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Full text of "Report of program activities : National Cancer Institute"

-ANNUAL REPORT 
PROGRAM ACTIVITIES 



NATIONAL INSTITUTES OF aEALlH 



\\ A'l^lii 



CANCER INSTITUTE 



-KATIONALINSXmJTESOFeMLTH^^^ . 

PUBLIC BEAim.SERVlCE. 
■PARTMENT OF HEALiU^^ EDUCATION; AND ^'W^ 



^I>6r^ ^-^^cjr^^ «e:d^.t/.zC<^5. 



THE ANNUAL REPORT OF THE SCIENTIFIC DIRECTOR 

The National Cancer Institute 

Calendar Year 1959 

by 



Go Burroughs Mider, M„ Do 



J 

TABLE OF CONTENTS ^c/^ 

Page 

Introduction 1 

Causes of cancer 5 

Chemicals 6 

Mechanisms of carcinogenesis 9 

Viruses 12 

Polyoma or parotid tumor agent 13 

Leulcemia virus (Moloney) 16 

Mammary tumor agent 18 

Rous sarcoma virus 19 

Other carcinogens 19 

Host factors in carcinogenesis 21 

Cancers and their properties 22 

New tunnors 2 3 

Plasma cells and plasmacytomas 24 

Dissemination of cancer 28 

Polysaccharide research 31 

Protein and amino acid chemistry 35 

Nucleic acid research 38 

Endocrinology 42 

Radiation biology 45 



Natural history of leukemias 49 

Anemia and related disorders SZ 

Weight loss 53 

Tissue culture 54 

Cytochemistry and histochemistry 56 

Dermatology 57 

Surgical and virus treatment of cancers 59 

Virus treatment 61 

Pyridoxine deficiency 63 

Radiological treatment of cancers 64 

Chemotherapy of cancers 66 

Choriocarcinoma 67 

Endocrine therapy 69 

o,p'-DDD 69 

Screening activities 70 

Anti-leukemic compounds 72 

Alkylating agents 76 

Tetracyclines 78 

Miscellaneous studies 80 

Service functions 80 



NATIONAL CANCER INSTITUTE 
1959 

INTRODUCTION 



An increasisagly vigorous and productive research program 
prosecuted by the intramural staff o£ the National Cancer Institute 
during 1959 makes this siuziznary of selected accompliahxnents @ven 
lesa adequate tSian ita predecessors. New approaches to critically 
important problems in the study of neoplastic diseases have charac- 
terized the year's work^ Group enterprises have flourished to a 
greater extent than before, yet the talented individual v?ho prefers 
to work largely by hinnself deserves and gets equal encouragementc 
Ideas are born in individual xnindSo The communication of ideas is 
probably the single most important stimulus to productive scientific 
research; hence creation of an environment conducive to intellectual 
give and take beconties the most important function of iXianagemento 
There is no single or easy solution to this problem in a large organ» 
ization which depends for its vitality on individual initiative and 
intellectual curiosityo 

The virus oncology programs for exatnple, coztimands respect 
for its continuing high level of performance c A part of the effort is 
centered in the Virus Oncology Section of the Laboratory of Biology, 
but equally important are individuals in several different laboratories 
Mr ho are interested in developing significant segments of the total 
program by their own ingenuity and have only intermittent contacts 
with colleagues whose interests are 8imilar<, 

The entire area of chezxiotherapeutic research^ nauch larger 
at this point than the virus program, has been more strongly influ- 
enced by a few acknowledged leader Sp While this area is compart- 
mentalized to a degree^ there is a greater tendency for the individ- 
ual investigators to exchange information on a continuing basis. 

These examples could be extended, but simply indicate that 
research is people and that people differ from one another in ixiany 
important waye<, The organisational chart, so dear to the heart of 



the profeasiooal adsziimstrator, can indicate cfeanaels for conducting 
ordinary Rainess, but it caa stifle craative etideavor if it is taken 
as a blue prisxt iov the conduct af reeearcho Clearances, coKfirsna~ 
tions and authorisations required by rules and regulations oi the 
mammoth federal establishment create a set of circumstances that 
can lead to personal frustration, impede progress and cause the 
affected to seek ezTiploycnent elsewhere^ Substantial advances in 
decentralisation of decision making through the whole-hearted coop> 
eration of the Laboratory and Branch chiefs and other senior members 
of the NCI staff have helped in reducing a nusnber of these petty but 
highly sensitive problems. The roles of the Adxninistrative Officer 
and Administrative Assistant have been especially valizable in reducing 
causes of friction; yet inuch remains to be done. 

The size of the working group centered about a scientific leader 
can be determined by the leader* s ambitions, personality and research 
competence, hence the section becoznes the key part cf the intran-iural 
brganizatioQo The long range planning and business aspects of the 
operation require the attention of one who is actively engaged in a per~ 
sonal research prograzzi but is able' and 'ovilling to accept even broader 
responsibility -- the laboratory and branch chief. The last few years 
have seen increasing concentration behind those with ieadersMp capa- 
bilitieSo The head of a section has generally fared welL Some labor" 
atory or branch chiefs, on the other hand, have been ©o overburdened 
with so many different types of responsibility that their pes'son&l 
research, from which they derive a large degree of satisfaction, has 
been subjugated to the cosnmon good. It seems desirable, therefore, 
to reduce the aisse of some organizational segments of the Institute by 
creating one or nnore new laboratories or branches, probably during 
I960; rather than to persist in stultifying research activity through 
imposition of related but essentially non -contributory duties on the 
Institute's leaders. 

The death of Dr„ Jesse Philip Greenstein, first Chief, Laboratory 
of Biochemistry, on February 12, closed the career of one of our most 
distinguished, imaginative and productive scientists, a beloved friend 
and counselor, and a world leader in biochemistry and cancer research^ 
Dr^ Greenstein worked at a furious pace in 1939 when he joined the 
group that became the National Cancer Institute, and that pace never 
slackened. His interests were extremely broad. His mature judgirient, 
boundless energy, lucidity of expression and his firm though under- 
standing, even jovial, nature made a lasting impression on even the 
most casual acquaintance. He had a profound effect on the developnnent 
of the Institute's research program, which often extended into many 



arsaa of the institute's expaading activities. He was completely 
loyal, but very few people realise the intense and benevolent interest 
he s»howed in the personal problemns o£ bis asfiociateSo Dro Greenstein 
contributed most iznportaatly to aa'eas of amino acid, peptide and 
protein biochemistzy, including enzymology. He defined the concept 
of anaplasia in biochemical terms, and left behind a saonunnent, "Bio* 
chemistry of Cancer, " one of the most quoted works in the enormous 
literature of cancer. 

Dr. Herbert A. Sober has aucceeded to the position. Chief, 
Laboratory of Biochemistry. 

Dr, Oelbert Mauritz Bergenstal, Assistant Chief, Endocrinology 
Branch, died on September 12 after a long illnesso A close associate 
of Dro Charles Huggins, Dto Bergenstal joined the staff of the National 
Cancer Institute in 1955 after a brilliant career at the University of 
Chicago. He was a superb clinician with great ability in clinical inves" 
tigationa, His warm magnetic personality, his tremendous drive and 
infinite patience endeared him both to his colleagues and his patieats. 
Hie arguments were imaginative, his e^osition atimulating. Among 
his important contributions to the endocrinology of cancer are studies 
on the metabolic influences of ACTH, cortisone and growth hormone, 
characterisation of catabolic responses induced by adrenal and thyroid 
hormones, the effect of adrenalectomy on advanced prostatic and 
mammary cancers, and the effect of DDD on adrenal cancer. 

Br. Mortimer B. Lipsett has been appointed Assistant Chief, 
Endocrinology Branch. 

Dr. Nathaniel I. Berlin was named Chief, General Medicine 
Branch, a duty most capably discharged for five years by Dr. Cha?lea G„ 
Zubrod, in addition to his nnaay complex responsibilities as Clinical 
DirectoVo 

Two meetings of the Board of Scientific Counselors were held 
during 1959. Drs. Charles Huggins, Carl V. Moore and Eugene P. 
Pendergrass retired on June 30 froni the Board on completion of their 
appointments. Such events, though necessary, cause deep regret, 
especially when the relationships between the staff and the Board are 
as cordial as were those enjoyed with the original Scientific Counselors. 
Continuing excellent relationships are in prospect, since the new mesxibars, 
Drs. Hugh R. Butt, J„ Engelbert Dunphy and Jacob Furth, are just as 
deeply interested, understanding and cooperative as were their prede- 
cessors. Dr. Wendell M. Stanley relinquished his position as Chairman 



of the Board but consented to remain in active snembership through 
June 30, 1961, with Dr9„ E, K, Marshall and Philip P„ Cohen„ 
Dr, Cohen is the new Chairznano 

The Board of Scientific Counselors continues to sKprssa 
confidence in the research staff and its prograsn. Their discussions 
are most helpful. The differences of opinion that should character- 
ize a good and intimate working relationship are resolved by free and 
frank discussion to the ultimate satisfaction of the various parties. 
The meetings to date have required unduly inteni^e effort from the 
Counselors, who xneet throughout the day with various mesnbers of 
the staff and discuss program among themselves far into the night. 
Different types of meetings are being discussed with the Chairman 
as a means of reducing the work load. 

The increasing number of scientific meetings desnanding the 
participation or attention of the staff is gratifying in the sense that 
it confers some distinction solely for research accomplishment, but 
it is a^so disturbing because so much of the effort of our talented 
associates !»ust be devoted to preparation of papers. Program has 
sst-beeit-serlousiy damaged as yetj but the saturation point has prob- 
ably been reached. Foreign travel, though snot yet as comsrionplace 
as domestic travel, has becoine the order of the day. The problem 
is almost impossible to control as long as the upper echelons of 
government are internationally -irjinded, and in soine cases actually 
solicit the participation of staff Oiembers in planning foreign venture s^ 
Every request for support of travel to foreign sneetings subxmittsd to 
this office has had obvious merit in relation to research activities of 
the traveler, and the ability and desire of extramural organisations 
and groups to finance travel for mernbers of the staff of the National 
Cancer Institute have increased materially. 

Tangible recognitions of leadership in cancer research on both 
national and international bases have been accorded to snany persoas 
in intramural research^ Dr, W, C„ Hueper received the A, A, A.S, - 
Anne Frankel Rosenthal Award for 1959 in recognition of Ms contri- 
butions to knowledge of the causation of cancer in ir)an„ Ds, Harold L, 
Stewart retired as President of the American Association for Cancer 
Research^ Dr, Murray J, Shear is the Vice Presideiit of the A„ssociation; 
and Dr, Thelma 3, Dunn was elected to the Board of Directors, 
Dy^ Dunn is also President of the Washington Society of Pathologists, 
Dr, Soy Hertz was appointed to the Hesearch Advisory Council of the 
American Cancer Society for a five-year teriK, Dr, L, W, Law has 
been nazned as delegate to the World Health Organization's Advisory 



Comniittee on Cancer. D?» A, Jo Dalton is President of the New York 
Society of Electron Micros copiets. 

Am exhibit, "Tiamor Cells in Blood," by Drs. Jo F^ Potter, 
R„ Fo Kaiser, R, A, Maltngren, A„ W. Hiiberg and J„ C. Ppuitt 
received an Awa?d of Merit at the annual meeting of the Anierican 
Medical A|ssociationo 



CAUSES OF CANCSK 



Twenty yeare ox e^e^ience in carcinogenesis research was 
highlighted late in the year through actions taken by the governrijeat 
as required. by aiTsendments to the Federal Food, Drug and Cosmetic 
Act enacted in 1958o Several members of the institute's staff were 
most helpful in furnishing advice sought by various interested parties 
on scientific facts relating to general and specific aspects of carcino- 
genesiSo A meeting of some 25 investigators especially experienced 
in research on the causation of cancer was held during Decesrsber to 
deterxnine the extent to which agreement could be reached on different 
facets of the complex picture of carcinogens sis » A lively discussion 
elicited these ixnportant points; 

lo Carcinogenic property is not comparable to a physical or 
chemical property, 

2o Groups of compounds possessing cojnmoa biological proper- 
ties may be carcinogenic by icirtue of those properties, specifically; 
ionizing radiations, estrogens and goitrogens of the thiourea type, 

3c Determination of carcinogenic property requires judgment 
as to the validity of the experimental design aad its adequacy in 
relation to the known qualities of a compound and its relatives, to 
biological availability, to the reasoaabie interpretation of results 
and to the reproducibility of the responseSo Many isolated esperi- 
ments recorded in the older literature claiming carcinogenic proper- 
ties for certain chemicals remain unsubstantiated and are of question- 
able validity = 



4<, The potential hazard of a cheznical cozxipound can be 
estimatod only oa the basis of currently known facts and each situa- 
tion must be judged on its own merito 

Much nnore remains to be learned about carcinogenesis, and, 
indeed, substantial progress is being made in both practical and 
theoretical aspectSo We conceive our duty to be the accumulation 
of additional facts and especially basic principles which xnay be 
applied broadly to general and special problemso rathe? than to 
eznbrace an eictensive testing prograzxi of large nunnbers of com- 
pounds for purely pragmatical purposeSo At the same tiTne, how 
ever, we are glad within the limits of our knowledge to advise others 
interested in such testing which may well assume increasing importance < 



Chemicals 

The ability of some druses to induce cancers in mice only 
when injected shortly after birth has raised a question as to the rela- 
tive sensitivity of infant and adult mice to the action of chemical car-> 
cinogens^ Xnformiation advanced by Shubik suggests a heightened 
carcinogenic response to 9; 10 dimethyl-1,2 benzanthracene^ 
DrSo Margaret Kelly and iP„ W„ O'Gara have injected microgram 
quantities of S-methylcholanthrene or dibenz la, h( anthracene subcu- 
taneously into newborn S'ST'iss znice and xziice of strain CSH, Both 
respond by increased frequency in pulmonary tumors of the charac- 
teristic murine type recognizable as early as 16 weeks and becoming 
progressively more nunrserous thereafter.) The incidence of subcu- 
taneous sarconrias, however, does not keep pace with that ol the 
pulmonary neoplasms^ The same procedure elicits fewer tuznors of 
the lung in mice of strain CSTBU Other types of cancer have appeared 
sporadically,, This research is being extended to other chemical 
carcinogens, since the differences already observed suggest both 
qualitative and quantitative iznportance, 

Drs^ W„ Co Hueper and W„ W„ Payne have been cooperating 
with Messrs Ac C„ Stern and E, C„ Tabor (Taft Sanitasy Engineering 
Center) and with Dr„ Paul Kotin (University o£ Southern California) 
in a study of cancer production in mice by dusts collected from 
several American citieSo The benzt^aj pyrene content of each samsple 
has been analyzed so that the cancerous response, which can confidently 
be expected from the experience of Drs, Joseph Leiter, Mo Bo SMmkin 
and Mo J. Shear reported in 1942, may be related to a reference 
standard. Aliquots of each saxnple have also been divided into several 



fs?»€tiott» s'eps'eaeatiiig diff^g-igffit types ©£ a^oioaatic ©£■ aliphatic 
compsm&ds. Ps-oBsnin&sy ireeults indicate diU9f®uc®s in t2&« 
c&jrcinogeaic potency of th« fractions o? e^t^acts g-eHatizsg both to 
tha sous'fise o£ the stas'tit&g matsrial arad to the chemical OAtus'e of 
the f?%ctio2i tested, hi &t lemst two casea^ the %HphAtic f^actioais 
appe«s' to be more cfts^ciBOgemc th%& those composed ol A??omatic 
coisipoux^dSo 

Fustfees' experience with ceg-taia chromiam compoimds now 

peg-mita D?o. Huepes' to evaluate their relative carcisBCgemcity i&s 
s-atSu whett injected iatg'am^sculag'ly oi? iistg'apleuE'aEy. i® this 
descex&dio^ oddest calciwm ch5?®naat®o aiateg'ei caEcium ehs-oraate,; 
sintered chromium ts-ioxide,, ziii^c chromate» st^oati^ni chi^osnateo 
bas'ittxn chromate iwaakl aad lead ch!?omat® ((stois°ca5Pcitt©gemc;^o 
Dwo Payae iFepos-ts calcium chs-omatig axsd sixateg-ed calcium cfoiromate 
cas?cisaog«mc iow mice whes she@p fat is used as the vehicle €o^ 
subcutaneous iojection., 

Fg-actionation fey pa?°ticSe size aad chemical assaiysis of the 
iFesldue fs-om leached chs-osnite ©3P® ^©vsal that the sssialles? pa5?ticle© 
coastaiia Mghei? psopog-tioKS of hexsvalexat chromimnc thus ine^'eassjEg 
feei2> acute toxicity whea iEDteaiedo Removal of wates" soluble com- 
pounds fg-om the iresidue does not altes the degs-ee of cafcinogenic 
potencya according to Di?8o Payne and Huepeg-c Evidexatiy both Ife© 
discarded pasrt of the roast as well aa tJae desis'^d mij&tuge of chroiaium 
compounds inay be cas-einogemc haaayds in industryo 

Eairlieff 5?epos't« have secosfded the production,©! neoplasms by 
subcutaneous injection of a variety of wates' insoluble polymei?® 
studied by Df o HuepesTo He now has completed a study of watei? 
insoluble polyvinyl pysffolidoneso polyvinyl alcohol and dextsanso 
Sa^^omtas develop in those oirgans in which the injected £nat@iriai is 
deposited^ especially in the yeticulo-endoflselial systsmo Polymers 
of the same typeo but pa-oduced by different ps-ocesses^ possess 
different cai?einogemc potencies „ and neither chemical nor physical 
factotps of the molecules themselves can be s-ecognized to account for 
the diffe^enceSo These results seem ainaost identical with those 
obtained witts wates soluble compounds of the same genesral classesc 
The deposits ag-e easily i?® cognized in the tissues into which they 
wes-e oslginaHy intj^oduced oa? subsequently transpogtedo Possibly 
^e physical state within the cells may be pas-tiaJly responsible fos 
fee carcinogenic effecto 



Silicon® gmxn aed powdeffed siEie^- are maffi,, hnt @iiic©aa 
rabbea* produces ia@opl®sj5JS wfesm mtpctdtac^d iat® th® SMfeciat®ssie@ia@ 

Liiseai? ntiolecales ©f silleoise irufebes' &t-^ w«akly eareisasg^Eiie M 
bestc Pelyetfeylsa® isi ^^odcrately eas-ciaogsmc w&eai isaplsEited 
Si£bcista2seo^32.y a^ filra.® or discs but is laaetive as a powd^ir, a 
pxisicipl® e^sablislaed #®.iFii©ir by the OppessfeciEaeff® I©zf ©tfci^if plastic®. 
P^lyias'etihia^® us a aipossif @ also produces €©sm@ctive tisaiae tmooiaie'Sr. 

Hsated fats asid chiajpr^d fsods ay® kxaowE t© coat&ia ©«aib(8tafflic«§ 
l&M caxii ©licil a caKeeroMS fesposs^ m mieec; S&tnm yeai?@ ag@ 
Ds'o Hfeiirpsif becaen© mt®ii=est©d m Sfee poteiatial caiJciEogesjic hi®*ag-d 
fi?®m colfee C0s!ij^sxxipti©i&o Dg'o Mo Kisaifatsttia® wag aM© t@ idesiiilf 
teeisa fal pjwmnm iu c©fi@e sootSo so a lirnited study was iaaliat^d 
witsa ths €ii5©p©ir®tioffl, ©I vaffioms tn^snhms® ©I feg c©fl©g iadMStry-. 
I^r®. K.Mip.m-5«%affi@ asid Hweper cssaciud© that afomatic feydff©ca.rfe@5a§ 
■feifii p5F©si€if4 isa nao®l ffsasled e©lfeg bemsBS Ik ©sctjfemely low c©g£€©si» 
8.ifati©is.. OiSily c^g-taisi '"dark i?@asts'* wMcfe lorEsi ass ©3et5?esi«§ly small, 
ifactio© si ifee lotai pffoductiosi of coffee sjad ai?# eteifaeteB-igiicaUf 
iK&de by las-dly esjae©!?©® cemtaisi asay deteetabl® aj;ii©®s>l ©I kaewss 
e&iipciisoggssic hydroc&iybeaiiiit, Cofle® ceiagsjytarapti©®^^ t&eis, play® m& 

S&&IS ficous. €Qii«m roasting plaiats &w^ aa issapoifiast (somxsierteisl 
se.-tas'c^ el caffME!.® i® which beiaa 1^3 pytfeffife' is qwit© ®®lmMec 
Ds-Sc liwfffttsifflsse aisd Hwepes' mad# us© ©I this fact to @t^dy distrfbmlSeE 
6i Jte« e®s=cis0g©ia Ie the ^torsjacla ©£ the sa®ws©o Inters© fiaosfescesice 
d«»«fsl@p® iffi the Ftamesa of the stossiacfe^ wbea?© befiUB {ajf pyipeise is kaew'si 
I& elisli tarii.eg'i* wfesE fedi, b^t doe« r^ot appear lis th% glassdtalaj? rameosa 
Is '^Mcfe hmi&z, \&j pyfesse ®v©k€i§ is© caaeegous f©®p@isseo EairMigg' 
«s;«;p« ris?i®E£i« iFsp®rted by Dj?^ He L,. Stewart feav© sleariy showa tfeat 
^muM c-&wcm&gemc hyds'OC®t?b©E® eMeit som® adsBocarcmesma® wfee® 
iajfeet©.d diij@etty iteto tfe« giaadmlai!? gasitffic K»«.e©sau The ^mtire 
Sistpeg'iessee ffeeimphasize® t&« uieed ios study ©f gastJFlc sme^mti&un m 
yf/laSKOffi te c^Reeffigeisesis isi fee stosaacho 

JSSady ©i ttee causatioa ol g&itf ie eaELCss" has feees& IiaEsipi'S'ed f®?? 
^ack ©I ifeMable aisd reprodiaieible Kii«?tte©d8 fog" pjpodttcmg adsaosaffciaesr;.®* 
©I Ike .^loKSAcJuo Some progress has be^ss xnade eeceistly siEce Ds-g. *-' P- 
M&srm a«6d H. Lc Stewa-^'t feav« fo?sBd a f®w leaiosagi ia tfe@ glassidmlft-ic 
*j4,fK s.v-'-j &f ffais fed ^,j ? li?aoF©isytes!efeisacelaRudg. Wei?© 6fee saiae 



Issiosis seexa ii& humas> xnat«g'ial Ifeey would m&qu«sti@ssftMy h» 
diag3&ossd as adesocas'ciaozn&Ss but experieace ovmw ths yeag>s 
hA» ifeveaHed a siuznbes' of gacts'ie lesiGiss its mice that simulate 
gastirlc e%&e@s> but have ps-ovsd to be ffiOQ>Reop1lftstic„ It is es@«s%«> 
^alo theg'efog'ep to kave a clear pietus'© of tise bielogical bsfeavios" 
of gast3?i€ lesioi£S to iates^ps'st sigmfi€&sce of mos'j^ologicai shaiagcso 
Wbiie some of the Iseiox&e h£v# izsvaded Iks musculas'is m the Fate 
fed Zf, 7 fluos-^s&ylesebisacetaimdso no znet@st»ee8 have boea S'ecog^ 
mzedo AM of ^e subjects develop znetaetasizisg cm£>einoinAs @l 
the sniAll mtsstin^o %!%d some also have maligsiiant tuzraos'e of othe? 
sit@3 ifficludisE^ &<t salivas'y g&a2%dsc 

Du-o Wo Do Conway a&d Miss Eo Jc Lethe© have esEamiaed 
the puE-ity ol several corammseml ps'epas'atioi&s of the dyes yeJlow 
AB asid yellow OBo The Food and Drug Admimgt-ff&tioE limit® the 
quantity of ^--tsaphthyl^nusie in those piroduels to §00 pa'?ls peiP znEliots 
and osly oz^e of the tez% samples coi&tai&ed zno?e thaiK the pernUasibie 
quftRtityo 

Fous' mela&osnas have appeas-ed asnoag §1 gumea pigs painted 
wi& 9a 1 "dimethyl -io 2 bex&zanthyseeiie by Dg>o Jo Ho Edgcombo 

DffSo Mo Lo Hesselbssh fNlAiDI assd Ro Wo O^Gaifa sepos^t 
p^oductiois of subeutaneous fibrosaareomas in s&ts by mu^tljde lxye€^ 
tions of fast greea os> light greei&o. The earliest tumog>@ appeas- at 
sevei^ moisthso The sas'comas a?e readily tsraesplautable aad frequei&tEy 
metaatasizeo 

Mechanisms of cageisBOgeResis 

Study of ^e mechaaisxa by which N=2"lluoiFeaflacetaimde 
produces eai%ces>8 it& s>ats has sequis'ed identificatioc of excretion 
ps-oduetSs thei? sysCbesis a&d bioaesays cheznieal modificatioi^ of th@ 
pareist molecule mnd evaluation of careiitogeme potency of the aiew 
compouRidSo Much has beeu least&ed f^ona this apps>oacko &nd several 
aew c&rciuogens with iste^estiisg types of activity have been described^ 
such as 2o 7-0,u@s>emylei:&ebisae@tamideo The experie&ce to date ii^icatea 
that hydifoxylatioz^ of N-2=fiuos'eBylacetaimde lis poiiitioBS l^ Sj, 5 ©g- 7 
reduces its careinogemc potency;, whsch Dwo Moifs-is inte?ps>ets as a 
detoxification mechamssa os- a means of solubilization to facilitate 
excretiono Fluos'ination in the omega position of the acetyl gs'o^p 



pg'Cid.stse iKietttfiai IttsaoiFai v^eis led t® rafts, but fe«ps4-->ms9 wfct-jii 
l@d l@ does. 



hsisi al(»@ b@@;& studied by Dir. Mo^fis.. A high diesssf mt^« @l 

M=^'=lltt©S'eisyiacet&snaidec. M the- BianwraaiFy lesioas are ff©isj©v«td. 
sttS'gic&lly,. the ratis alsirnately develop hepatie aeeplasmSo C©Si=> 
"^«spseBye ifai& «atiag low dissary iffitake^ @l vitacai© B^ develop 
i«s£s&^i?s &i ih« M'i^sif bvtt ffiel @f the b^'eaalt., 

0«i.%iied .§twdi®@ by D?c H. M.. Oy«ff &f the snmtmh&Ummi, ©i 
l^ = «;-S-w«ijr'©ayliacfelairsi4de iro. Bmfial-© fats whiefe ate a sipyptepharo^tpiefe 
iimt wfer?- ffecerded Isa last y^^w''^ jpepcsjrt. Ttee ©fejeci4v« had feee® U&e 
jts'©«i^eiie'B ef tttKft@ff® ef Ifee Bjpissaipy Maddes" *@ fepoiFtcd tey J>isa!saiffi€o 
I'et aosife ©ccsirfedi.- !%« s&ua^g ps^^c-ed^aff^ does ps=®duee eafficer® ©? 

eherslcad diff«3P©iffiC«@ isi the iisges'elio© piredwcts of M-fa^fiimoreiByl' 
fecetasiaids 3g> &t irypio-pfeiss is tli© tw© stgaisiSo Addition e.1 Srypt<@pi8a® 
ii sw-t «&gesSial I® cajac®? prO)dy.cti@B<.^ Offic# aga.is»„ tfese g^ffietic essssli^ 

-Lis,, Uy^jj llffidfs difleg'eBceg in tte»« rfteiabeMsios ©I iffyps®^®® fey 

i^iffeatft!? at/satio® is aow b©S»af di^'ecled towasFds tiae l*te ol 
M«^»Eu<i!r'«aiyIl«.e';«ans4d«s Isa orgass assd tissues wfees"® the a^tnpew&d is 

H=2.»£l'4-C)!>?'S0fla(selaiBside labeled with G caa set b« esepilamed ©e ifee 
fe«i«iii ©i a3>4i£aeig €iTgat@d by in vite© nmsslpislalieavSc- Fuv&eTtm&s^,, 
c?-7t>thahydip«sjey/aled c©nspcstt©ds lo?fm stable coaaplexe® with a variety ©f 
iyi«€al i»&*,. 2sas«ff®dact4oss ©f a hydffoaeyl g sewp at a greater distassce iffi 
^e iE£i<sltsevil/ d«»e@ siot €osif@s> the sazne prope^ty^ 

■ H: Fc Blmn^j !P€vl«wlffif his visl «afepeffieBC« ia lfe@ :q*iAffit4t*tt*-e 
; . ■■ i"S!«'itS#S eaff'€to©gssie«i'e>, writes is fei@ ^ssost irecejat feocsks 



u„ 



owe jaaay affffl-^ft a* ee!?taijsi eojsuElttsios&s FegaE-dlag tMi§ kind &i e^wciuQ" 

"U i^ems elfsaff {hat wteate^eff t&e pw@cemm oi caaeerisalioa il 

&md cMStM&t fe« sep&ifa«ed tote distiEicI periods ^ 

"•Caa3i€©ffis&als®K i® ii® swne way c^gmtelatiti'e »- il dosage is at^^pedr, 
dev®l£fp«^(B8at e@iatiffiiw«s at a ipemipded jsaetg but tp#ed@ wp witia ,ff«ffi@wasl 

'Mtfeoagfe as^esffe is evici#ttc® ©I a gmaM degree ©I iFee©very„ 
tfe*& tt giight i® ©tf-eg-aW effect^, aiad easpelfflog^ffiesi© may fee iP©gas'd#d 

"&iE6C® tfe«r© I® ipecovery ■=- sued we iak# paiPtic«sl*5f aceoM®! oi 
fee- ««?i?li*ae« @f mis is the lallai?© of recipspocl^ s^t veipy Icsw d©©®-!?^!*® -- = 
it s©em# ^ftc®@ga5ry to e©85icllwd« tfeat fees'® is a lfeij"®*la@ld d©®af e at 

es.ffE&kil'-/ ¥'®i?y tew aiad is mot Mw^cUy me&snw^hlm fe©ca«a®« ©f lfe# MffliS#4 
Iii:*,jiy3ri,t ei' fee «)i®imal@o Tfaaso i«s m,Ky p5?actiea.Il ©»si@g, casfcissiaieffl^giis 
fe'j! •«sit»'a'#a«3rl©t ligM may b® eosisidgff'gd a® esseatiaMy ffi©©=.ifci5'«@h®ldo'' 

sliest* &ia©ffig eh^micaS caireiKiog^©® btet 11^4® tia© ©naaelilaliv© dat* 

WsSfe. Mjfo G. Ac. Soflesa CPg-iaeet.©© Usi^regsltyi Dw. SMm is 
»fes«if iiag she ^fimcu &£ siuglm doses of ^lltspavielet i?pjTad.iatios ©® m@«.i*« 
sfclK,. "Th^ ©ffigiisiili iffis^t i?8®d«g'si eell® isucapafele &i d4'ridiag» tewl £1^© 
4£j££«ff'(ft&& 'iQ®#s result iffi ©ormall /aaitsUe c@m2tsi mt .US daySc A serossf 
hfg.fes-j?l.&©|ic ire@p@ffi®e fellows km tfee epiderml® with Mffeeff pffolif@rg.&leEj\ 
cjfcie* fiteiaiis ^ff® lewad isa caiaeers R-es^tiffig frena repeated d©@©@ ®l ttls??®- 
-ifialefe,. The feyperplastic aP«sf3i©E®e lalls @£l with timeo Tfe« feigfees Ife® 
4i9.#fe« fehife gff®at«? J® tfe© siumfeeff ol ceUs destroy @d asd lfe« grmmtmw is 4fe© 
iaitial ifii.fft ©f c«Si ppollfes-atleffi! as measKsred fey mitolie iffidiee©. 



VIRUSES 



The p^oblazn of idsio^tifyisAg vis-mses that saay cause ca!3,ce?s in. 
man is <33ctg'exn'ely complesco One gieoup of iis^vestlgatoire und three 
individual scientists wos-king with diffageat techmquea have sca-eesaed 
naose than 100 clisaical eaacers for viguacSo Ossly oiae of thee© people 
has fouBd a clearly ideistifiabie viiFuso The recoveE-y of s. ntasabez' of 
viruses fis-om ts'asiepiaMsd expes'lzneaital t'oznos'S sueh as sarcoma. S7 
ie quite €ortunoi& and izucludes two xnew varieties of murine hepatitis3 
viyue fouBd Tecen&jf by DsTo Ro Ao M&imker in expeffimental leuhemiaso 
Eitheg" the e:&ncms9 that affect masK as?® b&sieaily different ia their ability 
to hairboj? passe&ge? virases or we ays working toward isolation with 
comparatively iatsensitive tools o Contiffittiiig ye -evaluation ol th© 
techssiques employed in the aees'eh fo? viruses in humaa eances-s pro- 
ceedSe together with etBejoijirageinent of individual imtiative in modifyicg 
techxdques of tissue esitsraction %nd tissue culture in the belief tha'J 
passenge? vis^ses should izahabit some ps'opos'tion of cancel's that 
affect mai&D 

DsPo So Eo Stewaift has obses'ved a pecttliar s'-sactioB m snosolaYe-r 
e»lt«3f e@ ©f fs'«shly isolated human ssTimos og emoz'yoffiic cells p2foduoed. 
by cojacentjrates of two differeat feum®.K cajEcess and the uJ'ines of th^ee 
childipen with neopiastie. diseases,, Focal as?eas of incg'esised gx'owrfch 
appeas" in the sffect®d cuitMre® leadmg to tk® i'o^'snatiojs of discrete 
denseg raised patcheso easily visible to the naked eyso Although 
ssiFial passage in tissue cisltu^e of the pg'olife^ative effect is steadily 
accomplished by use of supes'saatant fluidSo the cell fs=ee natu-re of such 
fluids ha® not been establishedo Filts-atioK in bact9i?ia-pj?oof Bjstoine 
OS? centa?ifugation at high speed d®sts?oys capacity of the fluid to trsr.iiniit 
the proliferative effects The sedixnent obtained fg'oni tlxsese ps-ocedu;?e,s 
s"etaim8 the ability to cause a focal pfoliferative g'esponee in new cviltnyr-^, 
and &u@ piropeffty is maintained aftes- E-epeated freezing at the t^m^es^ttj-^ 
of dry ice and intesimtt-ent thau'ingo The natns-e of the ag©nt t?es-pox~tsiblf' 



Natus-ally many tissue culture preparations o*" tisBv^e extracts 
have been injected into newboirn hamste-rs and micci, and the a.rr,.xn'.\lB 
hav® been obses'ved fof iong periodSo This experi-iance has sfe-emphnsii 
^e need fos" bettei? information on diseases occuyxring naturally Rmong 
labos'atos'y specieso Attention has been drav?n to a cuidotis but reasonsib 
common iesion of the placenta in hamsters which often results iv fetn.l 



i@9iosi8 of the iffilftstineii whieh ble«s>d profwsslyo 

Peiiyotai.^ vira« or p^agotid feijtmog ageat 

Some ^ozttfusioet ni*y b« caused by wse ®f th« aynoisyina poiyorraa 
viffus «®d p%3>otid tuzno? ag«ait les' aro age»t. wfeicfe pipoducea multipie 

pFi2R»ify xKeoplasms m mice az&d tumors i«ft ham«t«s'Sg iratii and rabbits o 

The cteemieai ©aftag'^s of polyoma vis^s has b««K studied by 
8ci«ffitists of tb« Sloafi=K€tterM&g lastiteste in collaboration witfe 
Dffu. Stijwaspt »ad Eddyo lEsf«etive imeleie acids psp^pas-ed after th« 
teehffiiques of Gie-?©^ aiad Sefeiramrri or of Kis-by mw« iffi»ctivat«d by 
dgoxyg-iboE^eiease but not by s-iboiiiiaclsase aa i»«as«.g'ed by effects 
osa cytopathic chaages la tissue cwltusre^ hersES^gglwtiaatioffi ©f gulssea pig 
erytfeipoeytcs aad tuznoF isadwettoift isa rsd^m asad feauasteffSo TIae iisfecti.vity 
©I Isitact viiTUsee is isot iisflueueed by eitheif ©f Ifee eaiaymeso Tfe® issfoetsvfc 
eompoa^fat ol polyomao tfeerelos?ea would appsar to be a deoxys'ibojswclelfe 

Earfy ia th© yeai? DUc Wo G„ Baiifield dgnooKStfated isitg'aisucl^as^t, 
iants^cytoplafisaic aad ©seiFaeeEulaff,, sheila tively iamfos'S3a<, ®pfe.<RFieai 
pasplicles i"? t© 3S rsfuElisvicifOES Ira diaisi^ieg' in lyrapbosna ceE@ gtrowa 
iffi tissue culture by Dwc Co Jo Dawa fos' propagatiosi of tfe© pas-otid 
tssjssor ageisto Uia,teociilat@d eultus'es eosjit^iaed jao ©incdJlas' bodies,, Tke 
dgscriptieiss piF©vid©d by Dfi-So Bsafield asd. Daw6 a^e coraparable to 
^© m.ea©«!Pene®Rt8 x'«po2't®d last yeas by Dso Ho KaMes' aisd coli#agwe» 
©E isolated pas-tlcles potsesismg polyoma activity whess allowance i@ 
made foff tfe« diif@r@iiBt laa^tfeods of prepaffati©»ia DrSc Banfield and Da.we 
CGiasidsiP feat p5?©pagalio© of tkm vii?um is predonaiisaxatly iffltraaweleaifc 
Tte*-' aM.g'&©«cl®ai? loeadizatio© of polyoraa d®r»offiSts?at©d by UuoE'escgaii 
fe,54ii^% techalqugg i». tissue culfctts'eig ©f a Sympfe©3ma» mm fspospt^d by 
Xijpg. Ho A. Malmgrsffio A. S,> Rabnoa aisd Gtasacag-lo RabotH ((Visitisig 
Sci(e®t4@l))a supports that viewo Tfee^e workerst, howev®?,, feave eofe 
fe©«^ a.feS@ I© dessnoiasts'ate poiyoaaa virus isa pacotid glassd tumos-s by 
llttsregegisi antibody teefeaiqueso Otfees>s have gfeowK. that isot aM teaaora 
^■S'Q€m€Md by poEyoma coBtaiis detectable aEaottsat® of tfe© ^ims wfees tissue 



Dr„ Bobert i-ove has applied Ms sensitive toluidiae blue 
molybdate isiethods for the dejmoBStratioa of intracellular ribonu- 
cleopyoteins to tlie study of tymphoma cslla infected in vitro with 
polyoma. Neither nuclear parachromatin nor the nucleolinus can 
be demonatratedo The nucleoLus is enlargsd, and vacuoles coiitaiiairig 
a diffuse aad condensed, abnorinaUy staining forsn of ribonucleic acid 
appear in the nucleus. The nucleus first enlarges, then shrinks and 
eventually disintegrates by kai:yorrhe«;i8o Dr„ Love has not been able 
to identify a deoaryribonucleic acid in association v/ith the newly 
fornned intranuclear ribonucleoprotein by histocheinical procedureSo 

Dr, Babson and Dr, Ruth Kirscbtein (DBS) have produced 
intracranial sarcomas through the intracerebral inoculation of poly- 
oma virus in hamsters,. These neoplasms appear to arise from the 
pia mater and the adventitia of the meningeal and cerebral blood vesse! 

DrSo Stanton^, Stewart and Eddy report that injection of polyoma 
virus into Swiss mouse fetuses in utero late in fetal life results in 
earlier death and a higher incidence of tumors than itijection on the 

first day of extrauterine life. Mica infected as fetuses develop no 
new histological types of neoplasmSo Some of the mice, howe^-arf 
born to mothers reared in close pi'ojdmity to adult jnice known to 
harbor parotid tumor agent had a significantly lower incidence of sali- 
vary gland tuimops and "renal tubular lesionSo" A comprehensive 
analysis of histological changes by Dr, Stanton reveals that soib© 
cells in the infected rrice hypertrophy, forsn intranuclear inclusions 
and degenerate. The neoplasnns ia the series are composed of hyper- 
plastic epithelial elements and proliferatiag or differentiated ceils, 
liznited in their capacity to invade surrounding tissues or to mslasta- 
sizoo Some tumors regress; others kill their hosts by local growth 
in vital organs o 

In another study of polyoma virus infection, Dr„ Sarah Stewarts 
Dr„ Kahler and Dr„ Stanton have studied the tumor® occurring with 
limiting dilutions of virus culture preparations subjected to differeri- 
tial high speed centrifuga-feioa. All fractions produce tm^nors, but the 
greatest activity is found in sedimgats produced with forces designsd 
to contain the greatest quantity of the agento Agsitis the variety of 
neoplasms produced by the fractions is similar o Ar^lysis of the 
tissue reactions that follow nr^inimal doses after long-term study 
reveals znany non-specific lesions in older mice, especially in the 
kidneys and salivary glands^ resembling either abortive attempts at 
cell proliferation or iregression of mimsmal lesions » Older mice do 



develop a significant numba? of neoplassxis, particuiajs-ly feo»8 tusnora 
that show clea? enridence of the capacity 60 irivade and snetastasiaeo 
Dr, Stanton cotnmeats: 

"The polyoma virua-tuirio^s ser^a as excaptioaal e:sperisnental 
toolSo The wide range of species that respond and convenience in 
working with ssnall rodents constitute an advantage ov^er chemically 
induced tumors both in the short latent period and opportunity of 
following the progress and beha^or of the tuinora in relation to virus 
as detected by new virologicai and morphological techniques. » . o o o c <, 
There is good evidence to indicate that induction and progression pro- 
ceed in a slow atep-wise fashion which znay allow separate study of 
the two most important attributes of t^unorSe namelyp cell prolifera- 
tion and the acquisition of aggressive behavior patterns by the proli- 
ferating cellSo" 

Perhaps the most significant part of this statement is the recognition 
of phases in the evolution of the cancerous state among neoplassns 
not associated with known endocrine functions „ 

Especially important to the search for viruses that snay cause 
cancer in znan is the ability to recognise a cancerous change in some 
foreign species or semi -artificial systexB such as tissue culture. 
LASt Year's report referred to the most interesting changes produced 
by parotid tumor agent in salivary gland anlagen grown in vitro by 
Dto DawSo He now reports that increasing chronological age to as 
much as 15 snonths does not diminish the proliferative response o£ 
submaxillary glands of mice to poiyozna virus in tissue culture. If 
anything^, the reaction, which mizsiics neoplasia in its morphological 
characteristics, is more pronounced. The altered cells still fail 
to produce local neoplaszns when transplanted to genetically appro- 
priate hostSp although evidence for virus transfer is clear. On the 
other hand, infection in vitro of salivary gland rudiments in the 
earliest stages of xnorphogenesis produces little in the way of a neo- 
plastic response, 

Dr, Dawe also reports that growth of adult parotid gland in 
sponge matrix culture results in digestion of the sponge by a protease 
secreted by the cells. 

While Dto Ko K, Sanford has been able to propagate polyoma 
virus in a clone from normal C3H miouse parotid tissue and in a long- 
term strain of C3H embryonic fibroblasts, two strains of C3H adult 



16„ 



fibroblasts and one strain of Chiness hannster cells, although 
supporting limited maintenance^ show no conspicuous effects of 
infect ion „ 

The failure of certain newborn mice to develop tumors when 
injected with polyon^a virus mays in part, be due to the presence of 
antipolyoma antibodies in mothers' nnilk transmitted to the nurslings,, 
Antibody titers as measxired by inhibition of hemagglutination are as 
high in milk as in sera, according to DrSo Stewart and Eddy„ Similar 
reasoning would account for the failure of naturally Infected stocks 
of mice to develop salivary gland tumors so characteristic of poly- 
oma virus action. Dr„L,„ Wo Law reports that mothers whose blood 
contained hemagglutination inhibiting antibodies in titers greater than 
200 delivered young which resisted the oncogenic action of the parotid 
tumor agent, Foster nursing experiments conducted by Dr„ Law 
point to the transfer of antibodies in the nnilk of infected miothers and 
also suggest that antibodies may be transmitted transplacentallyc 



Leulcennia virus (Moloney) 

The murine leukemia virus isolated by Dro Jo Bo Moloney 
from transplantable sarcoma 37 is beii;ig studied intensivelyo 
Standardised techniques based on differ-ential centrifugation at alter-« 
nating high and low speeds or on bacteriological filtrations combined 
with differential uitracentrifugation yield equally potent preparations 
froir. sarcoma 37 or from transplantable leukemias produced by the 
viruSo Leukemic spleens or lymph nodes are excellent sources of 
the agent, but the liver is a relatively poor onco 

BALB/c strain mice have been used by Dro Moloney as a 
reference standard in studies of viral leukemo gene sis. Susceptibility 
is retained for at least five nnonths, but the latent period is some- 
what longer among older miceo Mice of strains CSH, CSKf, A/LNj 
I, RIIIj DBA/2, C57BL and randomly bred Swiss mice all develop 
leukemia fronn virus infections „ The incidence approximates 100 
percent and the latent intervals of leukemia production do not vary 
significantly excepting for strain C57/BL which develops the disease 
in only about 30 percent of infected subjectSo The F, hybrids obtained 
by mating each of the strains noted above with BALB/c mice respond 
to leukemogenesis as proinptly as do mice of the reference strains. 



17. 



Potcmcy of the virus has been increased by selective virus 
pass&ge in vivo o Alter eight passages the latent interval has 
decreased from 6,4 months to 10-12 weeks in all mice tested, 

Dr^, To B„ Dunn reports that this leukesnia is morphologically 
indistinguishable from spontaneously occurring lymphocytic leukem- 
ias in several mouse strains „ The lymph nodes^ spleen, liver and 
thymus are characteristically enlarged and ir/iiltrated with typically 
lymphoid cells that also involve the salivary glands, kidneys, lungs 
and meninges in the advanced stage of the diiSease^ The same histo- 
logic pattes'n is seen in all mice estamined ^'/hether the leukennia is 
feranssnitted by cell free preparatiosas or by leukemic cellSo Blood 
from the leukeznic znice presents moderat'£! leucocytosis and anemia o 
No neoplasnss other than those of the lyniphocytic type have as yet 
been observed in any of tha test animials. Leukemia can be produced 
by Dfo Mol oney' 8 virus by subcutaneous, intravenous, intracerebral 
or intraperitoneal inoculation. 

The leukeznia virus remains stable at temperatures below 
-50° G for long periods of time when prepared in citrate buffer and 
withstands lyopMlisation without apparent loss of activityo Biological 
activity is retained when the virus is e^osed to 37*^0 for 90 minutes 
but is lost at eiqsiosure to 56 C within 30 sninuteSo 

* Dso A, J, Dalton has examined leukemia® produced by 
DSo Moloney's virus under the electron microscope. Particles 
representing t\& agent belong to morphological class Type A 
(Bernh&rd)» have an eternal diameter averaging iOO millimicrons 
and contain an electron dense nucleoid of 48 millimicronSo They 
are not particularly abundant, but may be found in spleens, lymph 
nodes and thymuses of infected mice and are found most frequently 
in the intercellular spaces and only occasionally within intracyto- 
piasmic vacuoleSo 

The Moloney virus is difficult to propagate in tissue culture, 
Dr„ R, A, Klanaker presents evidence that the agent can be maintained 
in vitro , but its behavior is not entirely clear at this time. Ameboid 
cells found in cultures of liver, thymus, spleen or lymph nodes of 
leukemic mice seem to maintain the virus best, while cells clearly 
recognizable as lymphomatous support it little, if at all. Although 
many other feypes of cells have been used to propagate leukemia 
virus, activity has usually died out within four or five serial passages 
in vitro o 



in-iJDwna sera prepared ircxn rabbits jf'fii'siv^ing nmltipt^ inje';:$i\>?4® •:<( 
the leukemia ¥'iru« do cQni!,ain oeutr.&ll:^'ing a>,nsibodieSc A dlulg«n«: 
aeairch by Dr= Mo A, Fink for other seTo log lea i -reacts c.«i« w&ti'te 
migkl b© tised in epidemioi^..glcai studtfe^ Smie b<f®n «BS«eci8«i«Jtuio 
Tlie agent does not fiat cojmpieir.ents noi- does it; cause fejtsnaggiatiB- 
a.tiouc As a matter of £act» jmost of tfe« oncog«F»ic virua^s are; iraihe r 
weakly antigenic^ Polyoma is an exceptitmc 



Mamnrt&ry tumor agent 

Dr^ Ho Bo Aaderifont rec^Btiy described the di*ap:p«.A4'S«i..« o^i 
the mamznary tumor ag^eni irom two fem&i® m4c© ol stfAitJ Billi, 
Eacperitnenfee designed to analyze the respective roles of tte -^Iru^ 
and the host in thie pfe@noBo<g«.o7i have included production. «f ^otsb, 
C3H and Bill swice kssown respectively to carry mnd to be i^tM of 
the agsnto These stocks ha%?e been inteybredo Ninety »fiv« perceM 
of th« agent -free C3H snice e3«pos«d to th«ii" own maiRDmary turrisr 
inciter develop tumors of the br©a®to Brg^eding decreasess ste 
latent interv'al but has no effect on the Incidanceo BUI milk m CIHC 
mice pfoduces a smaller number of »s,mmajry carcinoxrvas but Is 
transmitted serially through the strg,ifi.o It is already mp^ariirtit, 
however,, that soine RUI Rsice resist Ifee caar cinogenic effect® of the 
C3H agent. While the RUI agent is much lesi potent than Ifee C3K 
mammary tumor inciter» the constitution of the host i« aliso lir^por- 
tant in determining carcinogenesiSo Dr^ Aiedervont's earlier afe«€r- 
tfatione on the enhanced activity of the mammary tumor «.g«rit 
obtained fponi wild mice and propagated through 20 generAilons in. 
BAi-B/Cs coup'ied with Ms other studies on the dieapp'ss.ratics oi C3H 
agent from mice of strains C57BL and I, take on added significance o 

Further work along these lines ha.8 been conducted by Bfr, W, )£.- 
Heston in hie studies on genie influences associated v^lth th«! propn 
gfttion of the mammary tumor incite ro Segregation data obtained 
from the mating® of strains C3H and C5?BL indicate tha,t the %gent 
can be propagated in the presence of one or several genes rather 
thftn being under the cosstrol of a single geneo A fematle nr>ouse lack- 
ing ».ny of the genes required to propagate the agent does not e-Iimin'^ 
ate its but the virus does not multiply and is lost in subsequent gener - 
ationSo The quality and quantity of mammary tumor agent vary then 
according to the number of genes reqwired for its propagation and 
their availability to the indi'sddual coaceraedo 



Dt S^.f:Xor<i -^nis Urs, ijuna a»5c! A«?iervonl feport® the 
peraisteioice of ai&xriw^&.ry turr\~)r trxcitcr In cuituffss* oj c©?i&in Cis:!* 
for more than as. year, Actwtty is l:>Bt if %hm csiis lake un a sArcow; 
atous sppearanceo The usual atTuctujral ch«»i=a,ctierls$lcs of yr.oi*^© 
.marrjKsaffy e&rciao.nnas are maintained when the ceils grow in sersjrs 
8uppierne»l:©d. chemicaliy defined media but are tosit wfes^n tfe# im«d ' 
ium contains serum and chick embryo esctraeto 



Boua ga ycoprta ^i^ug 

Es:»,)mi nation of physicals chemical and ijr.muno logic prop© file's 
of the Rous sarcorxia -^drus .require® a highly potent sowrc© of rsrialef - 
ial„ DJo W„ Ho Bryan repog-ts a four-fold lacrea«g in activity during 
the past year in viyal preparations obfeained by sei®etw« ®€iris.i passag!?. 
This represent© an incirea®© of 100 time* the potency of th^. feeeit 
nriateyial available two years ago and rr^akef purificatlos Madieg 
practicaio The intranuclear and intracytoplasmic t.>c«.tion -.ii Hous 
virus antigen in Rous earconms h»& been deir.onstrat^id by Ors., Ma.lmgfef. 
and Fink using f iuorescent antibody techtiiquas., 

Through th© co..-pe*'ation o£ Dr.. Jo Ac Eeyniers fG^.rn-^ free 
Life Research Center^ ratsjipaa Flor jda)p Dr.. Bryan Isms at gas red 
a number of Japanese quail (Coturni» Japofjicj&i for use in E.:«i.v# 
virus reg^a-fcho These birds pro'^e to b# ai?s 8ugc«plib\@ to tuKior 
productioa as the itiost sensitive chickens yet iav-estigatedo Yields 
of virus are fully as high froir* qxiail tumors aa £ron< chicken g.amofs. 
The @m»il space required to house the Japanese quail, which a?^« no 
larger than fi^re-day^old chickS:, is a considerable adyamage. 



Other carcinogeni 

While much i« known about the qxiaiity of caresnogsnic responses 
to ionizing radiations, much remains to be learned concerning the 
mechanism of their action which can be approached through n-.anipu- 
lations that increase or reduce the frequency of the regponse. 
Dfo Law, particularly interested in X»ray leukecnogenesiSe described 
a .reduction in the incidence of X-ray induced lymphocytic ieukemias 
in thyme ctorr.ised CS7BL mice some years ago., The normal responee 
couid be restored by subcutaneous transplantation of thymic fragments 
in which the leukemias apparently origitiatedc Last year's report 
referred to the failure of thymic tissue placed intraperitoneally within 



millipore diflusioiti chamber® to restore tfee capacity ol thfx- 
rrici; to develop i€^-#mlae^ mitmr X=ir?&di.*lioii« T'h« tfc.jysnic tissue 
wilMa fehe cbatnbes' does ttot becutr.® ieaksinie in either CSTBj... niice 
or in hybrids produced by mailing C§TBL to strain A„ N«^#rtfe®les^„ 
to 30 percent ol th^se subjects ds^««lop s'etlcyl^F neop:'asrs;S ol Dy^ Due-: 
type A aad B,, No plasma csll neoplasmii ha-v© been fousi;d u««l«gg' these 
conditions o 

Dto Law now report© the pre%«stios3i of nsoplM^tic cimti^rsB tti 
thyroectonmized xr.iee bearing subcuEaneoia* thymic grafts wh^& 
isologjug teone rasarro'^ is giw^n i«inm«diste!y JfolJawing she Isst of 
four weekly doses of total body X= irradiation., Tb@ iocldeKr.g •■>{ 
reticular neoplaSCTsa other than iynAphocytse ieukerrit* i* not a^Tfecterfo 
Chimeras produced by injection c*£ AKR marrow ittto Irradlmted CUM 
nniceB of Uk^m or Bittner Sttblineg .lee© than 1^ hour® alter Msth dc:=5'/« i ..-p 
th® san:i« incidence of leukemia® ae do tb® controlSg nor l# tfee lewksiri.v*. 
incidence influenced by trAn&pia?5tAtio«i oi AKR fe&yn-sic eI««u«„ A 
wjost interesting obserHration^ feowe^ert is a frequertcy of url?&t@rai 
parotid feunxors, approMmating lU p«rc#nt a.?r;ong C.3.H/Bi nnc^ r©c:.»i- 
tis»ue IroRTi AKH donor© known to be iittected with pojyotra ^Irus at 
dsxnonstr&ted by HI ajstibodi^s^ 

Mis© Do Eo Upboff and Dro t.-aw lm^@ <?««nded tis«:ir attempt® 
at ialterlng the high incid©nc« jf leukeroi* in itrskdi^iml AKK snicig ta 
include the use of m&rrow frow; four straans tba.t ar^ catr-patibie at 
th® H-2* locuSo The restiit* are loconjpjetes but jtr.'sat of the i*?u.ke.ir. - 
i&s arising to date in mice iifradi^,t«d s».n«i protectsfJ wbi^K yo^ng fea.'^-'is 
p!eo"»?ed to originate in donor cells e^isn ebtmgfe they v^';on.ife frorso strainii 
with low frequency of spontaneous ieuI^emtAc Marr-vw tr^iiitaient m 
AKR mice irradiated at aiat months uf agg bsa little effect on tfe??. 
leakenrjia incidence^ 

Dr„ Bo ^n Swarni repirts hemai«gioeiridwth.««i-;.)Cfli4iLg oC tha ii"s'^r 
and spleen in two of three ra^l>bit® recei^ng two lisject^an.f «ach of 
3 n-'io of thoratrast (coUoid&l thorium dioseldislo the <«<!iioTsi! sppearfed 
At 36-3? months and the other nonc«KC«srous aniniia i, 4i€id. al iv months, 
A gingie intravenous dose of 3 rr.!., produced no neoplasn-.s in any ol 
9 rabbitBc, 4 of which sur^ved at 1©-%"^ *'■ — .-..t^f>»'» 



^I„ 



As better knowledge of eonatituUonal factors concefa^d with 
fch« carcinogenic process iB acquifed, thi^ iatejf-r@iaiion«hips are 
more properly discunsed in association witb specific responses 
which they modify under controlled conditions. The priacipai iden- 
tiliabie gemttic deterrr)ina.nt of the cancerous response seen:.® to 
reside in apecifie organs or tissues from Ihe work alifeady reported 
by Heaton, Gardner, Kirschbaurn and others. Two strains of mice 
may be almost completely resistant on the one h&nd, and ©xquiftitely 
ftuacepiible on the othcsr to production of a, given anatorrdcal variety 
of cancer under readily controllable condilionSo In what w*y do these 
organs differ? The morpholog4«t, well equipped to study lM« problem^ 
arrives at no definite conclusions^ The biochemist is confronted 
with large number© of celts of which an almost insignificant, fraction 
its- destined to become cancerous. The problem doeg not g©eit> in«up- 
erabiSp however^ when one considers the a'spailability of ailcrochem- 
ACS) techniques a» developed by i.inderstrom-i-ang, Lowry and oth-^r®:, 

OSo Heaton ha@ obser%"ed an increased frequency of hepatort as 
in C3H mice diuping recent yearSo He ascribes this in part to gesselic 
cha&g© in the strains but also to change In the diet in fels animal 
colony,, since mice eating a standard eomifnercial laboratory ration 
de^eJop sig?d£ica?5tly fewer tumors of the liifer than those eating 
a for.i">uia a»p«ciaily compounded by Dro Morris for «e# at NClo The 
presence or absence of the mammafy tumor agent In the stock has 
rxo is}Uuenc« on the incidence of hepatomas,, 

The influence of the lethal yellow {Y ) ge?je on mammary tumor 
d®'??iiiapmignt studied by Dr^ Heston is. manifested by a prolciaged 
latent period in idrgin mice of agouti color as compared with those 
ha^Rg y®l ow coate^ Both color types are obtained by mating CJM 
aad Y strain© since the Y gene can be propagated only in htetero- 
^ygotfeSo Th« difference in latent period would seem to be r©iat®d 
to «or/-t hormonal influence because it does not occur in breeding 
l«:?rai@Sc The Y gene also affects hepatoma incidence which is 
grestfef In yeUo'W mice^ Aa even larger difference it seen wheri the 
ftiuLmteer of hepatomas per mouse is comparedo Positive corretatio«« 
(STtiat testwees fre^uerjcy of tumors af the liver and body weight- boay 
&eigfet asTvd iength of the femora, 

U^o Mo Ko Oeringer has previously reported a reductiaa is> 
ib,@ incide^sce o>i mammary lumsrs in mice of straisv CSHe <»Mch shm 



produced by transplanting fertilized C3H ova to the pregnant uteri 
of C57 black znicso It r^ow appears tbat DBA/2e znice, coinparable 
to C3He, have even fewer breast carcinomaso 

Mice of the hairlesa strain HB develop a considerable number 
of skin tuznors but principally those arising in derznal appendages 
rather than from epiderxnis. At any rate, they exhibit a geneti" 
cally determined tendency towards turnor formationo Ability to 
enhance chemical carcinogenesis in the skin by painting with ethyl 
carbamate is well established, though urethane produces few if 
any epidermal tumors by itaelf„ Dsto Deringer has not been able to 
influence the production of skin tumors in HR mice by topical 
administration of ethyl carbamate « 



CANCERS AND THEIR PROPERTIES 



Knowledge of neoplastic diseases has profited enorsnously 
from search by iinaginative, diligent and talented scientists fox- 
common denominators that would permit developineat of unifying 
concepts relating all cancers to one anothero At; the present tizne 
everyone would probably agree that the transformstioa from normal 
to neoplastic is basically a change in cellular behavior which is 
heritable and irreversible -- and probably the general acceptance 
of the somatic mutation theory of cancer production zneans no snore 
than that. Associated with the cancerous change or consequent upon 
it are varying degrees of taxonomic abnormalities in the affected 
population, and the loss to greater or les@ extent of specialized 
cellular functiouo The normal tissue of any adult organ has certain 
definable chemical attributes which vary sonnewhat among individ- 
uals of the species. Neoplasms arising frosxi these same organs 
may reflect widely different biochemical patterns of activity, and 
when one compares tumors of different histogenesis among thezn-^ 
selves the complexity is cosnpoundedo 

Borst in Germany and Ewing in the United States emphasized 
the diversity of structural patterns and biological behaviors among 
cancers arising in the same orgauo The thesis has been accepted 
by clinicianSg and recent experience in cheixiotherapeutic excursions 



has intensified the need for better and different classifications of 
malignant neoplasms than now exists It seems desirable to re study 
the biochemistry of cancer to 

1, Delineate certain qualities measurable by simple techniques 
with a view towards correlating rather gross metabolic characteris- 
tics with biological beh&vior„ 

2o Establish the degree of change of certain specific enzyme 
activities in neoplasms arising from a given organ or tissue, and, 

So Search for progressive changes in metabolic attributes or 
enxytnic patterns as the recognizably malignant tumor progresses 
to an increasingly autonomous state <> 

A part of this is already in progresSo The orientation of much 
biocheznical activity in cancer is concerned with the description of 
drug action; yet equal concern with the natural history of selected 
cancers as definable in chemical terxns might well expedite progress 
in therapeutic research.. It is unreasonable to expect the biochemist 
to solve all the problems of cancer by himself, and» indeed, a better 
familiarity with neopla/itic diseases will require development of new 
techniques and concepts, perhaps even new disciplines of sciencSo 

The search for common denozninators should continue, but 
there is every reaso-a to focus attention on cancers of common 
origin or different Irehavior within a histogenetically homogeneous 
groupo The final result may reveal entities as discrete as mumps, 
chicken po% and m.sa8les within one anatomical type of cancero though 
one would not esepact such differences to be associated primarily with 
qualitatively diff/,-rent etiologies. 



New tumors 

The coJony of mastomys established by DXa H,. L, Stewart is 
developing a lew carcinomas of the glandular stomach as escpected 
from eyerie nee in South Africa, Hepatomas are the most frequently 
occurring neoplasms at this tinae. Attempts to transplant tumors 
within the species have been unsuccessful, 

Dwo Dur.;a is studying several unusual uterine neoplasms with 
markedly org-unoid structure which arise in strain BALB/C miceo 
One of these iiiter seven months' growth as a transplant produces a 



:» 



cystic cavity lined by erxdoroelrium wilh a w^li dgliaed mfom& 
and an o«ter layer of smooth wmacteo 

A transplantable osteogenic sarc Ki«<a «tadl#d by DySo Dus,?« 
and Daw® contain® many lasrge nvaltlawi';i«at«€6 QSt#ocia«tic c©ii@ 
wMch disappear when the lesion is gvoofn^v^ro bat reapp^at> w&e 
th© tissue culture cells are fcranapiant^/d back to jT.ice„ 

Some interesting lesions ha^e been found in Dr, Deslng^r's 
colony of strain BL mic^s. The majot-ity of the Imtn^inm dm^milp 
large o^ries due to deposition of amyloid in corpora lut®a, Tli© 
o^ries are also one site of a pecval-.ar n®crotisiiig arieritlJ wMcfe 
affects other tissue® including the hidneys, thoagfe is« ^m.ct distri- 
bution and frequency are not yet de.'ierminedo Strain Bi..,, which aiso 
deipelops some nnammary and puin.-ioaary tumors, h&» \m%'-> reported 
by Drso Leon Sokoioff CN;iAMD) as»d Ho To H^hmrmstn (DB^} to ha^e 
ideopathic necrosis of bone as wc^li. 

The Institute maintains ab';.ut i"?§ differ©t>t traRspiantabie 
tumor Ss and specisrr.ens ha^e been ®#nt to ail parts «f sh® w^rido 
Each represents special characteristics, sm4 m^^wy n@w n«opiaL-. 
that can be propagated becomes a u^minl torjh In e^omm ^s.ms Rew 
tumors generate entirely new pifograros, 8e«#arch on plasma ceii 
myeionr.as is a case in pointo 



Plaftaaa _c^U® and piagMi_aeyt^^ 

Dro Dunn described a plasma cell tu3^-.r arising in a C3H mouse 
during 1954, and maintained It in serial trans pla?^t&tt<m. Dr, Michael 
Potter in 1956 commenced an eagperiinesslai progra??^ based on araiu 
ability of Ibis isiyeioma to which others Jm^e bean add^d «ubs@^u#ntly„ 
Hi® infectious enthusiasiai focused the attentlfm of cc-a^ag,!^® on the 
•w&Lxte of eieperimentAi frsyelomas as re^sarcb toc-is., ar)« some infor- 
mation was sunfimariaed in earlier annual reportSo 

Plasma cells first appear in the ir.esenteric iyK^phatic tissue 
of mice at about the tenth day of lU^g according to Dr, S„ Co 
MacCardleo Their xtiost distinctive structural feature Is a juxta- 
nuclear, clear, cytoplasmic "hof" which contains the Golgi apparatus 
and the centrosomeo Plasma cells in the ten»day-old mouse have 
distinctly smaller Golgi than those in adult mice, E3earr.ination of 
plasma cell neoplasms leads Dro MacCardle t© speculate that the 



i 



myeionria ce-is are derived frosr. lyinplsiOisrsa»iik# ceUi? which In tur«3 
probably orlgiaafee f?oxr undiffereritiated" lynriphocytogenic €®li» i^o 
the geffmins;. tseBteys ol lymph nodeSo 

Dffo Emma Sfeeltonp seeking concrete information on ihm 

beiiavior ol t&orznal cells in the presence of tumor eel:.®, studied 
the behavior of normal cells when grown in double diffusion cb,»jy.' = 
bere placed in the peritoneal catties of sniceo Cells carsnot lra= 
verse Ifee walls of these cha.mberSp but fluids can. Ceil® of the 
peritoneal fluid placed within chambers ttltiwiately forsr.. a« organ = 
iaed tigsue containing typical fibrobia®t«o Detailed mtudf al the 
sequential events leads Bro Shelton to conclude that lymphocytes 
grown in this imanner niodulate into an indi^erent cell typiS; which 
has the capacity to becortie either a fibroblast or a piaema cell. 
Macrophages appear to ha^e the same capabilltieSo Grarsiilocyto- 
poiesis occurs in the chair.berSs and most ceils persist b«yand 140 
daySo Collagen can be identified at two weeks and its conical then 
increases with timeo 

Thus two different approaches complesnent one another insofar 
as the origin of plasma cells is concerned^ and Dro Shelton has also 
awakened a long standing contr j^^ersyo 

Substantial impetus was gi^en to plasssiacytoma research 
through the occurrence of several new plasnia cell myetonr.a® in 
mice into which diffusion chambers containing mouse sriammary 
carcinoma cells had been implanted by the late Dr, Go Ko Algire 
and Dro Kuth Merwin as described in the report for i'^58o The 
exact reason for this phenomenon is still not clear, as same few 
myelonias have now been obtained through implantation within the 
abdominal cavity of empty chambers containing no cellSo as repor- 
ted by Dro Merwino Eicperiments testing the capacity of v^arioas 
parts of the diffusion chambers to initiate the plasma cell tumors 
hs'^e not yet been completedo These myelomas have a distince ad^mn- 
tage over the earlier plasmacytomas available for study since they 
grow more rapidly, A series of individualistic plasmacytomas is 
now available which reproduces most of the features of their human 
counterpart. Including circulating myelozna proteins characterized 
by Dr„ J„ I.0 Fahey. 

Morphological studies of these ne 

correlated with biological and biochemiccti aitsrlbules at 
Ore,, Potter and, Fahey reveal ihres. types of Golgi appgv 



i\5i» sm ;;■;.■.■. att&nd»» a dibuss uevwork, or & iarge-j thick a«d 
hyperfci. uphied ball -type nctworko Ceiia ol two tumosrg which pro- 
duce p-giobuIin« hmfm the third variety, but Ih© typ« oi Goigi seeKi® 
to relate better to th« speed of production of myeiosana proteins than 
to their electrophoretic properties. 

Dr. Ao Jo Dalton has e^mrtiined uitrastructuire of 10 traneplaiitable 
pl&srria ceil neopi»8zr>Sc Di££er@nc@s within the group relate princi*' 
pally to the extent and cotxiplexity oi the ergastoplaszug but these do 
not correlate well with the physico-chesnicai characteristics of the 
myeluiTia giobuline. A feature common to all ol the ttunors is the 
presence of janinute virus ^like intracytoplasrolc bodieSo The parti- 
cieii are formed on the membranes ol the endoplasmic reticulum^ 
and the membrane itself forms the outer @helK. Similar bodies are 
absent from nornnal plasma ceil@ ol mice^ The significance of the 
particies i© not known, but they may well be related to the neoplastic 
»t*4feo They are not the mammary tunnor agent» as some of the mye = 
iornit» have arisen in agent -free mice and bioaesay has failed to repeal 
presence of that '^iruSo An attempt to es^mine cells from human plaS' 
ms^cjtaxr.^^ under the electron microscope has been beset with a geriee 
q£ technical diHicuities which are yet to be surmountedo 

Evidence that the myeloma cells secrete the myeloma prot«in@ 
into the serum was published in 1958 by DwBo Nathan^, Fahey and 
Potter,, Attention then focused on the relation of these proteins to 
th'^ss aormaiiy occurring in the serums Dro Fahey has characterised 
tbe physicochemical properties of human garatnna globulins which a'i?e 
c 'sariy a large family of related but not identical protein molecules^ 
Twa major groups with uitracentrifugal sedimentation coefficient® of 
18S and 60 6S respectively can be separated by anion -esechange cellulose 
ckgomatograpfeyo The 60 6S group can then be further subdivided into 
?. or arrofjg fractionSo Detailed characterization of these fractions 
res?ea{.« a femi.iy of molecules sharing similar uitracentrifugal and 
i?rri.j';i*si:.jehen:iical properties but differing In electric charge, hesEose 
co-aterat and antibody acti^ty^ 

A similar family of molecules is found when myeloma proteins 
t^^'m g^^ra ol patients with plasmacytomas are esamtined in detaiU 
ris-(g®© »iyeloma proteins appear to represent proteins normally present 

in eerum in ^maii quantities. Similar observations ha^e been made on 
ttn^ »®s-am rx-iy© ).on-i& proteins associated with eight tx-ansplantable 



2?„ 



Dr. Fahey argues that aince fthe variety of myelomat prctsias 
represents a variety of pLasma cell neoplaszziS, the spectrum o£ 
normal 6. 6S gaxnzna globulins probably represents a spectrusn of 
individual normal but distinctly different plastna cells. 

Immune response of myelomatous mice to sheep erythrocytes 
and bovine serum albumin has been studied by Dr, Falconer Smith, 
Capacity to form antibodies to either particulate or soluble antigens 
decreases as the tumor grows larger. The secondary response 
remains unaltered and the half -life of passively transferred anti> 
bodies does not change. The results suggest a factory so busy 
forming myeloma proteins that it gives an increasingly lower prior- 
ity to antibody production as the factory eispands. 

Dr. Michael Potter is studying the effect of chemotherapeusis 
on the transplantable myelomaSo Somie of them will regress 
completely when treated with 5-fIuorouracil even though initial 
treatment is delayed until the lesions are 2 cm. in diameter; others 
fail to respond. The optimum dose of 5-fluorouracil is 20 mg/kg 
siK times weekly. Toxicity occurring within two to three weeks 
requires 50 percent reduction in the dose, and the tuniors recur. 
Some mice apparently cured of one inyeloEna have accepted a second 
graft of the same snyeloma in contrast to Dr. Abraham Goldin*s 
experience with mice cured of leukemia LIE 10 by treatment with 
3', 5'-dichloroamethopterin (vide infra ). 

New plasmacytomas will doubtless appear in the course of 
time and be added to the fine collection now available. The progress 
nnade to date already permits an aggressive attack on specific 
problems concerned with causation and treatment, and also empha> 
sizes the usefulness of these tools in pursuing basic problems in 
protein chemistry and metabolism. All logical reasoning requires 
a considerable expansion of the chemotherapeutic effort. The indi- 
vidual tumors faithfully reproduce the manifestations of multiple 
myeloma as seisn in the clinic, including some variety of individual- 
istic behavior ])atterns. 

While plasmacytoma is rather well defined in biological and 
chemical terms, macroglobulinemia occurs also under more poorly 
understood circumstances., Dr. Fahey has studied gamma macro- 
globulinemia in patients with a variety of diseases, including the 
syndrome described by Professor Jan Waldenstrom of MalmB, 
Sweden, who visited the Institute during February, and the Bing-Neel 
syndrome. Correlative studies by Dr. T, F. Dutcher of lesions 



■LZ, 



among those patients with cancer and tnac3Poglobulinainia ha^^e 
revealed characteristic lymphocytoid piasina cells with inlranuclgar 
inclusionSo Dr§o Dutcher and Fahey suggest that the intranuclear 
periodic ^acid-SMff posit4'i<*e material and intramjclear vaciaole® 

seen on phase contrast rriicroseopy ar© chemically identical to the 
circttiating iSs, he}eose=rich Y"=f^acroglobulino Sequential cellular 
changes anioag the Ifmphocytoid plasma c©Us suggest tho8« cells 
as the sit© of rr,acrogU>feulin lorm&tion. The investigators regard 
Biacrogio&alineixiia as a n^oplasjrn of reticuio#ttdath©iial origin^ a 
¥-ariant a£ rniyiltlpie rr'yeiarna or a ^pe of lyiinpfecrriao 

Biood vlscaslty iXi&f be markedly gl^vated in inacrogiobu-lln-irnir 
and was the imfinfediate cause of cardiac failure in cm® of Dr„ Fahef "s 
patientSo H© has d.e@lgiigd a th#rap@atic regitngn lor maintaining 
th© blood viscosity feelo*' ihm critical level. 



pi@©*rs\isiati©0 of '-^r&ncer 

. c,m ability af cancgrum* celia to ir!H*-ad© s^arrouading lissufes 
and coiomse dislant parts of the body through access to ^ascisiftr 
cfeannets i# th® n.^'St Irrportant factor Hiriting th® «lfectiy#neaa oi 
.^'arglc&A :.r raditHt^gicai sherap-^uaiSc. While lh@ roales of dissen- 
ipAtiofe ar% rgaaoRably w-#U u!ad#r©toodj th« ba«ic blolagicai lf><' ts) 
re©poiasl&ig for :a etaetaais art ubscursj eepgcially sines ff-.^-tcs?-? - 
siaaMlity .'.(ay tee at\ attribute acqair^d after th@ tr&n®!- :' 

,-1, ,ja-«i!.£Qn *s =sSv*ci|'t'.i| @xpe-rus<ental lyrriphoiti vts ■■ - 
K'Mck rgifiK&lft ioeailzed on transplantation or dissejr.tnet 
rapidly„ .-'Ifferenc^ In tim cgnaeities of thgs© n€upla*iXi^ to ^-zin^i^ 
trate the *s^U® %if dyfusidfi chamber® has bgitn descrifegd m ga^ii^r 
fepo'i=ts<. :,.jr?r.ai csUs of the peritoneum and tnv/asw© ',yjnphorn.a,s 

Acl^ or ■ ^iO* itidgp«ndently of on® another when ••^i.9.c9.6- It? 
atfftsgioB «:fes?:.fi}'&@rfiio The growth of ®uch non^in-^fast^^ vympi- 
cells a® I.,' A .d P3§3» however, tetsds to b@ aasoeiatec wltb 
of fe© payii .••■^-gai cells, aince the fe&mor c©Us grow in <iMRm'^ 

'i-\Mngt% ;, .su.^-;- anvleo strni enclosed by normal ce; 
;fif,--.r,- sufe., v.ijs..-=,^;^(a,,g fat L*ad© i.i Ih© j.yto?sge grow -^ 

-I. :i:-*rriib,grg„ and Or.- Sha.'iton has i?JtrodiiiC#d n#optaaii4c - 
.-■t «,3i0 fibro'fl'iaat pupulatioHa Agaln^ cells frot>, ; i r n ^j : 
Ireis-^floatlng ItftdepgRuent eruifttg®., Gro«fih of ' ,. ^ ., 

feiafed is pr«>lifgratiBg fibrobi&stSs and th«s© feu.','.. ^ ■ 
fS'CfW beyond tha ad'^'^ncisg gd;|'i of tb-i fibroblast solc'rjT 



i.;^ ;. _: jint effort of the Surgejn^; Branchg the JLaboratory of P'■ 
a'ad ibg PHS Hospital in Baltimore^ led by Djfs,. R, R., Smith; ... 
Malmgren and J, F.., Potter. Cancer cells are found more often In 

the efferent blood from a rriaiignant neoplasm than in blood take?: 
from the antecubital veitio TMs was to hB.ve been ©sspected and c-: . 
relate almost solely to dilution. ©ICectSo though the filtering actiori of 
the Iwer, lungs and other organs cannot be e3«.clud@d at this Hnie: 
Patients who&e cancer© are appa.rently amenable to definiti^- . 
nrient have tumor cell® in perip.hej'al blood abouf- ? ■■' r'"^ c':*'- -; 
those whose disease is recog»?zabif difisenainaf 

Circulating tumor ceJ. oeen found in peripheral venoi.«s 

blood in about 75 percent of patients with inalignant mslanoma.., The 
cell© are seen sporadicallf In serial specimens gf-en though the 
nrielanoma i© widespi'eado Only two of six patients whose melanomas 
wer© surgically resectable had recognizable cells in the peripheral 
blood preoperativeljo Both of thenn rapidly developed rectirrent 
dsseage and died 

The significance of circulating tumor ceils will become apparen? 
■'n.ly aftes' a Ij-ng comprehenahf'e 8t@dy of specific anatomical types 
of cancer in lndi'«idual patients „ Correlated aninrial studies are in 
progress^ but the sanne methods by which one prepares specimens 
of human blood cannot be applied to mouse blood lor som© obscure 
reasutie and„ of course, > the gmall siz© of most laboratory animals 
that d@¥eiop cancer in a reasonably p,r*5d4ctable fa»Mon or will 
support the growth of 'transplanted nsoplastxis militates against the 

of serial aipec.iT7i©ns from tKp= .9.:»rn,p inrlividui*!,. 



•- 'ther studies of metastasis -m ej;pt:^.«.-rae(:. ?_&.'. <?:nimal8 have he-^n 
especially rewardiu.go Housing tunrior bearing n'iice individually 
has produced a ma.Tked reduction in the variability of results as far 
as metastases studies are concerned,. The ability to identify and 
count n inute deposits of non-pigm-ented tumior cells in the lungs 
has been greatly improved by Miss Hilda Wexler, who introduces 
a solution of india inx into the trachea^, thereby increasing the con- 
trast between nori'na..l and cancerous ' ' ' i ' 

A iriiixh terggr «?sperienc@ in fehe situdy of pulmonary metastasej 
prod'ttced from transplanted tumors growing in the thighs of gene- 
iicaiiy appF.;.priate miU.e points to a uniform type of behavior among 
the n©opiasniS ini!'esti^i;a'.ted<, Dr., A, S, Ketcharn reports definite 



sfes'aits obtained by Ox 



too! in imc &<t stiid-r 

have been obtainec 

piasrTi by trocar tra.aF;; 

to b.a.i''e been rathex' in:' 

it necessary to trauspl 

ujr^a Iferough 7 to 9 se; 

rr>etastasi@ develops wr 

fence xnAf be ejctremely ^iT-portant ss;:--^- -l su<j:/ 

oi aeie-cfeioia may be involved in the proc'.uctio'rt ^ 

asid that orsly sonrse of the cellular pop^dfttion oi 

piasii rsr.?: capable of n'et^a.staslamgc This phsv 

be s.-jgS-. 

iu^qioaiiy fascinating is an obsr 

Jhe iocaiisatiorv of n-ietastases as ^epci-.~2 (•;;); Ln-.-. u, i-^.- Ji't: 
ia&t fear''!S report referred to the predilectioii of tte luAga^ . 
of mmiMnotnSi S-9i iratroduc^d into either arteries or \-'ein® despf'irs 
tfee ability- of the sasne ce?lf- co gx'o\w wfeen mtroduced directly im'/. 
%^ariu^a§i organs and tig-'' Metastases can be produced in tlie 

iwe3f wh^R large^,. appr-v. j. -.. . ■.■.«iy lethal ' ■""'" ""' ~ '^'' ■-'■''"■ ''"' 
iat's-C'<avi.cnci into feributaries of the porte. 

planted pieces of liangs to the thighs of c,s.:.'3-::£caily suitable n.ice : 
obser^ea groAwtK of nneianon-ia ceUf Ir. i-oth <-h^ ectopic aad ?iorr^ .:. 
pui■a■icma.rY tissue when tbA-' Imtc rl into the blooo, 
stream. Transplants o£ other -_,,,- ■< j-fee- thigh would 
not support growtb, of inlTavascuiar .. ,eiano£3aa 
v5~';l. Tfcis seems to bs the first exoe • ; ,i-;i<fc.i cun,Mi-M.o!.fcion o£ Sche 
febesis a.®cribing certain bszarre patterns o£ trietastasts to a locus 
miaoris grtfei3tentiae„ In tMs particular example the feasic mechan- 
i®n.i sjay well r^lT,:te to the knowtj, capacity o£ the Ivsaga to fUter cells 
fro/n the '-ig'culat'iing bloodo 

iJr,, -!* h jHu'. ";■:'■- "' : * ' iJae the growth o£ a 

ha'\fe not toeeu ^iiccessfulo 



Experiments conceyned with cii©iriotherapeutic pre'^'-ention of 
metastasis formation or treatmertt of artifically produced puirrion- 
ary metastases have produced no significant new findmgSo On the 
other hand„ Dvo Ro C^ Hoye finds that treatment of three out of 
four different experimental mouse tumors with 7i5r X -irradiation 
reduces the incidence of pulmonary metastases by 70 percent and a 
dose of ZOOOr effects 81-100 percent reduction„ 



P olygaccharide research 

A long-term interest in polysaccharides at th© National Cane?' t 
Institute has stemmed from the observation by Dr.- Mo Jo Shear aud 
his colleagues that this class of macromolecuie can daxTiage both 
clinical and experinnentai neopiasm^r. Certain problems arising 
repeatedly over the years required some esepansion of this research 
area disring 1956„ designed especially to broaden its scope,, especialbj 
at a fundamental level » Substantial inforyrjation has been obtained 
regarding many important aspects of lipopolysaccharideS;, 

Bacterial endotosdns are important members of this chemical 
specieSo During the year Dr. Maurice Landy and Dr.. Edgar Hibi 
(NIAID) fea^e reduced the lipid content of lipopolysaccharides from 
19c. ^ to l.,Z percent without reduction in their tosricity and without 
detectable effect on endotojcic properties, Dr, Woods reports that 
bacterial endotoxins in concentrations of Oo 003 to 0.. 3 p^. p.mo st4mu= 
late aerobic glycolysis of cells from melanoma S-9i,, and correlates 
the degree of stiinuiation with the degree of acti\d.ty in 'ifiyOs as 
determined by Dro Landy <, 

The «ndoto3£in deto%ifying component (£DC) disco%-ered by 
Dfo LandY; which inactivates polysaccharides with respect to tumor 
necrosis and stimulation of specific antibody formation,, also acts in 
the same way against killed So typhosa but not against the viable 
organisms,. The latter can b@ inactivated by sera from several 
©pedes by a different and bactericidal system.., Norm.al antibody^ 
ail 4 recogaiized compots,ents of complement and dii:?alent cations 
ar© required for the bactericidal systemSc. The concomitant elim = 
inatioft of eadotoxic attributes of typhoid bacilli in the bactericidal 
process of ftormai serum has obvious important innplications for 
the host-parasite relationship^ 



;-• .isrna of febrile rabbita tlia 
UpopoLy saccharides has been reporiad ;o Cont<ii;-.n ■ eudagea^^us 
pyrogen, '• a substance with fever producing properties quite dii: 
froHi those of the polysaccharide injected originally. Since rec 
in the pyrogenicity of endotoxin followin---; inr-uirit-. r,-' in. virro -c' 
norn-al hunian serum was reported ye; - 

a study to corr.pare the properties n-'' '' .. ogeir 

endotoKins incubated with serum. ties \ise-r:. 

distinguish "endogenous pyrogen" ircm i^oiyiiiicciuiride are : 
which appear after incubation of the macsromolecules in ser - 
respect to; 

io Fever production in endo' 

Z^ DissociatioT- of leukopenia 

3o Lofia of tolerance to dai'- 

-. .. ."„;-. ure to develop acu.j; ; 
injections given over a sb 

Strains of E<, coli yary in the. 
systevn in serurrio Study of tvi?o wideiy diverge "at stireiias by Dr. 
and Dto J„ C Michael (Visiting Scientist) reveals that the cone 
tration. of polysaccharide 4s inversely related to susceptibility oi 
the organisin to the bactericidal systenic Cells =r-^^ o,-v ;-.••<-. o? ' 
resistant strain esdiibit greater endotoxic poter 
ability to necrose sarcorrja S37o Normal hums-n ana 7' a ::■:;,•:■•■ 3i\: : 
contain lov.? concentrations of natural antibodies specific for bsc 
of several genera of the Enterobacteriaceaeo Their he : 
action is not increased for E„ coli by addition of a spec 
antibody^ 

.rtsse obser^fatlons haife led. Dro hB.v 
rm normal serum which affect the viability o.t tumc'^ 
of cells from sarcoma S37 with tsornnai human sert 
capacity to grow in xnlce,, Metabolic activity is lost., as m- 
.In Warburg flasks by Dr„ Woods., 7fhe cells no longer redv? . 
-trazoHum„ They stain readily wstfe eositij and blebbing of the 
.nembranes occurSo In short., they are dead„ The effect of ■- 
■cidal action can be eliminated by incubation at 52*^Co for ,^ 
adsorption with waSi'" 
ored by coiribining ' 



is deci : '?%© milliiite V 

million c^^ii- •*■ ..r-.- •--v ^'^o^ ^^^ ^^ wias- 

spread in natuv-: ;>eci@Ss, mor 

dilfe^f?- ■::cr, but 

the onl; 

loerido'-' 

■klT3.nd fState Baeteriolof- rjkfeolm, 

it cat? ■; ~ , pure 

' ■ ' ■ o-blished 



34o 



DTo Ho A„ Saroff CNIAM.D)s reveals a pKa of about Z„ ?, n ost likely 
due to carboseyi groups of the Neuramirac acid typeo The great butk 
of the polysaccharide is a polymer of glucose contamirjg 0, 3 percent 
phosphoi-us in some organically bound form and an extren-^eiy smaii 
amouiit of amino acids,. 

The ability to bind certain molecules to synthetic substituted 
polyglucoses by electrostatic forces and release the bound Riolecuie 
by adding a second macromolecule with a stronger charge seems to 
apply to many dilfereni ensymes^ For exarxipls, the most ift^portarj; 
factor in inhibiting ribonuciease acti\4ty with the nrsacromolecwies of 
interest is the charge density as mea.sured by degre© of suliation.„ 
The maydrrmm sulfation is 3 per anhydroglucose unit which prodace® 
more than 1000 times the inhibition possible with heparin, the best 
agent pre^^iously descx"ibed possessing similar acti^^'ity^ Charge 
density seems to be the most important factor in enzyme inhibition 
also with, carbojcyl derivatives of polyglucoseo The molecular 
weight of the inhibiting substance must- be at least 2000,. but th© 
internal con£igu:i°ation of the molecule appears to be less iinporta.rit,; 

Other physicochemical studies carried out by Dr^ Mora ha,v<?? 
p,rovided further information on ensyrne-substra-te interaction 
invoi'ving two oppositely charged macroitioleculeSo Titration of 
polycations %vilh polyanions £oi.lows stoichiometric equivalenceo 
Such Interactions differ .fronn those involving low rr;olecular weighs 
ionSs since a firm electrostatic complex? forms under certain con- ' 
ditions., v/hich leads to precipitation of the ma.cromolecular com- 
ponentSo The precipitatioxi phenomenon can be used to fractionate 
n-jacronn.olecular substances and the principle has heen used in con- 
centrating EDC activity from human serum by Dr,. Mora and Mifo B, G 
Youngo 

Administration of polymyxin^, neomycins strepton^ycin or 
protamine to mice in doses which cause death within 25 to 40 minutes 
can be counteracted by a neutralising equivalent of anionic synthetic 
polysaccharide dex-ivative administered by the same intrapeyitore - '• 
route if given 10 minutes after the toxic dose of the cationic ant- 
a-j :re::'o--:3d by Dr<. Mora, M^o Young and Dr. Sheax*,, Prior ^ 
■•■'■' 'Section of th? a'y?.tid.ot'? 'OT^''-~'i'i'>iB d^ath 'fTOTVi irw 'jc?^ ' 



U-j pox-tant biological activit-t^s are e.sefefgmei^'' inpujetaftit fu;e offiiy ir- 
- t-u::i.f/on to the natui"© and R."iechanism of oon^spgcific ffeaislanxe- 
■1 as irriiViUtxoIogic factors concesfr.ed with cbxacbs,, but aisu 

;.uaclxvati;- dose a.^'ft-.easured t 

ibitity of t.i.i,. --....,.-_.,.-.„... „.. ;. ..- vacuc iatiafs in tatT('.<r 

16 Ihi es tte doj 




. l4r>.>o.4:(,f.;g,/apaji<. sep^ratjan ..'I so;able proteins as i'u 
, Ao Sobsr and £e A, Peterson has had a pf i.:..C;->aftd. aft. 
I Uitf di5velopir<ent of bLocb.en istry throughout ttee worid,;, Isois?: 
antibodies from serum is re-ci.iilv «ccotr!plishied by a ^AcitAf^ r 
'oce-dure described by tix Dr, Peterson s? 

•lAlD) which s'ecy^ "="=<■-■ -* •■^'■■-.'nt of thrf gar 

macrogJohaiiiTS , j cars be obtv 



Chs'oinatygraphy is also being aiS«d in c«Uabojratior» with 
:% B, i..o Vailee (Pei^'./ BenS l^^l*s,a, JIo5.:<^i.ai, BocK^^^;. u> i 
-efeal-cotjtaining p 



,^;.f:c^ i'ctessoTi axiii So'ser caa produce eKt8ass.v'et,y' emBoWee 
'iacible ch.ron?;toe,rarsTS of serum or plasrxia in sijt hours, 
■raitaiion of llov," ^ "'ydrodynajrdcs but resalutiorj is not g. 

!(icxc6 bv Ihc h'-^.- raie?; D'srxnitted by the tnechamcal p 

'"fey e^chiirige on Sephade^. p.; 
- • "-' oi equilit " • - ^ - "c' ; 



The principal problem created by pjfotein. chrc P- 

Is the considerable variety of physicochemicaliy dis<an t -jtti&ies 
with the same or similar biological or inn rrmno logical properties 
(See Annual Report 1958)o Drs, Peterson and Sober report- an 
increase in the heterogeneity of iSerurr; proteins when treated --'sth 
8 M urea as measured by electrophoretic analysis of the ci 
to graphically separated peaks. The urea treatrrient, howev 
not alter the chromatographic elution profile significantly o 
-ludy of corr!ple:€ing of human serunrj mercaptalbumin with rtpidt 
.reveals a markedly increased affinity of the protein £or DiSAE -cellu- 
lose. Addition of a cationic lipid, hovt/ever; can abolish affinity of 
the protein cornplei^ fcr ;;TnB adsorbent but increase'?, ite adso?^-ji^c- 
■o CM -cellulose, . • ct the lipid -prote 

Xrea.tment o. lutions of an amino :- 

aldehyde in the preaence oi copper ions as described by DrSc To T„ 
Otani, Nobuo Isumiya (Visiting Scientist), S, M, Birnbaum and 
Milton WinitE proceeds with the formation of the coj-responding 
a-amino~p~hydy;cx-' acid The fundamental reaction ha? parir-i:r.- •; 
iingle step pre 

1 P-hydro3Ey-p-methylaspai=tic acid froiri pyru'«fic acid and glycin? 

Z„ p-hydx'OKyaspartic acid from glyoir/lic acid and glycm 

o-nvdr o-.iM'leixcins from iaobvityraldebvda .sad s^iv?c-';ri.e 

'. , ...i&ijyae cMiu giyciU;." 

E%:idently vhs nje-thcd is a general one providing asirriple and ecoaorr;- 
leal route to the pi'cduction of a-araino-p-hydxoxy amine acids use.fwl 
■jii b;och.: niical res- 

Dx , Ms.,;: CO Habiv. the inbib 

synthesis by aramo acyo. a-iiaioas a.-.u t-tuaied severai :.u vo- 
methylene group in the side chain has been replaced by Puli'-J 
Methionine suHoximine reduces by 50 percent the rate ox pr ~ 

SYiithe.T-lr :n v/ashed I-~brlicb s.rciteB ca'.ln throti'jh pre-.-^rtio^ 



DTo K„ Michi (Visiting Scientist) with. DtBo Bi^nbaam and 
Winitz Jmve separated diastereomeric n-iacfc\i'.r©s of an-iino acids Ssucit 
as isoieucine~aiioisoieucine„ hydroseyp^oiiTae -al].ohydro^.Tp"?oline 
and threonine -allothreonine by chroxnatography on Amber Jit© columns 
in amounts as great as 20 grarr^So Recovery is 90-95 percent,, Tte: 
prograin initiated by the late Dro Greenstein for the production of. 
optically ~activej puj.=«: amino acids in adequate amowint anfe at reason- 
able cost to provide yiowes'ful tools for rosearcb in the chemistry and 
mefcaboiism oi .cids and proteins has been completely succe" 

Dr^ Winitz and Dro Birnbauin with Mto Mo G, Otey and 
DrSo To Sugtinura a,nd V„ Mitbaiider (Visiting Sclentjsial ha/tre 
commenced to use chemically defined diets to st:^^dy specific enzymes 
and metabolic processes in the infcact rato Earlier worker© having 
shown that the D -isoiners of the essential amino acids rriethiomne. 
tryptophan and leucine can replace^ in whole or in part^ I.— isomer & 
of the same compounds in rats, the subject was restudied with the 
chemica.Uy de£ined rations <, Methionine \ivas the only one of tha^ group 
in -which >weight increment in vweanling rats was equally good on D 
and L isomerSo D -tryptophan, was only partially eMecthpQ in. repiac^ 
ing its 1^ antipoda and i„4 times the amount of D -leucine was re 
before any growth occurredo A series of diets was designed to 
disci'iirinate betv-een Lhe relati^'-e capacities o£ D -amino acid, oaeidaso: 
to conv'ert the .D -arrjino acids to corresponding a-keto acids and ojf 
transaminases to accompJish the transforrriation of a-keto acids to 
corresponding L -airiino acidSc Ail possible combinations of the 
isomers of alanine and methionine weife employed, but only the diet 
using D loTvns of both ammo acids produced marked retardation in 

VV P \ o '' .' '< '■ t"> c ■'^' <■:' ■' T' '=? 'At - 

Oinerr ^nva avsgators had shov^n sodixinri benzoate to be a potent 
■r.Mbitor of D -a.mino a.cid osiidase activity in t rit.ro » Synthetic djeta 
containing 2 percent scdiurn bensoate and D -methionine produced 
itrikingiy diminished rates of weight gain^ according to Dv\ Winlts 
ad associates.. These data point to oxidative deamination as rate 
■^■i'-nig., and s-\"^^ = '^ on the effects of the a-keto analog of methi-"^' 
progrer additioiial point of interest is the spajrin::- 

:;;.;ct of dietary g;i;.'cine on the inhibition of D -amino 5:. 
.odium benzoate through its removal as hippui'fc acid; 



38c 



Branches in which Dr„ P, „ Bo Couch has served as the co-ordinator„ 
The earlier diets were aemi- synthetic and contained raceroic mix- 
tures of amino acida as the only source of nitrogen^ and the general 
coinposition conformed to recommendations published by the Food 
and Nutrition Boeird of the National Research Council insofar as they 
were available„ Substantial progress vjsia made, bowevei*, from the 
experience obtained by nutritional studies in rats by Drs= Greenstein, 
Winita, Birnbaum and Mr, Otey„ I'he source of carbohydrate gave 
considerable difficulty be cause ©tee a sive sv/eetness produced nauseSo 
Dextrose finally proved to be the best carbohydrate and, in fact, it 
is advisable to add a flavoring ixiateriai if a high deicfcrose diet is to 
be taken by mouth,, The pyoblena of frequent srr.all watery stools 
with any of the soluble diets was solved by addition of carboieyjnnethyl- 
cellulosco The. best diet, which contains L ratherc thai,! D -amino acids „ 
can be stored in dry form and dissolved as needed; fat soluble vitamins 
and ethyl lineolate mixed with polysorbate 80 to forrri a reasonably 
stable emulsion are added a.nd mixedo 

Se"</en patients have received 1?. separate trails on the chemi- 
cally defined diets, five by indv/elling esophagoatoiny tubes, and 
two orally, After it had become possible to feed a diet for 7 to 12 
days without weight loss in an asyrr.ptonnatic individual whose weight 
had been stable for some time, the critically iniportant exper'izr.ents 
were done with Di*'So D» Mo Watkin and L„ E„ Rosenberg on P^o . 
patients under complete metabolic controL These studies revealed 
a need for approximately 25 percent increase in caloric intake over 
tliat pro^'ided by an isonitrogenous diet of natural foodstuffs in order 
to miaintain vt^eight and perxriit nitrogen retention after the period of 
nitrogen loss induced by a nitrogen free diet. The experience in 
patients coincides with the findings of Dr, Birnbaum that the best 
cheiriically defined diet is inferior to diets made of whole foods and 
supports the contention of Rose that diets containing only pure amino 
acids as the source of nitrogen require a greater caloric intake for 
efficiency comparable to isonitrogenous diets of whole foods„ 



Nucleic acid research 

Compiesc interrelationships among the pvu'ines, pyrimidines 
and cheinicai intermediates involved in the production of polynucleo- 
tides are slowly being definedj and the contributions by scientists 
of the National Cancer Institute are steadily increasing., Brs , 



39o 



Schneider and Jean Rotherham^ continuing their studies of 
deoKyribosidic compounds, have defined factors concerned with their 
urinary excretion.. Dsoxycytidine is the major deosyribosidic com- 
pound found in the urine of ratsj deojcy uridine and ar-i unidentified com- 
pound are also present. These materials are not derived from dietary 
sources, since neither fasting nor the use of chemically defined diets, 
through the cooperation of Dtc V/inits, has much influence on the 
level of e:!Scretion„ The germ free state also has no pronounced effect; 
hence bacterial syrithesia in the gastrointestinal tract cannot be a 
major factoi' in their productiono Excretion of urinary deoxyribo" 
sidic compounds usually declines for two or three days following 
partial hepatectoniy and also decreases during the most rapid growth 
phase of the Novikoff hepatoma » These two response Sj however; are 
not entirely constants 

Incubation of hepatoma cells with isotopically labeled 
deojrycytidine results in progressive labeling of deoxyribonucleic 
acido Dro Schneider also finds an amount of radioactivity in the 
nucleotide fraction sufficient to suggest the presence of 14 or more 
deoxy nucleotides of v/hich some contain deoKycytidineo Admiinistra- 
tion of labeled deoxycytidine to the ra.t results in uniform distribu- 
tion o£ C thruughout the bodyj the amount of radioactivity in any 
given tissue is so low as to discourage attempts at isolation of the 
labeled compounds, Dr, J« H, Weisburger has collaborated ';vicli 
Dro Schneider in the synthesis of phosphoethanolamine-1, 2-C^ . 
and synthesis of the phosphochoiine analog is in progress, Deoxy- 
cytidine containing analogs of these compounds were isolated from 
the Novikoff hepatom.a„ and a study of their function seems desirable, 

Dr, R, K„ Kieliey has encountered difficult technical problems 
in her studies of enzymes involved in synthesis of deoxyribonucleic 
acids by normal and neoplastic cells. Older techniques for isolation 
of nuclei have required substantial inodification for use with certain 
hepatoma cellSj, Separation of diesterase and nucleotidase acti\;-ities 
in snake venom has been accomplished. 

Reports from several sources of rrjajoi' quantitative changes 
in urinary excretion of purines and pyrim.idines during chemothera- 
peutic treatment of acute leiikemia encouraged a collaborative pro- 
gram led by Dr, J, C, Beid of the Laboratory of Physiology with 
colleagues in the General Medicine Branch to isolate and identify 
purines, pyrimidines and intermediates presumably associa; 
nuclp;ic a.cid synthesis and/or degradationo Chroxnatograohic 



40» 



procedures have been used e:!£tensivel5' by Dt^ Reid in de^y'ising 
analytical methods for detection and quantification o£ i-ucleotides, 
nucleosides, pxirines and pyrimidines in human urineo Most 
difficulties have been obviated during the past two years, but 
xanthosine recovery is still low and the ammonium acetate gra- 
dient used for elution of the major nucleoside fraction does not 
yet gire good reproducibilit^r. Quantification of all individual 
componanta at the present time is an a.rduous and complicated task 
that would seriously limit the usefulness of the inethod for study of 
diseaseo Dr<, A,, Wo Pratt is working with Dr, Reid to automate 
the process to the fullest possible extent. One approach now being 
pxirsued is a computational analysis of inulticomponent spectra 
applying several mathennatical techniques made practical by the 
digital computer. 

Methods for isolation of nucleic acids and the deteramination 
of their chemiical and physical properties are improving. These 
large molecules are sometimes rather fragile, as are proteinSs, 
and corriparatively smeill insults during isolation may denature the 
inolecule, Dr„ Joseph Shs-ck finds the binding of deojfyribonucleic 
acids by n:iagnesium ions essentially Independent of pH in the range 
where -ao titration of nucleate occurs,, The quantity of m-agnesium 
bound is proportional to the concentration of deoxyribonucleic acid 
at a gi\"en concentration of magneaiurri ions. 

Evidence presented last year by Dr, Shack and Dr, Kilham 
(DBS) suggested that the transformiing agent prepared from rabbit 
myxoma virus covitained deo:K:yribonucleic acid, Attemipts to isolate 
an active deproteinized nucleic acid by the phenol procedures have 
been unsuccessful. The transforming factor prepared from myxoma 
virus by heat alone is acti%'e in such various tissue cultures as 
squirrel and monkey kidney, rat embryo and ceiia from the kidneys 
of either cottontail or domestic r?.bbits. It can enter the cell in the 
absence of fibroma, virus where its activity can be detected for as 
long as three days. Transforming factor prepared by a procedure 
using both heat and urea is active only in tissue cultures of rabbit 
kidney, the natural host of both fibroma and myxoma viruseSs, but 
this preparation is destroyed in the cell within three hours of its entryo 

Fractionation of the Novikoff hepatoma by Dr, E, ,L, Kuff 
using gentler physical procedures in conjunction with electroii 
-r Icroscope stu.die3 by Dr„ JR, F„ Zeigel has pertnitted isolation 



41< 



ribonucleoprotein in that neoplasirio The spherical particles occur 
in several size groups, the most abundant of which measures 24 m\i 
in diameteTgand have a molecular >weight of 4, 5 miIlion„ Addition 
of adenosinetriphosphate in concentrations comparable to those 
found in tissues causes dissociation of the particles into smaller 
subunitSo Adenosinediphosphate is less effectiij^e and treatment 
with deoxycholate, the detergent usually used for isolating ribo- 
nucleoprotein particles, remoi'es one-third of the protein without 
changing the spherical forrrio BXo Kuff suggests that the physical 
form ol ribonucleoprotein 2r.ay relate to the metabolic state of the 
cello 

Dxo Mc Eo Maver has previously described the hydrolysis of 
cyclic adenylic acid to 2' adenylic acid by a ribonuclease prepara- 
tion obtained from spleen„ This type of activity has been separated 
by chromatography from acid ribonticlease activityo The ribonuclease 
activities of rat lymphosarcoma closely resemble those of calf spleen^ 
The acid ribonuclease s of rat liver and hepatoma hydrolyze only 
cyclic pyrimidine nucleotides to give the 3' nucleotides. The corre- 
sponding acid ribonuclease s obtained from spleen or lymphosarcoma 
hydrolyze both purine and pyrimidine cyclic nucleotides to yield 2' 
derivatives before purification by chroroatograj 



Dto Maver also describes some differences in deoxyribonucleases 
isolated by the same procedure irorxi different tissues^ Those obtained 
from calf spleen and liver have a sharp pH optimum at 4= 8^ whereas 
the deojcyribonuclease from rat lymphosarcoma acts equally well from 
pH 4 to pH 5„ 8o 

Trypsin destroys a substance obtained from human leucocytes 
which inhibits the activity of deoKyribonuclease I but does not affect 
the enzyme. Destructive action of trypsin is prevented by soybean 
trypsin inhibitor which does not prevent destruction of the inhibitor 
by a factor present in normal human serum„ These findings have 
provided Dr, Shack a basis for assaying the activity of both deoxy- 
ribonuclease I and the inhibitor and to show that normal hun:2an urine 
contains large quantities of the enzyme but iio soluble inhibitor, 

Dr„ E„ Po Anderson, v/orking with Dr, Lo A„ Heppel (NIAMD) 
has studied a new phosphodiesterase, which hydrolyzes certain 5'- 
phosphate -ended ribopoiynucleotides to inononucleoside-S' -phosphates. 
Individual nucleotides are released serially from the end of the poly- 
mer. The enzyme is active against rlbo -oligonucleotides but not 



against sirriple esters oi rxJcieosidci-D" -pi-tospJsates^ b^jcausa of a 
need for a nucleoside: i-noisty on each, side of the siiscerplible phos = 
phodiesfeer boiid„ Enjiynies wi,t.b Himiiar laydrolytic acilv-lty ar© 
found in tvjnnor ?„ 

Drs„ „io„., .....^ ..■ ..„„. „ :,^,.. cu.... ^w., „ ..,..^=,....;.as Staebelin (Visiting 
ScleBtist) report that oligoRucl©otidee. prepared by dij^estion oi 
ribonucleic acids with paficreatic ribonuclaaae can b?; fractionated 
on ceiiuiose anion adsorbents using volatile salts for elu.tion„ 
FractionaiioTi is baaed priinariiy on diffei'en.ces in size of the 
molecules, but the ordei- of esnergence from tbg coiuran also relates 
£o basic compositiora and sequence.. Whereas oth©r r/^etfeods pern:-it 
separation of tetra- or pentanucleotides,; column cbromatography 

permits fractionation, of decanucleo'tides and even larger rr.olectiiieSj 
eacouraging ixivtheiX study of specif icitf o£ intesactiona of nucleic 
acid© and enzymes,. 



Sndocrmoiogy 

'i;^i--:n- , e metabolic studies conducted b;, . rgensta). 

and M,, Bo Lipaett with hypophyseal growth horrnoui; i.!:oi'n bei^i, whale 
and sheep treated with chym.otrf psin have revealed no chai*acterisfic 
activity in man,. Purified aiid untreated growth horirione from these 
species is simiiaz'rf ia.:2.G;.;. v-p,. Human growth horn-; one itself is a 
potent anabolic age ases the daily esccretion of wrinary 

Admimslrailov; oi gicwtii hormorie to three patients with acror;; ■ 
reported by Drs„ Bei'genaSal and. Lipsetts caused no retention oi 
mtrogen^ though the SKcretion of calcium in the urine increasec',^ 
Testosterone d.^d n.-orlucp a;- ^nrihoUc effect in the aains nrripn; • 
different mech ' td with nitro 



BeigSiiStal ^ud ieaciibe signiiican 

-■ -. ; . sxTiall dopes o.f c : -.6 tri- iodothrronir: 

th panhypopitultc. 



■ g any correlation o£ extent or severity oi 

:.aD.s.iiLy Oi p.f-cge£i.ai,iOaci.i coinpouads free of c ■ 
act on oral admimslration has permitted e?tar. 
:- ; , :.is of large doses of progestin by Dr&o Bc;rgcnstal and 
f-.:.li,. Provera (6 -methyl IT-acetoxyprogesterone), in cor-'-- - 
ogesterone, has caused negative nitrogen balance only one 
\rialSo Provera has no ant i aldosterone activity, nor does it. .'Jappr-f -' 
gonadotyopin excretion in postmenopausal patients or in girls with 
precocious puberty„ Administration of this progestogen causes ■ ;: 
increases in the estcretion of Porter -Silber chroinogensj hence th 
urinary meiaboiifces 'we.ve partially characterized. Known functj': 
groups concerned with the Porter-Silbe- ---'->'~:^'---> U:...-,. been rule 
as a source of the increased chromoger - finding? 

suggest a previously unrecognized phenyiir-cl'-a?iiis- . group 

among metabolite a of Provera„ 

OptiiTiai progestational response of the endom;- 
requires "priming" with estrogeno Hisaw, et aL , howe-^feri, repc 
that large doses of progesterone alone can induce proliferation m --'>■■■ 
rabbit endometriumo This conceivably could be related to production 
of estrogenic substance by the adrenal cortex^, Dr^ Wo Wo TuUner 
has observed this progestational response Tn ?drenalecfeomlsed rabbi-s 
thus eliminatij'ig the necessity of an 

Dr= Herts rinds that prepubertal growth and differentiation of tht 
ovary proceed independently of pituitary stimuli,, because growth and 
differentiation progress normally when ovarian grafts are transplanted 
to hypophysectoniized rats of either 8eK„ Extensire destruction of the 
hypothalamus fails to influence the process. Ectopic pituitary glands 
continue to produce ACTH<, growth hormone and gonadotropiUo 

Clinical eseperience in inhibiting the activity of the adrenal cort e: 
with ampbenone or with o„ p^'-DDD has pointed to the need for better 
drugs with similar primary effects but fewer undesirable reactions, 
AdmimstratioTs of o,p'' -DDD to dogs not only suppresses excretion of 
adrenal hormones or their urinary metabolites but will actually 



44 



destroy the adrenal cortein in 90 days.. While DtI TuUner has not- 
observed evidence of regeneratic-n of the cortical tissue, the period 
of obser^fation has not yet been adequately long to be decieive„ 
Dsapite this drastic effect in the dogj and evidence for comparable 
qualitative effects in some human situations^ o^p'-DDD does nofe 
influence adrenal cortical activity in either the rat or the monkey,. 

Studies of the pb„; ODD have been hampered 

by its incomplete adsorpdoB on oral administration^ so Dr„ TuUner 
and Dro Hertz have developed a suitable v^ebicle for intravenous 
injection of this fat-soluble compoundo Secretion of cortical l?- 
hydrojcy steroids decreases max'kedly v/ithin 48 hours with concom- 
ittant profound reduction in secretion of cortisone and aldosterone 
equivalent to the depression produced by hypophysectomyo 

Dr, Ttillner is now studying a number of analogs of DDDo 
Difluorodinitrodiphenyldichloroetha.ne inhibits production of 17- 
hydroxycorticosteroidSo Diaminodiphenylmethane has an action 
similar to amphenone but produces no anaesthetic effects and has 
pressor acti-inty,, This indicates that the methylketone structure is 
not essential to an antiadxenal effects ^ut the coinpound is too toxic 
to warrent further invegtigation^ 

Transplantable rs-t adrenal carcinoma 494 is being studied by 
DrSo Ho Lo Stewai't and Ko Co SiielL Biochemical studies of this 
neoplasm by Dr., Do F,, Johnson (NIAMD) reveal an excess of steroid 
hormones similar to those found in the adrenal corteXo A substrain, 
494Hs derived by passage in hypophysectomised rats,, is morpho- 
logically distinct^ and its characteristic histology is maintained, on 
transplantation to intact ratSo It produces polyuria^, polydypsiaj 
degeneration of renal tubules, and causes hyperplasia of the ttiammar 
glands, which become filled with milky secretion in both sexeSe Th; 
uterus enlarges and the vaginal epithelium is mucifisd in female rat,> 
the testes and secondary sex organs of the male airophy^ Growth 
of this tumor in hypophysectonnized ratSj, however, induces develop-' 
ment of the secondary sex organs but not of the gonads.: 



rroid neoplasms in rats and mice represent a wide v^.. 

vuctural and biological type So Dr<, Ho Po Morris also finds t' 

iiiavior inconstant with respect to changes in znorpbology or g 

" ! e when stimulated by administration of thyroid hormone or p 

■ ';."f thyrotropin,, 



45„ 



Dto So Ho Woliman and Dr, Leonard Warren (NIAMD) found 
sialic acids 3, 9* -carbon amino sugar, in normal thyroid glandc A 
specimen of pure thyroglobulin provided through the courtesy of 
DjTo Harold Edelhoch (NIAID)s contained one percent sialic acid. 
Hence,, a broader study of sialic acid in relation to thyroid function 
has been initiatedg The highest concentration is found in the thy- 
roid glands of hypophysectomized rats, the loweat in glands xrom 
rats fed thiouracilj thyroids frorr) eutJayroid rats have concentrations 
intermedis-te between these two ejstremeso Administration of feMo-- 
uracil reduces sia.lic acid concentrations progressively j, with a 
minimum being reached, at about nine days„ This inay be due to the 
known resorption of thyroglobulin caused by thiouracilo Since some 
hold tlmt thyroglobulin iriust be changed by ensymes before thyroxin 
is liberated^ DrSo Wollnian and Warren have measured the concen- 
tration of free sialic acid which should be liberated by the same 
proteolytic process and find it to be highest at the time when con- 
centration of total sialic acid is declining most rapidly^ Free 
sialic acid content is lower in thyroids from hypophys,^ct.omized 
than from normal rats„ 

These basic observations have been extended to a study of 
experimiental thyroid tumors in rats by Dra. Woilman and Warren 
and Dxo So So Spicer of NIAMDo Colloid in sonie of the neoplasms 
is stained by the periodic acid-Schiff (PAS) reactiono Hornogenates 
of these tuinors are viscous, suggesting the presence of mucins„ 
Other thyroid tumors contain no PAS positiv^e mnaterial. Many of 
the tumors contain acidic mucins in which the acid group proves to 
be sialic acid rather than sulfate » Grovyth of one neoplasm in which 
sialic acid can be demonstrated intraceliularly is associated with 
increased leirels of sialic acid in blood serixm aiid urine o Some of 
the ixiore functionally active thyroid tumors contain neutral mucins^ 
as does the thyroid„ Autos-adiographic studies of a tumor with neu- 
tral mucins reveal that all follicles form organic iodine compounds j, 
whereas another functional tumor containing acid mucins in the 
colloid contained some follicles in which no radioiodine was detected,. 
The relation of acid mucins to thyroid function is being ciarified„ 



Hadiation biology 

From a precise and detailed study of the deleterious effect." 
visible light on a strain of haploid yeast ceils., Dro M„ Mo Elkind 
has turned to investigation of X-ray damage, and recovery in 



46c 



rnaminalian cells in culture o Initial studies have utilised two strains 
of cells from the Chinese hamsters Cricetulus griseus„ propagated 
in tissue cultures of the type originally deBigned by Pucko This 
permits the grov/th of individual cells as essentially clonal colonies 
whose morphological and biological properties can be connparedo 
Ejsperience to date includes detailed analysis o£ damage prodiiced 
by a single dose or fractionated doses of X-rays generated at 55KV„ 
and deliv^ered at the rate of 720 rads per minu£e„ 

These results are especially significant; 

l„ The vast majority of the surviving cells connpletely repair 
their accumulated damage before their first divisiors post-irradiationo 

Zo The kinetics of recovery depend on the physiological state 
of the cells and/or can be caused to appear to undergo large oscil- 
lations depending on the recovery mediuirio. These apparent oscil- 
lations niay result £ron:i coiinbined effects of changes in sensitivity 
and repair of inactivated siteSo 

3, Althoitgh there are important quantitative differences^ 
logphase cells respond similarlyo 

4„ A cell can undergo repeated cycles of damage and repair 
with no apparent attenuation of the repair procesSo 

As Dt„ Elkind and Miss Harriet Sutton state in their paper 
(Nature 184; 1293-1295, October 24, 1959): 

"There are several contexts in which these findings are of 
interesto If the chroix^ooomes are the X-ray sensitive sites and 
chronnosonrie breaks are the hits leading to lethality, then some new 
properties of restitution must be consideredg Firsts restitution 
goes to completion in surviving cellSo Secoiidlys the cell's ability 
to restitute breaks remains unijnpaired after repeated doseSo In 
view of the preceding, Puck's report of a high yield of mutant 
characteristics in the progeny of cells surviving five to seven m<b.s.n 
lethal doses may be applicable to the material he was using; rria-- bs 
evidence of a radiation -induced chromosomal lability which 
expressed after recovery and during clonal growth; may imply i.i,'c:.c. 
rnuta,t:ion pi'oduction and lethality are notj in general, closely con- 
nectedj or nriay indicate that the chromosomes are not the primary 
sensitive sites rsla.fced to viabilityo 



47, 



"Another area in v/Mch these results may apply is in 
connection with tumor therapy „ Treatmnent protocols iti'rolving 
fractionation are cominon„ permrfcting in general, ample tinne 
betv/een treatments for considerable if not coixiplet© recoveryo <, , o 
It recovery is not duly accounted for- the survival using xraction- 
ation can be higher than expected by several orders of magnitude » 
Of coursst tissue recovery in a general sense has been recognized 
by radiation therapists for a long time. These results, however, 
pro\!ide a. cellular basis for this phenoinenon a.nd lend specific 
direction to the ressas-ch that should be undertaken both to take 
advantage ofg as well as to controls this effects" 

Similar e:cperiments conducted by Dro Ro Z„ Lockhartu Jro ,, 
and Dr„ Elkind indicate that HeLa cells respond in a manner 
similar to those described for cultured cells of the Chinese hamster^ 
bat the HeLa line studied is not s.s stable,, 

Dro Ko Eo Bases has e^sploited the availability of single cell 
culture techniques to study the influence of actinomycin D on the 
response of HeJLa ceils to in vitro X-irradiationo He reports en- 
hanced lethal effects of X-rays on single cells exposed to the drug 
just before or after irradiation or diiring clone formationo Cells 
are more sensitive to X -irradiation when 85 percent of the water 
in the mediun^ is repla.ced by heavy waters and;, indeed,, the ceils 
can not survive indefinitely in the heavy water medium. Ceils sur- 
viving prolonged e^qsosure to heavy water ax'e abnormally sensitive 
to subsequent irradiationo 

A radioactive cobalt source is nov/ being installed to replace 
machine -generated X-rs.ys in the study of radiation chemistry by 
Dr, Co Ho Maswsllo Use ox gas chromatography permits isolation.. 
concentration and quantification of acetic acid resulting from 
irradiation of glycinco L-ike aminonia and foriTialdehyde, but -..^.^ !.-, 
glyoxylic acid fos-mation, acetic acid production depends on the 
concetitratioB of glycine in the radiated solution as do most organic 
soluteSo E:sarnination by mass spectrography of acetic acid formed 
by irradiation of glycine in heavy water suggests that the mechanism 
of acetic acid proposed by Weeks and Garrison is only partially 
correcto Only one-third of the acetic acid contains deuteriuirso On 
the other hand, Dro Maxwell has confirmed quantitatively the find- 
ing by Weeks an.d Garrison of substairtlal yields of aspartic and 
diaxr)inosuccinic acids „ One unknown compound re su^' ' 
irradiation of glycine remait^s to be identified. 



48o 



Information on the abstraction reactions of hydrogen atoms 

in the radioiysis of aqueous organic solutions is being obtained by 
Dxo Petez' Riesa, who joined Dr„ Maxvvell during the yeai'o When 
aqueous organic soiutio-as are e>:posed to ionizing radiation^ hydro- 
gen is pi'oduced by two distinct processeSo The primary molecular 
hydrogen yield arises from the recombination of H atoms in a series 
of small regions of high radical concentrs^-tion and is independent of 
added solute under certaiii conditions » Additional hydrogen is 
formed from the reactions of H atoms which diffuse out of the small 
regions and react v/ith the organic solute. Analogous processes 
occur when organic solutes dissolved in heavy water are irradiatsd;, 
and the primary molecular yield of deuterium is known from studies 
such aa those reported by Dainton„ 

Dro Eieas has studied abstraction reactions of certain amineSs 
ajiTiino acidSj amideSj peptides and thiols,, Deuterium atoms react 
with glycines acetamide and giycyl giycitie at neutral pH by abstract 
ting from carbon but not froin nitrogeno In strongly basic solutions^ 
an appreciable fraction of deuterium atoms abstract from the amino 
groups of glycine and n-butylarA-iineo The rate of hydrogen abstrac- 
tion for substances such as n-butylthiol a.nd cysteine is higher from 
sulfur than from carbon., In neutral solutions abstraction reactions 
account lor no more than one -half of the deuterium atoms produced 
by X-irradiation ., Sasults for glycyiglycine are particularly 
interesting in relation to radiation chemistry of proteins^ since the 
reactions of H atoms with the peptide bond will not produce free 
radicals with the odd electron on the nitrogen of the peptide bond^ 

Interest in effects of total body X-ii'radlation continues^ 
Dr„ Falconer Snaith reports that recovery of immune responses in 
xriice after two doses of X-ray given at varying intervals demon- 
strates relationships siiriiiar to those associated with lethalit^^ 
studieSo Multiple exposures axe cumulative in injuring the iriirrra'ae 
response even though no effect snay be noted on the leucocyte counto 
The recovery of transplantation immitnity of cells from, DBA strain 
mice in LAF]^ hybrids following sublethal irradiation and various 
treatments which accelerate heznatopoietic recovery is also urirela~ 
ted to recovery of the blood ceils <= Dro vV, W„ Smith, MrSo Joanne 
Koiicroft and Profo Jearome Cornfield (Johns Hopkins University) 
find that treatment with cells from imm.ature mouse sple:^~r ' - - ; - 
recovery; treatment with colchicine delays it. 



49o 



DXo Wo V/, Smith finds important differences in the protective 
effects against lethal total body irradiation of bacterial endotoxin 
on the one hand, and colchicine and its derivatives on the othero 
Endotoxins are effective in protecting youvig mice and still protect 
mice five v/eeks oldj, though hezriatopoietic recovery is delayedo 
They also protect irradiated guinea pigs„ The endotoxins produce 
their effects in guinea pigs only after the radiation has been giireno 
Colchicine and the derivatives studied so far by Dr„ Smith do not 
protect guinea pigs or very young mice but are acti've in mice five 
weeks oldo They act when given prior to total body irradiation but 
not during the post -treatment periods The protective effects of both 
endotoxins and colchicine are associated with more rapid recovery 
of the bone marrow among the treated subjectSo 

"Secondary disease" is a condition occurring in mice at 
inconstant, long intervals after exposure to lethal doses of X^irra^ 
diation and treatment with protecting doses of non-isologous bone 
marrowo The diseassg which is basically an immunological phenom 
enon, maybe fataL Miss Uphoff has pre\H,ously associater .iE.com» 
patibility at gene locus H-2 as a major factor in production of sacon- 
dary disease^ and now reports histoincompatibility at loci H-1 c.nd 
H~3 to be unimportant in the pathogenesis of the conditiono Other 
factors influence the long-term survival of the lethally irradiated 
and protected micso Miss Uphoff has investigated the efficacy of 
marrow from parent strain mice in the protection of F, hybrids 
produced in seven different combinationSo Early fetal marrow is 
effective in protecting irradiated hybrids in most of the combinations 
studiedo Marrow obtained from neonatal mice produces less severe 
secondary disease than is obtained with adult n-iarrowo When a gens 
difference is present at H-Zj only 50 percent of the recipients of 
fetal xnarrow survive secondary diseasco 



Natural hist ory of leukeznias 

Earlier annvial reports have included a nunfibsr of precise 
clinical and pathological studies on phenonnena associated with tli- 
leul;.emic states such as lesions of bone^ spontaneous bleeding an;" 
basis, intracranial lesions associated with hemorrhagSj intercui-:. 
infection and the immunologic response to specific antigens„ The? 
ha%'-e been achieved through the continuing studies of leulceniia 
patients by cliniciaiis and pathologists working together on commoi 
problems. 



5Gc 



DrSc Eo Jo Freireich and Eo Freij Ills ha¥e recognized a 
syndrome due to increased intracranial pressure in 25 of 150 
pallgnts, primarily children with acute lymphocytic leultemiao 
Eight o£ the 25 developed their first symptoms while in drug-indiaced 
remissionSo Studies by Dro L„ Bo Thoirjas and colleagues of the 
Pathologic Anatorr^y Branch associate the aymptoms with an internal 
communicating hydrocephalus and extensive infiltration of the pia- 
arachnoid by leukemic cells which in extreme cases obliterates the 
subarachnoid space over the spinal cord and braiuo This is not a ' 
completely new lesion^ but review of the literature reveals a sharp 
increase in the occurrence of the syndrome since the advent of 
chemotherapy o 

It emphasizes the need for more intensive investigation of the 
blood-brain barrier^ as approached by Drs= D„ Po Rail and C G, 
Zubrodo This physiological or biochemical barrier develops early 
in life since it is already present in puppies 5 to 20 hours old„ 
Neither hypotension nor administration of cortisone increases the 
entry of drugs into the cerebrospinal fluido Sulfanilic acid^ a strong 
acids fails to enter the cerebrospinal fluid in the dogfish at 96 hours 
in inore than traceSo Para-aminohippuric acid^ a weak electrolyte 
of which only one part in 6000 is dissociated at body pH^ will achieve 
equilibriuni in tirna by diffusiono Coixiparison of the ability of these 
two compounds to pass the blood -brain barrier further radicates 
tliat undissociated ra.ther than dissociated dr-ags will penetrate into 
the cerebrospinal fluid and si;rengl:hens the lipoidal concept of the 
barrier. 

Comparison of the entry of antipyrinej, sulfadiasine and 
para-aminohippurate into the cerebrospinal fluid of .man and dog by 
DrSo Eall and Zubrod shows the same basic phenomena in both specie 
The humanj howeveri, approaches equilibrium at a slower rate^ 

While methotrexate does not readily enter the cerebrospinal 
fluids i5r„ To L.O Loo has detected 6- mercaptopurine in cerebro- 
spinal fluid of dogs receiving intravenous injections of the drvi.g„ 
The spectrophotometric identification of the compound has been 
confirmed by paper chromatographyo 

Dr„ George Brecher (NIAMD) and Dr„ L, E„ Schroeder,, Jr., 
are studying synthesis of deoxyribonucleic acids in hunnan leukemic 
cells by an in vitro techniques utilizing tritiated thymidine and 
radioe.utography in such a v/ay that no new cells enter the synthe'-ic 



. ■■j&e q1' the gisU*st,'&tion cycle aad none divides, durwig ih"- 

,:■ : .{y". rhey observe no coyrelat-aoR between deowfrihon j. 

' vr.thetic activity a?id the usual morphological crltfss'is 

in r, am?ityo Alcfcough & v^ide range of rates o£ synthesis. - =. ■;.,. 

they do -not eKceed the zang© exhibited by ietacocyfees Iron'. iTaittenr 



■ iibucK ;;-k^n = \. iadovv technique is, being etriployei'i V: 
a stixdy 6/ e,xper-'.nientaJ. inflariinnatioti .m patierjiK^ 
acute ieviKgmiao Frequenily tixes® patients ial* to rnobilize p 
phoojiClf-Ar )ei,^c. Of A' ■/?■■■" J?i ,">- y-i o ■>•..:- ;o injury « This abnormaJ 
r=^;-;: .T ' ased susceptibility to inft 

and tn svjj:T-ti degi'r; , ;- ■r^-^i.-Avo lo vne numbes' of polymorijho-miic strg 
leucocvtfis in the cii'cula.iine bloods 



hypoganvrnagiobuj >nsrrc?a i.r.! patiems with c.b.rom.c iymphocyttc 

;<;>r;:i- !"•-<:;'-. fl .f,,r- :. ^. - " :, - :' r oiT) pauied by dcD '"-^^ < ' ^^''•■'"' --^f '''«" ^-r 

■.iscific antigenic 

uA aatia.oftV p70<;:^.;c^^::J:•J S'Sems to relate mort c; i r^c f f: dd.^;' 

susceptibility of infection than to t&e concen- v alobisl'm* 

the serum,. Dr, J. Po Uv.."?. (NIAID) has colk; 

in fhe •ir>-rr-iir;o]o'?^r,~] '-^"r^lv-'S, 



i :-'-:' idomonas sepsis and clostridial Infe. 

coMinue ■o ::>-• i^o..-. ' ::;rospective study of moniUaf 

ff-v;?eals thrush in i . 148 patients „ Almost ?>M of .' 

T'-.V^ r'.ve coninienc&ci during the last two moi-iths o' 
i .l: , ■ ihe episode has been significantly dirninished 
r oxx^Ja washo While antibiotic a.nd corticostei'oic 

■ ojina t»atier'' 



a piasrr 



>2, 



■Anemia and gelated dis orders 

A patient with cerebellar hema,ngioblast.oma and polycythemia 
admitted during the year was studied by Dto T, Ao Waldmanni, who 
described marked stiiTiulation of erytkropoiesia in the rat by a non- 
dialyzabie. factor obtained from the contents of tha cystic portion of 
the neoplasiric This appears to be the fii*3t demonstration of eryth- 
ropoietin -like activity in any biologic material other than ux°ine or 
plasma and supports the concept that poiycytherniap not infrequently 
associated with cerebellar hemangioblastorrias, xnay be ca.used by 
production of an ei*ythropoietin-like factor by the neopiasnio 

Drp Bo Eo Greenfields, Jr, , has ejctended his studies of 
erythrocyte fragility in relation to age of the red blood cell popuia- 
tion„ Erythrocytes more than 12 days old hemolyae at 0„ 4 percent 
sodiunn chloride while younger- forroa v>fMch hemolyse at a concen = 
tration of 0,2 percent can be further separated by ejcposure to gradl« 
ents of salt content approaching 0,3 percsnto The absolute values 
vary soinewhat among specie s„ Systematic study of red blood cell 
repletion in rats rendered a.nerjnic by repeated bleeding reveals fra- 
gility values for immature red blood ceils and reticulocytes of Oo i 
percent^ three -day-old erythrocytes of 0.2 percent while hemolysis 
occurs at Oo 3~0o4 percent concentration when the red blood cells 
are 12 days oldo 

Dr„ Greenfield and Dr. Vo Eo Price have also shown the slow 
disappearance from the site of deposition of erythrocytes labeled 
with radioactive iron when injected into a transplantable tumor or 
into subcutaneous tissue^ The results are coimparable to those 
reported last year for studies of labeled erythrocytes injected into 
skeletal muscle „ 

Having previously described the loss of catalaae activity by 
parenteral injection of 3-amino-l, ?-, 4~triasolct whichj never'thelesSj 
does not interfere with synthesis of the ensymej, DrSc V, E, Price 
and Miloslav Eechcigl (Besearch Fellow) report kinetic studieSo 
Although the rate of catalase destruction by am.inotriaaole is the 
same in both liver and kidneys the rate of ensyme synthesis ia four- 
fold greater in the livero Eventually an equilibrium is attained at 
v/hich destruction fcalsnces replacen-enf,, 

Dr, Waldii-.ri.tiii trf a.li,ei;;ipi.itig (O pi'oduce hyperspleiiiii-ii: ia a-,.;;V;^ 
by Intravenous i-.-ifusion of methycelluloseo Moderate splenomega.Ur 



is obtained togetfcer with an anemia in which life span of the 
erythrocytes is reduced,, The picture is complicated by severe 
renal damage and ui-emiao Administration of large doses of desic-' 
cated thyi'oid increases taie dog's blood volume as much as 2 5 per~ 
cento Increased synthesis of erythrocytes can be demonstratedj 
but their life apan remains within the normal range. 



it l0£ 



An increased requireiirsent for sodium ioa.s was reported last 
year by Dxo J<, White as characteristic of rats bearing progressively- 
growing Walker carciiiosarcoiTia 256o He, with Dr^ Fo K„ Millar 
and MrSo Jo No Toaij has studied the metabolism of sodium by nor- 
mal and cancerous rate by balance techniques. The normal subject 
excretes sodium in relation to the amount protrlded in the dietp but 
the cancerous rat reduces its sodium excretion 15 to 20 days after 
the Walker tuirjor has been implanted even though the animal is stor^ 
ing nitrogetio Chloride and water follow the same pattern as sodiuiT5» 
but potassiu:m irietabolism is not affectedo When the diet contains 
adequate announts of salt, and this a considerable excess over the 
requirement of the norjnal rat, the cancerous subjects lose little 
weight despite their large tumors and the adrenal glands do not en- 
large. The cachectic tumor -bearing rats whose sodium intake is 
restricted have marked enlargement of the adrenals affecting parti- 
cularly the aona glosTierulosa, In i-ats eating a diet va which lyophil- 
ized tumor forn-^s the only source of protein^ the excretion of ailan- 
toin is inversely related to the excretion of sodium^ though such a 
diet is rich in salt and nucleic acids, 

Dr, White and colleagues have also studied sodium excretion 
of rats bearing the Murphy -Sturm ly/nphoma. The diet v^ith restric- 
ted salt coiitent results in initial fall in sodium excretion as the 
neoplasm growSj but the level then rises and is maintaiiied as in 
normal rats,, and the adrenals do not enlarge, Sozne tumors niay 
have so great an 8Jd;racelluiar space tha,t the ordinary diet Siiffi- 
cient for growth of the normal rat cannot fulfill the needs of a 
greatly ejcpanded extracellular space. It seexns quite likely that a 
need for additional building blocks to manufacture proteins or 
nucleic acids for a. rapidly growing protoplasmic ma.ss might have 
equally deleterious consequences on the host. This general subject 
will foe essolored further. 



54„ 



At any rate^ the characteristic growtb, pattern of the Walker 
tumor and similar consequences have been described in modified 
form for rat lymphosarcoma R2788 and rat hepatoma 3683 by 
DrSo Hechcigl and Greenfield, 

Dr„ Pratt finds it possible to determine the gross body 
composition of cancerous rats in viyo through serial calculations 
of ratios of nitrogen stored or lost to caloric e:£penditure„ Pre- 
liminary data indicate severe gross changes as would be anticipatedo 
The direct calorimeter for the rat„ designed by Mro W^ Co Whites 
has required additional modifications but should be in use in the 
near future. 

Measurement of changes in body composition seems a 
reasonable approach to the study of cachexia in cancerous patients 
and may lead to some ineans of describing the changing mass of a 
tumor which cannot be seen or n^easured by more conventional 
means. Additional experience in the measuremient of changes in 
body fat in the Siri apparatus by Drs, Berlin and Watkin^ correlated 
with metabolic balance techniques and independent measurennent of 
body v>;aterj, reveals excellent agreement among the values obtained 
by the different methods, 

Dr, Watkin has characterized a "v/eight loss syndromcj, " 
which includes negative caloric balancej lov/ respiratory quotient 
and increase in unesterified plasma fatty acids„ which he asso- 
ciates with actively growing cancers. Study of the effect of fasting 
reveals that the patient v/itli rapidly advancing malignancy either 
exhibits this syndrome at the beginning of the experinnent or changes 
to that general pattern within 12 hours. Such persons whose basal 
metabolic rat6 is high initiallyj contiirae to manifest the same high 
rate despite fasting. Normal individuals, on the other hand^, show 
little evidence of a "weight loss syndrome" until 16 to 36 hours 
after the fast begins. 



Tissue culture 

Measiirement of several important biochenaical qualities by 
Dr„ B, B, Westfall points again to the high degree of chemical 
variation among cells grown in vitro for eidiended periods. All 
of the ce'iT ^-'-'^^-vi a'.-iTni^.-] form a'-keto s-cids. Concentrations of 



55, 



nucleic acids, glycogen, lipids and cholesterol vary widely from 
sti-ain to strain. No glycogen can be demonstrated in cultured 
fibroblasts; yet after 7 years in vitro a line of skin epithelium 
stores glycogens, as do tv^o strains of hepatic parenchymal cells: 
cells of a hepatoma strain derived froin one of these hepatic paren- 
chymal strains, on the other hand, have lost that capacityo In 
certain cell strains ensyme activities have been lost; in one cell 
strain a very great increase in arginase activity was demonstrated„ 
Certainly cells in ra.pid proliferation over long term in tissue 
culture show numerous divergent variations, 

A strain of monkey kidney cells obtained originally fron^ Eii 
Lilly Hesearch Laboratories has been adapted to grow in a chemi- 
cally defined mediunn by Dr, V, J, Evans, This strain supports 
growth of poliomyelitis virus very well, Dr„ Evans has adapted a 
contiiiuously cultured line of human skin epithelium to growth in 
chemically defined medium by slowly reducing the concentration of 
serujn in the original mixture, A nev/ medium NCTC 117, derived 
from the familiar NCTC 109, omits coenzymes, sodium giucuron- 
nate, deosyguanosine, deosyadenosine, 5 methylcytosine and 
"essential" unsaturated fatty acids. The only nucleic acid deriva- 
tives required by strain L are thymidine and deoxycytidine, 

Dr, Sanford, studying the effects of vitamin requirerricnts of 
cells growing in the chenaically defined medium NCTC 117, reports 
pantothenate, choline chloride, niacinainide, thiamin and r'iboflavln 
essential for cell survival. Folic acid, pyridoxal or pyridoxine, and 
possibly biotin, while not essential for cell survival, increase the 
rate of cell proliferation. 

The effect of methylcellulose in iiXiproving cell growth in 
chemically defined n-jedium in agitated fluid suspension cultures is 
not related to its viscosity, Mr, J, C„ Bryant, Dr, Evans, Mr^ E, J 
Schilling and Dr, W, B, Earle find no notable growth promoting 
effect, in such cultureSj of widely different concentrations of 
methylcellulose on ceils of a monkey kidney strain. 

Study of the characteristics of cell populations growing in vitro 
is made possible by coxnpa.rative time lapse cinematography. The 
first study hy Mr, W, T, McQuilkin and Dr. Earle concerns changes 
dviring adaptation of one clone of strain L to growth in a protein f r f u 
medium. The avera.ge generation time is prolonged and the migra,^ 



56. 



rate on the glass surface is greatly deca°eased(, as is the cytokinetic 
pliase of growtbo The cells are less compact, refractile and gran- 
ular, Thsy round up only prophasco Monkey kidney cells studied 
under the same conditions demonstrate a generation time similar to 
that of fibroblasts, and the population increases 24-fold in a weeko 

Lymphoma P 388 groxvs well on Eagle "^s medium plus 5 percent 
calf seruirio Dialysis of the serum destroys its capacity to support 
growth of these tumor cells when added to Eaglet's nnislureo 
Dro Robert Roosa (Research Fellow) finds that addition of pyru-rate 
or L-serine to the diaiysed serum restores its growth promoting 
properties., 



Cytochemistry and histochemistry 

A number of cheinicais cause blabbing of cells under in vitro 
conditionSo Dr„ Mo Ko Belkin has extended his study of blebbing 
to the extent that he considers this to be a common property of n.eo = 
plastic cellSo Experience with cells from 5 normal tissues fails 
to reveal bleb forrriation by the same agents that produce the change 
so readily in cancer cells. The majority of compounds producing 
blebs form irjercaptide iiiikageSf but a few oxidizing and alkylativig 
agents also display the saixie propertyo 

DTo Robert Love can recognize 9 morphologically distinct 
formis of ribonucleoprotein in cells stained by his fcoluidine blue 
molybdate techniqueo Electron dense molybdate is deposited at 
sites of metachromasiao Some hepatic ribonucleoproteins are 
affected by starvation; some disappear altogether and reappear on 
feeding, Ribonucleass in concentrations of 40 mg per mU has no 
effect on the staining capacities of cells, 

Dr, Love can demionstrate a nucleolinus in a number of 
normal and neoplastic cells „ Although it is usually large in cancer = 
ous tissues, a -rariety of other unrelated conditions may also cause 
it to enlarge. Availability of the toluidine blue molybdate technique 
has pernmitted a study by Dr„ Love of the effect of colchicine on 
ribonucleic acids in cells, Metaphase arrest, characteristic of 
colchicine action, is associated with failure of nuclear parachrom - 
atin to diffuse into the spindle zone at the end of prophase, Soitie 
di'^aes of the drug resuli: >n corr.pleti? inhibition of mitosis with incr?; 



of parachrojpDatm durir,; .,:base and • the ttu,eleu® 

and nacieolfeaSo Paracbi^ornatin may play soir.- the ririgit. 

of spindle fibers,, 

The varciation .: eaction oi aaciies tumor ceiis ia tetra = 

2;olium salts varlef; :lj that Dr, G, Zo Wiiliarr.s h».& turned 

Ms attention to study c hepatic cells from mouse and rat, *hich be 
has learned to separate aed to count electronical lyo Isolated mito- 
chondria possess a variety of dehydrogenases demonstrable by 
tetrazolium techniques, biri. mitochondria In whole cells react quite 
differentiys as DTo MacCaifdle has been ©aylng lor years,.- Che 
differences in reaction rates observed in vyhole cells, as corppared 
with isolated mitochondria, are probably not due to differences In 
permeability, because addition of a rapid reducing agent after 15 to 
30 minutes of eseposure of intact cells to otig of se-vferal tetraisolium 
salts causes rapid reduction of the intr?,' 

Df:, Williams has calculated redv: -or severai 

tetraaoiium-siubstrate combinations „ Concern rations of 0,., OOli^ M 
tetrazoiium and 0= 034 M succinate result in rapid intejnse reductior* 
to formasan in cytoplast-n of hepatic parenchymal cells? near the 
mitochondria with gens: .:,I increas© In cytoplasmic densityo l-ower 
concentrations lead mulation of formazan in lipid droplets c 



iJern^atology 

Dr.. £0 Jo Van Scott reports that autotranspiantation of ep 
tissue free of stroma from the donor sit© is either unsuccessful oj 
the transplanted cells grow .nd assume the characteristics o:-' 'he 
epitheliuni of the new site.. The cells will not grow in the su 
Cells from a basal cell carcinoma will grow in a nev '•■'^ --^ - 
donor stroma is available in the transplanted fragm. 

His continuing studies of psoriasis include . 
nursiber of agents which cause pronnpt clearing of the local i- 
topical applications with recurrence in one to severs T v/f .?!- ■ 
drugs as rjriethotrexates 5-£luorouracil„ actinomycir. 
all active against some neoplasms, will clear up the . - r 
when administered systemicallyo Antimetabolites are iiiacl 
applied locally „ Serial histological studies of the le- 



Nine protein fractions „ three fractions each isolated by 
Identical methods from human epidermis^ psoriasis scale and 

ichthyosis 3cale„ hav^e been shown by Ds„ Simon Rothberg to display 
major similarities when their peptide patterns obtained by enzymatic 
hydrolysis art; compared, Soltibiiisation o£ the proteins does not 
require cleavage of disulfide bonds^ Some peptide differences liave 
been observed am.ong proteins obtained from normal epidermis„ 
psoriasis s, and ichthyosis„ 

Additional work on the relation of hair growth to the dersnal 
papilla is reported by Dr„ Van Scott and Ds^ R„ G, Crounse (Research 
Fellow}„ The mitotic activity of the germinal rriatrix is proportional 
to the number of cells in the papilla^ normally 1 mitosis per 8 papilla 
cellSo Mitotic activity decreases in successively higher levels of the 
hair bulb but is a function of the area of the papilla to v,/hich the matri:r- 
cells are exposed and the transverse thickness of the matrix at each 
levelo Permanent baldness results from destruction of the papilla, 
Dro Crounse and Dr, Sotliberg find arginase in the hair sheaths but 
not in the bulbar portion of epilated roots o This suggests that argin- 
ase is not directly concerned with synthesis of keratin of the hair shaft„ 
While arginase is known to exist in the epideriTsis, other snaymes of 
the KrebstHeaseleit urea cycle cannot be demonstrated in this tissue^ 



Detailed study by Dre, Mo K„ Barrett and E<, Jo Breysre 
(Research Fellow) of transplaiitability of l;umors among several com- 
binations of instrain and outcrcss matings provides sonne unexpected 
resultSo Among females of a given or first strain of micej those 
impregnated by a male of a different or second sti-ainj exhibit toler- 
ance towards a tumor originating in the second straino This is 
exhibited by successful transplantations of the neoplasm in immu- 
nized fennales mated to males of the 3tra.in from .which the tumor 
originated, but not in those mated to other n-saleSo The effect inc;; 
with increasing multiparityo 

Dro O^Gara is transplanting thymic tissues of newborn rxiice to 
the spleens of adult miceo The thyj'nus regenerates in this locationo 
Adrenalectomy and orchiectomy enhance the growth of the transplants,, 

but ovctriectomy a.nd thymectcniy produce no effect. 



59, 



Some cancers clear glticose from the blood quite rapidlyo 
Dr, Ho A, Kahlei% studying the effects of glucose administration 
on pH of an expesfimental neoplasms has used both intraperitoneal 
and intravenous routes of administration. The former causes flow 
of fluid from the blood into the peritoneal cavity with dehydration 
of the tissueSj, while the latter increases the blood voltHue includ- 
ing blood flow through the tusirsor; hence produces a ztiore rapid 
drop in pH of the neoplasm and a more rapid return to normal val- 
ue So The reduction in pH following intravenous injection of glucose 
is Oo 65 pH in the viable part of the neoplasm, but only Oo 16 pH in 
its necrotic centero 

Dto Eo Do McLaughlin has standardized study of the rat liver 
regenerating after partial hepatectomy so that reproducibility is 
adequate to permit quantification of some aspects of the process., 
Preliminary results reported by Dr„ McLaughlin suggest that nor- 
mal hujnan serunri contains something that retards orderly regen- 
eration, whereas sera from cancer patients has no such effecto 



SURGICAL AND VIRUS TREATMENT OF CANCER 



Independent naeasurements of circulating plasma and red 
blood cell volume by modern isotopic techniques have been applied 
to study of the post operative state by Dr, A„ S. Ketcham and other 
members o£ the Surgery Brancho Once more the unreliability o£ the 
hemoglobin, hennatocrit and erythrocyte enun-seration as determined 
by conventional methods j to reflect changes in the effective circula- 
ting blood volume is emphasiaedo During the first two weeks after 
a major surgical opei'ation, a patient may lose 25 percent of his 
volume of circulating erythrocytes without any remarkable change 
in hemoglobin concentration or in hematocrito Tachycardia occurs 
when the effective circulating volume of red blood ceils is reduced 
by 30 to 40 percent, and these patients respond dramatically to 
transfusion of two units of bloodo 



60„ 



Review of surgical experience .at the National Cancer Institute 
includes 245 major operations and 214 minor surgical procedures^ 
Only 7 patients have died within 30 days of operation^ There have 
been 44 post operative infections of which 20 have been associated 
with avascular operative wounds caused either by extensive X-irra- 
diation of the site at varying periods before surgery or by the 
creation of lai-ge skin flaps as part of the operative procedure » 
Antibiotic -re sistants coagulase -positive hemolytic staphylococci 
have been major factors in 29 of the 44 cases, but an additional 17 
patients known to harbor such organisms have not developed wound 
infections post operativelyo Nineteen of the 44 patients had contam- 
inated wounds that involved lesions of the mouthj pharynXj trachea 
or the gastrointestinal tract before any operation was performed,, 
This experience led Dr. R„ R, Smith some time ago to design a pro- 
spective study of post operative infections and it is already clear 
that 23 of 30 patients harbored at the time of operation the organ- 
isms which subsequently caused post operative infection. 

Continuing study by DrSo Smith, J, F, Potter and Malmgren 
reveals that the contamination of wounds by cancer cells occurs 
most often after operations that involve excision of a primary cancer„ 
Neck dissection for metastatic cancer alone seldom results in the 
recovery of recognisable cancer cells from wound washings^ The 
frequency of contaminated wounds in operations for epidermoid car- 
cinoma continues at 26 percent and no correlation between contam- 
ination and local recurrence is possible. Use of dilute solutions of 
formaldehyde has been ineffective in reducing the frequency of local 
recurrences. No new leads have developed in the experimental 
study conducted by Dr, Ketcham in a search for agents which may 
kill the residual cells. Of the several chemicals tried, all promote 
the growth of cells when the wound is treated before introduction of 
the brie, and thiotepa has a similar though less pronounced effect 
when given by intraperitoneal injection. Any one of the agents studied 
will kill cells of experii:nental tumors when suspended in the sar/ie 
concentration used to wash the wound. The wound is simply a different 
and more complex environment. 

Cancers of the head and neck are commonly said to metastasise 
to the organs and tissues below the clavicles in only 12 or 13 percent 
of patients. Quite a different picture is afforded froixs limited exper- 
ience with this type of inalignant disease at the National Cn^icer Insti- 
tute, Dr, Snnith has reviewed the 25 post mortem examirations 



6l< 



perforBned by the Pathologic Anatomy Branch oti patients who have 
succumbed to cancers originating in 12 different sites of the head 
and necko Almost all of the lesions were epidfirmoid carcinomas, 
and one -half of thein had not metastasised at t'ne tinne of the opera- 
tiono Thirteen, a little more than half, of these 25 patients had 
distant metastases involving 17 different locations^ and in 12 cases 
the lungs were involvedo Three of the patients with disseminated 
disease never had metastases in the cervical lymph nodes. The 
average interval hQtweerii operation and death in this small group 
is only 10 months for those patients who had no clinically demon- 
strable evidence of metastases at the tizxie of operation and 19 months 
for those whose cancel's had spread beyond the site of origin when 
surgery was performed. Each of the lattex* series of patients v/as 
treated at least once for local recurrence. The prolongation of life 
among these people can hardly be attributed to the surgeon's skillj 
since they would seem to represent a somewhat more indolent type 
of cancer than characterized the patients whose life ejcpectancy was 
materially shorter.. 

Urinax'y diversion via an ileal conduit in radical pelvic 
exenteration for cancer of the uterus continues to give satisfactory 
results. Infection of the upper urinary tract and hyperchloremic 
acidosis are distinctly unusual in this group of patients, as con- 
ti-asted with their frequency in patients with wet coiostornies„ 
Dr<, Smith and colleagues observe a ureteral reflux when radio- 
opaque materia.1 is introduced into the ileal pouch, and bacteria.! 
contamination of the ureters is relatively commono 



Virus treatment 

The use of a strain of Coxsackie virus B-., trained by Suakindp 
et al, , to destroy HeLa cell tumors growing in rats, in the treatment 
of advanced huinan epidermoid carcinomas of the cervi:E (Annual 
Report, 1957), has been studied for several year5» The enterprise 
has in%rolved niany people, including Drs. E, Ho Smith, S„ B. Couch, 
Manaker and Love, NCI, and Drs„ B. Jo Huebner and W» P„ Rowe 
{NIA.ID)„ Although this sitioldering effort is not likely to break into 
flame at this time, experience v/itb. 25 patients is interestingo 

lo Among nine patients whose serum contained no antibodies 
against Coxsackie Bo tio difficulty was experie-aced in recovering 
virus during the first four days after its injection par vaginam into 



the ca-'iicexouts r. 
through 8 as die 
from only 3 pa.tieai.c. a. 
eleventh, day in only or 
value calculated to- 
oricmallv- Amn-n::. 



: ppea precipxsouaiy on days 4 
r viruSo Virus was recovered 
..as eighth day an4 persisted beyond the 
'3f;, D?Fp1te thir* experiences the total 
-eeds the amount injected 
- c^-i-r.-mad anfeibodie? 
impossible 



positi'^ 



for t?^' 



ana anus vv: 



in all patients by tli-:' 
b-?;:ween 256 and ' "' ' 



ers rose to 



3„ Fr - th 

2 5 patients o ^?d^ 

hours after injectio-a 
^ive lymphoc"''^'--"^ 



rtii'actiOii obsci\ed in 12 of tb'^ 

o5 toicm ar.cl occerred 12 to 



Injsci;icn o.. oo^.^ 
oX 4 patients produced 
hours.; which slo'i,vl"- 

ne'sati'ie for 1;^. ctsv% 



-.':..SHO 3,Bd C 



irc.ckie viruSv=s into the cancer r. 



■-'•vieTaA rrisl?>?3e arid fe^^^er of 39-40 C in I,? 

od cultu- 






ologic or histologic 



passage in vivo „ The o^'iginal chick eirjbryo strain produced 
some oncolyais of lymphoma P388 ascites tumor, but the Ehrlich 
adapted strain had no such effecto Dr. -Love concludes that adap- 
tation of a virus to a given tumor does not confer oncolytic power 
against other neoplasms. 



Pyridoxine deficiency 

Reports by others indicate that pyridoxine deficiency interferes 

with immune responses, and Drs, Couch and Smith conceived the 

idea of enhancing the oncolytic effect of adeno or Coxsackie E 

3 
viruses on epidermoid carcinoma of the cervix by clinically induced 

pyridoxine deficiency. This was accomplished by formulation of a 
semi -synthetic diet vvfhich included the anti -metabolite desoxypyri- 
doxineo Reports of clinical pyridoxine deficiency induced at other 
research centers guided the loca.1 study and, in general, each of the 
4 patients studied at the National Cancer Institute pursued courses 
consistent v/ith the S3m.drome described by others. The mental 
symptoms were different in that mood sv/ings with periods of stabil- 
ity and orientation were more frequent than expected. Only two 
patients exhibited marked lymphopenia. Marked impairirjent of 
z'enal function, elevated fasting blood sugar and profound catabolic 
responses seen in each of the patients are new findings. Perhaps 
the hyperglycemia may be associated with the known toxicity of 
xanthurenic acid for the p cells of the islets of Langerhans, 

The e:;cperience is too small to provide definitive information 
on enhancement of oncolysis by Coxsackie B-, virus by pyridoxine 
deficiency. Although signs and symptonns 02 the clinical deficiency 
v/ere reversed promptly by replacement of pyridoxine for the desoxy- 
pyridoxine in the diet, more information on some of the previously 
undescribed effects of pyridoxine deficiency should be sought from 
animal experimentation before further clinical studies are undertaken, 

Drs., Dyer and Morris, having extensive experience with 
pyridoxine deficiency in rats, have studied biochemical aspects of 
the clinical syndrome. The urinary excretion of several tz'yptophan 
metabolites is almost identical in man and rat during pyridoxine 
deficiency. The urine contains large quantities of xanthurenic and 
kynurenic acids, as well as large amounts of 3-hydroxykynurenin 
and its acetyl-glucuronide and ortho-suifate derivatives. These com- 
pounds, absent from the urine before induction of pyridoxine defi- 
ciency, disappear when dietary desoxypyridoxine is replaced by 

vitamin B / „ 
b 






''J.e>r! ■=<■*-"! 



65:,. 



DrSo Jo R= Anaj^..-. „ ;, „ L, Swarxn and a. i..t. ...,.:»,.., i. ;>*.. o 
studied another patient with extensive chondrosax coma o£ the 
thoracic wall whose neoplasm concentrated radioactive sulfur, 
A therapeutic doae of 388 mc of radioactive sulfa I e was followed 
after t,wo months by another dose of 335 inc, Te;mporajfy :retar = 
dation of tumor growth was followed later by inci'ease in growth, 
requiring eseternal beam irradiation therapyo Thi? isotope treaf- 
rr.ent produced on:" ?■■-■" aht depression of leucocytes and platelets 
until 4 veeks r_ . mistration of the second doscj when throni 

bocyfeopenia becanvs -narked and leukopenia moderatec The blood 
picture returned to nori-nal in two montha, but f> months after the 
second dose of isotope the patient developed a severe anemia and 
thrombocytopeniao Another patient treated in much the san^e way 
3-nore than a year ago has developed no such syndrome which re- 
calls e^rperienc'^ t-.o-o,- -? .^vri •;--. .-\,-^r,^. ?-,r n-.. Tsyor.he--^ some yea^s ago. 

Dr., D„ Pa rsc;..\;.dy ar:? sruftvec. nnetaboiic eitecti? of 
X -irradiation in a patient with a lymphosarcoma of the lower 
extreiifiityo Prior to treatment^ the metabolic pool o.f nitrogen 
was slightly increased, but no change in the rate constant of 
metabolic pool turnover could be detected, A dose of X-rays 
amounting to 1000 r was followed by marked enlargement of the 
pool with decrease in the sairie constant. Incorporation into body 
protein of amino acids labeled with N , ho unchanged 

during this period. 

In the ex r aborator; uit has studied the 

influence of increased oxygen tension on the response of trans - 
plantable adenocarcinoma in C3H/BA to local X -irradiation. He 
reports no enhancement of therapeutic effect of the X-ray treat- 
ment when mice bearing this neoplasm, ai-s exposed to an environ - 
irient of pure oxygen at two atxnospheres during X-irradiation, 
as compared to an environment of air at one atmosphere „ The 
result is the same with tumors measuring either one or two c.=^:.: ' 
meters in diameter^ A statistically significant differer;/. 



;^he i'£3ig^ial-.oi;;;j J.u.riiig ii3.'2 year oi Ivir^^ Ho •!<..:. oil and 
.at require complete rebuilding of the laborato?-" cempo^ 
programs relating to therapeutic use of ionising 

\<icri]fv in r.= eru?ti:: ■? Puils^bl- oiiiexii.s for clinjca; 



CHEMOTHERAPY OF CANCERS 



Critical analysis of experience obtained throughout the world 
in the drug treatment of cancers reveals a small group of specific 
anatomical types of neoplastic diseases in which chemotherapeutic 
agents have produced definite beneficial effects ^ and a inuch larger 
group of cancers which remain relatively unaffected by systemic 
administration by chemotherapeutic agents. There are, therefore, 
at least two separable, broad, general problems with a.n infinite 
variety of specific subordinate problemSo The substantial progress 
made in treatment of leukemias, lymphomas, choriocarcinoma and 
adrenal cortical neoplasms emphasizes the desirability of quantita- 
tive coiTiparlsons of the efficacy of chemotherapeutic agents already 
available, fui'ther exploration of chemical relatives of the effective 
agents for greatly enhanced therapeutic acti-\dty and research 
designed to elucid3.te the snost effective means of administering 
the drugs now on haiAd, A search for new drugs with qualitatively 
different types of activity marf also benefit pa.tients with responsive 
types of cancerj but this becomes of paramount importance to 
patients siiffering from the much larger number of neoplastic 
diseases which respond fleetingly or not ax ail to the currently 
available drugs. A variety of transplantable anims-l tumors is 
available for this re sear ch^ and probably the x-ariety nnust be 
increased if the necessarily empirical approach is to be fruitful. 
Increased emphasis on biochexxiical investigations with a view to 
categorising individual or special types of cancers seerms practical 
and desirableo Certainly a much greater use of patient nnateriai 
will be necessary before meaningful correlations between the thera- 
peutic reaction of man and experimental animals can be established 
andj indeed; we probably do not know how to study therapeutic 
responses of cancers of the stomach, large intestine or kidney at 
this tinne %vith any degree of precision. While the history of thera- 
peutic research is sprinkled with episodes as drannatic as the peni- 
cillin storys most of the ma.jor advances have required slow exploi- 
tation of small ga.ins by systematic study. 

The course of any research requiring creation of nev/ knowledge 
is largely unpredictable. Having pointed out the possibilities of 
exploiting certain iinportant developments obtained from the study 
of plasmacytomas, it seems appropriate to recount progress in 
correlated clinical and laboratory research in e. different kind of cancer 



■ '.-'Li froji' ba;sic injro'rmation oi' 
atfXi.Sie and fetal nietabollsn-i, and, inckiBda Lue xujtlaUy 
ifcerap-eutic regimen d.e.3i,i^n^-^d b" Dr., Tlertx aiua s>f M,. 
o.'viated stgnuicaT. ■ 



prkj"r to Septen-ber 15; .rA>::&l„ radi-^Hi-yV;; ■ 

o.e hormonal evidence ai i^5;&iduai disease iV.-i' p^gyiods rangir ■ 
frua^ 5 to 47 n'.uaths<, Eight have p?r'"~".~?-efv; clioriocarcirvin - 
' > have died iollowing an initls ■ ? hotrexate Efeerapy 

'Sth subsequent deve- 1 '-•■-"■ "■-■■■■ -^ -distance;.-, Acquirf 

drug resistance is . ■ e effectlw t:!,>a,na,g.: 

n-^ent of iMs. neoplasm, Metnoi ^ r:;ia..e vesjMi.^.nv patients d... vtofe 
respond to gabsequent trea.tn.Bnt with nitrogen njuS'tay d„ <vyfei>«®.?i 
Do 

:>asr.e May i959s i^2'« Herfiaanda: 

V' 1 vi c .; J - V :-: .-biastine , an alkr 
by ■-■ ' Cutts and Beer and ;r><:;cie avauar' 

;■, ; . -,d Company, Three of tJaese wo* 

renusSiort, Ihe lor- ■ od being 5 mon,>;i 

definite hii^ -■■i-';iidl- Me ev!d5'^-,ce of : 

sespoj. -.ad in t' 



i-ect?- f;h!!^ course of rr-! '-orna,,; bt 



ii-ain a b ■ 

■.noma, .; ^.i -i- •/■^c.- 

iocarci.' , rheek po^. 



t.hif3 pic; . 

will not: grovy any of the ati-ains of choriocarcmoi-naj and passive 

irjnmunity can foe conferred on othea* hamsters by injection of 

serum froin the resistant animaiSo The resistant state is not 

indticed by injection of sevaral human tiss-ciss including normal 

placenta^ 

The problem of the innnnune relationship between choriocar- 
cinoma and the host subject has been raised frequently because 
this neoplasm is derived from the sygote and contains genes fronn 
both parents^ Dr, Hertz has un.dertaken a study with Dr. Pc, J^ 
Schi-nldt (DBS) of the blood groups of choriocarcinoma patients and 
their husbands „ No evidence of cross -inrimunization between the 
marital partners has been founds even among patients whose tissues 
were extensively invaded by tumoro The scientists infer that the 
ennbryonic neoplasm lacks significant antigenicity for the host, at 
least as far as circuia.ting a.ntibody forrriation is concernedo 

The heterologous choriocarcinoma transp lants produce large 
amounts of chorionic gonadotropin, which are readily detectable 
in the tumoi's plasma and urine but ccLnnot be demonstrated in the 
host's normal organs or tissues. Choriocarcinoma also produces 
the gonadotx-opin in tissue culture o Dr= Hertz finds that neither 
slices nor homogenates of normal hamster organs inactivate the 
hornaoneo Unlike the normal placentas choriocarcinoma displays 
no estrogenics progestational j corticoid,, thyrotropic or adreno- 
tropic acti^rityo 

Dro Hertz is using the transplanted choriocarcinomas in 
screening for chemothes'apeutic activity „ Even though a particular 
tumor may have been taken originally from a patient whose lesion 
was completely resistant to methotrexates the heterologous trans- 
plant still displays moderate sensitivity to the drugo The most 
potent tumor inhibitors found thus far are Vincaleukoblastines noted 
abovcs and a related Vinca alkaloid^ L.eurosine, Other alkaloids 
of Vinca rosea have been inactive when given in tojdc do3es„ 

Thus a program originating in the laboratory has had important 
consequences ir.i the clinic where new and additional problems have 
arisen requiring further laboratory research at an even more 
fundamental leveL It is difficult to study choriocarcinoma in ejcper- 
imental animals a.nd study of the physiology of chorionic gonadotropin 



69» 



preseuts problerr-s. Serum and iirine of the pregnant feraale 
■ monkey, Macacus rhesus , studied by Di',, Hertz^ contaivi detec- 
table amounts of choriomc gonadotropin only from day ZO to day 
35 following timed mating. The hormone cannot be demonstrated 
in concentrates of these body fluids at any other time, nor is it 
found in any of the tissues including the chorion itself. 



Endocrine therapy 

Administration of human pittvitary growth hormone to 10 
patients with metaistatic carcinoma of the breast, three patients 
with cancers of the adrenal cortex and two men with prostatic car- 
cinoma has produced no exacerbation of disease nor other changes in 
theimieaaurable paramieters of tumor activityj according to 
Dr3« Bergenstal and Lipsetto Excretion of urinary calcium is not 
influenced by injection of growth hormone during remission induced 
by hypophysectomy. Only those patients whose cancel's continue tO' 
grow progressively after removal of the pituitai-y respond to growth 
hormone administration by some increase in urinary calcium 
escretiono 

Dr„ Jo M„ Van Btiren (NINDB) has performed section of the 
pituita.ry stalk in il patients with advanced mammary cancers,, 
The morbidity has been greater than in patients submitted to 
hypophysectomys and the oxFerall I'esuits are not as good^ 
DrSo Bergenstal and Lipsett report decline in adrenal cortical 
activity amiong these persons to the levels reached in hypophysec- 
tomized individuaiSp Gonadotropin excretion is variable^ Four 
patients demonstrated norxnal thyroid activity which could be 
suppressed with tri-iodothyronineo The investigators regard 
these findings as evidence for a variable degree of destruction of 
the anterior pituitary following stalk section. Unlike the dog, 
ACTH production seems to be the most sensitive index of this type 
of pituitary damage in mano Thyroid -pituitary relationships in 
several patients have remained noriTsal in the absence of vascular 
connections between the hypothalamus and pituitary, 

Osp' -DDD 



DrSo HertZj Bergenstal and Lipsett have used Ojp' -DDD in 
the treatment of 14 patients v^^ith metastatic carcinoma originating 



■sfe'f-'?*' t^ ■ 



y».i\mt%o: 



ig of charr.otbet 



.^ reapo; 



yOO en: 



.: ■: '• cent <:>\F<i 

particttiariy resrstant to chemotbe:s'apeutic attack, and these 

Carcinoma 7 55 is, the tliti .. .-.cicakjie tiviopiasm to be 

ititroduced into t'ae quantitati'v^e The s'eles'ence standard 



7io 



against which other compounds are tested is o-rnercaptopurineo 
which produces 80 percent increase in sur^dval timet and some 
tumor -free survivors are obtained when treatment is begun on 
the fourth day after inoculation and continued daily for five daySc 
A large number of chemotherapeutic agents will retard the growth 
of this neoplasnn and prolong the lives of the hosts when adminis- 
tered according to the same treatment schedule„ Most of the 
more effective compounds are purine and pyrimidine antimetabolites. 

Much of the research conducted by DrSo Dean Burk and 
Mo W„ Woods is directly concerned with the mechanism of drug 
action„ The writer believes^ however, that the total eicperience 
should also be viewed as a potential means of screening compounds 
for chemotherapeutic activity. The techniques seem capable of 
using climcs-l as v/ell as experimental cancers^ even though some 
technical difficulties may be encountered^ Drs, Woods and Burk 
report that a high degree of malignancy is associated with greatly 
increased glycolytic ca,pacity and lov^ered sensitivity to glycolytic 
inhibition of the anti-insulin type (Annual Eeports, 1956-1958)o 
Generally speaking, those experimental neoplasms in which the 
hexokinase reaction is under sti'ong insulin -anti -insulin control 
respond to one or more chemotherapeutic agents better than do 
those cancers in which glycolysis is less readily inhibited by com- 
pounds of the anti-insulin type^ One does not have to accept the 
theory advanced by Drfi;„ Burk and Woods to recognize that these 
scientists have found a ixeans of classifying ejcperijnental cancers 
with respect both to degree of maligiiancy and potential effective- 
ness of chemotherapeutic drugs„ 

Drs, Burk and Woods have studied the anti glycolytic effect 
of 5-fluorina.ted pyrii^nidines against Krebs-2 and Shrlich ascites 
turaors, finding 5-fluorouridine the most active and 5-fiuorocytosine 
inactivco The in vitro studies correlate closely with the capacity 
of each compound to inhibit tumor growth in vivo » On the other 
hand, a concentration of 400 P<,PoMo of uridine is as active in ^dtro 
as is 20 PoPcMo of 5-fluorouridinee The antiglycolytic activity of 
the 5-fluorinated pyrimidines can be largely counteracted by 
increasing inorganic phosphate in the incubation nnedium, rennoval 
of oxygen or by the use of agents that uncouple phosphorylatiovx. 
The effect measured by Warburg manometry requires glucose; sue- 
cinate^ pyruvate or glutamate ca^i not be substituted for glucose „ 
Exoeriments conducted with Mr» Jo Co Hunter describe the inhibition 



72c 



of aerobic glycolysis o£ rat bone marrow by 5-fiuorouracils but 
not by 5 -flue r ode oxy uridine and reflect the relative degree of 
toKicity of these corrjpoimds for .naice, 

Drs„ Woods and Burk report that the in vitro mechanism of 
action of the 5-fiuorinated pyrimidines appears to involve an inhi- 
bition of aerobic phosphorylation which lov/ers the availability of 
adenosinetriphosphate (ATP) for glycolysis„ Insulin lowers the 
requirement for ATP and should counteract the nnetabolic effects 
of the 5-fiuorina.tsd pyrimidiiies. This it does in S9i rnelanorna 
and normal bone marrow, but not in insulin-insensitive ascites 
tuinors Krebs-2 a.nd Ehrlich, 

Differences in metabolic chE.racteriBtics of tuiTsors respectively 
susceptible and resistant to action of a given chemotherapeutic agent 
are reported by Dr„ Burk and Dr^ K» Mo Wight„ 8-Asaguanine 
prodiices a px'ompt and nnarked increase in glycolysis in vitro of 
susceptible leukemic cells but not of resistant cells with concentra.- 
tions of drug ranging from one -fourth to 4 times the pharmacologica.1 
level commonly used to influence the growth of the sensitive leu- 
kemia in vivOo Respix-atory inhibition occurs in both sensitive and 
resistant cells exposed to S-azaguanine but is seen at lower doses 
with the susceptible line. The metabolic effects characteristic of 
the sensitive cells tend to disappeai' after repeated daily exposure to 
the drug in vivo and may be completely lost v/ithin a transplant 
generation. 

The action of Cytoxan on the metabolism of susceptible mouse 
tunnors leukemia L-lZiO, K-2 ascites and Ehrlich ascites has also 
been s.tudied by Drs„ Burk and Wight, Treatment in vivo with 
removal of the tun^or cells for study in vitro reveals pronounced 
inhibition of aerobic and anaerobic glycolysis as well as of respir- 
ation„ Respiratory and glycolytic functions of a cyto:ij:an -resistant 
sti'ain of LiZlO, prodiiced by Dr. Montague Lane, were usually not 
inhibited by treatment and are sox^stimes stiinulated„ 



Anti-leixkemic compounds 

Cooperative Leukemia Group B in which Drs., Frei and 
Freireich represent the National Cancer institute has completed its 
study of the relative effectiveness of inethotrexate and 6-rriercapto- 
purine given alone or in combination foi* the treatment of acute 



ieukeniiao The stud'jr i^^cludes eatpa rieace with '3Z8 patient.9p o,{ 



whom 92 were ho-.! 
rate was attaineo 
ihe. yate was no I 
duced Vi?ith eithe: 
na:ms for either ^: 
as primary tr°? ' 
..■thsr drug,, 
prodvicad a h-^, 
• i'SKate .; Overar. 
cent for children 
::;.kno3t inactive i. 
effects hi lass tl\ 
"■■Me ifv all group ■ 
■J- rase M": 



rftaa pro- 
.'3 was the 
-.ether nsed 
ition. of the 
alones however,, 
■"fiad with metho" 
;iQS was 50 pes: - 
:;tr<sj(;ate was 
;,g baneficia-l 

compar 

, r • 9 'TO 25 

:'nore orr ' 



the biocheixiical :: 
■.-nental letikeinia-s i-o 
sncountered Irset'ac:^ 
i;aptopurkie la.be 

aietaboli - 












iTrjaianp 



74c 



DCM is the drug most effective against mouse leukemia LiZiO 
and, indeed, some mice are actually cured of their advanced disease 
by use of this compound (Annual Report 1958)« Dr„ Goldin with 
Dro M« A„ Chirigos and MessrSo Venditti, So So Humphreys and 
Go Oo Chapman have conducted extensive experiments on DCM^ 
Daily treatment must be continued 30 to 60 days to permit complete 
recovery froin L,i2i0„ Ordinarily the tumor disappears in about 
tv^o weeks but recurs promptly if treatment is stopped prematurely. 
Oral ad-ministration of both DCM and MTX is less effective in exper- 
imental leukemia than is subcutaneous injection, 

DrSo Goldin, Ao W, Schrecker and J, A. R, Mead (Visiting 
Scientist) report that inhibition of formate incorporation into the 
acid soluble adenine of leukemic spleen is a reasonable measure 
of the effective doses of antifolic compounds given by pa.renteral 
injection^ Maximum effect occurs 20 minutes after injection of the 
drug but one hour after oral dosage^ and larger doses are needed 
to produce the same qua.ntitative effect. 

Some clue to a reason for the increased effectiveness of DCM 
over MTX in the manageixient of experimental leukemia L1210 is 
afforded by other reports from the saine investigators and Dto Ro Ao 
DarroWo DCM has a snT^aller inhibitory effect on forinate incor- 
poration than does MTXo The dose ratio for equal response is 
about tv/Os but the effect from MTX lasts longer. Extending treat- 
ment intervals to six hours or more increases the dose ratio 3f 
DCM to MTX to 25, A single dose of DCM needed to inhibit formate 
incorporation for 24 hours must be of the order of 75 mg/kg, the 
same dose that produces maj-damum survival time of leukeinic mice, 
A comparable dose of MTX cannot be tolerated. 

Mice cured of 1^1210 v^ill usually grow neither a second graft 
of this neoplasm nor cells frorra a subline of L12i0, M46R, which 
resists treatmeiit with antifolic coinpounds, K growth does occur, 
the new tunnor reinains localized to the site of implantationo 
BALB/C mice can be immunized against Li2i0„ This type of 
imnnunity can be overcome, however, by treatrrsent with MTX if the 
mice are inoculated with the antifolic -resistant M46R„ If, on the 
other hand, immunity is induced by injections of spleen fromi strain 
DBA, the natural host of L 12 10, or by other escperimental leulcemias, 
antifolic compounds do not alter the resistant state. 



75„ 



These observations raise questions concerning the role that 
immunity plays in influencing the host -tumor relationship in expei-- 
imentai chemotherapy „ Dx, Goidin and associates have performed 
an experiment in which the markedly resistant subline M46B has 
been affected by administration of a'li antifolic compound. Mice 
bearing both the sensitive form of L,12i0 as well as the resistant 
form M46R were treated with DCM„ Their lives were prolonged 
for a.3 much as 50 days, though treatment of M46S alone with either 
MTX or DCM extends the suz'vival timie to 20 days as co3Tipared with 
the 10 day life ejqsectancy of untreated controls. Presumably the 
effective treatment of the sensitive tumior elicits an imi:nune response 
in the host, which retards the grovi^th of the resistant variant, 

Drs, Goidin. Schrecker and Mea.d report further that large 
doses of MTX itihibit formate incorporation by both sensitive and 
resistant lines of leukemia LlZiO, Inhibition is less pronounced by 
treatment of the resistant cells with small doses of the drug. Such 
differences are quantitative rather than qualitative and are more 
pronounced in the solid tumor than in the leukemic spleen, Citro- 
vorum factor only partially reverses forma.te inhibition induced by 
antifolic therapy. Perhaps the transport of citrovorum factor into 
the ceils may be innpaired by MTX adxriinistra-tionj as was demon- 
strated for certain bacteria by Wood and Hitchings, 

Experiments conducted by Mr, S\ G, Dhyse suggest some 
relatiotiship between other vitamins and folic acid. When biotin is 
added to cultures of L, a.r3.b inosus in excess of the quantity required 
for maximum grovi^th, the cultures give rise to a five -fold increa.se 
in folic acid in the medium without any increase in grov.'th rate. 
The folic acid content of the bacterial cells is not affected, and the 
biotin must be added while the culture is growing actively. If e>:cesj 
quantities of pantothenate or riboflavin, instead of excess biotin are 
added to the medium, e>:cesaive production of folic. acid is not 
observed, 

Dr„ Michael Potter is also interested in folic acid and its 
antimetabolites and uses the tetraploid .lymphocytic neoplasm P288 
rather than .L1210 as a tool, A folic acid deficient diet formulated 
by Dr, G, M„ Briggs (NIAMD) produces better survival of mice- 
beari^lg P288 resistant to antifolic drugs than of mice bearing the 
irug- sensitive counterpart. 



76o 



In studying tliQ acquisition of the resifsfcant stats, Dr, Potter 
inoculates 10 cells into genetically appropriate mice and SO percent 
of the recipients develop leiikemia regardless of whethez* the inoc- 
ulum is derived from ceils resistant or sensitive to the action oS anti- 
folic coiTipoundSo If the sensitive leukemia is treated 5 times within 
10 days with small doses o£ MTX, transplantability falls to 5 percento 
Ceils isolated from P2S8-sensitive lines which haye been treated 
with larger dosas of MTX for longer periods of time grow in 30 to 
60 percent of the mice into v/hich they are transplanted. Further - 
mores the appearance of a. progressively gi-owing mass at the site 
of inocvilation is delayed for some time, suggesting some incoinplete 
and hitherto unknov/n tyg^Q of resistance,, 

Dr„ Goidin, Mr, 'Venditti andDr„ Frei describe a series of 
pyrasolopyrimidines that exhibits a wide range of antileukemic 
activity against L.i210„ The most effective of these chemicals, 
however, is far less potent than either 6-mercaptopurine or metho- 
trexate <, 

Virus "induced leukemsias have been introdviced into the 
chemothsrap'/ prograzrj by Dro Moloney, Dro Goidin and Mr, HuniphreySc 
No increase in survival time has yet been obtained with methotrexate, 
6~mercaptopurine or Cytoxan, even though definite reduction in the 
size of affected organs has been observed. 



Alkylating agents 

A new alkylating agent, cyclophosphamide, developed in 
Germany, is commonly called Cytoxan, Dro Montague Lane reports 
an I.D^^ of 425 mg, for micej, 150 mg, for rats. The drug produces 
little gastrointestinal toxicity a.nd causes hyperplasia of the mega- 
karyocytes in the bone marrow and spleen, Cytoxan is more active 
thaxi nitrogen mustard against leukemia L,1210j including sublines 
resistant to antifolics, lymphom^a L-2, carcinoma 241-6 and the 
Dunning rat le^uikemia and lymphosarcoiriao The drug is most effec- 
tive when administered once weekly, Dr, Goidin has confirjoaed 
Dr„ L-ane's finding v..'ith respect to the action of Cytoxan on LIZ 10 
and reports it at least as effective as methotrexate. Many mice 
survive indefinitely %'yhe.n tresitment is instituted early in the course 
of the transplanted leukemia, Cyto:<an is also, effective against 
adenocarciaoma 755 and S-37, 



77. 



Studies of the toxicology of Cytoxan in dogs and rats have been 
carried out under the direction of Dr„ Do P„ Kali at the Hazleton 
Laboratories, The advantage in terms of administration of large 
quantities of drug through the use of widely spaced dose schedules 
was clearly borne out for these species. Of particular interest is 
the demonstration in rats of a period following adininiatration of 
large doses during v/hich the animal appears to be refractory to 
toxic manifestations of cytoxati or methotrexate « 

Cytoxan in man produces greater depression of leukocytes 
than of platelets according to Dto C„ 0„ Brindley and Dvo Frei^ 
Daily oral doses of 4 mg/kg are tolerated for weeks. Some thera- 
peutic activity is displayed in patients with lymphosarcoma or 
ovarian ca.rciiiomao 

Leuicemia group B is studying the effect of Cytoxan therapy in 
acute leukemia. ■ Drs, Freireich and Frei report no difference in 
toxicity v/hen gi^'-en daily or weekly, but somiev/hat more therapeutic 
benefit appears to be obtained from vi/eekly doses, A total of 96 
patients has been studied of which NCI has contributed 30o Prelim- 
inary results indicate objective improvenrient in 30 percent of the 
patients. 

Uracil mustard, another alkylating agent, has also been studied 
in the clinic, DrSj, Brindley aiid Lane find it active against Hodgkin's 
diseases lyiriphoioaas and chronic leulcemias in oral weekly doses 
of 0,Z mg/kg. The drug depresses the bone marrow and occasion- 
ally causes severe gastrointestinal symptoms. The following oral 
doses are tolerated for siit weeks with minimal toxicity: 

Single dose 0,45 mg 

Daily dose 0, 015 mg/kg 

Weekly dose 0,2 mg/kg 

A comparative study of the effectiveness of thiotepa relative 
to nitrogen mustard in the treatment of Hodgkin's disease indicated 
superiority of the latter drug (Annual Report 1958), Se -examina- 
tion of the data by the Eastern Solid Tuinor Group disclosed some 
evidence that the dose of nitrogen mustard vyas closer to the maxi- 
mun-< tolei'ated dose than was ths^ dose oi thiotepa used in the study, 
Thsrefore,, the investigators undertook s^ study of dose -response 
relationships for both of these alkylatiiig agents. It soon became 



nee therapeutic act' /at when the usvi- 

percent. Whether the ■ , .?r?.pf-:utic effects of thi-jteya anc ruirogen 
n^ustard on Kodgkin".'^ d:ppa«» are ti''-n-)'?.)ar at •■raxTmriliiy tolerated 



During 
, . *■-. wsd the < 
tbonuclease ■ _ 
nccessful, the a.: 
es are also likel 
'-e of serum en^ . 
linr.iied addilior 
icreases in lactir 



■11 preterminal :■ 
for following 



The a.c'fclvr 

■ cer, The 

r.e of cancer 



Protection •: 
irradiation by pri 
brorn.tdehydrobror 
effectiveness vn p 
use c£ nitrogen r; 
ive to nitroge 
• i-eatment will: .■. 
doses, bul: some 
duced by larger c 
pounds which gp'v. - 
protective eff-- - 



r) encouraged 
• bone mart; 



-iTionstr. 



iru.;;oni,ti.;ri 
'vsdy its 

:i with th 



Altho-agh tetracyclines do nC 
<ncer, tfeeij; in:portari-'- ^ntibiotrc 
)ncentrat.e in ce; 



growth c 

ibiHtY to 



79o 



Study of p].asm£ concencrations of tetracycline and demethyl- 
chlortetracyciine after single intravenous doses in dogs revealed 
first-order disappearance curveSo Calculation oi! volumes of dis- 
tribution indicated that demethyl compound and, to a lesser extent^ 
tetracycline are concentrated in some e^rtravascular compartment 
of the bodyo The drugs entered the cerebrospinal fluid in concen- 
trations approxixmating 20 percent of those found in plasma, thouoh 
equilibrium was not attainedo Concentrations in -skeletal muscle 6 
hours after administration were higher than in t'ne plasmao Consid- 
erable amounts of both drugs wers found to be a/jsociated with 
erji^hrocytes. While the disappearance curves from red blood ceils 
were similar to those described for plasma, the presence of dival- 
ent metal ions had important effects which probably influence distri« 
bution of the tetracyclines in ■rivo. 

Formation of complexes of tetracycline with calcium and other 
divalent metals is well known^ Dr, Kohn also describes complexes 
formed with barbiturates and with p-diketones and studies of the 
influence of divalent metal ions on complex formation„ Tetracycline 
forrriS an unionized complex with barbital and either calciunn or zinc 
ionSo Magnesium and manganese fail to form extractable complexeSo 
Pheno- and pentobarbitals are even more potent in causing extrac- 
tion than barbital itselfj and methyl substitution of a single nitrogen 
in the barbiturate nucleus does not change this propertyo If both 
nitrogens are nnethyl substituted, the nnoiecule becomes inerto 
Such complexes do not change either the absorption spectra or opti- 
cal density of the tetracyclines studied, suggesting that the barbi- 
turate may not bind to the chromophoric part of the moleculco 

In the light of these findings. Dr. Kohn has performed equilibrium 
dialysis studies to determine whether tetracyclines bind to nucleic 
acids and proteins„ The most striking results are obtained with 
deoxyribonucleic acids, which bind the drugs in the presence of 
calcium, sine or magnesium ions„ Binding does not occur in the 
absence of divalent metal ions„ Heat denaturation increases the 
binding ability of deoxyribonucleic acids. Ribonucleic acids bind 
tetracyclines less well and albunnin binds to only a small extentc 
The degree of binding is increased in the latter cases by zinc ions 
but not by calciunn or magnesium „ 



80. 



Miscellaneous studie s 

At), intereat in the chemotherapeutic effect of riboflavin 
antagonist, has characterized Dro Lane's activities for the last 
five years because some of these drugs retarded the grov/th of 
certain vex neoplasms but did not prod^ice riboflavin deficiency in 
jnan (Annual Reports 1955s 1956), The Upjohrt Coiiipany cooper- 
ated inost generously in these studies., and sometime ago Merck 
and Company made a different analog of ribo£la\^ins galactoflavin; 
available for this research„ Galactofiavin has produced typical 
riboflavin deficiency in two patients with disseminated cancer, 
and it may now be possible to determine the effect o£ this defi- 
ciency state on the growth of clinical cancerSo 

Lin-iited experience in the treatznent of cancer patients with 
either xnethyl-glyojcol-bis-guanylh^tdrosone or narcotine has not 
elicited any favorable t'espoiissSo 

A compound related to actidione, known as E37, has been 
studied by Dto i?«all in conjunction with the Hazleton laboratories^ 
Female rats are more susceptible So its toxic effects than are 
male ratSj but .ouch sex differences are not found in dogSo This 
drug which Dro Goldin finds effective against leukemia LiZlO pro- 
duces a pecuiitftr hemorrhagic and necrotic lesion in the lungs of 
dogs similar to the lesion described by investigators at the Sloan- 
Kette ring Institute in the lungs of patients who had received this 
a gent o 



SERVICE FUNCTIONS 



Requests for consultation by members of the Surgery Bra.nchf 
NCI, ha.ve incr .eased from ^94 visits in 1957 to 437 visits in 1959 = 
All of these requests are processed through Dr, Smith's office^ 
The gr(3atest change in demand has he^n for urological services v^hich 
has increased 7 tim.es over the 1957 figurcc It is difficult to recruit 
and retsdn a staff of capable surgeons because of the obvious econ- 
omic advartages of private or g;roup practice. 



1 



The .'same considerations apply in an important degree to the 
aliiv c.r cur senior clinical investigatorSc Our pay scale is 

siation to the opportunities that university medical 



Mi e Jueeph Albrecht and his group of eseceilent Mstopatholog^ 
tcbnicians have prcv-ided this staff ^ith s total of 154s 000 stained 
histological preparations of which Z i lUired (special stains 

The a\/erage cost \vas about $ ! -, 12 p«. i , . ..a.. , regardless o£ the- Ria*'-; 
1.?.? .' This isj an unusually low figui'e, especially vyhen one cu-- 
solders ?!■■" ' cost of training unskiiied recruits Is included 

Membai:© o£ the Pathologic Anatomy Branch perforrricd 265 
ii&ciopsies during 1959 of wMch i6i were perforn-ied iox NCL 
They e.xarr.ined 2, 60l surgical specir/ieus and a-ccessloned '^„ 547 
cytodiagnosis specirnenSc Mauy of their ss^sea^-ch roTii.i-'rm,ii!onfi, 
are included in the foregoing nnaterial., 

-J oratory oi' 
'■ ■j-oc'iie.tiiisiij;/ accepfied 5u perceat x».ore speciiiieas tor microch*?:-' . 
A-.niysIs during 1959^ despite a decreas? of 15 percent ina^vailab- 
n.^ji hours,. His prograin on infea~red spectrophotonnetific research 
has ■■;>pex°ated at a reduced ie"*?®!, hut 20 percent of the sarr-^ni*-?; 
aubnusted for empirical analysis are now studied by _infi-r 
tt?chniques fur purposes o£ fstifuc' j .'■ ' i.erpretatii.^n and ay «(i 
indi'i'sition of cheniical purity, 

ihe Aoru'ai Production Section oi the Lab Aids Branch., DBS,, 
■ >nv«jfled to a iee. -for ^service, basis in July, 1V58„ The staiX of 
'~i.-i ■*.?iB therefore required to predict the ne'-- < f-''" n-^''»>, ratfii, 
gais- -a pigSs hamsters and rabbits for an ivx . he predic- 

tion -.> .s <nfed correct within 10 peK'cenfc of actuai us/ij,;'..- ihis in no 
stria I i featf, when one conside-fs the number of persons involved, 
ti«d ■'ss.phasize.a the ability and desire of the Intraroural research 
stall to accept responsibilitys discha rL^e ft n.cst rapeiily and coop-^ 
;i-Ee -Aiiih otheif oyganixationsi segr« ■<» difficult 



. I I'nany «<-'^op[ec ^H»if ci] !e«^T».e® ar^i- <':reati%''-e, The 



J 



82 



to the cancesT patient; yet some ©if it i-equi^es a period of fuir-the;.-' 
development to assay its utility.. We wo«ld prefer to acconxpMsh 
tfeis within tlte framework of a dis-ect or conts'actual operatioa of 
NC4 and hope that we may be eneou^aged to do soo 



Respectfislly submittedt, 





Associate DiFeetOK" im GIviarge- ©i Reseas"; 



Febmary 2. 19^0 



64. 65 

1 :? 



35. bb 
3, 42, 

3. 54 



64 

SB 

i9 
55 

74 
74 
9 

58 



D© .finger, ivL K. 2 3., Z2, 24 

E-'.fa.nrt, V„ Jo '>-^* 



_M I^C, 5y\ ':;■,': 


;■. r. 


'. o I 


Ap.drevy? 


J ^, 


R, 


Barre- 






Base?, 


„ 




Belkin r 






Bergp'.'-/ 






Be rim 






Birnbaui.-: 


1 , 




aiuro, H. 






Brace, K 


c 




Bjfeyere, 


i-,_ 


;; 


Br indie V,; 


c. 




Bryan, Vv 


% K 


D 


Bryant. ' 


r^ c 


o 


B»ark, D. 






C;iaa.pma>i 


, G„ 


o 


Cbirigos, 


, Mo 


A 


^ 'Hways 


Wo 


D„ 



50, 






50, 


li-. 


77 


•> '7 


70, 


74, 


U] 


S4 




z, 


3s y 


h 38 



Go Id? :•:!., A<, 
Greetiiield " 
Gteensleir. 

Haxrit,- ■-. . 30 

Hertz, Roy •. 
Heston, Wo S, 
Hilberg, A.- V/„ 

Hoilcroit, J, 46; o.i 

Hoye, R, C, 3i 

Hueper, W 4„ 6, 7, 8 

Hiirtiphrev - 74, 76 

Hunter 71 

Izvtmiyc'. r' 36 

Kahler, H„ i -9 

Kaisex, R. F. S 

Kelly, M. 6, vs 

Ketcharris A„ S, 
Kielley, Ho K, 

Kinsey, D, L. 30 

Koegel, Re J„ 8i 

Kohn, Ko W. 7S- 79 

Kuff, E. L. 
K'ajfatsune. M„ 

jLaady, M„ 

Lane, Mo ^ 

LaWf i. '^' 

Letter,, j. 

Levin., J,, 04 

Li, M. C. 67 

Lipsett, M„ Bo 3, Al, 43, 6-;/ 

Lockhart, Ro 2,., 47 

Loo, T. Lo 50. 75, ?' 

Lot^es Bo 14, 56, t 



Mii'ts2.xid.GV . 
Mxchi, K,_ 
^4oioney;, J, 



F' otter 



JRabeon,, A.. 
Rail, D. P 



Ro&fenbe.i'g; 






40, 4.1 
30 



Smith, 
Smith, 

Smith, 



n, 43 
48,: ';'-^ 



JOG-;; 

iitaeheiliT; 
5tantor. '■ 
Stewart 

^ugirr.ura, 

Huston.,, H, 
a v; ami J P, , 






Mc'.tth'tas 



i.. 



6^^ 

64 
?-0„ 65 



Ihomaa,. i.,, 5,, 
Toal, J. N. 
Taicfeudy, D„ P» 



5> 

65 



lipfcsolf. Do £, 
Van Scoit, Eo J. 



57. 58 
70. 74. 76 



Watkitis Do M, 

Weksburgerj J^ fJ,. 
Westfaii, Bo B„ 
We.>sler8 Hilda 



10 

10, 39 
54 

/i9 



Wimtz... 
V/oilms? 
Wood- .? 



VotiJsa, B; 



^ubr ■■'•[ 



b 



lendar Year 1 



TABLE OF COIWENTS Pages 

OFFICE OF THE DIRECTOR 1-3 

Office of Information and Publication k •> 9 

Journal of the Hational Cancer Institute 10 

BIOMETRY BRMCH 

Summaiy Report 11 - lU 

Project Reports s 

lICI-50 Acti-\d.ties during 1959 15 » l6 

WCI-52 Cancer Morbidity Surveys in the lM.ted States 17 » 19 

NCI-53 Statistical Sttjdies of Cancer in Pittsburgh Area 20 - 21 

NCI-5^ Studies on Cancer in Connecticut 22-25 

KCI-55 Retrospective Study on the Association of Smoking 26 - 28 

and Lung Cancer in Women 

HCI-56 Association of Smoking History j, Residence History^ 29 - 31 

and Occupation History with Lvmg Caacer Mortality 

KCI-57 Studies of cancer mortality data 32 - 3^- 

KCI-58 Constiltation on cancer registers and application of 35 "■39 

statistical methods to study of csxicer in human 

populations 

NCl-51 Statistical Consulting and Resiearch Services ko - k3 

CANCER CHEMOTEERAPI KATIOKAL SERTOCE CENTER kk - 73 

FIELD lUVESTISATIOTS AKD DEMC1ISTRATI0N,S BRANCH 

Summary Report 7^ = 81 

Project Reports s 

Hex-Si Siiperyislon and administration of research pro» 82 - 85 

jects investigating exfoliative cytologic 

methods^ etc. 
WCI=.83 Early detection of carcinoma of the colon by 

cytological and chemical techniques. 
NCI~8l|- A study in a general population of cervical cancer 

using vaginal and cervical cytology for prelimi=" 

nary case selection = Columbus, Ohio 
NCI-=85 Peripheral Blood Project = Winston^Salem, K.C. ^ 
NCI-I05 Cortisone Study=.Georgetown University Medical 

School and Georgetown University Hospital 



36 . 


- 90 


91 ' 


^9k 


95 ' 
98^ 


- 97 

=■ 99 



M:I»86 


NCI-87 


KCI«88 


NGI-89 


ICI-106 


KCI'^90 



Investigation and Development of tfethods of 

Collection of Sputum Specimens-Houston, Texas 
Investigations and Development of ^fetllods of 

Preservation^ Processing, and Staining of 

Bronchial Secretions"-Ho\iston, Texas 
Histologic and Histopathologic Study of Sx" lOU ■ 

foliated Bronchial CeilS"»Houston,, Texas 
A study in a general popiolation of cervical I06 j.:/;, 

cancer using vaginal and cervical cytology 

for preliminary case selection^Loulsvilley 

Kentucky 
A uterine cei^acal cancer morbidity study in HO - '"'"' 

Louisville^ Kentucky 
A study in a general population of cervical 113 ■=• -u -. 

cancer using vaginal cytology for prelimi= 

nary case selection =■ Madisonp Wisconsin 
KCI-91 A retrospective study in a selective patient 115 - 13.6 

group of cervical cancer using vaginal cyto- 
logy for preliminary case selection— ^Sadisons 

Wisconsin 
KGI'=.9U Field evaluation of the performance of an elec«= II7 = II9 

tronic cytoanalyzer . Investigation of new . 

methods of preparing -vaginal cervical, material 

for automatic scanaalng. Hagerstown^ Maryland 
Examination of sputum specimens from uranium 

miners. Investigations to eralixate cytologic 

morphology » etc. 
A study in an industrial population of cervical 121 - x?.^- 

ccmcer using vaginal cytology for preliminary 

case selection->Fhiladelphiaj, Pennsylvania 
A study in a general population of cervical 125 

cancer using cervical cytology for preliminary 

ease selection^San Diego, California 
A detailed investigation of Gastric Cancer utiliz- 128 - 130 

irtg Exfoliative Cytology for case selection^ 

w'inBton=.Salem, N. C 
Studies on distribution of exfoliated cancer 131 - 132 

cells within tracheo-bronchial tree, correla- 
tion with the cellular morphology of the same 

tuinor from imprint smears, etc . -Houston, Texas 
Dia(^o3tic studies of gastric carcinoma 133 •= 13^ 

Fluorescent microscopy studies in neoplastic di- 135 -^ 137 

seases and in conditions manifesting ejati= 

nuclear factors = Hew Haven, Connecticut 
lCI-110 Orotidylllc Acid 33ecarboxylase Activity in the I38 

Peripheral Blood and Tissues of Patients with 

5fe;.'plastic Disease and Normals- 



II 



iici»i03 


MCI-95 


KCI=96 


mi^i6k 


miliar 


NCI==108 


KCI=109 



TABLE OF CCMEENTS 

Field Investigations and Demonstrations Branch (Cont'd.) 

KCI»111 Leukemia Occurrence in the New England States 139 - l4l 
NCI=112 Followup of Children Treated by X=.ray for . 142 =■ IU3 

Lyisiphoid Hyperplasia 
ITCI==113 A Study of Etiological Factors in Leukemia Ikk « ihS 

and Other Malignancies of Childhood 
NCI"ll6 Geographic Distribution of Respiratory Cancer lit-7 » lk6 

Mortality in I5 Selected States. 
SJCI«117 Possible Effects of Atomic Pall-out as Re= li».9 = I50 

fleeted in Vital Statistics 
NCI»119 Geographic Distribution of Cancer in the U.S. 15I = 153 
KCI»120 Geographical Incidence of Cancer of Major Sites 154 - I55 

in IJpstate New York 
1K;I=.121 Cancer as a Cause of Death and Disability 156 - I57 

Among Workers in the Railroad Industry 
1ICI<»125 Types of Cancer Among Railroad Engineers^ I58 - I59 

Firemen^, and Trainmen 
KCI-I26 Geographic and Environmental Factors in Cancer 160 <=- 164 

Mortality in Washington County^ Maryland 
NCI=127 Development of Methods for Mgasuring External I65 = I66 

Radiation Exposiire in Mines as Related to 

Cancer Occtorrence of Uranium Miners. 
NCI"128 Study of the Cancer Experience of non-Uranium I67 = I68 

Hard Rock Miners in the Rocky Motintain Area 
HCI-I29 Cancer Occurrence in Uranium Miners of the I69 « I72 

Colorado Plateau 
Age Determination by Means of Hair 173 

Effect of SeleMum Sulfide on Hair Roots 174 = 175 

Study of Occupational Backgrounds of Hospitalized I76 = I77 

Cases of Liaig and Bowel Cancer^ Leukemias, and 

Primary Malignant Tumors of Bone in Males Over 

the Age of 25 Years 
Irradiation Induced Tumors in Children I78 = 179 

A study of cancer of the uterine cervix in the I80 = I83 

Jferaphis=Sbielby County population 
ITCI"170 Research in epidemiologic toethod, stimulation l84 = I86 

of epidemiologic research in non=Service 

agencies^ and development of sources of 

Pleld data related to specific epidemiologic 

questions 
KCI>=171 Cancer in a closed population I87 « I88 

NCI'='172 Geographic, Racial^ and Other Factors in Distri» I89 = 190 

bution of Cancer 
NOI<=173 Epidemiology of cancer of the Ivrng^, Pittsburg I9I =. 193 

Field Study 
IICI-139 Relation of Smog to Lung Cancer I94 =. I95 

NCI"l40 To further test the finding of an increased risk I96 - I97 

of leukemia subseq\ient to infection with 

chicl!:en"pox 

m 



KCI-129 

(suh 1) 

KCI-129 

(sub 2} 
NCI«130 


NCI==131 
HCI-92 



Field Investigations and DemonstrationB Braach (Cont'd.) 

NCI-l^H Relationship of Cancer Occurrence and Socio- 198 ■=• 199 

economic Factors 
mi-lk-Z Evaluation of Death Certificate Reliability 200 - 201 

as Soturce of EnTironmental Cancer Informa- 
tion 
HCI-155 Occupation and Lung Cancer 202 - 204 

NCI-l41v Clinical Trials TJnit 205-208 

NCI-lij-5 A study of lymphoma cases treated in a selected 209 - 210 

hospital in New England, including observa- 
tions on patients and records 
HCI-146 Cancer incidence in a population of 5j(CX>0 211 - 212 

persons examined periodically 
W^I-lkj Cancer of breast as related to lactation, 213 °- 23 

pregnancies and hormone therapy 
mCI-1^ Cancer Mortality among Catholic Brothers and 215 ■» 2l6 

Sisters in Several Tteaehing Orders in the 

United States 
NCI-168 mtraTiolet Microscopy as an Aid in Detecting 21? - 218 

Malignant Disease 
NCI-149 ^persensitivity and other host tissue factors 219 "• 220 

in cancer of the stomach and large bowel 
IK2I-150 Oral cancer in relation to vitamin deficiency, 221 - 2S2 

use of snuff, and other factors 
KCI-I52 Correlation of thyroid disease and breast cancer 223 - 224 
HCI-I53 Caneer mortality in several selected population 225 - 22? 

groups in Ne^r England 
KI-154 Excess Lung Cancer among Mexican Females 228 - 230 

HGI-157 Achlorhydria Survey of the Japanese Population 231 - 23U 

of Los Angeles by the fUbeless Gastric 

Analysis Met^sod 
NCI-164 Circumcision and Cervical Cancer 235 - 238 

NCI-159 Study of about 100 cases of polycythemia vera 239 - Si^C' 

treated with p3^ 
KCI-I60 Frequency of occurrence of cancer and other 24l - 242 

major diseases among a grotcp covered by a 
8 state-wide unemployment insurance 
HCI»l6l Gastric secretory function in selected popiila- 2^3 - 246 

tion groups and evaluation of tubelese 

gastric analysis, plasma pepsinogen determina^ 

tion and Schillings tests as screening tests 

for gastric cancer. 



sld Investigations and Demonstrations Branch (Cont'd.) 

IICI~l63 Spectrophotometric analysis of leukemic 

white ceHs. 
NCI'=l65 E3.ectrocardiographic Chenges Associated with 

2 Million Volt X-Ray Therapy to the Chest. 
HCI«l66 The relationship between carcinoma of the 

endometri'uin and diabetes tnellitus. 
NCI-I67 Amino Acid Determinations of White Blood 

Cells, Using Paper Chromatography 
KCI"199 Grants Section » Summary report 
HCI«l82 Caacer Mursing Bibliography 
MCI"l83 Preparation of a Monograph on Radiation 

(For Nurses ). 
UCI=l8i|- Investigation of Problems (Mental and 

Physical) Faced by Patients Ondergoing 

Treatment for Cancer at Roswell Park 

tfemorial Institute. 
NCI"l88 VJork conferences (on Nurse Patient Rela- 
tions arid Problem Solving Tests) through 

State Leagues for Hursing. 
IICI-I93 Appointmeat to Committee on the "Role and 

TrainiBig" of Professional Personnel-.™ 

White Eoixse Conference on Aging— 1961. 
3!JCI=»19l!- Attitudes Test (About Cancer Nursing) 
!iICI»195 Study of Ifethodology for Conducting Mursing 

Resear-ch Pi'ojects. 
I!ICI»196 An exploration of the feasibility of meiking 276 

a comparison of the incidence of cancer 

and. other diseases in a selected group 

with the incidence of those same diseases 

ini the general population. 
MCI-I9'" ConBiultation to laiiversities interested in 277 

b'/wecialized programs in Cancer Nursing 

((Baccalaiireate and graduate progrsoms ) . 



RESEARCH GRANTS! BRANCH 278 = 30'+ 



2i^7 - 


- 2k9 


250 . 


» 251 


252 . 


- 253 


25it ■ 


- 255 


256 . 
266 . 
268 . 


» 265 
=. 267 
^ 269 




270 


271 


■- 272 




273 




27^- 
275 



IMTRAMURAL RESEARCH EROSEAiyB 

Office of the Associate Director in Charge of Research 

Project Reports; 

NCI-15-OOO Ce)Screening 305 

HCI-15-001 (cjservice for other Institutes 306 

NGI-15-002 (c)BBergency Admission© 307 

NCI-201 Studies of Gytochemical Changes in Living 308-316 

Individual Cells 
KCI-216 Drug Development and Evaluation Program 317-335 

KCI-203 Ifecromolecular Polymers as Carcinogenic Agents 336-3^^ 
KCI-.20i^ Carcinogenic Studies on Petroleum Substitutes ^^^-^^-^ 

and Synthetic Liquid Fuels 
NCI-205 Experimental Studies in J&tal Cancerigenesis 3^9-368 
NCI-206 Carcinpgenic Studies on Food Additives and 3^9-39^ 

Contaminants 
NGI-209 Jfedicolegal, Consultative, Advisory and 395-398 

Educational Activities in Environmental 

Cancer 
KCI-210 Studies on Occupational and Environmental 399„li.l3 

Respiratoiy Cancer Hazards^ etc, 
KCI-211 Epidemiological Studies of Environmental 4li|.=,24.15 

Carcinogens 
HCI"212 Experimental Carcinogenesis of the IJpper i4-l6-U21 

Digestive Tract 

Laboratory of Biochemistry 
Project Reports s 

HCI=301 Secretary Duties in the Laboratory of J+22 

Biochemistry 
NCI-302 Microchemical empirical analysisi Develop- ii-23-42^ 

ment of microchemical methods | Infrared 

and ultraviolet absorption spectroscopyo 
KCI-306 Studies on Hydroxyamino Acids k2^-k2J 

NCI-313 Chromatographic Analysis of Pax>tein Com- iv28-U3l 

ponents of Serum in Health and Disease = 
1ICI»335 Quantitative Nutritional Studies with Vfeter- 432-it-38 

soluble^ ChemicaJJLy "Defined Diets 
NC1=339 The Fractionation of Oligonucleotides Derived 439-441. 

by Enzymic Degradation of nucleic Acids 
UC1=356 General techniques end concepts for the 442-445 

chromatography of macromolecules 

VI 



h62 



Homogeneity sno. re of Proteins kkS-hUQ 

iHOl-'^oi. Studies on the D-Peptidase Activity of Hog Ui^■9-i^50 

Renal Aminopeptidase . , , 

ISCl-S^S Separation of Diastereomeric a "Amino Acids by 451-4!>2 

Column Chromatography on a iyJacro Scale . 
lCI-365 Investigation of Cjriioplasmic Hucleoproteing 453-454 

and Their Role in Protein Synthesis. 
liCI"366 Structural and Biochemical Investigations ^55-457 

of Intracelltilar Corapoiaentf; . 
MC1=330 CtMibined Actions of Anti»Cancer Agents and it58-46l 

Hormonal Systems that inhibit Cancer Mata- 

holism and Growiih. 
HCI=331 Subcellular localization of enzyme,® involved 

in the carbohydrate metabolism and growth of 

tumors . )£.)(: 

Krc-332 Effects of Steroid Hormone Balance on Growth hb-^-kb^ 

of Ttansplajated Mbufse Tumors. 

SICI-333 Fundamental Oxidative and Glycolytic Meta= U66-'+70 

holism in tuaors aiad Tissue Cultures. 
WCI-33^ ^e occurrence and fuiiction of a etresE" 471-474 

modifiable anti=.insuliii hormonal complex 

regulating glucose utiMzation in cancer 

metabolism and growth. 
lCI-320 Studies on the rate.® and kinetics of enzyme 475-480 

synthesis and destiruction in vivo. 
NCI=34l Studies on iron metaboliem in the tumor= ii6l-484 

bearing host . 
3JCI-353 The Chromatographic /malysis of the Ribo- iv85-487 

nuclease Activities in jfJormal and Neoplastic 

Tissues. 
101=359 Studies on the fracltionation of erythrocytes 488-=492 

and other tissue cells in normal and tumor^ 

bearing animals . 
IICI=360 Phosphodiesterases of noraal and neoplastic 1+93.494 

tissues . 
HCI=.363 Studies on the Cachexia of Tumor ^b earing 495-497 

Animalg. 
HCl-36it- Tils DNase activities of normal and neoplastic i+98-499 

tissues 
KI"32lt Biological studies of the mechanism((s) of 500-503 

carcinogenesis 
ICI-B25 Some Biochemical Aspects of Chemical Carcino=. 506-514 

gens^s etc. 
KCI-327 Ifechanism of Carc^inogenesis - Metabolism of 515-520 

S=2"Fluorenylacceta!i!ide euod Related Compounds. 
HCI=328 Role of iodine, thyroxine, thyrotrophin, axid 521-523 

goitrogens in Ifehe development and growth 

of thj'T'oid turjors in the mouse and the rat. 



\'II 



TABLE OF CO03TENTS 
laboratory of Biochemistry (Cont'd. 2 

NCI-.346 & 3i)-9 Influence of liver fimction in rats 52^-525 

receiving a ehemical hepato-carcinogen 
and variations of B5 intake as related 
to the development of liver neoplasms. 

NCl-3i|-7 Role of Tryptophan and Indole in the Indue- 526-527 
tion of Blaxider Tumors in Rats Ingesting 
N=2-fluorenylacetamide (2='FM). 

HCI=3l4-8 Influence of the thyroid gland in the develop- 528-529 
ment of liver cancer in the rat ingesting 
M-2»fluorenylacetamide or ili[=>2-fluorenyl= 
diacetamide . 

Laboratory of Biology 

HCI-li-OO Biological studies on the factors involved 530-5^ 

in the development and growth of tumors 

in experimental animals . 
NCI«lt33 Studies on the viral etiology of neoplasmg 5^1-5^7 
NCI"i{OJ+ DM synthesis in normal and neoplastic 5^-550 

tissiies and cells. Synthesis of phos=» 

phorylated DM precursors. 
HCI"ii-06 Electron microscopy of normal and malignant 551-55^ 

cells, including studies on viruses . 
"ECl^hOJ Deoxyrihose-containing compounds in normalj, 555''56o 

regenerating and cancer tissues . 
UCI=l408 Studies on lymphocytic tumors 561-565 

ITCI^if-C^ Electron microscope studies on various norraaly 566-568 

virus associated and malignant cells. 
HCI-ij-lij- Diffusion chamber and transparent chamber 569-572 

research 
KCI<=i<-17 Investigation of the role of genes and their 573'=578 

relationship to non-genetic factors in the 

development of cancer. 
KCI-418 Host-Tumor Relationships 579-58^+ 

NCI-il-21 Genetic studies in mice. A. Tumors in mice. 585"591 
NCI-J4-27 Studies on the etiology and therapy of experi- 592-6o6 

mental lymphomas 
SICI=Jf27(a) Studies of the etiology ^^ pathogenesis and 607-613 

chemotherapy of reticular neoplasms in 

mice 
HCI=i»-27(h) Studies on Biochemical mechanisms of sensi- 61J4-617 

tivity and resistance to antimetabolites. 
NCI=i<-27(c) Factors effecting transplantation of hemato- 618-628 

poietic tissue in the irradiated mouse. 
JTCI-i(-27Cd} A study of the etiolo^'' and chemotherapy of 629-634 

experimental lymphomas 
NCI-i4-28(a) Microcinematographic studies of tissue cells 635=637 

in vitro. 



VIIL 



Project 1 
Project 2 



nci=U28(c) 
Project 1 

Project 2 

NCI"428(c} 

Project 3 

NCI»i^28(d} 

NCI=428Ce)) 

Project 1 

UCI<=I^28Ce} 
Project 2 



IICI-1^28Ce} 
Project 3 

HCl='^28(e) 
Project 4 

HCI=l»28(f) 



WCI=l4.10(a) 
NCI=.l4-l6 



Rapidly agitated fluid suspension c\iltures 638"6^0 
for the gi'owth of cells of various esta^ 
"blished cell strains . 

The growth-potentiating effect on chemically 641-6^4-5 
defined medium NCTC 109 of ifethocel with 
certain cell strains in a} rapidly agitated 
fluid suspension cult-ures and b) small 
static culttires. 

Studies of Cell lutrition in vitro 646-651 

Cell transformations in vitro. 652-657 

Collah orations in Tissue Culture with other 658-662 

organizations . 
Studies of Cell Metabolism in vitro. 663-666 

Vitamin requirements, of a strain of mouse 667-668 

cells as tested in a protein=.free chemically 

defined medi-um. 
Sarcomatous transformation and persis<= 669-67O 

tence of the mammary tumor agent in 

long«term cultures of mouse mammary 

carcinoma . 
Cell transformations in vitro 671-673 

Some effects of pc-lyoma infection on cells 67^-675 
in long" term tissue culture. 

Studies in tissue culture of the malignant 676-68O 
cell and of the normal cell from which 
it arose. (This is a gene^cil title for 
all work in this Section) . 

Biological', and biochemical studies on a murine 68I-687 
leukemia virus recovered from Sarcoma 37- 

Collaborative biologicalj, biochemical and 688-691 
physical studies on a mouse leukemia virus 
recovered from Sarcoma 37- 

Application of immunological techniques to 692-698 
the problem of demonstrating viruses in 
hvonan and other tumors, either directly or 
by revealing the presence of specific 
antibody. 

Viral 'Therapy of Tumors. I. Adaptive Msasures 699-700 
for Increased Gacolysis in Cases of Ad« 
vanced Carcinoma of the Cervix. 

The Application of Tissue Culture to the Study 701-702 
of Tumor Viruses. I. Effect of Mouse Tissue 
Culture Material Previously Treated with 
Human Leukemic Brain Extracts on Tumor In- 
cidence in Swiss mice 



IX 



TABLE. OF COIStTENTS 

Jooratory of Biology (Cont'd.) 



Pages 



NCI»4l6(bj The Application of Tissue Ciilture to the 103~70k 

Study of Tumor Viruses. II. Viruses Aseo= 

ciated with Human Tumors. 
HCI~^l6(c} The application of tissue cvilture to the 705-708 

study of tumor viruses . Ill . Maintenance 

of a lei:ikemia virus of mice in vitro 
NCI«l<-20(c) Studies on the Rous sarcoma virus. III. Fur- 709 

ther studies on the relation of recoverable 

virus to tumor initiating dose. 
NC-I"teO(d) Studies on Rous sarcoma virus. IV. Investi= 710-712 

gations on host response to the virus (a)in 

chickens and (b) in Japanese quail 
KCI«420(f } Search for tumor inducing vijruses among 713-715 

maimnalian tumors (including human) not 

now kno^iTQ to have a viral etiology 

Laboratory of Chemical Phaannacology 

NCI-'250 Planning, Management and Coordination of 716-723 
Investigations, Organized in an Inter = 
disciplinary Team Approach, on Basic Sci'^ 
ence Aspects of Pharmacology in Cancer. 

HCI-253 BioclieBiical and Pharmacological Studies of Host- 724-739 
Tumor Relationships in Tumor Chemotherapy. 

NCI='252 Pharmacologic Action of Tumor "Damaging and •jkO-'Jh^ 
other Chemical Agents on Animal and 
Human ffelignant and Mormal Cells. 

]!JCI=262 Biophysical study of macromolecular interaction 75O-762 
in the inhibition of enzymes and other bio- 
logic ally™ active macromolecules . 

IICI-2SI The role of polysaccharide coTrrplexes in re- 763-775 
sistance of the host to injurious influences . 

Endocrinology Branch 

lCI«800(c} Endocrine Aspects of the Progression and 776-779 

Therapy of Cancer of the Breast in Women 

and Men. 
NCI->803(c) The Nature of Hormone=Producing Tumors of 78O-787 

Pituitaiy, Adrenal ^ Ovary, Testis i, Pancreas, 

Parathyroid and Chorion; Abnormalities of 

Somatic Development and Grovth. 
NCI"8o8(c) Endocrine Factors Governing Hormone "Induced 788-793 

Tissue Growth and Tumor Formation in 

Animals I Bioassay of Blood and Urine 

from Cancer Patients 



TABIE OF COOTENTS 
Endocrinology Branch (Cont'do) Pages 

NGI-810 Phanaacological and Physiological Studies on 79^-799 
Alteration of Endocrine Function in Animals, 

HCI-813 The Preparation of Crude Katural Exfcracts 800-802 
of Hormonal Activity from Urine and Serum 
and their Purification and Physical-Chemical 
Chaaracterizations „ 

lTCI-814 The Preparation and Evaluation of Biotin Anta- 803-80U 
gonists in the Study of Biotin Mstaholism 
as en Approach to Cancer <. 

General Ifedicine Branch 

jICI»i|-709Cc) The Chemotherapy of Acute Leukemiao 805-808 

UCI-I(-~715(e) Ihe ^feehaaism and Management of the Hemorrhagic 809-81I 

Disorder Associated with Acute leiiicemiao 
IIC:r»U»719(c) A Study of the Clinical Manifestation of Acute 812-814 

leiiiseraiao 
Hci=»4=73l4-(c) Assay of Erythropoietin in Bone Marrow Sus- 815-816 

pensions - 
NCI'=l^-735Cc) Oaie Effect of Therapy on the Synthesis of 817-818 

Desoxyribonucleic Acid by I^eukemic Cells o 
KCI»ij-"736Cc) The Demonstration of a Hoiroone Governing the 819-820 

Production of Platelet©^ 
UCI-5-712Cc) Ao Comparative Trials of Chemotherapeutic 821-826 
Agents in Patients with Solid Tumors <. 

Ba Initial Therapeutic and Pharmacological 
Studies of Chemotherapeutic Agents in 
Patient© with Solid Tumors o 

HCI-5-737{c} Infection^, Fever and Host Resistance in 827-830 

MsJJ-gQant Neoplastic Disease » 
KCI-l"2£3(c) Penetration of Anti -tumor Dr\igs into the 83I-833 

Central Nervous System 
MGI-l-225{e} The Pharmacology of Antitumor Agents Related 834-835 

to Purine. 
KCI"i-226(c) The Jfetaholism of Folic Acid Antagonists „ 836-837 
WCI-.I-227 i^ehanism of Antitumor Action of Purine 838-840 

Ana-logs 
HCX-l"'229Cc} Pharmacology of Riboflavin Analogue 841-843 

K:t-l=.23l(e) Studies on Tetracycline© 844-848 

KCI-1-232 Pharmacology of Drugs Influencing the Response 849-851 

to Radiomimetic Drugs and Radiation 
IICI-l-233 Chemical Carcinogenesis 852-854 

IICI-l'-234(c) Pharmacology of Alkylating Agents 855-658 

NCI-l-235 Preclinical Toxicology of Drugs Potentially 859-861 

Useful in Cancer Cbemotherapy 



XI 



TABIE OF CCTOESHTS 
General Medicine Branch (Cont'd.) 



Pages 



NCI-1-236 Effect of the Hypermetabolic State in 862 

Malignancy on I&tabolism of Tbyroxine 

and Hydrocortisone . 
NCI~l-237 Biochemical and Pharmacological Studies 863-86U 

of Human Neoplasms in Tissue Cultiores 
NCI-7-708(c) Clinical Investigations of Ifelignant Skin 865.-868 

Diseases and of Benign Conditions Related 

Thereto . 
ICI-7-T28(c) Struct\iral and Physiological Characteristics 869-871 

of Normal and Pathological Epidermis and 

Hair Roots 
NCI-7"730(c) The Biochemistry of lormal and Pathological 872-875 

Epidermis . 
NCI»6-702(c) Serum Proteins Studies 876-879 

NCI-6=.70U(c) Studies of Porphyrin ifetaholism in the 88O-881 

Tumor Bearing Host and Porphyria. 
KCI=6=.72l(c) Metabolism of Nutrients and Calories in 882-887 

Neoplastic Disease 
KCI-.6-723(c) Studies of Human Serum Albumin Labelled 888-892 

with 1I3I 
NCI=-6.725(c) Vitamin Bx2 metabolism in normal subjects 893-896 

and patients with neoplastic and other 

diseases. 
NCI-6»726(c) Renal clearance studies in patients with 897-898 

neoplastic disease. 
NCI=6-727(c) Gross Body Composition 899-90I 

NCI==6c=729Cc} Studies of Nitrogen Metabolism using NlJ. 902-90^+ 
NCI«6-732(c} Investigation of Malignant Reticular Disease 905-907 

and Serum Protein Abnormalities. 
NCI-6~738(c) A Study of the Factors Controlling Red Blood 908-910 

Ce3J. Synthesis and Destruction. 

Pathologic Anatomy Branch 

NCI=852 Exfoliative Cytology Applied to Human Diag- 91I-915 

nostic Problems and Research Problems 
NCI-853 SiH-gical Pathology and Postmortem Service 916-924 

Laboratory of Pathology 

NCI-500 Histogenesis and Pathology of Induced and 925-928 

Spontaneous Tumors of Laboratory Animals 
NCI-502 Histogenesis and histopathology of 929-931 

cutaneous neoplasms . 
KCI-525 Attempt to induct carcinoma of the bladder 932-933 

in hamsters by infection with Schistosoma 

haematobi-um . 



XII 



aboratory of Pathology (C'. . 

IK;I=528 Cjrkochemistry of Ribonuc3.eoproteiiifs 

NCI-529 Viral Therapy of Tumors 93S=9"<' 

KCI-530 St^idy of Filtrable Agents from Mouse '?' r=9't3 

Tumors o 
KCl-536 A review of the ©urgical suad autopsy 8ec° 

tions from Veterans of World War I whose 

caxise of death has been listed as lung 

cancer 
KCI-536{a) Eacperimental studies relating to the jnini= 9^5-9^7 

mal concentration of hydrocarbons which 

will produce cancer in hamster cheek 

pouches o 
NSl-JOS Collaborative Research Requiring Pathologic 9^=9^9 

Anatouy 
NCI=50^ An Electron Micro^cc^ic Investigation of 950-951 

Hormal and Heoplastie Tissues.-. 
mi'-36kia.} An Investigation of the Coilagenous Con- 952-953 

nective Tissues and Elastin, 
MCI=505 The Induction of Squamous Carcinoma of the 95^=-955 

Lungs in Laboratory Animals 
WCX=506 study of Ti-ansplanted Tumors in Mice 956-=957 

1K;I°507 Factors Involved in Abortive Jtttosis in 95^ 

Human Hair Follicle® in the Catagen (rest- 
ing) Haase of the Growth Cycle 
KCi°508 Cytology of Antibody ^producing Plasjna»cell 959-96C 

Tumors of Mice and Hvmsan Multiple Myeloma 
KCI-509 The ProtqplasiD of Tumor Cells s Characteristics 9^1 

of maligoant neoplastic protoplasm 
l?CI=510 Study of Sodocrine and Reproductive Organs 962=961+ 

in the Mouse 
NCI-511 Collaborative Research 965-966 

I}Ca=512 Transplantation Studies with Particular Em= 967-969 

pihasis on the ^fo^phologiC£^ Alterations 

of Cartilage Tissue and Endochondral 

Bone Formation 
15C3;-5i3 Ontogenetic Development of Plasma CeHs in 970-971 

Mice 
HCI°5l4 Induced and Spontaneovis Neoplasms in Labora= 972-97^ 

tory Animals a 
KC;C=.515 Effect of the Growth of Adrenocortical 97^ 

Carcinoma on the Chemistry and Physiology 

of the Host Rat, 
Wa:C-5l6 3t\idies on the Induction of Multiple Tumors 976-978 

in Certain Mammals with Stewart-Eddy 

Polyoma Virus-tiEstie Culture Preparations 



XIII 



TABLE OF COMEEKTS 
Laboratoiy of Pathology (Cont'do) Pages 

KCI-518 Experimental Induction of Adenocarcl- 979-981 

noma of the Glandular Stomach in the 

Moxise 
HCI-519 Corticosteroid Metabolism of an Adreno- 982-983 

cortical Adenocarcinoma of Rat, 
NCI-520 Moiphologic Study of the Effects of 98U-986 

Radiation Protective Conipovmds 
KCI-521 Morphologic and Biologie StMies of 987-989 

Leukemias amd lymphomas of the Mouse 

in Tissue Culttsre 
WCI-"52l{a) Tissue Cultiire Studies Investigating the 99O-996 

Mschanism of Tumor Induction by Parotid 
• Tumor Agent (Polyoma Virus) 
KC1=>522 Studies on the Induction and Natxiral His- 997-998 

tory of Conjunctival Carcinoma in Here~ 

ford Cattle 
NCI~523 Effects of Ionising Radiation with Enrphasis 999-1001 

on !43rphologic Alterations and Neoplasms 
HCI-^^l}.! Pathologic Findings in Human Trophoblastic 1002 

Disease o 
NCI-526 Chemical Carcinogenesis IOO3-IOOU 

NCI-526a Chemical Carcinogenesis IOO5-IOO6 

NCI-527 A Study of the Ehysiopathologic Changes IOO7-IOIO 

in the Tumor-bearing Host and Tumor 

Lesion 
HCl-533 Developmental Pathology and Function of 1011-1012 

the Thymus Gland and Thymic Tumors 
Wl-°33^ Incidence of Induced and Spontaneous Tumors IOI3 

in KIH Swiss Mice Used in Screening Ex- 
periments for Potential Viral Induced 

Human Neoplasms 
NCI-535 Experimental Studies on the Effect of Hu= lOl^J— 1017 

tritional Deficiencies on the Patho- 
genesis of Hepatomas in Rats 
IH;I-501 Geographic pathology field stxidies on IOI8-IOI9 

urinary bladder cancer in New QrleanS|, 

Louisiana 
HCI-50lCa) Geographic pathology field studies on 1020-1021 

uterine cancer in New York City^, Israel 

and Washii^gtonj, D. C, 
Wl'^^lih) Geographic Pathology., Experimental stiidies 1022-1023 

relating to the geographic pathology of 

mouth cancer 
NCI-52Ji- Sttidies of Polyoma Virus in Serum-Free 102i)-1026 

Tissue Culture Systems 



XIV 



!■'•-■ '-"-Pi-'- Attempt s -co ;L,?o.Late Oncogenic Viruses 1027=1029 

from Eianan Qhaaors 
NCI"532 Studies of Brain Tumors Produced in Ham=> IO30-IO32 

sters by Intracerebrally Inoculated 

Polyoaja Virus 
KGI-537 Effect of species variation on the onco- 1033 

lytic properties of Coxsackie B^ virus 

on heterotransplants of Hela cells 
NCI-538 Ttie cytopathological and cjrtocheiaical 103^ 

changes of parainfluenza vinu?. infec- 
tion in different ceil systems o 
NCI ■=539 Further observations on nucleoprotein stain- 1035 

ing with toluidine-blue molybdata 
NCI-5U0 Compailson of the oncolytic properties of IO36 

Newcastle Disease for mouse lymphoma 

cells CP388) is, viyo and in vitro 
NCI=51T Pathological Technology Section 1037-1038 

Laboratory of Physiology 

NCI-90U mtravlolet Light and Carcinogenesis IO39-IOU6 

NCI-518 Research Itevelopment Rrojects in the Field 10i^7-10i^8 

of Electronics 
ifCI-9'^0 ^Studies in Tumor<"Hoat Relationships 1049-1052 

KCT-905 Iodide Concentrating Ifechaoism of the 1053-105^ 

( Thyroid Oland 
KGI-906 Properties of Transplantable Tbyroid Tumorg 1055-1057 
UCI=910 Tumor Autoradiography IO58-IO59 

KCI-919 Investigation of Urinary Excretion of Nu- I060-IO63 

cleotides and their Congeners in Leukemia 
I»^I-920 Identification of Compound A-20 106^4- 

KC3I-925 Protein Synthesis and its Antagonism in 1065-I068 

Normal and Cancer CeHs 
K:l~927 Sialic Acid in the Thyroid Gland IO69-IO70 

WGI=911 Total Ifetabolism 1071-1075 

KCI-914 to Vivo Measurement of Tumor pH with the 1076»1077 

Capillary Glass Electrode 
NCI=915 Cellular Particulates 1078=1080 

KCI=9l6 Characterisation and Comparison of Nucleic IO8I-IO83 

AcidSj, Nucleoproteins , NucleaseSj. and 

Nuclease Inhibitors of Normal and Malig= 

nant Tissues aM of Body Fluids 
NGI=926 Purification and Characterization of the 108it-=1086 

I^oma Virus ^ansforming Agent 



X? 



Radiation 3i-anch 
KCI-688 

KCI-614.2 

KCI-659 

NC 1-671 

NCI-67^ 

HCI«676 

NCI-681 
NCI-682 
KCI-685 
NCI-686 

NCI"686Ca) 

NGI-690 

NCI-691 

NCI-69l(a) 

NCI-69l(b) 

NCI-69l(c) 
ITGI«69lCd) 



DevelopTaent^ Fabrication and Maintenance 
of Specialized Equipment for Radiation 
Research 

Antibody Production in Irradiated Mices 
Effect of Chronic and Intermittent 
Exposure to Ionizing Radiation » 

Action of Endotoxin in Irradiated Animals s 
Reduced Mortalityi Increased Resistance 
to Infection; Advanced Hematopoietic 
Recovery, 

Protective Effect of a Colchicine and Re- 
lated Coiapounds in Animals Exposed to 
X-Radiation 

Synthesis of Antibody in CoH mice bearing 
the Transplantable Plasma Cell Tumor j, 

An Investigation of the Relationship be- 
tween Cell Division and Irradiation Re- 
sponse to X-rays and Ultraviolet in Yeast. 

Applications of Some Tissue Culture Tech- 
niques to Radiation Research 

Sites of Action cf Lethal Irradiation in 
Dividing Yeast Cells 

Thymidine Uptake in Hemopoietic Tissue In- 
duced to Recover Following X-irradiation 

Recovery of Ability to Form Antibodies in 
Irradiated Mice Pretreated with Endotoxin 
or Colchicine 

Recovery Processes and Effects of Endotoxin 
and Colchicine Ti-eatments on Recovery in 
Very Ibung Animals 

The Stxidy of Basic Virus-Cell Interactions 
Using Quantitative Tissue Culture Tech" 
niques 

Repair of X-ray Damage in Cell Lines of the 
Chinese Hamster Grown in Culture 

The Dependence of Lethality on X-ray Dose 
Rate in Chinese Hamster Cells Gro^m in 
Culture 

Division Delay^ Growth Rate^ and Radiosensi- 
tivity of Chinese Hamster Cells Which 
Survive X-Irradiationo 

The extent of the Dependence of X-Ray lethal- 
ity on Chromosomal Damage in Chinese Ham- 
ster Cells Grovm in Culture o 

The Dependence or Independence of Recovery 
in Giiltured^ I-iainmaiian Cells Surviving X- 
Irradiation and Their Radiosensitivity as 
Mitigated "by Drug Action 



Pages 
1087-lOr: 

1091-1093 
109i^=■1096 

1097-1099 

1100-1101 

1102-1104 

1105-1107 
1108-111 

1111 
1112-111 

111-, 

1115-lli 

1118-1120 
1121-112? 

1123-11? 

1125-11^^ 
1127-1128 



WCI-692 A Study^of L-u... .--..-.-.ponse of Human aeU.s '-i'-> -- ' ■ 

(HeLaJ to X-Rays 
NCI-695 Studies of effects of an antibody in re- 1131-1132 

lation to development of an ascites 

(P»388} plasma cell tmior. 
mi~62k Radiation Injury and Repair 1133 

KCl-628 Dosimetry _of High Energy Radiation 113^ 

KCI-629 The Effect of Ionising Radiation on 1135-1138 

Amino Acids 
mi-Syf Development of High-Intensity X-Ray 1139 

Sovirce 
KCI-680 Radioactive Isotope Administration "by llUO-lliU 

lontqphoreses 
KCI-697 The Abstraction Reactions of Hydrogen 1142-11^^5 

Atoms in the Radiolysis of Aqueous 

Organic Solutions « 
NCI-2-650Cc] Service Radiation (Clinical Therapy) ll46 

KCI-652 Childhood Cancer 11^7 

KCI-2-65ij-Cc) Time as a Modifier of Clinical Radiation llJi8~lli^9 

Response 
NCI -2-656(0) Distribution of S-35 in Human Tissues 1150-1152 

and Tumors and the Clinical Effects of 

Large Doses of S"35= 
NCI-2-660(c) Clinical Radiation Dosimetry 1153-115^ 

NCI~2-662(c) An Appraisal of Renal Fiinction and Infec- 1155-1156 

tion in Advanced Bladder Malignancy 

Treated with Ileal Bladder Construc- 
tion and X-Irradiationo 
HCI»2-=668(c} Radioactive Isotopes in Interstitial 1157-1159 

Radiation Therapy 
HCI-2-.670(c) Studies on Thyroid Cancer II6C-II61 

KCI-693 Radiation Induced Dysplasias of Bone II62-II63 

101-694 Development of Wedge Filters for a 2 Mev Il64-ll67 

X-ray Beam 
KCI-696 Studies on Bronchogenic Caxcinociai Com- II68-II69 

parison of Combinations of Chemotherapy 

and X~Ray with X-Ray Alone 
lICI-698 "Oxygen Effect" and Tumor Size in Local II70-II71 

X^iiradiation of the C3H/BA Mammary 

Adenocarcinoma 

Surgery Branch 

NCI-10-750Cc)Removal of Cancer and Other Tissues for II72-II73 
Histologic, Biochemical and Other 
Studies as Required by Scientists and 
Other Investigators for Correlated StvidieSo 



x^ni 



NCI-10-75l(c} EvaJ-uation of Radical Surgery and. 117'+"ll82 

the Development of Mew Surgical 

Techniques as a THierapeutlc Ifeans 

of Palliative or Definitive Therapy 

of Cancer. . 

KCI"9"752(c) The Virus Treatment of Human Carcinoma II83-II96 

of the Cervix and Pyridoxine Defi- 
ciency in Conjunction with Viinas 

Treatment of Cervix Cancer- ^ 

KCI=.8"753(c) Determination of the Frequency and 1197-1206 

Significance of the Finding of 

Tumor Cells in Washings, etc. 
UCI-8"75l<. Evaluation of the Mature, Cause and 1207-1220 

Means of Preventing Seeding of Opera- 
tive Wounds Using Transplantable 

Animal Tumors. 
NCI-8-755(c) Use of a Chemically-Defined Water Solu^. 1221-1231 

ble Diet in Cancer Patients. 
KCI»9"756Cc) Virvises Associated with Human Tumors. 1232-1238 

HCI=10-759 Experimental Studies of IfechaniBms of 1239-1253 

ifetastases; etc. 
1TCI-I0".762(c) Study of ^tabolic Changes Associated 125it-1259 

with Cancer Surgery. . 

1101-10=765 Effect of Carcinogenic Agents Using a 1260-1262 

Bronchial Pouch Preparation in the 

Dog . . ^ 

HCI-10-768 The Relationship of Uropepsin Excretion 1263-1265 

to Gastric Acidity and Pepsin. 
]!iICI-=.10=.773 Study of Host Factors Associated with 1266^1272 

Implantation of Cancer Cells, etc. 

Office of the Associate Director in Charge of Research 
Summary Report 



x\n:ii 



NATIOH/i CMCm INSTITUTE 
OFFICE OF THE DIRECTOR 



Included among the responsibilities of the office are: the 
coordination of programs of the National Cancer Institute with other 
governmental and non-governmental organizations, direction of the over- 
all administration of the Institute, and the provision of administrative 
services for the Institute « 

During the past several years the increase in interest in health 
research and the miiltiplication of institutions and individuals engaged 
in research have intensified the problems of attaining the essential 
coordination and cooperationo The Dii-^ctor^ as an aid to the solution 
of these problems, seeks close ties with other organizations engaged in 
cancer research. He serves on various advisory ccamiiittees to other 
institutions and maintains constant communications to unify direction of 
the research effort. 

Important policy decisions must be made regarding research direc- 
tion, control activities and public health problems relating to cancer o 
Advisory bodies such as the National Advisory Cancer Coxincil and the 
Board of Scientific Counselors were established to aid in these deci- 
sions. These Boards are composed of outstanding leaders in fundamental 
and medical sciences relating to cancer. 

In calendar year 1959* significant advances were made in many 
areas of cancer researcho Ihese are described in other parts of the 
report o 

The increasing complexity and number of programs for vrhich the 
Research Grants Branch is responsible, the rapid expansion in volume 
of Idle number and types of research and training grants administered 
by the Branch and the increasing demands for analytical data and service 
to review boards of the Council and to administrative echelons of the 
NIH and PHB, indicated the need for revision of the organizational and 
staffing pattern of the Branch = In June, MTo Maurice Arnold was appointed 
to a new position of Operations Officer, as the first step in developing 
this expanded program. Other key staff have been added and revised 
organizational plans have been submitted to the Director, NIH. 

To accommodate the marked expansion in staffing^ particularly in 
the chemotherapy program, it was necessary to relocate the major part of 
the Biometry Branch and CCNSC and all of FIDB in the Robin Briilding, 
Silver Spring, These moves were accomplished during April, 1959 • l-Jhile we 
were reluctant to have these activities located away from Bethesda, cir- 
cumstances were such that we were compelled to operate under these condi- 
tions, l^hile the moves gave temporary relief to our immediate space needs, 
our vigorous recruilaaent program has resxilted in otir reaching the point 
where additional space must be allocated to the NCI if the prograzas are 
to go forvrard. Our space needs, which are becoming quite critical, have 
been presented to NIH management. 



-2~ 



As a result of organizational decentralization of budget manage- 
ment staff from the Financial Management Branch, to the Institutes and 
Divisions, the National Cancer Institute assumed responsibility for the 
administrative supervision, peyrolling, etc. of Mr. Fred Shaw, budget 
examiner, for the KCI, and his staff. There were fo\ir principal reasons 
for adopting this plan: 

"1. To establish clearly the budget examiners role as a 
staff assistant to line management in the Institutes 
and Divisions. 

2. To eliminate the arrangement in which some of the budget 
staff in the Institutes were financed through the central 
management fund while others were financed directly from 
the appropriations of the Institutes. 

3o To segregate clearly costs chargeable to Institute budget 
management and those chargeable to the central budget 
management function; and 

ko Facilitate Idie strengthening of the budget ex am i n er staff, 
where necessary, by relating reyuirements for such staff 
directly to the program appropriations being served rather 
than attempting to finance them throu^ the central manage- 
ment fund. 

This change was put into effect May 1st. 

As a result of considerable expansion in the research and related 
programs of the national Cancer Institute, including support of research 
in Non-Federal izistitutions in this country and abroad, there developed 
a continual increase in the number of important scientific papers sub- 
mitted to the Journal of the National Cancer Institute for publication. 
Due to Bureau of the Budget limitation on the total pagination and print- 
ing cost allowed the JNCI, there developed a significant backlog of 
scientific papers to be published. Necessary authorizations were ob- 
tained to increase the Journal to 3,000 pages (increase of 600 pages) 
and to increase the total printing costs. 

To further augment the publication needs, a monograph series was 
also initiated. 

The JNCI was honored by receiving the American Medical Wiriters 
Association award for 1959 for distinguished service in medical joiirnalism, 
Mrs, Doris Chaney, Supervisory Publications Editor of the Journal, accepted 
the award on behalf of the JNCI at the Association's l6th annual meeting 
held in St, Louis, Missouri, in October, 



There were 7^ professional people who visited the Diirector 
axid/or his scientific staff during 1959. Tt^enty were from various 
sections of the United States while the remainder were from 23 foreign 
countries » Oliis number excludes groups who either attended meetings 
or symposiums or were given a general tour of the Institute. 

A directory type report covering the Institutes Research Fellow- 
ship program for the period I938-I958 was developed and the manuscript 
sent to GPO in late 1958. The printed directory, received approximately 
July^ ^939) received extensive distribution. In addition to the regular 
PHS mailing lists, copies were sent to the libraries of all four-year 
medical schools in the U.Soj libraries of all four-year colleges in the 
U.S. (this distribution was arranged in cooperation with the Office of 
Education) j certain foreign institutions and staff members who had co- 
operated in the Research Feilovrship Program in the years represented by 
the Directory J selected list of U.Se Foundations that give grants or 
support educational work; the fellows themselves -wixo requested copies 
in response to a notice that the directory was available; other selected 
individiials to whom the Directory wotild be of interest. 

The publication of a Monthly Summaiy of the Journal of the National 
Cancer Institute was initiated in I958 to serve a nuoober of different pur- 
poses. As a means of brijaging the contents of the JoiimaJ. to an ever 
wider circle of scientists, it was decided in 1959 to contact the medical 
schools throxxghout the world. A foim letter accompanied by a copy of the 
Summary was sent to the heads of all medical schools in foreign countries 
except the few that receive the Jovimal, asking them if copies would be 
useful in their libraries. OSiere was a very gratifying respoiise to this 
offer and a number of foreign libraries and research scientists were 
added to the mailing list as a result. In the case of some important 
libraries, a complimentary subscription to the Journal was assigned to 
them. 

An Index of the Proceedings of the National Advisory Cancer Council,, 
1937-lt7 was prepared. Althougfci not in minute detail, the document covers 
kQ pages and will be of considerable help in locating various subjects 
discussed at Council meetings in the 10-year period. 

"Summaiy" histories of the work of the NCI (exclusive of details on 
researcsh programs for which source materials only have been indicated) 
have been prepared for the respective fiscal years 1939-19'+^» IJ^ some 
cases important reports made to or by the Coiincil or other documents 
have been attached to these Summaries as appendixes, \loxk. on the last 
year (19^1-6) to be covered by this project, is in progress. 



SUl-S-lAKY STAIES-JENT 

Program Activities Report, Calendar* Year I959 

Office of Information and Pub li cat lone 

Office of Director 

National Cancer Institute 



The Office of Information and Publications compriBes thiree 
Sections: Research and Program Reports^ Information and Education, 
and Reference soad DistrlTjutiono A new staff member entered on duty 
in the Research and Progreaa Reports Section to assist in meeting 
the increased demand for science writing <> A six months' effort to 
recruit a Publications Writer (GS"ll} to replace a staff member who 
was promoted and transferred to the Information and Education Section 
was still unsuccessful by the end of the yeay, 

Tae Information Officer was appointed by the Director as an 
HCI representative on the Planning Caamittee for the Fourth Hational 
Cancer Ccaifereace t-o be held in Minneapolis September 13-15, I96O0 
In tMs capacity he attended several meetings of the Committee aad^ 
as a member of a subcommittee on esiiiibits, ccaif erred in Minneapolis 
with representatives of the University of i-linnesota, the Mayo 
Foundation^ ^ad Mirsnesota State Health Iteparlaient on exfaibltg end 
related subjects ^ 

Sie Erebiozea controversy occupied a good deal of the 
Information Officer's attention during the year.. Thi® included 
response to a number of press and Congressional inquiries and the 
preparation of several explanatory documents , He prepared a state- 
ment for the Director in response to a new Krebiozen test approach 
suggested by the Chicago DsJ.ly News, a position statement for ttie 
Surgeon General to use if asked for comment at a press conference, a 
background statement for the Secretsa^y's use in a press conference, 
and a chronology of eve»t3 involving the Government in the Krebiozen 
controversy !lfee last was done at the request of the House Committee 
on Interstate and Foreign ConHEerce. 

A special report giving the position of the Public Heeli^ 
Service on smoking and health, and summarizing the findings of 
laboratory end biost^tistieal investigations on cigarette smoking 
and lung c£sncer, was prepared by the Infosssatioa Officer for the 
appropriation hearings as requested by the House subcommittee » By 
invitation from the American Cai^cer Society, the Information Officer 
attended meetiags in Boston of the Society's Ad Hoc Committee on 
Smoking ©ad Health, and its Information end Education Committee, to 
hear policy discussions on smoking and lung cesscero 

'fhe Information Officer was appointed as a member of an MIH 
Task Force to review the civil service classification standards for 
information positions, attended several meetings of the committee and 
participated in a study of existing and proposed new standardiSo He 



- 2 



also served with the Infonnation Officer, BSS, and the Infonaatlon 
Office?^ B53, on a paael to screen eligible GS-12 iafonnatioia 
specialists in PSS for a new GS»13 Information officer positioa 
in the Caacer Costrol Progrsm, ESS. 

At the request of the Director > the Information Offieer 
consulted with Deputy Canmissiaaer J.. Lo Harvey and Mo Ro Stepfeeafi, 
Director of the Bureau of Enforcemeat, FDA, concerning proposed 
California legislation to control caacer quackeryc On the basis of 
this discussion he prepared a letter to State Senator Richard Richards, 
chairaiaa of the coBimittee handling the bill, giving the Institute's 
and HL4 views on the subjecto 

ITae Information Officer took the leadership in planning the 
Institute's second Cancer Information CcHiference for high school 
science teachers of the Washington area and nearby region, which wae 
held on October 17= The program was worked out in collaboration with 
the national Science Teachers Association and the Uo So Office of 
Education o In the spring he arranged a special pro-am for, and 
gave the welcoiaing address to a @x3up of high school science teachers 
frcm Roanoke 5, Virginia, who made a one-day visit to HCIo 

Mauy of the ccmtacts with the New Ifork and Washington offices 
of Time Magazine, resulting in a cover-story on HCI and cancer research 
in the July 27 issue ^ were handled by the Infonaation Officer beginniag 
in Febnierjr when the magazine made its first investigation of picture 
possibilities o He also participated in conferences over a period of 
several months with representatives of the British Broadca«tlng 
Corporation leading up to the filming of a cancer research story at 
KCI for two televlsi<»^ programs to be broadcast in Europe and later 
in the toited States. 

In collaboratioa with the Chief, Biometry Branch, the Information 
Officer developed deslg^is and ccaxtent for several ejdiiblts on inter- 
national cancer programs and the world-wide distribution of cancer as 
requested by the Senate Conaiittee on Qovemment Operations for hearings 
on the Intematioaal Health Institute bill. 

The Information Officer was occupied during the year witti 
probleias related to the digsoaiaation of scientific information throu^ 
publications and reports » He prepared the 'documents required for 
Bepartmeatal request to the Bureau of the Budget to amend the Bureau's 
authorization for publication of the Jour&al of the National Cancer 
Institute as a periodical, to allow an increase in total pagination 
frc»n 2,it00 to 3,000 a ye^-, and a larger printing budget to cover in- 
creased costs and pejsnit the printing of additicaaial color plates » Ee 
also participated in arrangements for clearance and printing of Caacer 
Chemotherapy Reports as a regular series of administrative reports o 



He also suggested to the Director, in the interest of feeding up 
coimamication of scientific information^ th&t autfeors' Bunrnjaxies 
of articles distributed with publication of each issue of the Journal 
of the National Cancer Institute "be reorganized to list monthly, 
suiumaries of all papers accepted for publication in the JNClg Cancer, 
and Csacer Research, with the date of publication indicated where 
possible o 

^8 Information Officer participated in a conference with the 
Director, NCE; the Chief, Cancer Control Breach, and information Officer, 
BSSj to discuss the relative roles of Nil and BSS in cancer control 
progranss = 

During the year OIP handled l82 inquiries froa newspapers, 
new services, free lance writers, radio and television stations o 

A total of 7^ itemfi were processed through cleasrance, includii^g 
68 speeches, articles, press releases, aipecial statements and other 
materialg produced by the information staff. !HEe remainder were papers 
submitted for clearance by other Institute staff members » 

Reaeaycfe aad Pro-am Reportg Sectlm — Materials for use in 
the aeas-ingg on NCI appropriations for the ST i960 Budget included s 
conrpletiffii of report, "Higlili^ts of Program Developments, 1959 j" 
report on status of tlje program of the Cancer Chemotherapy National 
Service Center, 19 58 I report on the scope of NCI graat-supported 
resesych in support of the Secretary's revest to the Aaministratioa 
for additional ftusds in the Budget; aisd insert for inclusion in the 
House subciMjmlttee report o Materials prepared for the Searings on 
the F£ 1961 Budget included: Director's opening statement and report 
on highli^ts of program developments and research studies conducted 
and supported by NCx, 1959 « 

Materials prepared in response to special Congressional 
requests included.; statement on caacer research and the prospects 
for iiie control of cancer for Senator Dirksen, a similar statement 
for Senator Msindt, and & background statement on the chemotherapy 
program for the Committee for Social Legislation c 

Ibe weekly report to the Director, NIK, contained a total of 
75 items, including 22 on program highlights and 53 on research 
highlights summarizing papers ftrom 22 journals » Joumsils most f^i^ently 
cited were the Journal of the Katioaal Cancer Institute easd Proceeding a 
of •ate Society for Experimental Biology sad Medicine o 

Seven susmisries of scientific papers presented at meetinigs of 
the Merican AssociaticaE for Cancer Research, Federation of American 
Societies for Escperimental Biology, and American Chemical Society 
were preparedu Two press releases on new chemotherapy contracts were 
also iasuedo Assistance was given to about 15 science writers who 
visited the office =. They received source material on cextcer research. 



.. ll « 



statements oa cancer reseexcli for attrilautioQ. to the Director, and 
appointments with NCI scientists and officials o In connection with 
the cfeemotherapy clinical aymposium held in Washington in Hovember, 
a press release announcing the meeting v/as prepared for science 
writers and a press room was organized and staffed., 

Eight review papers for presentatian and subsequent publicatioi*, 
were prepared for the Director o A "staff document" on the chemotherapy 
patent policy was prepared at the request of the director of the 
chemotherapy program and a brief statement on leukemia research waa 
prepared for his use in a radio broadcasts AXso^ for his use at the 
American Cancer Society's meeting for science writers at Excelsior 
Springs, a revised backgroxmd statement on the chemotherapy program 
wajs prepared^ At the reqoaest of the Assistant Director, a form 
stateraent on the treatment of cancer and the present clinical use of 
chemotherapeutic agents was preparedo 

Miscellaneous other materials that were prepared dmring the 
year included the NCI portion of the I959 anniial report of the 
Secretary, WE^; speech materials on cancer for use by the Secretary 
at the conference -symposium of the Eleanor Roosevelt Institute for 
Cancer Research and the AFL«CIO in New York; a lay informatlcaa 
enclosure on cancer research today for use in answering inquiries 
from the general public; a list of qpiestions and answers on cancer 
chosiotherapy for inclusion in an American exfeibit scheduled for 
showing in Ifea cow; and a 10-«iiiaite quest ion -and-aasawer script on 
cancer research for a staff scientists who was interviewed in a 
radio broadcast o 

Staff members represented the Office of Information and 
Publications at the meeting of the American Association for Cancer 
Research in Atlantic City, the First Annual PBS Information Officers » 
Workshop in Atlanta, "Uze Second Science Demoestraticaas Ccmference 
for High School Science Teachers in Bethesda, and two meetings of 
the Cancer Cehmotherapy Sational Ccasnltteeo 

Information and E(3ueatien Section — Ei^t articles were 
written for byline of the Director » Six of these were for lay or 
professional ;}oumals, including XAfe and lealth, the Science Teacher j 
Public Health Reports, Geriatrics, and Modem Medicine; one for the 
American Peoples Encyclopedia Yearbook, and one for the Robert So Allen 
syndicated newspaper column » Releases for the press totalled nine^ 
Two TV scripts were written for gixest appearances by the Director on 
local pro-am® » 

Two speeches were prepared for the Direct-or« In caaes where 
complete drafts were not required for his speaking engagements, the 
Section provides him with several copies of "Remarks," essentially 
a summary of a scheduled talk, which he can hand out to the press 
if asked for quot^le materialo Advance copies of these summaries 
are often requested by the host organizaticm's public relations 
representative for publicity purposes » Hine such "Remarks" were prepared » 



5 - 



speech materials vere al^o prepared for •th.Q Surgeon GeaereQ. 
on one occasion o 

Prociuotion of a speech outline and sketches of accompany- 
ing illustrations for slides vas caapletedo This OIP project is 
for ane of the Director or any staff meniber who desires to use it 
in maJjing talk® 'before lay, civic, or aub-profesaional groups on 
csncer research or eontrolo The slides are keyed to the outline, 
any parts of which may "be used, according to the interest and 
level of the aadience.. Working title for the project is "Let's 
Talk About Cancer =" 

A special report on the cancer diagnostic test program was 
prepared for use in the Hearings on NCI appropriations for P^ 1960,> 

The KCI "brochure was updated and releasedo This was the 
second revision since the publication was originally released in 
1955'^ Also revised was "Reading on Cancer-" Both of these 
publications are widely used in response to public inquiries o 
A new pamphlet, "Bibliography on Cancer for Nurses," was cleared, 
and processed for printing in the Section c "Highli^ts of Process 
in Research on Cancer, 1958" was also publishedo 

Inquiries from the public and Congressiaial sources continued 
to increase in volume, a trend which began in l957o In order to 
keep pace with this load, a nisDfl>er of the Section staff was assi^ed 
full "time to assist the public ia^iries information specialist o 
Letter replies to public inquiries totalled 1,517, representing a 
37 per ceat increase over I9580 Of the total, 184 inquiries were 
frcan CoB©°esai<mal scsirceso 13ais vs& less than were received in 
1957# chie "to a decrea5:e in inquiries on Krebiozoio Still, 101 
Congressional letter* were concerned with Krebiozeno Over 2^600 
pieces of informational literature were distributed with replies » 
Telephone inquiriea to tfee public inquiries unit totalled 638, or 
an average of 12 » 2 per week., 

Activities in the visual aids area included the selection of 
appropriate photos and captions in reply to reporters" requests to 
accoBipaay rfccry material, or to illustrate articles prepared in OS*; 
planning and rough design of charts or other visual aids, including 
a special set of charts on the international aspects of cancer for 
Senator Husipbyey and drafiis of 6 charts for use by a Senate Committee; 
and supervision and scheduling of ejdiibitSo "Cancer, Where We Stand 
Today^" a popular lay e:daibit, was revised and redesignedo The visual 
inforraaticaa specialist also wrote articles for magazines, speeches 
for the Director |. summaries or "Remarks" of his talks, and other 
miscellaneous writing assignments whether or not they included visual 
aid materials o 

A member of the Section was assigned as NCI reporter for the 
newj enlarged FIH Be cord ■> 



Reference aod DistrlTautlon Section — The scTeenlng of 
selected journals far papers published by KCI staff personnel or 
grantees, for use by the vriters of the Section, continued.. During 
the year an average of 88 journals were screened monthly, tjcom. 
vhich a total of 1,569 reference cards vere made on papeirs reporting 
cancer -related findings » 

Bulk distribution of lay and professional publications and 
reprints on various aspects of cancer research and related activities 
totalled 1^1,232 D This represented a 62 per cent increase over 19 580 

SadlvidMal requests which come from students, teachers, 
professional and service organizations and the general public, also 
increased during the yearo These requests, which do not generally 
require letter replies, totalled 2,ll6 or an increase of 6I per cent 
over 1957« to response, 9^758 pieces of printed material were sento 

Distribution of press releases and announcements to science 
writers and professional journals totalled 3*575* and monthly 
eummaries of papers appearing in the Journal of Idle National Cancer 
l32Stitute, which are mailed to science writers and professional 
joumeJLs, 8,400o Film bookings for diagnostic and lay films 
numbered 112, with 72 referrals to the regional offices for 
"Challenges Science Against Cancer o" 



Program Activities Report 

Calendar Year 1959 
joximal of tlie National Cancer Institute 



The Board of Editors reviewed 6o4 items for piiblication this 
year. Of this nimiber, 22i»- were manuscripts tj WCI staff for publi- 
cation in other scientific or medical jci'imals--220 were approved, 
3 were rejected, 1 is pending! 23^ (57 t>y NCI staff and 177 "by 
authors outside) were maauscriptG subraltted for publication in JlfCI 
—165 were approved^ 34 were rejected, 2 were withdra-sm, and 33 are 
pending o OSisre were Ikh abstracts and book reviews approved and 
2 are pending o 



Volvimes 22 and 23 contained 2,696 printed pages « Issues of 
250 pages are scheduled thro\i@a May I96O and 68O printed pages are 
being reviewed for JNCIo 



An nCI Monograpii series was established tMs year= lixe first 
monograpih will be published in December 1959 an^d 3 additional mono- 
graphs are being prepared for publication in I96O0 



She monthly suramary service established last year has been 
popular and HSL institutions and individuals are receiving the sucm;: 
rieso 



The American Medical Writers ' Association presented the Jotii-Tial 
with its 1959 honor award for distinguished service in medical 
journalism (class 2— medical specialties ) o 



10 



ary Report . 1954>1959 



In the fsll of 1954, eh@ p¥@@@tit Ghlfif of th® Biometry 
IrsncSs prepared a ragmossndum for the Dlt'ector o£ $h« NaeionsI 
Cancer Institute , @atltl@il, "A research program in blem&tsy @ad 
epidtsmlology of cancer o" Review of thia docuissnt iadica£e@ thst 
eh@ g@n@r3l objectives snd sppirosches hsv€ been tzisintained during 
tha fiv@ years that have now elapsed „ 

Biometry, it is bell«v@d, hss gained new statur@ and has 
contributed ies ahare to the advances in csnccr research, over 
sm £r@a much wider than the Biometry Brsnch itself. 

Biomgtry has an important rol@ in several other progrsra§ 
of the ISstional Cancer Institute: (l) It assigned key persmutel 
and continues to develop th<t staff and istatistical program of 
the Cancer Chemotherapy Hatlonal S<grvio« Center^ (2} It 
initiated and directs the end results ^ogram as s collaborative 
ventur® with the CCNSC; (3) It has assigned k©y personnel mtd 
continues to cooperate in the statistical needs of the Field 
Investigations and Development Branch; (4) It collaborates and 
assists in projects in geographic pathology undertaken by the 
Laboratory of Pathology; (5) It collabcifatgs and assists it. 
statistical problems of various laboratory studies, paffiicularly 
in chemotherapy. The basic philosophy ©f the role of biometry 
is to consider each need on its own merits: Botm needs can be 
met best by retaining the technical function in the branch; 
oth@r naeds require the creation of district units in other 
Branches, retaining consultative relations onlyo 

Biometry consults and cooperates in various capacities 
with the Cancer Control Branch of the Bureau of State Services, 
with the National Office of Vital Statistics, and with non- 
government agencies such as the An^ricnn Cancer Society and the 
Sloan-Kettering Institute. Such collaboration is of great 
asaistanc© in the progress of ©ur own Braaich programs and we 
hope, to the other organizations concerned o The Branch is 
increasingly represented in national aad international consulta- 
tive bodies and meetings concerned with bioai@try and 
epidemiology of cancer o 

The research contributions of ths Bicva^cry Branch during 
the past flvm years are recorded in over 75 separate publications 
that have appeared with various msmb@rs of its staff as authors „ 



1-1 
\ 



- 2 - 



Among t:he vm&t itepext&nt isoQtributioQe ase the svsrveys 
and asalyees of igaBecs> iesldsases mortality and sunriTal in tha 
United Statass "Morbidity fsom Oamge? in the IMited States" 
CPS Menegraph 56 » 1959)9 "eaae« Herbidity in Ikbaa and Eusal 
le^^a" (PS MenoggeAph 37, 1956), aad "Caages in Cenneetieut 1935- 
1951." Thie basi©;, d«tall«d infemation re€0?ds a lailes&ons 
in th« epldemielo^ @f gan^aro It was avEsussixisd in a pepula'f 
pi&l£eati©np "Th« Eseent ©f Caager Illness in th« United Seatss," 
CPl Publo 547^ 1958), &t^ modified into the influential Senate 
psint, "Cactger- a World-Wide Manage" (1959) o 

Siecaet^ Branch has been the fe<§al point ef studies and 
analyses &S the role ®£ emciiing add lung ©snger^ iihish <suliaiiia- 
t&d in th@ @ffi<gi8l Public Health Senri@e etateacnts <m this 
s»&j@@e ®a July 12^ 1957 and on Kovember 28^, 1959 (JAMA 171? 
1329) o The 3san@h ©oadugted the first aaalyvis of t«3ba(g«s© 
@OBSteapti©n in the Ifnited States CPH Msaograph 45, 1956), 
©asried eut a national study on @ait@e¥ ef the l«ing in woraen 
iMlQl 21? 825s, 1958) e and is ©ondueting a ea^i©aal analysis 
of ts^^upatlonj, £'eslden@e and siaeklngo We havs pasti@ipa£@d 
in national study ^oups (Seiei^ej, I25s 1129^ 1957) and reviews 
©a the tople iJmi 22| 173 ^ 1959) » 

Th® aatioaal end-results psegra® {&£ 3mi 22s 633, 1959) 
is beginning to genei^ate reliable^ representative info¥taaf:ioQ 
ipegfirding ©anser patisntSj, %«hat happens to thgtn^ liliat'eficests 
various treataents have on theig* disease^ and ^at fa@to?e 
laodify their ©linisal @ours@o This is th@ natural development 
of ous* Icng-texm @oamitiaeats in Connegtisuts) and oue pioneer 
psesentation of a p^ograia on togal end-results in (gangtes* at 
the Thisd Mati@nal Caneer Conference C^o^eediffigSg 1957),, lAigh 
is nmr being esctended on an international S€al@o At the sante 
tiioe, fflors detailed aaaljesa "in depth" are on the way gnu ©an'ser 
of the <getvis£ a&d body of th« uterus^ on breast (JISOI 23s 1137^, 
1959)8 and other spc(si£i@ sites. 

As isap^rtant as any Bpe@ifi<g eontrlbution has been the 
influ®s@e eietending t© @liniisal and laboz'atosj investigateofs who 
are in@reasingly using es^erinental desigp and etatistitsal 
analysis in tb&is mm etudieso to stiieulate this developmgatj, 
a nissber of papers on laethodology have been p^epared^ su@h as 
&R the rets-@@pe@tiva stethods of study in disease (JIOZ lis 1269 » 
1951 I 22s 719, 1959 5 23s 487, 1959) o These papers have been 
well X'e@eiv@d and widely tssedo 



12 



3 « 



Tlie grcwth of the importance of the Biometiry Branch is 
IntimAtely aeso^lAted with the continued trainiag and growth in 
professional stature of its senior investigators. The selestion 
of Mr. Marvin Schneidenaan as a Ro<skefeIler Research Fellow for 
a year's study in England to complete his Ph.D. eourse, and the 
assignment of Itr. Sidney Cutler to the University of Pittsburgh 
Sehool of Public Health to c(»nplete his requircioents for a 
doetorate are evidenses of thii> growth. The Bramsh initially 
developed a training arrangement at Plttsbi^gh, in realization 
,o£ Its future and eontinulng needs, which is now a HIH program 
involving all schools of public health. The Branch has had 
several exeellent sutoner students, some of whom will join our 
activities eventually. A number of Visiting Scientists from 
England have been a refreshing sour@e of new contacts, some of 
whleh are developing into collaborative studies. 

During the past year the Branch has @ontinued its 
general program of activities, with the same personnel and 
approximately the same budget that it has had for the past 
several years. Two developments are singled out for notes 

(1) Internatlginal program . A pro^nlsingg euthusiastie 
sollaborative program in end-results and epidemiology of @an&er 
Is now under way between the United States^ Detsoark, England ^ 
Finland, Franse and Norway. Coordination of definitions and 
criteria Is being achieved for the purpose of presenting a pro- 
gram on ean@er end-results at the Eighth International Cancer 
Congress in 1962. Arrangements have been inade for a pathology 
review of cervical (gainer in Conneetleut and in England^, to 
examine dlssrepancies in the distribution and stirvlval in this 
neoplastic entity between the two geographic regions. 

(2) Intensive use of mortality data . Collaboration with 
the National Offlse of Vital Statistics has permitted mo^e 
intensive use of mortality data for epidemiologic studies. 
Analyses of regional variations and of mortality aiziong native 
and foreign-born individuals are ncaring eompletlon, and reveal 
some Interesting variations and trends. Analysis of residence, 
otesupatlon and smoking habits of a lO'perisenC sample of the 
1958 lung cancer deaths will add new correlated information to 
this important field o The excellent response to this type of 
study indicates that a similar design should be considered for 
wider research application. 

What of the future? A forecast pr&pared on November 21, 
1958;, in a memorandum to the Executive Officer of the National 
Cancer Institute, recommends an expanded program in biometry and 
epidemiology, and a ©loser reintegration of these aetivltlee which 
were administratively separated in 1956 <. 



13 



I 



Visualised in tuis forecast are at least two "research 
centers ' in Connecticut and in California, b^sed upon popula- 
tion laboratories that can be developed there. ^Iso suggested 
was further e3q>i&n8ioa of the international prograia, with the 
anstive conjunction of pathology and epidemiology. ''An imagina- 
tive program in the epidemiology of cancer, not limited to 
statistics but including patiiology and carcinogenesis labora° , 
toi?ies in the United States Jind other eounts'ies o <> » o o would 
require between 1.2 and 1.3 milliono .» » . . <>'*. 



Ill 



i, iiioutetry Branch 
2o Office of the Chief 
PHS-NIH 3, Silver Spring, Mdo 

Individual Project Report 
Calendar Year 1959 

Part A» 

Project Title: Activities during 1959 

Principal Investigator: Mo B= Shitakin 

Other Investigators: Hone 

Cooperating Units: Cancer Chemotherapy National Service Center 
Journal of the national Cancer Institute 
Division of International Health 

Man Years (calendar year 1959) : 
Total: 3 
Professional : 1 
Other : 2 

Project Description: The activities of the Office of the Chief have 
included the follotd.ng: 

(1) General planning and direction of the scientific progipans 

of the Branch, with the collaboration of the Associate Chief, 
Mr, William Hasnszal, including developstent of international 
cooperation of tumor registries » 

(2) General supervision over clinical studies of the Cancer 
Chemotherapy National Service Center, as Assistant Chief for 
Clinical Studies » 

(3) Scientific editorship of the Journal of the Kational Cancer 
Institute, including initiation of nera series of monog^-aphs o 

(4) Liaison and NCI participation in development of plans for 
Hatioaal Institufee for International Medical Research, and 
in various phases of USA-USSR msdlcal relationships as 
requested by PHSo 

(5) Participation in preparation of prints for Uo S„ Senate 
Conmiittee on Government Operations {Cancer: A World-Wide 
Menace, July 17, 1959) » 

(6) Investigations of certain historical aspects of cancer 
research and of induction and growth rate of tussors, 

(7) Consultation on cancer epidemiology and cancer control to 
Cancer Control Branch (BSS), and American Cancer Society, 

Part B included: Yes 

.. 15 



Seffiai lo. NCss50_ 

ri.:. S? Honors, kjjatxls ^ and Ptsblieafcioag 

I-^abiicatioR8 othsr than abstracts froa this projects 

Shiikla, M B.': Jacob Wolff, Historian and Biographer of Ceacer. 
Cancer 12: i-iii, 1959 

C43rQfield, J.; UaBnsz&l, M.; Ha^aoad, S. C; Lilieafeld, A. M.; 
Shiskin, M. B. sad fades', E. L. s Seaoking aad Lung Caacers 
Kaw Evideacs as^d a Discussion of So®© Questions. J. Nat. 

Cancer last. 22s 173-203, 1959 

Slilialtiaj, M.- B.s Katitral History of neoplastic Disease. In 

Fbysiogathology of Gaacer <ed. F. loaburges) ed. 2. pp. 855- 
871. Kaw Yo^k, 1959. Hoaber -Harper. 

Shimkin, M. B. aad Fields ^- B; Ch^iotherspy of Caneaie, Ie 

C&nsBX. gjagnosig and Yreatmeat i&d. J. B. Field) pp. 724- 
751. BostOQ, 1953. Brown & Co. 

ShiEkia, M. B. aad MacLeod, C. M. s Medical Education In the SFSSS. 
jr. Mad. Sducafcloss 34; 795-801, 1959. 

Haeassel, ^. and Shiaskia, M. B.s Ijuzag Cancer Among Women. Acta 
Bhio Isteraat. Contra Gaacsusa 15 2 493-501, 1959. 

Shlfakiaij M. B.; On the Etiology of Broachogenic Cas-cincsaa. In 

Biaffliosis aad Tfea tmssat o f Tumors of the Chegt (ed. D. Spaia). 
Gruea aad Steatton. In press. 

Shi^in, M. B.s The Spid^iology of Cancer. Modes-n Medicine. 

In press. 

ShJiskln, M. B. s Science GosKsunication Sox^ the Twentieth Century; 
A Daily Hewspaper of Bicasedicai Research. Mississippi Vallsy 
Med. J. la press. 

Sht^iBj M. B.J ^ehiir Hanau: A Pusrthar ISote on the History of 

TiSKor Transplantation. Cancer. In press. 

Andervoats H. B. and Shimkia, M. B. j festieulas TimotB in Mies 
af ts^ ;. Rsao'^'al of Seilbestrol-cholestexol Pallets. Aeta enio 
lateraat. Coats-s Canesum. In press. 

MdeTvont, E. B.; Shisakinj M, B. and Gsntg^a H. Y. ; The GrovJth of 
Estrogen-iadissed Intes'stitial Call Tusaosrs in BALB/c Miee. 
J. Mat. Cancer Ins.t. In p?3ss. 

HoRoss and Awayds relating to thiss project; 

Jou'^aal of th® National Cancer Institute was the American Ifedieine 
Wifites-'e Association honoff award for distinguished segviee 
in afedieal journal is® (Oc.t„ 1959). 



16 



2. I?«8ogse^,hy Ssctioa 
pjjg.jjjH 3. Silver Spffing, Md. 

Individual Fsfojecs; Repox^t 
ealeadsff Yeas 1959 

l^oj©st Tiel@? Gs&emw Morbidity S«srv®ys ia the ii»it«d States 

Prinelfial Investigator: milisia mmoM&®l 

Other Investigators ; Donald loveland and X^onatd Ohlfisse 

Goopeiratlng Holts s Hone 

iteti Tsafs Ceal«nd«ff ysar 1959) 
fofcal; 1/2 

Frofessional; 1/2 
©Khs^s - 

Ps'ajeet Be$@siftlons 

(A) Oblectivesi; To provide ba®i® data on eaa€:ejf incidence, 

pwmvalmncm, laoetality, >and survival espasrisnee in the I'aitsd 
States, Investigate @f£ect of v&sio%m detsographi® faetorSj 
Stteh a® saee^ ses, ag® gsographic region and eeonoaie status. 

CB| Methods empjLoysd ; Infosiaation on eas^®? patienta '«®®n duriag 
.& sf^elfied ealendar year is ebtain®d froa vistually all 
diagnot.ti® ®o«r®@s (hoipitaij eliaicj labos-atogy, ^hysieians) 
in ea€h area su!rvey®d. Li®tg of diagnossd cases mxe subsaquently 
%@arch@d agaieist death indexes in vital statistics offices. 
Hospitals and physieiana are also eontected for ftsrthes' 
information on p.stient status. 

' (C) Major fioding ^; So»e of the earlier fiadiegs on eancar 
iaetdenca frosa the ten-elty saofbidity surveys have b®@n 
ptablished iu Fufalic Health Monogsrafjh !fc.29. Morbidity ftom 
Eaiaeag in the Paited States - The second monogsaph in this 
®®ifi«® ptabliehed at the and of 1958 „ discussed intercity varia- 
tion in eaaeeir incidence, irslationship of eaaeer incideaee to 
economic statu®, changes in eaacer aorbidity batwssn 1937 and 
lli47, sg® distribution of cases diagnosed at different stages 
81^ a review of the eonsistency of eaaeer ineidenea data f?<^ 
vafiossg souscs-B. gsnggg Mogbidity ia Urban and Rugal Iow a 
appeared as Fublie Health Monograph Ko, 37 » 



17 



Part A - continued Serial Ho. NOT 52 



la three cities (Ballaa, Chicago, Pittsburgh) and the State of 
lova, the diagtioeed cases enusierstsd in the inorbltUty surveys 
have been traced to obtaic survival information for a five- 
year period o Searchee against death indexes in vital 
statistics offices have been consplated in the four areas , and 
this data has been coded and punched for use in XSM tabulations o 

CD) Significance to the pro gram of the Institute ; Provides basic 
epidesnio logical information regarding the incidence of cancer o 

The survival experience of cancer patients enumerated in 
saorbidity surveys will prove useful in resolving questions 
raised about the coniparability of data secured by case 
register and morbidity survey techniques .> 

(E) Proposed course of project ; Additional analyses of the ten- 
city incidence data are continuing » Special attention io 
being given to geographic variation within the cities for 
patterns of incidence . that might provide epidemiological leads » 

A monograph will ba prepared analyzing the five-year survival 
rates for diagnosed cases in the morbidity surveys and relatiisg 
the findings to other published experience, Work on this later 
phase is being del&yed until the IBM 650 computer program for 
transmitting survivsl rates can be adapted for this study » 
The conaputing program will b4 ready for tabulations in 1960, 

Part B included: Yes 



18 



I 

1 



Serial lo.NOI;^;f _ 
Farfc 3: Konoi's, Awards, &nd Publicstioas 

f«blications othe? than abstracts frosj this project: 

Do&n, H. F. and Cutler, S„ J„: Morbidity from GsKcer in the 
United States » 'Basho D. C,> U.. S. G©v= Priat. Off, Public 
Health Monograph Ko, 56. 3.959 » 

Honors and A%?ards relating to this project: 



19 



December Serial No. NCI" 53 

1. Biometry Branch 

2. DfflBOgraphy Section 

3. Silver Spring, m^ 
PHS-NIH 

Individual Project Report 
Calendar Year 1959 

Part A. 

Project title: Statistical Studies of Cancer in Pittsburgh Area 

Principal Investigators s Nilliaia Haenssel and Antonio Ciocco 

Other Imrestigatorss Dorothy Perrott 

Co®pera£ing Units s University of Pittsburgh School of Public Health 

Man Years (calendar year)? 
Totals 2% 
Professionals 1% 
Others 1 

Project Bescriptlons 

W 0b;jecttv«8 S To pursise a series of intensive cancer morbidity 
studies in a single area, to yield Information on factors con<» 
tribating to unusual incidence for specified sites. The 
collation of data obtained by a variety of approaches to the 
same population base will peztait stronger inferences to he 
drava froca the chain of evidence, open the way to designing 
detailed studies to test specific hypotheses suggested by the 
evidence, and sake possible methodological contribatieas to 
field studies of chronic illness is. human populations. 

<B) Methods employed s In cooperation with the Pennsylvania State 
Health Department aad Pitt8b».firgh City Health Departments a 
cancer register has been established. The State Health 
Department has designated the School of Public Health as its 
agent for receiving cancer reports from hospitals in Allegheny 
County. The reports froia hospitals are being supplesaented by 
a caavass of records of physiciaes in private practice started 
in June 1959, covering patients diagnosed and treated in 1957 
and 1958? the si^pleraent.ary field work will be coiapleted in 
1960. Analysis of cancer incidence by site for small areas 
%?lthin Allegheiry County will start on cosapletion of the field 



Plans for the succeeding phases of the Pittsburgh studies 
remain unchanged. The analyses of morbidity data by census 
tracts will s?^ply leads for the design of studies to test 



20 



Part A - continued Sesrlal No. MCI-^3 



specific hypotheses, ^^Ich would open the second phase of 
the project. Such studies would utilize the unique sotjrce 
of itifonoation available locally, such as: the "population 
laboratory" in the Airsenal Health Msfcrict and 1% city 
8awpl&, and the resources of the Department of Occupation 
Health (in the School of Public Health) in gaining access 
to records ©f people working in plants in the Pittsburgh 
area. 

(C) Maijor findings; By the beginning of 1958 the system of 
obtaining reports on newly-diagnosed cancer patients on 

a current basis was extended to cover all Allegheny County. 
The collected material is being entered on punch cards 
which will permit analysis of intra-area variation in 
cancer incidence for specific sites within Pittsburgh and 
Allegheny Covmty. 

(D) Sigaif leasee to the prog ram of the Ins titute ; The basic 
notion of epidemiological studies of a population in depth 
has never been attempted on this scale for cancer. As 
mentioned in the objectives, these studies will conts'lbute 
valuable Information on methodology in field studies of 
chronic illness in human populations. 

(E) Pro posed course of project : See A. 



Part B included^ 



21 



Serial No.Mfi|54, 



lo Biometry Branch 
2. Donography Sectioo 
PHS-MIH 3. Silver Spring, Mdo 

Individual Projaet Report 
Calendar Year 1959 

Part Ao 

Project Title; Studies on Caneer in Connecticut 

Principal laveetigators; Sidney J. Cutler aiKi William Baenexel 

Other Investigators: Jolia Bailar 

Has Tears (calendar year 1959); 
Total: 3°l/2 
Professional: 2-1/2 
Others 1 

Projeist Descriptions 

(A) Objeetives s Provide data on cai^cr incidence, .oethods of 

treatment, and end results. Investigate the effect on incidence 
of epidemiologic factors su@h as iirban envlronoent, nativity, 
economic status, ocst^ation, diet, and personal habits. Study 
in detail the inter-relatianship between the condition of the 
patient at diagnosis ^ the therapeutic course, and the end 
results. 

CB) Methods Employed ; Eospitals accounting for 9451 of the approved 
general hospital beds in the state participate in a ease 
reporting and follow-up program adaaiaistered by the State 
Health Departioent. 

Samples of cases reported to Che register are also dravn for 
special study involving the esi^nination of hospital records 
for additional ©linieal details and the isollection of histories 
from patients and relatives. 

(C) Ma lor findings : A con^rehenaive statistical review ©f the 

esperience in Connecticut during the period 1935-1951 has been 
reported in a laonog^aph and a series of related papers, l^ith 
the addition of more resent infonaationi, the 1935-51 data 
frcm Conneetieut have continued to be the basis for planning 
the analyses for all registers parti@ipating in the £nd llesults 
Program and the means by which new teehnisal procedures @an 
be evaluated. 



22 



pgvt A - continued Serial HOoNCI 54 



During ths past yesr several papers have been completed 
concerning the long-term survival experience of patients with 
cancer of the female reproductive aysteoio Soiae of the 
findings are: 

(a) Patients with cancer of the breast, cervix, and 
corpus who were diagnosed during the period 1945-54 had a markedly 
higher survival rate than those diagnosed during 1935-44, 

Patients with breast cancer showed no improvement in 
lO-yesir survival despite isiproveinent in the proportion 
surviving 5 years- 

Cb) Patients ^o survived 5 ygars had thereafter a 
mortality rate still far in excess of that expected on the 
basis of the general populsition experience o 

(c) Surgery was strikingly successful in treating women 
under age 55 with localized cancer of the cerviSo 

The iBsthodologlcal advances Included the development of 
a simplified, approximate method for coinputing the standard 
errors of survival rsteso 

The editing and coding of detailed clinical data in £he 
re-examination of Connecticut hospital rscorda for uterine 
cancer cases has been completed o Punch cmrds have been prepared 
and tabulations h^ve been programmed for the IBM 650 computeir 

at ^m. 

The pilot study concerning the familisl distribution 
of cancer of selected sites in the State of Connecticut has 
been completed. The study developed the methodology and 
practicable plans for a larger investigation now being under- 
taken by the. Connecticut State Department of Health supported 
by a Research Grant, 

(ID) Significance to the prograta of the Institute ; Provides basic 
' epidemiologic information regarding the incidence of cancer 
and data for evaluating therapy in terms of end results. Also 
provides mssns of estimating costs snd difficulties of 
conducting analyses on the larger series of dsta of the End 
Results Progrsim, 



23 



Parfc A -• continued Serial KoJ^Cl54 



(E) Proposed cougss of project ; 

1= Causes 'of death sxBong "loag teraf' survivors; m spscial 
study of 20-year survival experience will be Eaade, 

2o Reconciiiatioja of trend data on incidence, mortality, and 
. survival „ 

3, Probability of developing a second primary cancer » 

4o Survival fexperienca of patients that refused tres,tffisnt 
to evaluate the "natu:esl history of cancer ' » 

5» Detailed anslysis of the relationship' of therapy to survival 
taking into account the reasons for the selection of the 
prescribed course of therapy in each individual c®s©<. 



Part B included: ¥es 



24 



Serial NO. ^^^^54 
?3LVt B° Houoss, Awaxds, and Pi^llcaclons 

Publications other than abstracts from this project; 

Cutlex*, S. j.» Ederer, P., Griavold, H. B., and Greenberg, R. A.; 
Survival of Breast Cancer Patients in Connectieue, 1935-1954 
J. Hat. Cancer Inst. 23: 1137-ll5u, 1959. 

Cutler, S. J., Ederer, F., Griswold, M. E., and Greenberg, K A.: 
Survival of Patients with Uterine Cancer, Connecticut, 1935- 
1954. J. Nat. Cancer Inst., in press. 

Cutler, S. J., Ederer, F., Griswold, M. H., and Greexiberg, R. k.i 
Survival of Patients with Ovarian Cancer, Connecticut, 1935- 
l954o Jo Rato Caoser Zostoo in 



^aors and Awag'ds arelating €e this pr€>je@ts NONE 



25 



Decensbgr Serial Ho o NCI 55 

lo Biometry Breach 



PHS-NIH 

Individual Project Report 

Calendar Year 1959 



2o Demography Ssection 
3o Silver Spring, Mdo 



Part Ac 



Project Title: Retrospective Study on the Association of Smoking 
and Lung Cancer In Woiasn 

Principal Investigators: William Hasnszel and Michael Bo Shlrakln 

Cooperating Units: 63 institutions throughout the United States 

Man Years (calendar year 1959): 
Total: 
Professional 
Other 

Project Description: 

(A) Objectives : To observe ^ethsr th® association between ansoking 
and lung cancigr observed repeatedly for males holds also for 
females, A study among women will permit estimates of the 
separate effects of chronological age and duration of STUokings 
since isoaea o£ all sges took up smoking in the 1930 decade » 

(B) Methods Bi aployed; Histories of sraokiag and oth®r character" 
istics are obtained for each disgnossd lung cancer case and 
two matched controls. The cooperation of 63 institutions has 
be@n secured and histories are being collected by persons 
connected with each institution. The small number of lung 
cancer cases among woman make cooperative study arrangeffignts 
necessary, . ■ , 

(C) Major Findings; Histories have now been received for 290 
women lung cancer cases and 580 matched controls, much larger 
than any other series reported in the literature, A paper 
reporting the results for the first 192 cases, has been 
published. The findings ar© consistent ^ith other studies on 
woman reported in the literature in indicatiiig that the excess 
risk among female smokers is concentrated in the 
epidermoid and undifferentiated types. Also, the combined 
result® of several investigations suggest that, the character- 
istic excess lung cancer mortality among males elmost disappears 
^en nonsmokers are studied ^ since male nonsmok^rs have only 
slightly higher rates than f«mal® nonsmokers. 



26 



Fart A 



contlKusd Serial Ko,^£f_£; 

Signific anc e to the pzo^^&m of fc he ZnsSi tute ; S&& A= 



CE) Proposed eoagse p£ pro ject; Collection of histories is being 
discositinued; the extanded aeries will be analyzed for further 
infonaatlon on effects of duratioa of siBokitig on lung cancer 



27 



Sarlat Kfo^NG I 55 
T :-<:•: '■ doiioiTSs A^'jards, and pubiicafclons 

Publications othsr then abstracts from this project: 

Hasnszel, W. and Shitakin, M» B.: Lung Cancer Among ^IosrsUc 
Acta Unio, Internat, Contra Cancrum 15: 493- 501 „ 1959 



Honors and Awards relating to thle project : None 



28 



PHS-HIH 

Indi%'ldual Project Rsport 

Calendar Year 1959 



io Biometry Branch 
2 , Desaography Sec£ ion. 
3o Silver Spring, m. 



Part Ac 



set Titi®: Association of Smoking 'iistory, RssideKice History, 
and Occupation History With Lun§ Gsacsr Mortality 

Principal Investigators: William Haenssel and M. Go Sirkea 
(Hational Office of Vital Statietics) 

Other Investigators: Do Lovsland, I„ Mo Moriyasna, and Karl Tasuber 

Cooperating Units: National Office of Vital Statistics 

H&n Yssrs (calendar year 1959) : 
Total: 2°l/4 
Professional: 1 
Other: 1/<-. 

Project Description: This project is b@ing undertaken in coilabcv •. 
tion with the National Office of Vital Statistics ^ Division 

of General Health Services, Bureau of State Services « 

(A) Objectives ; To study the distribution of lung cancer taortality 
among population sub-groups classified with reapecE to smoking 
history, residence history, and work history » The iti'c&r- 
action ef thesa three variables in influancing lung cancg'^f 
mortality rates 1§ also b@ing inv^stigas'edo 

(B) Methods Employed ; Detailed smoking, residcncej and work 
histories are collected for sssaplms of deaths frotn relstivss 
of th® decedent and the attending physician. Additional 
diagnostic information on -iliich the lesdical certification qS. 
death was based is also gathered » Most of the data hav^^ bsea 
solleeted by mail questionnaires « Corresponding data on 
smoking, residence, and work histories for the gensral popaXati:. 
vmt& collected under contract ■»i£h the Uo So Bureau of the 
Census as a supplesssnt to the it Current Popuiatiorj Survey o 

Collection of data for deaths and the population base in . 
this manner peradts extension of the. range of varisblas for 
\diich mortality rates can be cc®put«do 



29 



?&tt k ■= eonfelnued Serial No. NCI 5^ , 



CC) Kator fladlnj^si Collection of blsfcoxies of a sample of 

1800 deaths in the State of Pennsylvania has been completed. 
This included the collection of additional diagnostic details 
on wiiieh the medical certification of death was based. This 
stiidy yielded useful methodological d.2.ta for the planning 
and conduct of further investigations. One paper dealing 
with the diagnostic evidence supporting medical certifications 
of death has been published. Other papers dealing with the 
methodological aspects of collecting supplementary data on 
decedents are being prepared. 

CD) Slgnificanee to the program of the Institute . Tliere are 
several imresolved issues in the epidemiology of lung cancer ° 
Several of these arc concerned with the interaction among the 
effects of smoking history, atmospheric pollution, and 
specific worlc e^osures in infli&^neing excess lung cancer 
risks. Studies of lung cancer mortality such as that dis'^ 
cussed here will provide useful itifstrmation for the planning 
of forward studies in urban areas thought to have sevctre 
atmospheric pollution problems. 

(E) Proposed course of project t The Pennsylvania experience was 
used to plan a study of a national ten-percent sample of 
lung cancer deaths. The national sample of male deaths has 
been completed. Histories for 2,600 males dying of lung 
cancer have been collected, as well as histories of 500 
mal«6 and 500 females dying of canser of the intestines and 
cancer of the rectvan. The collection of histories for 
lung cancer deaths among females will be completed by March 
1960. Preliminary tabulations of the data on male deaths 
have been tuade. Final tabulations and analysis of the data 
will be made in 1960. St is hoped that the findings can 
be prepared for pviblication in I960. Tlie Current Population 
Survey conducted for this study will also serve for an 
analysis of lifetime migration histories. Tabulation 
specifications for data collected In this survey have been 
completed. 

Part S included; Mo. 



30 



Serial l?o. NCl56 
Farfc B; Honors, Awards, and PublieAtions 

Sirken, M. G., Haensrel, W. and Pifer, J. W.s Reaideaee Histories 
of Deceased Persons. Milbank ifcmorlal Fund Quarterly (In preaa) 



31 



December Serial Ko gCI 32^ 



1. Biometry Bsraneh 

2. DeiBograpUy Seetioa 
PHS-NIH 3. Sliver Spring, Md. 

Indlvldujil Project Report 
Calendar Tear 1959 

Part Ao 

Project: Title: Studies of cancer mortality data 

Principal Investigators: William Haenszel and Tavia Gordon 

Other Investigators: Margaret Crittenden 

Cooperating Units: None 

Man tears (calendar year 1959): 
Total: 3 
Professional: 1 
Other: 2 

Project peserlption: 

(A) Oblectives : To Investigate possible applications of mortality 
data for epidemiological study purposes 

(B) Methods Employed s Special tabulations of cancer mortality have 
been secured from the Hational Office of Vital Statistics and 

a series of charts on regional variation in cancer mortality 
by site within the tteited States is being asses&led. A 
duplicate, set of punch cards for 1950 cancer deaths has also 
been obtained and the microfilm of dea th certificates have 
been reviewed to add country of birth of decedent to the 
punch cards Tabulations of cancer mortality by occupation 
have also been obtained from the National Office of Vital 
Statistics. 

(C) Major Findings ; There are significant regional differences in 
caneer mortality by primary site within the United States 
Mortality trends are also different for specific primary sltes^ 
The available evidence from cancer register and morbidity 
surveys in general support the Inferences to be drawn from the 
mortality data. Differences in caneer mortality uave been 
found among the foreign-born according to country of origin. 
There are also large differences according to occupation group. 

(D) Significance to the program of the Instlttite ; The mortality 
data thus obtained will be useful in furnishing leads and 
delimiting subject matter areas for investigation by other 
study techniques. These studies provide essential baselines 
for cantser investigations. 

> 32 



Part A - continued Serial Ko, mjr^'^ 



tE) Propose d cour se of project : Analysis of data for £he 

for@igti-bom groups in the United States i$ nearing coiaplfetion 
snd «ill be prepared for publicstion in 1960 .> Work is '^sll 
along on & systesiatic presentation of geographic variation 
and time trends in mortality for specific cancer sites , lliis 
material should be in a forat suitable for publication In 1960 » 
Present plans call for publication In Public Health Monographs c 

The systematic review of soorbidity and mortalit;^ data 
for soma specific cancer sites is continuing, with an .article 
on cancer of the esophagus being readied for publication, 
probably in 1960. Smnroaries of cancer saortality by occupation 
for specific sites will also be prepared in 1960s, intended for 
publication in the Journal of the Hstional Cancer Institute » 

Fart 1 included: les 



33 



Serial »o„ NCI 57 
Part ®: Honors, Awards, and Publications 

Fubiicffitions other than abstracts from this project: 

On Invitation, Mr, Haenszel has contributed & chapter on 'Spidemiology 
of Stomach Cancer" to Gastric Ca ncer, edited by Dr, Gordon 
HcNeer (in preas) 

Honors and Awards relating to this project: Son® 



34 



December- Serial .,,0, ______ 

1, BloHjatry Branch 
2o D@mog7S.phy and 
Cliaical Blonaetry 

P Sect ions 

FHS-MIH 3o Silvsr Spring, Mfi< 

Individual Project Report 
Calendar Year 1959 



Fart A. 



Project Title: Consultation on cancer registers and application 
of statistical mefchode to study of cancer in human pcpuls- 
tionso 

Primripal Investigators: Hilliam Haenszel and Sidney J„ Cutler 

Other Investigators: All professional personnel on staffs of 
both Sections o 

Cooperating Units; Hone 

M®n Years (calendar year 1959) : '' 
Total;- 5 
Professional: 3-1/2 

Other: l»l/2 

Project Description: 

(A} Objectives ; To assist other branches of the National Cancer 
Institute, State siad local hsalth departissnts, and other 
health sgencies by consultation and advice oa ti^thodological 
problems in the nsaintenance of cancer registers and on the 
analysis of data collected.. This includes the developtiient 
of statistical tools useful for the analysis of field ob- 
servations on human propulations and their application to 
specific problesis. 

(B) Methods Esaployed ; The conssuitation draws on standard statisti- 
cal and actuarial techniques and accepted principles for 
records manageiaent , ^th necessary isodifications required for 
an individual study situation. Many of the problems can 
be handled by correspondence, but field trips are required 
in several instances = Consideration nsust be given to the 
study objectives of the individual investigator or agency, 
the existing adiainistrative arrangements and the persoianel 
and financial means at hand to implement the program of 
operations snd studies o 



35 



Part A - continued Serial SiOoNGI 58 

(B) Methods SsEployed - continued 

Most of the consultation work ia this area concerns problems 
in the integrated bs® of data from various study techniques: 
analyses of raortality data, controlled retrospective studies 
prospective studies of special cohorts, morbidity surveys, 
screeniffig of populations for precursive signs, snd cancer 
registers a The Branch staff is particularly interested in 
the methodological problems and issues involved snd will 
continue to do research in this sre&o 

(C) Major Findings ; Acting in the capacity of a service organization . 
the Sections primarily assist other investigators in shaping 
their operations mnd forssalating their findings o Soststintes 

this aesistm&ce is acknoi^ledged in publications by the 
individuals assisted. 

CB) Significance to the prograiB of the Institute ; Many of the 
benefits of the consultation prograsa are long-r&nge in 
nature and consist of setting up record systems and study 
situations which enlarge the potential study resources of NCI 
and related organizations. The major activities in the past 
year are indicated belo'i?: 

lo Collaboration in the analysis of data frosa the Connecticut 
cancer registsr (this collaboration is so extensive th®t 
it is reported as a separate project) o 

2o Advice to hospitals on maintenance of cancer registers, 
a dessand stiisulated by the American College of Surgeons' 

prograsffl for tiaaor clinic accreditation,, 

3 Assistance to, the Cancer Chesaotherapy National Service 
Center in planning and performing analyses of the data 
reported to the End Results Section by cooperating cancer 
registries and in the davelopiaeat of protocols for special 
studies of selected sites o 

4, In cooperation with the Office of Biosaetry, Division of 
Research Services, further refinements isere made in the 

IBM 650 coEBputer program for actuarial analysis of survival 
experience of cancer patients o 

5o Assistance to Laboratory of Pathology in its retrospective 
investigations of cervical cancer asong Jewish and non- 
Jewish woman in Hew York City and Israel and of bladder 
cancer in New Orleans, 



36 



Part A - contiaued Serial No. WCX 58 

CJ3) Slfficilflcanee to the pgoggam of the In^titu&e; - continued 

60 Aoslstance £0 Dr. Zubs<t»d's Ssgistsy of tumor pAtlects 
treated at the Clinical Center in the technical aspects 
of maintaiaing the registry. 

7. Collaboration vith the University of Arizona Bureau of 
Ethnic leseirch in establishins population registries for 
Indian tribes in the Southwest. Such registrers are 
essential to the conduct of many epidemiological studies 
of cancer among Indians planned as part of the NCI 
extramural and intramural programs. 

8. Work on problems in the integrated use of the various 
study techniques: analyses of mortality data, retro- 
spective studies of diagnosed eases and matched controls, 
prospective studies of special cohorts, morbidity surveys, 
screening of populations for preeursive signs, and cancer 
registers. 

9. Preparation of professional aisd l«y edttcstion tsaterlals 
on cancer itiecidenise, mortali^, and survival. During 1959 
some fasts and flgijcres on the occurrence of cancer in the 
United States and other eountries were assembled for the 
Uo So Senate Conaaittee on Garvexmmnt (derations and 
appeared as a coimaittee print. Caaeer ; A Worldwide Hleaaee o 

10. Liaison with eaneer registers in Denma^g Sngland, Finlai^, 
Fraase, and Norway has been strengthened and an Ad loe 
Grotqp on International Cooperation in the Evaluation of 
End lesults of Treatment established with representsitioa 
from 6 countries. The Ad ]foc Grotjp agreed on & set of 
Rules for reporting end results and made arrangements for 
presenting international comparisons of survival data at 
the International Cancer Congress in 1962. 

11. A study of the relationship of adenomatous polyps to 
carcinoma of the colon and reetum has been initiated. 
Following meetings with interested investigators from 
other institutions a protocol and questionnaire was 
developed, a pathologic review board established, and plans 
made to collect follow-up observatioaa on 7000 polyp-free 
patients to determine the subsequent esperienee of the 
group. An instrument, the poiyposeter, was developed by 
Dr. Max Koppel whieh may lessen the difficulties of 
measxiring lesions observed vis the proeto- sigmoidoscope. 

12. Technical problems in establishing a register of persons 
with sxirgical plastic implants are being Investigated. 
Such a register would be an important tool in the study 
of body response to plastic implants. 



37 



y&ttmued 



(g) gropoeed course of project ; Special ateeatioa -will contiau® 
to be given to inteenatioaal collaboration In studies 
of survival experience and epidemiology with caecar registers 
ia England aed Males, Bemaark, Norway, Finland, France 
and the Soviet Union, 



Fart B included: Yes 



38 



Fart sr Honors, Awards, and Publlcationa 

Publications other than abstracts from this project: 

Cornfield, J, snd Haenszel, W,: Seine aspects of retrospective 
studies o J, Chronic Dis. (in press) 

Cutler, So Jc, and Latourette, H. £», A Natl-^nal Cooperative 
Program for the Evaluation of End R€«ult8 in Cancer o 
Jo Hato Cancer Inst, 22: 633-646, X959o 

Cutler, So Jo: "The analysis of long term survival experience 

ia humans o" Presented before Biometrlc Society Meeting, 
Pittsburgh, ?&. Ifeirch 20, 1959, 

Cutler, So Jo: "The use of life table asethods in the evaluation 
of ther&pyo ' Presented before American Public Health 
Association Meeting, Atlantic City, No Jo, Oct, 19, 1959, 

Hsenszel, Ho and Hillhouse, Mo : Uterine- cancer Morbidity in 

Hev York City and its relation to the pattern of regional 
variation within the United States o Jo Hat, Cancer Insto 
22: 1157-1181, 1959 o 

Hsenszel, W. : Some problems In the estimation of familial risks 
of disease o Jo Hato Cancer Insto 23: 487-505, 1959 o 



39 



Serial Ho„NG3>51 



FRS-HIH 

Individual Project Report 

Calendar Year 1959 



1, Biomatry Branch 

2. Exparinsental St&tietigg 

Section 
3» Bethesda, Mdo 



?.rt Ao 

Project Title: Statistical Consulting and Research Services 

Principal Investigator: Nathan Sfentcl 

Other Inveatigators : Edmund A, Gehan, Joan M<. Gurian, Lymsn Crittenden 

Cooperating Units: Hone 

ifen Years (calendar year 1959) : 
Total: 5 
Professional: 3 1/4 
Other: 1-3/4 

Project Description: Provision of consultative services in etstlsticftl 
design and analysis of experisaentSs \stth associated research in 
statistical ia@thodology o 

(A) Objectives ; The priis&ry objective is to provide assistance in 
the design, interpretation^ %sd svsluation of experimsnts «nd 
surveys conducted by researchers &t the Mstional Cancer Institute , 
(and other institutes, on request). This includes services as 
^n associate editor for the Journal of the Mation&l Cancer Institute, 
(Mantel) p v&rious referee and review services, and n^thematical 
assietsince for development of theoretical taodels of biological 
behavior, 

(3) Methods Eatployed : The mssistsince described sbove is sccomplishad 
using the stimdard methods of bioisetry, statistics, %nd pro- 
bability j with the necessary raodificatioss required by the 
problem at hsnda Frei^uently, the standard statistical stethods 
ars not sufficient to handle the problem on Tshich assistsace is 
being given, mid it becomes necessary for the personnel of the 
Section to originate new s^thods^ 

(C) l^jor Findings : As a service orgsiQization, this Section primarily 
assists other resssrchers in tsmking iiaportsnt biological snd 
medical findings 5 rather than la making such findings directly,. 
Such assistance is frequently ackaswledged in the publication 
of findings of the researchers o Esaaiples of the kind of research 
problems in «hich assistance is given are shown belo^, Occssiosally, 
th® nature of the assistance is such as to warrant co-suthorship 



HO 



rather than simple aclmo^ledg@isent , Also, ne^ et«.%ieticml 
mathods originated in this office to handle certain probiema 
&Z& Boeaatirs&s trnpottmnt «nough to warrant sepsrsita publication, 

(D) Significance to the program of th a Institute : The significance 
of the contribution of this Section to ci^ncer research is b«st 
illustrated by the probleias in ^ich the Section has provided 
statistical &8si8tenceo These include: 

(a) nutrient re^uiretaants of cells ia tissue culture; 

(b) carcinogenic properties of chroasiuia coi^ounds; 

(c) HKjdels for threshold activity of a carcinogen; 

(d) infliience of oxygen coacentrAtlon on radiotherapy of tumors; 
C®) toxicologic propsrtlss of specific antitumor agents; 

Cf) induction of tumors by 8ter<jids; 

(g) studies of snti-leulKeBiic agents with respect to sffic&cy 
and toxicity to host ; 

(h) investigations of possible synergistic coiabinations of 

aati-leukefflic agents:, 

(i) screeaiag programs for ant t-timor agents-, 

Cj) prcgraras for assaying hormonal effects of steroids; 

(k) epldefuiologlc studies of csncer; 

(1) studies en the incidence of lung is^tastsses folloiJlKg 
surgery or other treattaeat; 

(sa) assay of chicken lymphoraiatosls virus; 

(a) study of genetic and othsr differences in response to 
cheaao therapeutic agents; 

Co) study of inter- relationship of factors influencing 

i^tabolism in the aon^l and diseased rat; 

Cp) evaluation of therapl®8 in controlled clinical trials 
conducted at NIH; 

(q) metsbolic studies of patients; 

(r) studies of caleiura and stroatlOTi incorporation 



41 



rt A ~ continued Serial K^.N GI'-° 51 



It is anticipated th&t in the coming year the degree 
of sxtraraural collaboration ^th cancer investigators «ill 
increase = Such collmborstion this past year hss included vork 
xiidth investigators at The Atotaic Energy Coimtission and the 
S loan«Kettering Institute , 

Following the trsjssfer of Dr., 6eh£n to the Therapeutic 
Trial Section, CG£ISC, the Exparlmsntal Statistics Section 
discontinued sctive consultation to clinical groups aseociated 
with the CCHSC progrssm, since their needs could be aset by the 
statistical 3t<%f£ of GCNSCo 



Fart B included - Yes 



'42 



I 



Serial l?o.NQ^i_ 

S&vt B: Honors, issas'dSs aa«i Fablicatioas 

PisblieatioQS other thaa abstracts froa this project; 

Mantel, M. £nd Eaeassel, W. : Statistical Aspects of the Analysis 
of Data f£<»j Retrospective Studies. J. Nat. Cancer Inst. 
22; 719-748, 1959. 

Goldia, A.; Hsss^hreys, S. R.; Vendittl, J. IS aad Meatel, K. ; 

FroloagatioQ of the Lifespan of Mice ^ith Advanced Leukaisla 
(L 1210) by Treatment with Halogenated Derivatives of 

i^ethopterin. J. Mat. Cancer last. 22s 811-823, 1959. 

letchm, A. S.; Wsxler, H.; and Mantel, K. s The Effeet of 

Reaaoval of a "Priaary" Tumor on the Deveiopaent of Spontaneous 
Metastases; I. Developaent of a Standardised Experimental 

Technie. Cancer Research 19; 940-944, 1959. 

Goldia, A.; Veaditti, J. M. ; Mine, I.; and Mantel, H.: EvaliaatioR 
of Aati-leukesic Agents Sa^loying Advanced Le»fcesaia, L 1210g 
in Mice. (Cancer Chemotherapy Screening Data IV) Cancer 
Research 19; 429-466, 1959. 

Liedaraaa, R.; Douglas, J. 0. A ; and Mantel, li.: Production in 
Mice of Ascitic Fluid Containing Antibodies Induced by 
Staphylococcus or Salaonella Adjuvant Mi3£tures. Journal of 
>logy. In press. 



Goldin, A.; Venditti;, J, M. ; Euaphreys, S. R. ; Shuster, L. ; Darro^, 
a. A. and Mantel, M.; Advanced Leukesiia as a Tool for Ohes^- 
therapeutlc Studies . Acta IMlo Internat. Contra Cancrtns. 

In press . 

Mynder, E. L.; M^tel, M.j and Licklider, S.s Statistical Considera- 
tions on Circumcision and Cervieal Cancer. American 
Journal of Obstrics and gynecology: la press. 

Honors and Awards relating to this project: 

Nathan Biaatel has been invited to present a paper at the forth- 
c<»&lng s&esting of the International Statistical Institi^te 
at Tokyo, Japan ^ in 1960 



43 



MTIOTAL CANCER I!?STITUTE 
C&HGE51 GPiSI40THERAPI MTKMAL SERVICE CEOTEIR 

Cancer Qaanotherayy Prograia 

ISas nstioxifil program of researoh for drugs for the cHeSLotLL 
tz^atment of ^«oer has* entersd isfto tha fifth year of operatlona 
Hajor effort during the yaar has bean dLreetod on reachisg goals 
set for procureiQsnt of nev ssaterials^ pro9idi2»g for tha screenlog 
of these nfiterlals for potential actlvl^ i^jaiiost cancer and for 
^t& €oq»mslon of cocper&tlTe tfLlTrfcal gzocqps to evaluate tiis nev 
isaterlals dlsplAjdng proozLslng thdr^>eatie actlvit^o 

T!t» greater part of the aetlvltlos of the Gentor are 
carried out ^ eontr««tso ^ese cootraeta are vltli pharoaceatical 
and other lodtustrlaL c^nems, eoonerclal consulting laboratories^ 
universities and aoa*profit ox^esalMtionao Coatreeis are utUlaed 
to procure defined areas of research aetlvlty- fl&d include 
acti-Titles such as drug procurraaent end tAm^csLX STnthesis^ 
evaluations of drugs for antitwaor aetLvity^ search for hotter 
aethods of drug evaluation^ and pharstacolsgy and toxlcolog^o 

CsKitrscts are also utilized to collect statistical dmta. 
from a group of esncsr registries representing a varieV of asedical 
institutions in irariovur parts of the United States » ^is retro^ 
^pecti^e statistical program is designed to deterEsine the natural 
course of the disease and to find the response of cancer patients to 
Various types of treatment laaasured in terms of survival » Data 
ect& being collectsd frcsa lit individual registries and plans are 
being dev^eloped to bslng other registries into the progrsa 8i»i 
institute collaborative studies ^th selected Europeasn Registries » 
In additicasp to ^et the problems of central centrol and 
coordination, contracts have been established for ^s statistical 
design and servicing of clijiLcal evaluaticm trialSo 

Coopsrativ© agreements have bean entes^d into with other 
Federal agencies to participate in the chemotherapy programj 
ineludingf the Food and 2rug Mministratioa for pharraacological 
and tozicological studies, Departssent of the Anay for eyn1±iesis 
of coEspoundSj and t!^ Veterans AdMaistratioa for clinical 
evaluation of chsffiotherapeutio agents o 



44 



2 = 



Additioually;^ TOsearch grants ar@ utilissd to casry out 
a portion of the di£=@cted prograiBo fhss® gj^ants at the ps^esont 
time ara largely in coojtssetion vith the clinlc&l tesldiig of 
dmigs by cooperativd clinical groups o 

The Hatioaul Advisory Gancar CoundLl^ fi.t ita June 15»17d 
190?y ai®8tizsg« recoisnendod ecoeptanee of the follosisg guid0<° 
lines for contr&ots or grants in sxipport of rasearch and related 
activities s 

The aozsial ehoice for support of resoar^ Kill 
he th® grsat astibaniSB^ for it p(ro7ide8 tte fireedoa 
of fte^oa t&i&iAi is easwatial to seieotlfie peoenvs^ 
OoQtraets way hn lUMSd vndar the foUesaiag oircnestsncess 

(1) fbs eontraet relatds to a psrogr^ fMdt baa li^ea 
desigoftted l^ tha Director^ Kfttienal Cancer 
]&a8t£tiite, as teing one for iriiicb osa of eaa» 
tracts is desirable and for ubich ftods hava 
h^n speoifioally aansrlsedo 

(2) fhs psrograja offioials td.th the ed?i©s of apja^p^ 
riate advisory gros^ define specific &r®&a of 
aeti'^ty %?hieh siay be sv^jported vis.&&r eontracto 

(3) Gcaitrsets lisust Kset program ©bjscti^^s as 
dst^radaed by offieiala of l^e prograa areag^ 
most .\3& idtfain lisdtatiosss oi funds^ detenaised 
to be substantially soondf sad eontribcste to a 
balanced program ±n the opinion of scientific 
advisors and staff of the progrsp areao 

(U) Th2 contracts to bs considered ^11 t® entered 
into vsgan invitation of the program and designed 
to iseet the pro-am needo Xs^titution@ ^hich 

id^h to partidpat® in this program Bay also 



!Ehe dollar support of Mt© total "^rogrsKEsed" acti-^ties 
of the Center ai^iears in fable !<, A cossplete listisjg of con« 
tracts^ allocations of fimds to verioos research areas^ and 
amounts transferred to other Oo?ernment agen(d.ea appear r@» 
spsctitely as App^idiees I» II j and HI*, 



45 



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1!t» aetl^tiea of tte CCItSC havo d^nralopod in tha 

Cfae8Blerta y"«»d3!nig gyntheals aad Tprocuronent o Synthetic oheodcals 
'^^'Is^^SS^J^rS^SSSriSS^noSjSgv^^'VBd at tbe rate ^ 
about liO^OOO anmnUy for introduie^xm into tlia primary sesweUo 
"Biese ioaterialfl ave ev^iplied Isgr ohsndcal eynthesis contg«etox« 
aid "prop lab" ewtraetors i^ Svaraish ctaBsereiaSLljr iiaobtala<» 
abLs eao^oiandss and vslversltlea* rasoaroh op0)ad.sE&tlQn8» and 
pzlvatQ piiaxss8ea«U.«tdL and dmg eoaoasnui ^lieh vauaXly funai^ 
the matffirialn C^reao 

to additl<naal 500 QT&tbetlo nsitteerlals per acmth wlU 
bs obisdEned for seroenisg in tissua etfltaro tMa 3ear« desa 
a3K» mftt@zial8 th&t batra not bean tested -pTmLoasly beestaaa tiusy 
are STallabla onl^ in ^razsr saaU (^aantltias Insaf fioiaBt fmr 

testing In tbs aninal sore@ns« 

^^ se^eening of plant products has rscelvad little 
smpbasis in the pasto Sa^«?al such isa'teriale that ha^?s l^en 
screensd baf® axiiiblted intsrssting aetivityo It is proposed 
t€j take advantage ©f ps^Tioiis work iss tliis field 1^ -^isrloiss 
instituti@ii3 aad obtain plant ©sctracts at th® rat© of s^sfs-sffsl 



Isrga supplies of naterials that ha*^ displa^d 
"aetivligr* in the screena. are requtrad for pr«oliid.^l tgssieity 
tasting snd phaa^aoologieal ^$ork°^^ aiidj, additionally^ n&w 
agents l^eoadng sFailabl© i^iiirs prodTiction in siaf ficisat 
quantities for clinical ©valuation, GoaseTosntly, contracts 
for chemical synthesis and "prep labs" ar© being expanded to 



Primagy Screening ■?■» Aniaial Product ion^ The basic scraen of 
the Center canaists of tests for activity sgsiast fotir sous® 
tuBiorss Sarcoma 180, Adenoearciasrca 755* Laukezaia ISIO^ sad 
an altemat® added this year^ the Ehrlieh ascites tusior,, 
With th® aetsblishiseat of "in planf screens in industry, 
scresaiag capacity of th® Center has now ?©sehed a Is^el ef 
about 225 jOOO testa anaisallyo HovsTer^ tb® original estiinata 
of lioS tests per emipo\m6 has been found to be too eoaser^ati-?® 
in relation to testing an-tibioticso A more accurate figure 



47 



falls l^tn?@en 6 and 7 tests p«p "beer^o 'Ebis v&b ye-^aled ia 
res^re^iing actl-ve "beers" after fractionAtion to diaelose t!^ 
£ctiv« iagredLent» fiRsad on this new figare» in order to prth' 
vide adaquatd tests f(n> liO^OOO cegapoxasdB annually^ additional 
screening projeets will be addad to provide the level needed. 

An iii9>ortant developtaat in the aer^miag program 
occurring dcols^ the past year has been the utilisation of a 
fully nec^ianlsdd data processing maehlneo %!& Tfill malse 
possible critical reviev and evaluation of the great nase of 
data aceuRBilating in a substantially shorter period of tiiae as 
veil as famishing a quality control ne8»ure to assvore eoa^ 
sistent test z«sult8o 

VWi the g3roHt^ of screeninig ea|3aei1gr5 it -was ia^)erati'90 
to increase t&e production of nice and rats of inbi?ed strains 
to meet requireraents of this progren. 'Bm sxsppLj has been in<=> 
creased fTcna & level of lOO^COO to av9t l^SOO^OOO per ^aro 
Standards for sound sniaal production and diSisaee control hsva 
been established to assure ecHatinued production of hi^ quality 
laboratory aniioalso 

Secondary Evaluation «« Meth odology » At the tiiae the broad 
prdraary "screen •was" es'SabHaiJ^ti^'' '■^Ee screeEsing advisoxy panel 
expressed great interest in sattiag up secondary ©valuation 
systems <, It wta expected^ based <m scienti^e conelusio^iSf 
that iBore unique zmterials laight ba uncovered which could prove 
raore usefiul clinically than substances eus^ently being fetaad hy 
existing sethodSo In general^, each secondary system new used 
is intended either to suK>leBsant or su5^1a«t the present pri« 
isary screen^ Efforts are nos? t^ing made to assess the s©c(mdasy 
evaltiation projects now in erlstenceo The (smcial t®st^ vis, 
correlation with responses ia iBan, remains t© bs ®s-fcablishedo 
Intensified activities in tMs area have been strongly urged" 
by advisory panels and additional projects are now being developed 
for this purpose. 

Secondary evaluation has proceeded along the follo^ng 
lines: 

^s IE ^^19, techniques 5 involving th© use of cells or 
mieroorgsnisias as a quick test to evaluate anti- 
laetabolitesj for use in studies in cross resistance 
to drugSg or collateral sensitivity studies o 



48 



2o The i2se of a largs vsriety of «miisal tumors as test 
aystesa in ordar to ddvelop lE>o&der biological data 
about materials scrssnsdo 

3o QETosiing taaaan neoplaoos In a Tariet^ of hosts or 
in ^tgo in orddr to dstosiaine irtiethsr such test 
sys^eiasljavB greater predictability for nsng 

Ai^roprlate Clinical data are not yet on hand to nake 
reliable assessoisnta of these systeias. Efforts are eonstantly 
being made to find better and less e^qpoosive aothods of screen- 
ing drags. A review of these afforts is nov in progress to 
detexmine idiat ne«r direction^ if any, should be talc!»)o 

gcadoology and Pbarga«)iogy » Adequate facilities for precUnieal 
toxicology and pharmacolog^ ara believed to be a:7ailabLe to 
satisfy- the present seedo A good sat of working protocols has 
been established and is providing clinicians vlth necessary pre» 
clinical infonsationo Hc»re'7er, clinicians have receatSy expressed 
an interest in the dsrelope^nt of a protocol on coiaparsitiTe 
pharmacology of related agents o This is no^ being considered and 
raay reqidre expandsd facilities to obtain approprlete infoziaationo 

EndocrJnolegjTo 1h® endocrinology prograia has rscelYsd extensive 
Sn^rss^" ai^Bing froiB a great boc^ of scientific fact and theory 
relating honeones with cancer., Approaches to the solution of the 
cancer problem based on these v±@wa ar© considered necessary and 
of great potentials 

Basically^ the porpose of the program has been to f^imish 
workers in the fields of sssperii^ntal snd clinical canc@r td.th 
mesms and infoxmation for nsing ne%r endocrins^related compomids 
in their studies o For this reason a search «as organized for 
c<^^>ound3 id.th hon!K>nsl or sntic^hortoonal properties ^ or ability 
to alter the functioning of an endocrine glsnd^ so that U^ 
G(MSQ ^ould be in a ji^sition to furnish the clinician or 
experim^italist ^th compounds that laight be ns@f^ in testing 
a working hypothesis o Secondly^ in order to characterize -^ese 
cmpoisids physiologically (as iatp^rtsnt as their chesdcal 
identification) for the user^ a bioassay progrsii "Sfas set upo 
Finally^ in or<^r to retract froia aniajal timsr systems the kind 
of infoziaatlon that t^uld bs usefiiL in such a specialized pro» 
gram^ a search ^ras instituted for @ndoerii5©«relat®d tuiaor systess 
for use in testing these cc^>ound3o 



49 



Tba great and interesting task of exs^rdxig and snal^iag 
the irealtb of d&ta BCcoBralatlng Is natt going <m and is oine of ths 
Biajor goals for tise y&sPo 

Sie establl^biasnt of "prep labs** has greatl7 sitfianesd 
results in tMs program. Prior to the tiins these contjraets s&ra 
set up, the supply of materials for laboratory and clisical toting 
vas very limiteda At present the Cancer Oieraotherspy Ifiatlosal Ser« 
vice Center is furnishing 53 materials to 66 senior investigators 
engaged in clinical trials, Tob '^ep labs" supply an iisportant 
portion of these materials « 

The endocrinology program has been set op to bnild a large 
isv^xtory of compounds Kith endocritte-iAhlMtory activity^ to 
characterise these coB^poxinds in as many useful aspects as possible, 
and to offer thera for eaq;erinsntfll and clinical use in c<s!t«ct with 
an abundance of inf oxna^en that soy be t^ material of ne^ working 
hypothesis o It is ho!»d that this approach will increase our 
understanding of endocrine factors in cancer at the sasie tinse that 
all possible speed is achieved in actually finding endocrine agenta 
useful in treating cancer, 

ClJaical Trial® ,. The clinical program of the Calmer (Siemotherapy 
flai^onal S«hrice Cantor is conducted by cooperative clinicsl 
groups in varioi^ public and private hospitals across t&@ nation 
and in the fetas'ans Administration Hospitals o Groups other than 
the Veterans A<^2;d strati*^ groups, are su^>orted hy research 
grants. At present there are 17 gro*.^ participating in this 
program^ representing 1U7 institutions o A chart iS»3«ing the 
categories of i^oplastie disease being studied and the c3ie!ao» 
therapeutic agents tised appears as Aj^ndix 17 to this report o 
Cooperative groups are no^ studying 109 coB^pounds, elassifled 
as follows s 



50 



8 « 

CSLINICAL fSSIS OF COMPOOT^ 



coaigKnindis 

studies 

e«i9ileted 


Used 86 New 
standard caapovnda 
of telng 


Total 


9 


9 35 


53 


l 


5 13 


22 


3 


1 k 


8 


li 


h 8 


16 


1 


2 7 


10 



SteroJLds and ^x^ionssoaoaoeroo 

A31c7l&ting Agents c^o 

Antibiotics. •». 

Antiinetaloliteso. ..••. 

HlSCSXlSneOQB 09*o»s«aa«a»«a*a 

Total a 21 67 109 

Flans a?@ b®isg i^tls to sicceles^'^ 1^^ testing of drugs in 
Toasis for dsnj-ga ar® beeondjEJg rsady ia inef®asiag naaSwrSo Since 
Jaiiiaary 1^ 1959* the J?aw Agents Cosmitt®® ©f th® Clinical St\iai©s 
Panel has passed 10 agasits whieli ha-?® shoMn sufficient animsl @nti<» 
tumor ffieti^tj" to ^mrrant hisssm trial assd hs^® had adsqtmie pr@« 
clinical phajroacologyo At present 17 other nsw oozspounds are in 
dif f ©s?©at phases of praelinieal phsnsacol^gj or e^^>ar&ti7@ therapOTtic 
trials snd should bs ayailable for hsmmi studi®a soasatime in th« nszt 
six to t»®l\r@ iQonthSo 

A related and more difficult problsai facing ths progress is 
the design of clinical studies to secure data isbich Hill enable 
modiilcEtion of priinsry screens for batter correlation botween 
results in animals and in mBxio To accc^^lish tMs^, ImT^diate ®x» 
pansicai of the cooperative clinical group eapacitj for the evalisatioa 
of antitumor agents has been recommended <, ^e cat©gor5.es of 
malignant has^n disease to be used to evaluate this problem are those 
in which some of our eurrsntly aveilable chssjotherapeutie agents hav© 
some deiconstrabl® effects o It is anticipatsd that these vill utilise 
the available clinical material in tfees® partioular disoase 
categories for a ysar or aoreo Ths studies now being planned ar® in 
the follo^»ing mUgnant disease categories: 

(1) Acute leukeiaia in children and aeate sa^iogenous 
leukeada in adults » 

(2) Chronic lymphocytic leukemia, Igragjhoeytie Ij^hosarcosss 
and reticulum cell sarcoaao 



51 



(3) Cbr^iie i>(5r@log@£ious le^ikaada. 

(ii) »9dgkins dlsaasd* 

(5) Rhabdra^sarcosia a&d ^ln<8 ti^ovo 

(6) Netsroblastoroa, 

These studies trill sreqiilrs enlfirgenii^t of otir f&oilltles 
for e-^aluatlJtm of xuar naterislSa 

Of our p^ressn-t cXixdcal groups, Vas Stirgle&l A(3j%vBitt 
gctnqps are e^idbactiag their stuidles of surgery ceoibixidd vitb 
chataothermp9r» utl3isBiskg agents of estabUedbed value o TbB Prostate 
aod Brsftst QrotQKS ars using primrily stezoidSo Five groups ar® 
eralYLstias was agaats tliat result frooi aaia^ scresiidLag vorka 
These IsiBt g?£wspa ere not snffieleat to aset the growing Tolaaie 
of drtigs to b@ tested and loore ijistitiztiona have been inelted to 
subsdt grant appHcatioi^o 



The Csmeer Chss^th©rapy Satiosa®! S®r^c© G©nt®r -Hill ha?® 
reached th® first stage ia 1*ie develops^nt of th© national ch0isiG<=> 
therapy progrsaa early ia 196l| io©op b®w drugs b©ing developed 
and evaluated at a highly tsatisfaetory rat®^ and !;^ohin@ry 
established to process active materisls through oliMcal trials 

The nest stage will evolve from a cosiplet© asssssment of 
the eadstiag operatioBSo Every phase of th© program is uader- 
going a re»evaluati<m 1:^ scieatific panels ©f e55>srts based «poa 
ctaisiderations of results achieved^ ad^raneeiaenta imde, and- 
validity of research designs « Recosansadatioas of these panels 
will provide bases for the dstemdnation of research diarectioa and 
plans for the succeeding years o 

These plans say inelads one or more of the following courses 
of actions 

lo Intensification of consideration of problaias of 
primary and secondary evalij&tiono For , esaiaplej 
espansioa in areas of evaluation stich asi use of a 
larger spectmm of j^piirml tunsors in ths screen, the 
use of hosan tussors as test systeins, or t233 of 
cells in culture or of adcroorganisms as a laeasura- 
js^it of groirth inhibitions o 



52 



1 



10 



2« Iner@as@d effort in the ioXLmt-^ of exlsl^ng IeadS£ 
possibly including escpandsd facilities for ol^taisiQg 
preclinical iiifornuitlony cOi^)83r&ti'«B pbarmacolog^ of 
related ag^ats, or eat&blislsBent of additi(»aX 
fadHities for the stud/ of drugs in nano 

3o ReductlMi of enqphula in any of tbe fotregoiag activities 
in ord^ to escpand areas dBoonstratiog greater potentialo 

ko Establlsiuitent of addLtlooal oliidosJ. gro\^8 to test 
tu(9 6eags that bave becone a'imilabldo 

Before aqy nstr plen tdll be instituted^ careftOL eon» 
sideration uill be aads front the standpoint of iajutot upon the 
research cooinmnil^o Care "wXU. be taleen to prsMi^kt dL^rarsicn of 
scientific end technical staff from oqasaiXls iaportaat areas of 
researdb or education and to prerent the supplanting of othsr 
research fui^dse 



53 






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00 


w 


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s 


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-.4 


9 


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3 


> 


4J 




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to 


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O 


>. 


r<4 




4J 


ea 


• 


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.a 


U 


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W 


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g Ai 



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66 



I 






Was® of Qeo^ 






Greap A 






Fhas© 



11/" 



6»MP vs, ^^W « AasseslEiaa 

6-MP at -^sriotas dosagss vso 
6«^ * DOH. 

Steroid indtsosd reiaiasloas 
tz^ated with 6»!iP o(£> 6»HP 
end DOHa 

6-MP vg« Thioguanine va* 

Thi0gUSSI93iQ& e 

S»FU (prelisiaary)* 

ietiaorr^ia "Bo. 
Mitess^sia G 
5«?FI3DE 

6-M» Ribosld© 
Hipaeil B 
Asa'craeil 



lU 



Aeuts lf®?ite^© Aeiat® L®?ak^ia all 6«MP * @s^^opt®siB ®t 

6 Assorasil vso 6«.I# ^• 

6=.MP effects oa st®roid 



12 



67 



• 2 - 



-gLJ^BPu P^?^,^®,®(^,1 ^,'fep^®4 , 



CheiaotherapeutiG Agents Hoo Hospitals 
Used ^ .F^^^S^^MMl 



Peoled Lpiphoma 






Phase I & II Studies CytcBcan in Aeuts Le^kesaia 
Actinos^dn D ia Wilm's 
ToiBor IfeifehylglyHssai'-biS" 
guaaylhyds^stoii^ 



Brsassl; Group 



B2^3ast CfircinoiBa 



Testosterens propionate yso 
29 other steroids and 
hormones 9 (9 coHg^latsd) 

Hsloteetia vis, 6 o-^i®? 
hormonal stibstaacsse 

Thyroid ®3et?aet TSe plaesbo 
in castra-feed patients <> 

Co^tixd.soii of pr^disisoBs^ 
©thynyl estyai^ol sssd 17« 
xasti^l t®stost®roa©,o 

Progestoronsj 9a«fere^3<=ll»» 
fc@to vSo pxdoi? 8sp®ri8nc®j 
pr@dBisoB@ or (^tlsiusQ c&x°@& 



32 



Eastera Solid 
Tuffior Q?oi^ 



E©dgM.as 

Foolsd L^s^shoiBS. 
Misltipls nsBlmSi 



M2 ■3'So, fhioS)PA (lot? do3e)-5s- 

HIg rso fhioTSPA (high dose) 

DichloroffiethotsisjE&t© vso 

ffetliotrexat® ysa HI2 



AzaseriE© vso optiisam ear®^ 
^enchogsnic Osyeisisssa M2 vso aioSSFal* 

EM2 ■«- l«pa5r TSo I-^ray alosae^ 



68 



. 3 - 

Cheiaot'mrapeutic Agents Ho, Hospitals 

g ^ of Qr oup DiaeagQ(s) Studied gs@d ^^ PartlcipaMng 

Hslanoiaa Mg TOo fSiioTEPA* 

HNg vSo ThioTEPA 
Breast CarcinoiBa ffl2 to- ^OaioTEPA* 

BOP vso Prednisolone* 

Phase I & n Studies 0«FU 
A«139 
Dichlox'oiitsthotrex&te 

Karcotine 

Uracil l^stard 

Methylgls-o^d-Ms- 

gu8aylhyd?&2on@ 



Midsjest Group Adult Acute leubsmia 6«MP vso fhioguardiie -^So 18 

ThiogoaKosins 

Hodgkias Disease Cytosan vs»' Nitrogen !&istard 



Prostate Cancer Prostatic CareiiioiQa Placebo tso diethjlstil* 12 

Study &*ou,p l^strol 

Diethylstiltestrol (5 sEg^/d.,) 
vs, Biethjlstilbsstrol 
(500 tsgjdo) 

Metbylstilfcestrol vso testes^ 
teros^e propionate 

Phss© I & U Studies 17 ass? hormonal eos^unds 
in Prostatic Careiaoma 



Southsastsra Gbroais Lysphocyti© GhXarasbucil vso Hylersaa 

Group l^ukeiais 

(Jhloraabucil vsc CB-l^U 

U-782li (prsliMnary)* 



69 



I 



»li» 



Haae of Group MagaseCs) 'Stodied 



Gheraoth®rapsutic Agents Koo Hospitals 

Used ^^ ^ ^ Psafticd]pg.td8ifi 



Ghz'CTi.G IJ^Xoc^lc 



Pooled L3?mpho!!ja 



QjloraHbuclX g'So P«38 'Too 
cosi'blQtioiss tfesyapy)© 
Ghlorasbi^l vs« Ji8ylea?an» 
Cblorsmbueil 7s« CB»136U 
U=782U (proliiaiBary)* 

Chlog-asibaidl tso l^laran (abandoned) 
ChlcKPSEbKeil * pipednisona 



CM.0PS3sbu6l3. ( aban doss d ) 



Bresst CsffeiBOsm 



Chlorambucil (abandea©d) 



Phass I & LI Studies BOM 

GB»136lt 

HSC-.1026 

U-83i^ (Uraeil laustard) 

PA-liiU 

Uw782U 



S©«t;lw©st GsncsE* Aeuts I@ii&©i^a in 
Ch©isa3ths3?apy ebildr®n 



ACTH * hydroeoptison® «s@d 
alosse snd in ssquenes Mth 
6-HP 



ChroEie Ljmpboeytic ChloraaM^^l t3o ?22 



6-!®' ^So Myleraa 



Chronio f^?elos3rtis 



Phase I & II Studies Mlraeil D 

Csrainophilin 

HSC-1026 

Harcotine 

CblorGq.ui3!3 laistard 

BCM 

ActinomS^in D 

Actincmisnein D ••• Cytoxan 



12 



Name off Gyoup 



-5 » 

Cheiaothsra^utie Agents MOe Hospitals 
DlseasQ^s) Sttidied ITsed Fartieipatiag 



L«i*h®nylalanias Eiustard 
U-(p°I)i3!athylatniKostyryl) quinolina 

GytGsaa 
mtoja^ia C 
5-FU 
AB«100 



Surgesy Adj^avant ^®asi; Carcinosis 



ResectioB * fhioTEPA vSo 
Resaetios * Placebo 



13 



Surgery Adjufsnt GarciiioBa of Coloa Resection ■»• ThioKPi vs, lli 
Colon Growp resscticai alone o 



LuBg Gro^jp alone o 

Surgery ■*• Mp r&o surgery 
* placeboo 



Surgery AdjuvBsat Carcinoma ©f Stomach Ressctioa ■«■ ThioTEPA rs o 
Stomaeh Qrovsp Rssectioac 



StJTgary Adjir^^at Csreinosa of Oms^ Chloraabucil 
Ovss?y Qtgv^ 



2$ 



?©t®r®as A^iai©« Acta-bs Le^mla Adults 6»MP ts„ 6-CP 

trati(ffis Gh©ffl©th®s=&|:^ 

Q^mp Hadgkias Biseas® M2 -^o Iiti?oi!sia«- 

1^2 ■ysa Cytossa 



11 



71 



=» 6 = 



lane of Qroup 5iB8a@@(s )^^ Sttidied 



Csed 



NOo Kospitsals 



Peeled hjmph0Ts& 



Lmsg Garciiwssa 



M2 Too DOIHi- 

KI2 ^®» Cytasan 

HSI2 ^o IX)IS» 

Mg Too DOH'fJ 



Phase I & II Studies Gyto^a 
NSG-1026 

A-139 

6-HP Eibosid© 



Vetspgns i«MMs= G@?ei3io^a of Qolon 



Csi?eiB9ma of Long 



Surgery <■ 'OaiofSPA 
Siargsrj •» ?ls.ssbo 
S^J^gerJ Aloss 

Surgery * Plaeebo 
S'urg@s'y A3.0RS 
Gareis^a^ of Stoiaaeh Stirgary •> ISiioTEFA 
StK-gery * Piacsb© 
Syrgery Alons 



21 



72 



ChsExothexapexitde Ageats Mo* iioapit.t 



Veterans Adiainis* Broachogesic Carcinonm HMg ■*• Testosterosj® 
tration Lisng propiosat© ts 

(Honsens) Group tso progssteroae ve* 

diethjlstilbestirol vse 
inert eonipmrnd^ 

Testosterone proj^onate vs» 
Delta^l-testoiolactone vsa 
Halot@stin "rso HN2 vs. 
Radiation vs. inert corapo\saS 

Phase I & II Studies Cjtoxaa 

Uracil imstard 
Tetrefflia 
Miton^^n C 
AB»100 



Western Group Chronic l^miphocytic Chloreiabueil tso TWi 



Chronic ju^lecytic Deiaocolein ts» l^leran 



11 



* Ghloraxabucil vs. 



Foolsd Jj^s^ixms. 



2 * piaesDO vso Hsjg 



Prsdaisoas tso Placebo 



*Studiss Coaspletsd 

Total nHiab®r of noa pfeas© I smd phase U studies ^ 139 

Htiaibsr in progress - 85 

Nmber cosapleted » h9 

Htssber abandonsd ^ 5 

Number of group |tese I and phase II studies ^ 70 

1/ Preliiaiaasy glinical seti'^ties to obtain (Phase X) tosieological and 
"^ S^anascological inforasaticsi and (Phass II) data on thsrapeutic raspons©^ 
optiaiam rout® of adsinistratieng and optiEoaa dosagjs r©giffiSo 



73 



Field Xis^eselgaSiona and Dentonstratloas Braach 
12 Moaeb Period 
Repore f©r the Period Jamiuiry 1, 1959 through December 31, 1959 

The mission of this Bramch is coEacented with the acquisitioos 
of stew knowledge about maligaasit diseases through isiivestigatioffis of 
their i&ature, clinical course, aaad occurrence in humasis and the 
utilisatioim of such findings in the development of methods and pro» 
cedures for the identification, diagEiosis and possible preventioigi of 
the diseases. 

The activities of the Branch are carried out through a number 
of operational methods, oste of which is an extensive program of 
cooperative research investigations (direct field studies) in which 
staff meffibers work directly with physicians, clinical isivestigators 
and scientists in medical schools, teaching centers, universities, 
hospitals and cancer centers throughout the nation. Some sixty °five 
such collaborative research studies were under way iss 1959. A secoatid 
method of operation involves field studies in which the entire iii^vesti^ 
gative group is composed of staff members directly responsible to the 
Bramch. Tl^e Coffman Research Laboratory in Hagerstowra, Marylaaad, is 
an example of this method. Still as^ther means of operation is through 
several extraisural gramt programs which are comoiented upoa later in 
this report. The last means of operatio!£i involves the use of cosstracts 
to procure research. This last^mentioned method has come into itticreased 
usage during the past year particularly in the diagnostic research a^d 
development program. 

One of the sigMificant accomplishments of the Branch during 
the year has been the establishment of a broad program in diagnostic 
research. This :ps@gram, based on the counsel of several advisory groups 
representing a large nuoiber of clinical and scientific disciplines;, 
undertakes to increase research efforts in the fields of clinical 
cnsymology, clinical immunology, clinical chemistry, clinical pathology, 
endocrinology, steroid chemistry, fluorescent microscopy, tissue culture 
and clinical cytology as applied to the problems of diagnosis of human 
cancer. Eleven contracts totaling approximately $471,000 were negotiated 
and awarded is this program during 1959. A number of additional contracts 
were brought through the preliminary stages of negotiation. During the 
same period the Branch conducted some 17 investigations related to this 
diagnostic development area (NCI Project Nos. NClo85, 105, 108, 109, 
110, 144, 145, 146, 147, 149, 152, 160, 163, 165, 166, 167, 168). Nine 
publications by staff roenobers associated with these projects were 
published during the year. Their titles are noted on the attached 
Institute project reports. Closely allied to this area are the investi'^ 
gations conducted by the Branch in the field of exfoliative cytology. 
Sixteen such studies have been in operation during the year (NCI Project 
Nos. NCI°81, 83, 84, 86, 87, 88, 89, 90, 91, 94, 95, 96, 103, 104, 106, 
107) . By and large these have been conducted as cooperative research 
ii^vestigatios^ in conjunction with medical schools and universities o 



74 



- 2 - 

Duxii^g the year pxelim±tmty steps ^?ere takesi eo pluase out Che data<° 
c©llec£i®g ee^nsRts ©f fehe exfoliativa cytology studies of cervical 
camcer issi five projects (Columbus, Meisphis, Madison, Louievilla, 
Sas!i Diego) . At the same titiie efforts i&a\m been made to increase 
research invest igatioms of exfoliative cytology of other casicer sites: 
colon and large bowelj luag, gastric (Projects Nos. NC1>=83, 86, 87, 
88, 103, 104, 107)9 aad urissary tract. Prelimiaary arrangements ware 
made for studies of the last^mentioned site just at the close of the 
year. Some 25 publications astd preses&tatioms by staff mecibers and 
uaivarsity faculty members associated with the cytology projects ware 
published and presented during the year. Their titles are noted oa the 
attached project reports. Oste of the most significant studies started 
during the year was the clinical trial with the Cytoanalyzer in ^ich 
it is anticipated that 10,000 to 15,000 specimes&s will be processed 
through the machine and the results analyzed. 

Ai^iOther major area ®f Branch activities is in the field of 
eaviroranental casacer studies. Seventeen studies in^rolving possible 
environmental factors aad their relationship to cancer vere conducted 
duriag the year (NCI Project lifos. KCI»lll, 112, 113, 116, 117, 119, 
120, 121, 125, 126, 127, 128, 129, 130, 131) « Direction for seven ©f 
these studies is centered in Hagerstosa, Marylaad; six in Sil\f«r Spriag, 
Marylaad, three isi Boston, Massachusetts, asid ©as in Salt Lake City, 
Utah. Most ©f the®« investigation® are by mature loag-rassga in 
character sm.d therefore publication of results has been limitedc 
However tf?o publications by staff meisbers are noted on the attached 
project reports. 

Some 18 general aiad epidemiology field studies havs beaa c©ffl» 
ducted ©r concluded during the year (NCI Project btos. NCI»92, 139, 140, 
141, 142, 148, 150, 153, 154, 155, 157, 159, 161, 164, 170, 171, 172, 
173) o Most of these studies are epidemi® logical and statistical in 
nature and must await the accumulation of sufficient data fer aaalysls, 
Dae of the most significant im this group is the analysis ©f the data 
compiled in the Men?>his cervical cytology investigation ^-hich should 
be ready for publication in early 1960. Ten publications prepared by 
staff associated with these studies are noted on the project reports. 

Asaotfeer area of activities of the Beassch is coacerssed with cancer 
nursing (NCI Project N©s. KCI-182, 183, 184, 188, 193, 194, 195, 196, 
197)0 One of the major xmdertakings during the year has been to start 
the preparation of a Mos^graph oa Radiation for Nurses; asother study 
started in 1959 is concerned with the problems faced by patients uader- 
going cancer therapy; other studies are concerned td.th aurse-cancer 
patient relations and casscer teaching for nurses. One publication is 
ffioted on the attached project reports. 



75 



- 3 - 
Gran£ Programs 

The Branch coafeimued isa 1959 £o operate tha followiisg grant 
programs: field investigation grants, ussdergraduate cancer training 
grants, cancer teaching project gragsts for schools of snursinij and 
public health, clis&ical traiase® ships, cytology techaician traisseeships 
and radium loans. 

Requests for support under the field laves tigat ion grants program 
increased both iM number and total dollar volume. Amossig the approved 
requests there was a marked increase in the support of invefitigatlosiis 
in the areas of diagiitostlc, applied, clinical and epidemiological 
research projects. Of major sigstificance was the establishments by 
staff members utilising the grant mechanism, of a National Cooperative 
Study of the Etiology of Leukemia in 12 to 15 locations throughout the 
country . 

Ira addition the staff was resposasible for stimulation of grasjE 
requests in the field of diagnostic research and developmasit, some 
15 of which totaling approximately $300,000 were approved and supported 
by this Branch, as well as a sizable nudbsr of a more basic nature 
which were supported by th« Research Grassts Branch. 

Although there were recommendations by the National Advisory 
Caacer Coustcll and other advisory groups to increase the levels ©f 
support of the laadergraduate training grant program, it was confcinusd 
at the same support level as in previous years. This program continues 
to be an extremely worthwhile endeavor, a fact which is attested to by 
results of surveys made of medical schools by special task force teams 
of the National I&istitutes of Health, as well as results of the cancer 
knowledge testing program, support of which was discontinued during 
the last year. 

Under the cancer teaching projects activity four awards were made, 
two to schools of nursing and two to schools of public health to 
strengthen cancer teaching for their students. In regard to grants to 
schools of nursing for improvement of cancer teaching the National 
Advisory Cancer Council reconananded that this program be extended and 
that funds to support an expanded program be secured. However this 
was not accomplished during the year. 

The clinical traineeship program which supports graduate physicians 
in special training in the diagnosis and treatment of cancer was con- 
tinued as in previous years. Some consideration has been gives to 
changing this program from a direct trainee support program to an iffidirect 
training grant award program. 

Tfee cytology technician traineeship program was expanded during 
the year so that practically double the mumber of individuals, approxi- 
mately 95, were supported in 1959 as contrasted to 1958. 



76 



o 4 



Ussder the radium l©aa prograia practically all ©f the available, 
usable radium has beea assigned t© radiologists ia fifty iasfeieutioaa 
throughout the couatry. Steps have been take© to tnoderaize this 
program by coKversiois of eome of the uaits of radium to more usable 
sizes and dosages- This cosiversion will be proceeding at a rate ^ich 
will see it accomplished over the raext t^so years. 

Details of the total activities of the Branch are more specifically 
outlined ira the attached individual project reports. 



77 



k 



SCIE1ST.IFIC PAPERS PUBLISHED WKmQ 1959 
PIBB-KCI 



Blanco Be Del Casj^y Maris So easd Edelsoa^ Stuas't Hoj A gisiple nje-^od for .■ 
prepariag Hsicro©C5opie slide® of sgasamaliaa cells gro^m In vitro; Tke cottoa 
tmah Esaear tec&niqueo Mc Jo Cliao Pat&o, 32;^«i*C^^ Oeto 1959o 

Calabxegij, Psul & Mej^ier^ Ovid 0<, s f&lycsrKfeesBia vei^: I-Cliaical aaid laboratory 
manifestations, itonals of Intersil Med. 50:ll82''1202, May 1959. 

Calabreei, Paul & Meyer, Ovid 0. : Bolycythesnia vera: II-Cour£e and 'BieT3.-py. 
Annals of Internal Med. 50:1203-1216, May 1959. 

Calabreei, Paul, Edwards iferl A. & ScMlling, Robert F. j Pluoarescent anti- 
globulin studies in leukopenic and related disorders. J. Clin. Invest. 
38:2091-2100, Hov. 1959. 

Carroll, B. E. and Ingraham, S. C: A recoM and reporting system for field 
research units. Pub. Hltb. Rpts. 7^:885-890, Oct. 1959- 

Dom, Harold F. : Tobacco consumption and mortality from cancer and o13ier 
diseases.. Pubo Hltaio Rpts, 7^:581-593 (July) 1959- 

Dunn, John E. and Buell, Hiilip: Association of cervical cancer Hith cireusision 
of sexual partner. JKCI 22:7^9-76^* April 1959- 

Dunn, John E. , Slate, lliomas A. , Merritt, John y. , end Martin, Purvis L. : 
FiadlngB frosa cytologieal exaiaination of 33^750 ^'OHien one or more times for 
uterine cancer. JNCI 23:507-52?, Sept. 1959. 

Geiser, Patricia: Bibliograpiby on cancer for nurses. Pub. Hlth« Ber. Publ.Ko. 68?. 
Pub, Hlth. Bibliograji]^ Series No. 26. U. S. Govt. Print. Office, WaeMngton, D. C. 
1959. 

Grinspoon, L. and Bunn, J. E. : A study of the frequency of achloAydria asong 
Japanese in Los Angeles. JICI 22:617-631, March 1959. 

Joslia, Eo P., Losabard, H. L., Barrows, R, S. and Manning, M. D. : Diabetes and 
cancer. New Eag. J. of Med. 260-1^86-488, March 1959-' 

Eahl, Ruth & Brass, Mattie: Cancer Kursing Consultation. Nureing Outloo;^ 
7: , March 1959. 



78 



Lam'eaee^f P. A, & Chen v7. Y. ; A project for ©tiid;^ag the geographic distribu- 
tioa of cancer {yitMB & single county as s-elated to envii'^aiaeatal facbox^e, 
Ajra k9'MB^6fh, May 1959. 

Lundin, Fraak E, and Ross, Sister Hilasy: llrer disfUBction in leprosy. 
Intemat. J, Leprosy SfjUS-J^T, 1959. 

Milaore, Bsnno K. : Book jreview of "Variables related to Jjuuian breast esncer" 
by V, ELviag Anderson, Hferold 0. Goodiaaa asad Sheldon C. Eeed, the Univ. of 
Mimesota "PreBBf Mianeapolis, 1958. M. J. Bib, lealtli ^^9: 1271- 1272, Sept. 
1959. 

Pii-'tersoa, jRoealie I.: Overview of rehabilitation aureiag: Is it ne^ or is 
it 0:1 d? Military Medicine 12ij-:28^-287, April 1959. 

Pruitt, J. C.J Hilberg, A. W.j and KMser, S, P. j Isolation and identification 
of ceacer celle in perijiieral blood. J. Abdom. Siirg. , May 1959- 

Pruitt, John C, Eilberg, Albert W., Kaiser, Raymond F., Izigrahmi, Saausl C, 
Smith, Sai«2i J. and Willoughby, Maiy B.: Spray teebniqiae for preparation of 
cytologic epecimenB for automatic scanning machines. JKCI 22:1105-1117* June 
1959« 

Sievers, M» L, and Galls^aer, N. I,: Interrelationship of acid, pepsin, and 
intrinsic factor production. An indirect gastric secretory study. Gastroenter- 
o].ogy 57:182-187, Aug. 1959- 

Sievers, M, L. and Calabi^si, P.: Gastric pepsin secretion and AEO blood groups 
in polycythemia vera. Am. J, Digest. Dis, 4:515-521, July 1959- 

Sievers, Mo L. : Hereditaiy aspects of gastilc secretory function. Eaee and 
AK) blood groups in relationship to acid and pe|®in production. Am. J. Med. 
27.°2U6-255, Aug. 1959. 

Taylor, Re S,, Carroll, B. E. and Lloyd, J. ¥,j Mortality anong -aomen in 3 
Catholic religious orders -^th epecisl rsfereaea to csicer. Cancer 12;1207- 
1225, Bee. 1959. 

Vender, John B. : Optical density laeasurements of leukemia cells. Cancer 
12:2^-23^^, March-April 1959» 

Voider, John Bo: TSie agsoeiation of diabetes laellltuB and carcinoma of the 
endometrium. Aa. Jo of Obetet & Gynec, 77:243-2'4-5, Feb. 1959- 

VogLer, WLlllaa Bo & Bavell, Baljii Waldo: A clinical evaluation of -aersaography 
and heptyl aldehyde in breast cancer detection. J. din. Inves. 19:207-2095 
Feb. 1959. 



79 



PAPERS (MmJSIW MB ACCEFCED FOB PUBLICATIOH 1959 

PIDB-NCI 

Archer, Victor E. and Carroll, Benjamin E. : Life shortening and tiAiEor production 
by SR-90. An alternate presentation of the data given by Finkel in "Mic©, men 
and Fallout". Approved for Science. 

Archer, Victor and Luell, Edith: Effect of iseleniijm siilfide suspension on 
hair roots. Approved for J, Invest. Dermato 

Archer, Victor E. and Simpson, C Lenore: Relationship of radiation and neoplasia 
in man. Approved for Am. J. Roentgenol. Rad. Ifeerapy & Nuclear Med. 

Courtney, Williem B. ; Eilberg, Albert W. ; Ingr^em, Semuel C. ; Kaiser^ Rayiaond F. ; 
Pruitt, John C. ; Bouser, Mary M. : Field trial of tha cytoanalyaer: II86 specimeng 
analyzed. Approved for JKCI. 

Courtney, William Bo ; Eilberg, Albert W. ; Ingrahaa, Samuel C. j Kaiser, R^?Koad F. ; 
Pruitt, John C, Bouser, Mazy M. : Field trial of t&e Cytoanaiyzer: il86 
Bpecimeffig analyzed. Presentation Inter. Soe. Cytol. Cotmcil and publication in 
transactions of IoS,CoC, (Nov. 19-21, 1959). 

Dunn, John E. j, Jr. , Mo D. : "Hie epidemiology of cervical cancer as revealed by 
cytolo^c For presentation at laeetings of latei^iat. College Surg., CMea^., lllo 
9 ''16 '59 @3id publication in proceedings. 

Dunn, Marvin R. • The incidence of carcinojaa arising in cervical stuaps. For 
presentation at sseetings Inter. Hoc. Cytol. Ccua,cil and publication in tssngaetior 
(l?0Vc 1921, 1959) » 

Hoaglin, LeMar W. , Linden, George, Breslov, Lester and Bunn, John E, , Jr. : TtiB 
use of a sailed questionnaii^ for epidemiological study. Submitted to Pub, Bith. 
Rep, 

Levin, Mu L., Haenszel, ¥,, Carroll, B. E., Geiiiardt^ P. R. lanSy, V. E. and 
lagrabaa, S, C.s Cemcer incidence in urban and rural areae of H. Y. State, 
Submitted to .rwci. 

Peacock, Andrew C, and Williams, G, Z. ; Biological reactivity >^f substituted 
2,3,5 tsl|3ienyl tetrazoliusas. Presented at Fed, Proc, of -the M. Soe. for 
Erper. Pat^, and publication in ts-aaeactions. 

PeiiSergraet, William J,, Milmore, Besno K, and Mareiis, Samuel C: liiyroid cancer 
and thyro toxicosis in iie United States. Their relation to endesnic goiter, ISev 
Approved for Hew Eaglceid J. of Med. 

Pruitt, John G.; Carpenter, Bsarxy M, : Eilberg, A, ¥.| and Morehead, Bsbert P.: 
Quantitative isolation and identification of malignant cells froa circulating 
blood and body fluids. Presentation at meetings of Soc. of Sxper, Biology and 
Med,, Charleston, S. C, Oct, 30, 1959, and publication in transactions. 

Staffbrd, Marvin W, : An improved method of labeling frosted-end microslideB 
used in exfoliative eytolo^. Tech. Bull . Amer. J. Clin. Pathology. 



w 



U abet, BayaoM J. , Cajseron, A„ Bruce and Ktaoejasdiild, Earvey E. : Value of the 
Eillipore teehnie in the cyto3.oglcal diagnosis of earclEoma of the colon. PrsB. 
at meetings of Intar. Soe. Cytol. Council and publication in Soc. tresisactionB. 
(Nov. 19-21, 1959). 

Vender, John B. : daesifi cation of leukeaias by ultra violet absorption laeasure- 
aente. Approved for publication in Cazicer. 

Vander, Jbbn B, 85 Jobnson, Horton A. : Chronic lyiaj^atic leukeaia and sultipLe 
Bayeloma in the same patient. Approved for Mevo J. Med. 

Willl©a, G. Zo & ^acocfe, A. C. : Ultraviolet studies of tetrazolium reduction 
in living cells c A^pcroved for Exper. Cell Reso 



81 



Serial Nc HCI~8l 



PHS-WIH i„ Field Investigations and 

Individioal Project Report Demonstrations Branch 

Calendar Year 1959 2« Cytology Section and Diagnostic 

De-velopnen''fe Program 



Silver Spring, Maryland 



Part A, 



Project Title: Supervision and administration of research projects 
investigating exfoliative cytologic methods aaid 
techniques as applied to various human body sites 
and of diagnostic development program 

Principal Investigators: nr» S. C. Ingrahajn, II, Dro Ao Wo Hilberg 

Miss Mo Mo Bouser, Miss Anne Baranovsky 

Other Investigators: MrSo Kathleen Co Fant 

Cooperating Units: Office of the Chief, Field Investigations 

and Demonstrations Branch 

Man Years (Calendar Year 1959): 



Total : 


5^ 


Professional: 


4 


Other : 


1% 



Project Description: 

The activities of the Cytology Section, FIDB, consist of many 
varied administrative and scientific functions in' relation to the 
several. Field Reseaxch Projects concerned with exfoliative cytology 
of huiran cancer o Among the activities are included: 

1} Planning, for and with the Chief, FIBB, the policies of 
the Cytology Program of the Branch and guiding the program and 
developing the projects in directions consistent with these policies, 

2) Plsaining new research projects and esta-blishing the 
projects in university medical schools or similar scientific groups „ 

3) Recmitment and orientation of professional and non- 
professional employees engaged in the research studies- 

k) Consultation with professional staff of the field projects o 
This includes both non-Public Health Service professionals and 
Public Health Service professionals o 

5) Coordinating activities of all field research projects 
with each other autid, as requested, with outside professional 
groups such as university medicaJL staffs and medical, societies. 



i2 



COIWimJATIOW SHEET - 1 Serial No, NCI-81 



Project Description : (conto) 

6) Rendering assistance to all field research projects, upon 
request, -sdaen problems arise relating to scientific or adminis- 
trative activities concerned with exfoliative cytology as applied 
in the project « 

7) ifeeping project personnel informed concerning recent 
publications and information about exfoliative cytology and 
related scientific topics. 

8) Participating as requested in cancer research activities 
of national, state, or local medicaJL and scientific organizations 
as active members of coordinating committees, advi&ory committees, 
etCo Also talcing active part in panel discussions and formal 
presentations as invited by such groups « 



Peirt Bo included Yes /X/" No [J 

^ 83 



i 



PHS NIH Serial No« NCI-8I 

Individual Project Report 
Calendar Year 1959 



Part B . Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Gilliam, A= Go, Kaiser, R. F.,, and Ingraham, 2d, S= C.: The 
Role of the Federal Government in Cytologic Investigation o Acta 
Union Internationale Contra le Cancer, Vol. XIV, Wo. k, pp. ^1-33-437, 
1958. 

Kaiser, Ro F« and Hilberg, A» W. : Changing Goals in Cytology 
Resestrcho Bxilletin of the College of American Pathologists, 
Volo XIII, ppo U9-5O, April 1959. 

Pruitt, J. C, Hilberg, A. Wo, Kaiser, Ro F., Ingraham, S, Co, II, 
Smith, So Jo and Willoughby, M<, Bo : Spray Technique for Preparation 
of Cytologic Specimens for Automatic Scanning Machines » Joiurnal of 
the National Cancer Institute, Volo 22, ppo 1105-111?, June 1959^ 

Pruitt, J. Co, Hilberg, A. Wo, and Kaiaer, Ro Fo: Isolation 
and Identification of Cancer Cells in Peripheral Bloodo Journal of 
Abdominal Surgery, pp. 37-40, May 1959o 

Co^lrtney, Wo Bo, Hilberg, Ao Wo, Ingraham, So Co, II, Kaiser, Ro T 
Pruitt, Jo Co and Bouser, Mo Mo: Field Trial of the Cytoanalyzer : 
1186 Specimens Analyzed. Journal of the National. Cancer Institute 
(in press) o 

Carroll, Bo Eo and Ingraham, II, S. Co: A Record and Reporting 
System for Field Research UnitSo Public Health Reports, Vol. "jk, 
ppo 885-890, October 1959 » 

Pruitt, J. Co, Carpenter, H. M., Hilberg, A. W„ and Morahead, R. P 
Quantitative Isolation and Identification of Malignant Cells from 
Circulating Blood and Body Fluids. Presented at meeting of the Society 
of Experimental Biology and Medicine at Charleston, So C on October 30, 
1959^ niay be published in the Journal of Experiinental Biology and 
Medicine , 

Kaiser, R. F., Bouser, Mo M., Ingraham, II, S. C, Hilberg, A. \-L 
ajid Baranovsky, A.: A Preliminary Summary of Results of Cytologic 
Examinations on 600,000 Women. .Presen^ted at Inter-Society Cytology 
Coimcil Meeting at Detroit, Michigan, Norember 19-20, 1959 and appx-oved 
for publication in Public Health Reports o 



84 



CONTINUATION SHEET - 1 Part B. Serial No. HCI-81 



Publications other than abstracts from thit> project: (Conto) 



Dr. IngraJham spoke to the Orientation Group of the Cancer 
Control Program, PES, at the GSA Regional Office Building on 
July 8, 1959 on the cancer diagnostic development program of 
the PIDBo 

Dro Hilberg spoke on the subject "Cajicer Cells in the 
Peripheral Blood Stream" at a Cancer Symposium at the Alumni 
Association of the Jersey City Medical Center, Jersey City, No J« 
on November h, 1959 « 

Miss Bouser took part in a working conference for Occupa- 
tional Health Nursing Consultants from State Health Departments, 
January 7-9, 1959 at Cincinnati, OhiOo 

Exhibit: "Studies of Cancer Cells in Circulating Blood" was 
displayed at Public Health Service Meeting in New Orleans, La., 
in March 1959; at the North Carolina State Medical Association 
Meeting in Asheville, N, Co in April 1959; and at the Clinical 
Meeting of the American Medical Association in Atlantic City, 
No Jo in June 1959". 



Honors ajid Avards relating to this project: 

The exhibit listed above won the Ceirfeificate of Merit in 
the Preventive Disease Section at the American Medical Association 
meeting in Atlantic City in June 1959 » 



85 



Individual Project Report 
Calendar Year 1959 



i« Field Investigations and 
Demonstrations Branch 
2„ Cytology Section 
3« Columbus, Ohio 



Part A. 



Project Titles Early detection of carcinoma of the colon by 
cyto logical and chemical techniques. 

Principal Investigators: Dr, Emmerich von Haam, Dr, Robert Zollinger, 

Dr, Harvey E, Knoernschild, Dr. Raymond Thabei: 
Dr, Walter Frajola 



Other Investigators: 



Dr, R, F„ Kaiser, Dr, S„ C. Ingraham, II. 
Dr„ A„ Wo Hilberg 



Cooperating Units; The Ohio State University Health Center 
The Columbus Cancer Clinic 

Man Years (Calendar Year 1959): 
Total : 20 
Professional; 2 
Other s 18 

Project Description: 

Objecti ves: The purpose of this project is to develop a chemical 
and/or cytological test for carcinoma of the colon and rectum. This 
program, if successful, could make practicable earlier detection o£ 
these cancers and help decrease the morbidity and mortality from 
malignancy of the colon and rectum in the general population. 

Methods Employed: Patients are examined clinically and specimens 
obtained via proctoscopy ^ sigmoidoscopy and examination of enema 
returns for laboratory testing and evaluationo Biochemical and 
histologic tests are being devised and tried to identify changes presen'i 
in patients with carcinoma of the bowel. Cytology procedures are being 
devised to apply this technique to diagnosis of bowel cancer. 



The project is divided into four sections 
clinical research, chemistry and cytology,, 



administration^, 



The administrative section compiles and files records and 
laboratory data and performs other administrative functions 
necessary to the project. 



86 



COKTINUATION SHEET - 1 Serial Mo. KGI-83 



Methods_Eggilg;ged g ( cont , ) 

The clinical progras performs signioidoscopies routinely on 
patientis on the Surgical Service of the University Health Center and 
on patients in the Columbus Cancer GliniCo Selected additional 
patients in the Health Center Medical Service are also sigmoidoscopedo 
Specimens are obtained for the chemistry and cytologic activities and 
a con^lete record of sigmoidoscopy findings is maintainedo Special 
patients are singled out for eKamination of enema returns cytologically„ 
The number of sigmoidoscopies performed monthly averages about 400 » 

The chemistry program is investigating the possibility and 
practicability of using fecal blood detection as a mass screening test 
for carcinoma of the colono Such tests are routinely performed on 
patients entering the Ohio State University Hospital on the Medical 
and Surgical Services o 

The cytology program is being furnished with mucosal scrapings 
from sigmoidoscopies and is examining surgical specimens to discover 
whether a cytologic examination may be helpful in the detection of 
pre^malignant or early malignant lesions of the colon and rectum„ 

The use of the Millipore filter in obtaining cytological material 
from enema re' urns has been used clinically since January, 1959 » 
Patients are selected for this study on the basis of significant bo'wel 
symptoms. Am average of 10 patients are studied weekly by this method- 

Patient Materials Patient material is derived from the inpatient 
and outpatient population of the Medical and Surgical Services of the 
Ohio State University Health Center and from the patients of the 
Columbus Cancer Clinic „ 



1) In 9982 sigmoidoscopies to date, data is coaaplete in 8622 
patients. 

a) The incidence of colonic polyps has been 6.57eo (564 polyp 

b) Twelve percent of biopsied adenomatous polyps have been 
malignant polyps, or an incidence of 0»42% of all patients 
examined o (36 cases) 

c) Of all patients examined, 125 (lo457o) have had carcinoiiia 
of the colon, 

2) Of the 8622 patients 7521 were examined with sigmoidoscopy 
cytology (87.51) o 

a) Of the 100 cases of carcinoma of the colon examined by 
this method, 51% had malignant cytology, and 16% had atypical 
cytology o 

b) Of the 22 cases of malignant polyps 6 (277o) had malig- 
nant cytology, and 13% had atypical cytology„ 



87 



CONTINUATION SHEET = 2 Serial No, NC1-S3 



Major Findings g ( cont = ) 

c) The incidence of malignasjt cytology in the beBign 
polyps was 5.37»» An additional 197. had atypical cytologyo 

d) Of 6146 normal colons only 0,16% had malignant cytology 
and 2,67o with atypical cells, 

3) Of the 8622 patients in the completed group. 6431 have had 
their feces examined foe blood (757o). 

a) Of the known carcinomas of the colon in this group (89) 
60% were positive for fecal blood, 

b) Of the known malignant polyps in this group, 307, were 
positive, 

c) Of the known benign polyps in this group ^ 13% have had 
positive blood, 

d) Of the cases in this group v^ith normal colon, 10% have 
had positive fecal blood, 

4) Early results are available from the Millipore filtratiosi c f 
enema returns, A total of 441 cases have been completed„ 

a) Of 31 cases of malignancy of the colon^ malignasE cells 
have been recovered in 80%, 

b) Of 4 cases of malignant polyp s^ 50% have had malignant 
Millipore cytology, 

c) Of the 252 normal colons, there have been no false 
positives, (One has occurred since the date of this report).-. 

5) By combining the two cy to logical methods currently eH^loyed, 
88% of the carcinomas of the colon (25 cases) studied by both mefchcds 
have been detected cytologically, 

6) In comparing the two cytological techniques ^ the Millipore 
filter technique appears to be a more reliable method for the 
detection of carcinoma of the colon above the 25 cm, sigiaoidoscopic 
range. In contrast, the direct smear method is more reliable in 
lesions lying within range of the sigmoidoscope , but only if seen 
Airing sigmoidoscopy, 

Slgaif l eaacfc to the ?ro;p;ram of the Institute; This study by 
developing methods for detecting and diagnosing cancer of the large 
bowel in its early stages, will, if successful, offer a means whereby 
this type of cancer might be mora readily controlled in the general 
population, 

£8 



CONTINUATION SHEET 



NCI°83 



Proposed Course of the F goigct s Data collection, the study of 
methods, inqproved collection of statistical methods will probably 
continue for approximately an additional 1% years to 2 years. By 
this time it is anticipated that statistically valid numbers will 
have been accumulated. 



Part Bo included 



Yes /X / 



No 



69 



PES'-NIH Serial No„ KGI_"°83 

Individual Project Report 
Calendar Year 1959 



Part Bo Honors, Awards, and Publications 

Publications other than abstracts from this projects 

The paper "Recovery of malignant cells from enema returns from 
carcinoiaa of the colon" by Drs. Ao Bo Cameron aM Ro Jo Thabet 
was presented before the Ai&erican College of Sux'geons, September 
1959 o 

The paper "Sigmoidoscopy as part of routine cancer clinic 
examinations with correlated fecal cfeejEistJry and colon cytologic 
studies" was presented by DrSo Ac Bo Caiaeron and Eo Jo Thabet 

before the Ohio Chapter, American College of Surgeons, Septj 1959 •. 

The paper "The evaluation of chemical and cytologic leethods of 
detection of laalignaacy of the colon and rectum" was read by title 
by DrSo Ac Bo Cameron, Ro Jo Thabetf G. Yo SMnowara, EoVOis Kaam, 
and R„ Mo Zollinger at the American Gastroenterological AssociatioB 
in May 1959, 

The paper "Value of the Millipore technique in tfee cytological 
diagnosis of carcinoma of the colon" was presented by DrSo Ro Jc Thabs 
Ao Bo Cameron, and H„ E„ Knoernschild to the Inter^Society Cytology 
Council in Detroit, November 21 , 1959. 

"Sigsaoidoscopys A useful tool in the cancer problesa" by Bx. H„ E. 
Knoernschild was published in the Columbus Aeadeiay of Medicine Bulletiv 
Vol. 25, po 29, November 1959. 

A scientific esshibit "The deE3ons«r.ration of saalignatit cells Ik 
lesions of the colon" was pr'epared and presented by BrSo Ro Jo Xhabet. 
A. Bo Cameron^ H„ Eo Knoernschild, and B, Macfsrlane for the Inter- 
Society Cytology Council in Detroit from Kovember 19-21 , 1959o 



Honors and Awards relating to this projects 
None 



90 



I 



ms-mE 

Individual Project Report 
Calendar Year 1959 



Serial No, NCI --84 

1. Field Investigations and 
Demonstirations Branch 
2c Cytology Section 
3o Columbus, Ohio 



Part A, 



Projec£ Titles A study in a general populatiom of cervical cancer 
using vagiraal and cervical cytology for p'^~litninaj?y 
case selection - Columbus » Ohio 

Principal Investigatorss BSo Emmerich von Haanis Dr. John Ullesys, 

Drn Edward Miller 



Othes" Investigatorss 



BTo S, Co Ingrahams II^ Dr, R, Fo Kaiser, 
Bw. A. W= Hilberg, Miss M» Mo Boueeifs 
Miss Anne Baranovskyj, Mrs, Doris Fisk, 
Miss Margaret J. Caiseron 



Cooperating Unitss 



Ohio State University Medical School 



Man Yeasss (Calendar Year 1959); 
Total I 31 
Professional; 3 
Other 5 28 

Project Description; 

Object i ves ; 1) the evaluation of the vaginal aspirate technique 
for reliability of the case identification method for cancer of the 
uterus in the adult female population of Columbus ^ Ohio^ 

2) the relative comparison of the vaginal aspirate and the cervical 
scraping methods » 

3) compilation of information regarding various epidemiological factor 
involved in carcinoma of the cervix for evaluation and correlation 
with the different types of uterine carcinomas found. 

Each type of cancer detected will be evaluated as to the typa of 
treatment received and follow=up studies evaluating survival wii ' 



type of treatment will be obtained. 

Cytologic material is to be supplied for the clinical evaluation 
trial of the Cytoanalyzer in Hagerstown, I4aryland„ Their data will be 
correlated with results obtained by the standard staioissg and screening 
techniques . 



5^1 



CONTINUATION SHEET » I Serial KOo KCI={ 



Methods Eng)loyed; The adult female population of Greater ColuiabuSs 
Ohio 5 is being used as a sou^'ce of patient material for determining 
the effectiveness of the vaginal cytologic smear method and other appro 
diagnostic tests and procedures for determining presence of cervical 
cancer o 

1) By means of the vaginal smear method and other medically 
accepted cancer diagnostic tests and procedures an investigation is 
being made of the adult female population in Greater Columbus j, Ohioj 
for occurrence of uterine cancer „ 

2) Cancer cases detected are evaluated and treated appropri- 
ately by usually accepted medical j, radiological, and surgical methods „ 

3) Appropriate clinical and medical°statistical records are 
being kept to provide evaluation of uterine cancer occurrences adequate 
patient=follow=up, incidence of false negative and false positive case- 
identifying results J and efficiency of the several case^identification 
and diagnostic methods « 

4) This study consists of two interrelated but physically dis- 
tinct operations; 

a) Relative evaluation of case-=identification and diagnostic 
techniques - in this portion of the study essphasis is placed upoa 
the thoroughness with which a large group of patients is esasninedo 
Here the methods of exfoliative cytology ei^loyed are the taking 
of a vaginal smear according to a standardized techniques plus the 
taking of cervical scrapings after exposure of the cervix by 

If, speculum. Inspection and description of the cerviK also is record 
by the examining physician or medical student. The several labora 
tory specimens are examined and evaluated separately and the find- 
ing of the several methods collated, correlated, cossspared, s^d 
analyzed, 

b) Ca8e= identification for relative occurrence of uterins 
csmcer in age=specific and other specific population characterl; : 
in this portion of the study enjphasis is placed upon offering 
vaginal aspiration exfoliative cytology to large population sag" 
meats on a periodic recall basis j specific diagnosis by accepted 
clinical and laboratory methods;, and appropriate therapy of all 
patients found to have uterine cancer. Patient records are being 
analyzed for incidence, prevalence » and other pertinent factors 
concerning uterine cervical cancers its detection and treatment 
in an adult female population. 

Patient M aterials Patient material is the adult, female population 
of Greater Columbus, OhiOs including clinic and hospital patients j 
indigent patients » patients of private physicians, industrial workess, 
and the presumably healthy female population of the area. 



92 



30riTINUATIOK SI-IEET - 2 Serial Hoo K CI _-:8,4 

Major, F iadlags s 

A total of 213 1,680 slides have been received to date 
(KovembeE' 20 » 1959) from three major sources: 

1) The Cancer Survey Project Cliaic has obtained 13a442 slides 
(6%) , This figure also includes specimens obtained in the State Mental 
Hospital and from several separate industrial and commercial organiya 
tions where special arrangements were made to offer the cytology sesewsj 
ing test to en^loyees, 

2) The majority of slides (155 ^040 or 73%) has come from privafce 
physicians in Franklin County^. Included in this group are slides from 
five clinics which have been established in all the larger Columbus 
hospitals and which function similarly to the Cancer Survey Project 
Clinic. Over half of all active physicians in Franklin County have 
participated in the project. Most of the others have no gynecological 
practice. Proven positive cases have been detected in the offices of 
over 120 physicians. 

3) The remainder of the slides (45,198 or 217. has come from 
various out-patient clinics of the Ohio State University Health Center. 
and from the Cancer Clinic » 

The total number of cases to date is 100s859o The difference 
between this figure and the number of slides received represents cases 
where both vaginal aspirates and cervical scrapings are received on tfee 
same patients and on cases where repeat » first recall, and second reca} > 
examinations have been donej 35,457 cases have had two or more eKaraina 
tions at annual intervals,, Third examinations have been received on 
I0s,615 patients and 1,051 have had four or more examinstionsc 

A summary of the results of the interpretation on initial r.ytol'.:x_ 
screening first screening to date is as follows? 

Negative 98,23% 

Unsatisfactory 0«7I% 

^ Suspicious 1„00% 

^ J . A Tota l jjf 463 cases of malignancy has been detected to date. 

The recall program is progressing very satisfactorilyo Based on 
the number of cases in our files one year previously^ our first recall 
follow-up is 46%- 

All record cards have been microfilmed and sent to the statistics 
unit of FIDB, Preliminary analysis of these cases is nov7 in progrssi, 



93 



CONTINUATION SHEET - 3 Sex: 



Significance to the pgoggam of the Insti tutes The results o 
study will be added to the informatioa being obtained in othe'" c^ 
logical studies o Consolidated data are expected to furnish insij 
into pathogenesis of uterine cervical cancer with prevalence and 
incidence data in tne adult remaie populationo 

Proposed Cou rse of the grojeet s The study is to ba concluded by 
July 1960 following a four year programo 



Part B included Yes / / No /X / 



94 



PHS-NIH 

Individueil Project Report 

Calendar Year 1959 



Serial No. NCl-85 

Ic Field Investigations and 
Demonstrations Branch 
2. Diagnostic Development Pr-cgTE 
3» Winston- Salem, Worth C*r',lir . 



Part A o 

Project Title: Peripheral Blood Project - Winston-Salem, N, C 

Principal Investigator: Dr. John Co Pruitt 

Other Investigators: Dr^ Robert P, Morehead, Dr« Henry Fo Mengoli, 
Dr. Ao Wo Hilherg 



Cooperating Units: 



BoTflnan-Gray School of Medicine and 
North Carolina Baptist Hospital 



Msji Years (calendar year 1959) ; 



Total : 




Professional: 


Other : 


3 



Project Description: 

This is a research project for the purpose of examining human 
blood specimens for circulating malignant cells and investigating 
immunologic techniques of destroying those cells or preventing 
their growth « An attempt is being made to answer the following 
questions: 

1) What percentage of patients witJi cancer have cancer cells 
circulating in the peripheral blood? 

2) What is the relationship of malignant cells in the periph- 
eral blood and prognosis? 

3) What percentage of patients have malignant cells in ■i'ae 
veins draining their tumor sites at the time of surgery? 

k) Does manipulation of a t\amor at the time of surgery cavise 
a spraying of malignant cells into the blood stream? 

5) Do any types of cancer sited malignant cells into the blood 
early enou^ and with enough regularity to make the test of diagnostic 
value? 

6) Are cancer cells shed into the blood in a continuous stream 
or intermittently? 

7) Is there any way to determine if the cancer cells identified 
in the peripheral blood are viable? Is this related to prognosis? 



95 



CONTimATION SHEET - 1 Serial NOo NCJ.-85 



8) VJhat is the effect of the different chemotherapeutic e.geni. 
on maJ-ignant cells in the periplieral blood? 

9) What is tte effect of infection in patients witi. cancer 
on the nmnber and character of uialignant cells in the peripheral 
blood? 

10) Does host resistance developed by a person to his own 
cancer frevjuently prevent living cancer cells in the peripJieral 
blood from growing at another site? How is this resistance 
mediated? 

11) Can one's natural resistance to the growth of his tumor 
be potentiated by immunologic methods? 



Part Be included Yes /x/ No /j^ 

. 36 



;.-,,, NC3:-85 

Pa rt B o Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Pruitt, Jo Co, Hilberg, A. Wc, and Kaiser, Ro Fo: Isolation and. 
Identification of Cancer Cells in Peripheral Blood » Journal of 
Abdominal Surgery, ppo 37-40, May 1959= 

Pruitt, Jo Co, Hilberg, Ao Wo, Kaiser, Ro Fo, Ingraiiam, So Co II, 
Smith, So Jo, and Willoughby, M. B«: Spray Technique for Prepare-tj. 
of Cytologic Specimens for Automatic Scanning Machines, Jo-urnal u-i 
the National Cancer Institute, Volo 22, pp. 1105-1117, June 1959. 

Ejdiibit - "Studies of Cancer Cells in Circulating Blood" was 
displayed at Public Health Service Meeting in New Orleajas, Lao 
in March 1959; at the North Carolina State Medical Association 
Meeting^ Asheville, No C. in April 1959; and at the Clinical 
Meeting of the AoMoAo in Atlantic City, New Jersey, in June 1959o 



Honors and Awards relating to this project: 

T^.e Exhibit listed above won the Certificate of Merit in the 
Preventive Disease Section at the AoMoAo Meeting in Atlantic City 
in June 1959= 



97 



r,.iu-i.i,L.L-'. ;_r, xnvcscxga'oionQ ana 

Individual Project. Report l emoustrations Branch 

Calena.ar Year 1959 2. diagnostic Development Pro/i. 

3. Washington, D. C 



Part A. 



Project Title: Cortisone Study - Georgetown University Medical School 
and Georgetown University Hospital 

Principal Investigators: Dto Dorothea Bo Qiapman, Dto Itonald P. Peysei 

(Dr. Peyser as of NoVo I6, 1959) 

Other Investigators: Mrs. Frances W<, Ondrick, Br. Lo Ho Kyle 

Cooperating Units: Georgetown University Medical School 

Department of Medicine (Dro Lo Ho Kyle) 

Man Years (Calendar Year 1959): 



Total : 


2k 


Professional: 


Ik 


Other : 


1 



Project Description: 

Objectives ; This is a research project with the aim of study- 
ing the effect of cortisone and similar steroids on cancer develop- 
ment and cancer growth. 

It is known that cortisone has an effect on inflammatory 
reactions, infection, and antigen-antihody reactions in animal 
and msuio This effect varies a great deal, depending on various 
factors, for instance dosage and, especially, on the particular 
time of administrations during the course of the disease. The same 
holds true for the effect on cancer and metastasis formation as 
shown by clinical observations in man and experi.iients in anima'' s : 

If more information caii be gained regarding the ...echaniS:.: 
by ■vdiich cortisone mediates its various effects, this should be 
helpful in: 

1) Iiapi'ovement of criteria for institution of steroid- 
therapy in neoplastic and other diseases, 

2) Clarification of relationship and possible similarity 
between cancer and various diseases, 

3) Gain of more infor.ration regarding the basic disease- 
mechanism of camcer. 

Methods Employed : Through cooperative arrangement with the 
Depsirtment of Medicine at Georgetown University selected patients 
axe carefully followed on an inpatient and/or outpatient basis by 
clinical and laboratory examinations (serum proteins, lipoproteins, 

. 38 



<i j-riUiiixv-Tj Oiit:j.Cjj. 



Methods Employed : ( cont » ) 

cholesterol, uric acid). The laboratory studies are being done 
before, dixring, and after administration of cortisone and siudlar 
steroids o An equal number of patients are being selected froiu 
five disease groups (allergy, cancer, collagen diseases, diabetes, 
hypertension) . A control will be established by doing the exaii;- 
inations on normal volunteers also. Thus, data will be collected 
on the differences of various body functions among these patients; 
and on the effect of cortisone on them. In the cancer patients 
especially, the progression or regression of the disease will be 
carefully followed » 

Patient Materieil : The patients are being drawn from the 
various Outpatient Department Clinics as well as from those within 
the Georgetown University Hospital <, 

Major Findings ; During the calendar year of 1959^ 229 patients 
were studied.. On these patients 1,003 serum protein electrophoretic 
strips and kZL lipoprotein strips were processedo No conclusive 
statements can be drawn from the data at this time. 

Significance to the Program of the Institute ; More information 
regarding criteria for steroid therapy in cancer patients is 
highly desirable and sorely needed. There is at this time no 
reliable method of predetermining ■s^ich patients will be improved 
and xdaich patients' conditions may even be worsened by steroid 
therapy., Since there is evidence in animal experiments and 
suspicion in a few patients, who are on cortisone for extended 
periods, that cortisone either causes or accelerates carcinogenesis, 
it is importsmt to establish the true facts of the relationship of 
cortisone to carcinogenesis. It is hoped that data obtained by this 
project may contribute to the solution of this problem as well as 
the problem of the decision i&ether or not an adrenalectomy will 
be beneficial to a given patient. 

Proposed Course of the Project ; At least a three year study is 
planned to select and examine the several hxmdred patients needed to 
gain information ■vdiich would be scientifically and statistically 
valid. 



Part B. included Yes £J No Jx/ 

. 39 



indlviduaf ProjLt Report i*^ i^estl^io^ and 

Calendar Year 1959 , , Demonstrations Branch 

2. Cytology Section 

3. Houston, Texas 



Part A„ 



Project Title; Investigation and Development of Methods of 

Collection of Sputum Specimens ■=•= Houston, Texas 

Ps'lncipal Investigators? Dr„ Heins-ich W, Neidhardt 

Dr„ Bernard L. Yollick 
Dff, Clifton Mountain 
Dffo Fritz R= Dixon 
Dr, William A, Walfces 
Mrs, Earlene To Finnerty 
Mrs, Vera M, Bee 

Other Investigators; Dr, William 0, Russell 

Dr„ R» F, Kaiser 
Miss Anne Baranovsky 
Miss Eleanor Macdonald 
Miss Renilda Hilkemeyer 

Cooperating Units; The University of Texas 

Mo D„ Anderson Hospital and Tumor Institute 

Man Years (Calendar Year 1959) s 
Total s 5 
Professionalf 1 
Other g A 

Project Descriptions 

Objectives; The purpose of this study is to investigate a?«d 
evaluate various methods of obtaining suitable specimens of b'foncbl: 
seCifetions for cytologic examinationo The cytologic technique is 
valuable as a diagnostic tool in primary bronchogenic carcinon!a„ 
However, the successful application of exfoliative pulmonary cytoLo^ 
has been somewhat limited in the past because of the difficulty of 
obtaining suitable specimens of bronchial secretions from patients 
who do not have a productive coughs 

Methods Bapl oyedg Tbs methods which have been considered as 
having possible value Include positive and isegative pressure gss 
machineSs, the Cof=Fiator, chemical agents, inhalation of aerosols „ 
inhalation of various mildly irritating gases, and endotracheal 
suction at time of general anesthesia. 

These patients have not ?5ecessarily had pulmonary disease, and the 
decision as to whether or not a satisfactory sputum is produced is 
based on the presence or absence of columnar epithelial cells and/ 



IOC 



)KIINUAIION SHEET - 1 

or histiocytes in the specimen.. 

Evaluation of each of the techniques considers (1) quality and 
suitability of the bronchial secretions obtained for cytologic 
examination, (2) Adaptability to widespread use in terms of 
acceptance by the medical ps'ofession and patients for use at the 
conanunity level. 

Patient Material; Volunteers, clinic patients, and patiexjcs 
undergoing geneiral anesthesia at the M„ Do Anderson Hospital and 
Tumor Institute. 

Major Findiags g A total of 6,200 sputum specimens have been 
collected^ Over 200 selected patients have received intravenous 
sodium iodide and «ixiother group has received intravenous sodium 
chloride as a controls Statistical evaluation of this group is now 
proceeding. Approximately ilOO sputum specimens have been collected 
using heated hypertonic saline aerosol inhalation, and statistical 
evaluation of this group is now proceeding. 

No recognized untoward physiological reactions to either procedure 
has been founds 

Approximately 670 specimens have been collected by endotracheal 
suction during general anesthesia before and after surgery,. These 
will be evaluated in the near future, 

Ira the overall project, 183 suspicious or positive specimens 
have been obtained from 116 pat lent s^ 

Sigaiflcanee to the Proggam of the Institutes The stvsdy of 
enfoliated cells has been demonstrated to be of great valua in the 
early diagnosis of cervical cancer = The application of the cytologic 
technique to cancer of the luag has been less successful because of 
the difficulty of obtaining suitable bronchial secretions for e^rcaralv 

lesults of these iBvestigations could make exfoliate pulmonsty 
cytology a more useful tool in the diagnosis of lung canceffo It is 
hoped that ultimately the technique will have sufficient precision tc 
be used as a routine screening technique, as is presently being done 
in cervical cytology. 



Proposed Course _of ^ the_Pgo ject s Accumulation of pertinent elata as 
indicated above will continue over a number of months before statistic.; 
evaluation is coiapletedc 

Part B<, Included Ye8/°~7 No.^T 



101 



PHS"NIH i.„ Field iisvescigatioias aud 

Individual Project Report Demonstrations Branch 

Calendar Year 1959 2. Cytology Section 

3o Houston, Texas 



Part Ao 



Project Titles Investigations and Development of Methods of 
Preservation, Processingj and Staining of 
Bronchial Secretions°=Hou8ton, Texas 

Principal Investigators:; Dr, Eeinrich W. Neidhardt 

Dr. Jeffrey P, Chang 
Dr„ Clifton Mountain 
Dr„ Fritz R, Dixon 
DTo William A, Walter 
Mrso Earlene To Finneyty. 
Mr„ Milton Anken 
MeSo Vera Mo Bee 

Other Investigators: Dr, William 0. Russell 

Dt„ R, F» Kaiser 
Miss Anne Baranovsky 
Miss Eleanor Macdonald 
Miss Renilda Hilkemeyer 

Cooperating Unitss The University o£ Texas 

Mo Dc Anderson Hospital and Tuntor Institute 

Man Years (Calendar year 1959) s 
Total s 4 
Professionals 2 
Other s 2 

Project Descriptions 

Objectives; This project is designed to in^rove methods 
of handling and preparing specimens of sputum and/or bronchial 
washings which have been collected for cytologic study utilizing 
a basic science approacho 

Methods Sn^ loye ds Sputum and other specimens are collected 
and saiaples of~these specimens are used for the studies of the 
effects of differes^t methods of preservation, processings and 
stainingo Histochemical techniques are used to study the behavior 
of enzymes, carbohydrates s, nucleic acid, and lipid in exfoliative 
cells as an aid to developing new methods of p5re8ervation, processing,. 
and staining. Several mucolytic agents have been investigated fo? 
liquification of mucous and then cells are concentrated hy me^rane 
filtration and cesstrifugation= Both fresh and fixed sputum specimens 
have been utilizedo Specialized laboratory techniques, such as phase 
contrast microscopy, interference microscopyj end fluorescence 
microscopy may also be evaluated for their usefulness in the laboratcsy 
examination of exfoliated cells, 

102 



ONTINUATION SHKKX ^ ;. 

PatjL ent M at eg i al s Adult hospital patients of tUe M.. Do 
Aoderson Kospital and Tuasor Instituteo 

Major FindingBg (a) By enzymatic digestion, mucous can be 
liquified so that free cells in sputum can be washed in normal 
saline and then subsequently concentrated , This process has 
overcoTiH one of the greatest obstacles in sputum specimen 
p?ocessingc It can be applied to other mucous problems In 
exfoliative cytology as well. 

(b) A cell concentration method has been successfully developed. 
It makes possible examination of a given amount of sputum in 'its 
entirety ; not a random sample as is presently done, over-^night 

or 24 hour specimens may be utilized with all material concentrated 
on a few slides „ Thus the reliability of diagnosis by sputum 
cytology should be tremendously increased. 

(c) The new cell concentration method can be applied to fresh as 
well as formalin fixed sputum according to specific aim of the 
studieSo Obviously, the success in using fixed material for cell 
concentration represents a major advance in exfoliative cytology 
technology, because sputum specimens can be collected at any time 
and at any location. Thus, this method may ultimately be applied 
at a coamunity level for large scale screening of lung cancer. 
Further refinements of the technique are under development and 
clinical evaluation of the technique will begin in the insnediate 
future, 

Signlficaace to the Program gf the Institute? The study of 
exfoliated cells has been demonstrated to be of great value in 
the early diagnosis of uterine cancer o 

It is not possible to make direct smears of all the areas Involve^ 
in the puloMsnary tissue as it is in uterine, specifically cervical 
examinationo Any exfoliated malignant cells from the lungs will 
be diluted by the bronchial secretions when they are brought up as 
sputum specimens and are then diluted by the liquid fixative into 
which the patient espectorateSo The present techniques then make 
only random saaiples of minute fractions of the total spectmens„ 
With the newly developed technique, all cells in 5 mlo of undiluted 
sputum can be concentrated onto 3 membrane filters „ However ;, with 
the conventional method , a large number of slides would be required 
to smear the same amount of sputum. Thus, the new techj>ique aay 
Increase the reliability a hundred fold. This new technique should 
ms^e a significant addition to the diagnostic aids utilized in the 
detection of lung cancer. 

Prop osed Course of the^groiect g Accusiulation of pertinent 
data as indicated above will continue over a number of months before 
statistical evaluation is coapletedo 

LMT B o included Yes/^ No/^ 

103 



PHS-HIH 

Individual Fyoject Repos-fc 

Caleadar Year 1959 



DenvoastrafcioGS iSr; 
2- Cytology Section 
3= Houston s Texas 



Fast A„ 



Project Titles Histologic and Histopathologic Study of 

Exfoliated Bronchial Cells == Houstonj Texaa 

Principal Investigatorss DTc Heiaricb Wo Neidhasdt 

DTo Clifton itouataira 
Bs-c Fritz Ro Dixoti 
Ite, Williasa Ao Walter 
Mss, Earlene To Finnerty 
Mssc Vera Mo Bee 

DTo Willis© Oo Ruseell 
Dro Ro F. KaiaeK 
Miss Anne Basanovsky 
Miss Eleanor Hacdonald 
Miss Reailda Hilkemeyer 

Cooperating Uaitag The University of Texas Mo Do Anderson 
Hospital and Tumor Institute 



Other Investigators s 



(Calendar year 1959); 
Totals 2% 
Fsrofesaioaals ^ 

Others 2 



Project Descriptions 

Objectivess The purpose of this investigation is to deterraina 
the effect of endogenous sxid exogenous factors on the epithelium of 
tracheo-*bso?xhial tree« Histological sections of surgical specimens 
and autopsy material will be examined and correlated %;ith cells 
obtained from the bronchial secretionso This material is being 
obtained from persons without pulmonary diseases parsons raceiving, 
various types of honaone therapy, persons suffering from ni&ltg,iie,r..t 
pulmonary disease, persons undergoing radiation therapy, saokars 
and non-smoker So 

The information obtained from this work will provide a basis fox 
attempting to daviee an accurate and practical classification of 
the normal and abnorsaal characteristics of exfoliated cells from 
tracheo^bronchial tree. 



104 



■>OKTIMJATIO-J? SEEEX - 1 

Methods Ea^loyeds Microscopic examinations are being 
pasfoKmed on whole owgasi. sections of tracheo bronchial tree 
froQ autopsy speclmeas is^rint smeai^s of lung cancers from 
autopsy asid sus'gical speciicens., and eKfoliated cells f^om 
bronchial secretions » 

Epidemiological histories with particular reference to 
inhaled particles (smoke) are being obtained to coic^elate 
with the othe? material o 

Patient Mategjalg Adult clinic aad hospital patients 
of the Mo Do Anderson Hospital and Tuioo? Xnstituteo 

Major Findings; A total of 6200 sputum Bpecimens have 
been collected r, on approximately 2900 patients ^ Detailed 
emoking 'histories have been obtained on 2529 patients, Hhole 
organ sections have been prepared from the tracheo^bronchial 
tsee of 40 iselected cases on whom detailed epideioto logical 
histories (smoking) are evailable. 

Final analysis of ttsis de a has not been coicpletedc 

S ignif 1 c aac e_ to tfeie groggaia.of the Institutes At the present 
time 3 the early diagnosis of bsonchogeaic carcinoma is very difficult. 
because of the inadequacy of the diagnostic methods » A better 
knowledge of the cellular chaages which occur in this disease can 
provide a basis for ia^roving the diagnosis, and Information will 
also be obtained with respect to possible etiological factor s„ 

grogpBg d Course of the_ Projects Accumulation of pertinent dats 
as Indicated above will continue over a nu-aber of foonths before 
statistical evaluation is corapleted. 



PMT 3o included Yes£_./ Mo/_X/ 



105 



PHS-=Nie 

Individual Project RepOiirt 

Calendar Year 195$ 



1 K,!!..L'j iPveKtlgations Rv.a 
Demonstrations Branch 

2, Cytology Section 

3. Louisville, Kentucky 



Part A„ 



P5foJect Title? A study in a general population of cervical cancer 
using vaginal and cervical cytology for pKeliminaTy 
case selection='Loui8ville, Kentucky 

Principal Investigators? Dr=. James C, Djrye, Dr,, William M, Christcp 

and Dr, J^nes E, Parker 



Other Invest igatorss 



Miss Anne Baranovsky 



Cooperating Units; University of Louisville School of Medicine 

American Cancer Society, Kentucky Division, Tn: 

Man Years (Calendar Year 1959) s 
Total % 11 
Professionals 1 
Other s 10 

Project Descriptions 

Objeetives g (1) To provide cervical and uterine camcer i- 
and prevalence data on a jsajor population group; 

(2) To provide data on the 15^20 year age group; 

(3) To correlate tissue studies with results of the exfoliative 
cytology findings on the study population; 

(4) To discover how an exfolistivs cytology laboratory may 
become a self=supporting community medical diagnostic service; a'>d 

(5) To give collect cost study data relative to cervical 
cancer case -identification. 

Methods Emplqyedg (1) Under the direction of the Department of 
Pathology, University of Louisville Medical School and the Tumor Boarc 
Medical Center, University of Louisville, cervical siuears are being 
obtained by the swab method j prepared and examined according to the 
usual Papanicolaou technique on the adult female population. Vaginal 
aspiration smears will be obtained for correlation and comparison vif- 
the cervical swab smears on the Louisville General Hospital patieifits 
on women screened in industry. All smears are screened and checked b 
teclmlcians with a pathologist reviewing all suspicious and positive 
slides and deciding the cytologic evaiuationo All suspicious or 
positive cases have tissue studies. 

(2) The cases found by biopsy to be positive will be treated oi; 



106 



OHTIMJATXON SHEET - I Serial. No. HCI->U) 

Method s En^loyed; ( cont » ) 

referred for treatment, 

(3) A histopatboiogy laboratory is niaintaiaed for studying 
serial sections on those cases in which the original routine 
histology did not correlate with the cytologic findings.. 

Listed belov; are steps in the established procedures 

(a) History card designed by the Field Investige-tione and 
Demonstrations Branch of the National Cancer Institute will be 
cooqpleted on each patient tasted, 

(b) Recall of all patients who have questionable smears »- 
repeat or biopsy, 

(c) Routine recall at one year of all those tested. 

(d) Transfer of all data to IBM punch cards for tabulatio? 
and analysis.. 

Patient Materialg There ate more than 200,000 women in the 

study areao About 13% are non°\;'hite. 

Major Findings; 

GERVIGM. SWAB TECHNIQUE 
PRELIMINARY F INDINGS TO JULY 1. 1959 

1st Screening, „ <, .. , » n , „ o59 ,687 
2nd Screeningo , , , , .. = . , » 16 s929 
3rd Screening. o , , . . o , , , 2 , 752 
Other Repeat Tests, . , i ., 7,384 
Total Tests 86,752 

First Screening of 59j63I womes^ as to Sources 

Louisville General Hospital & ClinicSo «.. 14,637 

Private Patients „ . „ .. , « .. » » . , . o » = . » o . = . o , » » 33 j 315 
Industry .,,,,, c » o o,, o ,. o».>« c = p o c » ^^ c « oo = » 11 , ?3:) 

Results of First Screening (59,687 women)? 

Total biopsies Requested, „ . „ <. <. , » = o „ » =680 

Biopsies Negative, „ -, o » » o o = » . c o , , ,16? 
Biopsies Not Doneo „ „ , , » = « , » » o , » » o ?1 
Biopsies Abnormal c », oo, ^ .» ..,-,.>« = AA2 



107 



li'j^iric;;! 5HEET - 2 -^^.^^a-i ^/^ i-ICi'-i-^ 

M a.1og_Fi nd£agsg Ccoat„) 

Dysplasia, „. -,,<,, oc„,, o,„», o ,,-.<.,,. „ 128 
Intraepithelial Carcinoma^ „.,,,=, „ 163 
Squamous Carcitiomao , . o . » - » , . <. , o , » . 129 
Adenocarcinoma of Cervix, ^ ^ ^ , , , » , .. 3 
Adenocarcinoma of Endometrium, , , ^ o II 
Unclassified, recurrent and 

other cancers o „ o , •, = .,,,..,,«., o- „ 8 

Significance to th e Frogram of the Institute; The results of tfei^ 
study will be added to the information being obtained in otlias cyfcolog: 
studies. Consolidated data are expected to furnish insight into patho- 
genesis of uterine cervical cancer, course of development of the lesion 
and its incidence and prevalence in the adult female populationc 

Propose d course of the P rojec ts Tfea direct operation of this 
project y>s.n terminated on June 30, 1959, A contract is supporting 
collection of final data on the study group through fiscal year l^^''^' 



Part B included Yes /X / Mo/ / 



108 



Individual Project EepoK't Serial Hu,. „__^^i^.L 

Calendar Year 1959 



£=^X^° Honors s Awards and Publications 

Publications other than abstracts fsom this projects 

Christopherson, W„ M. and Parker., Jo S.s 

A Study of the Relative Frequency of Cascisioma of the 

Cervix in the Kegs'Oo Presented Inter-Sociefcy Cytology Council 

Hov, 19, 1959 



Honors and Awards relating to this project: 



109 



PaS-NIH 

'alentor Year 1959 2 Cytology Section 

3o Loy..i=-"' " " V . Kentucky 



Part Ac 



Project Title: A uterine cervical cancer morbidity study in 
Loiiisville, Kentucky 

Principal. Investigators: Dr. William Mo Cliristopherson, 

MrSo Winifred M= Mendez 

Other Investigators: Miss Mo M<, Bouser, Miss Anne Baranovsky^ 

Dr. James Eo Parker 

Cooperating Units: University of Louisville School of Medicine 



Man Years (Calendar Year 1959): 
Total : 3? 
Professional; 
Other 



1. 

2 

3 



Project Description: 



Objecti¥es : l) To determine the impact of cervical cytolog;y 
on the incidence of cervical carcinoma in a large urban area >dien 
the cyt.ologic examination is offered to women througb. their femiily 
physician, the heeilth facilities in industry and the clinics in 
the city hospitsilo 

2] To determine the incidence of symptomatic cancer of the 
cerTTix among the non-cy-fcologically tested population of women ir 
the Louisville areao 

3) To provide a baseline for comparison and evalxiation of 
cytology results obtained from the Louisville Cancer Survey Px-oject. 

h) To provide adequate assessment of the meaning of case- 
finding or incidence rates for cancer of the cervix on successive 
screenings ^ 

5) To help discover symptomatic cases between screenings 
among -women in a cervical cytology study, some of -vmich may have 
been missed during the previous screeningo 

6) To help clarify the natural history of the disease o 

Methods Employed ; 1} Identification aaad clinical description 
of aJLl cases of uterine cancer diagnosed, treated, or deceased in 
Jefferson County for the years 1953-5^5-55^ t© establish a baseline 
of uterine cervical cancer incidence in the population prior to 
the Cancer Survey Project » 

110 



2) Identification auid clinical description of all cases of 
uterine cerxrical cancer diagnosed, treated, or deceased in 
Jefferson County during 1956-57-58, the years during which the 
Cancer Sxarvey Project was functioning in order to study the 
effect of a cytology project on tiie incidence of cervical cancer o 

3} Identification and clinical description of all cases of 
cervical cancer diagnosed, treated, or deceased for the years 
1959-60-61, the years following tke termination of the cytology 
project in order to study the result of the cytology project on 
the incidence of cervical cancer and the effect of cervical, 
cytology on the course of the disease and of survival rates in 
the cytology population » 

k) Identification of the uterine cancer cases will he 
accomplished hy viriting all the local hospitals where uterine 
cancer is diagnosed or treated, all private pathologists and 
radiologists o Case records will be summarised and matched 
with records of patients in the cytology project population.. 

All death certificates registered in Jefferson County listing 
utei'ine cancer as the primary or contributory cause of dea-th will 
be listed and will fee matched with cases identified from all 
other sources and with patients in the cytology population » 

5) Overall incidence rates for cervical cancer and rates 
for the cytology and non- cytology populations wi3JL be studied 

for the periods prior to the cy'fcology project, during the cytology 
project and post-project period. 

6) Survival rates for uterine cancer cases in the cytology 
and non-cytology populations will be compared. 

7) A card was designed for abstracting case records, ajid a 
marginal punch key sort card is being prepared for collating the 
information from all soiorces on each case. 

Patient Ifeterial. ; All uterine cancer cases diagnosed, treated, 
or deceased in Jefferson County, Kentucky. 

Major Findings; The study was initiated in June 1959 and, suf- 
ficient data is not yet available for analysis. Pathology reports 
have been listed from aJil the hospitals and private pathologists 
for the period 1953-58o Abstracts of case records have been 
completed in three large hospitals and one small hospital for the 
six-year period o 

Significance to the Program of the Institute ; The morbidity 
study should assist in evaluating cervical cytology as a tool for 
eas'ly diagnosis of cervica3. cancer. On the basis of patient records 
the value of cervical cytology in the initiation of early treatment 
and consequent anticipated improved survival may be appraised o 

111 



Sigrtlficance to the Pyogram of the Institut e; (cont . ; 

In conjimction with ma,terial from other simileir studies 
the data from this project vill help to clarify the natural 
history of the disease. 

Proposed Coturse of the Project ; Continued collection of 
ilata on uterine cancer cases in the study area for the projected 
years o Recording the collated information on marginal ptmch key 
sort cas=ds and analyzing the data as sufficient material is 
organissedo 

Adding case history data for the years 1959'60-6l as the 
patient records become available » 



Part Bo included 'ies /__/' No JX/ 

112 



PHS-NIH 

Individual Project Report 

Calendar Year 1959 



Serial No. NCI-90_^ 

1= Field Investigations and 
Demonstrations Branch 
2, Cytology Section 
3,. Madison, Wisconsin 



Part A, 



Project Title; 



A study in a general population of cervical cancer 
using vaginal cytology for preliminary case selcctior 
Madison, Wisconsin 



Principal Investigators? Dr, Willianj D„ Stovall, Dr„ Robert Steinkraus, 

Mrs,, Korma V„ Arvoldo 



Other Investigators: 



Miss Anne Baranovsky 



Cooperating Unitsj University of Wisconsin Medical Center Laboratoify 
of Hygiene 

Man Years (Calendar Year 1959); 
Total ? 14 
Professional? 2 
Other ; 12 

Project Descriptions 

Object ivesg This study is designed to explore the feasibility 
and practicability of bringing the cytologic procedure to rural women 
in a four teen-county area utilizing practicing physicians to gather 
the necessary specimens and to submit them to a central laboratory 
for interpretation. The study will provide data on the prevalence and 
incidence of carcinoma-in-situ and invasive cervical cancer in a cotal 
rural population of approximately 350 ,000 , with 109,000 women ovei- 21 
Data are also to be collected concerning the type biopsy and the treat = 
ment performed on the malignaacies encountered„ 

Meth ods Enjoye d s Through a cooperative arrangement between the 
University of Wisconsin Medical Center and the FIDB a project has been 
established which will provide study material and laboratory inter- 
pretation of vaginal and cervical smears. The study is under the 
direction of Dr. William D„ Stovall of the University of Wisconsin, 
Vaginal and cervical smears are collected by over 300 participating 
physicians in 14 counties and mailed to the State Laboratory in kits 
which have been provided. The results of the smears are reported 
directly to the local physician. Mild atypicelities are followed by 
subsequent smears. Cytologically positive and suspicious cases are 
advised to have biopsies performed^ Tissue specimens may be sent to 
this laboratory or to any pathologist the local physician chooses. 
In all cases we request a complete report from the physician concern- 
ing type biopsy and subsequent treatment for our recoicds. The data 
obtained are recorded in the appropriate medical scatiotical form 18l9-Ci 



113 



CONTINUATION SHEET = 1 Serial No., NCI^90 

Methods Eg^loyedg ( cont o ) 

en5)Ioyed by the other cytology units and microfilmed for coding in 
Bethesda. Postcards reminding patients to come in for their annual 
screen3.ng are sent to physicians' offices at the appropriate intervals. 

Patient Material; Patietsts of private, cooperating physician? 
in lA counties of Wisconsin, 

Major Findings; The unit began its activities on July 26, 1956 „ 
As of November I, 19598 lOAjSGl screenings have been done representing 
68s,406 cases. Of ls041 cases having abnormal cytology 406 cases have 
proven positive on biopsy;, on subsequent smears or biopsy 391 have been. 
proven negative and 244 need further smears or biopsy. There are 33 
cases kno\m having false negative cytology. Approximately 20% of cases 
with atypical smears j, 507e of cases with suspicious smears and 80-907.. 
of cases with positive smears eventually have positive biopsies.. 
Slightly more than half of the cases of carcinoma- in situ have no 
evidence of malignancy at the time of hysterectomy if a cone biopsy 
had been performed prior to treatment o Several articles are under 
preparation discussing the accumulated cytologic, biopsy and treatment 
data. 

Significance to the Program o f the In stitutes The results of this 
study will be added to the Information being obtained in other cyto- 
logical studies. Consolidated data are e3q>ected to furnish insight 
into pathogenesis of uterine cervical cancer, course of development 
of the lesion^ and its incidence and prevalence in the adult female 
population. 

Propose d Course o f the Project ; Data collection on patients 
will end December 31, 1959, 



Part B, included Yes/ / No/X / 



IIH 



PHS-NIH 

Individual Project Report 

Calendar Year 1959 



Serial No. _._„„NCl29l_ 
1, Field InvestigatioDs and 
Demonstrations Branch 
2« Cytology Section 
3o Madison, Wisconsin 



Part A„ 



Project Title; A retrospective study in a selective patient group 

of cervical cancer using vaginal cytology for prelimisar 
case selection =■ Madison j Wisconsin 

Principal Invest iga tor ss Dr^ William D,-, Stovallg Dr, Robert Steinkraus, 

Mrs, NoriHa Arvoldn 



Other Investigators? 



Mi S3 Anne Baranovsky 



Cooperating Unitss University of Wisconsin Medical Centers, Laboratory 
of Hygiene 

Man Years (Calendar Year 1959)? 
Total ; 2% 
Professionals \ 
Other s 2 



Project Descriptions 



jectivess This study is designed to add data to the preseRt 



cytology study of a rural Wisconsin population by analyzing the 
records of previous smears submitted to this laboratory during the 
past thirteea years. 

Methods Employed ; The records of all vaginal cytology screenings 
for the last thirteen years at the State Laboratory of Hygiene, Madison, 
Wisconsin^ have been sorted and analyzed with particular attention to 
those women who reside in the fourteen counties presently under study. 
The previous records of these women have been incorporated with the 
present survey and recall cards have been sent out at the appropriate 
times o The laboratory space, supplies, equipment, and staff are 
Identical with those presently in operation at the Madison Cytology 
Project o 

Patient Materials Material used in this study consists of 
specimens sent in by private physicians in 14 counties in various 
locations in the state of Wisconsin, 



115 



CONTINUAIION SHE' :;eslal No- , j jgj^lgj-. 

Major Findings; ApproxliiBately ?OjOOO cytology reports have been 
alphabetizedg sorted and filed„ Of these cases, II ,,575 lived in the 
14 survey counties and 3630 cases were found to have had smears in our 
survey. Recall cards were sent out on 13,945 cases in an effort to 
incorporate them in our present surveyo We have received smears on 
3059 of these cases.. 

Signifi cance to the Program of t he Institute; This study, by 
merely incorporating past laboratory records j will have the effect 
of adding a considerable number of patient years to the cancer 
experiences of the study population., Ifliat it may insert in the way 
of sampling bias atill needs evaluation, 

Proppsad Course of the Project ; This project will end early in 
1950, 



Part B. included Yes / J Wo/X / 



116 



PHS KIH 

Individual Project S.eport 

Calendar Year 1959 



- "°' ^..^y^ryj*^ 

: laid Investigations and 
Demonstrations Branch 
Cytology Section 
Hagerstown, Maryland 



Part A, 



Project Titles 



Field evaluatioB of the perfomnance of an electronic 
cytoanalyzer. Investigation of new methods of 
preparing vaginal cervical matetrial for automatic 
scanning, Hagerstown, Washington Countyj, Maryland 



Principal Invest igatorss 



Other Investigators; 



Cooperating Units; 



Dr, William B„ Courtney, Dr„ A, W„ Hilberg,, 
Dr, S= C, Ingraham, II, 

Ifea, Gloria S, Davis, Mr, Paul Montague, 
Miss Sarah Jo Smith, Miss Anne Baranovsky, 
Miss' M.. M. Bouser, Dr, to F, Kaiser 

Golumbusj Ohio, Cancer Survey Project 
Walter Eeed Army Hospital 
Johns Hopkins Hospital 
Airborne Instruments Laboratory 
Hagerstown J Washington County Health Dept. 

Man Years (Calendar Year 1959); 
Total ; 4 3/4 
Professionals I 3/4 

Other s 3 

Project Description; 

Objectives g To evaluate the performance of the cytoanalyzer and 
to develop new techniques for preparation of smears for in5)roving the 
performance of the cytoanalyzer. 

Methods Em ployed ; Cytoanalyzer performance is coopared to routine 
screening using results of the Cytology Laboratory, Vaginal specimens 
are subjected to a variety of procedures and the resulting preparations 
are evaluated for characteristics which will improve cytoanalyzer 



?atlg!t vt_M.5t errialf, F.'Stie.txt^ are c'ravn ft^otja Walter ?.e.ed Army 

Fto?Ti:lt.nl, Jo"hri''. Tio?''^:It^^ aoiT'i£a\, &v<.d Colura>«us Cancer "urvey Project- 



117 



CONTINUATION SHEET - 1 Serial No, NCi-94 



Major, Fiadings : Fitrst clinical trial of the cytoastalyzet 
constieuted aa attends ito evaluate possibility of automatic, electronic 
scanning of smears. Also, it accentuated deficiencies in prog5fam ar.a 
accentuated needed improvements ^ and the running of a mass clinical 
trial-, Major improvements were addition of automatiCj self^focusing 
optical system and inproved method of smear preparation by modified 
drop technique. Sieve and spray technique of smear preparation cause? 
significant cellular distortiono 

At present a clinical trial of 10,000 cases is in progress tc 
evaluate present screening abilityo 

Signific ance to the Prcgram of the Instifcuteg Application of a?. 
electronic scanner in BO??trng eytologlcal preparations, if successful ,, 
can facilitate rapid mass cancer screening with earlier case detection 
and treatment J probably resulting in reduction in cancer morbidity 
and mortality^ 

Proposed Course of the Project ; Engineering designs are planned 
to stabilize and standardize circuiting and to provide automatic slide 
feed as well as medical developmeKit to standardize specimen collect 
tionj preparation and staining methods, "These developments are all 
needed to have a usable working model of the cytoanalyzer in operational 
state a major portion of the time. 



Part B included Yes /X / No / / 



118 



PHS'NIH Seyial No, NCI=94 

Individual Project Report 
Caleadar Yea^ 1959 



Pagt_B. Honors, Awards, and Publications 

Publications other than abstracts from this project; 

Pruitt, J, Co, Hilberg, A. Wo ^ Kaiser, R, Fo , Ingraham, So C. , 11, 
Smith, So J. and Vfilioughby, Mo B,s Spray Technique for Preparation 
of Cytologic Specimens for Automatic Scanning Machines,, Journal of 
the National Cancer Institute, Volo 22, ppo 1105=1117, June 1959o 

A e paper "Cytoanaiyzer Field Trials" by Dr8„ W. Bo Courtney, 
R, Fo Kaiser, A, Wo Hilberg, S, Co Ingraham, II, and Mary M., Bouser 
was read at the November 1959 meeting of the Inter-Society Cytology 
Council in Detroit, Michigan, and will be published in the Trans- 
actions of the Seventh Annual Meeting o o , „ 

In January and February informal talks on Cancer and also 
about the Cytoanaiyzer were given before small groups such as 
the Presbyterian Young Married Couples and the Hebrew Men's Group„ 



Honors and Awards relating to this projects 

None 



119 



PHS-NIH 

Individual Pi-oject Report 

Caaendar Year 1959 



NCI-103 



Serial Hoo 

lo Field Investigations and 
Demonstrations Branch 

2. Cytology Section 

3. Eager 5 town, Maryland 



Part A< 



Project Title: 



Examination of sp-utum specimens from uranii-un 
miners. Investi^tions to evaluate cytologic 
morphology in a controlled group with suspected 
carcinogenic occupational environment - 
Hageretowii, Maryland 



Principal Investigators: Dr. William B= Courtney 

DTo a.- Wo Hilberg 



Other Investigators : 



Miss Sarah Jc Smith, Miss Jean Keller, 
Mrs. Sarah Jones, Dr. Victor Eo Archer 



Cooperating Ifcits: Colorado Plat«au Industrial Physicians 
Man Years (Calendar Year 1959): 



Total 

Pi'ofessional 
Other 



3 3/4 

1/4 
3 1/2 



Project Description: 

Objectives : To evaluate morphclogj'' of cells from the bronchial 
tree fo\ind in sputum specimens of uranium miner©; also, to develop 
a technique for transporting and processing the specimen., 

Methods Employed : Field collection of sputum specimens, field 
processing, and shipment to the laboratory for staining and mor- 
phologic evaluation by technicians, cytologists and pathologist)?. 

Patient Materia l: Uranium miners on the Colorado Plateau ^ 

Major Findings : None to date., 

Significance to the Program of th e Institute: By evaluating 
the findings from the above it should be posBible to relate the effects 
of carcinogenic occupational hazards in uranium miners to the extent 
of morbidity and mortality and to determine the extent of related 
morbidity and thus place proper emphasis on the need, if any, for 
proper protection., At the same time it will provide information con- 
cerning other pathological changes of the lower respiratory tract.- 

Proposed Cotirse of the Project: To screen sputum specimens as 
a malignant case-finding project as well as to evaluate morbid tissue 
reactions over a period of time. 



Part B. included 



Yes /T 



No ,/x7 

120 



PHS -KIH 

Individual Project Report 

Calenda?r Year 1959 



Serial No. NCI-95 

1, Field Investigations and 

Demonstrations Branch 

2, Cytology Section 

3, Philadelphia, Pennsylvania 



Part A, 



Project Titles A study in an industirial population of cervical cancer 
usiEg vaginal cytology for preliminary case selection 
Pliiladelphiaj Pennsylvania 



PE'incipal Investigators; 



Other Investigators; 



Dr, I„ N. Dubin, Dr, Mo R. Dunn, 

Dr„ Bernard Behrend, Dr, M, D<, Pettit, and 

De„ William A. Walter 

Dr, S„ C, Ingraham, II, Dr„ A, W, Hilberg, 

Miss M, M, Bouser, Dr. R, F„ Kaiser^ 

Miss Anne Baranovsky, and Dr„ S, A., Brenner 



Cooperating Unitsj Woman's Medical College of Pennsylvania 

Departments of Pathology^ Preventive Medicine j 
Gynecology and Oncology, 

Man Years (Calendar Year 1959) s 
Total ; 21 ^ 
Professional; 3 k 
Other J 18 

Project Description; 

Objectives; Forty thousand employed women in the Philadelphia area 
will be examined annually by means of vaginal cytology smears for three 
yearSo This study will provide data on (1) the detection of cervical ca5->c 
in the early stage, and (2) various factors in the etiology and patho= 
genesis of the disease, such ass 

(a) prevalence and incidence of cervical cancer 

(b) inter=relationshtp of such lesions as atypical metaplasia, 
intraepithelial carcinoma and invasive carcinoma of the cervix 

(c) whether intraepithelial carcinoma always progresses to invasive 
carcinoma, and over what period of time 

(d) whether intraepithelial lesion seen during pregnancy is the same 
lesion as in the non^pregnant woman, and 

(e) role of age,, ethnic background, socio-economic status, 
gynecologic and obstetrical histpry^ and geographic localea 

f 
The reasons for studying en?>loyed women are? 

(a) this is a stable group s with little turnover of population 

(b) the economic status is more determinable 

(c) this group is better organized, and it is easier to get to 
and work with this group 

(d) one can utilize already existing occupational medical health 
services 



191 



ONTINUATIOK SHEET = 1 Serial No, NCl-95 



Met jipjds, Egployedg This project is the product of a coopet-auiwe 
arrangement between the National Cancer Institute, Field InvestigatioEis 
and DejnonstJrations Branch, Cytology Section, and the Woman's Medical 
College of Pennsylvania, The National Cancer Institute provides the isnmer 
ate professional, techKsicalj, and administrative staff,, as well as the 
necessary supplies and equipments The Woman's Medical College p«:ovides 
the laboratory space, with its permanent fixtures and maintenance^ 

DTo I, No Dubin^ Professor of Pathology of the Woman's Medical 
College, is the Director of the Project, There is an Advisory Committee 
representing the participating departments of the Woman's Medical College 
(viZj the Departments of Pathology, Gynecology, Oncology, and Preventive 
Medicine) 3 The Philadelphia County Medical Societyj the Philadelphia 
Branch of the American Cancer Society 9 the College of Physicians, the 
Philadelphia Pathological Society, the National Cancer Institute,, a 
clinical cytologistj and additional representatives of local teaching 
institutions. 

There is also a Special Committee on Preventive Medicine, con?>rising 
physicians from the Woman's Medical College, Philadelphia physicians 
active in occupational medicine, and a representative of the Industrial 
Nurses AssociatioBo 

Contact with employed women is made by the physicians on the staff 
of the Projects, through the Occupational Physician of a given industry- 
There is no publicity £0 the lay public at large. The enployees are 
given the choice of 

(1) going to their private doctor for a complete physical 
examination and vaginal cytology smear (to be sent to the Cytology 
Laboratory) , or 

(2) having a smear taken by trained personnel of the project, 
at the industrial site. In all cases a card with pertinent data is 
sent to the project with the smears » All reports on the cytologic 
specimens go to the woman's private doctor, who in turn sends a report 
to the patient and acquaints her with its significance. Patients with 
suspicious smears are referred to their private physicians who base 
definitive diagnosis and treatment on accepted clinical and biopsy 
studies, Follow°up data is sought through the cooperation of (I) personne 
of the projects, (2) occupational physicians and nurses, (3) private 
doctors, and (4) pathologists in other institutions who make available 
slides from biopsy specimens for study by the pathologists of the project. 

The data obtained is coded and recorded in a medical statistical 
form (1819=9) as utilized by other cytology units. These cards are 
microfilmed for statistical analysis by the biostatisticians in 
Bethesda, 



122 



ONTINUAflON SHEE Serial No„ NCI -93 



Patient Material; Eiii|>Ioyed women iis selected iadustriss in fc^ne 
city oF¥Sni3e^IOrweire utilized during the first year of operation, 

Duritig 1959 3 activicies were expanded to include the surrounding 
counties of Bucks ^ Delaware and Moatgomery , 

Ma Jog Fi ndings g During the first two years of active screeningj 
249I4O patients were examined by vaginal smears. There was s tora.'i o5 
4,389 wosian screeaed for the second timeo Of these numbers, 195 were 
considered as suspicious or positive for malignancy. Of these 195 cases* 
76 have had biopsies reported to uSo Of these ?6, 14 had invasive car-= 
cinoma of the cervixj, 24 had carcinoma='in=8itu, 14 had atypism of cervical 
epithelium, 19 had inflemmation of the cervix^ 1 had endrometrial car- 
cinoma and 4 had inadequate biopsy specimens. 

Of our total cases 1078, have shown trichomonads on the smears. In s 
special study, the prevalence of invasive carcinoma of the cervix was 
found to be 9,5 per IsOOO in a series of subtotal hysterectomy caseSo 
This was the same prevalence as occurred in our control group when 
corrected for age and racial and ethnic background. 

Significance to the Proggan of the In stitutes The results of this 
study will be added to the information being obtained in other cytologic 
studies Consolidated data are expected to furnish icsight into patho^ 
genesis of uterine cervical cancer, course of development of the lesion 
and its incidence and prevalence is the adult female population. 

Propose d Course of the Project ; A five-year program is antici- 
pated in order to examine 40,000 women annually for three years,. 



gart B, included YesjO No/Tj 

123 



Serial No. MCl>9-» 
Individual Project Report 
Calendar Yea^ 1959 



Part B. Honors 8 Awards and Publications 

publications other than abstracts from thi^ project; 

A paper by Marvin Wo Stafford ^ Supervisory Technician, entitled 
"An loproved Method of Labeling the Frosted-End Microslides Used in 
Exfoliative Cytology" has been accepted for publication by the board 
of editors of "The Technical Bulletin of the American Journal of 
Clinical Pathology," 

Dr. Marvin R. Dunn^ Assistant Director, presented the paper, 
"The Prevalence of Carcinoma in Cervical Stunts" at the 6th Mnual 
Meeting of the Inter=Society Cytology Council in Detroit, Michigan, 
November 19-21, 1959, 



Honors and Awards relating to this project; 



124 



f.h3-k:ih 

Individiia.1 Px-oject Report 
Calendar Year 1959 



Serial No. NCI-96 

1 Field Investigations and 
Demonstrations Branch 
2. Cytology Section 
3„ Ssua Diego, California 



Part Ac 



Project Title: A study in a general population of cervical cancer 

using cervical cytology for preliminary case selection 
San Diego, California 

Principal Investigator: Dro John Eo Dunn, Jr» 

Other Investigators: Dr= Purvis L. Martin and Collaborators 

Cooperating Units: Gynob Clinic, Sajx Diego, California 



Man Years (Calendar Year 1959^ 
Total 

Professional; 
Other 

Project Description: 



1 

5 



Objectives ; One objective is to extend the activities of the 
cytology work of the Gynob Clinic to include indigent women of 
San Diego City and County» This is a trial of facilitating 
coverage of a total, population group for cytological testing by 
supplementing private laboratory re source So 

Another objective is to make available data on this segment 
of the population regarding cervical cancer for study along with, 
the data already available from examinations of seme i*0,000 
private patients. 

Methods Employed : The Gynob Clinic is composed of a gs'oup 
of gynecologists, operating a cyt.ology laboratory jointly, ^o 
ai'e thorou^ly convinced of the value of cervical cytology in 
cervical cancer case-identification. They have operated their 
laboratory at minimum fees to make the service as widely avail- 
able as possible to other physicians as well as their own group « 
They have not been able to extend their services to indigent 
women. Supplementation of Gynob Clinic Laboratory staff by 
the Field Investigations and Demon.^trations Breinch has made 
possible the extension of these services to the indigent portion 
of the population. 

The cytology laboratory has excellent professional and 
technical supervisory personnel and extension of the case -finding 
program to the indigent population is being accomplished throu^ 
the addition of federally employed screening technicians. 



1 oc 



iiWATION SHEE- NC: 



Patient Material : A13. adult fema3,e private patients and 
indigent adult women of San Diego County througih their usual 
source of medical service » 

Major Findings ; The data from the cytological experience of 
the Gynoh Clinic up to 1955 are in the process of analysis » 
These data include the findings from 33,750 women examined 
initially; 9,726 examined a second time; and J*, 213 a third timeo 
The results of the ansuLysis should be very interesting for com- 
parison with other similar studies that have been published or 
soon will be.. For example, the rate of finding carcinoma- in-situ 
seems to be hi^er them Nieburgs' results for Floyd Coxmty, 
Georgia, or unpublished results from Memphis, Tennessee o A manuscript 
analyzing these data will soon be submitted for publication = 

Significance to the Pro-am of the Institute : The atudy will 
provide data on the uterine cytological findings using direct 
cervical specimens » 

From the standpoint of cancer control, it will be of 
interest to learn whether private physicians can effectively 
promote a pro-am of community case-identificationo 

Proposed Course of the IVoject ; The cytology project for 
all of San Diego County is rapidly gaining momentum o A central 
registry has been established ajid the mechanics of processing 
cytology reports and photostating for Field Investigations and 
Demonstrations Branch, Cytology Section, and statistical study 
have been worked out« The program is being gradually expsinded 
to a community-wide program tirging all women to obtain periodic 
cytological examinations throu^ their physici£uis= A program of 
both physician, and lay education will be developed « 



Part B, included Yes [Tf No rj 



1?R 



Calenctar Je&r 1959 

Part B o Honors, Avards and Publications 

Publications other than abstracts from this project: 

Dunn, Jo E«, Jr., Slate, To Ao, Merritt, Jo Wo, and Martin^ Po L.: 
Finding for uterine cancer from one or more cytologic examinationa 
of 33^750 women o Journal of the National Cancer Institute, Volo 23, 
po 3, September 1959° 

Dunn, Jo Eo, Jro: The epidemiology of cervical cancer as revealed 
by cyto^-ogyo (To be published in JoumaJL of the International 
College of Surgeons o) 



Konora and Aisards relating to this project; 



127 



PKS-NIH 

Individual Project Reporfe 

Calendar Year 1959 



Serial No, tjCIjlO'^^ 

1, Field Investigations and 

Demonstrations Branch 

2, Cytology Section 

3, Winston-Salem, Nurth Carolina 



Project Titles A detailed investigation of Gastric Cancer 

utilizing Exfoliative Cytology for case selection - 
Winston=Salem, N. Co 

Principal Investigators; Dr. Robert P„ Morehead, Dr. Carl A, Tattory, 

Dr. Hugo C, Pribor, Dr„ David Cayer, 
Dr„ Marcus Fc Sohmer, Miss Mary Eo McGovern 



Other Investigators^ 



Dr, R, Fo Kaiser, DTo S, C, Ingraham, II, 
DTo a. Wo Hilbergs Miss M„ M, Bouser 



Cooperating Unites Bowman Gray School of Medicine of Wake Forest College 

Man Years (Calendar Year 1959) 
Total s 10 
Professionals 3 
Other g 1 

Project Description 

Obieetivesg The field investigation using the methods of exfoli 
ative cytology for study of human gastric cancer is engaged first in 
evaluating known methods of gastric specimen collectioDj then ia study- 
ing pathogenesis « course of progression and response to therapy of 
gastric cancer > 

Methods Employedg Methods of approach include a study of patients 
preaes&ting symptoms referable to the stomach as well as those 
without synqj tomato logy who are found to be achlorhydric by tubeless 
gastric analysis^ In this manner a study of cellular morphological 
details in a variety of gastric lesions as well as comparison of these 
details to normal gastric epithelium and known gastric cancer cells is 
possible. Biochemical studies on gastric secretion are also being 
investigated in an attes^t to facilitate gastric cancer patient 
identification from the general populatiosio In additions, hematological, 
physiological and anatomical methods of possible preselection of human 
patients are planned. 

Patient Material; Patient material is derived from the inpatient 
and outpatient population of the Medical and Surgical Services of the 
Baptist Hospital in Winston Salem, North Carolina. 



128 



CONTINUATION SHEET - 1 Serial No. NCI- 104 



Major Fin diagsg The study of exfoliative cytology is proving 
to be a valuable tool in the diagnosis of gastric cancer c It appears 
to have merit in confl&uaing the presence of malignancy in those cases 
which are equivocal by other methods of diagnosis. It should provc- 
helpful in detecting gastric cancer before it is apparent by the 
routine methods of diagnosiSo 

Significance to the P?eogram of the In stitute; By developing 
methods for detecting and diagnosing gastric cancer in its early stages 
this study wills if successful ^ offeif a means whereby this type of 
cancer can be more readily controlled in the general population. 

Proposed Course of the P roject ; This project is designed to 
run a 2 to 5 year course depending on the scope of its early findings. 



Part B, included Yes/y^/ No 



129 



NIH-FHS 

Individual Project Report 

Calendar Year 1939 



Part B, Honors, Awards^ and Publicat ons 



Publications other than abstracts from this projects 

Hunts Wo B., Jr„; Cayer, D, ; and Kalnins, Z. As Gaistric 
cytologic findings in 100 patients with gastroiafceetlnal symptoms, 
presented at a meeting of the North Carolina State Medical Society 
in May 1959- 

A paper entitled "Evaluation of Diagnostic Procedures in the 
Clinical Diagnosis of gastric cancer by N. F, Sohmer, D, Cayer and 
Zo A, Kalnins was read before the Section on Gastroenterology, 
Southern Medical Association, Atlanta, Georgia, Nov, 17, 1959, 



Honors and Awards related to this projects 
Nosie 



1 .'^n 



Individual Project. Report 
Calendar Year 1.959 



Part Ao 



Serial No.__jr- 

1 Field ).niArei>;. i.v,.tif. i . 

Demonstrations !&■ 
1~. Cytology Section 
3. Houston, Texas 



Project Titles Studies on distribution of exfoliated cancer 

cells within tracheo- bronchial tree, correlation 
with the cellular niorphology of the same tumor 
from imprint smears ^ and persistence of cells in 
bronchial secretions of patients after surgical 
extirpatiora of the tumor-^HoustoOs Texas 



Principal Investigators? 



Other Investigators; 



Cooperaicirag Uaitss 



Dr, Heinrich W. Neldhardt 
Drc Clifton Mountain 
Dr. Fritz R, Dixon 
DTo William A. Walter 
Mrs, Earlene To Finnerty 
Mrso Vera M,. Bee 

Dr. R„ Fo Kaiser 

li^c William Oc Russell 

Mr I Ben Carroll 

Miss Anne Baraitovsky 

Miss Eleanor Macdonald 

Miss Renilda Hilkemeyer 



The University of Texas 

M„ D„ Anderson Hospital and Tumor Institute 



Man Years (Calendar Year 1959) s 
Total % 3/4 
Professionals 1/4 
Other % 2/4 

Project Descriptions 

Object lyes g The purpose of this esqperlment is to determine 
the distribution of exfoliated cancer cells in major bronchi of 
extlErpated lung tissue., the cellular morphology of the tumor 
itself by imprint smears , and the persistence or recurrence of 
malignaiiit cells in the bronchial secretions of the patient post^ 



Methods 



ILung tissue removed for primary bronchogenic 

carcinoma is dissected and smears are made from selected areas 
in the bronchi:, These smears are examined by means of exfoliative 
cytology techniques o The findings are recorded with respect to 
the exact location of the origin of the smears = 

The tumor itself is hemisected end Imprint smears are made on 
slides. These imprint smears are con^ared with the positive smears 
made from bronchial secretions obtained during diagnostic workup of 
the patient for better understanding of cellular types o 



131 



COlSrriNUAIION sheet - l Serial So. WCjL.-lQI 

(contn) 

During the postoperative period the bronchial secretions of the 
patients are repeatedly examined to determine the persistence of 
malignant cells that could come from the extirpated primary or from 
second primary or metastaslSo This should give Information on whether 
a second primary Is present in the other lungo 

Patient ^laterla_l ; Adult patients on the Thoracic Surgery 
Service at the M. Do Anderson Hospital and Tumor Institute. 

Major glgdlnRS g At present this study consists of 44 cases 
The final evaluation Is awaiting the collection of additional materials 

Sig nificance to t he Progrjm_of the Inatltuteg (1) Studies of 
specimens taken from the bronchi of the resected lung should add to 
our knowledge of the routes of spread of cancers within the lungs ^ 
Any information concerning the factors involved in the dissemination 
of cancer cells should help provide a more rational basis for the 
surgical approach, (2) Ii^rints from the cut surface of these cancers 
serve as an excellent source for comparative exfoliative study of the 
several types of bronchogenic carcinomao Coniiarisons of the pre^ 
exfoliative with the exfoliative cells permit a better histologic 
evaluation of the tumor,, Thus^ the inoprinted slides are especially 
valuable for teaching purposes^ (3) Studies of the patients' sputum 
following lobectomy will contribute to the diagnosis of bronchogenic 
carcinoma in a contralateral lobe before it becomes apparent clinically 
or in radlogrsmS; and to a better understanding of the biologic behavl^}'! 
of the disease, and should have some value with respect to prognosis , 

Proposed Course of the Project g Accumulation of pertinent data 
as indicated above will continue over a number of months before 
statistical evaluation is completedo 



g,agt_B._ included Yes/ZT WJU 

132 



PES-NIH 

Individual Project Report 

Calendar Year 1959 



Serial No,, HCI-lCl 
1- Field Investigation? an 
Demonstrations Brancb 
2o DiagDOBtic Development Program 
3= Durham, North Carolina 



Part A, 



Project Title: Diagnostic studies of gastric carcinoma 
Principal Investigator: Dr. Jane G«. Elchlepp 
Other Investigator: Dr. J, C. Hijmans 

Cooperating Units: Duke University Medical Center 

Durham VA Hospital 

North Carolina Memorial Hospital 

Lincoln Hospital 
Man Years (Calendar Year 1959): 



Total : 


1 3/4 


Professional: 


3/4 


Other : 


I 



Project Description: 

Objectives : l) To assay possible changes in gastric secretions 
and tissues by chemical and immunochemical procedures to determiae 
their significance to the diagnosis and prognosis of gastric 
carcinoma o 

2} To assay possible changes in other gastrointestinal 
secretions and in blood by cheruical and iramunochemicaJ. procedures 
to determine their significance to the diagnosis and prognosis of 
trie carcinoma. 



3) To detect and evaluate possible se^iuential functional 
processes in the development of, and reaction to, malignaxit changes 
of gastric tissue. 

Methods Employed : In general, the procedures eniimerated below 
are to be used in the study of gastric secretions, blood, salivary 
secretions, intestinal secretions, and csmcerous and non-cancerous 
gastric tissues. 

1) Electrophoretic studies of proteins and glycoproteins and 
immunoelectrophoresis studies of blood group mucoids, 

2) Chemical studies of proteins, glycoproteins, hexosamine, 
fucose, sialic acid and blood group mucoids, 

3) Blood group studies of titre levels and secretor status, and 

h) Histologic studies with particular emphasis on tagged anti- 
body methods. 



133 



CONTINUATION SE'. NCI-IO8 



Patient Material; Material is obtained from patient.- ac i.ake 
University Medical Center, Lincoln Hospital, Durham VA Hospital, and 
North Carolina Memorial Hoepital« 

Major Findings: None, This project vas proposed in March 1959, 
and fosnnally organized in May 1959= Laboratory space and e>AUipment 
provisions were completed by approximately October I959, Personnel 
recruitment is not yet complete. Collection procedures and all 
analytical procedures have been tested and standardized and routine 
processing of material is in progress.. 

Significance to the Program of the Institute ; Infomation 
obtained from the project outlined may be of value in developing 
a profile of diagnostic studies which woiad be of value in the 
detection of gastric cancer. Stress is placed upon the blood group 
substances in this study because of repeated correlations noted in 
statistical studies between occurrence of blood group A and gastric 
ceircinomao 

Proposed Course of the Project ; l) Establishment of normal 
values for the laboratory procedures noted above with regard to age, 
race and sex„ 

2) Analyses of materials obtained from cases of gastric car- 
cinona and certain other selected gastrointestinal diseases „ 

S) Correlation of laboratory data with clinical history and 
coux'se o 



Part B« included Yes l~J Wo /X/ 



134 



Serial No. NCI-IQQ 

1. Field Investigations a.', 
Lviiual Frojec;;. Repor-,; Demonstrations Branch 

Calendar Year I959 2 liagnoatic Deirelopment Program 

3- New Haven, Connecticut 



Part A. 



Project Title: Fluorescent microscopy studies in neoplastic 
diseases and in conditions manifesting anti- 
nuclear factors - Itew Haven, Connecticut 

Pi-incipal Investigator: Dr. Paul Calabresi 

Other Investigator: Dr. Stuart Co Finch 

Cooperating Units: Yale University Department of Medicine 

Ifen Years (Calendar Year 1959): 



Total : 


3/4 


Professional: 


3/4 


Other : 






Project Description: 

Objectives ; The use of fluorescent labeling techniques is being 
studied in application to general cellular biology with special 
attention to the problem of improving methods for detection and 
diagnostic accuracy of neoplastic disease. This includes: 

1) The use of tetracycline and/or- other fluorescent compounds 
as a label for malignant cells in tissues and bone marrow, peripheral 
blood and other body fluids, 

2) The use of the fluorescent anti -globulin technique to attempt 
the detection of: 

a) anfci-leukocyte factors in aleukemic leiikemla, 

b) anti-nuclear and anti-platelet factors in clinical eni.i ie'^ 
•w5iich may be confused with leukemia, and of 

c) anti -tumor cell factors on the cells and in the sera of 
patients with neoplastic disorders. 

3] An attempt to further aepaxate, identify etnd specifically 
label any or all of these factoi-s ^en and if detected. 

Methods Employed : The cells are studied by fluorescent microscopy 
after in vitro histocheudcal staining with fluorescent anti-globulin 
smd related techniques or after in vivo administration of tetracycline . 
More detailed methods included in publication (see Part B. for refe.rence 

Patient Material : In- and Outpatients of Yale University Hospital o 



135 



SeriaJ. Eo, W:. 



Major Findings : An initial report (see Part Bo) has been 
published. 

Significance to the Program of the Institute : Recent literature 
has implied strongly that fluorescent techniques may well assist 
the study of malignant tissues in the cells o Should this prove true 
it is ajiticipated that this project will contribute specific infonna- 
tion useful in diagnosis of human cancer c 

H'oposed Course of the Project : It is proposed to continue 
the project through I96O. 



Part B. included Yes /x7 No fj 

136 



FHS-NIH Serial Noc NCI- 109 

Individual Project Report 
Calendar Year 1959 



Part B , Honors, Awards, and Publications 

Publications other than abstracts from tJais project: 

Calabresi, Po; Edwards, Eo A^} and Schilling, R, F=: 
Fluorescent Antiglobulin Studies in Leukopenic and Related 
Disorders o Journal of Clinical Investigation, Volo 3b, p. 2091, 
1959^ 



Collateral publications by. the principal investigator 
include: Gastric Pepsin Secretion and ABO Blood Groups in 
Polycirthemia Vera: in The American Journal of Diges'&ive Diseases, 
New Series, Vole k, no» 7, pp. 515-521, July 1959* by Sievers, 
Mo L=; ajid Calabresi, Po 

Polycythemia vera: !» Clinical and laboratory manifestations: 
by Calaljresi, Po and Meyer, 0« 0. in Annals of Internal Medicine 
(in press) « 



Honors and Awards relating to this project: 



1.-^7 



PHS-NIH 

Individual Project Report 

Calendar Yea?' 1959 



Serial Ho, JCI;'_^UO 

1. Field Investigations and 
Demonstrations Branch 
2« Diagnostic Development Prog 
3o New Haven, Connecticut 



Part Ao 



Project Titles 



Orotidyllic Acid Decarboxylase Activity in the 
Pestphesal Blood and Tissues of Patients with 
Neoplastic Disease sm.d Normals^ 



Principal Investigatorss Ds-, Paul Calabresi 

Other Investigators? Dr„ Stuart C« Finchj Difo S» Cardoso 

Goopeirattng Units? Yale Univessity School of Medicine 

Man Years (Calendar Yea? 1959) s 
Total s 1/4 
Profess ionalt 1/4 
Other s 

Project Descriptions 

Objeetiveas This study is designed to determine wiiether the 
level of this is^ortant etisyme in nucleic acid metabolism is elevated 
in cancer patients sad whether this determination has any value in 

the detection ott diagnosis of neoplastic disease. 

Methods Eaiployedg Surgical specimens, bone marrow aspirates and 
peripheral blood samples will be tested for enzvme activity measured 
in terms of millimics'omoles of orotic acid-7-C decarboxylated per 
mg. dry weights Specimens may be obtained concomitantly with material 
for fluorescent studies. 



Pafclewt Materials 



Major Findings; 



Inpatients and outpatients of Yale 
University Hospital 

Encouraging preliminary results suggest higher 
activity in neoplastic tissues as compared to 

normals., 



Significance to thp Pro^rg3[a_of trhglnstituv-e s Recent literature 
binas that one disferaace between normal and malignant human cells 
may lie in altered cytoenxjrme patternSo Hopefully this project 
could furnish a measurable parameter for identification of human 
malignancies o 



Proposed Course of the Projects 
end intensified in^WsO, 



Part B, I:;:,cluded 



Yes./ 



This project will be expanded 



No/X 



138 



PHS-'HIH 

Individual Frcjecte Seport 

Calendar Yeair 1959 



Serial Mo. MCI-lIl 

1. Field Investigations and 

Demonstrations Branch 

2. Snviroffiaeatal Field Studies 

Section 

3. Boston, Massachuaetts 



Part A. 

Frojsct Titlss Leukesaia Occurireuce in the Mew England States. 

Principal Investigators Br. Mirism D= Manning 

Other Investigators? F. A. Lawrence, B. E. Carroll 

Cooperating Uaitsi Maine, Kew Hampshire, Veamont, Connecticut, 
and Mass&chusette State Health Bepertments. 

Msi& Years (calendar yeex 1959); 
Totals i/3 
Professional: 1/3 
Others 

Project Descriptions 

Obj ectiv e; To determine the occurrence of leukesaia by tovn, and 
age group in the six Mew England States, thua giving an objective 

analysis of leukesaia in a selscted area. 

Methodas 1) Conference with State Health Officer in the selected 
states to gain their interest and cooperation in the study. 

2) Visiting the depsrtraenfc of vital statistics to get cases by 
age, residence and any other pertinent data that are available, 
such as certificate infonoatlon. 

3) Obtaining population data from states - by county, toun, and 
age 

4) Calculation and collation of rates, by town, county, and age. 

5) Comparison of data for variation frora one locale to another. 

6) Vital Statistics Departiuents in Maine, Kew Haapshire, and 
Hsssacbmsetts sre sending in copies of death certificatea of 
ell esses of leuk^oiffi as they occur. 



139 



GOcrrDJU&TlOS! SHEET 1 Serial lio- NCI-Ill 



Ma,;jor Findings; Eesults thus far indicate that there is a significant 
difference in leukemia rates for all ages during a two<>year period 
between Worcester County in the central part of Hassachusetts and 
gasex County in the northeaatera part of Massachusetts. Eates continue 
to be higher than average in Bseex County and lower than average in 
Worceetar County. Suffolk County, which contains Boston, continues to 
run a lit Lie below average. Likewise in New Hampshire > the leukeaia 
rate for all ages is significantly higher in SSerrimack County than in 
Hillsborough County. This ia interesting because the countiea ere 
adjacent to each other on the Sferrtoack River, aad Eillsboro County 
contains the industrial city of Manchester while l&rrimack contains the 
residential city ox Concord. Sxplamation will be eought for these 
facts as the study continues. This will be done by breaking down the 
figures into exact age and exact residence in each case. T^is analysis 
may fore the basis for detailed environmental studies of the areas 
concerned . 

S i^aif icance ; i) It will provide Accurate data for ieukeaia occurrence 
in otx& geographic area. 

2) Geographic areas are being located where detailed environsiental 
studies should be done in connection %rith occurrence of leukemia. 

3) These data vill halp determine rsletionship of leuktsaia to 
iamiedlate enviroiscEent . This may give soiae clue as to the etiology. 

4) The Isukesia data thus detailed are available for comparison with 
those ttc^ ocker cancer in children and adults. 

^Proposed Cooree^^^ of ^^ .^gro jec t s 1) To coagjlate collection of data on 
states already started. 

2) To initiate data collection for selected states not yet studied. 

3) To vM.ps collate, and correlate leuksaia distribution aad rates. 

4) To subject to statistical analysis the results coisplled. 

5) To coi2Sid@r possible studies of envlroreaental factors in all areas 
vhere the occurrence of leukeaia is high, in an effort to evaluate 
poeaib Is causative factors. 

6) If lasasiagfui results are obtained, to extend the study to other 
forsss of cancer. 



lun 



COSTIKUATIOH SHEET 2 Serial Ho. KCI-iil 



7) Because of the small number of deaths from leukemia reported 
each year, this project will necessarily have to continue several 
year* in order to gee statistically reliable figures. 



Part_J. Included Yes l_J Ho /X/ 



141 



PHS-NIH 

Individual Project Report 

Calendar Year 1959 



Serial No. NCI- 112 

1. Field Investigations and 

Demons cratiohs Branch 

2. BnviroiBQental Field Studies 

Section 

3. Boston, Massachusetts 



Part A. 



Project Title; Foilowup of Children Treated by X-ray for 
Lyeaphoid Hyperplasia 

Principal Investigators Or. Miriam D. Manning 

Other Investigators » B. E. Carroll, P. A. Lawrence 

Cooperating Onics° Children's Medical Center, Boston, Massachusetts 

Man Years (calendar yes? 1959) s 
Totals 2/3 
Professional g 1/3 
Others 1/3 

Project !>escriptions 

Objectives; 1) Determine whether or not children previously treated 
by XT ay in the nose and throat area or for heiaangiama in any part 
of the body are any store prone i.o develop leukeala and other 
laalignancies than chil&en not so treated. 

2) To have a series of cases \jhich will ccsapore with similar work. 
done in this country and in Bogland by other investigators. 

Methods jgaployeds 1) A letter and question sheet was developed 
vhich includes pertinent date concerning the child X-rayed, 
including siblings and psreats. 

2) This question sheet, together vlth an explanatory letter, vas 
mailed to parents of the children treated by X'^'ray. A self-addressed 
stamped envelope was Included to make answering more probable. 
Question. sheets returned were tabulated for answers to several 
questions such as? history of X«ray elsewhere, history of allergy 
in parents or siblings, and present condition of the treated child. 

Major Pindlngs ; 1) 490 children who were irradiated for lymphoid 
hyperplasia were followed for & period of at least five years. 
This represents 53 percent of the children who received irradiation. 
There was no significant difference in soorbidity or loortallty of 



11^2 



COmxm&TlOn SHEST i serial So. HCI-112 



la&lignant disease between the treated children and their 
uatreeted siblings Mortality and seorbidity rates did not 
differ sigaificantly ixtm gensral population rates In the 
same age group. 

2) Tha data obtained on a ssaall group of children in this 
study does not indict therapeutic Irradiation as a factor in 
the etiology of malignancy in childhood. The data, however, 
do not exonerate irradiation. Only a few cases of aalignancy 
in the group, not heard from, would radically change the 
conclusions. A l&sgex group with better followup is necessary 
for iBore conclusive evidence. 

Si|^aiflcance; 1) This will help to answer either in the 
affinnative or negative whether X~ray of the nose and throat 
area predisposes to leukeiaia or other tnalignanciea. 

2) This study in children can be cccapared when coc^leted with 
the study done by Court Brown in adults and also with the work 
of Slrapson and Hes^laioan in children treated by X^ray for 
enlarged thyiaus. 

3) It will answer questions which parents have concerning the 
desirability of therapeutic X-ray for their children. 

Proposed Course of Projeet g This study was ccnapleted in 1959 
and it is proposed to extend it to several thousand children 
treated with X»ray for beraangioeaa. Data are already being 

collected for this. 

Patient M aterial g Children who have been and are being treated 
for lye^hoid h3^erpl&aia and haoangioaa in the Children's 
SSsdical Center in a large northeastern city. 



Part B. Included Yes / / Bo 7x7 



143 



Individual Project Report 
Calendar Year 1959 



Serial Mo. MCI-'il3 

1. Field Investigations and 

Demonstrations Branch 

2. Environmental Field Studies 

Section 

3. Boston, Hassachusetts 



Part A . 

Project Title; 



A Study o£ Stiological Factors in Leukemia end 
Other E&lignancies of Childhood 



Priacipai Investigators JSr. Miriaa D. Hoonlng 

Other Investigators s Dx. Kaiser, B. E. Carroll, P. A. Lawrence 

Cooperating imitss Children's Medical Center» Boston^ Massacbusette 

Man Years (calendar year 1959) i 
Totals 1 

Professional; 2/3 
Other? 1/3 



project Description: 



iectlves; 



This study ccmbines material secured on all 



children with Eaalignaacy seen at Children's Medical Center in a 
large northeastern city during 1959 and the five preceding years. 

2) To determine Aether anything in the environment or medical 
history of the aother (especially during pregnancy) has oon- 
tributed to or precipitated malignancy in the child. 

3) To assessble facts on the child's environment and his medical 
history. 

Methods Employed s 1) Mothers of all children with malignancy, 
ages 0'»14, are intarvlewed early in the course of the child's 
disease, using a question sheet previously described. 

2) Hospital records are reviewed for further data. 

3> Industrial exposures of the mothers are followed where 
practicable. 

4) Hater ial is collated for cooESBon factors and the differences 
are noted. 



1^^ 



GOIfTIMJATION SHEET 1 Serial Ho. KCI-113 



5) others are separated Into three following groups for 
ccmparisons 

a. Mothers of ieukeinic children 

b. Mothers of children with other laalign&ncies 

c. Control groups 

1. Orthopedic clinic 

2. Cystic Fibrosis clinic 

6) All children seen at the Kedical Center are being skin 
tested for response to 72 caceaon allergens. The program will 
be continued until 250 to 300 have been tested. 

7) The results of skin testing will be compared for children 
with -vaxious types of malignancy and those without malignancy. 

Ma^jor Finding s; 1) 180 cases of leukemia and other malignancies 
have been added to the 517 reporLed last year. 

2) The results In preliminary tabulation bear out the increased 
incidence of allergy and therapeutic irradiation of the mother 
in children with leukemia. 

Slgntf icaneeg 1) This study has uncovered the facts noted 
above %^ich, with further studies may help establish an 
etiological factor or factors in acute leukemia in childhoods 

2) This study already implies that leukemia may have a different 
etiology from other cancers of childhood. 

3) The finding of decreased incidence of allergy in the other 
malignancy group as coeopared to the leukemia group places further 
Importance on allergy as suspect in the etiology of leukemia. 
The finding of less exposure to therapeutic X^ray correlates with 
known facts concerning the etiology of leukemia. 

4) Facts obtained have led to oiore objective methods such as 
skin testing all children with malignancy for response to coosnon 
allergens. 

5) This material can be cosapared with work being done by other 

investigators. 

Prog>o g ed Course of Project s 1) To continue interviews with 
mothers of leukemic children until statistically significant 
numbers are obtained in all groups. 

2) To tabulate results for correlation of factors in cosmon. 



lUt; 



COHTISJUAXIOM SHEBT 2 Serial Mo. NCI- 113 

3) To correlate the known significant factors with child's re- 
sponse to <a) therapy (particularly response to ACTH and 
Cortisone), (b) course of the disease, and (c) blood findings. 

4) To develop further laboratory tests which will corroborate 
the findings of the questionnaire. 

5) This study will be correlated with studies being done by 
state universities and health departieents in representative geo'^ 
graphical areas of the country. Quv aia in this study is 500 
cases of leukestia in Boston, 500 cases of other malignjmcies of 
children in Boston to coapare with double that nuadier ^ich would 
be obtained by state ualversitius and health departaoents in their 
own studies of leukemia. 

6) To extend studies to other age groups. 

Patient Materials 1) Children with leukemia and other malignancies 
seen at Children's Cancer Research Foundation in a large north- 
eastern city. 

2) Children with cystic fibrosis and normal children seen in 
the Orthopedic Clinic at Children's Medical Center in a large 
northeastern city. 



Part B, included Yes / / No /Y7 



146 



PHS-NIH 

Individuel Project Report 

Calendar Year 1959 



Serial No. BCI°ll6 

1. Field Xnvesclgetlons and 

Demonstrations Branch 

2. Snvironmental Field Studies 

Section 

3. Bethesda, Maryland 



Part A. 



Project Title's Geographic Distribution of Respiratory Cancer 
Mortality in 15 Selected States. 

Principal Investigators P. Lawrence, J. lAn. Lloyd 

Other Investigators s B. S. Carroll, S. Siwsterraan 

Cooperating Units; Rone 

!ten Years (calendar year 1959): 
Totals 1 i/3 
Professionals 1/3 
Other % 1 

Project Itescription; 

CfeJ^ctly es; To discover geographic locations of any areas within 
fifteen (15) selected states which aighL be experiencing unusual 
mortality rates due to pulmonary cancer. 

Methods Employed ^ 1) Special tabulations covering a ten<'year 
period were obtained from the Nai.lonal Office of Vital Statisclcs. 
Analysis will be made of all cancer deaths due Co oral and pulmonary 
cancer occurring smong residents of the selected states by year, 
county, race, sex, and age. The states Included ax&t West Vlrglnla» 
Virginia, Maryland, Pennsylvania, Ohio, Kentucky, and North Carolina; 
Missouri, Arkansas, and Louisiana; Arizona, Utah, Colorado, New 
Mexico, and Montana. 

2) Ccsnputations to be made include age^speclfic rates by state, 
county, site, sex, and race. Age-adjusted rates will be computed 
for inter- state and inter-regional cosapasiacfTi. Ratios of observed 
to expected cases based on state races will be used for intranstate 
comparisons by county. Standard tests for significance of differences 
will be applied in cooperation with the Bicwaetrlcs Section of National 
Cancer Institute. 

Major Findings s No major findings at this tiizte. However, some 
Interesting geogrt^hlcal differences have been notad. 



147 



CO»TIMUATIO» SHEET 1 Serial No. HCI-il6 



Significance ; With full realization of the shortcomings of 
cancer oortality data based upon death certificates, nevertheless, 
it is believed that the body of information being developed in 
this project will serve a nuoaber of useful purposes in the search 
for etiological agents in the hui&an population^ For exasiple, 
geographical patterns may evolve to suggest new leads for detailed 
field investigations and experiments designed to explain marked 
differences in observed lung cancer rates; the data are already 
serving in tests of previously announced impressions and hypotheses 
based on short-term observations; they will provide handy reference 
taaterlal as new leads are developed in experimental laboratories, 
and finally, they will be highly desirable as a baeelinc against 
which to cfmpsse future inforoatioa 

Proposed Course of Project; Computations for final analysis have 
been completed. Dusking the year a report will be prepared cover'^ 
ing the results, either for publication or for adralnistratlve use. 



Part B. Included Yea / / No /TT 



148 



PHS-KIH 

Individual Project Report 

Calendar Year 1958 



Serial tto. HCI-U ? 

1. ?ield Investigations and 

Demonstrations Branch 

2. Envlroranental ?ield Studies 

Section 

3. Bethesda, Maryland 



Part A. 



Project Titlej 



Possible Effects of Atomic Fall*out a« Reflected 
In Vital Statistics 



Principal Investigator; B. E. Carroll 

Other Investigators; P. A. Lai^rence, Dr. V S. Archer, 
Dr. S. C. iQgrahaia XI 

Cooparating Units i ^ne 

|£an Years; (calendar year 1959); 
Total: 
Professional; 
Others 

j^oject Descriptions 

Ob jec elves ; To detect possible hanaful effects of fall-out frost 
nuclear weapons testing through their influence on those vital 
statistics which would be affected by hanaful genetic changes in 
Lhe population or by increased cancer incidence. 

Methods Employed ^ 1) Tcema of vital statistics which might be 
affected by direct or by genetic effects of radiation from atomic 
fall°out have been tabulated for 'a. base-line period of 10 years 
and for each year following the start of nuclear weapons testing 
in Nevada. 

2) The items tabulated include sex ratio among live births, infant 
death rates, fetal death ratios, birth rates, death rates from 
congenital malformations, and death rates from cancer. 

3) The tabulations have been made gaographically on the basis of 
the best available estimaLes of the asount of fall-out received 

in various regions, states, and counties in the search for possible 
changes along this, path which do not occur in other areas. Charts 
have been constructed to show trends for each variable studied. 



1U9 



COirrmUATION SKBET l serial »o. NCI- 117 



Major F lndlngj?; No evidence of chsoge hes been discovered to 
date vhich could not be attributed to chance fluctuations. 

Sig;nlficsnce; 1) Any increase in cancer mortality along the 
path of atotaic fall-out which did not occur in other areas 
and not otherwise sscplained would raise the presumption of 
increased cancer incidence due to radiation damage. 

2} Any unexplained changes in the other iteias tabulated would 
raise the presumption of radiation dsausge to the populations 
concerned and of possible increases in cancer incidence s^ae 

years later. 

proposed Course of Project s 1) Data for the base°line years, 
end through 1955 have been tabulated as vital statistics 
became available for succeeding years ^ 

2) Results will be presented in reports for administrative 
use from time to time. 

3) The project will need to be continued for a few years in 
order to have a chance of producing definite results, either 
positive or negative, 

4) Routine reports of atomic fall°out data now being furnished 
by the Radiological Health Division of PHS will continue to 

be received and vie%red for appropriate comparison with our 
data. 



Part B. Included Yea I / Ho / X / 



1 tin 



PHS°NIH 

Individual Project Report 

Calendar Year 1958 



Serial No. KGl-il9 

1. Field Investigations and 

Demonstrations Branch 

2. Environmental Field Studies; 

Section 

3. Bethesda, Maryland 



Part A. 

Project Title; Geographic Distribugion of Cancer in the U. S. 

Principal Investigator; p. A. I.«wrence, W. Lloyd 

Other Investigators; 6. E* Carroll, C. Pinker ton 

Cooperating 9nits; {lone 

Man Years (calendar year 1959); 
Totals 1/3 

Professional; Less than 1/3 
Other; Less than 1/3 

Project Descriptions 

Objective s; To i^p the disurlbution of mortality due to sealignant 
dlse&se within the United States and to see if any geographic 
patterns exist ^ich might be correlated with variations in knoi^n 
geological, topographical, or other features of the country. 

^tho d8_^toalo;^ ed; (1) This project was initiated severe! years 
ago by calculation and mapping of relative frequency of cancer as a 
cause of death in each of th@ counties of the United States for a 
3 year period "•' 1949, 1950 and 1951. Raw data sources were the 
National Office of Vital Statistics and Bureau of the Census. Coisr 
parlsons aaong counties within each state revealed interesting and 
suggestive geographical patterns ^ich frequently extended across 
several state boundaries. 

(2) Eecent access to che 650 cosaputer pemitted production of sore 
accurate ^aeasures of cancer isortality over & longer time period for 
all counties in the United States. These seasures are in the form 
of mortality ratios standardized for age, race and sex, corapt^ted for 
& 5 year period centered around 1950 These data have now been 
plotted onto maps and the geographical patterns «^ich have eoerged 
are under intensive study by c^sparisons with published map» showing 



151 



COMTIHUATIOS SHEET 1 Serial No. WCI-119 



broad geophysical features of the country. 

(3) Work is ncre going forward on a further refinement of the 
ssortality ratios to take into account such factors as urbanization, 
physician density^) and proportion of foreign born population. The 
650 co^uter »ill be used for these refinements. 

j^ jjDg_Finding8 ; Froea spot maps prepared so far, it is apparent 
that human cancer occurrence and environstent, in a broad sense, 
have a saeasurable positive correlation. The most recent plot of 
all the standardized mortality ratios reveals some provocative 
geographical patterns which see@ to correlate surprisingly well with 
certain broaii geophysical features of the United States. ?or 
example, in the eastern half of the Onlted States (east of the 102nd 
Meridian) there appear two broad and different patterns of cancer 
experiences 1. Elevated cancer experience seems, to occur in the 
northeast portion of the country composed of all the Hew England 
States, New Jersey, and all of the states which border on the Great 
Lakes. (A similar pattern has been indicated in several older 
studies based on less detailed data and for different time periods.) 
This area contains approximately 48% of the l^ited States population, 
the greatest concencration of industrial activity, and the most 
highly urbanized regions of the country. Both urbanization and some 
forms of Industrialization have long bsen known to be associated 
with elevated cancer experience; however, within this broad region 
there are many counties and larger areas of low urbanization and 
industrialization which show the elevated cancer experience character* 
is tic of this broad region. In addition to the obvious regional 
characteristics, there is at least a suggestion of a possible long 
range meteorological factor provided by the fact that this region of 
high cancer experience is bounded surprisingly well by a line marking 
the maxisston southern extension of the great ice sheets which penetrated 
the Horth American continent during the last period of glaciation. 
2. Lower cancer mortality experience is revealed in a bsroad region 
ccH&possd of all the states lying south of the Ohio River and extend- 
ing westward from the Atlantic S^'^aboard to a rather sharply defined 
western boundary which runs diagonally from the southern tip of 
Texas to a point in central Missouri Within this broad pattern 
there are several anomalies composed of patches of contiguous counties 
(extending scmetimes across state boundaries) in which elevated cancer 
experience is indicated. Notable ex^nples .are to be found (a) along 
the Appalachian Highlands, (b) In the phosphate mining region of 
central Florida, and (c) in a roughly triangular patch enclosed by a 
line extending northeastward from about the center of the Coastal 
Plain of Texas, including all of Louisiana, into north central 
Mississippi and dipping southeastward across southwestern Alabssa 
to the Gulf Coast at a point in the western part of Florida. The 



152 



eCOTINUATIOlf SHEET 2 Serial Mo. MCI-1I9 

shape of this anoataly is suggestive of a prehistoric ehoreline and 
of Boms influence related to the delta-foirasissg history of the 
^ssis&ippi River. 

These t^o broad patterns of cancer experience (high and low) •>•= 
mid a few others '*'^ ae@m to show exceedingly teiapting correlations 
%7ith certain loaps sharping broad associations of Great Soils Groups 
of th@ United States. Th@ soils isaps obviously are related to long 
range effects of clisoate and westher on. ancient geological formations. 

Slgn_i f icgacg s (i) While it is not understood %dsat isisaediate 
practical significance in cancer research can be attached to these 
observations, it doss se^a issportant to ksov that real geographical 
diffes-ences in the occurrence of neoplastic disease do exist within 
a human poptjlatlcn living and dying on a land ssass as large as that 
of the United States. It is surprising and thought°provoking that 
bro«d geophysical £nd climatic features of the land mass occupied 
by humans are even s&sggested by geographic patterns of neoplastic 
disease bssed upon a time period as short as five years. Similar 
deraographic observations have proven to be of practical use in 
elucidation of the aause and control of scise other acute and chronic 
dissaees and in other research areas concerned with biological 
phencmgns in general. 

(2) In anticipation of sKodern research yielding a more crystallized 
pictiire of the mechanisms regulating the growth of living cells and 
the relation of these to neoplasia, it ^ould appear that ms^s such as 
a?e being produced iu this project could serve & useful purpose in 
tying together and possibly evaluating relative strengths of cur- 
rently fashionable concepts about the causes of "spontsoeous cancer" 
in hssaan beings. 

^opp^a9d_Coursg_of^_t^hg _gg^g;]^ec^ s (1) Further refinement of the 
cancer mortality data will be carried out and plotted. 

(2) A paper describing the results of the project to date is 
planned for publication. 

(3) As new infonsation gro«7ing out of modern research dealing with 
earth sciences beccra@3 available, it will be collated and ccsspared 
with the current cancer pattern maps. 

(4) Hew maps sho^ng cancer distribution for different time periods 
are contessplated and can be easily produced vhenever new population 
and vital statistics become available. Additional working space for 
preparation, handling, and studyof large maps would greatly facilitate 
this work. 



Part B. Included Yes J_^^/ Ko / K / 



153 



PKS-NIH 

Individual Project Itepost 

Galendsr Year 1959 



Serial Mo. «SI'120 

1. Field Investigations and 

DeiQonstrations Branch 

2. Snvirotsaental Field Studies 

Section 

3. Bethesda, Marylaad and 

Albany, Ke«r York 



Project Titles Geographical Incidence of Cancer of Major Sites 
in Upstate Hew York 

Principal Investigators B. E. Carroll ^ P. A. Lawrence 

Other Investigators s Br. S. C. Isgrahsm II, Morton LeviOj Mi Doj 
Ruth Taylor, M. D. 



Cooperating Units; Biccietry Section, HC3 
Department of Health 

Man Years (calendar year 1939); 
Total; Less than i/3 
Professional; Less than 1/3 
Other t 



Mew York State 



Project Descriptions 

Gbjectivee; 1) To study the geographical pattern of cancer 
incidence for selected sites in Upstate Hew Tork. 

2) To relate the findings to data from other studies of cancer 
incidence and ioortality. 

Methods EiB gloxgd ; i) The Hew York State Department of Health has 
made available its punch cards for reported cancer cases for the 
years 1949-1951. The data are believed to be unusually complete 
and accurate. These have been analyzed by county, city, and state-> 
wide urban'^rural groupings, using the indirect method of adjustment 
for age, sex, end urban-rural residence. A deck of 1950 New York 
population cards was purchased froia the Bureau of the Census to 
furnish the population bases for computing rates. 

Major F indinj^g; Age specific incidence rates have been cosaputed and 
age-adjustment carried out by mechanically cosputing e^ipected 
numbers of cases for each of several cancer sites for the urban and 
rural portions of each county and for each urban place. Tests of 
significance have been carried out between expected and observed 
nisabers for each site for each county, and for each city over 
10,000 population. 



154 



CONTINSATIOH SHEET 1 Sfislal No. 

/ 

Sig nificance s There is a n£ed for sy8tes&ati« c««^ilatlo<i of 
geographical variations in incidence and mortality in order to 
learn of any patterns which may emerge. Hjrpotheses as to cancer^ 
igenic factors often imply characteristic geographical distributions 
which can be verified or refuted by analyses of this kind. 

Proposed Course o£ ggojact g A geiieral paper on the study has been 
prepared with co-authorship £T&a the Hev York State Health Gepa^t" 
meat and ^I. After a long delay in the hands of the New York 
co-euthors fosr' final ccmsaents or revisions, the paper has been 
coiapleted and submitted for clearance and publication. This paper 
is titled "The Distribution of Cancer Cases in l^ban and Eural 
Areas of Hew York State", by Levin, Haenszel, Gerhardt, Ingrahsn, 
Carroll, @t al. It should b« accepted for pt^llcation ea^iy in 
1960. 

Certain detailed aspects of the data are believed worthy of 
further analysis and publication. This work will be carried on 

as time permits. 



Part B. Included Yes / / No / X / 

155 



Individual Project Report 

Calendar Yea^ 1958 



Serial No.^ Hei-l2l 

1. Field Xsvsstigations and 

Deajonstrations Branch 

2. Bnvironsaental Field Studies 

Section 

3. Bathesda, Harylaad and 

Chicago, Illinois 



MBELA' 



fjroject Titles Cancer as a Cause of ^ath aad Bisebility iraong 
Workers in the Railroad Industry. 

FriKcipal Investigator; Benjamin E. Carroll 

Other Investigators Pope A. L«wr®nce 

Cooperating %its; Railroad Retireissat Board, National Office 
of ?ital Statistics 

Msa Ysars (calendar year 1959) s 
Totals 2 
frofesaionels Less thaa 1/3 

Others 2 

Project Descriptions 

Objectives; 1) To determine occupational snd other group differ^ 
entials in mortality end disability frfm cancer of various sit«s 
(^ith particular attention to the lung) ^aong ^iployees in the 
reilroad industry. 

2) To siake cc»^arisons with figures for the general population. Any 
g isii las' i ties or differences found can serve as leads for detailed 
inveatigations as to specific etiological factors (possibly in 
cooperation with railroad laedical departxaents.) 

gfethods Employed; 1) A cohort of all railroad eaploye^s with at 
least 10 years of service, either active in 1951 or retired by the 
end of 1951, has been set up» oussbering aboat 1,000,000 .persons. 
Data for this cohort covering s&ortality fr^i all causes and disabil-^ 
ity retireaeents due to cancer have been or will ba assembled at EEB 
headquarters from punch-card files and frea clsisas. The desired 
data are being supplied to the Institute in the Form of duplicate 
punch cards, tabulations, and claiia data representing the defined 
cohort and the deaths and disabilities occUrriag aatong raesibers of the 
cohort each year. It is expected this cohort will be followed for 
3 to 5 years at least. Bata will be available to classify saeiabers 
of the cohort and the cancer cases as to age^ sex^ length of service, 
occupation and ^ployer for leaking both gross and detailed coBquctisons . 



156 



GOOTIKUATION SHEET 1 Serifti No. SCI- 121 



Since all causes of death are being accounted for, deta will be 
available of possible interest to other Institutes. The Heart 
Institute is paying part of the cost of the project, and will be 
furnished tabulstions of data of interest to them. 



\ot Findings; None as yet. 



Significances Railroad i^jployces constitute an occupationally 
stable group with their own retiresaent systesa. This is sn ideal 
setting in which to attesapt investigation of occupational factors 
and will provide good experience for later handling occupational 
data collected in Social Security Board operations. Certain 
railroad essployees are known to be e^qsosed to carcinogenic agents, 
and we h&ve here a direct zseans of observing the effects. With a 
defined cohort, holding certsin occupational factors constant, it 
should be possible to supples^nt the RES records with collection 
of additional details on personal histories (in cooperation with 
Individual railroads) to test association with other factors. 
Considerable interest has recently become manifest on the part of 
rallro&d cosstpanies and the Association of American Eailroads in 
the outcomes of this and other studies on the health of railroad 



Fro£osed ^urs c of the Pro^jeccg During the first 5 years the 
operational details of collecting the date froai BSB files have 
been worked out, the cards for the cohort have been selected and 
tabulated, 4 years* mortality and disability data have been 
c(%£plled on data sheets, and 3 years' data have been punched into 
cards. Tribulations ssre now being planned for the first 2 years' 
results (1952 and 1953). .On the basis of analysis of these data 
a preliminary paper will be prepared for publication. 



|artj|. Included Yes / / »o /x / 



FIB" KM 

Individual Project Report 

Calendar Year 1959 



Serial Ko. WCI;125 

!<. Field Invsstigations and 

Seisonstriitioas Branch 
2. SnvirozH&ental Fisid Studies 

Section 
3a Cleveland, (ftio and 

Bethesda^ Heoryland 



Part A. 



I?r eject Title; Types of Cancer Among Railroad SatgieeerSg Firemen, 
and trainnen. 

Principal Investigators s F. A. i;saw<reace, J. ffia. Lloyd 

Other Investigators; B. I. Carroll, S. Ssith 

Cooperating Units; brotherhood of Locomotive Firenen atnd Sngineaten, 
Brotherhood of Railroad Trainmen. 

Man Tears (calendo; year 1959)s 
Totals 1/2 

Professionals Less than 1/3 
Others 1/3 

Project Dsacription; 

^jlectiv ea; Phase^ls 

1) To study age and occupational patterns of cancer nortality 
asBong a particular segsaent of railroad es^ioyees. 

Fhase_^^2; 

2) To study certain detailed aspects of oalignancies causing 
deaths a&»ong a specific segEzent of railroad eaiplpyees, covering 
detailed site, s&etastatic sites, if any, histologic type, and method 
of diagnosis. 

3) To c&spaxe the findings indicated above with those obtained for 
other population groups such as those covered by the 10-city and 

Iowa surveys, or with a matched control group. 

Methods Eaiploy^d; 1) 2>ata have been obtained froa the Insurance 
Departs^nt records of the Brotherhood of Kailroad Traixaten and the 
Brotherhood of Loccmotive ?ireaasn and Engineaen covering &ge» length 
of saisbership, and local lodge affiliation of all members carrying 
life insurance, and corresponding data for all insured members 
dying frosa 1950 through 1953 These cases ere being looked up in 



1 ^P 



GOMTINUATIOH SHEET 1 Serial No. »C£°125 

death certificate fil&a of WVS to detesmina causes of death fosr 
aimlysla according to the variables indicated. 

2) Data are being obtained fron the Insurance Department records 
of the Brotherhood of Railroad Traicsiien and the Brotherhood of 
Locoaaotive Firesaen and Engineiaen covering pertinent information 
on a ctsrrent bssis on total insured messbership and on all insured 
members for whom death claisae are filed. Sach caae vill be looked 
up in death certificate files of WVB and cause of death determined. 
For each death attributed to cancer, a letter and return report are 
being sent by the Brot:herhood to the attending physician, request<~ 
ing the pertinent detailed information. 

3) Age-adjusted mortality rates for cancer (as well as for other 
causes) will be computed for each occupational group. In addition, 
patterns of occurrence of various histologic types of neoplasm will 
be analyzed in relation to the mortality rates, in addition to the 
proportions microscopically confirmed, proportions with stetastjises, 
etc. for ct^^arison with other population groups as indicated above. 

Plndlngg; None ao yet. Work is still in progress on the first 
phase and dsta are being collected on a current basis for the 
second phase, idilch began on January 1, 1956. 

Significance g 1) While this study is, in its first phase, similar 
to the lallroad letirasient Board study, its real significence lies 
in the detailed data being obtained in the forward^ going study of 
phase 2. 

2) Through the second phase Infomation can be obtained on certain 
aspects of cancer occurrence in railroad esaployees which are not 
available through the Railroad Retirement Board study. The latter 
is limited to anonymous statistics, v^ile the brotherhoods ere 
assisting us in obtaining data on each case concerning method of 
' diagnosis, histologic type of nsoplaera, etc. which will be a 
valuable supplement to the general mortality data. 

Proposed Course^ of ^ Pro^lect; 1) The next few months should see the 
completion of the £isst phase, nes^ly analysis of the data obtained 
fxcm existing records. The work has been delayed due to the work 
load on projects having higher priority, but Is now going forward 
re^idly. 

2) Since the second phase covers a limited population (approximately 
120,000), data covering 3 to 5 ya^rs will probably be needed to yield 
significant results. The lag of a few months in availability of death 
certificates at N07S will mean that the project will require 1 to 1 1/2 
years to cooiplete after the collection of basic data has been finished. 

Part B. Included Yes J^ Ho /x^? 

159 



PHS-HIH 

Individual Project Report 

Calendar Year 1959 



Serial No. MCI- 126 

1. Field Investlgatioaa and 

BemonstraCions Branch 

2. Bnvlronmental Field Studies 

Section 

3. Hagerstown, Maryland 



Part A. 

Project Title; 



Geographic aad Snvlroiaaental Factors in Cancer 
Mortality in Ifaehington County, ISaryland 



Principal Investigators; Dsr. V. B. .^cher, DTo W. Y. Chen, and 

F. A. Laurence 

Other Investigators; 3. E. Carroll, H. Greenville p C- Finkerton, 
R. Schrohenloher, Dr. R> QamaTon 

Cooperating Units s Washington County Health Department, State of 
Maryland; U. S. Geological Survey 

Han Years (calendar year 1959): 
Total; 18-i/2 
Professionals 4'=-i/2 
Others 14 

Project £)escriptioni 

Objectlves's 1) To determine geographic patterns of cancer mortal- 
ity and trends for & period of several decades in one toerican 
county. 

2) To relate geographic patterns of cancer mortality to measurable 
variations in enviromtental factors in search of clues toward the 
etiology of "spontaneous" human cancer 

3) To develop and test methods for application on an enlarged scale 
to the problem of explanation of known geographical differences in 
cancer occurrence within the United States. 

Met hods Employed; 1) House-to-house interview, collecting personal 
data on each occupant, and obtaining infonsatlon on many environ-^ 
mental factors. The interview inforstation is supplemented by 
obtaining inforaiatlon relative to all individuals who have lived in 
the house for the last ten years. This provides the basic epi- 
damlo logical data. 

2) Extraction of envirormental information frraa available maps and 
records relative to each house. This includes approximately fifty 
different items, including such Itsss as distance from roads. 



160 



COOTINUATION SHEST 1 Serial Ho. MCI' 126 



atresssSf other houses, physiography, drainage, soil tjrpes, geology, 
vegetation, etc. The sources of these Items Include geological 
maps, tax maps, highvay maps, soil maps and alrphoco maps. 

3) An airborne gaoana radioactivity survey has been transformed 
into a map of l80»radloactivity contours. This map facilitates use 
of the airborne data in estimating differences in the background 
levels between geographical areas and even residences. 

4) Detailed physical measurements of environnental factors are 
being made in areas of high and low cancer density and at selected 
residences. Staples of soil, water and vegetation are being 
analyzed for approsiatately fifty eloaects and several other 
characteristics such as organic material in soil and radon con- 
tent of water. Ground scintill^seter surveys are being made is 
these special areas and houses. 

5) A geohydrologic survey, including a plezometric map of underground 
vater supply, has been completed in one unusually interesting 
coDEBunity. 

iSany of the above deterainatlons (especially radiaaetrlc) are 
being done at the Eagerstovm laboratory. It is planned to obtain 
many of the others by ealesion spectroscopy under contract from 
some outside source. 

6) Many of the data have been coded and put on IBM punch cards for 
analysis by life table techniques. Some of the data may be unsuit°> 
able for analysis in this way and so will have to be done by hand 
or by keysort cards. 

M ajor gl n diags s 1) There are statistically significant differences 
in cancer mortality rates between several of the election districts 
in Washington County. This la based on data adjusted for age. 

2) For 7,490 single residence houses in the city of Hagerstown 
there was & greater number that had 2 or 3 cancer deaths than 
would be expected purely by chance. However, such items as age 
of houses or selective population movements could possibly 
account for these differences. 

3) A houses tchouse scintillcmster survey has confirmed our 
impressions that the airborne gms&a survey gives an excellent 
average background figure for sizable areas or groups of houses, 
but that for any analysis requiring pairing of houses or accurate 
data on individual houses it is inadequate and must be supple« 
mented by ground measurements. 



16i 



C0i3'nMUA.TI0M SHEET 2 Serial Mo. BCI-I26 



4) Tbers is extensive pollution of "he wall waters in one town 
having high cancer mortality. This vas shown by detergent . 
analysis end coliform culture done oa all veils in or near the 
town. Wate; in this coomunity also shsws a peculiarly high 
nitrate content. 

5) A preliminary geochetnical study of -ocks, soils and plants 

in Washington County seeas to point to%rird interestir^g variations 
in the occurrence of such biologically significant elessents as 
manganese, aiolybdenum, zinc, copper, chromium, titaniuss and lead. 

Si^nificBnce; Th« idea that human cancer say be related to 
transmissible or variable enviroiESisntal factors recurs per°- 
sistently in world literature and folklore. Hotions about such 
things ss '^cancer houses" and "cancer fatailJes" have enjoyed 
periodic bursts of fashionable interest yet, until recent years, 
very little actual field work seeias to have I sen done to develop 
unbiased evidence to establish the existence ^f such things. 
Washington Qountyg Maryland provides unique opportunities tp 
ass^ble facte in considerable detail about d stribution of cancer 
among a fairly stable population group, thezv., combined with 
aodern scientific au>thod8 for collectii^, and processing for 
analysis, large volumes of appropriate environiestal data afford 
new opportunities for evaluating this old ide«^. This project may 
coQt?^:bute significantly new knowledge and at; i he least provide a 
human proving ground with which to test new coacepts about etiology, 
and jaaybe even prevention, of cancer, as they caerge from the 
intensive •and far" flung bench laboratory resea'.'ches currently in 
progress . 

Proposed pours ci of Pro ^j^ectg 1) Cos^letion of the house- to,- house 
visits in all tvie election districts of tlashio^ton County. . (Sight 
are completedg 3 partially completed. Remaining are 10 rural 
districts and 6 ws-thin Hagerstown). 

2) As Index Houses (houses in which people h<.ve developed cancer 
after prolonged residence) are identified it is planned to pair 
them with houses of siniil^' age in which no r.ancers have occurred. 
The residents of these paired houses will be subjected to ouch 
more intctnsive questioning regarding their 'sabits, social and 
physical environraent. H&ny aeasureiBsnts of enviroDoiental factors 
which would be impractical on a county-°wide basis, will be made 
relative to these paired residences. 

3) Additional help and consultation from the Geological Survey. 



162 



Cmsimkrim sheet 3 Ssxlal ho. HCI<»126 



4) CoosultatioQ with & humaa geneticist regarding the "ln£er'=> 
breeding" probabilities la Washington County. Although a stable 
population is good Sox cancer studies from the residential standi 
point, this 8£ffiie stability lasy promote IntenBorriage of related 
individuals (such as 3rd or 4th cousins). It is conceivable that 
such intermarriage would affect cancer rates. 

5) Contractual axrangerasnts for eiaisalon spectroscopic analysis 
of several hundred specissens annually. 



Part B . Included Yea /Y7 No / / 



163 



Individual Project Heport 
Calsadax" Year 1959 



Serial Bo.__^I^12£ 

1. Field Xavestlgations and 

Demonstrations Branch 

2. En'^ironaientai Field Studies 

Section 

3. Salt Lake City, Utah 



Fart^^B. Soaors, Awards and Publications. 

Fufaiic&tions other thsn {abstracts frosa this projects 

Lfisrrence, F. A.; Clien, W. Y. ; A Project for Studying the 
Geographic Ulstribution of Cancar Within & Single County 
as Related te Envirosaentel Factors; American Journal of 
Fablic Health, ¥ol. 49, Ho. 5, pp. 668-674; Kay, 1959 



ISk 



PHS-MIH 

Individual Project Report 

Caiendsr Year 1959 



Serial No. J?CI»i27 

1. Field Invest igationa and 

Desjonscratiotts Br£nch 

2. gnvirorsBental Field Studies 

$«stioa 

3. Salt Lake City, Utah 



Part A. 



Frojsct Title; 



I>svelopia6nt of BIsthods for Measuring Sxteraal 
Radiation Sjq^osure in Mines as Related to 
Caocesr Occurrence of l^aniuia Miners. 



S'riacipei Investigators s 



Victor E. ixches, M. D. 

Richard Blanchaard 



aad 



Other lavastigatoxrsg Buncan Holaday* and staff 



Cooperating Iftiitss 



* Salt Lake City Occupational Health Field 
Station, OHF Frogresa, Bureau of State 
Servicss, USPHS 



HsB Year 8 s (caiendsr year 1959); 
Totals 
Professional; 

Others 



Project Descriptions 

Objectives 2 1) To ascertain ouaBtitative external radiation expoB' 

mx-T»igvtjinuu Liji;f'»ii tcmpm- . ' '^ '^' 

ure incurred by uranium sainers. 

2) To relate this ^>!;ternal radiation to the x&crbidity and mortal^ 
ity experience of uraniusi miners. 

^^tbods ^Effljg loafed g 1) Use of the "radon ch^aber" of the Salt Laka 
City Occupational Health Field Station to evaluate performance 
of our radiation detecting devices in ataiospheree containing 
kno^m amounts of radon and radon daughter products. 

2) Protection of radiation meastiriag devices from radon and 
radon daughter "plate out" by means of plastic covers that are 
to radon. 



3) Placing radiation detection devices (with proper protection) 
in mines and leaving Lhea for a predetanuined time before they 
are resioved and re@d. Simultaneous measurements of radon 
daughters using other methods will be isusde. Attempts will be 
i&ade to correlate the meaaured concentrations of radon daughters 
with external radiation dose to sniners. 



165 



^ 



COMTIHUATIOH SBSET 1 Serial Ko. !iCI»127 



Ma J or ^ Flndin^g; A trial of pocket ionization chasabers with 
various types of protection gave extremely inconsistent end 
unreliable results. Film badges gave consiateiit results when 
enclosed in £lr«-ttgb;t containera. Rubber and neopsene balloons 
(used to keep "plate out" at a distauce) as protective covers 
for the ffi'Saettfing devices proved unsuccessful because of the 
rapid penetration of radon through the wails of the balloons, 

Gs3m!& ray measuriements taken in two ventilated uranium mines 
showed a fsirly good correspondence between Eseter readings, film 
badges, and pocket ionlzatioa chaabersi in stoppered tin cens. 

It is felt that ^en there is good saine ventilation, snd ^han 
most of the gasssia rays cotae froai the mine wallSj the correlation 
will b© good bet«feen these different measuring devices. However, 
when the ventilation is poor and Bjost of the gaaaaa cosaes frosn alr^ 
borne redon daughters ^ as in the radon chssaberj then the correiatioa 
is less good. 

Sljgnificagce; lesults of this study should make a major coiitri- 
bution toward the 3olutlo:a of scsae of the practical proble^a in 
dose iae£si!r@!2@nts of enviroi^isatsl radi&tion sxposures received 
by individual miners. Such jsessurexseats axe issportsnt for 
correlation with physiological findings of the uranixaa ralner 
project. 

^;ogosQ£^^Brse^ofJProjec_ti Since neither of th© involved 
investigmtors were sble to work on this project during 1959, and 
since nsithar ar@ new st&tioned in the Colorado Plateau sresj 
this project mast be considered as t-erminated even though its 
taajor objectives have not been achieved. 



Part B. Included Yes / / Ko / 2/ 



166 



FHS-HIH 

Individual Project Reposrt 

Calendar Year 1959 



Serial Ho. IfCX»l28 

1. Field Invastigatioas and 

Bemonstrations Branch 

2. Environmffintal Field Studies 

Section 

3. Salt Laka City, Ut®h 



L^£^' 



frojsct Titl®; Study of the Caccei: Sxperience of Son-Uranium 
Hard Rock Miners in the Eocky Mountain Area 

Friscipal Investigators! ¥. E. Archer, M. D. » and M. 1. Irjavec^ a.H« 

Other Invsatig&torss P. k. Lawrenca and B. S. Csrroli, 
D. A. Holaday 

Cosperating Saltsg Certain sBiaing ccsapanies, and the Salt Lake 
City Occupational Health Field Stsfcion, OHP, 
Bute&a of States Services, FHS 

Man Years (c&lendss ysar 1959) i 
totals 1/2 
ffofsssionals 1/4 
Others 1/4 

Project Deacriptioa; 

_^jective£s 1) To evalwata the prevalence of canser of the iisng 
among underground non«urani%m miners in the Rosky Hountain area. 

2) To detersalna if a correlation axiats between the levels of 
radon in non-ureaiua raines and the occurrence of luT&g cancer, and 
othe^ causes of death. 

3) To have th^se data frosi BO>n<-»ranl,usi mines for comparison ^th 
the results of the Graniusa SUeqs' Study. 

|jgth ffld8^EiB glo;gedg 1) Aas^obly of occupational, saedical and vital 
statistics infosBiation on a composite group of hard-rock sainers 
with at least 13 years of underground working experience in Qon°- 
uraninjsa lainas in, the ^ocky gfouataia States. . 

2) Collation arad saedical statistical analysis of the collected 
date ^th particular exsphasis on lung cancer cases snd deatl%s that 

Bisy occur. 

3) lidding sneasureaidnta of r4adon in the min^a in which the study 
group worked. These measurements saust be correlated vlth ventl-> 

la'tion and projected baefosrard 30«-40 years. 



167 



Omr.imkriQn SBSET l serial No. WGI«i28 



^ajpg Findjjttgas None to date. We have collected data on a 
study group of 1766 men isiio completed a miniaiuia of 15 years 
underground at sams tizjae between 1937 and 1948. There h&ve 
bsen 370 known deaths in this group. Life tables have been 
prepared. The radon ®nd radosa daughter levels in the subject 
mines have been measured. Thay are so low that they may not 
be expected to affect the sortaltty experience of the miners , 
yet they are epprcciably higher than those experienced by the 
general population of the United States. 

Ml^MiBMIB^' ^) B^ta on bumaa baings exposed to a potentially 
carcinogenic radioactive environment will be ctKapiled and so 
bacoBse available for possible use in cancer control. At preoent, 
such information is extreeeely scarce. Iiimited studies of miciog 
cssssunities in Europe have revcalad a high incidence of lung 
cancer in mlnsrs working in m±u@s containing radioactive dust 
and g^s. Estsnaive aniiasil experiments in Europe and the jtoericas 
as well as observations made on hiaaan beings inadvertently 
ejcposed to excessive radiation over prolonged periods of time 
have established beyond doubt that ionizing radiation causes 
cancers in animals and in people. 

2) la addition, this study may yield not only information regard* 
ing levels of radiation ^ich are hartaful but also levels of 
radiation in ^ich atiners may %ork and suffer no henaful effects. 
This Information would be of vstlus in establishing recca^eadations 
for the uraniiass lEining industry. 

^Pro^aoiae d Cou r ae o f ^ ^o A^g^ ^ ^) Coiapiete statistical analysis 
of ssortali^y araong the study group is now under w&y. 

2) Coisplete "b&ck^ard sstis&ata&" of radon levels in the ssinss 
involvad. 

3) Add above inforsaation to tha present draft of isanuscript and 
present to the cejEpacies involved with a request for release to 
be published. 



?art B. Included Yes / / Ho /S / 



1B8 



PHS-KIH 

Individual Project Report 

Calendar Year 1939 



Serial »o. MCI- 129 

1. yield Investigations and 

Demonstrations Branch 
2-. Envlrozmental Field Studies 

Section 
3. Salt Lake City, Utah 



Fart A. 



Project Titles Cancer Occurrence in Uranium Miners of the 
Colorado Plateau 

Principal investigator: Victor £. i^cher, M.D. 



Other Investigators; 



Po A. Lawrence, B. S. Carroll, J. V. Lloyd, 
D. A. Holaday, Harlan Brjavec 



Cooperating Units: Salt Lake City field Station of Occupational 

Health Program; Grand Junction Operations Office 
of the Atosaic Energy Coamlsslon; the University 
of Sochester; the Indian Health Service; Lhe 
State Health Bepartsients of Utah, Colorado, New 
Kexlco, and Arizona 

Man Years (calender year 1959): 
Totals 3 
Professional; 2 
Other s 1 

Project Cescriptioas 

Objectives ; 1) To deteraine the health status of the sen vho sine 
uraalim in the Colorado Plateau ss&& of the United States, in an 
attempt co detesuine whether or not any of chelr leorbldity can be 
attributed to their occupation. 

2) To determine the frequency of different causes of laortallty, 
particularly cancer, aaong this group, coasparing them with other 
lalners and. with the general population. 

3) To determine whether biological effects attributable to radia- 
tion can be demonstrated in this group of workers. 

4) To identify any early signs of cancer and radiation injury. 

5) Atteapt to correlate morbidity and siortality with radiation 
levels. 



169 



COOTIMJATIOH SHEET 1 Sejrlai Ho. KGI»129 



^^^ thods ^^Emp loyads ' 1) Triennial physical examinations esre done on 
as taaay uranlim miners as possible wbo work in the Colorado Plateau 
area, detailed identifying dst« are obtained and a standard physical 
exmaination including chest K^x&y, blood, and urine tests is done. 
In addition, special tests which may indicate radiation effects 
are done. In 1957, this included Tised Vital Capacity, Bilobed 
Lymphocyte Count, Hair Root gxsoaination, and Sputum Cytology. 

2} An extensive follovup program, including an annual mine-to- 
iffiine vjensus, has been developed to maintain contact vith each 
aaia@r and to determine causes of death in the group. Diligent effort 
is exerted sassoog local pathologists and physicians to increase the 
rate of autopsies on deceased miners. 

3) The data collected fros the medical history and examination 
are analysed statistically to deteraine any Correlations with 
years of underground seining or the level of radiation under which 
the aainers work. 

4) Ths mortality data are analyzed by the life^table laethod and 
coctpared to other milling groups and to the general population. 

5) Frequent measurezQents of radon daughter concentrations in the 
uranium mines are made by the cooperating groups naiaed above. 

6) Searching for lung cancer deaths aiBong uranium miners in the 
past 20 years, wlio were not included in our study. This is done by? 
(1) obtaining death certificates mentioning lung cancer in males in 
selected counties of Utah and Colorado, and (2) making local inquiry 
to deteraine occupational experience of these men in uranium mines. 

Estimates of the number and age distribution of uranium miners in 
early years of the industry provide a rough population base for 
judging the overall significance of current lung cancer experience 
of the present uranitam miner population. 

Major Findings; I) Through Deceaber 1, 1959, there have been 100 
deaths of men who are included in our study group. 12 of these 
were due to malignant neoplasss (6 were lung cancer); 31 were due 
to cardio>vascular»renal disease (5 were corpulmonale); 45 were due 
to accidents (22 were associated with the men's work); 4 were due 
to suicide; 3 were due to silico^tuberculosis; 5 were due to "other 
causes," such as alcohcillsg& and infections. The median age at death 
has been 48 years. Of about SO men in our study group i^o started 
mining in 1941 or earlier, 20 have died, 4 of them of lung cancer. 



170 



I 



CONTIMUATIOH SHEET 2 Serial NOo WCI-129 



2) The spuCtaa cytology studies disclosed 14 suspicious samples out 
of 1343 performed. Although the numbers may be too sxaall for statis- 
tical analysis, they show that the percent of suspicious sanples 
increaseQ sharply with the nuaber of years of underground uranium 
mining. Furthermore « one of the 14 cases showing suspicious sputum 
in 1957 has recently died of lung cancer. 

3) A prelimiaary analysis of the hair root examinations shows that 
there are about uhree tis&es as many dysplastic hairs among uranium 
miners as ^aaong controls. Interesting observations have been made 
about the effect of age and certain chemicals on the structure of 
hair and hair roots. 

4) Freliminary results of the blood studies indicate a relatively 
high incidence of biQucleeted»lymphocyte8 in the blood of uranium 
miners when compared to blood collected from a control group of coal 
miners. They also indicate differences in lymphocytes and neutrophils 
between high and low radiation exposure groups. 

Significance s 1) During the latter part of the last century and 
early psxt of this century, death rates from lung cancer in usaniusai 
miners in Europe were in the range of 50-75% of all causes of death. 
There ia reason to believe, because of high concentrations of radio°' 
active gas and dust in mines in this country, that a similar hazard 
could exist and it is believed necessary to evaluate the possible risk. 

2) Long range study and followup of this unique industrial population 
composed of several thousand men, some of whoe are exposed to 
ionizing radiation levels high enough to produce detectable biological 
response, are expected to yield important results «^>plicable to the 
matter of permissible exposure levels for workers in other occupations 
and possibly for the general public. 

3) This study may, for the first time, doaonstrate subclinical bio<° 
logical effects in uraniian miners. These changes may be of value 

as indices of exposure to certain kinds of radiation. If found to 
be correlated with later diagnosis of cancer, these subtle changes 
might even prove useful in early detection of the disease. 

Proposed Course of Project g 1) Annual contact with the members of 
the study group for an indefinite period through census and followup. 

2) Analysis of morbidity and mortality data on the study inter- 
mittently as they are collected. 



171 



COOTIKOMIOH SHSBT 3 Serial No. MCI- 129 



3) Repeat the physical exanlnations every three yesrs, adding any 
new tests which appear to have protnise in deaonstrating early signs 
of cancer or radiation effects. The next exsesination is scheduled 
for the suomer of 1960. 

4) Complete statistical analysis of physical examination findings 
and mortality experience is now under way for preliminary reports 
on the study to be submitted for publication. 

Patient tiaterial g Efforts are laede to include ail \iraniua miners 
working in th@ Colorado Plateau Area in this study. 



Part B . Included 172^68 /—y Ho /y"/ 



Individual Frojact Report 
Calendar Year 1959 



Serial Mo. MCI"129 (aub 1) 
i. Field Investigations and 
Demonstrations Branch 

2. Environmental Field Studies 

Section 

3. Salt Lake City, Utah 



Part A. 

Project Title: Age Deteraination by Means of Hair 

Frincipal Investigators Edith Luell 

Other Investigators Victor E. Archer, M.D. 

Cooperating Units; Hous 

Hau Years (calendar year 1959); 
Total: 1/8 
Professionals 1/16 
Other? 1/16 

Project description: 

pb^ectiyeg To explore the possible correlation between age and 
hair structure and.attea^t to make it as quantitative as possible. 

Me t hods ^^ Ebjp loy e,4 ° Collection and microscopic examination of hairs 
from individuals o£ all ages. 

liajp r Findings ; In the course of microscopic examination of hairs 
from uranium minsrs and coal miner controls, Mrs. Luell discovered 
that she could observe a difference in the hair of people of dif» 
ferent ages. By looking at hairs she was often able to estimate 
the age of an Individual vithin 5 years. Cer&aln /criteria have 
been Identified which enable her to do this. 

Significance s If hair structtire is shovn to correlate well with 
age, this finding would be of importance In forensic Medicine (to 
help establish the age of unidentified bodies). It might also be 
useful in the study of the physiologic process of aging. 

Proposed Course s To analyze collected data and write a report. 
(Neither of the investigators is in a position to pursue this study 
farther, although both feel that its possibilities have not been 
exhausted.) 



Part B. Included 



Yes / / Ho /X / 



17.-^ 



Individual FX'oject Report 
Gftlendsr Year 1959 



Serial No. SCI»129 (sub 2) 

1. Field Investigstions and 

DesaoiMstrations Branch 

2. Baviroomentfil Field Studies 

Section 

3. Salt Lake City, Utah 



Part A. 

Project Titles Effect of Selenium Sulfide on Hair Roots 

FriQcip«l Investigators: Victor S. Archer, M.D., and gdith Luell 

Other Investigators; Hose 

Cooperating Units: None 

Man Years (calendar year 1959); 
Total; 1/16 
Professional; 1/32 
Others 1/32 

Project Description; 

Ob jectiyes ; 1) To determine whether or not coBsaercial preparations 
of seleniisa sulfide have an observable effect on hair roots. 

^^thod feglojed: Application of "Selsun" to the scalps of individ° 
uals followed by periodic exasaination of hair roots. 

^jor_gi nding;8 g "Selsun" has a pronounced effect on hair roots, 
very siftilar to those produced by radiation and anti<>>cancer drugs. 
This was shoim follo%fing a single application in accordance vith 
directions. 

S i j^ ni f ican c e s This finding underscores the potential toxicity of 
seleniuiB sulfide and opens the vay for the developatent of & sen* 
sitive huBian test for other toxic substances. 

Project teminated. 



Part B. Included 



Yes /X / 



«ol_/ 



17 'I 



PHS»SJiH 

Individual Project Report 

Calendar Year 1959 



:t B- HonorSj Awards and Publications 

Fublicffltions othsr ttts.n abstracts frosa this project: 

Archer, V. E. snd Lvtells I-s Effect of Selenium Sulfide 
guspensiotj. on S®ir loots. Journal of Investigative 
JSarssatoiogy. Accepted for publication but volusae iKatsber, 
page sad year not yet kaown. 



175 



PHS»NIH 

Individual Project Report 

Calender Year 1959 



Serial No. MCI- 130 

1. Field Investigations and 

Beaionatrations Branch 

2. Enviroxasental Field Studies 

Section 

3. Salt Lake City, Utah 



Part A. 



Project Titles 



Study of Occupational Backgrounds of Hospitalized 
Cases of ]Lung and Bowel Cancer, Leukemias, and 
Priiaary Mallgoant Tumors of Bone in Hales Over 
the Age of 25 Years 



Principal Investigators 
Other Investigators 



V. E. Archer, MaS. 
Erjavec, R.N. 



and Marian 



Cooperating TMits; 



; B. B. Carroll 

Salt Laks General Hospital; two Salt Lake VA 
Hospitals; LDS Hospital in Salt Lake City, and 
St. Mark's in Salt Lake City; Dee and St. 
Benedict's Hospital in Ogden; Utah Valley 
Hospital, Provo; VA Hospital in Grand Junction; 
St. Mary's and Holy Cross Hospital In Salt Lake 
City 



Han Years (calendar year 1939): 
Totals 1/4 
Professionals 1/4 
Other; 

Project Descriptions 

(felectives s 1) To determine whet occupational backgrounds are 
found in cases of the above->iaentioned sialignancies in order to 
focus attention on any occupations which by reason of frequant 
listing sight be suspected of predisposing to the specific 
nialignancy. 

2) To gather data on possible enviroisaental carcinogens. 



Methods Ed 



1) Selected interns or residents in the co- 



operating hospitals are paid four dollars for each detailed history 
they obtain frcca patients in the designated categories. 

2) Data on all the cases axa enalyzed biostatistically in search of 
factors coosaon to the personal and working histories of the patients. 



ITS 



OMSimAfim SHEET 1 Serial Mo. KCI-130 



Major Findings; No significant findings to date. Vithia the 
yesr (1939), histories were collected on 98 new caBes, bringing 
the ctsQuletive total up to 374. This body of data consists of 
159 cases of cancer of the lung; 83 leukemia, lymphoma, etc.; 
and 132 cases of cancer of the large bowel. 

Significance s It is hoped that this study wiI4 %lva useful leads 
for a more detailed study of the environmental factors in some 
cancers . 

grpposed Cours e of Pr p_.jiect; 1) Continued accrual of cases until 
larger nuabers have been collected. 

2) Biostatistical analysis. 

1) Broaden the scope of the study to include non-malignant controls 
and woisen and to include residence histories in the questionnaire. 

Pat ient Material ; Host cases of the designated cancers seen in the 
major hospitals in Utah and Western Colorado. 



Part B. Included Yes /"*7 Ho /X / 



177 



PHS-HIH 

Individual Project Report 

Calendar Year 1959 



Serial Ko. MCI" 131 

1. Field Investigations and 

Dononstrations Branch 

2. Environmental Field Studies 

Section 

3. Salt Lake City, Utah 



Part A. 



Project Title; Irradl&tioa Induced 'fumora in Children 

Principal Investigators; Dr. Victor E. Archer, and Dr. C. L. 

Sinipaon 

Other Investigators; Marian Erjavec, R.H. 

Cooperating Units; Roswell Park Mesjorial Hospital, Buffalo, New York 

Man Years; (calendar year 1959); 
Total; 1/3 

Frofessional; 1/6 
Other; 1/6 

Project Inscription: 

Ob_i_ectiyea; To calculate the risk of carcinogenssis involved in 
subjecting children to X-ray procedures. 

Ifethpda ]^ployed; 1) Collecting followup data on lj722 children 
who were given X^ray treatjsents for "enlarged thymus" in infancy. 
{3r. Sis^yson and co~workers have already cosipletad this work.) 

2) Calculation of "grasa- roentgen dose" to the thyroid, reticulo- 
endothelial, end other tissues for each child. Adding the "gr«a-> 
roentgen" doses given each child and dividing by the nusaber of tumors 
produced in tha group will give an average "tuaor Induction" dose. 
This will enable us to calculate the risk to any one infant under* 
going the procedure. 

Ma^ ^or Findin ; ^s g A hypothesis based on the sostatlc saltation theory 
of oncogenesis was developed to explain the radiation induction of 
txsaors in man. This hypothesis involves the use of dosage in gram 
red applisd to the collective tissues of a group of Individuals. 
It peradts the calculation of various tueior doses. Infants and 
children are shown to be more prone to certain radioinduced tumors 
than adults. A dose of about 4 x 10 grata rad to the reticuloeno 
dothelial tissues is calculated to produce leukemia in children 
contrasted to about 4 x 10" grem rad for adults. A solid cancer 
dose of about 1.6 x 10 grem rad for children and about 4 x 10 grass 
rad for adults was calculated. 



178 



CONTINUATION SHEET 1 Serial Ho. 



S IfffPi ^ j'ggSSiS. •^ Bvea though there are uneertalnties la the hypothesis 
aad la the calculations, the cocaervative approach to Irradiation 
of hisiaaBs, tfould «f@lcoi&e these calculations ab they enable radioloo 
gists and public health officials to saake a sesi- quantitative evalu- 
ation of the neoplastic risk Involved in irradiation of people. 

Proposed Coar ae o f Projects The work has been coi&plsted and findings 

m w iwiThiiwii ■ I n ■ Mil iiii n'^ l ia r ■ ia n <iiT i ii [■rfriiiini |— ~ ^^ 

vritten up. The paper has been cleared by appropriate HIH and SHEW 
groups. The only remaining action needed is to subait the paper for 
publication. 



Part B. Included Yes / / Ho /^/ 



179 



PHS-IHH Serial Koo NCI«92 



Individual Project Report lo Field InveBtigatioiaa and 
Calendar Year 1959 Deoonetrations fe-ancb. 

2o Epidemiology Section 

5? lleii5>hiB^ Tennessee 



Part As> 



Project Title; A study of cancer of the uterine cervix In the MengphiE* 
Shelby Ckjunty popuLatioa 

Principal Investi^tors: Mary VJalton, M<, Do, J, Williaia Lloyd, Frank E,, 

Lundin^ Jr,;, Mo Do, Winifred Mendez^ Gloria 
DaviSj Benno Ko MJJBtorey Mo Do and Alexander 
Go Gilliam, Mo Do . 

Other Investigators; Cyrus Erickson, M» Do and Bou^as Sprunt^, Mo Do 

Cooperating IJaits; llie Bep^*br!ent of Pathology of the University of 

Tennessee College of Medicine and the Meaphis-'Shel'by 
Coimty Health D^artanent 

Man Years (calendar year 1959): 

Total: 14 
Professional; 5 
Other; 11 

Rroject Description ; 

Objectives ; 

l) To evaluate nass use of the vaginal aspiration cytological test 
as a screening roethod for the detection of cancer of the uterine 
cerviic in a general population » 

S) To stui^ epideniologically the relationship "betveen carciocsBa^ 
iji'»situ and invasive cancer of the cervix, 

Ifetbods Ec^loyed ; 

Both general objectives depend m?on cytological exaaination of large 
nxaabers of \rosQn frcm the fjeneral populationj, xrlth re screening at 
approxiniately annual intervals for several years thereaftero At the 
sajue ticKj repeated nKwbidity surveys (cenvacs of all medical facilities' 
identifies ell eases of uterine cancer diagnosed in the entire populS'^ 
tion, whether occurring aaong the screened or not-screened wcaoeno 

Part B o included Yes^ Ho £J 

. 180 



COHTBIUAnolI sheet <= l Serial No. 



A pre-»screGaing laorbidity siarvey, conducted for the 2 years prior 
to the survey, provides a baseline for insasurinc the effect of 
screening on the tsubsequent level of disease in the coranuaityo 
Beginning in July 1952 cytologiciol examinations were of feredj, f^ec 
of charge^ to the entire adult female population of lfen^his='Sbclby 
Countyo As a resxilt^ vip until the closing of the laboratory over 
165^000 ucsBcn were screened in I-lenjphiSg with a little over 4o^ 
returning a year or sx>ts later for a rescresnixiG exanlnationo 'Eo.q 
cervical cancer detected at first and at later screenings permitB 
the coH5)utation of prevalence and incidence rates of in»situ and 
invasive disease in different segments of the population o The 
interplay of these rates^ pluo the disease spearing in the un-*- 
screened portion of tlie population aa detenained by morbidity surveys^ 
pemits epideniolociccl stuiSy of the relationship between the t\ro 
lesions o All taken together^ plus folloir-up observation of the 
screened^ pemit evaluation of specificity, and of the failures^, of 
the Papanicolaou test au eiaployed on a nass scale » 



Patient Material: 



■Hie adult fonale population of Menphis-Sbelby County area, 

Tbe three rasjor sources for vaginal aspiration spocinen collection 
were; (l) private physicians, (2) regular daily clinics held at a 
local hospital^ and (5) annual tecgjorary clinics held in large 
industries^ hotels^ banUSj, schools etc 



Ilajor Findings ; 



For analytical purposes, first screenings perforxaed up to July 1, 
1957 (approxiEEttcly 152>0O0) and rescrecnings xjp to llovo 1^ 1957 
, (approximately 90^000) were coded end punched, laiese include 
non-residents^ hence effective numbers for Ilemphis-Shelby County 
are accordingly smallero 

Ifeforseen difficulties have unconscionably delayed final oaalyseG 
of re suits o Present delay is occasioned by the slowness in getting 
final patlajlogy reports on suspicious cases, due to the heavy 
deparfeaental work of the collaborating pathologist <> 

Certain issportaot findings ere presently clear, however. Prior to 
the survey wHy about 30^ of ceirvical cancer cases ssaong white 
residents of frteaphis, and 155» of the non-iiAite, were diagnosed in 
relatively curable Stage I of disease » These percentages have been 
doubled for the entire ccaanunityo In addition, for those participating 
in the screening pro-am, about 85/* of cases ©f invasive disease in tte 
white and 75^ in the zson»white were detected in Stage !» 



181 



COWnMJATIOK SHEET ■= 2 Serial Ho, nCI-92 



Inccanplete tabulations now also sviggest that at least UO^ of tlic 
in-Bitu lesions cgjuaot progress to iiivasive disease since thsre 
are not enoucJi Invasive eases occurring to accoimt for the lorse 
number of in^sltu I^bIous detectedo ^e inccaaplete data presently 
available also suggest that there are age diffesrencee in the 
relationship of the two lesions ^rioich vrill have clinic^ Irngplica^ 
tionc of iiaportancco 

Slgnificamee to Cancer RGOoaj-ch ; 

Jjjipor-fcsnt inferences regarding the aatural history of in-situ and 
invasive lesions of the cervi:: v;lll be available from this studyo 

Proposed Course of Project ; 

TbB only data collected this year \rere; (l) ccai^pletion of the 
morbidity surveys thru 1958^ (2) sffiopliug of the main cytolOQr 
record file^ by Bto Lundinj, to obtain data not originally eon- 
tesiglated or codedo 

Jtos to unwlllinga«Bs of the University of Tennessee coHaborators 
to continue to hoAre our staff in residence^ future data collection 
is not planned by uso However, morbidity data oxe permanent 
records^ and eaa be collected by vis later, if the University of 
Tennessee does not do aOo 

Present plans call for renKJval of otir present small analytical 
staff frosa Ifen^ihis by July 1^. 1960« Additional analytical work 
on coded material, -vsMch are then necessary; vill be conducted 
in Silver Sprinigo 



^ 182 



Individual Project Report Serial HOv> KCI^gg 

Calen<lar Year 1959 



Part B e HonorSy Auards and Publications 

Publications other. than abstracts from this project? 

l-lpnco Hoxraverj, the first paper of a planned eearies has heen 
prepaa'^sda G::cept for nlnor revisions auoitinc final pathological 
interpretation on a fotr of the Screen I cases o Thic ia entitled ° 
"'Initial Ex feet of ConESunity-ifide Cytologicsl Screening on the 
Clinical Sta^ of Cearvical Cancer ISetecteS in An Entire Cocaaunityo 
Results in the IlGmphls-Shelhy County, (Itenncesee Stu(3y<, 1 9* 



183 



P33S»IO:h Serial Ho.o HCX-I70 



IndividTaaL Project Keport 1, Field Inveatiaatlons and 

Calendar Year 1959 Btemojastrations Branch 

2o Epidemiology Section 

5 b Betbesda, Maryland 



Part A« 



Project Title; Research in epidemiologic method^ stiniulation of 
epidemiologic research in non<»Servlce agenoies, 
and development of BOiarces of field data related 
to specific epidemiologic questions 

-PrSncipsa Investigators; Ao Go Gilliam^ M, Do^ B, Ko Milzaore, M., Do 

and Jo V/illiam Lloyd 

Other Snveetigators: 

Ckspperating .Units ; 

Mail Years (calendar year 1959)? 

Total; 2 
Profeasioaal: 1 
Others 1 

Project Pescription ; 

Objectives ; 

To examine recorded epidemiological evidence in cancer for its 
validity and to develop laore adequate epidemiological tools for 
cancer investigation. 

To stifioilate the inauguration of epidemiological studies of 
cancer ancng people and agencies having pertinent material, 
the exploitation of vhich will contrltaate to an imderBtaading 
of hiiman cancer; to provide tecixoical assistance in the design 
of est\idies asid analysis, of such material; and to explore sources 
of' material directly available to ua and ataitalsle for answering 
specific qjiestions \;hich egppear psffticialarly jaajtortanto 

Ifethods Ikig)loye d; 

Adaptation and modification of standard epidemiological methods 
already of demonstrated usefulness in cozmxunica'ble diseases 
researchj, srzd extension and application of these methods to cancer 
study; including the application of such methods to published 
data for test of va3.idity of conclusions drawn o 

^SXl^ included Yes £J No ^ 

184 



COiJTIlWAinOH SHEET - i Serial Itoo MCI^ITQ 



Hirou^ contaeta made a"b meetings of, national profousional 
as60eiati033s, lectureti to medical and public health atudentu 
^id eicploitation of persooal friendships iritb persons Sa many 
agencies, to stimulate zmd asst&t In orgSBizing studies of "basic 
epidemiologic material not directly available to the Service « 
By the 86120 meanu obtain accega to data for our ovm studyo 

lor Findings: 



■B\srther consultation vms given for a study of cancer in a large 
free-'living population among \rhich there are restrictions against 
the uae of alchoboly totacco, and coffee,, (Gilliam & Milmore) 

C^acer epideEdology lectures or seminars acre conducted at medical 
or public health schools at Ann Arbor, Mlcho, Pittsburghj, Johns 
Hopkins, UniVo of California, and at the Conmunica'ble Disease 
Center o (Gilliaa & Milmore) 

Consultation gives several investigators in the design of studies 
relating to poatiitle haaards of dia^iostic radiation o (Milmore 
& .Gilliam) 

Liaison vrith national Research Council in connection with a 
study of the relation of mustard^gats poisoning and infuenzal 
pneumonia in vJorld Mar I veterans to later developaent of lung 
cancer 6 (Milmore) 

Participated in developnent and conduct of a national cooperative 
study of leukemiac (Gilliam & Milmpre) 

Participated in IffiC conferences on design of studies in radiologists 
of health eff-jcts of radiation.. (Gilliam) 

Extension of the "Dunn model" for epidemiological study of relation;, 
if any, hetveen "ca- in- situ" and invasive cancer of the cerviXo 
(Gilliam^ Milmore, 5: Lloyd) 

PJrovision of statistical consultation and work to other Sections 
of Erancbo (Lloyd) 

Slgnificaace to Cancer Research : 

Any theories of causes of csuacer in hmnans, ftncaa ao matter what 
scientific discipline they come, will remain inadequate until 
they meet the test of consistency with epidsaniological facts „ 



185 



COinllTUATlOH SHEET - 2 Serial Ifo. IJCI-~170 



Hence, development of asore precise and powerful epidemiological 
tools is of considerable iBjportance to caacer re6ear<^o 

The future^ and "br^ad usefulness^ of research in this field 
depends partly on exploitation of material not directly available 
to \is and on reeruitaaent amd training of future workers <. 

Proposed Coigse of _ Project ; 

'So "be exploited as new opportunities arise. 



186 



PHS-IHH Serial NOo NCI^lTl 

Individual Project Report lo Field Investigations and 
Calendar Year 1959 Decoiistpations Bracch 

2 9 Epidemiology Section 
Or, Bcthesdap Maryland 



Project Title: Cancer in a closed i>opulation 

Principal Investigators; A<> G„ GiUlaais Mo D<. and Bo Ko Milmore, IL Do 

Other Investigators: 

Cooperating Units: The Vital Statistics Division of nearly every State 
Health IJepsrtoent and of several foreigji countries » 
The Tabulating Unit of NIH will key-punch the coded 
data and prepare requested tabulations « 

Man Years (calendar year 1959) s 

Totals 1/5 
Professional; 
Other: 1/5 

Project Rescript ion ; 

ObjeetiyeB; 

Primary objective is to measure risk of death from ^^ causes^ 

but Bjore particularly fi-oei cancer of th^ lung, in a closed eaiployee 

population excessively exposed to gasoline and diesel enginiF 

e:sh&ust fisneso 

ttethods Employed : 

Analysis of esqployment records of about 60^000 men eaq>loyed betireen 
1906 and 1955; determining causes of dealdx In those dead^ end ccaQ=" 
paoring death rates in this population vith that for o'^ier laen in 
the same cityo 

Major Findings ; 

Age adju8t3sent relative frequency coaajparisons sttggest that lung 
c^2cer might be excessive In this groupo Houever^ exact measures 
of risk avait computation of person">year8 at risko Most of the 
data necessary are coded, but lack of the professional aan-hours 
necessary continues to delay ccaapletion of this vorko 



Part B o included Yes [J No ^ 



187 



CQWnmkTIOU sheet - l serial Ho» MCI^lTl 



SigDlflcanee to Cancer Roseareh ; 

Sesldea paroviding infoHaation regarding risk to cancer of the 
lung^ material sJaoiiLd permit contri"butions to njstiKtdology nee^d 
in stivSying industrial group So 

Proposed Coizrse of Project ; 

Ti'acing caunes of death ia about 95^ coiaplete and has reached 
the point of uneconcanical rstiamso Causes of death have been 
coded according to the International List in use at tiiae of 
deatho .Death data to be used for computing deaths expected in 
study group^ by broad causcj have been asaenbledo The aeceseary 
rates avrait ccaiipletion of appr-oprlate poptilation estimates o 

All but about 1^000 records i^ve been transferred to IHl cards o 
Prelixsinory tab\ilations for assessing the consistency of the 
data should be cconplcted in the near future o Life table ccanputS'= 
tions irill bo laachine prograaaned by the middle of I96O0 



188 



PHS-Hia Serisg Hoo _HCZ.-173 

Individual Project Report I9 Field lavestigationa and' 
Calendar Year 1959 jDemonatrationtJ Earanch 

2o Epidemiology Section 
^0 Betbesda. Maryland 



Part Ao 



Project Title: Geograpliic^ Racial^ a&d Otlaer Factors in Distrilsution 
of Cancer 

Principal Investigators : Ao Go Gillieai, Mo D, and Benno Ko l-IHioorej Mo 

Otiier Investigators: Vlilliam J.. Pendergrast^ Mo Do, Atlaata^ Georgia 
Joba Eo Dvum^, Jr., Mo D^, Berkeley j, California 
RoTaert Lo Smith, Mo Do, Nashington^ Do Co 

Cooperating Units: 

I'Jan Years (calendar year 1959) : 

Total; 5 
Professional: 1 
Other: 2 

Project Description : 

QbjeetivGs ; 

To determinG geographic, racial, and other factors influencing 
risk to cancer,o 

Methods Employed ; 

Comparison of recorded deaths attributed to cancer of specified 
sites aaaong selected population groispso Geographic^ racial, and 
other factors arc being studiedo TSrends in cancer laortality are 
also under investigation o vVhen available^ cancer incidence data 
are also Utiliaedo 



Ma.1or Findings ; 



1) A manuscript entitled "Canparisons of mortality among the 
white and nonuhite population of the United States with special 
reference to ceaicer" was completed by Cr, Robert Lo Smithy e«ad 
suboiitted to the Journal of the National Cancer Institvttea 



Part B o included Yes £7 No [J 

189 



COlITJilUATIOI} SHEET - 1 Serial NOo HCX=.17f> 



2) A naaauscript entitled ^Tixyrotl cancer and thyrotoxicosis in 
the United States; Tlheir relation to endemic goiter vras cai^ilctcd 
by Wiliiaia J* Pender^aat, Eenno K<, Milonorc and Samuel Co Marcus 
and vas subuitted for publication o - 

Slgnifieaace to Cancer Reaeaarch ; 

The epidemiological siethod depends on tfee demonstration of differences 
in risk in people uith different cfearacteriutics, environment^ etc„ 
TbuBs the results of this projeet are descjrlptive and provide informa'^ 
tioa for use in tests of eonsietency with a variety of theories of 
cancer causatioU;, as well as leads for further exploration o. 

Proposed Course of Project ; 

Oto be continued and expasaded in similar direction with other cancers 

as personnel and facilities become availablco 

Data on the trends and geographic distribution of several cancers 
have been eoj^letedo Reports ©f these analyses will be prepared. 



190 



PHS-ITIH Serial Ifoo MGI^a75 

ladiyldual Project Report io Field Investigations and 
Calendar Year 1959 Demonbtrations Branch 

2o Epideiolology Section 
Jo Pittat^argbg Pennsylvaaia 

'art A o 

Project 1?ltle2 Epidemiology of cancer of the l^mg^ PittBlsurgh Field Study 

Principal Investigators; Co Do McClure, M, Do 5 Ao Go Gilliam, Mo Da and 

Bo Ko Milmorc, M„ Do 

Otber Investigators: '.h McDo Hoannon^ Mo Do, and Ao Ciocco, Scq Do, 

Graduate School of Public Health, Univ« of Pittsturghj 
Ko Bo Boucotg Mo Do^ Women's Medical College, 
Philadelphia, Pennsylvaniaj George Ao Shipsnan, Mo Do^ 
UoSbPoHoSo, Staten Island, Hew York 

Cooperating Units; 

Man Years (calendar year 1959); 

Total; 6 
Professionsl; k 
Other: 2 

Project Description ; 

Objectives ; 

1} To investi^te the clinical and envirosmental circumstances 
under which representative cases of cancer of the lung occur in 
a general population and to determine the characteristics which 
distinguish people who develop the disease from those who escape 
it, 

2) To partially evaluate the mass saaall film r-ray survey as a 
tool for identifying cases. 

Methods Bzgtloyed ; 

1) Personal intervie\/ of reported cases and control population 
over a period of several years, together with collection of 
augmentative clinical records froa hospitals and physicians of 
the arcao 

2) Follouup, through death certification data<^ the x«>r^ed and 
non«-x»reyed population; and special and detailed follovmp of selected 
sat£5)le of x^ray exaainees who had certain lesions, together with a 
control gro^;^ \rho irere negative to x-rayo 

'-83rt B o included Yes £J -1 q fo JW 



COMTIIJUATIOK SHEET - 1 Serial NOo NCX-I75 



Major Findings ; 



1) Hone in General stvidy, since clinical records are still "being 
co]J.ectedo 

2) Purtbsr investigation of findings previously presented revealed 
the following 5 

a) !ISse increased risk of dyiixg ftrom lung cancer anong tiie 
1955 residents who failed tp paarticlpate in the mass chest 
x-ray survey persisted during 1958o CcmpariBon of six-year 
age etandardlaed rates hetvecn survey participants and non«> 
participants ^2o\xed excessive risk csnong all 5ex»rBce cate« 
gories of the latter gro\:po Age adjusted annual 'rates show 
unifozialy greater risk eoong all non«participants except 
non«^d:dte f&aalQS} vhere the nuzabers Involved are gm«l "? and 
Bllgtit chBnesa caasc considerable variations in rates 0. 

Coinpsred to U.Sd ratcs^ tjbe lung cancer death rates Qmong 
the survey participants after 1953 are very close to UoS, 
rates while the rates innon-participants are considerably 
higher o "Kiis persistent excess risk aaong non<»participants 
can Eot be e:?plained by errors in laeldaodology or self select 
tion due to previously diagnosed lung cancers o Currently^, 
survey participation and lung cancer death rates are being 
snalyzed according to foreign birth and aconomic status of 
the .1953 residentso If a correlation is found between thsse 
parameters, soiae of the differences between the two popula- 
tions con be explained a In addition^ further elucidati(») of 
these differences m^ come with the analysis of the clinical 
and pathological data currently being collected on resident 
lung cancer deaths » 

b) A total of 6,1*56 original 70 ailliiaeter survey films 
have been re-read by Dtoctore Boucot and Shipman» Both 
readers, using the Bowcot-McClure definition of *' solitary 
pulmonary nodule," "blindly" re-read all films for solitary 
noduleSj, end other abioraialitieso To date analyses of these 
readings have revealed the following: 

1) Oh the hi^is of re'»reading relatively small randomly 
selected aaoples f^rom all categories of survey partici°' 
pants, the prevalence of solitary nodules is approximately 
2aCS^ (Boucot) or 0„6?& (Shipsaan)p This is froea six to 
. twenty tlaes the prevalence reported for the Cleveland 
Survey and frcaa nine to tweaty«nine tixaes that found by 
tl^ original readers of these survey films; 



192 



COUTIKUATIOH SHEET ^ 2 Serial HOo .KCX^ITJ 



2) In ei test of the efficacy of the Boucot»»McClure 
Sefinltionj the aolitary nodule readings of Boctors 
Boucotj Shipnan and original Buifvey readers vere coi^iaredo 
In a study involving 55I residents vith 96 per cent 
conspletenesa of follo^r-up, the S^year incidence of diagnosed 
primary pulnssnary malignancy vas 2o5 per cent ataong 
resiSents selected by the original survey reBSBTB, Co 
per cent asK)ng those selected "toy ^o Eoucot and 5 "2 per 
cent among those selected by Cr<. Shipmano 

5) The prevalence of solitary nodules in S8I original 
survey filros of persons aubseqjiently dying of primary lung 
cancer is 79 (902 per cent) using the combined blind 
re»readings of both Doctors Boucot and Shipsaano Sixty- 
three ojf 7o2 percent of the cases showed evidence that the 
solitary nodule uas the original site of the primary malignancy 

k) PrelJjalnary enalysis of x-ray data shous that appro3:i= 
mately 50 per cent of 709 survey participants who subsequently 
died of lung cancer had a definite lesion at the tine of 
survey film which could be considered mallgaonto 

"SxssG data are based on tbe cougar Ison by Bpo ShlXHaan of 
films obtained on patients at the time of diagnosis and 
the original survey filaiso Review of these films ty 
2^0 Boucot Is also plannedo 

Significenee to Cancer Reseaareh ; 

1) Provides basic information re^rdlng pathogenesis and epidemiolo^ 
of cancer of the lung which can not be determined in any other Areyo 

£1) Provides partial evaluation of inoss Esnall film x»>r8y surveys as 
a lung cancer case finding toolo 

Proposed Course of Project ; 

Contin\»2d colloetion and periodic maalyeis of followup data on 
subjects for a period of several yesa's according to objectives 
stated above 



193 



I 



PHS-KIH Serial HOo MCI^ 139 

IndiviSiial Projeet Eepoirt ii. Field Investigations aad 

CaleEdar Yeay 1959 Deaaonstratloas feraneh 

.2a Geaeral Field Stisiiles Seetlsa 
5» BQTkeleyg Calif omia. 



Part Atf 

Ps-oject Title: Relation ©f Saog to Lisitg C^aeer 

PriHeipal Isivegtigator: John Eo DunUi, Jroj Mo Bo 

Other Investigators: Vas'io\iS la^bers of the staff ©f tJje Rareau ©f 
Ciffoaic Siseases^ Calif o State Hesltix Bepto 

Coopeg'ating Units: Puads fr^ Aa^ric^ Cancer Society 

lAsm Years (ealeadas* yeas*. 1959): 
Total: 1/3 
Professional: 1/5 
Otlser: 



There is aonsider^le concern as t© the shorfe and long-'term 
effects of smogs Los Angeles lias been psrtieuLarly plagusd 
witjb. smog dtsring tfee last tea years ^ laie we3J. kaova iSTbaa 
exeess ©f lung eaacer makes tMs ^se&se one ©f speeiel interest « 
Mttough tJie HSDrtality rates for Ivsag eaaoeer in Los Angeles do 
not show any definite excess at presentj, it csuld well be tMt 
an er.cess attributable to ssiog tiioisld be s^islsed by the large 
iBsaigrffiit poptslatien that has been and is being added to that 
eityo 

ttethsds EBg>loyed ; 

Ifee plKQ of the Study is to collect popialations of Los Angeles 
residents that caa be classified as leng-termp intejnaediate^ 
and short'-tesTO residsatso Sindlar groiaping for residents of 
the San Francisco Bay Area end San Biego canbined; and for the 
rest of the State -irill be collectedo Statistical considerations 
indicate that the ssis^le eize frcis eaeh of these areas should 
be about 25^000 males 35-64 years of age^ or a total sesmle of 
75,OOOo 

The California Departaient of the PsoBTio&n Legion e^eed to 
participate in the Sttj/^o Mailings to ffiembership of over 
li}Oj,000 vers begun is Msy^ 1957o A second Jasiling to 

Part^o inelvsdea Yes ^ Ko ^ 

194 



COIJTXIIU&HOH SHEET - 1 Serial NOu 1102-139 



son^respoijSeats 'va& ears>ied out to J^e a&d a '^tirS in Septeoibero 
lo eSSltijsn to requeetiog the @ffi3ipleti^i of a q^stioxmaire by 
%hs Lsgion xoGcibcr^ a seeoz^ questloxm&ire vae eaat f®r eaos^letlGa 

■by the sspouse,, 

•Hie respozisea t©. ajelled q^testloaaaires finally provided a populatioa 
of TOgOOO jnea sasd ^O^jOOO w«saea for stucco Beeause ©f the stroag 
assoeiation of- ®20j£ljjg haMta and liasig c.aacey OGemrenee It ms^ 
■be tliat the female popiilation vrill provldb a more seasitlTe ffieo^^sye 
of ai3y effect tiiat 'may be attributafcle to SEOgo 

Setaile^ codiag of a 25 percent ssc^le d" the quegtioimalrcs item 
men ansi oodiag for dea12i seayeh bo^'e loeea eaaplete4o Beath seareli 
listings have been ccsnpletedo A ta^bulatism of the tot^l i^pulatiion 
of Biales and tiie coaipletedeDdei. saBi5>le lias laeen made to cca^sre 
tbe ■<s'Jo on age dletributioa and plaee of resldeneeo Tba saiiJle 
appears to be eatisfeetory oa these variables o 

iSse coding of qijestioiiaalres freasa vcmen for death search ha® been 
ecmpleted and listlsags have been preparedo Eetslled codisig has bees 
sarrled out on a 25 percent s@3s^>le^ as for the i^ai. 

Death feeerch of ^jese s^pulstioaa have been carried out thro\ig?i 
1958 afid 1959 deaths are eiirreatl^ being seerel^do TkstoM^ I958 
tifire have "been 72 Inog cancer deaths identified ia the male 
population end 3 ^^ng the f^oalesu 

Qaeriee @re beio^ sent to the pbysieii^is vho signed the death 
certificates to detasasine oertainty of diagnosis ead histologic 
typeo 

I'lajor FindiB^s ; 

None as yeto 

SigDJfiesnee to Ceaaeer Reseaareh ; 

To assess the possibility of sEog bs a factor in the oeeurreaea of 
Itms cancer in a h^saan population^ 

Proposed CotiTse of Project ; 

Follow assembled populations five- years „ 



195 



PES-HIH Serial Ho. mi^lkO 

iBdl-sridioal Projeet Eeport lo FieM Investigations aasdT"^^"" 

"Calejadar Year 1959 DeBBoastaratlc^s Braaeh 

So Geaeral Field Studies Seatien 
5b Berkeley^ Callforaia, 



Part Ao 



, Project fitXe: Tq ftErtfeer test tfee flodisg of as ijiereaeed risk of 
leukemia eijbseqyent to infeetios vtth cfeickea-^'pexo 

.Prlalcpal lavestigatos:: Jo&b Eo Drnm^ Jr,y- Mo Du 

Otfeer IsiTsatigator: 

Cooperstiag Unit: California State Bepartaeat of Public Health 

Mas Years (calendar jesr 1959)5 

•fBtal: 0' 



O 

Projeet Deseriptioa ; 



It was found in the case«'S©nts"ol st32^ of ehlldren deead of 
leijkeiaia^ "beisg carried <m in eollateratioa witM the groT^ kt 
• Stsnford Univereity^ that cMc]£eD«>p©x was a )sueli msre freqjaeflt 
disease iia the year parecediag leukeiaia for letfkemia shildreio 
thea' for the controls o %is was joot true for any of the other 
sbildliood fiiseasesu This ficding is to be checked by examining 
a group of chickea-pox caeea for subsequent le'SjkesDia^ usiag a - 
group 6f ffieasles cases for controls., 

Metfcods Employed ; 

Tljere are available in the California State Health Department 
case reports on 80^ OCX) chlcken-po: cases and 100,000 measles 
oases for the years 19^8-49a Punch "cards are being prepsared 
from tljese reports to provide an alphabetical listing by name^ 
8e:.j age, ifionth end year of onset, ^ad health jurisdietiono 
Leukemia death certificates for five years subsequent to these 
diseases v/ill be checked against these alphabetical listings 
to determine the leukemia mortality rates following the two 
diseaseso Th? case-control study indicates that Ghicken<=pox 
increases the risk of leukemia at least 5 times in the following 
year, and if this is t3rue the population groups are large enough 
to assure finding a significant difference o 

Pert Bo included ^^^ £7 Ko v^ 

196 



COHTSirJATIOH SHEET - 1 Serial rio„ trnj^lkQ 



It will also be of interest to learn uliether the increased 
risk of leukemia resulting from ehicken»pox, if this is true^ 
is cc.Hipeisea.tQd for lay a deficit after the first yearo Does 
cfeleken«»po:£ activate incipient disease or is the increased 
risk over and a^bove tlje norxael risk? 



Ifejor Findings ; 

Since this study was begun additional data, have some fT'om the 
ease-control study of leukemia in ebildtooodo Ttis earlier evidence 
of a strong association between chicken-pox and subsequent 
leukemia vas 'weakened by these additional datao 

The etudy of the group of chieken»pox end measles esses has 
been carried through the matching of death eertificates for • 
leukej&ia deaths for the five years subsequent to the coajsnuni- 
cable disease oceurrencee The limited smouat of information 
for matching purposes has been a major problem » Two independent 
searches for snatches have been amde, j^iose matches amMs by both 
searcStes are considered fairly reliable « Kiose made by only one 
of the searches have been reviewed by the persons doing each 
search to determine which ones could be bjrou^t to a catsmm 
agreement o "Hiere are ^9 matches that have been reached by 
independent selection and joint reconciliation,, 

Sigaifieajaee to Cancer Research ; 

In view of cvirrent interest in viral causation of esperliaental 
leijkemla in animals , the finding of a viral disease associated 
with the occurrence of leukemia in hismans would be most interestingc 

Proposed Course of Froject ; 

AccisHulated data should be reviewed and a decision made as to what 
further steps can and should be taken in identification of caseS:, 
At the mcanent there is seme excess of chicken-pox matches but not 
of the msgnitiide the earlier case-control data iadicatedo IJoa 
fxirther work is being done on this project at present because of 
the demsads of other studieso lilhen possible^ this materiatl will 
be v/ritten up as a final analysis ^id report » 



197 



PHS-NIH 
Individual Project Report Serial No,, HCI°lUl . , 
Calendar Ye^* 1959 l. Field Invcsti^tlons aod 

Demonstrations Stanch 
2o General Field Studies Section 
5o Berkeley^ California 



Bm't A„ 



Project Title; Relationship Of Cancer Ocetarenee and Soeio^economie 

Factors 

Principal Investigator: JoJm Eo Cunn^ Jr*, M, Do 

Other Isvestigator: Rayumnd Fg Kaiser^ M. Be 

Cc^perating Unit: San Francisco Health Departaaeat 

ISan Yeaars (c^eadlar year 1959); 

Itotals . 

Professional! 
Other : 

Project Etescription ; 

Objegtives; 

Evaluation of (socio «»econc8Qic factors in relation to tlse oecurreaee 
of cancer generally and specific sites, 

I-letliodii Baployed : 

1) Death certificates coded "by census tracts were available f^csa 
the San Fjrancisco Health Separttisent for the yesecs 1950»1955o 

2) Ti7o sociological studies of San Francisco were evaHable in which 
census t^@£t& Isisd ''^'^ , classified 012 three dezoensions ° Fsmily Status ^ 
Social Basik^ and Segre^tiono 

5) Sex°age»site specific death ratea for Smi PVaaciseo as a whole 
(j-erc calculated and applied to the appropriate census tract popu- 
lations ix> get expected cancer deaths by site and ©ex.. "Kiese were 
eomsjared uith observed deaths from tracts gjro\^ed according to ^se 
socio*econon4c variables being studledo 



Principal interest eentered on the Social Rank and Fondly Status 
variables o Significant correlations were found as fo2J.owss 



^rtjo ijiclujaed ■ Yes ^ No J^ 

198 



C0UTIKUA3?I0N SEESS - 1 Serial NOo NCI-l4l 



l) For males negative correlations mqtg found vith Social Rank 
and cancer of all eltes^ upper Goto tract j, atcmacla, and pancreas :, 
There vere ao poaiti'^re correlationao 

g) For feaaales Social Rank was ae^tively correlated with cancer 
of the G»Io tract ^, stoanacJi, and cervix uteri o "J&e usual positive 
aseoeiation for breast caneer was not foieado 

5} For laales Fsjaiiy Statiis showed BOiBe positive association for 
cancer of tlio prostate « 

k) For females Femily Status was positively ccrrelated with cancer 
of the stOEiach and pancreas and negatively correlated with, cancer 
of the cervix uteri <. 

§) ^e interaction of Social Hai^ sad Family Status ie evident for 
Gsaeer of the st<^aeh vhere the hi^est rates for "botli sexes was in 
t^e Lov Social Ranlc=-Hl^ Family Statue areas <> Biis is also true 
for cancer of the psacreas in fesaaleso Cancer of tbe cervts uteri 
is concentrated in the Low Social Raajk-^Low Faxaily Status areao 

Si gnifieaaee -bo Caaeer Researeb ; 

1) A nisiber of studies of socio-^econoaic factors in relation to 
c^icer have "been made—first in England^ then Densaarkj. and more 
recently in this country c Socio-econcaaic gradients have been 
found f^xly consistent for some sites; less consistent for others » 
For ejcfiniplej cancer of the breast in females does not ahosi a 
gradient in our stxidyo Baere have "been other departures from 
I^opean studies in this country and what dii&ensions best defflsn« 
etratc themo 

2) In this study three sociological din»n8ions were considered 
for sKasuring gradients » For sone sites the interaction of two 
dimeasioas appears to indicate they reinforce each other » 

Pro posed Course of Project ; 

The San Francisco data ore in manuscript form but have not been 
sutmitted for publicationo ^e Los ikigelea and Oakland data s^e 
on pxmch cards and the observed and expected nusibers of cancer 
deaths by aite^, ees.^ emd tract need to be put in the saioe card 
so tabulations can be preparedo !niis woric has bees requested 
but at a low priority because of the press of other work., It 
is hoped that the analysis of these data can be carried on in 
the near future o 



199 



PHS-NIE " 

IndivldiuEl P3Tojcct Report 

CalenSay Year 1959 



Serial 

la Field Xnveatlgations fmS 

2o Gancral Field Studleu Section 
3o BexlEeley, Calif omia 



Pe?t A« 



Project OJitles Evaluation of Death Certificate Reliability as Sotsrce 
of EnvironDKntal Cancer Inforcaationo 

Principal lavestigator:' John Eo Dunn, Jto, Mo D» 

Other Investigator; 

Cooperating Xftilt; California State Department of Public Health 

Man Years (calendar year 1959) i 



2total; 1/5 
Professional: 
Other: 

Project Description ; 



1/3 



1) To detejmine the usefulness of occupational entry on death 
certificates as a means of developins hypotheses as to increased 
cancer risks associated with specific occijpationSo 



Metiaods Employed ; 



1) ^e availability of a collection of data including detailed 
life-tiias .occi;^atlonal histories pa a group of liang cancer 
patients (most of \ihcm have since died) made it possible to study 
the agreement betireen occupational entry on death certificates 
end lifetine occvq?ational histories a Results of this analysis 
vere sufficiently encouraging to warrant pursuing such a studyo 

2) A study using this ssthod v^as established in vhich oceupa<» 
tional coding of male deaths between the sg.es of 55 and 6k for 
the years 19i^9 exxd 1951 is being doneo Coding of I950 deatha 
had already been done by the National Office of Vital Statistics 
end a duplicate deck of ptmch cordfi was saade available for this 
study o 



i Pert B . included 



Yes £7 



No^ 



20c 



COUrnJUA^IOH sheet » l .,Ser3.al HOa mi»xi-i& . 



l) Analysis of tije degs-ee of ®@?ceEK;at fcetysen qcsi^^tional 
cntsy on death certificates sid eu^tual occupatioaal history 
• abowed that a little less tdaaa half agreed on any Taasis of 
occupatioaaL characterization (uBual^o last^ or a»y five^ye©? 
occupation) a Agreeioent vith last oeei^ation was best @nd 
was in the neighlortesod of 70 per cento HSiis is fortunate 
frcsa a tecbnicalj altho^s^ not frcm aa epideiaiological^ stand* 
point in that availaMe populations "by occijpation from l^js 
Census are on that basis o 

Significsnce to Cancer Research ; 

1) It is possilale that additional increased occupational 
cgncer ritike will he identified,, Hiis will provide leads 
for further study to determine specific carcinogenic factors o 

2) Control meastjires may he indicated to reduce occupational 
ex^sure to suspected or proven carcinogens o 

3) Evaliiation of the iisefutoess of the occupational entry 
on death certificates as a readily available source of data.o 
To make recocanendations for improving 'this death certificate 
entry,, 

Proposed Course of Project ; 

"Hie analysis of these data has been canpleted aad a manuscript 
reporting the findings has beenpreparedo Most of the enalysia 
deals with socio-economic classification using occ^stlon as 
the variable for asaignsento Hhe specific oeci^ations showjsg 
excesses la deficiencieg for the various sites of cancer are 
specified. 



201 



PHS-NIH __ ___ 

[n«l3.¥ i,'l;aaJl. Prr?g- r; arid 

Calendar Year 1959 fenKjQstratiooa i5fa«Gia 

2:' General Field Studies Secti-on 

5 c. Berkeley J, California 



Part Ao 

Project Title; Occupation and Lung Cancer 

Principal Investigator; Jdba Eo Dvurm,s, Jroj, M. So 

Other IjaveatlgatorB: Various staff laembersj, Bureau of Cteronic Diseases 
California State Departaaent of Public Health 

CG©perating Unitt;; California State Departoent of Public Healtfe 

Man Years ( calendar, year 1959) ' 

Ibt&i: 1/5 
Professional; 1/5 
Other: 

Pro j gat Defeeriptlon ; 

Objectives ; 

In a prior case-control study of lung cancer patients^ in \/hiefe 
detailed occupational iiistorie© v/ere gotteuj, it was fcund tlmt, 
certain occupations appeared witla greater frequency In the lt«ig 
cancer patients ttean In the control© o Jtoong the suspect occu- 
pations were: \;el<<ferSj, various oecupaticns using asbestos in 
tlfeeir work, painters^ eooks^ marine engineers and firemenp and 
electric "bridge crane operators o Tbese findings are being 
investigated fuxiiher in the present study., 

Methods Employod s 

The present stu^ is a prospective stusSy in which popiil-atlone of 
worker© engaged in these occupations are heing asseableii to be 
followed for future lung cancer e3q)erience => Since populations 
of 5^*10^000 men 35=<3U year© of age are ^sireAj, it was necessary 
to use a mailed quiestionnaireo Besi4eg identifying infonaaticnj. 
t&e questionnaire asks for pertinent Infonsation regsxding tfce 
kind of work engaged inj, the nature of the expcsuress length 
©f time in the occupation^ and sisrakiBg Mstoryo Union orgsai- 
zations ofx'ered the best opportunity for reaching the niaabere 
deslredo Ifeey have proved to be most cooperative^, and mailing 
response has ranged between 85^90 P^r cent with three nsailiags ... 



Part Bo .^.ncludet Yes ^ No £7 



202 



C0KTIIIUAT20H SHEET = 1 Serial NOo_J£2»155__ 

"Both Nerthero and Soiathem Califcmia are ineltaded in the study o 
At present^ data collection is limited to the s©utfeem part ©f 
the state end has been csmpletedo 

In addition to the avispect occupations^ a control pepulation ©f 
utility workers has been assembledo 

Lung cancer^ ether cancer (and possibly other eauses of mortality) 
^fill be determined in these groups by checking all cancer deaths 
for the State against these assembled popiil&tionsu 

Qti^ries are being sent to the appropriate p^iysicians for diagnostic 
confirmation ©f all lung caneer deaths o 

SigDJficence to Cancer Research g 

Occtjpatlonb found to carry increased iwng (or other ccuacer) rlsK 
will nee4 to be studied for idestificatiea of the specific cause o. 
Frc5B this could coaae specific control measure So 

Ma.1or Findi ngs; 

Death clearance was coiapleted throtigh 1958 and a preliminary 
analysis made of the datao "niere have been 159 lung cancer deaths 
th&x have occurred diirtag the period covered in the analysis,, 
Several occupations appear to be associated with an excess of 
lung cancero However^ the magnitude of the excess is not great - 
being in the range of lo5 to 2 times that experienced by the male 
population geaerallyo One of the major problems in this eaad ax^y 
other study searching for variables associated with Itaig cancer 
occurrence is the masking effect of such a powerful and nearly 
universal factor as cigarette sjnoktngo It is possible that 
excesses of the magHltude found mi^t represent factors that votxld 
increase the liajg cancer risk five-fold if it co\ild be Examined 
In the absence of siaokingo 

Proposed Course of Project s 

T&e finad. analysis will not be undertaken until five years of 
follow-up death clearance has been con^leted for each of the 
occupational ^oups being studiedc. 



203 



PHS«N1H Serial NPo.JlCI-135 

lai^yldsial Project Report 
Calendes- Year 1959 



Part. Bo Bonorsj Avards^ enA Publications » 

FiAblicationa otbcr thsai abstracts frvm. this projects 

ThmUg, Jo Ep^ Jro,, Lindexij George j, Sreslou^ Lesters Cancey nsortuLity 
experience of laen in certain occupations in Califox^iao S\jSsBiitted 
for publication in ths Journal 6f the Aiieriean PulJlic Hea3.th AbsoCo 

Iloagliaj IrfXGiax* \hs Linden^ George j, Breslow^, Lester and Duanj Joto Eu 
Jro ; This xise of a mailed qjuiestioimaiye for cpideiaioloeicaA study » 
Sutmitteii for publication in Pablic Health Reports « 



20i| 



PHS-^KIH 

Itidividual Project Report 

Calendar Year 1959 



Serial Noc 



NCI=.144 



lo Field Investigations and 
Demonstrations Branch 
2„ Diagnostic Developmetrrc trogram 
3o Bethesda, Maryland 



Part Ao 



Project Title; Clinical Trials Unit 

Principal Investigator; Andrew C„ Peacock Pho D„ 

Other Investigators; None 

Cooperating Units: Clinical Pathology Department, CC 

Man Years (Calendar Year 1959); 
Total ; 1 
Professionals 1 
Other ; 

Project Description; 

Objectives; To develop tests capable of aiding in the detection 
of cancers of all sitas in early stages of development. The over=all 
aim is to dsvolcp one or more cancer diagnostic tests that will be 
simplej inexpensive, easily duplicated, and therefore^ applicable to 
large population areas on a mass basis o 

Me thod s ^nsgloy ed ; Btsic research is en^loyed upon chemical and 
immunological principles underlying tests described in the past, and 
principles which aid in the development of new tests. During the past 
years the principles upon which some general tests for diagnosis have 
been based have been evaluatedo Modification of these principles has, 
been attempted j and new procedures have been adopted which may aid in 
the formulation of new tests. Procedures developed are evaluated upon 
the serum from known cancer patients ^ and from normal people. 

Patient Mate rial; Patients and staff of the Clinical Center, 

Major Findings; 

•' " "AeceTeraf ors of Blood Cjjagulation ; Reliable methods for the 
determination of Factor V (proaccelerin, plasma accelerator globulin) , 
and for Stuart==Prower factor have beeti dsvissd. Both methods use 
purified fractions from normal plasma, and have the advantage over 
previous methods that the reagents are reliable and well defined. These 
fractions are widely available, xfhereas reagents prepared from the 



20, 



CONTINUATION SHEET ° 1 Serial Mo. NCI- 144 

Major Findings; (cont„) 

plasma of patients with presinned unique clotting defects are noto 
We have not yet started screening patient material. 

Ascites Carcinoma Cells ; (Krebs K=2 mouse ascites tumor) 
Factors affecting the size and volume distribution of ascites cel^.y 
have besn studied by the use of an electronic counter. Cell size was 
constant as the tumor grew. Several agents caused changes in the normal 
pattern; among these were X-rays and growth hormone, both of which caused 
abnormally large cells. The cells proved remarkably resistant to changes 
in the osmotic strength of the medium. In spite of the fact that these 
cells are rich in many enzymes, the ascitic fluid was found to be, for 
the most part, devoid of appreciable quantities of the enzymes. Among 
the ensymes found in high concentration within the cell 5, but not in the 
fluid (or blood plasma) were succinic dehydrogenase, glucose=6"phosphate 
dehydrogenase, and DPNH diaphorase. 

It was found that certain tetrazolium salts were reduced by cells 
without the addition of substrates. The ascitic fluid also had this 

property. 

Normal human serum was found to behave in the same way, and sixty= 
one human serums from a variety of disease states were studied to 
determine whether this property was of diagnostic significanceo 
Several serums were found to be very reactive in this test, but there 
was no obvious correlation with diagnosis. The serums from cancer 
patients were not in any sense unusual. This work was discontinued. 

We observed that ascites cells suspended in normal human serum 
clui^ed within a very short time. If the cells were centrifuged out of 
the human serum and resuspended in ascitic fluid, the cells once more 
resuspended freely. We devised several approaches to study this property s 
none of which were entirely satisfactory. As an approximate method, a 
standard suspension of cells in serum was placed in a capillary tube 
and the asiount of settling in a fifteen minute period measured. A wide 
variation in the behavior of human serums was noted; serums from cancer 
patients were distributed in the range of other specimens. Because of 
the coBiplexities of this problem and because the results were not en<^ 
couraging from a diagnostic point of view, these experiments were 
terminated. 

Krebs K=2 ascites cells, as well as other tumors have been reported 
to contain a substance called "toxohormone" which on injection into 
normal mice causes a drop in the liver catalase. We have confirmed 
this, and are trying to confirm reports in the Japanese literature 
that this, or a similar substance may be found in the urine of cancer 
patients. This investigation has just started, and at present no 
definitive results are available. 



20l^ 



CONTINUATION SHEET - 2 Serial No, NCI- 144 



Signific ance to the P goggam of the In stitute; The devalopaant 
of any test that would aid in the detection of early cancer in one ox 
more sites would be of great value to the clinician. Hith strong 
Suggestive evidence at hand that the patient has a cancer, the present 
diagnostic procedures could be intensified in studying this patient; 
and there would be impetus for the development of further diagnostic 
procedures c It is evident that the development of such tests wou d be 
of great practical value in combating the cancer problemc 

Proposed Course of the Project; f resent plans call for a concerted 
attack on the tosohormone question.. We intend to study this material .ts. 
the ceils J-^'i '^'^'■^ ascitic fluid , and the urine of patients with and with<=- 
out cancer The many excellent studies to date have not clarified the 
chemical nature of toKohoriaone definitely ^ and we hope to make contri- 
butions in this regard. We hope to find, eventually^ and adequate 
demonstration of the material in urine. 



Part B included Yes/"Xy No/~/ 



207 



PBS-NIH 
Individual Project RepoEt Serial No. KCI°I44 

.Calendar Year 1959 



Part B, Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Williams, G„ Zo and Peacock,A, CoS Ultra Violet Studies of 
Tetrazolium Reduction in Living Cells, Exptlo Cell Research, 
Supplement 7 (in press) „ 



Honors and Awards relating to this project; 
None 



208 



Serial Eo. 



NCI°145 



PKS-NIH 

Individual Project Heport 

Calendar Year 1959 



1, Field Investigations and 

Demonstrations Branch 

2, Diagnostic Development Program 

3, Boston, Massachusetts 



Part A. 



Project Title: A study of lymphonia cases treated in a selected 
hospital in New England, including observations 
on patients emd records 

Principal Investigator: DTc John Bo Vander 

Other Investigator: Dr. B. Ko Milmore 

Coopei-ating Units: None 

Man Years (Calendar Year 1959): 
Total ■ - 



Professional 
Other 



Project Description: 



A sxirvey of environmental and clinical factors 
common to patients with lymphoma 



Objectives : To obtain data relative to environmental, medical 
personal, and familial factors that may be associated to a greater 
degree than normal for patients developing lymphoma. It is designed 
to serve as a corollary study to that being carried on with children 
at another hospital » In this case, however, an adult population will 
be used but with the same emphasis on history of exposure to chemicals 
and history of allergy. 

Methods Employed ; Preliminary phases of the study will consist 
of two parts as follows: 

1) Review of records for the past two years of patients admitted 
to the hospital with lymphoma, for the purpose of determining 
factors which may be commonly associated with this disease „ 

2) The study of patients currently being treated, with additional 
emphasis on environmental and occupational histories, to 
elicit factors that may have a common basis in this group o 

Patient Ijaterial ; All ijatients with lymphoma admitted to a large 
teaching hospital (approximeitely k^O beds, in a New England area). 

Major Findings : Preliminary analysis indicates that records are 
very complete, diagnostic facilities are excellent, and diagnoses almos": 
invariably are confirmed by tissue study., Except for these findings, 
there are no findings in regard to the study since it is just now 
developing. 



209 



CONTimJATION SHEET - 1 Serial No<. NCI- 145 



Significance to the Program of the Institute : With the 
exception of x-radiation, and with the exception of vague report.? 
on benzol exposure, environmental factors which are common to groups 
of patients afflicted, with lymphoma are not known o Such data are oi 
importance in any attempt to elicit the etiologies of this disease, 
since it is recognized that any information on causative factors has 
a marked practical effect in preventive measures which may he taken, 
and since no curative therapy is available c. 

Proposed Course of the Project : The study will be developed 
along plans outlined, and any specific leads will be followed upo 
It is not possible at this moment to predict in what direction the 
major emphasis will be placed, since the study is in reality a 
broad investigation aimed at determining specific fields of endeavor 
that might be profitable for further study » 

Current Status ; This project continues with special emphasis 
on the treatment of chronic lymphatic levikemia with corticosteroids r, 
This phase of the project was siimmarized in a presentation at the 
7th International Cancer Congress, London, England, and has been 
further summarized in a manuscript which is presently being con- 
sidered for publication by one of the medical journals » 



Part Bo included Yes £7 Wo 7^ 

2i0 



PHS-NIH 

Individual Project Repoit 

Calendar Year 1959 



Serial No. ^^_ NCI - Ikb^ 

1 Field Investigations and 
Demonstrations Branch 
2, Diagnostic Development Program 
3» Boston, Massachusetts 



Part A. 



Project Title: Cancer incidence in a population of 5,000 persons 
examined periodically 

Principal Investigator: Dr« John B. Vander 

Other Investigator: DTc Be K» Milmore 

Cooperating Units: Kationeil Heart Institute 



Man Years (Calendar Year 1959) 
Total 

Professional: 
Other 



Project Description: 

Objectives : l) To determine the incidence of cancer of majo;*" 
sites in a selected population under continuing and regular medical 
observation o 

2) To inquire into possible etiologic factors of any unusual 
increase or decrease of cancer incidence (as compared to general 
population data) in the study population. 

Methods Employed : l) The population numbering about 5>000 
persons has been receiving biennial physical examinations with 
emphasis on detection of early signs of heart disease « The present 
project represents an intensification of eniphasis on search for 
•cancers that are accessible to direct examinationo Such cancers 
are known to represent more than 50 j6 of all cancers. 

2) There is, of course, prompt followups on individuals of 
the study group found at examination to have symptoms or signs of 
less accessible disease o 

Patient Material : The source is 5,000 presumably healthy 
people living in a New England city. This is the same group which 
is being examined periodically by the National Heart Institute o 

Major Findings ; A group of approximately 100 solitary nodules 
of the thyroid gland have been discovered, of ^ich lOj^ have been 
removed by surgery c The remainder will be kept uinder observation 
to determine subsequent changes o This study group has been kept 
under surveillance since 1955- 



21: 



CORTINUATIOK SHEET - 1 Serial No. NCI -146 



Significance to the Program of the Institute : l) Makes 
available more data on occvirrence of cancer in a selected, •well 
delineated group of statisticaJ.ly workable size. 

2) In particular will provide information about the frequency 
of maJ-ignancy in solitary nodules of the thyroid glando 

3) This well doctimented, forward-looking study is expected to 
furnish a needed test of the usefulness of occupational, history 
data in predicting possible environmental influences on occurrence 
of cancer o 

Proposed Coixrse of the Project ; 1) In line wi.th previously 
established procedures, one-half of the population group will be 
examined aimuallyo 

2) Any ceincer discovered will be treated and the patients will 
be marked down for particular fo.llowupc 

Current Status ; This work continues in cooperation with other 
members of the Framingham Heart Prograjn.. Two additions to this 
project "Which are presently being considered are routine serum 
enzymes determinations in cooperation with members of the staff of 
Brandeis University and also routine examinations of peripheral 
blood for tumor cells. 



■p'Rct Bo incliirieri Voe / / wrN ,/y7 

21^ 



PHS-NIE 

Individual Project Report 

Calendar Yeas 1959 



Serial No. NCI- 147 

1. Field lavestigatioQB and 
Desaonstrations Branch 
2<, Diagnostic Development Program 
3, Boston, Massachusetts 



Part A. 



Project Title; Cancer of bireast as related to lactation, pregnancies 
and hosstone therapy 

Principal Investigators Dr, John B„ Vander and Dr„ Rita M. Kelley 

Other Investigators? Dr, B„ K, Milmore 

Cooperating Uaitsg None 

Man Years (Calendar Year 1959)? 
Total J k 
Professionals k 
Other s 

Project Descriptions 

Objectives; (1) To attesspt to ansv/er the questidn of vhethe? 
or not hormonal treatmeat of cancer of the breast prolongs life, 

(2) To study the relationship of lactatioB; use of estrogens, 
nursing of off spiring, number of pregnancies and marital 
status to carcinoma of the breast, 

(3) To study the question; Is there a conaiKjn denominator i% 
male and female carcinoma of the breast? 

Methods EsiH>loyed g (1) This study in its preliminary phase 
consists of plans for review of records for the past five->yeae period 
with the purpose of collecting data pertinent to the above points. 
Records are those from a large New England Teaching hospital of 
anjproximately 450 beds. 

(2) Additional data will be obtained by interviews and followuv 
of patients s.v found to be necessary. 

Patient Mategia lg The initial part of the study will use the; 
recor¥s^ of Selected" patients from a New England hospital who havF. had 

breast cancer. It is planned that these patients and future breast 

cancer patie&£s at this hospital will be interviewed as necess?.iry and 

followed for as long a period as necessary to achieve the objective of 
the study. 



213 



CONTINUATION SHEET - 1 Se?i«l No. NCI- 147 



M ajor FiE dings; (I) At the present time there is no oirganized 
scientific evidence that horiaonal tireaCmeat of breast cancer prolongs 
life. If significanS; data can be obtained relative to this it would 
aid in deciding the proper place of hormones in therapy o Should they 
be used prophylactically in an attescpt to prolong life or reserved fo^ 
symptomatic treatment as the indications arise, 

(2) There is some reason to believe that the use of estrogens to 
suppress lactation may be related to the development of breast cancer 

Signif ie anee to the Program of th e Insti tute; Data as outlined 
above' might coatr ibutesignif rcantly to the pToblenj of the best hoifmoo^^^l 
control of cancer as well as to prevention, especially in regajcd to th.e 
promiscuous use of estrogenSa 

Proposed Course of the Projects in the large teaching hospital 
wh3re the records will be esamine3""is a very active breast cancer 
clinic as well as thyi?oid clinic. At the present time the principal 
investigator is engaged in doing active clinical work in both these 
clinics and is fasailias' with the patients and records,. 

The study, it is anticipated, will have three divisions? 

(1) Study of survival times of patients «ho received 
hormonal therapy versus those who did not, 

(2) Cois^erison of cancer rates in patients who have a history 
of use of estrogens before the development of breast cancer with 
those i^ho did not« This will also apply to lactations, nursing 
and marital status, 

(3) Study of common factors in male and female patients 
with breast cancer.. 

Cuyreat Statue ; Work on tMs project continues in the form of 
clinical care in chemotherapy of patients having cancer of the breast 
The data which is being accumulated will form a part of the large 
cooperative study ^Ich is being sponsored by the National Cancer 
Institute and probably will not be published separatelyo 



Part B. included 



2114 



PHS..IIIU Serial Hoo MCI^l^S 

Indiviauea Project Kepsrt. 1., Field Ii-vcstigatiooi; aad 
Calender Year 1959 Deajonstrations Braacia 

go General Field Studie* SeetioB 

and Biemetry Section 
3o Boston., Ilassachueetts 



Part. Ao 



Project Title: Cancer liortality smong Catholic Brottoers and Sisters 
in Several a?ea&hing Orders la the United States 

Pyiiicipai Inveatigfttors; B<, Ec Carroll^ and J.,. 'Jo Lloyd 

Otlier Inyestigatorss Bo K„ Miliaorep Mo Do an4 William Haeaszel 

Cooperating Units: University of North Carolina (Institute for Reaearch 
an4 Social Science )» 

Man Years (calendar year 1959) s 

fetal s 1/5 
Professionals 
Other; l/5 



Objectives s 

1) 'So determine the mortality In some of the larger Catholic 
teaching orders for all causes and particularly mortality 
attrihuted to specific cancer sites « 

2) To CQBipare the rji.sk of dying of cancer of specific sites 
with the corresponding risk in the general population to determine 
whether significant excesses or deficits exist f©y ^ay cancer aite^ 

Methods Employed ; 

l) laforoAtion on approximately 70^,000 individuals coaipriaing 
6g Catholic religious eooaaMnities from I9OO to 195^ has teen 
gathered, coded and analyzed 'by i&eans of IBA punch cards in 
another study carried oat in the Institute for Research and 
Social Science of the University of North Carolinau Althoiigh 
the fact of death was recorded and punehedj, no attempt was 
made to determine cause o 



i' Part B o included Yea fj No ^ 

i 

215 



CONT-nWAI'lOM SHEET - 1 Serial IIOc KCl=i4e 



2) Tta'OTogh 0- contract \iith the above'-najaed Icstitufee tl»e 
identifying data on all deceased individtjala teas been dupli- 
cated and a decSs Ci IHI cards on tbe entire population ijnder 
study bss "been ruraiahedo 

5} The cB^oscs of death are bejjog obtained throu^ appropriate 
chaonela i^©sn the doath certificate of each deceaaed persono 

h) Appropriate tabiilation and biocjetric analysia is to be nsade 
©f the Ejojrtality dats. to discover siiaiisrities to ftad differenees 
frcaa similar data relative to the generail p©ptilation» 

or Findings I 

Konec. Data are nov/ being collectedo 

ficance to Cancer Research 1 



This study will CQtmplejaent a similar research project recently 

coagpleted by fe'o T^lor^ Mr^ Carroll, sind M^<, Lloyd* It is wueh 
larger in scope and should help clarify saass fyagmentary evidence: 
indicating that incidence of certain forms ©f cancer la different 
for members of scaje religious order» than the general population^ 
possibly &MS3 to difference in manner of livingo Xt msy inake 
possible a significant contribution to our knowledge of cencero 

Proposed Course of Project ; 

1) Identifying information for all members of the study group has 
"been receivedo 

2} CaiiseB of death fear nearly all deceased laembers have been obtained, 
axisi this aapect of the uork utli be coa^leted early in I96O0 

3) Data on causes of death vill be put into the appropriate punch 
cards of the deck already on hand,. 

h) Llfe-'table anaJ^sls wHl be made^ using a ccaa^uter program already 
prepared for the previous study o 

5) Analysis end report are expected t© be cosiqpleted late in I96O0 



216 



PHS-KIH 

Individual Project Repos-t 

Calesidar Year 1959 



Serial SSoo ^ jgC I^- J-GS 

1. Field Investigations and 

Deinonst^aCioBS Bicsnch 

2. Diagnostic Development Frograai 
3a Boston, Massachuaetts 



Part Ao 



Project Title? Ultraviolet Microscopy as sn Aid in Detecting 
Malignant Disease c 

Principal Investigators John Bo Vaader, M„ D„ 

Other Investigator ss De, Ro F, Ssisas, DTo S, Co Ingraham II, 
Ds„ A. W. Hill 



Cooperating Units; Pondville Hospital, Walpole^ MasSo 

Man Years (Calendar Year 1959): 
Total I 3/4 
Professional; 1/4 
Other s 1/2 

Project Description; 

Objectives ; (1) To develop a sEeans of differentiating leukemic 
from non^leuksmic marrows. 

(2) To establish a method for identifying "stem cell" leukemics 
as to the e^act cell of origin„ 

(3) To ioore easily discover and identify metastatic tumor cells 
within the raarroWo 

Methojis EiEployedg (1) Bone Mar-rrov Studies. Multiple bone ■m&i^zcf: 
specisaenSj froaa patients with various types of esjalignsnt disease, tjIII 
be prepared hy direct staining with fluorescent dyeSo These will be 
studied directly under the ultrs^^^'iolet microscopeo 

(2) Esasaieation of Peripheral Blood Smears and Buffy Coats. 
These will also be escaiained by means of direct ultraviolet microscopy 
following staining with various fluorescent dyes with the purpose cf 
facilitating the discovery, and possibly identification of metastatic 
tumor cells in the circulating blood. 

This will also provide a further opportunity for attesapting to 
differentiate between the various types of leukemic cellSo Especial 
esaphasis will be placed on changes ia morphology which Eiay tales place 
under therapy o 



217 



"OFi'TIMIA.TXOK SHEET =1 Seri,; -_MirMS-- 

(coat.) 

(3) Fluorescent Labeling of Antigen. It is proposed to esplojre 
the possibility of preparing a fluorescent labeled antigen usiiig 
leukemic white cells or their products as an antigeno The immediate 
object would be the use of this substance in the differential diaguo&i!* 
of the leukejalas, especially in those instances where the question ie 
raised of the presence of such closely allied conditions as myeloid 
metaplasia oz leukeiooid reaction. 

The present plan calls for the use of white calls, or their 
extracts, farom leukemic patients as a means of attes^ting to produce 
more or less specific antibodies in eKperimental aniraalSc These 
antibodies will be fluok-escently labeled and used on bone masrow or 
peripheral blood specimens as a diagnostic test. 

More specifically white cell products will be injected into 
experimental animals c The globulin f //action of the serum from these 
animals will be subsequently obtained, treated with fluorescent 
labelling substances and tested for specificity using prepared smea;,'s 
of the original leukemic cellSo Non-specificity of antibody response, 
it is anticipated, will be 2. raajor problemo 

gat lent Material ; la^Patients of Pondville 

Major _F_ladingsj Hone to date. Project was activated in 
December 1959. 

SigiPtiflcance to the Program of the Institufces Ultraviola.;; 
Microscopy has been reported as helpful in uterine cancer diagnostic 
cytology in human ©eiags« Sons zaarrow aspirations &n& circulatir.g 
blood in leukemic patients can provide specimens physically similar 
in cell distribution to cytology specimens from cancer of other sits^s. 
It appears very worthwhile to investigate possible usefulness of 
fluorescence techniques in developing diagnostic studies activities. 

Propos ed, Course of^ the Project ; Patient data and control 
examinee data nil! be collected on blood and bone marrow speciv.nsT5 
examinations using ultraviolet fluoresceEce microscopy over the nexfc 
several years,, ^en reliable data is collected in sufficient ascnoutit 
to justify statistical evaluations ^propriate collations and analyeif 
will be performed^ 



Part Bo included ¥a s/~" / Ko^ 

218 



PHS-KIH 

Individtial Project Report 

Ceilendar Year 1959 



Serial No. NCI-l^Q 

lo Field Investigations and 
Demonstrations Branch 
2. Diagnostic Development Program 
3« Durham, North Carolina 



-t k-. 



Project Title: Hyperseiisitivity and c'ttier host tissue faetorg in 
cancer of t&e stomaclii and large bowel 

Principal Investigators: Dr. Jane Go Elchlepp 

Other Investigator's: Dto J. Co Hijmans 

Cooperating Units: D^ike University Medical Center 
Lincola Hospital 
Durham VA Hospital 
Niarih Carolina Memorial HospiteQ. 

Man Years (Calendar Year 1959): 



Total 
Professional 

Oaer 



1 3/4 
1 



Project Description: 

Objectives : l) To determine environmental emd/or genetic 
factors of the host id.th possible significance in diagnosis of 
cancer of t^e stoniach and large 'bowel = 

2) To specifically study host tissue hypersensitivity factors 
in relation to racial and sexual differences in occurrence of 
cancer of the stomach and large bowel o 

Methods Employed : l) Selection of appropriate negro and 
\?li.ite cancer patients from a relatively stable geographic and 
economic population for intensive study of the patient and his 
family with particular regard for blood group and hypersensitivity 
factors o 

2) Histologic review of surgical pathology and autopsy 
material for possible racial, sexual, or other individual differences 
in host tissues especially with regard to host connective tissue 
reactions o 

3) Immunochemical studies of blood group polysaccharides in 

the gastrointestizial tract of cancerous and non-cancerous individuals : 



219 



CONTINUATION SHEET - 1 Serial No. KCI-lJi-^ 



Patient Material ; Cases selected for this study are limited 
to idiite and negro residents of Nci'tk Cajt-olina and certain contig- 
uous coimties in Virginia and South Carolina » Patients are drawn 
from the registers of Duke University Medical Center, Lincoln 
Hospital, Durham VA Hospital, and North Carolina Memorial Hospital. 

Major Findings ; None» 

This project was reorganized early in 1959 to stress the 
chemical and iramunochemical aspects for a program of diagnostic 
studies and transferred to the Diagnostic Development Section = 
Collection of materials has continued and it is expected that 
this case study approach will be a natural concomitajit of the 
diagnostic study project. 

Significance to the Progrem of the Institute ; Statistical 
studies have reported significant racial, sexuaJL and geographic 
differences in occurrence of cancer of the stomach and the large 
bot-jelo Correlations have been noted between blood group type and 
certain gastrointestinal diseases, including stomach cancer. It 
is felt that constitutional factors in the host other than specific 
parenchymal cell suscejjtibility may be operative in the develop- 
ment and covirse of carcinoioao Intensive studies of gastrointestinal 
carcinoma in ■vdiite ajid negro patients may provide useful informa- 
tion with regard to diagnosis and prognosis of cemcer of the stomach 
and large bowel « 

Proposed Course of the Project ; This project has been terminated 
as a separate ujiit and studies will be continued in conjunction ■with 
the Diagnostic Studies of Gastric Carcinoma Project » 



Part B» included Yes [J No /x/ 

220 



Individual Project Report lo Field lave stlgat ions and 

Calendar Year 1959 Deiaonstrationi; Branch 

2. ConeraJL Field Studies Section 

iJo I5s»ry University^ GeoS*gi8 



Part Ao 



Project Title t Oral cancer in relation to vitamin deficiency, use of 
snuff 3, and other factors 

Principal Investigator: William Ho Vogler^, M„ Do 

Other Investigators 2 Jo 'Jo Lloyd and Bo K, MilMore, M„ Do 

Cooperating Unit? Emory University School of Medicine 

Man Yeax's ( calendar year 1959)? 

Total', 1 
Professional: 1/5 
Others g/3 

Project Description s 

Otjectives; 

1) To determine irhether a relationship exists Toetween the use of 
tolsasco^ avitaminosi&j, ill-fitting deaturesj, and other factors^ 
and the development of epidemjoid careinoBja in the oral cavityo 

Itethods Eaiplpyed ; 

1) A questionnaire hoii been coopleted on eH new patients seen 
in the Rotert VJinship Clinic hetueen January I956 and July 1957° 
In adfJitioUj, all old patients with mouth cancer and/or leukoplakia, 
were incltsdedo About 2000 questionnaires have been cooipletedc, 

'She infonaation aa the questionnaire has been coded and puaohed 
on IH4 cards D Tab\ilations have been preparedc 

2) All of the records of mouth cancer patients have been reviewed 
in the hope of finding 6c«ae cosanon denominator „ 

5) Appropriate statistical tests have been applied to determine 
the significance of differences between disease ^oups. 

Patient Material ; 

Patients at Robert Winship Clinic in Atlsaitao 



JBo iaeluJed Yes /^ No ^ 

221 



COIJTIflUATIOK SHEET - 1 Serial NOo KCl»i50 



1) Cancer of the laouth. is seen more frequently in waaen here 
thaffl in other ports ef the countryo 

2) It appears that the consuaapticm of eheviag te'bacco and sauff 
is much higher eDoong the caneer patients than eiaong the controls o 

Sifflii fieance to Cancer R esearch; 

Ttte investigation^ &s far as it proceeded^ hints at a specified 
relationship hetween use of snuff iand=^r avitaminoeis and the 
occurrence of cancer of the upper gastrointestinal tracto 

Propoeed Co\Br'se of Project s 

Collection of additional infonaation fnom patients has been eii,scon<=° 
tinuedc Bata alrea^ collected ara t»ing analyzedo AMitional 
tabulations will "be naeesgary« Although the principal investigator 
is ao longer with HCl^ he ig participating in analysis of the 

findings and preparation of a final report » 

Ahcut 90^ of the tabulated data and significance test coc^utations 
have "been sent to the principal investigator for prel3Mnary analysis. 
55xe remainlns coorputstions should he finished during the first quearter 
of 1960„ 



222 



PHS-NiH 

Individual Project Report 

Calendar Year 1959 



Serial No. NCI-1:)2 

1. Field Investigations and 
Demonstrations Branch 
2o Diagnostic Development Program 
3. Framinghaira, Massachusetts 



Part A. 



Project Titles Correlation of thyroid disease and breast cancer 

Principal Investigators Dr, John Vender 

Other Investigatorss Dr. R„ F„ Kaiser and DTc Bo Ko Milnsore 

Cooperating Unitss None 

Man Years (Calendar Year 1959) 
Total s 1/4 
Professionals 1/4 
Other ? 

Project Description? 

Objjctiws g To determine whether there is a relationship between 
carcincaa of the breast and thyroid disease,, 

Me th o d s _Emp lo?/ed s In a very large thyroid clinic to determine the 
incidence of breast carcinoma among the thyroid patients with special 
ee^hasis on those with myxedema. 

Patient Material; Ic Patients seen in a large thyroid clinic 
in a New England city„ 

2. Patients seen in a hospital devoted to the diagnosis and 
treatment of cancer ^ 

3c Patients seen in a health survey of a population of 28,000 
in a Hew Suglaad Ccmounity. 

Major gjadings g Kone, The project was initiated late in 1955c 

Sifflifieaa ce to the Prograia of the Institute? A number of 
observers hold the clinical izj^rsssion that- cancer of the breast is 

frequently associated with thyroid disease. This ssems to be parti-^ 
cularly true of patients with non^toxic thyroid nodules and spontaneous 
saysedema. If a relationship between thyroid disease and carcinoma 
of the breast is indeed shown to exist it would throw nsore light on 
the relationship between hormones and cancer. 



22J 



GONTIHUATION SHEET - 1 Serial No.. NCI- 152 



Proposed Coars e of the Project g . A study of patieats and rscords 
in a large thysold clinic in ^ich the ps-lnclpal investigator is doing 
clinics.! ■^jork. This clinic has bean in existence for over 35 years 
and has a very excellent, prolonged followup system possibly unsurpassed 
by any other o The records are also eKeellent both as relates to thyroid 
disease as well as other illnesaeso There is a close working relation- 
ship between the thyroid clinic and th^ breast cancer clinic in the 
ssiae instifcutiono If the suspected relationship is verified^ a more basi 
study on endocrines and cancer (with preliminary emphasis on TSH) may 
be indicated^ 

Current Stratus g This project is inactive at the present time. 



Part B, included Yes/"^ ^aJTl 



224 



PHS-IJIH Serial lIo„ MCX-153 



Individual Project Report lo Fiel4 Investigations and 
Calendar Year 1959 Dejaonatratlons Branch 

2o General Field Studies Section 

3o lOaniias City,, Missouri 



Pert A„ 



I^^ject ^I^tle;; Cancer" mortality in several selected population groi^qps 
in Ueu England 

Principal Investigatoysz Ruth. Taylor^ Mo Do^, Benjamin E<, Carroll and 

Jo William Lloyd 

Other Investig&torj Bo Ko Milmore^ Mo Do 

Cooperating Units; 

MsB Years (calendar year 1959); 

Ttetsls l"l/3 
Prof e ssionad. ; 2/ j3 
Other; 2/3 

Projeet "DcBCription ; 

Objectives ; 

1) EvBliiation of cancer mortality and morbidity in selected 
population ^oups living carefully supervised and regulated 
lives o 

2) Coniparison of cancer rates in the Btu<3y population with those 
In the general ^pulatiouo 

Methods Employed ; 

1) Kaues and identifying data uere obtained on members ccmiprising 
the study groi^ (aggregating approximately 5000 persons) and perti" 
nent date were tebulatedo 

2) Vital records of the as)propriate eoinniunttieg vrere searched 
for death certificates on deceased members of the g^o^o The 
study includes individuals dying as early as IS75 &Qd as late 
as 195^0 

; Part B o incliaded Yes ^ No £J 

i 



225 



C0IITIJIUATI0I3 SHEET - 1 Serica.HOo HCJ^JS-S 

i 



;;) Lcperted deatiis aad popiil&tion ostim&tea for tw© ccuorts ©J 
Ilasisaehusotts TaaaleB ("bom lC70«lCC9 and 10^°1909) vere oTstatncd 
f^'OD pu2:lit>hcii dat&o 

4) Calculations ©f death, ratee for the titudy ©roup and a contr©! 
population have boon iiaele ty life table teshniqueso 

flajor Findings ; 

%c stui^y bab "been con^leted anS a paper publishedo 

Evidence has "been found pointing to Important differenees in tlje 
eancer and other oorfcality of single v^o maxried women^ of nuns 
vso other single women j, and of earlier vso later cohorts of \ronen„ 
Such differences were denionstrated for total laortality from all 
causes and for cancer of aeveral sites^ tubercialosis, cardio-' 
vssct;ilar»ronal di&ease^ and all other eauseco 

Significance to Cancer Research ; 

Considerable inforoation is available covering cancer oecurrense 
In the general population and many occupational groups o Little 
is knoun about cancer in the s^ipervised and regulated population 
^©upsj certain of i4»lch arc typified lay this sty^ ©:"oupo The 
sanparison made possible "by thia gtudy oay mcH help uncover 
items in cancer etiology previoiiiily obsciired by the lack of data 
on stich population sub^oups and the paucity of valid statistical 
studies of thouo 



22G 



PHS-inCH Serial K^o_KCI«3s5X- 

Individual Project Report 
Caleadsir Yeex- 1959 

Part B » Eontarti, Av:ar4sy and Publicatioea 

Ptablicatione otbey tiian abiitaracts frtm t&ls project: 

Taylor^ Ruth So^ Carroll^ Beajaaaln E.^ sad Llsyi, Jc ivrillism; 
Mortality anssng ucaaca ia 3 CstJiolic religious ordere., witSa special 
refereacc to cancer p Cancer 12:1207^1£25 CHov<.s.DeCo) 1959" 



227 



PHS-KIH Serial Woo HCX^I54 

Iiidivi<ausl ^Project Report Ic. Field Inveatigatlons. ajad 

Calendar Year 1959 Bosiondtyatlons Bareach 

£o Gcnersl Field Stvuiiea Section 
>o ' Lsrn Asgeles, California 



Part A« 



Project Title; Excess Lvsxg Cancer asBong Mexican Females 

Principal lavestigatorB: John Eo Duiaa^ Jro, Ma Bo, Lester Griaspocn, Mc Co 

and MariB Eo Heller ^ M„ A„ 

Other lavtBstigators : Staff Ifeaaberu of Bureau of Chronic Diseasegj State 
of Calif o Depto of Public: Health; Wo M., Haeaszel^ 
national Cancer Institute 

Cooperating Units: 

Maa Years (calendar year 19^9): 

Total: 2/'j> 
I^ofesslonsl; 15/ 
Other: l/5 

Project De script ion ; 

Objectives ; 

A stwSiy of limg«»censGr asortality among Me?;ic'8« feaaal.es in California 
shovred that Msxic«M3»'boTO fereales bs-fe a threefold lung caacer ri'ak 
as coorpaxed to, other CtLlifomia wcsfaano Ho isGreased risk was found 
for Mesiaaa Jisales or Calif omia^-'bDm we^ea of Mexican eKtraction. 
Kje primciry objective of this st^l(33^■ is to dsteraxlne, if poesiMcj, 
wbat factor or factors laay be responsible for tbe ijscressed risk 
to this difsease aosong these people^ 

Cultiiiral euathropologists familiar wltli l-fexioan sultyre bare expressed 
the opinion that this excess is probably not the result of tobacco 
Usep but iQore likely^ the re stilt of the iise "of unvented ^wves for 
cooking in Ifexico, Tot^ccc and culinary feabits are tvOo but by no 
means all, of the factors with -»rtiicb this stiii^ is concernedo 

Metfcods Saployed ; 

l) To obtain inforsaation about Mexican females dying of cancer f^rcsB 
cllaical records, the ia^igration folders (through the Ju4;tiee Depto) 
and other so^srce6 of infonaatieai. Specifically desired 'are the piejje 
of birth and/or origin^ length of residence^ oeei^ational history^ 

Peart B« incliaded Yes £J Ko ^ 

228 



COlWmJNSiaS sheet - l serial HOo KCX-15^ 



yeaar of iEanigsration^ siso.Jcisag tetita^ experience witli methods ox 
cookiiig^ etCo A list of IO7 Mcixicaa^bom featales^ who mbyq too'.m 
to have died of liaifi caocerj was eubaitted to the Justice Depsartment 
in the fall of I9560 Tiale departaaent was givea 'Uie naaie ©nd ldenti<= 
fying information fo? each patient and was requssted to cwH frcea 
the patient's iaraigpatioa folder certain of the above •laentioned clatao 

2) "So obtain similar information on the Mexican female population 
of CalifomiSo 

So Sexaplc of Mexiesn Families: 

At present it is tbovieht that a eample of 1^000 Meslcaa fajssilieE 
should oomprlBe the study group., Contract with the Census Bureau 
will proviije the sample, and & contract for interviewing will 'be 
arranged with one of the several possiTsilities for prcvicling 
•brained Spanish speaking interviewer a, . Prelto»insry 41seussioas 
with STo Ralph Seals ©f the DepajrtuKint of Anthi^pology have 
been held \;ith regard to the possibility of having this inter'=» 
view perfomsed hy qualified graduate studeats In the Bepartmeat 
of Anthropology «. 

bo Ssznple of ^lexican Female Patients: 

Esaentially the saoe intearview fona that was jsaade up for the 
sample of Mexican families is joow being used to obtain Itifor-'' 
mation from female patients thirty<»f ive years or Esore of age 
who are of Mexieen origin,. All the subjects in this part of 
the study are patients on the wards of the Los Angeles County 
General Hospitals SMey are being Intervieired at a rate of 
about twenty»five a week by five or six well qualified 
Spanish-speaking interviewers on a part«tlae basis o ISiesa 
interviewers^ with one exceptioaj, were selected frcsa people 
who work in the hospital^ doisg work which ordinarily Involves 
interviewing for the Social Service 23epas-bffleat= Tlie later =■ 
viewers are being closely superlf'ised Bsad each coi^leted 
interview is being scrutiaiaed by Mrso Maria Heller , Inter- 
viewing got under way on September 12» 1957 sad el-ata hati 
been obtained on approximately 5OO Mexican fasailieso 

Patient I-teterial ; 

The patients interviewed in this atiiiSy are all patients in tke 
Los Angeles County General Hospitalo Tnsy S3:re- aH feaaalcs of 
Mexican origin &ak thirty*five years or Bssre of agej many of 
them are non-Eaglish speaking o 



229 



coimmjimm sheet- - a Seriso. ifoo kci»i5U 



tjajor Findings s 

Tlie one pacrt of tlae project whlcb has "beeri ccaapleted at tMs tlsie 
is tJsat part ^rtiicii isvolved ctiLliag certaijj inforu^tieii f^fsm tJie 
tnsaigratiOB r©14fir£ of the 107 Ifexiesa ucusen wto died of lung 
caaeero However j, eo fs\i folders uere located;, end the infonaation 
in tbosG availaJsle \rsiB eo inccai^lcte that little or no sipiificanee 
ean be given to the data that was ottaiaedo FxErthejrmore, specif i^ 
cally regarding otir interest in the state of "birth of theisc 107 
Mexican femesles^ .the Justice Bepto does not have recci'ds from vhich 
we eoiild oTjtain the yearly relative rates of endgratjon to the UoS^^ 
psartictilarly California, fraa tfee various Mexicsn states over the 
years during which thege 107 people iuEaigratcdo Therefore, even if 
ire had obtained data on the state of origin or birth on a significant 
number of these 107 Mexican females_j, the data woxjld be meaningless 
without Isnowingj for the years concerned., the extent to which each 
Mexican state contributed to the Califoroia iamigrant Ifexicaa 
population o 

No new interviews have been carried out since Efe'o Grinepoon left 
the project in Jiiue^ 1958<, Itote thsn 7OO questionnaires had been 
c<ar5)letGd at that tijoe.. A prelSainary analysis of a point of laajor 
interested was made concerning the cigarette saoking practices of 
Mexican torn Mexican woseea versus those UoSo bonao Contrary to 
the opinion of the anthropologists who ha<l tosen consultedj it was 
not found that Mexican bom wcas^n ssoked cigarettes less and 
adopted the hebit at en older age tjaaa U^So bora wc^kuo 

Significance to Cancer Reeeareh ; 

Ihe elucidation of factors associated with the developEtent of cancer 
may lead to a better understanding of the etiologf of this disease 
and, eventually J, to methods for control or prsveatioa of caacero 

Proposed Course of Project ; 

Analysis of already collected questionsairee will be coapletedo 
When this has bees accossplished, a decision will be asde as to 
whether additional questionnaires should be sc cumulated,, 



230 



PHS-IilH. Serial NOo,..,,_Ji2|riSI„ -~ -^ 

Indivi,duail Projoet Report 1„ Field IsvestigalioQs sad 
Caieadar Year 1959 DeKiOnsti»at5.caia Braacli 

2= General Field Stu<iS.es Sectioa 
5o Lob Angela b^ Cali/oraia- 



Part Aa 



Project Title: Aeljlorl^ySria Survey of tl3e Jopsaeee Pop^alatios of 

Los Angele» "by the Tufceless Gastric Analysis Ifetfe©d= 

Principal Investigator: Lester Grin spoon, M» Do^ Charles Carpester^ Bl„ D„ 

aod John E» Dunn^ Jr<>^ Mo D» 

Other Investigatoys: tfcinTaers of the Medical Staff of the Japanese Hospital 

Cooperating Units: 

Man Years (calendar year 1959): 

Ototalj a-^l/j 
Professionals 1 
Other: l»i/5 

Projeet Description ; 

Objeetivea ; 

Stcjaaach'-eaaeer mortality r&tQs macng Japssese of the United States 
and Hawaii axe saore than 5 times as ®reat as sajong the wMte popu- 
lation of the United State So Also estahlished is the fact that s, 
relationship exists tetween achloz'hyds'ia sosd gastric esascer in 
CaiacasianSo Whether a sladJ-ar relationship eziats 'between achlor- 
hydria and gastric caneer in Japanese is isot knotmo !Hie isaaediate 
goal of the project is to determine the frequency of achlorhy&'is 
among the Japanese people of Los itegeles^ Calif*, as comp^-ed to 
Mexiceua aad Caucasian control groups „ Tiie ultimate objective is 
to expand this project into a gastric caascer control progrsm "by 
doiag upper GI series on the anacid patients « IMs wou3.d enalsle 
ua to (l) teat the assianption that a relationship between scMor>»' 
hydria and gsatric cancer also exists in Japanese j (2) test the 
tubeleas gastric analysis method as a screening device with regard 
to gastric eeacer aad perhaps other diseases (cogo peraieious eseffila); 
(5) pick tap cases of asyjsiptQOjatic gastrie eoacer which caa then "be 
studied fir^a the point of view of the efficacy of early gyrglcal 
intervention o The Japasjese people coast itute sn ideal groi:^ for 
such a study in that the population is eas3J.y defined and it has 
such a high risk to gastric cancero 

Part B , included Yes ^ Ho £J 



23x 



COl^m-IUM^OK SHESr r I Sexissi. HOo HCX*157 



Methods j^ loyed; 

T&e aeid^produsiias ability of the ©tcsEScb is being Satexiaiiied "by 
tfee tubeless aaetbod, utilizing B.%M^a--^A ioa"~esQ;lsange yesiaao BlooS 
type viH also be SaScertaiBedo 

Ibe teat is, baiag pcrformeiS on private in-» aad out-patients of 
physicifiaxs on tfee staff of tJae Japanese Esspitalo Oiat^patients 
^;bo find It more convealent may report to a private leaboratery 
i» a ■business distriet on a Epecified clay each weekc. Also^ to 
sugjaent the control group^ scaae patients at the Los Azigeles 
County General Hospital ©re being testedc Ife'lne speciaens of ©11 
paarticipaaats ere analysed in the Istoratcry of tbe Japanese hospital.:. 

Physicians are notified of the result of the test,, VIBieas the result 
is "indertesmlas.te'' or ^no free ECi present^ , they are asked to 
urge tile patient t© have the rest repeated^ VJ'itb the Los Angeles 
Owuaty Hospital patients^ tbe test is repeated v/heaevar possible 
regardless of tiae reaulto 

Ito check the acc^aracy aaad repor^cibility of the test 3 some urine 
specimens have been sent to &*o Sievers iA Sto Louiso BTo Sicvers 
performs the teat without knowle%e of the result obtained at the 
. Japanese Hospitolo Vie then eheck his result against oiirSo 

Patient Material ; 

The patient material in this st^ii?.y Is ma<3e xi§ of the private 
patients (JapanesQ mid non- Japanese) of the Japanese pbysicians 
on the Staff of the Japanese EoBpital^ and patients of the Los 
Angeles CJorinty General. Hospital a There ere also aasxe voluatas'y 
non<=patient psrtlcipaaits^ both J&paiese and non«>Je-p»iieseo 

Major Fjadingi s; 

13as essential finding was that the Jepaaese subject? did not 
have a greater freqjiieacy of ©shlorliydria than did the control 
populations o An unexpected findiug was that the 14exie^j ccnt^olB 
diffeared from both the Caucaa-iaaB and Japanese in ha^rixtg excess 
achlorby^ia* !3Sais finding is bssed on a fa^xly small population 
and should be confirmed with rmre datet before acceptance o 

!ISie prepared mss'iiscript (see Part B) ineludes other data and diseuseJoa 
on reprpdueability of the azvsre-i'A tubeless gsustrie analysis test asir?, 
a review of the findings ©f others o 

Sigaifieanee t& Caacer Re search ; 

Developaaent ©f siasple techniques for early detection of gastric 
cancer woiild be an iinporbant contribution to control of this 



23; 



" 



CONKHUiUaOH SHEET - 2 Serial KOo .NCa-157 



iJisease, iJ&tle tutaeless ^latafle sxtalyslQ viil not directly detect 
gastric caneer^ it s^ serve »» a ecresniag loethod to Identify 
persons anKjng ^/hcci the risk of this disease is higbo I-iore elaborate 
diagnostic proce^iircs ean "be carried out wsorxg small higU^riBk 
groups than vould be feasible for the entixe population^ 

Props8e& CoMTse of Projest ; 

Majer effort has been directed toward increasing the number of 
Cai^esi&n and liexiean controls <> It appears so far that the excess 
achlorhydris found msaxyz ^xlcans is perelsting6 



233 



PHS-HIH Serial Hoo HCX«lg7 

Individual Project Report ' ^ 

Calendar Year 1959 



Part B e Honors^ Avarda^ acd Publications <> 

Publications other tbsn abstracts from this project: 

Gr:jispoon^ Lo and Gunn;, Jo E.j, Jr..: A stu^ of the frequency of 
achlort^y^ls- araons Japanese In Los Angeles, JNCI 22;6l7«>531 
(March) 1959 « 



23i4 



PHS"lfXH Serial Ifoo wir^id^h 

Individual Project Report lo Field UveBtigafeionB and 

C lencUtp Year 1959 DeimnetTaticnB Branch 

2o General Field Stuflies Section 
5<. lios Asigeles, California 



Part Ao 



Project Title; Circumelaion and Cervical Cancer 

Principal Investigator; John Eo Dunn^ Jro, Mo So 

Other Investigators; George r^ake, M» Do, Charles Carpenter ^ M, Do tm^ 
lit, PhUip BuelL 

Cooperating Unit: Ibiverslty of Southern California School of Medicine 
DTc Ian Macdonald 

I'^ "iesTB (calendar year 1939): 

ISptal; 1 
ProfeBsional; 2/5 
Other; 1/5 

Project Description ; 

A detailed analysis has Iseen laede of the data on circiisscision of 
male paurtners of cervical cancer patients and controls trcm the 
study conducted by "D^So Edward Jones and Ian Mscdonald during the 
period 1951'=55o Thla aaxalysis shoved no difference in the frequency 
©f circumcision of the male partners of cervical cancer patients 
and controls o This material has been sulaaitted for publication «. 

TOiese findings were at variance with those published by Wycder; 
et alo (AQo Jo Obsto and Gynec, 67:1016- 195^ )i. in which the 
xmcirciaacised male was found to be the principal cause of ths 
excess of cervical cancer in gentile wcaaen^ ^s ccsj^ared to Jewish 
womeno A study carried on by Lilienfeld (Jo Hat^ Cancer Isasto 
21: 715 J, (Octo) 1958) came to our attention, in \rfiich a sxtrprisiiig 
discrepancy was found between the statejaent of men as to circvaacislon 
status and the findings on medical examination o A similar study was 
carried out by Sto Grinspoon, and then by DrSo Drake axA Crpeater in 
Los Angeles with sanewhat slajilar findings ^, Further than this it >rs.s 
found that this discrepancy did not exist for Je^fish sen, circfflaciGlon 
for them being almost always anatomically camplet-e.. On the basis of 
the misclassification men were responsible for in vesssrA to themselves, 
and assusning wcroen would do no better in evaluating their male partners 
it was shown that a case-control ^udy of wGenen mi^t easily fail to 
show a protective effect of circiaacislon even though it exists g 

Part B . included Yes J^ Ho £J 

235 



CCiNllHUAiriOn sheet - l Serial Kod NCS«>l6i+ 



Our conclusion^ then^, is that ttje protection of wcanen frcm 
cervicai eanser afforded liy completely elxci5ncise4 laaies is en 
attractive U^rpotbeeis t&at still Iscks proof, end the usual case^ 
control stiuay In wMeh male circuajcisioa is fetermjne^ from 
querying women on tbcir male partners is unlikely to discover 
such a relet ionsMpo Bie defiDitive stuiSy will involve women 
with cervical cancer wbo have had only one husband and determining 
circumcision status by examination » 

A preliminary attempt to carry on such a stTaoly x/as tried first on 
Los Angeles Co\mty Hospital patients <,, Tisls population was found 
luasuitable because of the frequency of widowhood, broken hcanes^ 
uncooperative husbesnds, etco A ^roup of private patients kindly 
made available by I^, len I-lacdonald will now be tricdo If this 
is xDSxre suceeasful^ other patient sources will be soxight, Pre-=> 
lludnary discussions with some of the physicians of Gynob Clinic 
in San Die^ indicate that they xasy be wllliag to cooperate in 
such a studyo ^eir sizeable accucojlation of cases of e^rlj 
invasive cervical cancer and carclnCTia in situ, as Identified by 
cytology j, would provide a good i^uree of cases o 



Objectives ; 



Detennine whether circvancision status of males is related to the 
occxanrence of ceacer of the ce^rvix ©mong their Bexual partners » 

ttetbods Egg>loyed s 

Interviews by physicians of cervical career patients who had been 
married once only and examination of their husbsjads for circuEicisici 
status o Controls \rill be selected to aiatch on age, racey social 
status^ and ^dietber ever pregnant „ 

Patient Material ; 

1) Private patients of Cr* Ian llacdonald in Los Angeles « 

2) Later^ pcrfjaps, also patients at Gyjsob Clinic in San Di©^„ 
Ilajor Findings ; 

Itone as ycto 

Significance to Caaeer Research ; 

Villi provide an important lead to etiology of cervical cancer as 
well as a BiMple control measure if cirsvoncisioa of the male iis 
found to be protective against the disease o 



236 



COls'TSHUAHOII SHEET - 2 Serial IIOo lICI-l£4 



Proposed Course oS Inject ; 

At present ftrrangeoents have teen made to laterview cervical caacer 
patients and suitable eontrols frcxn the practice of Dr, Purvis 
Ilartin in San Diego » VThen these ecre CQB]gpleted patients frcm other 
physicians in the Gynoh Group will Ise intervieuedo 

Cervical cancor ccrtali^ is only halT as f^quent sbsoiiq Japanese 
vcoan in Calif omia as soong other vcsnen in the state o ArrsngC'^ 
laents arc Txjing nada to exsaine Jsapanese laen to dletennine freqtaency 
of elrcumcisiono Prelininsry evidence indicates that this is 
infSrequent among Japanese meno 



237 



PES-lflH Serial Koo HCI-l£4 . 

In(iivi<iual Project Report 
Ceaeiidar Year 1959 

Part B o Honors J AvJ-ar4s, mid Publlcationo 

Publications otber than etistracts from this project: 

Dvma^ John E„ & Buell Philip: Association of cervical cancer with 
circvimclsion of ©xaal partner.. JMCI 22t7k9''l6k. (jk^rix) 1959 <> 



238 



PES-NIH Serial Ko. MCI-'159 

Individual, Project Report lo Field InveBtigations anS 
CalendBor Yeiar 1959 Demonstrationa Branch 

2p General Field Studies Section 
5o Madison, l/isconsin 



Part Ao 



Project Title; Study of about. 100 cases of polycythemia vera treated 
with P58, 

Principal Investigator a: Paul Calabresi^ Mo Do and Co Meyer, Mo Do 

Other InveBtigator: 

Cooperating Units j University of ifiseonain and the University Hospitals^ 
Depaarfaaeaat of Internal l^tedicine 

lian Years (calendar year 1959)! 

Total: 
Professionals 
Other: 

Project Description ; 

Objectives ; 

' This study is designed to gather BK>re data on this relatively rare 
neoplastic entity« An evaluation of long~term radioactive thei-apy, 
its results and relationship to occurrence of leukeisia is proposedo 
Information concerning occupation^ familial and religious factors^ 
relationship to other disease processes as well as diagnostic and 
, clinical data will be ohtainedo 

tte'feodB Eiaployed ; 

With the cooperation of the Etepartaent of lateraal Medicine at the 
University of Wisconsin and the University Hospitals the patients 
were exaaiined £md the charfcB of the cases under study will t» 
reviewed and analyzedo 

I-lajor Findings : 

Work on this project was completed in 1958 hut three reports were 
published during 1959^ as noted in Part Bo 

Proposed Course of Project ; 

Biis project was ended because of coiapletion« 

i Part B , included Yes ^ Ko £J 

239 



PHS-IIIS Serial Hoo_^X»l?9_. 

Individual Project Beport 
Calendar Year I959 

Part B o Honors^ Awardls, end Publications 

Publications ottoer than abstracts from this project; 

Calelffesis Paul and Meyer, Ovid 0<, : Polycythemia Vera; lo 
Clinical and Laboratory Itanifestationso Annals of Internal 
Medicineo 5O;ll82-1202 (May) 1959o 

Calelreslj Paul and Meyer, Ovid Oo; Polycythemia Vera; XI <. 
Couree and !Rxerapy. Annals of Internal Med. 50:1205-1216 (May) 
1959- 

Sieversj, llauriec Lo and Calabresi, Paul; Gastric Pespin Secretion 
and ABO Blood Groups in Polycytbemia Vera^ Ajbo J* of Dig., ClSo 
(New Series) 1*; 515-521 (July) 1959« 



2h0 



FHS-NIH 

Individual Projecfc Report 

Calenday Yea? 1959 



Lai Ko„ 



NCI-l&O 



1, Field InvestigatioQS and 

DeGsonstratioa8 Branch 

2, Diagnostic Devalopmant Progysm 
3- Providenca, Rboda Island 



Part A,, 



Project Title; Frequency of occurrence of cancer and other aajo? 
diseases aotong a group covered by a state°vide 
uoeiaployiiaent insurance. 

Principal Investigators DTo John B„ Vander 

Other Investigatorss MTo B„ E, Carroll and DTo B. Ko Milmore 

Cooperating Units? Office of Biometry of the NIH 

Unea^loyment Coia^essation ComoiisBipn of a 
New Sngland State. 

Man Years (Calendar Year 1959); 
Total o 3/4 
Professionals 1/4 

Other s l/i 

Project Description: 

Objectives ^ The objective of this project ia to de'cermina tht^ 
prevalence, and if possible to obtain incidence rates on the oecu:c?reii=,; 
of cancer of the major sites among selected occupational groups. 

Methods EB^loyed s In one States unejsployment insusance covers a 
population group ranging roughly froia 236,000 to 300,000c Insuiratics ? 
conwjulsory for all persons who ordinarily cosa under Social Security 
Regulations; that «Duld be most persons %^rking in establisbisientg 
eiEploying four or taore persons, female workers stake up 42% of this 
total, and account for much of the fluctuation in the total sise of 
the group. 

Monthly reports of all prijuary jaalignancies of the luog and 
respiratory tract have been collected by Dr, Vsader sj.nce the month 
of July 1953o Since July 1953 all disabilities occurring in this 
population group have been coded by the "International Classification 
of Disease^" in three digits,, Cases reported to date include social 
security nuisber, age, sex, morbidity code and a brief occupational 
history. 

After review of the problem, it was decided that cancer of all 
sites and major diseases would be collected in this manner. It was 
thought by Dr. Doro that reasonable population figures could be 
estimated froa the census of the State for the groups desired. In 
addition^ on each case of lung cancer ^ an occupational history is 
being obtained by an interviewer froai the Uneajjloyment CoK^ensation 
CoBsnission Office, in conjunction with Ife. Vander. 



2ill 



COKTINUATIOW SHEET - 1 Se«ial No. liCI-160 



Patient Matgri als Working Insured Population of a 
New England State 

Major F indings s None. The study was started late in 1955^ 

Slgnif ic aace to fctie Proggam of the Institute? With very few 
exceptions., no incidence rates £07 cancer or other diseases are 
available on localized occupational groups. Precise infosaation on 
these points ^>ould have definite bearing on further study of the 
etiologies of those cancers believed to be or questionably considered 
to be of environmental origin. Further progress in the field of 
environmental cancer can be obtained only hj the collection of such 
data as described above. 

Proposed Course of the Projects Effective July I5 1957s collection 
of additional asorbidity reports was discontinued. More detailed analysis 
of data already collected is the nest step r< 



Curre nt Statue s An initial brief analysis of data accumulated 
has not suggested any significant trend but a ajor© detailed analysis 
is planned as soon as time permits « 



Part B. included Yes/" / Ko/K 



242 



1 

i 



PHS-NIH Serial IJo. HCI-l6i 

ladividvi&l Project Report lo Fa.eid Investigations mid 

Calendar Year 1959 DscKsnstratlons ESranch 

2o General Field Studies Section 
5o Sto Lottis^ Missouri 



Part A. 



Project Title: Gastric secretory fanctlon in selected population groups 
and eval^iation of tulaeless gastric analysis^ plasma 
pepsinogen determination and Schillings tests as sctreen^ 
ing tests for gastric cancer o 

Principal Investigators : Maurice Sievers, Mo Do; Nell Ic Gallagher, Ma jOq 

end Paul Calatsresl^ Ui Do 

Other Zxnrestigator : Bo Ko Mllmorej, M„ Do 

Cooperating Unit: Veterans Adlministratlon Hospital j, Sto Louis, Missouri 

!48n Years (calendar year 1959): 

Totals 
Professioi^al; 
Other J 

Project Description ; 

Objectives ;' 

1) la selected adult population ^oups^ estimate prevalence of 
gastric atility to produce 

a) Acid 

h) Pepsin (plasma pepsinogen) 

e) Intrinsic factor 

2) Determine the relationship of sufficient eaad insufficient 
gastric secretory functions to each other, to JMiLignant disease 
of the stomach, and to other conditions, ioCo polycythemia vera^ 
pernicious anemia^ and chronic greuaulocytic leukemlao 

5) Deteirainc the Uiiefulness and practicability of tubeless gastric 
analysis, plasma pepsinogen deteimlnatlofis end Schilling tests as 
screening tests to identify population groves \;ith relatively high 
risk of gastric cancer o 

h) Oetenaine -whether the increased incidence of peptic ulceration 
noted by many investigators among patients with polycythemia vera 
and chronic granulocytic letikemia is associated with a change in 
gastric pepsin production or other gastric secretory functions „ 

Part B e included Yes ^ Ho £j 



o li o 



C0UTI1WASI015 SHEET - i Serial IIo, KCI-161 



Methods I^loye A; 

1) Acid-=>produclag ability is 41etermified both by tfee staoSara. 
intubation methoti SBd the tubeless ssatbodj utilizing a,si2re»A 
ion-exchaage resiBo, 

2) ■ Pla^aa pepsinogen is determined by a iEo41ficatioa of Mir sky's 
metfaod (See report on "Indirect Gastric Secretory Studies" by M<, Lo 
Sievers snd George Lo Fischer in Jtoo J. Digest BiSo Kf,S, 2:365*5716,5 
1957). 

5) Gastric intrinsic factor production is estimated by the indirect 
method of 4etern^j3iiig the uptake ond urinary excretion of an orally 
a&Qinistered dose of radioactive vitamin E, (sjodified method of 
Schilling). -^ 

k) Hematologic studies are conducted Including bone marrovf exaialna- 
tionsj (in patients fovaid to have iisdireet evi<feace of a deficit of 
intrinsic factor production) t© detenaine whether peraicioue snemia 
exists » 

5) Gastro«intesttaal x-°ray examination and prolonged follov=.\Tp of 
selected groups of patients are conducted to determioae incidence of 
gastric cancer o 

Major Findings ; 

1) Confirmation of the high degree of aeeuraey of tubeless gastaric 
analysis in comparison with intubation results^ SiS hes also "been uell 
established in all reports in the literature with a coaabined, accuracy 
figure of over 98^^ (l^aSO controlled teBts)o 

2) Aehlorhydria (detected by tubeless gastric analysis and oonfinx^d 
by tube procedure) ha^ been detected in 16^6 percent of all male 
admissions to one hospital laedical ward, ©■^^r a one-year periodo 

5) TKie mean plasusa i>epginogen vali^s have been elevated with active 
duodenal ulcer, diabetes laellitus on depot insulin, or asotemiaj and 
the mean results have been below those of the "normal" gsfoup in 
patients aith gastric carcincseaj, radical subtotal gastrectos^y^ and 
achlorhydria (regardless of other disease (sndi1±>ns)o However >, 3ji 
all of these conditions studied there was sobs overlapping of results^ 
thus limiting the diagnostic valixe of the test in the Ijadividual patient 

k) All patients with acblorl^dria had plasma pepsinogen values either 
belcw or within the normal range ^ unless azotemia was also presents 
It appears that an elevated pepsinogen result in a patient with achlor- 
hydria is diagnostic or renal inaufficiency-"Or of the achlorhydria 
results being erroneous (icCo the patient does not have persistent 
achlorhydria^ althou^ it may occur intermittenly) » 



COHTIRUATION SHEET ^^ 2 Seriel IJoo l^TZ^-lBl 



5) In a large hospital population iVequcncy of defective gastric 
secretion: AcMorhydriaj, 8S/jj 1o\i plasma pepsinogen., 12^; mid low 
Schilling tcets^ C^jo 

6) , Caucasians eund Negroes hs^ a similar frequency of Impeired 
gastric secretory function « 

7) Acblorhydaria emd low plasma pepsiaaogen ocsurred more frequently 
among subjects aitb "blood group "A" them "Cf „ 

8) Plasma pepsinogen concentration vas not significantly different 
for control subjects or those with etoonis granulocytic leukemia or 
jpolycythemia vera^ 

SlgiificBPce to Cancer Besearcb ; 

It ka£ been kno\m for many years that gastric cancer occurs muGli asore 
ftreqwently (about aeven times) wiien achlorlaydria is present, acl^lia 
of pernicious anemia further increases the oceirrreace rate of maiisoancy 
of the stomaeho More acctira'fce surveys of Incidence of achlorhydria 
and achylia and tdie longtime follo^r-up of the icdividuals with ^^tric 
secretory insufficiency ia desirable to develop vcove detailed infoi-ma- 
tion on the relationshipo The simplicity of the iadireet techniques 
(tubeless s'is'&s'ic analysis, plasma pepslncgsn determination assd 
Schilling test) makes them especially desirable for this stu&y„ "Sxis 
project \jrill also evaluate the usefulness of indlreet gastric secretory 
studies as a screening technique for identifying the individuals most 
apt to have or to develop gastric cancer » It will evaluate whether 
•further definitive tests in this aehlorhydrie aad achylic groups are 
a useful combination'»procedure fa* detecting gastric cancer.. 

Patient Material ; 

1) Selected patients witti and without gastric syj^tcats vho are seen 
on the medical Hematological ©ad gsstro-intestinal services of two 
or more hospitals in Sto Louis, Itoa 

2) All patients with a well-es-tabliehed dis^osis of polycythemia 
vera and other hematologic disorders at selected tospxtais associated 
with the University of Wisconsin and Washington University Sebools 

of Iledicinec 

Proposed Course of Project ; 

Uhls project report for 1959 is suJaaitted in order to place on record 
the publication's during 1959 i* of three manuscripts ^ 



245 



PHS»KIH Serial Noo KCI«l6l 

Individixal Project Report 
Calendar Year 1959 



Part B o Honors^ AwardSj, and Publications 

Publications other than abstracts from thts. project: 

SieverSj Maurice Lo^ M<. Do: Hereditary aspects of gastric secretory- 
function: Pace eOidL A£0 blood groups in relationship to acid and 
pepsin production-c Am. J. Itedo 27:2iM5*S55 (Augo) 1959o 

Sievers^ Maurice Lo^ Mo Do and Calabresi^ PsbI; Mo DoI G&stric 
pepsin secretion amd -ABO blood gjroups in polycythemia verao Amo 
Jo Digest, Dlso (Hevr Series) 4:515-521 (July) 1959» 

SieverSj, Maurice Loj Mo Do and GaJJLagher, Neil !»: Interrelationship 
of acid, pepsin, and intrinsic factor prodiaction « ea iadireet gastric 
secretory study o Gastroenterology 57:lc2-l87 (Augo) 1959 » 



246 



PHS-HIH 
Individual Project Report 

Calendar Year 1959 



Serial No, JjCI '^l&J,^ 
Ic Field Investigations aad 
DeaionstrationB B&'auch 

2. Diagnostic Development Vrogxam 

3. Walpole, Massachusetts 



Fart A. 

Project Titles Spectrophotometric analysis of leukeaiic white cells. 

Principal Investigators Dr, John B., Vsnd&T 

Other Investigators? DTo B. Ko Milinore and Dr. R,, L. Smith 

Cooperating Units; None 

Man Years (Calendar Year 1959) s 
Total g 3M 
Professional; 1/4 
Other ; 1/2 

Project Description? 

Objecti ves ; To study and coispare the spectrophotometrie pattara 
of leukemic and normal ^Ite blood cells with the view of using this 
as 3. diagnostic test and a guide to therapy. 

Methods BiBj>lo yed g ¥hite cells are prepared with perchloric acid 
and the extract analyzed in a Beckmaa spectrophotometer with especial 
iate&e£(; la iae &b»0£r^i:ioa peak at 2600 Angstrom unitSo 

Patient Materlal g In and Out Patients of Fondvllle Hospital 

Major Findings 8 White cells frosQ patients with chronic lyspliatic 
leukemia gave approximately twice the optical density readings at 
2600 Angstrom units as cells from patients with chronic myelogenous 
leukemia on the basis of equal volueies o£ cells c 

Cells from patients with acute blastic leukemia gave an even 
higher readiag. 

Signif ieaace to th e Program of the Institute; This proceciurs 
may prove to be of diagnostic in^ortance in distinguishic,^ bativsaa 
lyiaphatic and myelogenous leukemia - something that is not no^» altjays 
possible especially in stem cell types of diseasec 

It may also be of real value as a therapeutic guide possibly 
giving an early clue to impending relapse of disease. 



2^7 



GOOTINUAXION SHEET - 1 Serial HOo....... J£l;-i.; - 

Propos ed Course of the Pro.| ect:; Progress is siizsaarised in a 
recent publication (see under Part B) which was published ia Cancer 
Since the date of publication fiirther work hzs been done ueiag a 
method of dialysis in an attenij^t to aiot-e sharply bring out Che 
differences between esctracts of cells of lya^hatic end oiyelogenous 
leukemAc It is too early to draw any conclusions from this lattei? 



Part Bo included Yes/X°T 



248 



PHS-KIH. Seri: 

Individual Project Eepo?t 
Caleradas Yea? 1959 



P ar fc_ B_ . Honors g Awards, and Publications 

Publications other than abstracts £rosi this projects 



Vaader, John B„s Optical density messureiaents of leukemic cellSc 
Published in Cancev , Marcb-^ril, 1959 



Honors and kvexAe relating to this projects 



^'-(J 



Individual Project Report 
Calendar Year 1959 



lo Field lavesttgations and 
DeaKjnstrations Branch 
2„ Diagnostic DevelopEsnt Programs 
3<, Walpole, Massachusetts 



Part A. 



Project Titles Electrocardiographic Chaages Associated with 2 Million 
Volt X-Ray Tfeer^y to tha Chest. 

Principal Investigators? John B. Vaader, Mo Do 

Other Investigators; 

Cooperating Unitss Pondville Hospital 

Mao Years (Calendar Year 1959); 
Total s 1/4 
Professionals 1/4 
Other s 

Project Dsscriptioas 

Object ig'ge s (1) To determine the type of electrocacdiographtc 
changes which eccvr ufeila patients are uadargoing high voltage s^ray 
therapy to the chest for malignant disease, and their frequency „ 

(2) To attes^t to relate these changes to etiology with especial 
reference to cardiac changes due to K-^ray therapy itself « Special 
consideration will be given to EKG changes due to pulnjoaary easbolus, 
myocardial infarctioe, or positional changes of the heart due tc 

such factors as decrease in size of adjacent lyiaph nodes. 

(3) A preliminary study showed that the changes in electrocardio- 
graph tracings are fairly frequent during high voltage s-^ray therapyc 
Evaluation of the etiology of these changes would be of isisportance 

in providing a therapeutic guide. 

Methodis Saaployeds (1) All patients w5.11 ha^'-e at least two ba':: ;- 
line ESG's before treatment » 

(2) Daring treatment they will have daily EKG's and after 
treatment EKG's every 3 days until the tracings are stabilized. 

(3) Before treatment all patients will have careful evaluation 
of their cardiac status to rule out pre--eslsting heart disease^ 

(4) All patients will have x-rays of the chest bafore treatment 
and at appropriate intervals during treatment. 

(5) Additional laboratory studies, such as transaaainase deter-- 
miuations will be made as indicated,. 



251 



GOOTHTOATIOK SHEET -- 1 Serial No. ?^2^': 

Pstient BJaterlal ; In-patieats of For.dville Hospit&l 

MaJ og_giadlQgs? Serial alectJfocajrdiograajs have now been takea 
on approsimately 20 patients receiving 2 mil lion volt x^ray treataent 
to the chest. A significant atjasber of these showed definite chaBges 
during treatraeist. but a final analysis of these has not yet beea iBsde„ 

jSigaifieaace 'cc the^ Frp s^^^ Q^ ^^^ ^ ^sfc^^.M?g. ° Since chest i^-aia 
and tachycardia are freqsiently considered indicatioas for stoppiag 
radiation ther^y, more knowledge about the EKG findiags ia thsse 
patients might form the basis for sore accurately judging asiount of 
treattoent and dose. 

PFoposed Cpursa of Projects This study has now reached the poiat 
where the data so far aecusaulated sbus£ be analyzed. My present iss" 
pression is that there is sufficient material available to give con- 
sideration to the preparation of a manuscript for publication. After 
a preliminary review of this nsaterialj however , we may decide to 
continue the work until a number of additional patients have been 
studied o 



Part B. included Yes r°7 ' Mo/FT' 

251 



3(^x31 Ho„ ^„^__ W^Jx: A^2..^ 

PHS-MIH I„ Field Investigations and 

Individaal Project Report DeaxonstratioES Branch 

Calendar Year 1959 2. Diagnostic Developmeni: Progrei 

3, Waioole, Massachusetts 



Ps'oject Titles The relationehip between casfcinoma of the endomet^iusi 
and diabetes mellitiiSo 

Principal lavestigatorg Dr, Jolm B, Vander 

Othe? Investigators^ Hone 

Coopesrating Ualts; Foad^ille Hospital, ^alpoleo Massacbasetts 

Man Years (Calendar Yea? 1959)? 
Total t 1/4 
Professionals 1/4 
Otheff s 

Project Descriptions 

0bj.ectlve6g To investigate the allegedly higher pi-avaleace of 
diabetes saellitus aroorg patients with carcinoma of the endosaetriusJc 

Methods Easp lo^d s Records of patients with carcinoma of the 
endo@etrii2Si at the Foadirille Hospital in Walpole, Masso, wese reviewed 
to determine whethe?? such records also indicate the presence of dlabefceSc 
The diabetes pretraleace rates of these patients compared with diabetes 
prevalence rates reported by surveys of the general population In 
Massachusetts.. 

Patient Mategials In and Out - Patients of Pondville Hospital 

Ma.jor_Fiadings s There was no significant difference between 
diabetes prevalence in the study group as conspared with a control 
population of similar age, sex^ and raceo 

Siffliflcance to ^the Projij^gasi of ^the lastitute; If there was a 
relationship between neoplastic disease and disordeffs of cc.rhoh.jd.~cta 
metabolism, the desnonstrstion of this aseociation would help to clarify 
the pathogenesis of cancer. 

Proposed Course of the Project s Project has been cosspleted. A 
manuscript jreposrting the findings was published by the ggser 1 c an ^^ Jou r a a I 
of Obstetrics and Gynecology , February, 1959« 

Part B. included Yes/F7 Ho/"°T 



252 



PHS-SIE Serial Ho. _J^z}^. 

individual Project Report 
Calendar Yeas- 1959 



Part B_. Honors, Awards, and Publications 

Publications other than abstracts from this project; 

yarsder» John B.s The Associ^itica of Siabetes I-Iellitus and 
Carcinossa of the Eodometriusao Published by the Aas eglg&n 
Jougaal of Obst etgjj:jLagL4--iypecol.og;y , February 19 59. 



Honors and Awards relating to this projects 



253 



PHS-WIH 

Individual Project Report 

Calendar Year 1959 



Serial No, __„_NCI^16J_ __^ 

ic Field Investigaelous and 
DeEsoastrations Bs'anch 

2, Diagnostic Developjcetti: Prog?ram 

3, Walpole, Massachuaatta 



Part A„ 



Project Titles Amino Acid Determinations of White Blood Cells, 
Using Paper ChroBsatography 

Principal Investigator? John B. Vanderj M«Da 

Other Investigator I Mr, Karl HesSj COSTEP 

Cooperating Units; Fondville Hospital 

Man Years (Calendar Year 1959); 
Total ; 3/4 
Professional: 1/4 
Other ; i/2 

Project Description: 

Objectives; (1) Ttie insmediate sisi is to daterraiee whether 
there are differences between amiao acid patterns of leukemic cells avc 
those of no^fflsl white cells %^ich can be utilized as a diagnostic aid 
in establishing the presence of leukemiso A further aim is to utilise 
this method in sn attenspt to establish tlta exact type of leukemia 
with a special reference to the sfcaia call leukemiss which ordinarily 
cannot be satisfactorily differentiated by the usual histologic siathods 

(2) A more basic objective is the determination of assino acid 
patterns with the ultimate aim of labelisig certain amino acids wlth 
radioactive substances so that these metabolic materials can be folio.:" 
It is hoped that such information might be of value in suggesfciug nGc.- 
types of chemotherapeutic agents » 

Methods Biaj?loyedg Amino acids are sKtr acted from white cei'.£ 
acetone as a solvents Fat is reasoved with ether, and the. rasulfcing 
easily eatractable astino acids are concentrated by exj-aporation o£ the 
solvent. Amino acids are then separated by means of paper chroaiato-: 
graphyi at the present time phenol and butanol are being used for twc- 
dimensional separation. 



Patient Materials Patients at the Pondville Hospital in 
Walpole, Mass. 

Major Findings; Wa are continuing to make determinations ox 
amino acid patterns on normal and leukemic cells and have nox-? conipleted 
about 30 of these. Although some difference in amino acid pattern is 
noticeable the findings are not yet sufficiently conclusive for an 
opinion of the value of this procedure. 



254 



CCKTIKUATIOi:^ SHEET - I Serial Ko„ jSCI-167 



Significance to the Proggam of the lastitutas As stated under 
Objecti'f/eSs aay additional method which would allow different iatioQ 
between normal and leukemic ceiis would be vesy useful especially in 
such conditions as myeloid metaplasia and leukessoid ^eactionSo Of 
even greater value would be a £'eliable isethod %Fhich ^wuld allow exact 
classification of the stem cell leukeaaiaso 

Pgopose d^gugs g of t he Pgoject ; It is propored to continue sssinc 
scid determinations oa normal and leukemic cells end also sera from 
the sajue patientSo 



Part Bo included Yes/ / Ko/^^T 



255 



AKHUAL REPCST 
CAIfiKDAR lEAH 1959 

Serial Hoo NCI °1< 
Qraats Section 
Field IiweBtigatloaas and 
Denoostratlons Branch 
Rational Cancer Institute 

Tbs Grants Section Is respoi^lble for tbs administration of the 
folloiring programs: 

(1) Field Investigation Grants 

(2) UadergradxiatQ Cancer Training Grants 

(3) Cancer Teaching Projects 

ik) Cytology Technician Tralneeships 

(5) Clinical Traisae®shlps 

(6) Sadiuia Loan Progreffii 

(1) Field iB^^sti^tiegi Gz-aats Rrogram 

Durtag tho calendar year 1959 there vere 18? grant applications, 
totaling $fe, 241,039, coBSidered by the Gaacer Coatrol Cfflaaaittes uod ths 
Katlonal kdyi&ory Caac@r ecsaacll. Of this nuaber 90 grants totaling 
^>265»738 wsr« saw and 97 grants totaling $1,975*301 wsre continuation of 
old gyaats . 

"Hie allocation of the funds for the Field iBvQBtigatlon Qraats in 
1959 vere apportioned in the folloving categories: Cytology grants ajsd 
Xirmstig&tiox^ and BeiBonstrations graats other than ecology grants. 

Of the 187 applications siibmltted, lltO grants totaling ^,6k'J,k80 
vsse approved - kf Qytology grants for $96^,1*23 and 93 other graats for 
$1,633^057* Of this nui^er, ^3 grants totaling $690,355 vere nev studies > 
11 C^^tolo^ grants la the amount of $l6o,^^ and 32 other grants in the 
amount of $^9,713; 9^ grants totaling $1,900,357 wsre continuation of 
studies already undffiffway - 35 Qjrtology grants totaling $785, S^H and 59 
other grants for $l,llfe-,5l6j 3 grants totaling $56,768 war® deferred grants - 
1 Cytology grant for $17,9^0 and 2 other grants in the ioount of $38,828. 

Forty-one (1*1) graata totaling $1,375,210 iwre disapproved - 
26 Cytology grants in th© aiaount of $778,815 and I5 other grants in the 
amount of $596,395. Six grants totaling $218,3^9 vere deferred - 2 ^rtolo^ 
in the Jiasoxmt of $67,326 aad k otJ^r graata in the aaouat of $151,023. 

Tbsr© ^©re I5I Pi^ld Investigation Grants totaling $2,889,021 vhich 
's^gre active in 1959. Of this aua&er i^O grants totaling $801,282 were 
Clinical Investigatioi^ in the folloviHg fields of jfose jpch: 

256 



- 2 - 

22 Clinical lisvestlgatione la general $380,676 

16 Radiology i|00,391 

1 Sisrs^iy 6,996 

1 Eserapy 13,219 

Forty (to) graats totaling $960,8^6 vere in the field of Patholoffir 
in the followlag categories: 

9 Pathology in gas»ral $32^,710 

25 Cytologr 669,950 

6 Magjacsstic l66,l86 

Twenty-alBS (29) gz-anta totaling $fe50,lt83 w^re in tbe Education 
field in the following categories: 

20 Iducatioa in general $305, 361 

9 Cytology training lk^,122 

OMrteen (13) graats totaling $183,703 were statistical studies; 
6 grants totaling $136,9^ «er« Snriroosiental studies; 22 grants totaling 
$3^,795 "^3^9 Bpidsfisiological studies ind a suj^liEental grant of $15,000 
vas addM to th& Chalnsian's grant to allov for tbo p^mcer G ;ntrol Ca2&aittee"8 
ea^ffiasea diiring 1959. 

Six. (6) project sits visits vera ssade by neosbers of the Cancer 
Ccatrol Co^d.ttee aad staff representatives to 6 locations. The pxnrpoee of 
the visits va@ to c^tain additional infestation for the svaluatioo. of aev 
applicaticma, to detesmine adsquacy of research facilities, ccaapetency of 
applicants and professional staff, and to determine progress aM sigolficance 
of studies req,ulrln3 ctmtioued or additional financial support. 

A total of 25 studies or investigations terminated during tfas year. 
Eowsver, to date, only five reports on the final results of these stiidies 
have been received. 

The final report on the @rant for the supp<»^ of tbs publication 
of a brochure "Exfoliative Cancer Cytology for Practicing aysicions and 
Medical Students" vas received fSrcaa Dr. William 0* Russell, Chairman of the 
Zsi?o(£i£^tion Coaasittee, College of American Pathologists, and Cbaixman, 
Xntersoclety @ossaittee for Xofonaation on Pathology. 

This publication 'Kas writteia and distributed to ail active 
physicians and senior nodical students in ths IMited States. Tbs brochiire 
vas an eight «pa^ publication, vas profuseily illustrated, and vas printed 
in full color. It vas tiritten under the direction of the Coesaeittee on 
Exfoliative C^tolo^ and the Publications Coe^dLttee of the College of 
American Patholo^sts. Tbsse cosaeittees fsere responsible for the editorial 
content, the ill\istratiQn asA. the foonaat. 

Approsciisately two hundred fifty thousand copies of ti^ brochure 
were jointed. IMs presided copies for all active physicians and senior 
ssdical students in ths U^ted States; and furtbar, provided for aaailing 

257 



- 3 - 

copies to all mnr senior medical students each year for the next five 
years. In addition, a few thouaiind copies are available for mi«cellaneou« 
mailings by the office of th® College of American Pathologists. 

Dr. Douglas P. Murpi^, Etaeritxis Profeaaor of Obstetric® and 
C^jmecology, School of Medicine, University of Peaanoylvania, submitted his 
final report aa a fivs-year genetic study of "dancer in Families, A Study 
of the Relatives of 200 Breast Cancer PrcibandB". I*e study was desi^ied 
to obtain inforaaation oa two questioas: Does the disease occur more 
frequently in relatives of a person who has cancer -'•Jsan in relatives of 
a person t^%o dees not? If there is a difference in the fr«quiency of 
occurrence in these two ©coups of relative*, is th® difference greater 
in soffits ralationabips than in others? 

The study pcpulatioa was coBgpoeed of two groups of families, 
one in ^aich a living prdbaad had cancer of th« breast, and the other in 
•v^iich a living pro5>aiid did not have cancer. The study dealt with two 
geioerations, that of tlse proband and t^t of the parents of the prdband. 
The cancer probands vere ^Aite vosaen between ages kO and 65. The control© 
vare living vhite vosasn vho vera beti^een agses kO and 6$ at the tizas -Uiey 
registered aa dent^ clinic patients. 

The authors vero of the opinion that tbe control aad study 
populatioeas are not coRparable to the degree that can he achieved in 
either a prospective or a laboratory invest igat ion, but thiy believw 
that tbs control populatiora resea&les the caacer population @o clodtely 
that ajay differences %^ch aslg^t be fouad between tham woiad b® real 
differsinces, aatd not tb9 r«sult of atsy bias In either the selection of 
the controls or the collection of tbs infonaatloa. 

A coEseparisca of the freq[U£i;icies of cancer in the ca&cer and 
control relAtivss leads to ths follo^ring coxtclueioos: 

(1) If a familial teadMicy to develoip cancer did exist, 

it yias not large enou^ to be detected in a study of this size, 

(2) In those iniTtaiiccs in ^bich there "miB a statistically 
si^iificaixt differenc* b«t«een the cuacer and control groins 
vXth r««pect to th® frequency of cazicer, it va« found in the 
TBsarm r«aote rat£»r then in the closer relatloashipa to the 
prd>aad, and the hig^r froquancies of the disease appeared 

in the control rs-thesr than in the cancer groups. Such findings 
do not constitute evidence of any familial teBd«Bci«A. The 
best explanatioa is that thm^vere occasional chance differ- 
ences «hich the sii^ficance test criteria label j^e statisti-' 
eally significant o 

DTo 3rian MacMahoa, Ospartasnt of Eavlroomental Medicice and 
CcBsaunity B©al.th, Stat® Itoiversity of Hew ¥ork, eubaitted his final report 



258 



on the follo^Qfi studies: "Studies of the Splcbeslology of l&a!s!sml& asd 
Other Reoplaetic Dieorders" aM "0^ Lactatloa Histories of ^tients with 
Breast Cascer". DTo Mat^ahoei has made the followlsig nxxesmry statemsnt: 

X-rsy ssd otJ^r asedical Mstorle® have been d>taiaed for 129 
leuSESsaia patients, sasd ctssgjarlsons bave bess aads of thg histories of 67 
jsatchsd patisnt-relfitlve pair® and of 7^ sa&tched leujcomla patient- 
other patiesst pairs. So aoteworthy differgnc«s have been founds but ths 
numbers are di©app6iatiagly acaall, 

A cohort stiidy of the relationship bet^seen preoatal X-reys and 
chilciliood cancer has been c<2sgpleted as plasjaed, with no difference noted 
in cancer iacideace bet^a!t»n children irradiated in utero and those not 
irradiated. This stti^ is belsg expanded substantially under contract 
vlth the Ato^c Isssrgy Cocealssion. 

%52 patients with breast cancer have b®en intervieved and ccsa- 
parisons loacte bet^sen 3^ of thsse patieiats and ^bO laatched general surgical 
patients. Ho dlfffersnce in nursing (lactatloa) histosy w®re ©vldeHt, but 
the breast caacer mtl®sts ^re si^dficasstly fe^er histories of artificial 
B29norp4&xia@ th^ did t^ gsaaere^ surgical patients ;, lutplylng that artificial 
Eseoopause vs&j protect to soaut extent e^inst the de^elopflBfrnt of breast cancer. 

A sodiflcatloffl of the imxisjun likelihood isethod applied to log- 
noxnsl sus'Ti'TEd. &&t& has been developed and applied to data on chronic 
leuJsesda and sultlple igyelaEee. 

A stiidy of t!m ethnic distributioa of cancer aortality in S®*f 
?0(rk City in 195^ h&s been cos^leted aad date, tabulated on distribution 
by Eealth Area of residence and occupatitm. 

Ibe final report has a list of 12 publications vtalch were aided 
by this grant. 

A tersilnaX aiad susaiary report on ths cytologic exasilaatloas 
of sputuia as a screenlss procedure for carclnoaa of the lung ^^sls submitted 
by Dsr. 0. D. S^th, lKal'9@nslty of Kansa^s ^iedlc^JL Cezttor. The Initial 
isvedttigation vas begim by &r. Ann Pollak sM has received three years of 
^raat sirpport. 

The objective of th^ study vas to devise methods of dbtaiciog 
sputa for large scale screening and stiidies iztllialng an exsufflator or 
"cough Bachin)s"o I& ccs^>arison vltb normal cou^blag it vaa found that 
the exsufflator or nonml coughlilg was la no «ay reli^le or predictable. 
The plans to search for a reasonably reliable collection eethod such as 
inhalation of a single posrtion of ether raporg to caiise a forceful cou|^ 
to obtaia a aore reliable sputua ^i^re.not carried out. 



259 



» 5 " 

|2) Utedsr^'adiiatc Ob slcs t TsisJislsg, Grant Froggam 

This year (1959) is the twalfth year of qperati<» of thiis program, 

IXsrlssg tMs psriod therm war® 137 Undergrafiuat© Caacar Training 
Grant applieatioaa rsceived, totalling ^,M4,438 which waara approved by 
the Caacer Ccatrol Cc^^itte© aisd tix» Hatloaal Advigory Gaacor Council. Of 
ciB^sr 85 «sr@ to sssdieal schoola >&ich included 82 k-yamc BMical acfeools 
oligibl© to receive a BsaxlKusa of $25,000 aanually, and 3 2-3?ear aedical 
schools eligible to r®c@iv® a laaKiiEUHi of $5,000= Tbs total aaaount of 
su^ort to ^@dical schools ^^ ^f06k,k6&. 

'ThBrm «®re k£ d®atal schools eligible to receive a iwaytn^ffli of 
$5,(X)0 aaaiially, totalis $229,950. 

Th©re ^ses® 6 osteopathic sctaools eligible to receive a maxiittvaa 
of 125,000 aajsu&lly, totaling $150,000. 

Tbssr® i^TSi ait® visits ra^de to 11 of tha assdical aod dental 
school® diiriag 1959 by. i^sibars of the Cajaeer Control Comaittea and the 
National Cancer Izmtitut® staff, ^l^siesa schools have not been rieited 
since 1953- Tbe«e visit® w^e to evaluate t^ efffectiveneea of tbe pro- 
Sswm. of a school and give the Dsan and teacMag staff an qpportvuaity to 
esqpress an opinion on th@ xx^& tor continuing it, on the value it had b^®n 
to thaa, esnd hc^ it ^iould be vised if contiimsd, 

A sursBy '^is s^d® of tb& lasdical, dental aM osteopathic schools 
by letter, asMi3^ the ISsans for en assesss^nt of i^ contribution '«mch 
th« ^ant of $5,000 a®a $25,000, s««5«ctively, over the past years has 
Qad» and ajsking th@si if they ^fould ccraaeat as to the de3i3rabill1^ in 
their school of increasing the amount of the grazrk as follows: 

(1) M«-year s^dical and ostecopathlc schools free $25,000 ixp 
to $35>000 

(2) 2«y®ar osedical schools frcaa $5,000 vap to $15,000 

(3) Ssst&l schools froa $5,000 to $10,000 

Tb® results of the survey vers reported to the Caacer Council 
in March. The replies to the letter on the pressed Increase vere as 
follows: 

80 of the £S fouToyear saedieal schools vave in favor of th® 
increase. In addition, four of these schools had the following 
i&aaxka on the Increase: 

(l) 0^: school Stated that the level should be Increased 
. to $50,000 



26Q 






. 6 - 

(2) C&^ school stated that azx additional $^,000 shoiild 
\>9 glvesa to the larger schoola 

(3) Oas school would ha in favor of th® iacraaa® onJy oa 
a cotapetitlve basis 

(k) OsQ£ school vould be in faTor of ths increa«ffi if per- 
mission is graatsd for ^ipemid to stiideats for partici- 
pating In or beccsBlatg ac^xmlnted vlth research. 

All 3 of tbs tifo-ysar nodical schools ware la favor of tb» Incraaoe. 

Ii2 of th& k^ deotal schools -were la favor of the increase and 7 
of th@«e schools felt the level ehould be increased to $l$,OdO inat«ad 

of $10,000 a 

All 6 of the osteopathic » hools i^re in favor of tt» increase. 

A eursaary of t^je relies from th® 129 of the 136 institutions 
questicxisd on the proposed use qf tha additional fuxkis vere in the follotfing 
categories: 









Establish BTo 






Redesign & 


Sfev Phases Report 


Xacreaee in 


iBcrwase la Additional 


Strengthen 


of on 


Persojaael 


Salaries Bcp;ilp>sient 


Program 


Srogras Sesilnars Use 



76 15 32 37 23 15 10 

The reqixsets for increasing this grant to each school had been 
under study by the Caacer Control Ooranittee and a special ad hoc ccsnsilttett 
of thfit £^oup for tt^ pest tvo years. The Coasalttee recoiraended to the 
national Advigosy Cancer Council an increase of $10,000 for each ^-year 
oedical and osteopathic, and each 2-year ssdleal school, and a $3,000 
Increase for the dental schools. "Rm Katlonal Advisory Cancer Council 
approved the increase, as reccsasssnded by the Coesalttee. Follcwisg this 
recoiasendatlon, the isscrease vas requested in the budget for 1961. 

(3) CMacer Tea.chiag Rrojects 

Baring this year there were k teaching projects totaling $81,396 
supported from Training Grant Funds. Two (2) of these were to Schools of 
Public Health, and 2 to !!ursing Schools. 

The @rent to the Ifialvsrsity of North Carolina School of Public 
Health in tt» anouat of $1%,472 i<as designed to strengthen cancer teaching 
in the Public Ssalth currlculxsa, particularly in their chronic disease 
teaching program. Hot casly will the students studying for their MoP.Ho 
degpree receive the benefits of this added ^i^gthasls, but Public Health 
workers throu^ the area served by the school will benefit. The gscmit to 
the Johas Hopkins School of Public Health in ths asount of $35,000 with a 

261 



- 7 - 

ccssmltJBSnt for fiv@ yesra Is iised to emphasize clinical dieeaee tsacMistg 
aad to providQ epaclal p<^tsraMiTiste tralalag facilltlsB in cancer asad 
coss@ctlv® tl®su@ diseases. In tbe^e respaeta the grant is dssigtued to 
broadaa asd atoplify th@ tralnias program in chronic dlS9as«@ nov in 
ofperstioa at the Jcftana HqpkLas School of Ptablic Health. Two categories 
of i^sicismis trf.ll ba in traijslag: (l) Pulblic Ssalth Officers yiho ar@ 
csadidatde for M«P.H.; ai^ (2) full tlise felloes in chronic diseases. 

'Bm mar slag grant to Ksu York Uhivereity in ths amount of $12,9^*3 
is to devslqp professional nurse Isads^s in tha area of cancer nursing 
throu^ pro^-aais of specialisation leading to JfeuBter's and Doctoral Degrees. 
Tb® EBjrsiag grant to IMi'^BX^ity of California is th© amotint of $18,983 iB 
to help establish a xssv cancer training program. !]^is pro^r^a is to be 
directed to tte follosjlsjg: 

Group X Hurees caxlng for cancer patients and/or exipervlsing 

others 

Grot^> I! MuTBes teachixsg in nursing schools 

^txup 1X1 ^urs@s ia graduate-degree prograios 

Workshops azjd institutes of oiaa of three weefes duration for the first two 
groups are plsjxosd, and for the third groisp, votrlEing toward a degree ; the 
training vill be was® intensive, and vlll include studies euch a« social 
sci encee and stat isti cs <. 

Th© Cancer Testing Program ©imported for ths past U yeara 
throu^ grant CT»5077(C3jO) was discontinued by the Naticmal Adviaory 
Cancer Couiicil. Ibis action vas t&ksa on the grcmnds that the objective 
of the study had been realized and further support vould be of little 
additional value. 

(k) Cfytolo^ TecbaiciaB Tt-ainaeahipa 

!EM9 program \ms activated tvo years ago to meet the seed for 
technicians in exfoliati-ye cytolo^ for research activities and service 
prograiaa <. It va» possible to greatly expand this progrem last year vlth 
the ailotsBsent of $l40;000 vMch is the swm for 1959'. 

Tba eatire 19^9 allotisent has beesa coosmitted. This vill provide 
stipend® for 95 positicms which have been aiiarded to 19 training centers 
in 16 states. !£Mrty technicians have been in training during the past 
six laonths, 6^ ha-^e been coEssdltted for ti^ nexb six mmths. Ml of the 
cezsters have six manths training coxxrses in uterine exfoliative cytolo^ 
except o&e ^ilch giv^s a nine months course of training for all body 
sites. Site visits v&se ns&de to three of tbese centers during the yearo 



262 



» 8 - 

(5) Cllaiegd. Tyaii^esfalps 

TMs year isarka ti» ZZad year of oper&tioa of this program. 

Duriaag the caleiiaar year tbare were 231 graduat« pl^slclaaa 
traiasd ia ^ participatlffig institutions at a cost of $9^3,765, uM©r 
tMs program fcMP ^riods Taryiag frca four Boaths to a fall y»ar. Kieae 
jdJO'Sieiaiis WS2?® t?aiJ3©d la th« follo^i^ q;»eclalti9e: 

Siffgeiy 71 

Radioloejr 70 

Pathology 53 

Cbstetrice & 

GunascolQgjr l4 

Ifitemal Mesdlcine 23 

Of this auBiber I9I pbysiciasos are eiurreaitly in trainixtg, 3it« 

visits ^ss's^ msdis to 5 of ths l&stitutlosus uhere th9$« ph^iclana are 
traiaijiSo 

1%sr® \m,» aa. iijcreasa In this year's appropriatioa oTor 19^9 
in the amount of $2^,000. Tim isscrsasa vaa not approTad for use imtil 
Octciber 16, 1959= Since mor® than oaa-fourth of th© training y«ar for 
realdents bed passed before th^se fux^s wsre availebls^. siaoy of tha 
applic^its eithar had received support fs-oia other sources or decided not 
to continue training becaus® funds could not be found to begin or contixsue 
training. 

Thar® has be«n an increased d^aand for training positions. 
ApproxiBiatsly 50 nev requests haTe beea received^ laost of vhich ar« frcoi 
Isfititutlons that have not previovisly paT-tlcipated in the program » Mo^t 
of th)S institutions that have eponsotr^ and trained residents have 
requested additios^al positions. This has occurred even though thsy have 
been ad7ls@d no increse® in fwadiB is expected. 

Plane ere eov under vay to convert the clinical traineeship 
program f?oa on® of direct su^rsport and individual residents to a grant 
type pro^^Ei for cancer training in departeeats of hospitals and institu- 
tioae having s^cogsilzad or approved training prograias. litalniag will be 
in -yje saaia specialties of raediclse as iu the pa@t« An ad hoc coioaittee 
is^ittlzig is schedule for Jaavavy 21 and 22^ 1^0, to formulate regulations 
and procediires fos* opesrating the netr program. It ie hoped that the ehangie 
caa be effected in time to vs3£s great a^iard^ before the end of this fiscal 
year» 



263 



(6) Hadims Loaa Program 

1ti& R^uiitmi Loaa I^ogs^js has qper&teid at tbe saB» lev»l as In the 
past. Fifty iiiistltutions ciirreatly have radium on loan. 

Bm'lisg ^xls yeasr a qu@®tiamiairs v&a 0®iit to sach imrtitutlon 
to Wrif^ the amouat aM "^grpe of radium thi^ had r«ceiv«d and to detsnai^s 
the condition of each unit. The aoalysis of th® questicsumires indicated 
that ns^^ of the vmits %«r@ in poor oar umisabl® conditica, jaasor wsre leaking 
and ecme i^s7# not being i^sed because of dbgK>lat« size. l%sai results of the 
sunn^ 'giex'® fursi^^ed to a nraiber of outstanding Hadlologlsts aad H^sicista 
for ecsQs®nt smd rsccsaasadstioss oa the desirability of continuiag th® 
progg'esa and ths chan^® th&t should b@ Eiade to saodemize ths program and 
iasufg sajpa units far coEtinuous u^ if it wsre decided to keep the progrsia 
in operation. 

"Sc^ cceasultants adirised that all radius^ now in us® ehotild b® 
called in b@cau^ of its unreliability a® a therapeutic absent ^ its obsolete 
s£2«»d ^ckages, b9S>^ isnits had b^en reported aa leaking and neaoy more had 
not been ch©eb?d either for leakage or distribution of aradiuia within the 
uniti 

l&s consultasarbs felt that there ^las a definite need for the 
prograaa to coatinu©. Ho^rsrer, they re'coBEassaded that it b@ done only if 
all radiisa %?as called in aM coav@3rted cr exchanged into atodem type 
packages that could be us®d c«atinuously without fear of leakage or distri- 
bution of radium in each unit. 

A cQgxtract to osstf out these recoassmidations has been requseted, 
Tb&.b contract calls for conversion or exchange of all radium over a period 
of tvo years. It is propossd to have <%aly fi've standard sizes available 
for distribution, as foUovs: 



Ho. of unit9 


Size 


y^ 


280 


Tubes 


573 


10 H@ai 


Tubes 


150 


1 M@> 


ISesdles 


150 


2 Mga 


Seedlee 


52 


3 Mega 


Needles 



All units will ha^re a doiible seal so th&t ths process of application or 
ioplaatatica will not produc® leekage . Sach \inlt vlll be dry-packed to 
prevent c!jaa@® of distribution of radlua. 

Institutions «Ji0 «l8h units of difffesent sizes or types than 
those a^aatloEffld vill not b® pes»itt®d to receive radium. It itf the 



264 



» 10 



opiBicm of the ccasulteats that xmLts of ttess sizes and typ®s will pro- 
vide sufficient flexibility to treat alaoet a^ type of patient %«je2^ 
radixEa is indicated. Tbera are no plaa® to ^^sad the program. 



265 



serial IfOo I»32°l8a 

lo Fl^M^XavestigatiooE abd 

Iffidividval Project Beparb 2p Nu^siog Seetien 

Cal<§zKlax fear 1959 3» Silvey Springy Maryland 



Bart A, 

Pttsj®ct Title: Caacsr l^araing Bibliography 
Principal Investisat^sr; Patricia Bo G«iser 

(Dt5i«r Xav@0tiG?it©r« ; Kons 

C®«^eratins Usits; None 

Maa Years (Cal«sdar Year 1959) J 
Total: 7/12 

Profeseisa^ij Vl2 
Ofehes-; 3/12 

P^>J®et Bsscriptiea; 

Anm>tat@d bibliography on appnsxisiat^ly $00 articles ^ 
bulletins^ b<ook3, filss aad oth@r vistsal aidd azid other 

prostheses o 

Paroposed Course of th® Pg^jsgt; 

It is aaticipated that a et^pleassnt ulU be prepared 
each year to i&clMe the z^v isaterlal which becaaes available c 



Part B iacluded Yes ^ No ^ 

266 



Fart Bo HsfSXf^Sg Awards y and Fubli^.tloas 

Publieatioas "Bifelis^^aphy ©a Censeer f©y Ifas's@s" 

Uo So D@pa^tE^nt of E@alth^ MtSi^tion; and W^lfas^ 

Publie Health Sear*rie« 

Nati©Qal ^28titutes of Health 

Oct©bsr 1959 (PHS Publimtlon U®, 687} 



267 



serial NOo I^2°ia3 
lo FleM iQvestigatiosas axsd 
PES^NIH DsmoQstTatlozut Bra&eh 

Individual Project Report 2e Kiorsing Section 

Calendar Year I959 3o Silver Spring, Itoxyland 



Bsuft Ao 



Project Title: Preparation of a Monograph on Radiation (For 
Ntirses ) » 

Princil^l Investigators Elizabeth Walker 

Other Invest ig&tsrs: Rosalie Xo Petersen 

Cooperating IMlts: Advisery Conaaittee cssapriaed of 
Representation frcan: 

lo Uo Sc Azaay 

2.0 Uo Sa Havy 

3o Veterans Administrative 

4o Mo Do Anderson Hospital 

^o Brookhaven National Laboratory 

6, Cak Ridge National Laboratory 

To University of Pemasylv^nla 

do BosveU Park l^a^rial Hospital 

Man Years (Calendar Year 1959): 
Total: 1/3 
Professional ; 3/l2 
Other: l/l2 

B?oJe<rfc Bescriptiissi: 

Oajectiyes; 

lo TO stake available, in one reference, the 
infonnation vhich a nurse needs to function effectively 
in providing cajpe to patients receiving X"»rayj radium 
or radioisotope therapy o 

2o To escplain pertinent basic information about 
.- ■ . radiation in a simple and direct v&y<. 

3o To Bjake provision in this publication for 
yearly supplessents or replaeen^nts as needed 



268 



CeseiKlIATICl? SSESU ° 1 serial n©c 



J£^s2M^ 



An advisory (saamitt®© (as notad above) is working with 
tb@ principal i2ivsstlga-fe©ys to help deteymiia® wbat inforsoatisn 
is mo3t neeticKi asid how this can best be presented.. This 
gr®^p is c^isulttne with ns^Lical (particularly radiologists} 
and nursing staffs in ©ach institution represented o It is 
plesHoed that written materials will be submittM to the group 
by those having special interest in certain ar@as; these will 
bs reviewed and ineorpostited in the manual by the principal 
investigator o lUustrativ© material frs® th© various centers 
is also to be sifcadtted for rsvisw and inelusion as dis®md 
advisable o 

SigBificaaee: 

There is unanixoous fe©ling as^toQ the advisory group 
that there is a definite need for such a publication and that 
it should be smds available as socn as possible » Several of 
the physicians assoelatsd with the nurses on the ceeBaittee 
have also escpressed their interest in seeing this publieation 
coBspleted la the near future » 

Prgyposed CQigse; 

Cosi^letion of the mawaal aM dlsseaiinaticai to nursing 
personnel concerned (l) with the direct care of patients aad 
(2) the instruction of those who are providing patient csre in our 
hospitals o 



Part B included Yes /^ Ko ^ 

269 



Serial Hq . MCI°1£^ 
lo Fi«M IsKvestigations aocL 
PHS^NIH DemoiaBteations Braaeh 

Individual Pr^jeet Report 2o Nursing Saetioa 

Calendar Year 1959 3= Silv«r Spring, ifajylaad 



Part Ao 



Project Title: Investigation of Problems (Mental aad Physical] 
Faced by Patients Itoiergoing Tr^ataent for 
Cancer at R®swell Park tfeaaoriej. Institute ^ 

Principal Investigators Earlene Fiimerty 

Other Investigators; Rosalie lo Peterson 

Cooperating ISaits: None 

Ifen Years (Calendar Year 1959 }s 
Total: 5/6 
Professional J 5/6 

Other t Nooe 



A pilot stidy t® test the feasibility of studying post 
hospitalized caacer patients for a specified period of time 
in order to find out seise of the major probl^as faced by the 
patient and his fasdlyo 

Clbjectives: 

lo To learn more about the fears and anxieties of cancer 
patients and the problems which may delay or detract from the 
fullest rehabilitation of cancer patients o 

2o To ascertain the extent to vbich nursing service 
may help meet such probl^ns^ 

Methods Enrplpyed: 

Data will be collected through home visits and clinic 
interviews o 



It is felt that a sttdy of this kind will point up 
certain information which may help nursing leaders to prepare 
student nvffses to meet the problems of individual cancer patients 
and to provide sv^port to the patient at the tine it is most neededc 

Proposed Course: 

To design axid execute a more refined reseaireh project 
after the exploratory phase is ccoipleted in 1959 » 



Bart B included Yes £7 ^° S 



caHPHfUASIOT SHEST ■= 1 Serial No» mi^lBB 



Methcds Baplgyed; 

lo Plans ,f®r cojiferejaees vere made directly with each 
State league for Nurgingo 

2« Timg alloeat@d t® each institute was limited to 
one day, although ia larg^ states tvo las' msge) separate 
iostitxztes -were S'Sasrtiiags coMuetedo 

3 a The cuEs&er of participants vas restricted, in so 
far a$ possible ^ t& 50 "but oa. oceasion there wer« 100=200 in 
the g^^upo 

States in Vjhich C ^^ueted; 

Mzm^sota CkXahotna 

WycBdjag (2) North Dakota 

PecnsylvBaia New York (2) 

Colcsado Rhode Island 

Signifieance to the Program of the. jnstitute g 

These eonferemees "brought together f&ciities ftfOTi 
several schools of nursing (or in seme instanees faculty of 
a single instltuticax) in aa alasospher^ which promoted critical 
analysis of how mueh en^jhasis is actually being put upon 
furthering gpcd inter^personal relationships between nurses 
and cancer patients j, azad how much emphasis is being placed on 
the development of probl@n= solving skills among ninrsing 
students o Several groups have been stimulated to schedule 
additional meetissigs asA/c^ faculty discussion groups to 
©sntinus the analysis begun during the confteren^ periodc 
Ultis^tely this kind of analysis should lead to iisproved 
teachings and in tumj to better care for cancer patients « 

Proposed Course of the Project ; 

To disccatiBi^ conferences since it is felt that 
faculty in the schools of nvErsing have had sample opportunity to 
beccaae acquaintisd with these teaching tools; to eoastinue offer- 
ing the testing matsrisJ-s to schools of nursing on a loan basis c 



Part B iaeluded Yes ^ No ^ 

271 



Serial N©a HCZ=>aB8 

lo FislS iBvestigBtions aasL 



lodividual Px^Jeet Report 2o Kuyalcg Seetioa 

Caleafias' Yea? X939 3» Silvsy ^rixsgi, S&xylaad 



Part Ac 



Project Title: Work eosferenses {&a Nm'se Patient Relations 
ajffid Problsa Solving lests) through State 
Leagues for Kus'siago 

P?ii3cijal Investigators: Elizabeth Walke? aed Rosali® lo P@t©rs©® 

Other Investigators: fia^m 

Coc^rating Units: Nosae 

Ifen Yeas-s (Calendar Year 1959): 
Stetal: lA 

I^fessiesjal: l/ii. 
Crttier: l/l2 

PTDjeet Eescription: 

Objectives ■=» Ultiaaitg ; To improve nursing care to 

caaeer patients <> 

(a) To test feasibility of 
introdueing tests to large 
groups of instructors at ons 
tims instead of vorldbag 
separately with the faculty 
in each school of nursing c 

(b) IT© detessaine whether cae day 
would suffice {prtviously the 
prsgraa had been 2»3 days)o 

(c) T9 detemine tdiether futv>r® 
eonfierences ©oiiia be initiated 
and @arri@d out locally with= 
out further assistaaeg frcas 
this office o 



272 



Serial Ha. ^S-193 
lo Fieid lavestisatisQS aood 
PHS^HIH DsuEmsteBtioas Branch 

Individual Project Report £» Nursing Section 

Calendar Year 1959 3o Silver Spring, Maryland 



Pert Ac 



Project Title: Appointagot to C^isaitte® on the "Role aM 
Training" of Professional P®rEonnel°=White 
Hovss© CisQf©renc@ ob Aging=.=196l, 

Principal Investigator: Rssalie lo P@t©rsoe 

Otbsr Investigators: Nuraing Coasyltantsj, Chronic Disease 

Branchy Bureau ©f State S^rviees, USPHS 

Cooperating Ifisitss Karae 

&^n Years CCaleaaar Year 19591- 
(total: 1/12 
Professional: l/l2 

Other: Hone 

PrQject Description: 

Th.® Egjafcers of the Public Health Servic® to serve on 
eosxdt'^ss of the Whit© Bsust Conference on Aging w®r® 
designated by the Of fie© of ths Surgeon General o 

MMsbers froBo, each profeesiosal disciplins v@r@ asksd 
to prepare a statenjent of the philosophy of their pacofessional 
gi'ovtp and a stat@3s©nt of the philosophy of the Federal 
Goverai^at in regard to professional ne^ls and traiaaing 
requires^nts f relating t© "aging").. This stateaeat of the 
philosophy of the nurses in the USPHS and of thosse in other 
Federal Departesnts has betn coaspleted and oeeds for nursing 
service aM nxsrsing ^iscaticm noar and in the future have 
been projected*. 

This portisss of -Uti® project is completed o 



Part B iaeluded Yes ^ No ^ 

273 



Serial No a Nei^=19^ 



lo Field IssvestlgatiiDns aod 

PHS=HIH Demonatrationa Branch 

IndividiMil Project Repeat 2, Nursing Section 

Caleodar Y©ar 1959 3- Silv«g> Spring* MaxylsBd 



Stert Ac 

Project Title: Attitudes Test (AliQut Cancer Nursing) 
Principal Investigator: Lily Co Sagen^n 

Ofcher Investigators: ISum 

Cooperating Itoits: Cancer Control Section^ 

Special Health Services j, BSS 

MBSi Years (Calejadar Year 1959): 
!Bo'bal: 7/U? 
Professioxffil: 6/l2 
Other: l/l2 




lo To design a written test which will assess nurses' 
attittides about cancer aaad the nre-sing care of patients 
who have cancers . 

2o To sffike this test available to instructors in 
schools of nursing azsd ottters responsible for teaching 
the care of mincer patients 



lo Review of the literature for existing tests and 
for references relating to the preparation of attitudes 
tests o 

2o Coosuitatioa with psychiatrists^ psychologists 
and other experts re formulatioa of test questions . 

3o Construction of test itemsj questions, etCo 

ko Trials of the test questisas with nurses, 

lastruetorsi, etc, RefiDeaent on basis of suggestions d 

Propoeed Cgigs®; To coocplete this t«st in i960 and t© 
validate it for lise with i®ay.groi5)s of nurses, io e- basic students, 

JSixirsss isi hospitals j> public health nurses ^ etCo 

Jart B inslwded Yes ^ Ho ^ 27^ 



serial Uoo »CE°a95 ' 
lo Flmli. Isveettgationis azsd 
PBS=NM DffiB©©8ti^ti©iis Branch 

Iaaivi^t;ai jPr®jc«t Report 2o Nu£>8ing Sacti®n 

CaleMar Y®ay 1959 3o Silv«r Spjrlags tfeiryland 



Paart A- 



Pr^Jeet Title: Stufly of Metl^edolosy far OsjKluctiBg Nursing 
Research Ps-ojectSo 

P^lneipal ajvestigstor; Lily Co Bagearmaa 

CrtSxer InvestlgiBtars : Elizabeth Walker, Josephine Pateraoja^ 

Sarleae FlEserty aM Patricia Get©er 

Coop^^atias Iteitg': None 

Mas years CCalsMe? Year 1959 )s 
T©tal: 1 1/2 

Professioaal: 1 l/2 
Other; Bmxs 

Project BeecrlptiQs; 

1» Reviewing the liters tvire aa. RuFsiag Researeho 

2o SxploriBg vsriodis mirslng problems in which resedreh 

is needed » 

3<. Practice in setting up research designs far specific 
projects throiagh group thinJcing ai^ plansingo 

ho Cosisultati®z]i@ with various experts is other fields: 

£° 2?^ ^ifi ,, I CsBsultants, Research Nursing 
DTc Play® AbdeUah } Division Hursing Rt&ources 
DTo Ls^»rd lo Pearlin Psychiatrist - HJME 
Mrso Helen Tibbitts Research Assistant j, 

Divisiisa of Research Grants 

5o Study ©f various technlQues fi9r obtaining data fr<am 
individuals and groups « 



This project is csaspletedo 



Part B iscluded ^©^2^7 ^® S 

27. 



Serial N©o K0I'=^196 
1 o Fleld~^jvesti^ticms and 
PHS^RIH Denenstyations Braneh 

Individual I^^j«et Report 2o Nursiog Sectissn 

CaleiKlar Year 1959 3<. Silver Spricg^ Marylaad 



Part Ao 



Project Title; An ejqjloifatisE of the feasibility of nsaking a 

©saaxpariso© of the incideace of eascer and other 
diseases in a selected group with the incidjsnee 

of those saa® diseases ia the geiseifal popvilatioj3o 

Principal Inv^stigBtsr:. Patricia Bo G®is®r 

Other Investigator: DTd Al^gsander Go Gilliam 

Cooperating IMitss None 

Han Years (Calei^aar Year 1959); 
Total: 1/6 
Professioisals 1/6 



Objectives; 

lo To calcijlate the e:2pected and aetual incidence 
of cancer aod other diseases in a snail ^ closed popvilaticss 
of professional vcgaeno 

2o To eoB^are the findiasgs of this groxrp with 
other conparable population groups » 



lo Einplc^ the actuarial method of calculating the 
expected incidence of cancer exjA other diseases in a 
selected pqpulatiaa group o 

2o Determine the actual incidence of these 
diseases in this population o 

3o Deterjaine the si^iiflcance of a costtparison of 
these findings o 

Proposed Course of Action; 

If the findings are significant ^ to ccsipare the 
origii^l group with other similar ^oups in the ^neral 
pop\jlation-> 



Part B inelMed Yes ^ No ^/ 

276 



PES=N2H 
Bidlvidual Pr^jeet Report 
Galezidas' Year 19^9 



Serial HO c HSZ^Ag? 
lo Flelii finrestigatloois and 

Demsnstratlcms Branch 
2. ]9ureing Section 
3o Siiv«y Springs JbJ^land 



Part Ac 



Pr-s^ect Title: Coasultatlon to vuaiversltles interested in 
specialized pro-ams in Can@er Kitrsiiig 
(Baecalaureate and graduate pro-ams )o 

I^iBcipal Investigators; Rosalie !<, Peterson azii 

Elizabeth Walker 



None 



Caqperating Units; 



Ifen Years (CaleEdar Year 

Total ; 11/12 
Professional; 9/12 
Other; 2/l2 

Beseription: Several visits have l>ee& nade to New York 

Ifeiversity aiaa. Teachers College^ Colwibia Utoiversityj, to 
explofe the need for courses whieh offer nurses the 
opportunity fo^ special preparaticm in caneer nursing iu 
programs leading to ctasters and doctoral degrees <> 
Consultation has also heen carried on in several basic 
p3x>grafflS2, io eo von a haecalaureate levels, in such schools as 
Iteiversil^ of Wasbingtonj, University of Cregon^ and 
University of California » 



Proposed Coxgee; To continue 
interest in 
Qurslngo 



c©nsvd,tation as solicited and t@ further 
the clinical programs in cancer 



Part B included 



Yes^ 



No 



277 



Reeeerch Grants Branch Annual Bs^rt 

National Cencer Institute Calendar Tear 1^9 

Objectjyea 

'She objectives of the Research Granta Prograia of the Kational 
Cancer Inatitute nay be stated as tolXcmsi 

(a) to determine the agents and cosditione tb&t induce 
cancer and the raechaniszns through which they acto 

Cb} to elucidate aad analyze the factors that proiaote 
canoer developii^nt ao/d groifth after it has been 
inltiatedo 

(e) to characterise the various types of cancer as 
distinguished froia noniial aad benign groftrth, and 
the effects upon tJie individuals la \irtiora they grcsf 
by vhieh their presence, type and location can be 
identifiedo 

(d) to devise effective nssans to prevent the initiaticm 
of cancer,, to control its growth and to eradicate ito 

By providing flnsjacial support, the prograjn asaisted during 
the past year ©pproxiR»te3^ 1,500 sclentiste throughout the toited 
States and in aoase l6 foreign countries in their efforts to attain 
these objectives., The diverse j^thod©^ diaciplines^ and talents 
that ajre being brought to bear in research on cancer are illustrated 
by the description of reeear^ accon^lishaientB to follo*fo 

Plans and Froble aa 

Perhaps the question of plans and probleoffi should be presented 
after a discussion of ressearch acaooiplishsMmtSo Tbie question is 
discussed herOj, hove^erj. beceuBe of its general iinportance and because 
it is otherwise likely to be overshadowed by the simple oiegnitude 
and volume of the presentation on research accooiplishzaentSc Plane and 
problesia are presented together because they frequently ore closely 
relatedo 

As Cancer ^eaaotherapy Cooperative Qinieal Oroupo 

During the past several months the responsibility for the review 
of applications and the administration of grants concerned vith support 
of cancer d^motherapy cooperative clinical groups has been transferred 
to the Cancer Cti^ootherapy National Service Oentero Ibese groups con- 
sist of invest.'' gators in noa-Federal institubions who fomasd coopera- 
tive groups a+ the request or irith the assistance of the Csater primarily 
to provide elinical trials of compounds originating from the Center's 
progrcHa of producticm and sereeningo 



278 



- 2 - Annual Report 

RSB-UCXE 1959 

This di@£ge ^&s authorised by & ruling on the x>srt of the 
Office of the Director, Natiosial Institutes of Health, that appli- 
cations for support of «ork in vhich the staff would play a major 
9S^ continuing role in the deeigD azui conduction of the study could 
be revimred by a Special Heviev Conaoittee instead of by a Study 
Sectioi. 'Sas Diarector of the National (Smcer Institute requested 
that ajrplicatioas for support of cooperative groups be included in 
this category to be reriewed by the Clinical Studies Paaiel of the 
Center in place of the Cancer (Siemotherapy Sttai^ Section prior to 
review by the National Mvisory Cancer Council. The Council, at 
its rceetin^ in Hoveiaber, 1959* concurred in this arrangemwit with 
the laQderstanding that the oei&ership of the I^nel would be broadened 
to inclxjsde clinical investigators vtho were not themselves participating 
in a cooperative gronspo 

As a smtter of record end interest, existing cooperative groi^^s 
are as follc^m: 

Eastern Cooperative Group 
Acnite Le\ik@B£ta Group A 
Acute £(@ukefmia Group B 
Western Cooperative Qrowp 
South^uBtem Cooperative Qroitp 
Cooperative Breast (Suacer Group 
Siirgery Adjuvant Cooperative Gseaap 
Soutisifestem Cooperative Qroup 
Prostate Cemcer Coo;perative Group 
Breast &acer Surgery Adjuvant Group 
Ovarisoi Cancer Surgery Adjuvant Oro»q> 

!l^o sro\^s for ^iiaich applicaticas are pending are; 

Hidvest Cooperative Grovqp 
European Breast Cstncer Group 

Applications and grants for these groups Involve approxiniately 1^ 
investigators at 13^ institutiosae for a total of (^proximately 

$2,700,000o 

Bo ?iruses and Cancer 

During the past year the Cotmcil, with the assistsnee of its 
Viruses and C^icer Banel, the Virology and Rickettsiology Study 
Section, and staff, has encotsrsged a narked izierease in research on 
viruses in arelation to cmicer, particularly in relation to cancer 
in B«a» Sae folioalng table tiham the increase in activity in this 
field over the past three years o 



279 



- 3 - Aimual R«port 

RCffi-HCI 1959 

Fsr xw( ^ n 1958 n 1959 

Amount of Txm&a, Awarded fiyS^TS^ |2, 103^063 fe, 636], 975 
Hoo of GsE-ants ReconsDeiided 63 79 IO6 

It is aatleljt^ted that In FI i960 well over four nilUon 
dollars vill "be awarded for vork in this fieldo Btxls amount will 
include tvo larg®, speoial purpose grants 1 ooe to the South Jersey 
Medical Reoearch Fovrndatiom to establieh a ceottral cell line bank 
and regiatry to jsaint«dn, suk?!^ and stu«3y tissite cultijr® cell 
str&iBs of special Issportsxice in vsa*ioua research, and aoother great 
to the Araericau Ty^pe Oilture Collection to establish a long-term 
repository asad distribution center for v&pIcub tumor viruses and 
tissue culture cell llcsso At a recent losetii^ of the CJouncil^ the 
opinicm was eacpresised that there had been sufficient stimulation 
in this field for the tint® belngo 

Co Isolation-Psrfusion Studies 

Ikiring the past year there ha& been a noticeable increase 
in the number of applications requesting support for the applica- 
tion of isolation-perfUsioa techniques to the treatment of cancer o 
aiis technique inyolves the isolation of an organ, a lii^, or a 
siiecific area of the body, from the rest of the circulation and 
the pcirf^ision of this part of the body with chemotherapeutic agenteio 
She advssTfcage of this approach is tiat higher concentrations of 
the agents can be tf:>lerated thsai vbsa the agents are distributed 
throughout the patient o ISiese applicatioos have presented a problea 
for aeveral reason© e Many of them p3X)pose little nev except the \3se 
of a different agent or the application of this technique to a 
different orgaa or area of the bo<3yo Also, those applicaticsas ccm- 
cemed priicarily with surgical technics were reviewed by the Surgery 
Study Section and those application© using well developed surgical 
ffiettiods for the perfusicas of cheBsotherapeutic agents were reviewed 
by the OBacer CJberaotherapy Study Sectlan.. As a result, neither 
group saw the ccttsplete pictvtreo 

In an effort to resolve this problem a meeting was held 
including some ineznbere of each Study Sectiono It was pointed out 
that the Osmcer Institute is supporting only 10 projects in this 
field totalling a little more tl^ $150,000 snnuaUyo It was 
agreed that aiore good work in this area should be supportedo It 
was felt that &ay striking results would have to awB5.t the availa- 
bility of more effective agents, but that eontinuiSd research in this 
field is iir^rtaat so that when xaora effective agents are available 
"Kie surgical technics for their use 'alll already be in hand.. It 
was also agreed that requests for support for work in Idiis field 
shoxild be considered on their n^rits, and that proposals preseatSjsg 
no tmf sux'gical or chi8inK>th9rapeutlc aspects should iKst be reccasaendedo 



280 



- ^ - Annual Report 

ROB-KCI 1959 

Do Cooperative Studies in Radiation Therapy 

For the paet two years the E^'Hation Study Section has been 
giving serious conaideratiOQ to the posslbllilqr of developing one or 
jnore cooperative groups to evaluate such problems as x-ray therapy ■ 
under high o^gen tension, short-term and long-term effectiveness 
of comi-entional and eupervoltage x-rey therapy, and the eojaparative 
effectiveness of supervoltage, isegavoltage, and particle x-ray 
therapy against cancer.. With the aid of grants from the Institute, 
the Radiation Study Section has sponsored several conferences to 
consider this problooo As a restilt of a recent (January 8, i960) 
meeting, it seaais doubtful that it will be possible to establish 
in the imaodiate future truly cooperative groups of the kind that 
have been organised for cancer chemotherai^o 13ie consensus at the 
meeting v&s that codes would be developed to permit uniform reporting 
of radiothsrapeutie effects on cancer of seven different sites » 
After acquiring end analysing preliminary data from this source, 
consideration will be give» to such further dsvelc^pments as nay ©i^m 
desirableo 

Eo Use of Research Oram; Funds for Foreign Travel 

It has been about one year since the requir^Bent was eliminated 
that principal investigators had to request persaiseion frcsa the Riblic 
Health Service to use researdi greot ftmds for foreign travelo Eie 
I>olicy is now that grant funds iaay be used for this puipose if the 
grantee institutioa determines that the travel is necessary to the 
successful prosecution of the projecto The only remaining reqiiiremeat 
is that all foreign travel be reported in the manual suEEisary progress 
reporto 

l^en this chasige was made, some concern was ejcpressed that this 
privilege would be 6bt»edo Perhaps it is ^K>3rt;h noting here tiat this 
office has seen no evld^snce that such abuses have occuxredo 

Fo Reviar of Foreign Applications 

She review of applications frcaa institutions outside of the 
United States continues to be a problan., The auniber of these appli- 
cations is increasing, apparently pertly due to mora frequent visits 
to foreign countries by ccmsultMits and members of the staff sad 
partly because of a widely disseminated impression that the Uaited 
States is interested in supportiag aaedical research abroad* This 
irapressiOQ may have resulted in part from discussions about en Insti- 
tute for tatezTtational Eealtho Frequently foreign applicaticaas 
provide very inadequate infoxnaatioao Siis lasy be due to several 
obvious reasons such as a lack of facility with -yie aiglish laE^uage 
and a lack of understandia^ of the kiJJd of inforj^tlcm that is desiredo 
A revisaai application forru mist baas recently bec<M2» available may 



9R^ 



- 5 - Annvjal Report 

RGB-RCI 1959 

eHminato part of this problenjo It is felt, however, that further 
c<m8ideratico should be given to providing special instructions to 
foreign applicants to aid them in providing the necessary infomaationo 

Oftentluss a consiiltant of the Public Health Service vill 
be for other purposes in a countiy or in a nearby country fran vhldh 
foreign applications are praidingo It would be most helpful to have 
the confiulteat visit the applicant., Apparently, however, the pro- 
vision of the usual expenses and consultant's fee for this purpose 
is prohibited even thoxjgh the cost might be less than for a project 
site visit in the Imited States <> It is tjLnderstandable that consultants 
cannot be sent all over the world for site visits. However, when a 
ccajsultant vlll be in the vicinity of a foreign applicant, it seems 
unfortunate that ireasonable additional expenses cannot be provided 
for a visit o 

G, Study Section Prioritlea and Approval Rates 

During the past year the Council, with the assistance of the 
staff, has made a thorough review of the priorities and approval 
rates of all (22) Study Sections that reviewed "C" applications » 
This stiidy included not only "C applications, but all applications 
reviewed by these Study Sections at all three meetings during the 
past yearo It vas found that 26ol^ of the applications vere dis- 
approved, 55 9^ of the requested funds vere reccanmended, and the 
average priority vas 2U2o In general. Study Sections in the so-called 
"exact" sciences like Biochaaistry, I^alologlcal Gheaiistry, and 
Biopfcorsica and Biophysical {3ienjlstry, have better than average prior- 
ities (205-227) and lower than average disapproval rates Cl2o8^17o03S)<, 
Shis is not unexpected since it probably is true that applications in 
these fields are better than average because the applicants are better 
esnd more highly trained for research than in fields like some of the 
dllnlcal areast, for exaia£>leo 

In general Study Sections in the clinical and more applied 
areas like Radiation, Heurology, and Cancer Caiemotherapy had poorer 
priorities (253-282) and higher disapproval rates C32o6^-4l„l^)o 
!Rfo Study Sections differed from this generaliaatlono Oeneral Msdicine 
had an average priority (2^1)/ tiut had the highest disapproval rate of 
all (U3o25&)o "Hie Pathology SttuJy Section had next to the poorest 
priority (279), but had one of the lowest disapproval rates (16. 6^) c 
catnicians have felt that it vas difficult for them -to obtain support 
because the basic scientists among the reviewers were not synipathetic 
to clinical problems o !Hie above figures indicate that this is not 
the case, but rather that ^en elljaical proposals are reviewed by 
clinicians, the approval rates are still relatively loWo This may bo 
due to the fact that although the applicants may ba good clinicians, 
they freqxiently have had only limited research training and experience., 



282 



- 6 - Anaual Report 

RGB-HCI 1959 

Besearch AcccaqpllBfaBgats 

Aa Oetfcinogeaesls 

"Bie process of carcinogeneBls or c^acer lEKJuctlon me^ be 
brausbt altotit "by playsic^, ebemleal, or biological oeaxis if ooe 
egrees that viruses are biological rather tten ehesaicalo a3se 
role of viruses in careinogeaasis is recaiviaag a great deed of 
attention,. Since tl2e first report of the induction of cancer by 
cell-free filtrates in 1911^ viruses have been shown to induce " 
tumorfi in chickens, phs8«snts, ducks and other fowl , mice, frogs, 
rabbits and other gmisBBls^ As indicated ^irlier in this repoirt, 
the Institute has stiisulated increased research activity in this 
fieldo To date, however, there has been no direct evidence that 
viruses plsy a role in carcinogenesis in wmo 

Work continues on tl^ characterizati^i of the msasaary 
tmcor Mlk agent in mlcCc Biological procedxirea have been used 
by Dra Item Ho Moore and bis eoll^sgues of the professional research 
staffs of three institutions; 'The Rockefeller l^titute. Tins College 
of Hayaicians end Siirgcons of Ctolucfcia Itolversity, and Xale Itoiver- 
sity, to determine the size and strticture of the biologicalljr active 
agent responsible for the trsnsjsission throu^ ssilk of mouse wamavcy 
cancer, fSie filtration of jailk froia high-breast-cancer mice <tf the 
R XIX strain through gradocol m&sbv@zme vith decreasing pore sisee 
indicated that a minisnsB of activity passed through interoiediate 
pore sizes (ICX) to l60 Milliaicra) o Filtrates throu^ sjaaller porea 
were aigjaificaiitly activca Mlk treated vith siuall z-adlation doses 
of bes»7 hydrogen nuclei produced oore tumors than the control^ un- 
irradiated sdlk. larger doses indicated a particle size much less 
them 100 loill^n^cra. Free diffusion esporiisents ii^Ucated that the 
activity was associated with particles of two different sizes., Al- 
together the data denoted the presence of a large agent about 100 
laiUiMcra in diameter and a snail agent 20 to 30 mLllimicra in 
dlaEseter or posaiJ>ly sia&ller<. Furtheraore, the presence in the Bii3Jk 
of an inhibitor of size ^40 tO 60 aillliiBicra is Indicated by the 
results of all three approaches » Tbe complex nature of the aiilk 
egent disclosed by the physical Qseaasureasents agrees with the picture 
of one of the structures revealed by electron microscopy aa well aa 
with seemingly conflicting i^asurea^ats reported in the literatureo 
Sbe large agait defiaaed by •tiiese indirect methods corresponds to 
the whole particle seen in the electron siicroscope and the sraall 
agent corresponds to its internal core or nudeoida l&iieae researchers 
suggest that the nucleoid is essentially a nucleic acid which Hsay, 
in the absence of the " inhibitor" # retain its activity after having 
been stripped of its outer laenferane or sac 



283 



- 7 - Annual Report 

RGB-NCI 1959 

The Inherited susceptibility for the development of spontaneous 
msmmry cancer has been found by JJTo John Jo Bittner of the University 
of Minnesota Medical School to be transmitted equally well by both 
male and feaale Mce of cancerous stock. In one cross, linkage was 
denionstrated between the heredity factor producing brown coat color 
aud one of the caacer-susceptibility factors o These factors influence 
the production of traits in the aniasal as indicated by the naioes given 
to themD While the frsq,u0icy of the linkage may be reduced with the 
elimination of the mswmxy tumor agent, this does not alter the in- 
herited susceptibility, and the incidence may be restored by the admin- 
istration of the agent » The genetic make-up of animals of an inbred 
strain for the genesis of spontaneous maisnary cancer may be determined 
only by testing mice that possess the memmxy txaaor agent and have 
been subjected to hornional stimulation adequate for the induction of 
these tuEiorSo It has not been ascertained if the inherited suscepti- 
bility for the g«5nesis of the usual type of spontaneous msmnstry cancer 
Tt^ be similar to that required for the genesis of mammxy tumors in 
agent-free anJjjals including mammary tumors induced by cancer- causing 
agents o Several inherited hormonal patterns were found: one of these 
plays a role in the Induction of msEmry tumors in virgin females, 
while another may inhibit their develoiH!®nt in breeding females of 
susceptible stocks with the agento Preliminary data indicate that the 
inducing pattern may be dominant over the inhibitory mechai^ism, although 
the latter mty ^jert some influence over the average cancer agCo Observa- 
tions obtained in several hybrid crosses stiggest that the same hereditary 
factors do not control the inherited susceptibility for mammary csuacer 
and the wsmmxy cancer-inducing inherited honsonal influence, for the 
inducing hormonal pattern jmy be transMtted by mice that are resistant 
to the developanent of EKmmary cancer. In adclition, mice that are sus- 
ceptible for the disease may transmit the inhibitoiy ho3:Taoned influence, 
and this tends to mask the inherited susceptibility<. 

X^ta have been obtained during this study showing that the in- 
cidence of spontasieous wesssasy cancer amy be no higher in breeding 
susceptible females havlsjg the mammary tumor agent then has been ob- 
served in ageat-free susceptible breeders, from which the agent eaoaot 
be ren»ved by biological assay., Follo?ricg the introduction of the 
ffiansnary tumor agent into agent-free feaales hy such methods as nursing, 
injection and male transmission, the progeny of such feEBales will have 
an increased incidence and acceleration in the time of appem*ance of the 
tumors, and the agent can be demonstrated to be present in the tumors 
from the Infected sKtthers and their offspring. Susceptible females affiy 
not show similar incidences and average cancer ages after they have been 
nursed by females of different cancax'x>us stocks o It is not known whe-bher 
the sensitivity of agent-free females of various strains to infection 
with the agent from males of cancerous strains laay be influenced by 
another genetic eoEg)l«X4 



284 



- 8 - Annual Report 

RCEB-HCI 1959 

A great dsel of work is being done on the viral induction 
of leiikemia in aniioals and the related leukosis in fovlo 

An eeent has bean described by Drs.. Ro Po Buffet, S<. L. 
Cans33erford, Jo Purth, and Md Jo Hunter of the CMldren's CSancer 
Reseaoreh Povmc^tion and the Harvard Medical School, and ^diich 
csAoses tumoroxjs lesions in AKR, Rf and C3H ndcCo Cells of leukaaia 
^rere obtained frcsa two seven-month-old AKR female jalce, each vith 
spontaneous tlayaaic mjd generalized lyagshoBja after tliiree serial 
auocessfUl subpassageso Fresh leukemic tissues were passed throxagh 
a SO-HJeah steel screen and suspended in a buffered solution.) Sie 
supernatant fluid derived txxm centrifugation of AK leukeinic tissues 
at 105,(XX)x gravity caused a Eultiplieity of ttenors and a variety of 
nontuimrous lesions to &J«^ of AKR mice injected shortly after 
birtho ®is evidsace reported, both by these investigators and by 
otlierS;, sugges>5S that 'the lesions found under these circumstances 
are caused by a plu;ripotent agent (to be called F virus) vhich is 
distinct frcaa that iihich causes leukeaia (to be called L virus )o 
!Ehe relationship betsre^s P and L viruses i.8 conjecturalo to the 
basis of present evidence, it is suggested as a working hypothesis 
that the P virus i© of the parasitic type, is very small, and spreads 
by natural ineanso It is usually masked, and may be inhibited in 
tissue extracts by soine macroraolecuXar particles o "nie L viriis is 
larger, has a usore lijmited host range, and nssy be related to the 
hereditary structures Cscnoiae) of the leukeinic cell* (Dto Purth is 
ttov at Roawell Park ffemorial Institute )o 

Drs» H, Mo Schoolman, So Oe Schwartz, W, A, Spurrier, and 
Po B. Szanto of the Hektoen Institute for Medical Research ha\'e 
reported their experiences with leukemia of mice vhich was induced 
by a cell-free filtrate » A 22-%?eek-old C3Heb male rosuse was observed 
to have developed a spontaneous leukeinia<> Tumor cell suspensions trotn 
this eniraal grew readily when transplanted into other C3Heb mice^ 
The cell filtrate i^sed wat prepared froia its brain, and induced Isukeiaia 
in froa 12 to SO days in spproxiEately 70^ of adult animals ^ich had 
been injected via the brain-case or into the abdornin&l cavity., ^e 
induced leiikeiaia is characterised grossly by slight to lEodei^te enlarge- 
ment of the peripheral, and moderate to m^ked enlargen^nt of the 
white- cell lys^ glands in the center line of the chest c "Hie ttsymus 
gland lying Just above the heart was variably Involved, and a diffuse 
tumor of the aieiabrene-likc intestine-supporting mesentery extended into 
the tissues of the excretory and reproductive tracts <. Enlarged livers 
and spleens vrere alsousualls' presento Microscopically, massive ^jd 
diffuse iavolveiaent of the lymph nodes, liver, spleen, and urogenital 
org^is is seeao The lungs sho^' a diffuse infiltration of the alveolar 
pa2*tltion8o The bone sorrow and blood are inconstantly involvedo The 
process laay best be characterised as a lyinphoblastic lymphosarcoma, 
^id eaunot be differentiated from the 1 likeniia of AK miceo She filtrate 



285 



- 9 - AnnueO, Report 

RGB-HCI 1959 

is inactivated by heat (65 degrees for k^ miBUtes), retalne its 
activity after at least short periods of freezing, and is not 
inactivated by exposui^ to as rmxch as 100,000 roentgens delivered 
by radioactive cobalt 60 in I5 minutes o Both the tumor cell trwis- 
plant kM the filtrate have been shoyn to induce leulcemia in various 
sublines of the C3H strain, as well as the Fx hybrid of a C3H x 101 
crossingo Neither txinsor nor filtrate, howc^rer, produce a leuk«nia 
in Swiss or MR mice= 

The electron microscopic app^uraace of tumor cells in birds 
has been studied by Drso L» Dtsrachowslsl, C Eo Qrey, Bo Ro Burnfflster, 
and Wo Oo Halter of the Itoiversity of Texas Mo D» Anderson Hospital 
and Tuiaor Institute, the Baylor Iftiiversity College of I>4adicine and 
the Uc S» Departaieat of Agriculture Regional Poultry Laboratory at 
East Lansing, Michigeco (Their investigations were a continuation 
of those on viral agents involved in the origin of different fowns 
of (rfiicken leukosis cou^jlex, knovn to be transmitted by cell-free 
preparations Bae present report concerned the examination of the 
effected org^s of chickeas with graauloblastosis (layeloblastosis) 
i^ei«iB there are abnorii©! nuitnbers of iMssatura bone laarrow cells 
paresent in the tissues <> Sssorous spleens and livers from chickens 
with grEU3uloblasto3is vere studied in ultrathin sections » In the 
cytoplasm of the affected cells chaages vere observed siaiilar to 
those seen in erythroblastosis, another sseafcer of the fowl leukosis 
coBiplesin Particles ^©re observed in the cytoplaoia of cells of siiuilar 
sise esi& structure to that of particles in erythroblastosis Average 
total dieaster of the particles vas found to be 67O A^ eaad that of 
the central dense core tfas ^0 A^o lo particles of similar sise or 
structvJre were observed in ultrathin sections of organs from healthy 
young chickens of siasilai? broedo 

ISae two projects described below illustrate the kinds of work 
being done in chensieal carcinogenesis o Urethacshas been long known 
to cause cancer, and for a considerable time it was considered to be 
tissue-specificj it caused lung cancero Recently Bra, Albert ITarmenbeMisa 
and Herbert Silveratone of the Mohasl Reese Hospital (Chicago) conducted 
research which indicates that urethK:eis a Eultipotential cancer- caiising 
agent, eej>sble of inducing or sugjsenting the fomation of varied tuaors 
in a niffiiber of different aniioalSo Long-tena application of a solutic© 
of 20^ urethaaeln acetone was aads to the skin between the shoulder 
blades of JMk and C3H luices and hybrids of a cross between female C57H. 
and mala C3H laiceo IMs chronic treatment was continued for periods 
ranging up to I8 laonths and total amounts ofurethaaeas high as I80O 
milligrams per mouse » Uretharedid not evoke skin tumors at the site of 
applicatioae However, five separate sad distinct lesions were induced 
or potentiated by the long-term application of tarethiaa to the skin of 
the mice: puLiaonsry cmicer, asaasaary eaaeer, aalignaat cwseschy^^ tvusars 
in the f&t associated with the shoulder blade, tuniors of the tear glend 
sxaS. blood cysts in the liver» It is concluded that iirethas® is a malti- 
potential cause of cancer.. 

286 



- 10 - Annual Report 

affi-Nd 1959 

She cor^inatioj of cancer-causing derivatives of benaacridlne 
vith protsiniB h&& been investigated by Pascallne Daudfil, Osbriel 
Valie and Rene Vasques of the lastitut Pesteui^. Upon the aHslicatlon 
of 2f 10-diEaeti^l 7ji S-becsacridine (msadfi radioactive by combining 
carbon"lit into its ajolecul^r stiiicture) to the skin of mice, this 
very cancer- causing (xsmowaA ccciJined with cellular proteins apprcsd.- 
lintely 10 tia^s EKjre, auantitativ«ly, than 2, lO-diinethyl-5, 6-b«naa- 
cridinSi. Eae latter coaipomid has ^^ittle cancer- causing properties c 

Studies of the cerciaogenecity of tobacco tars continue to 
get attcntioa» Brsc Adele Bo Croninger end Evarts Ao Graham of the 
Washington ISaiveraity School of Medicine and Dr<, Ernest Lo W^der 
of the Slosja-Kstteriag Institute of' C^icer Research have coapaoped 
the biologic activity of different types of tobacco products such as 
cigax'S, pipes, and all-tdbasco cigarettes » !Efeey emplq/ed s smoKing 
inacbine therein a pojailar brand of cigars ■sas puffed three times a 
ainute, pijlll»g a. 50 nilliliter puff ever a period of four secosadso 
All-tobacco cigarettes ¥ere prepared by W2»pping cigarette tobacco 
in tobacco leaf » ISiese special eigerettes ijere snjoked under the 
saaoe coaditioaa as ordinary cigai^tte®, but, owing to the Tiraaty 
tobacco outer m-applng, th^ t^ere smoked B»re sloviy than the ordinary 
kind;, averaging l6|- loinut-es per eigarette<. A popular brand of pipe 
tobacco ^rsB ©jEoked on an appsaratus vhich pulled & 30 milliliter puff 
through each pipe every tea seconds j the daratic© of the puff vas one 
second* Bis resultant tars vere applied to the shaved backs of female 
G^ S5ice» ^e results suggest a somswhat higher degree of caacer- 
stimulating Ectivity for cigar and pipe tars than for cigarette tar 
and a scas^tohat lesser activity for all-tobacco ci^irett© tar coaipared 
vith standard cigarette taro Sobjs of the difference could be a result 
of differences in biological variatioa<, A coaparlsoa of \daole tobacco 
tar ^th diluted tobacco tar su^esta that dilution of tar accelerates 
tUEJor forssatlon, apparently «s a consequence of increased el>sorptioa 
of tobacco cancer- inducing imaterialSo A ccarparison of the cancer- 
causing activity of freshly pi«pared tar vith that of "old" cigaarette 
tar sho^'ed ao differences in activity. Sie present data re-eogjhasize 
the viewpoint that tobacco carcinogens are forised during the process 
of costoustion of tobacco and that the aasount of tobacco eancer-eausiag 
jmterials foiiaed depends upon the temperature reached and the eoaiplet©- 
nass tJith vhich the eoEibustlon process occurs. 'Sie results of current 
esperiai^ats are consistent vith human epidaaiological findings » ISiess 
scientists state that preventive aaeasures directed toward reducing 
tobacco cander^eauslng ssaterials nsjst apply to all tobacco productSo 

ISie opinic^ has been ©stressed that restrictions ^pon the avail- 
ebilil^ of osygea supply "ffill contribute eignificantly to sn increase 
in eaacero So tsst certain aspects of this vie^point;> Br.. Joseph A, 
Dipaolo of the Koswell Bark Jfesorial Institute has studied the influence 



287 



~ 11 - Annual Report 

RGB-HCI 1959 

of alteration in the esssunt of atmospharic ocsygen available to mice 
vith lung tuiaors induced "by the coH5)Ound urethane. dree groups of 
Hsale and feinale laice of the Strong A strain, 6 to 8 weeks old, vrere 
injected ^th cither OoOl milligrasa, or loO ndlllgram of urethane 
per graia of tc^ weight. SaniediateS^ after injection, mice from each 
of the 3 groigjs vere exposed for k8 hours to h different emriroaments : 
10 par cent osygea, room air, 70 p®r cent oxygen, and compressed air» 
Sirse EKjnths later the Mice were killed and the nuctoer of Itmg cancers 
counted* With 1 jsilligraja of urethane per gram of body weight (the 
ccncentzution ^Jliich produced tuiaors in all ssice) the group recelvijag 
70^ ojsygea (or asi egcgss ov^sr that ia the atiapephere) developed loore 
tuiEora than the other groxQJs, aad showed a statistically signific^it 
difference from the o'J^ier groups., 

Bo Sumor-Host Relationships 

The prdblaa of gro^^lng husaa tissue, normal and jaallgnantj, in 
aaiasslSj or of growing tissues from one kind of aoisjal in eaother 
kind Is an iBgKjrtsat oa® in csaicer research o Cte-owing hiai^n tissue 
in aaismla E^kas it possible to test readily various anti-cancer 
compounds directly ijpon hxnan tissusso IMs nsay be of sosise advantage 
although huimn tissues in aniisals are not the saiae as hissan tissues 
in their noras.1 hosto Saperisieats of this kind also contribute a 
great deal of isEamologj.cal infozisation aad will assist in understanding 
the relationship of the host to both traosplanted and spontaneous cancer*. 

^e sibility of csmcer tissue to grow, after transplantation into 
a new host of the saajs species, ia <lspendsnt upon the similarity or 
identity of the heredity factors present in the donor and recipient 
anisBlSo ©xe accepteibility of the "foreign" tissue, or its hlstoccsa- 
patibility, is an eagjression of these heredity factors. Dr<. George Do 
Snell of the Bosooe B« Jackson Meniorlal Laboratory has analyzed the 
production and typisig of histo-CQEgjatibility genes in the moufle, with 
particular reference to ajoioals ^daich have identical sets of genes 
(isogenic) or which have different espressions at the site ixi the cell 
where a given heredity factor is located (co- isogenic). Bairty-eight 
lines of Mica reached a stage of development where significant infonsatioa 
as to theJx refearable genetic eicpreasion (genotype) could be obteinedj 
26 of these were studied ia eosje detail. Of the 30 original isogenic 
and "resis-fcant" lines, 30 were shofaa to be distinguished from their 
inbred or colsogeaic imte by a difference at the H~2 locus. On© co- 
isogeaic pair, C3H &a&. C310C di^ered at H-1; wiother pair C57B1/10 and 
BlOaLP, at H-3« Of the 6 res^-ning lines, 1 prob^ly had an H-1 differ- 
ence, 3 were uucertain and 2, still under test, may serve to define new 
loci- Bvidenee was found idjat flx-st geasratioo hybrids, wi.th a tussor 
donor as one parent, are less ccsjpatible wl-bh tumor grafts than the 
native stjrain.. 



288 



« 12 - Annual Report 

RCS-Nd 1959 

Sbfi i^jysiologlcal fiactors in host-tusaor relationahlpa and in 
cancer disseMnaticai (lasBtastases) have been Inveatigated by Dto 
Ronsaii Molcaiait of the Woldemso: Medical Research Foundation, 3Sie use 
of a uell-sstablialied csacer graft as a transplant into norcjally 
r«oistaat hosts offers an expsrin^ntal situation suitable for study, 
fiinca the uKsltered live waacer cells csus be grafted into a func- 
tionally altered host. In his studies ha has investigated host 
alteration by maaas of active end passire iisaiinissation of the recip- 
ient hosts to cancer transplantation « Ttxe tumor used was Sarcosaa I, 
lethal to strain A jaice; it does not sxurvive in C57BL/6ja3c aicco 
HCTi5«ver, ^ea C57SL/6j'a:^ aica are passively iaamaxized with antisera 
to Sarcor^ I, end subsequently grafted i?ith live Sarccma I cells, 
the tumor gro^s progressively^ and the laice die^ A similar effect 
results fto® active iasmmizatic^ with frozen-^iried Sarcoaa I tissue » 
ISius, alteration by isssaronization of a nonaEuLly resistant host will 
result in cheages in ftznetlon, profound enough to alter the host-tumor 
relationship o In another series of studies with Sarccsaa I, C57BL/Jax 
mice were actively laaajnised with fitjzen-dried Sarccsna I tissue while 
sinailtaE^ously being treated with cortisone. Sie ecperlBantal observa- 
tions cited indicate that an altered host will"perinit" live cancer 
cells to survive. She ustastatic c^icer from the cortisone-treated 
and actively-iEEBuniaed CJTBL/^sJ'as mice was transplanted directly to 
norssal Eiice of six genetically unrelated strains, in which the "A' 
strain Sarcoasa, 1 does not norsally survive » In each instance, the 
cancer graft fi"oia the treated mice survived, gr«w progressively, and 
the mice died with large primary tumors at the site of lJis>laatationo 
DiPo Moloaut finds, hoKever^ that the loss of strain specificity of a 
live cmieer graft will occur la nsice without cortisone treatjianta 
C^JKi/Jax jDiee which had been actively iinsunized with froaen-dried 
Sarconsa I tissues veve iBiplsaited with a live Sarcoma I graft wMch 
grew progressively <, Prior to death, this graft was ti'ansplanted to 
normal untreated mice of the C3H strain, and is currently being carried 
by successive ti^ospl^itaticm and is now in its 50th serial tr^isplaait 
in norngd C3H asice« However, in C57BL/6 strain the tvaaor becajoa 
necrotic in the l^i-tb passage end regressed, Siis suddea rs^rassion aay 
have be^i due to esctz-aneous factors, such as bacterial cc»ita32iinati(»Xa 

The effect of repeated brief stress on growth of the free-cell 
Ehrlich ascites ((^^ainasm.) in the csouse has been investigated by 
J2t, James T« March, Bfe-o Bryce So Miller, and Br, Baldwin Go Laisson 
of the Dtaiverslty of California School of Medicine (Los Angeles )» She 
research was directed prixaarily toward two goals? (1) the developssent 
of reliable stressor systeast for the production and raaintenancs of 
behavios-ally evident ©aofloaal dtsturbaac® and (2) the application of 
these systenjs, in an atteaipt to esmlore their effect on the growth of 
ti-ansplanted tuiSKsrso In a series of ei^t e^jerimenta, mice with 
subcutaneous Sirlieh tmsor traaeplatxts were subjected to stress either 
by an avoidance shuttlebox or by confineiseato In five of tiie eight 
esqperlasntss significantly ssBiller twssxcs were found in stressed 
aMaals as coan-ared to their coxitrolso T^o of the; reiaainlog tm-ee 
experiments yielded nonsignificant reai^-:^ in the sbsiq direction, and 



m 
% 

I 



- 13 - Annual Report 

kB-NCI 1959 

one showed a algnificant iner«ase in the size of the tumora,. Escposrire 
to ahuttlebox or confiisement stress tends to inhibit the grovth of 
Shrlich tusK>rs in siice, and it is sxiggested that stress represents a 
varial)le to be considered in cancer research* Stress also offers an 
additional context for the e:q)loration of tumor-host interrelationships » 

When "foreign" tissues {antisens) ere injected into sn aniutal, 
the latter responds by producing agents (fflatibodies) which atten5>t to 
destroy the intruder. Specific antibodies to particular tumor types 
have been demonstrated by several investigators o In studies on these 
ifflimmological reactions j, blood seruziss containing antibodies (antiseruffls) 
vere produced in White Rock and Hew Haispshire Red cockerels against 
the csaacer 391 mouse aaelmioffia and against tissues of noraal mice of the 
inbred tuasor-host strain (DBA/l), by Drs. Peter Abrenoff, Harry Chinchinlen 
and John W. Saunders, Jr^ of l^arquette University » Initial experiasents 
clearly shoved that the chicken responded iBammologically to injections 
of noM3sl and cancerous souse tissue prepai^tions in laich the seaae nsannor 
as it did to the injection of purified soluble-protein antigens o A single 
injection of antigen evoked a peak of antibody production in 7 to 9 days; 
a second iajecticei of hoisologoua antigen, given several weeks later 
called forth a peak aaatibody production at 5 days "ahich asay be ujaintained 
for as long &s 3 veeks« Pooled ssuoples of nortaal chicken serua, tested 
vith noxi^l and tumorous mouse-tissue antigen, did not respond by ex- 
hibiting a precipitate (precipitin reaction). On the other hand, both 
the aorsaal ^:d tuaorous tissues elicited appreciable, but low, precipitin 
titers in all birds (2^ '^^^ Saaroshire Reds; l6 White Rocks) tested, 
Iherc vas clear evidence of distinctive antigenic con^onents in both 
cancerous and norssal mouse tissixes; the S91 melanoma and normal tissues 
of the host laouse strain possess distinctive antigezrlc con^nentSo 

In their research on cancer, Drso Bill Pa Maduras, Steven Oo 
Sch»arts, and ^toIA H. Sdsoolss^i of the Hedtcxm Institute for )4edical 
Research have devised egperlmfmts to stuc^ the possible protective 
effects of cell-free brain filtrates a^ttast cancw, the filtrates having 
been irradiated «lth ultraviolet light. Hybrid mice (a cross of the 
C3H and 101 strains) from eight to ten tieeks old, vere -tused in the eatperl- 
aents. Brains of five hybrid CSSslOl nice vere pooled, and tram them 
1000 ce of cell-free brain filtrates was prepared. A control filtrate 
was siadlarly prepared with brains of nonleukesBic CSSslOl 35iic9o Ihe 
filtrates were irradiated vlth ultraviolet light. All escperliaental Mce 
were iuaaunized ^th ultraviolet irradiated cell-free brain filtrate by 
inoculations under the skin of 0.1, 0.25, said 0.5 cc at k6 hour intervals u 
5!he mice -were che^^iged 10 days after this last inoculation by abdominal 
cavity injection of 0.5 ce cell-free bi^dii filtrate or tuinor cell svis- 
pension prepared fs-cm leukeiaic CSHxiOl mice. In tersass of results, this 
active iOTamization 'fe'lth ultraviolet irradiated lerokeinic eell-frec brain 
filtrate gave significant protection a^lnst challenge of leukemic cell- 
free brain filtrate. Active isaauniaation with ultraviolet irradiated 
cell~free brain filtrated did not protect against challenge of a high 
ooneentratioa of tuiaor-cell suspaasioao 

290 



- l^t - Annual Report 

RGB-NCI 1959 

Co DetectloB, Diagnosis, and. Epidemiology 

Sarly detection end accux^te diagnosis and localization of 
tumors is, of course, important for adeqjiate therapyo If the 
presence of cancer can "be detected before it has invaded nearby 
tissues emd has spread to other parts of the body, its eradication 
by conventional sxirgery aaid radiation is more likely. The importance 
of epidiBsniological studies lies in the fact that they laay give leads 
as to the various causes of cancer and thus aid in its prevention. 
tost of the work in these fields, particularly in the more applied 
aspects, is supported through the Field Investigations and Demonstra- 
tions Branch, 0336 two projects described below illustrate sonse aspects 
of work in these areas that is supported through the Research Grants 
Broncho 

Dr» Paul Eo Steiner of the ISaiversity of Chicogo, in collabora- 
tion vith Dr., Robert CSeaain snd. J» Netik of the Inatitut Pasteur at 
E^er, French West Aftriea and Hopital le Dantec at the latter place, have 
investigated the occurresie© and the pathology of huaan liver cancer in 
Ifetkaro Dakar vas selected as one place for study because of the large 
amount of material available » These investigators have reported on 238 
cases of priaary cancer of the liver a The clinical course of the 
disease vas usually short and characterized by ebdoiainal pain and 
swelling, palpable hhss, jaundice, and digestive disturbwices in various 
coaibJLnations^ Dissaaination to other parts of the body vere recorded 
in only about half the cases, the lung being the most frequent site^ 
About ^^ of the typed ca9es were of the cancei-ous type designated as 
hepatocellular carcinoiaao Ruelear vacuoles appeared in some instances, 
and bile was seen in nearly tolf the tumors. Fat was evident in many 
tusjior cells; and there were on occasion, atypical cell fonaa. No u3Qiq.ue 
features vhtdti would separate these liver tumors from those occurring 
elsewhere, were encoisateredo The patients wejre all native Africans so 
far as known. ISieir average age was only thirty-six years, or about two 
decades younger than the American average. (Dto Steiner la now at the 
institute for Cancer Research in Philadelphia ) 

Some epidemiological features of leukeania in children in the 
Buffalo, New York, area have been studied by Dto Donald Pinkel of the 
Roswell Park Mamorial Institute and an associate o The total nunber of 
cases of childhood leukemia in the city of Buffialo for the period 
19^3-1956 was 95, with an additional U2 in the suburbs, making a total 
of 137 cases. 13iere were 70 boys and 6? girls in the group. Bie jo^xx 
age was 5.9 years. ISas incidence rate in children under I6 years in 
the city was 70 per 100,000 for this Ik year period. Sie mean eaim^ 
incidence rate wss gs^eatest in the highest intxms groig) (first quartlle) 
and lowest in the least econoMcally privileged g.uartileo Eiis tread 
is not considered signific^at, because of the smsuLl nuB&er of patients 
in the sample o However, childhood leukemia incidence in the aiffalo area 



29i 



- 15 - Annual Report 

RCffi-NCI 1959 

corresponds closely to figures reported in other epidemiological 
studies in the United States <> The higher incidence rate in the 
upper economie half of the population is ccaisistent vitdi results 
obtained in a study of leiakemla in children aod adults in Baltiaiore, 
Mo dose tearporal-st^tial relationship of residences of children 
with leukeaia was noted, eaid the frequency was suggestive of a 
cojmminicable infectivity. 

Do Treatnssnt 

Che«K)there!^ of cancer continues to attract increasing atten- 
tion and an increasing proportion of esvailable supports Ihe use 
of honaones plays an important part in the treatiosnt of several pre- 
valent types of caaccTo !Ihe application of iiaBunological techniques 
to the treatesnt of cancer showo a great deal of proaise but it is 
still in the laboxBtory stage « Despite those lEore recent develop- 
ments, surgery and radiation stiU. are the only ffisaas of cxarlng 
cancer aod iSiese nsethoda are effective only when the disease is 
iocalizedo More work is being done in a relatively new approach to 
cancer thejrapy, the use of transplanted autologous or hoiaologoue 
bone laarrosr, 5!his technique peirolts the use of larger, and otherwise 
lethal, doeses of drugs or radiations » After treatfficnt is canpleted, 
the patient or animal is re- inoculated vith tint rested bone icarrow 
previously obtained f^om the seaae patient or anisal or obtained fsxaa 
a closely related source. 

It is su^ested by Drso William MciPariand, ISoras, Bo Grssaville, 
aind Williajs Daajeshek of the Key England Center Hospital end the !i?ufta 
Universii^ School of ^5edieine that att^aipts to ei^dicate nslignaat 
tissue by nonsurgical method (X-rsdiatlon, cheBotherapy) are frequently 
liBJited by the vulnerability of normal tissues, particularly bcaie sarrow, 
gastrointestinal tract, and skin« Such considerations are partictQ.arly 
pertinent with respect to lym^ihos&iccixaa, and Hodgkin's disease » To date, 
folloving nssaive X-ray or cdiemothei-ai^ in hutaan beings, efforts to 
repopulate narrow by saeans of bone Bsarrcfw transplantation from other 
individuals havtf been uniformly unsuccessful, probably because of the 
develofpaeat of an iazaune reaction betveea host and transp2,ant. !l5ie 
\tse of the patient's (autologous ) bone sasarrow in selected cases circum- 
vents this res-tion and at least theoretically should permit higher 
dos©^e of X-iradiation or "chezcotherapy with's^iwisequent bone marrov re- 
covery. They report that three patients vith advanced lymphosarcoaas 
and two with Hodgkin's disease ware treated t.Jlth massive doses (super- 
dosage) of nitrogen mustard (over 1.0 milligram per kilogrsm) given by 
vein, folloved by infusion by vein of autologous bone laarrow^ T\-ro of 
the five patients succuiabed to infection. Bie resaaining three patients 
recovered within one month and experienced brief remissions <> 



292 



"16- Aonual Report 

ROB-NCI 1959 

It is suggested tlmt this fonn of therapy merits further trial. Thus, 
it njay he considered for use earlier in the course of the disease when 
there is a saaller total aiaount of tumor tissue, and thus a greater 
possibility for its complete eradication. Its use in other types of 
tumor disorder, ae follosrtng operative procedures disclosing malignancy, 
is suggested. 

MsaiBBals of several species can be protected against otherwise 
lethal doses of whole-body X-irradiation by post-irradiation adaiini- 
stration of bone marrow. Protection is derived from the repopulatlon of 
the blood-forming system in the bone marrow by cells introduced from a 
naarrow donor, regardless of whether the protection is achieved with 
isologous (sains gene patterns), hoinsologous (same species), or heterologous 
(different speeiea) bone nsarrow. Bone marrow from the BALB/C and CBA 
inbred strains of mice and the (BALB/C x CBA) first generation hybrid 
strain vas injected by Dr.. J. J« Trentia of the Baylor University College 
of Medicine isologously and in various hoinologoui combinations into CBA 
and (BMJB/C x CBA) first generation hybrid mice irradiated with 330 
roentgens^ 550 roentgens, or 770 roentgens. At the sublethal dose of 
330 roentgens no saortetlity resulted from any combination of laarrow donor 
and recipients At the higher dosages, two types of mortality were 
observed in mice receiving bone raarrow; (a) "early mortality" (5 to 21 
days) after the oi'dinarily high sublethal dose of 550 roentgens and (b) 
"delayed ssortality" (21 to 60 days) following good, early protection 
against the othervis© lethal dose of 770 roentgens. Heither type of 
laortality occvwred after administration of isologous marrow, but both 
types occurred after administration of marros' from an unrelated hcMnolo- 
gous strain. When raarrow was transferred frcm first generation hybrids 
into the parental strain, only "early taortality" was observedo When 
parental marrow transferred into the first generation hybrid, only 
"delayed mortality" was observed.. From these observations and other 
considerations it is concluded that the early mortality, resulting 
from administration of homologous or heterologous marrow into mice 
irradiated in the high sublethal or midlethal dose range, is the result 
of the recovered ability of the host to reject the temporarily grafted 
nfflrrow, whereas the delayed Jioortality, following good, early protection 
against otherwise lethal doses of irradiation with homologous or 
heterologous narrow, usually results from an inammologic reaction of 
the grafted marrow derivatives against the tolerant host. 

Drso E. Etonnall Thomas, Harry L. Loehte, Jr., and Joseph W, 
Iferrebee of the Mary Imogene Bassett Hospital (Cooperstown) and the 
Children's Cancer Research Foundation (Boston) note the infusion of 
bone marrow will induce recovery in several animal species after 
exposure to irradiation that otherwise would prove lethal. The 
question naturally arises Aether nsarrow spaces and soft tissues in- 
fested with leukemic cells can be siJsilarly cleaned out by diffuse ion- 
ising radiation and repopulated with normal cells » With some strains 
of leukemia in some strains of mice, this has been accos^lished. 

293 



- 17 - Annual Report 

RGB-HCI 1959 

^ese researcbers nov present clinical eacperl«nce vith vhole-body 
irradiation and loarrov transplcmtstion in mano Tests vere reported 
from seven cases ^ including four raeai froaa 6l to 2^ years of age and 
three girls aged 3, k, asad. l6o Poia* patients (one laale, three female) 
had an acute leuk^ao One nale had a layeldblastic type of tui&or 
(in^ture heme marrow cells }j one a chronic lyiHgiiatic leukemia, and 
one a cancer vhicb had disseminated to the lungs; this last patient 
vas not irradiatedo Ihe remaining patients, most of vhom had had 
relapses after chessotherapy, ^'ere irradiated vith tissue doses ranging 
from 250 to 6(X> roentgens o This was followed by intravenous infusion 
of hone norros', either from close relatives or frOTa individuals who 
had Just diedo It vas difficult to establish complete genetic ccm- 
X>atibllity, and in at least tffo cases, the transplantation of narrov 
cells Has not successful > ^ere the test itself vas successful, the 
Idulcemic mssraw vas to all intents and purposes destroyed by the 
irradiationc Tr^oefusioBs of vhole blood vsre necessary also isaoedi- 
ately after irradiation, to tide over the period of naxrav redevelop- 
Bsste In the one isale patient not irradiated, nitrogen mustard vas 
used in lieu of irradiation, and he vaa then given oarrcw cells; but 
infecticm Bxxs^rveasd said he died of an overuhelzBing infecti<xio 
Siajilar infection killed four of the other patients because of the 
exbre!iSS and unrelie^^d reduction in vhite blood cells caused by the 
irradiationo In two i^tients, one feisale aged I6, and one male aged 
59* the treatment ve^ regarded as initially successful, but t2ie 
inale patient ev^itually ouccuiabed also to infection. 

Bse three projects described below illustrate the kind of work 
tha,t is being dona on the use of bonsonss la the treatment of cancer <> 
The response of cfiaicer of the prostate gland to hormonal therapy has 
been termed probably the most outstanding jdienooB^on in the study of 
cancer. Although it is coBBacmly recognized that hormonal therapy such 
as castration, administration of femsLLe sex hormones or removal of 
the adrenal gland tends to alter the natural course of this disease, 
the chaagea ^lich occur upon therapy aare only partly uaderstoodo DrSc 
Vincent Jo ©•(tonor, Jr., Robert E, Desautels, June W, Pryor, Paul L. 
Munson, P^a.P, ^id J. Hartwell Harrison have recently reported on 
studies of honaonal changes' in relation to cancer of the prostate 
undertaken to develop knowledge of the applicable -endocrinologsr. 
Patients xinder care and personnel at the Peter Bent BrigheuB Hospital 
in Boston vere subjects . I^5ale sex hormones (iT-ketoateroids and 
androgens) ahoiv a definite decrease with advancing age, both in noraal 
suibjects and in patients, and there is no essential abnormality of 
androgen productic«i in the prostate cancer |«,tlento Androgen 
decreased consistently by about 33?^ after castration in every case 
(12 {^3eo)o The regain ?.»g androgen is believed due to the adrenal 
gland. Upc» stimLating adrenal foncticn vith the adrenocortico« 
trophic hormone derived from the pituitary gland, the amount of 
andr£@sa will Jacireaseo After adrenaleGtoagr, when large doses of the 
adrenal horEone cortisone are given, androgen excretion rises in 



29U 



- 18 - Annual Report 

ROB-NCI 1959 

aliaost a step^se fashiooo Androgen excretion is significantly 
rediiced in psti^its receiving female sex hormones, both before 
and after castration. Continuation of ccniasined castration- feoale 
sex horjQone therapy is suggested as the initial ideal honacxial 
treatxaent for csaicer of the prostate., 

MsLle sex bowQones have been effectively enpli^ed in the treat- 
ment of advanced breast caaacero Dr. B. J. Kennedy of the University 
of Minnesota Msdical School vrites that objective iniprov«3«nt occurs 
in approxiBa4;ely 25?* of patients vith dissemination of cancer to bone, 
with a lower rate of response in patiaats with soft- tissue lesicajSo 
Be treated 56 woaen vith dissezoinated breast cancer over a two year 
period with a honiKmal derivative^ fluoa^assterone^ Of these, k6 
had received no previous form of honacme therapy « The results of 
therapy vith this coa^pound shoued -Uiat fluoxyraestercme is an effec- 
tive agent in the tr^itment of advanced breast cancer. Since it is 
an oral preparaticsi, it is easier to administer than Intraimiscular 
prepaxtitic»js. 13ie emtitvooor effect Is at least as effective as that of 
testostercHie, propionate ^ and trcesi these preliiainazy data in a soall 
nuzzS^er of patients, flua^aesterone say be superior to testosterone 
propionate. Cfoservaticms of the side effects of fluoa^aasterone 
reveal nissculinizing changes, though the intensity of these side 
effects is not as great as with testosterone proionatea Fluoxy- 
neaterone is an effective androgenic hormone In the treatment of ad- 
vsmced breast cancer snd nay be considered the male honoonal 
derivative of choice at pres«at« 

The r^fioval of the pltultajcy gland has been evaluated for its 
effect on disseminated msmmary cancer by I>rsc Olof Bo Pearson and 
Bronson S. Bay of the 31oaa-Kettering Ii^titute for Cancer Research » 
Tso hundred eighteen patients vltli disseminated fflsnoary cazKser hsd 
bypophysectcsaies betve^i I<jsrch, 195^^, and >^axh, 19580 Ibe results 
for 109 patients vho had a minijcaum follov-i^ period of 1? xsoaths 
are rejportedo Approsdjaately 50?^ of these patients obtained objec- 
tive ranissions, s^kI 35^ had remisslooa that have lasted 6 aioQths 
or longero In the latter group, the average period of r^aission was 
15 plus laonths end the average survival tizae vaa 21 plus saonths. 
A prior fBvor«*le response to siurglcai reiaoval of the ovaries was 
found to be of prognostic value in predicting a beneficial effect 
from hypopl^s€Ctany» In this setting, hypophysectony yielded better 
results than did edrenalecton^o She adioinistration of f^&ale sex 
horaooe to 5 patients after hypophysectooy f)alled to Indixee 
exacerbation of tunsor growth, suggesting that a factor froa, or 
mediated by, the pituitary is necessazy for female sex honamie stlmu- 
lation of tumor g3:'cnrtho Huzoan gra:?th horaxme administer^ to 5 
patients after hypos^sectomy appeared to induce stiiuuiation of the 
tun»r growth la 2 i«tientSo These preliminary observations suggest 
that groifth hormone nay be an in^^rtant endocrine factor in ESMSsary 
cancer %pophysectocv is reeoEBisnded as a practical, worthwhile 
proeedurs In the palliative treatirient of patients ^=d.th disseTniaated 

295 



- 19 - Annual Report 

BSB-SCl 1959 

breast cancer ^en adequate facilities are available to carry out 
this procedui'eo 

One of the great die&ppolnticeQts in the treatment of oancer 
has been the failure of chenotherapeutic agents to cure or even to 
produce sustained palliation of saoet ssaligpiBnt lesions <> ^Siis ie due 
in psart to the fact that availelble drugs are toxic to rapidly grow- 
ing normal tissues viisn. administered in eitfective aaiountSo Within 
the past t»o yeejrs B . Oacar ^c^®ch> Jr» , and his colleagues of the 
Tulaae I&iiversity School of Medicine have developed methods irtiich 
psnait the adBsinistration of larger amounts of cancer-destroying 
dsmgs and at the ease tisie reduce the danger of systemic toxic 
effects. trSaese saethods consist of techniques for isolating and per» 
fosicg the liaibs, l\mg, pelvis, and breast in the treattoent of 
regionally confined eaaeero A method of perfUalog the entire body 
for treatsisnt of disseminated tuusors has been developed alsoo During 
the ^^ast one and one^half years 73 patients vlth noalignant tumors 
hm'e been treated ^th ehemotherapeutic ag^xts administered by these 
teclmiquesb 'Bie agents eo^lc^ed Offers nitrogen mustard, phenylalanine 
atustsrd, triethyleaethiophosphorandde, actincHo^cln-D, and 5-fluorour- 
acilo Fifty-six of these seventy- three patlexits vere treated for 
palliation of fsT'^advanced zaaHgntsncies and sevente^i vere perfused 
in coajuaction vith extirpation of the pi*iaffitry lesions <, TMrteen 
pati^ats have died; aiaong the sl2£t^ patients reo^ining, tventy^tw© 
are well snd the lesion appears to be controlled; in eight the 
disease is quiescent and in thirty it is recurrent » 

Has coBsbination of chemical agents vith radiation therapy has 
often been tried in order to attain inci-easelttntitxflaor effects, and 
increased Inhlbiticai of cancerous processes has beesi notedo In a 
recent resear<^ project, Ers<, Daniel 3. Martin, Msrtcai M, KltgenMua 
and Buth Ac Fugmosm of the Coluit&ia University College of Fhysidans 
and Surgecms have been able to densnstrate slMlarly augQient<sS 
r-aaissicas rate® by coatoining two clinically acceptable ccmtpouods, 
6->iaercaptcpurine and S-aalncMjicotiBMdde x»ith i^dio thers]CQr<. Male 
C^BXi £2ice, S to ^ jssonths old and 'neighing l8 to S3 grasB, vere zoain-* 
tained under optiiatffii cooditiocso ViBsmsxy adenocarcinoma 7^5 ^ijaa 
tranaplaated into the subcutaneous tissue of i^a& left hind leg of 
each animal. ISiese l^laats were alloired to grow for 17 de^s, at 
i^di tlBie t^ey vere veil established end of good size. Beginning 
on the 17th day after tumor trsaasplsuatation, all aalinals receiving 
chemotherapy, alone or in conjunction with radiation therapy were 
treated once daily for 5 consecutive days; 6-ffiercaptopurine and 
6-amiaoaicotiaQJBide were injected into tdie bodly cavity in carefully 
calibrated dosages.. Following the five-day treatment, the cminala 
-wBve observed for from 33 to 38 d^B, then sacrificed, and all aoa- 
regressed tussore earefaily wei^edo It was found that mlaiiEal 
chemotherapy acted as a powerful adjuvant to radlotheraKr in produc- 
ing tuiBor dssija^o Sis average percentage of tumors "cui-ed" was 
5iicreased frons 27 per cent trlth x-ray alcme, to 63 per eeaat with 

29G 



- 20 - Annual Report 

RGB-Kd 1959 

6-iiiercapti^urine plus 6-aEsiaonieotinamide pliis x-r^o Taese 
researchers feel that the documented results recoaunend similBr 
methodology I'or clinical evaluation of certain potential cancer 
cheisotherapeutic agents. 

The following report is from the Acuta Leukeaia Cooperative 
caieEKstheraapy 03?oup B of ^ich Dr. Eniil Frel of the Haticmal Cancer 
Institute is Ghaiirasaxo A coaiparative clinical trial of two regiaafins 
of codbinatitm chemothera^ has "been accocrplidhed in treating acute 
leukeraiai Sixty-five patients were allocated at random to one of 
tisto treatisent progri^as. Daily adMnistration of methotrexate vith 
daily 6-xissrcsptopurine has "been ccsEpared to aethotrexate every third 
day in the saaae total dose vith daily 6-ffiercaptopurineo So differ- 
ence in freq.uency of remission, eartieat of remission or toacicity was 
observed between the two groups o AjEong those patients who attained 
remission status ^ however, duration of r^nisaion and of survival Ha@ 
longer for the continuous groupo All remissions in cidldr«i occurred 
in acute lyii^ocytic leukestia, t&ereas all ranissiots in adults vere 
observed in acute isgrelocytic leukesaia,. 2he duration of remission 
Wis Sdrsewhat shorter for children ulth acute lyiBpho<^io leukemia 
than for adajlts vith acute n^relocytic leukesaia. She firequency of 
realssion, either partial or conrolete, was higher in children, 
however, (3^), thsaa In adults (195»)* although the confidence 
lijaits for each figure overlap. !I!he duration of acute leukania in 
previously untreated patients did not influence response" to therapy 
frcjsB the t^yo methods used in this study,, In patients ^ftio previously 
had had theragy of the e^a» sort, howsv«r, cosaplete reroissiona vers 
attained less often then in previously unti-eated patients. 

^e t(»d.c zi^mifestationa encountered during the adzainistra» 
tion of these antistetebolites are described. Seventeen deaths 
occurred during this stiady, of "ahich eight occurred in the first 
ten days, presunsBbly from leukoma and not drug toxicity. She toscic 
namifestations ^ere qualitatively ©nd proportionately the sasne in 
patients vho attained remission status, end in those patients vho 
flailed to reait, but \Jho lived long enou^ to recognisse the onset 
of remission if it were going to occm*. Ko indlcatitm was obtained, 
therefore, that patients who attained remissiisi vere subject to a 
greater toxic hazard, in order to achieve the therapeutic benefits 
observed, than those who did not remit. Hie aediaa survival of 
patients Tiho achieved raioission was longer thao for patients who 
did not rsatto Since the survival time of x-eaittei^ from relapse 
to death v&b alnjost identical vith the survival time of nonreaiitters 
froa <xiset of treatss^t to death, this difference can "be accounted 
for by the time spent in r&oisslon and getting to x^^sslcaa. 1!he 
median survival time f3rom syis^stoiEatic onset for all children in this 
study vas twelve toonths, and for adults, seven UKjnths. Bae isedian 
in children is siiailar to that reported from other clinics « Tbla is 
evidence that a ccarp^rative therapeutic trial in acute leukaais csa 
be aecoEplished vflthout recognisable cosaprOTsls© of patient •<relfsreo 

2S7 



- 23. - Aanual Report 

R(3B-ECI 1959 

A nxnnber of ccsapouncla, aiaong them cysteine, cysteamine aad 
sodixm nitrite, afford significant protection in experimental 
eminsBls against a lethal dose of x-rayo, especially when admini- 
stered prior to ir2'«diation» ®ie nitrogen naistards CHH2) caad 
related ccanpounds su<^ as triethylene melBBJine (ISEJM) show cancer- 
inhibiting and pharmacological actions aiiailar to those produced 
by x-rays aad have therefore been termed radioorfjaetico'' Drso 
Edsd-n Xo Golden-ihal, Mo V« Sadkumi, aad Paul Ko SBdth of the 
George Washington Uhiversity School of Medicine have made a compari- 
scm of the protective action of cysteine, cysteaialne, sod sodium 
nitrite esainst the lethality in mice by x-irradlatioai, nitirogen 
Bsustard, or triethylene laeilajEdneo Striking differences vere 
revealed in the degree of protection obtained by the preadministra- 
tion of the coHpoundSo All eontpounds afforded some protection against 
the lethal effects of x-lrradiation. Si&gtaatial protection against 
the lethality of ESi2 was obtained with cysteine, and some protection 
with cyateajainec Hone of the cca^pounde gave oeasurable protection 
a^lnst lg®i. It was found possible to initiate the chemical inter~ 
action of ®2 with cysteine or cysteamine in tissue ct2ltur«o ®ie 
reaction of M2 with cysteine or cysteaajine was instcaitmaaouss, 
whereas that bettfeen 1SS6. aad cysteine or cyst^anine required several 
hours It is si^gested that the difference in reactivity xnay ejqplain 
the dlffesrences in the degree of protection afforded by the sulfhydrul- 
containing cc^ipoundso (She tissue culture reaction product of OSSM 
with <^steine was isolated and pua^lf iedo ^Sils ccmrpound was shotm to 
be inert with respect to toxicity as wall as cancer-inhibiting 
effects on leukemla-12iO in micso A threefold increase in the urinary 
excretion occurred wh^i cysteine was preadainistcred to loice as casa- 
pared to BH2 alone « The suggestion is sade that the isechanisia of 
cysteine protecticm is through direct chendeal InactivaticMas of HB2o 

la the search for therapeutic drugs trtdch may be effective 
©gainst various types of cancer, Dr» Erich Birschberg of the College 
of Phyoiclaos ^sd Sxsrgeons of Cblunft>ia Ibiiversity and his colleagues 
have investigated the effects of mlracil D, amodiagLuin, and 22 
chemieall^ related ca^oundSo Miracil D, a member of a chesaical 
groizp kno«n as 10-thlaxanthenones; et^iibits a wide range of c^acer- 
inhibiting effecttvajeas against a variety of treasplantable mouse 
tumors at nontoxic dc^age levels a Sarcotaa l80 and the lyoiphoid L 
1210 ascites leukeaia respcsided most read? 1^, followed by adeaocarcinoBnas 
755 and 230771o Qlioiaa 26 aad the Ehrlich ascites carcinosaa EBnifested 
a narginal response at best* Adndnistration by mouth appeared to be 
preferable la scses instaaees to infection into the abdominal cavity., 
Affiodiaquin had only borderline cancer-inhibitory activity in the s^oe 
tumor spectrumc She only tmor found to res^pond signiflceatly after 
oral adujiaistratioa of this agent was adeaocai'ciacsis 755" K^e o^ier 
10-thiax^ith®aones aad U-emir^guinollnes have been compared to these 
two coa^wunSs, in teraas of relative cell toxicity in tissue cultures 
of hiasffia sad saouse brain tisaors, maximum- tolerated dose levels ia 
the mouse ^ aad cancer- inhibitory activity sgainst a variety of mouse 
txjirorso Son© of l^e activity of Miracil D in the living body, and 

298 



- 22 - Aimua]. Report 

ROB-NCI 1959 

none of the ajoodtaquin analogues ^ms superior to anK>diaq,uln in thie 
regardo In tissue culttire, njany of the 10-thiaxanthenones vers 
more active than the parent congpoundi there is no correlation 
between activity in the aaiiaal ami in tissue c\iltiare» 

Over 130-cheBJical coK^iounda have been tested for possible 
cancer- inhibiting activity by Di-So Arthur Furst, Windsor Cutting 
and EJidi S., Gross of the Stanford University School of Medici ae, 
Swiss -Webster Mce of either sex were inoculated in the abdcaninEa 
cavity with the free-cell ascites cancer taiai fron a 7-day tusKXTo 
Agents vhieh, «hea adiainistered to tumor-bearing mice, increased 
longevity by al; least three days from & contiol period of about 
11-13 days 5 were considered to have significant inhibitory action 
upon this souse tuasoro ^ey included pyrrolidcmej, 5-.nitrc-2- 
furfuiuldiacetate, vanillal E^sityl oxide, S-fUroyl furylasaiae^ 
C5-hy(3raxyj2athylfuryl} acrylic acid, 2-JBethyl-3 Icetosuccin dismide, 
jaalcnonltrile, S-nitro-J'^s^csiiodiphenylaajine, paraethoxyazobeaizene 
aod para-chXorophenyl^clohexyl earbinolo Maxinsum extension of 
longevity (6 days) occurred with 5"nltro-2-fUrfuraldiacetate mvd 
vanillal mesitylosideo 

Budget Data aa Research Grants 

During the calendar year 1959* the !iatlc«al Advisory Cancer 
CtouuGil revievred applications aaad recoansendad researdi gMttits as 
follows: 

Htadjer Amount 

Applications reviffirad 2,25^ $^,709,015 

(k-fisits reccsOTended 1»900 36,136,630 

Siese figures include 1,128 grants for $^,853,308 which had been 
previously recoiaascded for moral ccsaaitaent of futtire svgjport in 
fiscal year l^Oo Shese recoesjjendatlons were reaffinaed by the 
Council at its June 1959 saeetingo 



299 



- 23 •- Annual Report 

RGB«NGI 1959 

Raseareh FellCTTshlps 

Ths Rosaarch FallotEvShips program of the National Cancer Iinstitute 
supports the raseareh training of wsll-qualified csndidates in basic 
science and clinical fields partinent to cariear £3o that mora and bettor 
qualified inx'sstigators will be e:fiList©d in the attack on tho problems 
of cancer at all levels. It parallels similsr pi'ograiiis in other Insti~ 
tutes and DiviaioaSj but differs in the fields in which awards are 
made. The program finances predoctoral, postdoctoral and special 
fellowships and part-time medical stndeait f ello>?ahips . It provides 
sn)a3.1 ^BSits to daf$"ay th® costs of supplies and similar educational 
expenses 

Predoctoral Fellows already tiave made theJx plans for careers 
in ressareh* Thoy are embarked on a progrgjn of braining in research 
methods and conduct of research that will lead to an advanced degi^ea 
or its equivalant. 

Postdoctoral aild Special FoXlowa are. expaiiding' their rsseerch 
tra-ijiing and Q^qjerience under the supervision of or in association 
with an established scientist. This prepares IJ-sem to take their 
places as investigators axid teachers in the research and educational 
institutions of the eoimfcry. The currently recognized shortage of 
adequately trained research personnel to staff these institutions 
€!nphasizes the need fos* expanding and constantly replenishing the 
pool of researoh manpcrwer on which progress in madical and scientific 
research depends « Approxijnately 90^:^ of the scientistfi previously 
trained under this program are now aetivoly engaged in research or 
teaching, or both» of raedical or biological subjects related to 
cancer. 

The part-tirsQ aaadical student fellowships ars part of sm iivber- 
Institute coordinated program designed to give ixidividuals interested 
in a resesrch career the oppar-timity of aarly participation in estab=> 
lishsd research programis during the sunsaar or other pariods of the 
year when they can interrupt their forraal raedical training. It is 
hoped this experianes will stimulate their contiaiuing interest and 
devalopRssnt in a research career. 

The following table summariKsa feLloi«rships5 awarded in calendar 
year 19:59: 



300 



" 2k " Annual Report 

RGB-NCI 19^9 

S^ Number Amount 

Full-Tisne: 

I^edoctoral 190 $ 559,U3li 

Postdoctoral 155 977,717 

Special _^ 139,225 

Total, full-tiai© fellowships 363 $1,676,376 

Part-time medical stiidents 256 165,888 

Supply grants & special allowances 6I 29,375 

Total Awards $1,871,639 

Thus over 6OO scientists or predoctoral science students were assisted 
in obtaining . research training in basic and clinical sciences bearing 
on the cancer problem, including many aspects of biology, chemistry, 
physics, and medicine <> 

Research Training Program 

Objectives of the research training grant program of the 
National Cancer Listitute are to increase scientific manpower in clini- 
cal, biological, ehsmlosl, and physical disciplines and areas of 
investigation pertinent to research on the problem of cancer, its 
cause(s), diagnosis, iiierapy, and prevention. Manpower shortage in 
many areas and disciplines important fop cancer research, botii at 
present and in the foreseeable future, makes it imperative to train 
research workers who can increase the research potential as well as 
replac© present in^restigators who are being lost to cancer research 
through dea'tei and retirement. 

The program began in calendar year 1956. Hith enthusiastic 
and unstinting assistance and advice from current and former members 
of the National Advisory Cancer Council, the program has been developed 
to meet present day needs. Although there is some evidence of 
immediate greater need in son® disciplines than in others, the 
National Advisory Cancer Council Committee on Research Training 
believes that the rraads ere so widespread in nearly all pertinent 
fields that ncaie should be excluded as a potential area for training 
of scientists \riio will contribute to new knowledge regardirig cancer 
and its control. Specifically, however, special interest is directed 
toward increasing the number of well-trained inrostigators in clinical 
research on eaaeer chemotherapy and pathoplgrsiology, steroid biology 
and biochemistry, biophysics, experijaental chemotherapy, pharmacology, 
genetics, analytical cytology (including electron microscopy and 



301 



- 25 - Annual Report 

RGB-NCI 1959 

histoGheiaistry) , vixTises and immunology, radlobiology and radio- 
therapy. Ot&es' areas of ik) less importance include biochemistry, 
biology, euq^erimental pathology, sistabolisra and nutrition, and 
experimental surgery. Training may be wholly within a discipline, 
interdisciplinary, interdepartmental, or interinstitutlonal. 

In general, the program contemplates training at all appropri- 
ate levels that opportunities afford. These nssy range from advanced 
special training through postdoctoral and predoctoral stages to 
special programs for training of technical personnel and of teachers 
at the college and secondary school level, and for developing the 
research interests and potential of medical, premedical, prebachelor 
and preeollegiate students. In view of the limited funds available 
to roB&t the requests, the Council has for the present given more 
favorable consideration to -Wfcxose proposals that would provide train- 
ing at predoetoaral, postdoctoral and advanced levels, 

Reseaareh training grants allow Uie director of a training 
center to select the trainees and to set the trainee stipends in lino 
with the pol3eyof his imtitution. In addition to stipends, the 
training grant inay provide for additional staff, research equipment 
and supplies, and other costs essential to the training of research 
personnel. 

The research training program is complementary to, and does 
not replace, the mechanism of training research perscranel through 
direct research fellowships described in a preceding section of this 
report. It offers a means to ^cpand the facilities and staff for 
training* It introiuees, also, an element of flexibility to meet 
the variations in -fee needs of training centers and to facilitate 
the selection of research tz^ineesk 

The p2?ograin has provided much needed 8J»i soa^t after oppor- 
tunities for stinsulating and attracting students at all stages of 
development to explore nevr fields, learn new techniques and expand 
their horizox^ of kno^^ledge and interest. In many instances, train- 
ing has been given to students who have received no direct financial 
assistance from the grant. 

latere is a great variety of mechanisms by which research 
twining can be successfully offered, l^l&n^ of the trainees have 
participated in special courses in steroid biochemistry and endo- 
crinology, in genetics, in histochemistry, and in oliier fields. 
Such courses range in time from a few weeks to 6 months or a year. 
Other trainees have participated in research projects as ^vxilar 
collaborators with senior iavestigators. Still others have engaged 
in more or less independent research on their a»n problems but under 
the guidance of a senior investigator. 



302 



^ 26 « Axumal Report 

RGB-NCI 19^9 

The general and special fields in which trainees are engaged 
cover a wide spectrum of biological, biophyTsical and biochemical 
Icnowledge. The distribution of trainees in these fields has not yet 
been tabulated and ajialyaad* It reay be of interest, however, to list 
the fields* They include biochemistry (with special emphasis on 
enzymolo^, steroid biochemistcy, etc.), experimental surgery and 
diagnosis of cancer, Qxperiinental paidiology of neoplasia, histo- 
chemistry, cytology and cell physiology (using biophysical methods), 
biophysics with special reference to radiation, carcinogenesis, 
irsEMnology, viruses, experimental chemotherapy, clinical cheraotherajy 
and pathopiigRSiolos^ of cancer, pharmacology with reference to ehemo- 
thersjy, synthetic organic chemistry, nuclear medicine, endocrinology, 
and many more. 

Summer offers a rich opportunity for predoctoral graduate 
students, medical students, high school and college teachers, nurses, 
college students, and even precollege students to use productively 
s<»ie free time to explore their interests and expand their knowledge 
in a research environment x-rhere they are welccsaed. In a considerable 
number ®f training centers students from one or more of these 
varied stages of predoctoral research developnant were welcaned and 
took advantage of the opportxinities . It seems quite like3y that 
from such experiences maiy will be led into research careers in fields 
■Uiat provide knowledge toward a better understanding and control of 
cancer. 

Thus far, most of the training grant funds have been used for 
stipends to enable postdoctoral and predoctoral students to pursue 
their specialized training and their research problems. Some stipends 
have m^e it possible for medical students, high school and college 
teachers, nurses and college students to use their free time for 
research training instead of seeking gainful emplcytaent. Stipends 
have ranges from $100 or less for living expenses for a few weeks to 
$9,500 for a clinical investigator ^qsarienced in pathophysiology 
to spend a year lesmin^ about the probler?ffl of clinical investiga- 
tion in cancer patients so he can apply his rich knowledge of 
special physiolo^ to them. In addition to stipends, the grants 
have provided supplies, equipment and facilities to meet the needs 
of this greatly expanded program. 

Training grants provide a flexibility which allows the progi*am 
directors to adapt their programs to opportunities and needs as they 
become apparent. This feature has been largely responsible for the 
remarkable speed with which so maty trainees have begun their training. 

Prois cancer investigators who are aware of or have partici- 
pated in these programs, from program directors and research or 
educational administrators, tram trainees and from lay observers, 
liave come tmsolicited enthusiastic statements of the value of these 
training progr'ams in their initial phases. Most of these statements 
stigssst that tl'iera is a f^r g^ouitSj? ia-berest in scientiric i-esearch 

303 



- 27 -» Annual Report 

RGB-HCI 1959 

among the gesaeral population of students ia medical sdiools, colleges 
and aecondaz^' schools than is ccanaoiily realized ard that the oppor- 
tunities to deT©lop tiie potential of scientific manpower wore never 

greater. 

The follc^ing tabls suisssarizas applications re-^-iewed and 
recommended in calendar yesr 1959 t 

SItnaber Asiosmt 

Rsviewsd 65 $3,550,127 

Reeommendisd 57 3,1141,1*81 

Of tha 5? grants, 32, totalling $l,8lOJ^U53, were prerioasly reccsomended 
support prior to 1959 and wsrfe rseonf insod at th© Jtm© 1959 meeting 
of the National kdvlaoxry Cancer Cknancil.* 



30^ 



Serial Noo NCl-15-000 (c) 

lo NCI 

2o OABR 

3o Be^hesda 

PHS-NIH 

liadividual Project Report 

Calendar Yeas 1959 

Fart A 

Project Title: Screeaiisg 

Principal Investigator: Sr. Charles G« Zubrod 

Ofelier Investigators: Physicians participating in patient care 

Cooperating Units: General Medicine Branch, Surgery Branch, 
Endocrinology Branch, and Radiation Branch 

Man Years: Patient Days; 

Total: 

Professsional: 
Other: 

Project Bescription: This is a mechanism by which this necessary 
hospital function can be ciassif ied« 



Part 3 included Yes / / Ho /k / 



. 305 



Serial No<. NCI-15-001 (c) 

lo KCI 
2« CABS 
Jo Bethesda 
PK3-NIH 
Individual Projaet Report 
Calendar Year 1959 

ixt A 

Project Title: Service for other Insuitutes 

Fsiacipal Investigator: Dr. Charles Go Zubrod 

Ottisr iKVSStigators: Physicians participating in patient care 

Cooperating Units: Geaerel Medicine Branch, Surgery Brainch, 
Endocrinology Branch, and Radiation Branch 

Man Years: Patient Says: 

Total: 

Professional: 

Other: 

Project Description; This is a uaechaaism by which this necessary 
hospital function can be classified. 



Part B included Yes / / No /x / 



, 306 



■ . Bethesda 
PHS-HiH 
Individual Project Report 
Caleadasr Year 1959 

Fast A 

Project Title: Emergency Admissions 

Principal Investisator: BTo Chayles Go Zubrod 

Otter iHvestigators: Physicians participating in patient care 

Cooperatissg Units: General Kediciae Branch, Sargary Branch, 
Endocrinology Branchg and Radiation Branch 

Man Years: Patient ©ays: 

Total: 
Professional: 

Other; 

Project Description: This is a mechanisia by v?hich this 
necessary hospital fuacfcion can be classified. 



Part B included Yes / / No /x / 



307 



Ssrial Ko. £01 
Of f ics of Associate i/;.:. 
ia Charge of Research 
^tioBsl Cancer lastitute 



PHS-HIH 
Individual Project Report 
Calendar Year 1959 



Part A- 

Project Title; StDdies of Cytochemical Changes in Living Ii^ividual Cells 

Principal Investigator; George Z. Williaas 

Cooperating Units s Clinical Pathology DepartEsnt, Clinical Center 

Field Investigati^is wsA Qei{%sn0tratl<»:iS Branch 
Hational Cancer Institute, Serial Ho, ikk 

Departasnt of Chemistry, University of Richsaond 
Dr. J. Stantcsm Pierce, HCI Grant So, C-2693 

Man Years Patient Days; 

Totel s 3-25 

Professional; 1.25 

Other : 2 

Project Description; 

A„ OBJECTIVES 

The stisiy of changes of metabolism end chemical states in living IMividiisl 
cells by cytochemical methods is our sain objective. Ultraviolet Mcroscopy 
constitutes the chief instrument for analysis. The iisage is detected and 
transmitted by TV to a suxiitor for direct observation, and the intracellular 
reactions and structures and their ultraviolet absorption characteristics sre 
depicted and measured on a line -selecting oscillcscope . Burisig the past caleji- 
dar year this work has inclined; 

1„ Further iagorovement of the ultraviolet televlsicaa asicroEcope aM osei"= 
scope, synchronization of the coaiponents for tims -lapse clascBtography and a r&: 
erence calibration Btgja&l device for control of accuracy and raprodueibility. 

2. Development and application were conpleted of accurate cell counting 
and volume size distribution analysis of cell populations by using the jaodified 
Coulter Electronic Counter. 

3' Observation and photography of the UV absorption characteristics of 
normal and abnornal living cells to fiarther establish base lines for cyto- 
chemical techniques. 



Part B included 



308 



k^ Tlis systematic survey oi' properties aod react ioas of tetrazoliiim salts 
for use in stiudy of intraeellular reactions was contlned„ 

5o A method was devised for o^asuring reaction rates of certain euaiyne 
systems in living individual cells utilizing tetrazolium salts. 

Be METHODS 

Television Ultraviolet Mic ro scopy 

The retraining problems concerning synchronization of the control system 
components were solved with the assistance of Robert G Ileuhauser and Fo S. 
Velth of RCA Plants Lancaster Pennsylvahia and Gerald Vurek, InstruEiant 
Section, KflH A constant reference device for calibration of the TV cell 
measuring circuits was designed and included with the advice and assistance 
of one of the TV engineers at Walter Reed Medical Research Institute, ThcHSJas 
Keller This device permits switching in a calibration signal from ttine to 
tine to assure accurate and conslsttat asasursEagat characteristics of the elec- 
tronic circuits and to determine correction for any electrical drift or other 
changes This assures reproducabllity of the settings of the measuring circuits 
so that repeated measurements of densities are reliabJ.© 

The dark bars caused by misalignment of the caiBSra shutter the ultraviolet 
source fluctuations and kinescope framing time have been eliminated by proper 
alignmient of the operating components in the time lapse systern. The one re 
maining mechanical difficulty is the failure of commsr daily available shutters 
to provide reproducible open tiass within 10^. Conanercial shutters vary by as 
much as 25 ^^0^ and cause an annoying oscillation of picture density of se- 
quential franffis in time lapse motion pictxare records With the assistance of 
the Instrument Section we are attempting to design a shutter which will be repro 
ductble and provid® open tiaas within 10^ or less error. 

The amount of ultraviolet light in the band of 265 niji for clear photography 
by the television technique has been reduced to 10 milliseconds by further care- 
ful synchronizaticm and alignment of the electronic scanning, blanking, sensltls; 
ing light and signal amplification circuits Thus the selenium of the photocon 
ductive layer of the vldicon target is maintained at all tines diaring use in a 
sensitive state by illumination with low intensity white light to keep the elec 
tron "holes'" filled „ The instant before the ultraviolet shutter is opened this 
sensitizing light is blanlced off The storage properties of the vidieon are 
utilized by blanking the electron sweep beam diaring the open tin© of the ultra 
violet shutter then this beam is unblanked ai^ permitted to transmit the entire 
picture energy through the amplifier to the monitor during exactly one full fraii;£ 
period when the camera shutter is open for photography„ 

Several test motion picture films have been produced and Indicate satisfac- 
toiy alignment and operation of the system. Each fraas repi^aents 1 'lOO of a 
second exposiire of the living cell to ultraviolet light and at 1 picture psr 5 
minutes we have successfully photographed calls without apparent ultraviolet 
injury over a period of several hours. A satisfactory short motion pictui'e se- 
f^iVBizcm has been nede of the progress of raiuctloa of tctraaoliijin in liver aijd 



309 



oue culture cells o 

Cell Assay System 

The extreaie -wBriability ia the reaction of mouse ascites tumor cells, as 
determined "by reduction of tetraaolium farrmzsn, led us to try liver cells 
for such an assay system. We find that fresh liver cells, carefully prepared 
from perfused liver, by mincing, gently pressing through a series of fine-mesh 
stainless steel screens and final differential sedimentation in buffered cold 
culture Eiedia^ (Eagle's), provides nicely separated intact liver cells, without 
morphological evidence of injury 9 One of the earliest evidences of injury to 
cells is the swelling of mitochondria » In these cells, with both ultraviolet 
and phase microscopy, the mitochoiaSjria are very simll axA appear nonmlo This 
may be accepted as evidence of iainiiaal injury during the preparation procedure. 
We found that sows of the vBriabtlity in tetraEolium reduction by different 
batches of cells was due to the degree of o:^g8nation and therefor® elimiSKited 
the use of the buffer suspension izedia which requires COg - Pg sjeration. Eagle's 
or Hank's tissue culture media without glucose or neutral red are quite satis- 
factory. 

All individual cell studies have been controlled with test tubs experiments 
utilizing similar concentrations of tetraaolium in siAspeasions of several million 
cells per milliliter. To quantitate sreductions in these voIvesss it is necess- 
ary to accurately enumerate the cells. This was a^conplished by applying. the 
method developed and described last year, naaely counting and siaing the cells 
with the Coulter Electronic Co^mtero The work on evaluating this instrument 
and developing the method of coimting and volvass sizing large numbers of cells 
in suspensions, was completed ani the manuscript of the rex>ort has been submitted 
to the Journal of tl^ National Cancer Institute for publication o We now use this 
method I'outlnely to count all cell suspensions for tetraaolium ejsperiments o 

^trazolium Salts 

Further work has been continued on the purification of the tetrazolium 
salts, and Dr. Pierce has synthesisod several more with varying positions of 
the nitro and metho^ grovips to con^lete the series. 

Quartz Perf usion Chambers 

For detenaining reaction rates of individijal cells and iseasuring thera under 
the ultraviolet micrc^cops, a quartz perfusion chamber was desi^^d and fabricate: 
It has a depth of IOja, is covert with a quartz coverslip and pcssesses a 1 Esan 
thick quartz base. Around this chamber there is a deep moat, provided with tfe'o 
small Input and outflow tubes for continuous perfusion of fluids through the cham- 
ber. Yet the short depth of the chamber Eaintains the cells in petition, while 
perfusion introduces or changes the concentration of tetissoliun® or other reagents 
This has been successfully applied for titse-^^pse cinematography of progression 
of tetraaolium reduction by liver cells starting with no tetrazolium in the cham- 
ber, 

c. MAJOR nwmGs 

Modifications of TV-UV Micr oscope 

The major developmental modifications of the television equipasent which were 

. 310 



1<i 



Serial HOo KI-201 



being developed at the tims of the report for 1958 > have besa coiapletedo 
The synehronizatioa circuits and c^atroiki were conrpleted ajad continuing 
operation has been vary satisfactory, with the exception of tiss ultra- 
violet shutters, vhich break dotm frequently and req.uire repair. Tbm 
the system now permits tiioe- lapse photographs of either the loonltor 
screen or the oscilloscope GR Tube, utilizing ehoi't ultraviolet light 
flashits of 10 milliseconds in cycles iranging from 2 secc^ids to 1 hour. 
Our eaperienee indicates that different types of cells (kidjsey tissue cul- 
ture and ascites tusaor cells) vary n^rkedly in their laltraviolet seriBitivity^ 
This system provides a n®thod for accurate E^asureasent of the varying sensi- 
tivity of cells under different coalitiGOS. We are in the process of cali- 
brating oiir ultraviolet source, to enable calculation of ener^ ahsorbed in 
absolute imits. 

Continued work on the narkedly different reaction of individual cells 
and different batcl^s of ascites tumor cells has failed to elucidate the 
reason for this phenomena in tetras<^iuja r^uction. Liver cells are quite 
constant in their ability to induce tetrasolium from batch to batch anl 
■animal to anissilo There is soi^ variation between individual liver cells 
vhieh may be di^ to injury of preparation, but not nearly that found vith 
ascites tumor cells o Sise distribution curves of ascites tumor cells, 
particularly the Krebs type, at diffei?ent stages of growth from the third 
to tte twei^h day after innoenlation into aainsals, could not be correlated 
vith the varying reaction to tetraEalium» Sass difference was caused by the 
o^gen carbon dioside aeration of the si®p®Ming laDdium, but after this vas 
discontinued there was still a imrksd variability in reduction properties of 
different cells and batches. 

Preliminary experiments espies:^ the influence of several reagents on 
the volume of cells as measured by the eleetronie counter. The effect of 
chemical aad physical agents upon ascites tumor populations has been a sub- 
ject of intense research in recent years. Assay of these agsnts has been 
accomplished ^aerally by observing changes in population growth and bio- 
chemistry » SoE® investigators, however, have reported large changes in 
magnitx^ie of mean cell yoIubss and total cell number of treated tiimor popu- 
lations „ Conventional a^thods, such as optical sising or hematocrit procedures 
vere employ^o We B@re interested not on3^ in application of the electronic 
msthc^ to determining this chaoge in cell voluiiBe usder various influences, but 
also because this nsethcd permits the determination of the entire voluste dis- 
trlbutioa of a cell population aM thus wouM provide additional infornntioa 
concerning the sensitivity of iadividtuil cells in a given pcfp'olation, Krebs 
ascites carciniKia cells ^ere investigated. The effects of the following 
agents were sti:fii®d; 5 fluorouracii, nitromin, dinitrqphenol, hunan growth 
hormone (pituitaj^), plant polysaccharides (on carcoma 37), asEsd x-ray. In 
summary, the following table issdicates the changes in medican cell volume and 
total cell number caused by treatmsat of accvsrately counted ascites cell 
suspensions under the conditions of the e:^ariE©nts, %ese are 4stailed 
in a manuscript which is being submitted shortly to the Journal o^ the National 
Cancer Institute for publication. 



311 



i 



AGEKT m TOTAL CKi W, 


5 fluorouracil 


86,0 


ttltromin 


9O08 


2k mp 


80O 


105 g^nam growth honsiKie 


hl.k 


7000 gaHsoa groath hormone 


89a 


2k hotors post x-ray (8C0R) 


3.9 


kQ hours post s ray (80OR) 


2806 



BSCREASS IK MEDIM CEII, 

VOLUME 



16 o7 

58 »2 

2.7 

1J|-„1 

35 og 
22,7 
28 „2 



Osidatlon Reduction Reactions vlth T etragoliuia Salts in Ifviag Cells 

Studies have been coatlnued 1& the three toain categories reported last 
year that Is, the properties of the tetrasollum salts as lEdicatosrs^ their 
suttsbllity for investlgatloa of the biochemical significance of tstrasollum 
reduction to forusaaaa by these living cells bxA the a©chsais7a of this reduc- 
tion axil Its biolglcal significance. 

Fyope rtl es of Tetrasolium Salts 

The results of analysis of 2^ samples of substituted tetragoliuiss ware 
reported last year and this series has been continued irlth analysis of the 
purity and properties of 15 more synthetic derivatives (synthesized by Dr. 
pierce) o We hav® started the analyses for their properties, but results 
are not yet available „ Our previous flMln^ su^^sted aMitional positions 
for nitro and netho^ groups on the pl^i^l rln^« ¥© are lavestlsatiag tEelr 
properties to reduce sulfhydxyl compounds, reactivity with DPH diaphos^ses 
and their reactiv1.ty in dehydro^nase systems. It will be recalled that in 
our previous report we were able to divide these coaipouads into three ^neral 
classes those ^lich reacted with sulfhydiyl cong)Ound8 under certain conditions;, 
others which reacted with BPN diaphorases and del^ydrogssiase systeEoSo It was 
also reported that those tetrazoliuDB containing nitro gro'ops are the only ones 
reduced by the diaphorases and dehydrogenase systesiSo It was also reported 
that those tetrasoliurss containing nitro groirps are the cmly cases reduced by 
the diaphorases and dehydrogenase systess , but that the position and nuaiber of 
the nitro groups and possibly the position aM number of lasthGJQf groups deter- 
mine the reactivity 3?ate and perhaps specificity with different substrates ^ 
(succinate,, nalate etCe)o The additional confounds synthesized by Dr. Pierce 
completes the series of possible positions and nusft^ers cf nitro aai ssthosy 
groT^s and shoiald enable us to conclx;de this phase of our was-k. 



312 



E«rial No. liCI-201 



Although, uasbed rat and mouse liver mitoebondria possess a variety 
of deliydrogenases as determined by react iosas with different substrates, 
it was found that whole washed liver cells did not necessarily react 
simLlarly, There are several factors which may account for these differ- 
ences, es manifested, for instance, by one tetrazoli vim which reacted very 
rapidly with washed mitochondria, but very slowly aM with much less inten- 
sity in liver cells. deDuve, in Belgium, has pointed out that it is impossi- 
ble to remove mitochondria from cells without producing a considerable injury 
and probably a shift in ensyme con^sosition and co-enzyme aM inhibitor activi- 
ties i Thus liver mitochondria are disoriented aM tizKloubtedly injured to soae 
extent when used in these assay systems. On the other hand, their orientation 
remains relatively nonml when whole cells are used in suspensions with cultux^ 
fluids „ Therefore s we are repeating all the work with tetrazolium dehydrogen- 
ase systems and several substrates in whole liver cells, utilizing tl^ iiltra- 
violet microscope for imeasuring reaction rates. Thus far, marked differences 
in reaction rates have been found for several tetrasolitaa salts in individual 
liver cell systems. 

The question arises concerning the cavise of the difference in reaction 
rates of the various tetrasolium compoviJsis. It may be due to differences 
in cell jwsrmeability. We have succeeded in measuring the rate of permeability 
for several tetrasoliuns o When the cells are incubated in tetrazolium only, 
for i>eriods of 15 to 30 minutes, the addition of a rapidly reducing chemical 
((HHi^)2 SO3) there results immediate reduction of the tetj^azolium which is 
present in the cytoplasm. With the few tetrasoliums tested so far^ there seems 
to be no difference in perasability i^te. When the cells are washed several 
ttniss after incubation with tetraaolium, the washings fail to wash out more 
than a slight amount of the tetrazoliuin. Completion of these e^cperisosnts 
should provide conclusive data concerning the perneability rate of the differ- 
ent tetrazoliuffis, and the influence of pemaeability on the enzym reduction 
rates of the tetrazolitms . 

It was fovind also that concentrations of tetrazolium and substrates 
(0 .0012M tetrasolium and .03^ succinate) result in rapid intense reduction 
in the cytoplasm of the liver cells, apparently? in the vacinity of the mito- 
chondria o There is a general increase in density to ultraviolet throughout the 
cytoplasm^ However j, when much lasher concentrations of both tetrazolium sz^ sub- 
strate are utilized there is a slofw reduction to formazan, vary little or no eb- 
sorbing fomazan can be observed in the cytoplasm, but the fat droplets grsdiiall.y 
darken. This apparently indicates a transfer of the lipid soluble formasan as 
it is formed to fat droplets before sufficient can accumulate in the vicinity 
of the mitochondria to be detectable even by UV microscopy o We hope to synths- 
sise a fat insoluble formasan to avoid this phenoEsnon, 

Biochemical Significance of Tetraao lium Redu ction ty Living CellB^ 

Stiidies have been continued in an attempt to elucidate the causes of reduc- 
tion of non-nitro tetrazoliums by living cells, since tlsse co!!?>ounds are not 
reduced by the homogenate systems. The findings i*eported last year of nitro 
conpounds being poorly reduced when glucose is supplied to the cells andg on 
the contrary, the non-nitro compounds, after a lag phase being readily r^uced. 



31^3 



e conflmffld; but ftirther elucidation of these pi'ocesses was interrupted 
..^ the permeability ei^erisseats c We decided that persaeability, the taore 
likely cause of differential reduction^ should be studied before more coiHp- 
licated mechanisms of reactions within the cells ^, We have also confirmed 
our previous observation that although liver cells aini certain tissue culture 
cells do not spontaneously reduce tetrazoliums in the absence of added sub- 
strates, such as succinate or glucose, the ascites tumor cells of mice in- 
variably reduce nitro tetrasolixais without substrates o This difference in 
reaction between normal liver cells "aol" ascites tumor cells merits further 
study to determine the mechanism involved. 

Attempts to quant itate the reaction of tetrasolixm reduction in living 
cells have been successful „ Ifeasurements of the general and specific absorp- 
tion of liver cells, at 3000 AO, were made beftsre and at intervals of 2^, 
5- 10, 15 and 30 minutes after perfusion of th(j cell chamber with the proper 
concentration of t^trazolium and substrates. 'i?he increasir^ density of the 
cytoplasm due to the reduction of the tetrazollum in fine particles at the 
sites of the mitochondria ¥ere measured by the single line tracing on the 
oscilloscope and the density calculated from th(j voltage readings. On the 
tracing, the voltage from the base line to the ^rhite level (total trans- 
mitted light surrouTKling the cell), represents total Incident ultraviolet 
light, and the voltage reading between the base line aai the deflection of 
the cell cytoplasm represents the light transmitted through the cell, A log 
of the z-atio of the first to the second voltage rsaditjgs eonstitirtes the 
density of the cell cytoplasm for the light path Involved, (lOji). This was 
measured and calciilated for the sane portiim of tfce cell at the time inter- 
vals mentioned o From these data, reaction rate cui'ves were calculated and 
plotted for several tetrasolium-substrate combinations „ This work x^as re- 
ported at the Cytochemistry Conference in Copei:^ageEi in Apx*il and subsequent- 
ly was submitted for publication in Esperlmantal Cell. Research, (next supple- 
muntal issue)., 

Do signtpicam:® to the mtioml cPMsm iKsrinnss 

It has been sho^n that qimntitative data of reaction rates of an enzyss 
system causing the reduction of tetrazoliiim to fonmsan can be measured over 
periods of time in living cells for the purposes of acetmsy-latiag infors^ticn 
about dynamic changes in setabolic activities with a miniEum change in the 
environiBant of the living cell system. This type of study utilising ultraviolet 
television microscopy and time-lapse cinematography for determination of reac 
tion rates in living intact cells will contribute to our knarledge of the chemi- 
cal systems therein a We propose to apply these methods to tlie study of chenical 
systems in normal aM neoplastic cells aM their responses tct a^nts such as 
chemical inhibitors and irradiation^ 

Eo PRQPOSatt COURSS 

lo Studies will be continued to elucidate the mechanism of biological 
reduction of several different tetrasolitan conrpoujsis, their possible specific- 
ities in living cells, aM their nanifestatlon of cell nstabolic systeisso 



31i| 



2. Patterns of tetrasoli'oia reduction aad progressive changes in 
transfer of the fomassaa from laltochorairlal areas in living c5e3J.s to fat 
droplets, 'srill be i"®corded with tlme-lapBe photographs in a continuing 
study of these localisations in different types of cells, inclxading liver 
cells ; kidaey cells, tissue culture ceLls and ascites tumor cells » 

3o The shortest exposures possible to ultraviolet lights that Is 10 
millisecorsds at varying intervals, still produces dainage after accumulated 
expofiures, aM is visibly itEaaifested by swelling of the mltoehoodrla, swelling 
of the cell cytoplasm, (probably by iidslbltion of water), aM finally produc- 
tion of cytoplasmic protrusions (blebs), through the cell wallo Several in- 
vestigators have reported that ultraviolet damage of yeast cells aM soase bac- 
teria is reversible by longer wave lengbhs of light o We propose to interpose 
a white light source In the UV raicroscope sjretem to irradiate the cells with 
different wave lengths and periods of white light in an atteispt to reverse the 
ultraviolet dainage produced by the short ejtposures to xiltra violet at 265 and 
300 mji. 



315 



Serial Eoo 201 
Office of Associate Director 
in Clias'ge of Research 
national Caneer Institute 
fethesda^ Maryland 



Iiidi'vldiJal Project Report 
Calendar lear 1959 



Part_Bo Honors^ Awards^ asid Pi&lieations 

Pi^li cations other •fchaa abstracts frran tMs project; 



¥illisas^ So Z.s K.ffis Lapse Ultraviolet Television MLcroecqpy 
Instruaaentation and Biological Applications <, Ire Transa.ction@ 
on tfedical Blectroirf.cgj, Volioae M-6g6S»7i4-^ June 1959 o 



k 



31G 



Serial 



JJCI-216 

1. NCI 

2. OADB 

3. Bethesda 



PES - NIH 
Individual Pinoject Report 
Calendar Year 1959 



Part A 



Project Title ; Drug Develoianent and BvfiLLuation Program 

Contract No. SA-l*3-ph-2371 
Project Officer : A. Goldin 
Assistant Project Officer ; J. M. Venditti 
Assistant Project Officer (Adm) ; G. A. Brsndner 
Contractor ; Microbiological Associates, Inc. 

St. Invest i^tor ; I, Kline (l/s time) 



gun Time Personnel ; 

E. W. Artis 

H. Waters 

L. Mitcliell 

I. Hughes 

P. Freedenberg 

C. Jackmon 
L. Keys 

G. Sanders 
E. McCullo\2gb 

D. Scott, Jr. 
L. Williams 
M. Washington 
A. Slau^ter 
Wo Todd 

E. Gray 
J. Keys 

J. Mcllvain 
R. Wheeler 



Crev Chief 
Ass't Crew Chief 
Sr. Chemist 
Biometrician 
Secretarial SerH,ee 
Sr. TechsAcisn 
Sr. Technician 
Sr. Technician 
Sr. Technician 
Sr. Technician 
Sr. Technician 
Sr. Technician 
Jr. Techsdcian 
Jr. Technician 
Animal Caretaker 
Animal Caretaker 
Animal Caretaker 
Animal Ceuretaker 



Fart B Included Yes X 



317 



SCI 216 

Part A: (cont lamed) 

Calendar Year 1959 

Page 2 



C. Aulisio Crew Chief 

C. Brown Ass't Crew Chief 

E. Philip Technician 

C. Lester Technician 

J. Exvec Technician 

W. Cttrtis Technician 

H. Coleman An-tmal Caretaker 



I 



31S 



mi 216 

Part A: (coatinued) 

CaleMar Year 1959 

Page 3 

Cooperating Ualts ; 

(a) Laboratozy of Cheaaieal Phasaiacology, NS2 

(b) General Medicine Branch, HCI 

(c) Cancer Chemotlierapy Eatioxsal Service Center, 
^•iiich supplied laost of the cosrpounds ess^loyed 
in these studies 

Man Years (Calendar Yea: ,^^2* 

Total: 20 (adjusted for lea^h of service aad part 
t^jEs crew) 

Professional: 2 

Other: l8 
P roject Descr iption; 

Preel^jaieal drug evaluation and developsestfe 
^thods Bnaployed: 

Methods established for quantitative drug evaluation 

in the Laboratory of Cheaical Pha3raiacolo^^ 1C2. 

Otl^r applicable procedures. Standard pharsiacologieal 

procsdiares. 

jfejor studies: 

In accordance with the recoasn^ndatioas of the Associate Direc- 
tor in Charge of Research and the Clinical Director, HCI, the drug 
evaluation and develoxa^nt program has been in progress for the 
farther stisdgr of coEspounds of potential clinical interest ineltidlns 
(a) selection of drugs for clinical trial, (b) obtainiEg of basic 
end practical infonuation coaeeraing such drugs, (c) referral of 
specific clinical problems 'KasTanting animal experinsatation. The 
program is being conducted by Dr. A. Goldin as Project Officer, 
J. M. Veaditti as Assistant Project Officer, and G. A. Braafiner as 
Assistant Psroject Officer (Aam)^ on contract with Microbiological 
Associates, Inc. The Project Officer is responsible for the paro- 
graa directly to the Clinical Director, KCI, and the Associate 
Director in Charge :>f Research^ NCI. Close liaison is laaintained 
with the General Medicine Bi^nch, the i^iboratoiy of Chssiical 
Pharmaeolo^, and other groups in the intraiaural prograsu. The 
program is sxm. in close eooardination tfith the Caijcer Gheraotherapy 
National Service Ceater. 



319 



NCI 216 

Part A; (co&tinued) 

Calendar Year 19^9 

Page k 

•Else Bsthodologies onployed for the quantitative evaluation of 
agents in experimental cancer have been developed primarily in the 
Labosratoxy of Cheaical PhartKicology. Additional laethods are being 
developed in the IsiboT&toT^ of Chemical Phsratacology sad in this 
prograai. 

Tbree laouse tuaors have been esployed in the general evalua- 
tion of new agents of interest: Leukeiaia (L-ISIO), sarcor^ (S-37), 
and earciiKKaa (Ca-755). Preliminary studies are being conducted 
esiplc^ing t^e Shrlich carcinoma, hepatoE^ 129, sarcoma 16O, and 
other ezperiiaental tusiors, including virus induced tisoors, in order 
to detexmiQe the fsasability of incorporating additional test sys- 
te!33 into the program. 

With the advent of active compounds of different chemical 
classes, drug resistant tutsos^ (sntifclic resistant tiasors, anti» 
pmlne sreeistant tusors, etc.) are being utilised to exploxe poss- 
ible i^ehanisms of drug action. In addition, certain active com- 
pounds have been subjected to detailed studies involving the de- 
texmination of optissal seheditles of treatment, opting route of 
administii^tion, the effects of trealsaent vith combinations of 
drugs, etc. 

A. Studies with Leukemia L-1210 ; 

1. Eo^loying Ametbopterin as a standard for sntileukssiic 
activity, 73 additional eo^^unds were tested on subcut- 
aneous trea^Ksnt over a wide range of daily doses against 
advanced leukesaia L-3210. ^e coi5pounds ^ere rated rel- 
ative to Aisethopterin T^ith i«si«ct to their ability to in- 
crease the stu^val tii^ of mice vith advanced L-1210. 
IQie structiare-antileukesiie activity relationships among 
these compounds n^iy be si2SEDmrized as follotm: 



o 



20 





HCI 216 




Part A: (continued) 




Calendar Year 1959 




Page 5 




Relative increase in median 


CcxBpoTmd 


stzrvival time over controls 


at the optiiaal daily dose. 




A^thopterin increase e 100 


a. Alkylating agents 




C^rtoxea 


97 


2-/Bis (S-chloroetbjl }amiiios 




laathy^-J, 6"diiafitliylbea2i- 




midazole 


35 


Glyeizte nitro^n smstard 


23, 18 


meta Sareolysin 


19 


Alaoiae nitrogen laustax^d 


13, 11 


BydrpqialDotm imstard 


12, 11, 10 


g.^Bls C2-chloroetli^l)e!Binos 




nuatb^^obeuziMdasol® 


7* 


H'^,I!I'^-Bi8C2-chlo^oeti3yl)-l3' - 




2-pjprid3rlsi^Tfanilamide 


7* 


p-H,H-/Bi8 C2"iodoethyl)amiBos 




bsazyl/»piiosphonic acid. 




diethyl ester 


6 


ChlozB^ttcil 


6 


p-/Bi8 C2»chloroetl^l}Effiino/e 




benzoic acid 


k, 


Hy R-Bis (2- chloroethyl ) stilf anilie 


acid hydrazide 





^lez'an 





b. Antibiotics 




Streptovitacin A 


ko 


Actidione 


26, 25 


m-lkk 


29, 


Aetino!£(yein 3 


k 



c. Cyclopentane carbo^grlates 

Me«jyl-l-aaino<^clopentaa8-l= 
caxbo^late 



38 t 



321 



I 



NCI 216 

Part A: (contimied) 

Calendar Yeor 1959 

Page 6 



n-Btt^l-l-aminoq^lopenfcane-1- 

caz^xylate 33 t 

n-Propyl-l-aminocyclopeatane-l- 

cajrbos^late 33 t 

1-Ai^jao^elopentane carboxylic acid 33, 27 

Isobut^l-l^sQBinocyclopeataQe-l- 

carbosylate 28 t 

d. Bieotialc acid derivatives 

6»Chloronicotinic acid 4- 

Nieotix^mideoH-oxide k 



Metbylslyoxalbisguao^Uurdx^zone 

dUQTdrochloride looxKifaTdrate 68 
ifet^lgl^Kalbisguaoyliiydrasone 

diaeetate tril^pdrate 62 
Ifetbylgl^xalbisgimDylhj^drasoiie 

iQoaosulfate mooohiydrate ^6 
GlToaEalbisguaayllSiydrazone moooa 

sulfate pentahydrate 9 

Furljse derivatives 

(6-Mereaptopurine vas included in all €:q)sri]^snts involving 
the testing of new purine derivatives. On the avera^, 6- 
lisircaptopurine is aboHt 50 percent as effective as AE^toopteria 
against advanced L<>1210). 

6«1liiogi2anine 50* ^ 

6-lbiogisanine rilsoside 54 

6-Clfetlvlthio)purine 12, 8, 8, 7 

6-(Etliylthio)piiarine 7 

6-(n-Prop3rlthio])puri3Ki 55 > 23 

6-{l8opropyltMo)purine 55* 37 

6-(n-»it^lthio)purine 3^* 2U 

6-(l8otoitylthio)purine 31 

6» (eee- Batylthio )purine 38 

6-(n-Penfcyltbio}purine 39, 2k 

6-(n-Hex3rltbio)purine 



322 



HOI 216 

Pari; A: (coatiQued) 

Caleadax Year 1959 

Page 7 

6-(a-Eeptgrltbio)pairine 

6-(B-0c1^1thio)pui?lne 

6~(A13ylthio)purine 26 

2-AmiiK>-(6-'b€nsylthio)purlne 2h 

6-(2-(2iloroal3ylthio)purine 26 

6-(o-Cblorob€nzylthio)purlEe 2 t 

6-(2°Chlorobsn2ylthio)purlB8 k t 

6oC3en^Isastl3srlthio)piarixie 

9-Eth3rl°(6-iset!syithlo)pmlBe 

8<>Msftl)3rlthlo>6-pmli^thiol 

g. P^razolo pyrimldlnes 

(The effectiveness of U-AsstaoKrrazoloCSj^-dll^^miSiBe has 
been Bbava to raii^ fross 19 to 33 percent of the Metbopterin 
efftect) 

k" (n-Propylaaaino }^rasolo ( 3^ ^-d}a 

X^risldine 29 

lf-^Bsylaaino|^i^ffiolo C 3, ^-d )s 

^Tifflidlne 29 

k" C 3, 2(.«.sichloK>ben^laiBlno ) j^i^solo ( 3 , l*-d }s 

pjniMdine 10 

l-MetlQrl-^-H-^ifcylaiaiaopyrssolo (3, ^-d)s 

pyrlEsidine 7 

h. lalogeaated pyrimidines 

5-Fltsoroorotie acid 5^ 

5-KLiK)roaracil 39» 39 

3-Fluo»>deoKyuridine 39? 35 

5»Chlozoaracil 6 

5-BrDQK>uz«eil 

i. Other j^rimidines 

6-A3aiaracil riboside 13 t 

6->Asauraell 6 t 

5-Diazouraeil 



323 



Hd 216 

Paart A: (coatisued) 

(^leadar Year 1959 



J. Colchicioe derivatives 

N-Acet^rlcolchinol; oetlayl ether 11 

Q-Ace^lcolchisol 7 

Coldtiiiiol 

k. Stgrzylqulnolines 

^•*(£-Di®t6orlsminostysyl)<j^aQliiie 6 
l^.(j9»Di!setl^leiaiBostyz7l)QuiJK}lise di HCl 6 
S- (2-SliBetlqrlsaiao8tyx7l}galsolin® 

laethiodlde 6, 

k' (^-DietljylaiBinoslyzyl ) quirpJ-ln® HCl 
k» (^^-MiaethylamlBoatycyllmtiBnl-lBa 

(free base} 
I^. (^oDisetlqrleminos-i^iyl ) quinolias 

nsethiodide 

1. Others 

£*£*-DDD 9 

Potassitsa oor^olizto oKssi^te 8 

2-listhyl-DL-Borleucin© 

DLoS*AMQO'>2'*ISsthyl«ll-dodecasoic acid 

Galactoflavia 

Hareotias 

tolbutamide 

2,U,7-TriaEiino-6-o-tolylpteridine 

^ or t isidieates that the optimal dose jb^ have been outside the 
raage essplo^ped. Inhere all dose levels prodused toxieitj^ denoted 
"by ^, the optisaal dose nsay be lover than a^ ^aplc^d. Where no 
clear toxicity appeared at sss^ dose level, demoted "b^ t, the opt« 
Imal dose la^ be higher thaa a^ esirploj^d. 

(In addition to the 78 cc^spotaads tested a^djist L-ISIO for l&e 
^rst tins in this programs vithin the last year^ 10 com^ym&s 
first tested ia the previotis ^ear ^lere ree^aiainM in this asss^ 
S3 



321* 



NCI 216 

Part A: (contimxed) 

CaleMar Ye&r 19^9 

Page 9 

2. Special Stttdiee CL°lglO) ; 

In addition to the aforen^entiosed ciiHitparisons of antile^^sic 
effectiveness on daily suS>cutaneoY2s treatzssnt, certain cosapounds 
\^re suibjected to farther studies as follows: 

a. Stisdies on the inflfjience of the schedule of treateeent on 
antileuk^tdc effectiveoesss ohonsd that for Mitoii^cin C, 
Streptovitaein A^ AetiiK»^cin J>, E°73 acetate, sad t&3 
isetb^lglyoxalbiBSiiaa^lhsrSrasones, dai]^ treatsssnt ^^as 
more effective than treatments at ^der spaced intervals. 
In contrast^ CSrtoscan displaced progressively greater 
astiletzksaic effectiveness as the interval "bet^^en treat° 
iBsnts vas extended frcua one day to one neek. S'^S"" 
diehloroassethoptesrin displayed a s^rked degree of anti» 
leukeisic effectiveness over a vide range of treatsient 
schedules from t^ce daily to once evexy foor d^rs* ISie 
letter result is in contast to the results x^ith As^tbs 
opterin ^hich vas relatively ineffective against advanced 
L<>1210 ^en given evesy four days. 

b. Studies of the influence of the route of drug administration 
on antilei^^Bic effectiveness showed that 3'»5'-'^chloro8 
QSiethopterin and AEethopterln .were more effiective ^en given 
parentei^lly th^i i^en given orally. 12he reduction in the 
effiectiveness of 3'*5'-fiichloroaj^thoptsrin vfeen the drug 
vs^ given orally ^fas sufficiently extensive to res^ilt in 

an aatileuk^aic activity no greater ths^ that ol^tained 
«ith Aioethopterin. IMs demonstrates the importance of 
studies of the inf^ne^ce of the rout® of adadnistration 
on the effectiveness of drugs and sug^sts that in this 
program greater ^aphasia be placed on such studies. 

Further studies of the influence of the route of e4Mnls» 
tration on the effectiveness of N-KSthyl-N-nitroso-H« 
nitroguanidine against advanced L°-1210 sho^nsd that this com^ 
poimd vas ineffective l;^th parenterally and orally. 

Studies of the infli^nce of intravenous aSiainistx^tion of 
cer^in coiapounds are in progress. 



325 



HCI 216 

Part A: (contimstsd) 
Calendar Year 19^9 
Pass 10 

c. Studies of comblaation and altenaatizis thez^s^ against 
advanced L-1210. 

Concomitant treats^tnt of mic@ Hith advanced L°1S10 ^th 
Cytoxan and Amethopterin or daily treatnieats alterna- 
ting Tieekly between tliese drugs x^sislted in little or 
no advantage over daily treats^xst ^th either drug alone. 
Bovaver^ a single treatesent vith Cytoxan fblloved hy 
^teily treati^nt vitb Araetbopteria ^ms considerably mare 
effective tban daily treatistent ^th AsaeUiopterin alone. 
Of particular intex^st vbs the observation that treat- 
laent every k ds^ ^th AESsthopterin preceded by a single 
treatiaent vith Cytoxan had a loarked ef£)eet agiainst advanced 
L»1210 ^^hile treat!!3ent evesy four d^s vith Aoethopterin 
alone Tfas relatively ineffective. 

I^ily treateent altematisg ^i@ek3y bet^fsen ^isst^pterin 
and 6«>t^rcaptopurine was more effective than contismo^a 
treatiBent vith either drug alone. 

d. Studies of the relative effectiveness of G^ftoxan and 
AiBSthopterin a^inst early L-1210 shoved that C^^^can 
displayed Hi® capaei^ to produce tumor free survivors 
vhen given as a single treati^nt vhereas Amethopterin 
did not. 

e. Investigations of the influence of sex on the aatileuk- 
effiic effectiveness of Cytoxan showed no diffex-enee in 
response to this drug in male and fszisle leu&esaic laice. 

f . Treatssient of jsoxmal mice in the sssas esqperiiaeHrb ^^ith 
let&emic laiee shoised that the tonicity of Qftosisa for 
the host liM.ts its therapeutic usefalasss to a greater 
degree than does ^e host toxicity resulting from As»stl^ 
opterin trea'toent. 

g. Boat toxicity due to 3',5'-dichloroaiaethopterln limited 
its therapeutic usefulness to a greater degree on oral 
administration than on sv&cutaneous administration. 



326 



SCI 216 

Pari; A: (continued) 
Caleaoiar Year 1^9 
Pa^s 11 

@i9 drug displa^^ approKimatel^ the sazoe to^eie effects 
OQ noxioal mice ^iSsen givea orall;^ or s^cutaaeousl^^ but 
the antitumor effiset ^ms maxkisdljr rsdncsd on oral ad- 
ministration. On oral adiainistration ^tdi Sichloros 
cffiiethoptei^ii izihiMtOB of th@ local tiassor v;;^ is^mri- 
ebl^ associated ^tii drag toxicity for the host. 

B. Stipes td,th drug resistant variants of L-ISIO: 



Studies of the efftaetiveness of U-^aiEop3?rasoloC3»^-d)e 
Xigfriaidise against a subline of IioI210 resisteat to i^oth 6- 
Ifercaptopuriae and &>Azaisusnine Bhsws& tMt t&ts ^rasolos 
pyrindfiiae ^jas able to elicit extensive reductions is local 
tUEior size hut vas z^letively ineffective in increasing the 
survival tirae of the mice, ^iis suggests partial cross- 
resistance bet^sen 6-Mercaptopurine and 'i-Aai3o;^rx€zolDC3^4-d)s 
PSTidMdine. Additional studies ^Tith the 6-AzaguaninS"6<-Bier^ 
captopurins resistant v^^ant of L-lglO showed that it «as 
resistant to treatiaent ^th 6-l^rcaptopuriBe riboside, 6- 
AU^lthiopurine, 6-C2«»Chloroall^ltbio}purine, 6-(n-Butjlthio)s 
lAirins^ and 6-Clsopropyithio)p«ria® but sensitive to Cytoxan 
and ^methopterin. 

Studies ^d.th sa Assethoptesln resistant s^lise of LolSlO 
sho^ied tdiat this variant isas B&smltlvQ to treatiaent vith Q^oxan. 

Studies of the effect of cosbiaatioa treatsent ¥ith 6- 
Msreaptopurine and Azasezls^ in Mce bearing the S«Azagu@Mss- 
6-Hercaptopuzlne resist^it variant of 1>1210 suggested that the 
coc&ination «as no ^}re effective th^ treats^nt tfith Aseserise 
alons. Against the sensitive L-1210, the coisbination ^as incre 
effective "Qian either drug alone. She synexgistic effect ises 
ha^&veTf on 'Uie schedule of treatoient e^lojed. 




1. Biirty-five coiapounds ^ere studied vith resi^ct to their 
abili^ to extend the survival tioie of ndce vith relative-' 
3y ear3jr S-37. In genexml, treatsssats ^?ere tegoa three 
da^ after tuaor inoculation aad give^ daily for 5 d^s. 



327 



mi 216 

Part A: (costi^ued) 
Calen^r Yesr 1959 
Page IS 

Each coiopouad vas odisiBietered over a t^d® razigs of doses 
and the median survi.val time at the optimal doss of each 
e<»Bpou3ad vas ccsoparsd vith the median smvival tisjs of 
the untreated controls. Compounds providing inc2?eases 
in survival tlice over controls of 40^ or less ars grouped 
together and considered inactive in this report. For 
purposes of coisparlsoBy the effects on the local t^aaor 
of S-37 are shown. 

a. Conrpounds producing a greater than i^O^ iahiMtion of 
the local tiMor at dose levels resulting in not ox^re 
than 20^ lethal toxicity on the tenth da^r after tUBur 
inoculation (three d^s afber final treatiisant) and 
providing increases in s^dian siarvlval tlmB of at 
least kO^ over untreated controls at the opti^U. dose. 

Alanix^ nitsrogsn mustard 

@l^cine nitrogen oustard 

Carzinojdsilin 

5-^uorod903iynridine __ 

p-/BisC2»chloroetl3yl)aiaiao/bensoic acid 

Azaserine 

Chloraisilmcil. 

"bo Cosipounds producing a greater than 1*0^ inhibition of 
the local tuzaor hut f&iling to provide a kO% increase 
in isedi^i survival time over controls. 

Actinom^in D 
Bayer 323I » 
Q^toxan 

Sareo3ysin (DL) 
5-Fluoroorotic acid * 

(■SMice succuB&ed to toxicity over the entire dose 
irasge. Tbs possibility rernains that t^se ccxspounds 
might provide greater increases in msdi&a survival 
time at lo^jsr dose levels). 



328 



mi 216 

Part A: (continued) 
Calendar Year I959 
Page 13 

Compounds which neither inhibited the local tussor by 
40^ nor provided more than 1^0^ increase in median 
survival tiias. 

Actidione 

Miton^cin C 

CB IJHU 

Degranol 

E-39 

Eydroqulnone mustard 

MEIPA 

OPSPA 

TEPA 

■TEM 

6-Azathyinlne 

1-Aminocyclopentaae carboicylic acid 

Glyoxalbisguanyliiardrazone and 3 of its methylated 

derivatives 
Colchinol 
N-Acetylcolchinol 
N-Acetylcolchinol, methyl ether 
Hyleran 
Benz imidazole 
9-Chloro'»9- C^-methylthioxyphenyl ) -lO-pheoyl 

acridan 
o,p'-DDD. 



2. Special Studies 



a. Studies of the influence of the duration of treatiaent on 
the effectiveness of 5-Fluoro\iracil and Carzinophilin 
against S-37 a?® in progress. 

b. N-Methylfonaamide was tested against various tv^ssr inoc- 
ulum levels of S-37 in order to detei^i^ its capacity 

to produce tumor free survivors. Trea-feaent '^th N-Met^l^ 
formamide did not produce an appreciable niuab@r of tumor 
tree survivors. At low inoculum levels, the number of 
"no takes" in treated groups was not sigaificsatly greater 



329 



mi 216 

Par-t A: (continued) 
Calendar Year 1959 
Page 14 

than the number in the untreated controls. Similar 
studies are in progress employing Alanine nitrogen 
mustard. 

c. Studies of the infl.uence of the route of administration 
on the effectiveness of 5-Fluorouracil against S-37 in- 
dicate that the drug shows about the saiae activity ^en 
given intravenously or subeutaaeously. 

d. T&e relative insensitivity of S-37 to therapy, mates it 
a tuiKjr of particular interest for further study. 

D. Studies with AdeaocareincHaa CCa°755) : 

1. It had been observed that 6-Mercaptopurine provided a 

iBarked effect against Ca-755; producixig not only a high 
degree of tusior inhibition but also extensive increases 
in survival time as^ a certain nijiaber of tuaajr free 
siarvivors ^en treatsient ^/as begun early (day k-) and 
continued for 5 days. In the past year, U5 coaixiunds 
were tested over a wide range of doses employing this 
schedule of treataient. Eie relative effectiveness of 
these compounds at their optimal dose levels in extend- 
ing the survival time of "Uie mice is suasaarized as follo^?rs; 



Relative increase in median 
Compound ' survival tia e. CoatrolB s 3 

Puisne derivatives 



Average l^i- 



6-Mercaptopurine (dissolved 


in 




dil KaOH) 




( Ai 


6-Mercaptopurine ( suspended 


in 


methylcellulose ) 


159 J ' 


6-MP riboside 




223, 233 


6-^ioguaaine 




203, 193 


6-Thioguanine riboside 




173, 211^ 


6- (n-Propylthio ) purine 




169 


6-Chloropurine 




163, 165 


6- (Benzylmethylthio )purine 




161 



o 



30 



KCI 216 

Part A: (cozitinued) 
Calendar Year 1959 
Page 15 



6- (Benzylthio ) purine 152 

8"Ainino-C6-benzylthio)purine 152 

6-E^iylthiopurine IU6 

6-(o-Cblorobensylthio)p«rine lUl 

6-lfethylaiercaptopurlne I3I, 123 

9-Ethyl-6-metl3yl'bhiopurine 229 

6-C2-Chloroallylthio)piirine 126 

6-Cn-P®ntylthio)pTirine 122 

6- (n-Eeptylthio )piirlne II6 

9- C2-Cyclohexene-l->yl)-6-piirinethiol Ilk 

8-Methyl-6-purinethiol 100 

b. I^razolo pyriiEdines 

l^-n-ProIgrla^ninopyrazolo C3p*^-d)» 

pyrimidine 125 

U-ABinoxgri'azolo(3,'^°d)pyrii!iidine 117 
U-Ben2ylaminopyra2oloC3**4-°d)pyriraidine HO 

c. Halogenated pyrimidii^s 

5-Fluorodeo2{3ruridine 12^ 

5-Fl\K>ro«racil 121 

5-BrcM!so\arecil 103 

5-(2iloK)uracil IO3 

d. Other pyrimidines 

2,U-Die!aino-5-Cl*' ''Chlorophenyl)-6- 

etl)yl pyrimidine IO8 

6-Azauracil 119 

e. Alkylating agents 

Cytoxan 195, 150 

A-139 117 

Thio-TSPA U2 

nitrogen mustard IO8 

f. Antibiotics 

Mitomycia C 327 



001 



ECI 216 

Part A: (coatiaued) 
Calandar Year 1959 
Pagss 16 

StreptoTitecin A II5, 112 

Actidione II5, 132, 108 

Actinos^ein D 10^ 

g. Others 

l-Aaitjo^clopentane carboxylie acid, iteethopterin, ^ydroa 
cortisone. Prednisolone, aM o,jo' -WD, aM four gl^xalblss 
gtaai^Utydrazozies failed to increase the lifetime of mice idth 
Ca-755. 

Of particular interest in the stitdies of drug effects against Ca- 
75? ^»Bs the oliservation that coigcpoxinds may markedly inhibit the 
local tvsEor without appreciably increasing the survival time of ths 
mice. For e^caiaple, all of the pyrazolopyriialdines listed s^vq 
produced approximately ^0 percent inhibition of the local ttssKir 
diameter at dosage levels caissing lethal toxicity. At optiical 
dose levels, however, the increases in survival tiate provided by 
the ^rasolopyrimidines were relatively small. 



Because of the gratif^ng results of the early treat- 
ment of Ca-755 with saay purine derivatives aad Cytoxan, 
a study is being zaad® of the feasability of ez^ploylng 
ih±a tu^>r in a more advanced form as a test for aati- 
ttsaor activity. It was shown that 6-MerGaptoptn"ine pro- 
duced a 79 percent ia median survival tii^ over controls 
T&en 5 daily treatsjsnts wer^ begun ei^t days af1;er tuaior 
inoculation as craapared with a 137 percent increase lihen 
treatiaent ■was begun four days after tuaor im>eulation. 
Cytoxan provided an Bk percent increase in survival tiaie 
against the advanced tismor and a 95 percent increase 
against the early tusjor. 

In each experiment involving comparisons of drug effects 
against Ca-755, 6-lilercaptopurine was used as a standard 
for antitumor activity. Depending oa the vehicle em- 
ployed for the test compound, 6-Mercaptopuriae was either 



332 



NCI 216 

Part A: (contisusd) 
Calendar Year 1959 
Page 17 

dissolved in dilute sodium hsrdro3d.de or siaspended in 
methylceUulose. In the course of the studies^ it vas 
observed that 6-Msrcapt@purine was wore effective in 
dilute sodium hydroxide than ia setl^lcellulose asA that 
the compound vtsm more toxic ia suspension in ssthylcellu^ 
lose. In order to investi^te the possibility that thi^ 
difference could be due to differences in absorption rate, 
a study of the relative effectiveness of 6-Mereaptopuriae 
in the tijo vehicles over varioi£s treatment schedules is 
being conducted. 

C. Daily treataaeat ftor five days of early Ca»755 t^lth Adeaylic 
acid and 6-Mercaptopurine or 6>Mercaptopurine riboside 
showed that Adenylic acid failed to diminish the antit^^u^r 
effectiveness of loereaptopurine or its riboside. 1!he 
possibility that Adenylic acid coiO-d block the effective- 
ness of a single treatment vith 6-Mercaptopurine is under 
investigation. 

B. Studies with Ehrlich earciaoffla ; 

Prelisiinary experimeots have indicated that neither 6- 
Mercaptopurine m)r N-Hethylfonoamide have the capacity to extend 
the lifetiiae of loice with subcutaneous isxplants of this tussor. 

Signifieaace to the Program of the Hatioaal Cancer Institute ; 

With the ea^rgence of msaezous ch@&ical compounds ^ieh are 
"active** in vario\ss priissaiy screens, there has been a need for a 
p3?ogram devoted to deseribiag quantitatively, the relative anti- 
tumor specificity of drugs of interest. In the Drug Develojaient and 
Evaluation Program assay proeedtn^s for detersaining the relative 
antitumor effectiveness of drugs aore utilized estployiag prolongation 
of the lifetime of tusaar bearing M.ce as the chief criterioE of 
therapeutic effect. 

Ia addition, with the srealization that laassy fectors including 
the schedule of tx^atmsnt, the roxste of adcdnistration, az2d the xise 
of coiiibiimtions of drugs, as well as the experimental test system 
oaployed, i^iy in^uence drug effectiveness, pharEffi.cologic studies 
of the optijaal conditions for antitui^r activity are needed to 



o <-» 



Caieadai' , Year l^p9 
Page 18 

pTOvide an accurate apprasial of the i»tential useAslziesB of 
ai^ c<xspou&d. 

Thusy the Drug Developraeat and ISvaluation Program has pro- 
vided a vaewas for helpiisg in the selection of compounds of great- 
est potential intearest in clinical loveeti^tion. In the close 
collaboration between the Drug Developoeat aad Evaulation Pro- 
gram and the Clinical Prog£^a of the ISOI it provides the means 
of investigating clinical ^rdbr.'ssm at the aninial level. 



During the next year, the various aspects of this progrsa 
vlll be continued and expanded to include other test syst^se 
vhidx appear to be appropiate. In the testing of new cas&^vsn&s, 
particular saphasis vill be placed on structure-antitusaor activity 
relationships. In addition particular attention vill be given to 
factors ^ich tend to alter the host-tuiaor-drag relationship such 
as the schedule of treatisent, route of drug adniinietration; e'Ic. 
Since one of the chief ffectors limiting the therapeutic wsefijl- 
ness of active drugs is the developis^nt of resistance to ts^at- 
tsssnt, additional emphasis viU te placed on studies of drug effects 
in various drv]g resistsnt ttoior systems. 



33^ 



\ 



Serial Mo. Mei°2l6 
1. ICI 



2. OADB 

3. Bstiesda 



FHS - NH 
Individual Project Report 
Calezidar Year 1^9 



Part_B: Soaors, Awards, aad Publications 

PuBtlieations othsr thaa abstracts from tMs project: 

(l). Venditti, J.M., Suarphreys, S.R., aai Goldia, A, 
The Eff^sctive^ss of (^toxaa Agaiast Sfsuse 
Leiikessia L<>1S10 and Resistant Sttblises. Caacer 
CbeDsothers;^ Reports ^: 6-8, 1959* 

(2). Veoditti, J.M. p SiEsrphrsys, S.R., aad Goldia, A. 
Investigation of the Activity of Qytoxsa Against 
Letikemia L-ISIO in B5ice. Caaesr Research 19: 
986-995, 1959. 

(3). Goldin, A., Veaditti, J.M., Klin®, I., end 

Mantel, H. Evaluation of Aatileuksaic Agjsnts 
Employing Ad'yaaced Leuksmia in Mice. Canear 
Research, Supplement "Cancer CSiesKjtIaerapy 
Screening Data IV." Ig: i^29-^66, 1959. 

(k). Veaditti, J.M., Frei, E., Ill, and Goldin, A. 

'She Effectiveness of I^rezoloC3,U-d)pyrisd.dines 
Against Transplantable Mot^e Tuoors. In press. 
Cancer. 



335 



Sesial NOo NCI-203Ca,b.c) 

Cal«adsir Year 1959 

lo OADR 

2. Enviremrtec^lal Cancer 

3o Betheeda, Mdc 

PHS-NIH 
ludiidduai Psrojeci Report 
Calendar Year 1959 

Payt Ao 

Project Titles ^ M acgomolecalay Polymers as Cagciaogeoic A^emB 

&„ E3tpeg|gg>eatal Studiee. ia Wafeeg Soluble M&cgomolecular 
Chemicals, PolyviB.fl Pyrifelidoaeo Dextranj Polyviayl 
Aleokoip Meth yl Cellulose „ S^gch 

Priaeipal Inv^sUg&tor ^ W. Co Huepe? • 

b, Egpegimesttal Studies era Wa^eg°In8olttble Macromoleculaff 
CS&eaiicalsg ° Siliceae Rabbeg, Silieome Gmn, Silicap Poly° 
e ^yleg^e, Polytsgethaae , E th^l CeUalose 

Priacipal loi^esligator - Wo C. Hueper 

Co The Syalheais of MacgQgnolecBlag P&gaffias ^ 

P?iacipai lavesiigator = Jo Ho Seipel 

^ Discosiltiaued due to the departt^^e of the principal imres^gatos-. 

N© Pa?t Bo 

Maa Yeaga (caleadar yea? 1959^ 

Tofels <=»„ ^ »„«,.<., 7 
Professicmah - = = = «- «. 1 

Others =^.c. »»»-»„6 

a. Expegjineatai Stadlea i a WaS er Soluble M ac goimolecttlag Chem ieal s s 
Polyvinyl Pyggolidoa®, Desefegaap Polyvinyl Alcohol, Me&yl 
Cell ttlos e, Starch 

Objectivess - -- Numerous wate? soluble oatu^al aad eyathetlc polyn-ierised 

macfomolecular chemicals are extensively vieed for 
numerous industrial asid medlciE^l pus-poses., nuchas hair lacquers, 
esnulsifying agents^ protective films, food tkicke&ers„ Ia:Katives„ prctheses 
plas?na extenders, pM^nnacologie retardae>ts, delOKieat^ts » aad sus;pendii^;g 
asd seslubiliziog ageats o£ medical preparatiot2S« Since the paresitesral 
impiazstatioT^ of a eotisiderable auxnbes^ of water insoluble polytrt^rs; had 
reisulted in th« develop-ment of sarcomas at the site ©f depositloa^ it ^as 

this property asd whether any <^inc^rou« reactions resulting from their 



- Z - Serial No. NCI-203 fa) 

Calendasr Yeay 19§9 



_a -<co5&t) 



i^tr&ducti^a iat® 3Kp@riinea%al aaimals migM depestd upoa aay specific 
chemic&l cosnpcsitioa or moiecula? structural peculiarity f shape aud 
ffiise «>£ molecules cross licking, molecular malformations „ mature of 

re3.cliTe esd gy©up£|= 

Metl5i2^3 Emplofeds ■= - The lon©-«?iag water soluble carbon polfmes-s 

we^e us ad ia tMs study? - Tea PO^ dilfeffaafc 
types ©f polfviayl pyrrolidoae of A!mesficaa„ GermaR and English inaiiu<- 
facluirso rass^is^ ia average moleculai- weigM between 10,000 a«&d 300,000 
aad varying ia the cSjaractex- of ^eir e®d gro«p8„ cross liokageSo aad 
mal£os'Bniatic»t£3 b€»cau@« of diffesrei^t catalysts employed is thei? maaufacto.v'e 
Tlie pyedomiaasal xtiolecisis? shape nsve3Ptheles6„ fo^ all PVPa used ia a 
straight c&aia« Thsf®® ©f the PVPs weg-e ®f the tnedicioal type, the others 
wes'e eiti^es' lechmcal pr€»ducts or specially prepared samples having a 
Emy^ow molec^ala.r g-asge asro^Qd 20e000» 

Eleven diffe.reM types o£ dext^aas were testedo These polyglucoaes 
varied uot ©sly greatly in nnclecula? weight (from E4„ 000 to several milU®!3il 
but also ill molecula£> shape^ io eo a l-tmti straight chain molecules to highly 
hs'anched molecules „ 

Polyviayl alcohol aad methyl cellulose of medium average molecular • 
weight farouad 60,000) and a soluble starch of unkaovm moleciiiar weight 
t(;e%°e also tested so as lo e^lead the spectrum of these chemicals studied 
for obtainie^ the basis fos* a more ccmprehemsive evaluation of pelfviayl 
aad dextrose polymers = 

Mice |C5? MackK r&ts CBethesda black) asd rabbits (Dutch) ^ere 
usedo The polymers always severe implanted as powders into the sub^^ 
cuta^aeous tissue aotd into the peritoaneal cavity of inice aisd rats aad 
were i&Jected as an aqueous soIuti@i& iato ^^ vein and peri^sseal cavity 
of rats and mice in coai&ecti<»8% with the testies^g of several PVPs ai&d 
dexlraae. While isi most ie^stai&ces where the polymiere were implanted 
as powders a sii^le admisustratioa was used, in several ejqjerimenlo of 
this type repealed implantatiens at the same site were made. Solutions 
of the polysners always were iajected repeatedly. The amounts used were 
ia part coasiderableo i» a«% attem^pt to produce storage pheaonneaae 
eimiiar to those reported pre^ously in e:£persmeats with polyvinyl alcoholr, 
methyl cellulose, arad pecti!i„ siace as&y taeoplastlc reactions which might 
be elicited might reflect the local aad systemic distributioa aE);d reteatioa 
pattern of these chemicals in the bodyo 

The obs ervatiGiRia made in the present studies ixk regard to carciiao^ 
genie properties of VVP® cossfirmed those previously made ia a gJDiallsr 
acslelCa-SiCSi'c 10^8- iS„ 195'/^ Tijey witteadad them to some of fht followiBg 
water soluble polymers iDextraas, PYA, Starch, Methyl CeUul«*seK The 



C&i'3ii«Jiatf Year i9: 



lollowisg coaclusions were reached ivom si.n o^^alua^oa of the re suites ei 
the Tas-ious fioisfeed experimeoSs? 

1» Ce?tait% water- soluble polysners, auch as polyviaylpy rrelidoaeo pelfvmr- 

alcofaolo &ad deKtraa elicit upom pareKte^al ii&troductioK i&to r&ts end 
mice„ and perhaps al8@ ia vabbile, sarcoznae originating from orgaas and 
tissues in which these macromolecular substaaces are retained and steered, 
io Co „ the reticuloendothelial %is9Uea. Casucers from other tissues, such as 
uterue as^d skiss, observed in rats may b® directly or indirectly caused h^' 
these su staaceco 

2o Polymers &f the same general type but produced by differeat procedurc^r? 

as&d varying axnoag each ofher in molecular -weight aad configuration 
differ greatly in their carcinogenic potency, ic eo , some are noncar«:iiS£»geMr2 ..- 
others weakly earcinogeniCo and again o^ers definitely carcincgeaiCo 

3o The experiznents conducted have failed so far to provide any informacior^ 

on the physical or chemical molecular factors which jm&y acc©UBt for 
these differences 

4o Experiments wi^ silicon rubber and silicon lates: shds^ed that the srubb«:v 

havimg a net molecule with many cross linkages possesses marked 
carcinogenic properties when eubculaneously impla^ed iiia rats^ -vyMl!? ^^'-'^ 
latest having a linear molecule is at best weakly carcinoges^c. 

S. The l@tal evidence tm hand concerning the biologic action of synthase 
and senni^eynlhelic polymers parenterally iis$:roduced into ms.f% a^d 
animals favors the view that their tosie, pharnnacolegic, and carciffioge&ic 
effects are mainly related to chemical properties and reactions r^thesr 
than to their physical characleristicSo 

60 The existence ©f actual cancer hazards to man frorn larater- soluble 

and ii&soluble polymers possibly associated not only mth their 
parenteral introduction but also ^ith their admioistratioa by the cutaneous-, 
respiratory „ and oral routes ie at present problematical.. The expevimmrJ-Bl 
evidence on hand, h©wev€r„ is sufficiently definite and serious for reqins-ing 
distinct caution in the medicinal and cosm^ie use of ^ese prs-ductSo 

Tentative observatieins only can be presented at this time of the other 
experiments, which have been terminated during 1959 because the 
hie^logic study has partially been c&nnpleted (table l|. 



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Gaieadar Yea? 19S9 

Sigidficsaces - » Tise additiosxal obeasripatious on PVPs fs-om wMcfe Mgh 
' molecvul&r conslituettts have beess, removed aisd which 
^sre admimsterecl ia physiologic salisae iMo the pQ£>i$oQe&l cavity of f&ts 
and s>ftbbite iudicaSs thai such PVPs lack under: ^e specific eacpearimeatal 
conditions, caeicerigetiie p?opertie@o This pheiKomeaoa maf , ia part, be 
df3@ lo the fact that their administration in a dissolved form provided fe-r 
a ma^ksd asad sig^ilicaat dilu^©^ of these cheBnieals al asi]^ ona pa.^t (d 
the host os'gaoisnie in p&^t it nri3.y be due t@ Iha fact that the ^elal^vel^ 
lo^ average molecular "weights &f the PVPs ussied iu these experimeats 
facilitated their rapid excret&ea ia the urine &ud Iherel^^f counteracted 
thei? proloiaged retenUosi arad storage ii% ^he hodfo 

The experiments on this project have been ^ermiaatedo During 
the coming fear the histologic studf will be c<simpleted aad the results 
^ill be evaluated and reported isi. a preses^atiost before the American 
Issdustrial Hygieae Associatiogs iu April, I960, together vyith fiadings 
obtained m iavestigatios&s oa polfvtreth&na. polfsilicoae as^d p^lf ethflenao 



Part B lacluded g YES X NO 



3ill 



-5» Serial Koo NCI-E03|a & fo.| 

Caleaday Yeaa? 19S9 



PART B 



Pu6>Uc&li©a®i =■ - Hueper„ Wo C, „ Experimental Carciaegeaic Sfe*^©s 

in Mac?eirsolecular ChsmicalSo He CareiKSOgemc 
Studies OR. Wate? Soluble Asid Insoluble 

Macromolecttla®. Arch. PathOo 67s589»6J'?,, 



342 



Ca3L©adar Year 1959 

h. E-xp&ximenteH Studies oa Wates°I®so5uMe Macaromolacuiar Chemicals; 
Silicone Rubber, SilieoBe Cmn^ Silica„ PoiyethyleE^eo Polyure'diaae^ 
Eihfi Cellulose 

Objectives? - The rapidly iacreaeiag us® aad diversity of polymer plasties 

in is^dustry, Aeir gtrowiss^ efnpiofzneat i» coisstsunner goods 
aad thei? parentes'al iiat2roductiai& iMa tha Imxnaa body £&k nrisdicioal purpi^^se..' 
laave c^reatcd a sezioias problem from a public heaS-feh vie^wpointo siaee maay 
of file water insoluble carbon and silieoa polf^ners have been found to elicit 
cancerous reactions when ixnplaated is^o t&ts &ad irnice. Sissee maay of th&^e 
m&teriale are beiisg looked upon as cheznicalif rather isiert, ikes question. 
h&8 arisen whether the carcinoget!ii.e lesioras develop itt response to chesmicals 
released slo^lf from such polymers under the isiflueaco ol metabolic 
processes of ^he host orgaaiam. or whether they depeiid upoa surface forces; 
related to the physical state of a polymer » u eo „ whether it is prese&t as a 
€ub@o as &n ivkt&ct or perforated film or ae a ponder , a-nd whether it giv^s 
rise to Ihe formati€:«% of a fibrous capstsle -t^^hich allegedly is esseiatial i^s 
the subsequent developmera^ of a sarcomatous growth.. CoraeideratloQ, snore" 
over, had l@ be givea in ^is respect to the unsaturated szature of the end 
groups of these polfi:ne7s aatd to the possibility that in certain polysners 
side cimiras might undergo chemical chaages in the body^ io e, „ in poly- 
uretha&« polyraers the cfaao groups, for i»stae&ce, migh^ be reduced to 
amino greups aad the moaomer msmber of ^e polfmer maf assume a 
elo@e reaembiasjice So certeia carcissogstsic dipfeea^l- aad tripherayl metJaaee 
df eso These coaeideratioias entered i«%$© the plamm&g of the ezperimes^s 
oa water i^^eoluble polfmerso which coxnplsmef&t those couducted on the 
water soluble macromolecules., 

Methods Employed? - - The p^lfmers available as filmEt, plates, rods 

sponges or powders were implansted subcutaoet'f.;:.. 
aad/or iatrap^ri^aeally iat© mice and/®r rats (C 57 black mice, BsUthestSa 
black ratsK Fiftf (§0) mice aad thirty C30J S"ats were ususllf empl©fsd xc-s 
each polfmer, each physical istat©, aad routSo The followiag materials 
were thus testeds -- Polfeilicoae rubber CSilasticK polfsilicoae gum, silica 
add ©d to the gum ia. the preparatioa of the rubber similar t» the addltlcm 
of carboa black to the aatus-al aad syot&etic carboa gumsK polfurethaae 
plastic available as a sheet, sponge, and powder, ethyl cellulose povrfdes?, 
and polyethylene film, cube, and powder (Table 1|= 

Major Findiugss - =■ Silicoa rubber having a liel molecule resultissg from (;>;<- 

vulca®izi!^ acfeioa ol benzol pero-xide upoa the late:, 
displayed a m®derale carciBogeiaic acfeic.® upoa the cenaective tiesu© aui . <■.-.-.- 
ii%g eubcuta&eous implaiatSa Silicoee gum seemed Io be ii^ert in this respisct 
al^ough being appareatly qui$e carciaogsBically poteat when iKlraperifeaeaU-^ 
iatrodueed.. Powdered silica is^as carciaegeaically iaert ^hen subcut3VB.eou&I:|; 
or iatraperitof^eally adznimsteredo 



Caietsd&r Year 



h Cecal)) 



Poiyethfieae eshibited £oliov/iffig i*a iafarapej-'itoaeal lEwpiaatstioa a 
moderate carciaogetaic activity jregapdlsss ®£ wfeetSjer preseall aa a eube, 
a disc o? a filmi» It was apparsatlf iaactive ^laea iaSroduced as a po-sfdefft 
although small punctiform white deposits or plaques where aot iraf^eqiseatly 
noted oa the seF&s&l surface of the spleen anid livero 

PolfUffethame plastic implaated subcutaaasmsly asad iat?aperit©ReaUw 
appeared to exesrt a carcifiogenic effect oa the tissue at the site &£ irfl= 
plaatatiea wham i{^?@duced stubcfstane&usif as a spoQge. 

Significances ■= ~ The ©bssi-vatioae made with fthe three wate? iaaoluble 

polfmes's admialstered in vairieuo physical forma de sot 
8uppo?t th© view that surface forces play the pyed»mi!^Et role in. determimag 
eftnceireus s-eactione. The fiodisEgs and coRclusieiiMS „ theyefore. ref«te allega- 
tioas of Oppanheimer et aL aisd ©f Nothdvis-fto ^ho assert that the sub- 
cutaneouB p©lymeir tsamo?® are essasa^aily sarcomatous foreign body 
reacti@raso 

Future Cowf «©s = » Als© this phase ©f She present macs-omolecalar s^disE 

has feeea lesmiiaaled i&^ the Same feeisjg because of 
iraadequacf of space aud pe?s©s&Biel 'fvMch had t@ be used for projects ai 
snore urgeisl i£npor(^E£ee« 



Part B Included? YES X ® NO 



See page 5 NCI"203Ca &b| 



3Uia 



Caieadaj? Year 19'- . 

I. Office Assoe- Dis-eetfflr 5a 

CSsag-ge ©£ Rassas.*clj. 
E. Ef8«vi?o!snoes5oal Caacer 
3o Bethesda 

ladlvidttal Py©jec4 Report 
C&leaday Yea? 1959 

Part A» Project Titles 

GarciEtogeKiic Studies on Petg^alftam Sabstifattes aod S^pt hetic 
Liquid Fueiss 

a, Ca?cin©gffiaicity of Shale Oil Fracfi©ns of L^J-w BolUsig Poisat . 
astd F?ee of 3,4-BeQspy7e!:^ 
TERMINATED 

bo Carcisi©g€Bi.c Stjadiea oa 3o4-Benafluos'aat&eae 

TERMINATED 

Cc Ca^cieogeoic Studies ©la Road- amd Roofiiag Agphalt^CPets'eleaizi) 

Priacipai IsvesfeLgator = W, C. Haeps^r 
Otheg- Investigates - » = Wsn. W. Payne 

Mas Years fcalendar yea? 19591 

Totals -= „™--- „? 

Pfeles sionals •= » - ~ - - - - » , 1 
Other? »----- — «.^» — - ,6 

c, Carciaogestic Stadies on Road-> aad RoofiRgAgphaiSslpaSgetlevuTi) 

Objectives? •= =• The ejtteasi^e usis of petrel©iS5ri asphalfes f&? Bazjnsreiai? 

iaduaferial purpoecs ©sstailieg cutaaecus, respi^atorf and 
irageativia coistact of large aiunbers of ioidividual-s v?ith these agecets fo? 
occupational asd enviffssjmealai ^easoas uscessitated aia esplas-siSss'f study 
of ome Tooiing aephal$ &vs!d four differesi^ road asphalts ior caircisiegesic 
pyeperlies sissce reliable data ©a Shis aspect seesned to fee lacking is ib.e 
published lltes-ature, although available in She records &£ asphalt matau-^ 
facterearso The asphalts used in ^ese experisneats were v^t biesaded -with 
&nj fuel oil &s solvent.. Thd fuel oils used for the purpose of curisj^ ?oad 
asphalts are likelf^ to possess carcis>@geaic properties. The road s.ephalts 
studied were supplied by the Bureau ©f Road Ccastrueti©a„ Uo So Dsp-arj.-* 
meat ©f ^e foterioro 

3ii5 



serial No. NCX»a04Cc) 
Caleadar Year 1959 



c. 



Metheds Employed? =■ - Mice wers p&iated 2 tisnes weekly vfith the 

asphalts as well as ©ace intFamuecularly iojecited 
with 0„ I cCo of asphalt im tricapryliu.. Fifty C5? black zraice we^e employe 
fd? each asphalt and fosr each route @f admissiet^atioiio Thirty xraftS^Be'^hasd 
blackj vtete isslramuEcularly issjeefted with 0, E cCo e>£ asphalt. 

Fi^p& (B) rabbits V9&sq palnlsd on the dorsal surfaces of both ea?8 
as&d on two aitas of the back vdtSx each of the four road &aph&ltM twiee 
a weeko 

Major FinditEgss °^ <» Histologic evidence of cancerous ?eac^ous at the 
sites of applicatiof!^ of the asphalts >sras obsesni-ed 
ia znice and rats aftes* Z years of treatment, as sumnia?ised in tha 
follo^^ing tables C^aMe I „ page 2al) 



Sigmficasaces - - The obsarratleffis made indicats that road asphalts 

poaaeas a relatively weak carciaogemc property f.QZ 
the epideriTiis and connective tissue of znice as&d ra^s. Th@sa findings 
affe ©f aeroi® inripertaacQ froiyj the vie^'poiat ©f possible sources ©r urban 
air polluMoa aisd of occupafloaal cancer hasardso 

Proposed Courses ^ - Aitar a hisfol@gic study of the pertiaent tissues 

of &e ammals used has been made^ a bffief tsp&T'i 
will be prepa^'ed fof publicatioi^^ Fur'&er escperim eettj ok ?Ms-mates.'iai 
are not ce^teniplated at the present timeo 



PART B Included? » YSS X 



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Caleadar Year 1959 



PART Bs 

PubllcafcioEss - - 

Hueper, W, C. and Payae, Wkio W= » CsrciaiBgemc Studies ©a 
Petroleum Aaplsalto Cooling Oil and Coal Tar. 

Accepted for publicatioa ia the AMA Archives &£ Pathology. 



3il8 



L OADR 

2, Enviroftimeata). Caacer 

3. 



PHS-NIH 

Ifidividual Project Report 
Caleudar Year 1959 



Project Title;- E^pe Hm eatal Studie s^ in Met&I C ance gjgeaesi s 

a, Experixneffital Studies on Caaceffigeisdc Propes-ties of 

MetelUc Nickei 

i= P?©loffiged ia halation of Povydered Metallic Nickel- 

ia Cotnbiisatiea wiih Sniall Coaceatratioa of Sulfur 
Dioxide by Guinea Pigs, Rata aad Hairvsters. 

Principal Isvestigator - W. .C- Haepes 
Othey lavestigatoF - « - Wrsi. Wo Payae 

2» latrapulsxioaasry Sajecti®a of Nickel Suspeasicmg ia 
Gelatiia Solution into Rats. 

Priacipal Ittvestigatos' - - "W C. Hueps? 

b. Biologic and Biochemical Studies ca the CaBcerigenic AcSdoiu 

Chroniium aad Chxomixxsn Coirspounds 

lo latrapleuaral, iQlramuscQia? aud £Msrat?acheal Injectioas 
of CharoiBite Ore Roast aad of Several Pure ChrotnimTa 
Compousids 

Principal Xaveetigator - - W. C. Huepsr 
Other lavestigato? =.--».■»- Wxrn W. Payee 

2, daxofflic Iiihalatioa of a Mi3£6u?e of Chroj^ie® Os's aitid 
Chromite Ore Roast with L«imeatoae Duist aad its 
Effect oa the Respiratory Orgaas of Mice^ Rats, 
Guinea Pigs a^td Rabbits. 

Principal lavestigato? » - W, C. Hueper 
Other Investigator = = __«* Wirj- W. Pa'/tse 



349 



3o Claarftc :,>n aad Bi©asi&-&¥' ^ Dust isom a 

Psitecipal XaT(88ligate^ = Wir.„ W. Payae 

OlSier laveaaigatOE- » - » W, C„ Hueper (Pathology), 

4. Biochemical Studies ia Cteomium Caaceffigeneais ■* 

PyjQcipai Inveatigatog' - - C. H. Grogaa 
OSfeeg iBvestigato? = - - =■ W. G. Huepeff 

THIS PROJECT HAS BEEN TERMINATED CB='4K ^ 

So P^olossgsd loluilaMoii of Chrosnie Oxide! and of Ziac 

Chromale, Bas'iuzn Chroma ta aad Lead Ch£>omate 

fey Ratjs. 

Priacipai Investigator - - W. C Hueper 

Other iBvesfigatoff - - = - Wm. W Pay ae 

Man Years Ccaiaaday year 1 9591 
Totals 2, 14 

Professiotmis „ 07 
Others 2„ 07 

a-L Prolonged ijshala^oia of Poswdejred Meiallie Nickel io Combiaatioa 
with Small Amounts of Sulfuj? Dioxide hj Guiaea Pigs, 
Eats and Hamsters., 

Objectives; => «■ Epidemiologic data oa caaces" of the nasal cavitjp, aaaal 

si^ue asjid luQg m mckel matte refiii^es-y workers it% Eogla^d. 
Canada aad Norway indicates the esieteace of a a occupational respiratory 
cancer ha2;ard fo? members of this worker group,. Similar investigations 
oa othe? worker groups i^viag contact larith dust, fmnes assd xBists of 
nickel as&d its compounds are oone^sistento Sifice '^os'kers i^ such operations^ 
have coatacl -with chemicals other than sickelo it was impertaat ta demaa* 
strate by experimeeital znethods the exact chemical nature of the cairciuoge 
ageat to which the particular workers become snpoaed and "^hich elicits 
the caneers in different parts of the respiratory tract. 

Doll. R. , Cancer of the Lung aisd Nose in Nickel W urkerso Brit. J. 
Indust„ Med. 15s 217. 1958. 

Morgan. !„ Go « Some observations on the Incidence of Respiratory Casaces"- 
im Nickel Workers. Brit. J. Indust. Med. 15s2E4. 195S. 

V/illiams„ W. J, , The Pathology of the Lungs in Five Nickel Workers. 
Ibid, iSs23S. 1958. 

Suadermann, F. Wm. and Kistcaid, J„ F. , Nickel Poisoning, H. Studies 
on. Patients Suffering from Acute Exposure to Vapors of 
Nickel CarbonyL S.A-,rU-„rA. 155§889o 1954. 



SuadermanB, F. Wm, et alo , Nickel Poisaaioig, IV» Chyoaic Exposure 
of Rat® to Nickel CarbooyL Arch, laduatro Healtk. 16; 480. 19"'' 

Suadermaan, F. Wsm, el aL , Nick«l Poisoaiog, .IX„ Carcii80geiaeay.is .;.; 
Rata Estpoeed le Nickel Carboaylo AMA Apch. Xado Health. 
20s 36-41, 1959. 

Methods Employed; - - Pgevioua expsrianeats had demonaisrated that 

metallie nickel pareitttesally iufiroduced i&to ihe 
feznux, muscle tissue and pleural caviiy oi rats aad/or rabbits causes 
the developmeat of sarcomas ie» aa appreciable number of such aedlals. 
In more recent expeyimeats on guiaea pigs arad rata esposed for up *o 
two yeaya to the iahalatioa of swetallic Rickel duEt, i% ^&s showsi tsvui 
practically all guinea pigs and snaay rats developed rnult-icaotric adess.©- 
iTialoid formatiotis of the alveolar liniog aad hyperplastic p^oliferationa 
of the termiaal broachiolar epitheliui«o One guinea pig had an anapla.i;^:,le 
carcinoma of multice&tljric origin affecUiOig th@ lung. 

Wi^h tMs conclusive evidence oa the carcii^ogeoic pB'opeg'tias c^ 
aickel available, it becaixie pertiiseat to coaduct inhalation espe^imepts 
wMch duplicated more closely e:sposu3re conditions eMstimg to mdu^^tri&I 
operations with dexiionsHrated yespisratOffy cancer haaards to moi'kev&. 
Recent investigationa osa pulmoaary cancer haaards from general uarbar? 
air pollutauts and fwQsn ciga?e^e smokiEig had called at%e;%Uo9. to the 
probable potentiatibg asid suppoFtiag action ejs.evted oz^ ^e sffect bf 
respi3Patory .carcinogens by noaapecific respisratoiry irritasts, such as 
oxides of nitrogen and sulfus-^ Siace sulfusic acid is used in the s-sficdng 
of nickel ore, workers employed ia this operation have contact ^>9ith 
sulfuric acid znist in addition lo becomiiig e::^posed to vafio^s type0 of. 
nickel duet, fumes and vapors. 

Rats and hasmste^a a.se being exposed to metallic saickel du0>; ccv..- 
bined v/iih sulfur dioxide in very low conce£^t?&tiota@ , so as to teet ths 
effectiveness of this mixture in carciaogeaic reapects ©^ iche respiratory 
system^ Guinea pigs oyigiaally cRlered in this eKpsjciment had to fee 
withdrawn because of an extremely high mortality fyom subacute 
pulinonary inflammatioffi. 

Major Findiagfi.;: - ~ None ®o far. 

Sifoificaaces ♦ The experiineBts may give valuable iafo?matioa os tba 

practical ixnpoietance of missed ezposures (carcisogen 
plus irritant) frequently encountered under occupational as vs^ell as g 
industrial conditions in the production of cancers of the luag,. 

Proposed Courses = The experiments will be continued until a masdsT" ! 

exposure period of two year® has fasea reached, 
posure will be completed in about 12 moathSo The a^imr?": • "■■'' he ohfiK-.iv 
after exposure is completed until they d5~«i1 
PART B INCLUDED Yes X ^-'•' NO 



Serial No, NCi205(alJ 
Calendar Year 19S9 



Publications; =- Huspsr, W. C« , EsperisyieBtal Studies? Ib Meiai 
CaacerigeaeaiSo iXo Pulmonary Leeiono in 
Guiasa Pigs aad Rats Expoised to P?oloiiged 
lolsalatioa of Powdered Metallic Nickeo 
Arch. PathoL 65;600~607<, 1958. 



352 



Caleaday Year 19§9 

&~Z, Intra pulmooary lojeetion of Nickel Suspeasioos ia Gelatin 
Solutioa Into Rats. 

Objectives' - ■» Previous experiments asiog a pultnoaary itats'O^Suctlon of 

meteliic nickel dv^st by iahalatio» had givea a relatively 
nneagre yield of preneoplastic and neoplastic lesioaso Because of the 
possibility that these results might be due to eithe? an ial7apulmo2&&ry 
introduction of insufficieg^ atnouiite of siickel or to s.n, inad®qu&te contact 
of the iislmled nickel with the bronchial mucosa, if was considered 
desirable to inject definite axnounts of po>i?dered nickel directly into the 
lunge of Bethesda black rats in an exploratory experiment for testing 
the validity of these assumptions. 

Methods Employed^ » - Approximately 4 mg. of powdered metallic 
nickel in a 10% gelatin solution was injected 
through the thoracic wail directly into ^e right lung of 34 female 
Bethesda black rats, about 3 months oldo 

Major Findiages - - Noae so far. 

Signifiea&ce; » -The growing interest in QOQ=>tobacco agents ^rhich, as 

occupational and envi?oame8,tal air pollutaii^te might 
take part in the production of cas&cers of the human iu^„ make advisable 
a thorough investigation of various metallic ages&ts £g« carcinogenic 
properties I© the tissues of the respiratory tract of man and e:^perim9iital 



Proposed Courses • The rats will be kept under observation for a total 
period of 2 years. 



PART B Included- ---»'" Yea iX) ; NO 



353 



Sejfial Ho. KCI20§^a2) 
Calendar Year 19S9 



PART B 



Publicatioas; ° - Hueper, Wo C. , Expesitn eatsl and Hietologieal Studies 

of Metal Cagscera ©f t,he Lu^o Abatr. 7th 
lateraato Caacer Coagrees, JLoodca, July 19§{ 
Publo " ACTA Uedoa latarsatioiiaie contre le 
cancero XV|2);424=436„ 1959. 



35H 



Cb^oznite Ore Roast asd ©f Seve^sil Puse ChTe.mi«sn 
Co3np#%&&dSo 

ObJeeHvees <^ ° Despite tiie d«mee%et7atios e£ & ses'leus lu^ essiCdS' ha^^erd 

States aad Grsat BfitaiQo 3ll attempts snade by sever&l iavesMgatore to 
pr&daee such cancers isi aBinnals ey the use of va.rioua es:p@rimeislal pso- 
ceduFds atud ckrczni\2sn compounds had failed ot givees equsYi^cal results^ 
Sia;ce i^ appeared possible ^hat these failures might be attributable to the 
fact that substances we?e used ^ivhich either tve^e itot aufficieatlir soluble 
ia a Dfatery physiolosic mediuzxi for percnittiag a release of adequate 
annouQts of biologically active chromiuzn |chroraiium snetal, chremite 
ore^ or '^ere too corrosive and therefore e^ertisig aa. exclusively 
Qec?oliai@g effect upon the cells upon coaiact, chromite &ve roast was 
tested because it coQtaiis&s ia addition to sodiuin nicisochroniate and 
chromite ore, various intermediary stages of osidized chromium c^n^ 
pounds as well as chromium polymers being of moderate solubility and 
lacking acute neerotisieg qualitieso 

The apparent validity of these consepIS «^as attested to by some 
prelisTiinary experiznsnt&l results obtained iu rats ^Mch received intra° 
pleural and intramuscular implaists of c&romite ere roast suspeoided ia 
sheep fato This ^siaterial contais&s various chromium com pounds differ^ 
ing markedly in their selubilily ia water o 

Three out of 31 iiatramuseularly implanted rats developed apiadle 
ceil sarcomas- while 2 out of 2S iatrapleuraliy irnplai^ed rats developed 
squamous and adenocareiitomas of the iuags, complicated is one rat 
by sarcomatous lesions. 

Methods Employed! •= - Chromite or© roas* produced and used ia chros«aie 

plants is either ob^izied by ^s rcasMng of 
chromite ere only ^primary roast) or represents the product of roasMag 
a miscture ©f ehremige or© and the leached residue of a previous roasfe 
^secondary roaetK Siace„ is th« o^igiEial experimsB*, & misfeir* <^ fee 
t^o roasts ^aa used, it vi&@ important to detersiiee "whether ^e tvi& types 
of roast might vary in carcinogenic potency, because they could be 
expected I© differ in the quanlita^ve and qualitative compcsitioi^ of 
chroxnium ceixipoundSo T>ivo sets of 35 rats each received implants ©f 
2S mgo of the primary roast residue Ixnost of the sodium chromate 
removed from roast by leaching at the plasit) into the pleural cavity and 
int© the mus;cle tissue of the right thigh, respectively, Many cf the 
animals are still alive af^er 2 yearso 

Because o£ the partly uncertain and varying nature of the chromium 
compounds in chr©mite ©re roast residue, additional experiments «?e?e 

355 



Cait&dai? Year 1959 

Metbode Employed Ccoat^s 

set up ueisg pure fabovs 99%^ chi'enfsiurn connpounds <d£ffe?iag among each 
other io. thei? relative soiubiiitf ii^ v/at@?c sosne of which po«aibl|r ievmed 

duFiag the 3foa@tiizg p£>oees9„ e^ers eKt;ei&sively emplaysd m. the human 
ecoaomy as pigsneMs assd aaticosreosive® Ic&lcium <:^^©mat«c siatered 
calciiam chrorsiatis, magmesiusri ch^oznaSe,, siatered znagnasium cSiromatee 
Btrontiwn chsosns.te^ siiac chyomate, barium chrosnateo lead chromate 
aad aiates-sd clssmniun'ii teiosdd© os chToraicchxoxnaLteiCx ^0^2)1' -As in 
ps&vimiB e:speri^n@atoS„ impia&ts co^staiaisig 2S nngc asad suspended itt sheep 
fat, V79i*e iEtrcduced isxte^ ^e muscle liusue of ^e femur aed ioto the 
pl®u»al cs'^'itf „ refapecSiveljTo of sets of 35 s-afes each. These experimealal 
appg'oachee have s'essatLf beets suppiemeslQd by the repeated monthly 
iati'stts'acbeai iisjectioB. ef Oo OS cc. of a 20% solution of gelatin c&staimiQtg 
about Z zngo of the parUculas' chr@Dnium eompouud ^calcium chroxnate).. 

Th© offigiaal ©jspesrimeaial approaches have baea supplemei%ted bf? 

L I^j-teapl^ural aod iMraxnuecula:? implantation of chromic 
a<e state ia gelaMsa capsules ix% rats. 

2o lijitratraeheal ads^inistration of calcium chromale in 

gelatisi siolutloa t® ?&ts. 

3o lalrapieusral aad iiateasnusculsj? iajeclions of sedium di- 

chsqmats solutioia to rats at snon^ily iatervalso 

4o Satifats'acheal admiaistrafien of stroiitiu»n chromate its 
gela^ia eolutiei^ te ratso 

These supplements to tlie oirigiffial esperimej^ wefe uadertakeig&t& 
t^y te produce casrcisaoma c€ the luag, te ascertain ^whether trivalenfe 
chromium compoua<3@ will also produce tumors ii^ asin^als aad t& deler^ . 
miote whethes* the re^m&ted adsniedstratiosi of a highly soluble chrosnium 
compound -will p;?odac@ the sasx^e results as a depot of slightly soluble 
com pounds. 

Majos" Firadingss =■ - Both the magiaesium chromate aad the siate^ed 

magnesium chx^omate wese too toxic to adi^ioister 
it& the same do9&g@ a^d mans.er as ^e other compouadso The aumber of 
tusTiors produced hf the various chromium compouods and the solubilifc|' 
of t:he substances are summarised belowo 



356 



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357 



- iO -' Serial N®c Na^ZOSP?!) 

Cal@i!s4%r Year 19§9 

The c&ne®Te of the thigh wesre apiauile and fibs^aarceacna, those of 
the niediaetiisuxn i%v@Ivitig o£t«iE!i tSie hea?!, luags aiui Ifmph nodes, were 

ueu&Uf of the sasne bisldlogic tfpei ojidy Qceasiot&allf thef wes'e primarf 
in the lusig aad @f squaznoue cell tf pe^ 

Sigoificatacss ^ The observations establish the fact thai chrmnium when 

p?ea«nt in prosper biologically available form» is car- 
ciiaogeme to rats and capable ©f eliciting carciK^oomas of sqvtaxnous cell 
&m6 adenoeaircineinatoas t^ pess %!& the lungSo These findings SB@t onlf 
p?<sve the ecc^paiional ssature of the lung cancers seen in chr@xnate 
ma&ulactu£'@£'s b^t provide indu«t?y with iinp@rtant information where 
heet to apply preventive teehsKsilogic inethodSo The present experiments 
field also definite is^formatios^ on the carcinogenicity of Binc<=°, barinzn«„ 
and lead chr@mates« Experimesatal etudids with these pigments are of 
distinct ps'actical Impowtanc^ since thef are used in anticorrosive prir^ier 
paimts on airplane Se sMpi^o oil lines, bridges „ railroad cars, etCo asd 
represent potential lung €ar»cer has^as^ds for Ihouisand® of spray painters, 
<welders, scrap m«tal ctstt@r& and smelter workers. 

Proposed Cou,rses <^ ° The expei'iments ^U be continued for another 
two years @o as t@ allow a final assessment of 

the result® ©btained in the individual experiments, some of ssrhich were 
started ©nly a few months agOc Since ^e main goal, m this investigation 
is almost reached , &o further experiments are plaimed at the present 
tim®o 



Part B Included- -» ~ — YES- =^^- NO-^--- 



358 



= iS - 



SesrIaS No. NCI-^SOSCbi^ 
Calendar Ye&r i9S9 



PART Bg 



Pubiieationss = Huepes, W„ C. , aad Payee,, Wnn. W„ , Experimeatai 
Caseers i» Ratu by ChposBiiun Compouads atsad 
Thei7 SignificaiBce to Xndastry antd PuMic Health. 

Jouro. of Indo Hyg= Asesa, 20s274-280jAug. L i9§9o 

Huepes-o W. Go aad Payme, Wm, W„ „ Experimental 

Caskcerffi Px'oduced ia Rats bf Chromium CompoundSo 
PuMic Heal^ Reports, Its 9iO»9H„ <Oct, K 1959., 



353 



CaJesdar Year 1959 

h^Zc Chrooic lBlt&lalio:& of a Mixture of Chircrmits Ore Ic Limeslo&e 
Duat on the Respiratory Organs of Mice, Rats, Guinea Pigs 
and Rabbits o 

Objeetivess - - Identical with Serial No. NCI-20SCblK 

Methods Employed; » <^ A prolonged inhalatioii of a miicture of powdered 

chr@tnile ore aod limeslooe hf nnlce, rate, gui&ea 
pigs and rabbits did mat reetsit i«i the. developmet&l @f eaacerc of the lung 

in these ammalSc as soled iis. the aes^ual proj«ct reports @£ 1957 aigd i9§8„ 
These negative results obtained by respiratory exposure to chromite ore 
eonfirnned thg>se previously observed following its intramuscular and 
intrapleural introducMon into rats. The positive results recently obtained 
^th an intramuecidar and intrapleural ii!<,froduction of chromite @re roast 
suggested that the chromium in tMe partly oMdised material was in a 
biologically available aad carcin@genically effective form^ Chromite ore 
roasts of the primary and of She secondary types wer# obtained from the 
Mutual Chemical Company a&d &rm used separately for reasons etated In 
project reptsrt noo NCI=20§^b°^l) in twe chambers on Betheeda black rate. 
There are lEO rata in each chamberc 

Major Fisidingss °^ -^ The fei^; ammale thait eurvini'ed the two°^ysar exposure 

period will be observed until they die^ No tumor » 
associated "ovith the exposure have been observed, but final results will 
not be available un^l hist&logic&l examina^on of tissues is c^mpleted^ 

Significances ° "■ The experiment repr<sse«l8 an attempt le demonstrate that 
chr@mite ore roast when inhaled elicits carcinomas of the 
lungs @f rats and thus has an effect identical with that exerted when this 
material is brought into direct contact with the lungs by intrapleural intro^ 
duction^ A positive outcome would supply additional evideBice it&diea^eg 
that an exposure to chromite «re roast in industry is one of the major 
sources @f lung cancer hazards in chr^nale producing plants » 

Proposed Course; ^ - When the last of the animals die„ Mstological 

examination of the tissues <^11 be completed and 
the results ^U be evaluated. 



Part B Included YES NO 



360 



- 13 = Serial Nc. NCX=205Cb3) 

Caleadar Year 19S9 

b»3o CharaetefisBAtiogi aad Bictacsay of Oust from a Chromate 
Producing Plant. 

Objeetivess ■- ^ Whsa ^bie p?ejeet was slax-ted^ a lugh iacidectce of eancor 
of the lu^g had b«en observed ainong wosrkerii ia chroma^e 
producmg plag^ts^ but Ibe specific agesit respoasible fas' this higb ineideece 
had nol been ide&tifiedo 

Methods Empiof eds ° =Iq the initial phaee of Ibe work, two maieHalss 

pulverijced roast matrix and pulverised fis-st^stage 
residue, were f;ac^ioaa$ed hf particle size and tbeir compositioei and 
properties studied. 

Tbree fractioos of ibo reaidae and a pure cbromium compoandCcbr@mic 
phospfeaiel were 8elec^>ed f@ff bio^-aseaf usifkg appreximately 600 CS7 
black mice. The fraetio3%s were cbo@e(& I© give a comparison betweeis vary ^ 
ing hex&valent cbre>mium content, solubility and presence ef substances 
otli($8> ^ian cbromiunno Because of tbe diffieultf in pr@)ducing carcinonn& 
of the lui&g in experisTtental animals even with kno'st'ni. cardnoget^So the 
giubcut&neous rout<g ai injection <^as selected using a single des® of 10 mg„ 
suspended iu Oc Z ml. of tricaprfllAo To investigate the p^ssibili^ that 
the materials may have an acceleratiisg effect on tbe carcis^genic process 
rather than act ae primary carcinogens they <were administered in the 
vehicle aloo^ and in the vehicle with a marginal dose of a known carcistogen 
Oo 05 mg. of beasofajpyreaeo 

The original <work -was ese^ended by injecting subculaa&eously ii£ mice 
the £@llo^ng chromium compounds using tricaprylisi as a vehicles^ 
calcium chroznate, sintered calciwm chromate and sintered chromium 
triosddeo 

Folio-wing the observa^on of tumors in %-ats in which had been implanted 
calcium chromate in sheep fat, similar iistramuscular implEints were made 
in the thigh o£ miice using chromite roast residue, calciuxn chromate and 
sintered calcium chromaSCo In addition to determimng whether the chromi^^ 
uin ceimpounds are carcis^ogenic to mice as well as t@ rats„ it Is hoped 
that the carcinogenic effect &£ ^e residue eaia be connpared with that of 
the pure chrrnnimn cozn pounds. 

Major Findingss ° » It has been f&und that the fraction of the residue 

composed of particles less thaes 2 microns in diameter 
has ^exavalead chromium contes^t 3 to 4 times thai of the fraction made up 
of particles greater than 10 rinierons in diametero 

The number of tumors which occurred at the site &£ admioielra^on 
among the animals receiving the residue dust without benzpyrene Is 

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Calcisdar Year 19S9 



Significat^ceg 



lo Fx^aetiouaSieo. hj parade size ais,d asal^sis of the residue from 

leachicag coasted chromite ore reveal tlsat the smaller particles 
ceffitaiis a higher pereesitage ^ heacavalesg chromionn that& the larger par° 
ticleso This iecreasea the acute toxicity of the smaller partiele cise 
fraetio&s '^hich reach the luag by iishalatioi&<, 

2o The subeut&aeoae i^ecti@m in mice of 3o4°bei»zpyres)e wi^h the residue 

resulted i^ a lower iacidetace of tumors than whei^ the heE^zpfrese 
was iejeeted alone or wi^.h ver^r imeoluble chromic phoeiphate., The iiicide»ee 
of tumors decreased and the latent period increased with increasiisg am@unte 
of heixavaieiJi;! chromiumo No promotiteg effect was observedo 

3„ Whea admiEdetered bf implas&tatioa im sheep fat the residue produced 

sarcomas in. 4 &£ 10 rats. A inaction of the residue fr@m which most 
of the water soluble compouade had beesa rennoved produced similar tumors 
in 3 of §2 mice when ii%jected 8ubcul^ae4»uelfo No tumors occurred m 
the coatr©! grmips @f the same size. 

4. Evideiice that chromium ia oue or more forms ia responsible f@? 

the increased is^cideace of caiscer ©f the luag im chromate werkere 
furmshed pre^riouel^ hj &e pr@ductioi% of ca&ceirs im rats bf several 
chromium compounds is str@Q.glhesed further bi' the occurrence of caocers 
at ^e site @f implatstatien ir^ mice receivis^ two of these compounds, 
calcium chromate and sintsred calcium chromate. No tumors were 
produced bf eistered chromium trioxid« or sintered calcium chromate 
whes& injected gubcutaneeualf in mice using tricaprylin as a vehicle. 

So By administering calcium chr®male ie gelatin capsules Instead of 
sheep fat„ it h^s been ehct^Ci that the latter is not essential t© ghe 
production of caceers in rats bf chromium compounds, 

6. The results ®f this experiment gire addiHooal evidence that the 

intermediate preducle in the production of chromium chemicals 
and the discarded residue may be hazard@usa 

Proposed Courses ■= ' The few animals ti^t are alive will be observed 
until thef die, but it is expected that the results 
given above will not be changed materiallyo 



PART B Ineludeds YES X NO 



365- 



= 18 » Serial NOo NCI-205fb3| 

Cal.«iftdar Year 1959 



PART Bg 

Pufelicatieass -■'"Accepted £oy pubiiiealdoa fef the A, M<> A„ 

ArcMv@o of Ie<3ue$rial Healths 

Wm„ W, Pafae aiad W. C Hweper, Pr«M3»sxsti©B d Cancers in Mice 
asKd Rats hf Chrosniuxn Compounds, 

Wmo W. Payee aad W, C. Hueper, The Role ©f Roasted Cferoirsit® 

Ore Im tke Pif©duction of Casicer, 



366 



l«*a - Sejrial No, NCJ^ZOSfM) 

Calttotdar Year 1959 



b=4 "Biochemical Studies in Chs-onnium Ca^cerigeaeaia 

Project tejfminated due t@ the depasture ts£ the priacipal 

i&vesligator. 



PART Bs 

PublicatloRe?.»Grogaffi» C„ Ho , Expepimeialal Stu<fie8 in Melal 

Cafficerigeaeeig. XI, The Penetratiesi of Ckromiuxxi 
iato She Cell Nucleus, Cancey» liai95-iE03„ 
{Nov. -Deco ) 1958, 



QC7 



SeriaJNo. NCI- 20S ffe^J 
CaaewdAr Year 19S9 

b»5o Pffoloi^ed Inhalatiee^ of Chx^omic Oxide as&d o£ Zme Ciat&m&ts, 
Barium Ciur ornate and Lead Chnomate b^ Rats 

Objactivess = - Th« cferemium compounds listed are e&me of Hie most widely 

used chsomium pigments to which &a appreciable ©umber 
of werkers hava eoatact by the tespireitorf ¥®ut«» Hati&dlerM ©f ziac chromate 
barium chromal® Aod lead chromate are sue peeled of haviisg an excessive 
liability t& c&atcer of the lu&go Although three of the f©ur compounds listed 
are beitsig it&vestigated f@g> carcinogesdc proper^ee through ^eir pa^eisteral 
admiaist?atioffi b^ va^iovus routes i^ expe?ime«&tal ammals, the tnain homan 
eoatact witfe them ie bf ishalatioK. It appeared^ therefore t© be important 
to titudf the effects of th«ir iii!halatie«& upon the reKpiratory eastern of rate. 

Me^odie Emplo^eds '^ - Sets of fiftf ?ats each ase beiag exposed to the 

iishalatioai of chromium oxide Cehjfome oseide green) 
and of a mixture of equal partu of eii^c ch£'@male (zisc fellow^ barium 
chromate and lead chromate |chr©me fellowL respectively. The pigmesa!® 
used Gotsslet of & miacture of pas-ticle^ of different sises,. the majority of 
which i» kept below 4 microns ie diameter. 

Major Fiodieif s; ° » Although the rats ha^e been exposed for 20 months 

to these pigments »ot atsXf by i»halaU@i&, but also 
by iage«tios a^ skin contact, maey of them are still llvl^go The auccesefuJ 
intrapulmosary lists'© duelioKa of ^e duet used ie indicated by the demonetra^ 
tioe of massive subpleurally loca^d gKeen deposits in rate i^balieg 
chrosnium osideo 

Sigcificamees « The demonetratioQ oi lung cancers in the ammala exposed 

'^ould proid.de importasl evide&ce not only frozn a scientific 
viewpoint but especially als® from a practical viewpoint, since the 
existence of lung cancer hasards from tMs source ^ould be of significance 
to several large industries as well a.» go ^e general public using paints 
a«&d ifsks e@ntainisig these chromium compounds, X» fact, compensation 
claims for such occurrences have become as^ actual issue in this country. 

Proposed Course; ■=■ ° The experiment will be conducted for a maximal 
exposure period of two years. The results of ^e 
experiineiet vvill be evalua.ted after the histological examination of tissues 
is completedo It is hoped that the results obtai»ed are sufficiently 
coi&clusive at that time, that the entire investtgatioi^ on chromium 
c&ncers can be discontinued^ 



PART B Included-- - - - YES NO 



368 



Serial Noo NCI- 206 (al,a2. ,b„c. 

d6, d7. d8„ d9) 
Calenday Yeay 1959 
i. OADR 

2. Eavirornneetal Caace? 
3o Bethesda. Md. 



PHS-NIH 

Iadi\(id\ial Pxtjject Report 
Cslesida? Yaar 1959 



Payt A 



PK^ject Titles ° CagciaogeBic Sfa adies oa Food Additives fc Coafcaaniaaata 

ao Carcioogeaic Studies ea Coffee Sootd 

i. Polycyclic Aromatic Hyds-ocas'boas in Coffee Soot«. 

Principal lOEvasHgator '^ •= Mo Kuralsuae 
Other Investigator - =■ - = W. C. Huepey 

2, Bioassaye of Coffee Soots a.ftd Acetone Extracts of Coffae Soote 

TERMINATED 

Priacipal Isves^gator =• W. C, S^eper 

Other laves tig ator - - = Wm. W. Pajae 

bo Chesnicai Studies or Dairf Waxes (DISCONTINUED^ * 

Principal Investigator » - Jo H„ Seipel 
Other Inrestigator - = = <= W. C. Hueper 

^Due lo the departure &£ the priacipal iiavestigatDr. the work has beea 
discGQtiiaued. No publicaHoaSo 

Co The Localising Action of Berberiige Alkaloids in the Pancreas ^^ 

Principal Investigator ^ = Jo Ho Seipel 
^^Due te the departure of the principal investigator, this etudy terminated 

in 1958. No publicatioaso 
^' Carciaogenic Studies on Aromatic Amines and Food Dyee. 
d-^lo Reactions to Aromatic Aznines contained in Paraffin Pellets and 

Implas&led Into Intastinal Pouches and Urinary Bladder of Doge. ^^^ 

Principal Investigator = » D. Cole 
Other Investigator --.-= W, C. Hueper 

This investigation S^s been termiisatedo No public&tione» 

369 



C„dl„d2,d3.d4„d5„< 
d8.d9^ 
C»le»da3P Year 1959 

d^Z. Biochennicai Isvesfelgaliott oa the Metabolism of the Food Dye, 

Yellow AB iBioaeeay of ITello'w AB and four related compouads 

Priscipal Isivestigator » - W, D oCoaway 

Otfees^ Iisw«8ligat©rs = = E. J„ Lethe©, W„ Co Hueper 

d"3 Aromatic Aaom® Impurities ia Yellew AB ai&d Yellow OB Food DyeSo 

Priscipai lavesligater = - W. D„ Coaway 
Other Is^estjgator* -=.<=--= E. So L.elhc& 

d=.4o Metabolic Stsadiee oa the Food Dye Light Greea SFCFD&C Greesa NO,. 

Psdaeipal laves^gater =■ => Wo Do Coe&way 

)rs - -==» R„ W, F@ldsteis& 



d°§o Metal»olic Studies an^e Df9, Citrus Red NOo 2, 



lor =- =■ Wo Do Coaiwaf 
Other laTesligators <= <=> ° Eo J. Letheo 

d^6o Metal»olic Studies oa the Dye =• Pyridium, 

Principal Is^vestigator ^ ° Wo Do Coe&way 
Other laveetigalore -=' ^ ^ Ro W. Feldsteiia 

d=7o Bioaesa^p @f Aso Dyes and Their Melabolitee by Bladder Implasitati«ffi. 

Priacipal Isv^sligaler ^ <° Wo D. Conway 

Other laveetigatora ° ^ ° Ro Wo Feldsteim, Fc Jo Lel^co 

d^8, A Stud|r of the Carcinogenic Proper^es ef the Fmulsifiero 

PolyoKyelhyleaeC8| Steara^Co 

Principal Xsrestigalers ■^ Wo Co Hue per and 

WkIc W. Payne 

d»9o CarciniOge^&ic Studies on Araesieo 

Principal Investigator «> -^^^ Wo Co Hueper 

Other laveeMgalor =■=-=.-=- Wnio Wo PafEo 



370 



., 3 - 



Serial No.. NGI-206 (ail 
Calemdar Year i959 



Yeasfa fCalesdsy Yaa» 19591 

To«Bi»=--- -<■-- So 6 

Pg-elessiesjai----—^ 3,3 
Others =--- = - _ 2.. 3 



al -^ Polycfdie Arom&tie Hf drocarbos&s ia Roasted Coffee 

Objeclivess - = A previous iffivesSsgation bjr the p^eflcttt authors demons Iraled 

that c©ffee 3@ot ps-@duc@d durieig. the roasliag process of 
coffee beaois c®rat£is&ed a n\;mrib«r @f polyc^^eiic a:?©matic hydrocarbons 
iBLcludiog €areiffi^geme @ae«<, TMa observatioGL suggested that rc»asted 
coffee beas&s also might be comgazninated iwiih such hydroearbieB&s, The^'e-^ 
£@re„ commercial brands of r@asted coffee were ii^vestigated for their 
contend ed aromatic hfdrocarbot&So 

Methods Employeds » -=■ Fous' differseil commercial brands o£ roasted coffee 

were asmlfsed hf chromatographic as&d epectrO'^ 
photometoic K5elh©d®c. 

Fii&dingss ° - Bra^d A which ie a popular braad, moderately dark reasted 

hf the so-called direct roastiag process showed the presesiee 
of 8 meg. of pf^eae &m6 7 mcgo «if flueranlhette ia. I „ 000 gr. sf c@ff ee 

whem sKis'acted ^ith dichiorome^hae^e, and S asd i mcgo of the respective 
comp^uside V7h@i& extracted wi^ hot water., 

- Brand B which is a popular bras&d of iastaat coffee , contais^ed 
2 meg. of pyreiKS ami 3 zncg. of fluoraMheae per loOOO gr. of the snaterialc 

BraQd C which was considerably darker than Brand A aod B in 
appsaraace comlaiaed oo detectable asxsouat of those hydrocarbons -which 

were delected in Brands Ao B, and coffee seots. 

Brai&d D which is a cozsim ercial brand, extremely dark roasted. 
showed the preeejace of pyrene, fluoraothene, benz|a)aBthracene. chryses^e. 
perylene. b@nzoC@|pyre&e, beaso^ai^pyreeseo beiszoCk)fluoranthe&6o and 
benso Ig.hc %j perylene^ Their approsdmate amount in 1, 000 gr., of the 
coffee was l?„ IS, 0,5. 2„ Oo 8„ l^ 4„ 0. 8„ and 4 mcg« respectivelyo 

Sigmflcances - - The level of the aromatic hydrocarbons detected in the 

coffees was relatively low and carcinogenic hydrocarbons 
were not detected hk popular brands A, B» and C, but in brand D which 
has a limited regional consumption and distribution in the southern parts 
of the United StateSo Therefore, it may be concluded that as far as aromatic 
carcinogens are c&ncerEJted, coffee does laot seem to play a significant role 
in human carcinogeaesiSo However, it should be mentioned that coffee 
beans if excessively roasted, can be t^ne of the many sources from which 
polyeyclic arojmatic hydrocarteoiaa , including carcinogenic oaea„ reach 
the digestive organs o 

371 



"' ^ " Serial NOc ,NCI=206|al) 

Calendar Year 1959 

Proposed cousroes » » N© additional expeirimeatal ieveatigationa are 
contemplated at the present timeo 



PART B iQcludeds --YES X NO 



372 



Calead&x Yeai 



PART 



Public&lioass ■■=■ Kurateune, M.. and Huepe,if„ W, Co „ Polycyclic 

Aromatic Hyd^ocarboois in Roasted Coffae 
Accepted for publication ia 3^ No C, ) 



373 



Serial No» NCi-206Ca2 
Caleadsr Year 19S9 



PART B? 

Publicatione; 

Hueper» Wo C and Payne,, WRnio W. 

"Carci&ogetdc Studies on Sool of Coffee Roasting Plantis" 

Accepted for publica'^oa in Archives ef Patholog 



37il 



NGI-206Cb), (cK i^-»S) fteyrainated. No Pay* B 



d=-2 Biochemical Invesiigatien on th« Me^boiism of the Food Dye, 

Yellow AB (Bioassa^ of Yenovi^ AB &nd f@ur related coznpouBds) 

Ofejeetaves; => - When ca??ying out a study to determine -wSsethe? ot n«$ 

long term exposure to a substamce is harmful, it is -p/all 
te> coQsideir Q0t omly the purified substance itself, hut alsos ° 1) impurities; 
present in ^e cemmes-cial materials 2) derived cempouods which may fas 
produced when the substao^ce comes isiito contact ^ith other chemical age?^t,< 
before iagestioa is»to an orgasiiexn ai&d 3) derived compounds, or metabolite 
produced after the original substaace is ioges^edo Cot%sequ@Qf-lf , ia o^t 
inveatigatien of the b&sards which enight be assf^ciated with the use of ^e 
food dye Yellow ABo four simple derivati'^'es as -vsrell as a purified ©arsaple 

, ©f the dye itself are beis^g tested for earciRogeeic activity hj subcutaneous 
iojectioa icato zx\ic&. The chemical luimea of ^e compouade, aloji^g with 
abbreviated terminology and a Qote on their mode fd formatioa are givet^ 

,i{&the follow Bg table; 

' Abbrevia'^ Mode of 

Coii:^ pound tioa Forjsi atioa 

Yellow AB ^l='pheQylaso~2'-Baphthylaz«lae^ 

2=Pheaf lifsaphth{ 1,2] gs-iasole 
1 ° Amliaoo 2»melh3t^laaphlh[ 1 

Ig Z^Oiamieonaphthalene 

2°Melhylnaphlh{ 1 , 2] imidasole 



YA 




PN 


Osdd&tioa of YA 


AM 


Coupiiag of YA wife 




aca^ldehfde 


DA 


Rediac^ve cleavage 




of YA 


MM 


Coupling ctf DA -sith 




acetic acid 



Methedas » » The cowjpousds, as solutions or suspeasioas ia glfcej'iae 

or purified tricaprylia, have beea admimstsred suhcusaEeetis^ 
above the right Mp to G-5? mice^ The experimeat has ao^- he&n ia pvo^re^i 
60 «eeks» Aoimals were divided iato three ®esiess=' cotitrols, a low deise 
Aeries ^series I) s.q.4 a high dose series (eeries lA|o Equal tsunib-ss-s ei 
male a®d female mice were used ia each series « As the asLpeEimetJt 
pregreased, it became necessary to riiediff the injectiOfj schedule, due to 
the toxicity of some of the compouudSc A sumrriary of the dosage schedule 
i9 tabialated belowo 

Total Total d&se 

Series Aoimals DosaEe Schedule 



IA ^ 



20 



Oo 1 KXig. per week for 46 weeks 

Oo 2 mg. every 2 weeks thereafter 

Oo 1 mg. per week for 26 weeks 

4. mg» at intervals of 4 to 8 "we 



So 8 mg. in 52 

iajsctioas 
42. 6 iu 36 

injeeSione 



Controls 60 



Oc 1 mlo of vehicle following same 
schedule &s series I 



^Administration of Compound DA lit, this series was diacoiKtiausd after 40 
weekso A total of 26o 6 mgm^^-sa^ given ia 32 injectaosSo 



Caleuda? Year 1959 



d=2 



In all cases, tha doae "^i^as adnraioistered in 0. 1 mlo of vehicle. Glycerine 
was used as the rehicie ios the first 13 iojectioas at which time it was 
Eiotsd that this zsiaterial produced hematuriac Purified tricapr^lia vcag 
used f&s it^eetiosa 14 aiad thereaftero It vt&e necessary to purify the 
tricaprflin because the cosnmercial material {Eastmaa Kodak) cocstaiiaed 
an impuslly Cprebafeiy caprylic acid) «'hich reacted with i,2<-diazDiao° 
&aphthalen@o aad YellovE' A3, two of the cosnpouads bei&g testedo 



lags? » - Two turners, medullary is appoaraace, lo 5 cm^ ia diameter 
have appeared ia the iserie® 1 mice receiviog compound AM^ 
They appeared after 41 aad 44 weeks, Ho-weves* oae tiimor, 0„ 75 cm. ia 
diameter, znedulla?;^ ia appearance e was fouad m tiie control series after 
44 weekSo The tomos- ^eld ia the AM series i8„ therefore, of a low order 
ol eigtufieasice a-nd ro&j be ia fact due to the vehicleo 

Ajiother interestiag obaervatiooi, which may have practical sig^ 

oificaijce, has heea made in the course of this eKperimeat^ Tha death rale 
for a^znals receiviiig compound AM appears lo be sigoificaatly lower 
th&n. the death- ss.ts tos ces&rol animals aed for all @lher groups under lesto 
This is illustrated iu the followiag tables 

Total Deaths ia First Sixt) 



of Test Period 



Compouiad 




To^l Deaths 






Series 


I 




Series IA 




i iBltially 40 


aolmals) 


( Initially 20 animals) 


Controls 


27 a 




14 a 


AM 


18 






7 


DA 


23 






13 


MN 


24 






a 


FN 


29 






11 


YA 


34 






11 



^ Figures for the eoz^trols were calculated on tha basis of 41 deaths 
iu. 60 cotatrol asisnals^ 

It has also heeu ooled that large abscesses are present at death in 
20% vf the anis»alSo Eighty percent of these abscesses occur in the liver » 
The percentage of animals having abscesses at death is significaatly 
lo^^er in the group of aninnals receiviag compound AM, This is illustrated 
for the Series I aeiiTnais in the following table. 



'7n 



I 



iacideace of Abscesses at Death ia Ses-ies 1 Asis-nale During 

- Siziy 'Weeks of Tef S Pjsiriqd _ ^ ^ ^ _^ 

Compound Dead Aaim&ls vfi%h. Lriver Necroses aaid Abscesses 

^,-^._ f % c^_ jgte-l dead aga^ 

Control s 24 

AM 6 

DA 36 

MN 21 

PN 24 

YA 21 

The ssime treud is pi'eserat its the se^iea lA aaimsils bu$; it is 
less ffiigaific2is,t in. that s@:ries due $9 the lower faumbes* of asadLinalec 

Significaaees = =•- If abacessas a?e the ps-imsry c&use ©f death ia these 
ai&iznala afflicted with such lesioas at death, the lower 
death rate ift the group of aisimals recaiviag compouead AM ma-Y rebuilt 
froit) the pyeveution ©f abiscesses hf this c@tnpo%in4. The agen.t respor^sifeL 
for these abscesses is BOt kaowa to us at psreserato Com p©uR.d AM itself 
©r related eesnpesuada sn&f he useful as d:^ugg in \f®tevius.zy rrssdicins 
a8,d ixk&y also be effective against certaiei humaa diseaseSo 

Pyopo«ed Coulee; - - About 60% of ^e test ai^mals have diedo Ad" 

miBistratioa of the compounds uade? test to the 
remainisf asdmals wHi fee coaliaued ustll 90% of the asjimals Mva died. 
If possible csimpssund AM should be submitted la a suitable ageacf foF 
biological gcreemug fosr bactericidal aud 'riyucidal activitj'. 

EKtei&sioa of bioaesaye lo ©ther epacies is isteaded if the psfajecb 
©a carciBifigenie metabolites of the tw© dyss should be pursued n-ifcre 
e%teii8ivelf at a laics' date^ 



PART B Ittcluded?—--- YES NO X 



o 



77 



d=3„ AstSTsciSitic Araiae Impui-ities isi Yellow AB sa«3 Yallsw OB 
Food Dyea, 

Objec^ves? «■ ° II ia well kiauown tSiat exposur* of lawmaa b<Biifigs I© ceF«.aia 

dj% mtevxnedi&tee yesolts ic tfe« developmeiat ©f bladdes* 
cmacefa Ojse of Haeea iatsr-mediates , bata-ii&pMhylaiyiiae„ Ima b®eia 
»®coga.ise!d £©£• soma lime a@ a sousfce ©f o«cupali©aal bladder casscer. 
Bl&ddes' tumors have sis® beea ps'cdueed it% doge foHo^is^ ^e 6?&1 
adsMiaie^sratiosa of feela^aapfethylaumiBeo 

Two dyee. Yellow AB Cl-pheaylaao-Z^ZMRphlthyl&nnisss) aad Yellov/ OB 
Ci=0-toiylaao-2=aapfet&^lainiael unde? She Barnes FD&C Yeilew NOo 3 
aad FD&C YsUew NOo 4 yespecSively„ wes-s permitted uatil receally as 
food celos'ifi^ge in tbis country^ A Federal ord&F basacdK&g the use of l^ese 
dfcs ia food becaroe effective in May 1959 fFedejral Regi®tes-_24, 883o CI959): 
As of 1955 fee dyes "ss^eire also permitted ia 10 fosreigsi couatries. Tfeey 
ss'e still pe^milted i& the UoS<,A, under the names Ejfiteraal D&C NOo 9 
a®d Essteg'sial D&C S*IOo 10 respecMvel^^ ^®^ use ia extestaal dr^sgs aad 
cosunetisSo 

Although th@ df eg are certified by the Uc So Food and Dsrug Ad= 

miaietyati©® t© coa^ia a total ©f less than 500 ppm. of ia^ermadiateSo 
i^, ia desisratele 4© kaew the actual asmov-nt ©f bata-napMhfl&mii&Q aad pesaib-.- 
oghes" amiaes preeeat in ordes t© properly evaluate the esposiiffe of a 
populatioa to these eheniicalso Although the asssouiat of beta^aaphthylsirniae 
involved hcs'e is small. 16 cosastitates a £ir&ctias& ®£ the total of similayly 
actiffig arosnatic amiiasfi ts> which maa is exposed 4n Ms m©dera etsvii"a!i= 

Tin 



A method ©f aaalf eis has beem developed i^s detsKK-siisiag the total 
a?omatie asBia© content and the bsta-raaphfeh^larniaa coateal of Yellow AB 
asad Yello-w OB food dfeso 

Tea batches ®f dyes obtaiaed fscoan five iadependea* iKaaufaciurers- 
were analy aed« All dyes "weffe oli^aiBed directly Iscs-n the msaufacfe2:rer 
in sealed carloss aad wese pse'viousl^ cerlified by the U„ Sc Food arxd 
Drug Adnni&i8t7ati@s>. 

Methodss = =• Only a brief sttmmary of the aaalytical methed will foe 

given hereo The detailed proceckiffe will fee published ia 
Ih© chemical literature= 

Five g^am® of the dye in ethe? vya® exSig'acted with several p@fflioas 
of 0„ IN HCl at 0®C t© give 250 mL ©f acid eKtracto It is important to 
conduct th® extractioa at 0®Co siace these dyes are hydrolyzed by acid 
with the liberatioa of bcta^naphlhylamiae at higher temperatursao No 
hf drelyslffl secure ttsder the ceiadlti®aa of «he esttractiea at 0®Co 

378 



1^ Ccoot) 

Total Agcimatic Amiae ^ Conteat^'^ 
Five ml« of this attract ri»as aged to ©bteira a measure ©£ ^e total 
ayomaMc &jmia8 eoBteat of the dye^ This involved diaza«4sa«ion witib 
sodiuKnai trite followed fey c®®pliag wdth N°|i=aaphthyl)-el:SiflsnediajTiiKie 
te give a blue celor, the iatcffisitf of which was meaesa^ed at 565 mii usiog 
a BecksxjaK DU 8p«cteeph©tomete3Po From a calibratisB curve prepared 
usi&g at&adard solutions of beta°aapfe^f!amin«o a figUF® fo? the t©«al 
aramatie amine coiateat ^f &e 6f@ %vas calculated ia uaits of beta<° 
;aa phlhylasti ine <, 

Be^-Naphtisylainnioe Content-- " 

The feznaioing 245 mL of acid es&s'act was ev&pofate<S lo diry;?,»90 at; 
rmotm temperature ia a rol^i&is^ vaeuom e<^s.p<»rato7 as^d the rssiduet -was 
fractioEi&ted bf reveraed^'phase celuaxsa parfeities* chsrsmatogrsphy wsisg 
a sfstecfi cosisifitisig of is©butan®l-is®pr©pas5®l-aq.u««3us buffcs" 
|HCl=-Keio pH 1„ 0)o This sfstem was capable of scpa^atinig sssilioeo 
©"loluidiKne aad other axnisies iram ^e £?ee beta^aaph'^hfiamiaa present 
iia the aeid exferact ©£ the dfe„ ' The coacoPits-atiOE of foeta-iiaphthflassiiro.e 
ia the b«t&<=Kaphthflamiiae fractiea was d©ftei*miRed spectrophotome^x'icanf 
fef measiEFemeat of Ihe optical deaaitf at 275 m{i. F^em thiai thes fff«e 
beta-naphthylamiae coalest of tha dfe was calculated, Th« snethod sbi-suld 
b® useful fas' aaalfsis of sSampleo coffitaiai^ a® little as 20 ppmo of 
beta°.naphlhflannicie« 

Findiages =- •= The aaalffical g-eaults obtained are summarised ia TaMe L 
A cempaaiora of Ihe g-esralts ebtaiaed by each of the analytical mstheds 
shows that the majo? aromatic sroaa« impu^afty pyesest Is fe<sta=-napb.- 
thylamiaeo Ab averag® o£ all aasnples iadScatss th&t the bet&^aaph- 
tfeflasnoiae coatemt rep^esasits 85% of ^he t©fcal aromatic aryiises prese-i-T.t, 
la fact„ tha diffg^eaces between t@tal asesnatic amiae CGatent aad beta- 
oaphthylamine coateat are probably not as great as ths results iadieat.e 
because ^e compleaitf ef the. chiromalograpWc method pre seats se^e^e^al 
opportunities £o? los s of imatssialo 



379 



d-3 icont} 
Dye 


Tabla I 


FD&C 
Lot No. 


Serial. HOc WGI-206Cd3| 

Ca.Ie$5!5as." Y®s\r 1.^59 

As-ematic Amisse CoaSeaS^ 
PPMo 




Totei 
Aromatic 

Amiaes 

116 

196 

193 
203 

278^ 
1090 


Beta'»aaphth^lamla« 

hf Chronnato«- 
gE-aphie Method 

@4 

15? 

US 

i?S 

2?4^ 

90S 


FD&C MagBUS„Mafb€e k 
Reyaard 

Yellow Na 3 Bates 

Yellow AB Natioaal AjsiUae 
Kohaatannm 
Nstiooal Acdlirae 
Dykem 


Ho 6589 
K-2699 

G^izn 

W0838 
HT044 
H3604 


FD&C 
Yellow aa 4 
Yellow OB 


Bates 

Kohnst&miB 

Magnus „ Mafbee & 
Re^aard 


H2700 
W0886 
W0094 

W0E48 


8S^ 
131 
i3S 


lOS 

13§ 



Sta8ida?d 
Deviation 



?o2 



10.8 



. All values are averages of two analyses uoless o^erwiae noted. 

Average of th^ee analfses. 
^ Baeed on pooled vsriast^e of all gaitnpleso 



Sigoificance; => «= There iSs at preeeal, jso a^deac® that carciaogerJc 

damage c&&. ^eeull from & long coatiaued exposure t& 
the email amounts of bet&'^aaphthf ia^niae whieh <sfOuid Jt^svdt fscozn tha use 

of these dfes in foods aiad coentQticeo However, there is ax»pie evidence 
that asi occupati€ia&l e^pc^sure l@ l&?geif »H>®u3ts feauita is^ Madde? {iimoise 
iffi a csnside^afele pescesslage of membsf a ®f the population at rieko I® 
view of this it seems wise to exclude €hsse dfes from the f©od aad coa^ 
meticso especiallf lipsfec&eo Cu^aeous coatact "^ith b®ta=>saphthfla3s:isa® 
ie adequa^ fo? producing &n. affective css'Ci^^geoic s^osu?s because 
this chemical fapidi^ peeatrates the skia. 

Proposed Courses =■ - No insthsr sfeidies oa the aromatic amine coateut 
of fellow AB aad yellow OB are coMemplatedo A ' 
manuscript describing 'ihs &ska.l'fUc&l m^biQd in detail has been sulsmitted 
for publicaliGac If future studies are deemed necessary, thef will deal 
with a furthe? studf oi the metabolic degradati^a of the dyes includissg 
bioassay of anf met,abolite8 isolated, 



3Pn..^ 



;v.^c.^^^.,i,-,v.-o..v.^^eig &^ a Fooci ij^e, XAghl C^c^& SFjFDkC Ggees NOc 2} 



.:'ijje&tiveag -- Five ieiphenylraetSajse dy ee ase preseatly approved for 
us© ia feoda bf' ^e U„ S. Food and Dmg AdnniiaiBtsatieffio 
The structures aisd ©amee of these arc littted feelowo 

y \ ^2. ^S / V 




<^£H5 




^<>3 



FD&C Name 



Coznirsas'cial Name 



Gseeffi No. 2 ligM Gre®a SF yeUewlsh ^Q S-/^ \) 



Blue No. 1 Bifilliaot Blue FCF 






Gfees) No^ I Gui&ea Gr«eB B 




// \ 



G^eea No. 3 Fast Graea FCF 



Violei 




HO-// \ 
Na03S 

^^^ /TV 



t^c 



/ Vzz/ 



.9P1 






d-4 CconI) 



Several iadepeodent workeps ^Nelsea e^ al„ , 19S3, 1957? 
Gresso 1955? Hariris^ 1947§ Schaie? . 1937) faava demoaaUated thaS 
subcutaaeous admitustTation of light graea SF pro«Sucea ttumoars »t the 
site of iBJec«ieB» By oral admiais«yati©® of light gsreefi SF, Willhekn 

assd Ivy (1953) wore aMa ta preduc© 1 lympheaaycoma. 

The c&rciaogeme activity of t^ipheoiylmeth&ne df-esftuffe ia uot 
limited to light greea SF„ Nelaea aad coworkers (1953. 1957) have 
produced iajectioa eiie fibrosarccsmae in 16% of 18 rats by aubcutaaeous 
admioi8trati©a of tlie FD&C dyes, green No« 2„ green No. 3, blue No. S,. 
affid violet nOp 1. They oMained oaly one injaclieQ aite fibrosarcoma 
using FD&C greea No, !<. The activity is maoifesft osly whem the dye a 
are administered by the 3ubcuta!&e@us routco 

Large d@ses ^20 mg/week So c, for 2 years) of the dye are saecessagy 
to produce tomorso This may indicate that aa impurity ia the dye is 
the active agents oir the ageiat could be a m@laboli$e formed in small 
amounts Eveia the FD&C dyes, which are relatively pure, conftaia 
several subsidiary dyes as well aa other impuritiesc 

Hess and Fitshugh fl955) were able to recover over 90% of the FDkC 

dyes, blue ao. 1„ greea ffio» 1„ graeia esO« 3 sad violet no. 1, ia the feces 
of rats after oral admimstratiea of the dyes. However, ii& the case td 
FD&C greea Noo 2 flight gree» SF|they -were able to recover oaly 6S%„ 
They could acc&uDt for to 3% of the dyes in the bil© of dogs. None vjs.b 
found in the uris^Co 

Therefor e„ it seemed feasible t© lavestigate the mstaboliam of gre.2a 
No, 2 ^ith the hope of isolating m etabolites of the dye is'hich wcmld 
account for the 32% aol found by Hesa aad Fitshugh. 

Methods Employed? =■ - Part of the ^work of Hess and FitahughCJ. Phs^nn. 

and Exp» Therapeutics. J^,, 3809S5J) on the 
recovery ©f tripheaylmethaae dyes from the excra^ ©f dosed aaiisjals , 
was repeated by us in oa-der to confirm their fiadirags. 

Dye was also adminietered subc utaaeow sly to two rata in os-d^as' m 
determine the relative amounts excreted la uriae and feces. 

Cysteine was administered orally lo o&e animal dosed orally with 
the dyco ia order t© determine the effect oa the escretioa pattera. 

Some of the esperimeals were carried out simultaneoualy usisig 
FD&C violet No. 1 ia order to check our technique. Hess aad Fitahugh 
ware able t® recover 98% of this dye ia the feces of dosed assinjals. 

Findings? = =• Attempts to recover the dyes frem. teat miisturea, prepared 

n on 



"^ ^^ " Serial NOo NCI=E06(d"4) 

CaloBidsur Year 19S9 

hf mixing kaowa amouats of th& dye wilh fresh raft feces, resulted ia 
oaly 50-65% recoves-y usiag Ihe method of Heaa and Fitzhugho Thef were 
able to acfeievs 9®% recovery usisg the pracedure described briefly 
bolows 

Feces are bieaded with water ia a Wari»flr bleadorc 
An aliquot of iMs solution ia diluted '^th water 
acidified with coa, HC 1 „ the solution aatueated with 
sodivim sisifate a&d e^racted with t'^butaaolo Au 
aliquot of the t°butaaol iis t7«»ated wi^ chleraail to 
o^idi^e any of the leuco form of the dye to the celored 
forme A dilutioa of this solution in glacial acetic acid 
is pr@pa7ed and th@ dye coraleM of ^e acid solution 
detarmiiaed spectrophotomtstricallyo 

Uaijag a Eimple modificatiea ©f this method, siamely aubatitutisg 
S mlo of glacial acetic acid iov 2 ml,. ©£ eoaCo HCl in the acidification 
Gt^p, we -^sre able to recover the dyes quatititatively from the test 
£rjif^tu£'®s ^i^ feceso This modified procedura was used to obtain all 

results described hsre» 

The dye ^acovered from rats after adminiateriag 100 mg, sod 
20 mg, doses by stomach lube, in 4 a®d 2 mL ef water respectively, 
ia shown i» Table lo 

Table I 
RecOTsry of TriphsKyimethaae Dyes ia the Feces @f Rats After 
__ _Qrai_ Admittiatratiott 

Rat Doss Period ©f Dye Recovered 

NOo mgo feces ' % 



CollsctioB Green No. 2 Violet Noc I 

Hrso 



100 


6S 


89 


-= = 


1 


22 


102 


„=.« 


i 


71 


===- 


80 


2 


46 


106 


==.= = 


3 20 


4§ 


96 


96 


4 


45 


=,== 


100 


5 


81 


95 


^^-^^ 


Avasage 




98 


92 



No dye was found in the urine. Although Hess and Fitshugh 
found that tha dyes ^sre completely elimiaated fr®m Osborne^MeEdel 
r&tE in 36 hrso ou? observations (Table 11) indicate that a longes* time 

is sometimes required by NIH Black rata. 



383 



Table U 
Rate <af Eliaiia&tioa o£ Violefe Mg» i by ata Adult NIH Blaek Rat 

Rat Noc Pe^isid of Feces Collectioa Dye Recover* 

Hjps. 

29 SO 

53 §0 

70^S 80. 

The elimioa^on tinie is v&s'iaMeo ho^eve?, aad may bs &s shoFt 
as 22 hou^a. 

It is kao'wa tkat tsripheaylmetSiaaa dyes cas bi^d chemically vAth 
protem and it is tfeought- that this blading takes place through mercapft© 
groups im p^oteim. 

Cysteine hydsrecMoride |40 mgo | was ad^nizds'jsred &hing wi^ §0 tng. 

&i gx&en NOo 2 ia aa atteznpl to detei^zniae whether ^e tu's materials reac'- 
OQ passage through ^he digestive tract. A secesd dose c^ <^stei8;,@ h.f^:sc&'- 
cMoride «>aa givea S-1/2 hours late?., Hiaeftf pe^cesBt of the dye was 
recovered in the feees afte? 47 hours <> All of the dj® was ellTnii^^eid in. 
the leucOo colorless iosnio Animals »ot s'@ceivisig cyeteise, dosed eilhe.? 
by the @ral or subcuta^e&us reute, eliiriimite 80 to 90% of the recovei^ed 
dye ia the leuco Sostxia 

This itidic&tes that no reaction product is forxned which ie stable to 
the osddatioQ step used it>. the recovery proeedu^eo It is still possible 
that aa amiao acid moiety is present ia the colorless form of tfea dye 
wMch is present ia the feces. This moiety may ^ea be lost ia the 
recovery procedure.. 

An attempt was also made to produce urinary metabolites o:5' 
greets NOc 2 by adsninisterii&g the dye subcutaaeouslyo Recovery of the 
dye i& the uriae asd feces of the aalmals is sho'«7£s in. table Uh 

Table !U 



Recovery of Greea No . Z Fpllowiag Subcutane<^B Adiiiiaigtr&tioa 
Oote Period ©f Feees Dye Recovered 

Collec^ioa % 

Hrso Feces Uriae 

ii znlo 2% ia water) 

20 mgo 4i 91 Q 

{Z ml„ 1% itt water) 

The dye is largely recovered in the feces The 1% found in the 
uris^e in one case ie probably due to contamii^ti&a of the uti&e <7<;ilh 
fecal materiaL . q , 



- 17 = 

Serial No. NCI-206id>4) 

Calendar Year 19S9 

d^ I coat} 

Visual ohaesv&UQn aad pape? chs-^matography of the chlox-anil 
treated t^butanol e^T&cta (sf the feces of aoimals receiviag the green 
]Moo Z mdicates the prese&ce ef a vielet^ colored zne^bolite. The 
vialet mates-ial w not ae isnpurity in the greesi No„ 2 dye nor is it 
aa artifact i&x'srte4 by interaction of chlorasil with the greein No. 2 
or by inter&elios^ of cMoraoil with fecal coQstitueittSo II is possible 
ho-^eva? that it is a metabolite forzned from oae of the impurities in 
greesa No. Z. 

Paper chg-omatograptoy shows the maia impurity in gre^a Noo 2 
to be Greea Noo 1< Some blue No. I and at least 3 uoddealified mate» 
rials are also ps-eseni is& very small aitiountSo The violet metabolite 
is preaeat iu very email aznouog aad its chemical IdeatificatiQiii does 

sgot seem feasible at the present ti^nso 

Signilisaaees => - The procedure for recovery of tripheaylmethaae 

dyes from €Kcreta of desed a&imals has beea impr&ved, 
With this improved procedure it has beea showa that FD&C greea No. Z 
is n&t metabolized to any greater esttent than other FO&C dyes of 
the triphe&ylxn ethane type. TMs dees not prove thqt green No. 2 is 
insiocuous but oaly eho-^a that the metabolism ef tMs dy@ is not uoiquec 

Siace light greea is excreted with the feces after subcutaneous 
injection^ it ^ould be interesting to know whether it is secreted into 
the i&t@@li»al lumen with the bile or through the intestinal mucosae 
If the latter route is active, light greea Tnay be assumed to e^ter this 
tissue even after oral admiinistrati6n a&d may subsequently be re> 
asEcreted by the same tissue into the lumen. 

Proposed Course; - - Over 90% of the dose of green No. 2 was 

recovered in the feces of rats after oral 

adminietratioffi of ^e dye. Therefore, the search for metabolites of 
the dye would be cos&siderably more difficislt than our anticipation 
bassd on a b$% recovery of the dye by Hess and Fitshugh. In addition. 
th« productioe^ of injection site tuxnors by Nelson and co-workers using 
other triphenyhnethane dyes indicates that green No. 2 is not a unique 
carcinogen asnong ths triphenylmethaae dyestuffs. 

With the techniques prese^Iy available to us. it does not aaem 
feasible to pursue a study of the metabolism of green No. 2. Con« 
sequently, work on this problesn has been discontinued. If at a future 
date more suitable techniques become available or if a more feasible 
appro&eh to ^e study is conceived, work will be resumed oa the problem 

PART B includeds YES NO X 

385 






Sejfiai No« NCI-206(d5> 
CaleiKdar Year 1959 

iftC Sfasdiea oi& th e Dya Citrus Red No, Z 



OfeJecHves; - - CiSrtsiB yed NOo- 2 is &n aso dye having the ati-ucture 
Bhov^n below. It has beea proposed as a subetitvite 
ior FDhC sed No» 32„ also shews below, for use in d^eictg Flo£-ida 
o?&»ge3<. T^ latte? dye has bee» baas^ed because ef its high tosdcitf. 

T-^'o eft^FiiCbUsrallf similar dyes, ©saRge SS aad I°beasea;eazo<°2° 
aapMlislo hsve Iseeji sisowa by Jull and Co-workers te induce tumoya 
wliea implaated itato the bladders o£ miceo All four dyes ob reductive 
cieavag© Vfill produce i»a£nisi0»2<'&aphlhol ^hich has also beee. showe 
to bs cas-cas^geKie I© bladder 8pitbeliunn< Aa isoineric ami!so«iaph6hol, 
aarnelf„ 2°'a£S?is3LO« I '» naphthoic has long beea recognised aa a carcino'^ 
geaic £ne%a.b>oHt3 of beta<° aaphthylaxniae^ 







CitT^iQ Eed. FDkC Red. Orange SS i»Be sef&ea£&> 

No. 2 No. 32 Formerly FD&C 2'a^phthol 

Orange No. 2 

It is ^e object of ^s iavesligatioa to study the znetabolism of 
citrus red No. 2o Metabolites formed, particularly orthO'^ainino- 
Baphtb&ls, v^ll be identified asd tested for carcioogeaic activity^ 

Methodss <- - Cittus red No. 2 {50 $o 100 mg^ ) has beeia administered 
orally to rats. Uriee and feces were collected separately. 

Uachas&ged citrus red was recovered frozn the feces by esstraclioa 
wii h isooctane and deternnined epectrophctoxnetricaliy. 

Urime iRras fractionated by e^Iveat extraction aeid paper chroma^ 
tography. Fractiooa were esamisied by speclrophotoznetric and 
chemical niethsds. 



_•%'=■= Results at this ti^ne are prelimiiaary and coiaclusioQ,» 

ri:&y !i£.'/s to be j^nodified ojs the bt-iis ©f fuluffe 22ip5i.''irrt.s-e; 
Afte? oral adnnimstralidn, 86% of the dose was recovered unchaoged 

ivf '^h? feces withic 48 hours, oOb 



Issidair Year 1959 



d-5 Cconfc> 



The uri^e c»f dosed amrnals is pisk acd givas a positive test 
for a?omatie amiasa. The pink c®lo? is probably raol citrus ?ed itself 
eiccs $he df « is quite itsaoluble ia 'wates*. The cslo? may be due to a 
glucuFocdde ce&jugate oi the dye.. Paper chs'Qzratog?apby of the uri^e 
aeud of extracts @£ the uf iije ie^dicatea the p7eaeffice of seveiral sr.aterials 
iM»t present ia SMrtnal urine. Howeve?, these products have aot yet 
beea identifiedo 

Sigoificances » - Dyea which are ve?y similar structurally t© citrus jred 

NOo Z have b®ea repoTted t@ be carcisiogeidCo A sir^iple 
reductioa product of these dyea and of citrus red itself has also been 
shoves to have carcioogemc properties. The ca^citsogeadc basa^d whieh 
might arise through addition of citrus red NOo 2 to the feuznan diet has aot 
bsen ^oroughly itzvestigattd. The results of this is^veatigalioa "^ill help 
to establish -whether such a hazard esdsta. 

Proposed Course; » •^ This investigation will be costiuued. Uriaary 

metabolites will be ideo^ied a^^d assayed for 
carcinogenic activity. Citrus red itself ^ill be tested f®r cs.rcitiogeD,ic 
activity usi&g the bladder iinplaxstatioa tecbtiique described else'i^heire 
i& ^xis report. 



PART B includeds = - YES NO X 



387 



Serial No. NCI°206Cd^ 
Calendar Year 1959 

d'-$ Metabolic Stadies or Tha D ye Pygjdimn.. 

Objoctiveos -^ = Py?idium is aa aao dye having the following structure; 

The dye h&@ am^lgeeic and aatiaeptxc p?op@?ties and i^s b@ea used fes' 
manf fea^So ei^er as the hydrochloride or in coinbiaatiou with sulfoa^ 

axTiides aad- amtibiotica <> for the treaUsieist of urinary infecliooSo 

Boylasad asid Co-w®rkers (Brit, J. Cancer (1957)212) have shown 
Ihat pyridium will iadwca bladder tamers when cholesterol pellets coa= 
taimag the material are iistroduced intQ tha bladders of xniceo This finding 
casts some doubt o@. the wisdom of uieing thie material for the treatment 
©f chronic uHsiary iafectionSo 

A search of the literature did zmt reveal any nnetabolie studies on 
pyridiuzHo 

It i® the object of this investigation to determis&e the nature and 
quantity of the sirlnary snetabolites produced following oral administration 
of the dye to animals and m&no These metabolites will then be assayed 

far caseisiegQoic activitfo 

Methodss ° °^ritlium hydrochloride in aqueous solution was administered 

to rats via stemach tobe.. Urine and feces were eellected 
separately for two days fdlowing the d&se» Urine was made alkaline ai»3 

es^racted «dth t° butasusl. An aliquot of the buianol esstract i^as diluted vrith 
Oo 1 H HCl and the pyridium centent of this solution determined spectro* 
phot@metricaUyo The butanol estract \n&a als& examined by paper 

chroznatography. 

Findings 3 « => These results are preliminary and may be revised coBksiderab" y 
on the Msis of further esperimentSo 

ApprosdTfTiately 15% of the administered pyridium was eliminated 
In the urine wi^in 21 hourso No farther elimination in the urine was 
detected over am additional 20 hour periods Using the same extraction 
precedurss, 90% rec©very of free pyridium could be obtained from mixtures 
of pyridium and urineo 

Paper chromatography ©f the butanol extract of the urine ©f dosed 
acdmals indicates that none of the excreted pyridium exists in the free 
state« The chromalograms show several colored spots 'which are not 
present in normal urias. None of these spots co^'respond ^ pyridium itself. 

38R 



Galea«3s,sf Year l^'s>9 

Significaaces » =■ if pys-idiuxTi ®r aaf mstaboHle asrisirag feosn it ia 

careioogemc » the jus^ifieatioa of ita fu?tber use in ihe 
treatment @f uTinarf infectioas should be questlonedo The results cf ^his 
iavestigatioa will assist isg dstermisiiiag v/he^e? &s aot & h&z&T<S e:d@ts 
when this eompound is used medici&s.lly<. 

Psopesed Courses ° = The precedure used t© ^ecave? pyridiuro frersi u^ine 

'$9111 be improved so as to obtain a msse accurate 
eetimale of the amouat e£ dye elimiasted ia the uffiaeo Ths amoviBfi elix'nia£-'« 
in the feces "will s.ls© ba determiaedo These figures will ©aabls &.n estiiBaiio . 
lo ba made of th« coQe«s!)t?RUoi% of c^lo^less me^bolites to bs expected 
ia the usrine BKsd faces. Metabolite® ©f the dye will be isolated and identiiiec 
aad assayed for c&rciaogeaic activity.. 



PART B Xncludeds = YES Nq_ 

189 



Serial No. NCI-206id-7| 
C&leada? Tear 1959 

d-?o Bioaeaaf oi Azo d yea and Their MeS&bolitea by Bl adder 



OhjecUvBBt " -^ A zjnethc^d for testiag materials for carciEiogeolc activityo 
by surgical impla&tatioEs of pellets c£»»taieijig the material 
in.to ths hl&^MeTs <si mice^ was developed b^ Jull {1951} aod baa beea used 
udth rotable success siace Uieu by lull, Bowser, Boyla^d and otberSo 

These "s^GskesB have used pellets prepared from either paraffin 
waK o? choleste^olo Both of these substances produce some tumors in 
cpaSJTol E.aims,ls, thereby greatly reducing the sensitivity of the znethedo 
It is mot carlai% ^hatltes- these materials are carcinogeoie ia themselves 
&x vwhethe; they c@^talii a carciaogeoie impurity. Both substances are 
difficialt t© purifyo 

It is the object of this imrestigatiott to prepare aad test several 
pu:s'ified syatlietic v/a^es aod fats t@ determine '«srhether or aiot they are 
@uils.Me ioT use iu the bladder implaatatioa teclsiique. After choosiog 
a @uitabl<i vehicle, the teclmiqae ^U be used t® assay the carciaogeoic 
activity of several dyes st.u.d their degradation products.. 

Methods? =■ = Ths f©llowiag v/ajsea ^TaMe I^ were prepared by reactiotK 
of the necessary alcohols aad acyl chlorides « followed by 

recrystalliaatioa f?om alcoholo 

Table I 



Syathetic Waaees 
Fornnula 




Dodecyl Palmitat.® CjgHsiCOg-Cix Hgg 

Hesadecyl Pabwitate Cj5H3iC02=-Cj^H33 

Dodecyl Steasrate ^lV^3%<^^z'''^-lZ^Z5 

PeUets were prepared from these vsra^es and from tristeariia„ 
a synthelde fat |rr.po §5®C. ^Ic compared ^&i pellets prepared from 

paraffia -wax ^mp, S6®Co } 

Pellets^of the above wa^es were implanted ia the biaddeifof mice 
aiE^d the asinrials observed for a period of 5 moathso 

Pellets were prepared by connpressioa of the po'iardered coi»p@'Uiads 
usiag a stai^ess steel dieo Pellets were tablet^shaped, 3o 2 mm« il/B 
iaehl ia diameter aad lo4 mm» thick and weighed approsdnnately 10 mg„ 

He^adeeyl palmitate was ei»$seQ as the vehicle for large scale 
experim;aritatiQa<. SiKty animals are being inrtplaated with this veMcle. 
It is hopsid that 40 aoimals from this group ■will survive I year ©i* loager. 

390 



=- 23 - 

Serial No, NC1=206 (d?) 
Caleada? Yea? 1959 

Her^decjl p&hxiit&te poUetis. coataisie^ 15% of feho dyee Haled 
in Tafel© II feava fosea prepared aiad are preoeMly baiiag implaBtedo 
SiKtf asiixials are bemg used ia each group. 

Table II 
Dyea Seliag Tested by Biadd eg ImplaafeatJen 

^__^ ^ Chemical Hame^^ , Coiamon Name 

2-C2-znethfl'beiBa@aeaa©|o2=isaphthol Oraage SS , 

i-IE^^-dimefihylfeeiasQaeasel^S^giaphlhol Foymerly FDfcC Red NOo 32 

l-I^.S^dtoethosEfbeasesQaaol-E^naphthol Citrt£8 Red NOp 2 

O^ssge SS waa fouad t© be cascieogeaic by Boaeer and co-workers 
whea tss<t4;d in. ci^oleste^-C'l pelletSo FD&C Red Noo 32 has been m «4 Ui dye 
Flo:k*ida ©raE^gee^ Citrtsa Red Ho^ 2 i» bsitig coaaidered as a replacement; 

ios FD&C Red No, 32o 

Fiadiagss - = Pellets prepared fr®m d«»decyX palxnitate^ hexadecyl pal- 
ntitate as^d dodecfl @t@a?ate are physically similar to those prepared 
fr^rai p&rsdiiu wsl^z. Wh@s& knplasiled in Ihe bladders of mice the pellets 
Tsm&in uQcharaged f<3? periods of at least § tnoi&thSo Animals have not yet 
been observed for logger periodSo Pellet s prepared from tristearii^ere 
judged to h® too fragile for useo . 

Sigadficaraces - ^ The use of hesadecyl palmitate as a vehicle may iiacreaee 

the sensitivity of this method for evaluating carciao^^ 
gesdcity by reducii^ or elimitiating the incidence of bladder tumors in the 
coMrol asiimals^ 

Tha results obtained by testing varieua dyes and their metabolites 
v^iil be helpful in determining what dyes are innocuous to humanso 

Proposed Courses ° » If after further study tUs technique contiiaues to 

appear useful, numerous other compounds will be 
tested. In particular. ortho-aminonaphlliolSc formed en reductive cleavage 
of several FD&C Aao dyes, mil be tested. 



Part B Included^ - YES NO X 



331 



d-So 



Serial NOc NCI=206 (dS) 

CaaendaF Yesr 1959 
A Study of Ihe CasrclKsgeaic Prope^tias ®f the Eznulaifiei?, 

P«tlyos£j^f lessees J Staas-ato. 



Objective®? - = Th® Food Pr©4ec*i©ii Commiitee ©f &e Naliooal R«s«aych 
Cou£icil c&Gcludad m 19S3 INatiomal Academy- of Scieacea^- 
National Research Couii^cil PublicatioKi E80| that "available da^ fail to 
d®moast?ate tb&t polfo^elh^lese stea^ates as-e safe for u@e itx foods undez' 
ail pat^e^Qs of dietary coitaumptiea arad for all segmetsls of th® p&puiatio>». " 

Subs©q«a®3atlf addMasial data weye se^iewad b^ the Subcommittee 
oa Toidcelogy. which rspos'ted CSlAS°Ji|RS Publi cation 646^ "Eve^y aaimal 
is&t} 'Q^t had a Madder ^umo? aleo had at least one bladder stosaco It 
soe^ns likely th&t the tumos'S wes'e caue@d bf chpeoic irrttatios^ of the 
Maddes" epithalium hj the stoaeso " The Subcoxsiinittee concluded „ "the use 
of polf oi£fsthfl®ae|S| etearate at levels asot gffeate? thasa 0. 05 per ceat 
ivk th@ humaa diet %^uld be safeo " it does consider the chemical to be 



TMs p-i'«ject was uadesf^kea to detesrmiae whether the ma:gerial is 
c&^cimig®uic to ami»aie aod wheihe? Maddeir tumors ^thout bladdex* sfonecf. 
could b® produced by ^e administ^atioK of the compouad, To isivestigate 
the possiMlitf <^at the eomp&uad is a p^omotiug aged the e^perimeatal 
pla^ isciuded the issjec^ot^ of the compou&d with a k&o^a ca?cinogeao 

Methods? °-POE(( &} S is fehaissg asSmiassteyed t© aaimals as follo'W8o° 



Feedissg 



2<, Is?;trsmusculas' Injection 

X latrapisus-ai I^jecli®a 
4. latra-pouch Admisai8t&a° 

§c latravesical admimstfa^ 
tion 



60 rata 
60 s>ats 
60 zniee 
60 mice 
60 mice 
30 ?ala 
30 mice 
30 T&ta 

30 ham si 



25% POE«8»S iK food 



10% 

2S% 
10% 
.5% 
100 mgo 



ditto 
ditto 
ditto 
ditto 

i& thigh 



25 mgo in thigh 

50 mgo (repeated monthly) 

400 mgo in gelalia capsule 



SO rate 20 mg« m bladder 



6o SubcutaaeouB lojectioa ia aape of aeck 
\srith 3,4-beaapyretse 

60 mice Oo 025 mg. best^zpfreae & Z$ mgo POEC8)S 

60 mice OoO§ mgo beaapyipeKie & 25 mg„ POE^SI' 

60 mice NO besspyreue aad 2S mgo POE^8)S 

Significance? - ■= This expesiment is a part of this SectioQ^s study of earciao- 
geaic effects of food additiveSo Because poiyoxyethyleae <f8,:, 
aieaifate m&f bs esesS as aa emulsifisr is foods thaS are consumed daily 



Serial NOo NCI-206Cd8) 
Calcadar Year 1959 

tk^aug&cut & pe^soa's lifelisne ■- breado rolls and frozen dairy products > 
i% i@ impos'taat that the?@ should be no doubt ^egardiEig the aa.iety of 
compound w!b@si i&ges^ed. 

Majosf Fi?adinge; <= ° Aaimals tolerate the treatment well but no results 

as yeto 

Proposed Courses - - la tl^se groups which received repeated doses of 
th® material, injections will be cotrtinuad for 

&t least 6 i^oesthSo Th@ feeding ^11 continue for a. total of two years. 



Part B lacludsds - = YES NO 



?,93 



M 



Serial No. NCI=206 Cd9| 
Galeraidar Year 1959 



d--?. CaK-ciaogesic Stodies OEi Arsesdc. 



Objecti^ess » - Alfeho5sgls yepeated effoEta Is* ps^oduee cancera in experimeat.^- 

a^mals hav® beetsi uasuccessfulo tkere is coasiderable 
epide^niologie @.n4 climcai evides^ce which icdicates that arseMc ie c&t'=- 
cin&^etdc i&x masu Recessitlf it has be@s% tabseirved in Europe that vine^rd 
workers ^ho used &sr3@iailc.al pesticide spxay® s.ud who consumed large 
volumes &f ^ia^e contaniirmted by sprays coalmining areeoic experieiace 
a Mgb isacidsace »f caacer of sevasal aites (skia, luQg„ livesrj. Siace 
airsesic may fe« iijgestsd a©t ©nly -wifeh coffitamiaated foods but also ia water 
poilM'Sd by ifiidasgriai wastes c&ataioiag arsesdc, its presei&ee in mail's 
diss is ©I iacreaeij^g couc&r^^ 

Tb® observatioa oa the viaeyas-d «rorkers who coaeuizted araeQicals 

in aa alcoholic soiutioa^ »uggesi-@d tbat the ii&troductioa o£ the chemical 
id tMs i&wrsi might fii3iallf 7es«3lt la a successful production o£ experimental 
arsesElc caaesi-s. Fo? «Ms reasoa the sxperimeatal coaditiona used 
simulate those esaperieaced hy the xi&xk&ra. The followii^g expeomeatal 

sshente was deviseds 

Meth®ds Empl®f®ds « - Arseffiic ia ^e form of As^^O^ is admiaistcrod 

ia drisikiffig waSear to 200 » ci? Black mice and 
£00 Bethesda Black ^atSo Both the rats and mice are divided i^& four 
gs'sups of SO @acb so fhat one group of rats aad oae of mice receive oae 
of the f®llewisg solutisus; 

lo Tap water coataiaiag A@2,Qy 

Z. 1Z% aqueous aoluti<&a of ethfl alcohol c^ataisung Ae^Ojo 

3o !?.% aqueous soluliOQ of ethyl alcoholo 

46 Tap watero 

Tke dose of A^^Oj is beisig gradually i^icreased but is kept to a level thsLt 

pa'oduce® sa® acute SssKicity,. 

Ammals are weighed regularly aad are observed for the preaeace 
of tumors » 

Major Fiffidiagss - - Hone at preseat, 

Sigoificasces ° •» The us&dsrstaadisitg of the role of arsenic i» carciaogeaeeis 

as well as ite role as a food coataumiaa&t would be enhanced 
greatlf if a method could be isiund to produce cancers iu experiznes^tal 

ammals ^th arssaiCo 

Proposed Courses => - The exposure will eoutiaue for a miaisnal esposusre 
period of trw© years if the aoiinala survive, 

PARI' B liiciuae-i.; =• - Y.a.'b^^^ ^ __. WQ_ X 



2. EavJiperameatei Caacer 
Cal«ndasf Yeay 1959 
MedicQlegal„ Conaultative , Advieoigy and Edttc&^ oaal Activities io. 



The 7&pid growlh in. factual knowledge &ti &<a &cti.ial s.nd poten^&i 
exogenous eaasea of husnaa e&&ice?s has bs^ugM £&out aa augnndnted 
aw&re&ss® @£ enviroom eat&l casaee? i%a.3&?ds t@ all epecialif iateFes$:ad 
pariiea^ such &8 members ef the s:n«dic%l prolessi®^. iBdust?i&l snanag@-> 
mesitc i&bor o?ga^salio&s, public hei^ith agencies, znemb^rs of the legal 
p?o£easieQ, birth' coi&trel orgamiza^ons, agric<altu?al &nd faod psoeeesiag 
agencies „ as "t^ell aa to the general p^aMic;. Eavir<g;mn ea^l c&mz&T hazards 
aad ca&c€Fa h&ve became a recognised public health pro^iezn of distlact 
stature. The nisnf aad incre&aiagly freqae^t inquiries on €his subject 
made bf intragoverameatal aad estragoverB.?r?entaic, public aad private 
parlies to th« Envirosamealal Caacer SecUen ?eq»i5fe a ceestaat collec» 
tioa o£ per^iaeiat data £?©m the literature and thai? c^icir^peteat iQterpreta" 
tion in the li^t of the old accusnulaled and eiswlf acquired kno^ledge^ 



M&Q Years ICalet^dar Year 1 

Total = =. = » „ iO 

Prdfessioaal =><' = = .03 
Other =<.=.<= ,07 



Medicolegal Activilieas 



ConsultaMve cooperaliea with the Bureau of Empleyees' Compeaaatio: 
Uo S» DepartJonesat ef Labor, was coati&ued ^ith the esceptios d a brisf 
period during which I withdrew from ttiese activities because of temporary 
ceasership applied t© two medicolegal papers prepared hf me for pubHca- 



Aesistaace haa bees givea oa several ©ccasloas t© msr^bera of the 
Food and Drug Admiaistradoa oa problems ©f caacer hasards from food 
additives aad coislainiaaats aad drugSo They were supplied with repriats 
dealing with sueh mattsrs,, iacludicg copies of the still unpublished paper, 
"Conaimier Goods aad Caacer Kaz£rds"o siace it gives the most complete 
amd competeol review of ^sie importaat problem, including act escteiasivs 



395 



= 2 " Serial No„ NCI=209 

CaleBdar Year 1959 

Medieelegal advice was given to & mxmb&r &{ attorneys in coanectioa 

with Utig&UQ^B cm t?au3Tisitic aad occupational cancer ciainnso Likewiseto 
free e^p&st teeUrsiouf was offered mpoQ request t& several industrial 
coTs^ces-as on queslioi^ of eceupa^onal cas&cer hazards and eompe^ealioa 
suits asad to several private par'deSo sueh as the Board of fehe State 
Um^®^Bitf of Iowa, in regard to cancer hazards from food additives and 
ce^taxnism^atao Because of fundamenlai differeaceg in social and scieolific 
philoaophy^ I resigned as cor^sultajat £r@m the Wax Committee ©f the 
Amerieaa Petroleum Institute. Coofe^eaeee were held with members 
©f the Welfare aad Compeisisatioa DepartmesBte of the AFL^CIO as well 
a® vdth &e E:s£ec^tive Couacil of this orgaoisation on the scope aad naltire 
of occupatioE^l caaeer hazards i«i industry and otb measures indicated and 
needed for theig' conlroL Members ©f Congress were supplied on request 
with literature pertaiaing to questions ©f cancer hasards from f©od addi» 
fives aad coatsmii^ats and of a prevesative control ©f envirenmental 
cancers. 

SigEsificassces 

The activities cmtlined are eeseratiallf of an educative and iofonna'^ 
tive sialureo It is hoped that thef may aid in alertly an increasit^ number 
of individuals to the serious threat to public health created by some 
products of ssiodern i»dustryo 

ProepecMve Cowrses 

The past activities in this field ^^ill be continued whenever they 
are requested or ikeede6o 



Publicatioaas 



Hueper, W, C. „ Laftverunreisiiguag und Krebs ia Krebaforschung und 

Krababekaropfung ., Volo I1I„ Maachen= Berlin, Urban aad 
Scfewarsenberg. 19§9„ pp. 162-189« 

Hueper„ W, C. , Prevention of Occupational Cancer Hazards. CA=BulL 
of Ca. Progresso 9s88°97, ^May-June) l9§9o 

Hueper, W. C „ Eavironmeatal Cancer = Definition and Signific&Kce of 

Environmental Cancer® in The Physiopathology of Caacer , 2nd 
ed. Fo Homburger„ Edo , Part 121, Chap, 24o New York, 
Hoeber^Ha-rper, 1959, ppo 919«9?0. 



388 



Public&lioBfl (con«)s "' ^ 

Hueper. W, C „ Epidemi«l©gic, Expes-imeatal aeid Histological Sfeidies 

oa Metal Caace^s of the Luago ACTA Uoiosi IaS;e:rn&tiossale coatre 
le caQce?o XIs424=436. i959o 

Hueper, W„ C. , Tg&uma and Caacgr ia Ty&mna . MoHoats^ ed<. „ VoL 1, 
Albaay, No Yo „ Matthew Besder & C©« „ Inc. 1959. pp. 47<»li0o 

Speaking Essgagemetits g 

lo Sixth Scieslific Meetlsg held Berlia,, Geymaaf, March 12, 1959o 
of Deatech«r Zentsralaussshues fur Krebsbek&mpfuag uad Kirabs^ 
foJTschuag e. V. (Germao Ceotral Comnnissioa ios Caiace? Co&tfol aad 
Caocef Research) bf invitatio& of Pffofo Dr» med« R MartiaS fPreSo ) = 
Subjects => "Air Poilutaats aad Cance? ©f the LuQg. " 

j:„ By iavitatiosi of Arbeitsgesmaiaaehafl fU? Ks'sbabekampfuog des l^&udea 
Niede78achseR, Ha^aover, Germaa^, March I§„ i9S9o 
Subjects ~ "Air Poiluta&ta aod Ca&ce? of ^he lAing". 

3o By i&vitatiOQ of the Director „ D?. Oo MilMbock. Netherlands Case es- 
ItMtituteo Asm sgterdarn „ Mafch 16, 19§9. asd Maych l?, 1959« 
Subject; = "Exparimesslal Metal CaiseeFs" sad "Caeseea Produced 

by Water Soluble aad InsoMbie Polyisiers"« 

4o By invitatien of the Deutsche Foyschungsgen^eieschaft (Ges-maa Bye 
ComnnissioEtland OccupatioGsI Caace? CaznRiissioa„ Bad Godes^rg, 
March 18, 1959, 
Subjectss <^ "ladustriebediegte LitftverunreiniguQgea uisd Krebs" aad 



5, XevitatioKi of Masiagemeat &i Bayer'isperke, Leverkssaao Mai-ch i9„ 1959. 

Subjects = "Study of Industrial GaBcer Hasarda ia the UcS„ A."be;iC!S'« 
the Safety CouaciL 

6, Participation in Goaferettce Deutsche Ferechusigagemeiaschaft^^Dye 

C@tnznis3ie>n) on Cauicer Kasards from Food Additives and Dyes. 
Ii&vitatio^ Profo GrosSc Bad Godaeberg. March 20, 1959o 

7o Conference with Prof, Go Kahlau„ Fat hole gische® Insti&it, aad 

Dr, medo WittgemSo Arbeitsschutaarst d, Bu&desbah&^^Sosialarrit, 
Frankfurt &m Maia, osi laiug Ca&cer Haa^rds Ameng Ziac Chromaie 
Spray Painters, 

8, By invitatiof!L of Dr^ Frederick P. Bornsteiao Chs^ri, , El Paso Cou&ty 
Medical Society, Te^as, April i4„ I' 
Subjecti =• "Environrn^itai Caacer". 

337 



Caleii^as" Yea^ i9»; 



9o Bf invitatioa Ds-. James Crabferee, Deao, Public Health Semiaar » 
Graduate School Public Health, Utdvesaitj of PlUafourgh„ April 
29„ 1959, 
Subjects- "-EnviFonrneatal Calces- I^sards as Public Health Problem s". 

iOo "Experimenial Cancers ia Rats Ps-educed by Ch?omiiam CoxRpduftds 

and Their Sigmficaace t© Iiadustry and Public HealCh" preseoted bs£o?e 
tis« Anaual CoaveiitioQ of the Americaa Industrial Hygieae Associatioa, 
Chicagfs, IlUeoie, Apsril 30c 

IL Aaaual address before course iu Adult Health at Yale University 

by Xavitation of Difo Eo M, Coharfto May 4„ New Haveao ConB.. 
Subjects =■ "Esxvirorameutal Carcinogens. " 

12= By invitatioa of Leo Perlia„ Dir. Gosninuisity Service Activities „ 

National American. FederaHou of Ltabor aad Congress d£ Jadustrial 
Orgaaisationa, Hotel Statler„ WasMugtoa, D. C. May 25, 1939., 
Subjects^ "Prev%%ti@is of Occupational Cascers <» A Challenge to 
Public Health Ageucies, Mdustrial Mauagemeat and Organised L^bo?" 
Paper cleared for publication iu PrcCc of General Meeting of Comzni^: 
on Cominunity Services „ AFL=CIOo 



i3o By issvitatioffi of PresideBt, AFIi=CIO {GeOo Meaisy) before Essecwtive 
Councilo AFL.=CIO„ August 20 „ Uoity House, Forest Park, Pa« 
Subject; - "Occupa^onal Cancer &n6 Orgamsed Labor"o 

14, Research Paper,, "Career in Chrosmate Workers" before the Semisssr 
Some of the Causes of Cancers ia Mas and Mous<^ Dr. C G. Zubsod, 
Chm, HCi Clisdcal Staff Meetiago Cliaical Center Auditorlunn„ 
Bethesda. Decesnber 10, 



Honors aad Awards g 

Elected Fellow Americaai College Preventive Medicine? Member „ 
College of Americas Pathologists » 

Presentation of Aanual AAAS'-Anne Fraiakel Rosenthal Memorial Awardo 
Annual AAAS meetingo Chicago„ Deceuber 26, 1959o Will be 
present to accept award. 



Serial No, NCI=-.210(&; 
h.. 
Calsadar Yeas- 1959 
PHS^NIH 
Individual Projeefe Report 
Caleada? Yeay 19S9 



Part A, 



Ps-oject Titles ° Sfaadies on Occu patioBAl affid Eavigoaroaatal Reispiga.tog'g 
Gaaeer Haaayds Caaaed by the la halalioa of'Cagboaaceoiis Aig P&llufe^ata 
fCoal Tag aad Petroleum Asph&lt F ames, Cufefelag Oil Fog. Roasted 
C offee Soot Dust) aad by ^ e ImplaBtsti®a of meael Eagiae So ot,, Cigarane 
Tag aad 3, 4'°Besaapyggtte into an Art ificial Sgoaghial Cyst» 

a. Inhalation gf Carboaaceeus Air PoIlutaBfes 

Pyincipal Inveatigatoar - => W. C. Hiseper 
OSher lovestigator -=. = --=- Wm. W. Pafae 

io Coke ©van tae and petrelsum aaphait 

2, Cutting Oils 

3, Co££ea Roast Soot 

b, Urbaa Air Pollutant Study 

Psiaeipal Investigator • - W, C. Kwepe? 
Other lavealigator ----- Wm. W„ Payae 

Cooperating UBites ° - M?o St era and Mr, Tab©r„ Taft Saoitax-f 

EagiaeeriEg Ceuterj Dr. P. KotiE„ Uaivejrsiiy 
of SoutberaCsliforoiao 

c- Implanta UeR of Diesel Engine Sootp Cigarette Tag aad 3,4-=;BeaapTygeae 
JBto an Arlifiei&i Bgoa cMal Cyst of Dogs 

Principal lavesligaSor ° ■» A. Keteaum 

Since cigarette tar caaisot aay ioeger be obeaiaed 
fffom our former source for contisuiog this experimefit;, 
the fui-ther observatioa of the treated dogs has bees». 
placed mth Dr„ Ketcbum ^hOo I presu2ne„ vnll report 
©B tfee further course of tMs esperimeat. 

Maa Years ^calendar year 1959^ 
Tolal=- = ------=- Z.IZ 

Professional-""-' » 04 
Other-" 2„08 



3S9 



Si. Issla&latioB of C^.s-boascesaus Aisr Pollut&ats 

Cfejeetivesi ■= Tlas al&ymisxg rise of luiag €&acera durisag rac«?Sit dscades 

has directed attention to conm bus Ilea and o^daties psroducts 
«(f vaHeus Ifpgs fsC caip|»®aac«©tts msner |c©&l a<&oe. coal tasr. pitch, 
carboa blacka, pcii^oleuira aephalfe, cutttog ©ils, diesai e.ad gasolise engine 
essfeauata, ta?ffy f«m©« fsrom tobacco products, sooly rziattsrials froirj coffee 
ifsastiag, ©jd4aSi©Q assd polymerizaMoB products cjf vsIatiiSaed gasoliae) 
-^sMch p®Uul« t® aa isc^eaeiag degree Ihe occwpa^ieasl aad general huanas. 
eraviroameato The sfeadf ®.f coal ta.s aod petr©l«um asphalt Creofiag, road) 
fiimei§, cuttiag oil fogs, as&d c©ffee roast aoet are impsrlaaft ©ccupatioaal 
seapir&t&sf csirciaogsas wMcJs have attracted lim<a attealiea by ©■feer 
iaveiBtiga^rSo Co£f®e r©a»t soot also -m&y play a c©a«ribut®ry but noiutor 
r©le ia accouatiag for the astoaioMnglf Mgli luag aad laryns: caacer 
morbidity aad mortality rales in. Ne'w Orloatas v^feere c©ffee reaatiag 
plaals aad air poUuMom caused by their e£flu@ats are eaviroameatallf 
uaique features, 

IK Coke ovea tar agid petre le mra asphalt 

Methods Empi©yed2 ^ - Coke ®vea coal tar aad petrO'leunn roofing asphaU 

are healed b^ ara electric device within exposure 
cimmbers to 27§®C» fco 320OC. , res pecfdveif » Large amewats of fumes 
are Iherebf givea off by these zna^erialSo The temperature wM&ia the 
chambers is kept at 7&® to ?a° F. with the help of C(»ld air delivered isE.t© 
the chaznbes'B frozxi aa a£r->eoaditioaer„ Rats atod guinea pigs are used &s 
expermieats^l agdmals "W^hich are exposed 5 da-fs a week f@r aboiit 6 » 7 
hoars each dajr o 

The carciaogeaic poteacy of the materials used is determined hj thn 
repealed cutaaemis applica^oa of the c@al tar aad pefcsroleuni aaphaU %e 
C57 black mice aad by the iatramascular injec^oa of a Elter ositract 
©btaiaed fr©m materials which have passed the exposure chambers aad 
have foeea retaiaed ia a fiber glass filter, 

Fiadiagss ° » F«?nes £r@m healed c®al tar and petr©leum rsslisg asphalts 
did aot pr@d?ac© casjcers ©f the Miags ia rats and g^sinea pigs 
iahaliag each lames f@r a period @£ up t@ 2 fears, alth.&j.gh the admiQistTv%^ 
tioa of coadeaeates of the coal tar fumes to the skia aad muscle tissue 
of mice desnoastrated the high earciaogeaic potency of this matter« Th's 
roofiag asphalt tested proved to ba Boacarcij3©geaic vnheu applied to the 
skim td mice a ad rabbits. |See Table 1}. 

Sigaificaaces ° ° The obsierva^oas made add to the m^uatiag evider:s.ce 

showia.g that aaimal species usaallf used is iahalatioa 
expcrimeatfi of earciaogeaic air p^Uataats prove t© be refractory^ thereby 
iadieati«gg that the test sriethod employed ia its present form is UE^sisitabie 
for the purpose is^eadedo 

Part B lacladed = — - - YE3___^__ ^9,»_.«« 

ii nn 




g 

IS 

»»• 



m 



^ g; o ^ 
ii ^ 

(D 



•1* 



(8 






us 









'd 



O 

c 

o e 



© 



f6>^ 



2. ^ 



i/i 






*^ 



o 



CO 



r 



401 



^;aI3a^i^ Yea? 1 9S9 
PART B s 

PubiiceUeas; » » Haapesr^ W. C, and Pajrne, Wm» W. , Carcissogemc 

Studies oa Petselaum Asphalt, Cooliag Oil 
and Coal Tas-^ Accepted for puMicatieffi 
in Agchivee of Paftholejgr^ 



ii02 



C&leadar Ysas- i9S9 
a«2 ■^-Cttltiag Qila 

Methods Emplof «ds => = Cutliag ©ij|epra«eatiag a miscluffe ©f parafiin ©iJ 

wi1& las-d/J as injected sa a l®g into ^e exposure 
chamber l>y a Jet' operated with compressed aiif and idet&^eal with jeSa 
employed in industry for the appUcatioa of cufttiiag oil®. Bethesda black 
?ata aRd guiaea pigs are ejqsosed t© this fof f ok- about 12 houys five tirmea 
a -sveekc The pas-^ifia ©il fraction of '^e cuStiag oil is„ mo^eove^^ paioted 
©a the «kia of mice ae well as iratramusculas-ly injected. 

Findings s = -The mice painted op iojeeted vei^h th© paraffia ©11 fracMoja ox 

the cuttiag ©il developed ia five ®f the 1 50 araimaSs culaneous 
neoplasms {l papillG>ma„ 2 caffcinsma aad 6 leakemiao). The autopsy 
aad histologic essamioaMea of the gi^sea pigs a:©d rats revealed the abserace 
©f tumos-e of the luagc. ^Table Z}. ' 

Sigaificances - • While the evidence on haod shows that &e p&raCfJa oil 

liractioiii is mildly carcisacgeaic to ^he skia of mice, the 
gross evidence oe ?a|s a^d guinea pigs isrMch iishaled a fog of a ble^d of 
paraifita oil at^d lard/T'S negativeo %eve?$helesa the rapidly spreading 
practice ©f applfiag csstSiag oils in the fosin of a f®g dese^vee coatiaued 
se?i@us altestion because ef the sa&ura^oa of ^e ais* i» meSalltsvgical 
eotabliffihmeal:® with a lively dispersed ca^ciiiogeaic oil v?Mch may create 
a aigaificaut cascer h&m&s^ to the variot&s T<sispi^&^.Qrf @?g@»e &k well as 
the skis, of saaso "whe^e csaces's fs-cm a^cfe a source have recently bsesx 
placed OQ ^scord ia ^aois^tss couBlrie«s is^.cludiag the Umted Stateso 

PART B Included YES X NO 



a°3 ■'^Coffee Roact S®el 

Methods Employed? - = T^e types ®f coffee yoaal soot wes'e <fetalaed iron: 

coffee s'oastiBg pl&ata is New Oifleaas thyough the 
cooperatioi» of the Sagjltasy Eagiaee^iag Caate?, Giacisassati^ Ohi©= Oae 
came frozn smokestacks of plants ueisig aa iisidisrect methisd of ?g»a6^iag^ 
io e= „ a gas flam® heated the ouSside ©» a rotatiag metal drum coataiisiag 
the coffee beaiBS? ^Mle the other soot ■xa^as recovered from a direct roasting 
procesffl, io eo , a gas jet iEfrodwcod into the inaide of a rotating drtvm 
provides the roaatiag heaS^ The soot ie pulverised to proper particle sine 
ia a micr^pulveriser cooled by a dry iee pack. This fiaely po's^dered 
material is agitated ia a flask by a hammer operated by aa electric mcto>: 
aad the dust cl@ud forn;ied is blo^^n ii^o the exposure chaxr.ber, Bethesda 
black rats and guinea pigs are used as ercperime^tal asiimals. About 130 
to ISO grams of soot are used dailyo 



403 



■tii 



•4 {B 



O 



CO 



C8 






£ s> 






O 



o o 



^ 



ml 



8>J 



G 



ilOif 



Calea^a? Year 19§9 



Fublicationss =■ => Kiaepes's W. C snd Paysae, Win, V/<, , Caffcisaogealc 

StucMes Oft Petroleum AspSialt, CoQiing Oil 
and Coal T&r. Accepted for puMicalio2% 



405 



- 6 - Serial No. NCI-210ia3| 

Caleada? Yeas 1959 

Sigaificafficeg - - Hiffit©l©gic studies sSiowed thstt pE'ccaacerous leeioas 

&s%d beaigQ £ad maHgi&aiaS sie«i>plassns of the ekin^ 
eonaeetive tiasueo stomach &nd biadde? dev@lapii%g i» snic@. irats aed 
guinea pigs respectively, fellowirag thais esspoaure to soot f^om coffee 
?oas$ii!SK pl%«ats or its residue of acetone extract bf the cutaaeous, intra- 
muscular at^d respir&lsirf routes attest to the carciaogenie property &t 
this air pollutaat. 

The eccurreace of cystitis cf stie&o papillomas aud ca^ciaomas in 
the bladders of guinea pigs ieahaiiag aad iogestiag finely p^t^n^dered soo^ 
from coffee roasting pls.nts make this material suspect oi ccrataisiog a 

carciaogera or carciiiog^iias excreted in the urinso 

From a critical analysis of experitnentai aud epidemiologic data, 
it is sjot likely that effluents from coffee roasting plasits in New Orleans 
are a major causal fac^r of the unusually high mortality and morbidity 
rates of cancers of the lung, larynx and bladder observed amosig the 
resident populatiem of this city. It is, oa the other hand, possible that 
this air pollutant assuixses a contributory role in this respect. 

Findings^ ■= => The au^psy observation of the guinea pigs aho^'ed the 

presence of nodular or flat tumo?s of ^e Madder in b?m 
anitnalso One of ^ese tumors was a spis&dle cell sarcosna and a &qmixnf>u3 
cell carcinomao the eecoc£,d ^^as a squamo^as cell papilloma. The r&t$ 
displayed tumors of the liver ia S animals „ tumors of the uterus in Z^ 
and doub^ul tumors dt° the bladder and skia in one rat each {Table 3). 

Proposed Courses -^ « The experiment has been terminated for the 
"•*^ present time. 



PART B Included .----YES--J5=- NO== 



b '- Urban Air Pollutant Study 

Objectives^ ■= « The phenomenal increase ^ luag career amoag urbaa 

populations and the distinct irregularity in the deg?ee of 
such an increase in different imetr«ipolitan areas provide circumstantial 
evidence incriminalii^ environmental factors related to industrial 
activities, traffic and transportation eonditioi&s , fuel consumptioiac dispose'] 
of municipal 'wastes by incineration, and similar factors of the mcd<sz» 
economy in the causation of lung cancers and in their rise in f^equeacy. 
After being approached by Dt. Fisher for assumiBg direction of the studies 
on lung casscer as a part of the Ais Pollution Program as a iisst step ia 



406 



Gaieaday Yeaa- 1959 



Tabl« 3 



Reactions in. Mice and Rats Aitew Cu%&.&90%z@ Appiicati©si„ lutramuaculap 
Injection 01? Inhai&tioQ of Coffee Soo^ 9j&4 Rssidue of Acetone Extract 
Of Coffee Sooto 



Material NOo Sur° 

&n6 ?oule of viv@?s 

of adiBlQ" Am-^ over 

istratioa Species mais i yg. 



Direct soot Mouss 

Acetone 

Residue 



50 



ladirect 
ooot 
Aeeteae 
Residue 
Paiotiis ij 



Mouse §0 



36 



40 



Direct soot Mouse 50 
Acetone Residue 

iRtram us eula r 



35 



Indirect 
aoot 

Acetone residue 
ifilr amus cula r 



Mouse 5G 



Direct 8@ot Rat 

Acetone Residue 

Intramus cuiar 



30 



Indirect 
Ace^ne Residue 
Intramus cular 



30 



Indirect 
Soot 

Intramus cula r 



Rat 



30 



Tumors 



isOina 



I leukemia l;^mphoid 



2 Ifrnpfeomas 

1 fibrosa7cozTia(&igh) 



Othe£' 

Leaioaa 

? pidarmal 
hyperplasia 



Epidermal 
hyperplasia 



20 i squasnous cell 

carcinoma uterus 
2 adenofibromas 
breast 

1 choiangioma 

24 1 anaplastic carcii^oma 

stomach 

1 fibrosarcoma thigh 

2 adenocarcinoma uterus 

3 ade&efibrosnaa breast 



foreign body„ 
feasn cell 
graaalomas at 

^ site 

1 callous gas'i; 
ulce? in 
glandular 
mucosa 



22 1 rouQd cell sarcoma 

ileocecal stodes 
2 reticulum cell 

sarcomas liver 
I choiangioma 



Multicystic 
foam csll I 
giant, cell 

granulomas 
in ^iieh 



407 



6-a (cocsti Serial No, NCI=210fa3| 

Table 3 (cfentj CaicBdar Vear 1959 
Re&ctiotis in Mica aad Rats x^fter Cutaaeaus Applicatioa, lolramusculas: 
Injsctioa or lobalaftion of Coffise Soot ai&d Residue ^ Acetone Estz^act 

^ Coffee Soot, 



Mates-ial 
of adsiiiE%» 



istratiea Specieg 



No. Sur<= 

of vivo?8 

Ani" ore? 

znals 1 ^?o 



Tusr @rs 



Othei- 

LiesioQS 



Coffee 



Rat 



60 51 






Coffee 



Guisiea 
Pig 



25 



25 



Soot 

DiE-ect aad 
Ii&di?ect 
iahalatioR 



3 adeQ©!natoeia„luag 
8 reticulum cell aar° 

cQxna liver 
i squamous cell car> 

cinoma akiaChead) 
1 adeaoca?ciBoma 

ute?u8 
Z chala{^{ionnas 
1 cf stadeaotna ovary 

4 adea«fibroiTia breast 



1 purulent 

chronic cyelitifi 
ulcerative 
3 calcifications 

of elaetic 

tissue luag 
1 glacdular 

hyperplasia 

uteruc 



Z adeoL^matosis lung 
2 careiaosnas bladder 
1 papilloma bladder 



6 broaehlelar 
ial 
la 
2 calciflcatioa 

of elastic 

tissue luag 
1 calciflcatioa 

muscularis 

stomach 
4 cystitis 

cystic a 



408 



Serial NOo NCI--£iOC&3D 
Caleadar Year 1959 



Part B; 

PuMicali©aes = - Huepe^, Wo C, and PafT£®„ Wm, W» , Carcii&eg^Edc 

Studies @f Soot of Coffee Roasting Pla&te<> 
Accepted f of publication in Archives taf 



ir no 



iMs task a plaa -^vas evolved i&v cos-ffalaSiJzg epidemiologic data available 
©a the lung cance? iacideace ©I 8 met?@peliterj. &res.s vdth. the data ob- 
t&iaad by a chemical anaifsis of the particulate phafe© ©£ ais- poUutatits 
£?@zn these areas and "with data to be aecueed cos.cermsj.g fehe earciixogeaic 
poteacy of this phaas aad seveyal fgactieas thereof ^»hea tesfesd ia mic® 
hy their subcutaneous isnjecti^n aad compared with a staad&;d e>i 
eaacereue f espouses te graduated d&ses of 3„4»b3uspyrei%e a<imi&^iste?ed 
to xnice bf the same p^ute. 

Methods Einplof"edg =■ <- Becauis® ©f krao-wa differences ia the fs-equeacj? 

^ates of luag caace? it&the papulaM&ao <^ certain 
onetrepoiitaa areas &nd ol differences in the chennical eompositioa oi the 
particulate phase of their air pollutants^ the Saoitary Ei;i@i&eeriag Center 
t@ok charge of the ccUsctioa aad chesnieal analysis of the air po&ttas&ts 
in the following 9 snetrapelilaa areass ■^ Los Aogeles , San FraaciecOc, 
Nashville, New Orleaa®, Birmi^hai7!„ Ciacii&natio Detr&it, Atlanta, 
Philadelphiao Nashvilla was added to the origits&l 3 cilieai in December, 
19i8« It was agreed t& test hf snoathlf isyecHoia iato C57 black mice, 
standardised amounts of the total particulate phase, of i^s crude beasol 
e^racto ils refisied aroznatic fractions its o3ddized aromatic fraction, 
and ita aliphatic fraetioao The bioassa'^a of the first three fractions w^s& 
to be performed by the Esjviromnental Caiicer Seetl^n, while the last, tr^i^i 
fractions were t© be studied hy Dt<, ¥. Kctin ia LjOB Angeles, accmi^disg 
lo methods agreed upon, Bioassay of the tetal particulate phase l^s 
become pssssible ©alf eiaee Decenibar 1958 -whea sMs iTsa^erial was 
collected ©a millip@re filters instead ol glass fiber filteroo 

Each fractiea is iejected into 72 mice in monthly intervals vd'dr 
standardized amounts of rnaterials collected aod analysed durieg the 
prscedis^ monlho 

Findisgss - =Tuinora asm ©ccurring isi mice receiving the eztracts iz&sn 

the air pollutants of the 8 cities as well as tho®i receiving 
graduated deses of beaspf renec The number of tuznors that have occurred 
ia each gr©up is tabulated bele^sfsCTable® 1 and Z)„ 

SS, gnificances - » The number of caacerous reactions ia the various 

series at the site of injection of air pollutant fractions, 
while not eonsiderableo never thelesa is significant fer several reasons „ 
The observations made indicate that there esdst apparently definite 
differences in the quanl^tstive as well as qualitative comiposition of selveet 
soluble fractionfl of air pollutants for different cities as far as their 
carcis&ogenic potency is cancernedo It la evident that the main carcinogen- 
potency ®f these fractions is present ia the two fractions coat^iaiag 
aromatic compounds.. On tfce other hande the maximal tumor yield i&r 
fractions ols^ined from air pollutants collected in Lkss Angeles and 
New Orleans was f®und for the aliphatic fraction, suggesting that possibly 



Serial No. NCI=Ei 
Calendar Ye&r i9S9 



Tftble i 



Subcutaaeoas Tusnors in Mice Receivia ^ Repe aled Moofchly laje ctioaat of 

Fi'actia^s of Ait Pollutanls of ESsht Citiees, 72 Mice ia STftch Se?leSc 



Pro P„ Kotia'g D&ta 
Bessol Arezjnatic O^fgem&tdd Aliphatic 
FracUoa Fpactioo Fs-actiosa Fractieta 

Ttsmors NOo Tujysora NOc Tumors No. Tunno?« Tot&l 



Bi^miagham 


4 


Cisicaajj&ti 


1 


Dslg"©it 


§ 


Log Angeles 





Nei» Oxleaas 


i 


PlailBdelpSiia 


3 


Saffi Fraacisc© 


4 



Total 




4 
I 

2 

2 
_ 1 
10 



9 

J 4 
4 
7 
5 
5 
5 

-A 

54 



ii 1 i 







Table 2 












Beagpyrenie Expe^imest 










C=57 Black Mice 












D&te 


Total 


Tctel 






Expo 


Dose 


First 


No. 


lajected 


No, 


Total 


No» 


Mg„ 


lajectioa 


XQJeetio&s 


MC, 


Tmr.ors 


Deaths 


188 


S 0. 00^ 


4=28»58 




0«00 





72 


186 


S 0. 008 


3-25=58 




00008 





63 


185 


S0oOi25 


3»25-S8 




0„0125 





64 


159 


S0»025 


8»22o57 




On 025 


6 


72 


160 


SOoOS 


8=. 22- 57 




0,05 


7 


72 


162 


SO. 10 


8-23=57 




0,10 


16 


72 


162 


S0,2S 


8»23-57 




Q„25 


19 


72 


163 


S0„50 


8-23-57 




0.50 


19 


72 


18? 


MO. 00* 


4» 28=58 


12 


0.00 





72 


165 


MO0OO2 


8-29-57 


12 


0,024 


3 


72 


164 


M0» 004 


8»29-57 


12 


0,048 


? 


72 


166 


MO. 008 


8-29-57 


12 


O0O96 


25 


72 


167 


MO. 02 


9=3=5? 


12 


0,24 


42 


72 


168 


MO0O4 


8-30=57 


12 


0.48 


53 


72 


S = SiBgle Dose 












M » 


Multiple Dose 











* Trlc&prylin Controls 



ai 9 



- 'i • J-sriai No. NCI=2iCibJ 

Calendar Year 1959 
_b__C'COBt| Sigaaific aaee {c&nt) 

ceiidtituecta of this natu^s originati^ f?ozn automobile exhaust &^q 
moBt imp^s'taat. In coM^ast^ the l&igh easicer yield obtaiaed with f^acti&iets 
obtaitaed from Bismit&ghasn, Alabama, was related to the fractions coa^aia^ 
iag a?©ma^ic compouade. This estesepvatioa is in genes-al agreemeol wife 
the fact that ^e pollutants in this eily are nnaialf eoot from coal bu?Eiiag 
furnaseso thai acc^s-diisg to chesnicai analyses znade by the Saeitafy 
£ngin@@:riag Ceate?, the pollutas3.t9 of Birmingham had the highest 
3„ 4='benapf rese conteat of the antiye series, and with the epidemiologic 
data., that ^e lurag cancer death rate amoog inhabitants of 163 xn'etre° 
politaa areas in the United Stales was found to be highest in Birminghanrio 

S^ch a parallelisir. does not extend to some of the findings m&^e^ 
indicati^ hereby ^at additioaal air pollutant factors of different sources 
a§i,d nature a?@ appares^tlf active in determining l®cal lung cancer rates 
ia differei&t c®znzn unities. It is„ for instance^ possible that the high 
ch^esnimn coatsM of Ih® air pellu£an«^3 collected iu Baltimore tsi&f exert 
a d®.Ciail® iaiiueace in this sespecto Doubtlessly, the total lung caacer 
fate i@ iKiidividual cosnmunifies reflects the total and combined action of 
a variable carciaogenie conripleiSo ^hich probably is even znodified by 
the simultaneous presence and actioQ of various noncarcinetge^ic "ce^ 
carcinogens", io eo , respiratory irr£tant8 injuriog the defense mechaadsms 
of the respira^ry mucosa and faciliSaUng and pz>@longing the action of 
atmosphes'ic carcinogens. 

When the results €>btai!^ed in mice are coin pared vd^ the tunrsor 
yield found in the gradated benspyrene series, they are in the rasbge of 
9, 02S to .0, 10 mgmo of 3. 4-beaspyy9ne given by a single Injection or 
in that of Oo QQZ to 0. 006 administered by 12 monthly injections. 

Prop«»sed Courses « " The first phase of this study has been terminated. 
The histologic ^ork remains to be d@ne. The final 
report "s^ill be prepared in cooperation with the Sanitary Engineering 
Center and Dr. Kotin. Studies on total air pollutants intra plsurally 
administered bensol es:tract and inorganic fractions will be coaitinued. 



Part B Included- - - - YES NO 



h^ '^ 



Serial No. NCI=2U^a| 
Ca?.sK!Sa? Year i9S9 

2. Eavirsajmeratai Career 

3o Betlsesd».o Md, 



I&dividi&al Project Report 
Caleadar Yeas- 1959 



Pay* A 



Project Titles = Epid€Siy}i©l@gical Stedies of Eavigozsmeatal 
Cagcino^eas 

ia,) Epidemiological Study of hang Cancer ia Wo^kes-s 
Exposed to the lahalatioa of Ziae GSsromate Paiot 
in Aiffpiaa© Coastrwctioa aad Maiaten^-ssce PlaaSs. 

Principal lavsstigatoi? ^ =• Wm, V^, Pafiae 
Othe? lavee^gator - » = - Yf , Co Huepas- 

Cooperatiag Unites - U„ S. Ai? Fo»ce aad a 

private aisplaae rnaaufeclus'er, 

Mas Yea^s <cal€u?.dar year I9S9I 
Totals ° « I 

Professioaals ° , I 
Others = ,0 

Objsctivees ° » Epidaznioiogical studias h&VQ showa tbat wos^kers isit , 

. ch7onnat®'°°produ€iQg plai&ts have aa abnormallir high 
iacideace of cancer of the lusig. Observations in Eusops suggest ths.t 
workers exposed t& cferomium pigm'eate also experience & respis-atdify 
caeceg' feasag'd^ bat epide?niological eYidaace is Isckiago ^ 

Tlie air'plaae maQufacturiag a&d ntiai%$:eiia3sce isdustrf appeai^e 
to offa? tJas h@Bt oppor^ziity for studying workers who have beea exposed 
t© sitae chromate paiets aad related compounds,,, The Office of the Sui'geoa 
Geaeral and Ihe Air Materiel Commaad of Iha Air Fej?CQ„ aad Iks disfscter 
of Health aad Safetif of a large airplan.© sKaaufacture bave espresssd &n6 
sho^a a williQgsiess to cooperate ia thi^s Btudf which is designed ta 
determine vi^hether expssure to sine chromate psiat by iahaiatioia ifesull;s 
in aa increased risk t4> l»i!,g caacer. 

Method@s ° ° From ^e persoanel rscojrds of t^@ anai^teaa^ee bases asd 

©ae coastructieB plaat the siames and lesgth ol sesrvice 
aad other identMfi&g data will be obtained for all spray paiat«re v/ho are 
ao iofsger employed in tha respective eslablie&meats. Sisrdlar ioforrnatioa 
vdli \na obtained for a group of co^atrolso 

!;.1 b. 



Caieadar Year 195 9 

Th?o^gh Social Security aad Civil Service Retiremes^t Board 
v&CQ?48 of each v^orksr vifill be traced t& ascertain whether he is 
still iivlBLgo II liae per@@3s> has died, his death cer^ieate 'will be 
obtaii&ed aiid the diagoQsis of cause of death will be verified if possible^ 

Major Fiadii^ss - -= Otnlj prellmiaary lavestigatioBS have beeu made 
to dstermiaae whether the project ie feasible« 

Sigeificaaces ° «' Siace dbremaSe paiats are used so exleosively,, the 

results of the studf t?ouid be of general public health 
iateresto The investigatiott will afford aa ©p|K>rts3nity to iacreaae owy 
knowledge regardi&g the earcisogenicitf o£ snetala a^d will zvtppleynem.t 
imf@?zr>atiO!i gained it&tn laboratory eKperimeats. 

Proposed Course of Projecti ■=> ° The first phase of the project caa^ 

be com plated ia about six snaths. 
The remaiaing ^?ork of tracieig the '^vorkera, obtaieisig death certificates 

assd analysis of data will aol require as much issgeosive vs&sk bat v/ill 
exteud over a losger periods 



PART B fecluded?=----YES NO __X 



klE 



:ii 



Ses-ial No, NCIo212 a- 
I, OADR 

3o Bethasda. Md, 



PHS-NiH 
Individual Project Report 
Caleadas- Yea^ 119S9 



Part Ao 



Project Titles - ° Sagpegimeatal Cagcigiogeaeais ef £h© Upper 
Digeative TgacI 

Piriacipal lavestigaloy » » M, Ku?atsuae 
Other lavesfegatos- » » » W. C. Hueper 

1, lavestlgatioa of "Seis-ogaa" 

OlJjectivess =- in Japaa, as iS is xwidely kao^a, aa excep^eaallf high 

xnorlslity r&ie &f gasts'ic cas&ceir has beea srecordado 
Maray investigators have tried to clarify fee causes of eucfe a usiiqus 
phaaomeaojas, but an^ acceptable eKpianati®a d®®s not seem &c» have 
baes smppUed yet, "Sei?ogan", a p&6ffiES mediciae ef Japan, seems 
iate^estis^ in this respect because it consists <c»f maiaif e?@oso$e ai&d 
has bees mdely used for Kiore Ihsn 50 yesye in Japsau The preasjat 
study is t© deteemiae wheOaer ®r aot the ixiediciae is caffcinegeoic 
foar experijTieiital aiiimals aad if s@, what ceznpoaieQl or coimp<» treats 
miB'e r«epoi%@ible for ittc 

Metheda Used? - - A eainple ®f "Sesifsgan" -was collected fsrom a d^ug 

at&s® its JapaOff It is sstnali Mack- colored pills with 
a vexf Bt^eag ©dor characteristic of creosote. Ite iaga-ediemtss are 
gly-cerine„ medicinal creosote,, aad aom® ether matesfiai fyom plant 
8®urceffi, The sampl© coUscfed -was exfes'acted with dicMoPomethsne 
aad the extract was uasd fer ^e follawisg ea;pes'i3ri©sats<. 

&)) Prelisniaa&'y chemical aoalfsiSo 

A small portien of the esctsact was subjected t® a series of 
chsromat^grapMc f?acfei©aati®ne and the f^actioaa thus ofeeaiiied ^ere 
«%amiaQd speclrographically. 

b| Subcutaneous i&jection o£ the extract. 

Fos-tf five male mice of C5? BL/6JN strain were injected with 
Oo OS mlo ©f 10% solu^oa ©f "Seirogan" extract is fcE-icappyliae at fee 
iatrascapuia? region forlmghtlyo 

.Mam ^ears Ccalendar je&r 1959) 

TM® project iacXuded with NCI^Zli - Total, Professional, Othes' 



da? Year 1959 

c) Paiia^isg ©f the ©xtFSCto 

Fortf five male mice ©f C57 BL/6JN et?aSffi «rerc painted 
wish the exts-acl ©a the back 3 times & weekc 

Res«it®g - The espefimeR^s ware ©aly receatlf started. The results 

BH far obtained aire as follows s 

a] The preliminary chemical aimly@is showed that the exSr&cl ceaaiste 
maiidf ckC ph@a@l derivatives and thei; oxidation productSo No 
detectaM® amount of feaaae^alpf reae aad ether knowa asomatic 

carcit^geat ■was fouado 

h] Svibcnt&n@&%is iajeetie^ of the extracto No worthwhile results are 

a® jst available. 

c] PsiEtissg of the eactracto P&iatissg Ms beeai pes^formed fosr abouil 2 znoai&s, 
TMckeniag of the skin aad fo^xnatioa of small aodules in the sub» 

cutaa<9ous tissue at the aite of applicafioa <we?@ @bse?ved. 

Sigaif leasee @s ^ <> Sisace creosote is used as a heme remedy not oi^ltf ia 

Japan but also ia snoat Asian countries, and its habitual 
usage is a common practice fe? some Jap&aeee, the results of tWi preseut 
etudf m&j be very isifornnali^e iot carcloogeuesis @i the upper alizneata^y 
tracto 

Proposed Course; ° » Feedistg of the extract will be undertakeisi in order 

to clarify its possible effects €>a the digestive &zgti.u9o 
The effect of tho extract Gn the mucosa of the giaadular stomach i;dU be 
mvestigated by i&jdcti«);§ the estraci it&t(» the mucos&c A more detailed 
eheznical analysis is also contemplated, Epidemiological re-e^camiaatioss 
of the esophagus ac&d gastric eaiK:er ira terms of consumption of "Seirogaa" 
aud other creosote mediciQ&l preparatioas in. Japau are uade? coosideratioo.. 

2o Mimmum effective dose of beesoCalpyreae and dibeazta,h) anthracei^e 
to produce carciaoma ie the glandular stomach of mice, 

ObjectlTees ° ° The previous iuvestigatioae of the preseat authors and 

a«%d other i&vsstigatore have demonstrated the presence 
of many eictrinsic sources of known aromatic carcinogens which can 
daily supply &ose carcinogens itato the human digestive tract. The 
magnitudes of 6he carciK@gene ^us ingested seems te be rather small^ 
however. Therefore, it seems interesting to determine whether such 
a small amount of the carcinogens can actually induce carcinoma in the 
glandular stomach« 'when introduced into the submueoea., Among tha 



^17 



Serial No= NCI-212U and 2^ 
Caleadajf Year 1959 



ECco©.t| 



carcinogens bec^sofalpf ireae ^as cbs-sen^ for tlie p^'eceni inveaetigatioae 
because 11 is the most c@mmoQ and most potest carcinogea as^d has Rtot 
as ^"et beea in^estigaftsd im terzna ©£ iatramural iajeclioffi. DibeBSsCaofel 
anthS'aees&e v^ae als® selected because the isitramu^al admitdslration of 
its tsrac® ameuct bas aot yet heew. itsvmaUg&ted^ 

Methods Smpioyeds » » Beas©<a|pyreiiie aad dibeQzCa,h)aalhr&ceae 

have been purified by repealed eh:romatog7aphic 
f^actiosm^oaeo Dibei^s^aathraceae was purifisd to colorless crystaICo 
Tbe purified ca?cii%dgs!&8 xnewe dissolved in tricapylia <which had been 
pafftially purified by shakiag '^&i aclivatsd carbOQo Three different 
solutiosss of beo^pys-ese &ud tt»o aoluiioi&s of dibecs^anthraceae ^ere thus 
prepared &n4 0o0C171 mL @£ each e@lutien injected into'^e gaslTic glaadulai 
muc&sa c£ each irtouse hf using a calibrated mlcro^pipet ainched lo a 
30«gauge hypctdermic ^eedlco The solutions injected eften leaked c»ul of 
the iajectiaa sitSo They ^ere carefully absorbed @a a strip of filter 
paper mider ultraviolet radialio^&g eluted wiih isooetasee then the exact 
sumount @f the carcitiegea which had leaked out was deterxniaed spectre^ 
graphically. Thus, the amotsi&l el beaspyreae or dibenz°anthracefie retained 
in th.0 gastric ^all was accurately calculatedo (The calculatioaa , hov^ever, 
have 920t yet beeo completed).. The amouot of carcinegens injected and 
the number of aaimais ueed ass&s f&llowss 

Control 
Tricap" 

Bea apyreae Dibeazanthraceae rylin 

^^ 16„ inracg, l. hlmcg, 0. I6mcg» 6. 28mcg. 0, 67 meg. 0. 0017 mL 

Noo 

of 

Mice 38 41 46 40 43 41 

Eetsultss ° ° No results are yet available since the eKperimeats have only 
recently beea started. 

Sigaificamces » . If carcinomas ia the glandular gastric mucoea should not 
i^ elicited by these procedures, it is difficult to c(»aceive 
that the two carciaogene used play a& important role ia the caus&tios of 
cancers of the human tstemach since they would show that even on direct 
contact of the glandular mucosa this tissue is refractory to the carcieogenic 
actio!& of tiiese chemicals. If, on the other hando a significant number e>f 
carcinoznas is induced by this method, evidence is obt&iaed for undertaking 
the second phase of this in^'estigaUoa and it will then be attempted t@ create 
proper experimental conditions enablif^ the penetration of similarly nninute 
amounts of these carcioogens to penetrate the "muceus barrier" and to 



418 



ifcoat) Sigsajgi eaac® (c© at)) t 

■^tit^v the gias^dukis' mucosa aud submucosa of th« atenmach whem th€:ee 
c&^cioogeeiS £?e givsa css?aUy<. Tbe ppeaeat »feidy„ therefore:, may supply^ 
fusd&mesital i'M^^maMoii of dis'ecl practical sigmfica^es %e t@ the poseibic 
ir©l® "ffi/hJcli c.&?<8ia©g®E5ic as^sn&tic fey«lr©carbo»a ingested by feumaas with 
theiff food isi&'f pl&f in the pFeductioD csl gastric caacero 

Ppospee^i-vQ Stediee? =*la a secoad sasiea ©f aaiKjale repeated i8i«:ramttra2 

iQJecti«»@.s of similar aniouMs of the listed ca?eiao- 
■i^ns -^Ul b@ m£d@ fo?f testisg the csas-eisogeoic effect ef these caircittogoss 
'?.--h.ea acMag ©'vsr a ioag pes-i^d of t-im e, 



Part B lueludeds =■-■ - YES NO X 



li"! Q 



Seri*l NOo NCI=2i2C3) 
- 5 - Calendar Year 1959 

3. P@aetrsti©ia of fe«aaoia|pyrese iiato the gastric glandular mucesBo 

OI>j8elivess = = la o?day to facilitate cas'einogeoic palfc^^lic aroma^c 
S&fdroear&oncs te penetrate the glaadular mucosa o£ the 
»tt3>tn&cho it has feeea a common practice t@ empley lipophilic a@ well as 
h^dropfeilic m&tesiais as a vehicle of the carciaoge&«o It seer^ed 
istesastiag fes ka©<iw tphelihe? aay sigaificaat amount o£ &e cayciaogeas 
p®?02raiiy adunlBislered cae peiaelyate the gastric glandular mucosa whea 
dissolved la diluted ethaoel. aqueous caffeine solution„ or a mixture @f 
thes© not ealy foscsus® these selveafes, though h^drophilic. easily dissolve 
the csis^ciuogeas^ but also because they are frequently inlreduced into 
th9 h.ikssi&x& digestive canal. 

Me^ods Emp'ioyeds =■ = A% first, the solubility of benzoCa^pyreno in 

aqueous ethanol solutions, caffeine solutions, and 
caffeine -etha££Ol solu^o^s, was determii&ed by shaking crystals of the 
carcinogen in these solvesite at 23®C for 12»l§ hourso Some of the results 
of the determination ^sre shown as foUowss 

Solvent Systems n„ 1ft o= 1% 0,1% 

Caffeine Caffeine Caffeine 

10% 20% 30% 0„1%^ cent 10%conto20% c©nto30% 

Ethaaol Sthaaol Sthanol Caff eine ethanol ethanol ethaool 

Atnt, of 
BenaoCaJ^^.^^^ ■; 



disscslred e.,010 0., 116 Oo 17 Oo 24 0.38 0„ 98 

(nfscg. /ml) 



X 



The average caffeine coticentration of tea and coffee is approx., Oo l%o 

TliuSo it vf&a found that caffeine methanol solutions dissolved much 
more benspyrene than single caffeine or ethanol selutionSc Therefore, 
in the following experiments « the benzpyrene saturated solution in Oc 1% 
aqueous caff eiise selution containing 30% e^anol was mainly used. 

The solution v/a.e introduced by a gastric tubing into &e stomach 
of mice <^hi£h had fasted for about a dayo The stomach ^»as delivered 
30 minutes, 1 houre or 24 l^urs after the a&ninistration and its frosen 
sections were exan^iined under a fluorescence microscopeo 

Resultss ^ " The main results so far obtained are that the mucosa of the 

foreetomach showed a very strong fluorescence of beB3pyrene„ 
v?hile the mucosa of the glandular stomach « an extremely weak one. Thus, 
it seems that only trace amounts cf benspyrene penetrate &e glandular 
mucosa ev©B ■whsE it is dissolved in the present solvent mixture. It may 



ii9n 



Caleadat? t^&r 1959 
^ 6 =^ 



Reisults (coat) 



be possible that even in this f^rzn of administfatioR & streng competitive 
ediitxity of the ^mueosa of the forestomadb attracts the mais^ poz'tiOQ. of 
the betispfs&ns int?@ducedo 

Significances ^ Siiaco both ethsii&ol ^od caffeine are cozninoely ingeated 

by maa, th® preaeat study may supply practic&I and basic 
iaf^rmatioa ia regard t© human gastric csrciaogeaesiSa whea it ia complete 

Proposed Courses = «• it is plaaaedt© determiae -whether or ao^ repeated 
oral admimstrati©® of beiaspyreae dissolved in the 
caffeiae methanol c&a penetrate th& gastric glandular mucosao 

It seems iatsreeliag to kso^ if betxspyreae caa eater Oie glasidular 
mucosa of the rabbit because this aoimalo like mau, d&ee not have ^he 
foreetomaeho 



Part B laciudeds YES HO 



Serial Ifoo NCI-301 

is Labo of Blocbem, 



C.O 



Office of fcbe Chief 



3o Bothesda/ Mdo 



Part A, 



Project Titles S«scx'etsry Duties in the Lab©r>atos?3r 
©f Biocheuiistpy. 

Principal In-y©stigators: A, E, Stotler and 

Jo Ko Shores 

Other Investigators: Hone 

Cooperating Units: Hone 

Man Years (calendar year 1959): 
Total; 2 
Professional: 
Other: 2 

Project Description: 

O bjective s: The objectives of the Secretarial Unit ' 
In th© Ofi'ice of the Chief arrs to: Maintain psrsonnsl. 
correspondencSp reprint and requisition filesj 
maintain leave^ overtim©^ and budget records, xnairjtaiD 
a Laboratory library^ arrange for travel of 
professional staffs compose and type letters^ 
memoranda, requisitions^ and shop orders^ and type 
manuscripts J, lantern slides^ travel requests^ 
personnel requests^ and miscellaneous charts and 
articles a The above duties are performed for 
36 professional and 22 subprofessional employees o 

Methods: Since January 1959j this Unit has typed 
IShe following: ApproXe 3000 pages original rasnuscriptj 
1^80 letters and memoranda, 50 lantern slides^ l|.00 
pages of miscellaneous material, l500 requiaitioss^ 
175 instrtiment shop orderSj^ 175 Request for SsrviceSj 
50 reports of surplus proper ty^ 25 Shipment Requests, 
60 Travel Requests, and ij.0 Personnel Action Rsqiiestso 

In the future this Unit will continue to do its utjaost 
to produce as much work as possible end to perform 
this work accurately and rapldlyo 

Part B included « No* 



42 



9 



Serial NOe NCI-302 
PHS-NIH 1„ Lab o of Biochemistry 

individual Project Report 2o Office of the Chief 

Calendar Year 1959 3o Bethesda, Md, 



Part A; 



Project Title; Microchemical empirical analysis; 

Development of micro chemical methods? 
Infrared and ultraviolet absorption 
spectroacopyo 

Principal Investigators: Robert J, Koegel 

Other Investigators: Kathryn Baylouny, Jasjes Williams 

Dennis Clark 

Man Years - (Calendar year 1959): 
Total: 3-I/I4. 
Professional: 1-1^ 
Other: 2 

Project Description : 

The microchemical unit provides a versatile analytical 
service in collaboration with the independent investi« 
gators of the Biochemistry Laboratory by applying the 
most recent developments in analytical chemistry to 
their particular problems and by developing new techni- 
ques as their problems requires 

The routine service of the unit continues as described 
in previous annual reports: however, during 1959 
some 5^500 samples were submitted for ana].ysi3« IMs 
figure when compared to the corresponding one for 1956 
reflects a two-fold increase in the number of samples 
submittedo This increased ^^orlc load « an approximate 
increase of fifty percent - was adequately metj, although 
the unit suffered a decrease in total man years of sosie 
15 percsnto The work year of 1959 has, therefore, been 
th® most productive in the laboratory's history o 

The solid state infrared absorption spectra of some 
twenty percent of the samples submitted for empirical 
analysis are now being routinely run and the recorded 
spectra made available to the individual investigator 
for th© purposes of structural interpretation and as 
indicators of chemical purityo 

SSE CONTINUATION SH33T (A) 

Part B included Ho 

423 



Sorial No* KCI-302 
COHTINUATION SH3d3T (A) 

The infrared research program as described in the annual 
reports of 195? and 1958 was continued on a very reduced 
scale during 1959o It is planned to prosecute this program 
rigorously during i960, extending the studies of the optically 
active amino«aci(3s and peptides into the 15[j. to 30^1 region 
amd also to examine the influence of adjacent functional 
groups on the sioleuular absorption coefficient of the 
carbonyl stretching motion of the peptide structurea 



421^ 



Serial Noe jlfCI-306 
PHS-UIH lo Labo of Biochemistry 

Individual Project Report 2o Physo Chomp and Chroma= 

Calendar Year 1959 tography Section 

3o Bsthssda, Mdo 



Part A. 



Project Titles Studies on Hydroxyaznino Acids 

Principal Investigators? To T« Otanip No Izumiya^ 

Mo IfinitSj Se Mo Birnbaum 
and Jo Po Green stein (deceased) 
S« Akabori. 

Other Investigators! None 

Cooperating Units! None 

Man Years (calendar year 1959)! 
Total: 3ol}. 
Professional! 2ok 
Other: 1 

Project Description: 

Treatmejat of an alkaline solution ©f an smino acid 
with aa aldehyde^ in &e presence of catalytic amounts 
-of cupric -ionp proceeds with th® f enaation of the per- 
tinent a«»amino<=»6hydroxy acid according to the folXoH<=- 
ing scheme: 

ECHO + IJHgCHR'COgH ^^^^L^ RCH(OH)CR»C5IH2)CO H 

Such reaction scheme penaitted the single step y>^®'^ 
paration of 

(a) fS-^hydroxy^p^^methylaapartic acid from pyruvic 
"^ acid and glycine j, 

(b) p<=>hydr0xyaspartic acid from glyoxylic acid and 



(e) p-hydrosyleuein® from isobutyr aldehyde and 
°" glycine 

SEE CONTINUATION SHEET (A) 
Part B included Yea 



i|25 



Sarlal NOo NCI».306 
COHTIMJATION SH3ET (A) 

(d) a-sthylsarine and a-methyl serine from formalda- 
hyde with BL«a-»ainino-n«butyrlc acid and PL- alanine 
raspoctively, and °^** 

(e) serine from formaldehyde and glycine* 

As p=hydroxyaspartic acid, p-hydroxyleucin© and |3-hydroxy« 
p-methylaspartic acid contain two asynsmetric carbon atoms , 
these were presumably obtained as mixtures of the two 
diastereomeric racemates. All of the products (a) exhibited 
single spots after paper chromatography in four "different 
solvent systems, (b; revealed elemental analytical values 
in agreement with the proposed structure and (c) could be 
reduced to the corresponding non-hydroxylatod a-amino acid 
by the action of a red phosphorus-hydriodlc acid mixtureo 
In additlonp p-hydroxy leucine released the expected one and 
p-hydroxyaspar^ic acid (smd the p-methyl analog) the expected 
two moles of carbon dioxide upon analysis by the manometric 
ninhydrin~C02 procedureo These results, together with 
previously described syntheses , suggest that the method 
Is a general one and provides both a simple and an econoiaical 
route to a- ami no- p-hydroxy acids o A number of such hitherto 
unavailable a«amino-p«=hydroxy acids are presently being 
prepared o ^ 

A new synthesis and resolution of 5»hydroxy lysine , and the 
configuration of the 5-asymmetrlc center of hydroxy lysine^ 
which has not been hitherto established^ is presently under 
investigationo 

Significance; Availability of a-amino-p-hydroxy acids of 
various structures should permit us to make use of these 
materials as enzyme substrates and henc© help enhance oar 
knowledge of the mechanism of action of biological systomSo 

Proposed course of investigation; To study the effect of 
a-aminO" p-hydroxy acids so prepared as anti-metabolites 
and to investigate their susceptibility to various ensyme 
systems such as amino acid oxidases, decarboxylases, serine 
and threonine dehydrase and transaminases « 



42B 



Serial U©o KCI°306 



PHS^SJIH 

Individual Project Report 

Calendar Year 1959 



Part Bo Honorsj Awards^ and Publications 

Publications ether than abstracts from this project; 

Akaborij, So^ Otani^ T, Top Marshall^, Res Winitz, Mo, 
and Greenstein^ Jo PoS A Synthesis and Resolution 
of DL=S®rln©o Archo BlocheiQo Biophysop Q3j, 1-9 
(19^) o -^ 



427 



i 



PHS-UIH 
Individual Project Rspox't 
Calendar Year 19^9 



, Koo KG 1-313 
Labo of Biocheraistry 
Physo GheBio and Chroma- 
tography Section 
B©thesda, Mde 



Part A, 



Project Title: Chromatographic Analysis of Protein 

Components of Serum in Health and Disease, 

Principal Investigators s H„ Ao Sober and E, A, Peterson 

Other Investigator: H« B, Levy 

Cooperating units: NIAID 

Man Years (calendar year 1959) 
Total ; 1^2/3 
Professional: 2/3 
Other: 1 

Project Description: 

Objectives: To develop and apply chromatographic pro^ 
cedure's' t© the isolation and identification of the many 
macromolecular components in norraal and pathological 
serum and plasmao 

Methods employed; Plasma^ serum, and their preparations 
are subi'jecte!! to chromatographic fractionation on cellu^ 
losic ion exchangers^ using gradient and stepwise elu« 
tion techniquasp The resulting fractions are essminad 
fey physical^ chemical ^ and iirniunoshemical techniques, 
and specific biological activities of these fractions 
are eharacterizedo 



1(> Further study of the completing of human ssruai 
mercaptalbumin with lipids has shoi-ra that whereas the 
addition of a fatty acid markedly increases the affinity 
of the protein for DEAE<=cellulose^ the addition of a 
cationic lipid (eetyltrimethylarsmoniua bromide) can 



SEE CONTINUATION SHEET (A) 



Part B included 



Yes 



Li28 



Serial Noo Mci-313 

CONTINUATION SHEET (A) 

reduce its affinity for this adsorbent to zero. Evi- 
dently these substances ar© bound to the protein through 
their hydrocarbon chains. The complex x^ith the cationic 
lipid is adsorbed more tightly than >t«globulin to CM- 
cellulosej, indicating that it has becnrae a highly basic 
protein, 

2o Simple rapid procedures for the preparation of over 
90^ of the ''^-globulins of serum^ free ef the macroglobu« 
linSj but still containing all of the usual antibody 
activity (10 antibodies were tested) have been described 
and with Dr* Levy of NIAID have been adapted for use 
by imraunelogists for the preparation of high titer 
antibody serap- Macroglobulins, relatively free of the 
smaller 7S rf^globulin, can be prepared by zone electro- 
phoresis of" the remaining protein, 

3« A variety of metals (calcium^ barixaa^ strontiina, 
chromiumj, magnesium^ as well as iron and copper) have 
been found^ in collaboration with Dra B, Vallee of 
Peter Bent Brigham Hospital^ Boston p Mass,, ^ to be 
localized in discrete fractions of the proteins of 
serumo In some cases several chromatographically 
distinct fractions containing the same metal have been 
observedo 



Ic Changes in the chromatographic behavior of the serum 
proteins must be well understood if the chromatographic 
patterns are to be interpreted with confidence o Albumin 
is the major protein component in serum^ and it appears 
to be the most susceptible to alterations of this kind^ 
so study of these effects in albumin are particularly 
important q Infoimetion already acquired has shed light 
on the nature of the heterogeneity of albtsicln as observed 
in seriam chroma tegrams^ which may reflect its transport 
function, 

2o Purified antibody fractions are required for fluores- 
cent antibody studies^ for sequence and structure deter- 
mination s^ for reagents in typing procedures and for 
clinical use as potential prophylactic or therapeutic 
agents 9 

SEE CONTINUATION SHEET (B) 
Part B included Yes 



ij,9Q} 



Serial lloa HCI»313 
CONTINUATION SHEET (B) 



3o The existence of trace inetals In biological sys- 
tems has long been recognized but their structure and 
function hmiS not yet been elueidatedo The recognition 
and purification of inetalloproteins should pei^it such 
studieso It is assiamed that the metal bears some inti- 
mate relationship to the biological activity and thus 
provides en internal label for the r^^ive site* 

Proposed course of the project ; 

la ' The complexes obtained by combining albumin with 
anionic and cationic lipids will be further characterizedo 
Complexes with nonionic lipids and detergents will also 
be studiedo An attempt will be made to prepare albumin 
free of all lipldo 

2o Studies designed to purify and characterise the raacro<= 
globulin components of serum are being continued with the 
purpose of defining the subunit structure and the 6ggre.»> 
gation mechsnismo 

3« Isolation^ purification and characterization of the 
metalloproteins has just begun. It is hoped that with 
the purified compounds^ unique structural and biochemi- 
cal properties will be discoveredo 



i|30 



Serial Noo WGI»313 

PHS-NIH 

Individual Project Report 

Calendar Year 1959 

Part B o Honors^ Awards^ and Publications 

Publications other than abstracts from this project: 

lo PetersoHj, Eo A.p and Sober, Ho A*: Chromatography 
of the serum proteins « Plasma Proteins, edited 
by Po Wo Putnamo Academic Press, New Yorko 
In press, I96O0 

2o Levy, Ho Bo and Sbber^ Ho A«: A simple chromato- 
graphic method for the preparation of gamma glo- 
bulino Proco Expo Blolo Medo In press, 1960o 

3o Sober, Ho Ao and Peterson, E. Ao: Protein chromato« 
graphy on ion-^exchange cellulose o Fedo ProCo 12_: 
1116-1126, 1959o This paper was listed as "in 
press" in the 1958 reperto 



431 



Serial Noo NGI«335 
PES-NIH lo Lab* of Biocheraistpy 

Indi'ffidual Project Repoi?t 2o Physo Cherno and Chroma- 

Calendar Year 19^9 tographj Soetion 

3o Bethesda* Mdo 



Part Ao 



Project Title; Quantitative Nutritional Studies 
with ViJat ©resoluble 9 Chemically- 
Defined Diets 

Principal Investigators: M^ Minits» So Mo Birnbaum* 
J^ Po Greenatein (deceased) » M. 0© 
Oteyg, To Sugimura (visiting scientist) 
and Vo Mitbander (visiting scientist) 

Other Investigators: None 

Cooperating Units: None 

Man "Years (Calendar Year 1959): 
Total: 9 
Professional: 3 
Other: 3 

Project Description: 

The formulation of a nutritionally adequate^ 
chemically defined* crystal clear liquid diet 
composed of l8 crystalline L-anino acids (except 
for glycine )» the water-soluble vitamins* the 
pertinent salts* glucose? the fat-soluble vitamins 
and ethyl linoleate as the source of essential fatty 
acidn has been achievedo Such liquid diet is pre- 
pared as a transparent » clear » stable emulsion of 
the latter two fat-soluble ingredients in a $0% 
solution of the other ingredients in water with 
the aid of a small amount of polyoxyethylen© (20) 
sorbitan monooleate (Tween 80)o The lipid phase 
reveals no tendency to separate or become ranoid^ 
and the clear diet may be stored at room temperature 
or at <^20o for prolonged periods of time without 
apparent deteriorationo Weanling rats fed these 
diets ad libitum for a period of over 3 months 
exhibited no signs of essential fatty acid deficiency 
and* indeed* grew at the same rate as did controls 
that were provided the fat^soluble vitamins in 
Ma^ola (com) oil as a supplement© 

Part B included: Yes 

SEE CONTINUATION SHEET (A) 

432 



Serial No« UCI-335 

CONTINUATION SHEET (A) 

The majos? portion of the studies presently being 
carried out represent an extension of the previous work 
with water-soluble, ch®aically=defined diets o Previous 
studies with laboratory animals revealed that such diets, 
composed of crystalline essential and non-essential L« 
amino acids, vitamins^, the required salts, glucose and 
the essential fats, may be satisfactorily employed in 
long teiTO nutritional studies, including those bearing 
©n longevity, reproduction, lactation and various 
pathologic conditions o Unique features of the diet 
include 2,t3 chemically defined cmapositien and its 
extreme water solubility (it is generally administered 
as a S0% solution in water) and thus permits nutritional 
studies to be effected with a high degree of quantitative 
precision and with diets wherein one component or a grovsp 
of components can be added, deleted or changed at willo 
In recent collaborative studies with otjs» clinical associates, 
the complete diet has proved most adequate as a source of 
food for patientSo At the present time, the chemically^ 
defined diet is being employed in the laboratory to study 
enzymes and metabolic processes in vivo^o 

Sjjggif icance : Certain features possessed by water»solubl©, 
ohaaicaliy (defined diets make them of potential practical 
application in a niaraber of ways© Such applications 5.nclude 
(ajuse for quantitative metabolic studies in man and 2n 
laboratory animals, (b) food for germ free animals, ic) 
food supply in space travel, Cd) treatment of patients" 
with metabolic dysfunctions, C^) food for patients with 
ulcerative colitis, (f ) pareht'eral feeding, ig) treatment 
of kwashiorkor and ChJ use for study of specific enzyme 
systems and metabolic" processes in vivoo'Thus, our own 
use of such diets for qussjtitsstivi" F^itifeolic studies in 
laboratory animals has permitted the accumulation of data 
on nitrogen balance* and nitrogen metabolism with a degree 
of precision and certainty not hitherto achlevedo Comparable 
quantitative metabolic studies in man, using these same 
diets, are presently being carried out by our clinical 
associateso As a potential source of food for germ free 
animals, these diets possess a distinct advantage in that 
the whole diet can be sterilized by filtration, or the 
separate components ©an b© sterilized by filtration or by 

SEE CONTINUATION SHEET (B) 



ii33 



Serial Noo NCI-335 
COHTIWUATION SHEET (B) 

autoclavlag and subsequently admised under sterile con- 
ditions o As a potential food supply in space travel^ 
chemically defined diets afford complete and adequate 
nutriment in highly concentrated formo This is strikingly 
revealed by the fact that only one cubic foot of the dietp 
as a $0% solution in water. Is sufficient to provide a 
man with the required amino aclds^ vitamins, salts and 
essential fatty acids, in addition to 2000 calories per 
day, for a period of nearly a montho Per the study and 
treatment of patients who suffer from kwashiorkor or from 
certain metabolic dysfunctions, such as oligophrenia 
phenylpyruvica, the diet possesses an advantage in that 
any component or groi:?) of components can be added to op 
eliminated from the diet at will. As the diet is rapidly 
absorbed In the upper porticm of th® gastrointestinal tract 
and contains no added bulk, it should al.so prove of value 
for thd treatment of patients who suffer from ulcerative 
colitlso 

Proposed course of Investigation t To study, by specific 
alteration of one or more eoitiponents of the diet, the 
in vivo action of certain enzyme systems such as D°>amlno 
acid oxidase, transaminases, the I^ebe cycle enzymes, 
hlstldase, tryptophan, peroxidase, arglnase, ©tce, and thus 
be able to gain a fuller understanding of metabolic processes 
as they apply to the whole organlsmo As illustrative of this 
point, it has already been demonstrated through the use of 
chemically defined diets and the inclusion of specific 
enzyme inhibitors in these diets, that the conversion of the 
D-form of an essential amino acid to its corresponding L-form 
fn the rat is due, at least in part, to the oxidative deamina» 
tlon of the p«amino acid by D-amino acid oxidase followed by 
a transamina'^lon of the a^ke^fo acid so derived to the 
corresponding L»@mino acido It has also been shown that the 
inhibitory actTon of sodium benzoate on D-amino acid oxidase 
can be completely reversed by the addition feo the diet of 
glycine, which is not otherwise provided by the whole organism 
at a rat© sufficiently fast to perait the excretion of the 
benzoate as hlppurie acido This study will shortly be extended 
to the effect of D«.araino acids in the diet on growth and 
physiological processes in tumor-bearing rats, the D^amino acid 
oxidase of whose livers «ind kidneys show a tendency to be 
abnormally lowo 



ii?.h 



I 



Serial No« NC 1-335 
Addendum to Project Report 

Results: Recent experiments with chemically defined diets 
have revealed that they provide a potentially valuable tool for 
the study of specific ensynes and metabolic processes in vivoo 
'■^-hus* earlier workers have shown that certain of the essential 
L>=araino acids 9 such as arglnlnes methionine tryptophan and 
leucine » may be replaced in whole or in pijjrt by their corresponding 
g-isoraers in the diet of the rate Some of our own results with 
chemically defined diets reaffirms these early observations.. 
In this cased the standard diet contained the ten essential 
L-amino acids » glucose snd the pertinent salts » vitamins and 
essential fats* in addition to a mixture of L»proline» ^^tji-^oslnfi 
ethyl ester* HCl;(i raonosodium ^glutamate and E=alanine as the 
source of non<=essenti€il nitrogeno The growth curves revealed 
that when g=ra©thionin® was employed instead of Xi'^methionlne in 
such diets » no change in growth response of weanling rats was 
observed over a 10 day periodo However*) D«tryptophan was only 
partially effective in replacing its corrf spending L«antipode» 

and D^leucine even less effective 9 since this latter amino acid 
had to be added at a level equivalent to loij. times its L°antipodo 
before a significant growth response was observedo Evidently? the 
ability of the organism to utilize a given essential amino acid 
as its g«antipod© varies considerablya In any event* it is 
generally accepted that the utilisation of the 2=araino acid results 
from its conversion to the corresponding L-isomer according to the 
following scheme: 

D-Axoino acid =-«-=^ a-keto acid -^^^ L-araino acid 



Thuss the Diamine acid is presumably first converted to its 
corresponding a»keto acid analog by the action of Diamine acid 
oxidase* and the a=keto acid* in turn* transformed to the 
corresponding L=»amino acid by a process of transaminationo 

As the amounts°"of these enzyme systems are evidently present in 
the organism in limiting amounts a it became of interest to 
ascertain whether* in feeding experiments with synthetic diets* 
these ensyme systems could be provided with more substrate than 
they could effectively handleo Thus* four diets were prepared 

SEE ADDENDUM (2) 



43.^ 



Serial No<. NCI»335 
» 2 " 

which were identical in all respects but for the exception that 
in the first » alanine and methionine were provided as their 
^»isoiaers» in the second© alanine was provided as the L^isoraer 

and methionine as the D^isomer^in the third alanine was provided 

as the D<=isoraer and raethionine as the L^isoaers and in the fourth^ 

both amino acids were provided as their D»formso The growth curves 
on these diets over a 15 d&j period revelled that in the first 
three diets « that is 9 those containing at least one L»form 

of either methionine or alanine « the growth response was very 
nearly the saraeo On the other hands the growth response with 
the diets containing the two g^^isoraers was very markedly diminishedo 
It therefore appeared* in this latter case» that the D«araino 

acids were provided in an simount which was in excess of the 
ability of the organism to effectively convert them to their 
corresponding L=isoiBers commensurate with the needs for increased 
growtho Now if was previously established by other investigators 
that sodium benzoate is a potent inhibitor of D»amino acid oxidase 
^^ v^troo If sodium benzoate would also behave as an inhibitor 
oF g»'amino acid oxidase in the whole organismo then a means would 
be at hand to pinpoint the rate limiting step in the in vivo 
conversion of a g«araino acid to its corresponding L«isom@ro 
Four chemically defined diets were therefore formulated which were 
identical in every respect with the exceptions that the first diet 
contained ^-methionine as the source of this essential amino acid 
in addition to 1% sodium benzoate^ th© second diet was identical 
with the first but contained L=methionine instead of the B=isoia©rs 

the third diet likewise contained L»methlonine as the source of 

this essential amino acid but* in addition* contained sodium 
benzoate in 2F/o concentration^ and the last diet was identical 
with the third but contained D-methionine instead of the £«iscm©ro 

The growth curves on each of these diets over a 10 day period 
revealed that groups on all diets displayed comparable growth 
except those receiving th© diet containing fi«methionlne plus 2^ 
sodium benzoate* in which case growth was strikingly diminished^ 
Such data thereupon pointed to the oxidative deamination step 
as the rate-limiting on©o Comparable feeding experiments are 
presently being carried out with the a^keto analog of methionine* 
which should undergo transamination directly to L-raethionine^ 



SEE ADDENDUM (3) 



436 



Se3?lal Noo NC 1=335 



thereby by-passing the need for the oxidative dearaination stepo 

One further observation is worthy of noteo It will be 
recalled that earlier workers established that th© detoxifiea^ 
tion of sodium benzoate in the animal organism takes place as 
a result of the condensation of this compound with glycine to 
form hippuric acids which may then be eliminated in the urine 
as hippuric acido Thus« if a means could be found to accelerate 
the conversion of sodium benzoate to hippuric acid in the 
whole organisms then it might be possible to more effectively 
reverse the inhibition of Diamine acid oxidase due to ths 

presence of large amounts of sodium benzoate in the dieto 
On the assumption that glycine was not provided by the body 
with sufficient rapidity to detoxify very large siraounta of 
sodium benzoate e diets were prepared of the same constitution 
as those previously described but in which a portion of th® 
non°essential amino acid complement was replaced by glycine© 
Indeed* the inhibition of D=amlno acid oxidase by th® sodium 

benzoate was completely reversed* as was attested to by the fact 
that the diet which contained the D»methioaine and the 2% sodium 
benzoate now supported the growth of weanling rats equally as 
well as did the other three dietso 



^37 



Serial Noo NC 1=335 

PHS«NIH 

Indi-^idual Project Report 

Calendar liear 1959 

Part Bo Honors s Awards » and Publications 
Publications other than abstracts from this project 2 

1<, Bimbaurna S* M. « Gresnstein© M« E« , Winits* Ho and 

Gr©©nstein» Jo P«, : Quantitative nutritional studies 
with water-soluble » chemically defined diets* VIo 
Growth studies on mieeo Archo Blochemo Biophyso : 
78» 2i}.5«2l}.7j 19580 This was in press at time of 
T?58 annual reports « 

2o Suginmra* To » Birnbaum* So M<» » Winitse Mo? and 

Greenisteine Jo Po : Quantitative nutritional studies 
with water^'soluble* chemically defined dietso VII* 
Mitrogaa balance in normal end tumor-b©aring rats 
following forced feedings Arch* Biochemo Biophyso : 
01^ h39-khl^ 1959o 

3o Sugimura?. To » Birnbauia* So Mo » Winitza Mo and Greenstein^, 
Jo P» : Quantitative nutritional studies with water-^ 
solubles chemically defined dietso VIIIo The forced 
feeding of diets each lacking in on© essential amino 
acido Archo Biochemo Biophys* : Sis l|I}.8«l4.55s 1959o 

l\.o Sugimuras To » Birnbaums So Mo j Winits« Mo » and 

^reensteing Jo P* : Quantitative nutritional studies 
with water=soluble» chemically defined diotso IXo 
Further studies on D-glucosatalne^containing dietso 
Arch. Biochemo Biophys» : ^» 521-5279 1959o 

5o Greensteino Jo Fog Oteya '^« Co 9 Bimbaumj So Mo and 
WinitZa Mo : Quantitative nutritional studies with 
water=soluble» chemically defined dietso Xo Formulation 
of a nutritionally complete liquid diet* Jo Nat© 
Cancer Inst© » in pr@sso 

60 Winita* Mo » Birnbaum* So Mo p Sugimura* To » and Oteys Mo C» ' 
Quantitative nutritional and in vivo metabolic studies 
with water-soluble 9 chemically defined dietso 
Jesse Po Greenstein Memorial Volumso Academic Press « 
No Yog in press* I96O0 



U38 



Serial Koo NO 1-339 
PHS-NCI 1^ Lab. of Biochemistry 

Individual Project Report 2, Physo Cheirio and Chroinato. 
Calendar Year 1959 graphy Section 

3q Bethesda, Mdo 



Part A« 



Project Titles The Fractionation of Oligonucleotides 
Derived by Ensymic Degradation of 
Nucleic Acids o 

Principal Investigators: He A. Sober and EoAo Peterson 

Other Investigators; M, Staehelln (Clba fellox^) 

Cooperating Units j None 

Man Years » (Calendar year 1959) 
Totals 2/3 
Professionals 1/3 
Others 1/3 

Project Descriptions 

Objectives s To separate oligonucleotides derived by 
enzymlc digestion of nucleic acids as a means of 
characterizing the nucleic acids or the nucleases 
employed. 

Methods Employed s The digests are fractionated by 
chromatography on celluloslc ion exchangers, using 
gradient elutlon techniques. The resulting fractions 
are examined by physical, chemical, and enzsrmlc methods « 

Major Findings s Oligonucleotides resulting from the 
digestion of yeast and rat liver ribonucleic acids 
by pancreatic rlbonuclease can be fractionated on 
celluloslc anion exchangers, using volatile salts 
(ammoniiam carbonate, acetate, or formate) for elutiono 
The fractionation is based primarily on differences 
in size, but base composition and sequence also determine 
the order of emergenceo Experiments vfith partial 
digests of synthetic polyadenyllc acid indicate that 
decanucleotides (and probably longer chains) can be 
chromatographlcally separated oa DEAS cellulose* 

SEE CONTINUATION SH3ET (A) 

Part B included Yes 



h7^ 



Serial No. NCI«339 

CONTINUATION SHEET (A) 

Significance ; Th® inadequacy of present chemical methods 
for the characterization of nucleic acids and the Inability 
of present assay methods to distinguish subtle differences 
in the specificity of nucleases handicap the study of the 
biochemical processes in which the nucleic acids are 
involved, Th© described method of examining the individual 
products resulting from the partial ensymic hydrolysis of 
a nucleic acid fraction will serve to characterize either 
the ensyme or the nucleic acido Previous chromatographic 
methods J employing ion exchange resins, are limited in that 
components larger than tetra«or pentanucleotides cannot be 
elutedo 



The method will be used in the study of nucleic acids of 
mammalian and viral origlno Consideration will be given to 
the possible use of the data in sequence anaylsis,, A 
homologous series of polynucleotides obtained by this method 
will be used as substrates In the study of certain nucleases, 

The effect of added amino acids and peptides on the chromato- 
graphic behavior of the nucleotides xd.ll be explored in the 
hope that specific tendencies to form x-ieak complexes might 
be revealedo 



4^0 



Serial No„ KG 1-339 



PHS«NIH 

Individual Project Report 

Calendar Year 1959 



Part Bo Honors^ Awards and Publications 

Publications other than abstracts from this project? 

lo Staehelln, Moj Peterson, E, A« , and Sober j, H, A,,j 
Biophyso Blochesio Acta, 8|s 289«292 (1959), 

2o Maver. Mo Eo^ Peterson, E^ A..^ and Greco, A» Eo#i 
"Purification and Characterization of Ribonucleaae 
of Calf Spleen," Ann, No Yo Acado Sci, 81, 599-610 
( 1959)0 This paper was listed as in press in the 
1958 report o 



hU- 



^^ 



Serial Noo HCI»356 
PHS»HIH 1„ Labo of Biecheraistry 

Individual Project Report 2o Phys. Chemo and Chroma* 
Calendar Year 1959 tography Section 



3q B©thesda„ Md< 



Part A, 



Project Title: General techniques and concepts Tor 
the chroraatography of macroinolecules 

Principal Investigator: E, Ao Peterson and H, A, Sober 

Other Investigator: Ro W, Hartleys Jr<, 

Cooperating Units: None 

Man Years (calendar year 1959)t 
Total: 3 
Professional: 1 
Other : 2 

Project Description: 

Objectives ; To devise and prepare adsorbents for the 
chromatography of substances of high molecular weight; 
to develop criteria for testing adsorbents^, such as 
specific adsorption capacity^ efficiency in resolution^ 
etCoi to design and test systems for continuous auto- 
matic recording rtf chromatographic dstaj to devise 
arrangements for the production of various gradients of 
pHj saltg and other variables required for the chromato- 
graphy of macromoleculesj to study the effect of particle 
sizep column geometry^ flow rate^ temperature^ etco on 
the resolution obtained^ and to develop and investigate 
fundaiaental concepts dealing with the processes occurring 
during chromatography. 

Methods employed ; Cation and anion exchangers are made 
from cellulose of different particle sizes by attaching 
ionizing groups having various acidic or basic strength So 
The effects of variables under study are evaluated with 
respect to the resolution of test mixtures of macro-^ 
molecules o Special experiments are devised for the test« 
ing of concepts o 

SEE CONTINUATION SHEET (A) 
Part B included Yes 



442 



Serial Noe MC 1=356 

CONTINUATION SHEET (A) 

Major fi n ding s; Extensively resol-^ed and reproducible 
chromatograms o£ serura or plasraa can b© produced in 6 
hours* Th© limitation on flow rat© appears to b© hydro- 
dynamic in natures chroraatographic resolution vjas not 
affected by the highest flow rates perraitted by the 
laschanical properties of th© adsorbents employedo 

Buffer exchange on Sephadexe a nen comraercial Biat©rial» 
has been evaluatedo It offers many adjutages o'\7©r 
dialysis as a means of equilibrating protein x^ith a 
buffer required at th© start of a chromatographic ex- 
periment© and it brings th© large scale chromatographic 
fractionation of s.erura proteins closer to reality* Work 
in this laboratory has shown that mixtures of Sephadex 
and the eellulosic ion exchangers raak© it possible to 
achieve displacement chromatography with salt as a dis» 
placer* The protein can be applied in high salt concen- 
trations and th© capacity of the coluzsn is far beyond 
that to b© expected from its content of eellulosic ad- 
sorbents as observed in ordinary chromatographyo Used in 
another waye such mixtures of Sephadex and eellulosic 
ion exchangers allow stepwise displacement* which on 
theoretical grounds should be free of some of the serious 
objections inherent in ordinary stepwise elutiono 

i?undamental studies of th© absorption process require 
relatively large stocks of chroraatographically homogeneous 
material but no commercially available protein that has 
been tested has been found to be satisfactoryo Simple 
procedures for the examination of th® suitability of such 
materials have been developed^ 

A commercial instrument for the automatic chromatographic 
analysis of amino acid mixtures has been installed and 
modified to eliminate serious difficulties initially en<= 
countered and is currently being ealibratedo 



SEE CONTINUATION SHEET (B) 



ll lili 



Serial No<, HCI-356 
CONTINUATION SHEET (B) 



SigjalficaiicQ ? The ^^ffectlveneas of chromatography in 
its- various applications to macromolecttles depends upon 
appropriate adjustment to the properties of the sub- 
stances under study and a correct interpretation of the 
results obtainedo Basic studies of the mechanism of 
adsorption and of the factors that determine or permit 
high resolution should provide the relevant information.) 

Proposed course of project t The preparation end testing 
of new adsorbents for us® in the fractinnation of sub- 
stances too tightly adsorbed by present ion exchangers 
will be continuedo Apparatus for automatic stepwise as 
well as gradient elution will be completed. Equipment 
for the continuous. monitoring of effluent streams will 
be evaluetedo The limitations as well as the advantages 
of displacement chromatography on Sephadex-cellulosic 
ion exchanger columns will be further esploredo 

Experiments designed to provide fundamental insight into 
the processes attending the adsorption of macromolecules 
Hill be performed with model compounds comprising homo- 
logous series of increasing size, charge^ snd charge 
distribution 8 as provided by isolated or synthesized 
polynucleotides^ polypeptides^ atce 



[111 in 



Serial Noo HCI-356 

PHS-NIH 
Individual Project Report 
Calendar Year 1959 

Part Bs Henerss Awards, and Publications 

Publications other than abstracts from this project: 

lo Peterson a E, A», and Sober^ H, Aot "Diethylamino^ 

etbjlcelluloa©p" "Carboxymethylcellulose". "BCTEOLA° 
Cellulose "p and "Phosphorylated Cellulose". Bie« 
che mical Preparations 8^ edited by A, Meister, John 
Wiley and Sons^, ^q\j York. In press 1960o 

2o Peterson, Eo A., and Sober^ H, A.: A variable 

gradient device for chroma tographyo A Laboratory 
Manual of Analytical Methods of Protein Chemistry 
edited by Ro Jo Block and Po Alexander, Pergamon 
Press, London^ in press, 1960o 

3o Sober, H, Ao, and Peterson, E^ A^: Chromatographic 

• evaluation of protein mixtures « Greenstein Mem orial 
Symposium edited by J« Edsallo Academic Press, " Maw 
Ifork, in press, I960 

i|.o Peterson, Eo Ao, and Sober, Ho A.j Column chroma- 
tography of proteins: substituted celluloseso 
Methods in Enzymolofgy V, edited by S, Po Golowick 
and No bo Kaplan, Academic Press, New York, in press, 
1960, 

5o Peterson, E. A,^ and Sober, H, Ao: A variable 

gradient device for chroma tographyo Analo Chem* ^t 
857-862, 1959o This paper was listed as' "in press^ 

• in the 1958 reporto 



Iilic< 



Serial Koa NCI»357 
PHS.-NIH lo Labo' of Biochomistiy 

Individual Project Rsport 2o Physi CheiHo and Chroioato- 
Calendar Year 1959 graphy Section 

3«) Betheada, Mdo 

Project Title; Homogeneity and Structure of Proteins 

Principal Investigators? Eo Ao Peterson and Ho Ao Sober 

Other Investigators: R^, Wo Hartley^ Jro 

Cooperating Units: None 

Man Years - (Calendar Year 19^9) 
Total: 1 
Professional: 1/3 
Other: 2/3 

Project Description 

Objectives; To determine the nature and origin of the 
Biolacular heterogeneity observed in proteins having 
what appears to b® the same biochemical activity, and to 
investigate the possibility that many of the differences 
between apparently distinct species of protein might 
be the result of differences in configuration or 
complexing that mask identical or nearly identical 
primary structures » 



Selected proteins are chroma to graphed on cellulosic 
ion exchangers under condit-ions providing a hi^ degree 
of resolution, in the presence or absence of agents 
that break the bonds maintaining secondary aad tertiary, 
and quaternary structure but do not affect the primary 
structureo When possible, chromatography of proteins 
treated with such agents is also carried out after the 
removal of the agents 

Major Findings; The treatment of whole serum with 
tiM urea, followed by the removal of urea by dialysis ^ 
yielded a solution with no evidence of precipitate a 
Gradient elution chromatography under conditions routinely 
used for senun produced an elution pattern somewhat 
similar to that of untreated seruma However, the peaks 

SEE COHTINUATIOH SHEET (A) 

Part B included Yes 



llllQ 



Serial NOo NCI«357 
CONTINUATION SHEET (A) 

were considerably more heterogeneous electrophoretically. 
Chromatography in the presence of i|M and 8M urea has proved 
to be feasible on both DBAE- and CM-cellulose, and fractions 
isolated thereby remained soluble e\'en after urea was removed 
by dialysis o These experiments have served to demonstrate 
that serum proteins can be handled in this manner o 

Simple adsorption procedxires have removed all the dimer frcsn 
commercial crystalline albumin , but careful study has confirmed 
suspicions arising from previous work tliat the monomerlc 
portion, itself, comprises a spectrum of molecular species o 
Kos^s'o^^ej? o when the dimer was concentrated to 70% (frrai an 
initial 5%) 9 a heavier component represented 1S% of the total 
proteino Concentration of the latter to kS% was accompanied 
by the appearance of still anotherj faster sedimentlng 
component o 

Significance ? If the goals of this investigation can be 
achieved^ a better understanding of protein structure and the 
nature of the overall protein-synthesizing apparatus will be 
obtainsdo Moreover, some light may be shed on the processes 
involved in the differentiation of the cell, and perhaps on 
carcinogenesis. If a significant portion of t he molecular 
and functional differentiation observed among proteins is 
found to be based on changes not involving the sequence of 
amino acids in the polypeptide chains « 

A more immediate result may be a clarification, with respect 

to nature 3 origin, and biological significance ^ of the 

heterogeneity already observed in proteins that carry what 
appears to be the same biochemical actlvltye 

Proposed Course of Project; Untreated serum protein has 
been chrom-atographically fractionated, in quantity, into 
several components in order to provide purified material 
for studying the effect of urea treatment on the rQchromato« 
graphy of individual peaks in the presence of and in the 
absence of ureao Since the chromatographic components of 
th© original serum protein undoiabtedly contain more than 
one species J multiple peaks after urea treatment are expected 
in most caseso If peaks obtained by the urea treatment of 
distinctly different native proteins coincide in position, 
they will be examined for evidence of similarity or identity 
in amino acid composition and end groups » All fractions will 
be characterised by electrophoresis and sediment at iono Other 
agents, such as guanidlne hydrochloride, sulfhydryl compounds, 
and veraene will be used in combination and alone<, 



IlIlT 



Serial Noo NGI«357 



PHS-NIH 
Individual Project Report 
Calendar Year 19^9 



Part Bo Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Gutter, F, Je , Peterson, E» As, and Sober, Ho A,,: 
Chromatography of Proteins, III: Hiaaan, Horse, and 
Dog Hemoglobins on Cation-Exchange Cellulose, 
Arch, Biochem„ Biophys. 80, 353«366( 1959 )o This 
paper was listed as "in press" in the 1958 rsporto 

Honors and Awards relating to this project; 

None 



Serial No, NCI«36l 
PHS«NIH lo Labo of Biochemistry 

Individual Ppojsct Report 2o Phys, Chairio and Chrorao-= 



Calendar Year 1959 tegraphy Section 

3.P - Beth©sda« Md. 



Part Ao 



Project Title: Studies on the D=Peptidase Activity 
©f Hog Renal AiaTnopeptidase 

Principal Investigators: Go Letellier ^^^ 

Mp Winitz 
So Birnbatara 

Other Investigators: None 

Cooperating Units: None 

Man Years (calendar year 1959): 
Total: loii- 
Professional: lel^. 
Other : None 

Project Description: 

The optically pure L-L, L-D, D-L^, and D»D stereomera 
of H'Ala-AlaoOHg HeAla»L©u'^01^ l-LeuoATaToH^ HoLeuoLsuoOH, 
H»Leu»Phe«OH^ H.Leu»Val«OH, H^PhesPhe.OHs HeVeloLeuoOH^ 
HoValsPhe-OH^ and HeValeVal'OH were synthesized by 
condensation of the pertinent carbobenzosy«»L (or D)<» 
amino acid benzyl ester via the mixed carbosyliC'>=*^ 
carbonic acid anhydride proeedurej, followed by palla=> 
dium«=cetalyz6d hydrogenolysis ©f the condensation 
product so derivedo 

Studies on the susceptibility of the free dlpeptides 
to the hydrolytic action of hog renal amin ©peptidase 
revealed that the rate of cleavage of the L-L form of 
any given dipeptide was, with the e;sception of certain 
generally resistant peptides of valine^ greater than 
that ©f its corresponding L«.D^ R'-U» ®"^ R^R Stereemero 

(1) Dro Guy Letelller 
38 Rue Bisson 
Hull^ Po Queo» 
Canada 

SEE CONTINUATION SHEET (A) 
Part B included No 



Serial No<, HGI « 36I 

CONTINUATION SH3BT (A) 

However, the rate of cleavage of several of the 
D-D stereomers was not only strikingly rapid, but in 
some instances also appreciably greater than that of 
their corresponding L-D or De.L stereomers. Heat 
denaturatlon studies^indical'e^ that the L- and D« peptidase 
activities reside within the same enzyme'^molecuTee In any 
©vent, the ability of a peptidase to hydrolyze rapidly di- 
peptides which contain two D"ainino acid residues stands in 
marked C(?ntrast to the L«directed storeospecificity so 
generally associated wi¥h peptidase action, 

Significan ceji, Study of the renal aminopeptidas© system can 
thereby increase our knowledge of the mechanism of action 
of biological systems ^ on the one hand, while on the other, 
providing a much needed tool for ascertaining the optical 
purity of the multitut© of synthetic peptides presently 
being prepared© 

Proposed Course of Study ; To study D«.peptidase action 
'in the whole organism in feeding experiments vjith rats 
where one or more essential amino acids are replaced In 
the diet by their corresponding D-isomers which are bound 
in peptide linkage with another amino acid. AlsOj, to use 
aminopeptidase , in vitro, as a means for determining the 
optical purity of synthetic peptides o 



u^n 



Serial Noo NGI°362 
PHS«NIH lo Labo ef Eiechsmistry 

Individual Pi?oject Report 2c Physo Cherao and Chroma- 
Calendar Year 19^9 tography Section 

3o Bethesdfij, Mdo 

Part _A o 

Project Title: Separation of Diastereomeric a-Amino 
Acids by Coluim Chromatography on a 
Macro Seal® 

Principal Investigators: K, Michi ^^^ 

S. Birnbaum 

Mo Winitz 

Other Investigators J None 

Cooperating Units: Nona 

Man Years (calendar year 1959): 
Total: 1 
Professional: 1 

Other: None 

Project Description: 

Similarity of the chemical and physical properties of 
diastereoraers which compose mizttsrss such as isoleu-^ 
cine«allolael0ueine and tbreonine-allethreonine oft- 
times presents considerable obstacles to their separa- 
tion© Although column chromatography has sometimes 
been employed to advantage here, such separations^ 
for the most part^ either have been confined to micro 
amounts of material or tend to give peaks which overlapi 
in addition^ considerable lowering of yield results 
from the association of the products in the eluates 
with large amounts of buffer salts© la the present 
study^ the use of Amberlite CG 120 columns (796 x 
150 cms ) In the ammonitim form and volatile buffers 
(ammoniiam acetate^ formate or bicarbonate in ij.0^ 
alcohol) for ©lution have permitted the complete 
separation of the diastereomeric forms ef several 

(1) Present address: 520^ Saginomiya'=»Jutaku 

Saginomiya 5-305 
Nakano«»KUg Tokyo j, Japan 

SEE CONTINUATION SHEET (A) 
Part B included No 



i|51 



I 



Serial Noo NCI -362 
CONTIOTJATION SH3ET (A) 

diasymmetrio amino acids in quantitlos ranging up to 
3 grama and with recoveries of ptara rmterlal averagj 
between 90 and 95^s. This raethod has proven of particular 
utility for the separation of the optically active forms 
composing the commercially- available epimeric L«.isolQUcine« 
D»alloisoleucine mixture | the degree of opticaT homogeneity 
of th9 latter preparation may also be easily measured 
enzymatically through manoraetric assay by subjecting th© 
separate L«isoleucin© and D«alloisoleucine fractions to the 
action cf 2** ^^^ hr sonino a^ld ozidasej, respectivelyo 

Significance; Use of such systems should greatly facilitate 
separation of the diastereomeric forms of diasymmetric 
amino acids and should make these formerly rare materials 
more readily accessible for biological studiesa 

Proposed course of study ; To examine the utility of this 
system for the macro separation of other diastereomeric 
amino acids, such as hydroxy lysine o 



452 



Serial Noo NCI -365 
PHS-NIH 1^ Labo ©f Biochemistry 

Individual Project Report 2o PhySo Cheaio and Chroma. 
Calendar Year 1959 tography Section 

3o Bsthasdsio Mdo 



Part A< 



Project Title: Investigation of Cytoplasmic Nucleo- 
proteins and Their Role in Protein 
Synthesis a 

Principal Investigators: Eo Ao Peterson and E, Lo Kuff 

also Ro Wo, Hendler of the 
National Heart Institute o 

Other Investigators: None 

Cooperating Units: NHI 

Man Years (Calendar year 1959) 
Total: 1/3 
Professional: I/3 
Other: None 

Project Description: 

Objectives : To determine whether the ribonueleoprotein 
particles ©f rat liver and hepatoma comprise a mixture 
of different species and^ if so^ to investigiate the re- 
lationship of the individual species to the synthesis 
of liver proteins ^ 

Methods employed : Cytoplasmic ribonuclenproteln partly 
cles are isolated by differential centrifugation^ and 
radioactive tracer methods are used to study the chroma- 
tographic behavior of protein transferred from the gross 
particle preparation^ and from fractions thereof obtained 
by chromatography on cellulosic ion ejjchangerso 

Major f indings: Cytoplasmic ribonueleoprotein particles 
of rat liver, labeled in vivo by the injection of radio™ 
active amino acids and'°rsolated by differential ultra= 
centrifugation^ released a portion of the label when 
incubated xiiith unlabeled soluble liver preteinso The 
proteins bearing this label behaved like normal soluble 

SEE CONTINUATION SHEET (A) 

Part B included No 



hs:i 



Serial Roo NCI»365 
CONTIKUATIOH SHEET (A) 



proteins when chroma tographed on DEAE-c©lluloseo On 
the other hand^ labeled protein released by disruption 
of the particles by trsatmsnt with versen© was chroma- 
tographically quite unlike the normal soluble proteins^ 
for it adhered to the column with extreme tanacityo 

Some fractionation of the particles has been achieved 
©n columns of ECTEOLA°c©llulos@ hatring a low nitrogen 
content under conditions that do not cause degradation 
©f the partieleso 

Sign if i cence^; The cytoplasmic ribonucleoprotein particles 
are wicl®l!y' recognized as a vital part of the protein 
synthesizing apparatus of the cell^ and as such they 
merit study o Moreover, there is some reason to believe 
that chemical carcinogens may exert their primary effect 
on faese particles o The information gained and the tech«= 
niques developed in this investigation are espseted t© 
aid in the study ©f both normal and neoplastic growtho 

Proposed course of projects The development of chroma- 
tographic procedures for the fractionation of the ribo- 
nucleoprotein particles and the soluble protein and 
ribonucleic acid of rat liver and hepatoma will be con° 
tinuedo Improvements in the ultracentrifugal portions 
of the procedure will be soughtj, particularly vjith re- 
spect to the treatment of the laicrosomss with agents 
other than deoxychplat© for the purpose of releasing the 
ribonucleoprotein particleso Particles isolated from 
Novikoff hepatoma without the use of deoxycholate or 
or ether surface<=active agents will b© studied chroma* 
tographicallyo Fractions from these and other prepara<° 
tions will be examined to determine whether different 
groups of proteins are synthesized by different typos 
©f particles o 



UHU 



K:\ividiiai Jrrojsct Repoi-i^ ^ir hem., aad v 

Calendar Year 1959 ,,h3 .Section 

— - ■■- - . : -j^ BQth«sda„ Mdo 



Part kc 



Project Title: Structural and Biochemical Investigations 
of Intracellular Components o 

Principal Investigators: So Lo Kuff 

Other Investigators? R„ Po Zeigel (Laboratory of Biology) 

Cooperating Unita: NCI-.Ij.i9 

Man Years - (Calendar Year 1959) 
Total: 1^2/3 
professional: 2/3 
Others 1 

Project Description: 

Ob^^ctive: Investigation of the cytochemical organiza- 
f fon*o5r~nb rmal and malignant cells p with particular 
reference to problems of synthesis or reduplication of 
Intracellular components during turaor growths At present 
the major interests of the project are centered iipon the 
properties of the ribonucleoprotein-containSng components 
of ttJitior cell cytoplasm and upon the spatial and bio« 
chemical relationships between these components and the 
other intracellular components « 

Methods Employed ; The techniques of tissue fractionation 
and of density gradient and analytical ultracantrifMgation 
have been described in previous Annual Reports of this 
projects 

Res ults; (1) Px'ee nucleoprotein particles representing 
nearly half of the tissue ribonucleic acid have been 
isolated by mild methods from homoge nates of Novikoff 
hepatoma 9 The preparations were devoid of membranous 
elements derived from the endoplasmic reticultHsig and 
were composed entirely of spherical particles of several 
size groups. Particles of the most numerous group had 
iniiividual diams^ters of 2lpni.i and a molecular weight of 
abcuc i4.o5 milllono Treatment with deoxycholate j, th© 

SS3 COJiTIlfUATIOK 3ESET (A) 

Part B included Yea 



U5.S 



CONTINUATION SH3ST (A) 

detergent usually Qxnployed in tha isolation of ribonucleo« 
protein particles from Biammalian tissues ,. removed one third 
of the protein from the particles without altering their 
spherical fonno ATP induced dissociation of the Slpmpi 
particles into smaller sub-units at concentrations found- 
in certain rapidly growing tumor tissues o ADP was much 
less effective in causing this dissociationo A paper embody-- 
ing these results is in preparation,, 

(2) The ultracentrifugal behavior of tha free ribonucleo- 
protein particles of the 129(P) ascites hepatoma has been 
studied in relation to the growth rate of the tUMoro There 
was a marked decrease in the proportion of more slowly 
moving particles betv/een the- 2nd day after transplantation 
(mitotic interval 12 hours J and. the 7th day (mitotic Interval 
about 36 hours)*, Analogous changes have bean shown elsewhere 
to occur in growing cultures of yeast as the nutrients of 
the meditiai were exhausted„ The present results seem to show 
that the physical form of the tumor ribonucleoprotein 
particles may likewise be related to the metabolic state 
of the cellSo 

Significance for Cancer Research ; The concept that the 
malignant state may be understood in terras of the structural 
and biochemical organization of the cancer ceHs constitutes 
the justification for cytochemical studies such as those 
daiscribed abovso The pivotal role of rlbonucleoproteins in 
the biO'=' synthesis of proteins is presently accept edo It is 
therefore of considerable interest to relate the properties 
of these components to the metabolic activities of the other 
intracellular elements of tumor tissuaso 

Future Work; (1) In view of the results presented above p the 
relationships between txanoreribonucleoprotein particles and 
the energy-producing enzyme systems of the cells will be 
investigatedo It is planned to study (2) possible physical 
association of glycolytic enzymes with the particlesj and 
(b) the response of the particles to the production of ATP 
by glyealytieandaxi dative phosphorylation inisolated systemSo 
The ascites hepatoma cells ^ vdiich can be iiicubated ^ vitro 
with specific metabolic inhibitors, may provide another means 
of approach to this problemo 

(2) Preliminary experiments have indicated that some ribonu- 
clease activity is associated with the isolated Novikoff 
hepatoina ribonucleoprotein particlaSo The nrture and extent 
of this activity will be studied more fully in an effort to 
evaluate its possibls role in the metabolism of the particles. 



Llc^r 



'^m 



PHS»NIH 

Individual Project Report 

Calendar Year 1959 



Part Bo Honors, Awards, and Publications 

Publications other than abstracts from this projects 

Kuff, E<, Lo, and Dalton, Ao J.,s Biochemical 
studies of isolated Golgi membranes. In 
Subcellular Particles , a symposium held during 
the meeting of the Society of General Physiologists 
edited by To Hayashi^ published by the American 
Physiological Society ^ 1959, PP lll|.-'126o 



I 
I 
i 



Serial NOo NGI=330 
PHS«NIH 1„ Lab„ of Biochemistry 

Individual Project Report 2« Cytochemistry Section 
Calendar Year 1959 3o Bethesda. Mdo 



Jr3.3?X3 Ac 



Project Title: Combined Actions of Anti-Cancer Agents 

and Hoinmonal Systems that inhibit Cancer 
Metabolism and Growthe 

Principal Invsatigatorss Do Burk and Mo Woods 

Other Investigators? Jo Laszlo, Jo Hunter , Ko Wight 

B, Stambuk (Research Fellow) and 
Te Howard o 

Cooperating Units I S^ p„ Vollmerj Endocrinology Sect.^ 

Cancer Chemotherapy National Service 
Center J Mo Landy and Mo Shear ^ Labo 
of Chemical Pharmacology, NCIp 



[ectives; To determine the action and mode of action 
of /inti"Cancer agents when used _in vitro and in vivo^ 
in conjunction with relatively tumor= specific inhibitors 
of glucose utilization, and/or other specific metabolic 
processes o 

Methods ; Tumor materials are either exposed in vitro 
to the compound (s), or eompound^s) is (are) administered 
in vivo^and the effects on in vitro metabolism subsequent" 
ly determinedo Methods of metabolic analysis developed 
in connection with studies of insulin: anti-insulin 
hormonal metabolic regulatory systems are employed o 
A series of tiamors exhibiting a spectrum of honnonal 
sensitivities are employed as test systemso Effects of 
treatments in tumor growth and viability are measured 
with appropriate technics o 

Ma.^or Findings; During 1959 special attention was 
given to the mode of action of a series of fluorinated 
pyrimidinesj and related compounds, on tumor cell 
metabolism and grovjtho The Shrllch and Krebs«2 Ascites 
tumors were used as the principal experimental materials o 

SSB CONTIMJATION SHEET (A) 

Part B included Yes 



i|5S 



Serial Noo NCI-330 

CONTINUATION SHEET (A) 

Comparative studies wsre later carried out with ' normal 
liver and bone marroWg through the courtesy of ii>2>, Robert 
Duschinsky of Hoffmann LaRoche the following compounds were 
made available; 5-fluorouracil (PU)^ 5-fluorocytidine (FOR) 
5- fluor ©uridine (FiJR)^ 5-fluoro-.2»-d0osyuridine (PUDR)^ 
5«fluorocytosin0 {FC)^ and 5»fluoro«2»-deoxycytidine (KJDR). 
A series of normal pyriraidines, pyrimidine nucleosides and 
nucleotides were also tested together with various purines 
and derivatives e Using the Ascites tumors as test systems 
it was found that FUR was by far the most active antiglycolytic 
substance tested^ FOR ranking next, then FUDR and PUo R3 
was inactive as was S^-bromouracilo PGDR produced inhibition 
only after several hours suggesting conversion to an active 
compoundo It is interesting that this spectrum of Metabolic 
activities corresponds rather closely to the reported in vivo 
spectrum of activityo While PCJR, on a weight of compound 
basis, exerts the greatest Anti-glycolytic effect it is also 
the most tosic to the hoste Of the normal analogues ^ uridine 
is the only substance producing pronounced in vitro glycolytic 
inhibition of the Ascites "tumors o 

Ij.OO parts per million of uridine will inhibit aerobic glyco«» 
lysis to about the same extent (50 to 100^) as 20 poPoSi, of 
5-=fluorouridineo The glycolytic inhibition produced by the 
S-'fluorinatsd pyrimldines (or uridine) is strongly counter^ 
acted by raising th© level of inorganic phosphate j> removal 
of oxygen^ or by the uncoupling agents 2,1^. dinitrophenol and 
pentachlorophenolo Glycolytic inhibition which is much 
greater imder aerobic conditions may result from a specific 
interference with rephosphorylation of ADP, thus limiting 
the availability of ATP for glycolysis o Insulin reversibility 
of FU^induced inhibition of glycolysis, which occurs in 
certain tissues sensitive to insulins anti^ insulin hormones 
(Melanoma J bone marrow), does not occur in this Ascites tumors „ 
This effect of insulin would be predicted from the fact that 
the hormone lowei^sthe requirement for ATP in theh©ZQk:inase 
reaction of anti-insulin regulated tissues,, Strains of 
Shrlich Ascites tumors differing in sensitivity to fluorinated 
pyrlmidines were obtained from fi?. Charles Heidelbergero 
These display highly characteristic patterns of glycolytic 
response to a spectrum of pyrimidines^ to adenine and to 

S3S CONTINUATION SH3ET (B) 



IiCT! 



Serial NOo KGI=.330 

COUTIHUaTION SHi5ET (B) 

inorganic phosphatSo Tha datag hM.1© too involved to detail 
her©, ara suggestive with respect to mechanisms of drug 
resistance o Perhaps the most interesting and practical 
findings is tbat the in yitrp anti^metabolic action of the 
fluorinated pyrimidines ( and iiridin® ) depends upon the 
presence of glucose (siiccinate, pyruvate^ glutamate will 
not substitute for glucose) a Furthermore ^ in vivo studies 
in progress indicate that the anti^tumor action of 
5<»fluorouraGil can be markedly increased in certain cases 
by siiiiultaneous injection of high levels of glucose because 
of greater homeostasis of normal tissues j, and relative glucose 
deficiency in many tumors ^ therapeutic applications may be 
possible o Glucose«»dependence of other classes of anti- 
metabolites will be testede 

In collaboratioii with Drs.Landy and Shear of the Labo of 
ChesEical Pharstiacology extensive in vit^ tests of cytotoxic 
serum components have been made with^'various mouse txomorSo 
The cytotoxic complex^ which Involves complement , divalent 
cations and, a factor vJhich may be antibody, has been identi-» 
fied in sera from a number of individuals of a series of 
mammalian and avian species„- Work is in progress aimed at 
delineating "the mechanism of cytotoxic action with special 
refersnc© to specific metabolic systems of the susceptible cells, 
and to further characterization of the active factor(s)o The 
effect of combining specific metabolic inhibitors (or stimu- 
lators) with these naturally occurring serologic systems 
is being studied^ It is planned to bring specific anti«»tumor 
antibodies into the study eventually o 

Proposed Course of Project; Further tests of endotoxic 
polysaccharides (see 19$B "annual report ) are planned in 
collaboration with DrSo Landy and Shear o The combined use 
of the agents with specific antimetabolites and/or serologic 
factors is planned o 



iicn 



Serial NOo NCI-330 



PHS^NIH 

Individual Project Report 

Calendar Year 1959 



Part Bo Honors 3 Awards and Publications 

Publications other than abstracts from this project: 

la LazlOp Jo 9 Burkj D», and Wight ^ Ko : Inhibition 
and enhancement effects of hypochlorite on 
Ascites tumor cell metabolism and growth^ and 
on host resistanceo J, Nato Cancer Insto 23 
35l«36$, 1959 o "^ 

2i Burk, D.g Lasaslo, J,, and Wight, Ko j Comparative 
Metabolism of normal human leukocytes, Pedo 
Proceeds 18 s 199, 1959. 

3o Woods, Mo, Burk, Do and Hunter ^ Jo^: Methotrexate, 
5«fluorouracil, and prednisolone as inhibitors 
of tumor glycolysiSo Proceed o Amere Assoco 
Cancer Research ^: Jh.^ 1959. 

ifo Hunter, J.^ and Woods, MogrSffects of 5«fluoro\iracil 
and prednisolone on the growth of S91 Melanoma o 
Proceado Amero AsaoCo Cancer Research ^j299l959o 

5o Woods, M.J Landy, M,, and Shear^ M^,; Effects of 
endotoxic polysaccharides of tumor glycolysis » 
. Peda rroceed, 18^ 355, 1959o 



HBl 



Serial Noo NCI-331 
PHS-NIH lo Labo of Binchemistry 

ludividtsal Project Report 2o Cytochemistry Section 
Calendar Year 1959 3o Bethesda, Mdo 



Part A< 



Project Title: Subcellular localization of enzymes 

involved in the carbohydrate metabolism 
and growth of tumors o 

Principal Investigators; Be Stambuk (Research Fellow), 

Mo Woods and D, Burk 

Other Investigators: None 

Cooperating Units: Mo Landy and Mo Shearof the Labo 

of Chemical Pharmacology 

Man Years (calendar year 1959): 
Total: 1 / 
Professional: l/j 
Other: 2/3 

Project Description: 

This project was not active during 1959 since neither 
Dro Paul Hochstein (who worked on this project prior 
t© 1958) nor I4ro George Hobby (one of the 'principal 
investigators in 1958) were available o It is planned 
to continue these investigations with Dr© Stambuk in 
I960 with special reference to the effects of complement^ 
dependent cytotoxic seruiji factors on subcellular 
metabolismo 

Significance: An understanding of the precise mechanism 
of drug action against tumor cells requires a knowledge 
of the localization of the affected en2yme(s) within 
th® cello The possible etiologie role of aberrant 
mitochondria, or other hereditai»y particulates of the 
cytoplasm, must be considered in relation to the enzymic 
properties of such structures o 



Part B included: NO 



Serial Noo NCI«332 
PHS=NIH lo Labo of Siochemo 

Individual Project Rspopt 2o Cytochemistry Section 
Calendar Year 19^9 3® Bethesda* Md« 



Project Titl©s Effects of Steroid Hormone Balance 

on Growth of Transplanted Mouse Tumors, 

Principal Investigators s Jo Go Hunter 

Oth©r Investigators s Mo Mo Moods and Do burk 

Cooperating Units: Endocrinology Sections CCNSC 

Man Years (calendar year 1959 )s 
Total; 2/3 
Professional: 2/3 
Other: Non© 

Project description: • 

Obleetiveg; To determine the effects of manipulation 
ofnosFl^riaonal balance on growth of transplanted 
mouse tuniorso 

Met hods: Tumors are available which are sensiti-^e 
to strsss«modifiable hormonal mechanisms (insulin: 
®riiti='insulin system) which affect tumor metabolism 
and groMth via regulation of the hexokinase reactiono 

While th© same hormonal systems strongly regulate 
the metabolism of normal tissues over the same 
mechanisas* glycolytic homeostasis is maintained to 
an extant not possible in tumors© Effects of 
manipulation of host endocrine balance by physical p 
chemical* or biologic means are measured in terms 
of tumor growth rateo 

SEE COUTIITOATION SHEET (A) 

Part B included Yes 



463 



I 



Serial Noo NG 1=332 

CONTINUATION SHEET (A) 

Ma;^or findings: A differential effect of ^-fluorouracll 
pre<lni's"oione "^d a cofobinatlon of the two on the raetabolista 
and growth of the S91 melanoma has been pre-^iouslj reported 
(see NCI"330» 3329 33i{.» 1958). $ Fltiorouracll at th® 
raaxinium tolerated doses had onlj small to raodorats effects 
on growth of th© aelanomao Prednisolone was more effective 
than S^fluorouraeiX but not as effecti-ye as their coasbinationc 
These observations are in line i^ith pr©-?iously obtained 
metabolic data« 

Curreiat studies have been concerned with th® effect of ^-^ 
fluorouracll and ^-fluorodeox^uridin© which Is less toxic 
to th© host^ on th© metabolism of normal tissues » principally 
bon© marrowo Inhibition of aerobic glycoljfsis by 5- 
fluorouracll but not by ^'^fluorodeosyuridine was observed 
with rat bone marrow cellso The differential effects of 
these compounds on th© aerobic glycolysis of bone marrow 
is consistent with their relative toxicity to the host 
when administered in vivoo As in the case of S91 melanoma 
cells g but not EhrTToh ascites cells « Insulin reversed the 
inhibition by ^-fluorouracll of aerobic glycolysis of 
bone marrow (se© also NCI«33i5-)o 

Si^iific eneei These studies help ta furnish a basis fer 
ths^evel opmen t ©f combination therapies in i^hich steroid 
cesnpenisits are involved o 

Proposed course ef project; It is planned t© continue tbo 

pre sent general ' "appr oacH ' usi ng additional steroids and 

oiaber types ©f chem© therapeutic agentso 



461^ 



Serial Noo NC 1-332 



PHS-NIH 
Individual Project Report 
Calendar Year 1959 



Part B; Honors e Awards* and Publications 

Publications other than abstracts from this projects 

lo Hunters Jo and Woods » M« ; Effects of ^^ 
fluorouracil and prednisolone on the 
growth of S91 taelanomao Proco Araero 
Assno for Cancer Research ^i 29 » 1959o 

2o Woods » Moi) Burkd C« and Hunter « Je : 

r^ethotrexate^ 5"'i*luorouracil» and prednisolone 
as inhibitors of tumors glycolysiso Proco 
Amero Assno for Cancer Research ^: 7i^-» 1959° 

3o Woods* Ho and Hunters Jo J Metabolic and growth 

responses of mouse melanomas » Erebs'='2 carcinoraai 
and brain relative to stress e and insulin :anti« 
insulin hormoneso Pigment Cell Biology l^^S^k^^" 

Academic Press Inco # No io « iJo Yo"» i959<» 

(This paper was reported as in press in the 
1958 Annual Report)© 

Honors and Awards relating to this projects 

None 



4B5 



Serial Noo NCI»333 
PHS»NIH 1^ Labo of Biochemistry 

Individual Project Report 2o Cytochemistry Section 
Calendar Year 19 $9 3o Bethesda^ Mdo 



Part A, 



Project Title: Fundamental Oxidative and Glycolytic 

Metabolism in tiamors and Tissue Cultures 

Principal Investigators: Do Burk and Ko Wight 

Other Investigators: J« Laszlo (Research Fellow )» Mo 

WoodSs Bo Stambuk, To Howard^ Jo 
Hunter, and Go Hobby 

Cooperating Units: Mo Lane ^ General Medicine Branchj, NCI 

Man. Years ( calendar year 1959 ) t 
Total: 1-1/3 
Professional: 1 
Other: 1/3 

Project Description: 

Objectives ; The purpose of these studies was to define 
the effect of antimetabolites and other chemotherapeutic 
agents on the metabolism of s®l^ obtained from patients 
with the various forms of leukemia and other cell typeso 

Methods : Cellular preparations were obtained from 
patients who had a great predominance of blast cells 
arid abnormal lymphocytes in their peripheral blood o 
These were separated from red cells by means of the 
fibrinogen sedimentation technico Glycolysis and '^xygen 
uptake of cells were measured in Warburg manometers in 
the presence of serum^ glucose^ bicarbonate^ and the 
drug under analysiso Various animal and algal cell types 
were also employed o 

Major findings : Moat workers have reported that normal 
human leukocytes have an "unusual" metabolism amongst 
normal tissues^ namely^ a high rate of aerobic glycolysis 
and a high ratio of aerobic glycolysis to respiration^, 
higher J, indeed^ than in the case of leukemic leukocyteso 
However^ nearly all such work has involved use of either 
leukocyte homogenates instead of leukocyte cells, salt 
solutions instead of sera as suspending media, or single 
instead of various (comparative) methods of leukocyte 

SEE CONTINUATION SHEET (A) 
Part B included Yes 

4RR 



Serial Woo NCI-333 

CONTINUATION SHEET (A) 

anticoagulation and isolationo We have studied intact 
leukocytes from some 20 normal humans, using isologous 
(or ad-mised homologous) sera as suspending mediaj 
various msnometric procedures (2»vessel raanometryj 
Summerson differential manometry^ etco)j he^iarin or 
citrate or def ibrinogenation with beads for anticoagula- 
tion! centrifugation (200 g for 1-3 minutes) or sedi- 
mentation (fibrinogen) or simple standing as isolation 
procedures o With 2«»v®ssel manometry^ nearly all Q^^ 
Telues ranged from « 5 cmm acid produced aerobicslly 
/7o$ million calls/hro V/e conclude that normal human 
leukocytes under ln_ vivo«airaulated conditions ordinarily 
have a small or negligible aerobic glycolysis o Qf 
values ranged from 10 - ZS citim acid produced anaerobically/ 
7o5 million eells/hro Glycolysis was stimulated at times 
up to 100^ by insulin end/or polysaccharide endotoxin ^ 
without respiratory inhibltionj, as also found with mouse 
melanoma S91& 

Hypochlorite (500 poPomo ) Inhibited the respiration and 
glycolysis of Kreb3'a2 and L1210 mouse ascites cells more 
than 90 per cent^ and rendered the cells totally non<» 
viable in Inoculation teats o Hypochlorite gave much 
less inhibition of Kr0bs«2 carcinoma slice metabolism 
and even increased the respiration of normal liver sliceso 
A single intraperitoneal injection of inorganic sodium 
hypochlorite solution (Clorox)^ or of organieally bound 
hypochlorite (Clorpactin XCB)^, gave greater than 100 
percent increases in median survival time (MST) of mice 
previously inoculated with Krebs°2 ascites cellso The 
effect was optimal when approximately 2 mgo hypochlorite 
in 0o5 cco water was administered 15 minutes post-ino- 
culation ^ and diminished to about 2S per cent increase 
in MST when given Zh^ hours port-inoculationo Supra^ 
optimal doses of hypochlorite enhanced tumor growth rat© 
and dissemination^ as did repeated daily administration o 
Host prstreatm.ent with hypoehloriteg, given intraperi- 
toneally ©r subcutaneously, similarly enhanced tumor cell 
growth aiKi virulenceo Tumor growth enhancement was not 
specific for hypochlorite but \?as also observed upon pre- 
treatraent with either fermalin^ methotrexatep total <=body 
irradiation, or host temperature stress for 2ij. hours at 
37°Co Hydrocortisone or prednisolone pretreatanents did 
not enhance but retarded tumor growth » The effects ob- 
served with Krebs-2 ascites wore also found with the Potter 
lymphocytic leukemia 3889 which arose in DBA mice and was 

SEE CONTINUATION SHEET (B) 



Serial Noo NCI-333 
CONTINUATION SHEET (B) 

carried in ascites form in BALB/c x DBA/2 (Pj^) hybrids o 
The mechanism of ttmior groi-Jth enhancement produced by a 
variety of chemical and physiological agents thus ap- 
pears to involve more than loss of imm:une response o 
GroMth enhancement with hypochlorite might not occtar in 
topical tr®atiaent of ©ancer cells^ as in surgical woundSj, 
in sufficiently large hosts (humans),, 

Leukemia cells susceptible and resistant to 8-azaguanin© 
have bsen studied with regard to direct metabolic changes 
in respiration and glycolysis produced by this drugo 
8'=aEaguanina caused a prompt and marked increase in gly- 
colysis in vitro of susceptible cells but not of resis- 
tant celTi'^"witH" concentrations of drug that ranged from 
about on©'=»f0urth to four times the pharmacologic level 
eoramonly employed ($0 mg/kg)o Respiratory inhibition 
was observed in both types of cells j, but at lower doses 
in the susceptible cells than in the resistant lineo 
The metabolic effects observed in^ vitro tended to disap« 
pear after repeated daily exposure of the cells to th® 
drug in vivoa, and did so within a transplant generation o 

The action of Cytoxan on the metabolism of susceptible 
mouse leukemia L-1210 cellsj, K2 ascites cells, and 
Ehrlich ascites cells treated in vivo and then removed for 
in vitro analysis^ or treated only ^ vitro, has shown 
pronounced inhibitions of, glycolysis, aeroSic ally and 
anaerobically and also of respirationo The inhibitions 
were very striking after i|. to 72 hours in vivo treatment 
with a single dosage of 200-300 mg/kg cyToxan, the gly- 
colytic inhibitions often approaching 100 percent (on 
a unit cell volume basis )» With a Cytoxan resistant 
strain of L-1210 leukemia respiration and glycolysis 
was, on the contrary, not inhibited but at times stimulatedo 

A strain of theiroophilic algae (Sorokin and Myers 7-ll=» 
05s, optimum temperature 39®C) caused marked inhibition 
d^-O^lOO^) of anaerobic and (especially) aerobic glycoly« 
sis of Ehrlich and K2 mouse ascites cells when mixed in 
approximately equal volumes of cells in Krebs-Ringer 
mediumr. The algae consume glucose at a rate even several 
times faster than the cancer cells (on a unit cell volxjme 
basis), but the glycolytic inhibition is observable long 
before ti^ glucose has disappeared^ The cause of the 
Inhibition is under investigation o The algae, like the 
tumor cells, can produce lactic acid from glucose, but 

SEE CONTINUATION SHEET (C) 



468 



Serial Noo NCI-333 

CONTINUATION SHEET (P) 

D-lactic acid instead of L-lactic acid is formed o A 
single intraperitoneal dose of washed algal cells is 
lethal to normal mice at about 1000 mg wet weight 
{250 mg dry weight) per kg (ioO,^ somewhat less than 
with Cytoxan )p but the tolerance level Is somewhat 
increased by the presence of cancer cells in the intra- 
peritoneal tracto 2ij.«hour pre-inoculatinin of mice with 
sublethal doses of alga© produced a certain prolonga= 
tion of life of mice then inoculated with tumor cells o 
This is the opposite of the growth enhancement effect 
obtained by retrsatment of t\araor»bearing mice with Clorox 
as discussed above. The algae cells have been supplied 
to other workers in the NIH and other Washington labora- 
tories for purposes of preparing unlformly'-labelled C-*-^ 
of high radioactivity^ and in one instance for preparing 
hi^ly radioactive beta-acarotene for use in studying 
vitamin-A formation in the National Heart Institutes 
About fifty cultures a year are supplied (upon request) 
to various university and industrial laboratories and 
investigators o These (thermophilic) algae are the only 
known species of green plants potentially capable of 
being engineered t© purify air (remove COo and produce 
Op) 9 to provide a large amount of required food and to 
eliminate waste products (urine and feces) of numbers 
of humans maintained in confined spaces such as submarines 
and space ships ^ or underground compartments o 

SigDificaaoe ; These studies are aimed at ascertaining 
and unders^nding metabolic peculiarities ef malignant 
cells, under conditions approaching in variable degree 
the in vivo state, and under conditions of chemotherapeutic 
action, "wTSi added attempts to discover new agents of 
the latter type by lof>king first, not lastj, for metabolic 
indication of potential chemotherapeutic action, as X'/ell 
illustrated by the striking metabolic e,'ffects on txanor 
metabolism obtained with algal cells, the mechanism of 
which effects yet remains to be worked outp 

Proposed cours e ©f action t These studies have been and 
will be carried on"by classical research methods, by 
proceeding from lead to lead, from one finding to the 
next, on preferably a day-to-day basis, rather than 
on a Ibaois of relatively non-selective, routine results 
planned for in advance <> 



46S 



Serial Koo NCI-333 



PHS-NIH 
Individual Project Report 
Calendar Year 19^ 



P art _ Bq Honor a^ Awards j, and Publications 

Publications other than abstracts from this project: 

lo Lasslo^ Jog Burk, Do, and Wight, KoS Inhibition and 
enhancement effects of hypochlorite on ascites tumor 
cell metabolism and growth, and on host resistanceo 
Jo Natlo Cancer Insto 2^.: 351-365, 1959o 

2q Laszlo, J., Wight, K^, and Burk, DoS Inhibition and 
enhancement effects of hypochlorite on tumor cell 
growth and host resistanc©© Proce Amero AssoCo 
Cancer Research, ^: 35, 1959o 

3o Burk, Do, Laszlo, J©, and Wight, Ko: Comparative 
metabolism of normal htnnan leukoeyteso Pedc ProtJo 
18: 199, 1959 o 

l|.o Burk^ D,, LasElo, Je, Hunter, Jo, Wight, Ko, and 

Woods, MoS Differential metabelie responses of sus- 
ceptible and resistant mouse leukemia cells to 
8==azaguanln©o Jo Nato Cancer Inste, Jan» 1960o 

5o B'urk, Dc J,- and Hobby, G« : Closed cycle air purifi- 
cation with algaeo Go ProCo lat International 
Symposium on Submarine and Space Medicine, Pebo 1960^ 



470 



PH5:-NIH Serial Noo NCI=.33ij. 

Individual Project Report lo Labo ©f Biochemistry 
Calendar Year 1959 2o Cytochemistry Section 

3o Bethesdas Mdo 



Part A, 



Project Titles The occurrence and function of a stress= 
modifiable anti^lnsulin hormonal complex 
regulating glucose utilization in cancer 
metabolism and growtho 

Principal Investigators: Mo Woods and Do Burk 

Other Investijgatorss Bo Starabuk (research fellow )j 

Jo Hunter and T, Howard 

Cooperating Units: Eo Po Vollmerg Endocrinology Sectieng 

Cancer Chemotherapy National Service 
Center? Mo Landy and Mo Shear^ Labo 
of Chemical Pharmacology ^ ^CI 

Man Years (calendar year 1959); 
Total: 1 1/3 
Professional: 1 
Other: 1/3 

Project Descriptions 

Object lyes; To develop in vitro and in vivo procedures 
F^^definlng qualitatively and quant iTatTvery the opera^ 
tion of the insulinsanti^insulin hormonal mechanism 
which regulates glucose utilization^ and to apply these 
procedures in developing means of diagnosis and selective 
necrotization of tumors^ especially in conjunction with 
multiple procedures for therapyo 

Methods employed; Mice bearing tumors of known degrees 
of sensitivity' to anti«lnsullns Insulin honaonal regula- 
tion are exposed to normal or stressful temperatures and 
the txaraors from such animals used for assay of the effects 
of ©xogeneus . glycolytic, and/or^ respiratory inhibitorsg 
"uncoupling agents" or other substances of potential 
chemotherapeutic intereato 

SEE CONTINUATION SHEET (A) 



Part B included Yes 



hll 



I 



Serial No, NCI<=33lj. 
CONTINUATION SHEET (A) 

Major findings; During 1959 major emphasis has been 
placed on a continued study of the mechanisms by which 
the 5-fluorinated pyrimidiaes act to inhibit glyco- 
lysis in insulin-sensitive and insensitive tumors 
(see reports for projects NGI-330, 332 and 331) » It 
appears that the primary metabolic action of 5-i'luor- 
uracil Cor derived compounds) involves an inhibition 
of aerobic phosphorylation^ thus lowering th® steady 
state level of ATP and so retarding glyeolysisp In 
a tumor like S91 melanoma^ where a considerable anti" 
insulin control of hex'ckinase esists^ any lovjering~ of 
the steady state level of ATP would accentuate this 
restraint of hexdkinase activity^ ThuSg insulin^ 
which lowers the requirement for ATP could be expected 
to counter actg to a variable ©stent, the fluorinated 
pyrimidinee.induced glycolytic inhibition o In keeping 
with thiSg insulin was found to counteract S-flworouracil^ 
induced glycolytic inhibition in S91 melanoma^ and normal 
bone marrowp but not in the insulins anti-insulin insensi«* 
tivs K==2 and Ehrlieh Ascites tumorso 

Work began several years ago on metabolism of tissue 
culture clonesj, in collaboration with D^oKatherine 
Sanferd and Wilton Earle^, has been brought to publica=> 
tioHe The findings show that development ef a high de- 
gre© of raalignaney (high percentage ef takes on trans- 
plantation^ short induction period^ and enhanced capa<=> 
city to grow in non^irradiated mice) is associated with 
a greatly increased glycolytic capacity and lowered 
sensitivity to glycolytic inhibition ef the "anti-in- 
sulin" types 

The precise basis for the anti-cancer action of steroids 
has not been establishede While inhibition of steroid- 
sensitive transhydrogenases, linking DPN with TPNe 
dependent isocitric dehydrogenase ^ may be involved in 
some instances^, there is considerable evidence that 
the hexokinas© locus constitutes a major steroid-sensi- 
tive locus of metabolic and growth controlo Differential 
rates of intra tumoral metabolism may determine which 
steroids are able to accumulate in amounts sufficient 
to inhibit glycolysiso Thus^ testosterone under in 
vitro conditions is more inhibitory to mouse melanoma 

SEE CONTINUATION SHEET (B) 
Part B included Yes 



472 



Serial Noo NCI«33l|. 

CONTINUATION SHEET (B) 

glycolysis than prednisolone, but when given in vivo 
only pradni solone is highly active o ' 

Recently considerable interest has developed concern- 
ing ^« testolelactone^ since this steroid, which is 
virtxnally withoiat endocrinologlc activity, has shoxra 
laarksd antitijinor action in human mammary carclnomao 
Experiments begun late in 1959 have shoxiin that ^ ' 
tsstololactone inhibits in yi tro an aerob i c glycolysis 
in S91 melanoma, but not^Tn the antl«insulin-insensitive 
Ehrlieh Ascites tumoro The inhibiting action, hovjever, 
is considerably belou that of testosterone., Under 
in yi tro aerobic conditions A' testololactone caused 
marked lowering of the Pasteur effect, probably as a 
result of damage to tumor respiration c. In this respect 
the action of ^' tsstololactone resembles that of 
testosterone or diethylstilbes trolo Intraperitoneal 
administration of ^' testololactone gave an average 
loii^ering of in vitro aerobic acid excess of 2)4.% (10 
tiaraor bearing anlraiTs), but did not significantly 
change the anaerobic glycolysis,. Further I'Jork is 
needed, but the results obtained thus far establish 
that this steroid derivative, which is vjithout known 
endocrinologlc activity, nevertheless produces sign!- 
f leant metabolic changes that might be a basis for anti° 
tumor action o 

Si^lf Icance ; Information obtained helps to clarify 
^tEelSasis for the snti-tumor action of steroids and 
their role in multichemotherapeutic systems, especially 
those involving the 5°i'luorinated pyrlmidinesp 

jroject: It is planned t© continue 



rroppseq,^ course^^^ ox_^ pro j e cii; iii as pxannea to coaiij 
ai6ng"l;h'e same general lines, but introducing nev? 
compounds and tumors as indicatedo Particular attention 
X'^ili be given to the metabolie basis for sequential 
therapy with two or more agents o 



i|73 



Serial Noo NCI 331^. 



PHS^NIH 
ladividual Project Report 
Calendar Year 1959 



Pert B; IfonorSg Awards, and Publications 

Publications other than abstracts from this project: 

lo WoodSj M9 and Hunter^ Jo : Metabolic and growth 
responses of mouse melanomas, Krebs«2 carclnomaj 
and brain relative to atress, and insulin: anti- 
insulin hormones » Pigment Cell Biology hS5'°kBQo 
Academic Press Inco^' i^.V!",' N. Yo^ 19^9 1 (This 
paper was reported as in press in the 1958 
Annual Report )o 

2c, Moods, Meg Sanford, Ko^ Burk, Do and Eerle^ ¥<>: 
Glycolytic properties of high and low sarcoma'^' 
producing lines and clones of mouse tissue 
culture cellSo Journ. Nato Cancer Institute 
In press, 1959. Vol. 2^, 1079=1088^ (1909) 

3e MoodSj Mo J Burkj Do, and Hunter, Jo: Methotrexate, 
5«=fluprouracil£, and prednisolone as inhibitors 
of tumor glycolysis e Proceed, Amero Assoco for 
Cancer Reso X' "^^9 1959* 

l{.o Woods, -Mo and Hunter^ JoS Effects of 5-fluorouracil 
and prednisolone on the growth of S91 melanomao 
Proceed© Amero Assocs for Cancer ReSo 2* ^9, 1959o 



^Ih 



Serial Noo NCI-320 
T ^^ -^ PHS-NIH lo Labo of Biochemistry 

Individual Projoct Report 2o Enzymes end Metabolism 



Calendar Year 1959 Section 

3o Bethesda, Md, 



Part A, 



Project Title: Studies on the rates and kinetics 

of enzyme synthesis and destruction 
in vivQ o 

Principal Investigatora: Vincent Eo Prie© and 

Miloslav Rechcigl, Jro 

Other Investigators; None 

Cooperating Units: None 

Man years (Calendar year 1959): 
Total: 1-1/2 
Professional: 1/2 
Other: 1 

Project Description: 

Objective : "^o develop simple techniques for measuring 
the rates of enzyme synthesis and destruction in^ vivo ; 
the kinetics of these proceases| and how they change 
during development^ and how they are altered in disease 
stateso 

Methods employed : The basic technique consists in 
injecting 3'=amino'»l,2j,Ij.°triazole in a dose of 1 gram 
per kilogram intraperitoneally into groups of animals^ 
At various periods thereafter the animals are killed^ 
the liver homogenized in ij.9 volumes of cold watej^ and 
catalase assays are performed on the homogenates 
using a Gary Recording Spectrophotometer with a 
Log-absorban@@ attachment o 

Major findings : In previous reports it was sho>m 
ihat the rapicl fall in catalase activity following 
the injection of 3•=amino<-l,2,i^.°triazole resulted 
from dest?»uction of the cataksej, and that the return 
in catal'ase activity during the following days resulted 
from the synthesis of new catalaseo It was further 

SEE CONTINUATION SHEET{A) 
Part B included: NO 



Serial Noo NCI-320 
CONTINUATION SHEET (A) 

shown by use of radioiron that administration of the 
triazol© produced no significant elteration in the rate 
of catalase synthesis o 

Kinetic studies have been carried out on the rate of 
catalase return following adinlni strati on of amino triazole 
to ratSo It has been shown that the curve can be 
described by the equation: 




where C is the catalase activity at any tirae,t; ^ is 
rate of catalase synthesis per unit time, end k, is 
the fraction ©f catalase destroyed per unit tlmeo 

When this equation is calculated fer 5^520 ^ Ca X C) 
male rats vieighing approximately 230 gmo a k of ko^ 
and k^ of o021ij. are obtainedo This meens that k.o% 

units or 2606 micrograms of catalase x^^ere being synthesized 
each hour per gram of liver and Zolk.% ©f the catalase 
molecules present at a given time are being destroyed 
during the next houro With Sprague-Dax^ley male rats 
weighing appr<^ximately 210 grams the rate of catalase 
synthesis was 2ls.o5 micrograms per hoxar in the liver and 
606 micrograms per hour in the kidney^ and 2o27^ of the 
catalase present was being destroyed each hour in the 
liver and 2ol3^ in the kidney o This means that the rate 
of synthesis per gram of tissue was I4. times as high 
in the liver as in the kidney but the fraeticgj of catalase 
destroyed per unit of time is nearly the same in both 
tissueso The kinetics indicate that a nswly«formed 
catalase molecule does not have a finite life span, 
but instead has the same risk of destruction in each 
unit of time as an older molecule o This is not unexpected 
since most data on the incorporation of Ci[j.-araino acids 
into proteins provide a similar interpretationo 

SEE CONTINUATION SHEET (B) 



476 



II 



Serial Noo NCI=320 
CONTINUATICN SHEET (B) 



It should be emphasized that the amino triazole technique 
has both advantages and disadvantageso Until now the 
principle technique by vrhich the rate of protein synthesis 
and destruction could be estimated was by use of radio- 
isotopes., This necessitated rather extensive fractionation 
at each experimental point in purifying the given protein 
to a constant specific activityo With the aminotriazole 
technique this has- not been necessary, for one has only 
to prepare a homogenate for catalase assayo 

The aminotriazole technique is complicated by the fact 

that one is injecting a drug which does alter the body 

metabolisnio Although radlolron studies indicate that 

the single injection of aminotriazole has little or no 

effect on the rate of catalase synthesis, the kinetic 

data do sl»w that for a period ©f 7°12 hours ^ depending 

on the organ end strain of animal^ any catalase that 

is formed will be destroyedo Similar periods would 

presumably exist for tryptophan peroa-idase, and with 

the "iodine peroxidase" of Alexander in the thyroid 

this period may be somewhat longer because of the apparent 

greater sensitivity of this ©azysie to aminotriazole o 

Although th& net effect of these changes on the rate 

of catalase synthesis in the liver appears to be negligibls^, 

it will be wise to check highly significant points by 

the use of radlolron and radiocarbon technlqueso 

An even greater problem in the aminotriazole technique 
is posed by the finding that in tumo3?=-b earing animals 
there Is apparently a prolongation in the time of amino- 
triazole excretiono As yet this is not based on actual 
determinations of aminotriazole, but is inferred from 
radlolron studies in which more radlolron was incertsorated 
into catalase than would have been predicted from t';® 
low level of catalase activity observed in catalase 
isolated from these anlmalso This area requires further 
investigation. It appears that frogs may have a slrailar 
problem in aminotriazole excretion for after a single 
injection the catalase activity returned very little, 
if at all after 9 dayso This was foiand in experiments in 
which we were attempting to compare the rates of catalase 
synthesis and destruction in different orders of vertebrates. 

SEE CONTINUATION SHEET (C) 



kl- 



Serial Noo NC 1-320 
CONTINUATION SHEET (C) 

The above disadvantage can b© overcome in many instances 
by introducing a given variable after the amlnotriasole 
has been ©xcretedo An example of this, is shown in 
studies designed to determine the rate of catalase 
synthesis and destruction in the starving aniraalo In 
this case normal rats were divided into l\. groups and 2 
of the groups were injected with amino triasole and 2 
were kept as controlso At the end of 18 hours, catalase 
assay showed that the expected rat® of synthesis was 
occurring, and at this time food was reyaoved from one 
of the control and one of the eminotrlazole groups o We 
were surprised to find that the rate of catalase synthesis 
per gram of liver remained at normal S-evels over the first 
lj.=»5 days of starvation^ but the liver lost $0% in weight 
and ij.0% in nitrogen o Thus there was no specific loss 
in catalase synthesis as the result of starvation, but 
instead a general reduction in liver sizeo Since the 
number of cells in starvation is said to remain constant 
and there is known to be a marked loss of liver RNA^, one 
wonders whether the liver cell rapidly responds to starvation 
by loss of synthesizing units^ but thst those synthesizing 
units which remain operate at full activity o This appears 
to be a promising area for future invest igationo 

In the control animals which were starved there was actually 

some increase in the liver catalase activity per gram of 
tissue during the period from 21^'-ij.8 hours^ suggesting that 
as the liver coll shrinks in size the catalase concentration 
within the cell rises until it can be destroyedo Wit^ a 
higher concentration ^ the amoxant of catalase destroyed per 
unit tissue could increase although the fraction of catalase 
molecules destroyed during that time remained constant, 
as would be predicted from the amino triasole curves o 

After S^6 days the starving animal enters a second phase 
in which the rate of catalase synthesis per gram of liver 
starts to fall and this continues progressively until the 
death of %he animal on the Tth to 10th dayo 

Significance to Biomedical Research and the Program of 
the National Cancer Institute ; Although considerable 
attention is being given to the sj^uthasis of proteins 
relatively little is known about their destruction o In 
problems on cachexia in the tumor-bearing host it is 

SEE CONTINUATION SHEET (D) 



478 



Serial Noo NCI"320 
CONTINUATION SHEET (D) 

Important to develop a fund of knowledge about the 
destmaction ©f proteins so as to understand the more 
intimate nature of the cachectic processo Although 
it would be desirable to start with proteins such as 
myosin which contribute to a large fraction of the 
body p2'®toin^ the availability of the aminotriazols 
technique made catalase and the related peroxidases a 
more attractive starting polnto Since the reduction 
in catalase activity of the liver and kidney is one of 
the most prominent features of the tuaior<=>besring host^ 
studies on the kinetics of synthesis and destruction of 
this enzyme are of particular importance in the cancer 
f ieldo 

Proposed Course of Research ; It is of particular interest 
to compare the rates of catalase synthesis and destruction 
in animals of different age, sez^ strain <> and species 
to determine their constancy or va-Mability in these 
parameterso The extent of their aS.toration after injection 
©f cortisone or hydrocortisone which are known to markedly 
lower the catalase activity within the liver ^ will also 
be examinedo 

In relation to the work on starvation it will be important 
to extend these studies to animals on prot©in<=fr©e diets 
to determine the extent to which the results observed in 
the starving animal are due to amino acid deficiency and 
to what extent they result from lack of the energy required 
for synthetic processeso 

Certain substances such as Sedormid are known to inhibit 
catalase synthesiSo Other substances such es ethionine 
and toxohormone fractions from tumors^ some of which have 
recently been purified to a high degree ^ may also inhibit 
catalase synthesis o The effect of these three aubstanees 
will be compared o 

It is known that 3°amino-l^ 2gl{.<= triazole is active against 
tryptophan peroxidase and a peroxidase in the thyroid that 
is involved in the iodination of tyrosine to produce 
thyroxine o Auerbach end cowy^kers have reported that 

SEE CONTINUATION SHEET (F.)' 



kl3 



Serial Noo iJC 1-320 

CONTINUATION SHEET (E) 

aminotriazel© apparently inhibits the synthesis ©f 
tryptophan peroxidase^ ^""t examination of their data 
shows that inacti-^ation of th© enzyme and partial 
resynthesis of new would also explain their datao 
It would be of considerable value to examine both of 
these enzymes to see if the action of aminotriazole 
is similar to that on catalas©, and, if so, to further 
extend some of our studies to these enzymes alsoo 

Although the present studies have been limited to a 
certain family of enzymes it is possible that other drugs 
may bind rather specifically to other enzymes So as 
to rather abruptly eliminate their activltyo If so 
it is possible that similar studies could be extended 
to these enzymeso In this regard it will be of particular 
interest to examine some of the many compounda known to 
inhibit nucleic acid synthesis and protein synthesis «, 



480 



Serial Noo NC 1-3142 
PHS«NIH lo Lab» of Biochemistry 

Individual Project Repox»t 2o ^nzyraes and Metabolism 

Calendar lear 1959 Section • 

3o Bethesda* Mdo 



Part Ao 



Project Title; Studies on Iron itetabolism in the 
Turaor-b earing Hosto 

Principal In-^estigators: Robert E» Greenfield 

Other Investigators: Vincent Ea Price 

Mary Washko 
Betty Whitaker 

Man Years (calendar yesu? 1959 )? 
Totals i-1/2 
Professional: 1/2 
Other: 1 

Project Description: 

Objectivea: Previous studies (se© NCI 3li.l«1958) 
of this "series have shown that the anemia associated 
with any one of a number of animals tumor could be 
explained primarily by a loss of red cells into 
the tumor and indicated tha,t in various tumors 
different vascular defects accounted for the loss of 
erythrocyteso Those tumors associated with the aost 
rapid onset of anemia* such as the rat lymphosarcoma 
2788 » were foisnd to have a narrow band of hemorrhage 
between th© tumor and the connective tissue of the 
capsules In the connective tissue adjacent to the 
hemorrhage* many macrophages were foynd containing 
high concentrations of iron in the form of hemosideriao 
Other tumors that were associated with a more gradual 
onset of smemia showed evidence of hemorrhage into 
th© necrotic areas » as seen in rat hepatoma 3683 # or 
the trapping of blood in thrombosed sinusoids as seen 
in the spontaneous mammary tumor of C^H miceo 

SEE CONTINUATION SHEET (A) 
Part B Included: Yes 



481 



Serial Noa NCI«3lp. 
CONTIMJATION SHEET (A) 

la each case* irrespective of the histological findings* 
a major fraction of the iron lost from the red cells 
diaring the development of the aneraia could be recovered 

in the tumor-o 

As a result of the above studies » a question has arisen 
as to how long iron from ©xtravasated blood remains in 
the area of the tiamoro Is it sequestered in the tumor 
for a long period of times or is it quite rapidly returned 
to th© active iron pool of the bodyt Since the loss o^* 
red cells into the area of the tumor is a progressive 
process which increases with the growth of the neoplasms 
it is difficult to obtain any precise estimate of the rate 
at which the erythrocyte iron which is deposited in th© 
tumor at any given time again becomes available for 
erythrosyt© syntaeslso Because of the complexity of the 
problem in tumors « a simpler system was required in 
which the number of variables could be more readily 
controlled^ Therefore studies were first carried out 
to determine the fate of iron from intact and lyzed 
erythrocytes when injected into the gastrocnemi\js musi'-le 
of normal rats? Subsequently^ th® fate of iron from blood 
injected subcutaneously and directly into tumor tissue 
was investigated© 

Methods employed? In initial experiments* intact and lysed 
erythrocytes labeled with radioiron were injected into 
the gastrocnemius muscle of normal animalso At appropriate 
intervals* groups of two or three animals were anaesthetized 
and a sample of blood was obtained from each on which the 
radioiron concentration was determined. These animals were 
killed immediately after the sample was taken* and the 
livers and spleens were removedo The hind legs were 
carefully dissected from the body* and the remaining carcasses 
were minced© The tissues were subsequently digested by 
heating in 30^ KOH* and aliquots of the digests were 
weighed into vials on which th© concentration of Fe59 
was measured with a scintillation coiuatero In the later 
experiments* the distribution of radioiron obtained 
following the injection of labeled cells directly into 
small tumors was compared with that obtained following 
the subcutaneous injection of labeled cells into normal ratso 

SEE CONTimJATION SHEET (B) 



48 



o 



Serial Noo NC 1-31^1 
CONTINUATION SHEET (B) 

^^P^.X^.^l^B^* c-Q^®" intact erythrocytes which had been 
labeled witfiTFe-'" were injected into the gastrocnemius 
musqls of normal rats» the radioiron left the site of 
injection vary sloMlye at a rat® of 2^ per dayo After 60 
days only iQfo had been reutilized for the synthesis of 
new red cellso Th© i*ate at which radioiron from labeled 
cells left th® injection site in the leg was only slightly 
increased in tiamor^bearing animals or in animals which had 
been mad© anemic by repeated hemorrhageo In contrasts when 
erythrocytes containing F059 labeled hemoglobin were lysed 
and injected into the gastrocnemius muscle of normal ratse 
55 percent of th® radioiron had left the injection site 
within 2i|. hours and 35 percent was recovered in the llvero 
In later experiments » a slow rate was found when intact 
cells labeled with Fo59 were injected into the tvsaior tiasu© 
of tumor^bearing animals and into the subcutaneous tissue 
of normal aaimalso Th© slow reutilization of iron from 
injected blood in these experiments suggests that the iron 
from ©xtra^asated blood may also be only slowly reutilized 
in normal and tumor^bearing ratso 

S dgnificanc© ; Pre^^ious studies have shown that in tumor- 
bearing animals loss of cells through vascular lesions in 
the area of the tumor is a major factor in the pathogenesis 
of the anemiao It is of importance to our understanding of 
tumor»host relationships to learn how rapidly hemoglobin 
iron trapped in the area of the tumor becomes available 
for reutilisatlon by the host for the synthesis of hemoglobinj 
catalasa^ and the cytochrome systems* The studies presented 
above suggest that the rate may be very slow and that the 
iron lost into the tumor is relatively unavailable to the 
host in view of the short survival time found in animals 
with these tiimorso 

Pro^oaed cours e of projees : Ho further studies in this area 
^eprop^©d"i'or'''i'li© immeii'iate futureo 



48S 



Ssrlal Noo HCI^3l|l 

PHS«NIH 

Individual Project Report 

Calandas? Yeas? 1959 

Pa^t^» Honors? Awards* and Publications 

Pulvlications other thaua abstracts from this project: 

!«. Gjpesnfieilds. Ro E* » Sterling* W» Re 9 Prices Vo Bo s 
Studies on the anemia of tumor«bearing aniraalso 
IVa Distribution of radioiron following the 
extravaacular injection of labeled erythrocytesp 
Je Nato Cancer Insto ^i 196O0 

2o Greenfields R« Eb g Sterlings VU Ra » Tarantola* Vo Ac » 
and Price; Ve E» : Studies on the iron°>metabolism 
of turaor^bearing anlnials» Acta Unio Intern 
contra Cancrum Igs 888-892* 1959 » 



^sn 



Serial Noo NCI«353 
PHS^NIH lo Labo of Biochemistry 

Individual Project Repo3?t 2o Enzymes and Metabolism 

Section 
3o Bethesda^ Hdo 



Project titles The Chromatographic jiaalysls of 
the Hibonucleas© Activities in 
NoKaal and Neoplastic Tissueso 

Principal Investigator: Mary Eo Maver 

Other Investigators Antoinette Eo @reco 

Cooperating Unitss None 

Man Years (calendar year 1959 )s 
Totals l«l/2 
Professionals 1/2 
Others 1 

Project Description; 

Objectives To separate and characterize the multiple 
RNase activities in normal and neoplastic tissueso 

t4ethod.s employeds The tissues being studied are calf 
spleens rat lymphosarcoma* normal rat and calf liver 
and two transplantable rat hepatomas* Crude extracts 
and our more active nuclease preparations from these 
tissues are subjected to column chromatography » using 
first diethylarainoethylcellulose, followed by analysis 
of the RNase peaks on carboxymethylcelluloseo The 
RNase activities are characterized by their pH optima 
on highly polymerised rat liver RM* their products 
of hydrolysis of the cyclic purine smd pyrimidlne 
nucleotides and their heat stabilityo 

Ma.1or f indings; (1) The acid RNase activity of spleen 
has been "characterized heretofore by its ability to 
hydrolyze cyclic adenylic acid to 2« adenylic acido 
By re chromatography on carbosymethylcellulisij'i. the 
cyclic splitting activity could be separated from the 

SEE CONTINUATION SHEET (A) 

Part B included; Yes 



LlPc: 



Serial Noo NCI»353 

CONTINUATION SHEET (A) 

acid RMaa© acti-^'itjo The RNas© activities of rat 
l^phosarcoma tissues closely reseiobled those of calf 



(2) The acid RMase activities of the liver and hepatoma 

tissues hydroljze only cyclic pyriiaidin© nucleotides 

to give the 3 ' "^i^seleotides in contrast to the corresponding 

RNases of spleen and lymphosarcoma which hydrolyse both 

purine and pyrimidin© cyclic nucleotides to yield th® 

2* derivativeso 

Sifflificance to biomedical research and the program of 

the In stitute s 

Recent amino acid Incorporation studies with isotopes 
have emphasised the importance of RNA in protein 
synthesis and growtho Pancreatic RNase can Inhibit 
protein synthesis by acting on the RNAo Similarly^ the 
intracellular RNases may control protein synthesis in 
vivoo The use of these may also be of value i£i elucidating 
RNA structures 9 lo®o» the sequence of nucleotides within 
RNAo 

Proposed course ^^ of_ ^ the ,pro;^ect s A continuation of these 

studTes is' piaiimeS'"'aloHg" th® '" same lines » In order to make 

a quantitative and qualitative comparison of the RNase 
activities of normal and neoplastic tissues » chromatographic 
analyses of crude extracts of the tissues are needed sine® 
some RNase activity may have been lost during the chemical 
procedures used in obtaining the highly active nuclease 
preparationso Th© specific activities of the RNases will 
be tested on "core" and sin^ler polynucleotideso 



^86 



Serial Noo NC 1-353 

lo Labo of Biocheraistry 
2» Enz^es and I^etabolism 

Section 
3» Beth®sda« Mdo 
PHS-NIH 
Individual Projeafc Report 
Calendar Year 1959 

Part ^ Be Honors s Awards* and Publications 

Publications other than abstracts from this projects 

i:4avQr» Me Ee e Peterson^ Ei._ A'» ;. ■'3ober» Ho An » 

Grecoa A» En s Purification and characterisation 
of ribonuclease of calf spleene Annals of the 
New lork Academy of Sciences: 8l» Article 3» 
page a 599 ="610 » 1959 «» (!^his artTcle was in press 
at the time of th© 1958 annual reports) 



487 



M\ 



Serial Uoo NCI-359 
PHS-NIH lo Labo of Biochemistry 

Individual Preject Report 2« Enzymes and Metabolism 
Calendar Year 1959 Section 

3o Bethesda, Mdo 



Part Ao 



Project Title; Studies on the fractionation of 

e3?ythf'ocytes and other tissue cells 

in normal and tumor<=bearing animalSp 

Principal Investigators Robert Ee Greenfield 

Other Investigators: Mary Washk© and Betty Whitaker 

Cooperating Units s Dir'sioa of Biologic Standards, 
Labs of Blood and Blood Products 

Man Years ,CCalendsr yesr 1959) 
Total: |a 
Professionals 1/2 
Others 1-1/2 

Project Descriptions 

pb^^ectivess Part !«, Simon md Topper of NIAMD have 
'sH'eim' 'wits' th© use of radioiron and standard fragility 
techniques that older red cells are more fragile in 
hypotonic saline than young cells » Previous studies of 
our secti^^ have shewn that when the constituents of 
red cells of animals bearing lymphosarcoma R2788 or 
hepatoma 3^83 ^er© fractionated iis to their osmetic 
fragility by subjecting them to a continuous linear 
gradient of sodium chloride concentration ^ two populations 
of cells were obtained » One population had a maximum 
amount of hemolysis at 0^1^.% salt cencentration, which 
corresponds to th© peak found for the cells of the 
normal rat^ and the other was more resistant with a 
maximuia amount of hemolysis at 0o2^ salt concentration o 
Animals with lerge tumors and a high percentage of 
reticulocytes were found to have a high props^tion 
of red cells which were hemolyzed armiisd< 0o2^ salt 
concentratis) o By the use of radioiron^ it was found 
that these red cells which hemolyzed at the lower salt 
concentration were relatively young cells, but not 
necessarily reticulocyteso 

SEE CONTINUATION SHEET (A) 
Pert B includeds NO 



^88 



Serial No» NCI-359 
COKTINUATION SHEET (A) 

A simila? peak at 0o2^ salt con centra tlon was found on 
samples of blood obtained from rats five days after they 
had bled 50^ of their blood volume « 

One of the objects of the present study was to determine 
how closely the fragility of the red cells correlated with 
age in both populations of cells o 

Part 2o In the above experiment s^ the hemoglobin from 
those cells which heraolysed was continuously separated 
frora the remaining intact cells by a millipore filter in 
the outlet of the blood chamber which permitted the passage 
of hSKsglobin but not red cells Dr© Paul Schmidt of the 
Laboratory of Blood and Blood Products, Inato of Biolo 
Standards suggested that this same procedure might aid 
him in recovering and preparing red cells for injection 
after they had been frozen in glycerine It was decided 
to collaborate in the construction of an apparatus suitable 
for carrying out this separation o 

Pert 3o Is3 the experiments above (1) it was found that red 
cells which have been in the circulating blood more than 
twelve days hemolysed at a salt concentration around Ooij.^ 
vjhile less raatiare cells hemolysed at about 0q2% salt 
concentration o In view of the wide separation between these 
two populations of cells^ an atteiipt was made to hemolyze 
in a batch process all ©f the matxare red cells in the hope 
that the remaining immature cells would provide an estimate 
of the erythrepoiatic activity ©f the animal „ 

Methods employed ; Part lo Neimal rabbits were injected 
intraperitoneaify with a single dose of radioirono In this 
way, those ceils making hemoglobin during the brief period 
that the radiolron was available would be labeledo As mature 
red cells do not make hemoglobin or take up ironj, only the 
iramatiire cells would be labeledo ^t intervals over a period 
of two mcsjths, blood sanples were taken. The red cells of 
these saii^les were fractionally hemolyzed as to their osmotic 
fragility in an apparatus described in the 1958 report on 
this projecto The hemoglobin and radioiron released at the 
various salt concentrations were plotted for each sample o 
Thus, the age of the cell coiald be related to its fragilityo 

SEE CONTINUATION SHEET (B) 



489 



Serial Noo NCI^359 
CONTINUATION SHEET (B) 

If blood was tsjithdravra from rats five days after they had 
bson bled 50^ of their blood volmES^ approximately half of 
the red cells appeared in a population of cells which 
hemolyasd maximally at 0e2% salt concentration. At 
■various intervals before and after bleeding^ radioiron 
labeled cells of known age wiSE'e injected intravenously 
into these rats to serve as markers o By this technique 
it T-jas possible to determine in what cell population 
a cell ef a given age would be found » 

Part 2o An apparatus and purtgj was constructed by which 
red cells in glycerin could be circulated under pressure 
from one standard transfusion bag to another through channels 
beneath a sheet ©f milliper© filter papero The glycerin 
in the blood passed through the pores of the filter into 
collecting channel So 

Ma ijor findings; Part lo The nonaal erythrocytes of rabbits 
were found"to become progressively fragile with timeo 
Whereas, labeled young cells hemolyzed in the region of 
0o2% salt concentration J they progressively changed with 
age until the older cells hemolyzed at Oo5^ saline con- 
centration o 

Studies ©n bled rats indicated that the second population 
of cells which heanolyzed at a salt concentration of 0o2^ 
also become more fragile with agSo Immature red cells and 
normal reticulocytes hemolyzed at approximately Ool^ salineo 
After three days they hemolyzed at about 0^2% salt concentra- 
tion and after twelve days, they hemolyzed in the region 
of Oo3-0ol^.^ salt concentration J, thus entering the second 
cell population characteristic of mature cells o Studies 
in normal humans have indicated that a similar population 
of cells which hemolyze in the region ot Qo2.% salt con- 
centrati«:i arises within a few days following a phlebotomy 
of 500 mis of blood o 

Part 2o Although the apparatus designed to wash red cells 
free of glycerin is still in a developmental stage, it 
promises to becc3ime an efficient method,, This same equipment 
will provide a method for fractionating large qusrfcities of 
blood by fractional hemolysis for biochemical studies en cells 
of varying agQo 

SEE CONTINUATiai SHEET (C) 



490 



Serial Hoo NGI»359 
CX) HEINUATICK SHEET (C) 

Part 3o Tbs iinmatupe cells of less than twelve days 
of aga can b© separated from ttie mere mature cells if 
a gradieat of sodiism chloride approaching OoSi concentra- 
tim is sloT-ily added to blood over a 30 minute period o 
As yot^ when the ©rythropoisti© rats is normal or at low 
levels^ rather large quantities of blood are necessary 
to give sufficient cells so as to minimize the losses 
encountered during the wash steps before the final 
hemoglobin measurement o 

Signific ance I Part lo The demonstration of a close cerrela=. 
tien betweea age and fragility of mature normal cells may 
prove to be, an extremely useful tool in both fundamental 
and clinical studieso It will make possible a rather systematic 
study of enzyme changes which ace^apany aging of the cello 
It will gi-g-e a quick estimate ©f the state of erythropolesis 
in certain patients and may prove to be useful in the 
prognosis ©f patients in diseases with blood destruction o 
Although both abnoi^al sells and ©ells handled during 
transfusion have changes in fragility net directly related 
to the age-fregility relationships of noMial cellsp further 
studies may be able to utilize these changes to provide 
quantitative information as to the niaaber of abnormal cells 
in a given bleed sample as well as indicating by the change 
in fragility the extent v.^f ths oells abaopmal&feyo 

The deioen strati on of a close cors'elatien between age and 
fragility in immature cells is especially excitingo Although 
immature calls are found in the normal rat in only small 
nui?iberSp reticulocytes smd nucleated cells often account 
for over 90^ ©f the cells of certain rata with advanced 
tumerso 2?be ability ^©separata the constituents ©f 
cells which have nuclei frcsa those which have progressively 
less DM may provide another approach to the study ©f the 
relationship between DNA^ RUA, and protein synthesiso The 
ability t© fractionate the constituents of immature cells 
as to age will be useful in studies en the differentiation 
of red ©ells as changes in enzymes ©r proteins ©an be 
more exactly delineatedo 

SEE CONTINUATION SHEET (D) 



431 



Sspial Noc NC 1=359 
CONTEJUATION SHEET (D) 

Papt 2 and 3o The valu® and sigplficanca of these 
techniques cannot b® evaluated until further 
exp©E'i©ncQ demonstrates their accuracy and eflaiciencyo 

Pro^psed^ eouyse^^^ef ,, ;pgo.1ects Until the presentj, studies 
on the ^Factions from red "calls have been confined to 
the msasureiiient of aldolase^ catalase,a<?ult hemoglobin^, 
aud the uptake of iron and chromiumQ It is hoped that 
in th© cffining yeaPp studies will h3 initiated ©a othe^ 
cohstituents ©f red cells au^h as DM, RMg and Qiu 
uptake into proteins© In E,dditiegi^ t^sese investigations 
are planned to include studies of otiiiJ^p iiaraoglobin types 
with special emphasis ©a fb^«^l h&iyi^gliohiia,^ 

Mithin tb© past month a ne^a proeedurci has h^m ds^Fei^jped 
fop the ssparation of li^hole ©ells from the "11-^^v mid- 
tu52©3?o Initial studies indicate that this procedure will 
provide high yields of individual cellso It is hoped 
that in the coiaing year this technique of sd cell fractioni 
tion may be estendsd to cells ©f other tissues. 



i|92 



PHS-NiH 

Indi-^ldiaal Project; Report 

Calendar Xear 1959 



Serial Woo NCI-360 

!« Labo of Biochemistry 
2« Enzymes and i^etabolism 

Section 
3o Bethesdae Mo 



Project Titles Phosphodiesterases of Normal and 
Neoplastic Tissues* 

Principal Investjigator! Mary E» Haver 

Other Investigators Antoinette Eo Greco 

Cooperating Units s None 

Man Years (calendar year 1959 )s 
Totals 3/I4. 
Professionals l/l^. 
Others 1/2 

Project Descriptions 

ObjectJ -ges s To separate and characterize the 
^osphddie'sterase activities which are associated 
with the DNase and RNase activities of the tissueso 

Metho ds employed s The phosphodiesterase activities 
associated with the RNase and DNase fractions are 
assayed by (1) their hydrolysis of cyclic nucleotides 
and (2) their hydrolysis of calcium Ibis»(£nitrophenyl)- 
pho8phat@]2<> These assays are made on the phospho^ 
diesterase activities eluted when crude extracts of 
the tissues and our active nuclease preparations were 
subjected to chromatography on diethylaminoethyl cellulose 
followed by chromatographic analysis on carboxymethyl- 
cellulose* 

Ma j or , ^ f ^in^dindi^s s A phosphodiesterase activity of 
spleen wliich yields 2' adenylic acid upon hydrolysis 
of cyclic adenylic acid and does not hydrolyze RNAj 
is obtained when the acid RNase activity eluted from 
diethylaminoethyl cellulose is rechromatographed on 
carboxymethylcellulosOfl The acid RNase has lost its 
heretofore characterizing ability to hydrolyze cyclic 
adenylic acid« 

SEE CONTINUATION SHEET (A) 
Part B included s NO 



hSS 



i 



S©s?ial Noo NC 1-360 
CONTMUATION SHEET (A) 

The phosphodiesterase activity on the "bis" nitrophenyl 
eomponnd ascoapanies bofeh th® DNase and RNase fractions 
©lutad from diethylaminoethylcellialose columns » but 
no consistent correlation has been found between th® 
amount of nuclease aetiiFity and this phosphodiesterase 
activity* 

Si^nificM|e©s These phosphodiesterase acti-vities hydrolase 
phosphoHIestep bonds and may therefore be important agents 
in ribonucleotide synthesis which in turn is a factor in 
and protein synthesiso 



Progos^GourgeJ Studies will be continued along the same 
TIHesT^wTth special efforts to separate by chromatography 
these phosphodiesterase activities from the nuclease 
activities© 



431^ 



Serial Noo NCI -363 
PHS-NIH lo Labo of Blocheralstr-y 

Individual Project Report 2o Enz-^caoB and Metabolism 
Calendar Year 1959 Section 



3o Bethesdae Mdo 



Par-t Ab 



Project Title: Studies on the Cachexia of Turaor« 
bearing Aniraalso 

Principal Ini?6stigators: Miloslav Rechcigl» Jro 

Other Investigators: Flora Granthasa Robert Eo 

Greenfield 

Cooperating Units: Hone 

Man Years (calendar year 1959): 
Total: 2-1 A 
Professionals 3/lj. 
Other: 1-1/2 

Project Descriptions 

Objeetiyes: In the course of the studies concerning 
thee" t i olo gy of anemia in the tumor-bearing anitoals 
(Project N09 3i|.l) it was