Tobacco Documents Online

42 ~ TR HN 0 11577

I . I I I I r m m 1 THE COUNCII, FOR TOQACCO RFSEARCII - U.9.A. r

~. : Organizatipn and Policy r ~ TAe CoundJ for Tobacoo R.eseuth-U.S.A. ia the sponsoring agency df protpram ot re.eares Into questSoos of tobacoo use and health. It was :a', organbxd e" (a 1954 by repres~eetatlTa of tobacco maoufacturcn, I;!•' vvwers and warcbotreeea. Rres..rck h.npported through a program of r '.' A 1ci.etl6c Ad.boq dovd to The Couocll adminiaten this grantF b-aW pc>tr.m. The Board cvrteotty consisu of eight ncientl.ta and doctors 1•So .ralotaln tbcir respect).e Inathutbnal amdatlo.n. A list of the Bovd memben appean oa a.ub.equent page. ~ The Scientific Ad.isory Board has full n`.ponslbiihy for rrse.rrb po1ky and prtgnmminf. As a Board It dod not directly engage in research for The Counci), nor does The Council Ft.elf operate any research facility. dnntsin-ald for research are m.de by the Board to Independent ocl- eati.b who are assured ccxnpkte scientific freedom in conducting their ft.earch. Urantoes alone are responsible for reporting or publishing their Mdinp In the accepted scientific m.nner-throuRh medical and scientific /ootnab aad societies. or.nts by the Board through 1965 have been mack to 280 scientists In d t 100 h l i' Th T i i d h awar s mtrtut,cxu. se 6ran a e wsprt , u.n rexarc era r..% o+rer Te*s t , an 4 .i' . . e..e.trA t! 7M (Wl ' . . i,.,,,, - .... - .- ..... F..,,- ...jrF,,,..,.. .,y .- ............ .. ...~..,..~.. ... -rV Ihi. Report. Also Included are abstracts of 64 research paptn, acknowicdR- InR Council .upport, that ha.e ppeared In scientific journals since the ~ 1964-65 Report .nd through [)ecember 1965. A total of 465 such pqxro ~ have been published by Stant recipients. T1M(YT1fT V. ItARTTftt7T W. T. HOTT (balrvn.e F:arcullvr 1)Im lor T11E Cl)IIN(:11. FOR TORAC(:O RF.9F.AR(:11-1/.9.A. 633 Tbird Avrnu., New York, N. Y. 1()n1T i y I ANNIIAI, RI':1'(1RT of the SCIENTIFIC I11RF,(:T(1R CLARF.NCF. C(/(IK I.ITTIE, S...1).

SctENTtFtc AtrVtS(1tiY tlOArlt/ to 1 hc ('nuncil for 1 nhacco Rccc:rrch U S A KI NNF 111 Ml RRII L LYNCII, M I) , Sc 1) , I I. I) ,( hnnmarn ( hunr rl4+r anA I'rofrtior Emrrrlut u( !'othnluRv Medical College of South Carolina, ('harlcztrm, Stiuth ('ar~rlina '1'Altl.N: oF <:I /N'I'f:N'I'ti e HOWnRh fl ANpFRVnN1', S(' D. ScirntilSr Frfnor, The Journal of the National ('ancer Inaotutc NethesJa, M aryland I RI(IIARI) 1 IIIN(7, M h I f'ro/rrtnr ond ('hoirmon, Ikl+artmcnt nf MrJr(inc Wayne Statc llniversity ('ollcRc of MeJrcirK, I)ctroit, Mi(hrgan I I I InlnKluctirrn S hc Prtrrrctt anJ 1'1;rmtinl; ttf Rccc:uch K le.h.v;rctt nf Rcl><,rt. 26 M< KFFN CA 1"i hLL, Pn D, M 1) ~ I'rn/rr)rv Fmrrilut oJ Phormot nlnRy P.ychu Phyiitdngical SluJic% . 27 ('orrxll llniversity MeJical ('rdlrRc, New Ynrk, New York I ~ ( arJitivatcuLu Studies . 29 l.hON 0 IA('OI1SON, M 1) I)ran of thr I)rvivnn of Rrnlnqn of Sr rrnr rr ('arcinrrgc•ncvia SluJica 40 RrRrnrrrin 1'rn/rnor nf irl 1, rrn, rr I l lnivcraity nf ( hiciRrr Studic. ;rt thc ('cllular I cvcl . 41 !)i.rrrrn, ArRirnne ( antt'r RctrmiIr Itivtpii.il ('hicaRo, Illinoit I IriJt miuhrry . . . 44 ('I.ARFN( 1((x)K I 11 1 I I. G I) , I 1 I) . I rr t U I'ulnurn;rry Phy-,iuhrRy . 46 .Grrnrrfir Ihrrrtar, Ihc ( rruncit hn Ir~het( n Kc•t•.nch I SA f)irr((nr Fnvrur.r, Ruu'trc I( )acksun Mt•nurnal I aMuatury I ( hcmi.Lry nnJ Ninchcmi.try . 49 lfar Flartx,r, Maine I i SrANLFY P RFIMANN, M I) , Sc 1) ' Pharrnacuh,Ry anJ P.ychn t'IcrrmactrhoKy SI anccr Reccarch f)irrr m. Frnrrifut, 1 hc Inalitutc /ur ( OIltt•r StuJit'% . S7 Philadclphia, Pennsylvania I WII LIAM F RIFNFIOFF, la , M I) I I'rn/rfror Fmrrirur of SurRrrv )nhnt Hnpkint l)niversity SchtKd rif lvlcJitinc, ttaltirntrrc, K1aryl;mJ Rct'ilticnlc nf Gr;rnl% SV R(Hlhf(1 (' II(K Kf 1 1, f'rt I) I MrtHRISUN I11(AI)Y, M I) A rroa iolr S rrnrr/rr I)r.r( nrr f t r,-, mnr .Ir vnrr/r, I)nrr hor r

Int.roduction l 7 ...3. .f.. t 1 L.~ ~ <r, d ~ a, t1 ~ In the 19(,4-hS Report it was mentioned that over the yCars er<reri- + mental and clinical evidence to support the thesis that cigarette smoke ercr- ciaes a direct carcinosenic cffect on man has not been forthcrrminR After reviewing another year of research this slatement still eppresaec the state f of uar tcicntific knnwlcdRek fivcn fat•d'a-1ed or cprecled epidc•mittktFicr+l evidence whic'h the ciga rctle carcint+Fcncsis hypothesis rcyuires, such ac a narrowing trt thc gap In•twccn dr.Hh ralca from (unR cancer in the sexcc, owing to the Rrc:rt increaac in amtrkinR among womcn, has failcd to maleriali?c. Indead of appearing at increasingly earlier aRca fnlhrwinR an increacc in smoking among younger people, the peak age of incidence t+f lung cancer shaws, if anythinR, a shifl toward oldcr age Rroupc in recent dc•cadca I In addilittn, a numher of studics throw doubt ulxm file accuracy or diagnosis of primary cancer of the lung in a siRnilicant fx•rccnt:rRc of caaea• ac refittrtcd on death ccrtificales. Thccc ctudics ahuw that aa high ac IR-211'S, nf rcctxded c:nKera of the lung are actually metaataaes rrf cancer from t»hcr txr:rna •'1 hcse eludics highlight the need for widespread and concerted effort to cstahlieh more accurate and more uniform methods nf tccording thc incidence of lung cancer. Recording of this sort chtwld he a haaic Iwhlic he:rhh recla,naihility Similar prr+hkmc with respect to accuracy trf diaRmraia are encnumercd in the study uf the other constitutional di.c:nec which have been rcptxlcd to he associated staliatically with eifarclte Sm<tkinR Such Lrnhlcme are marked in file caec nf file chronic reapiratnry diacaas where lirm clinical atandards and dea'riplinn have slill nn/ Ix•cn dcvchqx•d It ia obvious that reliable cpidcmicrhrRy mual fx• Irrcmiacd tm rcliahlc diacacc incidcncc and mortality data. Ac mtne and more medical men and I:Jwtraltrry rca•:trih wtnkcra rcc riFnirc :tnd Lcctrnrc familiar with the ctrmplcnity c.f hurnan Fcnctiia. o muth mtrre reatiatie :rttilude k LcinR dcvc6qa•d hrwird file nct•J of huther anJ nurre rctincd atudiea in this ficld lht•rc ia inirc:rainR rccr+Rnr/itm Ihal in thc dt•vt•Inprnt•nl rr/ tmn.trru tirtnal dr.r:raca, auch aa IunR eanccr, inJivitlu.rl Kcnctri tli/firentt a.ric irulaut:urt I hc iJr•a th.rl thc pcnctii ha•ia rrf di.ra.c t riltin nru.r I•c .1 vml4h• Mt n dch:rn I ht nunrrmm Icra Itmr, ht•t•n nhatrlctr, hul Ihc :rb.urJ. 4imrnr id thi, S

unat,hhiatrcalcd tt,nccpl nl Rcnctics h.ra IK•rn Painfully tlt,w antl t,F.alrutlivc It, a rmxc rcalialic undcralinJinR td Iht• cnml+lct n:tturc trf Ihc yrnrtic crtilc and prt.ccs. I ach t,rr:,n tyttcm and tiacuc r,f any inrliviJual h:rt itt t,wn '•Lvill in•' level ttf atucnFth anJ of wcakncss, t+( autccrluhihty t.r t,f rrai.t:,ncc u, thc rhallcnrct t+r the cnvirnnmcnt, tM,lh from cxtcrnal fatttns anJ frrtrn imrrrctl unh:,lantct r,( function. (•t,mpirativcly few people •re sn cnnstilutt•d th:rt :, ttm,l'lcrc intcrnal halancc hclwrrn diflcrcnt r,rRan aytlrmt and lia.uca ia m.tinl:,inrtl Grr nine t.r Icn tlct'atlcs r,f aginR tt, that they c't,mt• In Ihc rnrl nf Ihrir Irlc ":,II at nntc" and at a fhal:mcrd unil S.,me ty.trrna, Irkc Ihe rrltrtKltativc, arr Iim,IcA hy Ihrir inhrrrnl n:rtutc Irt a thrntrr Iw ritr.l t+l activity than art• tHhcrs Hut trll aytlcma cvrmually .httw the cti(-ttc t+/ the wcar and Icar t.f lifc And il it Ihc t,rdcr in which. anJ t•.Irnl tr, whith, thc v;,rittua tnRan ay.tcntt and liaaut•a aKc atttl aht,w aip,n rd dnnin iahinr, irrcRnlaf tv ahnr,rmal luntut,n, that reflecla the Rcnetic pallern with whith the intlividuil it cntk,wrd Vcry cvitlcnlly thctc cai.tt a vaat numl.cr t.f, ia yct, htllr un.lrr.trwxl tyht•s and drrreea rd imenel.,tirmalul e Iytwren r,rFln .yatcma and t.etwccn thcir ct+nrl-ncnt liaaur. Ihcw tornldrt.rlt- iny tlter :rnJ "au:u/•ht Iinc" rclitiunahrf, IK Iwr cn Ilhc /tm. Il,.rl nntlt rht' nrr:tn ttr (rtCUC I4ttl'lltJallll(•4 and Ihcir fni,Prt.avc JcvcLy,nn nt ,nJ .nlln.tmtnt rn tfu• tt~mfdt. :mil th:,n~ inR ct,amtn ur Ihc ,ttthvitlr,:,l't Iwuly Ilut Ihc hral .Icl• townrtl Iht• cvcnlual an:dycic anrl wiac utiWititm nf rhr •rtuanra, n lu rcttq'.ntic 111.11 it t•ai.la the in,linl phaset t,f the t,rRaniralitm of mtnc cttcn.ive anJ rntcnaivc rcac~rth in thia hclJ arc umkrw:,y fhy I hc ('t,uncd In I,l:,nninl! Ihn ftrtrFr:,rn. consultation with thncc nr,w active in this type nf rescarch h:n hcc n cstah- lithcd and will t.c continued I he rcrsiatcnt slrikinR e.ccts r,/ IunF c:mccr in m:rlrt inJie,tcs II,:,t (nrlhcr knowlcdRr of cnJt,c'rinc cttnthtit,ns which accr,mpany Ihal diccaac is tkairahk Not nnly thc Ronida hul Ihc pituit:ny, :ulrcnnla antl IKrhal,a rtthcr cndt- crinc p.Iantla may, if rnnrc carcfully c.:,mincJ :,t aulrq,ty antl tltning thc ItrtKt•+t rd aFcinF. Rivc hiattq+atht,LoPical cluct lu hcll, in Ihr an:,lyaa t,f the t,h.crvcd ua dillcrcnce in lung canccr int'itlcncc I hc tlutly r,f r•nth.crinc lvnclit,n :,fqdica nrtt nnly It, the tlwly of Irmy cantcr F.ut tt, tNhrr Ir.rma uf tantt'r antl tt, certthnva•crrl.u rliu.ra a:,. wt 11 N1rnh rrnnc infnrm:rtittn ia nrct)ttl t,n II,t• trtrnlr:,r:,livt• h,.l rlr:rlhutnyy ttf v:,ruruc nrR.,n v.tcmt and Ua.uct whrtl, n,.,y Iw th.vattt•ri.tit ur Iht•.c tlr. r.ra a m Iwrth .. tr. antl •t drllt rt•nl .rt•r. t In auc•h alutlics we arc again faced wilh painslaking, lirnc cr,nwminR retcarch, hut the sountlncss nf cflorts tt+ ohlain as ncar ly a cunydclc /,icttnr uf dtc intcrnal hnrmnnal funcliun as is I+r,ssihic cannut fx yucstioncJ '1 he infr,rmatit,n Ihus ohlaincd will incrca.c uur recaFnitiun r,t Ihc cnmplettity r,f Ihcse tliscascc, make clear thc J:mRcr uf ovcrcml,hasis t,n any single fac•Inr in their etiology anJyhus dcvclt,p a rational and cr,n.trutUvc st•nse of scienlilic and lay persrectivc ca+nccrninR their utrgin, dcvchq,ment anJ evenurally their prntfa,ncmcnt or prevention. / hc past year has been charactcr'vcd by an especially inlensive cflorl tn analy>'c langress during Ihc (ast thirteen years of I hc (•ouncil's f,rr,gram ('rilit•al gal,s in knt,wlrJRe slill rcmain which nccdctl ttr l,e t'Icarly Jchncd, anJ cr,ntcntraICJ hlanninR has been umkrlakcn tn tlcvclt,p mcthtwl. wluth may fill w.h Raps and prnvitk murc dkcisivc au.wers to Ix•rarslcni kcy queslions. 7 h

'I'lic I'rc~bress and . Malltllllg of ~~CSC~Ii'(l~l "The Council has consistently placcd cmphasis urnn direct studies of the respiratory systcm tisstxs, both in animal rrx~kls and in human patients Sincr ncnplastic druaxs of these tisnucs and partit ularly in t1rc• IunR are of sf+eeial errncern, prtrhkms enncerninR the etiohrRy ol (+ulrrxin.ry carKCrs havc received malor attention Inlurl~ctt(rn Stu~li(~s Ulnervatinrn of the consequences of exfrrsinR the lunRs rrf eafx•rinuntal animals tn whrrlc frcth ciRucttc smr+t.r have tn•cn a significant Part of lhc ('auncil's f~rc~gram aincc its carlrcct t+cRinninRs Such stuJits art Ntill being cnntinucd, cnlarRrrl and rrhnrrl Icn ycars of I~t•rsi.tcnt wr+rk by several independent ( uuncil clarnaurcd rnvr.trR;dra. and rnany rrthcrs, h:r.•c srr f:rr failed to dcmr+n~tratc the tkvchrhmr nt r+f clridcrnuud I uln>,rnary tumnrs in a variety of animal species following .uttr dnett etlwrwre trr ciRarene smuke by thc inhalatirnr route. Smrrk~ Conden.atYa in tlr~ Lung 1 hcse whole smoke inhalation cxrerimcnts have been snrplemented by nthers of a nMre e><treme and kst "phylioh,gical" nature in which rrrn• Acnrnrcr nf cigarette srsx>ke were inlrrxluccd inlratracheally into the IunRs of scvcral sf+ccics in various kinds r>f sotutions, su.f.cnsiom or cmulsions, both by ('ouncil sf.rmsored investiRatnrs and by rHhers While thcse exl.eri- ments do not rescmhle any situatiun normally cncrruntered by human~, they sre of scicnlilic interest in that they have not Irrrwluced cancera of the IunR, nnr have lung cancers been produced by snwrke crrndensates dehvered to either intact nr damaged IunRs by the inlravenrrua rrrute On the nthcr hand, carcinrryrrnic ~rlynutlr.rr hydrrrcartwvrs such a mcthvlchrrlanrhrcnc and hcnypyrcnc have I+rrrJuccJ canccr% when intrrrluccd into thc Itunl-. rrf Ir.rnnlcrs nr rlut'ks in simil:rr srdutirrrta rrt emul.ron~, and alcn ..hcn nrt~ttrrl urlritrnrru.ly into r:rtc wrth infarctcd IunKs I I I J Cnrn/te ('.rrr.lrruntrt ore nnt SmoltP 11 mn.t l.c• crnlrhatiirctl that cumlcnsatcc nf ciFarcltc smrrkc arc nrd cquivalcnt to wlurtc fresh tmnke in cithcr phy.ical or chcmical (trrqK rtics ( hcmical inrl f'hy.ic;J altcratirms lakc place. .umc ral+idly, in snurkc Irrl Irrwinp it% furnr:uurn antl Ihcx crrntinuc in the contknsatc altcr cnllcctirrn Nrrt only Jrr such crrndcnsalcs lack all of the Ras hhase inRrcdicnts rrf smr+kc hut alur arm(• nf its nnrmal val+ru constitucntt. Shur1-lived caml+rrncnts r,l hc%h smrrkc may not survivc rnntknsatinn and sturaRc, while thc Idrysical %tale of thc smnkc acrn%nl is nhvinusly quite diflcrcnt from that af a cnn dentatc I hc ncRative results of thcse etpc'rimcnta with smrrkc crrndcnaate uc nevcrthcrca% intcrestinR scit•mirrcany sirxe stmx uf thc cvtmlwrncnlc nf srnokc, includrnR :rny It:rccs nf fvdymKlcar hydrocarl>,ros, arc Frcatly con ccntralcd in Ihc cnntkna~tcs so that the tknaRe uf thcsc is greatly incrcasc•d whcn condcncatcs arc uu•d I)iflr•renr. Rrhrrrn Skin an.I Re.pirntory Trnr•t T'i..ur. The skin is ohvipusly a very diflcrcnl ticaue frrrm thc mucu% crr:ncd ti.cucs of thc• rcapiratory syslcm and would f.c cr<la•c/cd to -hrrw rhllcrcnl rccltirnccs to cnntacl aFcnts. Such diffcrcnccs h:wc rccc//1Iy IMCrI Jrmon aUatt•d, for crtamhlc, by exl.crimcnls shnwinR th:rt thc mucu. mcmhram% nf thc oral cavity in mitc arc much mnrc resistant to Ix•nil+yrcnc Ihan Ihcir skins whr n M,th li..uc% are Irt•atcd unJcr parallel crrnditirrnl wnh idcntic:J tlrraca rrf thic anh.t:mcc ( iycuctlc .nwrkc cunt/cnsah•s h:rvc l;ulctl :rltrrl•cther tn Intnluce canccra in thc oral eavitic% nf the camc Strain nf micc :n any drr.apc cn far attained Mnrcrrvcr, hcalthy intact rtxlcnts have failed to tlcvclrrr Luhnon;uy Iumurc whcn suhjcclt•d Irr tln• inhalalian of Iturc funx•c of mcthyh hnLrnt!ucoc ur IK•nrl.yrcnc, althnuph thc•ac tuhslanccs arc I+rNCnt muu.c .kin c;rr.inrrl-cn. Sut'h dirct•t rrh.crvatirms dcmrrnstratinR dram:Nic ditfcrcntcs in .ut cchtihility Ix Iwccn .kin and recpircNrny Ir:rct tiawcs hr c.ut innl•cnic ;p•rott place acvcrc linrit:uiuns nn the uccfulncss a( akin cilr rirncnt. in thc stuJy of lung cancer ctirrlrryY T'he 11ca.rrrr•m.•nt nJ LnnR T'iaxnr E;x/.rr>,rrr.• trr 4nrr.h.~ I)urinp thrw rrn rrr mrnc yc;na rif /vrai.trnt .turly uf tr~h:utir rnnkr inh:rLrtirm hy arunrel., many inytrrrvcmcntc in tcclmiyuc h.rvc Ircn nr.rJc hy a numltr of invc.tir:rtrna Ncvcrthclc.c. a majrrr rlrllirully hcrclulonr ha, been Ihr' I:rtk r f:rnv qu:mtil.rlivc 0hµ•ttivc mc:nrnr• nf Ihc acrual ctlrnurc 9 R

of lung surfaces to smr•ke, esf+cciatly to the parriculatc phasc, in cithcr ani- mals tit humans 1 his lack has prevented rkvrk+l+mcnt tit ar)cquatc infrx- matinn on dr+se response relationship In thc case of human cpidcmioh+Rical studre•s, dcf.endcncc has generally been placed on data derived from qucstionnairrs, fxrth as to the claily levcl and kind of smoking and as to subjective impressir,na of the cxtenl of inhala- tion of smoke A recent Council-suM+cxted study, not yet Puhlishcd, has shown the very pcwir reliability of such recall data by comparing the results of sevcral succcssive interviews with the same subjects at intervala A sepa- rate ('ouncil spnnscsred study has suMesled a suhatantial role nf psychn- kgical attitu<ks in modifying replies to questinns ahout smoke inhalation Practices I hese fmdinRt further empha.i?e the rneed for an nhicclive hir+chemieal methrxl of estimstrnR smoke e.fxnurr of Ihe lungs ('onfcrcnret on mcthrr- drrkrRy have t.ccn hcld, s dcxrrhrd in earlier Annual Rcfxxtt, and uveral r.(+L.ratnry studics have been set up Gime fanRrcts in mcthrx3rdoRy haa been made in Ihc case tit human suhjccla, who quitc uniformly inhalc anw+kc, if they d+r, frum the oral cavity with admiuurr tit trn tr+h+rty vr+lumc• tit air taken in thrrnrFh the nasal fras- saRes Since h+rnun suhlcctt can rcy-md to afv+kcn inclructinns. the hhxxl levclt of carM+cyhcmrrRh+hrn can hr rrlatrd tit al+c•cific numhers of inhaled or uninhalcd (,uf1a, taken t dclrrmrucd intrrvals I his may acrvc as a nxa- sure of Ihr dcRrrc r+f cnnt.cl of alvcr+lar aurfaccc with Ihc fis Phaac rrf anKtkc If the usual mannrr of human amr+krnR maintaina a rclitrvcly fircJ ratio between particulate matlrr and Ris f+haac cnm/wmenls of the amr+ke until they reach the alvre.li, the e.Prnure of IunR surfices to /+arliculate matter couW also he estimated as a fust apprnximatinn, from blood carhrtayhemo- Rlohin kvcls. It still remains, howcver, to apply this and other objective analytical methods that have been under development to a auhstantial laof.ulatinn rd human smokers to assess the variatinna in actual lung ea(xraure to particu- latcs, as a check aRaintt questinnnairc infnrmation I he cnmparativc IunR er<fxnures among typieal ciRarctrc amr+kcrs on the one hand and the •:mrrkers nf Pifus and ciRars, nn the othcr, still rCmaln to tic exl.hued aa Ihcy may he aRected by the various puffing and inhalatinn ptaclicca of individuuls within thcac Rrnufn With animsls, the I+ruhlcm has t+rr+vcn Ir+ ln- rur+rc ditfiruh tKralltc nf their Orncral f.raclicc r+f inhiling tnu+kc thrr+uph the nacal I+aa.,rfca Irn rtr dr•nla, thccr I+acsapr. liltcr srunc fM+rrn+n r+f thc I+:rrtic-ulate maller a+ Ihat the (ati++ tx,wcrn I+.nt++vletca :rnJ F.rt Id+.r.r cr+mlM+nrntt rd Ihc stnr+ke is moxli r t fied in rassaRc and the ratio as it actually reaches the lung is unknown Mnrersver, tbc ability of rodents to suspend respiration for considerable peri.xls of time is a complicating factor. C'arhn><yhemnglobin levels have been measured in the bloa.d of mice following vari<ws smoke inhalation procedures, and the levels have been shown to indicate substantial contact of lung tissue with Ras-ples-vaf+nr phase ingredients. The presence of nicotine, an ingredient chiefly of the particulate phase, has also been denfonstrated in the lung following such srnnke inhalation 1 hese measurements shdw that stnoke has reached the IunRs in substantial amounts Nevertheless. quantitative estimalir+n uf the particulate phase esposure from earbor<yhcmo6lobin levels in the bkxxl will not he justified unless the ratio between particulate matter and Ras-plus vapor phase as the smoke impinRes on lung surfaces, can he determined by snme objective method. (:on/er.nce on TKhni7rrea for Bxl.oarrre of F,xp.rirrrentnl Anirnale to (':iRnrette Smoke Inlsolation On April 21, 1965. a conference was held to consider the improvement of techniques for ex fxnure of experimenlal animals to cigarette srnnkc in- halation Participants included several recipients of ('ouncil Rrarus who have been involved in cnnduetin6 animal inhalation esreriments Iksides memhers of 'I he Council slaff and Scientific Advisory Hoard, several chem ists from tobacco processing companies were present to provide technical in- formation afx)ut the chemical and physical properties of tohaccn smoke Sev- eral guest scicntists representing the fieWs of engineering, pulmonary physi- ology, pulmonary pathology and other rekvant disciPlinec ako anended: 1'a.riripanr. (other than Roard and sta0) Or. Ilollis (iG liuren, 1)irector, Trudeau Found:uinn, Inc 1)r. Young ('hu, Research PhysinHrRist, Philadcll+hia (:cncral IL+.I+ilal 1)r. Arthur Fursl, Uirector, Institute of ('hemical flirrh.Py. ttniverehy ul San Francisco 1)r Ircnc PGrddrinR, Ik'Partmcnl of SurRcry. Allrrt l inacin ( rrllrpc of Medicinc. Yeshiva llniveraity I)r. I crmar+l (irccnhurR, I)cparuncnt (+f 1'rcvcntivc anJ Fnvirrrnmcntal Medicine, AIlK•rl 1•instcin ( r+IlcFc of Mrrhr rnr, Yeahiva l/nivrraity 11r F Itr.mhurRcr, 1'reaidcnt, llir+ Rcat•arclr lnditutc, Inc 1)r Cccrlic I curhtcntk•rycr, I)cl+iruncnt of ('ytr+.hcmr.try, Swi.a Inali tulc Irrr I xIKtrmrmal ('anccr kc.carch • II 1()

11r IlarnhlN Maclarland,Scmrrr Inucr~lr~~r.t,lla~lcttut I ~lwrratnrict, Inc 1)r hnrdc A Mclvcr, Atttnratc Prrdcttur tit 1'ethrtluRy, Mrtlhcal (•rt1 kRc of Sr)Uth ( irohna 1)r, Martin S PrtrVCl, ( hicf, 1)cl.artmcnt td Oral I'athttlrrRy, Ncwark (•ity llrrtpital Ur. Robert 1 Ryan. Department of Surgery, T ulanc l tnivcrttty Schrxtl of Medicine T)r C f larold Stefrce, PathoM6itt, C-af.c f car Vallcy ITtt%rital T)r. flernard Teah, Asuxiatc Oircctot, I rthund I.afxxattuy. University of Noire T)ame Mr Richard Wallon, ('onsullinR FnRincer, Ilmton Dr. Russctl W. Welkr, T)ircctor nf LaTxrratnrict, Menntrral Iltnl+ital ttf ('hctter (•ounty !)r. William W Nates, Jr , T)irector of Rcscarch, I.iRReu & Mycrs 7 ohacco Company Mr Robert (-arpenter, ('oordinator fnr Biological Research, Philip Morris, Inc. I)r. Ti S llarlow, Astittant ManaRinR f)ircclor-Rescarch and Ikvclopnxnt, 1 he Amerr(an I uhaccn (•tanpany 1)r lamcs 1) Mold, Atti%tinl Ihrcclrrr tit Retcarch. I itrRctt & Mycrt I nb.ccn Cnmltiny I)r II II I'.rmclc, Vrtt. 1'rt'r-L nt 1' I r,rrlLuJ l nntLanV. Inc I)r Murr.ty Scnkut, Itucttru r,t kr.r.u, Ir Il I Ittyn.4J. Ttrhactn ('nmltany 1 rnnt thc drtcutwunt, ccrt.trn I+r rnt rpit ~ tit t.tmc tlc:tr I Mechanically generated ciRarette srrntke mutt lie hrtxlucerl under conditinns clntcly rctcmblinR thutc in human srnttkinR if it is to apprnttimate tht phytical and chentical prttqxrtict of the trrxrke as it is in halcd hy humant lhis teyuirct intcrmittcnt huflinR at tuit:thlc inlcrvalt with rcRulation of puR volunx, duratinn, frcyucnt y, and lcngth of hult left unburned 2 CiRarrtle smoke, aftrr Reneratiun, chanFet rapidly with tinu in cer- tain prnf.erties, ntttohly p:vUClc ti?e, :tnd tht+rt hvcd irtFrt•dicnlt 3 Tn aPprmimate the conditions of human Imrtkinf. Ihe smoke must hc• diluted with an appropriate volume of air before contact with the bronchi and IunRs. Fxtreme conditions of exfxtturc, as to undilutcd snxtke, may he justified fnr cerlain laboratory exT+crimenlt hut, in such cases, cnnclusinnt with respect to normal human experience obviously cannot he based upon such observations. 4. Smoke produced by blowing air through the burning rone tnward the hun end is of essentially the same compotiliun as that produced by drawing the puff from the bult cnd, provided that free burning is allowed between puRs, and that the volume, duration and spacing of pulls is nm mtxlificd S To better approaimale the ratio between particulatc matter and Ras- plus-vapor components of smoke encountered by human lung sur- faces in ordinary smokinR, eufxrimental animals should he induced to inhale through the mouth rather than through the nose. 6. To maintain the conditions described in 2 and 3 above, animalt should he exposcd individually rather than in groups. Suhscyuent to this eonferersce, new and improved mechanical devices fnr e>lfxninR animals to smoke inhalation under conditions more closely approaching those set forth above, have been developed •T hey are currently under trial and adjustment. A solulion to the mouth inhalation problem in mice appears to be imminent and tome prnRtcss in the quantitative meacuremcnt of lung ex• rnture to Parliculates under standatdiYed eondiliont has been achieved Smoke inhalatinn e>trcrimcnts sfx.nsored by The ('ouncil in the futurc ahould have the advantaRe of slandardi?ed cquipment and rnethrxhrlnRy where this is dctirahle for Purrotes of comParahility. Cnuncil studies of the eflecls of inhaling the gas plus-vapor phase of tmnke free from any particulate phase are in proRress for crmqpariwn with the efiectt tit whole smoke inhalation Such invctliRatitmt are Ix•inR cxtendcd to compariamt of smnke inhaled from cigarette tobacco miRlures with those frunt typical ciRars or (.ire lnhacco rnialures, in (rrder tu attett tlrlTereniet rd any kind that may hc ohterved in hinlnRical ataivily. may disappear lhercftxc, rrtt•charut-.tlly ycra•r:ucd tmr+kc mu.l lie dclivtrcd to Ihe aplartpriate tittue tit the c.IX-nmcntal :tnim;ra witltin a carefully enn- I'rr'(lisll(r.eirtg 1'if('t(/rs irT I,urrg (:rrr(•inr,gr'nr'siS trrrltctl tirnc IKtind hdhtwinR formalitut irt thc hnrrtinR 'Prrnc Ihi% can Itc ' ~ adjnttcd by rcPulalirm tit thc drUancc In•twccn thc huntinF nmc antl thc 1he rctkt:tncc ttf heahhy inlact nxlcnts ttt inhalcd /unrct rrf Irurc car ~ anintal with al ltrnl ti.ttr crrntrrrl rrf Ilrrw r:trc and n.c rrl :t cy.tcrn that cinnycnit• hydrtKarlarnt, tuch at mcthylchul:uttlucnc and IK nilryrcnr, 11.1, m m tn a.r,t1k .rPlnr r.. t,l, t"rr rl•n.ttmm .,r r r:tl•InnF n( Ir:ntit ularct crt tuutc hcen mcmirrncd Ilrtwcvcr, intrtaluUinn uf tuth hyJrrK'arhrrnt unJcr trrnrli 13 e 12

.. 3. .a:r. 0 ~"d\ '..°; a, tn !D ... © rinns that insure pcrsistencc nf cnntac't havc o-r,mctimcs f.ccn f0h•wctl by afi/a•arance of canccrs Snme of Ihc%c Iciltniyuc% havc alctr invr,Lcd varir,us tkRrecs (if tissue tlamaRc nr Lrauma lhcae e.l,crimrntal finJntK% have Rivcn risc tn sreculatirnta cnncerninR the rnles that may t.c Id:tycd rrwne pcnerally in IunR cancer Rrnesis by factnrs that influcnce the efTic'icncy of the mamil IunR• ckansinR mrchanisms or by lung damagc ft,llrtwctl by rrRencrative ac- • Irvity Such sfa•culatinns have been extentlcd ill cttmidcratitm nf the lung cancer problem in man I he esisfence of rretlisf+cning factors to human lung carcinr,m:. is cug Rcded by many cf,idcmioltrRical tth.crvatir+ns nf thc human situatir,n 1740 or atl, it is nntahk that only a minnrity trf rcrsnns eR f+ericncinR crtrnparahle rw fatsurrs In varir+us envirmnmcntal eRt•nts tkvchtl, cancer nf the IunR Set drlTerences in int-itlince td the di.case suggesl a rttle hy hnrmrtn:tl aectelirtns in faedisfanitirtn Othcr enek+crine deficirneics or exccascs are supRccicd by stuJics of lunp canicr paticnls at autt+psy ( hrt,nic nr refK•litivc lung infee- tit.ns• f.ctsiVCnt ta't tilt virnve, damage ttt the liver, ehanRee in the efficicncy t f Ih r It• rh „ ' t a ~ c c tt rntlthcGal syatrm, nutruunel statns, altcratitrn rn mutus ct,mfwxitinn. aIlctp,c rcat lirtnc, an,l innrrtncr:rhlc r,thrr fac,tus inc Iudinp cven I+aytht,b,pical chiractrri%hcv, hivc tvtn f.rrq-cJ aa I,reJicpo,-,inR tn cancer ttf the lung 1 hc invc lir.tti„n t,f f., tmn (,r, J,y.,.rnp rn t.ut m„rn:t nf thc IunF his Ihm Ix-cr,mc fn intI-ntant 1,.irl „I I h, I r,nn, iI ti f,r„/•r:un In animil c.l,eri mcnt%• thc t'firttt t,f nrcth.rnt.al lunp 1re,nna im1 ,ir vunc infrcunrtt :rre al rcady undir stndy irt cnml,in.rtn,n ..nlt mh.tl.rth,m t,f .mnkc trr ntItcr :tpcnrs In humans, a new irtveatipati„n uf ht,rrn„nal /acl„ra in Itulmnn:rry cartin<tma has t.ecn initiatcd, which will cr,mhnc rn ht,Khcrnic:J t+h'.crvatir,ns with f,ntr rn,nlem e tamrnation of cndrrcrine gland riasuct frtr hylK•rUry,hy, attt.Phy or other chanRcs ,Inimnl AfrrtleLt for LrrnR Cnrrln(IFrltrArt/ Ctlltrrr'1 A truly adcquate ecf+crimcntil :rl,l,rnith In Ihc hrrt:rJ Itrt,hlem ttf /trc dislw+.inG factnrs, httwcvcr, wt,uld al,f,c:tr ill rcyuirc .clcttn,n nf anint:tl mrKlch suit:thk fur use in sy.lcnt:tuc. Ntcp hy vcl,• a/q+r:u.al uf numt•rttus intrimtc and c%trin.ic factrns that m.ty int'IC:r%c t,r thcri:r.c au•ccl,trhility Aoatiof:wnry animal tntNkl 0unA,l /,rnvitlt ntc:uts t,f In,.,ln.irya hi% t„In/•it.tfly I,ulm„n:rry tntn„r :tt i nuntrnt:rlly.uh.l;n,tt.tl an,l rc I,rralut il,Ir It a I I+rcli r;thly rn .ivrr.r) thllt•rinl .tt.tin% :uul %I,it ic. A h;cJ an,l " t,ht .,I,L •,•h ,r, nl In•n ill ,t tn Itl~ ,I .11 a tr,tl.d,lr IrtiI in i mentinR or diminishing eficcts of superimposed singlc facttrr mttdificatinns Once adrtptcd, such a test system should, of coursc• hc maintaincd with out basic change throughout a long series of screening tesrs in order to maintain comparability. lhesc facts make selcclion of the screening system a rather enacting choice. Several fxnsibk systcros are presently undcr evalu ation or trial ffints and leads deriving from the use of such animal models in system- atic testing must then he employed to suggest ohservatinns that can altprt.- rriately he af plied to the human siluatinn on a clinical basis In turn, such ohurvations might suggest additionll qucstions that can Le explrtrcd with anim:.l m<xfels. • 1,rrng (:IenrRnrt' Phertorrrrmr Among the factors that influence the rates of clearance (tf fnreign matter from the resltiratnry tree is the activity of the cilia which prrtfxl the mucus hlankct with its normal hurdcn of inhalcd dehris out of Ihc lung 1 frc (•uuncil is participating through granls fnr the basic study uf rhis clearance phennm enon and the factors that inerease nr tkerease its eQit;iency In such studies investigators have employed a variety nf animal prepa- rations, including ertcised frog ncsuphaRus, elams, etc. Whilc thcse studies have been scientifically infnrmative, they nced to be su/rrk•mrnted by further ohservatinns conducted under more physiological eondilions in intact mammals. One hyrothesis suggcsts that ikpressittn nf the ralc ttf mucus Iluw may allow inhaled agents from any uwree to remain Innger in cttntacl with IunR tissues and therefore increase their opportunity for action I his hYlxNhesis is slill untestcd as a pnssible factor in careinnRenesis. A dirccl test rniRhl he ptrssihle with an animal modcl ttf flte kind previuusly mcntittned It such a mrxkl systcm, tinder hasie eondilinns, could prtwlut•c a bistnhrpically al+• priate lung lumnr at a reprtalucihle Icvel, the effccl uf influcnccs /lctt dclness or slimulate ciliary activity and mucus flow mipht hc testcd ill htnR ternt etrt'rimcnts with whole animals Such studies w.ndd alai require the ute r,f improved inhahNit,n and mnnitnring tet'hnirluct if rcatrlls were ill fK• inrcr rretahle in tctma rd th,ae and reslwrrt.e I'rtrgrev- in tlcvclrrl.inr .urh Icch niques may rcrndrr Iwnsihle a hcl/cr rv:rhrtlinn nf thc r„li of mnr u. flr,w ac a th'fcn.c : pain.t the atlir,n ill inhalyd npt•nts As mt•nti„nid ill Inevi„us ( rtunttl rt•la,rta I I'!61 64 ). rht• nru, u% II„w is rtrily r,nc :tF•cnry invr,lvcd in thc clr:uanic r,f c%tr.minu. nr:rnrr frnm Ihc InnP Scvrt:rl invi lilat„ra in Ihic IirIJ :t..iFn r,nly :tla,nt It"3. „f Ihr t"d.r) tlcar:nnc attnrlv t„ thr tih.r mutu..y.lcnt. :t~rrrl.in/• :t f.u Lnt•ir r,Intivi IS