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3. A third prnjected scak is to be hascd upnn animal skin tumnr prnducti<,n by an accclcratcd mclhtKl, if plant malcrialire. The relative evaluation of various "unknown" suhstanccs on these scalcs is cnntemplated in due course, when background infnrma- tion becomes adequate. Cnn/errnrr on Rlna..oy and Carr(noR.nr.l. Many of the matters outlined above were discussed in a round table conference D++ May 25. 1961, in the offices of the Committee. (letides memhers of the Scientific Advisory ilnard and staff, the following guest scienlists parlieipated: nR. T. 1) f)AY, tJniversity of I eedt Schnol of Medicine. 1 red., FnRland DR Wi1t 1 f R F HI'-CT(1N, Natinnal ('.ncrr Inailule, hrrttrada. Md t)R hRfl)t)Y FIOMRl1R(iFR, Rio Re.e.rch )nstitute, Inc ,('.rn- hridRe. M a+. F)R ROIIFRT W_ I-Illl l.. NmAwcstrrn llniversity, F.vanston, 111. F)R l-IIAR1 TS 1. KENSI.FR, Arthur I). I.iltle. Inc ,('amhridRe, M .at hR. WII I IAM E. P(1ft_, l)niver,i/y of PittthurRh, Pitt.hurRh, Pa. DR MURRAY I SIIFAR. Nalir,n.t Cancer Inttinrle, Rethetda. Md UR ('ONSTANTINfi S SIFP/IANO, Slrhhann Hrorhers, Inc Philadrtr,hia, P.. I)R I-IARO1.1) STEWART. Nalional Inatitutet nf Flealth, Relhr+da. Md MR O. F, lO1)I), Tobacco Manu/acturers' Standing ('ommillee. I ondon, FnRland. 11. Iltlmnn Lr.nR Studic+ Among studies on pulmonary function and physiology was a ciRnift- cant contribution in the measurement of "airway retittance" by the mcthexl of body pkthysmnRraphy. Inhalation of cigarette snroke both by normal pcrsnns and patients with various lung diseases, whether habitual tmnkers or nnnvnnkerc, produced a mild hrnnchnennstriction lasting from 1(1 to Rtl minutrs in mt,tl individual,. The effcct did not appear Itn mcdiate through nicntine nr other volatile suhslances aml crnrld F,e preverrted by ist,prrrtcrcnttl I)ata were Inadcquate tn warrant rclatinP of thit cllcct cautally In initfitir+n ca aRRravatinn of chronic pulmnnary diaeaa A rrvirw I,as .t,p-rarril tlrtcribin(- emf,crirnrr in Ilre crn,tsrr.,rivr alndy ,i1 (•ahuoe>fit al :vt:,tnmy r,f Ihe I,umin hrt,nchi:rl trce and lunpt whit h Ihe Scientific Advi.r,ry (tt,art) initiated in 1954. Alxtul .l,OINl catet were in- vettiPalcd, yicldinP R,Sv() alidcs from males and 3,661 fnnn femalca. Particip:,tinR wt•rc 12 fathok,Riatt in different Mcatiuns around Ihc cnuntry. 1lrc rcview rcf.r,rt, chictly nn prnMems of diaRnncis and clacsificaliun 'I he study it now hcinK extcndcd under auspices of the llnilcd Slates F uhlic f Ieahh Scrvice. A more tlct:rikd refxirt by one (d the participants in the cooperative ttudy has indicated that only 4R percent of the cases could he a+ncidercd normal and fewer nf Ihese occurred among smokers than among nnn- smnkerc Rcarltt of such sludies indicale the many prohlems in defining pathological chanRec and interpreting their significance. Ill. Cnr.liornscttlnr S/f1rltlPa Several nf thc carditrvatcular diaeates have hcen rcpnrted by invttti- Ratnrt in cpidcmi,drtPy In he asancialed with the use of tnf.accn, ctpccially cigarel/et As in the cate of the other asseuiatinn, these repnrts have given impctut to additinnal sludies on the cpideminlnRieal level as well as to many nf a clinical nr c.f+crimental nature. The object has been to determine whether the actnci:rtinns are truly meaninRful and whether they reflect any cnnlrihulinn by tmnkinR to the etiology of theae ditcaces, or whether the tink with snmkinR it only fnrtuilnus or indireet. IhrnuFh intcrmcdialc ct.m• mnn factnrs. i Mnny nrhrr Fncn.r. Al.o /f.InR Crur/lrr/ Among other L•tctnrt trnder investigation in studies of cartliovatcular discate dcvclnpmcnl are heredity, emotional constitution, slress and the individual manner of reaclinR to stress, physical activity in rclation to caloric inlakc, rclativc h<xly weipht, mclahnlie charactcrittict. Ihyrnid activity and dietary h;thitt, eepecially with respect to the amnunt antl kindc nf fat cnnsurncd Scx and ethnic origin are also involved. Recent prnprctt in several centcrs suRpests that a distinctinn may need In hc drawn Ixxtwccn IhtKe influences that hlfect the rale of presRrct- aiun nf chronic vawul:rr deFenerative prncetses such as alherntclernait, am1 Ihnte that may hclr prccipitale acute epiaxks auch as myncardial infarc. titm, thr mcchani.mt n( which still clmk full chrcitlatir+n. Vvidence rmrrtsinR frnm the Tnh;reca Inth/ttry Rcaearch ('nnlnlrllce prnhram and ekewhere supt-eett that the prevalent tyl,cs of chrr,nic v:,tt ular tkqlcncr:rtinn are nnl ai/•nific.rntly influcnccd in ralr or rlcPrcc by tAr I,criralrc n,lr.nni.tr:,rn,n .d nr, r,Ime in .m:rl) anu,unlt .,vcr :r L,nr• la r-d nr hy Ihc mtKlt'nrlt• I,r.rtUtc rd tntt,kinP f y avcr.rP,e Iti•r.,m. Slrll yd,arrrt• ur 16 17

Ihe mccli.rni.mc that l,tccirititc an acutc epittxlc in pertnns with advancc•d undcrivinF voccul.r tlitcase, who arc aontcquentty mnrc vulncrahlc It, all kindt nf ttrctt. F/huts Itt sr,lve these prohlcmt tit the cpiderninlr,Fical level arc fraufht with many dilficnltict Prearmahly the dcRree of v;rcc'ulir tlcRcnerntinn in human hc inFs can he accurately determined only at aulnpty by anatnmic nxthndt I hnuph aecidcntal deaths ennttantly prnvide tamplet which cut acrntt all linet of age, tex and race, the problem nf acquirinR the needed data on life hahits of thete victimt, by retrnsl+ective mcthtxlt, is formidable. Prntr%cctive audics. while permitting hetler accets to informatinn aMwt Lersnnality, dicl• smnking. Ma1y build and the like, do nnt cnneluce to actluisitinn tit anaU,mic information in a univcrsal nr nnn-tclcctive, randrnn ntanncr anrl thut dclx•nd u/wrn vaRucr clinical indicafir,nt nf cardit,vatcul:v L:rthut„Ry 1 hcy mu.t, mnrcnver. (h,r I,raetical rcatnm, u.c IM,I,nlatit,nt cnnfined to the nldcr age Ernurot. Ifmme,n Ar(rrlra Arr F.fnrnin.d 6 t)cspile difficulliet, a pathnk,Ric-an:)Inmie ttutly of human artrrict, rccnvcrcd frnm accirlcnt victims. it bring suropnrtcd Pertistcnt and in- peninus tflortt are mnde to acqnire and validate at mitch dala at IH,ttihle. retrntf,eclivcly, ahr,ul the pertinent charicteristict and lifc hahits of the suhjccts. ln enmpare irl+rnaches. techniqurs and findlnFt in this ficld• a cnn- frrencc wat held during the year to which scicnti.tc involved in studics involving timilar tcchniqucs end pruhlcmt were hroughl Inpcthcr. ('onfrrrnrr on Arhrroarlrrn.l. Crurfiro lhis cnnlerence wat held in the nrfices of the Tnhacen Industry Re- acarch (•ommittcc (in May 26. 1961. In addition tit s1aR and mcmhc•rt of the Scienlific Advitnry linard• the following invcttiRitnrs p,rticipatcd I1R W11 I IAM (7 PfAI)1-NKOrr', New York St.,tr hrp:,rtmrnt of Ilcatth, Alhsny, N. Y. I1R 1/1OA1AS R h.m, M.,.t IlAWf11 R• ll S Puhlie Ilc-1l)h Scrvice, I riminR- IrR IHA (iORli. vclcrans Adminktr:rlian Ilnapir.J- Wc%l Rn.hury, M.,a . anJ Ilarvard MeJical School 11R (i1 ()RIn (7A110 and IIR. MI('ItAI I I.YONS. New York - ity (Inivcrvtv ( nlttl:c of Mcdicinc• New Ytnk ( ON 1)R 111 NRN' (' tR , and MRS MYRA I RI('ItAR1)S. Mi(al l cn , 1 r„u.,..n., Ct.,/r Ilnr~rr.ity tth,~,l n/ Mril~i inc Nrw fhlr.,nt 1:, CJ © ItR (il ()Rt:1 V MANN, V.~n~1nlnh Ilnivrrvey C,hrwdof h1r,h,inr. m N,,.h~rllr, I rnn IR I)R 11'YKIf)IIR 11 RINZ1.IiR, New York llospital-('ornell Mcdical ( entcr. New Ynrk, N. Y. 11R WII HUR A. 111(IMAS, Albany Medical CoI1tRe, Alhany, N. Y. 11R. (il VR(il-. I WAKfRI.IN, American Fleart AtvKiatian, New York ( fty 1'roFrraa rn tirn.lirt n/ Tnhnrro E/j(.rh An cpidcmirdugical study on a large population of living male sub- jccls, including m.rny measurements and functional tcsls, provided no cvidence tit larl;c tit irnportanl dilferences in eirculalnry activity between gmup% of h:lhrtual .rutrkcrs and mrovaokers. 7 here wat little evidence uf Jctcrirmratiun of t:ardiuvascular '•filness•' in smokers performing work tests. Ilcuvy smrrkcrs had cnnsistently k+wer relative body weight, and higher L:rt:rl rr.y'cn crmvumptinn. Oilfetences in resling blood pressures were cithcr nnnaKnitic:mt tit tcndcd to he lower among smnkets. Itecau.c tit rcl,t,rtcd ditfererxes in the relative incidence of itchemic hc:vl tlist•:rse runnnl; the thfee major elhnic gnwps of Sewlh Africa, a stuJy t wat n+:tde :)mtmg (d1O healthy rcpresenlalives of these gruups, (•unsidcra- tirm w:t% rivcn lu trnukinK habilt, ineume, uccupalion, diel, anlhrupnmc(ric mcasurcrut•nlt, arterial pretsures, and total bkxxl cholesterol levelt, as wcll as di.trihulinn tit this lipid between alpha-and hrta-lilxrprotein. No .iRnitir.ult rclalionchip was faund hetwecn smoking and aRe, income, n:rlurc tit tacupation, hciRht, weight or degree of ohctity. A highly con- ti.lcnl dilfcrcncc was luund in the dittributinn of eho/esterol hclween Ihe :rll.ha-:md Ix•I:r-lil,trprnlcin (ractions, however, and these diflerences were pur:rllcled by dilicrcnces in the customary dielary fat intake. It was there- Inrc pnyartcQ thel snnding may e>tercite an Indirect in/lucnce by mtxlify- inl• the .clcctinn ol I(aKtt through elfccls nn the latte sente. Suhsidiary .IUJic% hrnvidiJ tnmc prclinrinary e.idence that smukers may prefer salty, sViced and hiFh-L•rt ItwKts and ntay esperience a diminithed sentitivily to hittcr t:r.tc. In a long tcrm prn.rc•ctive study detiRned to elucidate llre precursnrt rd hyrcrlcn.iun :tnrl ct-nmary disca.c, it has been reportcd that whitc nctle mctlic:tl atudcnt. whn arc the oR.prinR nf two parentt uffectt•tl with Ihese dr.c:t.c•t, :tre tignific:mtly diffcrenl from the ollsprinR tit Iwo unaliecled I+:rn•nt.. On the :rvrraRc• the tnnt nf the a0e.ted parentt are 1(1 httunds Fhcnvirr, m„tc frcqucnlly have hyl+crchrdetteremia, sh,rw higher tcttinR .ystulic hhMxl /ncarnc Icvels and we more likely to he smnkcrt. Whilc thcac dillcrcnce. :rrc nr,l yrcat, they ure eompalihle with thc hyla,thr.it r,/ multif:rctt,rial inhr-rit:,n,t• Ihc•ac r.htervalirrnt recall lite /N,tsil,ihly th:rt .nr„kinlt h.rlnt% m.tv rrtlrtt pre ctittinR cnnstitutinn:rl fat-Nnt. r:uhcr th.tn nccct.arily cnntrrhutrnr cauetlly In prrHlucinji, the diflerencet Io

I a, c r`. ,Stud-v o f Srrum Frr. Fntty Ar1d• The ohservation, from animal studies, that nicotine may rtlease cpinephrine and norepinephrine from depots and Ihus indirectly trigger the mtrhilitatian of free fatly acids (Ff:A) into Ihe citculation from tissue fat deposits, prompted a similar study in  limited number of human sub- jccts. "1 he smokinf of two cigarettes was found to produce an elevation in scrum free fatty acids. The response was not related to aRe, sex, Reneral smnkinR habits, type of cigarette or pre-smokinR level (if FFA. Total cholesterol was not affected. Non-smoking controls subjected to otherwiae identical procedures also experieneed a moderate but smaller increase in FFA. Such mohiliration of FFA was to have been exfxcted as a normal part of the resf+onse to any stress on the tympathctic nervous system. As such, it rctluires quantitative evaluatinn In relatinn In thc effects of all trther feums (if stress to determine whether such effects are of any real sipnificance in the atheroYclcrotic processes. Meanwhile, a different Rrnup of investigators has shown that in the pontahsnrptive state, the human as well as the dog heart extracts a cnn- sistently higher percentage of free olcic acid than of any other free fatty acid present in rterial hkxvd. It had been dcmonttrated prcviewely that energy prnduclirm of the heart tlei.rnde laryrly nn fatty ecid mNnholism. Animal studies of vascular I+athr.loyy, th(rur.h much mnre easily enn- Irolled, are impeded by problems of %f.rcies dillerrncrs and rtiflicultiea nf interprcintinn and aprlicatinn to human heinRt Ilrrwevcr, furthrr •tudics have been eonrplNed with rahhits fed cholcstertrl and cnlar,nsecd oil, nicotine, or a combination of all three, in comparison with cnntrnlc. A previous study had reported an apparently synergistic eflect of nicotine in spcedins and intensifying the production of peripheral circula- tory changes and cardiac damaRe, with necrosis, by the atheroRenit diel. In the present exf+eriment, however, the peripheral circulatory changes were minimal and cardiac necrosis absent. Since ergonovine had been used previously for lcsts to assess cardiac functional status, it was added to the reRimen, and the necrotie conditions were aRain obtained. These necrotic elfects are now attributed by the investigators to the addilional action (if erRomovine in the combination [Rccts formerly cnnsidered attrihutahle to the cholesterol diet, with or without nicotine, have diminished acctudinRly. Rr.pnn.r tn C7Rnrrrrr SmnRfnR by Cnronn" I'nNrnfa lhr rnv,K lrrrli,rl rrslwtntr Irv riParrttr tmnkint, in Iialirntt with ctrrr+nary fi,r:r" h;r~ I"n mrlsnrrrt hy tathrtcriratinn trchnhlucs 1'trvinut dudics h,d shnwn that smoking enhances myocardial blood flow in normal sub- jccrs. 1lcart rate was accelerated in both groups by smoking and :rrterial pressure generally rose, hul these changes tended to be more pronounced in the coronary group. A uniform increment of left ventricular work was also more pronounced in the lalter. Myocardial oxygen usaRe was virtually unchanged T he coronary subjects did not experience increased myeKardial blood flow during smokin6, perhaps on account of the "fixed coronary resistance" alleged to exist in such palients. There was no evidence of myocardial ischemia during smokin6, however, even in the coronary group. A study of DuerRcr's disease among young men in the Orient pro- duced evidence that this disease is distinct from the atherosclerosis or emholism ot the peripheral vasculature that occurs more commonly In the Unitcd Slates, and where actual existenee of a distinct fluerRer's syndrome has been qucsliuned. While all lhe 62 patients observed were smokcts. Ihe study suPRested that the etinlnRy and palhrgenesis (if the condition are probably complicated. Whether the essential ksion is truly an anRiitis re- mains to be proved. Pipe and ciR.ir smoking have been shown to prrxfuce circulatory effects (a drop in skin temperalure, increase in pulse rate and hlrxxl prcssure) similar to but less marked than those found following cigarette smoking or tobacco chewing. IlallislrxardioRraphie changes were less than those follnwinR chcwinR, hut greater than those following cigarettes. In an effort to throw light on the physiolo6y of relative hyperrcactivity to smoking by certain healthy young adults, in terms of blood pressure, heart rate, or cardiac outpul, the effects of preliminary small intravenous drxts of hexamethnnium, mephentermine or of a talt-solution'placcho wcre studied. lndividual differences in homeostasis were revealed but the changes were not readily grouped and the resulls were too varied for sirnple classification of suhjects. I Fundnm.nln/ Stndtrs of Cnrdlmvr.rufnr Cyarm Numenws contributions to study of the cardiovascular system alonR hmdamental lincs were made during the year by recipients (if 'lohaccn Industry Research Committee support without immediale bearing uF+r.n the eflects of nicotine or smoking. OIhers eonarihuled new or inq.rttvrd techniques for study (if this system. A notable series of such paf+ers centered auormd the mNaholic nc- livitics (if the hrart mutcle unrler varinus ennditirrns antl inflnrntcs anJ of Ihc rclatinm t,ctwrrn hir~chemicat events and druy, actir,ns, anrl tlrr resulting mechanitil and trthcr functit.nt. Othere dcalt with thr rntirrnatr 21

td rrrrtincntc -,a kractl urrrn hirxhcnrical ct+nciJcratinna, anrl wirh mrth+xlc t+l nrc. mint tmrnr:ny I.hxxl Ilr+w antl td :rcacaainl ttar+n.rry :rrlcry imnflrt rcnt y A t+t't•inninl- Ir.rc :datr I+ccn madt• :rt Ihc crdlrnc td nt+rnr.d ral hih' :rnrlic anJ rnrK'arrfi,rl ttll, in rirnr. Variunc nuLricnt rncrlia lravc Ix'cn lcatcd :rntl Ihc rcl•rtivc frr,wIh Ix+lcnti:rk t+f acvcr:rl ccll lyl+cc tlclcrnrincJ l)hjrclivcc arr tr+ nl.acrvc mcl.tht+lic activitict -It they nray I+c affct:tctt by a:rnt+ua c+t- U.rnct,uc auhaf:rncca, including nict+linc. 'I hccc cnntrihulitrnc atc imrtttlant ccicntific:tlly In thc furlhcr clutly r+f c:rrtlitrv:racnl.rr dicc:tcct and ahtlr:rclc of all auch p:Ilx•rt are inchulctl in Ihit rclxal IV. Ira~fllfr-I'Ir3.rrrlnRirfrf Sfrrrlir. 1 hr 1•cncr:rl t+hjrctivcc t+f rcccaft II in Ihic ficlJ have ht•cn dret uaarJ in Ixcvinrr\ rclx+ttc, :rntl 3 ctyntinninC rrr+rr:rm ic in thc f.rtat•cc nf e.l•:mainn ('utrcnt rcfx+rlt includc a atutty nf rcl:rtit+nahiM hctwccn ctmctilutitrn antl t+thcr ch:+rrclcriaticc nnd ainAinl: in a Frt+np of cuhjcctc all td Italian famity tail-inc frt+nr thc :rrc:r ncar Nal+lct. I can men were ftnrnd tty crnnkc nuuc 1h:rn att+ul nncc, hrrt thc tlictt wcrc nt+t cipnificanlly tliffctcnt Slit•Itlly hir~hcr ccrmn cht+lcatcrtd w:rc ft+untl amnnp amrrkcrc, hut cipnrficanl dif- fcrcnccc wcrc nr+t t+hccrvcd Ix•twccn trntrkcrc :rnJ nnnanrtrkcrc in anlhrtr• p+mctric ntciclrrcnrcnl, in phycic:d :rclivity or in :dcuhtrl t:r+ncnmf.lirtn. An anal,ycic uf thc amt+kinp frncliccc nf n rrrnrp td Il:rrv:vd nrt•n in crnn(,ariann wilh tht+cc t+f the pcncr:rl f+tq.nl:rlirm h:rt alctr nfqvcarrtl, ac a hy- f+rtxlucl nf a nn+rt• intt'ncivc ttutly t+f arunkine wc tcl:ltt•d ttr Mxly huiltl V. T.rlrnr•r•n CIeenti.rrv rtrtrl Mfrrlrentiarv A tcricc td alndict tin rncchinicrnc of hitrcynthctic of Nio-nritrrrn alka- IniJc hac rcichcd f+nhlicntit+n at culminatinn t+f a Inng-ct+nlinucd prt+Fram trt aludy Mcthanicmc t+f m:rnun:rli:tn and h:tclctinl nrctahrditrn n( nit'ntinc and relntcd :rlk aIt+irlt have 1.ccn Iraccd thrnuFh arldititrnal st:rgcc wilh full irlcnlificntit+n t+f inlcrmcdiatct hy exaclinR mcthrxlc. Scvcral new ctm- cliturnla of tnh:uctt have hecn itlcntificd. Scvcr:rl ycntc :rpn the qucctiun w-vt riicctl whclhcr the fr+rmrr uac r+f :rracnic:rl inacclit'idt•t mirhl have cnnt:rminalcd Ihc cr+ilc ul ItJ.accn prtrwinF arr,ta with cr+ncrqncnt hmF aualainctl int'rcacc in arct•nic ahcv,rf+tit+n hy I-+h:tccrt plantc A tllrrc-yc:rr atudy t+f thia auhirct, rcfx+rlctl prc'linualy, ah++wct1 Ih.rt cuth a+il ct+nt:rmin:ttinn Irm. nt+l txcurrt•d Irr any cirnilit:rnt Orrrcc A n. w it t-rt i+n rnrth-lr+lr-py nf acmi nrlt rn nn:rlytic l+n arat rnic ...r,~,~, . n.... I,u1~l~•1„ J will Lr~ rhl.~lr +ny Iu1~nr V1. (Ifher Strriflirs (11hcr cludiet include reports frnm the conlipuinR invetlipalitm t+f nicrdinc ~h:rrmaculn~y which cmphasi7cs cspecially cffcclc upt,n Ihc ccntr:d nerw+oc syalcnt with an ultimale view to elucidalinR snme of the bacct ftur the uae nf ttrh:rcco by humant in the cnntext uf tire silualinnc Tt+h:rcco uae hat been found Itr have no inffuencc on the tucurrcnce (rf criicrxlcs in tn-callcd "periodic" diseate. A notable event in 1461 was publication of the comprchentivc mr+ntr- Rraph. Tnhmrrr, r,rrerimcnrul and (7inir'af SwAiir, by I)rs. 1,rsnn, IIaaR and Silvette of the Mcdical Collcge of Virginia. This R(K)-paRc Mx+k, whnce cnmplclinn wac atsitted by the Commillee; shnuhl greatly increice the accectihilily nf tcientifrc literature in the field and lhus slimulatc and f:rcililate new reccarrh, while conlrihulinR significantly tt+ Ihe dcvchqrrncnl trf hetter perapt•ttivet 'I'he Ix><rk Is a camprehensive accormt trf the wurlJ li/rrature, I+astd on uudy of mtxe than 6,((N) arlicles publiahctl in ahtrul 1,2(N) jtnrrn:rlc. It hat been widcly and favorahly reviewed. Other comprehcntive reviews of Ihe literature on the eRectc of tobacco on sl+ccific rxfans or syttems appeared durinR the year. Thc litcrature an IunR cancer from 1930 to 1960, wat reviewed by Rirdnn and Kirchtrfl, who prcvioucly had puhliched a review of the freritxf 19(NI 1930. llrc laler arlicle, containing 455 refcrences, noted that varinut facttrrc have fxen chrdied in the etiology of IunR cancer, including tuherculosit, influcnra and other chett diccatec, nccurational e.pocures, atmncpfrcric pnllulanlt, anJ tobacco. It concluded that the cause of humnn hmR cancer it unknown. Afrdlrnf Ctrnf.nr Frfrorenhrpse Since IUSS the Tobacco Induetry Reteatch Committee hat, upnn rccnmmcndatit+n of the Scientific Advitt+ry Rnard, been cupptalinR a prtr- Fr:rfn dctigncd to ttimulate inlcrctt in hacie research among mcdic:rl tchtwd sludentt. Thic prt+gram wat again in eReet durinR the year among Ilu n7litm'c acercdited mcdical and ottenpalhie «htx.lc '1he imfxrrlancc t+f such a prnFram was recnpnired in the April 1961 rt•pttrl nf the 1'rcaitlcnt's (•rrnference on Ifearl hiceace and (•ancrr. lhit rcf+trrl caid that h:rcic rcccnrch muct be tuppnrtcJ at the chicf ctnncc nf ncw knnwlcrlpe and rn/'cd Ihal mnre ynung pettrlc nf nutet:rndinh ahihty t+c rccruilcd into the ht•allh fieW. ltndt•r the (cllt+wchip prtrgrmm ol the T I R(•., alnttrnlt. in ct+nculla• liun witlr thcir :rJvi.ura, trlcct the suhjcclc Ihey wiah t,t attrdy Uc:ma nf Ihr «'htxda t'hrx+ac Ihr fclh+wa, antl Ihc wrnk ic th+nc tlurinr cunrn+cr r+r r+lhrr r+ll Itrrm limt c umlr r Ihr aul+crvicit+n nf cclx•ricnccd rrat•archrrt In Ithr I:+•t tcvcn vr•rra ttrmc tlr:rn S(NI yt+nnrI+crat'n. h:rvc rrccivcrl Irllnw.hiln + Ilcrt'+I Ly 1hc 1,,,ticntific Advr.my Il++artl. 21

Abst racts of Rcports Fach recipient of a Tobacco Industry Rescarch Committee Frant-in- aid is rc~hnncihlc for the initial f+rcccntatirm or publication of the results of his rr%carch in ccientific mectings or in appropriate scientific jnurnals. FotlnwinR are ahstracts, approved by the authnrs, of rescarch rcpnrts, with acknowlcdRcment of support from the T LR C., that have appeared in scientific journals since the last Report of the Scientific 1)irrctor and thrrnrRh 1)cceml.er 1961. The name of the scientist tn whom the T.1,R C. grant was made is given in parenthcscs where required. ('rtpirs of ah- stracts in previous Rcpcsrts are avaiFahle upon rrqucst. -1 hese ahttracts have been grouped under six headings: I Cancer Rescarch; II. Ilurnan LunRStudics:lll ('ardiovaccutar Stutlicc; IV 1'%ychr.- phycir 11rFiC.l1 Slutllr4; V. Tobacco Chcmidry and Itita-hcmiatry; artd VI. ( )thcr Studics I. Cancer Re.rr:rch "Tl1MOR-!IOST RfF-ATIONSIIIPS STl1D1Fh 1N VITRO FXPFiRI- MHN 1 S WI I1I ( F-I.L-FRfiE SYS I IiMS." R~' Leopold R. C'erecedo, Sister Marian loce Smith, and lesus Vicente. Fkrearrment of (liochemiatry. Fordham l)niversity, New York City, and 1)cpartntent of Ilicxhemistry and Nutrition, School of Mcdicine. University of Puerto Rico, San luan. Archrvcs of Rinthrrnirrry and Rirrrhvsics. Vol. 92, pages 4R4-4R6, March 1961. (T.I RC. grantee: Cereccdo) Purlro., of .ru,fy: Previous shrdics by the authnrt have shown that in tumor-heannR rats there is an increase in the denxyrilNonuclcic acid (1)NA ) concentration of certain tissues at a distance fron, the tumor. l hese enects of the tumor could also he shnwn in virrn whcn the incorporation of thymidine Fl' and adcnine Cr' into the DNA in slices from lumnr-hearins animals was compared with that in the tissues nf control animilc. It was decided to investigate whether the stimulatury effect of the tunwr could also be shown in cell-free systems. Pror.dnrr: The incorporation of thymidine-llt into DNA was stuilied in high-speed supernatants of extracts of IunR, kidney, livcr, and spleen of normal rats and of rats bearinR the Walker 256 carcinoma and the Jensen sarcnma. FYndinR.: An increased incorporation was nhcerved in the w(+ernatants of the liver and Ihc spleen nf the tumrn hcarinR hntle as cnrnpatrd with that in the mrrmal cnntrota. The eflect of the lensen lurnnr trn the incorpnratitrn was more rirnnuuneed than that of the Walker carcinrnna -11iia inuca%ed activity was slin Irrund when the incorPtrratitrn of the IhymiJinc 11' was studicd in thc ~nrcrnatant fractirrna rrl thc turnru, Ihcm.ctvcv It ia travtrn- ahle Ill a«urnr 0111 t lnme factnr rrtwlnced by thr tumra an l tran~mirted tn 1hr.r ti.~uct hy way of thc thliw.l it involved in thit l+henr+mrnrm (Ithrr Rrn..rc.rar t/ S F'uhlic Itcalth Scrvice and the I)imrm Rtrnyon ~ Atrni "n:rl I irnd "TIIE F:FFECT oF SMOKE AND NON-SMOKE GASES ON CELIS IN ( Ii1.L ANI) ORGAN CULI URES." By Philip Crnper, M.1)., FACS, and Irene P. C7uldrinR, Ph.D., Department of SurRery. Albcri f:instcin Col- IeRe of Medicine, and Drnn>c Vetenns Administration Ilcnpital, New York. SurRirof Fnrum, Vol. 11, pages 72-73, 1960. (T.I.R.C. Rrantee: Cooper) Purpose o/ .rurly: Studies have already been accomplished on the pcn- sihlc effect of cigarette smoke on luns, in vivo, or on tissue slices, in vitro. The present study was initiated to investigate the effect of various 6aseous agents on cells at a cellular level. 1'roc.Aur..: Smoke from cigarettes, cigarette paper, or onionskin paper was injected into mammalian cell cultures. Controls included similar cul- lures exposed to atmospheric Eases, carbon monoxide, illuminating Ras, and other nnn-smnke gases. The gases were Injected sfter a growth period of 24 hours into Ilasks containing an original inoculum of 1(K1,0()p cells in 2 cc. of NCTC-109 synthetic media to which 10 percent horse serum had been added. Three cell volumes (0.3, 1, or 1.5 cc) of smoke were injected; total cell count of each culture was made after an additional 411 hours. Daily microscopic observations were made. ('ells in a Rose perfusion chamber were observed directly after the injection of cigarette or cigarette paper smoke. Cigarette smoke was alco injected into embryonic rabbit lung organ cultures. Lung organ cultures of I R day old rabbit embryos. grown for 24 hours in synlhetic media, were exposed to 5 cc. of cigarette smoke, and sections were prepared (rotn the tissue after an additional 411 hours. F7ndinR.: All smoke aases tested altered the growth curve of the cultures significantly, and a reduced cell population resulted. The chnnRes induced by cigarette paper or onionskin paper smoke were more pronounced than that produced by cigarette smoke. Non-smoke Sases tested showed no sig- nificant effect. Cells 6rown in a perfusion chamber became enlarFed, showed surface "hlchhinR', and increased eytoplasmk granularity and vacuolation. The embryonic lung organ cultures showed arrest nf growth and deRenera- tive changes in the tissue after exposure to cigarette smoke. l he effect nf the smoke gases, over prolon ged peri<x1s, may conceivably prrKluce chronic inflammatory changes in the lung. "A PRnTOT.nAN ASSAY FOR CARCINOGENIC FIYI)R(X'ARIIONS." Fly Robert W. Ilull, Ph D., Department of RiohMieal Science, Ntathwestern ttnivercity, Evamton. Ill. 1'rrrrrrAinRt, lnrrrnolirrnnl ('nnfirrnrc n/ I'ru- rntnalnRy. PraRue, 0eehnslovakia, Aug. 24. 1961. 1'urfro.r of .rudy: Although the hasie mechanisms nt phnlrKtynamic ac- tinn are still unknrrwn, e.ploitation o( the phennmennn of realamee ri/ PnHrrytra tu ultraviolet and related radiation for use in the dctcctirrn rrf hydrruarhuns in high dilution can have considerable utility in Ihe arca nl cancer research. 1'rerr.lur.: Srttutitms ttf hydrrKerMtns in water are prepared by either rrf two nlethlNh, I fle crllatet in an amount of Rlass-disliikd water cqual ur the v.dume of hydnK arhnn srrlutir.n are addcd to PyreR al.rrt Iest lst.rtrs or slanrlard alitks comtaininR the smrenaiuna. The eitiates are cunccntr:rtcJ so as to provide helwcen 20 and 30 ciliates per dcpressirrn Ihe.cilcrtr% 25 74

tnutinrly uaed are 1'aramrcitrm caurlarum Rrown in axenic mctlia or in mcdia cnntaininR Arrnhnrrrr arr.rRinit- Cr+ntrnlc in the IeN arc ciliafcs hamlled in eRactlv the same manner except thal the hydn+c:uhnn it vmittcd. The crliitcs in the hydrncarFxm susf,enait.ns arc held in total d.,rkncss for one hour or more and then expcned to liRht of known qualily and intcnsity. The time intcrval. between onset of expnure and imrnr+hihty nf W) percent of the ciliatcs is recorded. The effects (if varinus known light wave lengths between the maximum hacteriocidal wavc-IenRth and vicible light were examined. F'rndinR.: T he work done to date dcals with .landardisalion of the assay techni(ue as regards ils I) reproducihilily and range of sensitivity, 2) the efTecl of the quality of the irradiating IiRht, 3) the eflect of various intervals of exfanure of the ciliates to the test comf+ounds prior to irradiatir+n, and 4) variations in scnsitivily to a series of diRerent hydrocarbons, some of which are estahli%hcd carcinnRens. In Reneral, hydrocarbons Ihat are knc+wn front mammal tests to he carcinnfenic show a shorter kill tirne in the a.1 ay system than do hydrncarla+nt whose carcinoRcnicity has m+l been cstahfi.hrJ 7 his aaay prrdrels that certain of lhese ct+n,rounds Ic.tcd will be (r+unJ to be Nfective carcinogens. It can he concluded from Iheflala now al hand that this assay systcm is sensitive and reliable In the dc(cctinn of polycyclic hydrocarbons in high dilution It can he anticipated that (urthcr characlerisatirm of the tesl may provide accurate prediction of carcinnRenk potency of the hydrncarbnns. -lhere is alsn the very real(x ossibility that this assay system may provide a means for direct atlack on the hasic ntechanisms of chemical carcinnRencsis. "pOANTIT'nTIVf? ('Y7(X'HE?MICAL ST Ul)IHS ON T1IF RF1.nT1ON OF I)liOXYR111ONUCI.FI(' ACID Uf' Cf•LI.S TO VARIU(IS 1'A'111O- L(X71('Al. (Y)NI)I f1ONS.° fly Cecilic Lcnchtenhcrger, Ph I)., and Rudoll l.cuchtenherger, M.1)., The Children'. Cancer Rcscarch fr+und:rtir.n, Rnstnn, Mass Rir+chrmlral Phormor.>1nRy, Vol. 4: paRes I2R-IG1. Augud 1060. (T.I R.C. grantee: Cccilie LcuchtenhcrRer) F'nrpo.e of arrulv: The possibility of  direct correlation between the functional staRe of a cell, be it normal or ahnormal, its micnxc++pic appear- ance and its chemical composilion assumes particular significance in view of the fact that a change in function and mnrpholo F•y of a cell is nu+re or less the e.pressiim of underlying and often preceJinR chemical alteration. Simultaneous morphological and chemical investigation at the tinFle-ccll level miRht not only throw light on etiology and pathnRcnesis Mrt akn might help greally In the detection and understanding af discase. In particular, special attention In the study of dcoxyriFxrnucleic acid ( I)NA ) in ahnnrmal cells and in cells under pathologic conditinns suemal nreritcd in view uf the important role of DNA fnr cell life nd cell continuity, ita quinlilativc stahility under nr+rmal conditions, and il+ chne rtlatinn%hip tn thc Rcncs Pr..ridur..r I)urinR the Ia.t 10 years the quantitative hehavior nf DNA in cells has been rxtenslvely tnvestiyated by micrnal.eclnq+h++u+mNry >•nd inlerferrnce rnirrr,.++q+y in relali+m to fnur ralhul++piral I+rrKr.,ca, ( I 1 a+ngi+ nt .ttcss, ( 2) tumnr., (1) vir+rl ir+fcclu+n1, nd (4) infrrt hly In ttndym~ ( 1), &'ps were sul+icclcd /n pa.trcct++my and th++ltcy~lctturny, and cells nf Ilvcr and adrenals were analysed for DNA canlcnl In flte case r+f (2), hun,an precancerous and tumor tissues were studicd and c+Hrtrastcd with nnrntal hum:m tissue. Under (3) different typesof hurnan and other cells infected with diRcrent UNA-containinR viruses wcrc examined in iirrr in the host and in lissue cuhures, and in (4) more than .12,(NN) sperm cclls from over 2(NI men and from over 100 bulls were analyred for DNA cnnlenl. F'inr!inR..: ( I ) Analysis of more than 7,0()0 inrlividual cells from livers and adrenals front 17 dogs showed that DNA content remained unchangcd ofter surRical stress, although a considerable elevation of lyrosine was nntcd in liver cells and in cells of the adrenal medulla after surgery. (2) It was found that all normal tissues, no matter what their origin r.r nretahulic functinn, contained cells with a similar basic mean DNA crmtenl, whereas the DNA content of precancerous lesions and utali6nant tumors was found to be increased and revealed a much larger scatter from cell to cell. T tre deviating DNA data cannot he considered a speci(ic criterion for malignant Iran.fnrmatinn of cells, hu1 may be explained mainly on the basis of growth and miltNic ltrncrsscs present in most tumora. On the other hand, cnn-lancy of DNA in nnrmal tissues indkales that an Increase and large scattcr uf DNA in such tissuc must be looked upon with suspkiom of mnlignancy, un- Icss regeneration is trt be expected. BeniRn cells disclosed essentially a normal constant DNA quantity. (3) Study of different types of cells infected with different 1)NA-cnnlaining viruses in sirH in the host and in tissue cultures revealed a peculiar combination of intranuckar Warre conRlnrneration of DNA masses associated with striking increase and variabilily n( DNA 1 his almnst specific response of cells to an Infection with a UNA-containinR virus was noted irres(~cctive of Ihe type of such virus (vcnnca vulRaris, adcnovirus, efc.), of Ihe species and types of cells Infected (hrrman, insccl, nrxmal, or cancer cell), or whether the cells were still in .rirn nr explanted in tissue culture. (4) In contrast to the remarkably constant rind unifnnn haploid DNA content in s(xrm from fertile haman males and hulls, the DNA content in the slxrm from infertik males was frequently variable aml significantly lower than that from fertile ones. In addition to the various types of human tumors studied, the relation of DNA to sprmlaneous and transplanted animal tumors was alsn investi- ~,ted. Principally there were no essential diRerences between the DNA hchavior in huntan and in animal lumors studied. This similarity of DNA res(xarse deserves special cnnsideralion, because some of the animal tumnrs s rdied were tumnrs with a known vital etioloRy, namely Rnus sarcnma of c chicken and F'ulyoma tumors of mice. OIRr r Rrnntor.: (I. S Public Health Servke, Elnnh Foundation, I:ha F 1'ardee Foundatinn, and Franchester fertilily Fund of Clcveland, 0 "A ('ORRI:I.A 11:f) IIIS"IO1 (X31CAL, CYTOI.(Kil('A1. ANI) ('Y 10 ('Il1iMI('A1. ST(tl)Y OF '111E 1RA('f1EUHRON('IIIA1. I RI1E ANI) 1.(/N(iS OF MI('1: F.XPOS(il) TO ('1(iARfil-1E SMOKI?" Ily Cecihc I cuchtenl+erRcr, Ph UY and Rudolf l,euchtcnherger, M 1) , In.litul (iir nllFcmcinc Rntnnik, 1? I/1, 7l)rich, SwiVerlantl, .Irru M.diq a.f+,rndinnvu a. V+dumc 17/1• Sul.l+lcmrnt 1(i9, pages I(/2-11R, 1961. On the ha~ia ++f thc dala prcwnlcd sn far, it appenrs that cxl+nsurc t+ cirarclle sm++ke will evt+ke altcratir+ns in Ihe branchi r+l sume mitc xutu- R, 2~+ 27 rn ,r.

marired as follrrws: I. No significant histoloRical chanFcs. 2. Mild bronchitis; swollen epithelium frequently associated with mild hyixrplasia. 3. Severe bronchitis and (.erihrnnchitis; atypical hasal cell hyperplasia, squarnr+us cell meraplasia and occasionally histopathnlogical chanRes consistent with the terms "dysptatia" or "carcinoma in .tiru," One or more of the above changes may be observed in Ihe same animal. Although, on the evidence of these findings, one may he tcmpted to conclude that there is a dircct cause-effect relationship between cigarette smoke and thc epithelial ksions, there are mainly two findings in our study which impose caulion against drawing such a conclusion: 1. A striking difference of response is encountered front mouse to mouse which occurs regardless of whether mice were exfxxcd tn relatively low doses of cigarette smoke for a brief period or a relatively high dose of cigarette smoke for a long period. In each of the groups, over S(/ percent of the mice did not shnw any bronchial ksions, while 25 percent showed mild and 20 percent dycplastic lesiona. 2 1 here it absence of  rrlatinnship between dose nd length of ex- T,surr to cigarette imoke and frequency nd severity of lesinns produced. is evident that, in spite of the cnnsiderahle increase in length ol exl+nsure and numher of ciRarettes, the rehtirmthip between mice showing no siR- nificant chanRes nncf mice showing mild and severe chanPes in the hrnnchial tree did not alter rpreciahly. 11 it indeed astonishing that, among the last Rroup, no inva.ivae hronchoRenic ca+cinnma has been observed although these mice had been esposed to enormous quantities of cip,arclte smnke. and, whcnever t+.lstihte, fnr nearly their whole life span. The widc spectrum of findinRt and the independence nf rlusr and dura- tion of etpe,cure to cigarette smrrke would « rrn to detract fnmi the im- prrrtance of ciRarclte smr ke as an injurious agent invari,hly aflectinR the major hrnnchi and would, ra1her, point to contributing factnrs characteristic for the individual animal. Amon the many Irrx.ihle host factrrrs, vinrses descrve special cnnsideralion - t~e mnre so since they occur with a certain frequency in mice, sometimes in a latent Grrm, and are capable of producing respiratory Ietinns and proliferation of bronchial epithelinm. There are two main lines which we are investigating at present. The first is concerned with the detection of latent viruses in mice prior to e.- rosure to cigarette smoke. The second is eoncerned with the effrct of ex- t+ttsure in virut in addition to the exposure to ciFarettc smoke. "('OMMON IIIIMAN VIRUSES AS CARCINOGF ' N Vf ("1ORS." fly ('hridnpher M. Marlin, M./).; Sigmundur MaRnuttnn, Feiluw of the Squibb Inslitute for Medical Research Foundatinn; Philip ). (io%cicntki and (ier:rlJ 1: Il:rnsen, National Inslitutcs nl Ilealth rrsc:rrch fcll-s; Sclun Ilall (-nllcge of Medicine, Jersey City. N. 1. Srirnrr, 1)cc 15. 1961. paRrt IvRS-Ir)R6 (T I R C. granlce: Martin) 1'rr.p.r.r o/ .rrrrlyr It hat been a continuing paradna that carcimogcns ttrrrngly imf+licatrJ in hurnan Iumorir.enetis, rhr+ugh Irrrsrnl in thr hrnnin rnvirmnmrnr in nrdy trace amr-unts, nrdinnrily wFfl indurr nrr-lrlaaia In ;rnrm.rli r,nly whcn 1Htn11n1Urrc1l in relativcly largi• 111714111111% r1r wrlh vHrnrat t+hyak:rl rir t hr•rnicil "crr carcinrrRrnt " Spcculatinn rrn pocaihk natural nI cn-carcinngens led to consideration of the possible role of conrrnon, non- tumor vinnes. Studies were performed in vivo on 290 male Swiv white mice front a colony proved free of polyoma virus and which were reported to have a low incidence of de nnvo tumtxs. Pairs of known chemical car- cinogens and common viruxs in different combinations were administered by injection. The viruses used (vaccinia. ECIIO 9, Coltsackie Ilr and Poliovinrs 2) were harvested fiuids of fully infected tissue cultures of mnnkty kidney; by the routes siven, they evoked negligible mortality or ntorbidity. The carcinogens injccted and their respective drtscs, judged to he lon small to induce lumors, were (00 micrograms each of V,10- dimethylbcn?anthracenc-1,2 (l)MDA) or of 2-aminofluorene (A.F), anJ 75 nsicrograms of 1,2;S,6-dibenzanthracene (DRA). To determine the role of immunization, half of a group of IOR ad- dititinal mice were immunired against vaccinia virus nd half against fRozen-Ihawed monkey kidney cells prinr to administration of cither vac- cinia virus or frnzrn-thawed monkey kidney cells plus either 1)MItA or propylene glycol. F'inair,R.: Malignant tumors occurred in 5 aroups of mice that received crrcinrrRen-virus Pain, and in no other Rroup s. T'here were S maliPnant lumort each among 16 mice injected with bMIIA and vaccinia virus, y of which were living at 3 nronths, and among 12 mice injected with I)M11A and Poliuvinn 2. 7 of which survived to 3 months. There were 2/umars each among 12 mice injected with AF and F-CHO 9 virus, with 7 living at 3 mrrnths, and amonR 12 mice injected with AF and Cor<sackic R, virus, of which 6 survived to 3 months. Four Ihymomas - turnnrs known to arise de ne+vn in this mouse strain - occurred in the 161 mice alive after .l monlhs. After R montht, the only tumors ohserved in the immunizalion era- juriment mice were 4 lymphomas in a group of R mice that were not rmmune to vaccinia virus and were given vaccinia and hM11A intra- l•critoneally. in vinu studies! ('areinngen-hinding by Polinvirus 2- arrrr'xim-tcly ltl (NN) mrrlecules per virnr tissue cultorc. 50% of infcctiuus dr,cc - wat dcnwnslrated in rirrn, with (he use of I)MRA-9-('r41. The re-tuhs aflirnr in vivn interactions of viruses and carcinogens previously Jctcrit.ed by other investigaton, and are consistent with tlrc rc(+rut I+y 1) V. Wi.clcy ct :rl. (Proc. A A('.R. J:27R, I')(rt ) uf cnhancvr1 chemical carcinnPencsis in mice repeatedly exposed lo respirnNny viru.rs. If such interaclinnt occur in nnture, it may prove prssihle lu rrrlucr ne•rr- plasia currcntly ascril.c(l to chcmicnl carcinogens by inununizatirm nhain.t a Vlrut. "VIIANGfS IN Mt)f)A1. VAt I/F.S OF ('IIROMOSOMFS AFII R I/t RAt)IATION OF IlI/MAN AMNION (7it 1.5." ny Mat,hirrr t\lirut:rni, 11 A, and Y 1I N.rkanidri, Ph.1), now ot IhrkkaiArr I/nivrrsity, lutran; Y:+svJu l)frnuk i. M S, nrrw •t Pasadena, ('aI , I oundatirrn fnr Mr•rhr :rl Rcsearc'h, and (' M 1'rancrat, Ph 1), Trstue ('ulture (.ahrrratnry, I)c!":rrr- ment c+f Anitrrmy. University ui Tuas Medical Itranch. (ialvectrrn .4 (' 1 m tn ~R 29 n

( r-n(r-rrrrr r irr Rrtrnrrh nn thr Rnoh-rhrrnl+y if (•unrrr, r:rl•cc S!, (,q, April 196 1 l I I It (' frantee: rtrnrcrat I I'rrrlrn.r ../ .rnrfr: ('ytt,hoFici1 t•.itninalinn (if irrarliatctl cclf. uf whrdc nrl-:rnmns tI/ nr:rny wnrkcrs invite ~tutlies in this hiltl, with Ihc u%c t,f ccll cuttrrres• n( tnccraive Rimma riili.uir.n on the mqxl.rl clirtnm ~trrual numt,cr (if a human :nnnir.n strain. I'rnrrrFrrrr: A hnmin amninn cell strain with a mndal value (if 7S chrmmo- s,nncs wis cultivitcd for tw(, days prior to irradi:rtiun with IINMI r (if r:tmma rays ftnm a r.rdir+active cobalt snurce. lhe (nc%crvatitrn of chrtrmtntrmct was m,Je with the use of phase contrast micrnscr,py. F'indin)rr: 1 he population of human amnion crlls showed scvcrc injury frt,rn the ratliatinn On cultivation, healthy Ionkinp cells were resttrred; ht,wcver, they now had 73 chrnmcxt+met. Repetition of the trcauncnl t,n the new suhlinc rctlut:rd the modal value of chrmmoanmcs It, 71. I urthcr re- duclit,n nl thit valuc did not occur :ttter irradiatinn ft.r tht• Ihirnl ,ntl even ftsr the /t+urlh tirne Othcr human antninn cells wcrt irradritcQ ft+ur ,irntt tlurinF S(1 d:ryi withnut nrakinR tuhcultures l he mt,tl frequcnt chrt.mntnme rnunhcr at the clevcnth suh-culttue (if the ettahlichcJ arhline drt,l.hcd fmm 75 In 71. lhe atnpFinR of the mrnlil value at 71 invitct furthcr ttudret cun- cerninR this type uf radioretistance. "C(1Mf. ( UMrARATIVFi Ff Ff:("ISOf SMOKFf) rArF?R.1(/11A( (Y) ANI) ('I(iARf I If:S UN ('IIROh1OS(1MFS IN l'lIlf(1'• Hy Akin Awa, Yatuahi OItmiki, :(ntl (' M rrrmcr:rt, r:t,,drna (('al ) I r,nnrl:ttitm fnr Mrdicil Rcsc:crch, Irrnt Itryarrrt nrt Ifr,rlnAv.nrJ Alydncrrr, Vrd 19, I,:rFc's tIR-52R, I all 1961 . ( 1 I R(' I t'llirwc Awi :rml Ohnnki) Mitnlic and chrnnrrnnmaa an.rly,,ra wcrc t.rrncrl rrut in tht III (• slrain cclls cxhrctd drrcctly tn Iht' tnwkr collcctcrl Irntn ( A ) whrrlc ciF- :trt',tcs, (I11 t0h:tccn alnnc. anrl (t ) I,.rf,cr .rl„nr Mrnl,hnh-pi:rt trha•rv:r lirms rt'vcalcd that rafxr tmrrkc inthttrd vcvcrc vaturtlv:rtnrn of the cytn- I,lasm tnpNhcr with pycntrtic nuclei in mt,at ct'll. In ctmlr:ttt• little cvi- dcnce (if ccllulit tl:tmafe wis thuwn hy exrtnure of smnke frtnn whtrle cigarette antl trtnn tobacco nlnnc. 'Ihcre wis a decteate in the rate of the mitt,tic index anrl a ctrntid- cr;rhle increiec in the nunrher of ahnttrmnl divisirrnt at a result (,f the trcnl- ment with cach suhdance, rirticularly with pilxr smnke. ('ontraclit,n rtF clntrnu.ann+es was nrtticed in snme metarhaect etpa-,uJ to trntrke frurn whoIc ciFarctlcs and frnm tuhacct, In enntrasl, p:(rcr smnke hrrttlucctl a )•h incidence rd chrmmosnme stickinett. 1 hcse mitotic ohnnrmalitict -and the thifl in the mtwlal chrnnrtr.nmc nrntt,cr rrf thc III C strain are briefly discutacd (lfhrr Frnnlnr: 11 S Public 1lcahh Scrvice. '•1'itI MONARY NFOr1 ASMS rR(111(I('I'F) flY Mf?lI1YI ('t1U1.AN- l IIItINf IN 1111! ' Wltlli? 1'1'KIN I1(1('K " Ilv R 11 Riyd,,n• I)cf.trt. urcnt rrf I',thnhrry, l)nivertity nf l etat Medical Ilrant h, C:rlvcslrrn. ( nnr rr ltr,r,rrrlr, Vrrl 11, I,arcs S71 574, Miy IVr.I /'u.p.r•, r./ lhr rrsl,irnrrny tr,.%ct in the rluck it timil:n in nrany w. ~. t.. iho o tn mnn In prrvn,ua c.I,rtinrcrHS cartinrnn:rs an l aarcrtnt:rt ui I n III, n11ir:,tnrv tr:ut ~,I white I'rkin rlntk. pivcn :tn in1r:t frtchr:rl injection (if mctlrylchnlanthrene sutpended In pnlytorhate RU Mnre detailed invcsliFatinn wat considered desirable. I'rnredurr: Sixly-sit dncks were given 3-melhylchnlanthrenc sutrt'nded in a I (~c' rccnl aqucnut solution of polysnrhate RO, and 2S,were given (mly the pulysnrhntc. A small catheter was placed in the trachea thrnuRh the external I:trynx, and t,ne 25 ntl. injection of the solution was given within an interval of 2 fn 1 minutet: 250 mg. of inethykholanthrtne was given Irr 22 duckt, 125 niR. to 2R, and 62.5 m6, to 16. The birds were sacrificed or died at varying inlervalt, and the respiratory tract removed in tntn for exantinatinn• FYndinpr.: No tumors occurred in birds receiving polysorbate alone, but striated amyloicl in the livcr, and local areas of necrosis in striated muscle, appeared in some of thesc ducks. Many neoplasms occurred in the retpira- tnry tract of birds receiving melhylcholanthrene, the first appearing 56 days afrer injection. Sonu of Ihe tumors were neurnfibromas, and olhers were RnnRlioncurnrnas, hemnnginrnas, squamous cell-Iike carcinnmas, adcnucar- cinrtrnas, antl unclassified types of tumors. Frequently a single tumor showed n variety of histolnRical paltcrrn. No metattases occurred. Methylchnlan- thrcne cryttalt peranted in the tract for long perinds. Since amylt,id and ninscular necrnsit appeared in treated duckt and the ennlrolt, these eRectt do not appear specifically related to melhylehnlanlhrene. l he prohlcnv of amyltridosis in the duck is nnw under study. I)th.r Rrnnlnr: ll. S Publie Ilealth Service. "I.FSIONS IN I)l1C'KS (71V[N MLiTF1YLC110LANT/1RFNE." fly R. FI. RiRdnn, M 1)., and S. M. McAnelly, ff.A., University uf Tcxns Medical Ilranch, Galveston. Archives o(1'athofngy, Vol. 72, pages 4SS-4h4, October 1961. (T.I.R.C. grantee: Rigdon) Prrrlraar of atnrlv: In a previous experiment neoplasms developed in the resprratnry tract of white Pekin ducks following  single inlratrncheal in- jectinn of inethylchnlanthrene. The present study was matk to observe the pathologic changes occurring in the respiralory Iraet preceding the develup• ment of Renplatms. Prnrrvlnrr: Twenty milliliters of a solution of 50 mR. (if mcthylchol- anthrene in polysnrhate was injected by tube into the trachea of 21/ ducks 36 days old: 2 af Ihese birds were killed S'/i hours later, 2 after 7 days, 2 after 11 dayt, antl 3 died on the 26th and 27th day. One was killed nn the 35111 day and Itl were killed on the R4th day following the injcclion. len other hirds 4(1 days old were given 6 to 9 daily injeclitxts and 111 nlhcre were Riven 10 mR. ftir 7 to I(6 timet; all duekt given multiple injeclittns either died or were killed al intervals of 9 to 72 days following the firal injection of Ihe carcinogen. lltt• respiratory tract including only the lower Ihitd of the trachea wat removed ond several seelinns taken ftN hittolt+Fic tUrrly Fin.NnR.: An acule and chrrntic in/lammalory reaetinn (n-curt in thc lunp. of whitc rekin ttuckt ftdlt,winR the inlratracheal In+eclirm n/ tnrrhvlth-d nnthrt•nt In rnlytt,rfr:rtc RI) Atatxinlcd with fhit inflanntntnry rcat Inrn i. rcrcncratinn, hyFK•rldati:t. nnd rnetaldatia nf fhc hrtxtchial chitht•hutn Mcto rlisia nc'curt at early at 7 tlayt after treatment Ilcmanfitmrat. adcm.r:tr cint+mas, and filbrtr.atcr+mas arc rrescnt in the IunRs 72 dayc h,lhrwinp. the • m ~„ ll tn m r

first ttf 16 intralrachea) injcctit.ns ttf the carcinnFen. In view nf the presence of aquamttus mctaplasia ant) tihe ahaence nf typical syuamrtus-cc1l c:trcinnmas in the IunRs of Ihcse ducks the yueatinn is raiaed refcrahlc tt+ thc fiithrtRencsis of thccc neoplasrns. Melhylcholanthrene crystals mechanically rthalrucl thc Iumcn of Ihc srnall bronchi resulting in atelectasis and suhscyucntly Pul- rncmary fihrtnis with an assnciated pulmonary eniphysenia. (3thrr Rrrinrr.r: U. S. Public 1(ealth Service. 11. llumnn Lung Studies "ACUTE EFFECTS OF INNALATION OF CIGARf:71 F SMOKr ON AIRWAY ( ONI)U(-TAN('E." fly )ar A. Nadel and lulius II ('nmrt.e 1r , Cardinvascular Research Inslitute, University of ('alifnrnia Mcdical ('entrr, San Franciacn lnurnal eJ.(pplird !'hy.riafrrry, Vol. 16, f+aRcs 713 716, luly 1961. (7.1 R C. grantee: Comrne) Purporr n/ .rn.r( r: Previous studies of the acute e0ects of inhalation (if cigarette smoke on "airway resistance" are of doubtful siRnificance, a) because sonic of the methods uset) require an initial deep insPiralion, which we nnw know may itself decrease airways resistance, and h) hecause none of Ihe tests mcasured airway resistance alone hut rather a combination of faclnrs, includinR tissue resistance and voluntary muscular efltut Accnrd- ingly, we used the body pICthysmnRraph techniyue to obtain a sensilive measuremcnt of airway resistance ahone. Prnrrdurr: Farh suhjecl sat in the elnsed 900 liter btwly Plcthyamngraph fnr a few minules, then panted thrnuRh a flownuter for several eecnndc, during which time his airway resiatance was measured at flow ralcs nf 0 t liter per second. We then closed a shutter hetweFn the mnulhpicce and the flnwmeter and measured the subjecl's thoracic Ras vnlumc (7(iV) pIcthyc- mnRraphically. F.ach test consisted of four ennaccutive meaawemcnls uf airway resislance and 1GV. We calculated all data as airway cnnductance ( I/resistance )/1(iV to correct for variatinns in lunq vnlume durinR testinR. We tested 4R suhjects whose IunRs were mtrmal, including male am1 female smnkcrs and m.n-smokers, and 22 Patients with a diiRnosis of rulmonary emphysema, aalhma, pulmonary fihrosis, sarcoidtxis, or mitral stcnosis, all eiRarette smokers. Fin.FInR.: In the healthy suhjects there was nto siFnificant tlitlcrence he- twcen male and female smokers and male non smukers, hut airway enn tluctance/f (iV was siRnificanlly hiFher in fernale ntrn-amukrrs in tltr cunlrul statc We sttnlird the acute eflecl of cigirettr smrrkc inhal:ttitm in 16 hcallhy suhjctta 121 anwkers and IS mm-smukcral and 22 snutkcri wilh carJirr- 1+uhnun;trv tlrar7se; the airway ettmhrctance/I(iV ratin Actrc.ractl aftcr thr rnh.tl.tlirm trf ciRarette smrrke in hrrth /rtw/rs I he rllrt ta wrrc ahnrt.t im nrr•r1r.tlr; Ihc nrrart thnatirrn of cllt•tt wna lt rninnlca Irnlhnlalirtn trf 11 S t.rn~nt r...lr.~rrrrnrrl :rrr.,.rd rrvrrtrr) rrr tirrvrnlrrl IFhr rltrtt rd tit•:rrr/It' I thc ,1., rr- r,.nJur I.uu r 1 t i V t.rn.rJ I,y rnh.rl:tlrnn . i rrrrl rrrh.tL,Jr,-n ,•1 na nlrnf Or f Our stwly indicites that mild brnnchoconstrictinn, insuflicirnl trr cauae symptorns, rrcurs immediately in most individuals after inlctliliun trf ciR• arette smnke, lasts 111-R(/ minules, anti reoccurs with a second cigarette We suRpesl that thert are insufficient and inadc9uate data at present ttt warrant relating the "hronchttconstricticwi' of smoking causally to the initiation trr afRravaticm of chronic pulmonary disease. ()rhrr Rrnnror: 11. S. Public 1leallh Service. "Ttlr PA711OL(X:I(' ANATOMY OF THE BRONCIIIAI- TRI_-.f; AND LUNGS." Ry Stanley P. Reimann, M.f)., 1)irectnr Emeritus, Inatilute for Cancer Research, Philadelphia, Pa., and Member. Scientific Advisory Iloard to the Tobacco Industry Research Committee, Archivrs of 1'nrhnfnRy. Vol. 70, paRes 757-762, December 1960. (T.I.R.C. grantee: Pathologic - Anatomic Study Group) Twelve e><perienced patholoRists from various parts of the l/nitcd States underlnak a study of the pathnloFic anatomy of the bronchial tree and IunRs in spccinrens ohtaincd frurn routine autopsy serviees and surRical e~tcisions. 7 he object was to cataloRue chan6es, sites and ineidence in theee orRans in relation to aRe, se><, uccupation, place of residence and work, use of tohacco, and canse of death 'I he classrficatitxn used were: normal, hyperPlasia, syuamous metaplasia, atypical metaplasia, carcinoma in s/rte, careimoma. About 3,(NK) cases were investigated yielding R,590 slides from rnales and 3,661 slides from females adjudged suitable. The border line cascs prn- vnked disagrcemenls, many of which could not he reconcikd. In addltinn to cnnferences, selccted slides were eirculated and many pathtdtrFisrs re- examined their own slides a year or so after their original diaRmrsis Sonic of the limitatiuns e.f the practice of palholoRie anatomy were discuaaed. 1'rom raw compilations the results were equivocal. Rigid statistical appraisals were made and were published in seParate parera (F It. Wilsttn & M. II. Ourke, 1957-60). It was shown that a eot>(+cralrve s(ndy is ncctlcd in problems in which large amounts of data must be collected anJ in which basic prncedtucs in a particular discipline can be collectively asccaacd. "A SfIInY O1' TIIE TRM'IIf:O1lRON('HI/41. fiPl7lll?I IUM ANt) CIIANGf:S RFI.A I1 1) TU SMOKIN(3." Ily K. R. ('nns, M I) , IAmald V. Wals, M 1) , G Keith Palmer, MO., and E. D. Warner, M11 1), 1)t-part- ment of Pathuhtyy, Velrrans Adminislratinn Ilospilal, Itrwa ('ity, I:r lrnnnnl of rhr l.nrn.Crnrr M1frdi,nf S'ucirty, Vol. 51, pages 117-14(1, Match I'rht (T.I R.C. grantee- Warner) rnrpo.p of arndv: Of the first 915 autupsies at Ihe IlnsPit:J durinR a sitt-year LeritKl hrnncht,Renie carcinoma was diagnosed in I lV caacs ( I S fx•rrenl ). '1 Fhe purlNrse of this paper is tro discover the dcfrce and tvhc trl met:rltl.atic chanPrs rtcc•urrinR in the bronchial epithclium :utd It, delrrnnnc whcl'ter or nrrt thesc changes are related (1) to previnus smtrAinR, :rntl (2) In the dcvehq+menl rrf faonchnRenic eareinnma 1'r... rrhrrr r I rflren rcl+resrntalive scctinns nf the trachcn and lhr larpr anrl .mall hrnnthi wcrr arlrctrd in each of 1411 nutrrpsy caars rrrrvrd .urLrblc ha atudy, intluthnr. 26 paticnts with carcinnrna of Ilrc luny lhc ar t lnrna wcrc clasai/ictl aa m,rrnil, hyln•rplaaa, afuirnrruc rnct:t/d:r.ra- al~lrrcal tnct:thlasia, carcinnmi rn trhr, and c:vcnttrm:r I hrre wcre 11 nnn amrrAt•r., i 11

I'/ IiFhl, 11 nnxlrrate, SG hcavy, and 21 very hcavy crn„kcrc am„nR thc 140 ca~cs. Findinrr: Thc hr,+nchial tissucs of only 49 percent nf the cavca cnnhl he c„ncidcred nr,rn,al, cnmpricinR 44 percent of the ct•ctinnt frnm cm„Acrc and 61 hcrccnl yd Ihc secti„ns frum nt+n smr,kcrc ( arcinuma wac f„und r+nly am„nF cmr•kerc, ac was carcinoma in vlu, and the tatter wac fnun,l ,,nly in r-lticnts wilh IunR canccr: the 7 indanccs fnund were fcwcr th:m ,mc wr+uld ct(+cct if carcim,n+n irn rirre had preccded the devctuprncnt ,d carcim,ma of thc IunF Attht.uph the sludy fails to support thc prcmicc that carcin,mta ariccs frr•rn prc ewicting atypical mclaplaslic chanpcs in hrnnchinl cf,hhclium, r snrnkinR level of FfA. There was essentially no change in the cCrum tt,tal chnlcctcrnl or IriRlyceriJes during the experimental peritnl. I hcrc was a sliRht increase in I FA over a 20-to-40 minute period in the nnnsmr+king contrnls sliRhtly afa+vc that anticipated from fastinR alone. l he five suhjects who "chain-smokctl" all developcd FFA elevatinns which penisted duriuR the 60 minutes of ohservation. An eventual, delayed increase in serum kvcls of cholesterol and other lipoprotein lipids, as occurs in animals after epinephrine, is a conceivable result in humant after smnkins. This, at kast, suRRests an e><plartalinn 1(or the higher chnlesterol levels often found in smokers. If, as many invecti- il ic evident that thcrc is a ciRnifrcant acsnciatinn between certain cpilhcli:rl )Catnrs believe, a disturbance of lipid metabolism is a factor in the dcvelr,p- ch:,nFcc in the tncheohronchial tree and the prcccncc of m;rlil•nancy. It ment nf athcrosclerosis, the effect of cigarette smoking on lipid mctaM+lism wnuld accm that a histnry of cigarette smukine is ciFnificanlly rel:r,c,l to the sM>,dd be given attention in considering the palhosencsis of this dicnfder incidcncc r f carcinnma, and Ihc rcfnre difficult to iy-nnre A linal ,Iclcrmina- \ Other Rrentor; (1. $. Public ffeahh Service. ti„n r,( thc rcl:+linnchip between trnokinR and Ihccc relh„h,pic:,l clcmpcc will "T'IIf: REI•AT IONSI(IP HETWEEN BIOCIIf;MICAI- A(TIVfI Y ANI) rc,lnire rnu(h mrar shrdy. MEC11AN1('A1. FtINCIION OF T7ff; I•IEAR'r." fly R. J. IIinR, M,1). 1I1. (:rzrrlitrrn.rrrlrrr Cflldir't "IiI 1 fY'r OF ('ICARETTF. SM(tiKIN(i ANf) NI( nl INF ON SI?RI1M I It1 1: . I A 1 1 Y A('II)S." fly Alfred Kerchhaum, h1 a) , S:,nmcl Itellcl, k1 I) , I-r/w:vd R 1)ickslein, M h., and I ennard 1. VcinhcrR. 1'Fh I) , 1)ivisinn nf ('artlird„Fy, f'hiti,Iclphin (icm•ral Flr+cpital ('irrulnrirrn Hrtrllrr lr, Vol o, p:,lcc h11-b1R, May 1961. (T.1 R('. grantee: Iscllctl Prrrpnrr rrf .lnrfr_ : It cecmctf important to invcaliFatc how the cRecls of nicnlinc inlloencc thc metahc+licm td Iiritlc, rarticularly since rcccnl ctudicc h:,ve chnwn that free falty aei,ls (1 FA ) are rapidly relcace•,I into thr circn- lali,m frnm tiscuc fat dcprxitc f,JlnwinR the athniniwitir+n ,rf cf,incrihrinc and m+rcf+incl,hrinc and following s,ress cRccts on !hc cympalhclic ncrvnuc systcm 1'.nr~dnr~.: f'aticn,s and pcrsnnnel at Philadelphia General Ilnspital were sclected at random, includinR 7 nnrmals, 7 coronary heart dieeace caeet, and 17 palientc with varivut other tliaRnnses, aped 16 Io 72• including m+n-smrrkers as well at smnkert All lests were m:rJc at Y a m. in f:rslinF condition In 17 tnhjrcls, 2 eiFarettes were smukcd within a I1/ minute rc•rirH1 fllraal saml+lcs were taken hcfnre smnkinR, immetfiatcly therea(ter, and at 111, 20 and 41/ minutes after ecssatinn nf snu+kinF- Another prnup nf V suhiccts cerved as cnnlruls to dNermine what elTect the rf+erirnent:rl pnKe,lurc itcclf wmrld have f fastinle, prcraratinn, venif,uneturc. rtc 1 Scv- er.I r,f the crrnrrr+ls inhaled deeply to cimulale smnkinr rlurin(• thc initial tll minrric fK•rir,d liach n/ S suhjccts annked 6 cif•arc'tlcs in 411 minutcs tn stnJy thc rllrct nf "ch:rin tmnkinP" am11 VA enncentrilinnc wcrr rlrlrr,ninrd at 10 minutc inlrrvils for a Intal fxri„d ,if hll minutrs :+flct cm,,krnR nrJr•d Fin./inje.: In :,I) 17 tuhicctt whn srn„At•d 2 cit•arctlrt Ihc I I A Irvcl% rr,.c at Ihr cn,l „1 Ihc .n,,,kinr LcrinJ, mnsinr,rm elrv:,ti„n ,KCUr10 numrtr. ler,r I wcnty ymt •1t1 rnirmlrt nllrr trn„Ainr Ihr I I A Irvr1% werr .,,11 :,h„vr th, I„r.rnnAinl• I, v, I in mr,.l in.l:,ncrc I lie I 1A rry,nn.c ,h,l rn,l :y~l~c.rr h, I.r rc/.rrrJ 1„ c,/•,•, tct. tnr„kinl• h:,l,ilt, tvl,c r,f (il•:,rc,tr u.crl, or I,re- t ( 1'rrM'r~dln~r, Mh Inrernarlonaf Conerett o/ Internal Medir inr, llasel, Switzerland, 1960. • RRecent studies have further defined the myocardial utilization of fatty acids, and reveal that, in the fasting human, the mean myncardial extraction of free fatty acids accounts for 42 percent of the total fatty acid ealraclitm, while the esterified fraction makes u the other SRpt rcent. Since r+leic acid appears to be the fatly acid primariPy used by the heart, it appears Ihal, in cnntrast to previous conclusions, the myoeardium uses preferentially un- saturated fatly acids. In the most commnn types of heart failure, ahsence of alterationc in ulilization of oxygen and suhstrate by 1he heart probably denotes absence of invnlvement of metabolic factors within the heart. llris by uclusir+n points In the organs of energy utilization. the contractile proteins, as the site of the derangement in the myocardium in this group. Sludks undertaken nn actnmynain hands, although not yet conclusive, support Ihis cnMentinn. Thus, it has been sflown that the contractility of these bands from hearls nf human suhjects dying from congestive failure is impaired. While there is nu doubt that the blood concentration of catechnlamines influences myocardial funclinn, the pharmacologic role of catechr+(amines stored in the heart muscle remains unsolved. The release of stored catechnl• amines is nf inleresl fxcause it is rekvant to the main topic of (his dicfus- sion, the relation af funcliona( activity of the heart to the integrated enzyrne activity. The pharmacoloRic impnrtance of the stored calechol:rruines is as yet uncertain. 1 his review auR`ests that work on cardiac metabnlism has nrnved frt+m the scarch (r,r utilizntinn of suhslrales by the heart Io the invcstiFatinn n/ the re(:uir+nshir hetwrrn biochemical and mechanical funetinn "MY(X'AItUlA1. ItI Sl'(1NS1? TU ('I(TARI1-Ifi SM(1KIN(7 IN Nlt/(. MA(. Slllt)ff('1S nNl) I'A111:.N'IS W1111 CURONnRY 1)t\I AS1 " Hq T'. 1 Reran, M 1> M. I f rank. M 1) , 1 /~. Mc(~in1y, M 1) , 1 7rJrl, K~1 1) . II K Ilrllcros, fN 1) nn,l R 1. 11inR• M I/., 1)trirtmcn/ r,f Mr,hunr, Wayne State (Inivetcily ('<dlrFc nf Medicine, f)etrnit, Mich. ( mrrLur.•n, Vulume 21, paRet 1G5.1h9, M:rrch 1961 (1 I R('. Frantre 11inR1 m tn 1S „ 14 R~