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CiaR NN 042214

inhalation carcinogenesis Proceedings of a Biology Division, Oak Rid;e Nationsl Laborator+, conference :e!d in Gatlinburg. Teanessee, October 9-11, 1"69 Sponsored b}• Vational Cancer Institute and U. S. Atomic Ener;: Commissic- Editors M. G. Hanna, Jr. P. Nettesheim J. R. Gilbert April 1970 .U. S. ATOMIC ENERGY COMMISSION Division of Technical Information _ _• ~..r.-~. . . ~ L, f R H N 0 ~'`-2 ,-2 1

EXPERIMENTAL INVESTIGATIONS ON TI IF l_1=F1=(;TC/I- CIGARET'fE SMOKE INt-IAI_A'1'1ON ON SMI\b_I_ I_l).Itt Ils P, I(11sY ANIMALS f. I lV. IXIN"I'IiN1VII.L IllstiUllC UI Scientific IlCSC:IrCII, AsiUci:ItlUll U! 111C l;rlu,.iil t'1t;.,I.:l1G t,I:1l)U- [acturing Industry, IlamburE, Gcnn:luy ARSTIiACT Fapclimcnla wrlt pcllnnncd .m ham<L:u, luirc, .,n.l -I. i.. ..n rll,ul I., Iind a ph'... 1,1.: Inh.d.,llon In,~Jrl Lu Imnl~ Icun c~poam" I,• i„I~.,.r,. :/u„I.c. 'I'llcst i~IK,wlcflla Wa11c d.:al~nc,l lu IirC:d .dl IiJrllu,n ul Iln. ..,d.a.J, .lu~lu~t, I..-fn•a alllul:C cvp.,aulc w Ihal lllc cIlc..ls .,I I,uv.lvc ,lu.,l.inl; .,n .mw,.1l, ,..~J.I L; ...uq'.u-l Iv1111 Il,t clf:LI. ..I.,cllvc aiuul:aq; un lu.,n. -Ihc lu'.ICII)' ul -,c...1 ....,J.: pu..,ula, aucll .u (I .nl.l nli„liuc, :IUJ ll,e iulnl,Ad.illly a.I .Illlc,,:ul . .I.,...u. lcele ulvc~Ui;.ilr.l; inllucllaea nl i~,l,h.ll.ny lI:,lu.:u.y ,..n~.,. ..i r. lU'l...)Ir.an.l IcuL~.i)•Ka, ai~I1,uU,l.llluu IJ1C, hu.1) H.CII;1,1, .u,,l Lanl \vCIC al.,, w-,wlyd. fJu,':uvtl, lvc li.,v: lu..l r~p:ninenLJl, ..L~ull II~, .I_aull:..ncc,d Ihc v111'L,nCCi lu IIiC lu. .r ... ,.II ... ,14: ,,,cIJLUllmll ul al.iy I,c,l.,d I,I lalClnul(CIIIc ea,elu..leuie sul,al:M.:ca. Illal..lut•IC:d .li.ultrt:. III II.C n..l.,,.~i,~ ) u.~,.I ,.,Il.,lunt; Iung-lrnd ItJ. It)n,,~.~o...,.. p.u/1.lca, v:c li.,ac Illt.l 1.. c~pl.du II,c .d II... ..I, ....~.~~, .1. ~, .. I..u,'J. IAKII;-ICItII CXpCIi1llCllls willl syll:lll gul,liu I/:nli,l,',:., ..,,ull-lcl1:1t Ill 0111 hhuIn• IIHic) Ill I'1h1,1 'I ,IClnuniU:IlCII J IcIJGUl.:IllIc ..II•,In.,llul,~ Ill Illc IC)Illl:lluly ILICI I.,IIU)vill(; 71111i11i:411i~,u ulIlilluLlllli/,c,. 1,I,~/lun:nll v.•CIC IIICICliqd uallcrl:ll.cll tu invcslil;:llc Illc suilallilily ul III,:.; Il,nl,..J.a; Ila Icsling ulllcl iJtluilieJ ur susl)cclell l::ucinuEcni.: iul,sluncc!:. 1Vc bcp:/n (iur Inll:ll:lluull cxpeluncnh. :Illuul 11) ycals :Ilrn.a Syrian h:unsters wClc cxlwxll i/) /;ilt,:nell: snn,l.e Ily lu::.n. ,I .1 musd.illl; Clrulll,el Ilcll \vai rrl:llivrly I:ul. ' c :lull ul Iln I}'llc ,Ic.'lll„rll Ly 1•... ,Il,.:,.1'' (I•11;. 1). s,lun, lluwevel, wC IC:IIilell 111.71 Illls IllullCl by~.IClll I,,, ll~l,.il.,u.,li a1'I,IICs ICSUlls Ill a vdlY Iu4V CuI1l:CI111JIliIH1 UI, YI11„Lc ill Ilil; ':IGIIlII1Cl, I.,i,lb-Ul.lil~ .II IIIC IICIt11111111l; l11 IIIC cXl/rlilncnl. I11 allllil/ull In Illc I,I- I,ulcllt p:I1liClllale5 in Ihc sn1ul:inl; clI:unhcl, Illrlc :u.: scvcl.ll „111,:1 Ln:l.,l~ lu becunsiJtrell, sucll :u Ihe I,I/s,if)Ilily ul nus:ll'Illll:lliom :lu,l IIIe I,,,d,.nl;oJ „I :Illele/l hass:lgc u/ inll:llcll sInul.c IIIuuugh Illc n:ual i:rv11y.

X 4 r.Ale wr UUNTENtvILL PII;. I 1 u,/, w.,rLinc wilh a largc vacuuln chambcr designeJ for 10 Itanutcrs /1'lulll l'f,C111,1•16's1 I Ig _ ~ Sulod.intt ul•L lu... • Uh c ulum.ltte .uloAe/:/ir venlil:l/ion, dcalenr.l I„r , 1,~1~1 Icnu.l.•n it) pr I,..,,1. ,. ,II ", I„i.J sluuking syslcul"1. (II.,u, I),ulcuwlll er'I'I'(TIS t)P c1(inltEt rt srelul.l IIIII. l. lll I1 Iu ulu ulliniun Illcrc tvelc Iwu Ilruldellts Ilcll II•IJ 1,'~ La .,dkrJ: 1 I 1 JI'sy;uiult a swukiul; dlalullcl device Ihal Inudln:cs .Intl ull~ 1:. Ilic :uuul.ll, I,q-l, r„n.'I nu,l liu(ls lJl' snlUkle witlllllU entl:ulga(illg Illclll UIJIIUI r.%I,i11iU:111.11 ,_I.ulllu,.lu; ('t) puisuning :ultl niculinc inluxic:l(iun wclc csl,c..l.lllN ll, I,1 (2) pruviug, ill expelilnenl:ll slullics, Illc :lssinlil:llil,u „I ,In,.Lr u, III l:,llu.llw~ org:uls autl Ihc ellccls uf slnuhc culnpuuculs Im Illc I,...pilulul, i)wing llle 'llliltl (1uaJrellnial ('wllclcllic uu (•uu.••I, Ilcl,l lu Pclugi:l in 1 965, wC6 reporletl tesulls ul c.\IlCsinlcnls with u Iwv. l) I„ 1.1 wl.,l.ull, nl:l.:hinc that was.hasctl tln Illc clusell syslclu (lypc I).,nlen,vlll) I I. uull (I'ig. 2) tv:ls su snlall Ihal lllc Il:lnlstcls welc eal,l,,.:ll l., .1 Inl•1113' ..,u,cnlr:llcll aUuuspllcrc of snwke and :lit. 'I•Ilc ln:lt:hinc I,11„•IJ.•J CyClc with condiliuns slantlculliied :ICculllinl; lu Illnu:ln sulul•nll: I<r ults ul Illcsc cxpclinleuls Icve:dcd suluc Ilallilluly prulilclaliuu., „I IIl, In,cII,:;J c1/ilUcliuul :Is wcll as Iuclalllaslic ch:ingcs, intluJin); :llypic:ll nurl:l .u.,l '•rVrlC I.t.dllcraliun ul the ullper P:ul uf Ille Iradlc:r, Illcsc Ili~lu(~alllul,,, n.Jl, .lu. ll. sc:nlrll tu bc cotup:lr,lhle willl precautceruus ill:ull;es. In IV(,S we Ileg:nl exllclilucnl:ll sln/lics7 un 11. .' I.i. L.c ,lull nn:l:lhllllilu ul ('O (Fir. 3) in Ilillctcnl slnul.iul; syslclns, wllil:L ~.., ,.JI•,I I11,: ':uIJ Ilk: ulxu•' syslcnls; Illrst stutlies ,lulwcll 111,11 :ul cunl:lin% lilllc lisk lur CU illtuxu::lli.w. AItcl Ill0nllulrlll ,u .'.nl.,nnl 111c loulu•lUuu I.I ('(1Jlelnul;lullin :nul Illc :usullilaUun ul" Ili.:uliln i,u•. Ulr I1u,,•, .lu,l utllcl Ilig,nli I D0 •ul IU l, '/ - /;. / / I lIJ au IU u U II • .. . I I :IZ, 1 1f.11 , . ~.• 111t. ) In.,..u•: ,I 1/ lln•,.n.+•I•~Ln~ ( u, Il,c lunl:, ,I I,.wnlrl.

OON7ENWILI 392 EFFECTS OF CIGARETTE SMOKE INIfAI.ATION • . 393 60 so 10 0 T StoNf'f t) S6ttln t NG 1 2 1 itt. 4- ItcJu,tiun of ('O hcrnoglot)in in the blood of smoke•expoced hamsters: in thc ptc.cu : (d Ilc.it air I-•-) or after addition of Oz to Ihe otfcrcd air voluinc /-/. ll touu I)inAcumdl rr vL~1 30 0 120 /- (7 ll z 10 x x StOPPEO SMOKINti r A x x x x I.ig. S- Itcduction of ot, ~,line in Ihe hamster lung after smoke exposure. x= mcan front lttrce anintats; •- tlieall frnlll six a/11111aIs. (I'rt1n1 I)nnlcnwitt er n/.~) in different inhalation systems (Figs. 4 and 5), we developed a ncw smoking machinc, type [lamburg. This machine has sig,tilic:ntt ativantages, especially for lung-tenn inhalalion studicsa (Figs. 6 and 7): I. It is possible lu expose animals to citiarelle sinuke liver a lung period of lime. Al present, we Itavc ltad nwre than 2 ycals of cstmicnce wittl 1Lis type of Iuaclline. 2. "I'Ite maclline offers freslt smoke in umt:eutrat'suns and quality that rctnain constanl during Ihc cuurse of exhcriment, and it guarantecs a uniform rhytltnt of sulukc flow. Alorcuver, the distance bctwecn cigarette and animal is kept as small as possible. Fig. 6 - Snwking rnachinc lype ILmthurg ("open swoking systenl"), edaptable to either 10 Itamsters, IO rats, ur 11) utice. The concentration of !he stnoke/air adtoixture is adptstahtc, and Ihe puff vulonle is Lnlonlatlcally controtterl. (Frorll Itcckich rr af. "0 1

394 DONTENWILL Fig. 7 Animal-holding wbe of the smoking machine type Ilamborg. Within tlre lubc arc iuuu •I d/z.luun equipment and an opening into the smoking chambcr for Iht nuec dud ,u•,uIh uf Ihc hamster. 3. I'ilut sluJi.. Ilave determined tthe smoking cycles Ihat assure tlle most el7eclive Josc :II Irllvest toxicity (Table 1). 11 has becn possible to dclnunslrate cxpcrilnentally Iitr luinb concentration uf ('C) (ref. 9), which is a Iimiting I:Iclor in lung-tcnn inlt:dafiull tests. Nicotine, on the olher 1)anJ, is tuleraleJ even in high doses, but the Ilu,r tulcratell varics according to tlle species tcstcll. 1'A IS I.I: I Snwking ('r, fr. I)urarioa ojEx/wsure to Smoke, Nmnber ojCrgorettcs Sn)okcd, m,J /./)StI of lla)nsters Snroking Reference ('igarettes irt the 5rnoking Alaclune 7'ype llaprburg• Jmuklnl; : yric (rifaretues/day) 3 X ((1 3X3u 1 x 60 I X `/11 2 x 111 I/ullc.,lcrl cunltuls •I•rom Ilcckzch et al. " I I:ivc days a week. Ll)su Iwlc (w.cks) U.,y%l uicxp•,mulc lu .ulukc/ uuwbtr of iil,arcllc% sutukcd 1.5 7/135u 34.1) 17n/15,3n(1 39.0 1') 5/ 1 1,7(1(1 )Il.ll 150113,500 >34-(1 > 170/ 10.200 n2 >S? 1/ i EFFECTS OF CIGARET'rE SMOKE INHALATION . 3f 4. According to inhalation experirnents carried out with aerosols and investigations on the intake of nicotine when the nose is opened or closed, it w dcmonstrated that one cduld count on thigl) smoke assin)iL•Itiun by selecting : inhalation apparatus that is technically appropriate. When experimental cigarette smoke inhalation in animals is to be cornpar( with active smoking in man, many factors )nusl be carefully controlled al considered to assure that cigarettes "smoked" fay auimals copy actual hlnu; smoking pallerns as closely as possible. In studying the tespiratury frequency of hamsters in the "closed" and in tl "open" smoking systems, we' o found lhat respiratory frequency as well 1 respiratory volume is increasingly dirninishcll during long periods of inhalatiu and (Ilat this effect is especially pronounced iu "clusell" smuking systems (Fil 8). Accordingly, Ihe assimilation of smoke decreases. When animals are expose to cigarette smoke in "open" smoking syslenls (Fig. 9), respiratory frequency not affected to so great an extent. According lu uur experience there are tw lin)iting facturs fur inhalation cxperiments: C'O-heuwglubin concentration an respiratory function uf (Ite test aniulals. Fig. 8 Ilcspirulury 1'reyuency of hamsters aflcr slnukc c\pusurc in a"closcd system•. (lyhe I)tlllttll\VIII). Itl'nel:lory f1Ul) of respira- ,tiou is ,cen allcr beginnirlg of smoke ex- pusule. (t-tuln Ituckcr aud Uunlcntvill10) t

JJU I'~ IY6~'IYbY~ ~'~ r11~l4tilh'~ 1-t5 9 35 c'1 101 35 /... t 11 30 DONTENWILL 4 5 70 106 1-ig. 9 Ite,ln,.l,,,, r,.,locncy of two Ilamsters after smoke exposure In the ..ul,cu au„ll,.g ,, -1.,., ' Itcduction Of rcNpiratory frrc(Iueucy aflcr conlimlouf ,.,,. I. -i'. I)uration of amuking, 17(1 ur I(111 uun. Numben uu,J g,.el.ll IuJI,.,, ,_.LUAuly Ircyueucy per u,inutc; Il,ose on Ihe oulside Iud,c.Jc 11411116,1 ,., 1.1 ... , In 0garetlcs c.,ch). (I:ruul Itucket aud Uontcnwilllo) 1YIICn rc,puns,~ ,./ ,,I,,:linlen(al anim:lls to l't7 are lu be interpreted for'.. j ..lupauNun wttll Iluulal, „,lu,nscs, it must be stressed that CO-11b valuef ;T I,uing relatlvcly rlul, 1.1, lu animals llllrlllb passive SmUl:illb' are never reacllell " t,l 111.111 after a,:uv%: slu.,l.,... : :\s we have recently reportetl,' 1 the number of ': ',}•11ur,cyles is signitic:l,lll, u,crcased after hantsters are exrosed to cigarette ,1„1.4.c; this ioclcasc I,I.,I...I,ly cau be explained by a rcduction uf lho ,I 11 1');ul-Iranshurtilll; I,clw,l L.I,uI. I luwever, no rclnliunsltip betwecu smoking ;? ,lu.l ,Illcl:,liun in wllitc ,nult cutlld be tlclnunslraled (Fig. It))• 'fho ~ bclllnlent:,tiuu rate in Ilaul.l.. , is uut accelerated (htring exposure to cigarette ' auu,l.e. I/rvestigaliuns ul Il.,lil l,•,:il;llt dcvclupntenl" 1 distinctly intlicaled that '' ,Ilun,,lls cxl,usell lu cig:uctl,. e nuke reacherl, and maintained Ihroughout the c;I,~nlnl:l,1:1J peliud, nNiLnill .-Ily luwer body weight Ihal was:us,>LiateJ wilh :' Jcclca~.c.t I.,ult cc,nsun,y,li,.., I.i Iuol, detailed inspection uf growth curves ull/lc:,ics 111•It ttays willl,/nt :.1-.1 lut; :Ile associated witll a slightly increased body lvcilall I I'1,: I I ). 'I-h"c cxll, 1,.. „u :Il,ain slluw Ihe a),rclitc-inllillilinl; cllect uf EFFECTS OF CIGAHE•fTE SMOKE INIiA1.ATIOrI 7 I CONTROL hi < m {:, lV^ 1 I; 5 t; .: ` ~. t:- 4F04 SIAl7T 10 ;.Q I,,./ I I' ,d.: Juyo Flg. I11 I(ffccl of clvalellc 5u1.,Lc ..u c11'dn,,, Ic ...,... 1 .., I...., ..,. 11 „w, Nc,kicll :,u.l 11,ulcnwllll' I ------ C.(BlTkOL ANIAtt.l J lU ~.I1,Akf I Il ):G.)I;1 . 1. ,' I,. I..... --'-' Li.rut,uHl- I,. ..L,•.ul I II I , . , L..-, , .1 l --. I.---- -t----..I -.I ... I I I I I -I..__._.I-..-. I-.---~- (/Uy! IJ " l) •1:1 la) /', '.hl I., . 1,'1/ 1:', 1:.11 Il.l) 1/ll fill fill tm Imt mn mm mm mit nm tmt tlm fill mn 14 Iml mll n10 fill fill Iln tm 11.. 1....... 1:1'. .I.:tclupnlcld ,t buJp

3`.ltf DONTENWILL TAU1.1; 2 .Sttrdk•s u%.l dnptive Z prrtu,rertcsis itt Str)oke-1:•x posed Ra ts • liuw ul iu.~....,Lu„incl I',ctrcatmcnt luu 12 lixposures tus,nukcuf I+,u 3 X)tl cig.trctles daily luu Inicctiunof •li) tug/kg rigaretlc smoke cundensate i.p. IUU )t1 mg/kg 100 1 bcntpyrene i.p. 160 1'arlial rescclion 1611 ul Ihc Iivcr -Time of p:unlysist (uliu) C'unlruls( Difference ('f) 146(414) 177 (4(4) IX 234(19/2U) 335(IX/211) 31) 232 (18120) 388 (27/3U) 40 79(13/15) 140(11/15) 44 44 (15/30) 143 (2(1/30) 69 23 (6/lU) 87 (8/10) 74 262 (9/9) 192(819) 36 700 (X/9) 29X (9/9)Q 1 SS •('totn I lunlcna•rll Ci at. 1 T I, I u wb c nt+r+. ~to1 C. I 2-aln l nu-5 , 3Nund,rr, ui p.nrull,c.c> indi.:atc animals willt paraly5lS (1Cr total CX(1CfII1LL'rltJl 8r111119ls. 4S(wu+ upct.,ti..n upcning of the pctitoncal cavity. -I'AItL)•: 3 Strr.h. I, .•( I.laplirc• Zyrnugc•rresis in Snwke•Expuscd 1/arnstrrs• I lu+c ul to.~atul~+nln~I r+rlrralmcnt Uupf (.);1 I1111 I.' I~t,usutcs I(,l/ +,• .,,+ut u( + +n,q:,rcllcs 1l1(1 I„p, tt•.u ul- •1uuuJ1K . Ir.u, n: satokC t:un.l. ,,,•,tc 1.(t- funuvl, brnil. I imc uI p:+ralysis) (a+in) ('u,dtulsl 11i11iacncC ('J ) 49 (16/2/)) 67 (1 X/20) 27 I2X(15/2(1) 16i(IX/2(l) 22 dX ( 15/2() 57 (13/a1) 16 49(ta/21) 57(13J2/1) 14 51(X/1(1) 73('1/111) 311 51 (29/30) (,1 (28130) 16 •I+um I)untcnwdla•ta/ `' ( ! EFFECTS OF CIGAHETfE Sh1t)KE INIIAI A f Ivt•) . ' 3( i Slutlit:s uf :u)aplivc zy'nturtalcsis lulluwiltl; .null.c cxlt++surclt Irltrvatl 1h: II1e futnlaliuu u)' hytlruxyLlsc is iulcnsi(tctl a(l.:t wtul.c c\l,umuc ('l:dtlc. .'2 au .51, ant) tllalt ttlc hytlnrxyhtsc is :tlllc tu cluuin:rir a1+,ulaU+' 1+V,Ituc:ubuns. Ou uhsetv:Uiuns Ilave :dteatly llcctl cunliuttcll I/}• 11'clclt rr +d. +' nl lct iuveslig:tlitw in ul:ul. As I:ulin t.'t ul" t't have (ruiu/ctl ultt, Il+k Inu,:cs, uf cllr+litrl(iuu, whita ubviuusly funt;liuns vt;ly wt:H in Iruttsltas, nl,,) IIl.; tlcci;lvc (+n Illa cifcct u t::rtt:iuoltrnit: sullst:lllccs. In uttlt:r Itr keep l+lxicily as luw as 1wsNil,lr Jlnlnl. inlr.Jaliun cxOclilncrtls scvcl:fl Ics( t:ig:lrc(lci a'clc usctf uu tlilll;tcul :,nuuc+I .I+ecic, lu (IctCrntinc llh' acutc :ultl chruuit: loxit;ily ul' ccrtain sul„hr ,-,ulpunents." 7'hc reslllts w, t,bt:linc,l have a prtm:uy bt::lling unly un I11e ;,iunr+l cxl,cuttlcnl bcc:nlsc, a Plt:viuusly statctl, a('U-tcslturlx; uct:ultinl; tlululg I,a.bivc Sluul:istg iu auiut;lls i: ncvcr rtt:It:heJ tluriny active snluking in ucln. Idcccrllclc;s, we trust tllat surtti rt:sulls fruln (lie cxltctimen( may be applictl 1„ Ilt;w. 'f luac toxicity sUnlit:: tlculuns(ralc Illt: fulluwillg ('I'ablt;s •1 aud 5): 'I'AIS1 1` •1 C'untparative Studic•s urr Tusic•it)' u/ 1t./, r, nr,•.in.! 11uJi/ird ('i-~arrttrs (Srnu!-tng.11ac•ltirre l)yrc llarnLrirg/, tritlr l'.u rrrular 1(. ,:.n J to Srvvirdl (irnr an+/ ( U unt( .ti'iru(inr ( •unr"nt I ,+1 N,c ( •iyur ctrc>' - 1 u,,, ul c\Swituc lu amuke until dcath Vu1. Ni, Miuc l Nn,nbc+ l hnuN Typc uf cigarrtlc ( Yl eunl.n u A(ousc l`') uf S+uuS.t pu11o pc! Il +msicr Itat 1i/c ((SAl (uttJcig.) . pl.+rrl (Ill) . )as) 1.GIauJ~rJ) ` 4.6 2A5 lu It+-I IL 76 15 f 1( hutnidtty < (1; / 1. S >.t,U I++ I I I S! 4 4 It• I F 1csltdrlcJ will+ 5,1 U.LX Iu I 1L 6•1 3X 7lP„. cthanull 1(il ((i - rcconstilut.,l 7.7 (/,X2 a 15 X H 12 Inb.ti i n) lt:t (f.(; / t NaNt)~) 7m u--17 I [I M /. 14 l'~ (h t,d,ara:a .t.•ut.l 1. S u, tt, v / I !-1 -1 1 1') I n Iwcuu~litutcJ 11.1 u.6•I lu !6 I I I I 13 lubrccu ttwu I.s ) K(1'.+ L:harcual lillcr) 5.1 1.1,8 IU 1 14 17 25 19 A(I'.+ ]ICId(C tilt+:r) 4.2 1.11 SlU IhU 27 .Sll 411 I. +('amtm,lgc lille, 4,6 I/ lu 1•17 158 1~ 1O2 11,64 10 49 22 17 1 1 0 Ui 2311) Iu 20 1` •1 It k h l v % r : ru/n c. tc rta . A t., .. . ,. n+ u, .... ,.

DUNTL-NWILL •1'yp` uf/ ly,Illlu 'S,:c I I.r I 1 TAI3LC 5 Comparative Studles on Toxfclty of (Ygnrlllos" Nicotine Number conlent of ot'sluoke puf('sper (mg/cig•) plate "t'ime of cxposnre to smoke until rlelithl Itmnstcr Rat 12.3 2.9 20 53 9 '_.9 20 100 26 I_',l (1 20 63 15 1?It 2.5 20 67 24 (ntin) " htOllse litOutO c (I('1) (DAL9/cJix In 12 19 21 16 Illvestit;;llittns of Iile gaseotls arul Ihe particttlate phases.y~ inJiL,llo 111.11 u,. /( 1 Icsltttuse is equal in uniln:lls such :Is hantslers, mice, and .:;~ 1,115. !. ILlnul.,, .,. 1,,,1 at uIl as susceptible tu nicotine as are rats and certain . 51t71ills t11 Ini,Q. /: i. It;3I 11,: willl cl very high ('O level are unsuitahle for chronic lull:d:lliuu cxIl,allll...,l, 4. 1 I1c1,: S,..;iu•. 1,. 1•.; •1 suluur:ltiun effect of CO and nicotine. These findings, .is wcll as :dl ltl. .••I1. , 1.::,ulls nlenliunctl, en:lblerl us to detertnine all oplimal .I,la•Igc Iol ulu inll,d.lll..,l expeuluenls. Irt lllis connecliun, wc wO1rld like to I„unt uu( II1:11 Illc ,t.. •..ly.lliltiun ul nicnline, fur exanlple, in llle h:unster liver, ia signilicanlly •I,:cclcl..1.,1 :11 ctrmp:uetl with tlle tlecolnprrsiliun uf nicnline in • ulc rat livc( iu r•itro. r' Actually, Illc Il.u.nl. , w,ls fuuntl to be the most suit:lltle test anintal for il,ll.lle(iun exltcunlcuIS, l.. I•.1,I hec:ruse of ils resisl:lncc It) Iwllntmary infectiuns I,IJ its :Ibilily Itl tlc,:,ll,.l..•,.: niculirte, and especially because uf llle type and L,:.Itr.m of tllc cxllcll..l. ,.I.Illy inJuccd Iunl; tutuors. In this cunneclion, wr,:tugaliurts ul J.11(tlllrl ..., lo Iiellultl anll 1)unll:lnl,t7,`n :uul Kux:hnerlv lu.ly llc cttctl. 1lcl~lre: Ihr: effccl of I•••., L:rnl exposure cm the respiratury tract of hantsters is rcl,t/rletl, a I•ew ex:lnll/l. ..II„ultf lie menliuncJ lo denlunslr:lle the primary tl.::.lt:rt of cxpcrilneuts. 11'. .•., vcry grale(ul to lhe nwnogr:lph of Wyntler and II.Illncnln,lu wllicll l/lacc. ..• I,i/lclallle emphasis on (lie rlevelopment uf a"less Il,n/lllut CiZ:uettc" ul "•1 h•11. , Ilr:uelle" mtll raollecls su nt:lny cunlribuliuns of nllllulll:llive Jisctls;luns I.•. .,c shecied :Ire:u. Our own cuJe:rvurs, Ittu. have llc.a/ I/unl,u lly :unlcll :11 tlcl..,,,,l.onll; Iwsslblc risks uf tub:lccu slnuking, as well as Iln,llnl; :, w.ry of cllnliuaullt ,., .:Jucing Ilannllll substances. lVllen ttbservalions of a rcduCcls Ilnnttril,cnic acll. . 1I1 slxlnlnt nilratc :uttl ill'ccrlain tolrtcco shet:ts wclc lalllll,llctl ', 11 \V.li h111. 11; .ulvisctl that Ihis :ultlilive slluulll ntH lie useJ LI-I LC IS ul M,+ f. . • . . . -"-' _ `••'' ---- . n-....1._ ... .....1 l/ ..............." ....1 d:.n, o,. .,. ......... .. 1.... ..,. .. .. .. _ . _ . . . .. Others have repe:Uedly puilttetl uut Illnt tlle arklitiOn 411 s1lJlulu I/lu•11: lhe causr ul :w increascd li/rntaliun :unl uplake of nilntsaluincs. iinswer the tluesliun ot' whether tllc atllliliuu ul suJiuln nil/al.: t•. II•I1;1.,:,( possibly inhiltils lllc Inutoligcnic activity unly in Ille :uc;l of lll,; ,Liu, tn t•:lu,(Ilcl the activily ul Illis :ultlilivc is enll;lncetl in IIIe Ilull;s. I'llc,,: rl ll,ut,:l.lll'.Ilz S'provided a valn:lblc inlpetus tuw:ntl' dcsiEning :m inlllul lwll Iu1,J,:l sy..l, ul Illr , ;enimal inll:llaliun sludies un lot:d sluol:e. In our total progr:nu uf expelilnental inllalation slutlic,, llw plill,:il,:ll slr.:,; was given lu Ilte :usutnptiun 111:1t tullacw sluuke Inust Ic•IJ lu a Ileiiuile IC:I, Liun at syute place un Ille lesltir:llury Iract wllCn :Itlluiuislclctl iu :.Ir1li,icully lut!ll tluanlilies lo Illc aninlals. 'flle tesliug of lllis wull;ing I1~'I•.111 ~:.1, a,:culcJ I., us urEenlly rtccess:lry. Irt llle c:ue'thal sut/uL u.ual liltlauuu I•::1 .:,•uSl,l,a:,lllc qualitative ;nnl lpl:ntlil:llivc Ietluc(H/u ul Ille SIl1ULC a+~+1u111.i1,:J, 111e I,::I:Uuu V ;ir ntusl altpc;tr iu Illd arca of rtiu d Irusagcs; ul, in Illo L:.c.c tl-l ,ul ll. i:nl I/;ll llclcs , have p:nsetl llle nas'll regittu, Ille Icacu,m Inusl :Il)l,.;,u al .1 /~~„Ir Ill:~t.lnl I•Ia~e in lhe resllil:uury llucl, i.c.,:ll :1 Illacc %•:Ilclc Illlyslt::dly 11 , I l•I •l ,14I,..Illl,luun is lo lie exl,et:lctl. In lcnus u1- llle su•calletl slu,a.c-Irus•It! , e Ic.nliut; I1"t1, 111, n•.Ilil 1., 111e Iung peripllcly, wc Il:rvc lusltll,ltically luvcbulsllC,l ,cll:d „• II•1-: ,•1 Ilu.; I.,.luun uf 16e leSllnallny lrtct. IIiSIolugi.:'ll ullsclv'lliuui lu Illr 1.." I•1.•In:lli, :InJ alveuli Ieiuulilnled r.aulls wlu, ll I1:1,1 .IL::IJy l,:.vl L; u, ullf u(Iltr InvcsUt,:Iluls. 'I'llc n:u:d c:rvity t FIt;. I'/ :IUJ lllc ..I1Illll,,•I n,ll;ullnl:lltay nruCua:d t.leulVcs :IS vvcll as cl1i111cl1:11 luel.llllusi.l, ul .unl~l.lls v:llicll 11.141 been cXlrustll lu slnu6e fur a luug Ilnlc SIU.JI lyl.l• .ll I. Inll.n l.l:. a'.:I,: (,luutl in Ille n:u:d ccrvlly ul Iwu aulnl•Ils cxlnlx,I 1„ ,luol.c I- Irlt• Illln:. ,\Iveuli (Fil;. 13) slnlwetl :I ctlncenll:lliun ul uelcl,ll,ll:ll;cs wllll I11 •. ne.ll , Illlll+ llal inclllsi,nts, which welc tlclinetl Ity OIlul2 :Is "snlukc-cclIs. . IC•:! .Ilwll:lls I.Il.lt werc ' explrscd ttl snwkc f,tr a luut; lintc sln.wctl SIn.lll ".1,1_rtlllll;ll.nd Icsiuui" in Ittc Iung.l" tliltcul ex:wunali,m ul linlllut:S ul Illc u:c..l ,.nll) . Il.u llca, culd Iuu}; is ttnwrrranlctl in Ihe cunlcxt lil 111.; lul.ll Icsull,. I I/, SIlall ntll lliscuss Ihcsc liulliul:,s in tlelail hul will lurn, illslc:ul, 1.. 11.. I.u;u;, .vllidl Il:l,l,:li I:u :u we have wcn, the nluNl Iclu:rll.allle allcl:lluln•:. I•ul cllrunic snl(tke expusutc, I•Itl Il:nuslcls ".,.1c ~elc.llal. As shuwn ill Fig. 14, 93 :ruilu:ds sulvivcd snwke exl>usulc lul lu+•IC Ill,ln Itl luunlhs. 'I'llc ligtue alltrws nul unly an in1e11ncl:llilm of Stnriv.ll .I-u.,-:• I1l 111: cuulse of luug-tenn cxpuNule Ilul alsu :I Icullr./lal nltcllnclullun 1•I ..,,rll.ll/lrtl Ilislulugical cll:ingt:s. ' In In J.a lu Il:rvc Ilcttcl Cunll ul 111 II1..• In,hl. •.I we tlilleren(ialetl live stat;cs ,11 cllilllrllal cll:ull•. : nl I11.: I:nyu\ I I.II~L u l, u,u lr Illc .lNui I;f Ywnl,r /i!(l1/-l/r1•t' lll 111e A1111C11 l',11,:CY I11,111111,:"1 I~.ill~..l.ll')'~ .li Ital'llall~. As rll Inlluau Ilalllulul!) , a Jlllclcnliala• ••1 ++.Ilr,l I,;eutlue1/i111.liunt:tluns ch:nlgcs :ultl lIlc Iluscl ,/1 .:Il.:inulu:n 111 Illc I.w, 1.; 1S t•cly llllIicult. Alrylme willl lunl, cal,Clirnce In Illn Ilclll .lull wlll. IIW .11„IVC l.l,:nlltlncll :Ill;ls n olller 111i I.llulC .Ill IIIi\ 1I11)jCI I Ill Ilnn/I lll.1l Ili,: t•/ll. lIun (UI In:Ilil-llLlnCy ill 111e 1

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