difference did not hold ooder go or in the age gocup

55 and over o It was oloo observed that hypertensives in the age

group AO= §k on the initial examination developed coronary hearo

disease at a greater rate than normotonsivoo but that thi s held oo

a much lesser extent in the age groups unde r no and 55 and ovexo

The difference in prognosis for the hypertensive s in the age groupAO=5A as compared to other age groups is int eresting aod will ho

checked on by further folloo=up o

A paper was also prepared on the relationship of physical

exertion to the development of coronary heart disease and it woo

concluded that there was no signifi cant relationship between imp li ed

j ob exortion and subsequent mortality or the dev elopment of coronaryheart di ooaoe o Thi s oe ems to contoodiot the genooally bold boli of

that physically active persons are ao- loos riok of o volopiog ooz omorg

oioooso than more sedentary persons o

asparuusus of Health , Education» and Welfare

Public Heslfih Service

Burssu of State‘

Ssrvices

Serial us e 8

II . RESEARCH PROJECT REPORT OF PROGRESSFISCAL

'

fiEAR : 1 958 DATE : August 1 , 1 958

20 BRANCHSpecial Health Services Heart Disease Csntrsl Program

sh e DIVISION PROJECT NUMBEROperational Rssesrsh SHSc HDb lO

5 0 PROJECT TITLENutrition Study

-

sr Seventhq y'

Sanitarium

CarlJo Mssisnfslda MsD o

Miss Marjorie Csutsni

7 ° LOCATXON OF PROJEGTWashington, 9 D o 6 0

9 ° PAREICIPATING ORGANIZ ATlONS

wssuiugtou Sanitarium sud Hospitulg Esksms Puss, Md . fusiliuiss

PRINGIRAL fiESULfiSTbs plan of study was rsvissd sousidsrsbly

'

duriug the ysss o All

msls Ssvsuuu=dsy'

sdusnsist pasisuss » ages 20=6A 9 sxsludiug usurs

psyshissris pssisuss will be studied and sssssss with coutrsls sf fins

sums ugs grsup fuss Baptist and. Msshsdiss sdmissious o 1 s is ssssssss

that the total number of pstisnss interviewed will bs about 600 °

lussssisw subsumis sustains dstsilsd qusstisns on foods sstsu

during the h ussks bsfsrs hsspitslisstisu, sociological Guts sud gusssisus

rsgsrdiug the individualfls attitude toussus religios o Ssrum shulsstsssl

levels will be determined on ssmplss sf blosd drawn on the day of

sflmi ssisu sud sures sass lstsr o Additional fists will bs obtained fsus

the hsspitsl ressrds o The csllsssiug of data will bsgiu August 1 9589sud continue fos sue to two years at the fate sf 5 to 1 0 iussuvisus par

ussk e Patient s who are sslsstsd for tbs study but ass di scussgsd fuss

the hospital before being intervisusdg will be vi sited and isterviswsd

Department of Health, Education” and'

welfarePublic Health Service

Bureau of State Serviee e

Serial No o

FISGAL'

YENR: 1 958 DATE : August 1 3 1 958

2° BRANCH

Special Health Services Heart Disease Central

SECTION kc DIVISION PROJECT NUbEEROperational .aeeeareb SHSw HD= 1 2

5 ° PROJECT TITLEStudy of public healbh nursing services be cardiovascular disease

patients in Memphie=5helby'

00nnby9 Tennessee o

Mi ss Margaret Denham

LOCATION OF PROJECT 8 ° STATUSMemphia=8belby Cqmnby, Tenno

PARTICI PATING ORQANIZ ATIONSVisiting Nueeea A eeociabien facilities

Memphie=8belby Qounty Health DeparUmenb personnel, faeilitiee

Paper was presented at Southern. Amerieen Public Health Aesoeiabiba

and is being prepared for publication. in Public Health. Reporte o The

study ebowed that nurses employed by-

Eaxb eupporbed and private agenciee

perfemm many services specifically related. te the diagnosia of eardie

vascular diseases o it also shows that when frequent vi eibe for the

purpose of giving lmedieabion are not requiredg the generali z ed publie

health nurse in line with her usual activities performs eervieea

significant be the welfare ef the patiene with cardiovascular di eeaee o

Slides are being prepared on the basis of the study re sults be

give nursing administrators and educators some guide liaee for preparingmaraee to serve eardiovaeeular disease patients at heme o The slidee

will? alee g serve as a basis for developing a continuing plan fe r

staff education in cardiovascular disease control °

Public H@&lt% Saf vi@@

fiaraau a? State Sfi rmi fiea

Sevial No

1 1 0 RESEARCH PROJECT HEPOR? OF RR OGRESS

FISCAL'

IEAR: 1 958 DATE : August 1 9 1 958

l o DI31 81 0N 2 ° BRANCHSpecial Health Services Heart Disease Cent fol Progfam

3 0 SECT1 0& fie DIVISION PROJECT NUMBEROperatioaal Research SHSm HD=lA

5 0 PROJECT TITLETrends 1 3 prevalence of rheumatic heart di sease among cellege

students o

Dr e firmold Lo .fiielsen

Br o Douglass Thampson, American College Health A ssociafiion

Hr . Herbert; I o Samar

5 1 3 3 Margaret Evans

Locarzom OF PROJEQT 8 ° STATUSWashingtong D o C o

PARTICEPATING OHGANIZ A TIONS

fimericgn College Health A ssgciation a Student Health9 5 1 1 0 participaging colleges personnel and facilitiea

During the firs@ year (schoal year l9§6=57) 9 informatioa wasrepermed on 829 820 entering {feshmen by &A colleges and univerai bies o

Anfilgsis of these data revealed that there were studente havingeither definite rheumatic heart disease or hisfiory

'

of rheumatic fave?

with QQ heart disease ? for a rate a? 1 5 0 4 per 1 9000 students examingd o

A ll of the se would be considered as eandidatea for the propnyiacti@medical regfimens indi eated in rheumatic fever ; however » oniy

'

abaut

1 0 percant g gr 1 28 students are @u?ren@ly fgllowing a program. @f

anti a streptococ@&l prophylaxis o

Thug far far the second year of the study , axaminatiwns

have been repevted o An analysis 0 5 these revealed that therfi were

1 9 1 8? studenta with definite rheumatic heart disease or hi staries

of rheumatic fever ? far a rate of 1 5 0 5 per students examined c

Of fih@3e 1 55 5 or lz ol percent ? are cur rently following a program a?

pf O phylaxia o

A paper on the msg of pfiophglaxi s in th@ prevention of recurranfi

rheumatic feve r 1 3 being prepared fo r publication later in the geaf o

A sacend § aper ahowing tha compari san $5 the two yearaU re sult s 1 6

alga pl&nnad o A rapart of tha pyogt far the first two ya&rs i s

ha made at fine annual meafiing af fine ame fican College Health A sgge iamiga

im Philadglyhi& in Aprils 1 959 :

Dgpfifi zm m of Realm , M ueamw s, am Wealfam

Publi c Edw inav vi m

Bataan of n ate‘ Servings}

Serial MQ O

DAIE: August 1 9 1 958

2° M AM HSwen ]. Health Services Heart Diseaae Contm l

SEC'

M ON he DIEISIOEG PROJECT NWOperational Research

PROJECT TITLEEmma Dakata Coronary Disease Study

lW ESTIGATOfiS

Dr e williamJo Z ukelBr o wharf, Ho Ima m

Em Phi lip EnterlinQ

Mm ,Jean PekoverGrand Farm mam : Balm ta

and six contigm us m agnu m

m m » t a state wealth Department.Earth M ata Heart Associatim

Grand Forks and. Devils Lake Medical Societiea, Nanak Bamm

Analysi s Qf detailed intewiaws ebtained 033 a mamas taken in

campleted during the fiscal yaawo film y,by private physm ians was (em pmm fia mg

230 cases of «30m m di sw ge wag m pw‘

tefi

males 35 wars of age and. aver whim 1 3

were (30 1 131 d as manifest, by mygvcardial

rs shows diffem nceg of m m iaiw

e American Public Health

Dfipartment mf &@al$h 9 Educamigmg amd'

welfaw@t iie fiafil$h Safvi fie

Bum aa of 3mm Sfiw ices

Serial man1 6

FISCAL'

IEAfi z 1 958 DQTE z August 1 3 1 958

Sp®cigl Health Services Heart Diseasa Gontral

h e DfiVISION PROJECT NQMBEROpaz atignal fieseargh SHS=HD=1 8

5 ° PfiOJECT TITLEOverwaight vs o Obeaiay as ta cardiovascular' disaaaa o

pm H o 45 , Tym llef

7a LQSATION 9? PROJECT 8° STATUS

A é heville Rese&rch Fomndation

criteria foz '

new'

aasas of heart disease were prepared by'

flr o fiiels&a

and Bro Tyroller .

The Aaheville Labaratgmy standardiz ed serum. cholesterol d@tefmimati@n

exg hange of serum. $amplee with the Framingham Laboratory ;

L B OMO cards analysing Fesults of fi rst year examination maz e prepam fi o

Regulte af'

second axamin&$i@n on 0&nfion plant were aé nt to thig

f ice . Camplete followaup was done an 1 9 589 or percent 9 5 tho sa

wgmkers criginally axaminfid o Follaw~up status of the remainder? megs

1 9 new being dame by Dr . Eywollar . Thia inflammation pertains to faaaan

farnno examinatioa such aé s refuaal of examination, temminatioa mf figbgabsence an day

'

af examinatien (will be called for examinatign) , retif e c

No . NHIa El

1 . General, Med . é b Exp . m e ssy) .

2. Experimental Therapeutic s

3 . Be the sda, Md .

m s 6 ' NIB

Indiv idual I’ro j ec t ReportCalendar Year 1 958

Part A .

Studies on Vssosctivc Substances (items not covered in other pro j ec t s) .

Primcipsfi, Invesmgasom A . Sj oesdm s o NJ) NM ) .

0mm Investifi coss : L R . Geosc, L. Gille spie ,Jan , M.D . , T . P . Waalke s ,

9 m ail W m‘

bss’

g, amass sm sbes ; S. Udenfriend , Ph D . a

3 . M em o , fim w bis. 0 . R . Cra wling, (LOB) and E . Labro sse

(W ) . Eecbsissfla G . Evfiuzellemberg, D . Watts and E .

Easssb (sm ot sm ssmc)

Easies t Days :Tom i :

o iessiomsls

Osborn 1 .50

of noxepM spbsims (som ssmspbsme and fio m sbosy=é o byfisosymm delto ac id) have

been é m sssszsfl by solvsss m ssscsiom m s pspss cbsomseogm pbic techniques31m she am ine ofi pssismcs with pbcscbsomosysoms . Sim s milligram- quantitie s of

the se m gsbou sss sec presess m such ssfisc o compsm d co microgram-

qsss sities

oi the pu m p a sse s ses sfimivs arms spesfifiic s sssy me thods for measuring these

substances m um be (wise useful. Issem istcsc but insensive inve stigat ions

in mem osokogy bsvs sob been m osssfisl. This is discussed further in a pt oj ec tof the h a s, Maj or problems stag( 1 ) lack of s. spec ifi c assay procedure ; (2)emcm zfiozs of mom csmspbsiss psefiomfimscely as a comj sgsts ( sulfate probably)resumes tbss sow hydrolysis be some buns m foz fismssely she Esee fom of the

compound is m sssble m oss m ess cosdm iomso also; (3) siffi culcy of separating

the mcsbosyo scw firm s carcass phenom s ssfis

‘

is of assists . Since many other labors »

tom es w e mowworming cm was m fibos’

lologflc psoblm o our: own efforts in this

sixssafiom will tapes: ofif.

Since Gum sbylszfiom sppsss’

s so be as impom smfi psoscss in aha physiologic

im ccivsu os of mosspimspbsfims g we have been m esscsssd m studying this proc e ss

in psu smms . Obs approach m m bs so admflmfiscss lssge m ousse of catecholamine

which s ouls be Om shylstsafland mos be m ssbou sed by cabs? routes such as

monom fims «m ids ss ; chess msssmss abs Qo mssbyl messboM be fin she urine . The

efficccs of vssioss snags on chic sm cssflom m s {the O-mssbylscmg activ ity o f

the bypssces sivc fissivifissl m m be of fissszess. s limmasy studies were

with fiscpsoasxemol (Esopssn gisss ssbifisgmsuy ( 1 00- 380 mg in 1 2h) . The

O-methyl mezsboM fze was M essififis é lm abs sums m m is not esmu y measured.

In the fi rst sm dyo tbs comma swims also sppesxsd to contain O-mecbyl- Isopsel.

Subsequently, this fi sfl sg comm soc be sosfi m sd.

8. M etal Mod e Exp o Teet ep o

2° Experimental Therapeutico

3 ° Bethesda, Md o

PBS - dull

1“ xdividual Proj ec t Report

Calendar Year 1 958

Part A 0

Pro ject Title

A ction and Metabolism o f Dm gs o

Principe ]. Im eetiggtot s Louie Gillespie ”Jt o g Mona

got-foes: lovesti é atot as Matteo: Lo Tet ty, blot and A lbet t Sj oerdsma » MO D Plu m

th efeeoioflzela 0 o8

Other 6 o5

Die t ing toe pest peat we 32m m under-a

omefisye effec te in men of a ye t

expeodlimg gtoep o f chemical agents which at e claoofifi ed as monoamlne

oxidase inhibitore o (Be t etmoieo have been alm ost exclusively confined

to patiente with tweeteoofioo to that eom oottemt with biochemical

detem ioe tton of the degree ot mooom ioo oxideee (m 0 ) inhibition with

tteoe vetiooe ogento o white is deee t ited in another proj ec t description,

we might also dietegmulne any poeofiole bee eficiel e tioc te on blood 9 28 8 6 0 3 6 0

To date we M me completed or: are to the pt oceoe of evaluating tee folc

lowing em ge s iptomiez iclg (Mat ofilide fiotm LeRoc e) 9 le pheoyla b hydre z inom

propane (335 1 6; ot Lakeotoe) 9 2 o f the How ie alkalotdo a Hem line and

flormioe a Ot ttoot ine mommy9 M ot ooioo Moxie?) 9 M oh amed (Efi z er) and

We l5l¢a (Mam et Lambet t} 0

some A lttoogfla ot tlnoototic Pom otoootve ottecto were observed to some

et tefot wi th eevet el of tine aboo e e oemtiooed egemto g zle t eodily became

appet ent toot a t eptoo‘

loeible and m o toiloeo M e et ing of the M ending

blood pt e eoot e could be otteioeo Witt ooe of toome fioflettitot e g namely »

M 5 1 6 fl oneegom nyo we extended out atedies with this din g to 1 0

hospitali z ed patients 9 demooottattng in 9 a oo otainea} ot toostatic lowering

of the blood pt esem i'

e o Elect potieot wee carefully tollowed clini cally

as well as from a labo t etoty otenopoint for toxicity .) A s soon as it was

3 Serial N0 0 MRI- 22

2 months on placebo ° The results of thi s ctudy'

heve been discouz agingo

Changes in. the medical pet eoomel at well at a smallet - thans expected and

inconsistent patient popole tlom. fllmlnieeed the effoc ttvene es of tte study °

The results , howevet , to include o mombot of eeeee of good response during

placebo admini sttatloe . e e'well at telle t e to respond while on the drugo

A t this point it chee le. eleo he told that pilot investigation on bo th an

inpatient and outpetleet beele te e been cat t led out to determine any

possible aotteogloel effects eeeeltlmg temm .lt5 1 6 afimlnlctration o To

date , studies pet totmed co 3 eotpe tlemts end 3 lopatleo ts fail to show

any significant thetepectlc ettect o

a o Bfte¢t§ _effl

fl9 leetbittee em aletelet n eetetenin levels in_man~

Studies wet e pet totmed le ‘ S pettemte o Serial blood samples wet e

dtewm toting a comttol pet ted and them dot ing a period of iproniaz id

administt etioo o ‘t elgnltlceet else le . t%e eetotoein levels was

noted in all patteetc dogleg lptomle z ld admlmiottatlon , confirmingto men elmllo t t eomlte obtaleefl lo emlmels o

A . nnmbet of hypet temelt e pe tleote met e lmto oeo wi th not epineptxine

at a cooetae t t ote one dotage to detetmioe the standard ohyslologic

(pulse and bloot pt eecet el t eooooee in the untreated state . Presently ,

simile t lofuelome et c plot t ed to cotmoteoetwe individuals , The se

infusions will eetwe ao cemttol t olt ec tot eubfeqoent studies in

notmo temelve and typetteoetoe tmdtt tdualo who have been treated wi th

Mfio inhibitot e o

C o Extensive studies tot e been cat t led out in cooperation. wi th

Dt o Donald Game of the Eodoct leology Sectlom on a patient wi th

idiopathic poetotal typotemelomo Ptyelologlc studies were performed

which eoeble é '

o e to localiz e the leeloo to the tympathetic ganglion

and ctet et te t lz e to some eegt ee the biochemical deficiency , Fut toet ~

mot e ” a substeeco Foes boom ifo in this potlemt’s serum which inhibi ts

mogwe ttenemteotoe in. a ttog met t e e meecle peepet ation and perhaps maybe related to the etiology of tote patient

'e eymdt ome o Studies are

now to pt ogt oe e to attempt to tet ttet cte t octet lz e tois substance o

d o lo . colletote tlon'wltt. t t cJote . @tll of the Endocrinology Section

we have otodled euottet patient extensively lm. ee attempt to elucidate

undet t hat coodltlon ctlotothle z lde Gntut ll) will cause a marked typo

tenoive reopoo oe o The recolte me t e not entit ely conclusive , but sug»

ge sted that the degre e of typoteo otom ptodmced related directly to the

amount of totaeelom loot in. tte e t lme occut tlng dot ing bicarbonate

infusion o

Serial. HO G NHI ‘ 22

PBS- " N ifl

Indiv idual Pro j ect Report

Calendar Year 1 958

Part B

Publications

Shore , P o A Gillespie , Jay , Spector , S o and

Prockop , D o increase in blood ses

o tomin leve ls induced

by iproniaz id im. man and mashisao matwmwissenschaften

340 , 1 958 0

2° GLllespie , Lo , Tammy, Lo L Sj oeedame , A » A mew anti

hypertensive ageat , ne phefiyl° 26bydraz imopmopane o A bstrac t

and pxesemte tion at the meeting o f the Americam. fleaat

A ssociation in San firamciaoo , fleas 26 9 1 95 8 0

Gillespie , Louis ,Jae , Clinical toxiciey o f 335 1 6 admini strationo

Presente d at moooamfine Oxidaae Symposium, New York A cademy o f

Sciences , mov o 21 , 1 953 9 To be published o

A xelrod,Jo , Sho fé r , R King,Jo K 0 and 5 j o eedsma , a. o The

fate of papavez ime o J . Pha rms é z flxpfilo Themap allggz8- 1 5 9 1 958 e

Serial N 3 0 NHIu Z B

PBS -NIH

Individual Proj ec t ReportCalendar Year 1 958

Part 8

Publications

1 . Go ldberg, L° I Potomtiation of mhe cardiovascular effec ts of

the Qopamime and Tryptamino by'

monoamimo oxidase inhibitors °

N o Y; A cad o o f Sciomcoo , NOV o 21 -24 0 To be publisoedo

2° Goldberg, L, I o and Sj oordoma , A A Gaz diovascular effects of

naturally occumximg sympathetic amines bofore and after

monoamiuo oxidase iohibitlomo lo be pmbllshodo

3 ° Goldberg, L, 1 0 Use of mophonotidim (Ritalin) and monoamine

oxidase inhibitors in diffegentiafilng morepinepht ine from

dopamine by bioassayo Em pooparamiomo

Serial No o NHI- 26l o General

‘

Msd o Exp o Therap q

20 Experimental T herapeutics

3 ° Bethesda , Mdo

,PHS RIB

Individual Proj ect ReportCalendar Year 1 958

Part A 0

Catecholsmine studies o a phase of atmatas on vasosctive substances °

Jo Pnehm d Onset , Memo

Other Investigates : no hjoerdsm , h ob o , Ph o l) o

Men. Years

Totals 1 0 1 0

Professional sO thers 0 0 50

we or Findi e s

1 0 Following the demonstration o§ serotonin and catecholamines in

bananas by others in this lshoratory, a study of the e ffect of banana

ingestion on the urinary excretion of eaten and eatecholanine s was

undertaken o It use found that Shine enceetion increases in an amount

approximate ly equivalent to the was of serotonin in. the banana pulp

eaten° The free cateeholemine secretion did not increase , but the

acid-hydrolyz shle conjugates of nerepinephrine one dopamine increased

considerablya Large quantities of norepinephtine , dopamine , and DORA'

were also given orally to patients wi thout toxic signs and with resultant

excretion patterns similar to those found with the henenao This study

confirms by an indigent approach. the paesence of oesotonin and catecho le

amines in the banana and demonstrates the need to collect urine specimens

for these essays on a hananae fieee nine s

2° work continned on the neasueenent of catecholanines in'

blood, though

no progress was made in developing me thods supet ios to those new avail

able o Some comments on this week were presented at the CatecholamineSymposium. held at the national institutes o t health on Oc tober lbm ls,1 958° he furthen effort in this area is anticipateeo

ao we have continued to investigate the usefulness of eeterminingblood catecholamines at various sites wi thin the p ens cause as a

diagnostic aid in localiz ing pheochtomncytomas a the blood samples

being obtained by venous cathetenisation o Data is now available

from five cathetesi zations in four patients ( tun catheteriz ations

at Nlh‘wi th the assistance of Samuel Fox 9 Msfi o g two at the Bethesda

0 2 °

Sarial No e NHI- 27

Narmal daily excre tion has been found to t ango from. 25 to 1 30

microgramsIday o

Tryptmmioc is a good sobocracc for the enz yme monoamine oxidao é

(M afi a Whom MAO inhibitiom io pmoouccd in man wi th l-pheny1 ° 2-Pmydraz imo°

pxopé no (JR- 5 1 639 incrooooo in. uo£ oomy'

tryptomimc to times the

control level o ccors o To ooo kooofiodgo tmio incz cooo exceeds those soon

in any ochoz omoogcmomo omioc stooioo im. omo pxoocmco of MAO blo ckade o

Conooquontlyo coo moooofiomomm o i coyocomioo_oxogocioo is being utili z ed

as an index o f'

ooo imbibimooy occioo of imwootigotional dougao

‘

Wi th

this indoxa imfiocmofiiom rogomoimg coo xoflocio c yotomcy, onset o f actiong

ouzation o f offocc omo goo oooo Eoopomoo o f'

Moo iohibitoxs con be

obtaimodo

To date s ooay poolimfloooy ooodioo on one mooobolfic pathways of

txypcamioo at e oomolomo o Emcoovomooo iofimoiom ofi cxypoammno in a

potiomm woo foooo to pooooco a maoko o iooooooo io ooioozy IA A but

oisc im. 5fllooo Tmio o fiogooooo*

ofioo coo fiooiog of mo onal tryptamine

excretion in omimo of 3 pooiomoo'

o i om, oo 1 igoooc oogomtoffioomo is

pocoumotivo ovioooco ogoi ooo the foomocioo . of 5 e oyooooytmypcamino

( serotonin) fixomxooyotomimo o fifiypfiopkom. loodimg fio mom boo boon ooowm

to imcz oooo crypaomimo omoz ociom fiofioo Whom toio io done to pationto

onJB~5 1 69 quite oogo loooflo ofi oaooooioo . oro obtaimoo and tho pationco

voice oobj ootioo comoloimco oooolly'

oommoz iz od oo o”dz omk foeliog

"o

Too poowioo o ooooioo o f zoo oz oootfioo in Potiomco have boon extended

oligoolyb with goo Eamoimg of imooooooo oz omotioo io z oovocol geotto a

incoocimol comoiciooo ogooo coom opooo a ioclmoimg pomogcotio iooofie

ficiomcy coo o blood gocoooiool Roooo A ooooo woo oocomptoo to coofizm

the fimofimg of Kz ol oo oi « oosop fiooo zoo oooootiom iocmeoooo wioo

oz oocioo o It woo foomo khoo zoo cgooociom oiooo ofiaom oaoooioo s out a

ooz ikimg diooooio aloo oooooo ofiooo oxoooioo » Thooofogo g coo e ffec t

o f oxioo flow om the hoooly oxooofifiom of zoo woo oomoiod in oommol

oohj octo e omd o dofiimito oelotiomofiip moo fiomod to oxioc; as much as a

ooo e folo ioomomomo io . EoA ooopoc oooooooo after a ooooo loado

Too coypcomimo oaooy'

oill oo mood fim comj oocfifiom‘

oioh the ooz otonio

tolooomoo toot in coo olimi col owoloocioo of Moo iohibitoro o

Too mo tobolic pothomyé o f coypcomimo ggo oo omolly and IV are to befoooooz otudiooa omo moo offocm ofi oofimo flow oo . ooo oxcmetion of suchoooooo doaoomimedo Tfyptomimo ooooyo wi ll bo oomo on too oxioo cfi patientswi th a variety o f clfioicol diooooooo c

2c Loo

Foothoz ocuoioo of zoo ooo EHXA A oxcz ocioo im potiooto with inteooioo l

oioooooo ago being imiciotoo in coopooo tiom wioh the New Yook Hoopi taland Bollovoo

‘o G O E O Glioic o

fiaz t B inolodod Yoo fio

1 ° Cardiopulmonary Phys iology

2 0 M ic-dynamics

3 . Hotbox-ado, ” w e

PBS NIH

Indiv idual Pro joefi Repom

Calom‘

lm ' Yew 1 95 8

ct"mu m Tho m m mwiceafi. m m fiofi ofizim of the Pulm onary

Vascular Sm fio—mo

im i a ogtigafiorg m mJo Eafiel , M, D e

inwosfi gatom s Dom m It“ Fry e mom Donald P o Swindm '

o MO D ”

m a QM E®1 Mo Fm $330 330 9 0

fat zing finite Mono

om (Colorado? was? 39sm mu m ; mayo Kfio lomdm You 1 9 58 ) g

1 1 0)

6? Dw om pfi on

Obj eofcivom Tho s im lfim w sam m m of blood progm w o o fl ow

om ? ven om ? homo been oom flm m jly m owed from fi lo dog°s

wh amm y m m lefi oym w o PM wow m wrw om o 9& 1 t

of flmofwmoo g m m ezflw an d fm fim amoo of film oysfizw can by ,

oomm todl oomibmw sfiyo Ting offoefi of imfim fizhom o io pregam e

am igo , 153°t n m m , mifizm l im fffioiomoy and various other

intow om flom hw o boom M omm a

Maj or Fim mgm Rom lm fio 61am ham beam

(w Tho wh om {flown blood malocmyg m pm gm m am fi ne

pm sum 61m g» am em wh om m pfi h fl og o an ham

vow gfimim 1&9,t 97m in M m e Tho peak flow loads fihe

pm osunm m the M QPfi@ 1 wgfim oQilifsfif‘fig

‘fcm a m m m m

ofifsm aoo and 8m m finofi amm in tho mum m y M m ? “ in

m o pulmonary voila both m o «flim s sooflafiefl m mw kedly

domm d v m m fi ng a. very am igo m m flm oe in the pul

monary wonmw bofi o

Q2 ) Buf fing pom fiwo pm w m fls fllafifiomg fiho w omb ? m ais

fiamceo flmom oo o

K3 )”mo prodm fsfiom of mflfim i im ffi ofi moy powwow on m a y

m ow m trogm cflo fl ow in %h@ wh om vows in dog

m is; bogged m m m m ©5133 m M flfi ofl {to be wood in human

m bj oom fox? om i- qm fi tm wo ogfisimafiflom of mitral m oan

gfifim flm o M ommim m ofcundflw M afi a? fio fiho above have

boom sfia fiofl in human with om eomm fing rom lts o

Propogorflm m o of pm j oot g M M @@ of fizho m o m owoioo a

1 . Cardio pulmonary Phys io logy

2 . Cardiodynomics

3 . Bethesda , Md.

m NIH

Individual Pro j ect Report

Calendar Yoor 1 968

Principal Invest igafior e Thom B30 Em ,Jo ” , WM ) .

m inor Invoooigotom Somool Mo

“

Ea’oo N ED mo , 9 Robert P . 6m m. BM ) .

Oooooooom g 0mm Some m ob ooproofiotoefl w omen was provi ded by the

Clinic of Surgooy , NM ,

‘

You? the ir oohodo lo boo booono too busy to

allow further awe of thoir faofilifizfi’

loo .

Man Years Qoo looolor yoor 1 9 5g) ; Patient mayo ( oofim dar year 1 958 33

Total . 5 1 5

Pro j ect Descript ion

Obj ooowos a m is W fioim ooo om o om m oo W 1 11. be demo during

flofifloo ox? {zoo {coo vom m ofloo o

Maj or Fiwiogo 3 m on o oreogoooo M o boom m oo in this ow ? duo

fio {Boo foofiooo o m o fl ow filo who engagem ent mom n o? of em

m ooooofgl flooooo is the mom my of; on? a, now m omm y lab

om fhooy am {too goofifloza oz? M ioflymm floo o

”m m m m pow ido

good fiooiu fifloo m om m ofloflm of m im osa? m o} tw ining

of pom om ol M o flol‘oyool 0 6:33o fills/mm {hom o oddfitflomol intra

oordfloo oilootooolo oz oflooofiiooo 5m moo oolomfior m om

B flmomood

Serial No .“HI-5°

Studies concerning the determination of the instantaneous pmlaa

tile blood velocity.

( 1 ) Definition of nom ad]. volooi‘ay m m woriotieo at din es

ofihge imfiow omfiim g in dog am m o

gam m a of who mm m fizmooo of fino vealoom y teohnioue in

post- opsm flzivo m ilm u om of myw wdial function ( to be

a j am pmogoo‘fi: m om the m u m of surgery ) .

eluded s

l. Cardiom lnonary Phys iology

2 . Can-diodynamice

3 . Bethesda. Md.

ens NIB

Individual Proj ect Report

Pro j ect Title s The Be veloneenl end Evaluation of Bas ic Instrumenta

t ion for hpplicetion in the Field of Blood Fl ow and Preeeure

Principal. lnveetigetor s hone lo l . Frog he .

Other lnreetignioro s Seh oel h . For lllo PM ) . and Donald P . Schilder . hm .

Cooperat ing Unit. None

lien Years (Calendar year wells Pntienl loge (calender Year 1 9 58 )

2 5 G

Proj ecl heecripcion

A conoiderehly more eophieliceteo flee generelor for

loci ing end romancing ihe catheter relocily h onouring

device lo be ing coneirnclol. Thin nay-item will not only

perm it more eccmrele collection of ihe catheter tw hniqne

ho i e leo will e lloe n more c crellnl W oollen oi the

frequency coconoency oi the coefficienl oi friction in

the hlooi film of large renee lo . lhie incisor . unlike the

M oon? inoralonce a in lll— nncleretow one conei itntee one of

ihe m oheet oi the eel oi” w ow olioee lnoolm fi in the

lheoreiicel olm cione ceecrihing che celhecer- em poter

hlooo relociiy neeenrlng erelen.

An even more important m o ro cco of corolioreeculor lunct ion

then hloozl ve loc ity nozzle he che inecenleneohe blood flow

in e hloocrl reeeel . From ihe chore cloc‘

liee it in hom e

ihei iniornntion ehont relcciigy oroiilee acrece the blood

awe-ee l o ill he ohteinco one iron chic iniornetion knoc

leolge oi inelenoeneone hlooo‘

l velocity will permit the

cm ontelion oi inetnnleneohe hlocrl lice . oince the incino

leneono eori ic or hlool receei oionele r will be necessary

in three conpnlelione . cmrrent reeeorch is also directed

Serial No ,

“ I“

Individual Project Report

Honors

Publicat ions other wee oboteecize from was m oieci:33

1 ° F03 9 seam enMo H ID Edem a A ieeemder de u ce ” B od” W eedere

Cooper s m om m oe o Donald i o

’

m , mom son of the Differ

Biethods for

meet ings of the American deem deeocietiom, See M im e , 1 9 58 0

M y , dom e E.” m em o em , end m em e , A 0 an m Evaluat ion of

Modem Peeeem e deceit-ding Sm ite -ne e Ciec o Reeeeech 5 84 0 , 3.9 5“?o

Fem am end m od em s deem ” d o R a vi/90m) Modem , co Bo m Malice ,

A nd , W e id ei iom of m fi eea

Weee oi’ despim iow Fiewmetem o d o of

A pple m yeioi o M ama, 1 957 0

ew e 157m m mo gmoe Noble , e ye , and m u oe o i ,Jo en eneecrie

Device ioe flee ieeiee eee e emd centim eters cce edeei ioo oi A oei ic e

Biood Ve iociiyo fiiec o deeeee cfin 5 375 0 19 57 9

Serial No e NM =M1 : General Nad a Exp o Therap o

2 0 Clinical Endocrino logy

3 0 Bethe sda , Md o

PBS NIH

Individual Pro j ect ReportCal endar Yeas: 1 958

Effect of Chronic Hypercalcfiw fia on Renal Concentrating Mechani sm

and Sodium Conservation ,

e mote Em oegigotoz os Ban ter, 3. co . Mom m o 8mm G o

Professional

Other

1 ° To shady the effect): of figm emcalcmfl o o pam tfinyz oid hormone » and

vi tamin D o n the ability of the M oney to concemtgate g

2° To determioo tho effecfi om Coho comconm atmg defect of restoring

fl im sy calcium to m m }. m bm ozecalcinnz ic pomiermtcs o

3 » To dotom ioo whoa-hog time, oom omtcm mmg defoct is associated with

inability to com m, oodlim o

It: we been rep uted! that: patients m m W orcalcemia aiid hym n-calciu m

may lose tho ability to comceofirofie . oximo oven im. t eopoo8 e to pitresoin o

The peooemt m unchies were imoflcigato é i to dotom ioo what factors are essential

fox we development of 1};m defect ,

Eight: patiomts aim mfl

pez colciabfi o m m m an go In all: urinary

com m mm cmg mommy was impoflcoo ao jmdlgoo fl ow Echo maxim urinarycommun es? obm im ble m esa fiehydlm fizioo flow: boom m ad pict cssina or from

dotem im n om of often“ the mofifi’

mod of Z ak, 5m m and Smith

Whom oflwfiome dim team ocemw «mm » was given by mouth to

rectum M m calcium m nominal9 m o re woo oigmfl.ficam improvement in

concomtm mog abfilfimy o fiwow iwmm ooio fl and Powem z athyroidism appem ed

capable of m ow ing a m oo sw am comcootfiacimg dlofiecc for the degree of

hwem alcm z ia ammo em s fiovm d in oazcoidooio a oooonciafi Moonwalk :zlaxzs'JLa9

etc .» A ll subj ects with mo fiofoog comm cow ow c oodmm on a low sodium

Sgrial NO 0 Nfliw&7l o Gen o Med o gap e Thgt ap o

2 ° C limical Endoafi lmo logy3 ° Be the sdafi Mafiylamd

PBS w e NIH

Indiv idual Pfimj ec t Re port

Rafioatiam of the Edomfi tous Sfifiao o

Gabe? Enve otfigs toaz g E 0 @ 0 Bfiglioz i g E 0 9 Jo R a Gillofif o o £ 5 9

£ 0 B o m u s s, MOD C o 8 0 Baleo t, 63 0 Kat hy ?

G o Smikh sA OG O ? O Coopef g mod fi e Headgrsoa o

Patigmt Degas fiflfi

To fial

Pt ofiomoional

To ovaluotg th® r@ 1 @ of tho adf oaal aofi tox ia th@ godium

To iavo omigato afie o timuli to oldo et@3 @m@

20 To evaluaao the ro ia of alé oatef ome in pathological

3 ° To o luc iflafio emg mgehanism, ofi ao tiom of aldo s t@r@m@ ow

mho kidmoy o

&c To gvalwamg $h® go tiom oi aggfi ts ai th a po t é at iml

on aha o i alé ostoxooa o

1 0 ?hé offiec fi o i po tagaium Em e lavaz ing olé oatarome

and oi po fiaaaflum fieplgtion in low&mimg aldoatof one o@@fi @ tiofl was

inve stigato é in aogmfi l aubj @@ to o it woo shown that aldo 5 a@ron@

B@@f @tion coold ho olevote é upom loadimg oafi decz oaagd with po tass ium

deple tion withowfi ovidomco oi oomoomitamt ree iprooal ohfimgoa in

imtz ovaaoulax volwme o

Sarial No o I‘ll-11 4 48

The se technique s wera used to m eaun the effec t of e strogams g

of fasting, of Nilevar ( N e alphao e thyl 1 7-hydrozcy morw droatenone)and of surgical trauma cm tha degree of plasma binding of hydro

cortisone o Preliminary m aulta indicate ( 1 ) that e sta’ogen tam-Badly

inn -asse s plasma hydrocortismme almanac m am a

-M y through effec t of

incl-sw ing hem d [swam hydrocortisome m ad thaw preem bly the

m ount of spec ifi c hydxm omtfieom -M né wg prote in, (2) m en us may

produce a amn m fincm aw in specific gm z oid bimdimga ( 3) Eilevar

probably M e s am have mum m 3 5mm m em u-mid binding, (a) no t

enough re sults w e m an dala mx w zgmm tram Q8 yeah The

apegifi g binding ms hydgoaom im m in fiem l plasma was gonad to

be w ry small» a fi nding qm m in m ung with am very low level

@ 5 total hydrocgrtm m e fiom d fin gate}. plasm a

Wang 1 3 0 mm te©hmim , prelimimazy sawdm g 1m m been done to

avaluam m am e lem -mace M hydxm ofl flam by compafi ag the clam -w ee

M ultxafi ltem b fie gm f om with that ofi im am 0 Th@ m ann a sugge st

that sw arm tim e as m m Mam fim e hyd mggm ism e is fi ltered as

is em m m d in &h@ mrm o

A M Labellefi am m ida ©M gimabm firm am Eadw xfimology StudySectigm and {93 mm ? «35 amem labelmd by {fla-izbagh

38 technique

and purifi ed bet a have M esa axm fimed flaw dggm e binding to alw

humane mm firt m m? m plasm a Rw ullm imm smm that plasma

bimdimg afi aidw m rm ag pfi’

ogm m m m a and m dm stemediom is am ounted

far by the atmghm m: ofi thaw stam ida m mm m M ama, whereas , a

spas ima sw tfi aafi mfi d birmdlimg pgm mm (pm bably m aagmm m )am azingly M inds hydm gm ' tm om o m m mm a, cw tm osmm m m d comman d

m m them atafom s p appm fi abfie M mé m g m albflm m apm a m only afite z

the expcacifim 19 1m m a m amasm is m am fim aflo

m steady fla m ing w aeuzfiaél m alw fifiy aha) 1m m ofi fas ting,

team , estm ggm v m gmm ’

sfiem icgm ma apeefifi e glass-

m bindingo

Stwdm s M lmde degm m fima’

aim ©z§ hyé gm em im me pawl aim flam ingM m chm ga M ahaw d by tx;t agemm o

Studim with dfi fiem mt m ermaids a m be ing ©©fifitifim <ad in firm

atmmpt m Q1 fifi©fid®fi® aha m im icm m p Qf stem m atm m xa fig

gpcac ifim Emirfidimg ©fi m em m o

ym t B m mmfi

So t ie l No ° NH1 ° 53

1 . Gene ral Med , Exp . Therap .

2. Clinical Endocrinology3 . Be the sda, Md.

PBS NIH

Individual Proj ec t Report:Calendar Year 1 958

in mammal efid Gdefhoefic Subj ecto

sem en, m we % .D .LBan ter , F . o n o ,

G. Kareem

Poedeme Doze1

e mechomiom fior ahe eubnormol feee eater clearance of

is , To deeemmime eke role o f peoximal tubular sodium

imeefile hotmcme do produe iog this defec t . To eluw

emiems involved by peodoe img o compexeble syndrome in normal

leem .

Goin, oeel and leggeveoeoe womer _loedo : The diure tic

omse of the normal cobj ecc has been shown co decrease progre ssiv elysalt deple tiom lo incomeifiied. Am fimpeieed diuresis of free water

arable'wiah that shown by a pood leeiom of decompemsated c irrho tic s

aeec in the well hydrated moemal sobj eem efeex intensive salt deple t ion ,

The infusion. of mixed isotonic

a and fructose) has been shown

:mcxeeoe free woke? cleaxemce in bomb oderhotics and salt deple ted

gals above that seen with we equal load oi the bome se alone ° Both

ups show'

e comparable degeee of "coz xeotiom" o f the impaired diuret ic

somee to infieavemooo hexose when the memo itol mixture is need .

do free wager is fiotmed. whem ioocomic is infused into

aorme l subj ecfi codex she se comdifiiome , The decompeneated c irrhot ic

the ed it depleted normal do oboW'

pooitlve free wager clearance undo :

ye comdltioo e o

Serial No . N51 ° 53

Ban ter, Et c An Explanation Experimental

Reeemclom in Clt thoslo, Abstrac t ,

New Jersey” m y, 1 958, p . 58°

Serial No ,NHI==56

1 . General Med . 61 Exp . Therap .

2. Clinical Endocrinology3. Be thesda, Md ,


On the Netut e at Poteeeimn Losing Re nal Disease

toincigel Investigetot z Be t tte t o Ft edet le o. 9 M D . 9 end G1 1 1 ,John R M.D .

c e t Inne etigntot Delen9 Q. S. , willet g t w Illldldlettm 9 B Turner ,

Bethent 9 D . 9 end Kinny» c.

Patient haze : 250

Hypokelen in eeooc inted with nt inety potaeeinn lose appears in a numberat eyndt cmes , The pte eent etndiee were lnet itnted to determine what feature s

meee eyedt ome s have in common, end in net tienlet the tale of eldo stez one in

the tone ]. poteeeflnn lo ss . A lthough toe the pnm osee of the pt esent study

patients with peleney eldoetetoniem have been enelndedo the studie s point out

the difficulty at eetehliehing eleet ly the difi et ence between primary and

seeondety eldoetetonien ,

"two m oons of petiente have been studied : thoee with assoc iated

alkaloeie (2) a end those with eeeoeieted t ene l tubulet acidosis

Stndiee have included balance etndie e with epeeiel t etet ence to sodium ,

poteeeinn and hydt ogen lone end t onel elennence nee ennnnente .

In both gt onpe ot pe tiente , inehility to coneetve sodium and

leve ted ut inet y eldoetetone hove been denonette tefd . It was clean

nonetheleoe thet enee eelve pote eeim loee oould ocean when sodium was no t

being loot in the m i ne " (Bonne t eelya eenete eelt loss was obeeeved to

gothen with the nbllity to nnintein noenel pote eeim balance . When the

m tteeellnlet end intenve eenlet filnid volum e s not e expended in these

Se e iel He . Nhlm fi?1 . G eneral Med. 5: Exp . The rap .

2. Clinical Endocrinology3. Be thesda, Ed.

PBS m s

Individual. Proj ec t ReportCsleedse' Yesx 1 958

Ssseoieosie

Gill,John R ne e see Ban ter, Feedet ic G EM ) .

e. Melee she Seefifi

m ese em diee have heeh dome in conjunction with Dr.

firem en hell (m m , mm) .

Peeieh ee selfish eeeeeiee ees fieeqeeeely have hypeeeslemfi s even on e

hm m eme . It he s hem eeggeseee, hue m e peeved ghee ( 1 ) hyperaeg is is seem eeey he M em eeee eelefim ehsoepeioe , (2) ehe inesee sed

meme he s eesh le efi hgeeeseh sihevmy he weenie h , (3) s teroids wish

shyeee ee seeiv iey will eeeeee ee ehe hypeeeelemeis , sand (4 ) this sen se

he sees’

e ide opeeeees by M eshing seeeieivfley (to v ieemm D .

Meeeheu e hai w e e emulates have eeee eees’iee owe he shese patients with

sleesfis o heeem zleeeiohs m em e eefleie e helm ee , phosphorus balance ,

whee heleheeh

smsl peeioslie eeeem iheefieh s efi seem , sele nium , phe sphom s ,

Mem es h.

‘Ehe ee sfiee of m em e M eleee e m emes of sealed saleme

“

vim h, added eeleim pile s peeeh fieeese see ell eh‘

xee teesm enes . A

e w hee he" psefleeme hee e heem sew efyee ho eeeem flhe the ir suitabilityehie eeeeeee . heeem fieeeiem have Meleeee Tm efi phosphom s , se spoxase

he seems ease lemme eele zhm she phe sphom s go amphoj el on s low phospheee

w e eeeeeeee es esileflm aM eme .

Resales oh ehe beheme e eeeeie s eee still in peoeees ofi analysis .

M isses“

? eats M lesee ghee a) peeieh hs wish semeoidosls diffee ms ekedly

eeeee sfi eslemm sheen-

game , (2) ehee ehe se peeiee es do show hypez

N ew wieh new fieesl eelehm she? may eehfihzlh normal blood levels of

his 9 , (3) Eh peeiemms m m s lee fieeel eele iem sh e hypereelcm s is ,miasme may efifieeefively diminish ehe hypem eleih rie o

Serinl No . NHL -581 0 General Med. Ga Exp . Theeep .

2. Clinical Endoc rinology3 0 Bathafidag ”do

PHS «m NIH

Indivienel Proj ee t Reportcalender Yeefl: 1 958

Pos t A .

study of Movement; ofi Peoeefins one Lfiphee Through A e teeiel Wells

and Sh iloh Tissues

PM? Dnneen, L. h Jim , n o .

Othes Enveehfige toe ss hneh , h e , one Lyneh , A . weehnieew

An'

nneeeoeneeing oh the peoe eeees involved in ehe movement of

peo eeins end M om s finite one one off eneeehnl wells one sinile e tissue s .

The week. has been eeee zlen one hy eenoies in eshhieo end dogs . Since

no me thod enistceo foe dem ise-

mining the teenefee w he e of snhstzsnce s into

end out of them es nhen eheir: hennsfiee w hen one slow, one ee ely studie s

neee devomee to working one eneh neehooe . In {these een’

iy shudies the

mov ement of labeled nlhnnin ehzfongh enhhie hom e she tissues which new.

in some easy nonphologieemn oinihes to not es ne e steadied.

Follloning this the nonenene of inheleel ehhnnfin into and out of

the h om e onf the dog ne e seem ed}. The hom es? siz e oh ehie artery meee

n more e eeefi en enelyehs poeeihle .

c{the h om e were div ided into a. nun »

her of enee e one eeeh neon tun e split. finho wh en, sneeze , and outer

lsye z s . The heme ohhe ineal sem en; fiche coneeoe ghee oeoee ins move from

blood in {the eoeeie lenen eeeeee ehe inhineh enooeheunn into the inner

leyezc of the som e . n seehhfing geeeiene of infl ow notes for s lbmnin was

found. In ehe peoninel eom e neee . flee origin won ehe heenh, albuminmove s fines ehe nom ie now. very nepioiyo

k‘Ehhs infi en

‘

enee“

dee eenoe e

well see higher: ehen they one in the M otel hom es h ello

Serial 3130 0 2 153551 4 9

GEROI‘H’OLGGY BRA fiGH

September 1 , 1 958 a December 1 958

1 ° an Title s Studie s on the m ahanimn of oxidative phosmorylationo

no W eipal Inve stigators no Ran Sam di time )

wakinical M smwnee m 1 s wyasr

ca Pragm as Baring $3 9 3, Fm m amas 133m is a new pm jefct initiated51 1 fiQfiCM E” i t ' s 0

6-0 m mm cf alivm w a Ram wm s We mamama-

mewm m that cad=

M um im at, M agi? mu Em ails m um m mme phosphorylationm a m m m d by dithiols but not by

m m far M ati ng in mu chm drm the effeet

m aefimm {that am am m utly closely related

w pam phwylafi m g e ggs9 mm activity umm' various condi=

MW and phagpfimw=fifl° ©310m o m also pregam e to study theefi m fi of flaw ia emm m mmm z fm gm nta which have retained

ma r capacity to cam dam omdatim phosW ryh tm o

Pm pam fl by Do Rae Sau di

6& q 279 1 953

Serial No , NHL-60

c o Progress During Past mama apparatus for this studyw e constructed

m adm plua the expera

Man n a duration on 9 94 11

parim mal comm ]. of we embjeactw flam ing a particular wheatin mm M tbs source of film am é ming atim lus o

wen completed, m m mm m m pilot subj ects twang}; on aha

m m m m adam in m ay to mbtain ramming Mm estimates md

30 callabam fcwsa Baltim m

lac Publications m M amas

DoGamma? 309

2°

30

14°

b e

0 9

do

a 2 6

Serial N0 9 NHL-61

‘me subj ect is asked to m pee"£ as many words as he can

remeubero Scoringd e in terms of eom Mutation of the

M e! of words recal le d cem etly'

o Analysis is in term

of the interactions and prim e? effects of the verieblee

of age length and mmm‘

euey Since these em date willbe used in the “

beet ef meme? hype'eheeie (see Verbal Pen

formence study ) eeeh embj em‘

is m eted individually and

repetitions ere m em e em mw eie tam e Appropriate cone »

trole re possible effeeefof «meter of preeenteeien of the

word lists are Meeepem eed in mm etndyo

Principal Investigam e s ee Jo Sud (Uh Mm )Be D Davidefi

‘

((m em

Jeeee wife, W e aim wptember)

Fram es

com m). wbjeeee heve beenme date

ien of: the hypotheeie o

three ege m es have been

ingo

Directie

1 ° way is be ingm emno Amerthis paper isweek in this are a will be

instituted

1 1 1 ° Gem comm a

Patient Deye s we eppliee fele

Collaboratex'e a m um Gieggfim eieele and we ween-ere AMskeet-ion Hemfitel 8mEwe Hm m g M anda

Publicatiene end m em e 3 mm

h em e by whim D o Davidoff

eeeeeee 30 1 958

56 231 23. $30 0 8 Mil- 65

Please“ ; mm

Jam m y 1 9 1 95@ a Ewaw w 31 9 1 953

Title 3 A ge chm g m in m m m am wfim bm mm w o

Principal Inm atm m s s .m flm Q0 Bamm m

Other m um mm s ES} we ShockJo A c fi lm , ar e

3&m ww ww

Fm m aa During the: M t M lm WW W

Ob acmwaas ma mm a. @fi Maia prom is to emsnag ®f m m m 3» ehm gw w m cfiafi d with sm acsm in

tiswa est m m m Spacin g pm blem M anam a

during this mrwd ham wm s

(1 ) Oxidatiw pam phw mmm in m m 1 r

(a) m M fm fi wf ma am m 3 ability M m m m m m mm m y :

whatm fie has M esa m am a in mitm hfim m i mh tad from

wear by we m um m”? Sahm ifi wam} fiw bm mo

(2) % m www w? W am Md m m m m MW ? (81 ) m rflfiefad

m amm ly (h ) maxa m am m @ «m mfor 28 days or (6 )menthe stock (Riga of our labom m far m g days fawningmg m m of m m M MM M O Farm emm ie aetifi tias

well as Rm » 1m am pm'm im mitemwm wf liver aw kidm ye

of W m m m m m dmfsm fim d by m wpfim 9mm pm cedw

m s o

(3) h e m m us fl aw s com m ute a scribed atom s (2) ca n also

(lemm a in tam tiemzm QE 6 m ath ©la rats fed a protein

fw a diet for aim ? 0 29159 7 or 1 1; days o

Serial No e. NHL -65

in 13m m cbm m d in m m fiepleted

in the concentration of

occur inm m magma

am wss a? anim algmwim m mm 629 133m men not onlym m m m }. m lwéymm

’m l ww fiimg am ?! im am as m om

mm m m g fwfla m ifé mg mats, mg ; m ai m m am am

of mg m am m l m agma wait 3m? @m @fimm o m , m man

l M m m mwm mw W W W M adam m =

i'

z‘m fi m of mam a’s?» figmxw©ha Ffiim gm flm ms will m m

m am a {rim Mm m tag m tg ©i‘

M em m agas a

2) A final m esvm fifim gm agw iw mam a 5m m um W h amby we deapleafie é m yl mi wmambdo

s

3} SM Q M? ©Mmgeas in {gm wawznifam mm a @5 gam e in m m

{a} m W amm m am ay 4333{mam mal 03 hwy m ight magm a

fin wm amm m ime

5) W @ amiviw of Mam a ($1533?n m gem m tion (liver ) andW fl mpfi (W ay) af arga m in m im ls as M em m amao

a M afi a 80 H 23° 15 ° Shm kg and Bo F0 Chow'

s age dfi fem m e

m mum of m m and liven Jo Gam a y

Barrow sGo Ho s M0Jo images,» and BL, H o Shocks Sé mscem a and

the m tabolim of m io'm M am a at“

m ts o Jo wront o a g} ;35145 355 9 1 9580

Serial No 0 NHL-68

m inimum rate atand their m mto the m ange w

is m yfived excfim fivefiyM m pm m itm M istin g

The theory Mm mmmg m ewflm momwthe ammmt of re sgerm m agm aW N W m may awmafiamww e m flafifim flfifip magm a neg fi fimfierwpfiQR and

Gampem gnaw (as) Mags

ggmmm wm for R0 m ag ma can esti

Effg cam aam m am for m ay em inm mfig appromm fiy Law/mm am far gem mw wamem as

mmM mamfiy 0°WW’

WM O m wm m fim m gm m e misfit

m 6mm m ex Em aam @o‘

W/yw whim wwflfim ;mi: é ima; am am am m fim am m gm cmfififlafim M somewhatHem M £40mam a

m m m M m m Ram m fin: M ®E w as a thecm m gm m g M ggammy m a fifi fimm m acamw tgm

wm mewwg we mm y m ak eway ,

oam m m : Emisfiym m may fiwm afi

m m mmg am fiM figz Nam

P mmw fi by W M QILo Sfim mgrNam mf

‘

fim a» 1 958

Serial No o Mil — 69

eetlefectoriiy the future com es of phyelologica! indices to an

mammalprimer eom e 50 year old eebjecte am80 year owemmecte may have eqelm leot fooctloe even though

they are eefieeted from eroepe meow m ean m im e are markedlydefereeeo Dem-aegthe peeryew 9 we have been fortunate to re

creiae grow ofmeme trefieed profieeeaoeelm en who have w eed

fie eerve m e em fie mem ewere ee eebfleete for etudiee carriee

oerto $331.W ormowe

“meee eetofieefie five to the communityoeeefiee orfieetfieeefieee em ere w e eetwefly workingand retired,

Seefieeee ere efiaoeee by sewe r emegeere refiner them referred bypeyefiefieee or efinoeee by fiefie orgeefleefiaoe o mum my, the emb

fieefi fie gfleee e moreege ehwefieefi em fieeuoe w e the reeelte

orthee ewe may teem emeh m y be orefiiefieefi imereefiare re

poreeeaoMe oeyefleaee o“E‘

eeefleeg ew e fie primm afly orfieetee

{Loew e the eerfifioeeeeefiers m flm ooery em reeeaeyeteme , mbeee eerrfiee om era3A eeefieete firom e mworm emcee ageerem firom 23 to $

93yeere o

M eeeaoo of CerreeeReeeerefir: We em M em e to wemmy effeefie fl eoefioe eeomem fie em eeeeetfioeeaAfifereeeee befiweeetee eoeefikefim eme ewe flee grow referredm hem e

A m eeem omeaeeriee ofeeefie were em peeeie oe bee? eompo~

em ote em the eeeromeeeeefler eyefiem em be beam men yeero

A rm fioer eeee befi erfiee oi eeeee em be comm eted beiore

me preeeereerfiee fie repeeeeeo Thae em prowee repeer

eeeee every five yeere oe the ewerege o

were m eow mmthe eoeefieereeifie me newofthe member of

m fietoaee fieeowee'

ie mme m ember oi eeegeefie M fiemetedg

oafieeoreeoreg W 0 M0 Peeerg eeeefiefiC om efieeer

Sefieefiflefie" were » Port Reeemfle » Mm meed

Bebfifieefifiom m e A e er®

$2e N W ,

"

0 Som e myefioflogfieefieeeeeee of egfiego

T o

9

°C fieem ee fi (m e m e) M Newe fior Lefier W e ,

NewYork'sewe 50mgwere Lf‘

m ’

o

Legae o W N o, e reeeg pea M afi a

Preeered by m om H o N orrie

N ovember 39

Serial No o NHL-70

(0343 ) of the oubj oot o with high C:us‘psubjects are alert, and

motivato do Under conditions of vary high (2459subj eota will

diaph y symptom of he ightened affect or emotiono On tho

otm r ham]9 with low Lovell! of cm,subj ects would tend to be

lethargic ” slow roommdimgs and would ohow higher sensorythresholdo o We bo liovo that ogod subj ooto as a. y oup willtend to roll in thio m oogowm low W e. The Iow a love ]. ofCMS on thio commuum would be oboom d in deep sloop o

he mo m u vny loool of mm was o o n be oboom d and to=

oorwd in intact Em a oubj oom by m oording (oloctrioally )tho om litudo of m ou th (otm toh ) m floxos o Pm oontlyam iloblo m m opm’

oiologiool ovidomeo otoongly supports use

Of this M tphwo

so cos ond ow loom of aotivmy of the outom mio nervous

systom om rolotodo Tho lotto? is roh tod to sow aspect of

tho olootm ool phooom oo of tho okm o

Ploo of sm dioo.f. study will ho volid mioool and will inm ati=

goto tie m mfi ou of tho omplitudo of tho otm tch reflex and

tho golm nio phooom m of tho skin to oomditiomo of sh op s.room, and activity oo

gfoo om mplo s pom ioim fi on in a 3 1mm

and choice m ootion woke Wo will ottompm to ohow that those

m osm o w ry concomim fily with gooso Momma in bohovior

(i co n , amm o ffm o oloop to root, to activity ) in separate

groom of aged and youthful oubj oom o mm room agod and non

ogod groups will ho oom od o we om oo that oim ifi oont p oopdifforooooo in m oo m om m a will oppoorg with the aged groupshowing am com m on. loom? lovoi of OMS O

It is worth momfiziooiog in Maia oomoxt that. tho Comer for tho

Study of Aging m M Univozfloiwm oomoly m mrfiod that. anoldorly group of oobj ooto amm o doooooood m om oopli

‘oudoa of

galvanic akin m oponooog oo we ll as o doorooood now r of69oponm m om ‘“

golm iofl kio¢m oponoo fluotuotiom 9whom oom=

pw awith a young woup o mom findings are related to what

the im otigatoro mom o ooom ooo in oemoso arouoolg ood aro

found to oorm loto with doom aood ability of the oldorly groupin m m }. m oon o Wo om ot fizo oom otoom to and w in gs on

Mo se fi ndingo o

2° A oooond study will ho oom om d with the relation of son

son and motor dofi oit in aging to Com m ]. Motive Stato o Via

M 1 9 audiooryg and poin=throshold determinations will be made

concurrently with m ordingo of stroboh e rofl ox amplitude awl

the galvanicw kin phooom na o Sensory am motor=roaponoe effi=

oienoy over a two=hour poriod of continuous vigil a ll be

otudiod through tho use of a Hacksaw-uh “clocking Data will be

anolyood in hom e of poooiblo rolatiom betwe en oenoory and

motor function and loval of CPR ,

Serial No o s NHL -71

Jenoery lg December 31 9 1 958

Title s W eid fonction end ege o Ihym xine de gradation otudyo

Principal Inveetigetore z Retort Io

Other Iooe etigetor s m m J o Yienmt

technical A eeietence s So E a chowder1 2

Prom os Bering Poet Twelve lionthe 3 Additional enhjecte hem been

ct to the rate of dieepge erence of trewr dooeo

of intretem eely ede ioieteeed 1 1 31 towed thyroxinebloodo A tote l of 70 co n eged ld=9© yeere here been teetedo

Them ie e highly eignificen‘

t reduction in. the rate of dieeppeer=

once of thym nine M e the bla de

Sh iler eweeiu nte tee m concocted in e eeciee of old and youngrote under e m o iety of em ciee ntel con itione o These date, are

mm M lm ao

do Moe ction of Cem e nt heeeerch s The dete will he enelyeed om pro

weed for pehlicetioe o

Patient Dope 3 item

Collaborators s Beltinoee City Hoepitele peoeide epace end all utilitie s 0

Publications and Aoeroe g fiche

m oored by t o we Shock

Serial Ho o t NHIJIB

Proj ect. Remit

January 1,

a Dewmer 31 9 1 958

lo M Title s Studies on m lationm ipfi batm an dam lapm nt and agings

IL, Human ]. control of the accasaory an glands in the

cockroach f, Parigl'am m am ricamao

be Princim l Immutimtor s Dicatrich Ewem wfln (l% )

Other Investigam 8 I mbalh Spm gm

(m o Holgaka (Bel uga National Science

Foundation Fgu owahip )

c o Pragm ss During Pam, l Homthm Thifi projact E‘

QE begun about

Septamwr 1 9580

Dirm M m of Gm m nt Ram w cm This preblam has baen attacked bywe use of wm plammum g axtirm fiian am grafting m ohaiquee o

The ow m fiory am iss-my at the sm om‘m v female am : glands is

camtrollad by the jum mna mom m a produced by the 00 1m m aw e

flam e, Thum pby {twin ning the m0n=fm tiaming gland of a tsunam i

adult Mama mm the bow Qavimy of a aeries of w hen Qf know

was om em m essy am 51t Mom m a law }, in the host body o

mm study ? alw adg Quite far advanced, indicatesa that the hora

m m 1 9m }. diffem in m im h of difi wm nt agao

A mthm related wabiam m ay inflammation is to dwtermim at

what sa ga during the m acamkwym ic Mm lepmamfiz of the insect

m e: am : glané e are com mute m respond to num m l inflm m eao

mm is 109 1 3133 audited by tram plm tmg glands of difm m nt age

into 23q t and adult. beamg1 0 9 0 9 mm animals of different

humeral envm m mts o By the removal of different. glands of in

warm }. secretion am also able to gamer the humeral environe

m at-9and to use w eb altam d anim m as how. for from sex gland

2° Pé tient Dam s Mom

30 Collaborators a Home

ho Publisatioaa and Award“ Nam

Pm mm d by Diewich Bo® n$tain

Octobar 309 1 958

Serial Nb o z NHI~80

i egt Damn

January ll.s 1 958 e 126 e 31 9 1 958

1 0 m mum Studies on m k fi am hipe mamas», daw loxam nt and agingV 0 Ram and m atch atrophy in ma cockm cm Perk :

b e Principal Investigatom Maw-mm Bodeam m m (gm

fi chnical Ansiatam a 8 mm ; Gabbay (55 )swam He lp

Progress During Pam mu m m am as h is pm jw t wa s begun inJamGawain m m }, muscles of adult cocfim achaa can be m nderw

ed fw tionlsaes by ms saw m ill. of tha diam ]. part of the Iago

In these w ees it hm beam obm m d that the warm s innervatingtam e m ach : W NW aatm hic am m go finals affact. might be dua

to a reduction in the d m f 92“

um M ividm l m rw fibers or

a h as of fi wm o (gam ing-y m mm mmm m whether tbs

inactive m aclm madame daggm m tim fl am es ands if 809what

sort, at changeao fi mm 1 3 m m aviM amfi) of m ole shame cos h

do Dim etion of Garm ut M m am m A lama eeriee of em fi m ma have

beam am; up to inw m figaw finis o mm is a M ag=m nge proj act

wince it. tam e m mm mmm 11;m afi acem m en u We mmm to

m cp ifi ce Mam w in ks m; dfi mm m M m intervals and studythese chew s histah giwm and hmm ahamim uy is; Gu an o

20 Patient Dam s Mm e

3° Collaborm ors

W m te in,Dietrich (in calh bom tficmwith V Dathe ir ) : Hunger in

th@ bMWflyo k it £ 0 rh tbz l z' 1 5 3 1 29° lh0 9 1 9580

macaw m arker of the fiafiiom l Aw dsw of Sciem eao

Pram d by Dietrich Bodam tain

October 30 9 l%8

Serial No o NHL-61

action of Current Rosem The hem oglobin m olecule willbe omotfoct of each dfioseotiom

oogmm etrgwififihe do it is hoped to this mayobtain an accurate pmcom oog the hom e within hem ogfiobfio o

m om ma-m of hom ogfiogfig mm gfifiog mum followed by rotatoryrgfioo tecfimfiqgggo

mam »otgdfiog mmm om rfigo

‘

fiom om ogmfiaggg poxm om e ; chic: ophylfis

F a Ggfim gg H ofiafiomg dfiggom fiog of cgfiochrom e mao

QM »

Prega mo by Gamm a? Lo Emm om

m fiogor 31 9 N M

Serial N0 0 NHL -83

Mitochomdrtanstudies For the em em eny interestingm eta.mo?Tic

‘

M-‘

mgsg see t o Barrow “report 0m preparative

technique is m tmafily identicaltomm m ed byWeinbach and

GammaM rsom lcommamficaiom from m W eimbach»so thatdfifi erem eg m rem flw «swarm m emmam d on thaw1m m ,

RNA M m mm m flww as; fm c fiaw from the am 1 0 ram"

is vé‘

w'

cm pfiefim mm u°m w am fiaflfiy the sam e as thugs

M en-mad prwfim fiy m umm aam fiym 05 these data is 1mm

com pfiem an 33m m? mmm m ag m y m em aim atatem em alt

mm Mam,

N 0 Em ma efi m gflaw ofm mfirm fifiammmmtecmmfiques havenew gm, q ; gam e mm m pm a

do W emmm of (m um w mh d m flmchomdrm

Mm aw fiem fig 3mm wwwwmmm emmem e aflm afiy gmdfied

M fififiQM OQ GE fiQfifiM® hm aagfim flmfl fiwfig $0 ratugguego

M fr-if m wm mgm o

B, a fim m mmm fim GEgfim m g 3mm m m etabolic

amfiflm andm gwpfieamwfigleam wtmm“ fifim fimga on rats

fiwflgmm fiam m fiyo

2 M pfiacmam m 6mm amm o

3 T 0 0m m qm mmafmw m mto m am m m m with chem icalmadam (m w ry mam tm m am emgp fl e a 3, m we have dammm mm fiem ’ m axim m m,m m mfmm)

Mm fi mfimm m figfm fim my mm Em M m am mm

m?m@ mmm firmw magmm afimm mm m mfi mefi bymm em fixm am a

3° CQBMbom mm z m nfim am Cfifiy Ewpfiam

«fie Pmbaficafimm am?fiwm d’gBo Fafiaoane gJo A ” , £73 0 9 C a EL, Bamw xm gJam and N o W e

5m m Age andm fim flmficfiy wfm t Mm ? m cfiei as m ewm m by vmmm e M d fifi fi comm aJo Gam ma m) pram )

Prepm gabyJoseph A , Eugene » giroNfim mber 5 9 1 958

Serial No 0 NHL-85

reteeto Twenty- one ofthese have totalbody density values

which are in areneonohfie range to, 95 to 1 ° These dataare being evaluated and compared to the other body compo

efitfion eetfimetee o

Direction (t Gnu-rent Reeenrch: A n integrated m eeefinxem ent pro

grnm nhteh tnctnnee eeteetee tneheee at hone sine and componitton e mhe contfinnedo hom e teehnfinnee ere provided th rom bcoonerntfion h fineeetthetofi

'e ontetde the Pnhfific Hentth Service

white othere nee etnnenreteenoroeennree noonin thie laboratoeyo Date nee nonth hfie foe eonnnnnteon with other phyefiolo

gfienfiw e hhfieh menhe eofifieetenon enhfieete oftheme etnoiee o

Moreover; enhfieete nhone efine (or eomnoefition»vnrfiee widelyfrom m ean eethen ettfihe eefieetenfor etnoyg nee experim entaland theeepentte h enteeem ente ot hone eom poettfion m y be

20 Pntfient henna None

hefittm oee one noeenee

hm htnntey hh Genny hefie heeeneeh tnetfitnte,hfihfioeh Cotteee 9 Yottee 89mm » Qhfio

he , hennfie heheneng”the hone henefity h eeenreh

and hentnntfion Center 9 Penneyteenfin etete Unfivefi' eate»Unfieeeefity Peek? Penneefieentn

De e Pent hereon90

The hone M netty heeeereh onehentnntton Context" 9 Pennehteenfie h ate UnfinexettypUnteeeetty m hp henneeteente

he , Sent h e heme» hennetnnent ot hteotetnegChfiengo hheahenfiSehoot» Chteem g Ettfinofie

Pnhtfienttoee nee heathen: h!one

Prennrenhe hethen h o Non-nth

N oeemhee 39 fihhh

Serial Na o a Mil-87

GERONTOLOGY M OB

Project Report

August 1 8 9 1 958 a» December 31 9 1 958

lo 3 0 Title s The role of eulfhydryl groupe in muecle actiono

be Principal InvestigatorsJecohJo BlueOther Inveetlgetore s

j a s“ s It 1 c N ote that the m eole p rotein m ele e which catalyz es

hydrolyele of adenoelne telphoephe te end of itc analogues

(M P? GTPQ G’PPg IT?) hee m ay eelfhydryl groom which are important in the

eneyh etie and lo the conteectlle proeeeeee e By the nee of certain SH weepblocking emote each as m echlorohw lheeeoete (Fee ) 9 it is possible

to accelerate end/ ox» inhibit the veto of hydrolyeie of ATP and its ane w

loguee o By deteiled steely of the hinetiee 9 it to hoped to deduce the acne

tributloo of the arom tlc ring to the binding of the oubetrete to the one

emetie hite a Such inform tloh will he of value not only in understandingthe h echenlom of ectioh of m ete e e en enem e g but m y aleo permit insight

into the nature of the tm neductloh of the ohemicel emery of A TP into he r »

chemical emerg e By cohmelng the effeet of mm on the contractile Pom

eponeee of the m em lm eyeteh when ATP, lTPg ete o ere need at substm tee g

it will be poeelhle to eoreele te the arithm etic effeete of SR groom and of

the m hetrete eroeetie ring with the hechehieel ohehgee o The contractile

effeete will he etudlefl on m eele (redol of veryleg complexity rangingfrom purified ectim nw’

oeie elmtw ee to glyeeeol teeetee peoe e fi beee o

Since it lo highly prohehle that the Sh y oope in the m ecle wotolo aotln

are intimately involvee to the coeteeotile proceee g it will eleo be necee=

m y to leveetlgete the effee te of.“ M on the ih tem ctiom between W m

M ectlo o

Eneyhe tic retee will he h ee eured by the Fiskeq iubherow

method m ahy pH m em eeente in unhuffered eolutlone o Sulfydryl y oupe

will be e etleeted by epecteophotometrlc procedure “ Gontm etile etudlee

will be done both ieetonicelly end leoeetrlcelly on glyeerol tree ted peoe e

fibere o Gheh gee in lehfi h wafer aggregation of ectoee'oelm in eolutioe

will he oxeelned primarily by light ece ttermg end eupeepeeoipite tion

etudie o o

home s Work will begin on thle peoj ect when the laborato-n

f iat ere reedyo

m “ ; “ H u m ” e 1 h .was Sulfydryl

groupe play on importent role lo the etm cture end wetloh of m ay pro n

tam e and any further knowledge of their role in the conversion of chemical

to mechanical ouerw will be of value in m m eeee e of blolowo

c 6; 6 Ji u _

fi t“

: 21 36 721

g in

Serial NO 0 : NHIw SB

January 1 9 1 958 c Dgeembar 31 9 1 958

lo 3 0 Title s Studies ou the gamm a-aim physiolow of sw eeoeneo :

Gamm alaria m gm w iamo

be Prim ipal Inw stigator s Em Lo Strehwr (1 0% of time )

(mm Inm atigaw mx Sam 's Crown 9 M ine Biological Labom =

torieze and Indiana Uniw mitywtthw Pollack » Baltimore City BowpitaleROW ?» Kahag Roelwfallw Institute

_

Gha na Hyttm baeh 9 Gw ame Institute

mmm w . Awmmm m Maleolm Gaye (logaf him )

go Pmm 'wa During Past h a ve Efim fiwz This raseam hg which was dew

aim to elucidate the em l m hanmm underlying» and the

pbyeiclogieal simifi amm e of fihe ayatm tflc ram-m ama of tha

hydrm tha @f the eolonial wax Gammm lariag was carr ied.

out at the Marine Biolagwal Labom toriaw9 bloom Hole 9 Masm =

chum tm o

n, m a M om that tln 0 1 6m M aths are gam ble to youngom g up to ism om et of regrw aiom in mm » cam city to ( 1 )

food (arkm ia) am (2) digewt and W rafi t food to the

Om ar £ 5? {Law of the eolemyo

E‘s: m 9 aim show that the m um mm tian of M P of W ra th

drew m um my dw fimg their am proaw s and m aches van:

flaking eoncemtm tiam (by extrapolatioa) at the time of ream :

M ama Thug it m y be that me li miting mate? for hydm th

m ciafiw cw is the available goal“

of purine nusleotideao

T um combined aw iam flaw s that W reaths do not undergo de=

gaam m tiom M um of a low of eapaeiw to function normallyo

Bentham“ may w ggw t m mm ania-6m m m y Mm an adaptive valufi

to am colony by forcing a radiat flhution of fm ing bodies wof great wt affieim y ( 1m m fi phm ) on a regular basiao

d o Dirm tion of M m fi Rm m h : We am attempting to culture thaw

m m in the laboratory in oft-

1M ? m inm wtigaw further the

mm hm imm of their growth aenww m e am regem ratiom eyele o

Serial B0 0 8 NHXG 93

m afia. Report

January 1 958 c » Bam mber 31 9 1 958

lo mo

53m bmmd mam a. infhwm d W azo o

h o Principal Inm stigafié m z Geozfm we

mmm Io Gm m rm

Other Im atflgafiom a No we Smockmm Jo Yiwglfi

mammal Amaimtam ae So 3 0 CM ?

c o Program Bumbag Pam. M IW Em m a“

mm data on 1 7-31 apt-um bythe thyroid gland in 9&0i aged M=9hwere have beenanbjm wd to magi

m im l @m lwis o No signifi cant, fl aw in

the 13pm 03 I by m M aid gland m x m m with mam as

mm M dim in mm Myanlz'zM ng blood wane found with im rw aingagao It 15 comclufleg m m, m thyroid gland of the M anny old

m m m M s available M Ma @m mgm tm m nom om m“

m m his

m ade tandem m aditiom M ardim ’

y (imam m a fi a .)

imam m of m mm zm m smm m to films 31 0a 0

do Dim em ozm of Su fism flam m ha work m we spasific pm j eet willIn eom lm d with t a m mratiom ai

‘

approm iata m m cn pta0

2° Patimnt Dam s flow

30 Collaborators s Baltimore city flagpitala providw space and all

he Publications am Am das

1 ° Watkins Do wa s me $1 89m of

Serial Hoo t NHI==914

«Maw 191 958 W W W ? 31 9 1 958

mm s magma m wfi m fi m mm of W lmm m agm a

Pgmmm l h m swgmw a flaw s—fl Lo sw am? (m w am )

mmm m m. gam m a gag 8m m Mam (m of Mm )

M a ma M ing Pam. m zm mmm g Cam idem bls aflm m m hawM en m m mm W e (1 ) m@ fl am m giw mam M um d

in M M mam a m g mm m aly M m 6m $0 be due to a

fiam lm iz h g effm fi w mM im d lim o (3) m m at-flaringcm is ch mly am $

5

0 W e metabolic wafiifi m of em m isc

rm h mmmm mam 31m m «am mim do

m mmm a»? 6m m fiam am m m a gam es.) M Mags m m m m am a

m m Dam s Riga; applfim mg

M bowsmom z m anure MW mapim lg

an catmm M id fim m‘

las Mm

M mm d by Sham Lo SW EAR ?

Gamma? 31 9 1 958

1 0

3

ac

SariufiN o NHI

GERONT OLOGY'

2238 A N C H

Jam m y 1 958 Dw embm " 1 958

”

mm ; Sm é fieg m the effect mi temmm fzww mmthe aging procesgoEL Baefiemm and m zymw

”‘agntmg

"gfiuflflego

Prfim fipaaKfivesfigatorz B , L, S hame? (5m

T eam im fiA ggigtm m g M , S mam e BM W (M%)

Program M fiimgm aTwam Mom-11m g B mm gg ether areas (sf

regem ch Emma deam armdm m y mmmmm them was M en; Wi le m a

ditiom fiwa marm this pm jefiitmag yeah Itm bam m e elem

that the filiwrmafiefim m am trim gfim pflg Mm mm m’ and rather

than imvegt a, Em gg fractficm of tim e mwag gam mago £11337wour effort on m are dim mfiy prom figmgmama mi m m m cho A il

mwgh merm afim ack; does m aappm r m m omma perm m m fi

m anage m MILMammow m vitmga, m ock» there is

a, daimte gem fitfim fifim a {cm b mmmaw thw mafl63am or to

m gtm mficm ofwe fimm fim m m fi ayatem m»

3 mm period aim ?

ewagm g if am expwm e fig mofi exfcrem e o

“mm deduced

from me mm matflam e m a, gra ter m m né flow at Emmam ne

cw m for m mm tim e after merm figaw k mm 1153 fig abgm ce g

Tm m mmm gmwfiy ggpm acmg um m agm am wa

dv Eim gtmm ai Cm m mfi Tm lum im gm m n m of

Euwww mm ma fig mm purifiw m sander to dam -mam

L Wham ? gm em flfiy repm fimefibflce and kfim fificaflfly Simmerefmctg em be M ama?mm m em m fim gygfiem o

2° Whefihw am em ym eg fim ofiw d a m demm yed m a, gtgpwimm am ar dugMgthexm afi dm aafim aitfirm

We Wm afi @m pfiz to m m fiaflflim {the Rummw cemfi em ym e g

fiufifiw m e g m auger m gfim m abgoamgfly m fifiorm m iefl afi

Wham Egmmfic behavmm (cm be mw gmm fi Wm; gm e o

m am Daygz N ome

C aaflamm torgz Bafltim om {M y Hawaiians

Serial N o NHL-9?

GER ONT OLOGY BR A NCH

Project R epere

T ittie s Verbefleerform eee ee e, fieeetfiee efi egumgo

Geeks The eteey of veeeenemailknee been eem eenm eegfiecfied

refiefive fie gamer ekwege aeo m eme e fie wee hope thatweeegfleeemanbe m oefifi ee; eee m: ie w e belief that the etefly of

verbenm eme-m em e Wm {Leee to eflgem eeet ffieeirmge regexdiegage emereeeee fire a m fieey efpeyefiwflegfieeflpreeeeeee o We

me me, therefore” to eeeely the eefietfieeempe of eefieeted perm »

etezre veebefieeeeefioa' eo ageing» em {to eveeteeuy z eliate meee

peeem efiem m etifinee peyefimflegfieeflpeeeeeeee emeflnee comm em a

cetme eeee eee end with the egee Two peeem eteee oi eeeeefi

behevfier ew e been; eefieefiee fee peemm fieery eeeeyg eeeodmgeefi

'

fififiyg em m eeefiege ef ege e eefleveee eeeeepfize a

fie W fitfineeepeefz fie eeeeefiegtee Eeflfleemgeyeefiheefle me bemeted: Eem ee eee twee efi

’

eeeeefiegfieeeeeee ee e feeefifiee

ofwe“M ile hypemeefie fie efim pfiy em emeeeiee eff whet Rhee

eeee N eed with other eem e w e peefiwem eeee o We e fieh fie

eee e meee fi efiiflegge geeem fiiee (to veereefipeefieemeeee o

no m e eeepeeitto m eeefiege tee e wes/7 em be eeeerfiptfive;fie e , we em . efim ezy eeeeeeee dififieeeeeee fie eeeeept m eee iege

ehfieh eefietm e eige fifi eeet eeeeee bette eee egge groepe o

me m e: Hypezeeeie L The eragfiee‘

am eme fer aeeeiege mfiypefi eeie g time to ewefl eeee m anewe yew » has b

e en

chewed efieee the neefl: eeeeefieeeeeem the feflflee fiee emeeitoTee eee wewe Ffieeeflneeeeem eeee deem ed?elmwe arememe e we mm meme (1 ) m y the ew e (gram w to 25 wee

m e (2) very fie degree fie which theywexemmete m eemaegeaEegnfiee Mem e eeeeeeeee g The marten-fleafie eeeeeeee beck

w the fieveeefigeiter em eeeeeeee em; e name for eealyeie o Exa

w ee ofm isuse m e m e be M fieeelem m eee eredn T ech n

Mem e ewe being eevefiepeeflee m eeeeriegMm e eoeeem ed by

eeeeee (tim e befae eee waffle ) eee F937weweueetfiee (Mm e eaten »

in e eedeh Em eme end mim e Wififlbe reflefie‘

fi m age ,

130 T he eeeeeefifie dfififee‘

eemefi em be emefleyee to m eeem e the

m eemege of eeefiefie eeeeeefie o Em mpfiee efi peeefieue eeeeeeee

ere Dfiefl fip GLD M y A D ©LES©ENT 9 em , C oeeepte need

M be weed Earem me W em fiee e emailfrom efleeeeeieee bette eee

the pfl eefipefi fieeeefigeeoee o T heee eeeeeeee em be w eed rm

eevee n eefiefi; eeeflee defined by edgeetwe m epeeefitee; e a go

genie-beep eetrfie e

“

m e m e e ite eee m weeeo T heee eeefiee e m

L.

SarinaNo NHL-98

GERON’

IOLOGY BRA NCH

Project Ream

Jam m ywee M em e wee

ao

“flue : Reeeuee flame m e efleeevneeeemenegemmfie careefiefies

as fieeeefieee of egfiego

me 5L. xefieeemxee fieeggs ehm gee mwe enemafieeeeeeey of

the eaeettreeeeegetmeflm m Gee weemeg M eme fie deeiv

ed from me ebeewame‘

ee (amfine Rateeeitm ’e em (1 ) gimme

reeetfimm tim e fimereeeee w e age » w e (2)me m ew fireqmemeyofthe efipme 3't cfime efieetm eeeepleefiegm m deexeeeee

withwe We hypotheeaz ewe the reeeefiee c efim e emerem ee

bem eee we g am e em be eeeeeefieeflees in pert» by emfeeeeeee m we efipfie freeeeeefiee obeew ed bem eem the m o

gm epe o We aaee N ee to efiwwwe we fieeeieemny repemedm iebfififity m efimpfle e é eefi ee fume, efeeewee fie the ew e

ieelflmdeeflever a.m eme efi‘

m m » fie e fim efi ee ofi the efipmteeeeeeegpeeeeeaefime eime an?gem em m o

Methods A efimpfie eeeetfiee a tfim e peoeeefim eg m which the3m m? eeepeeee mm,

e em mm wmflefi'

afie emfiegmefined atwe

term ne whenwe r]:as. ewe m fie peeeeefip fie weedo“M e w e

jeceeeepemele fieweM eeeeeeeeee of me m mby M ermgaw eed elem m m mwe eemee efimeme he te eem e fimeflee

eee fame fieege eeey tee m m for

efi elem fie efiee emefieee eefiegee m em e? eeee ee the emmewflee , Ceevee effreeefifiee sam e we , BBQfireeeemey ere pkefitefiflee eeefmeebgeefio

be Pemefipefifi weetflgem re: Wemezr W0 Sw e fiflfie (2/ 5 Mm e)m ergeJSwea (We tam e)Mem e D o Demfiefifi W e time)

T eefimfieeflM efieeeeee : M ike m eme (fieafiwfifim e eemm er 1 958»{rime efieee Septem ber)

Peeg eee Derfimg PageTwelve M M ° Thee peefieefiwee nam ed

L, wee A reeeeaemwfiimme eppm emg m am fie com

m ime menEE G eeeommgm Deere deefigeed and ceeefim eeed;

gram me bee beem made fee We ewe? of ehefiee me weanem

eimme m ammalMm e »

SerialN o RBI- 99

GERONT OLOGY BR A N CH

Project Report

heather? whh o Beeem hee we

to e o T ttte : The theetteh at m etet thee he eheemette eeeetwhe o

he Petheteetthveetteetee : hehthet Lo htehheee (M2 tame)

T eehhteenheeteteeeez eaten Oehehe W e the e)

Gheettvee: T o th rive ht eh heeeeeteeeteg at the theehehtem by ehtehH eme hexttetpete ah ete yheatte eeeettehe9 ehe etee to heteem the

why dttteeeht m eted tame th e eeeeeeee tee ehteeeet em em ette preeeeeee o

etteehm eet to the eeheteete 9 he (2) tt meet mehet e tth the em m e en tete,

meetth e ettezrthe eeeehhew hate teetteey eteeethee at the wh en heeehethe the eettve eehtteheettee o it e he mm ed thet9 ehehevee heheh e (t)eppttee 9 the m etet ehehte he eehehte at hethe

'thgeheet e h eh e ehzymatie

reeetteh ; tetteee at he h eh c eheymette eeeettee 9 theereetm e thane favor

eeheme teh

Sheetetty eeteetee em m ette meheteetee aee teeetee‘

le tth m etet fich e

m eet m eme eeehtttehe9 ehhthe ehehehe ht the m eteto ehhsteete thteeeettehe heteetee he hhheteet eeeehte9 e ohe 9 eeeeh

'ephetem etry

The teteeeettee h the metete mm the ehehhee heetete te etehteeWmeet ee eteeetee teehetheee 9 eheh he etteeeeetettegetm

eth er EthethgeH The eeeteee ehe eeeetteee eeeetteee heave heem eeteetee

"

em mh eh eheehthtte eeeettehe the aeeemhtteh were etm ttee m etech a

the teehethemmettehe 9 het reehhf’e eht

‘

teeeht m etet thee, h e 9 memeehtm

heeltree (fifth eeeheetteeh'o The eehetxfi ee ct heth eeaettehe have been

theme to heeeeee m etet cemeteeee at“

m ettle etehtttttee9 het evihehee teee eeh m eheymette eeeetteh hae heeh ehtetheh eerie fee the thteeeetteh at

th ee e tth the eeehtteee ehheteetee o

he;ereeee him -

z “ 0 56 515 11 ah the hettetttee at eethntee eh

eym ee o Steetee ht em heette meehehtem e ere 9 theeeteee 9 at th eeemehtet

thteeeet te the heegeam et the Get eetetegy hehheh

hteeetteh at Cm eehtheeeeeeh The hteettee at the eeaetteh at tree (me 2 e9 ew e are a eetteete withhe etehtee9 ehe the t eteh com

paeee wtth the teteh thetna m eeheew eh toe the eeoeiteee reeettome

Physialogy

3° Be thesda“ Eé rylwd

Efifi a NIH

Individaal Praj e@fi ReyawtCafiemdar Yfiar E958

Prajaam Tfitfie s fifiasmremafit wf'

vemtricuaar Mmsefle Fiber Length

Primeipafi Jo fifitehenfiq Ro 3° Lindeno

0th®r Imwestfigat s : N®m@ f®®hmicifimsa Nome

C mmm tmgm m M m

Mam years (caflemdfir year E95®33 Pfififiemt Dags a Name

mammam a 94w m m

Tfie Cashmy n®v@m primafipfi@ has b@@m- mmdfi§ i@d as to

it sws@ep®imae wfl'

mmfiswrfimg emamgss fin wemfiz ifimnar Emacae

ffibex E®mg$ho The @@msisfis ®f mmm amms , am@ of wfifickth@

'

shafit @E Q flww fimaxtfia auerazowqme pwtamtiemetero

®f this p@t@mfifi@metex is awfitfiwwemsly reewxded thrwwgfiS@&h@ra 0; C o e@mpflim@ ampfififierq Tha arms ®f the l®wer are

samfi fimma tfim p®fimfis wa tag fi®ft amd cwawges £3dfistamae EQQWQQm twa paimts ta@r@by It is

assmaed that ehamges im fihig distam@@ ehawges infiih®t gewgfiho Ta fiate o th® fi®ver s hefim aged in mfima dogs q

Pr@p@s@d a®utse ®fi fih® prfimafipafi infi®rest in this

fiflan eamma fis .a@ ha ©bfi® ta f®fifl®m in diastafiic fiberfiemgfihg fib®$ fi®wgfiho amd ehamge im euntramti@m

and %&th th@ othex sfiflmfitawewmsfly reearded

it is h®y®d ta b@ ahfle fia dfifiQffififi® chamg®s a fif anyo fin eada

diasa®nfic dfiafiemsibfifififiy mmd@r am@ imfiflm®me® af hwt& cardiac

symmfi thetfiwawd fifififim£@3fi@m whatm®z amy each engages ar@

due either fifi® fi@m@fiffi@ @E the mervw seimmlatiom a?

efiamg@s in dmr@ti@m af g@@fle ©E m@aa ama fiWQEfiQQlfiy to

examflme awre ia t fififl th® endc

diastenfie ffib®r and wmptfiamo

Pgrt B imcnwdefls N0

Sexism. wo o

”11 4 3-05

fiwailabne fi® it efferent pathways by means af which

mamfi tiaalny mmmipmnate the t9 33ti®fishipi betmeen filiimgmwe and weatriewfiar str@@e awake

s fiher exggrfimemms'

w@m@ angm dame t® ascertain whetfierafferwmt pafimmays ara exem@fi@@fi by em&m@@s in carotid siflwg

immnafifi®m @f the car@tfid afimms m@?W@8 pradwced

mm m nmm timw absewed m en with:

ympathgtfia stimmiatfiam ar fimfltiatimg stiwmlaticmg

£3 0 a shift of th@ ventricular fwm©tfl®m curve to the right o

tfiwm @f the sar@$fid ginms gm®ssmra mad the effect o

flag mm@ earwtid pmesswre shiftad am@ wawtrfiewlaz fumetfi®mfio awe Refit , fihe abserv@d w@r@ In

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Physiofiogy

3 ° Bé mhesdab Qé ryflamd

ladividmafi Qwoj ecfi wepowtCsfiemdar Year 3938

Eraj eet Tififlea The Passibie PhysfioflogfieanSigmflffieamce ofWaswdfiflator Substfinces in Qamé m Urine

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l o Aqueous solutions of neutral salts which have large effects

on the configurational properties of poly=l=proline and on the proline

residues in glycine wproline copolymers also have profound effects

on the properties of col lagen and ge latin suggesting that the peptide

configuration about proline wproline bonds in these proteins is

altered in these solvents o

2° The eiiect of temperature on the optical rotatory properties

of ge latin paralle ls that observed for the glycine e

proline copolymers o

Since glycine residues to not exhibit optical rotationy the changes in

rotation observed must be re lated to the configuration of the prolineresidues about the peptide bonds o It is suggested that at low tempera

tures the transconiiguration oi proliue=proline bonds is predominant o

A t higher temperatures rotation of one proline with respect to its

neighbor can occur and this e liminates the structural patternestablished at low temperatures o

do From the mark with glycine proline copolymers it appears

that the enzyme col lagenase requires the specific sequence a ,

X=glycine e proline a » in a polypeptide chain in order for hydrolysisto take place s Cleavage of the chain occurs between the nonspecific

amino acids X9 and glycine o This information may be of value in

future studies on other as yet uninnestigated proteins in which this

particular sequence may occur o For exampleg the anterior pituitaryhormone pu rse is known from structure mark to possess this sequence n

4 0 An examination of the hydro lysis of ge latin by col lagenasereveals that at temperatures below 25

° the normally random polypeptidechain of ge latin fo lds into a much more organiz ed structure “ The

special configuration of the ge latin chain be lon 25° apparently isclose ly re lated to the ge lation process and to the formation of the

unique structural pattern associated nith col lageno It is be lievedthat the structural changes at loo temperature resul ts from Cisa trans

isomeriaations at the proline e prollne or proline m hydronyproline

peptide bonds in the gelatin chaino

gim f incense $0 Heart Research z This sort should help in

clarifying the properties of certain proteins containing unusuallyhigh amounts at proline and hydronyproline o Co l lagen and casein are

example s oi prote ins whose structural properties appear to be primarilyre lated to the proline residues o Collagen9 of course y is the most

important structural protein in the bodyo

Sef ie i mo oNfilw lll

PHS= w NIZH

Individwei Prej ee t ReperiCalendar Year 1 953

Jenei sg Publieeiiems

Pubiieeiiens ether than abstracts irem this proj ect

We F o Herringten and Michael Se leg Studies on the Structure

( 1 958?

We F 0 Herringieng Effect of Neutral Salts on the Structure of

Ceilegen and Ge latin? Nature 9'

1 81 9 997c 998

Serial No oNflm° 1 1 2

20 About 20 per cent of the myosin molecule appears to be

amorphous”or possesses a much looser hydrogenwbonded fabric than

the remaining 89 or cent o The”amorphous

"0

areas of myosin appear to

occur at about 1 d and the length of the myosin rod as j udged bythe siz e of the high molecular weight degradation products whichare rapidly formed on enzymatic attach e

gflhnugfiuunumulunihmmm_hesearch3 This norh is aimed at a better

understanding of the nepects of structure which make a protein

fibrous in character ° Fibrous proteins9 inc luding fibrinogen9 the

collagens and the muscle proteins9 appear from the present work to

share many similar features and the systematic study of these proteins

along the lines described above should help to unearth any common

structural patterns °

l o Studies of the secondary structure of fibrous proteins

utilising proteolytic enaymes will be extended to other fibrous

proteins such as iibrinogeno

2 0 Model polypeptide chains such as oxidised ribonuclease and

simple synthetic copolymers mil l be examined e

Part B included Yes i

l o Laboratory of Ce llular Physiologyand Me tabol ism

3 0 Be thesdap Maryland

PUBLIC HEAL'

lLl SERVICE m » M TIONAL INSTITUTES OF HEALTH

Indiuidual Proj ect ReportCalendar Year l958

Pro ject T itle : Energy transfer associated with e lectron transport

Principal lnue stigators u; Wayne KielleyJo Ramsey BronhOther investigator 3 Lisa Barnett ( technical)

Clarence Edwards isummerl ( technical)

Man Years (Calendar year nasalTotal s l

Professionals l

Technical: d/d

in most aerobic ce l ls the mayor pathway for disposal of e lectronsarising from oxidation reactions leads to oxygen through a system of

closely integrated protein and lipid elements forming the membranes

oi the mitochondrial components of the ce ll o integrated into this

l ipoprotein system is a mechanism for transferring the energy auailw

able in the sequence of electron transfers into the form of high

energy“

phosphate bondsg presiding a maj or source of energy for

aerobic organisms o

in this l ipoprotein system the sequence of electron transfers

is incomplete ly hnonng ten of the indiridual elements have been

separated and characterised9 the nature of the intermediate in energytransfer is unknown and me can only speculate as to the function of

the phospholipid components o These problems and the questions of

physiological control mechanisms form the broad obj ectires of this program o

Progress in understanding the mechanism at the oxidatiue

phoSphorylaticn has been great ly facilitated by the derelopment in

this and other laboratories of procedures for fragmenting mitochondria

to yie ld much smaller units capable oi carrying out the integrated

oridatire phosphorylation reactions free of interfering and complicatingreactions present in intact mitochondria » Precious reports have dealt

with the properties of these submitochondrial particlesg and association

of two exchange reactions with the phosphorylation process » the derelop m

meat of a reaction scheme and point of action of dinitrophenol in

PEIS M NIII

Individual Proj ect ReportCalendar Year 1 958 Serial No t

NHI° 1 1 3

Part B : HonorsP Awards ? and Publications

Enhllgn tggnga (other than abstracts from this projec t )

no N , Kie iley andJo R 0 Bi onic9 Oxidative PhoSphoi ylation and A ssociatedReactions in Mitochondrial Fragments Obtained by Sonic Vibration o In

Proceedings of the International Symposion on Enzyme Chemistry9 Tokyoand Kyoto, 1 9579 A cademic Pnessy New

‘

it’

onizf9 1 958 0

J° R n Baonkg Some Effects of Thynoxin on Oxidatine Phosphotylation

in Snbmitochondriai Particles and Intact Mitochondria? Binfihimo fiiflflhgaa

Afiifla 21 9 (EQSBDQ

J o R Bi onk and N 0 W0 Kielley9 Evidence for the Point of Action of

29 4 =Dinitcopnenoi on ATPase 9 ATPa 32P Exchange and PhoSpnorylation9

.BiQt itLa BiQPM Qg é Qioo .239 369 5 377 ( 1 958) 0

we We Kie iiey andJo R 0 Bi onic9 Oxidative Phosohoryiation in MitochondrialFragments Obtained by Sonic Vibration» ifi m aioiaggmanp g ZQQQ 521 a 533 ( 1 955 i n

Serial No NHI- llh

l n Laboratory of Cellular”

Physiology2 o

3 0 Be the sda» Maryland

PUBLIC ElEALTIl SERVICE w NATIONAL INSTITUTES OF IE A LTH

Individual Proj ec t ReportCalendar Year l958

Pro ject Title : Biochemistry of muscular contraction

_irirsiralw lnrsatiwator s We Wayne Kiellet

gollaboratog William i n BarringtonWilliam d o Dreyer

ggher Investigator Lisa Barne tt itechnicall

Man Years (Calendar year l958lTotal:Professionalz 3f1rTechnical: iii

Objectives t I t was observed some time ago that the ATPase

activity of myosin9 the protein unit of the contractile mechanismg exhibitsa biphasic response to titration of the SB groups? with a be d fold stimulam

t ion of ATPase activity occurring when approximately l/2 of the SH groups

are titrated, further titration leading event ually to comple te inhibition s

The question exists whe ther the two phases of this response are due to

titration of specific groups or to statistical titration of the groups

involved in each ac tive center o I t was planned to approach the problem

using differential labeling of the groups with radioactive SH reagentsfollowed by tryptic digestion and application oi the

”fingerprint” technique

to identity the cysteine containing peptides o Howeverg interpre tation of

the results would depend on whe ther the large myosin h molecule consists of

a single or several peptide chains and ii several9 whe ther they are identical o

Therefore ? it was decided to approach this question firstg using concentratedguanidine hydrochloride to disrupt all of the secondary and tertiarystructure s leaving only the primary covalent bonds of the peptide chain oExamination in the ultra centrifuge should then establish the siz es of the

basic units o hyosin A is a rather large molecule (400 9 000 molecular weight )but offers a unique opportunity to establish some de tails of fine structure «

it is a very asymme tric molecule ( 1 600 fingstrom units long and 21 B in

diame ter) and the number of ways of arranging a single or several pept idechains in this shape are very restric ted”

1 1 1 1 1 N1 0NHI ° 1 1 5

1 0 1 1 1 1 1 1 1 0 1 1 0 1 61 1 1 1 1 1 1 Physi®1 ®g2°

3 ° 81 1 1 1 1 1 1 9 M1 1 yland

[355 1 4 131 21

1 1 d1 1 1 dual P1 aj ecm Rep®1 tCalendar Year 1 958

Emgjegg. Tg1l@ : 01 the 1 1 1 1 1 9 1 1 1 1 1 selfu 1 1 31 1 p1 1 0 1 p1 1 1 ®menan

6 1 1 1 1 1 1 1 4 0

figfi 1 1 1 1 1 (Cslendar year 1 1 58) Eggigfit figggz N©ne

9 1 11 1 1 1 1 1 3 1 1 1w1 1 1 1 1 1 1 Em91 1 1 1 1 3 6 1 1 1 1 1 1 1 4 1 8 1 1 1 m1 st widely 1 1 1 1

isozape 1 1 b1 © 1 1 g1 ©1 1w

1 e3 1 1 1 1 1 0 31 1 1 1 1 1 1 1 the 1 1 1 energy 1 1 1 1 s 1 1 1 1 1 1 1 1 9

1 1 1 1 5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 by 1 1 1 1 1 1 1 1 1

1 1 31 1 1 1 1 1 1 1 1 1 1 1 1 1 1 91 1 1 1 1 1 1 1 1 1 1 0 1 1 1 5 1 1 1 1 1 1 1 1 1 1 1 be 1 1 1 1 1 1 1 1 1 1 1 1 1

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 8 1 1 1 1 1 1 1 1 1 9 1 1 0 P1 5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 11 6 1 1

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 g1 1 1 1 1 1 1 9 1 1 0 1 1 1 1 1 been 1 1 1 1 1 1 1 6 1 1 1 1 1 5

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 hype1 b®1 1 1 0 1 1 1 1 1 1 1 1 1 1 1 1

5 1 91 1 1 1 6 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 9 is 1 1 1 1 by p1 1 1 t 1 1 g 1 1 1 0 1 1 1 1 6 1 1 1 1 1 1 1 1 1 1 9 1

1 1 1 1 1 91 1 1 1 1 1 g1 aph 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 g1 1 1 1 1 1 1 1 1 9

1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1g1 1 y 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Eamge 1 1

we 1 g1 1 s 1 1 6 1 1 1 1 1 1 1 6 0 1 1 1 8 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 and 1 1 5 1 1 1 1 1 1 1 1 1 1 1 app1 y1 1 many 1 1 1 1 6 1 1 1 1 1 1 1 1 1 1 1 g 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 9 1 1 1 1 1 1 1 1 1 type ©1 untingequipment and 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 has beenfound 1 1 1 1 1 1 1 1 s1g1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1 1 1 1 1 1 1 1 1 ©b1 1 1 1 1 1 1 1 1 1 1 1 1 1

advantages 1 1 the 1 1 81 1 1 1 p1 1 1 1 1 1 1 1 1 1 md grea1 1 1'

1 ccuracy 1 1 1 1 1 1 1 1

1 1 1 1 1 1 1 1 1 » 6 1 1 1 1 1 1 1 1 1 1 1 8 1 1 1 1 1 8 1 1 3 1 1 1 1 1 5 1 1 1 5 are inwalved o The 1 1 1 0 1 6 1 1 1 1 1

1 1 1 1 1 1 1 1 1 1 1 1 1 5 1 1 1 1 the 1 5 3 1 mp 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 absorb1 1 g 3 1 1 1

always 1 1 3 1 1 1 1 1 1 1 3 1 1 1 1 p1 1 0 1 1 1 1 1 1 1 1 1 mp1 1 g1 1 g 1 1 1 1 1 1 1 1 1 0

The hype1 1 1 1 1 c 1 1 1 1 1 m1 1 1 9 1 1 1 1 1 1 1 1 0 1 hands 1 s a 1 1 1 1 6 1 1 1 1 1 6 1 1 the 1 1 1 1 1 1 1 1

1 1 1 1 1 the 1 1 1 1 1 1 1 1 1 each 1 1 y6 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 A m 1 1 1 1 1 1 5 1 1 1 has

1 1 1 1 d®1 1 1 1 d 1 1 1 1 1 1 1 5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 as 1 1 1 1 1 1 1 1 1 1 1 we 1 ght 1 1

sample and 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0

g?1 has 1 9 1 1 1 1 1 1 d 1 1 1 1 the game 1 11 1 1 1 1 1 1 1 seems 1 1 1 1 1 1 1 1 1 1

ur a fifir and 1 1 1 1 1 1 5 1 1 1 1 1 1 1 1 1 1 1 1 5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 81 1 1 1 1 1 1 g©1 y o(D

U? 1 1 1

1 1 1

Serial Ne eNfll‘ llb

lo Labet atory of Cel lular Physiologyand Me tabol ism

2°

3 ° Eetiesde 9 denyiend

Pisc a difl

Indieidnei Ptej eet depentCeiendet tent l958

gggiggtglitles A study ed the neenenien ei pnetein biesynthesis in

the hen evident

Emdnddnndh innefiiignine : Rienend we Hendlet

Men teens (calender yene idfidl imflggnggjgymnz Nene

Tetel:

Pneiessienei :

dnjeetine a tee neenenien ei pnetein eieeyntnesis is ene ed the

nest impet tnnt eneee in n_ieenenistty enent etien the Benet is intnn o

it is tne pneneee ei tnie went te sindy the entie teeetiens ei penteie

biesyntneeis in e tiesne highly speeieiieed inn petietning this fnnctien9the evidnet ei the laying lieno

Tne existence ed en intermediate

stage betweenfree enine eeid end penteie nee indiented by the fe line “

inn ebsennntinno Free enine eeld deee net beeene ineennet eted inte

pnetein in e distnpted dell pnendnetien nineteee9 ii the cells ate

lebeied nniet te diennntienp t edienetieity een entet the ni ete teit em eene iebeied pneeneset in the telie o in ettennt nee mede to

leeete the eennee ed this nedieeetit ity as me ii es in enenide other

eellnlen eenetitnente inn e nest inle pneenteet inle t Tee entite tissne

nee examined in diffenent etenieel tenetiensg fee the rate t i eeennnie tingnedieeetite amine eeide9 the nteeinte letei ei eeennnletieng and the

dynamic state ei the nedieeetit ity in neletien tn the rate 0 5 idcdrpdfm

etien ei the enine eeids inte pneteino Preliminary steps were taken

in ehenectet iz ing the ehenit el netnne ed the binding ef amine acids in

tneee“neno pt etein

00

it eetiene o Tne neenits indicate that9 eentt ety te

expectetienss the nneleie acid it eetien dees net behave as medid be

expected fee a te le ef intennediete ent eiem ei amine eeids ° The iipid

fret tieng on the ether hand» denenstneted kine tic betat ter with

neiet enee tn amine eeid intake end tntneten9 entiteiy eensistent with

the pessibility ed its pieting the nele ed intennediete cent iet fee

the amine acids o This idea nee int imen stt engtnened by the ebset tetien

tnet egente etimnletety tn lipid synthesis stimulated emine acid inm

eerperetien and egente deteinentni te iipid sttnetnt e steengiy inhibited

amine eeid ineetpenetieno The peseinility ei lipids being intelted in

pt etein synthesis nee nitnet te been igneted o t enet 9 en eytelegiee i

and thetnedynenie gnennds tne pessinility eenid eiiee distinct

advantages o

emo Publications

than ebstrasts free this

Rishaid Wo fiendiee 9 Possible Involvement of Lipids in ProteinSynthesis, figiegge, tees 1 43m 1 44 ( 1 95830

Rieheed £ 0 fie ndiesg Passage of Radioactivity between Protein

Fractions of eJen Ovioect Iioeogeeste 9Jo Biogo Chems g ‘gggg553=56i ( 1 95730

Serial No oNHI ° 1 1 7

been located in one of the several prote in fractions o The amino

acid sequence of this protein is be lieved to be altered in a different

way in each of a number of close ly l inked mutants of the viros o The

icons of each of these mutations on a gene tic map has been determined

by Br a George Streisinger o De termination of the precise nature of

the changes in protein structure resulting from such a series of

mapped genetic mutations should ul timately lead to a be tter under ”

standing of the may in which the00

genes00function in the control of

protein synthesis °

20 Deve lopment of methods for the study of protein structure

In order to male amino acid sequence determinations possible in

large protein moleculess such as the bacterial virus serum blockingantigeny a re latively simple method has been developed which permits

the separation of up to one hundred peptides obtained from enzymatic

digests of proteins o the me thod? based on tmowdimensional electro a

phonesis and chromatography9 seems to be not hing h el l and is now beingapplied to the gene tic problema

themmmmmmnums_a_lysonmge8 Certain bacterial viruses contain an enzyme

capable of degrading host ce l l malls o this enayme is also found in

tree form in lysates of ce l ls infected with the rinneg but is not

present in lysates of uninfected cells o a convenient assay system has

been devised for this enzyme and a procedure has been deve loped which

permits isolation of the enzyme in what appears to be nearly pure torm a

The enzyme was previously considered to be proteolytic sinceg amongother compoundsg amino acids are re leased by its action° however9studies oi the purified enzyme indicate that it is a lysocyme rather

than a proteolytic enzyme s The iolloning list includes some of the

interesting features of this bacterial virus enayme

o n bacterial virus lysonyme appears to be synthesiz ed under

the genetic control of the virus o Since the enzyme seems to be

re lative ly low in molecular we ight it may ultimate ly prove to be

particularly useful for the study at genetic control of protein

synthesis o

be There are certain similarities betneen this enayme and the

one believed to be present in a number at mammalian viruses

(e ggs iniluenaalo A study of this type of enzyme should shed

considerable light on the mechanism of viral infectiono

c ° A comparison of the structure at this enzyme and lysosyme

from other organisms (e gg° other viruses9 egg white lysoayme 9 etc ol

should provide information about the species differences of

proteins nith similar activities o

do The enzyme lyses d o call igram negativel under conditions

where egg nhite lysosyme is ineffectual as a bactericidal agent °

e a it is one oi the very ten enzymes found in viruses o

Serial N9 0Nfllm ll8

Methgds :

Preliminary results tuuteete a ueueuet ueleculet weight of 1 009000

(plus he mihes e ouo fut eyestua Simee the literature value

tut the uuleculuu uetpht et upestu us determiued by conventional

methuds is 4009 0009 this uesutt suggests that uyestu as it exists

in setuttuu ts eeupuseuut-S u h fiueutteet suhuuits ®f sheut 1 009 060 Mowo

eeeh o It is upped that it h ilt he pessthte tu detetutue the uuuhet ef

subunits with uteutee pteetsteu hp‘

eeueleptug the uethud fuetheuo A

ueuuueu ueleuulut uefipht ht ®oao Shgfifih use ehteiued fut actiu

( thtet etut e tulue th te hhghfihbo Thepeuyustu puue u uuuuuer value

eluse tu the uepueted uetue tehgfihhho These peeltutuety tesults else

indicate thet9 eeuteeep te eeuteiu puepesefls fuuud is the liteustute geeeh hi the uhsue peeteius is e ututtuet eutttyo

A ttempts hsue eeee uede tu study the peptides euutuiutuu chemteettyhluehed SH gteups o Hhueeeug flue te teehuteul diffteutttes he

results see stutluhte ut pteseut o

There is he immediate stuntfieuueets heuet teseuteh9 theuuh uhee exteusfite hueuteuge ut the peetetus

tutelted tu uuseutsu eeuteeettuu use he hf eueutust tuput tsuee a

Peepeseu Cuugse ef Preteet z the sesuhts tudieeteu sheue see iucuuplete

sue shuule theeetete he eeuttteeu eud euteudeuhp further studies o

Seueuel pessihle ueeus et fiupuet fiug the ue thuus used sheuluhe exeutued q

Since the Sh gteups ef the upestu uuteeufle appese ts he implicated

tu its eututytie uetiufityg they eheutuhe eueutueu tu suue detailo

Ideuttfteetteu hf Sh eeutetutup pe

ptides flu preteetytte digests st

upeste sheuluhe pesefihle e

PART B thetuued me x

Serial No .Nil-1 4 22

l .Lebore tory of Cellular Physiolomrand Metabolism

Individeel Proj ect Report

PM A

Peoj eet T itle Sfieflie e an th e eem ewre of lyeeeym

Prieeipel have efiigetor : Arnold M M iss

M e Yeer e (Calender yeer Patients new : Nome

mgeetive : T0 ieolete seed identify, by tee -dim m ione l peper

m em ephy em eleeteephere eie the pepeidee free e tryptic di m

Mem eee w leged : A new te chnique fer the peeeiee eepere o

m en of peptides kne e evolved. M enage were in m m depw teemt em

eleeehere 0 M e eppliee

’

m li‘ey in the detezfmieeitiem of the am ino acid

segmentee of e. preeem enz yme of ,000 m leeuler we ight file being in

veefgigeted e e e M el fer future week 0 e m ine lyeeeyme is re

M m e of .peptide e o

“Maxie mixture fie reeelved mm individual peptide

spots by ehreee teg ephy, fem-wed. by electroyhereeie on le rge sheet s

of fi lter paper o The m ememg pattern ef peptides is celle d e

”fi ngerprint

"

end. ale (mem e cem eterietic of m m prem is e It be e

be en m ee'

ible e‘

fiz m ie“

time to M emefify e ight, of the eeven'eeen pep

tidee expeeeed in mm eigeetiom mixture

week fie pert of the

me;

of the etm eture 9 bioeynm eeie em M etion ef m e te in m le cule e,

m im w em ee be vmg emeyezem e prepeeeie e o The were in gene ral

is ef e. highly he eia em m eeeetieel m m with e‘

evioue beermge on

eieeue me‘é ebeliem in genezeel o

Pffi oeee jeem ee e? Progeet : Further work is fiireete d to

em eueeeeeful identificat ion of the entire eemplemem: of peptide s

free the em m e dige et of lyee eyee o M e is expe cted to denm netrate

the usefulneee of the fi ngeepx’in

'e

“m em e for em lyeie were em}. per

m e we epplicet ien to the m ay problem in the etudy of

prote in

etrue‘

eure ema it e genetie determinefi en new be ing emailed in thie

m ewB inelude d

Sex 0 No 0NHI@ 1 23

(Maj or fim ngs conhinned )

conolnied that the ehylonflcrone isolated from «loge and hum ane contain the

high-denem y M yoprote in prote in 0

d . The eolnhilflty propertie e of the B prote in free both dog end.

hnnnn ohyloniorone ere 8m m The fi ngerprint"

gen om e of theee pro

teins ere completely different from the higme deneity lim pro’ce in protein on

well e e fine plnenn loe m deneity (Sf 0 - 20 ) protein in hnnene and done , This

eviflence p eonpleo. wieh previone fm nee e m honeys;Johannes oon‘

hoine A lipo

pro'

heine nith N-eerninel eerine (the eee e e e then found. for the B prote in in

lum en ehylomfierone ) enege et that ehie peofsein (or M poproteinfle fundame ntal

for m e fez-renepom

2 o e o In vivo otndie e on ineorporem on of labeled enino onion

into the profie fine of the ehylonfienone M ohamed fem . dog W k: during the

eheorphion of fee neemnetrehe thee them ie e te pid inerenee in the ene

eifi e eeewm ie e of the two m jen ehylonnenon prote ine . The eneeifi c activity

of the A prote in in ohylonnerone in ideneioel he th e epe eif'

io activity of the

higho de neihy li popro‘eein in lymph huh eoneinezeehly higher then then found in

the ple em high e deneifizy M poprotem m e oeion . while W oes than the ple eee.

high o oenefiw M popro‘hein in not fin equilibrium wifeh the ehylonioron A prote in

or the W h high s-nonei‘

hy M poproeein o

h o in were etnnie e on {fine fineorpore‘

hion of leheled w ine m m

in ehylm ioeon proheino hy inheehine l nnoooel ce llo indicate then the inc

neem nel oeu n ere oenehle of em ehe eiefing proheine re eena g the A one B

proteins of ehylonierone o The implieehfilone of hhie ebony one then the ehylo

nieron protein m y he inporten‘

h for the eheorption of fee in the inte stine eon

m e one of the eonroe e of the high-onenem y M poproheflne in ple ene m y be the

inhe e‘

hine

e , when the inj ee’eflon of ehylmnflerone eonteining le

‘

oeled A end. B

peohefine into eeeipien‘h none , e. roped. di eeppeerenoe of

‘ehe ehyloe ierone end the

redioeehiefity oeenre . The A peo

‘hein eehivfity eppe ere inne diefle ly in the high »

denei‘oy lipoproeein freehion infli eeefing equi lihrehfion oheree e the B protein

ee‘hiviny doe s not eppeen in the plenum inn

/oil often the initial rapid (H eep

peerenoe of the ehylonierone .”Ernie eeeeee

'ee m e {she B protein fie noneholiz eo

alone with the ehylonieron fee , m fie no eeihle then the n protein is also

renewed. with the ehylonieeone

SiM fi oenee no Hem Re ee eroh : Ahnom e l nehnholi en of lipoproteine

in e eeooiened with new eirenlnhory weee ee e o There he as treee ndoue void

in on? knowledge of the ne‘hn

’

oGM m l of‘

hhe ee compounds ehieh om en be filled

if we e re to m de retnnfilthe ehnom lihie e nhioh e mee

Prwoeefi Confee of Pnoge ee : L The finding; thee the proteine of the

ehyloe ierone end the high denei‘hy lipoproteine oen be eolnbilinefl in eqneoue

Ser . No . NHI» 1 23

PES n NIH

Individual Proj eat Eggpart

B : flamers , Awards , and Publica'

bions o

M, Rodbell, Prowim and Lipid Cm osim on of Human Chyle mH am Lipoyroteim , Sa m m y m , 701 .

m the Natmee of Chylm icron ProteinsJ. Biol . Chem o : in m em o

Serie i No n NHI~ 1 25

l o whee-eta }; of Cellular Phyeielogyend. heteheliem

2

3o Bethe ede , Mexylene

m e m eIndiviheel Peej eet Repert

Phe j eet‘M tle : Sthei e e eh the Spe eie e Diffeeeh ee e end. Stm etumee

Principe]. Inveettgetee e : C oh o hhfc'

fihmeeh,9 State E qG o A qvi et ,

©thee Inve etigetere : Jeehfite Geehe ‘31“Beehnieel o

m e Yeere (Cehehdex yeee

m e t : se e

3

Ohgeetwe : he ieelete ewe. fietem hne the etm ethee er? xiheneelee ee e

free e veriety ea? heeleg ee’

t ee‘

heee e o”hheee ethhie e heve heen m ee etehen

einee it eeeme e reeeeeehhe weehihg hwetheezte that the heme general. etm e==

there m y eeeee m the e etive eeeteee efi’

ell ef the etheeh elee eee, but thee

verfietiene in etm etheel detem m y em ht in theee pert e e? the etm eture e

whieh ere net direetly eeeeeehe e with eetehyefie a

ghee}; penereee h e e been ezeemihee fer ite

rihenhelee ee eehteht end few di etih e t peetefile eezhpeeehte heve been ieeleted

from extreete eff thi e tieeh e by eelhm e M emetegeephy o All few eeem he he

eleeelty reheted e h megahee m leem ee ee ight we em . greases a

“Ehe etm ethe e

of the hem-

fine eheyme em . theee eheegee in m ine eete eeeeeeee heee been fem e o

81mm e theiee ere new in peegf’eee em eiheehelee ee free perh eeeeeee e w e.

free epiheeh leeeee o

hie em the hie em fueettee eff peet e fih meleehleeg geettehlerhy theee having

eh eymetie prepertiee o {the w e}; in gemereh the ef e, m erely hem e em . theeretieel

exert with ehvieue heeetnge eh tieeh e h e teheliem 51 1m genem l 0

en ef m ine h eiles in the e we-eel

rfihe eh elee ee e the enz

yme h em e prehe hly he ieeletee freh e. fee ethee hielegm

heel eeeeee e te extend thi e"eeth hee demehm etee

"eppeee eh he the me eheni ee ef

Ye e X

Serial No .NHI“ 1 23

PBS 0 NIH

Individual Pt o j ec t ReportCalender less 1 9 58

Port A con't .

Pro jec t Description con't o

The blosynthesls of cysts thlonlne from. hono set tne and

cysteine hes never been sheen in c ello tt ee prepst stlons , not have

we so for teen shle to show it ln. hent o sgot s . we feel that the

mechanism. o t this reac tion say he mot e conplen then a simple

elimination o f se tos o one the t it one he conveniently studied

toge ther wi th threonlne hlosynthesls ° tot enenp le o phosphohomo o

set lne csn he tested es s posslhle lntetneels te in cyststhlonine

blosynthesls .

me thods one maj os needless

A t ellehle , eccnt ete esssy tot tht eonlne hes been developed Dwhich require s 5 minutes es cement ed filth 3 on s hos t s for e ssays

previously evsllshle .

One gees . of not e ®o

ohosphohomo set lne he s been i solated from

an ensyms tlc lncnhs ttnn of A t? one honoset lne . The methods of

lsols ttnn and o t pnt lftcstlon of yeset homo set lne hlnsse were

modifications of those ot he tsnnhe ens Shlnnt s .

Of neny nnsnee e otnl ottenpts to presse s na

phosphohomo set ine

chemicallyo the mo st st oni sing tlll son' he s heen nle N e cessa

lsc tone Ne cethoo

bensyoxyhonoset lnehensyloni é e o Snell ess ente of phosphohonoset ine

have been ohtstnes hy tt estnent of the lstte e

slth oolyphosphottc

sold fo llowed by sole hyssolyets o lt ls plenned to tty not e gentle

pho sphotyle tlng seents , such e s te tt e a

po ni ttophenylpyt ophosnh ste .

The t esnlts of wetsnshe concetnlng the pe the sy - tonhhonoset ine

to tht eonlne in yeast have been confirmed , The ensyme convet tlng

pho sphohomo set lne to tht eonlne has been ldentltled ln. hent ospors

wild typ 5297 9 end he s been ent itle s told in e 3° step pt eceednt e o

This enzyme sc tlvlty is onse tec tshle in entt se ts s t notent 35423 ,onset require s exogenons tht eonlne tot gt onth ° Cnt lonsly 9 i t

he s so for been impossible to shos '

the presence s t hono serlne kisses

to Neono spems .

Pyt leonel phosphate hes been shown to he a s elector fot the

enz yme converting phosphohomose t tne to thseonlne .

The fle et studies ot the ne chsnlse . ot the le tter reac tion

have been carried out hy lncnhe tlng pet tte lly out fitted ensyne wi th

substrate and cotsc tot ln Tt ltlnnp leheled e s ter: ° ll tht eonine were

formed through on ole ftnlc lntesmefile te snch so vinylsglyclne o i t

2°

'Serial No .NHI” 1 28

Individual Pro jec t Roport

omit acid to Amflmml Ti ssue s .

Mttaboflism. of Pt opiomfic A t ifl im. Amimal tflsooao .

Ki n Pt tpionyfl Goomo

ymt A Cottoxyflatflom oyotomo

Flowing H o CaGQEQ°MQm@@8a3 amd Soweto Gt hoagJoChom t , 229 9 Qtfl

'

itfififlfi o

‘

Mt ttboitsm. ot tt opfiot it Peta fin Amimmt ti soue t o

I ,

martin

Biol .

Serial No ,NHL- 1 29

Individual Projec t Reportcalendar Yaaa 1 9 53

Me thods

on at to“; ot thogpnoapnato and W e

ayatan, so no phoaphatc cateri o

though a m ic of this

yat been defi nitelyto noon emanat e to anggeat it may inaction in

M o an cm contfly ocean to to tho caaav a m to a cytoe

a, it o ut to the th at one t apot taa tw o a

m pctananta wi th on at tainaa totncnttng otgantan in gt oaingtnt ca natng octnyla taoatannew notatna tantalca that acetate

tncafiaca i n train otgantan tw o the natnyfl m anna of bataioc o

9 in cann inanta with Q‘Mog ant com octatna them was ine

tfion of? ioo tonc into the acetate s contfltafing tna ot caanac

otganflm of an action onao catfoon Etagnant natatou an o

WWQE’

9 tooanaa of alttticnfitfiaa encounten’ca

'

l tn at oning tan organian

a finance. acatc not to otanat tng octane acted canto at cattacta9

cation was attrac ted ins tant! to tfaa atnoy of tho confi ne organian o

A ttcnata o ut to noon to trac tionatc ant pat-many notify tac

:t act Eton tna cfnoMna oagantan no no to be onto to atnoy not e

infinity m) cotactot nonnatonantag £22} antennaaXataa and taac tion

annotate9 {3} nnoaptrata oatotwflcatton o tat ttcnlat attantton ou t

focnaaa on too natnta at the cytocnt onc at a tote tot fita as a

fantat ta contfim cao

( 2) Enantnau y attannta m y. to mania to obtain active

apatatfiona tt on the notation otganton ano to atnafly taxis

montatfion in aatafiao

t E incRadon

Serial No ,PH I- 130

Laboratory of Cellular PhysiologySection on Enz ymes

BeEbesda, maryland

was 0 NIH

Individual Pmoj ec t Report

Calendar Year 39 58

Paxt A

Pro jec t Title s in aexab microazgani smo

Pringigal Inveamigaa@z s

Other fimv@s&£ gatt r

CoopemmtimgUnfifis

mam years

Tb taig 3 0 3 Patiamt Days waneProfessflawalz 1 ,

0&h@r

Pro ject Deggxigmi@mg Sandy she machamfismk of isapxemoi é n gadation o

Esolafii®m @f a mictw©ggamiafiz fi%fich perfoxms the oxydation

of cifiranellofio since fihfls is an fisapz ewwid of smallms leculam aeflghfi fl& represemts am fideafi madefl ta gaudy the basicm@chanflam ofi é egmadafibmo gtufliee wflah dried cells

have sh@am fihat in the pxeaemse fis@v@3egfic aefld amfi cit Qellol

1 abele§ ca 1 9 ihfi®xpofiaae é fimfi® a@e@afie and fia hydz @xym@t&ylglutar1 c

acfid o in fled cenlg ia afisg hag been smmfim fih afi Eabel®d ficezate

is imaompaz afied imao amino aai pr@teflm @md am. ye 1 low'

$esin which

mfigbt be a tefipeme o 1 3®fiatfimn I mwz ififi e flfifiem pgwcadmz e e fox

the sa preducts h&ve . be&m masked aut o

A s a m®gula ©f studies ia enz yms pxepaz atfloms it i s proposed

thaa the ciar@m@ 1 1 @fi fi scuz s as 3h@ww in aha eaclo sed

schmme g Thea@ sh@@m flefiifiicafiy t® eccmz are fimfiicated bythe saflfid arr@wa whefeas bmwkem &EE@W® $¢eps mot yat

Qsmabliahed o

Furthar studies @m degradafiiwm @fi citz anelfial o Evidence s fo :

the convaxz fsn afi cifiramellflc o acfld ta Ba mafihylcxatomyfi=CO A which i s

no t gr é wed yet o Studiaa eath isopramaides of highefi mglecular weight

lika Eamon amd aqualene o Chemical charac teriaaafiom. @ f the yallaw

re sin o Study of tha symtheafis maehawflam of mhfis cwmgoumd o

Pfi f t B ime lufled fie x

Serial No oNHI Q I31

Sec tion on Enz yme s

Be the sda , MMryland

Individual Pro j ecm ReportCalendar Year 1 9 58

Port A c

Proj ec t Tielo A . sanay or the Me tabolism Throe G Carbon

Com mands m the Anaerobic Microorgani am

Principal Investigator : Howard Goldffimo

Other Envoorflgomor

Cooperorfiogwmirs

Man. Yoars (coioodor year 1 9 58?

m en L 3

Profe ssionals 3

3

E0 obtain an onderoromofiog or rho ohemfloofl reactions involved

in tho commeroioo . o § oiaoflme to a mirroro of aoorio and propionic

ocios o Sfiooo those roaorioo s appear to provide rho orgooi sm wi th

energy fior the ofiooyomherio prooooooo fieodfimg ro growfiho i t is

hoped fibre mafia ormoy vi ii loao go on omé eroroofiing of rho energetic s

of rhis orgomiom o

Soroims of this organi sms ohfioh carry omr rheofiorememrioood

roactioos s were isoioro é by the prooooo of enriohmomr ouiture o

A . mo&hod for obtaining ocrfive o oello firoo s crrraors of rhooe organisms

was found goo applied o ioflriol orodfios roooofioo rhor pyruvic acid

was nor am intermooflaro fin rho oom moroioo of glowing to ace tic acid

ia rhooe errraoro o Hooerera pyrorflc ooifi fio metaboliz ed by obe se

orrrocro on@ a maj or marooolfito has boom isolated fin small quontitios o

This compouofi appeoro to be a dioarboxyfiio o cromfloo acid but i s no t

glutoofio or aspartio ocfid o

fihe metaboiiro ofi pyrovio road fialfi ho isolated in qooori tie s

largo ooough forposirivo ié onriffioarioo o rho rolo of rhfio compound

in the ovoro ofii morabofifioo . wiflfl ho studiooo rho cooweroiom of

alanine to acorfi o and propfiooflo ooflé a rail ho erudfied fin rho Eight

of this moo ' imfioroatfion o

Parr B inoiudod

Serial No ,NH1 ” 1 32

PBS NIH

Individual Proj ee t ReportCalendar Year 1 9 58

methods end major Findings con°t

Changes in the settstty of tat toos enz yme s have been observed,and experiment s one tn. pto@t ees to tty end nonet stnnd the mechani sn( s )underlying these ehsnges o the hehsntot ot the tt ee sntno acid pool ,

(which is about ifihx ot the hey e efigh t ot the o et1 ) 9 one the ht enkdonn

and the synthests ot protein at dittet ent etoges of net elopnye t ant

being tnvesttgs teso the peo tetne one seine needs of the eee eeee are

labeled by at onementhcn. on. e methionine e t ennit tng notent ot E , coli

which hon i tself been gtoeflng on $35 e ne thtontne o The distribution

of the lshel is then toM oe ed in nat ions oheni csll Em o tions of the

slime mole st the snnehee o shoe end tente steeno It ne e tonne that

the specific activi ty ot the methionine to . the sh ine acid pool

(hosed on to ten ninhynt fin} one the nt otetno (he sed on hint et ) stays

constant t .e ° 9 me thionine is s tynfieefi seine notes forming a constant

frac tion of the free enino needs end the nenne needs tn. pto teino s t

nil stages ° It is hnonn thst ntotsen in the nttinete source of

the cethohydt ste ( eelfinfio se ) totnee es en ens pendent of different a

tattoo . Folioeeng t enets 9 total nflnhyet tn8 end hint et vetoes it

ens seen thst 9 e s flfltte t entfis tion pt oeeene o tient the snfino setd

pool finite one t eeeine st 1 13 its tntttet M ilne 9 then sfieoholo solnble

pro teins nest eese 9 one tensity n tene tton of the hnflh of the pto tein

( nle ohofi tnsolnhle p Thne petetpfitshle ) st ops heteeen the slog and

fruit stsges ° By following the eefiense of entno eefins genet slly and

methionine pet tionle t lya it ens seen thet the gt eetest proteo lytic

ac tivity oecnt ed ettet the ft ee enfino et td pooh had st opped end at

the time of the height ot nat ions oneynntte ne tivi ties o In V iew

of the dots in the Etteee tnte eoneetnfing t efistionshtns of pool levels

and the ht eehdoen ( end resenthenteb ot endogenons nt oteine o i t is

conceivehle thet the oeet ense in t he enino nook of the shine molds

ht onght shout by stet vntion in some any fini tistes eneoeenous pro tein

hreohdoen ,

Pt efitnine ty nepet finents shoe that e t th e tine of menth ol

pro teolyeis one enzgne se tivtty thet e i s s menthol notnhe of et c

teenslly supplied methionine tnto pto tefin o Kt enpent s 0 thet etote 9

that "tntnove t

w ts enhenoed st this pet toe of diffet enttstton .

serial N0 0 “M a l-32

PHS ° NIH

individual o j ec t Raport

and M, L, fimdez mm o Realm (cage

B o fiv 23 9 370 6119 53)

B o E a m u ms, E’M fic Acid m am m Fox

-ma am M an on ,

Biochemistcty

B e E a wmghftg Effeett «35 Sta mm m m a Aggm gaimgm in the

Sum Mow Dic twsm fiim M sw mm o Em a, 19m m ( 1 9 58 )

B , E . m ime am?! 34 0 E ” Andes-m m m a

gma pan am a dut imgm gfem mmau m in {the su m Mem o mm mm E mma

m pkfims £958 0 (m press) ,

Serial N0 0N31 - 1 33

Laboratory of Cellular Physio logySection on Enz ymes

Bethesda, Mh ryland

lndivddnal Pro j ec t ReportCalendar Year l9Sd

Part A

Proj ec t ti tle Synthesi s , laolatton and Characteri z ation of

Componnde at Biological Inter-eat o

Principal Investigator h e todd hdlea

Other lnaeatlgator

Cooperating Uni ts

Man Years

Pro j ec t De scription

The pnrpoae or the reaearch to to aynthealee compounds for

use as enhatratea tn enaymattc atndtea and to attempt the isolation

and charac teriz atlon or hloloatcal lnternedtateao

Methods and Maj or Ftndlnga

The atrnc tnre at a not r hacterial degradation prodnc t 9 isolated

by t o Z . Smyrnlo tla and h o h o Stadtnan8 one shown. hy degradatlve

reac tions to he lo rlhltyle h plo ddme thylo loz ohgho tetrahydro

o z b é o

ddhetoqulnoaaltne ° The men'componnd haa heen ayntheataed chemically

by too methods o A number of poaafihle netahollc tntermediate e in

thi s degradatlre pathway have alao heen ayntheataed o

Preliminary eaperfime nte on the atrnc tnrea o f three o ther

bac terial degradation produc ts of riho tlandn have been carried

out o ho t i t i s too early to draa' detlntte atrmc tnral conclusions °

One o f the compounds appeara also to poaaeat the dihetoqntnoxallne

chromophore and to have undergone ertenalre modification of the

ribi tyl side chain °

The infrared spec tra of D20 aolntlona or the nocleoaldea

and nocleotldea studded earlier have heen pat on a quanti tativebasi s ° The integrated intensi tie s o r the peahe tn the carbonyl

region have been de termined by hamsay’a method ° The numerical

value s are independent of concentration o

Serial

PMS w NEH

individual Pt o j ee t Report

Calender Yeee E9 S8

Peblieatioe e

Ee feeeee Spec tre ewe Te e toee t ic Stree twt e o f Ne e leo eide e

and Nuc leo tide s fie DZ Q go fie tie e o EI Bioehie o Biephys o

Enft ered Spec te e and Tee teme t ie Struc ture in E2@ ge lution

o t Polyeeeeylic end Pe iyut ieylie A c ide o Chem o end

ied o 59 1

Qe ee ti te tive intree ee Seee tre in $20 o f Some fiec leo eiee e a

Ne e leo ticfie e 9 and Pe lye e e ieo tide e A Nee ' fieaaee e of

Po iyeee ieo tiee Xnteeae tiee c Bieehie o Biophye o A c te s

in pee se o

e a iso lation end Qtree te t e Pee e f e t B oeo eie e teytc ewceeboxy

b

greyeoe e as a Eec te t iefl Qe gt edatien P t edee t o f Ribo flae ie u

and E n R , staete ee o

Bac terial eege eeatioe Proeoe ta o f Rieo tlaetn E1 1 0 i so latioe 9

Ste ec tnt e Be tee e ine tioe p eee Bio logie e l Ttanefot ne tioe s

of

qnieoxafiie e o 3 ° Am e Chem o Soe o g in pre ss o wi th P 0 2 0 Seytnie ti e

Serial we , NHI - lBS

Laboratory of Cellular Physio logySec tion on Enz yme s

Bethe sda o

'

Maryland

PBS NIH

Imeividuet Pro jec t ReportCalender Yeet 1 9 58

Pro j ec t Ti tle A : Intermediety me tetc ttse of emte e acifls m.

particular emehest st em the te t eectiemc

fieve lwimg evet ell t ede é tflve deemime tie n é o

Aneet ebtc e e tebe tiem. ot e etkyio

gt oup ficmox

c empommds emd N s eeimmxceepe emde o ( tee t epcrt

it em Et a Hugh Heywetd)

Principal Knee sttgetct : Eet ecee C o steetmee

Othet Investigator

CooperatingUni ts

the years

To tal

Profe ssional: 1

Ozhet £ 0 3

Pt e j ect De act iptien

Obj ec tives Tc ot tatm finfe tme tiem cemcetmimg the mature at

the enetgyc t tch intermediates termed ehem vet flcus amine acids

undergo t educttve deemtmettem te tome the cett e spemeflmg fatty acids

and aememie o Ai ee te study the electt ee tt emstet t eec tiems ( anaemcbicosteo a t edee tiem reac tions) int e fivee it these pt e

c e eeee o

A meeedteme o eepeedee t phe ettetese that was etecet e t ed in

pat tielly put itfled glycine cedectaee tt ec tteme pt epet ed tr ee

studiefi fie acme detail end pet tfially

ewe we e po ctt teted te be ” instead , a tt emstet ase which , in the

presence at thi s umet tut el qmimeee o behaved cmly e e a hydt c laee o

0c the beets ct thi s aseumpttom, stedie e were made on the glyciee

t eeuc teee system t egeceteg the pe eeitfie perticfipe tiee of e quimcne .

This i s cf particular interest for the exfide tiemo t edec tiee reac tion

in que stion ieede to the formaticm at A tt s A c j udged by a number

of criteria such as mat ted eemattttfity to emtteyctn A , inac tivation

Serial No ° NHI~ 1 3§

Publications :

MEtaboliem. o £ Steroidao Chapter in" Cholesterol” Ede by Robert P o Cook Ae edemic Pre ss ,

Inc e 1 9 58 . 9 0

2° A memedfieme m eepeheeh e enz ymic hydee lyefis of pe hitrephenyl

phe ephe te ° Jo hiel. Ghee » ( in pre ss)

The pereie ipe tie h eh a qeiheh e in hhe eheyehe eedee tion

( he eee ee } o

Eeeeee e ieee heeeheeh h ehhefliee phe epheee he o f yeest ,

A eeh edieh e depeeeeee ee e

ymhe hyeee lyeie h f pc h iteephehyl

ehe ephehe o Emeo Gehgxeee fez hfleeheefle 9 hieh o 1 9 58 ,

6 ° Xe pheeeeehfieh efiehJo hez eeee( I) The feeeemhehieh efi o eeih e eeleee te hy cle emxidium

emfieeeeleeieh h h (h©e o eh o )

( 2) The feememhahfieh e h 7a eefih e hh eyeehe hy mhe eEeidium

N0 0“HIG H ?

done in conjunction with Dr eJohn Fahey of the Cancer Ins tituta>He also has characteriz es. the gamma globulins by di a ethyl-andno

celluloe colunm chromatography, and our tests of the eluted

fractions show that the chyroifi tis ears have "normal " gm

globulins whi le the antibodies against nucleopm teins in lupus

ears are in the elution fracti ons which usually contain macro=

Bo animal inmmnizafi on studiess Witebsky and Rose suc c

ceeded in producing sutoantibodie s to thyroglobulin extracts and

correspondingthyroid lesions c We have failed to produce detectable

antibodies to rabbit nuclcopm tein extracts when rabbits were

irnrm niced with rabbit nuch oprotein extracts o But when humannucleopm tein extracts were us ed to inmmnise rabbits, antibodieswere produced not only to human rmcleoprotein extracts but also to

rabbit nucleopvotain extracts, Theso rabbits now have circulating

gam e globulim which react with their own nucleopx'oteins a 1 0 9 0

an experimental analogue oft

the L. E0 factor o So far no evidence

of tis sue damage has been foundo

Significance to Heart Research : These caperiments indicate that

profi tion in sam e 0? em bodies to foreign nucleopnotsins

(c ogo bacterial) can produce antibodies which also react with

native nucleopm tcdncg i ce “ the experimental analogue of the L0E

0

factoro If confixmeci, this data will provide an em erimental

basis for the hypothesis that lupus erythenatosue is a disease of

auto e antibodies , and um vide an explanation for the production of

such autoa ntibodies 0 Although these experiments were intended

origim lly to test t he hypothesis that auto mantibodies are re sponsi a

ble for human glomerulonepM itis, so far no evidence of such

antibodies has been found, although the search continueso

Gam es of one Pro ject s In conjunction with Dr, Fahey (mi )s u

'

of characteristics of the gamma globuline

which react with thyroglobulin and nucleOproteins will be carried

out, Im unisafl on with nucleops'oteins will be continued to see

if antibodies to nucleoprotan will produce tissue damage in

experimental animals similar to that found in clinical lupus

erythematosus o Fur ther attm ts will be made to produce auto :

antibodie s to rat kidney in light of the preliminary successful

reports from Dr " Hem ann at Western Reserve Universityo

Part B included a Eg o

NHL-1 37

Honors ,

Publicati ons othe r than abstracts from this pro jects

Goodman, H oCo and Baxter, J o P a The Search for Anti »

Kidney Antibodies in Patient-s o P mceedings of the Ninth

Conference on the Nephrotic Syndrome , p , 61 1 9580

Hom rs and Awards relating to this project :

PHSo NIH

Indivi dual Pro j ect ReportCalendar Year 1 958

Serial No nNM ° D 8

canoe to Research Invest igation of hypo

0 heart research,increase understanfiing of mechanisms

In addition a minor degree of loss of

the GI tract has been demonstrated in

isn‘t with manifest heart disease and a. marked degree

in anothero It is possible that this proce ss is of

(em eielly constrictive pericarditisfi

Serial No e “HI- 139

Physiologyr and Metabolis m20 Section on Metabolism

30 Bethesda

Individual Pro j ect ReportCalendar Year 1 958

PART A c

Proj ect Titles Studies of Inhibitors of Cholesterol Biosynthesis o

(Started 1 955 a» not completed)Principal Investigator: Dani el Steinberg

ones Investigators g no so Fredricksons J o Avigan, E 0 130 Feigelsono

Hugh W omen, Technician

CooperatingUnite s None

Men Years (calendar year 1 958) 3 Patient Days (calendar year 1 958)

ProfessionalOthers 0 25

Project Descriptions

ng com m cholesterol level by m y of inhibition of

cholesterol biosynthesiso

fed synthetic diets containing the

sacrificed at various time intervals o

Serum and fl e ece cholesW ol concentrations are measuredo is

e sense of the rate of cholesterol synthesis the uptake of

G =labeled acetate or ne celonste is determined just prior tosscrifi ce o Effects on sdrensl steroid production are measured

by assay of adrenal vein blood obtained by direct canmflntiono .

Clinical shadiee entail measurm ent of serum lipid

concentrations and also periodic evaluation of liver M otion

and kidney function as s ell as hematologic findingso

an action on acetate activation end are compatible with the

negative clinical results with this compound and with Baphenylc

O

(2) The previously reported effects of Al‘

ccholestenone on

adrenal function have been confirm é em extendedo Simultaneousadministration of adrenal steroids prevents the adrenal

Serial NO 0 NHIQ B 9

Individual Pro ject ReportCalendar Year 1 958

9 , Awards, arfl Publicati ons

na other than abstracts from this pro j ects

R etin a-

son, ms”, Peterson, and Ste rg, Do

on of adrem cortleal steroid secretion by A wholestenone ,

e 1 27: musics, i958o

Masters, R, and Std nberg, Do Studies on the mechanism of

of a hao phenylbutm te o Brioczhimo at BiOphyB o Acta_21 :

79 1 95 o

Steinberg, D o , Fredrickson, and Avigan,Jo Effects

stenone in animals and in man, Proc o Sm: 0 ficpt o Biol ,

21 3 7614 0 790 9 1 9580

Awards relating to this proj ect s

Serial No eNHI-MO


Part B: Honors, Awards, and Publications

Publications other than abstracts from this pro ject :

Fredrickson, mafia Ono9 K9 ,

An improved technique forthe measureme nt of Garbo dioxide in emb ed air, using the

liquid scintillation (Bonnier o Jo Lab o Olin, Medo 51 : 1 147, 1 9589

Fredrickson, D 0 5 0 , McGon ester, D oLo ,Havel, EQJO , and

Ono , K0‘lhe early steps in transport and metabolimn of exogenous

triglyceride and cholesterolID Chemistry of Lipids as Related to

A therosclerosis, I. no Page, Editor s Springfield, C .C o

‘

l‘

homes 31 9589 p a 205°

Fredriokson, D°S . , Mccollester, D 0 1 .” and Ono,Ko ,

The Role

of Unesterified Fatty Acid Transport in Chylomicron Metabolism,Jo 01 1 m, Invest o 21 : 1 333, 1 9589

Fredriokfin and Gordon, Jro The Metabolism of

c i ebeled Uneste rified Fatty A cids in NormalHuman Subjects, J Clin, Invest “ 21 3 1 5011 9 1 9589

Fredrickson, end Gordon, R.s, ,Jr o , mausport, of FattyA cids s Physmlo RfiV o 2235854 30 3 1 9580

Honors and Awarde e

Serial NO 0NHIQ MJL

metabolism and serum protein levels 0 Since through such relati on=

play a part in determining serum cholesterol levels

s ell as in vitae effects of epim phrine on adipose tissue are

sted, Relation of lipidenia to the calorigenic effect of

(2) Binding studie s in isolated system are continued withhigh density (a) lipopm teins m ien appear to bind UFA more

tightly than the low density lipoproteins o A s the affinity of

UFA to lipoprotcins seems to increas e wi th the chain length, the

UFA betseen

Serial N0 0NHIQ M

1 ° LaboramPhysiolog and Metabolism

20 Section on Metabolism

3° Bethesda

Pee-en'

s


Part A u

Project Title s Cross-linking of Proteins M th Fom sldehyde o

(Pro ject started September , 1 957 completed June 1 , 1 958)

Principal Investigators Elemer Mihslyi

Other Inves‘

d gatorss Me I n Knoller

CooperatingUnits 2 None

Man Years (calender year Patient Days (calendar year 1 958)Tom : 1 9 0

Professional : 0 0 5Other: 0 0 5

Pro ject Descriptions

ohge ctives z To study the mechanism of reaction of formaldehyde withpro e with special referem e to gelation phenomenao

Kinetic investigation of viscosity and Opacitya]. analysis of the fom eldehyde treated materializ e

conditions are widely different with various proteins a Although

the um ber of proteins studied so far is small, tee cleara cut

classes can he distinguiske ds (l) fibrous proteins (fibrinogenand myosin were studied) 9 which form a gel at relatively low protein

(L G?) and formaldehyde concentration (l%) with a pH optim nn on

the elm side of the iseeleotric point (2) globularproteins (bovine senm 5.1 t and e g albumin were studied) ,which gel only at higher motsie (25 and high formaldehyde cone

centration 3 in s re rros pH sons around the isoelectric point s

The mechanism of gel formation was studied in more detail with

conditions were compared with the native proteino fi lers was no

differences as compared with the native protein, in the number of

tyrosine or tryptophsne groups whereas e marked decrease

« soa red in the number of lysine «é‘

« emino m ops in those samples 3and only those , which formed a gel o Using Gm formaldehyde it was

Serial N0 0“M a l

1 0 Laboratory of CellularPhysiolog and Metabolism


3° Bethesda

PHSsNII-I

Calendar Year

Part A c

Project Title ; The Hyperlipemia of Experimental Nephm sie z SomeInvestigations into Pathogenesis o (Project started

Aug<1 l, 1 958 not completedo )

Principal Investigator: J 9 0 0 A llen

Other Investigators : J. H o Baxter, H , C o Goodman

CooperatingUnites Hone

Man Years ( calendar year 1 958) Patient Days (calendar year 1 958)Totals

Professional :Other : 0


Obgectivesa The hm riipsaia of human nephrosis is reversed by the

a ration of elbmmin as sheen in this laboratoryo It washence decided to study the effects of altering various conditions ,such as diet, serum osm tic pressure and blood coagulability, on

ments may help elucidate the m thogm esis of re phrotic hyperlipeuda9as well as explain the response in patients as mentioned abovc e

Methods a. - s _.a hephrosis is indm ed in rats by the inj ection of

an are;t serum in a manner previously describedfrom this laboratoryo Methods for determining serum total lipids 9total cholesterol am phospholipids have been adapted for microe

quantitative detm inetiom in these animals o Using these methodsserum lipids may be quantitated before and after the applicationof various experimental conditions o

Meier Findigss foe adninistration of dextran in hyperlipemic

nep c rats causes return of all sem m lipid values to normal

levelso Short term adsdni strefi on of a carbohydrate polypeptidediet causes reduction of the high lipid values, which return to

more abm rmel levels on a ins o fat balanced solid chos e

Sharia], N0 0MM ” M

1 ° Laboratzry of CellularPhysiology and Metabolism

20 Section on Metabolim

3° Bethesda

Individual Pro je ct mportCalendar Year

as Studies on Soluble Protectswe Substance inFbrperimental “whritis o

(Project started Feptemberg, 1 956 e not finished)

restigatore James Ho Baxter

gators: Patricia M fy, Howard Goodman, James Allen

fnits : Mens a

slender year 1 958) Patient Days (calendar year 1 958)

L1 : 0 0 1 none 0

ptions

To study and purify the soluble substance derived

m t tissue , which will neutralize

oluble protective substance is present inon prepared by digestion of rat kidney or

on obtained by digestion of acetone

tit s and d omains less inert m terial

d by digestion of kidney hom genate o

a or preparative

insctiVation of the

igen being studied may

9 end nephrosis o

Serial N0 0 NHI'D MS

l o Laboratory of fiellfiarPhysiology and Metabolism

2° Section on Metabolism

3 Bethesda

Indivi dual Project ReportCalendar Year 1 958

e ject Title : Proteolytic Fragrant-su ch of the Myosin Molecule(Project started June 1 , 1 958 completed Oct o 1 5 1 953)

incipal Investigators M ower m ay—1

her Investigators: William F 0 Harrington, N o I. Knoller

operatingUnite s Section of Cellular Physiolog'

n Years (calendar year 1 958) Patient Days (calendar year 1 9 58)1 10 1781 3 1 0 5Professional : 1 00

Other :

e ject Descri ptions

Objectives z Myosin is a very large molecule , much too large to be

attacked by structural investigations in its intact form? It was

thought that the proteolytic fragmentation of the myosin, since

it yields sell defined framents , say be used advantageously in

structural investigations , especially to establish connectionsbetween the different functions and certain localised. areas of

the molecule and, on the other bani , the proteolyti c processitself may shed some light on the structure of the molecule o

Methods lo 3 Vis cositys optical rotet ion9 sedimentation? pH e stat

no

ation of the myosin molecule was followedsome tine the number of peptide

bonds split was also determined by physicoohenical methods and

also confirmed by ohm ical analysis-

o It appears that some of the

pmtide bonds are split at a rate about ten times faster than that

of the rem inder s

"This fast reaction accounts for the formation

of the large fragments o Sons optical evidence suggests that parts

of the myosin molecule are formed of a tightly coiled chain,whereas oth ers are much looser and randomly foldedo Apparently

the enzyme attacks with w eb greater ease the random portions and

thus liberates the tightly coiled fragment s, giving the erroneous

impression of liberation of preexistent subunits o This reaction

Serial No e“HI-Me

lo Labora ry o e ar

Physiology and Metabolism2° Section on Metabolism3° Bethesda

mSe NIH

Individual Project ReportCalendar Year 1 958

Part A o

Pro ject Title : Development of a Method for CountingWater Soluble

Compound s in the Liquid Scintillati on Counter o

(Started Aprils 1 958 e not wmpleted)Principal Investigatore Daniel Steinberg

Other Investigator Ray Pittman

CooperatingUnits

Man Years (calendar year 1 958) Patient Days (calendar year 1 958)Total : 0 2

Professional : 0 1

Other: 0 1


Objectives: To make it possible for the biochemist to obtain radio=

assay on polar compound s directly without the need of first

converting them to an orgm ic soluble form Presently available

methods for dealingwith eater soluble compounds are either limited

to sm ll quantities or inm lve time consm ing conversion procedures o

Methods E lo eds The basic principle is the use of a teo o phase

system a sofi d phase consisting of finely divided fluorescent

material and a liquid phase containing the compound to be assayedo

The earlier stufi es utili sed a plastic with diphenylstilbem in

it (pilot B) ; later it was ifo that crystals of anthracene

and of diphenyloxaaole aere also suitable for the solid phase ;

Ma

Qor Finding By reducing the ratio of fluid volume to solid surface

halfi

been possible to shorten the mean path for the beta particles

of 0 prior to their collision with the fluorescent material o

With the plastic scintillatee m adam efficiency was obtained using

tightly packed filaments and adding the aqueous or alcoholic solu

tion into the fire intersticeao Efficiencies up to 29%wereobtainedo Using a large number of short segments or beads erric a

iencies of 1 0 to l% were obtainedo The sample can be recovered

unchangedg which is a dis tinct advantage in some studies :


Awards, and Publications

Publicati ons other than abstracts from this pro ject

Steinberg, Do Radioassay of in Aqueous solutions usinga liquid scintillation spectrometero Natm '

e

Steinberg, Do Radioassay of and tritium in aqueous

soluti ons in the liquid scinti llation spectrometer o In Proceedings

of the Symposium on Tritium in Tracer Applications o New York City,Oct . 31 , 1 9580 In Press o

Honors and Awards relating to this pro jects

None 0

e 2 a Serial No e“ P M?

lipid c ontent , but produces minimal chyle , suggestive of mucosa].

absorption, but reduction in chylomicron synthesis, The effect

of ether anesthesia in reducing chyle production is well known,but has not been studied in detail ,

Significance to Heart Research : This research is intended to define

in more detail the chenfstry of chylomicra, the large lipoprotein

by which fat passes from the intestinal mucosa into the blood

stream, Patients who cannot remove chylomicra from the blood

stream are known to have an increased incidence of atherosclerosis,and atherosclerotic lesions are a major cause of myocardial dis ease d

ave succeeded in outlining the majorn of such a system for fat absorption,

and should be able in subsequent months to succeed in getting at

least some fat absorption under the proposed conditions , The

mechanisms by which anesthesia inhibit s fat absorption may be of

physiologic significance , and using thoracic duct cannulated animals ,we hope to study this in more detail , specifically, whether factors

which are know to be released during ether anesthesia, e ogo

epinephrine stimulate fat absorption, and whether the alteration

of intestinal motility by intravenous serotin can counteract the

effects of ether anesthesia, These studies are to be pursued after

the completion of the initial pro ject , namely the demonstration

of chylomicron protein by the intestinal mucosa cell o

Part B inclined: Mo o

Serial no oNHI-MB

much larger concentrations of

labeled cholesterol than those which can be readily achieved

by feeding or inj ecting labeled cholesterol . Consequently, theprocedure may be of great importance for metabolic studies

particularly when the fate and distribution of circulato

cholesterol has to be followed for prolonged periods o (2 The

the removal of

cholesterol from the cells and it

place by bindi ng with a preexisting li poprotein complexo

e proj ect has been completed and the

ationo An analogous study designed

to incorporate triglycerides lipoproteins is being

plated o

B includede Mo o

Serial N0 0NHI=M9

Calendar Year 1 958

Sig ifi csnce to Beast Research: The in vitso system devised

studying the synthesis of serum lipOproteins o Fran such 9.

of blood lipid levels , Knowledge of such controls is at?

prime interest in understsndm the pathogenesis and possible

prevention of atherosclerosis

of Resesech 1 ) Studies desi to identity

nearly completed . 2) Studies site in progre ss on the

physiological factors effecting lipoprotein synthesis . We

propose also to attempt to work with cell- free systems in

sid e: to study the more basic sem ets of the links.@ of

protein and lipid into lingoprotein°

Part B o Included Yes o

Serial lilo °Mil- 1 50

project

Goodman, E . c. and Easter,Jo H o M accorticotm phjn

2 Serial N0 0 NHl e lfil

PHS -NmIndividual i ect Report

Calendar Year 1 958

corporation of all of these amino acids is inhibited bydinitm phenolo The incorporatigg

fl

of norleucine (present at

a concent tion of ca . 1 9 5 x 1 0 is inhibited roughlyby 3 x 1 0 leucine or methionine but not at all by lo

m um-3M?

In similar studies it has not been possible to demonstrate

incorporation of norvsline , o-metMl serine or 6- 2 thienylalanine o

A non protein (m soluble ) fraction of liver from which free

amino acids have been m o ved sith an ion exchange resin has

been M d in several studies , The incorporat ion of radio

activity frmn labelled amino acids (alanine, phenylalanine 3lysine leucine and the fluom hm lalanines ) into thisfraction by rat liver slices has been observed . Sons ox this

radioact ivity undoubtedly represents acidic degraded;ion

prfl ucts of the labelled m ino acid s But in the case of

lysinea phenylalanine, p -fl uorophenylalanine there is in

corporation od radioactivity into this fraction which is

recoverable as amino acid after acid hydrolysis o The formation

of this"bound amino acid is inhibited by dinitrophenol o

The nature of these radioactive compounds and their role in

metabolism m not been eldcidatedo A batch oi’

this material

mm the liver of a rat which an received a mixture

of labelled amino acids prior to sacrifice m s fractionated

by chrm ntom pw on a som e Several peaks of radim tivitysens foundg overlapping but in most cases not coincidingwith peaks of 260 absorbing material e (It has been foundam most of the nucleosideg mono

»

,9 di e» m d triphosphates

cm be sepam ted m this column. smim ch of the 860

absorbing material in this M iw likely represw ts know

nucleotides) On further W tionation of certain samples

the W absorbing material was separated few the radio

activity and hydrolys is oi these radiom tive comm yielded

in each case several amino acids (most of them not radioactive )

essentially completed except for some studie s in progress

which may indicate what amino acid (a) is replaced when

the fluorOphenylalanim s are incorporated o Em a-intents in

prom ss are deem ed to give a rather couplets survey of

the amino acid containing compounds in this liver preparat ions

Since heretofore the fractions em ined ( chosen e ssentiallyat raadna) have contained a surprisingly uniform group of

amino acids, it will be of intere st and possibly of assistance

in detem ining the structure of these compounds and their

metabolic role to know whether the compounds do in fact

contain only a certain restricted m p of m inc acids or

whether this early im ression is due to inadequate samplingo

Part B o included. Yes

Serial N0 0 NHI= 1 52

lo Laboratory of CellularPhyeiologr and Metabolism


3° Bethesda


Title : Studies on the Mechanism of A ction of Dietary Fats

in Relation to Serum Lipoproteins o

(StartedJuly, 1 957 not cmnpleted )

Principal Investigators : Daniel Steinberg andJoel Avigano

Other Investig tors: hugh Wrm an

CooperatingUnits : hone

m u Years (calendar year 1 958) Patient Days ( calendar year1 o 5

Professional : 0 0 9Other 0 0 6

Project Description

protein and cholesterol metabolism effected by dietary rats

and to explore the mechanisms or action involved o

Methods used in anim l studie s are described

annual reporto Clinical studies have been

carried out using liquid formula diets contan either

coconut oil or an unsaturated vegetable oil contributing 6%or the total caloric intake Patients received h- Clli- labeledchole sterol intravenously, given in the tom of a om nien

with the patient”8 can serum lipoproteim ( see 1 958 annual

report of Dr e Avisan for method ) 0 Serum sm ples are taken

at interm ls for determination of cholesterol level and

cholesterol specific radioactivityo Cm plete fecal

collections are made m d the excretion of radioactivity in

the form of sterol and of bile acids is determined

serum cholesterol levels has been confirmed o

Serial No e5131 4 53

l o Laboratory of CellularPmreioloar ami Metabolism


3o Bethesda

calendar Year 1 958

Part A ,

Project Title : m taboliam of l na Uneeterifi ed Fatty Acids(Started in 1 955 not yet completed )

Principal Invest igator: Robert S o Gordon,Jr ,

CooperatingUnite : mone o mus proj ect is partly in common with

proj ect of Dr.) Donald S e Fredrickeonof this Sectiono Please see his report ,

Man Years (calendar ite m 1 958) Patient Days ( calendar year 1 958)$0t8 1 : l o 25 50Professional

unesterified fatty acids have been covered in the report byDre Donald S , Fredrickeono

In addition to these inveetigatiom g in vitro studies

utiliz ing adipose tieeue of em eimental animals have

demonstrated that this isolated tissue will produce unesterifi ed

fatty acids 0 The role oi vae ioue hom onal and nutritionel

factors in controllm the output of eeterifi ed fatty acids

is being investigated o In addit ion» it has been demonstrated

that heparin will eauee the liberation of lipoprotein lipase

procese of W A releaee ia hem etudiedo

ican ec_Eeart Eeee The etudy of the metabolian of

lipids is felt to be ofw m ee in the ultimate uader

standing of atberoecleroeie o

Part B o included Yes

Serial N0 0 NM ° 1 525

l o laboratory of CellularPbysioloar and Metabolism


3° Bethesda

Individual Prodset Report

Proj ect Title : Relation between Unesterifi ed Fatty Acidm tabolism and Lipoprotein Fom ation and

(Proj ect started: Februaizy, l957-not om pleted

Principal Investigators : Desist Gm m d Daniel Steinberg

Other: Halter Xe nia

Man Years ( calendar year 1 958) Patient Days ( calendar year 1 958 )

Professional

Proj ect .Description

A numbei" of fatty acid analogues were explored

a compound which would successfullycompete with normal fatty acids for bindiw on serum albumin o

With such a compound it is hoped that it will be possibleto block the normal tm spoi

’ t of use sterified fatty acids

transpoxt by new altem atit s mechani sms 3 presumablylipoprotein synthesis and transport o

synthesiz ed, tritiated by the hu sbaoh method9 repurified

by countercurrent distribution and administeeed to rats ,

Major PM : It was shown that this oenpound is not

metaboliaable by the sat at any siwificm t ratea recoveries

being over 9035 sites at houssa 312 vitso studies saith rat

liver slices again showed little or no metabolism

acid o ho mashed ehm ges in sew n lipid levels oeeum ed o

Serial No eNHI~ 1 5&

Calendar Year 1 958

proj ect

GooM an37 D o S o and Steinbergg D o Studies on the metabolism

acid analogue o Jo Biol e Chemo £ 1331 066-7l, Nov a 1 9580

Honors and Awards relating to this pro) est None 0

Part

Serial Ko o NHL- 1 55


(Major Findings continued:) it s search continued for a

laboratory animal that will respond to the feeding of

saturated and unsaturated fats in a manner similar to manoThe mongolian gerbil was found not to o A g imp or squirrel

It was discovered that the feeding oi corn oil to rabbits

previously made athem clerotie increases the rate of

rem ssion of the lesions when caressed to similar animals

fed coconut oil o This is the first time a substance hasbeen forum which increase s the rate of regre ssion of lesions ,

These experiments are being rem ated eith more control

material to incre ase the statistical siwiticance or the

Experiments firm this laboratory in both rat and dog have

shown that fat absorbed in the form of W lw i crons goesdirectly to the tissues Gotham and the Donner m y have

claimed9 without published evidencea that in man they are

deg adm in the blood streams into lipoproteiss of increasingdensityo Em eriments are M ersey to confirm or deny this

coronary heart disease , which is the leading cause of death ,

Serial to oMRI- 1 56

l : laboratory of CellularPhysiologr and Metabolism

2. Section on Metabolism3. Bethesda

Individual Proj ect ReportCalendar Year l958

Proj ect Title : Studies of HyperlipidW c States in Humans(Proj ect started 1 956 not w aisted)

Principal Inve stigator: Donald S o Fredrickson (Studies on

M ocholesterolm ic m ats in collaboration

with Dr : Steinberg ( of : his report for detailed

coverm or proj ect ) :

Other Investigators : Dre hum an Medellester (untilJulyl, 1 958)th e Katsuto Ono9 technician

Cm m ting Units

Man Years (calendar year l958) Patient Days (caleM ar year 1 958)

i'ctal

Professional: 0 0 1 patient hours

Other 0 0 1

Proj ect Description:

Objective s : the observation and study of patients withabnormal concentrations os

’

the various blood lipids with

emphm is on patients uith ao- cslled essential

M erlipidemiaq Data is accumulated bearingespecially upon elassii

'

icationa prom os and insight into

mechani sms of these diseases we. to means of treatment :

A saahly outpatient clinic is maintained at

s re ferred because or siw ieant hyper

lipidM a are seen: Total lipid m slysesa and ram lyglim rotein a n, s rs are ob tainedo Appropriate patients

are selected for continum follow-up m d a number become

available for both out and im patient studies related to

lipid metaboli w m d hypocholesterolenic agents :

available for certain of these studies :

Serial No . NEE

Calendar Year 1 958

Pm B o 30m m , Awards am Publications

Publications other than Abstracts m u ting to this proj ect

Fredricksona D , S o , Petoroong R e E 0 am StombergfiD o ,

Inhibition of odrem cortioal oteroid secretion by A

cholestenone : Scianco, £24 : 7034-705, 1 9580

S einberg, D o , Frodxickoon, D . S o , and Avigan‘,Jo Efi‘

octs of

A whole stenone in animals and in man, Proc o 30 0 0 Expat ,

Biolo and Mad a 21 : 784 1 9580

Fredrickson, D " S o Atherosclem oioa Chapter 8° Metabolic

Disturbances in Gliuhical Medi cine ? G o A 0 Sum , Editor,London, Jq and A 0 Churchill, Ltao ,

Fredricksong 120 S o , Gordon, R e S o ,J'r o , and. Orlofi “9 JoCardiovascular A spects of Metabolic Disease ; Chapter 6? Ibid o

Fredg'icksons no S o Current Attitudea about Athorooclexosis

GP » 93: 1 02- 1 06, 1 9580

Honors and Awards relat ing to this proj ect : mono

z Serial no :NIH - 1 57

M ivifinal Project Rams-t

M ar Year 1 958

b) Infusim s ot hrm cw sod m

m tients sith tm chsnm to the tm l

occur in am ]. centrcls. Um steeii’

ie l fatty acid

D 1 0 019 M W O

bese he am alo

in cm oi“D 1 5 039 - 1 9 063, she the sew hem

elm Ih sflfiitim to this effect , albumin M a m e

m m gheeeith either lactesm t oe elheh W e

is we w the ca ses w tm fi M m o

studies

“

rate or m W e m phrotichy injectionsW e serum.»

Pert h .

Serial'

NooU 3 .

1 0 Laboratory of'

Chemistryof Natural Products

2°

3° Bethesda9 Mary land

PBS 1 1 1

Indih 1deel Prej eet RepertCelehder Year 1 958

ermes1 e A lkeleies

°rfine 1 pal investigator H o 1 0 Lloyd? Ph O D .

athee Ineestigeter Antoinette Velasquez (technical)

eeperetiegUnits

lee Years Patient DeysTetel a75

Prefeseiehel Nehe é

Ohher

”rej ect Deseriptieh

Jerk he the Gewes1 e elheleids eemeeeh 1 ee 9 ermesie1 ee 9 pahemime

thyieytisfiee hes eeeefied theeegh te the pebfiieetiem stage o

31 1 3 week hes wheeetehee eeeeeee hf the peteet hypeteesiee activityred fer exypeeee 1 he o A eeletee eeepeeee9 of the same ether and kind

1 1 1 1 3 9 was prepared 1m m Ne eethwpeeemiee q These substances are

zfien pxeeeets 0 1 eeepfiex eteeeteee g fie the eese ef exypememiee the

cempeemd hes the ehpfirieel fiereele 0231 331 30 Ne ferthee work?1 emet described 1 h keet yeee

“e repeet eed 1 1 the paper9 was

need on the N3seefiee e1 elkeEefieeo

A seeeee see1 es e1 eeeeeeheeg*w1 th only th e e 1 teegee etems 9 was

at eeeee eeee1 1 ee steej o These were 1 eehe te heee the emp1 rieel

la Clgfigga zgfifi The hej ee eeeeeeee 1 1 1 leetem. eee eeeehdery amine

59 eed terh 1 eegmehhyfieee hhseteeet 1 eh o The sfiee ehe1 e 9 cerryiegheelehe eee gehweg hes eeeeeeee stepefise , It hes fehme to be a

:erhee ehe1 h o The th e hfieeegeee eehee 1 e 1 hg e 1 eee hee properties

plieg hheee 0 1 the speetefihe 1 1 hp1 h ) efihefiefies o

D1ree1 1 ee e1’

cheeeht Research“ HW M p 4 m

This eerk h es teemiheted ihJeflyo

3 included

Serial No °

NHI' 1 59

1 ° Laboratory of'

Chemistryof Natural Products

2°

3° Bethesda9 Maryland

PHS NIH

Individual Prej ect ReportCalender Year

Part A ,

Proj ect Title 0 Amine Oxide Reerramgememrs °A

B 6 Organic Bases ofi'

flemem Origin

C o Base Methyietiem Studies q

D 0 Frame Organic Bases°

Principal Ineestfigeeer Mo S b Ffismg Ph .D °

Other investigeeer N , we Johnson (eeehmfieekfiE a 9 ° Lewreeee (teehefieel)

Ceepereeimg Baits

Neme o

Peej eer Deseripfifiem

A O Amiee Gxfiee fieerreegemeeES z A seedy ef the rearrangement

of NgNe dfimeehylrrypeephee exfiee eeder bfielegieel eeedfimfiees was continued a

The earlier chemieefi eerk demeeeereeed thee e reerremgemeem reeeefien

leading to c emeehyleefiee eeeerree readily with ferrie flew ( in a com

erdfieeeiem eemplex) es e eemelyer9 end the eefieeeee wee eeeeieee fer the

existence ef refie eeme e em eeeyee=eeeelyeed bieflegieel dimethylm

eefiee reeeefiemo Three re e xemimeez (e ) ewe effeem ef pretein

deeeteretfieeg (b) rhe added eefeeeer reeeireeemee eed (e ) eemperetivereeceiees with related eeeeeeeeso The fifiree effeee wee smeared ie the

msmel wey by eefiafieg the enzyme prepereefieeg tee eyeeem mes ieeetieegand me reeeefiee ef eey kind wee ebeereeeo This is eeideeee supportingthe eeeyee e eeeefiyeee weeere ef eee reeeefieeg eefieh gees emeeemly with amouse afieee hemegeeeee , re is wee eeeelesfiee evidence 9 heweeer9 since

preterm preeipfimeefiee ewe eeeeeereefiee'

e figee welfl be regereee as remeeiegan active meeel~eeerfieg bee mee e eezymeeie eeeefiyeeo The cefeeeer effects

were steered in the meeefi weyg med it wee feeee thee we added eefecrers

Serial N0 0

NHI' 1 59

Piptadenia snuffoe This hallucinogenic material from Venez uela was

examined for indole bases and it was found to contain bufotenine 9bufotenine oxide 9 and four other indole bases o acacia longigoliao

v This

was found to contain two indole bases° A brus precatorius o= This was found

to contain four tryptophan deriratires in addit ion to nonomethyl tryptoe

phano Other plgnts oe A number of plants reported to contain hallucinogenic

agents were examined° host of the tests for organic bases were negative

and no extended inuestigations were made c

Work on these problems ended in Julyo Further work will be con»

cerned with bringing some of the nearly completed proj ects to a terminal

stage ° The amine snide work will be continued with emphasis on other

aspectsg including the mechanism of the rearrangement reaction° The

enzymatic conuersion of tryptamine is under study in another laboratoryo

Part B included

Serial No e”HI “ 1 60

Chemieelom Further freeeiemetien by chromatography will be

studied in order to ebeeie pure samples ef the glyeelipfiees o Furtheresseyieg ef spiegesiee and feeey acid will he made ween e gas chreme=

tegraphy instrument o

Meeebegismo The bieeyeehesis of the giycelfipides from simple

preceesers will be studied using heme mer we es the seeeee ef the enzyme

syseee o The essay method will utiliz e Cl w iebelfiee geleemese °

Feet 8 iecleded

Serial No o

1 0 Laboratory of Chemistryof Natural Products

2°

3, Bethesda, Maryland

HHS a Nth

Imet Proj ect ReportVeer 1 958

Chemteoteod Ehrymatio Transformationsof home Qrtoes o

C o C o Sweeter9 Ph one

h e Lereeherg tteehoteel)

Wome n

Petieht Days

prepered eheoteatty hp hydrogen peroxide ortdo=

stated from peper ehromotogreme to pore formo

at orteettoh of oteotthe hp e strain of

s eh attempt to implicate orphteotthe (ore of

edtete to the ortdattre reatttoh ° The Peehooeoees

1 eyhthette eedtoo eehtaththo oteottee as a sourcereete were heed directly or were further separated

ate treettooe o Portteotete freettoes were hroheo

at detergeate o The eoeyoe freettoo which degrededparttehhate treettoh o Experiments to ohteh

orphteotthe es e ethetrete were hepattre n The

Lee after sooto otstotegrattoa or oetergeot

Lete treettoh o

;he tsomers of oryoteotthe with the oetorel

with he mede to order to eetehttsh the eoo=

xteotthe o

Serial No ,NHL- 1 63

slowly dielynehle peptide 9 collidin9 which senses nosediletntion end

hypotension in the dog and contraction of isolated oninea pig intestine o(2) Blood onllicrein is normally bonnd to on lnnotlretor protein and can

not on onllidlnopen end thns prodnee its dilator effect hy moons of

collidln only when the oomplen disassoetntes o (3) Blood conteins an excessof onlliereln innotlnntoro id) Csllidln is destroyed by n blood peptldase o

(5 ) The sellierelns from nrine end pnnerens may he identical9 nherens

these differ from refinery end sernn onllierelns which in tons ore

different from eeoh other

Beonnse no are nltlnntely ooneerned nlth nlend pressore regulation

in men9 pert ienlnrly in its pntholony9 on ere frantionottng homes startingmaterials for the isolntion of the seretel somponents o Hog pencrentie

cellloreln is heing need chiefly to enplore dott led methods o

hoj er effort to helnn ploeed on the isolation of pore onllidinogen

for the following reasons: fl) ln shonld he possible to replete the presenthionssays (sh ines pip intestine and dog preporetionsl hr in ritro enzymaticmethods for the determination of onlliorefn9 innotirstors9 eellidin9 and

cellidinopeno (2) Preporntfon of pore onllldin world he greatly fne ilitnted c

(3) The onestion of the identity of eollidinogen with other o

%e

olohnllns

yielding resoeetire peptides snoh es hypertensinoneng hredy ininogen9and pepsitensinogen oonld he se tt ledo

Erperinentnl

lo Cnllldtnogen° The isoleted oninen pin intestine essay for

onllidinogen end stndfes of the destrnttlon of eollidinonen by pretensespresent in hnman plosmn ore deserlhed in the t orrent report of Dr o h a E 0

Webstero

'

frnotionel smmoninm snlfate preeipftetlon of nhole plosms fontdetednonesh hos heen trled many times nnder dlfferent oondltions o heeorery has

been poor 9 dne in port to losses int erred dnrlnn dialysis o Eren when thiseffect nos mnoh redneed hr the presenee of ndded sonhenn trypsin inhibitorand when this lndihttor nos ndded st eeeh frettionetion s tep9 complete

reoorery hes not heen renllned and the notirftr oeonrs 9 in herring omonnts 9to all the preeipitntes ffron 25 to 50 per sent of setnrntlonlo

heeent stndies soonest the fensihility of nslng ton erehnnners on

e relotirely lnrpe senle to yield e stnhle 9 pnrtfied properetlono Homes

plosmn dilnted firew fold‘

nlth meter end stirred st on dad with h od goreshoot 75 per sent reoorern of nottrity end to per sent reoorerr of non=

dielnrehle solids in the ftltrnte a The snne dllnted plesmn9 nhen stirredwith hE t eellnlose nt oh 70 0 and had? none no notiritp in the filtrate and

sheet 50 per cent in the phosphote hnffere sodinnxohloride ethete 9 sooomponiedby sheet 80 per sent of the sterttng nondialnnnhle solids° The non»

dielrznhle solids in the plesmn nere elmont oompletely ndsorhed by ht

cellnlose st pd 7 0 0 end 8 0 0 nhen the plesmn nos dilnted tnentw oldo

f Diethylnminoethyl

Senin

lo Lebet etery emistnyoi Netnnnl Products

2°

3 ° Bethesdes Mneylnne

PBS a NIH

Inninienel Pnej eet ReneetCelenene Yene 3958

Feet no

Proj ect Title Stenetnne et ennnyllieeeeee A lknlefidno

Frinefieel Ineeetfignten n; G o Wildenng Ph one

Other Kneeeefigetenn S o Uneeg Fnoeo

t o Ennenentg PnoDo

Eltneeetn A : Kfieten «TeennieeBPA ntetnette no Vetneqnen QTeennienl)

CeenenntingUnfiee Nene o

nnn Yenne Pntfient Dene

Tetetg Nene

Pneteeeiennnz 1 0 6?

t

Pnej eet Deeenfietten

Genetnetne enenieel enfieenee nee been entninee fen tee eteeetene

et neennntnteine ( 1 3° enne neeneeente e eeeeee teen the repent ef lest

yenn tn enten en ethylenea innne type etnneteee nee eenefieeneeo In eeeee=

nene nfien eteeetnne ( I) enenentntdtne teens en 0 9 0=efineetnte o En were

eefie (E) in eenneetee te n eenetneny eeeeeeee (EKQ R g enbo ween QII,R2 03) nee eeeeeee eetetntienlfiy nee teen teentee eneeeeetnety nfitn

tnfienyt enleetee nee lieninn ntnnfinnn nyeeneeg e eeneenne ens enteined

enten nee identieel nten einyeneeeenneeentneetne ( 1 1 9 0 teen eennet efien

et nnenenenfietne eetnnltenee tne nnelene efi tee elknleie end leentee the

Serial Ne o

NHI° 1 6&

PBS w NIH

t ienel Ptej eet RepeatGaleneat teat 1 95e

an9 no co g the Stnnetnne et Unenlatineg

at tee neatyllteaeeae o etc the Stenetntee ef

toee ttnanepnenantnetetnetJ; Ann Chemo Seno p 80

no t o » ninetefiee et tee neaeyllteaeeae o XIIO

tee by Seeten and Anni ateenelgJo Ana Chem o See o g

tenet ; B oJo and Wildm8M9 We co g enenaennntnfiefine

inn e Bnteeeneae Nttt egeng J o ,nna Cneno See o g §g9

Serial N0 0NH1 “ 1 66

to very strong eoidence for one with a second possibility not completelyruled out and similarly on the hasis of NMR enidence alone 9 the fifty

e ilre

theoretically possible scrnccnres of alkaloid C hare been narrowed down to

twoo Other considerations may he inrohed to make the final choice ;

although if the NMR stadies were extended to a few other known aporphine

alkaloids in order so get more dale for freonency assignments9 the final

choice conld he made on she basis of NMR enidence alone o The spectrum of

an nnknown aporphine alkaloid seat so me by chemists as the Squibb Companyof A rgentina9 provided enidence lhal the ssrnclnre proposed by the

hrgentinlans was incorrect ; and possibilities were congested9 one of which

can he selecced as agreeing wish earlier chemical eridence °

(3) The fourteen alkaloids isolaled here froml hnnagig anara hare

been examinedo The onhslandinn recalls are she incerpreeacion of the

speclran of lanacrine which enables ins complete sarnccnre so he written9the applicarion of this knowledge to she minor alkaloid lanine which gires

one local slrncrnre decenninalion of chic alkaloid from merely she

analytical dale and the one specsrnn9 she srrnccnre proofs of hydroxy=

lanacridine and hydrorylnnidlne were accomplished he NMBQ and finally? the

posiaions of she hydronpl grasps in hydronylnnacrine and hydronylnnine mayhe assigned on the heels of their Nhh speccrao

id) Sereral scrnclnres hone heen proposed he narions innesrigalors

for she alkaloid conessine ° Only one of chase was in agreement with she

hhh specsrnmo (Schoennenaly9 she parsial synohcsis of conessine was

achiened by Br o d o do Cory which panned ins ssrnccnre s inwas she one

indicaled by Ndhlo

of Ge-l~ - f¢lfi 2’ p d W !

Ascension is being given so preparing one man saddles for

pnhllcarions additional dale nose he ohlained on many of she alkaloids o

Olher aporphine alkaloids are holnn collecred wilh she assistance of Dr °

Innhnshi and Br o B o Schlirller oi’

Ciha° la is hoped that is will be

possible so use this techninne in studying nnsolred problems in the

srrnccnres of orher alkaloidsg sa

g?as questions in siereochenistry of

the onioine alkaloids and in she « yohimhine series°

Part B inclnded

Serial No a“HP -1 67

lo Laboratory of C hemistryof Natural Products

2 0

3 ° Bethesdag Maryland385 a NIH

individual Proj ect ReportCalendar Year lied

Proj ect Title Studies on A lhaloids o To discover new

alkaloids of potential therapeutic valueand to e lucidate their structures o

Principal investigator Parole Z altanan (Visiting Scientist)

Other investigators

CooperatingUnits Br o Bernice G o Schubertg Plant IndustryStationz d o S o Department of Agriculture?heltsrille g Mdo Plant identifications o

has Years Patient hays

total ado None

Froiessional: noOther

Proj ect Description

hinatandra _hei_graueana thinatandraceaei

This plants anonp otherss not sent to as iron Neu' duinea as part

propran oi ihrestigation regarding thee"Harmon

’"disease that has head

up hundreds oi notices at that islands'

The il lness nae descrihe d up Brae Gaj duseh and Z ions in a report

no hen hug nai of Medicine thong 1 957) p in nhich they stated

the onions logic degeneration found on histologic study strong

roasted cone doctor o

A crude extract oi the alhaloids oi the hurt at Hinatandra ahoned

1 strong physio logical actiuitp in rats iconoulsious ending

=

inm

deathlional crystal liz ation and chronotopraphy on alumina produced too nain

aidsg one of nhich n hiuhacine z duct do Guano 9 9 283 lithe) »

”up a p a tool[a] a co s

@

[c soon in cuci3]

Hinhacine retained the actiuity ohsersed in the crude extract

the other alhaloidg mop a 2l?» 223@9 not identified» nos inactive o

The lethal dose of himhacine hydrochloride nos ice on /hgo uhen

run one inj ected intraperioneally in nice o

Serial No D“Em -1 63

lo Laboratory of Shenlatryof Natural Products

2°

3° Bethesda» Maryland

iiridnal Proj ect ReportCalendar Year l958

Syntheciag Degradation and lnterconoeraiona

of the hnnryllidaceae A lhaloidao

hoary Mo Fe leag Ph oD o

Patient Dara

None

the Poet leer

heen node in the interconneraiona at many at the

a and certain ateric re lationships have evo lvedo

e at haenanthanine tnatalenainel has heen toned

rted to aponatalenaine till he dilate echo The

ponatalenaine till has been prepared ironh in tern hat heen converted to tanettiae illllo

at the 3 alhaloida in identical » and neceaaarilycatatao Farther the methanol and phenol gronps

S erial No e“HI “ lé g

derivative (XII) to hoennnthine (XIII) ? the key slhelold on onieh the

contlgnt etlon of all the osonps ln slng C is hoseoo The hydrosyl groups

to slog G ooe‘gis oleslol slnoe e eycllc eenhonote (XXV) nos tonnes on

tteotnent of Xlll with phosgene o

Final ly hnohenenlne hes heen tonne to possess stnnotnne xv

slnee fit was eonsented to the eoosyoso eenlsstloe (XVI) on tseetnent with

ehnonlc sclee

oysldlne folloeeo os slhellne hydrogen oesosloe q This

someones attended othyonoeslneolotne (XVII) with llthlnn olnnlnnn hydslee o

Thls sons conooono nos ohtolneo also by the sotlon of llthlnn slnnlnnn

hyoslee on oolnonlolne o The oonhle"

hono hos been oloeeo tn the honoollyllo

oosltlon heeense of tellers of the hseeonyl stone to oslolse elth nengonese

dlonloe o

Ethe l nlnos points eoneennlng the steeeoehenlstny and abnormalneoctlons of sesesol of the shone slhololds will he lnoestlnoted toothes c

The stnnctnnes of nontonlne one coeelnlne see helng elsnltled end theln

teletlonshlp to hoenonthenlne ls nodes int estlgetlono Longer quantities of

the ootleol nnttoooe of hephenlslne one other defloetloes one being pse=

Serlol mo o“HI- 1 68

lo Lohorotory of Chenlstn§n

of Notnrol Products

20

3 ° Bethesdes Maryland

lndlvloosl Psoj eot ResortGolenoor leer l958

t llcetlons

es» H o Mo and hlloneng so 0 0 9 lntesconnerslons of Anetyllldeoese

elolos by Soelon enn. hosl hloohols lo 4hno Chemo soc o so 4395 a 4404

solo

esp no so and hlldneng h o co g Stroctnse of heenenthenlneg ChenlstsyInehstsyg esta tes lllfihlo

es 5 0 9 Holes? H o h o » hlghetg h oJo one. hlloeen9 h a 6 0 9 Onoheenenthlolne

loyellc Loctoo Possessing o Bridgehese NttsogensJo hnna enema Soc o ggg09 259l ( l95elo

es ; H o h e and hlloneng'

wo co , Stetsoehenlstry of the 5 9 l0h=Ethonoohenm

hrlolne hlhololos of the so one thes e Sos a « In press) »0 l95elo

Serial “HI “ lé ’

initial observations o deve lopedtive components iron oe teriels which indicate

to as observed in the glyeoside testing programo

Serial to 0“ 11 1 4 71

1 0 Laboratory of CheniSiryoi Nainrnl Produces

2°

3° Bethesdag MarylandPHS NIH

Indirional Proj eco ReportCaieaoar

The Tooting oi Piano Memorials forniiaioine and Giyoooioeo o

1 inoeei igaioro maniaLo nopereon» or e

yesiigoioro James D o Link (Technical)oooaiao LoJohnson (Teonnioaii

innUnioo B o G o Sonnberig Piano inonoiry Soaiiongwe 5 0 Deparoaeni oi Aprieoiinre pBeiisoiiieg one Piano ideniiiicaiione

ano Froeorenenio c

Feiieni Bays

None

ieoeripiion

see Dnring_ine Paoi ieag

naiog 3355 piano oanpieop onion incinoee 524 Heroerinn

re been oooooo ion aikaioioo and ooi aaopieo i25ii paceLio n The ooeai nnaber oi pianos aoreened repreoenie an

DO anaiyoeo since Deceaoer 3i9 223 oi anionwere node

:igaoor in Menico o

anoiiion io one aiiaioio oeierninaiionas 54 of one above

seen oereeneo for gipcoaioeo and one eeooiia on 5 more samplesPapoioiopioai activity one observed for 35 of one pianos

aoaocnianeone inj eoiion oi nice °

$ 0

ienoion will be ioeaoeo on a more aianoaroiaeo proeeoure ior

ion oi payoioiopic aiip aoiieeg oaoer ooiopie ; nonwoiioioioai

no eniaiinp piano oappiieo o

Serial N0 0NHI- lWe»

colorimetric method oi‘

li sts and Snbbaror to determinetbeJI.:i.beratedo Most of the dogs used for pathologi c study were

to sodium pentobarbital l=~2 days after a large dose of norepine

epinephrine o ape cimens from nearly all organs vne t's fixed in

.t fornello buffered to pH 70 0 0 Routine paraffin sections

ed ri te hematom lin and eoeina Froz en sections were s’rained

l fat wi th Oil red 0 0 line nd crosccpi c findi ngs were compared

seen in dogs given saline ih i‘

nsniono 0

Findings s in intravenous inm sion in conscious dogs of a

of norepinephrine (G oa-4 30 85 rug/kg) or epinenhrine (06 5°is followed by an elevation in SEE-Ow l whi ch reaches a peak

hours and subsides wi thin dayao SGPe'l‘and serum alkaline

e increase more gradually and subside more slowlyo Pathologic

nfirm the myocardial and hepati c damage suggested by the

erase enzyme levele o Subcutaneous injecti on of 1 mg/ kgproduces less constant and severe pathologic cha'xges9

lowed by a gradual increase during the first day in SGO e'J9

serum alkaline phosphatase with peak levels considerablyn those foll owing the intravenous inim i che o The adrenergic

gent Dibenzyline prevent s the ri se in sewn transaminases but

so in semn alkaline phosphatase if inj ected intravenouslyof 2 mg/kg one hour before admini stration of epinephrine

=i n=oil o

ficanee_

Of the Irw m tei s These findings in

at the possibility oi‘

similar findings in mane Norepiner‘hrine

used to maintain arteri al pressure during coronary shoe}, and

sad di agnostically in myocardi al infarctiono If norepiA‘

lelfi‘

ll"3-Ite

elevation of in mans,the interpre tation of elevated

la aft er coronary occlusion may be corrzfi asingo

fi ndin s suggest that large doses of norepinephrire and

e=in=oil should be used cautiously» because of the possibility

cardi ac and hepatic damage o

sed Course of Progect z This proj ect has been completedo

le e

Serial N0 0“HI- 1 75

1 0 Chemical Pharmacology2° Cell Permeability3 ° Betheede g Maryland

Individual Prejeet Repere

Calender leer 1 958

et Title Entrance of Suhetenees into the Central

Nervous System

ipal Investigator s Dr . Lewis S o Schenker

Investigator Dr e Hermeem Kure

areelng Unite Dr o Kare “ salary was paid by'

e fellewehipfrem. the Mex Kaee Feumdaelon

fears (Celender Year 1 958) Patient Baye s None

afess ionels

Ler 0

see Description

Ob jeeeivegs To determine what factors govern the rates at

g drugs pass from the bloedetreem into the central nervous

amo

Metheds Egglogggs fiege were anesthetiz ed with chloralose and

zen» a polyethylene catheter wee placed in the cisterna magna

she eolleetion of eerebroepinel fluid (CSF) » and ver . ous drugs were

liseered Via the femerel veino Concentrations of drug in the

and in plasma water were measured in samples colleeted at var ious

in The relative rates at whieh var ious drugs ent er the CSP were

ared graphically by pletting the CSP te plasma water concentration

>e against time a

jgfl xr _F1 ndgg; e a The time required for various drugs to reach

luilibrium CSF Sple eme concentration ratio of aboufi 1 00 ranged

several mimetee to more than 5 hours o Some drugs entered the

30 slowly'

thet they were barely detectable even 5 hours after

fining the iatrevenous administreei ea of the drugo

Serial N0 0

1 , chemical Pharmacology2c Physiology

3 , Bethesda, MarylandPBS- NIH


Project Title : Studies concerning the role of epinephrine and

norepinephrine in lipid mobilization and

deposition,

Principal Investigator : Dr e Harriet Mo Maling

Other Investigators in», William Mo Butler »Jr o

are “ Martha A 0 Williams

Cooperating Units Dr ° Benjamin Highnany National Institute of

Arthritis and Metabolic Diseases

Man Years (Calendar Year 1 958) Patient Days

Total 1 025Professional : 0 9 1 5Other

Project De scription

Ob jggtgoggs To elucidate the role of epinephrine and nor»

epinephrine in the mobilizationé nd deposition of fat e The demons

stration in this laboratory of myocardial fatty changes in dogs

after intravenous‘

nfnsions of‘

large doses of epinephrine or norm

epinephrine and the prevention of marked fatty changes by the

prior administration of the adrenergic blocking agent Dibennyline

both suggest that epinephrine may be involved in the deposition of

fat o Other investigators have shown that adrenalectomi z ed animals

do not develop fatty livers under circumstances producing fatty ih

filtration ih intact animals (Ramsey and Goldstein» Her o

Q1 : 55a 1 957) o If epinephrine is essential to fat deposit ion in

the liver and other organs » adrenergic blocking agents would be

anticipated to be effective in preventing fatty infiltration of

the liver by diverse agents o wool and cc-workers (An°Jo Physioi o

lzgs 4273 1 954 ) have sheen. that ergotenine partially prevents the

fatty livers which can be induced by ethionine in fasting femalerats o we are stuiying the effectiveness of various adrenergio

blocking agents in inhibiting fatty infiltration of the liver a

If adrenergic blocking agents » with different chemical structures ,all are effective in inhibiting fatty infiltration of the liversit is probable that epinephrine plays an essential role in fat

mobilizat ion and/ or depositiono

Nonu infarcted mus cle and nyocardium from other reserpiniz ed dogs not subj ectedto coronary occlusion were analysed for norepinephrine content o Myocardial

infarcts were examined grosslyg and fired in 1 0 per cent formalin buffe red t o

pH Froz en sections of the myocardium. nere stained with neutral fat with

Oil red 0 0

Mi :y n Lg s The mortality after coronary artery occlus ion is

approximately the same in reserpinised dogs as in normal doga e Two of lireserpiniaed dogs died from. ventricular fibrillation within minute s after

coronary artery occlusiono This may be compared with lo deaths within 30

minutes after occlus ion in 1 01 operat ions on normal dogs a

Although in the reserpiniaed dogs » the heart was depleted of h ora

epinsphrine g spontaneous ectopic activity developed as usual the day after

coronary artery occlusiono The duration of the spontaneous e ctopic activitywas within the usual limitao Cardiac hypersens itivity could be demonstrated

after the spontane ous ectopic activity had disappeared o These findings indi e

cats that the spontaneous ectopic activity and the prolonged state of cardiaf

hypersensitivity cannot be explained in terms of the release .of norepinephrine

from the infarcted muscle during necrosis and its absorpt ion on the neighboring

The gross and microscopic appearance of the infarcts was not significantlydifferent in the reserpiniaed dogs from. infsrcts produced in non- reservini z ed

dogs o Fatty changes were present surrounding the infarcted muscle in reser

piniaed dogs » just as in the contr

ol dogae

that the amines in the myocardium. do not contribute significantly to

the spontaneous e ctopic activity and the cardiac hypersensitivity fellowingcoronary artery occlus iono

P -@sem _

publication,

of,

Praiagt : These findings are being prepared for

rt B includeds Yes

Serial No °NEE - 1 78

1 0 Chemical Pharmacology2 o PhyS iOlog

3. Bethesda, Maryland


Calendar Year 1 958

Part A

Project Title High Altitude Tole:ance of Normal Dogs and

Dogs uith. Myocardial Infarcts

Principal Investigator : Dr ° Harriet M; Maling

Other Investigator Mrs. Martha A ° Williams

Cooperating Units Dr . Benjamin Highmang National Institute

of Arthritis and Metabolic Diseases

Man. Years (calendar year 1 958)Total 0 0 2

Professionals 0 0 1

Other 0 0 1

Pro ject Description

Ob je ctiregs To _determine whether myocardial infarction impairs

appreciably the ability of’

dogs to withstand a simulated exposure to

a high altitude s

M: hod _m luv eds Normal dogs and dogs with myocardial infarcts

produced by twea stage occlusion of the anterior descending coronaryartery were placed in halters and tied with leashes to the walls of

a large decompression chamber o Lead ll electrocardiograms of some of

the dogs were recorded outside the chamber on Sanborn Viso~0ardiette3 9with all connections made through a hen on the wall of the chamber o

Electrodes were held in place on the dogs0 legs with elastic bandages °

The connecting leads were brought out through the halter in such a

mey'

thet the dogs were able to stand» sit downy and walk unless restricted

by the lssab a The physical activity and respirat ion of’

each dog were

observed by frequent intervals thr ough windows in the walls of the de

compression chamber s

Simulated altitudes of 34 9000 and 389 000 feet were obtained bydecompression within the chamber in a stepwise mannero

Part A

Serial No e“HI- 1 8°

2° Drug Metaboli sm3° Bethesdag Maryland


Proj ect Eitle The Enz ymatio Oxidation of Ni cotine

Prineipel tlnveetigator z Dr e Howard B o Becker

Other Investigator 8 None

Cooperating Unit s NOne

Man Xears (calendar year 1 958) Patient Deye (calendar ye ar 1 958)Total : 1

Professional : 1


Obj e ctivee e $0 study the enzymatic systen(s ) by'

whioh nicotine

is metaboli z ed in animal organisme o

Met hode Engloyeds A differential solvent extraction. method, for

the simultaneous determination. of microgram quantities of nicotine

Megan Findigge s Cotinine was shown to be a major prodnot of

cotine oxidation'

ny a rabbit liver preparati on consisting of

microsomes and soluble frantiono Approximat ely 50 per cent of the

added nicotine is conjnerted to cotinine in this system° we have

previously'

noted that TPNH and. oxygen are required for the reaction9which. is catalyz ed by microeomes Q

A pnroxime tely 1 0 per cent of the dose of nicotine administered

to rabbits was shown. to be excret ed. in the urine as eotinine a Onlya minor fraction of the dose of cotinine administered. wae excreted

unchangedo TherefOre 9 i t seems likely'

that ootinine is a major “

intermedi ate in the metaboli sm of nicotine in vivo o

Serial No o“HI-4 31

In. this proje cts we hope to answer the following questions

(l) Is cardiac hypersensitivity always present when. myocardial

fatty changes are present? (2) Are fatty changes inrariably

present when myocardial hypersensitivity is demonstrable?

methods ggployg Electrocardiographic responses to test doses

of norepinephrine are re corded. before and. at varying times after

procedure s which. might be expected to cause fatty ch anges in. the

myocardiumo the induction of ventricular tachycardias by these

test doses indicates cardi ac hypersensitivityo

Dogs are killed. for pathologic examination. by the intravenous

administration of sodium pentobarbital o Tissues are fixed in lo%formalin buffered.

to pH Froz en sections of the myocardium are

stained for the presence of neutral fat with Oil red 0 0

The duration of fatty changes in the myocardium

nfusione in normal dogs and. followingcoronary artery occlusion corresponds roughly to the duration of

cardiac hypersensitivity o

Exposure of normal dogs to a simulated altitude of ft o

for 3 0 5 hours does not produce cardiac hypersensitivity of signifi cant

myocardial fatty changes9 suggesting th at hypoxia alone is not suffi cient

to produce these phenomena°

Diphtheria toxin does not cause myocardi al fatty'

changes or cardiac

hypersensi tivity'

in. the dog (sim animals )9 even.though cardiac arrhythmias

and. fatty changes lu. hne he a rt have been reported in. man after diphtheria o

‘Marked jaundice i s produced. in. the dog by diphtheria toxino

Intravenous'

infusions of serotonin (in. doses up to hmg/kg) over a

period of about 9@ minutes eith er do not affect arterial pressure

significantly or lower the pressure moderatelyo There i s no cardiac

hypersensitivity the next day o lhe monoamine oxidase inhibitorJB 51 69given daily by subcutaneous inj ection. in a dose of 2 mgfikgp does not

cause cardi ac hypersensitivityo

Mild cardiac hypersensitivity was demonstrated in one dog after

chloroform inhalationo However? liver damage i s more marked than

cardiac damage after chloroform” and it is difficult to produce a

prolonged. reversible state of cardiac hypereroitability with this

agent o Me et dogs either sham no after=effects or die within 21 3

_ _ m

This project may

give insight into the physiological and. biochemical mechani sms nudare

lying prolong ed states of cardi ac hypersensitivityO .An. understandingof these mechanisms may provide a rational basis for the sele ction of

Serial No e“HI- 1 33

Sympathomimeti c drugs vary considerably in thei r t endency to

produce ectopi c activi ty on the fourth day after coronary arteryocclusion and i n their effects on we spord aneous ecfopic activity o

These effeci s seen to be correlated with their effects upon conuractile

force oi‘ ‘the heart

m ficance to ihe i’rom an of the Institute

" This N EW may 5 63

helpful'

i n selectingdrugs for use after coronary occlusion in mane

Pro -osed_ Course of Pro cess The comparisons of arrhyrhmias

induced by synpa h rrimeiti cs before and after coronary artery occlusion

will be condinuedo I'

he sfudy will be extended to include oiher cardi ac

stimulant drugs including seroronin and iheophylline o

Part B includede‘

No‘

Part B

Serial. No o MRI- 1 8h

gggor Findings s In dogs given daily doses of 2 mg/kgJB 51 69the serot onin concentration of the brain. st em increases markedly and

the norepinephrine concentration. remains unchanged o In contrast , in

rabbi ts maintained on the same does the norepinephrine and

epinephrine concentrations in the b tem increase o

Dogs did not show any significant change in arterial pressure

when maintained on 2 or h. mg/ kgJB Slo (in some dogs, 5 days/reek) forperiods of h=20 day

'

s o Even. mhen neurological. symptcms were marked9these dogs maintained their resting arterial pressure during tilting.

Rabbits given 2 mg/kgJB 5io for h. days (1 rabbit ) eud. lO days

(1 rabbit ) did not show any'

change in arterial pressure and. did'

not

show postural hypotension. during tiltings

Three of sin. dosa given 2 mg/kgJB 51 6 daily'

for he l? days showed

symptoms of ataxiao These dogs more unsteady when standing° The hind

legs tended to sink towards the floor s

A ll of 1 2 dogs maintained on L; rag/kgJB 51 6 daily have developed

marked neurological. symptoms involving especially the legs o Five of

these dogs h ave been given pyridoxine (S'

mg/hg intremuscularly ) dailygbeginning with the first day of treatment withJE 51 6 0

Since neurological

symptoms are prominent even in dogs given oyridonine dailyb the toxi cityofJB Sle in dogs is not due primarily to pyridoxine deficiency o

Neurologi cal findingsinclude general irritability; pupillarydilation and. sluggish light reflex9 unsteadiness during standings sinking

of the hind legsg rigidity of the hind legs9 ext ensor spasms especially in

the front le'

gs9 tremor which is most marked during re stg nystagmus (2 dogs )priapism (2 dogs ) 9 and changes in the character of the barking° De spite

a good appetite” some dogs lose weight when given. h. mg/kg daily ;

Neurologi cal symptoms are evident after the second or thi rd

daily doseof h mg/kgJE Sieo One dog died during the night after the

third dose o hematocrit and hemoglobin values were not changed signifi canti:during the first week at this doses but fell appreciably during the

second meek o Serum trans aminase and alkaline phosphatase values may

rise moderately o

gignificance to the Proggam of the Institute s The toxi city ofJB5l6 is especially interesting since this drug has been given a clinical

trial in. the Institute s

Proposed Course of Project s Baroiul histological studi es will be

made of the spinal cord and. brain of dogs maintained onJB 51 69 h. mg/kgdailyo lhe study

'

will be extended to other monoamine oxidase inhibitors o

Part

Serial N0 0

“ 1 4 85

Methods Enplozed: Establi shed methods o

Magor Findiggs s Using a procedure g developed in. this laboratoryher the extraction of norepinephrine from tissues 9 several rabbit

tissues were examinedo In each case the results were inconclusive 9th e only fluore scent compound positively identified being serotonino

A t this stage, a report in the literature on the presence of an

unidentified vasoactive substance in the nasal mucosa of dogs and

sheep led to an examination of the nasal mucosa of dogs ° It was

discovered that there is present in. mucosal extracts a definite

fluorophoric substance , whi ch behaves as a single compound. on paper

chromatograms and counter=current distributiono Its extraction

properties are those of a neutral compoundg and on. paper~chromatograms

it giv es none of the common color reactions ° Also9 it has no biological

activity When tested against the isolated guinea pig ileum? a fact which

distingui shes it from the substance reported aboue o

The fluorophore seems to be concentrated in hhs nasal mucosa

and up to the present has not been detected elsewhere in the other

tissues examined including other mucosal tissues o

An incidental finding of significance was tn'

t serotonin was

quantitatively exiracted. by'

th e method developed for norepinephrine

and. this led to the development of a combined method for the twoaminesg using the one extraction. procedure instead of the two which

had. been. developed. preriously o This will facilitate research programs

which involve the quantitative assay of norepinephrine and serotonin0

characterise

tion ctiveg organ specific

substances will lead to a clearer understanding of both the normal

and. pathological processes uhich take place in the animal body o

Proposed Course of Progec 8 Further investigation. into the'

propertiesg di stribution and biological activity of the mucosal

iluorophore will be carried ont o

includeds No

Serial mo o

“HI- 1 86

placental membrane may'

thes be likened to similar transfer across~

the lipid membrane of the fi sh gill o

flam e ten _

1W g _

the Irw itm s These studi es

suggest that the mi osomal enzymes responsible for the oxidative

metabolism of drugs are furth. r examples of"ontogeny recapitulatimg

phylogeny“° The practical signific ance of these studies is in the

field of therapeutics o The lack of t hese enz ymes in the neWborm

indicates that drugs should be used in. the neWborm. with extreme

g. may explain the texis effects to the newborn child of

some/given to the mother prior to delivery o

Proposed Course of Proge ct s Further studies are necessary in

order to ascertain. more clearly the time of development of all enz ymes ?for the metabolism of foreign compounds including those for 1 ) sulfuroxidationb 2) merC rpturio acid9 3) ester end amide hydrolysisg and h)dehalogenationo It will also be of interest to study the mechani sms

which. oontrol their development and the possible role of hormones in

this controlo

Part B inolededs no

Serial mo o

”11 4 87

with aniline 9 monomethyl=h=sninoantipyrine or be aminoantipyrine o

Evi dence was found for an acid labile metabolite of aniline 9 involvingthe amino group o No cofactor requi rements have been discovered as yet o

In a study of aquatic arthropods such as crayfish and lobster-cgcompounds such as aminopyrine and chlorpromaz ine were metaboliz ed

g vivo o A ttempts to locate the sit e of ensymatic activity by Lgvi tro studie s using the hepato

=pancreas were unsuccessful asthe

pr eparations did not respond to attempts at activation wi th a broad

spectrum of mam eliah cofactors o

A stuw of this

type is important in the productionof data which could produce a

significant biochemi cal differentiation of species o

Prog sed Course of Erog‘

ect s A ttempts mill be made to determine

the speci fic localisation of these enzymes in the arthropods o A t the

time the re are indications that the cofactors involved may be verymuch different from those utili sed by similar enzyme systems i n

mammals o The mechani sms of metabolism of foreign compounds in

arthropods will be studiedo

Part B includeds No

Serial No °NHI’ 1 88

fbreign compounds in toads was indeed different from those in. mammals o

the enzymes involved were found to be locali z ed in the soluble fractionrather than in the microscmes 9 and did not require reduced triphospho

pyridine nucleotide (TPNH) and oxygeno Further9 inhibitors of the

mammalian enz ymesg such as SKF SZ Se A'

were wi thout effect on the toad

enzyme systems o Properties of enz ymes indicates that dehydrogenaee are

involved a whereas in reptiles9 birds and. mammals the enz ymes activate

oxygen from the air °

A detailed study of the metabolism of MMAP and. aminopyrine

marines has shown. these drugs to be metaboli z ed along a different

pathway'

than in. mammals o MMAP is not demethylated. but is t ether

converted to he hydronyantipyrine o On the other hand9 aminopyrine is

demethylated to MMAPQ but by dehydrogenation rather than an oxidative

mechenismoEhe enz yme loses activity on di alysis and. may be reactivated

by addition of TPNO

gnificance to the Program of the Institute s The importance of

this problem to the evolutionary development of animals li e s in the

question. of whether a biochemical function which is solved more than

once in. evolution will be solved differently'

in. each instance °

Proposed Course of Progecta hurther studies on. the enzymes involved

will be carried out o Particular emphasis will be placed on the mechani sms

involv ed in these processes p the co=factors involved9 and the products

fbmmedo A ttempts will be made to isolate and purifly the enzymes in=

volvedo

Serial No 0“31 4 -89

Me or Findi s : Ainphibia such as frogs and salamanders were

found to have glucuroni de conjugating me chanisms similar to those of

higher vertebrates o howeverg all attempt s to produce phenyl glucuronides

in fi sh , gig vino ? or in liver slices or homogenates reinforced with

glucosea were unssnmccessi‘

ul o A stum was made of the missing factor in

fish o Surprisingly enough fish microsome s h ave the glucuronide transferase c

thus im ubation with phenolphthalein and exogenous UDPGA (activeglucuronic acid) result ed in formation oi

‘

phenolphalein glucuroni de o

Howeverg the soluble fracti on of fish liver (789 000) x g supernat ant )was found to be lacking in the DPN dependent dehydrogenase enzy me which

is required for the ofi dation of uridine diphosphoglucose to UDPGA 0

.M .

of tb_

1 1 Mu e 2 The development

of the mechani sms for glucuronide and sulfate conjugation are exceedinglyimportant in the me taboli sm of foreign phenolsg alcoholsg acids and amines »and also in the excretion of normal body constituents such as sterols and

bilirubino the reason for their development in evoluti on would be a

great help in understanding their functiono

Progosed Gourse oi‘

Progect g Furthe r studi es will be carried out

to more spe cifi cally describe th e enzymatic defect in glucuronide

formation in the iish o the mechani sm of sulfate conjugation will bestudi ed in a similar manner o An attempt will be made to determine the

effects oi“

lack of the se mechanisms on the metabolism and excretion oi‘

bilirubin and. sterols in the fish o Animals forms both aquatic and

terrestrial which are lower than fish in evolutionary scale will be

studi ed to see how they dispose oi“

phenols and to obtain an answer as

to whether the enz ymes are in intermediary metabolism im ortant o

Part B includeda No

Serial 1 mo”HI- 1 9°

ion of'

barbiturates 9 Ne alkylamines 9 aromatic eth ersg primaryaromatic rings, fish were found to be totally lacking the

Amphibia are the crossroads of the evolutionary pathway from

terrestrial existence o

s found. that those leading an. aquatic life lack the enzymes 9e amphibia Which maintaim. a terrestrial exist ence possess

ive me chani sms o The repti les possess oxi dative mechani sms

ioh are localised in liver microscmes 9 similar to those of

as While others are locali z ed in the s oluble fraction of the

ilar study of arthropods showed both insects and crustaceat s

to oxidi z e foreign compounds» the fully t errestrial insects

efficiently than partly t errestrial arthropods o

ther enzymes for' metaboli sm of foreign compounds s A similar

is stud y of the enz ymatic me chanisms for conjugation of

enols has led t o the di scovery'

that these pathways are

fish. but present in other vertebrates o Other workers'

h ave

etc to fomm glycosides of foreign phenols o

cults suggest th at the development of enzymatic me chanisms

of many foreign organic compounds in animals parallel

environment » i o e o from aquatic to terrestrial ;

id not need these mechanisms since they were

compounds through lipoidal gills

th emergence onto laud? howeverg there arose a need for

of'

s aterc This was solved wi th a less permeable skip stated the development of mechazd sms to cope with lipid

the Institute s This work explores

d‘

biochemi cal problems in

evolutionary development o It thus represents ah approach

standing of the mechani sms insult ed in flhe'

biochemisthy

curse of her pathways for’ the metabolism of

studied to see if they fit the concept of

f they are is reality necessary parts of

Serial No oNHI ‘ 1 91

2o Bi ochemistry'

of Drug A ction

30 Belhesda9 Maryland

Individual Proj eefi Report

Calendar Year 1 958

Pro je et Tille s Studies om. Meohanism of .A c?ion of

Ergotrepie .A gents

Principal Investigator : Br o 11 °F o Bogdans ki (6 monfihe )

Oflaer Investiga+ore Dr e Fridolin Sulser

we ;Jame s wa¢+eCeogerafiiggUni t s Dr e Suleer on Fellowshi o from ehe Swiss

A cademy of Medical Science

Man'

Years (calendar year 1 958) Paoient Days (calendar year 1 958)

Pro j ect Deseription

Ob jeefiives s Ergetropie agenfie produce an exci+afiory'

pattern

lv ing behavi or, flue em etic system and the sympa'fhem o nervone

sysfi‘

emo Ihese agenls Show struolural similarioies t o norepinephrine

whi eh is poslulal ed as lhe chemical intermediary of she ergolropio

eyelem of Hess o The purpoeee of lhe oresene invesligasi on are :

Io demonslra?e a G eneral sile of eclion in rel e fion to autonomic as

well as behavioral effects and fie employ'

lhe ergoeropie agenfls as

tools for the study'

of oenlral autonomic deoeessants sueh as chlore

promazine ana reserpine o

Methode Egglolg 2 USual procedures used in. pharmaoology o

Megor> Finfi§gg 2 ( 1 ) All lipid soluble congeners of norepine o hrine 9

ae well as coeaineg gives fiypioal ergolropie effeess ineluding excitemenegEEG arouS o l. pal%ern9 increased responsiveness to exlernal stimuli » in=

creased psyehomoeor aofiivifiy and increased cenlral sympashetie outflow0

Since DORA gives lhe same respons es it i s probable thfl t the ergotropie

system is an adrenergie sys+em and may be synonymous with she tefioulo fl

activating eyesemo (2) Some ergotropic agenis slimulaee peripheral

adrenergic receptors as well as central sieesg +hus an effect of

amphetamine on blood pre ssure fie seen. even following spinal section o

Serial No eMRI- 1 92

l o Chemi calPharmacolog2° Biochemistry of Drug

3. Bethesda, Maryland

PHSo NId


reject Title : Pharmacologlc Mechanism of Re serpine Action

in Brain

incipal Investigators z Dr . D o Bogdanski (6 months )Dr . Fridolin Sulser

>operating unit s Dr o Snlser on Fellowship from the Swiss

Academy of’

Medical Science

an Years (calendar year 1 958) Patient Days (calendarTotal : None

Professional :

Other : 0

ojact Description

Objectivgs s Reserpine has been postulated to not central]

stimulating through serotonin, a neuronal system (trophotropi <ich integrates the parasympathetic system with somatomotor and

ic functionao This system seems to act in opposition to a

pic ) which integrates the sympathetic system with

nd psychic functions o This latter system is an

energic system. which is antagoniz ed by chlorpromaz ine a

If this conception is validg then reserpine p in contrast

chlorpromaz ine » should increase the parasympathetic output frs

central nervous systemo

Methodg Emglogggs Usual methods of classical pharmacolo

Majpr Findings : (l) The apparent decrease in central sym

thetic output following reserpine is not central in origino 1 1

5 been satisfactorily explained by the depletion of norepineph:

peripheral nerve endings . (2) Pupillary constriction induced

serpins in rabbits and cats is entirely due to central parasym

ohetic stimulationo (3) Lacrimation produced by reserpine in

bhita and rats is a central parasympathetic response o (4 ) Therasympathetic action of central orig

Serial N0 0NHI ' 1 92

PBS- NIH


Calendar Year 1 958

yards ; and Publications

than abstracts from. this project

tor , S o , and Shore , P . A Interaction of Drugs

a in the Brain. Pharmacological Reviews , in press o

m le Technique Involving Solvent Extraction for

lorepinephrine and Epinephrine in Tissues °

riews , in prese t

aeraction of Psychotrop ic Drugs with Physiologiczhanisms in Braino ‘Modenn Medicine , August 1 , 1 9589

sbache H g , and Brodie » B , B A ssay of Serotonin

itesg Enzymes and Drugs . Methods of Biochem.

op , D °Jo ; and Shore , P . a An Interpretat ion

chotropic Drugs . Postgraduate Medicine , gg, 1 958 0

here , P . A On the Mechanism of Action of

Book consisting of proceedings on the symposia

1 Treatment of Schiz ophrenics given at the Second

3 of Psychiatry in Z urich , Switz erland, 1 957 0

ki , D ° F o g and Shore » P e A Biochemical and

tation of the Action of Psychotropic Drugs o

1 Concepts of Psychosis ," McDowell, Obolensky)

Odie , B. B o : Influence of various Drugs on Serotonin

n Brains Chapter in book on PsychotrOpic Drugs ;

ting t o this project : None

20 Drug Metabolism

3° Bethesdag Maryland

Individual Project ReportCalendar Year

3 Model Enzyme Sye ieme in the Study of

Drug Metabolism

estigeizors Dr eJames Ra Gillette

gator s BiroJemee V a Dingell

(calendar year; 1 958)

A w hen of foreign compounds are oxidi z ed bylob are lo cali z ed in {the m

icroeomel fre efi on of

howeve r

9the neeheniem involved in ehese re

oodo The present studies were

odel systems to determi ne possible mechanism

etiorigo

Eei' e‘

olieh ee’

lmeihode o

A s prev-low ly reportedg +he deallrylaeion of the

es of aniline and h=eminoentloyrine has been demone ereeee

whi ch function i' hm ngh peroxidaseg and ofi deee end

ohenieme o

me oeo be converted to lee sulfom de

deee on dehydrogeneee mechanisme o Both

formation of a red colored

Serial N0 0

The currend studi es have indicated th at tetrabenaz ine inhibi ts +se

reserpine effects by virtue of blocking its acti on. em serotonin. end not

norepinephrine 0 Tetrabenaz ine does not effect the se amines in the

peripheral tissues 9 pla teletss intestine or heart but doe s cause some

release of cat echol amines from the adrenal glands ° The latt er effect

is probably'

medi f ted cent rc llyg The peripheral effect s of reserpine

are not blocked by tetrabenseine o

The short a term biochemical and pharmacological actions of tetrabenaz ine

are relat ed to its half e life in the body which is only 30 minut es o This

action may be contrasted to the longc term effects of re ser pm e which persist

even after reserpine can no longer be detected in fine bodyo Therefores, the

effects of tetrahenaz ine are readi ly reversible whereas t‘

zose of reserpine are

"irreversible o

;_nifi cance to th me em of This oroj ect is an

extensionand continuationof a long- term program concernlng the role of

serotonin and norepinephrine in brain f‘

am c’crtiono

Propos ed course of Progec 3 The current project will be continued in

an attempt to elucidate further th e biochemi stry of brain functiono Studies

of the biological distribution and metaboli sm of tetrabenezine are plen'medo

Part B included: No

Serial No ,NIH - 1 98

By using a new and very potent amine oxidase inhibitor »

phenylisopropylhydraz ine 9JB 51 69 a very marked and rapid rise in

rabbit brain serotonin occurred, indicating a rapid turnover of

this substance in brain (halfb life about 1 0 minutes ) ° Norepinephrine

levels rose more slowly, indicating an apparent halfe life of’

about

3 4 hoars e Central excitation and synpathomimetic responses again

occurred when brain serotonin and norepinephrine levels reached 2

3 times the normal value o

In cats or dogsg honorar y no central lexcitation could be

observed after iproniaz id orJB 51 6 treatment o In these species »

brain serotonin levels rose rapidly, but little or no change in

norepinephrine concentration could be dete cted ° These results

would suggest that the elevat ion of brain norepinephrine levels

but not serotonin levels is associated with the central «e ffects o

n attempt to learn more of the

meohanism. of action of a type of drug demonstrated to be useful in

the treatment of various cardiovascular diseases , Furthermore » theymay help suggest normal roles for serotonin and ncrepinephrinea

substances implicated in the function of home ostatic mechanisms in

gpp gpsed Coarse of Prggect s Further studies will be carried

out in an effort to determine which enios a serotonin or norepinephrine g

is of maj or importance in the action of these drugs , various new types

of monoamine oxidase inhibitors will be investigated o

B included: Yes

Serial No o NHI° 1 99

1 0 Chemical Pharmacology2 ° Biochemistry of Drug Act ion

3. Bethesda g Maryland


Project Title : The Metabolism of Amine Uptake and Binding in

Blood Platelets

Principal Investigators s Dr . F . Barbara Hughes

Dr e P . A , Shore

Other Investigator

Cooperating Units : Dr e Hughes worked under a fellowshipsix months with Geigy Pharmaceuticalu and

six months with Ciba Pharmaceuticals

Man Years (calendar year 1 958) Patient Days

Total :

Professional s

Otheré 0

Project Des cription:

Ohgegt‘

ivsg: Accm nlating evidenee points to an essential

role for serotonin and norepinephrine in brain function" he

amines are said to be “bound“sinee they are stored in

'

a fhrm

which i s protected fron. ennynatic destrnetiono The question

arises as to the nature of the binding and the meeharisms'

whioh

hold and release these ominos o The finding that the same Raunolfia

alkaloids that release serotonin from brain also release it from

platelets suggests that the snhstanoe is held in both tinsnes bysimilar forese e Thus platelets may serve as an in vitromodel

for studying storage and release of serotonin and catecholamines o

Major Findinggs Eoidenoe has been obtained shiel suggests

that serotonin is held within cells by a special meohanism. which

maintains the amine against a concentration gradient o Reserpine

appears to not by interfering with this “pamp o

m By tt mechanismg

reserpine can releas e endogenous serotonin from cells or can block

the uptake of added serotonino

NHI- Z Ol

result s indicate th at previ ously developed

es for the estimation of tissue histamine may'

nd include substances other than histamine o

M ,

2 Because of

ifects of histami ne onthe cardiovascular systemgor more knowledge of the factors involved in. its

gee of Project s A fter perfection of the fluorometric

to tissues9 it is planned to launch an investigati on

esis and. metabolism of histamine and the effect of

its metaboli sm and. physiological actions o It is

s project will be expanded. in the near future o

Serial No 0

nore complete seve ration of pe ti pheral and central effects was

Mb 80 31 1 8 (Ciba) o This carbethoxy syringoyl substitutes

reserpate in daily doses of 50 70/n depletes heart norepinephrine

its and dogs without affecting levels oi'

brainamines o Animals

sedated9 but exhibit hypotension9 bradycardia and a markedlymed response to occlusion of the carotid sinus , Furthermoreg it

the sympathetic responses to adrenergic stimulation and to“

sic stimulationo

is concluded from these results reserpinea like compounds do not

sympathetic by central acti ong but by pwipheral depletion of

sphzi ne whi ch render sympathetic organs incapable of responding

aliminary studies wi th reserpine indi cate when the drug is given

only a small p ercentage enters the bloodstream 0 It is pos sible

a diffi culty in adjusting dosage in hwe rt ension so th at patients

leo depressed9 is due to the variability in the degree of

ism in the gut A ttempts arebeing made” in collaboration with

9 make a more stable reserpine analog, perhaps 293,

an estezc 0

such a compound will be completely absorbed and wi ll depress

ressure in doses that do not elicit sedationo

epinephrine depletion does not acconnt for sedation and other

effects of resexpine o Thus ? SU (Cibab a deme'rhylamino

ethyl reserpateg almost completely depletes brain norepinephrine

ts when given in do ses of 0 0 5 mg per kg}, but has little effect

serotoxn’

tno The anim ls are not sedateds but 2 mg of drug per kg65% decrease in brain serotonin and a defini te sedation is evident o

from sedation coincides wi th rise in serotonin levels despite

nomepinepl'

xsine o Thus SU51 71 is a valuable tool in

in brain serotonin rather than on nor-epinephrine

factor in the'

central effects of nesezmofme o _

‘l‘

hi s

eeis th at reserpine affe cts a neuronal system for“

ormone ; in cont rast to chlcrpromasine which affects

of the Ins ti tcte z The study of these

the understanding of the hormones

echsni sms and may lead to the develops

ions therap euti c agents for th e control

to continue the ccs’selati on of pharmacological

all changes occurring in the brain and

the me tabolism of these analogs byan at tempt to correlate changes in

th observed responses , In additiong applycompounds ami ch have only p eripheral or onlir

effects are more predi ctable in clinical

disease o

Part B

Serial Mo eNHI ‘ Z O?

experiments on the permeability'

of cell membranes have sugge sted that

membrane s in general are lipoidal in nature since fat a soluble sub :

stances penetrated the cell more readily than fat=insolnble snbstanc eao

The present study has dealt with the relation between the lipide

solubility of drugs and their rate s of absorptiono Io eliminat e the

variable of the degree of ioni z ation of various drugs q a large groupof weak organic electrolyt es which would. be uni oni z ed in the int estine

were selected for the strd'

yo the relative rates of absorption were

compared. wi th the lipidenater parti tion coeffici ents of the uni oni z ed

drugs using chloroform or heptane as the"lipid

"

phase o

The resul ts indicate a rough relation'

betneen lipide solnbilityand. the degree of absorption of a drugo The compounds th a t were

rapidly abs orbed had. relatively'

high lipidm eolnbilities While slowlyabsorbed drugs had. very low lipida solnbilitie s o

These results suggest that the intestinal. nncosa is lipoid in

character ° Thus for the most rapid absorption9 a drug should be

largely unioni z ed in the intestinal lumeng and the unioni zed drug

Significance P9 or the Institutes The se e+udi es

should serve as usefnl guidelines for the synthe sis of new drugs

which will be highly effective when admini stered'

by the oral route c

Proposed Course of Progect s ( 1 ) Investigate the absorp ti on of

quaternary ammonium compounds o (2) Investigate the possible role of

chelation in the absorption of certain drugs o (3? Investigate the

absorption of certain antibiotic drugs which are only moderately well

absorbed in therapeutics °

inclededs lee

Serial N0 0NHI° 208

1 0 Chemical Pharmacology2 0 Cell Permeability

3 o Bethesda, Marylazd


Project Title Absorption of Drugs from. the Colon

Principal Investigator s Dr ° Lewis S ° Schanker

Other Investigator Mr. Panayotis A . Nafpliotis

CooperatingUnits

Man Years (calendar year 1 958) Patient Days

Total s B/hProfessionala l/hOther

Pro ject Description

Ob jectives : To describe the characteristics of the colonic

mucosa which govern its permeability to drugs and other substances o

Mgtgodg Emplogeg: The colon of the anesthetiz ed rat was

cannulated at the cecal and re ctal ends » and perfused with

solutions of various drugs a The degree of absorpt ion of a drugwas .determined by measuring the decrease in drug concentrat ion

after a single passage through the colono

Major Findingg: Earlier work on this pro ject indicated that

the degree of absorpt ion of organic electrolytes from. the rat colon

is related to the dissociation constant of the compoundo Thus weak

acids and bases were , in generala readily absorbed while stronger

acids and bases were very poorly'

absorbed o These observat ions suggested

that the colonic mucosa is preferentially permeable to the unioni z ed

form of a drug and that ions penetrate withw difficulty o The present

report describes atedies which support this working hypothesis and

which. suggest that the colonic access is lipoid in charactero

The absorption of acidic drugs such as salicylic and benz oic

acids was greatly enhanced when they were perfused through the colon

in an a cidic sdlution (pH Aha In contrast » the absorption of basic

drugs like aniline or quinine was decreased at pH 4 , Thus a change

in the intralnnenal pH which increases the proportion of unioniz ed

drug facilitates absorption; conversely » decreasing the proportion

of unioniz ed drngdiminishes the degree of absorpt i cno

Serial N0 0

NHI° 2O9

2 0 Cell Permeability30 Beihesdag Maryland

Pro ject iiile The Secretion of Subsfances into Bile

Principal Investigaier z Dr o Lewis 8 0 Schanker

Coop erating Unii z In collaboration with Dr o 0 0 A drian

flogbeng, Dept o oi‘

Physiolon The Geosge

School of Medicine 9

Nan Ye ars (calendar -

year Pa+ient Days (calendar yea?

Professi onals

Proj eci Description

iecpives s E0 describe ihe means by'

whi cb subsi ances pass fromF — O-A G-p — m v

bloodstreafi’

inie th e bile n

The bile duci of flhe ane sthetiz ed rai was cannulai ed

edicles were ligai ed q A fier an iniravenous inj ectiond sacchamideg bile was collected for 3 to S

'

hoursg

The radi oaciivi iy'

in bile samples was compared with +hat in plasma o

Eindi§g= z Previous week on ibis pro j eci di sclosed ihat

arge lipida insolmbie saccharidesg inulin and sucroseg are secret ed

into bile in significani consentraiions o This sugge si ed a significant

porosity at some locus in {he hepaiObiliary system0

In she work now zreporiedgihe biliary se creiion of immling sucrose

mnd. a smaller mole cuj e g mannitoly were examined in. more de+sil and the

disiribueion of these subsi u nces in liver and muscle tissue was also

determinedo The resmii s are summarised in the fellowing table o

Proj ect(

Repori

ar Year 1 958

ation of Drugs into the Piini iary and

Glands

Drc Cedric'

W,M, Wilson

Dr a Lewis S : Schanker

Wilson ’s fellowship is from inc Medi cal

earcb Council of Greai Britain and is

>nacred from Eli Lilly ,

er 1 958) Paiient Days (calendar year 1 958)

aiionship of the pineal and piiniiary'

glande

'

Drugs are adminisiered iniravenouely'

to case ,

Fieso ai various infervals and. poriions of rhe

surement of drag coni ent c

When snlfagnanidine was giv en intravenouslyint o inc picni iary and pineal glands and

poeirama as readily as inie peripheral tiss ues \

areas of The brai ns such as rhe cerebral car eer

ficulty o

xy ibis laboratory'

demonstrated. thai the central

arai ed from the'

hloodsiream by a lipoid~like

is finding chap ihe pituiiary'

and. pineal bodies are

airaied. by a lipida insoluble 9 foreign. subsi ance

ooda brain barrier"may be absent in fhe se areas

onsidered io‘

be p art of the central nervous

s ihai these areas are not p arts of ihe braino Since

Serial VO , NHI — l

1 . Heart

20 Chemical Pharmacology

30 Clinical Pharmacology

he Bethesda, Maryland


t Title Intravenous Anesthetics o

al Investigator : Dro Peter Go Dayton

nvestigators : Dr eJad e Burns

Mrs 0 Dolores Teller

ting Unite : In collaboration with the Department of

siology, College of Physicians and Surgeonsg Columbia

ity9 and New York University, Research Service ,iel Hospital9 New' York, NGW

'

YO Pka

(calendar year 1 958) Patient Days (calendar0 28 year None ,

ssionel

are None

t Description

ctives a A study of the physiological disposition and

tebolism of various barbiturates is intended to

ierive fundamental information concerning the pharmacology of

Lntravenous anesthetics o There is a need for a potent

Lntravenous anesthetic which may be used in surgical procedure sJf long durationo In this re spect” an effort is be ing made

to find a none barbiturete anesthetic 9 since it has become clear

that barbiturates es s class are slowly me taboliz ed and exert

1 hypnotic and not a truly anesthetic actiono In addition,aarbiturates hove other drawbacks (laryngospasmg depressedrespiration, poor muscular relsxstion, ganglionic blockade 9 e tc

’atient. Materiel m Noos e

site and mechanism. of formation of the

nd the manner in which foreign substances

anter and leave it are controversial matters e The barbiturates

anter and leave the brain at different rates in. pr0portion to

abe ir degree of lipid solubilitye It seemed reasonable to lookfor differences also in their rates of passage into and out of

the cerebrospinal fluids So far only barbital has been studied

intensively in this direction0 (Barbitel. ee s chosen because it

iistributee uniformly in total body water) o Experiments

urrently in progress in dogs sugge st that barbital passe s slowly

Tram the bloodstream into the brain and thence into the core

(Attachment 2)

Serial No ohim - 21 1

Honors9 Awards 9 and Publications

Publications other than abstracts from this proj ect

Marks Lo C o gBurns ,JQJW Bren-zns Lo ,

Caumporrnn'

xe s,3 C L , Trousof, N0 9

Peppers, E omo end Brodie , Bo Bo : The passage of thi obsrbiturete s and

analogs into brain, J a Pharmo and Exptl o Them e , 1 23g

Honors and Awards relating to this onoj ect z None ,

Serial N0 0 RBI- 21 2

a CiP9 unlike 6e moscoptopurins does not catalyz e the

tion of inorganic sulfito by milk X0 9 andg although

of'

hypoxsnthins oxidation by KO9 is complstsiy withoutthe hypoxanthino c catslyooo

Wsulfits oxidssoac systsmo

iqus feature of'

6=ClP as comparso with other known. x0

6=81 P6 8=clh '

oos proposed from, hypoxsnohins o 8=clh byaros e Using tracer dosages in adult rats” the followingboon obtainso to date

rocovcmy experimentsa over 90% of t otal isotope wasin urine” expired 002 coo tissusso

reveals tho prosomso of four maj orhave boon identified as starting

and uric acid o Thsss thsoo products hsvo been estimated

iy by precipitatiOh'

sith unlobolso corrior, and correspond

to 629 21 and 6 S% of total usinsmy excretion of isotopo o

Tissue distribution of total isotope has revealed no marked

listributiom when difforoncos in mitotic rats are taken into

’rsiiminsmy ssposimsnto hows indicsioo o small oxtshs of

,oh of isotops into purified nucleic acid fractions o

,s to tho.

Psozhsmhof _

thsu

lnsoituos a This proj oot extends

s o f?“

rin g mo aw fi'

s mg‘oho uhdsr say for soms

ais laborotomyb to compounds which ass structurally vonynormal msoaboiitoso It is hoped that systematic study of

°ism of such compounds by so called “normal“ onzymos will

atifiyimg enzymatic processes which arc invoivod in tho

.he drugs smo in the development of drug ossistahcs o

>urso of Proj ect a To refine techniques employed io . in vivo

DV8 {0 iasnfif§ all mstsbolitss of 6o clP undor condifionstraces and thosspcuiic dosages are cominstorod and find

itiou, if says in the mods of metabolism. in various tissuss o

19m of the possibis roplscsmsnt by 6a ClP as such» or'

oy cos

zhuhai metabolites of the normal purine constituents of

.os and comfoctors will ho mods” s no tho effects of thsos

zbstsnces upon cellular procassos stuoisoo

PA RT A :

sfirial No NHI° 21 3

1 , “wat t

20 Chemical fihatmaco logy

3° Clinical ?harmaco logy44 , Befihegdag Maryland

PMS-Nlfl

Emdgvaflmal Projgc z RQpO t fiCalemdag Y@@z 1 958

Pwaj ecfi Titla Smmé fiam Imflmced EmzyMQ Symtmaaia in mammalm,

Eriacipal D3 0 Allam Commey

Otham Iavgfi tigacomaa N@m@ °

Cwoyaratimg unit : §®me o

E@m Yeara {calemé ax yggx i958) yagagwfi Daya (calendar year

Ebgmfi ,25 Name ,

Pmaffigaiomahz

0&h@xz

Pro j ecfi Deacriptiom

Tfi agmdy aha effiecg of 9&1 ycyglic hydrocagbons

on the induced aymzhggfig of vaz ioug livaz mfic togomal emzymgg

which fixwga.

GB) in é rug@ 0&m $mdwce the

gymthaaia of wficz ogomal @mfiymaa wfiichsmagabolize foraiga

com@ommd@ .

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of cggtaim pe fiyeyafiic (so mgthynchQLamzhxemeg 39 é ~

bamapyreaeg amé t@ mats wail maxkadlyguer@as@ aha actflvixy @f Eflvez miczogomas mm? 1 } hydgaxylazgBpa

-bamzyyr@m@9 2» m Qmethylate wmfim@aao dyagp amd 3) to reducg

thfi amo limk&g@ ofi amimwamo fiy®® o emzymfi myatemfl axe

aiwilam fio mamy é mug gmgm sp fiwr thgy arg localiz ad

am iivax and t eqmige f?&a mad omygemo The pogmibilitywag investigafie é that 39&

~bemzpyt@me may alga increaae the ac tgvgzyofi which m ga. it was fommd

that this byflzwcaxb@m markedly aha activity oi the

micxoeomal amaymg aymagw'Whicmhydroxylacea ac@eamilide and Plaxin o

Hawav&z a Sgé-bfimgpyr@m@ hag a lgeaex effec t on the emzymaa which

hydzaxylate qufim@ 1 $m@ 9 maphahalamgg @m& pam&fil@xo Lit tle or no

effect wag o QEvad om 2&e aasyma ayatgmg'whach demgtbylace No

m@t&y1 ami££m@ &m@ momOMQahylo é - amimcamafipyxime at which oxidi z @

p amé chloxpz®ma3fime . atmdieg poiat out the

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tie$ kyé x@xylati@m of @@V @Eal 8&bgfixmfies axe to

Set fial m ,W E E -fie»

1 , Heart

2 0 € h@mical fihawmaco logy3 , Clinical Ebammaso figgy

Ba ham a, M mylazmd

EfiS- Nfia

Individwaawfioj ect agwomtGa1 @mdas Ya 1 958

Ero j eca fifiale s Stwdfieg

Princ ipal we ,JDJ, BuxmsGabe ? Emvesgigagaz aa

'

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mm@8 Dnlaz ea fiaglmr

mfigwel $amdmau

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‘

fiamma Siwai waw s k and. G@ igy

Eflh®z agoxiewg Baa®lp Swagz @z ia&é o

Mmm flageg ©ca1 amfia$ yeax 1 958) Eati®ak waya (calendaz year 1 958)Tamal 0 23 mbm@ °

, 2%

0&h@z z

messaiggfiom

Phemylbmgaz©mfi g symahe z ic fiarfivativap

h&s fi®®ad @9@ ia emfi exeazmgme

dia@aeea, W® ghmfi fihgesaawqfiabfia fi® ahaa @f s

maekaé wfi gofifimgg baa

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will figaged 1 m yaafiamas axtht itimo

Eaaient fimmez fial o fibme o

Serial mo o NHPatient Nat@? ials Nene o

I 21 6

“

75W "Cardiotm ic factors isola ted from b mf M a m.

ma 0m m a ‘t mam and blw d 13mm mam a to baaacidm lipidmPram papa? chrom togm phice dam 13m beef heart substanw appeam

m eonmm 1 8 m 20 carbem M am e W arm and ultm vwfilm9 98mm mama-w an unsaturate fflg bm nchm chain strm tm which

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straphanthidm o

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Pagsibm m lgtiam mm mm gn fim gwlatmm m cm im and

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m M dmwm m the abwm a tame acid fm m extm cm of

m gbbm amrthrwytag has M en ancmmtem de This fam e? M am as

wfifim m M m m ible cm‘iwm twg M ”aha mm in a. m y

m minism m a? thee. m um afi’Qapgzaim o

S

fifiudyfmg smwm mm fi a em fim wmm M im te M Mm m sfim g”aw l

m i wim s M y cem m rm awimw’ilg im wm m 1 1mm mm when

gu nm e with a. w ww M fm m an mm m lw m mm cmm o The

«9 ??c m em pim s cmfim cm my m m med hem

m fi efi tm M m m ce a? lipidfi) m pm m ga of

W augh m mmmw m we; wm‘

km ctim pm m im o

Thaw afimiee shm lfi fih®r®fam nm vidm

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Swrifll N0 0NHI~ 217

1 °

20 Chemical Pharmaeology

30 Clinical Pharmacologyho Batheadap Maryland

m


Pfimjgcfi Titlaz Studiwa on the Nataboliwm of St®rdlao

Principal Invastigators s D35 Elliott Schiffmmn

Dro Elwood 0 0 Titufi

0mm? Immatigatom None 0

Han Yaara (ealandar year 1 958) Pafiiemt Days

Tbtala yegr 1 958) sProfessionals 1 0 25GEM ? 3 None 0

Project. D@scrinti@m

marindbuflagin. and. mar1 nobufbtoxin, in the toad Euro marinnao

Cholestarol and stigmasterel are metaboliz ed to

unknown steroide of incrwaaed p olarity by Tafirahymana

fiygifq$migg protez aano Thia convarsion may Se 0? conaiderm

'

m ii00 femica1 Qignificance g si nce th@ grawth inhibiting@ff®ct@ at carfi in0 3tatig paring antagonists upen thi$ organism

may be faveraed by the pragance af choleatswol or flfiigmastgrol

in tha mmdiumo

fietbod@ Emblo; ad a Mulfiipla ©hr0mat©graphic techniqusa hawmera igolation anfi nurificatiem, of compoundao

Varioufi moflifications af radiowautograpuy hava been

amplqyad im. the assay of th@ raeuitg of twacer p arimants o

Patiamt Matarial fione o

“ EL“

? a Rasearch has nwocaad®d in twe diractions

( 1 ) A tfigmptg tn fflmfl afficientlyaynfihesiZ ® tbs eardiagtonic steroié go

3 ) lg viva expgrimentgo

fi®f 0 1

caused

fin imcornoration of iaotope into the sterolagbut not into the cardiac active aubatanseao

Serial No e NHI=21 7

xrso of Fro

§set m Tho attompt will be continued to synthesiz e

ate s ro ,

'

which could be an intermediate in the biological

3 are in. progroas to dotermino the nature of the cholestorol

in tetfiohymomae

soarch for systoms that synthesiz e the lactones efficiently,iaotopio fiQsitosterol injected into the toad will be followedo

to ho prasaut in parotoid which contains

Serial N0 0 film- 21 8

cture of the activating mechanism is obscurc o

t appear to involve so lubilization of the cubstratc glecithin of solublo egg lipOprote il (which isif other phospholioasoc) is not hydzzolvz od by this

the abcchco of cctivatoro

m — a

w,

“ co cctw m om cm nbc of

DW hove» pm vcntcd its idontifioxaim in anim l

t is possible that this: cnzymc may be of imp ortance

abolish of mammalian phocpholipids ,

of the Institutfc u Lack of

came o? Pro cot a Further ctudico of the mechanism

p cop d l inositol will 36 carried ont o

or the am m o and a study of its role in w h en

M. be unflat t cm

Indeed

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Ram a

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m , A R M Gam ay

Ogbfl fi’

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M M Cm abm gamma

M m mam Mam a}

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m agmas: Mam wwl 0 mm ,

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am m o

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in the m g m g m m 5m m {Eflmmg m m Do galacfiom n-cl

cmmfijwfim bly M a m wm w m w m3wm gkm wm mm amid m m

m m m m m ID- gmw m m nwfim o

PBS- NIH

Imdtvidual wz o j acg R@@ormCalendar Yeam 1 958

PW@ j @c@ Tis1 e z Saudiea with. mm@cu1 @x R®1 az amts o

Primicpel Emvaatigaaoxg De g A 1 1 am. C0mm@y

Other EnvggzfigatO fi s : mmgg Nata1 ie Ttowgof

Dr eJG J . Hmrma

C0 @@egafiimg Bait : New Y@zk mmivgwafity Reaeagch Se

G® 1 fiwate z fiwmwz ial fica9 1 $a1 9 Ngw YQrk, m@m Yb :

Nam Y®amg {ca1 emdax y@am 1 958) Eatiema Day@

B

Pgofeaafiomalg 0 36

0&a@r , 33

Pro j ect magaz iptiom

maneuvm $ w mg m two lwm inn mm of

wmscmlar of the bemz oxaso le aeriegp

1 &1 1 ent MMfi®I 1 a1 Nfime ,

Enj or B1 m@1 @g&s F 1 ®$ 1 n ma® beefi usad fox th@

year@ a@ a mwacm1 ax t glamamt firmg which 1 5 tm<cgmtxally lik® mgphgmea1 mg fhg dzmg 1 g camve z

a me tabalite fi©kmed by gwbgaitutien af a hydrc

@h@ amimo atom? “ 1 1 1 8 wgmab0 1 1 tg mam gofiemt e

acaivityg amd it hag xgcently bgem intz @dwced

aa a new dmug, Paraflexo Furthem studiea have

wage s saute ©f fiatab@ 1 1 @m o f bath fi1 exim and 1

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of each comyoumdo Thg overall ach@m@ figr the

and Faxafl®x 1 g f@ 1 1 @wm

E 1 é miw

Hydz oxy- Elaxim

1 serial mo o max- 222

1 ° Heart

20 Chemical Pharhacology30 Clinical Pharmacolog

y

ho Bethesda, haryland

PHSm NIHfidual Project ReportFalendar

‘

Yeer 1 958

tudie e with. L=Xyluloee o

.gatorz DroJohe .Jo Burns

or; ereJulian Keefer

:z Studios corried out in collaboration with Dr e

[ell of the National Institute of Arthritis and

.eeaeee o

tar year 1 958) Patient Days (calendaeyear 1 95833 None o

To study the origin and fate of Lexylulooe s the

ed by patients with e ssential pentcsurie o

rial None .

o Previously we described an active enzyme in

hich decerboxylatee Le gulonic acid forming L«

be enoymees Le gulonic acid (DPN) dehydrogenaso hao

ified about BSG fold from hog kidney and a study of

ee wee underteken° Evidence has been obtained for

.onic acid as the intermediate in the reactiono

the Progrem of mo m en tum a LoXyluloee is

f .; fmetabolism. preeenk

wee of Progecta ( 1 ) In the course of the isolatioo

Wfibolic product-

wee uncovered which has

tifferenm proportiee from any known pentose o The

.cm of this compound one its mechanism of formation

iedo

(2) A omell amount of Le ascorbic

terved to be formed from. La gulonic acid in the kidneyt possibility that this may eeflect a different‘

Lo eecorbic bioeyntheeie than that found in liver

retigetedo

Serial N0 0

1 a Heart

2° Chemical Pharmacology

30 Clinical Pharmacologyhe Bethe sda, Maryland

PHS o NIH

Project Reportor Year 1 958

ice with Inositol o

.ors Dr ,John.Jo Burns

Miss Natalie Trousof

Miss Carole dvans

Dr o Nicholas Papadopoulos

‘

his study use carried out in collaboration with

Agranoff of the national Institute of Neurological

Lindness o

year 1 958) Patient Days (calendaryear l958) g None

investigate the precursor role of inositol for

Le of Dmglucuronic acid, L=gulonic acid and La

11 a “0 138 0

Evidence has been presented by others that

averted to a racemic mixture of D 9Le glucuronic

in rat kidneyo Since Daglucuronic acid can

ursor of Lo gulonic acid and Lu ascorbic acid, the

al in this bifisynthetic pathway was studied with

i inositolm c o The results obtained indicate

is converted in the rat to Dm glucuronic acid

acido Since pre vious studies have shown that

ids are degraded in the body via Le xylulose it is

let oentoses play an immortant role in the

Lnositol o No conversion of inositol to Le ascorbic

ironic acid was detected in the rat o

Institute a Inositol is a

the brains heart 9 liver and other organs o

a of P roject a This project is comple teda

Serial. N0 0NHIa Z Z L

1 0 Heart

20 Chemical Pharmaco logy

3 ° C linical Phasmace logy

Be the sda? Maryland

ividnal Pro j ec t Report

Usico seric Efifie c t o f Flee in g

igator Br a J o J o Beans

meme

e Br o A lexandss B o Ge tmsn and Br o T ofi o Yfig Meants in0 New Yeah and flew flesh University Research Service ?\me sial Ho spital» New teak? Hes York e

dar yeae 1 958) Patient ways Qcalendar yearNone o

Te . inve stigeae the effec t cf Fleain and re lated

derivative on uric acid excse tien o

rial he nc e

gs Pienim (soasso lamine} has been widely need as a

sat fie s seveeal ye ssan Wh ile acessing the biochemical

in in the body it was toned that relat ive ly large

ccystelline compound was excreted in the urine of

had rece ived the dsego A lthough the crystals at

heeght to be s . ee caho lic presses of Flee ing they

_eric acid,0 and it was this ohsemsat ion that gave

we to the enacle se lsas nts potent urico seric propertie s ,

a considerably swee ten neiccsnsic effect than any o ther

ailable o 0s t sheervatiem has stimalsged ite clinical

eumste lcgists thsosghoat the country o it is expected

will be introdeced as a new dang fee the tseatmeat o f

seas inesre o

co the Exegese o f the Institate Studie s on hoe dsegs

acid excseeien increase en? general knewledge on the

y which varieas eateraliy occussiag ccmpoends are as

e kidney o

Serial N0 0 NHI— 225

orally administered dicumarol) obtained variable results o

A t pre sent , highly active soluble coumarin. anticoagulantsare available o In the p roposed studie s it is planned to

use sodium acenocoumarin (Sintrom) ,‘

e hich falls in this

category and can be injected at neutral pho It is hoped

that by repeating and extending Linkfls studies under more

reproducible conditions , information will be gained on the

effect of ascorbic acid on the response of guinea pigs to

counarin . anticoagulants o

The variability in the clinical response to coumarin

anticoagulants appears to be due in part to nutritional

factors o Besides ascorbic seidp Vitamin K is likely to playa role in such cases o Therefore s it is planned to study the

effect of exogenous Vitamin K on the response of guinea pigs

to acenocounarinq It is hoped that from these animal

experiments a lead may be .found which will indicate how to

carry out conclusive studies in. man on. the importanee of

nutritional status in coumarin. anticoagulant therapy; in other

words 9 on prothrombin synthesis e

Part B included

Serial No .3NHI=226

l . Lob e of Clinical Biochm i stry2°

3° Bethesda, Mao

Individual Project

of Exeee ineneal m anage 9 mDr e S o flees, Lahoaefie ay of Chm m Phaeeacoleg , m

Peoi’eesionalg

oxidase (nae) 9 W iel d. nee peeeene hhe eatahoiiea of parenterauy m m

isteeed sem teeino In eaiazele fife ia new ouideat that an altem te

m ate of m eeholisaaleading, to eem <tenine=0 e glacvaeonide effi cientlyaeaaholiz ee peaea

‘eem i eeeeeoain when aao is blocked, Serotonin -O m

glucnaonide was identified by enzymahic aehhede o

aete’

holiea of ingested sem tonin, Resum e? no eem tonin glncum nide

could he deteche in hm m im e Ia n e w eaeeeiaents with

Dan ssoeedsm g labelled eem‘aenia was M etered to patients who

had. been eeeeiviag m wfi é o A siege? m aahoiiee ape-feared. which m ine

to he ieeatifiedo In m eeaent aim aheae chaniani findings Dr, Leon

sem en-

rains in haein ie m ee dla in‘

hihihedo Following m e son-um of

3 fixmr‘

iZF‘

xl hee ia eeaeeevfla levels rise new fl uidly suggesting a possible

W ee am of ainnuee o W e w e es ea m anevee tie e e of: he ars in

pe einheeei eeee‘henin deneee o

it has also been shown ceaclueiueizr heat W yntezaiae is not con»

vented. to seaeteaia o Thi s was wh o wi th labelled tem taanine in living

Serial No . NHI=226

Individual Project ReportCalendax Year 1 938

Part B: Publications

l o Weiss'

baeh, flo g Reafieldg B ee‘s » and Uflemfi f’ienfig S o sunble

m s Peepeem es and A cfibiens en Sem wm oJ, Biol . 6mm , fi" 9536 9539 1 957 ,

2° Weiss‘

baeha e , m a ssed » D on. » M Usemfrieeaz, S . Binding of

Sesam e and seem » Amines by Elsea Flawless , A mmo Bieeewz, sea

N ew s . 1 953 0

30 WeiS SM 3 E 0 , Wa lkes, T o?” em waem iendg S o A Simplifi ed

me e fez? fleasee ieg Sem m eim m fiisseesg sm m us A ssay or

Both Sem m e and M em e , J, Emil. 0 chem gigs 865m , 1 958 0

1+ Fee-fiery Kc , Weissbsefisg E g g Reafi eldg B 0 6 .” Udeeffl eneg S

em u Am legees ef (samesew ) M W and.

as IeIsflbifiees si m e Bflesm fieesis w as N ew ef

Sm m o Jo Reese, Ghee, Sec o i933 3195580

Begdsmsm s D OE .” Wefilss‘

beeisg H o g seaWeeeffi eedy S . m a g ical

ew es? » Semflrem wphen Jo m o

and Exp m essage 1 223 Mac ias; 1 953

6a “dem ienfig S ag weiS SbQQihg H o g m aEm eg B oga A ssay M 88m m

and Remeed fifie’ee

'

eelieesg, h zymesg em me gs , “ w as ef c sm m

efieece Peblishers E g g 985 8 5 954 30 9

T o Udee f‘efiemd so see}. Weiss‘

eeeeg E 0 W ee? ef swmamw m eflse9

(Seem-

aw n in m ssees o Free s Se e o Exp M el a em . Zflfd o £ 3 71486 751 ,

80 WW S , T o?” Sjgleg’flm Q A 0 3 QW V ERM ‘Q 0 03 0 9 Weinsbachg mo p 8M .

Qfieefsfiendg S o nu n. mes s:pm m fiee and Re gee seaw eeds in

Bananas , Science fi g 6&8w65@pI11 958 0

w . sgeeeesee s A 0 9 Gw espieg Ego ” se eUdemfeiemag do A SW e fish es. for

we Be esem me ef B% m emime ©xidese W hififlee in men. Lancet 23

1 59» MSfio

Hessg S ag Weiss’

ieeehp H o g fiefifflelfla 15 0 13 0 9 esm Ufiemfzfiem g S o fi e

ReleefieeeM P Befiqeee Ipsefi ez m wesselfiez see {me M axim of its

Effect em m m (Om de se o Jm emo and M p g m essage 1 2193

1 2° Udenfi iemfip S o » Him , B o g Befifi aelda B OG”, and Weissbach, H o Studies

with Reversible Inhibifiees ea? new Oxide se o Hem lim and Related

m afi a Eieeme o M em eel o Ia leafle ss o

Serial do .NHL

1 . Lab . of Clinical Biochemistry

2 0

3o Bethesda, Mdo

Indiu m Project ReportCalender Year 1 958

m mu s s Biogenesis and sen seless of system s-on us and

and e , preamp, section of W esm ntel Therep omm

Eden Yess’ss

Total s

m afessionals Q083Others 2

Almom free mouse is a better precessor teen free bards-mgr»

pm line for we bound hydrom solflee , it he s been e stablished that in

rapidly grewing chick esfiovyosg es spem eieble as sent of hydrom roline

is directly ineoepors‘eed zla eon egeeous tissues ,

Preliminw y studies on we com esseoe of prelim so hydroxyc

prelim in e cell free system indieeee that flee product m y be a. coneJegeted foes of Hydrom m lioe o

Kemerou eeg em smeloé ee of meson-

imm u ne » causes a. prolonged

elevation of free mem e :[am ides in chick eebsyos end in the blood of

rat s . The im m en se for this elevedion has been established to be due

to (a) m on ies of mydsozggrpsoliee eseabelzlsrs by fizetoprolime and (b)

the laws); rescu es. is found in m e sepem e‘

ben‘

s fractions of m i: m ass};and liver and secuiees redmeed m im e m eleoeide o

W e the m elons m egs of pews-

eras stediedg only whose with

swim”em did nee

-

eels .“flee seized M me sedd exese

‘eioe by m ess

peeiemzs is9 however? memes else asse s]. sam e W e isom esed excretion

of mesom ue elime is em

"

; seduced by meeting see Easiest on a hydrowa

prelim fees dies fee two we eks 0

Serial

Publicafiions

1 0 mm , C . , sm m g T om , Friefi erg, F o g C ,

emblem of Eydm w m lime mm m em e m u m s by Chiek M amas 0Jo BM ) " Chem , In Press ,

2. film » 8mm , WIRE”, DeCosfieg R M. » Udsmm ieng Patchett ,A A . » and $11 t B . Studi es on hem e-Lew iee . Seiem e o In

30 mm » Cw Smith,”R E M DevifisemgJOB . » fidenfsiem a M oses ,

P age, 9 an d SJeesdsesg A . M m esasmes m gam es for mea nt-1m

marom m limes Applieem oe w Em Urine . J. Leb o w e Cline Efied ,

In Press ,

Serial No .“ 111 4 31

3. sem eada, ea.

Indivim m aect ReportCalendae Year

Progect“fig s (Sam uel A ssay of fin-Mew A drenaline 9 3m m

Eem flressllne and. 3exeetsewahafiydm zwse ndelic A cid,fi tehelites of A drenaline and Seredxenalm e

ass. Ms . E . marsh, Section srm em ew . Thee-3p .

wefessiesels 33(ashes

m aiden s eem have s weetly bees. seem ts be the eager m sebolites of

adrenaline and m m deeselsme in m s . wea sel essay foe shears estabolgte s

Ram se s in mesi ‘m em , disease as well as yield. infem tien en th e action

0 1? easy demise .

M em es adrenaline see. mem ememslm e have se en m u seumsecw eeed feast urine by eds eebmg en Ess ex Seweellmine , elmfiim mwe selm with mass ems. cem en t? essay sf the elsste with w ors e

m es. seem W e m ussels give mews es so soles sea as little as

e tag/em. sf see m eebelzmes «m mbe assem ines. som e seals m ce also

give little e? as eelee mu seum flee sewer high degree sf specifi cityslam med. u se the M ess: selfless . {mem e s fees; 6 patientsvies pheeem eym were 30 yes cent m sifi ve .

sight eeeem fi is. pew. fee was seselfss u se m e mes"melee .

mam sisg we arm s is essential so m esh es seslysis sf 3esteem serenelfime see. m eeeseesl fime Unfomm e

’eely been m ess am ends

presence sf Ess ex (W ) . W e seem se e a. high affi nity fer 3..m

Serial No .NHI~232

3. Bethesda, as.

Individual R e ject ReportCalendar Year

Part A ,

A cid and ywsmiacbatyric A cid

Qm er Investigawm David Abraham

Man Yearsz“Ecizala

m e ssiom l é

Others 1

“m e bicsynm esis of ye gmm idiacbatyric acid has been. shown to

occur in brain and resins of all species te sted and east act ively in

dog pancreas“

in additica to rat M assey }, as rapeseed last year . Due to

the carcass intere st in 7~ea.1acbatyric acid as a central synapt ic inhi

bim rg collaboration wi th Dr e E 0 Gm di‘

es‘t of Calm ia waiver-sit); was

initiated was the resale was ye m aidim basym c acid was found to

also be a patent cortical synaptic inhibitor , it is m fieresting thatthe me chanism of inhibieicm is di fferent free yo aaiacm '

syric The

paysiolegical siwificaace of flats seacticn is aalm cm . A ssay of

various eissaes Ice s revealed the widespread occurrence of ywm a1 61 m =

buWric acid wi th 10m in having higher was. average levels Injectionof yo geaaidinobuewic acid in rats was not accom m ied by any increase

in brain indi caiim flee im bility of 79 m idiacbufiyric acid to pass

the blood brain M icr o

M asaidimase is she saw cases catalysed the symm e zais of

yo gam idim batyric acid in W e 0 m e cad datiom and decaebcawla‘

tica

of m alaise is m e pafiasay in new im ef’

ce‘

cm tes amid W hey liver .

{This oxidaeioa getaway eswid m e be deacastcaeed 1 a the was" Teams

aei diaase can also caeaiyae”she sm eiiesis of fiem idim aleris acid

rm (5 asfiacva‘

iecic acid.» m is m « messingwas “me foam s: compoundhas cecea

'

ciy been ramm ed to occur in am en w ise

A new seam s has beea disccveced is iambic, liver and kidneywhich Wdrclyz es yanguaaidincbam ic acid fie fem re amincba

‘

tyi‘ic acid

in areao W e seam is ace aegiaase bee s ighs be identicei to"hececo

arginase"reported cases tie s ago to occur in: rabbit intestinal m cosa o

Serial No .NHL-233

3° Bewesda, Md,

23 211 171 3333. m gec'z

m geefit Tim es Biwmm esis and 3& 3m of ye GuaM dim mm ic A cid

and yeaM MW c A cid. 1 2 o Em m‘mm eM c A cid

M fessmm '3

3 12 1 3 332 M e acid is m m laye mfim mm ic acm e W e cm »

M 39 9 39 1mm? lam ina R s m am is m e

9332 £33 39 3 19W $9 3335 3339 913299 was $9 m m if brain

69 9. m m 5 929 7,c 9 9m 33 191 9 9 9 9 2’

(59 929 339 9 0 A siwia

09 339 9 3$m $9 is gm m mm Them m um m d $9 de$em fine if it 99 m »

m am 7°M 9b33m i9

m amm a c 959 39 was sym’é fiesflz ed 9m}, sm died £ 93: its vim

exaifl in $239 m

E2239 develewzem 9 2 a, M eal assay £9 3? 7m m ic acid has

wesem 199 5 9 93 923 its fi m efeed $9m (1 ) $39 fie$9m m 9m of

$39 333229 9 acid. m $en$ 9 3? QM» (a) $239 vim 9 1? m m m n ic

pm pem fies of W W M c 9 9 m . $13 a M ew 9 ? species? (3) we

w‘é mamcm 9 1? $29 299239 37293? 9 ? 23m m $9$339mi9 acid

in Lu m bfim sm g ( 13)“23239 de

‘ubemm mmm M

’

m’

fiaemi c ac id aetiviw33 3 333233 339 939m m qmmmg m m ami e (5) $239 bim ynm esis

9 3°

m m fivmifl‘eM 9 3 9m g 933231 (6) $239 92399339 3 1 9 9 9 9n gram mabum ic

acido

l . Kidney Electro lyt e Metabo lismZ .

3 . Bethe sda , Md .

PHS o NIH


Calendar Year 1 9 58

PART A

Pro j ect T it le Effect at Strnpnantnidin , adrenal stero ids , and

Vasopressin an e ster and e lectro lyte excre t ion

in the calcken .

Princ ipal lnve st igate r Jack Orlctfi

Other Inve st igatcr'

manric e darn

Man Years Pat ient Days NoneTo tal

Pro fe ss ional:

Other 0

Progre ss

The chicken was enc sen far the se stndie s finr reasnns ontlined in

de tail in the last progre ss renert . the renalo

ncrtal c ircnlat isn. neke s

it po s sible to invest igate direct tnbnlar e ffect s at narisns agents byinj ec t ing the

_te ss snbstsnce late a leg ve in and cannaring nrine detained

separate ly firsnaeacn kidney Changes in ccnnc slt isn cf nrine Eran the

inj ected side are always due to tabular ellec ts .

In tne prev ious renert it and been acted that the cardiac aglycone ,

strspnanthidin inhibited the transpnrt system in tnbnle ce lls by which

sodrwn is reabsorbed in exchange rnr pctass inn. and hydrogen inns . In=

j ec t isn or the drag nnifsrnly re sulted in an increase in water and sodium

ercret icn and a tall in nrine hydrngen inn cancentrat inn . Changes in

ps tass inn excret ion varied with the erperine ntal ccndlt isns . Strophan

o

tnidin decreased pctassinaxer

cre t inn if init ially high and enhanced or

did not street it ifi init ially lee . The re sults are cans istent with the

hypothe s is regarding the site at act isn c f the drag s ince the slight inc

crease in potass inn encret inn nbserved when encre tlcn was lea init iallyis pre sumably due tn the de livery at nsre scdinn to the exchange s ite

and re sultant increased exchange de sp ite an sverall redne t icn in the

capac ity o f the agatan .

Potass ium is kncnn to interrere with the act ion o f strspnanthidin

on e lectrolyt e transpnrt in red ce lls . The interpre tat ion o f this effec t

is unc lear . Bswever , enn ilar re sult s have been noted in the ch icken in

that strspnantnidin induced natrinre s is is diminished if large ancunt s

o f potass ium are inj ected s inn ltanennsly .

Part B inc luded Ye s .

Se rial No .

“ 1 4 31 5

D irec t ion o f current re senrch

Strophsnthidin end s ldo sterone s tre e t s rennl co rt iso l s lice s

rebbit s ( end dogs ) on e lectro lyte transport ore be ing examined to

prov ide intorne t ion on the prec ise node o i ac t ion o f the se compounds .

Se e s ssoc is ted progre ss report .

inc idental findings

The e ffec t o f KCl on urine concentrat ion was inve st ige ted in the

chichen s ince prev iou s so rh in the dog in this lsho rntory indic sted

thot dilut ion nsy be interfered with by sdninistrst ion o f not e s s iun

ss lt s . This us e no t continued in the chicken de sp ite inj ec t ion of Xcl

into the renal portal venous c irculat ion .

Sam e ], No NHI“ 235

1 , Kidney Electro lyte Me tabe lism

2 .

3 . Bethesda , Md .

PHS NEH

Indiv idmel Pro j ec t Repe ra

Calendar Year 1 958

t: T iazle Efifiece o i cardiac glycaem ee end adram l stero ids

on m fiebe liem 0 5 slice s of renal cexmex

pm Invest igator Jack Orlefii

Inve sgigetor Maurice Bum-

g

Pamiem Deye Nome

Deecxigeien

Effect: of eez dim: glyee e ide em ! adremel seem ids on

m aebo lism , ez ygem cemeempeioa , (zed pe w- amine »

eke oi kidmey emce e .

Qf rabbit; t eam ]. cent-tea were prepaged by m e techniques e f

animals were exem gmimeged and the exc ised kideeye we re

. 3 w 0 S m slices e f metmal w rgezz were im anbet ed at 2590

wages: e w lmtflm flex em ice mm hem e ,

im am 9 ? M m]. e fi m e cez dm e eglycm e g werephemem dm

a M e et ing efi slice pegeeeium cm gem and a propem mmel

slice sodium ew eemt . m e mpeehe efi eme w e use M um was

Oxygezm m emapm em wee cm affiecged o

ipm ee l chemge e m cetgimn cem ent; ere pz eem bly daze ta

m m e exam-18mm mechamiem which mama up Kfi' in ezceheege

Bie'a'

. Studies gapem fisely cemfi m thee we m fl m :

ee lly effeceed .

e in PM eecm legim is e im ez due rm a dime“ effece

he firem mmt (M am eysmem em is me eezmdery to the eew cieged

aiam free we m em e . A 9; flew slice pe fie ee im eemeemtretmms

ekem up , even be the m eem e e fi {the dm g. In experimem s m

fl ea-33 1mm was m aim ed by imeubaaiem m we pege eemm medium

Serial No .MRI- 236

l o Kidney Electro lyte Metebo liam

2 0

3 . Belhe ede , Md .

E33 0 NIH

Individual Pro j ece Repe rt

Calender leer 1 958

'

@ j ece T iele Kie eelce e f Sedfiee med Pomeeebwm exchange in

f eeble kide ey slice s

le c ipel leve eelgemer deck Orle fif

chef leveeeigece r‘

Mee e lce Bemg

se leere Pelleme Deye Nee e

leme l Will

Peefie ee ieeel: 7/ l2oeeee 0

j ecelve e

e lep e meehed fee eeee eeee deeermde emiee efi eedfimm and

filee e e in klde ey ellc e e end epply eels me e eeedy d f lactat e

efifiecm meee l electre lyee eeee eperg .

ere ee dexieg gee pe ee yee e

hie eeie e e l We?end K? eeeeepeee in klde ey slice d be e heee

by eeepexleg tee er le es o i the spec ific

ualme e dififiexeml eee efi e llee e le e e ece e f gee

Se ek experieee ee leek pmec ieiee e imce geese megeeee eee indiv idue l elice e eed eeverel eee lyeee ere

e deeeme leeelee , ee lelplyleg peeeible verieclem due

Alee , lmeerpeeeeeiee le cemelieeeed by flee face

leeeepe cemceeexemiem ed the medidea ie cemeeeeely emeegieg.

lee eeehed d eed lm eke eee eeee eemdlee e siegle slice e fi rebbie

reee le perfe eed wi th e eedlemxef eee eeeee leeeepe eee cee cz et lee o

e le kepe la a fixed ve lwee ef able mediemaedehle e we ll

melee cemeter o Simee eke ee lmee end ieeeepe eeeeeee e fi @&e

l ere eee eeeee , cheege e ie eeemeleg gate ere due

leeeepe game the elice . The eeme ellce cee be

he ll eeeeee l amd eepee leeeeal deeermie etiee e .

llmdmeey eed die e eheeed ehee ellce e perfe eed ie ghee eeee er

l eeygee eeed Keebe o llegee elce z eeeeee ee le elee ea 25®C ee imee im

ee ee 2 to 3 hemee e eedlme . emd peeeeefiwe . c@e@eee smaller fie

fire ehly exc ised eleee e .

Ser ial Ne oNHI~ 238

1 . Kiduey Elec tro lyte Me tabo lism

2 °

3 . Bethe sda , Md .

PBS NEH

Indiv idual Pre j ect Report

Celeedet Year 1 95&

t T itle : fict ive t+ flee fie t eeemetttmted tween erythrocyte

ghe ete eed its t eletiee to me tebe lime

ge l Keve et igeter Joseph F . Hetfe ee

have ettgeter Demie l C . toste eee

Petiee t Days meme

t De ee t igt ieeo

t Perpe ee at te eeet et

it e seaside teble bedy at ev idence that the active transport

ec t e ee the hemem t ed bleed ce ll is eeepled to it s me tmbo lism o

ted et the teee tifiieet tem ef the pet t tee le t t eecttem ( e) te a

tty at the me tebe liemt pt e eee te

e t eeet iee . The eppre ech at

ten , e e ieg t eeee t t ituted t ed

tee leted ead assayed fer

e t the ghe t t pt epet et iee

t ttet teeee , me tme lly , eee o pe tme et teg ceeeee ee te

te te the gte et te te t tet , dat ing the t ime e t heme lye is ,

itut tee e f ite ie it fiel Kt perme ability etet eetero

certain phye iee l emd chee iee t ptepet t ie e e t the

eme rged ehteh'

hee e beeewe te lpfe l fie wedereteedimgettmetuz e te tet et tem.

the pe st yeat

e t ea ee ttt e eee t eptedmc itle gte et system , it tee

t etemt iem eepee ity eee the Re filex it se lf at me »

ia teeee t ete t e é epeedee t . This is ille ett eted by

em teeetet tee e t 3?@c te ctee ee e the Re t etee t tee

time eeee aflo te ld . Thte recevety firemxheme lye ie

ee eeeee e ett e e tut e te t epe eee ee the pass ive

eterectee ietfie e t the e t tgie e l pepmlet tee epplie e to

t the de t ivefl ghe ete , Et tee ae t be en pe eeitle to rese t

eetiee e lttet gt thte tt ect tew eem be diminished in s iz e

demeity tepe tetmee . teeee et ite ted gee ste new be etMMo

late K+ ege iee t em fie fe le gt edtemt by the eddit ioe ta the

eme lee eidee . the fie that at eemt t e l gte ete (ea added

efitee ted by ettephemtbidtm; em the presenc e e fi tee e ie e ,

gaze t te-at he .M il n e}?

1 . Kidney Electro lyte Me tabo lism

2 .

3 . Be the sda , Md .

PBS o NIH

Individual Pro j ec t Report

Calender Veer 1 958

text h .

Pro j ec t Title Alkali e etieh me t ee eet it heh eh erytht eeyt ee

eftet h ltt ewie le t fit t ediettee

Pt te e ipel Inve stigate t s Jehe S . Geet (Sh e et We tter)Jo seph F . hetteeh

Other leveet igete t meme

Pet teh t Deye Nome

Gee et e t P erpe ee e t teeee teh

ted se tle expe eed to h it t eWie le t light eede tge e ee lle id e eme tte

swe lling which leads ev ee tt e lly to hee e fiyete . tree prev iews t ext it

we e fet ed thet the t ete at thte hee e lye te t h e ptepe t t tee e l te the

equet e e t the te te l ulttev ie le t de t e . The pe rpe ee e t this we ek is to

cheree te t iee the nature of the rediet iee le e tee ie terms at memht eh e

permeehiltt ie e e e eh approach to eh det etehdieg the de ee o tqe ered

t e let iee ehip .

trott e ee tremaj eh e 1 958 te Seeteehe t idhh

The prth e iple ehee tvet iee s on the eevee eh te e t he and K at t e st

the fit t edie ted t ed ce ll membtd ee ewe e s te llee e : ( 1 ) th tt eee lle le t K

e eeeee e eeee tdihg te first e tde t hih ettce eed that (2) the me te eee stee t

for K et t tldt is pt epe t tieee l to the square of the te tel de ee o f ultre e

v te le t light ever the tit e c te ld t ehee te sted . ( 3) the K te tlex ie

prepe rtieh e t te the eeh ceh tt et tee h t K in the me dium (th e e ll de h es) .

(h) the t et ie at the the e td to eh twerd z ete cemeteh te te r K , at ditt=

erde t de ee e , te eqe e l te the ehte t ide t et ie t .e . ( 5 ) Pee e ll do ses

te sted he flatten t h e fet ed te he pt epe t ttee el te the eeecee t t e t ieh of

he in the ee te tteh e t e t igte t ed that (d) the t et te @ t the t ete

cemeteet e e t he th tle t te K outtlt t te teh ed te he eee etee t ted equal

te the ret ie fer peee tve pet e eehiltty h t t h it redteted ce lls t . e .

Coh e lh e teh

Ultrav ie let light greatly tee t eeeee the tt of he th K ted he

at t e st the eemht eee e t the t ed bleed ce ll . The ce ll heheve e d e though

Part B inc luded : No .

Ser ial Em N H i- “ 4 LN

3 o Bethe sda , Md .

Pee o Mid

Individue l Pro j ece Report

Calendar Year 1 958

Fast A .

Pro j ect T itle Ceeiem tz ee eeere fie high end lee peee eefiwm sheep

red cells

Priee ipel Keve eefigeeere Je eeeh fl .

Demie l C ° Ee eee eee

Oeher Imveeeigeeer Nome

Peeieee Days Nome

Gee erel Purpose of Re eeeeee

Seme indiv idual sheep eeve red ce lle wieh high peeee eiem. eed low

eedieea (HK type ) eeme eeereeieme while eehem sheep heve red e e lie wieh

high eedie e . eed lee pe ee eemme (LK eype } eee eeeereeiem . Keceme ev idence

o f Evens sugge eee ghee the EK chereeeee is inherited ea e'

Meede liee

demie eee . The ee eee ee of twee reeeeech fie ee de eeemie e experimeeee lly

ehe me med K peemeebfilieie e of eee ee ewe type s e f she ep eed ee ee ee

queeeieemive ly e derived fierme l descripeiee e fi eee ee membeee e prepe t t ie e ,

the ee cee e efie l epplieeeiee e f eeie eeeery lieke eegeeher the membrane

eeexeeeeeieeie e meee eeery fee the ee ieeemee ce e e fi ewe diffie eee ce e fie

concemereeiee e ed Ne med K ie ‘ eeeh eype of red ce ll end prev ide e an eg o

plee eeme fer the meehee ie e fi e eemrege leeiee fie iediv ide e l ce lls .

dregEe ee derieg eke page yeee

.e deee iled ceeperieee efi gee K end He eeee epeme prece sses fie H R

and LR sheep red e e lle bee beee'

eede . Meeeuremee te dee ige ed ed separate

eceive tremepeee, exeheege diifie e iee eed peeeive dififie eiee eeeeee ee ee e f

K end Ne flee e e by eeehe iqe e e ievenv img vee ieeiee e fie eke eeepe eieied o f

ewe medidexeed eke wee e fi diffemeee embeeeeee e end etrepeee eeidie permie

eke fe llee fieg eeee le eiee e : (l) em eeeive eree epe z e seepeeeee e fi K imfilwx

secure ia LK es e ehl es HK e e lle but is gee r emme e eeeeeee fie eke letter

type . (2) en eeeive eree eeexe eempee ee e e d Ne euefilex be e beem ident ified

in been ce ll eype e . (3) e large feeeeiee of fine meee l Ne flex e cemre byexchange diffe eiee in me te ex awd LK ce lls . ( é ) gee pee eive perme ebilieyto Na amd K of em med BK e ekle diffiee me rkedly . BK c ells have e greeeer

pess ive pe z ee ebiliey te K emd e eme ller pe eeflve peeme ebilfley fie We then

Pet e B ie e luded

N i 1 Ni} "

w m m

kfifik gel

m w m w

71“

l . Mide ey E lee emd lyle

Me tabo lism

3 . Beebe sde , Md .

PBS NIH

Indiv idual Pro j ecm Repexe

Celeedex Year l958

Pee j eee Elele Bedeemieeelee e d the Tee e Chloride Cemeene of T issue s

Pelme lpel leve eelgeeee Eee e ee Sealeve

oeham leve eelgeeee Meme

Peeieee Deye Rene

fixe jeee Be eeelgtlemo

demerel ee eee ee e d fleeeeeee

Chlee lde is keee e fie be flee predeedeeme ee lee eeeeee e fie aware »

ce llulaz Ele ld , be e lt d eemeemfireeiee la ce ll fluid has beem very

e eeerme ie . Re lleble le lereeeiee em degreee lle lee cele eide dee ld be

impereeee in medereeeedleg eke beeeelee efi e leeeee lyeee le eke body .

The dlfifilee lelee fie de eerede leg degree e lld lee eele e lde have been twe e

fie ld : ( 1 ) eke leek e fi e K elleble e e leexiem fe z eve le efileg the verlee s

emelyeie ee eeeee le e te ee l eisse e ehle e lde emdee eeve yie lded wide lydlepeeeee t e de les ; emd (2) the leek ed e reliable de eeed fer e edflmeeiegexeeeee lle lee ve leme , end eeeeeee gee fieeeelee e e f eeeel eieee e ehle ride

which ere eefireee lle lee eed leeeeee lle lee

Pregee ee Dd rfieg ehe Pest leer

The fileee e f gee preblee e defied ebewe hes beem de lved by epplieeelee

of eke prime iple e d ieeeepe dile elee , leve lv ieg eeepleee mde ime e f t is sue

eelexide week medleeeelve eeleelde o le , ead chemical lee leeiem e fi ehle ride

fie ee eeeeelve eeegee efi pmelfiieeelee fie eee eeeee epec ifle e eeiv lty . This

ereeede re be e peev lded em ebee le ee eeemdeed ed eeleeee e e fer eve le e eiee

e fi elep lifiled me ehede ,

The Badge s in eke leeeepe dlle elee . e eehed ere : (l) e eemple o f

fresh or de fied ele ee e le eempleeely dige eeed le beg dile ee e lke li e leh

added Chle elde wad prevldleg eeeeleee edg ing 1m ee le eled e f eeeble end

xedle eceive celemlde ; (2) e peeelee of the e leeli ee ld elem fie dried ,

d ebed de high eempeeeeure , and eke emgee le e fxe e xe elde e redisso lved ;

(3) e part ied e f ehe e efi ee lmmiee le teeeeed e lem ee ld permeegeeeee in

e werbe rgo eype Eleek , ee idle ieg ehleelde me ehle ele e ge e weleh dififie ee e

de ed eke e ee eeE ae e ll ee le elee efi very dlle ee e lke lle e hydregee perexlde

edd is reduced ee ehle e lde (exldeeiee w z edecelem d fi (G o a) ee lemiem) ;

Pare B lmc lwded

Ser ial No NHI'QM

3 . Be thesda. Md .


Calendar Year 1 958

Pro j ect T it le : study ct the em ceh tre t ihg end dilutim proce sse s

ih the m im ic hidh ey

M im ipe l Ihve st igetor Jehh R .Jeeh iheOther Imve et igetcre Gecrge A s Bray ,Jt .

R . w. Berlih ct

Tote ].

Pre face ieh el

Obj ect ivee

A h e lehere t ieh cat the mecheh ieme epere t ive ih the ceh ceh tt et ioh

sued dile t ieh e t the crime .

Preht e cc De c-ice the heat”fleet : (Started w rit N IL/ Sh)

The me j er emphasis he s be en eh sett ihg hp h e thcde fer the evaluat ion

of chehge s eccmrt ihg ire the distal t elne t tubular eyetem end in the ih te

st it ie l flu id o f the hidh ey it} teeter dimrecic eh d it ch t idicre sis . These

hhve ih cleaded

( It) A m dit’

icet teh c f the cehweh t ie h el etep e flmfl eh elyeic which

large ly e lie ih e te s the pelv ic dead space eh d emce e collec t ion of

smell em le s which direct ly reflect e lte t et ieh e 11 h c rime cm e e it icn

de rives; the pet-ted c t etepped flee .

(2) The wee c t eediomh ceh e it ive e lectrede e , deve leped hy Dc . MurrayEdema, tc mee eh re directly chchgee 11 h sediem ceh ceh tret ice withic the

rem ]. peceh chyh e dc t imggm aim ed chm e e am free water c leereh ce ih

the deg.

(3) The deve leph eh t , hgy Dr . Gee tge IBM }; in thie Eeheretccy , o f

teche iqme e tcr tedfie a cc tegt ehhy m hidh ey acet ich e

Evelcmt ieh c t e mic te e methed tee sediem determthet ieh ,which

hee been dev ised by Dc . Rehert 13m m .

Part B inc luded

Serial No ,NHI « 2t5

3. Be the sda , Md

Individual Proj ect Report

Calendar Year 1 953

Part A .

Pro j ect Tit le Studies as Diuret ics

Pries iesl Imee st ieeter : T . J . Kennedy ,Jr .

Other leveet iester heme

hee‘

tesrs Pst ieet Bees : 279

Oee e Third ( ll3)Pretessieesl: Oee e Ehird ( l/ 3)Other heme

Ohj ectives

The geeerel ides s t this type s t stedy is te e t ilise the street of

eiereties te eluc idate the essie eechse ises ieve lved is the reesl tree s e

pe rt e t e leetre lytes .

Pesere t e se ries the eest yesr

Steeies eeeertshee is 1 957 eh the die retic streets at

chlerethiss iee were ceeeletee. Eih e i seecle siee s were lereely these

set ic iested ie the prev iee s yeers resert . Chlere thieside is sh

e ttect ive ers l die t etic ih these meetseees e t eeeee ih which sslt

te lersece is est tee heely thee iree se t ih which severe seceeesry

hyperelee stereh iseneees eet exist . while set es eeteet es mercurial

die setics , it he s edesetsges ih terse e t esse e t eeehe istrst iee , etc °

thet mete it s vsle ehle therseee tic egeet ih c lie icsl eiteet ieh s

cherscterisee he seems . ih these ee re severely ill est ieet s whose

se lt te lersece evee st hed rest is were lee she is t hee severe seceedsryhypere lee steree iee resists , the ereg is s seslly ei thee t sige ificset effect

em sediee excret iee eeee ih ie steeces 1where e erce riel diuret ics sloee

er with W h it e chieries ere effect ive , Ottem, ih e ach eetiee ts ,

erhihit ieh . et chlerethiss ide rese lts ih the ieee ct ieh efi eetesstem

deplet ieh with eccerreece st hyeehsleeic slhsle sis .

hireetiee e t Cerreet heseersh

etediee eh chlerethies iee es s die t et ic ere esseet ielly ceeeleted °

lec ieeetel tiheiegs et sigeitieeeee

ethelee eh eee efi several est iee ts is this series ehs eresested

smth hyeeeetreeis 9 hyperheleeie ems scieesis were else cessleted ° The

Part fl inclesed

Serial No .NHI~ 2A8

Experimental CardiovascularDisease

3 . Be thesda , Md .

res are

Individual Pre j eer fiepore

Calendar Year 1 958

Pe rk A .

Pro j ect T itle : fihe Phye ie legy efi Gomge erive Heerfi re ilmre

Prime ipel Inve eeigerer Jame s 0 . Davie

Greer leve erigerer fi iche le e A . Yemkepeule s

Ewe Yeere Periemr Days : Nee e

feral 2

rre fe esieee lz 2

Other Q

Pro j ecr DeecripEiee

Obj eerfive e

A rea E . Te derermmme the me ekee iem. e fi imeree eed e lde ereree e

secreriee fie eeceedery hypere lde ereree ime .

A ree 1 1 . Te define ewe bie ehee fieel de fecr fie eke reeling

weeerdiem .

Pregre ee Durieg rhe Yeer

A ree I . Mechee iem er elde erereme Secret ion m Secemdery xiypere

elde eteree iee .

Pre j eee I

la . riele Eeeree eed A lde eeeree e $e ereeiem Fe lleeimg; é eere Cee erriegien

er the Ee fierier Veee Cave .

1b . Eereerigerere : Jeee e 0 ° Devie , rereerd Kliwme g Niebe lee A m

Yamkepemle e eed Relph E . Pagereee were eke surgical e ee iecemee e f A lfred

Ceeeer .

Pregre ee Be rfieg the Yeer : Pre j ece ia cemplere eed paper bee been

eecepeed rer pe blieerfiee im geeJemrmel er Glie ficel Keve erigeeiee .

Eee erfieeee er eeere eee eerierflee er gee rherec ic rererier veee cee e

( 7 dege) end er eke ebdeemme l imierier veee cave ebeve eke edremels (4 dega)em e lde eeeree e , certice eeereee eee Eerrero efilber erere id see reeiem 1 m

edree el veia pleeme were eemdied . reree er the dege wirh ehoree ic ceve l

cee errieriee rece ived derrree recreveeee ely re me iere im or to re cree ee

ple ee e we leme . array 3 seerrel deeereee erree e 30 min , eperfi , eee eure e

mee ts were made er emmiler re eerwele dwrieg eke experimeere l period .

Fe llee ieg rheree ie cewai eee errieriee e lee e , e lde emereme secreriee inc

cree sed withfie 30 min . ema reached leve ls fie ld greerer rhea the

average eemgre l rates or ce rt ice erereue and

Parr B iae iuded Ye s .

Ser is l Ne .

fialm zha

0 3 0

The e ffects of hypothslenic le sions were studied in 1 0 dogs with

chronic esc ites produced by csnstrict ion of the thorac ic interior vans

csvs . Bering the first 3 postopers tive dsys , the high rste s f s rinsry

sldnstersne excret ion end the nerted‘

ds retent isn ensrset erist ic o f

deer wi th enrsnic experimentsl sscite s were eneltered in all 1 0 sntnsls .

Every eres of the typetnslenns ens destrsyed bilsterslly in at least one

sninsl . 7 of the no dogs surv ived the snbscnte 3 day period ; 5 emine le

lived fer 2 nest s or longer but 2 dens died stter d dsys . In the div e

ctrenic enteeis , e ldesterene encret isn in. mrine red e fined e levated stove

nsrnsl sltnengn in 3 entnsls s prsgre ssive dec line secsrred; ssdisn

escret inn rene ined lee in ell degs snrv iring the snbscs te period . In

the 3 sntnsls in which sldssterene detes t dec lined end in these dogs

snly , there s e e bilsters l injnry s t tne. nedien esdnencs end sdrensl

streets. Tee dste shes , theretsre 9 test the ctrenic type tts lsndc

les ions were without street unless the nedisn eminence e ns injured .

tre j ect IV

é s . T itle : Evidence test s dennrsl eeent St innlste s the Adrensl Ge rter

ts secrete eldestersne in Esserinentsl seesndsry Hyperslds stersnisn .

4h . lnvest igntsrs : Nicns lss d , i sntepenle s , lane s 0 . Denis , Bernerd

Klinsn end Re ign E . Petersen.

ic e stat er tireless is s ie ves w ire less »

Te test the possibility st s‘

nsnsrel eiterent ne ctsnien tn the

regs lstisn. e i s ldsstersne secret isn , cre ss c ircnlstinn experiments were

pertsnned betwe en dogs with ttsrsc ic interior s ens csvs csnstrict isn end

nsrnsl recipient sninels . tees with tnsrsc ic csvsl csnstrict ion secre te

lerge qnsntit ies or s lde stersne end sens eldest csnelete Ne retent ion .

Cre ss c ircnls tisn ens esteblisned ttrsngt the rensrs l vessels , er the

ins isted sdrensls s t s nsrns l sntnsl were pertnsed by the technique of

Hiltsn, st s l . (dee r .J. Ptys ie l . l922525 , ltfid) with bleed tron s

hyperslde stersnenic dnnsr . central end recerery perinde were obts ined

sets the rec ipient's bleed er by cre ss c ircnlet isn or blend from s

nsrnsl deg. Resested deterndnst isns s t sldssterene end csrt ics stersne

we re ends in sdrens l re in pisses by the rsdie ise tspe deris st ive ne tted

(ted . Pres . l7:255 B l958) . t lde sterene secret isn in the rec ipient inn

cres sed il@% (7 deer) dnring c ircnlstien er bleed Ewen dsgs wi thsecsndsry typersldsstersnisn end retnrned ts tee centre i leve l in 3

nemesis in snicn e reces ery neried ens snte ined . Cert iesstersne

secretien incressed slight ly ( tea sers in 4 s t 6 degs . Ne consistent

slterstiens in pisse s ssdinn end ss tessinn were detected . Venous

pressnre in the rec ipient dsgs ens nnctsnged . Bering crsss c irculat ion

s t bleed iresxnsrns l dngs ints tee iss lsted sdrensls s t nsmnsl es is sie ,

slde sterene secret ien nee unchanged e r decreased . It is resc inded tes t

s nnnsrsl egest st innlete s tne sdrensls ts secrete s lde stersne in dsgs

with secsndsry hyperslde stersnisn .

Pro j ect r

5e . T itle te letisnsnip sf ddrenncsrticsl end dnterisr Pitu itaryFunct isn to the Eecs l Escret ien s t sedinnasnd Petsss inn .

Sb . Inve st igstsrs : Jess e 0 . weris , Wilnnt C . Be ll, Robert C . Bats , and

M; s Gssdt ind with the snrgicsl e ssistsnce s t t itt ed Cesser .

Seem l do SIM - 248

es 0 . end Wimeee C . Bell ,Jr Effiecae of a body ease

reee eed eediem. eeeeeeide ie dogs with eepeeieeeeal

em.J. Physiol . , 1 92z538 9 l95d .

8 , Niche le s A m ,Jemes 0 . Devie , Ratearé mlieee end Ralph

e : Ieeeee eed e ide eaeeeee eeceeeeee fe lleedeg, ece te

we efi eke z ecie eed ed ebdeedee i ie fee iem veea cave .

i7zl73 , 1 958 .

es Rebeee C . Bebe , Nieee ie e e m ieekepeeloe , Bernard

ape E . eeeeeeee eed Wimeee 0 . Bell , Efifieeee of eee ee

e eyeeeee ieeie iee iee e ee e ide eeeeee e eeeee tiee in dege

ie experieeeee i e ee ize e . flee deyeme leeiee 1958 .

e , Niche le s A , eedJeme s 0 . Devie : Geeee ic eeeee etive

deee wiee eeiee epid ie ee ifie ieeey eed pe leee ie sceee eis o

Leeiee 1 953 .

ee Miie ee c o Ben ,Jr . eeee ee 0 . Been eed ii.JayRe ieeiee eeie e d eeeeeie t piee ieeey eed e dreee u ee z eice i

e gee feee i eeeeeeiee e e f eedie e eed peeeeeeee . ‘ee .Joo (A eeeeeed fle e pebiieeeiee ) o

ee Eeeeeee Kleeee o Nieee ie e e , i eeeeeee ims eed Ralph

Ieeeee eed e ide eeemeee eeeeeeiee fe lieedeg eee ee

efi flee ie fiexier veee eeve o Jo clie o iee eee .

Set h ]. em,Nail

E, Clinic ef surge ry

3. Beche ede

PES 0 did

dedivideel Ere j eee Reeeeeceleeder leer 1 958

Eme j ece lielefi Leia Hee z e ceeeeeee ieeeien

Feieeieel leveeeieeeem: Andrew Ge meeeeeb ea Do

Other leveeeigeeez e Ketelle Geeee

Fred Belleek‘

dllliee . Leege1 in

men leeks (eeleedee yeez 1 958} Fefiieee Deye {calendereeeei : yeer 1 953)Peefieeeieeel:

Oeeex fillz

Pee j ece meeeeipeiee

eeee eee e peeviee e eeeeeee heee deelfi wieh ehe eeeeiiqee

eyeleeeeiee efi leie eeeee eeeeeeeeieeeiee in gee e fee eeeene

teee ebeeeehiel lefe eeeee eeeeeeeeieeeieee'

eeee been pe z feeeed

m em e e deem e a eexiee e eedeel. h ere h e eeee im eee eiegeyelieeeiee ed leie heeee eeeeeeeeieeeiee de gee seedy ef peeieete

eeeeie

end teem“ m eeem ee e by

ie eeeegeeeeea indieeeex

dile eiee eexvee gwiee. le£ e eeere ie j eeeiee . eee elee being eexeied

Gehee eeeeede efi le fe eeeee eeeeeeeedeeeiee wee eleo beingweedo Peeee eeeeeee

-

eeeeeeee ef eee left veneeicle has been

eeeeied wee in neezly lfifi peeieeee o ibis eeeenide e i s 0 5 pereie

e ler epplicebiliey in yeeeg childree end in eeeee peeiee te ween

ieee eke lefe veneeiele beeeeee eeepieellya Xe eeuy ineeeeeee e

ceebieed eeeeebeeeeeiel eed perce eeeeee e eppge ech be e been used“

seediee eeelie ed ebeve exe

peeeeeee ef gee lefe eeeiee ex lefie veeeeiele eave beee . cet eied

ee e in e lieieed. eeemee efi peeieeee o fieie eeeeeieee eeee ed eeeee

eeeeeee in gee eeebeeeiee of edeeel ieeefidieieee a flee le eelieeeiee

e fi eeeeic eeeeeeieg end gee deeeeeiee efi leeev teweieee eeenee a

Eeee B ieele ded de e

Seriel ed .RBI- 250

1 . Clinic of Surgery

3° Beeheede

PBS NEE

Individeel Pre j eee RepexaGeleedex ieex 1 958

gee Eitle sevelegeene end Clie iee l Applice tien of e fibdified

Eeye cee ee exmifieie l eeeet end Leeg eeceine

eipe l leveeeigeeer: dedrew G , eeeeee b ml Do

5 Eeveeeige tee e eeeeee'

e, Gilberts ml D z

deem e a Weldeee eee , id. D o

Rebeee e; L. LeegD ea Do

Clereeee So weldeea ML D o

deem Re esa Ed, B.

Eeeelle Cehee

Behexe Cere

fired Be lle ek

flee t s (eeleedet Eeeieee Eeya ( calendaegel 4 year 1 958)efeeeidee iz 2

2

ace weeee ipeiee

A . pz evieee eeeeee ee e deeeiled eepexieeeee with the

wee eeeifieiel eeeee eed leeg ee chie e‘

eeelueeed in this unit e

eeeeee e wee ebeedeeed fee gee fellewieg xee eee e : tee

eeee ee ef eeeeeleie ed. eeeeie e bleeding in 5 pe tieeee ; ieebilieyme deveee ee eeygeeeee leege eeleee e ed bleed edeqee tely;

edeeee epee eeeeieel eeemildeeeiee ; e high ie eidee ee of e ir

Ee Febeeeey l95d e Reye deeae eeygeeeeee e ee perceeeed°

we e ieeegmeeed ieee e . eeee end eeeeeel unit deeigeed end

a eeee. eee seepeee eiee ef the eie Eeeeeeeeee Seceiee o ihie

le e 8 in iee eeeeeee fieeee eeeeeee e fe ll eeygeeeeiee ef bleed

lee e efi 4 i .fimie a eee geeeeeea‘

eey be eeeeleeely deerilieed

eeeee ie eee de j eeiee e fie bleeda end deed meg predece

ieieee eeieeeeee ee meee le in ede embelieee fee eeeeiee he e

epplied. ie eeeeeeee eebj eeeed ee eeee eeereeieee fee the

eceiee efi eee eee e eeegeeieel eee eeeeieed ceedieve eculee

W G o l

Serial Rb .NHI° Z 51

1 ° Clinic of Surgery3° Bethe sda

PES e NEE

Individual Project ReportCalendfir fiaar 1 958

Prgj asa Eigle : fine Dimgnamis of Eulmvnic and Tricuspid

valvulax by Eye -Dilution

Emancipal Emveafifigafiar : m; Pemrymmn Collimao fiL D.

03m g Enve eaigfiam s Eugene Bm mmvaldg E , D .

Andrew G o {3&3v m, we

fiam fié firs ( calendar yevm 1 958) Pamiena Dfiys ( calendarfibsml é year 1 958)Pxofie sai@aalz

35

Prvj ecv fiaaexipgivn

Patiemga‘

vizb auspected imcvmpeeence~vf the pmlmmnic and]at fiz icuvpfid vakvea hvve been sawdied by mfi vma vfi the inj ecaianvi the fimdicagvr dyes cardfivgveenb intQ gha disgal chamber

while blwvd as samplad cvmtfinuvvvly ahxwmgh a densitameter fram

gha pt©8imfl 1 chambeg o En ahfim mannav .amy dye which t egurgitatea

acravs the valve inav the prvximal chamber cam be dete ctadodavble lumen caameger was cvnstvugted ia amvh a. mfimner ea that

the lamina efi ghe sepaxaae cvghegers s z e 5 awe apvrt . Eighg

nwmmal valvag have been ammdied and fig has baea £ @und that am

regumgitafifivn sccuvs frmm. ghe caahe ter 'a having bven inverted

thzvmgh tag valvao fihevefiwvea fihe appearmnee vi dye in fihe

v xfimal chamber gxaafi emvugh n duva fiafinite curve is

divgmvvgia vf valvular regvvgigativn.

g'

wH M g ._ q Ovatinved. use as a standard

aechnfiqme fur ché definite diagmmaiv vf -valvulax regurgitvsivn

fin paviemaa suapgcaad vfi havimg fihis type defvmvigyo

Eat t B included

Serial No . NHI° 253

lo Clinic of Surgery3 o Bethesda

PHS a NIH


Calendar Year 1 958

Part A 0

Proj ect T itle : The Use of Inert Gene s in the De tec tion of

Left a to c aigbt circulatory Shunts

Principal Investigator : Andree G o Morrow. d o D o

O ther Investigators Eugene Brenne old,'

mo D o

RichardJo Senders , H . D o

Estelle R a Cohen

Fred Bullock

Men Peers ( calender yeer 1 958) Potient Deyo (calendarTotal year 1 958)Professional : 3

Pro j ect Deseription

Previous reports have outlined the usefulness of the inert

gas nitrous oxide in the loeeliz e tion of lett o to c rigbt cirucldtoryshunto end the advantage s of thin method over the de termination

of oxygen difteregg

e e be e been proved o In the pest year the

inert gas Krypton bee also been employed in this manner o The

use of e radioactive gee bee the odventege tbet enolysis of the

blood semplee cen be eceempliebed very t epidly by t ingle countingend the raoult s ot the otudy ere fineedietely eve ileble o

te sts were carried out in 1 50 petiente o The reonlto were the

gene as ttooe ot the mitred e oxide teot end the odventege s over

the oxygen method were senile t o

Radiation eetety coneidere tione constitute a disadvantage

of the redioeetive ge t but further refinement in technique willobviate this dreebeek o

3 at lwe Kr“ me ets will be

continued in pe tiente end turtter otndiee made at tee ters

incree eing its oete tyo

Port B includedm

Serial No .

NHI° 25&

l. Clinic of Surgery2 Sec tion on Cardiology3. Be thesda

lndiv idual Proj ect ReportCalendar dear 1 9 58

Proj ect Title : A Study oi Factors Influencing Digitalis Effecton the nefrsctoty Period of the A tt ic -Ventricular

lands

Principal investigator : totem: L. Frye , 2s. D .

Other l weotigatons Eugene Braunvald, Bl. D .

Elan Years (calendar year 1 958) Patient Days ( calendar“total : year 1 958)Protesslonal

Proj ect Deact iption

These patients with ent onic atrial fibrillation have been

studied to date , after discontinuation of all pt ev lous digitalis ,

After vsgal blockade by atropine , acetyl- steopnanthidin was

adminis tered at a cons tant rate . The acetyl- stsopbantnidin t e

quirsnent for slowing the ventt iculat sate to 60 was de teenlnedo

A fter control data had been obtained, the infl uence of the in

fusion of potassim , cslclm , and nor-epinephrine on the ace tyl

s trophantbidin requirement was determined , Eollowing the ln

fusion of. potassltsn, a slight increase in digitalis requirementwas noted. it use noted that the calcium gluconnte solution

alone signifi cantly decreased the ventricular rate . Further

administration of ace tyl- stropbantbidin did not re sult in any

noticeable synergism on toxicity. One patient has been studied

dnring a constant dntuslon of nor- epinephelne suffi cient to

produce a 20- 30 nn tng increase in blood neessut e , but no signi

ficant difference from control data could be de tected.

Pt ogosed cout oe 95 pro ject, A dditional patients are to

be studded usdng the constant infus ion of ace tylm stroplm ntndell-n o

En addition, it is planned to study the attests st fever and

thyroid hotness on digitalis requieenent . A patient with thera

peutie mynedema is being studied at the pt esent tine o Deten

ninations in a hynotnyt oid state have been cm pleted and sini

lae' studies in a eutnyt old and hypet tnyroid state are planned.

Serial No .NHI" 256

l . Clinic of Sutge t y2a Section on Caxdiology3a methesda

Kndlvldual Proj ect nepontCalendar “feat 1 9 58

.tle z An Analysis ot Adequacy of‘

fielntenance Dose s of

Blgttoe tn Eollowfing name M gltallaation wi th

investigates : Robe rt L. Frye , M. D .

s tlgatoss Eugene Bt aunwald, H . D.

(calenoat year 1 9 58) Patient Bays ( calendar

year

:sct tptlon

eat deal oi conteovet sy has centenad about the

const itutes an adequate maintenance dose follow

ltz atlon with a rapid acting preparation such as

lllctlon have been studied

lat tats to a level

one pattent sequined no acetyl- stt ophantnldln at

The other patient sequined several injections each

indicated that he had lost his initial status of

igltaliz atlon.

ct : Additional patients are to

regimens toe administering malo

lt is planned to studs;v pat ients

oi digttonfln immediately followingts of asshole and also patients who sece ive oes

taliz tng dose of digltonln 1 2 to 211; hours following

Serial No .NM ” 258

1 . Clinic of Sa ger};

2. Section on Cardiology3. Rethesda

individual Ft oj e ct ReportCalendat

'test 1 9 58

E'at't A o

Proj ect Tltle : Eemodymamlc Efifec ts of fiapzw Digitallz atlcm in

Cardiovascular t ow el Smtj ec ts

Principal lavestlgatot : Eugene Kelly , a. D o

Other investigators Eugene Btam wald, ll, D .

tit s , Estelle R , Gates

Mt . l’

t ed Bullets,

Mas Yeat s (calm s/st yest 1 9W )Total :

Ft otessioaal

Pz aj ect De sct lptlos.

Relative ly little t ellstle data has been svsllaele on

the effects of digitalis on cardiovascslat dynamics in subjectswithout: congestive beast failure , A ccaedlagly, cardiac output

and central blood volume were measst ed by the indicator- dilution

technic , toge thet with at tet lal sad central venous pt essut es

before and 4 5 mlae tes sites the lateaveaccs admlalsttatlon of

0 50 to mg, M ahala . Lett ventricular stroke work and

cat-alas output declined significantly is two of seven subjects ,

and showed so slgaltlcaat chases la the tem lmdet . ls. two

subjects , central blood volmse deceecsed slgaltlcaatly o

Peggosed causes as gt o ject : A dditional subjects are to

be studleglo From the data it appears as it those subj ects xc

ceiving only 05 0 rag. M ahala tassel s o slgaltlcaat hemsdynsmlc

changes , while those t ecelvlsg ago had met e sabstamtial

effects . it ls pleas ed to study seessel othet subjects usingmag M ahatma

Pat e 3 included

Serial No o NHI=2é 0

Emperfimencml flagral znmufficiency

prae sure find 1 1 animals . Systemi c

heigha iucreased, Ehis effect was magnified by pxoatigmine o

de

presaure pulse are planned.

Serial No .NHI=260

individual Proj ect Report

Calendar 1 958

Re esaJoCardie lo

m rm wa A . G and

Pare Mitre }. Insufficiency . Am, J.

99 0 «Bfi lyo 1 9580

Serial do .“HI- 262

1 . Clinic ef Surgery

3. Bethe sda

PBS mindividual Projec t Report

Calender Year 1 958

Pert .e .

Pre j ece m ele e Elective Gardziec Ai m ee wedded by deems of

Veneried lm weemen Gam e s

Principe ). Inve eedgeeee : Elem e. Weldheueemg BK. D.

M m So Ez fim waldg E3. 0 0

mum . P. Sem en . M. D.

M beril‘

. BlM dWellg N. Do

Andrew G. Mam et-79 Ed. a.

debere Whlde

Mm deem (eelemdex year: 1 958) M adeline new (calenderle e/ 11.2 yeer 1 958)

Peeieeeflexmel

Eleeel'

ee eeardlee 813m m is well eeeebllehed clinically.

M em e. fie we e fele met; saline preeeduee m eeeneed etamve eefigeu en

@f the efifeeee e zf em ewe ee ee re m e dy used ext-w eeding ee luu enea

weeeedwe efleretse end eeeeylehe lale e em eye seeddel eeeeeeeealmy.

em eeeebldehed memed efi e eeem z’

mg dime leeeee is by mem e of

venerficulerf fum e’

efien eew ee.

‘

E’

me peeeexeedee heme we e m ddfieed deem ewe eelgdnellydeoeedbed by sem efifi em el 3m ghee m eal vemeue t eem -

n m the

hee et; ezseepg eexeeeey ble e d flew we e dreamed mm e z eeeeve ir

and m en pum ped ehee egla e M eheedeee Flem fiee mm the pulm eery

artery. Pe eeeueee weee rew arded did we left; m edium an d eel-em.

fishe s. cerddee m em e. leffe wedge-lemm e filling pee eemrgeo endderedc m easure s were 2mm . By w ieg m m em e and tee svarying ee

e ddee m em e eat-gem m ete eeesld be eez releeed m m

m im e lefe veneefieelee filllvleg preem eee.

Elemeefiem cervee wee-e ebeafimed heed; befieee feed edger m eet .

‘E’fee eeeeee m e See med while we eefim l we e em eez ddwelem eryImm e rse. m elee eff em efi eezreee teem vexfled.

Serial no .“31 4 63

1 . Clinic of Surgery3. Be thesda

PBS e NIB

Individual Pro j e ct ReportCalender Year 1 958

Prb j ecr Tirle : An Experimenrel Enve erlgemlen ®f the Use o f

Redieecelve Ge e Seluelene in the Diagno sis of

Rigbee re e lefe Cerdlec Shaded

Principal mve erlgem r: Rebere‘

E. L. Lang9 M. D .

Geher leve eelgeeere John. dw we ldbeueeeo bk D .

willlem P. Carmella M. D .

weary Feleen

Leeeder drown

den deere ( celander yeer 1 958) Peelemr meye (calenderbete l yeer 1 9 58)Prefie eelonel : Rene

Other

Pro j ece De ecrlptfien

Eleven dogs wi th errdficelly cemerrucred rlgbro te o lefr

ebunrea :e e well as fire mermel dege. have been studied experlc

'

menre lly. Sex of theee animals were dtudied frem. elx ee ten

deys pe sm pereelvelys and five were studied e cueelya lmmedlerelyafter cemerrucelem ed the shunt . Perency e f the ebunt we e

proved by eecrlflcdng the animal ameedleeely efrer ere d end

eremdnlmg rbe ene eeeme ele . selime eelurdde e ef Krypren5 were

inj eeeed lure rhe cbewbere ef the righe side of tbe beer: ea

well ee ebe pelmeeery errery. Eeeedlerely edger inj ecelenerrerlel bleed eeeyle e were drawn er eveney deeded derervele fer

mdnufie e. Ebe ee eemple e were ceme ted e s wbe le bleed usiege cenmlneeme gee flew Gelgere fimeller rube . Km ewe enfimeleg fthe

redioeerlvley of ebe eapdred air was eeepled by means of e

Ge iger tube ineerred edge the edrwey. e. eerked difference in

ereerlel bleed ge e eee eeee wee preeenr berwe ee dege edeb ebune

cemerel dege . fibfie de b E 3 33 emrbed in rbe Quad reread bleed

edeple . flee mermmwm. eedne in eemrrele wee 286 come ts per minute

der able s mme le . En dege with ebwmee rbe eeeeee renged fume

1 490 fie 1 4 9 000 per mdmmee .

Em mermel dege vfi reuelly all @f ebe de j eeeed Kryptenss

de excreted by ebe le dge de lee fdree tremeze ebreegb rbe

pe lmemery clrceletlemo med vdrre elly meme ed id eneere ebe

Serial No .

“5 1 4 6“


30 Bethesda

PES -fiEB

Individual Pre j eet RepereCalender fiber 1 958

rare A 0

Ere j eet Eitle : Am Experimenedl Evalueeien of ehe Use of a

Beeempxe seien Chamber in the Eteeemene of

Cerebral A ir Embelfiee

Prfimefipel Keve eefigeeee :‘

Williem P o cereeilo mu D .

Gabe : Enveeeige eexe Ané ree G o fiereeeb Kb D o

Eeymemd weeeee

MQm yeee e ( celendee yeee 1 958) FeEfieee Days (calender

year 1 958)Prefe eeiemel: Reme

Gehee

Fre j eee Be eeeflpefiem

weder genez el ene eeheefie end le eel eneeebe eiea the eeroeid

ez eerie e ef dege weee inj eeeed eephefied week vexyimg emeunee e f

ear . After e series ef eeeexele weee ebeefined eeeehee geeup cf

dege we e injeeeedwwfieh aim and seem pleeed in e eempeeeeiem

chamber under a preeeuee efi 4 eeemepheee e . end elewly deeempreesedo

Eeeee bee been me dfififeeemce in gee meeeeliey in the central

degs end fie gee eeeeeed dege o flee eeeeee efi eix e deg can

eelereee vez iee ceneideeebly and ie fie ehe u

ghe ghee em enamel

mmee be weed ghee he s e were emifieme xeepenee fie e givee emeeme

ef exe c

flee use of a diffeeene meme

thereegEBéeé eefimei is befieg~

£mveeefigeeed end inj eeefiem of degs

ehz eege e diffieeeme eeeeey eeeh e s ehe veeeebxal fie beinginve eeigaeedo

Page B included

Serial Ne e“HI“ 268

1 , Clinic of Surgexy3. Bethe sda

RES NEE

individual Pro]ece RepomGelandet Yeas 1 9 58

Part A 0

R e ject H u e : An W ee-M em e ]. Enveeeige tien of We cerdiel

metebemm dat ing Geedfie c Aresse

Principal Enve etigem e : ee eeph We Gilbez'ea M. Do

o men Eeveeeigeeem Rebeee‘

E’. Lo m ega M. m.

Elem-

y Fe m

aueeell Welleed

Mew Yeee' e ( eeleede e yam : 1 953) m emes; Days (calenderm eek yeer 1 958)Pee ieeeieeelz 31 1 2 Rene e

i eee Be een pefien

Expeeflm m Reeve been peefereed in seven siege uemgpefie eemm eeeeeee e e eke e me eefimg egene end fin ehxee others

uefing eceeylchelime o

Gez diec eeeeefi; we e weneed m m the e meetsieg agent after

cewai e ccle eeee . we eeeee we e them epeeed end we cem etery

e seie m em euleeed. m e hearse wee then .peefueed fez m eme};

mim eee wieh firearm hepe zé m ez ed e meefiel bleed deem free donut

dege . Flee eeee e weee be eed en we fl ew eb‘eefie ed free eke

C G KQWM‘

y ewe s em ele befeee em e eeo m eem meeeene ef oxygena

sen-bee dfie zszldeo gluee eeo M eade eemg end m e etseedffied Emm y e cide

were eede em ehe pee é ueeee eelleeeed ew e m e eew eezy gimme

di m-mg ereeetse e s m m e s on {she fresh bleed in m e w ees-ve in

W gem mealfiee eflem emd seem dieefide peedzacedee g e e well e s

m u m mies; ef m e w heeeeeee glueeeeo leeeeee em neeefizerified

feeey secede mete ghee eelee lefsedo Relieving m eme)! manages of

eta-eats» ee ee eeeeeeeem we e eeem eed by peefueiem week. fl e sh

M u riel blee d cemeefiedng me az ee eeing egeeeo Beanies ea date

imdfieeee we eeygee en: w heeeeee eefileeeeien Gammam ee eeiue

{em-em;a be e dfie iefiehed eefim eeefiee de em eeeeylehelm e erreeeo

m em efien den-m g veneeeeelee fi brin eeeem fie egpeeem eelym at; of m e beefi ng mem e. eeefle eefifseefiee m em e be eccempm ehed

afieee eeeezmreee e peefie eeem eagle em m e eefimg egeee

miw gee o

Premeed eeeseoe 0 55 “

3geefieee: Flee s £ 631: fieeeee c ém eeeeneiem

ea? we eeeeee e eeefleeg weigh n ee dege m eeeeed by eeeeylchelm e

£ 8 l Qdo

m e E ie eleded

Serial No . NHI- 27O

Per: A , ( centinued)

Pro j ect Title : Eemcdynemic sendie e Buz zes flbae l Body Perfusion

Project Descripz ien

Limb flew bee else been recemded during hypcxie . The

circe leeimg bleed is deee tmz eeed in aha exygemeeet using a

mfiz euee end decree eed diec regatiemo Wish cecal flew

meinbeimed cenebeneo limb and eebel reefieaence decree ee

we ekedly efber 6 cc 1 0 eeee ee e O E bypeeie; bewevex9 the initial

xe epenee be e been veefieble .

Fumbbex general perfusion

end bypexfie seedie e

cc e eeeeee flew dexfieg pez fmeiem fin the eemefi ex- eplencbnic

exeee ; e eepeeeee seedy en hepemfic bleed flew dumfimg perfueion

is in pregmeee by ene eembee cf eke gxeup o Eb fie gleaned so

semeies ee abe pmlmenery ve eculer bedo

fibre edeqeeee cembxel @fi 602levels ie necessary9

pes tiee leely in eke bypeeie expexfimeeee o ewe Q2 cemeien

eleceredee eee under cee cexe ceiemb ea pz eeemcs in cenmecefiom‘

Wieb eeedaes em membxee e gee exebemge befing deme in mm. Eebeet

Bcemee'e lebeeegexy. fibeee eleeexede e ebeufid peeve ueeful in

be eh efiudfie e o é . j@im8 seedy efi eke pbyeielegfic eepecge of

membgene eeygemeeee use fie pleemedo

Eaxfi B ie cludedm m

Seriei Eb .

MRI- 271


30 Be the sda

ens e m

mdfivicbae l Ptnj ecz Regent

Calendar year: 1 958

i ecmmale Elecnlve Cardiac Argent muting new ]. Bodyfienfinefien fin ene Expez im neel Amine lo flee

neleeienenine ef Elevened Ennz ncnndle c Pressures

fie fige eerdlnl Penna-e end Pe ebnlegic m lmenery

chengen

elem Renee Bil, no

when Envennigeneee Jeeepb wo M lbeneb n Do

Edwn‘nd E , weeps BEL. D o

Bunnell Eellend

Rey antenna

Renew Whine

W illem Lnngblfin

Ellen Yenne (ce lenden yeen 1 9 58) Pnnfienn maye ( celendnr

yeer 1 958}Prefe eemnel Rene

meet

Pre j ecn fle ece-engine

in e eignlfifiennn neeeennege efi enlnele end neglenlsn

enbj eeeed en flannel cendiepnlnnneny byne ee end ele ceLve newline

e zne eeg pnlm nney cengeem en fie seen in we weenpem ewe pen-inch

n we e weenleeed been able ne e dme an eleveeed pniunnneny venene

pneeenre necnncleny en tine eeebnmne efi m neeeo m anning; m e

2522 1 (liege name annulled by blue c easinnonn

lemm e-

a mam end lefe neg-in].

final eff sending peefinelena enem a end

fine ext ent m:

cendlefieny n e e d ne peeeamne elevemlnnn in glee needle : chenbexo

Candlec feilnee and lung im age file not eccne in film s gm np o

Sinn er reenlee new ebeelnecl an glance nnfinmln in which n

ennefinnene neneeem ne e m fineanned ebm gbm e anneal end

necnvem o Ew eveng in dege w bj eeeed Be remedies enre en'

. M em e

cendfienem en m en eendietsenpy lane fin cbe engine ef Qu een,elgnalfilennt

’; nfieee fin lefin anneal end engine venenfieulnn nneennz ee

new nam e , En eddfifi en o linens: en s: enem ied eenplleeelene

Serial R0 .NHI° 271

PBS m E1 3

Endividnel Prt t Report

Celander fieer 1 958

A t ticle Perfiedfieml

M $8 9Jo g $3 0 9 Gilbexta ago W. » M m e E a E 0 9 A c G

Eleeefive cardiac Axxeee Tbtml Bedy Pexfueien: The

Releefiemehfip efi Elevafied EQQKQCQE difi G Pre seuxe e Emrimg Axre et

fie Smbeeqmens myeeez diel Fumeaiem and Eeehelegfic Eulmmnexy

Changes, gemz nel efi flhexeeic Surgemy, Eelc 369 Km, 4,

pp° 534 0 5429 cetgbex 1 958.

Serial No . NHI=272Part A . (coatinued)

Proj ect Title A Study of the Dis tensibillty of the Canine

ventricle Dut ies Diastole

?t oj ec t Besct lption

Thet e have been five technically satisfac tety sepet i

mee ts . The distee slbtllty ce eve of the ventt icle exhibits a

tlat portion, flm. whlch theme is a leege volume change asso

ciated. wt th a smell peessut e cheese . This is followed by an

intlectise at as esd- dtastelic peesseee in. the ne ighborhood

of 1 5 em. HZ O sedge steep pet ttse at the curve at higher end

diastolic pt essst es . This shape has beee . obsetved qe ite coe

ststee tly. Wes tt tcelst distessfibillty de teemieed dat ing the

infusios s t epineshe ine or e er - epte epht ine has not been signi

tlcae tly ddttet es t teem flee cos trel state . it is apparent that

the csteChol emflees also t esult is . gt eatet steaks volume and

steake were tot eey gives emd- diastoli c -

e olume . This neeld

indicate that the ieett epic acttoe of these drugs is attes ted

by more cemele te systsltc emptyfieg.

Em ediately attest death , dtsteesibtlzlty has not be en

observed to be sigsittcse tly different teem eut ing lite , sog

geeting that ves te icslst seles sttee ts csmplete during diastole .

fiesstt te tien oi the pet lcst dtem has been me ted to

diminish diasteltc dfistee sietltty. En two eepettmee ts , a pro

gt esstve teetesse fie diastolfic disteestbility sceet eed as the

expet tmee t pt ogeessed. Es one of these , este ephstee shifted

the pt esset ewt sleme cast e sass ts coe tt ol levels .

Pt efiosee t set se Additiseel eeseeteeets will

be caested out es . the effects st catechol amines . testis pess

set e , heet t sate , assets see symsateettc stmeelsttoe will also

tart 8 included

Individm l Pro ject M m w b

Pasadect Title Develomm t of am {mim icm m ly uc Method

Prim im l Imvestigatm : Roman Lo Barm an

F mJec-fe Description

gas «Masherge fieteetor

berial N0 0NHI‘ 274

Laboratory of Tecfinical :eve1 0pment

Bethe sda Maryland

Indi vidual Pro j ect ReportCalendar Year 1 958

Part A 0

Proj ect Title : The Biochemi cal Effects of Ultrasonic waves

Principal Investigator: Alfred‘

Weissler

Other Investigators

CooperatingUnitas None

iMan'

Years (calendar'

year 1 958) Patient Daye (calendar yearTotals « 8 1 958) None

Professionals 0 8Other None

Project Description

Prof; (Began J11 1 1 , 1 957)

The purpose of this research is to elucidate the detailed

processee by'

ahioh ulteaeonic eaves affect living tissues and

simpder biochemical systems such as enzyme solutionao In order

to study the role of free radicals in chemical reactions caused

by ultrasound in aqueoue systems” the sonochemical yield of hydrogen

peroxide wee measured in'

eeter'

ehich contained radical scavengers

in various concentrations and wee also saturated with either oxygen

or ergono The results indicate that hydroxyl radical is an intenmediate

produced from water'

by ultrasound; this radical and. perhydroxy1 radical

(formed in. oxygenated solutions) are considered prineiaelly'

re sponsible

for such sonoohemical effects as oxidation and pelymeri zationo

In a generelueay9 this re semhdea the situation in ioniz inga

radiation chemisteyg but there are notable differences in detailo

Quantitative comparison of Ultresound and gammae rags, with respect

to the amount of enemioal change eeuaed, per' unit of energy absorbeds

in three different reactions 9 showed that ultrasound is several

hundred time lees efficient e The three reactions investigated'

were

the oxidation of'

0 000d. fi’

ferroue aulfate in O oh M sulfuric acid

(widely used for doai metry in radiation ohemiatmy) ; the reduction

of M eerie sulfate in Ogle. gsulfuric acid; and the formation

Serial N0 0

Progress During Past Year ( continued)

apparatus is being rebuilt with automatic frequency control and seve ral

other electronic regulatory systems o It was interesing to note that the

flow curves showed a break in the slope which appears to arise at the

time the stream breaks into turbulent flOW o The region of the break is

compatible with. Reynold's number calculations for the turbulent points

but a more dependable confirmation will have to be made ° The development of the apparatus for' measurement of flow has so much in common with

the apparatus for broad line NMR analysis that the new apparatus is be ingdesigned t o allow both of these pro jects to utilize the same equipment °

Direction of Current Research

A s soon as it is currently shown that the NMR signal nill. measure

continuous flows steps will be taken to investigate the possibilityof using the induced nuclear precession as a marker to trace and. measure

flow; attempts will be made to determine whether or not the tissue water

can. be distinguished from the water flowing in the vascular bed; the

possibility of measuring the alkali metals in entire small animals and

the general prOblem of broad. line measurements evaluatedo

Part B included a NO

Serial wo o“HI- 276

differences o Light emission drops to a low value after a day or

:wo in the most dilute solutions (0 0 000001% luminol) which indicatesthat the luminol is slowly destroyedo

‘

Washing out the dissolved

air with nitrogen does not greatly reduce the light emissiono

Other chemiluminescent materials are known, such as lucigenin

(dimethylbiacridinium nitrate) , trieminosiloxene, and violanthrone

dye o None of these is both (a) setsre sdluble and (b) an indicator

of hydroxyl or hydroperexyl radicals ; therefore our studies thus

far have been concentrated on luminol o

Direction of Current Research

A ttempts are being made to lower the background count and to

increase the reproducibility s Solutions containing both hemoglobin

and hydroquinone are under studyg in order to find wh ether a favorablebalance point exists between the intensifier and quencher effects o

If satisfactory counting of carbone lh is achieved, the work will be

extended to tritium and to solutions containing easily oxidiz ed

substances suoh as sulfhydryl groups e

Part B included.

= No

Serial No :“HI “ 277

Progress M ing, Past Year (continued )

mm ther development will be attempted to obtain electrodes

preferentially sens itive to potassium and to calcium . Althoughthe letter will in all likelihood require e somewhat different

to develop true micro electrodes coated to within

of the top will be cont inued .

Serial N 0 0 5351 - 279

Progress During Past Yearfcontinued )

finite possibility of using this system to de

termine freez ing points of even smu er quant ities but. the

can be evaluated

on an actual problem .

Direction of w e He eeerch

To cont inue to devise end. teat new methods instruments

es the occasion eriee e to apply them to local reeeer ch programs

Part B include ci No

Serial No o

NHI‘ 279

Progress during Past Year (continued)

excited glow discharges n helium at an atmosphe ric pressure were studied,and experiments perfor med to determine what apparatus could best be used

to power the discharge and cause it to have sufficient stability to permit

its use as a detectoro

A stable discharge cox1 d be excited in helium at atmospheric

pressure by means of using a stabiliz ed source of radio frequency powe r oSufficient stability of this power source could be obtained by crystal

control of the frequency and using regulated direct current power supplied

to power the hF oscillator l A direct current potential is measurableacross the plasma of an RF excited discharge , Presence of a vapor other

than helium in the discharge tube causes a dim inution in this direct current o

When organic vapors of many ty e s are present in the helim'nthe de crease

in the dire ci current is directly proportional to the concentration of

the hydrocero ons in the helium The direct current obtainable and the

signals produced by the passage of hydrocarbons through the discharge

was sufficient to permit recording on a standard recorder without

additional amplificationo The direct current output was found to

increase with increase in temperature making the device more sensitive

at higher temperature , and permitting, its use in high temperature gas

chromatographyo Sensitivity to changes in temperature pressure and

gas flow rate was not sufficient to require extraordinary care in

stabilizing these factors e

Detenfiination of the ultimate sensitivity of this device has not

poem possible because of the difficulties in presenting the dascharge

with a known precisely small snow to concentration of hydrocarbon in

the helium Sensitivities have been obpainad repeatedly in the order

of 1 molecule of methyl myristato in 1 0? molecules of helium sufficient

to obtain congle 'te analysis of long chain fatty acids using a sample

f 1 0 fin . Sensitivity i s at leest 1 000 times greater than

them e 1 conductivity detectors 0

I

The de actor is simple in construction and has been adapted for

use with t conventional, commercia‘

.ly available chm atographs, with

increase intheir sensitivity, in the resolving power of their columns

caused by using smaller sample s, and is rugged and stable enough for

rel onths of continuous use wi thout difficulty o

Ancillary me thods for the mism analysis of fats in

for gas chromatographic analysis of constituent fattyinj ector to make possible the introduction of micro

d samples in precisely knuosn amounts into the

thout contaminating the oeluan and the analysis

carrier liquid, a sample introduction device

capillaries, the ir volume determined with a measuring microsco;e , and

a device constructed for introducing the capillary and crushing it at

the column inle t withou interrup tion of the flow of car rier gas ”

Se rial N0 0Nfifi “ 279


Serial N0 0”HI ” 281

Progre ss Bi t ing Past Year (continued)

examination) and this method of’

measuring surface area measure s the

surfz ce of the envelope of the clumps rather than ihe surface eng

Vbloping individual moleculoS a

International

é onforenoo on Freeging and Drying hold in London o It has also been

.writton up and submitted for periodical publicationo

thio pro ject o

2Serial No e

“HI- 282

Progress During Past Year (continued)

use of fro z en dried material for electron microscopy presents two

fwndamental problems : One is that embedding materials do not seem

to completely infiltrate froz en dried tissue 8 possibly because of

failure to completely wet the tissue which still has some re sidual

water of hydration no matter how long it is dris The other is

that fro z en dried material without subsidiary fixation does not

stand up under the ele ctron beam Both of these problems will

obviously limit the use of frozen dried material for critical ultra

structure studiesg, but on the other hand rapid free z ing offers

possibilities of correlating ultrastructure with rud imentary physiological

states which no other method doss o One problan in evaluating fixation

procedure s is the cutting of sufficiently small specimens and also

knowing whether these represent protoplasn in a more or less the

same physiological state o To try to solve this problen we have

started colonies of both amoeba Chaos Chaos and of paranociao

Further experimental work has been done evaluating the properties

of various coolantsw in particulars the Freonsg which have be en found

to be nearly as good coolants as isopentane or propane and are much

less hazardous to nos e The primary purpose of the se and earlier

experiments has been to analyse the conditions of heat transfer at

the surface of the couple being cooledo It has been found that to 1:

good approximation heat transfer per wait area at the surface is 55: on

by H TSa fig) » where l

‘is the surface temperature and

“

Ifis the

tenperature of the coo t o Theoretically H depends on a large mrber

of factors9 such as the velocity of the sample relative to the cabriolet ,a

its si z e » shape, as well as the intrinsic properties of the coolait o

Experimentally so to be nearly a constant for a Q: oncoolant » s o go H sec , dog, for propane and mils/ cm

2

$9 0 0 dogs for liqui d nitrogen q If H is knosn for a coolant it is

possible to develop an exact theory for the cooling of a soli¢ofregular shape without phase change and an approximate theory for

cooling with phase changs o By application of the theory to :aperimental

cooling curves it can be determined whether a phase change has occurre do

Such an analysis of cooling curves of 1 0 percent gelatin blocks has

indicated that for cubes less than 1 mm on edge g cooled ir liquid

propane there is only partial freez ing of the eatero

The primary problem in the next fen months is a corre late siz e ,number and occurrence of ice crystals as observed by electron microscopy

with the colorimetric studies described shoes 0

Part B included a

(A ttachment 1 )

Serial

Part Ba and Publications

Information Theory in Biolog ,Pergamon Press” Inc a, 1 958,

pp ° 3595 370 0

Serial N0 0“HI - 235

Progress During the Past Iear ( continued)

A n additional 1 8 percent increase results when mil Teflon is used,indicating that at this blood film th ickness }’ the membrane is still a

limiting factore Initial studie s with the double membrane unit usingl/ 2 mil Teflon show approximately 1 00 percent increase in oxygen trans

for over that obtained with a single layer of mil film e

A n apparatus for maintaining constant temperature and mixing is under

construction e This will accommodate electrodes for direct measurement of02 and 002 tensions e This type of measurement should pem it the use of

plasma on another blood substitute during testinge Refinanents in the

technique of using twin membranes are in progress in an attempt to over»

come the difficult problem of maintaining uniform blood channels o

Other materials}, such as silicone rubber films:a will be inve stigatedo

In additions a joint project with the surgical unit is planned in the

animal laboratory to study the physiologic aspects of membrane oxygenation o

Part B included a NO

a Serial No .

“31 4 8"

Program Baring Past Yearwm tmued )

equations and appropriate matrix methm for analyz ing such

3373t have been fievelopad . m g m em l theory has been

ahwm to give the m u wknown solutions for ayatw s which

consist of maven ]. uniformly mixed c m ; r: Utiliz ingthe them general problem of data amalgam and W 1 con

struction in 33mm with m m gte information have been

W M m 9 war}: has W m w ittemmp for publication .

Pracfi cally, the mm h m hm appliefi to the m m

of trace? data. can fatty m id. m tabolim 1m eollabom tfiom with

w e m mW im o Ew e , the mummy problem W to W mthe metabolic pam W of m atefi fiw fatty acid (UFA ) andchylomicm W igw am fatty (Twfi ) o The approach wasto m m m m fi lm of UFA back mm the Blam e. peel and carbon

M au de xxx-M eek by its m tgbolim m am each be related to flux

of UFA out of gm mam a pm ]. by m m tem l equation of the

if them m amas m in em mam , (which was r ?

testem way can be m ad ta em u-

be cw boma fiiwme M an on

M Laballefl UFA m the plwm m manage by mm m m e m email.

excretion of labellafi carbona-mm m that due to TGFA o mm em u

putatiom m m M iefi W augh M am m this scheme w ing the

aumra theory m m um -ms of m agnum of UFA am the Fraction of

TGFA whieh gam e s m m the plu m . peel m UFA before beingmamm al Obtained o M m qmem ly, in M ar m m m actical

cm mrwem of transm m fum fi cm in m aps of m am animals

or in clinical swa m the camputatwm m pr : for the IBM

by rm W ilm a

E memtly, QW M M of UF& recycling in fastingm exercising

subjects has; raised doubt about one of the M am mtal assumptions ,

W 1 31,mat all mm in. tha pm m l has the eam turnover time .

1 91 1 3 is w as“

hem checkefi em rm mmlly .

m magm a w to 51m m the gam ma}. W to

particulm problm in biological gy mm such the apglmao

m a mmm ea a mo

Report of Nhi Program'activities Mweee 31mg - Forgotten Books

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Transcript of Report of Nhi Program'activities Mweee 31mg - Forgotten Books