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I Microbiological Associates 5221 RIVER RCAD • BETHE::,x, MARYLAhD 29016 TELEPHCNE: I::1/ E5a.3tDo rzy 24, :Sia Cr. ililliazi U. Gardner The Courcil for Tobacco Research 11i0 East :Sth Street New York, New York 1C:22 Cear Bill, I Just a little r.ote to affirm that your concept a•^.d cur ar.ce;t cf the events of last wetk are cor.yatible. Here is our versicr.: a) the proposed stadies are sound and scientifically cc,-peta-:; b) the existing mechanism and funding will continue for a 4-6 nccth period of t:re at a level similar to that proposed; c) during :;is 4-6 month f:;nding period, alterr.ative methods of funding will be st.a ed, evalu- ated and hopefully, implenented; d) based on these contir.;eicies, a ";rar.t-like' proposal will be submitted for contracts 14, 25, and 2B, for evaluation at the fall CTR-SA8 meeting; e) contract 22 will continue in Its present form. Please advise if you are not in agree:x nt with these views. Ncpe to hear froo you in the near future. Sincerely, ~ Richard E Kouri, 'h.0. Director•of Researci ( R:Y:av -.. Nenry L. Schechtman P.S. Please keep in aird that while the meetings wiii :e tzting place '.n four x nths it wiil take time to ieple:x nt whatever c`anees are agreed u:on, ttierefore, these prc;ects should be f:,nced for a-:^:~%m of six -cnt"s at this tir,e. A OfVISION OF ~`~hittakar CTR COF(TRACTS 020285 ( I 11240033 ovrr EXHIBIT NO./6 S. NELSON ... t..+ i i'`e f 7 i 7 04377q

I . ! TABLE CF ceNTEVTS Pac,e l. Brief Surrniary of Progress 1 It. Progress of Smoke Inhalation Studies ; A. Introduction 4 B. C. Equi,:mcnt StatuS f'hysiologic Effzcts of Smoke or Carcinogens In thc Lung i; 10 0. Biochertiical Respcnse of Pulmonary Tissue t to Carcinoyens and Tobacco Related Materials 21 I t E.. Inltiation and Promotion of Cancer in the . t ~ Lung 27 I 1 ~ F. Ref:rences 34 G. Tables 36 ~ H. Figures 62 1: 'Appcndices CTR'-CONTRACTS 020266 11240034 GTR H~~ 04371130

?RCC'._:S ..=PC;,T I. _•ier su--ary of cr -zress. .,. Intrc ~cticn .qui,-ent s:at.:s re;.crt 1. S-cke Ir.^a:2:icri a. Sct1 I, :, SII C b. S-c:c_ r_.i:ors - ,;ar.icula:e serscr_ c. Ini-a1 c:rit2in-ent c. Flc'•I safety syste'7s / 2. AereSo' cenerazor 3. Ci:arette R.7D selector 4. CC-Dxirneter 5. Cryostat and measurer..ents Physioiceic ef'ec:s of srrolce in the lung 1. De^csitier., distributio^, retention a-+d clearance of __ z'.nece^s and to~acc~ related r..aterial. a. './altc^ Scrizcr.:al S-okino 113chine (:~~5!!). u.ar.us^ a ..escr.,I , the firs_ cc....ietec ser __ ,,- thes= ex: .•.:e-.;s are a:pended. 1) _xceri-e,:s 20: Effect cf ;articu- late __ os : c-. :s ac; nu-.:er of c:;are--:es or le^ - _revicusl, expcsed ._ s-oke. 'atais ;.resentisd i .. . s re;c: .. „ 2) Ex~-eri-ents 6, „ 1.4, 17: Depcsi:icn and clear_-•ce of SaP, or u.CA aeresolized in s-cke. Data is presen:e: i;, tiis report. 3) Exrerir,sent 18: Chrcnic exposure to j4C-MCa aeres:.'.ized in srcke. Tnis study o-,as not initiated as describec. 1-(:2-, :21, C_;, s ~piants. Data is presented in t`is repcr:. 1 C CTR CONTRACT~ 02026? CTR HN 043->~ 81

, L ) :x:erir..ent 19 _ffect c~ E9~? in*-a:rac.- eai ns..ile= c^ :l:e ._:ention and cleerc-„_ ,. 1_ oa? aeresalizec i:; s-c:e. This study is ce,olete c-t a-a1',=_is is in prog-2ss. 51 x;,eri-ents 1;, 22. P,eter.:i=- zn- •-8nce CT o'ldd aft°_r i tratracheal '.oC..ia These s' _=s are .:.- cte crid preli^inary res:.:s are ;re- ser.__ i.. t s re=.,... b. S-,c:e ...csure Itc:chine (SZ1•1 I I). 1) Cx;.erirenis 1, _, L: Ef=ec. ez.,,__._ tir,.e ar sc- _=r r_-atica cn par.icalate ca^-s: icn an- cc. c- res_i `.s v+i tin those e_:ei n3o us ir the 1•ia i_-. These s:~-,dies are c~~.z.iete c:1d data i: r'ese.^..__ i^ ...is reDJ'.. 2) l:cute Toxicity Erperinants. 'D ata is prese-ted i,+ tl,is re::cr.. 2. Lung structure and morpticlogy. a. -xzeri-ent A-001: Autoradiecra:y of rrouse lur_ a.`ter expos_ to 14C-DTC labeled cicare:e s^oke. This study is cr.:cinc, however, prelininary res--:i:s are .re- ser.:ec in this re:crt: b. Ex.erine.^.t 1-022, 1-023: Recress ion cf sn.^.:e- ir,d;:^.d lunc lesicns in 3CSF1/Cur fe-ale -ice a-:er '`.c..i_ s-cice exocsure. ;,rei..-inary '-ese st~.;dies are still en are presented .- this re:. _. hcwever, c. =x:=-i-er I-040: 'cxicity r -= aeresc..= i. src<e ar ..is.olocic evalua: - c- .-e ex.ese= a^-2:s. T'isis cC^r1e:_ a'd resu;:s c'- ..isC..:sc_ in ,ris t'epCr:. ~ Jioc'e'• CBi resC ^se of p',:ic.orary ,1s5:.. ..c car...r.:,ce-_ and .:.taceo .'el'e:e= -a:erialS l. ~x.~, :,era 1-C;O, E-CC?, E-00, Seie:-t.•: assays or t`e s ress -esPonse in ,,..ce as a f:._ cn cr a:e and exoesure :e --e racnine or smicice. These s:ucles are ccr,.pie:c and pre .-inary results are preseate--~ ..n this reDcrt. 2 V CTR CON : RAC"5 I ()L.-C5 1 C~"~' ~'IN 0~'~~ r`~:>

2 . Ex-~e-...•ent CT?.-32ACete.-m naticn c- -ha r,ature cf c•.;__-..,,-es involved in smoL:e-•nduce_~~"r4 levels. ( ( 3. _ ;er.reat C7R-8Z5rE ffects of i„ ibi tcrs cr F.;j:, and pro;ein synt-esis on s-oke-induced ~.H:~ ac:ivity. L . Meas-.--ent of orni,-ine decarbor.ylase (CC~, in pulr•„-,ary tistt:=5. Use _- ..-`er substrates to charec;eri:e s-:o:•:e- inducc~ or.icase activity. 6. Cc-• _-, . cn cf smol;e- i nduced and f'.C,- i nduce= 1 7. Carccx;-e-cclobin Assays. 8. Sco,<o- tlated inhibition of pul-onary DN:+ repair in BC,-=1 femaic -,ce. E. Initiatic., and prcmocion of cancer in the lung. 1. Er.peri•:.ent 1-100: Chronic and co-carcinooenic smoke inhalaticn s:_dies. 2 Exoerire• I 1C1. Relative suscert:...iity of C3H/::r, f/:r,f C:.., mice _.",CA-induced lung turaors. COF4 ~ CTR CTR HN 043-~ 83

C, I L• A II. Procress • `-'cre Inhaia;icn St..dies A. I n t r c :..~n . The s-..._ ,..-alaticn unit (L2~07 ^oe:^esda r•as '.;aen ,'.l` _-"a:icnal Since June 1977 ..;a.^. t'e .., carci-ocenes.s a-.: cnic s-:o:ce inhalation s. ies .ere ini a:ec I-,..ae i?i), all animals ware tra.^.sferr__ ._: cf :_::ni t. -. _-... re uni t::as then c;eaned, cec ..__- na:c__ cnc a,es:. _:ed access policy initiatecl fcr ., -.icaard -_ as to prevent the occur•ence of cinfec_icns ir r.- -?ce. In particular SenCai .:r.sl•.cs tee^ _-;.ie _ from tie inhaia:i^ . !.n coi a~crati • .:i. Drs :ack Parker e,d biike~Collins cC ^Cal vir•s .::cine has been tested and found to es :rc;ec- .. -i^e that received an i^., rcnasal ch al1er^_>. Y~ice the inhalation unit areroutineiy vac cinate: and or cccplete seroloSY tes;ing. ne•.a r..ic= bro;:c: t in _.-e unit are vaccinated, su5jected to isolatien fcr -r__: weeks, and tested serologically c:`rir tiis ;ime. Cniy ....e.~ the serological results are ne;a:ive for t;;e cos: ...r-_n ,r.urine viruses are the mice moved ou: of. isolaticn a,C :-, on test. Duri-= :'-e past year, collaborative studies have ~een carrie-~ - -- sc,oke inhalation end related areas ::i:- .,. J. Catcn,~ .. . -'.olrberc, Dr. J Stokely and Dr. 11. C.er..j at C'tl:l; Mr. _er- iorant aid Dr. Joseph Greenspan of ?r„cess and Instrur-en: .Ne,-e York; the evaluacien of stress as a=ac:cr in mice excose= . s-•cke with Dr. W. Essman at Queens Cci:e;e; :e s-oke rela:e: -ni--ition of C?lA repair `.,/iCh Dr. R. =asr,:ssen a; ;-.__ ..:vers i,y of Cal i forni a a; I rvi ne; . nc__- ti , cr l;,r_ cy s^~ofc_ ~:ith Dr. i. wanl at .e _ ..:ers y „- ___.- Carol ina. `Nith CRN'iL an„ , 1. $-J:°_ ...`aia:icn a. _::~ 1 1 3, S_M II C. -=e S~_K II 3 has heen in use since Se:.,;e^.:er 1976. Upon e:•a'__. :n and saccessful ccrple:icn of prel..-.rar, experiments, :ne -=sicn anc ooeratien of the SEM II ° was a-Proved, thus ai a secor,d machine, tne S:M 11 C, Ie I;:ilt and deliver:: .,, r!A. In lonc-teras studies, these ^s:,- -.•:s are subiec:e_ ,., heavy use so that hav:. _ a cec:<_, ^as :ee.. -osclutei•,y r.-essary to assure continuity in tne caily ex=osures. in acz::cr., several exzeriments required -`:e use c- a sec_.... -a_..i-.. ....i le .-.e ; irst was in use. ':x-.eri-en:s H (a~o~~o c?~~ ~~~-c CTR HN 04-~.3f~`84

(. ( ( ( r• 'di `-h the r,_dification in the shate of the hol e,'a ole, t=e ..yiein the rubber cam material r:as raiscc 1/1c , ".;s a--ance-_ :l,,.:eo a better fi t for ,he no•ses of the mice. T-.e ..._.:les ar,c c=- are •roshed daily and then cnce .,•eekly ;ake^. a:)ar; :nc .`--.. etely cleaned, dried and reasser..t. led. Da-s' are re'.lac_da: !ncrlChiy intervals, so as to avoid the atc:-:,latic^. of c:care:te tar %.hich wi l l causa ;he ca-.s t,:r Cracl;- c. =iC':! safety systems. i-ere are two flo;+ safety systess in c:crc:'sc-n a-= _r evaluation on the SE!•t I I. i=.e fi rs: s:s i_ lc=a:__ ,n rcom ari th the S~:1 I I and --•.. :crs ,~e '!c, cf <_-.c< . ai r) after the SE11 1 I but be,'„rca i, cac`.es t:••a __-;airenc unit. If a lacic of smoke cr ai ecc::rs .-- machine, a pressure crop is lost and c^ s::i , is t' '_= so that auxi i iLry ai r -wi i 1 bZ divP. :e: .,, -'•.e a,i irals. 'nis system is also interconne_ted to the par;ic''.a;e sensors and '::hen a predecermined unacce:;a51c level of parcic:la:es is detected, an alarm signals, the smoke is diver.ec --,... ;ha animals and replaced by e.^areency ai.; lf the auY.iii2i~ cir supply is filulty and no flc'.J Is dete'.sd, the __ -o-acnets holding the modules toget'her are reieased, allc•.+incl:=.a mice to breathe rcom air. The flow systems described a:,ove '.ri 11 Pro- tec: the animals r.cm :aulc, exposure equipment, electrical mai=unction, anc _-.ac:e=table levels of cigerette smoke. :1 saf ty system o:as -ee=ec, however, that rrould absolutely prc;a-- ~.gains; a ~.c: smoke (or air) flow after it passed c.`.e -ice. Suc. _.:_. =f flow in the animal containment system _. be causEC s-.o<a or air leatcace in the modules either acause ef ...ci_s -.., saal ina proceri.i or fau'.,y ing fic.r safety device coavoi ;eee c-cc _,s a+as :ir:=- = Process and Ins;rur,e^;s Esc is ...es=-.ly te. - ___ i~.is d:vice c:.._is;s of a..eat s:nsi..ve = s-- .-. ci resoends to fluc .:a:icns ^air '-a -=s:onse of the :her-~isccr is eiec- -.n cally cc,.re--=- :he safety systs- so tna; i_-e -.- (smoke cr a ._ :s beiow a certain preset vaiue, auxi iiary ai w i11 -e civerc_d to the animals. If acxiiia-y a:- is caul v,,' or connections prevent any flow of ai r eie- .-ac-e a to the animal:,a-:=r~a en second celay, the are released cn ,_ mc__:es, chus.ailowing t`e animals to breathe rcc-, ai r. 6 ( I CTR CQNTRP.C~~ 020292 CTR MN 0~-~~ ~ ~~

L. -his , iow -.er-..stor was tested `cr t•::o %•:eel,.s e^ a syste- in ,.hich rachir.e control ^ia;als r:3re ex=osed to air cnly in t`~e anir;al containr.a-: uni; The air !ic•a values a: t^e end of the modules %:ere 550 `,r (fzet ;._r nir :e) for auxiliary air and purse air, arid 450 fr-. for nin chine air. The safe:y system was set to trigccr %;~en ;e air cec _ase^ to iess than 3C0 fpm. The cevice %•:or ed c•ui._ sa:is4ac:eril except for the inconvenience ef loac n5 r•i i c_ ~ri . .-e -oc_ . °5 ^0:1. 2. :,erescl .,,rator. In C,_-=r..ce- 1977, CRt:L delivered e~ ee-csci ca• tcr c__a':le celivering 0.2-0.; p pcr:iclca :. a^.-•a s co, ;zi .ec in a system identical to t`.a- usec c- -~.^ Sc.. I 1 The ..cse cr, aerosol ized chemical car, be con:rci ied by -e ie. :, ire c= exposure or using a' zprcoria:e co,.cen- ,ra-i,,^s ..; -`•e -e-.icais of intcrest. Ex=csures will be ronitorec usir t=e sa:e particula[e sensor used on the S=:•1 II. The precise tec::nic:l detai Is concerning the aerosol cer.erator can be `cc:nd; in the CRti! Progress Repert. Initial ex:erinents utilizine the aerosol yen- era:or will be cescribe: in a later section. 3. Cieere:._ =TO selector. Duri ; z'%:ast year, the sr:ekiny characteristics of ..:c_are:-es of ci,=:r_.., types has been investigated by -easuring -_ .esi=-=-_ to draw, (RTD) with a F,ltrona Resis- tance to Cr~:! >tac::ne. It has been observed that variations i.• avera: " c:., ?."~ ~ ccc . tet•.,een cigarettes rrcm differert car ns c- . s=.-e -e ci ,-e-te type, althcuch :he lar es va- a;:cns c: _rre- cn n i. the _al ciaare::es (-abie Ij .:S _ ..'•Z =..-:-.^.c -. a '.t:een NractlCai Cr1 _-Ine, iarce v ..,, .. '_re-.,rat!;e CiCaret:e .' selecter sc= e e- ec _ ^ _ _ . mac•. .^~(~ „ -SSc.1Gi-:~ Tells) \':as dE\':,: '°C.. _11e c. w d .J:' . ~ s ar. _:a;.:a: _.. c- ..•_ a-_o.atic seiector. The c^c:cr_. cauaes on the RT: selector macnine were set -cr ,~e =oTlawir.g RTD's 1/ery icw (:! ) < 65 rr~ r20 :ow (L) 65/75 -^+ H-0 r•edi u.. (M) 75/85 •.. H-~0 'r.ich N) 85/95 ~r1 •r.2C 'Je~y Hici (HH) > 95 rr:^•• •`•2C 7 CTR CG~`!.T cAC_T~ 0 ~:~'1:.r CTR HN 043~ 86o

L. T`e -... , r.: ..=s I•r Icac^d wi th a:aroxir~at21 v i cicar: ^s The ..,- !n: sclec:ed ciaarettes a: a~rz.:c :- ato.c 1t ci^zret ^s/-ir.;,:e or 1100 cigrrct-2s/- .:s a resu!t cf this sellec:ic', obt2ined the fcz_,.cx'r..a:e r,;.-._- cf ci =r....es i.n each RTD vzlue. LL 13 (< tharn 1%) L lc'[ (< '~~r 14io) ti La (< t`an 22A) k 7E'~ (< than 60%) F - 13 (< th2a . Lcs:. .-.. I y -ac.•ne :~ (< t;an '2Y) 5=M ! I was a1 ibrated for three c' =ere^., ?.' 's anc ,_ p_.r ress_._ eccaireo frc- chese ca:•..-Z :ion: .,,re. RTD P::f` Pressura b3/7: 9.7 mn 1420 78/82 10.3 mm H20 90/95 11.5 mm H20 Afce- t.ie cs!i:re~:`cn oi the machine, cigare::~s~~s with ci =er- enc R'0's ~~ere s-crec at a fixed puff pressura(inter.-edia:= valua of 10.3 r- ~;:C) z--~ also at the puff pressure c:taine= ~y ac;rV_riate r.a: bra; on of the differer: "'s. The res:1 :sere sho: n i•, Ta='•e 2 from these res; :s, and cc sider,-o ,.•:e varia: cr _,Y70 enonc di =erer- _.._-es cr C: ca' .:=s r e_ i., i i, r.e ean '_Orel - e,~,. 'hut •- y~:._r to ~~ ~~anC .._ ver a cJ:a. n a Cc.^.s:c'. _...' •^!'~eof Tt C-.^.s.~.. smckC:...ce".ir_:ICn to tht ariraa!s, :. =are :'es •_- t0 :._ selu cte = C•. _^7 e ___is C' RTi arG a: .'.e E=rrC,.rlate'_- pres:_res. 8 r~C/ Q. ~ ~ Q, , 1,.RC ' ~ ~ (~ ~ ~.G~ CT;~ ~Fl C T R H N 0 437 E 37" I

, 4. CC-0xi-e:_ . I n Oc;^te. 1977, an I l ,--2 C0-Cxir.: t=r :as dei... r=_ ':, to be •sed primarily fcr czrboxyher._nc,,.n d_-_ - .._. - in rice ar;er cigarecte ax^osure. Theex__ri men;t! 'es:.it5 Ce:.°•r"^.'O:! c''er vc.'icus cigarette e>••csar_ leve:_ b2 rcaorte: n a later secticn In addi: -, to CC'-:b e.^.elysis, however, -'-.e CC-=•. -._ autc7wtica::y, and on the :e-ie blood sa-:ie ), c>:y'C'.:clo~i~t (r-zrrenc) Lnd vGiu-° Ox•.cen 'E6 c' ' LrQ~hc_..-:tric r..2ihcd ( ) •:i.9 2^d 1 to 15 s2.^••ples !:3' Gay. 1.• cC_. :c-,, a-es ysis c- ^e va-i_ ic is c~,ser veo i- •.eas ri- CC•- toch r..et^ _, the :0-^•:i.._ter r_ •ires se~•:ar . has iCr.,.. ,•_..,Ticte J', vari.::icn. c'C: \•ery fe1•1 ee'~:•les I;r+ les; c=r:atureti - The cor.c'.-^erttion of hecerin used to - .:!c ;,-Ieoc coi1:_..ng c~_bes,usec with the CC-Cxire:er; w•as~ nc.__.-sed to preve^, ..ouse bicad frcm clottl-; zr. then t!): ir.;tr„nc-nt could be us2d •.rith no iurt`.•.cr rr.odi`.caticns. 5. Cryoste:. An IEC 1•:o::l CTD Harris Cryostet~~•:as purchcsed in :.cc st 1;77 to ens~le'le us to carry out c r 1•C-OTC auco- racicarao~v ex:)erir-ants. lur.c sec'ions`cr 2Utoradic^.r:;Orv pre.zret -=ec, forraiic ,ixed i!ssce are cc-plecely unsatis- fac:_-y if :e suos:=-__ r.= !nter?sc dce. not s^aci`ically 3nd -.vaie!v bind tc allclar ccm.or.ents cr if the su bs :nce is +^_ Gurina t`: prepara:ior. pror;.is. ?rel ir,inary resu::s are s`:c-..ir. in sec:ion D.C.2., ca^cnscratina the I~. d i s 'r i =j :icn and i r.:+ts.=te depcsi:ic^ wic=.n t^.e ju^•g of C- uT .Z e:• :sL i_=: i r, sfn c::e. s C=. RR CQNT~RCT~ t]202°~ 1 CTR VIN 043'7`86