Tobacco Documents Online

` a / 1 5IO-RE SEARCH INSTITL;TE, i.v:_==,,-•.--c_) I C _;;vA':ENUE ETTS ~'di '/ e1<'e-3t Dr. F.oSe r: C. I-iocket: _=-ssociate Sc:enti:ic Direc:or The Co,:=cil :or =otacco ?•esearc: - :. S.A. 11C :ast 5 3 :1_, Street N e : York, N ew Y c r k 1CCZZ Je_r B __. -'lease --=- __close_ the origi a_ c_ c_-.:za1 rep:_: cc _.e s..:c;- eatitled ": :: i>:panded Sr_=-; o- ::ouse 5..... ~x?os-.:re _ ~':a_:e S=. oke Co^.densate Co::_:ared ._ S:::r. = a:n _: : cu cc:'__ recaL, : have :nade se:eral :i^:es ._ •__.... rrports c: ._.:s s:ua,:, -'_e last :i: :e __n ry i_:ter t: y:L _ a..-ar; _ , . -.. o_` :is re:-r: are be'.-; •_... , a •- r. =ioyt, Dr. Gar,_..__ an- Dr. Lisar.t:. ?• ,:s ....._ t: _ res:.___ ._ -- _.:eres_ ._ .ne gener-l pub_._ c:- -- a =::a= oubiicat:o:: ..• scienti___ > cz h:.:,e tt:a: Tilr C~ nca f~r _coac_ suc:~ ca!icn. A corresponding s:_cy o- -air:es_ ::.._. ._ s:_~ ; - _'css. o.._ •oL: w•i:: ge: an ...Cerirr. repor. C : =-c,;et_ of this year. WitY besc,personal regards, I a-: Very yo•,:rs, .:51~_f. -'_ ?eter _;:~er:.Se:d, Pi:.D. D'.r e:... r o: _'.esearc.. FL:?.:_ Eac:osurc IWD[PLNDGNT-N, ON•PPJF(; OPCRATCD ERC_USIVCL'• FOR Sc1ENTlF:c •tiD E7U=.TI_-V._ P_FP~;SCi CTR CaHTRRCT cz) 005325 11225066 CTR NN 042 351

3io-Resea-ch L^stitute, Lnc~roc:ated 9 Corr--nercial Aver.ue Cambridge, hfassacn_se::s Q' :=: ~ r'- Fee JUfV8 19'3 ; Y 1~3J ~J ZrJ Y J ~zJ ,~" Fir.al Repor: o.-. Contract CTR-70,'2 A Project E:t:t:ed xpanaed Stud-,• o: ',`_ouse Sb::.. ~ _..:e :_ . : o:e Fresh Smoke Cor'_ensa:_ Co^:pared to Sx:a :. T:=a _ ' ----- -.. - . . S. r.. Dated May, 19'? Prepared by Pe te r Be r:Se ld, P:.. D. 1.r CTR CONTRACTS 405~2~ 11225067 CT R M N 0 4 2` 3 'x2

i /k C•.:rre::: concez:s of :_-e biologic e.°fects of :oba:c~ smo:<e emanate from two principal sources, narr.ely (1) f:or.: in- vest:;ations on the statistical association of certain clinical man i- festa:ions in human populations with smoking, where cause and e:- fect can never be ascertained with absol;::e certain::.•, and (2) !:c: studies in vivo ar.d in vi:r~ on the ir-'luence of tobacco smoke cr smoke oroducts in exoerimental ar.imals. In the latter case, t-e ci:c-=-^'is:a::ces prevailing ir. the human smoker can n ever be sat:s:a:- tc:i:y sim•.:iateC, because of the differences in species anc i n the :.a tu-e c: the target tiss.:es, and because of the stress to t:ae ata=Ia:s res.a:i:._ from their .r.t•o:~n :ary exnosu:e .o toxic c: :ritant ::a._- ria:_. L^.ves:? ¢a:ions in :..e latter dom_ir. have bro•_,:: c:... one c: the most incri..•.r.atir.g and seemingly delete:ious r~ani.:=sz- t:_n= ca_sec by tnbaccc smoke, ..e. ...e aopea:ar.ce of c•.::cnec•_, pao?::o.:as a::= carcino: ias ::...._e as a co :seq:ae=ce of re:e:.:ec aco-::a::oae of =_: oke con_ensate :o t^e st:r of t:ese ar.i::za. The res_.ts of the prese..: wcr:< s:ron,iy sc¢_as: that w._ _a._•noFer.'c e°_iect o: tcbac_o smoke condensate .- -.._--_ s::'--. s ca•.:se-, at -ea_: to a la:R~ PL::, by ar. a-:icac:. ee~. ,.:.c-. .:es : ci¢a:e:ti s:r.cc... .-.•as __::de:.secd by .r:-.pLr, ..._n: C:- rrc::.. t: e s:<: :_. .::ice smoke a::= s'.a .•ican:;v feu•e- oaoil'.or:as and carcinor:a=_ ::e:e cause_ t- --ea-, _:=e!e ~._ ce :sate, ,...•' .:•a.^, . .:_ .._....':5._._:.. __. _..~\.~~. ..._ Gen e:a_ -=s:. n ..- ex:- r•..._r.. - ..._ _ .:...•. c: •..- expe^::::enta: •^'ia- is s'.z•x•n i.. _abie :. ::iice - I^ a;i, i, O1 : fe na;e C: F1 -- ~• ce;•:ec -rc:.^. T:e Jac:<scn Laboratory in Bar l:aroc:, "a::.e. C-n their arrival, t.`:e animals were from four to seven wc^ks of age. T: ey were distributed into six groups (Groups A throug:. F) in s.:ct ca- a rr.an ner that both the mean body weights and the bo'_y weight tri!;•.::ion in eac: of t:e six gro•,:ps were t::e same, as shok- in T a- ble 2. Animals were ho_sed in disposable p:astic boaes, covere,_2 with well-fit::cg, v.•i:e-screen tops. At t~ie begi:.r.in_ cf r* ..r ~ ~ CTf; CONTRRCTS 00530217 11225068 C T R I •N 0- 4 2 3 E5~"....

/k .:-e experir::ent, there ccere ten ::•:ice per cage in Gro--:)s A, 5 and C, a-.c either eight or six mice per cage in Grouos D, E an d F (see bclow•). Cage mates were selected so that there was the least pos- sible body weight difierer.tial amo-:g the a nimals in any one cage, in orcer to avoid do:aneering bebavior of large animals. In this x.ar.- ner, fighting among the mice was mi-•W ized, thereby prolonging their life- span. In addition, the inte rfe rence of wounds, inflicted bv fighting, with the appearance of skin lesions, caused by smoke ._=c.e nsates, was eliminated. As rr.ice died i.^n the course of t-._ ex=er:rnent, the n::mber of animals per cage was generally allowed :c decrease accordinglv. Only on two eccasions were rr.ice rea:•- :a nzed, -.: orde- to reduce t:he r.•.:r•:ber of cages, w:ereby animals, ..cich : ad :..,: been in the same cage at the beginning of the e:•peri- _. , were s•.:bsec,uentiy housed together. These two exceptions in Groups D, E and F after -. and 93 weeks of treatment, -•=s;eat those t:-iec, the -Ur-:e:s of ca;es irn eac: of tyes: _.ce gr:,cps vvere reduced f:rs: fro:n 1 to 12 (se_er. to eight rrice =z: cazej, a nd then fro~: :2 to e'.ght (five to eigh: rr..ce : er .-age' these , cDerat•..ns r_s__:e_ in _-ic.easec ~.crts'_i San-i-ce:, DeodDr served as ..._ be-_~c:n: rnatc- a_. C„cs and beddinc •.rere c: anvzd •:+eekly. The mi:e were kept :: Ous:. %•_..tila::d a nd :e::pe:_._re-_~ aed roo::: f-'c- "_ =).;•:-•) a:.tor::at:call;, cen.reLlec f:_cresce n. :bht (7:.. a. ••.. tc : o. T. ) wit: cu: any o::tside light. The anirr.ais ..:e:%•ec _ _r t::e p•_: pos_ of a'-: mice •xe-- . _ c. .=.:a'_-'. ca_- a:red at the beg?r :i: g- tc.e experi-ent acc :.a: ::zi_!. .a.t:,cec to r::ari. .:e No of G:o••. :.. _z.•_ C.;-as ._r•^ovec :r,,... ._._ an:...a: exce: a:ter sc _<- ..___s Zeatc cr sacrifice. _M:ce of r. E a: c. wer_ _ a...zr~ laboratory, four tirnes a just lon_ encug: to b^ ea_ :Se:. .. . ns:ant s:: ::<e ... :de :sate, „_._rwise ... were lCCD: ... :__ ar.:r 1 rco:r• in tc:e same r.:a.nner as the mice o_'Gro_ps A, E a~c C. Sha%;ng of mice - All an•r.sals, except a few r:-.ice kept as untreated and unshaved controls (not shown in Table I), were shaved by means of electric shavers on the'-ower parts of their backs. This was dcne immediately before the first applica- tien of test materia:s ar..'. periodical:y thereafter, usually twice a «•ee<, :. e. on Monday or "Wear.esday before treatment. Ln order 1 cTR coHTRRCTS 005326, 11225069 ~T R M N 0 4 2"" 3'53 -Ar

to prev_c: cor.ta.-ni:ation of the air wit: *-nouse hair, shaving :r_s carried out under well ventilated hoods. Application of the test solutions - Administration of precondensed smoke condensate (suspended in acetone), of lanoli= disso:c•ed in acetone, and of the vehicle alone (acetone) was r:ade fre^: disposable plastic 1m1 Tuberculin syringes without needles. The desired volume was delivered to each mouse by means o_' a Ea=ni::on repeating dispenser, which allows to mete out exactly 0. 0Z mi of liquid, or entire multiples of that quantity. Smoke condensates were ac.-:?r.istered to the an:-:.a:s un der a hood. All test materials were applied to rccently s;eve: mice on an approximatcly 0.6 to one-square inch area (7/8 to c,'S in ch in diameter) in the sacral region of the lower back, adjacea: to :he root c` the ._'?. Frecondensed s:noice condensa:e was prera:ed:=_... i:e nt-.:_:;: re:=re nce cigarettes by the b:elov iab=rat:r:es = Sprir.gfi_ld. t'irgiaia. This test material was shipped to cur ra- torics Lnder rc:rige:•aticn, was kcpt in a freezer and was use_ v.•::^in ..._• •;.•eeks of arr:c•a:. It ontai ned (•1~1v.) of r.c.^.aqueo_s s:r.o'Ke c,,..___sa:e, 4?"c w-a:er a nc 46°'c aceto :en A total cf :60 m¢ nonac,:e- o.:s condensate were, thus, applied per ^:_,:se per week, ad=in:s:er__ .. e~~al oorticns, each given on a different dav oi the w•ee:•:. __. _-pose of slow1% • hab::.:ating the ani:::als tc the ecu:e to:ci: _:fec:s con cai ::= i:, tne s:no•r.e condensate. „- one-_°o•_r:_ c-` :.. a.... ,.nt, :. e. =C mg, r:ere give-n du:ing the -•rst ueek, an.a tn .. - was gracnaay a-c regularly increased d•_r: :nc• subse .•.:e_: 1_ «=e: s the finai dose of 1oC mg was L^stant smoke con--'e=sate was a:sc , r eYared ~ar.._cc; re-eren ce cigarettes. A s:ighaie a.; :coria:e:: ...=c_..=_ -, .~_ reverse s^-:okiag machine (U. S. Pate n. No. -:?6, 11 use_ for t!:is p,.:rpose. Twelve cigarettes were smo•r.ec stn:u=-.e- o.:sl}• in an upright position, with the use of a ccnvenient:y a•?_ened (four inches) plastic piston (or cover) and an appropriate rn o:at:e base holding 12 cigarettes. One two-second puff was genera:e'_ from all 12 cigarettes every 60 seconds. The exposure chamber was re- placed by a small plastic mixing chamber of approzimately 6C ml volu:ne, leading to a ten-inch long plastic hose at the end of •.v'o:ch was a metallic orifice of 1/16 inch diameter. The latter was : e:= at a 1/4 inch distance f:om the mouse skin for smoke exposure. While -3- CTR CONTRRCT9 005329 11225070 r*7 CTR HN 042355

proc_c?ng the smoke, 3° m1 c: ai= were allowed to Dass t: rough each of the 12 cigarettes d,_rir.g every two-second puf.`, and when the 420 ml of smoke, thus generated, passed taro::g: the o-iSice, smoke condensate was deposited or, the mouse skin beneath. The amounts of in sta nt smoke conden sa:e, produced in t: is maaaer, and determined by alloacing the tar to condense on :i!xr oaper instead of mouse skin, were on the average 157.4 r-:g tar f:e-: 1.', cigarettes, or 13. 1 mg tar per cigarette. The amounts c: smoke condensate resulting from this manioulation were verified seve-a: tir:es in t: e ce•.:rse o_° the study. Taking into account tnat 13. 1^:g of tar were g°r.erated from r.i:.e outfs of ea: : cigaret:e, _:n:•ked to a ~...t lengt: of 30 it follcws that our pro:edure y:elds 431~, of the total amount of tar }zziowrr to be delivered from a 1: 1 F:en:tac}-; reference cigarette, as determined bv standard =asure::.e-a (J. F. 3enner, Proc. Tobacco and I:ealti: Con- -ernnce, iexingtcn, KentUck}•, Fe_.;:a-y, 1970, p. 3G). T'r.t ..•a:er cor.ten: of t.as ...stan : smoke ccnder.sa:=, as ~.ea=ured with Kar_ : isc^er reage.. , was 4. 65~ of the totr.'_ %ti•eight o: the .._densa:e, as op_osed :: 7.417, for :he cold-t-a= o:econde ns-z= smoke condensate. Care was :a•r.en ea:.-:::o;:se, trea:ec ...... ;--- star.: s^:oke condensate, receive--' (a; the same cuanti:y o_ sr.o:<e can '= nsa:_ _a._- day, as any ot:.e: ..n the grouP and (b) ccnden satc cc-. •:d _- _... an ccca: -.•.::nbc- c_' t__.. s: ccr.c, et-, , •::: (:::oke :rc... .: _ i:.-- was rejected) c.. - a perioc of _ne ..eek. :;.is was ac:ie•.•ed ir. At eac•rh sessi~r., ::.o%~se sa:e_ of p; :s c-'smoke from 12 ._;a-e:tes, Ibe;_ were e:.:e- tl~~: second and -....:h p•..ffs, :he third puifs, .^e four_n a nd se•:en ti: puffs, or the :ift : an d s:xth pu:_~s, as see n in Ta~1e L' .-. na= been established in preli:ainarv ea: eriments that eac:: cf t-ese four two-puff co:nbinatic.is fro^n 12 cigarettes always yielded ap- proximately eq•-ai quantities of smoke cc.^.densate, ranging from 35 to40rng. For practical p;:rposes, two smok:ng r.^.ac::°.nes, ooerated si:nul:aneous:y by two perscr.s, were used. L' each srroic- ing machine, two sets of 12 cigaret:es were smoked alternately at cTR coNTRRCTS 11225071 CTR ~'~h~~ 042356

30-se=enz :ntervals, so that the time intervals between twc p, `a re^:ain ed 60 seconds :or each cigarette in all instances. Since instant s:noke condensate was administered to the test a niznais c:z four days of each week, perfect rar.domization of the smoke con- densate application was accon:plished by employing a rotating se- cuence c_` exposure to the four di•'fere.^.t two-puff ccr nbinatior.s on each of the four days of the week. The sequence shown in Tab:e r.I is t:at used on the first day of each week. Or. the second trea:- *-r:en : day of the sa:r.e week, t^.e secuence irn which t~le four cages ca=e up fo= sr:•toke exposure, was shifted from 1, 2, 3, 4(fe: puSfs 2, 4 ar.c 5, respective2y, see figures in parentheses) t_ 2, 3, ,:. Oz tr_ third day c_° the week, the see,uence of cages was changed to 3 , 1, 2; oa the fourth and last treat^;en t day cf the week, it was fu:t :er modified to 4, 1, Z3;.n:il it reverted :a t: e original sequence of 1, 2, 3, = on the f:rs: day of the fo1::,c::=; J: .. In t:•is -:o 254' -..":e cou:_ be t=ea:.._ .:.: -.atan- s:r.cke ccnder.sate ir. o.^.E da ' or 4S0 r::_e _culo be trea:ea ic•,::• :..-es a«•eek on f:ve weef_ay s. I: was ad•ra nta¢eous to l:c-:se t: e a:.::::a:s nur•.:be: s of eit: er s:x or eight ~er :age, ac_ to =z:e er..-c r.-?::=--es =-- =ages .a eaci: Fro•_;. Fo. :cis --_- .--.:a! r.•_...c--5 of rnice :c- caz•.. Grc•_--_< D, _ an= _!se_ were _ig3: (i : eight cages; and six (ir. eEg^t caces). „ages oc;th sa r.:ice were used, e n'.y cne sToktng rr_aci:ine was neede: 'c° seco:._ rc __ ...ne - :s li.. ..._ _7:rer ...ne -_::es of .J.a_ ..- :ns:ar.t s7:".crce _..:.densa_ _ :ec cn "--_ bac.. .,: ea:'r. ...:-_- =. week was Ln c=ce: t: t;:ate 26 n.:::a;s :Z~' tze .c..._ _Le=-_ ;. 5...~-.r :...;..: _ i. t-.e sTc'[° ccnden9a:e. _n!,' -n 7:'^€ 7C instant _:..?~. u•ere ac-..:a:s:,!re.- to ea:: an.n:a_ the firs:_.._ ~.._ -an.. was s:_•x•: and increased by c:..__..-.. ...~r_ ;r.c na d;._.zg t: c f:rs: 14 weeks of .reat: :en: .., exa_tly the =a--." :ra=e: as for the preconder.sed s:roi<e con densate, unt'.1 t: e a no~a: of 160 mg condensate per mouse per week was reacn e_ d_-- ing tye fifteenth week; this level was main tained for t:e re:::a:nder of t::e exoeriment. Lanolin - A 5°•0 (w/v) solution in acetone was From C. C8 to 0. 32 r:- of that solutior. were adr.:inistered per mcuse per week, b;~ a sc^edu:e of gradually increasin g dosages the first 15 weeks, which is identical w';G•^. that follcwed for the azo_:can tion of the suspension of pre:ondeased smoke condensate in Gro_? A. -5- cTR caNTRRCTS1 oa"50333 T 11225072 CTR NN 0423SEr

L-. Crc•.:n E the lanolirn t:eatr.:ent preceded t:e apolicaticn :.: ia- s:ar.t sr: c:<e condensate by less than 30 ^:... _tes. Vehicle (Acetone) - Frc:-i 0. 03 to C. 32 r::: of ace- tone, t:^.e vehicle of the cold-trap-precondensea sr.noke condensa:e, u•ere ad...! :istered per racuse per week, by a schedule of gradcaliy increas;ng dosages d•.;ring the first 15 weeks of the expe::mect. This sc-eduie was identical wit:. those followed for the applicat:orn of prcc-~r._c :sed smoke condensate s::spension ir. Group:_ and of lar.cLr. sc:,:tion in Groups B and E. L-. Group F, the ad.:.inist:s- tion of ace:ore followed the treatment .:•ith instant s:noke con_en- sa:e W::::n ~= rnin::tes. Body we:ghts of mice - After weighin g ail anir~.-a:s incividualiy at t^.e tirr.e of distributing then^= into six gro•_Ds at t:e De3i:_ in_ ._ -..e exC)eri_-nen:, collective wetghts of aa a.:-::als .z- _~;:i:._ ..._ s;.e cage were determ'ned at r: onth'_ inte:•:als. ..z . ,::sible papi'o-:as were r.et included _.n th~_e ce_~c- .:ve w•e:g :.ngs, and :het: Nc•e'.g!as were de:er::iined separa:elv. From ::zese data, mont:.ly average bod;: weights were co^:pu:e= a_. cf each gro•_p, with a:.d v.•it'--out ;:ap::lo:::a<_. ~e?:nitior. e: = aa~il:e-a bt t:e c-:-er~.a e: ;ress -soec :o: 5ki.. ...egL:ar:ties c: less thar. 1.._.. ..n c!a-:e::-, ctser•re--~ on •r:ee::ly :.s pections were aisregarded. Lesions o: . rnrr. c- :-:cr~, . _: c: less ...an 2 .-.-.. in dia_:.ete: •4.•ere recor_ec as :ab:_ :lc as, , ro•:ided .r.a: t.ese lesicns s•_:ia:e -: t:e s::'.:. a.he^:. Les:c.^.s o: _.._.. o. ..._... be estabiiS'_.. Ja~illo:t:as; ....:e•_ , c...= _ esion =a_. reached or exceede= a d:ar.:et:r c_ 2 thc• t:^:c c- .._ rea_ -c ~ _.rst acoea:ance Ras ass.:^:e_ to be :aa: %c: en it :.ad firs: !.-__n c:a:ne:e:. Once a ies:on had bee_ accep:e_ as an esta'--:._; __ alc-a, its host con::nued to be ccnsidered as a rtiouee a _:c^. , e er. i: the lesior. reg-essea s_Sseque:,t'-y'.. L.c_ ~._c or «as sacri:iced. Mice witc: established papilioTas we:e sac:. -easor : _, :iced fcr _.:stologic studies for on e of the foJoa•ing three if t^e lesion had reached a diameter of o::e-ha.L inch, or (b) i.` the :_ sion had not regressed during the six months following its first dete:- tion, even though it had not reached a one-haL" inch diameter, or (c; if t:ere was reason to believe fro::•1 the general appeara nce that a:e lesion had become ^.zalignant. Multiple papilior.-.as ccc-u:red not infreq•-~enay. :... CTf~ CaNTf~RCTS 0Q~~~~ 11225073. V rl u/ 1 . 1  1 I 046v 3sV

a /k account of thc n~:zbers of an i:nals so affected and of the nLmber: of lesior.s per anima: observed was made at termiratio n of the ex- periment. For an evalcation of the diiferences in biologic respcr.se between animals o: the different groups, no distinction was made, however, bet•wee: animais with single or multiple lesions. The "ef=ective number" of mice - In accordance •-rtit~h a sugses:ion made by Bing^am and Falk (E. Bi_gha.^.: and H. L. Falic, Src :. ::nvirc Health 19:778-783, 19:9), the results of t.e papa:or..a incidence are expressed in terms of the nur: bers o-' ---:=e with estabi:s :ec a3p:Lor.:a= in oercent of the fleffect:ve nu:nbers" c= anir:a:s in ex:s:cr.ce e: the same time, rat.':er than i:. oercen: o: the numbers of s-_-:: vor s. The "effective n ur.:zer" cf ar.imais is defined as t: e of survivors wit's.out papillcr••as az a given time, pl•.:s the :o:a: r.u:::oer of animals with establis: ed papillomas, dead cr z.1:•:e, a. ::e sa~e time. The use cf the not?or. of an "e:_'ec_ :ive nur--e:•" c: a-.ir5 s:s satisf:es the assu ptica that t:e c_... _- of :..--= a:a, reac:: at any ,.:^.e, =_t may never e..- ceec _ nc _G ; ..._.... i~g:c studies - An?*:.a'-s u•::h papi:ioma_, e:-- __c. 1'.c r c:.e of the three reaso ns g:ven above, - ~e-`ore ter::..:at•::. :• .-e _.x:_r. ..._..: anc we: = s•_:_`wel: preser -vec, •.vere kep :.:. ne :ra_ :o: alceh~ d: _._ :.: _ pz:nologic sr-d:es of t=e treated area c: ._--...._.?on, ..c. after .0: .:•:e::s c: s:.... «•-._ sa_..:iced. The _res:... exciseZ s:.... -. ...- ..e a:cc a.ea _- -__ : _-. ra: was st-etc: eZ . :e- a s-.a1: -:e bca-d and __-c<= -..-..ec.atelv ir.:o Garno•;1> sc:_tio n. r-.1: skir. __.•s es were .ak.e:% c_. c. _::s fixative after ic r:....`s, curir.o ..:..c: _.r.:e tiss::cs :ac ____::e se:^itranspare-.:. Sic:- lesior.s . __c be CetecteC a: _~ac1e _rOts whic : woG:C :a•:e c: ie~•N:se e5Z37e= at:enticn; histe_c,:c sect;enswere cut t:rough suc: soots cr any o-her lesions. Ln t:e absence of any visible les:ons, skin sez_ tions of treated areas were raade at random, and all skin spec:-:.ens were transferred : nto alcohol and were stained with hematozy'_:: an: eosi:. T:.is was also carried out on the skin san:::es ke^: in fDr:na:- dehvde fro:a an:r.•a:s that had diec spo_ taneous'yo or that had bee-n sacri:iced earlier. Gon:o-ete a. :opsies were perfcrr.eZ r_ all ::ice -?- CTR CCHTRACTS 005252153 11225074 CTR HN 04~'3'vr9

a / h sac-i:iced on ter: :iaatic= _: t:^.e st•.d;: or ear:ier, as a•_i-i as .,•. ani-:zls t::at ha_ died spontaneously, Frovided t::at the :at:e- .ce: e su_'_'icientiy well preserved. 1?istoiegic sections were :~:ace o_ a:: grossly a!~no_-r::a'_ tissucs. _=.ES_LTS The i:.fo:rr.ation, forthcoming _'ro: : a study c: s:= -)::;- ccn_°r.sa:e application to the skin of experi.-ne :'-al zr.i :zts, scc:. a_ the _ rese-*_ invest:£at:on, can basically be d:v:ded into _°o_r czte- gories. _:ese are (i) acute and chronic toxic r.:an::esczaons, __ .-_.::ar r c-tal: y and char.ges in body weight o: t.e tes: aca.:.z:: ; (2) aoservat:crn of gress:y visible s<:r lesions i n the anir::a:s, .r. c:uci^; a.^n e:-al'.:aaor of both the extent and the rate of appeara:.., of s•.:ch :-sicns; (?) '~istc~a:^c:o;ic stvdies on s-..: sec:ions f-.,..• ce t_eate= arez c: the independently of t:e ._..-.- .n g: syster.::t effec:s. zs de:ected by gross .-- a_. ^s; his:a?a:ho:ogic examina:-.,- of ..._ toxiei:v e: ice - Co::.der. _ the .:..~ _ , r...._.. , ...~ ..-=-~1: , c_` ...• -..ice a:._. : :ox_c::: _: tr.e test :::a:e-ials, :•eze '-ca- in all six grc : _ __ee _ _- _.._ :_.._ _; ,%'-.-. a fe.r r•-:i::c7 ex.ept'._^s, ..._-_ %'--e-- n_ s: =etweer. :o•_- gr:-= _......__: - .- ---___ .,.t'r. A, L, E zn_ : ; ~cc .._._ c. %e•it: cold-.-ac-?recr:-nse= -..•c.._ __nsa:e; e:.:...,..__ z.- on e time a sc-r.e•x•hat !a;-`.e: ..._..,_... :.: s__- t: e.:: ee ...::e:• groups, i. e. Set•.vee : t::e ..~a ~... a..= _.- :_.._:. .c=_.e c_ t:ea:.-._.. . That timr pr:iod :. _.._..- ,,- t: e-a~--a:u^: a: cerr:aaent dose lev-_ ci _es: ...__. : appears 1 te:y, t'r.e-efo:e, t:at the ele•.a:e= ...:..z'.:`. __ :_ .-.-:.e - n Crcuo A at that moment was caused by an a:•::e .,...- e`_ec: o-` r.icoti-e in the orecondensed smoke condensate. -c obo,:s az;ae tcx::i:y was observed Ln any of the other grot:ps. An:m:.ls in the two control groups (Groups B a:'_ C; showed a somewhat lower mortality, than those in the ot=e- A s•.:~cen drop in survivors :r. Group B(controi gro-.p, treated •aa:•n lanola only) at 22 weeks was the result of accidentzl conditions and was •__re'_ated tc :.._ trea::ner.: with tes: materia:. I: thus a? pears CTR coNTRACTS 005334' 11225075 {.wI' I R /MI1 I 04236,0