Tobacco Documents Online

ANDREWS 0. YiCE PRODUCTS CAPITOL HEIGHTS, MD lK/ i

1980 REPORT oJ THE COUNCIL FOR TOBACCO RE3EARCII-U.3.A., Ire. TRE COUNCIL F()R TI11tA/'f:O ItF.Ct:Altfil-I/.c.A., lne. 110 F:.N 591h 11ree1, New Y..rk, N.Y. 10022

SCIENTIFIC ADVISORY BOARD to The Council lor Tobacco Research-U.S. A., Inc. as ot December 31. 1980 SHELDON C. SOMMFRS, M D., CA.Mwrw Dlnector of Loho•ato.kr. Leaos 102 Ila.pitai Cf/nkd ho%ssor o/ taAofory Co1k~e oI Phrsicians ! Surgear d CohnnDia Univenity New Yatt, New Yort RICHARD 1. B1NO. M.D. Dhedar of Cardiolngy and fntnrwa.l Mrdkier Hantington Memorial Ho.pitai, Pasadena, California Pbolessoi of Medieine Unhersitr of Southern Caiitornia School of Medicine Let Angeks, Cali(ornia ROSWELL K. BOUTWELL. Px.D. Peo%ssae of t7nrubtsy McArdk Laboratory lor Cancer Research thniversky of Wisconsin Madison; Wisconsin JOSEPH D. FELDMAN. M.D. Head, DepaHmettt of Immm"thobn Sa4" (lWe and Research Fotn+dation L.a loila. CalKornia Wi1.UAM U. GARDNER. Ptt.D. Sdentl/k DJrecao.. The Conncil for Tobacco Research-U. S.A., lax. Q. K. Haart Professor of Anetorny (enreritw) Yale Udvenit' School of Mcdicine New Have., C.rrrectkut ROBERT 1. tiUEBNER. M D. Chief. I aborstory of RNA Tumor Viruses National Cancer Institute Bethesda. Maryland i.EON O. IACOBSON, M D. loseph ReRenstein Pro/essor of Biolnairof Scienres i )niversity of Chicago I HENRY T. i.YNCH, M.D. rro%sso, and Cbee.ewn Department of Preventive Medicine and Public lleaith Creighton University School of Mcdicinc Omaha. Nebraska HANS MEIER. D.V.M.. Dr. Med. Vet., M.R.S.H. Senior Stel Sekwt/st The Jackson L.boratory Bar Harbor, Maine GORDON tt. SATO. Ptt.D. Pro/rssor of AtoWgy tlniversity o/ Calilornia, San Diego 1.8 Jolla. California Se1..liie $tat.f Tit. Cr.we/i WILUAM U. OARDNER, Ptt.D. Scfenft/k Dbscto. ROdERTC. HOCKE'fT, Pw.D. ReserneM Wrrclo. DONALD H. FORD. Ptt.D. V1NCL'M F. LISANTI, D.MD. Anociare ResreoswA iNrerctot " Asisdrtt Rssu.Y# Db.efer DAVID STONB, PM.D. Associare Resercb Direna.

CONTF.NTS Introduction Abstracts of Reports . . . . . . . . . . . . . . . . 7 Cancer -Related Studies . . . . . . . . . . . . . 7 The Respiratory System . . . . . . . . . . . . 29 lk.rt and Circulation . . . . . . . . . . . . . 41 Neuropharmacology tnd Pltpiologr . . . . . . . . . 49 Pharmacology .nd Biochemistrr . . . . . . . . . . 57 Immunology and Adaptive Mechanisnu . . . . . . . 69 Fpidemiology . . . . . . . . . . . . . . . . EO Active Projects . ... . . . . . . . . . . . . . . . 90 Completed Projects . . . . . . . . . . . . . . . . 99 Inde: o( Pri.cip.i lavestigaton . . . . . . . . . . . . 111 Inde: of Senior Authors . . . . . . . . . . . . . . 112 Int: oduction Since iis formatioe 27 years ago, the Scientific Advisory Board so The Council fw Tobacco Research•U.S.A., inc., ha reviewed many formai awd i.- /ormal applications for research support. The direct resnk is that 413 diAereM projects have been /uwded by The Council. based upon the reeanmendNios of the Advisory tloard, 1o supporl investiptors in 2S9 medical schooh, hospwah .nd research institutrons. Each ol these projMs, whether active or eoanpleled, is listed in Ihis year's Report. Total fwwlinR through 1900 was f61.00(1.000. The chief output of The Cornci/'s tln.wei.l support a.e 1.862 scieMiOe papers and reponsl published by drese invesliwors in rwany diAerenl medical and scientillc journ.b, covering a wide range of srbjeets. The publicatiow of these research resnNs .dds to the sciaNilk literature a.d to the stoehpik of information from which, it is hoped. will come .wswen so cornpks proMewr such as those associated with uncer, cardiovascular diseases and puinrow.ry ailments. Abstracts of wunuscripls published during 1980 that actnowledRed Council support comprise a sectiow of tbi. Mww.l Reporl. During 19tl0 two new wremhen were added to the ScieMifie Advisory Board to provide additional eapertise to that distinguished body. They were Roswelf K. Roarwell. Ph.D.. Professor of Oncology at Ihe MeA.dk Laboratory for Cancer Research, University of Wiseorrin. Madison, and Gordon H. S.Io, Ph.D., Prolessor of Biology. Universily of California. San Diego. I 5

Abstracts og Reports I Following are aMltacls, approved by the .ulhors, of reports on .ew research aclnowkdtint support Irom The Council that have appeared in scirw IiBc pwrnsls since publicalion of the 1979 Report. The name of 1he recipient is in italics. lhe ahuracls are Rrouped under these headings: 1. Cancer-Relaled Sludia, 1'. The Respiratory System. Ill. Heart and ('ireulalion. IV. Neuropharmacobpr and Phrsiologr. V. Pharmscobpr and Siochemisuy, VI. Immunolbay and Adaptive Mechanisms, VII. Fpidemiology. 1. C.wcor-Rot.t.d Stssdl.. ARYI. IIYDROCARSON HYDROXYLASE IN NORMAL AND CANCER POPULATIONS The roks of aryl hydrocarhon hydrosylne (AHII) and its inducibility in polpcyclic aromatic hydrocareon earciwoRewesis ue discussed at length in what is essentially a review of the research iw this .re.. With respect b Iwww AIIII, /his poup's cwrenl .iews are sunwn.rittd as follows: ( I/ the enrlnrte's inducibility diRen among variow Mdi.iduab; (2) this di/lere.p is pe.NicaNy con/rolh:d; ( 3) the distribution plterw of ANN Iwdnehy in the normal pnpolation may merely reAecl d.y-lo-day vari.Ailily In the lymphocyte arslewt, but it also suggests polygenic eoaMrd of Mte enzyme system; snd (4) lrsy and orophsryn6eal cancer patients have a AiRlter A/IH inducibility tw Mtl found in /he normal population. Indications are. however, thal cancer per ar does um cause high ANII inJrcibilNy. In plients with other types e( wNR- nancies. the enzyme inducibility was indialiwRttishab/e from that found i. the mKmal population. Studies Ihm show aw iwcre.sed iwcide.ce of hydrocarbo.- induced wmors In animals .uscepible so ANN irduction, harever, stNResl that such inducibility i. a eriliral factor M•1A. oecnrrence of chewtic.llp W duced turnun. Never/hdess. Ihe deve/opwrcnl of a more rdiable melhod of . assessing AI/II br/ucihiliMr in hwmns is Orersyuisile to Ihe es/aMisMee.1 ef this factor's role as a determinant of wsceptibilitr 1o hydrocarbon-LAuoed lumors in man. Arnwr, M. S.. Yamuchi. T. and lohson. D. A. In: Grifiin, A. C. and Shaw, C. R. (eds-): CwriwoKewsr /denti/k.rion .nd Mecbuniuns o/ Action. New Yorh! Raven Press; 1979, pp. 113-136. Oth.r swppert: National Institute of General Medical Sciences and Nalio.al Cancer Institute. From the Ikpartmcnls of flioW.ty and Sioma/hematics, The Univrrsily of Teass System Cancer ('enter, M 1). Andersoa /lospital and Tumor Inailuu; and the Umvcrsity ol Fesas Mcdical School at Houston. 7

ARYI. FIYf)ROCARBON NY1)ROXYLASE IN IIUMAN CANCER POPULAIIONS lhe genetics of aryl hydrocarbon hydrosylase (AIIN) inducihility Rod tumor suscepribilify have been sludred eatensively in inbred mice, and some of the eaperrrnental animal models have sugjesled that AHII irMlucibilAy could be an important delerminanl of susceptibility so hydrocarbon-induced Cumors in humans. 10 lest this hypothesis. AHH activity was measured in cultured human lymphocytes. One rnvestrelNor, swveyinR inducibility in a poprlaliun of 35) healthy donors, repwted that it (eN into three groups: k+w, intermedi- ate and high. lhn trimodal distributio. suggested that two slkks at a sin6k locus were responsible for the genetic control of AHN inducibility. Family uudies were consistent with this eowcepl. The present invesliptors measured AII/I inducibility in lung cancer patients swd palierws with other Iypes of cancer, snd developed a sophinicded, consppMerked syNem for information rettieval pertaining to normal dowors and ea.oer palienu. Over 700 normal d.+nots and 600 cancer patrems, inc/udinR approaimalely 250 with lung can- cer, have been tested in this system to date. The studies reported here lead to the following summar7 of A11H induabilily in humans: (1) individuals difler in therr AIIII n+ducrbdry; (2) the inducibilily differences obsersed be- tween prople are under genetic comrol; ()) the rnimodal distribution o( AI1/1 inducdwlity observed in the normal population may stem in p.rt from day-to- dar variability in the lymphocyte syuem, bul aho suggests poyflenic cawrd of the enzyme sysuem; and (4) lung and oropharyngeal cancer patients ea- hrbrl hrgher A1111 rnducrAMhty than the normal population. No cause and eReet relationship can he rnierred from these studres alone, but viewed in the contesl of what has been shcwn clearly in e.Perimenul anmmsls, the human lymphocyte A1111 uudret suMe.t that AFl/l rnducrhdd7 rs an imlrrrlant de- lermmanl of suscepahday to hydrocarbon induced tumors Arnorr. M S. Yamauchr, T and /ohnsron, 1) A /n: /ones, P. W. and 1 eber, P(eds 1: Polrnrt lear Aromatic HydrorarAont, Ann Arbor Science Publishers. Inc , 1979, pp. 779 791. Other s.rp.rlt National Inslitule of General Medical Sciences and National Cancer Institute. From the Departments of Biology and Biomathemalics, The University of Teaas System Cancer Center, M.D. Anderson Hospital and Tumor InuNute; and The University of Tesas 1leshh Science Cenler, Medical School a1 Houston. ARYI. HYDROCARBON IIYDROXYLASE ACTIVITY IN PUI.M()NARY ALVEOLAR MACROPNAO!'S AND LYMPHOCYTFS FROM I.UN(3 CAN('ER AND NONCANCER PATIEN7S: A CORRELA'I ION WITII FAMILY IIISTORIES OF CANCER In this attempt to determine whether differences in aryl hydrocarbon hydrosylase (A/lll) irducrbdrty esist between normal and lung cancer pa- uenrs. A1111 actuvNy was maasured in pulmonary alveolar n.actoPhages (PAMt) and prrrplwral hio.rt ItimpMrcyies from cigarette smoken with and without primary lung cancer. The dals allowed comparisons between 1ht nor- rnal and lung cancer patients for 26 variables, including sile, occupalion, aes, and family histoties of disease. Frequency dislribtMion analysis of AHH i.- duclion ratios for the two groups revealed as increased aumber of iwdividtals in the lung cancer patient liroup with high lymphocyte induetiow values. Sratil an increase was not shown fot hi"AM AHH values in hrsg cancer pslie.b. When individual PAM and lymphocyte AHN values wen compared belwee" noocancer and lung cancer patierws, there was a positive eorrdslion for now- cancer patiems, but /hese values did no1 correlate (or hswg eaacer paliewu. These cancer paiieMS were divided into three subgroups dsowinR (•1) Msli PAM and low lymphocyte AIIII kvels, (/11 low PAM awd low lymphocpte AIIII kvels, and (111) low PAM ard high ymphoeyu AHH kvels. Whew the incidence of family history of eawcer was compared (or lhne•subporps, none was recorded /or persons in srARroup 11; however, individuals in sub- groups I and 111 presented family cancer history iwcideaces of 9.5% and )9.1%, respectively. Patients in group 1/1 averaged sia years younger Ihr those in group 1. These dats support the hypothesis that high levels o/ AI/N might be associated with increased levels of hrwR cancer In rnat, and Ihey indicate that, for the sn.ysis of A1111 levels in hrwg eancer palients. Yse of the cultured lymphocyte system as a sole indicator of high All/l levels ia .o1 warranted. These data sho indkale that genetic facton are espressed In the initiation of lung csacer. Mct.enwre. T. 1.., Alarrin, R. R.. Springer. R. R., Wrsy, N.. CantreR, fd. T., srd Susbee, D. L. Rloc/iemk./ Cenerks 17(9/ 10):79S-fi0S, 1979. Otii.r auppor/: National InWilules of Health, National Cancer Irrtitr/e, Americsw Cancer Sociely, awd /he Veleraru Aderinistraliow Hospital. HorrNO.. From the Department of Mediciwe, faaplor College of Medicine. How1o.; Veterans Administration Hospital. Houalon; Deparlnsewr of Pharrnacolop, Teaas ('olkge of Osteopsthie Medicine. Fort Worth; awd rhe Deparlnse.l d Biological Sciences and the Genetics CesNer, Nor1h Teaas S1a1e University. Deelon. I DOSE-RF_SPONSE RELATIONSHIP OF RAT ARYI- HYDROCARfaON IIYDROXYI.ASE AND EPOXIDE HYDRATASE INDUCTION The data reported here sapport 1he hrpolhesii thN different regalatory mechanisms control sryl hydrocarbon hydrosylase ( All H ) and eposide hydrNw (EN) activity and that h selective inducer o/ liver entyates, auch as pheno- barbilal, may also affect the inducibility of estraAepalie monooarflews.ea by other chemicals. The Allll in ra1 h" and tidney is highly sewsilive so I.- ducers in cigarette snake and smole cordcraale, wfsile EH activity is na1 a/- (ected by these agents. Both AINI and EN can be indreed selectively is pri- rwary rat liver cells in culune. I.ow doses of beaw(.Isnlhracene (SA) prefer- entially enhance Allll aclivNy, while rrans-sliRsene oaide (ISO) and vuiorrs e 9

.~..1. .~f . . anriosidants affect only the F11 activity. Phenobarbital, which potentiales in- duction of lung AIIH activity by low doses of benro(.)pyrene, also induces AI111 activity in cell culture and has a greater than additive effect when mised into the culture nxdoum with BA. The effect of phenobarbital on A1111 induc- lion is M.K\cd by TSO but not by the ioducing effect of BA. These results suggest that phenobarbital is not merely an A1111 inducer, but also has the capacity to potentiate the action of cheereals that belong to a direrent class of smhrcen. In parlicrrlar, they derrwnstrMe that phenobarbitsl a/one induces  cylochrorne P-ISO linked A1111 good causes the accumulalion of rnore cyto- chrome P,-450 associated enzyme whew rrrised with BA. CirGn, 1. E. rr .1. ArrAivr, o/ Torknloty Suppl. ):20l-216, 1980. From the I.aboratoire de Chimie MEdkak, Instilut de Pa/hologic. Univenilf de I rtje, I ii`le. Belgium. COMPARItON OF ARYI HYDROCARBON HYDROXYI.ASE AND FPOX11)E IIYDRATASE INI)UC"IION IN PRIMARY FETAL RAT LIVER CI't.t. ('UI TURE The effects of benz/alanthraceae (BA). phenobarbital (PB). cigarette smoke condensate (('S('i, 2.).7,t.tetrachlorodibenzo-pdiosin (1('DD) and rr.nr-stilbene oside (TSO) on aryl hydrocarbon hydrosylase (Alllli and eposide hydralase ( Elf ) activity have been studied in primary /etsl rat liver cell culture. The two enzymes vary iw their response to lhese chemicals as shown by the following: (1) the concentration of BA. PB and CSC required /o induce Allll is lower than the one treedcd for signi/kanl irrduction o/ El/; (2) TCDD and low concentrations of A aekctively induce AHII; (3) with BA. PB and CSC. the ANN induction kinetics are not the same ns those of EH itduction; (1) TSO ieduces EN selectively. In the early phases of EH induction, RNA and protein synthesis r well as the eontimrous presence of the ir.ducer are required. When EN activity reaches a plateau, neither iaact RNA uor protein synthesis is required 1o tnaiMaiw enzyme activity at its opti- mal level. The Ell acti.ity decays with the removal of the inducer, and its biololic haN-Hle Is eslimaled to be abow 72 hours as opposed 1o about 10 horrrs for A1111. While TSO prevents the irduction of A1111 by PB, is does nol block that twcdialed by BA and CSC; but added together wah PB, BA, CSC or PB plus BA, it Induces the F.H activity in a more than additive man- t.er. TUis eflect, however, is only seen after sia days of continuous Ireatmcm. According to thesc results, in this particular timre eultute model, the mech- anism of Alll/ and 1'.1/ induction can clearly be dissociated. (Ioujow, F M, V P-n Canllort, 1, and Cklrw, /. E. ('A....k.. N..J.•rr..1lwILw/N.nr 1? M1 175, 19RQ I r.•n. 11v 1.r....I ..... .k 1 h.r..r Mtdrak t~ lu.uulu~~e, In.l.w~ Jr Pa~hoN~~~e, (1n..e..,i1 .1. I 1 r. 1.1 p M./r.um i I CORRI;LATION OF INDUCIBII.ITY OF ARYI. RIYDROCARBON IIYI)ROXYt.ASI: WITH SUSCEPT1B11.ITY TO 7-METHYI.CIIOLAN- 111RENI:-tNDUCNb LUNG CANCERS Intratrached treatment with polycyclic aromatic hydrocarborts may iw- duce specific genetic regulation of AHH adi.ity. Previously tned methods for producieg lung carcinomas in inbred atrains of mice wuesl a possible aaimal model in which the susceptibility to pulmonary carcinomas may be specifieaNy linked to the capacity of that organ to wrelabolite chemical earcinoRens. Iw Ihe system described here, mice of the CS7BIJ6Cum. DBA/2Cum, firsl flli. (F,)Snd bactcrots progeny from the two parental strains were evaluated /or susceptibility to )-nrethykholanthrene (MCA)-induced lung cancers. In /he crosscs, AI111 responsiveness segregated as a single wlowmal dominant aene (A/i locus) The AHII responsive mice esprened a very much higher A/IN activity/g wet weight liver alter intraperiloneal treatment wwh MCA tham the nonresponsive animals. /ntratraeheal administration ol M(-A (four 500 ang dores, a1 weekly intervals) caused a.ariely of pulmonary malignancies among the mict surviving one year after IreNtnent, inchding sauamous cell earci- nomas, alveolar adenocarcirwmas and adenosqwmous cell carcinomas. The A11H-responsive CS7BL/(iCum, F, and C37BL/6Cum a F, mice were wweh more susceppibk 1o MCA-induced lung eancen than the nonresponsive DBA/ 2(•ums. Furtherrrw.re, /he lung cancers found in DBA/?('wn a Fr bactcrors progeny did not occur randumly, since aipwOeantly more tumors were found in the AIIll responsive offspring rhan in the nonresponsive ones. Thesa data knd lo support the eaistence a/ a genetic link between the capacity to respond to polycyclic aromatic hydrocarbons through increased levels of AIIN activity and susceptibility to certaiw types oi maliRnaweie.. Indications are /hst aa- ceptibiliq to MCA-induced WnB earcinorwas is related 1o the AM allele. Ttieae results also suggest that animal modeb i.volviwg 0bred strairu could be trrd to study the control and repdMiow of epMhelidly detived tumora, tha lype moal /reaue.tly observed in the hurw.. Iwtg. Kour:, R. E. rr d. (Afkro6lolopkd Asrod.res, lnr.) Cancer Letters 9:277-2t1, 1980. Other w'poorfr National Cancer IratitWe. From the Departmewl of Biochemical Oncology and Department of Eapt:ri- mental Oncology. Microbiological MtiociMes, lr.e., Belhesda, Md. XI?NOIROPIC VIRUS EXPRESSION AND SIISCEPTIBII./TY TO )• Mt 111 Yl.(-11OB.AN I IIRE N!:-INDUCED CANCER Mice front two inbred urains. NZS/SI.NI and 129/1, nrd their variar genetic crosses were studied here lor evidence ol a genetic linlage betwass spon/anous production of in/ecri«ws aenwropie (X-tropic) virus aad auscsF tibility to chemically Induced cancers. The pantnlal alrawn and Fo, bactcror, and F, progeny bNween thcse two strains were partially spknectomized to M- oertain X-tropic viral status and were wbsequenNy treated .ubculantously, wwith SOtI r6 of ) melhykM.lanthrenc in tri.rctanoin. Progeny /rom second bact- erosses were also le.tcd lor Iheir X•Iropic viral status and suscepibility to l-nKthylcholanlhrcne carcum.gcnctis. All mice were obacrved (or evidence of 11

fibrosarcomas at the site of inoculatioa over a 10-momh period. In order to specifically address the question of she role of X-tropic virus esprcssion on susceplibility to chemical carcrnotenesis, populNiotss in which vwus espressNon sepega/es were suudsed llere, the carcinogenicity of she chemical was deter- mined by obtaining a single value, the C1, which uuilired both tunusr incidence and latency period. After )-methykhdantbrene treument, 42% of Ihe virus- positive mice developed tunsors r cowtpared to 22% of the virus-nepti.e annnals. Ilowever. the average latency period waa much longer for the virus- positive mice. so that the resuNaat CIY for Ibe two populations were quite similar. Overall, the resuMs seew here suggest that Ii1tk, if any, correlation esists between virus espression and 9-wntbykholanthrene earcino6enesis, and that there is no densonstrabk genetic linkage belween the Iwo. Nayar, K. T., Levy, /. A, O'NeiR, B., aa/ Kowl. R. E. (MicrobldoCk.l Asroci.res, Inc.) l'.ncer Reree.rA 40.1161-1)67, 1980. From (homet Technologies, luc., and tM Department of Siochemical (M- coto6y, Microbiological Associates, (ne., Oetheda, Md.; and Cancer Rcoearch Institute, Uaavenity of Cah/ornia School of Medkine, S.e Francisco. COMPARATIVE FFFE(-'TS OF INDOLE AND AMINOACETONITRII.E DERIVA7IVFS ON 1)IMETIIYI.NITROSAMINE-DEMETNYLASE AND ARYI. HYI)ROCARBON IIYDROXYLASE ACTIVITIES Certain substances such as the 7-substNuled indole derivalives found in crucrferous yegetabks, including bru..el spouts, cabbage and eaulsllower, alter the biological responses to chemical eucino6eos. Of these eompounds, which induce aryl hydrocarAors hydrotylase (AIiH) and inhibN polycyclic hydro- casbon•induced neoplasia, at knt one, )•indol7methanol, she induces amino- pyrine Ndcsnethylase and p-nilroaniaok Odemethylase. Indole itself also in- ducts dimethylnilrossmine (DMN)-&netbylass I in the rN, as does erypto- phan. Mnino.cetonitrik, a nsiaed-Hwtdion osidase modi/kr, iMerteres with DMN ntetabolisrn, inhibib DMN-induced rnethylation of Aepatic RNA and breakage of DNA, and protects a#aiwsl DMN-produced injury to micsosomal amino acid incorporation; it she protects against DMN-induced disorRani:a- tion of hepalk fine structure and DMN-i.duoed hepatic eareinosenesis. Here. she eRecss of indok and amino.utonitrik derivatives on the two enrymk forms of DMNdemethylase and on A/I/l were studied in viro. Indole, ) i.- dolymethand, 1-irdulylaceloniuile, )-indolylatetone and 1.•lrypksphan in- creased A11/1 induclion l- to 6 fold, but p•)-iwddyk(hned had no e/fect. In fact, Me latter deerea+ed she tissue endoplasmic reticufum contenl (/e., micro- somal protein/unit weight) by 21%. Only I:tryplophan induced DMPI-deme- thylase I and only it and )-indolymethsnol induced DMNdemethylase 11, about doubling the entyme activity in all three instances. Anwta.cetonitrik strongly repressed DMNdemethylaae 1. Substitution of the .rnino group greatly decreased or abolished mised•function osidase repressor activity. For esampk, imimdiacetonnrik displayed only ahout one•R1th the repressor ac- tivity of the parent cnmpiwnd, while nitrdolriaeetonitnk and dimethylamino- acetomude seemed to he inactive Ncither the derivatives studied twr <he par- ent comlwund, huwever, had any eflect on 1)MN-demethylase 11 or AtIH activities. Since these mised-function osidases represent critical steps in the rnetahulnm of DMN and polynuckar hydrocarbon., she modifying eRects of iodok and aminoacetonilrik and their deri.atives demonstrate she potential compksity of assessing she eRects of dietary constituents on the carcirakc.k responses. It is suggested that such constituents can generally aber ausceptrbilay to chemical carcinogens and in some innances, can further amplify the car- cinogenic responses to one or nare combination of agents. Arcos, !. C. rt d. C.ncer Leneri 9:161-167, 1990. Ot16.r a.'prt r IloRma.a-La Roche, Inc. From the Seamen's Memorial Research Laboratory, U. S. Public Health Service Ilospitd; and she Depa.ement of Medkine, Tula.e University Medical CeMer, New (Jrkans. 111(i11-PRESSURE LIQUID CHROMATOORAPHIC ANALYSIS OF flEN2O(a)PYRENE METABOLISM BY HUMAN LYMPHOCYTES FROM U()NORS OF DIFFI.RENT ARYL HYDOCARBON HY- DROXYLASE INDUCHlILITY AND ANTIPYRINE HALF-LIVES Lymphocrtes from sis hunun donors were evaluated by meaes of bi6h- pressure Ipud cbromatography (HPLC) for tbeir ability so wtetabolin bewo(.)pyrene (flP). In these subjects, tbe aryl ttydrocubow bfdroaylna (AIIH) irducibility ratio ranged from 2.4 to 1.6 and the aaipyrine plnrraa hdf-IJe from eight l0 17 bows. Whik the results show that f!P metabolizing activity does not diRer qralMMively amowR hwan dowors, there .re qtrrlMa- tive variations in the isducibility of SP wrctabolites, reReNi.s known individud differences in AIIH inducibility. Several fIP metabd'stes were iaeaiRcd: 7,f1- drhydrodiol, quinones, and 9-brdroar and 3-ttydrosy phenoh. The HPLC data for the induction of SP phenol prodw.yitM oenelaled well witA she kwwo A/ill inducibility ratios for she dowors, as delernsined by the tosveMiow.l fAwromttrk AIIIf assay. These stssdies she indicated that the induction of bnuo(.)pyrene-7,tdibydrodiol, proposed a/he proairaate carcinogenic foew of flP, does not parallel RP phenol indueliow. 1s addition, there was a s1ro.R negative correlation between AIIH inducibility and measuremeas of p/astw4 antipyrine or urinary •-bydroayantipyrine half-life. At best 1hen. AHII in- ducibilay can be presumed to be an indea of the capacity to nsetaboliru IP through nsajor, if no/ a8, patbwars, wRRestinR wseyual /lue of BP Mroup/s the various routes. The data also indicate thN at kan one activalin6 iner- nsedute systcm, the formation of 1!1'-7,rdiol, is not directly proportional to AI111 kvels. C aroo. H. L., Vau6ht, 1. 1l., Mdtinelks, A. /., Pai6en, 6, Ceisner, T.. ad 1lolanowska, W. C.nrer Research 10:1 T0S-1)10, 19r0 OtAer swrrort t U. S. Public lleahh Service. From the Deparament of li.perimental Therapeutics and Grace Cancer Drug ('enter, and the Ikpartmcnt of Molecular Biology. Roswell Park Menoorial Institute, BuRalo. 12 11

I ('AR('IN(NiliNlt'l l YO1:HI N/O(o)PYRI Nli ANI) IIIIR-11:EN (117 IfS 1)1 KIVA I IVI:S IN (')11/1('am MI<'f lhe tomorifeenicity ot a selected number of hento(,.)pyrcnc (HP) de- rivatives was inrestotaled in a tunwn model consntin6 of fiMusarcoma indoc- Iwn in suhcuNaneounly injrcted (')11/(Cum mice. lhe It derivalives used were selected on the basis rd their !n rbro and in rirn hialogie aelrvily, as well as o( their prNtnlial foH ehKnlalinK the .rnponance rd HI'-7,K-dwl 9,tlt.cl+oaide in mediating lumor lormati4.n Io1lo.inK subcutaneous HP administration. lhe resulls suRRest that mduction of subcu/aneous fihrosarcomas in thn mouse strain has omportant limitations in spite of SP's ability to cause a high inci- dence of tumnrs. Most signdkantly, she derivalives' polarity appests to mark- edly ahcr Ihew hMAopie aahvity in /hia system Because nrelabolites of poly- eyclic hyJrocarhons are usu./ly more polar than their parent compound, il is ddlicuh to assign a particular one so the a1Nus of proaimate or ultimte car• einnten In additoon, wnh any given eompound, the lurnw 'incidcnce was notably aflecled hy the nature of the solvent. For eaampk, SP's carcirw+teni- cny index decreased about S0•70% when DMSO was used as she vehick rather than trK+ctanoin. (M she other hand, other investilators have found 1h.1 It was almost completely abolished in a/:1 Irioctanoin and beeswax mixture. According Io some previnw reports, tumorigenicity of BP•7,t-diol-9,10- eponide dustereomers on mouse skin also depends on the soWent employed, the highest incidence having been observed with teeltahydrofuran and wilh acetune. Both of these, huwever, proved to he toaic to mice. AddAional re- search may lead to the discovery of a solvent or solvent combination that mini• mirn she efleet of a compound's polarity in determining tumor response. ' Kouri, R- E. .r ./ ( Mir.oAwfotird Assari.rrr, Inc.) 1or.nd o/ the Narioe.J C.nre. lnrrlrrrr 61( )):617-62 ), 1990. From the Depntment of Srochemiwry and Drug MetaAolism, Ilof(mann-l.a- Roche, Inc., Nuwky, N 1; the Department of diochemical Oncology. Micro- biological Associates, Inc.; and the Section of Oxidation Mechanisms. l.aboralory of Ilioor6anic Chemislry, National Institute of Arthritis. Melabolrsm, and 1)iKes- tive Diseases, Oelhesda, Md. SI'CONDARY M1: f ASOLISM OF SENZO(a)PYRI:NE IN I/UMAN CEI.IS While manr mNabotites of benro(.)pyrcne (SP) are known to form in man, receM s/udies have shown thal /hese primary metaholiles are /ransformed further to conjugates of sutfale, glucuronide and gluthalhione. llere, human tymphocytes and lung macrophales were used to study DP metabolism in cancer and noncancer patients. Results oI Ihesc esperiments showed a reeta- Mdoc pro8k similar to that reported by others for human lymphocytes. After hydrolysis of the fractions, the metabolic profiks were slightly altered. The observed data suggest that during a short lime of reaction whese the substrate concentrat«sn is not saturating the enryme compka, a substantial nmown/ of conlugNion occurs via Klucuronide formation lhe phenoh are conjugated to an equal estent wnh glucuronKk and sulfate. Also, in Wng macrophatees the 14 I estent of cunluKation was variable from per.on to person A preliminary aw- vcy u1 livc nom.m.skcrs indKalcd that the extent and selectivity of ineubol- ism was variahlc as well. lhcre was no consistent pattern Io the varialiow in conjugation ol 01' reetahohtes. Overall, the data presented in Ihis paper i.di- cale that cultured human lymphoeytes and pulmonary alveolar teaeropAap have the capacity to metabolire S/' Io several hydroxy and quinone derivatives. lhese arc lurther metabolized 1u polar conjugates, including sulfate and glu- curanwks 1 he proportions of the vanous pimary, seelabolites and eonjuples vary between individuals and vary with the tirne of incub.tioa and eP concta- Iration in a single individual. IMn increasinp and compoundinR the complexity of BP metaM.lism in human tissun. C.ntrerr, E. T'. .r J. rro.reJinRr o/ rh. Wruirn Phwrn.roluey Socirry 22:27)•276, 1979. O/h.r aupp.rlr U. S. Public /leahh Service. From the North leaas Sta1e University /ltahh Sciences Center; Colkge of Osleopathic Mrdicine, Fwt Worth. lea.; and M.D. Anderson Hospital and Tunwr Institute, tlouslon. eF.NZOIoIPYRENE ME1 ABOI.ISM IN RAT FETAL HEPATOCYTES (:UI.TURE. IMPROVED METIIO()OLOaY AND EFFECl' OF SU SSl R ATE CONCENT R Al ION lhe exact rate o( henzn(„byrene (uP) metalsdism in any living syMetn is imponant in that the balance betwcen tht r.riow enzymatic reactions b- volved in this process is highly signi/kanl in deltrmitwn4 the iMtacelWlar ten- centration of the toxic intermedi.tes. Presented Aere :re data on SP tectabolistn in primary letal rat liver cell culture. The tatenl of iw r/ro 1'H)SP eseuboNna was established by measuring all of the tANabolitn retained iw the aeM aa weR as those excreted into the cuMure tnediutw. lAt eaent of the eoajusaioe srd the nature of she conjugates were determined aed the nxtabdite paltcra .aa analyzed by high perlormance liqnid chromatography (HPLC). The fetal hepatacytcs actively metabolize SP and readily e.crele aR the metaboYtes iao the culture medium as wllates and tlucurunide eonjugales, lhe relative pro- pwtion rd the latter varies directly as,a /unetinn of she SP concentration. As shown by the IIPI (' analysis. eP•1,6-quinowe aad -),6-quiaswre are tlle mN.- boldes excreted in she larVit aaounl, suke.ting the probable existence of an active 6 hydra.ylase reactiorn medtanism in the fetal hcpatoeytts Aryl hydro• carlM.n hydroaylase activity nwddkti she overall rate of /!P metabolnm dra- matically but does not a/fecl she qualitative paltern ol Ihe excreted melabolites. Van Cantfurt, I.. (loujon. F. M. and (lirlea,l. E. CArrait'u-Rioluriralln/erurN..nr ?K:117•160• 1979. From the l.aborioaqrc dc ('himic Medreak, lnsutus de Pathobgie. Udversitt de I .ttKe. 1.i2Ke, /irloe/um. IS

I PHA(:O('YTOSIS OF ASS1:STOS FIBERS BY HUMAN PULMONARY ALVEOI-AR MACROPHAGES Ahhough epidemiologic evidence supports a relationship between esporure to asbestoa (AS) fibers and the incidence of cancer, the eaact nwthanism of '•aclion on the human respiratory tract remains unknown. The pulmonary alveolar macrophages (PAMs), which ingest various foreign pirticks that enter the destal tracheobronc/aal ttse, are also the primary line of defense a6ainst inhaled AS fibets, bu1 beeause it M virtually indestruaibk, (his material poses a unique probkm for PAM dnoaiReation. There is evidence, however, that pAa6ocytous followed by eoaling of the panicles with celi memb.ane componeMs, acid mucopdysaceharidn aad hemosiderin convert it to a less tosic form. The present study investipled aome biological iwueraclisns between cultured human PAMs and aewsite AS. Phallocytosis, cytolo.ic.ty and cell surface changes were esamiwed by acannin6 ekclron microscory a/ter the cetli viability had Rrs1 been delermired by the trypan blue dye esclusion method Rcwhs show that low eoncentrations of amwile AS are uny slightly tosic to human PAMs (n vitro. Hipher eoeceMrations, however, cause si6nifkant toskity. Phagocytosis begins immediately upon contact with AS: the PAM either wraps itself around the end of the fiber, ewdocyt'ain6 it by slowly moving toward its opposite end, or w spreads toward each end from the mdline, then completely engulfs it. The macropha6e membranes are also aAected, as re- /kcttd by an increase in cyroplasmie bkbbiag known as :eioais that probably resuhs from Uve ceM's high metabolic activity, and by the appearance of a (iMow-hke martrial in close approswnation with the AS fibers in the area of the PAM membr.ne whcre the initial contact is made. The precise nature of this cellular product is curremly under study to determine its biochemical structure Further research onto these biological interactions may prove valuable in establishing the etiology of AS-related lung disorders. McLemore, T., Coroow, M, Mace, M., Arnmr, AI. S., )enkins, T., Snodgrass, D., liLrrM, R., Wray, N., and Brinkley. B. R. Canrrr L.errers 6:1fi1•192, 1979. Other wFl.rt: Veterans Admioislration HospNd and the National Institutes of HcahA. From the Veterans Adminislratioe Hospital; the Department of tsiology, The University of Tetas System Cancer Center; M.D. Anderson Hoapital and Tumor Institute; and the Departments of Medicine and Cell Biology. Baylor College of Medici.e, Houston. IN VITRO ACTIVATION OF CIGARETTE SMOKE CONDENSATE MATERIAIS TO T/IEIR MUTAGENIC FORMS According to this report. both tAl and 2A1 reference eigarettes contain substrates for hepatic mrMxwsygenases capable of generating metabolic inter- medutes that are muu6eme /or S r2oAimrnam tester strains TAIS711 and 7 Aak the laure n select:vtty more unsibve to muu6enesis induced by smoke I condensate. The most muwagenic franions, nmrcover, are not the o.es that contain polrcrclic aromatic hydrocarbons (PAN) but Ihose supposeMy con- taining such base-soluble chemicals as aromatic amiees. The data supest, therefore, that most (about S/9i ) of the total emtagenie activity of theae co.- dcnsates lies in the briic fractions and not in the ones containing the PAH. In addition, reouse puimonary tissues seem capable of activating certsi. PAN and aromatic amines but not others. Mouse, rat and husnan pulrnonary liswrcs are currently being compared for Iheir ability to metabolically activate cigarette smoke condensate euKrial to biologically active forats. Kouri, R. E. tr .f. (lrfkroMolosir.f Atsocl.tet. Inc.) and /enelkl, IY. F. In: Water, M. D. r1 d. (eds.): AppIkwrions o/ SAorr-Ter.w Alo.r..ys M rAe Fr.rrlonaion o/ Compk. Endronmenrd M/trwti, New York: Pkawm Puk« Inhin6 Corp. 1979, pp. 497-512. From the Department of Biochemical Onoeolo6y, Microbiological Arociates, Ine., lletheda, Md.; and Childreri s Hospital of Los An6eks, Loa Angeles. ORGAN SPECIFICITY OF INDUCTION OF ACTIVATING AND INACT IVATING ENZYMES BY CIGARETTE SMOKE AND CIGARETTE SMOKE CONDENSATE • The effects of cigarette smoke and eigarette smoke condensate oo the activating and inactivating erxyme syalerws of the body were atudied ktn by in vivo and in vitro methods. In the rat, ciprene snake ieh.lation selaedisely induced lung and kidney aryl hydrocarbon hpdroaylase (ANH) activily. Oa the other hand, eposide hrdratase (EN) and Rlwathione S-traruferue wat trot si6niAcanty modi/led in any tissues o/ thtt keated animala. Coropwed to the kidney AIIII, the lung hydroaplase was three-fow times wtore sensitive to ammaR concentrations of cigarette stnoke and aeemel Io Aave a longer k~ioloRkal k,.lf• li/e. Is bolh tissun, the induced ANN presented the sanse /n virn se.ailiviiy to various inhikNors as a polycyclic hydrocarboo-itrdreed ANN. 1s the /n Wn+ uudies, cilareue ar.oke condensate fractions (CSCF) induced both ANN a.d EH activity in primary fetal rat liver ceM erltures. NeverthekesR tAe ANN activity responded faster and so lower concentrations of CSCF thas did die EII activity. A{ow conceMrMion of bewt(a)antlwat:ewe imWced only the AHII activity, while rrrnr-stilhene oitde enhanced selectively the EN activity. Appropriate concentrations of CSCF or of phenob.rbital determined a paraRel induction of bowh enrymes. 11 seems, therefore, that the liver cell cuNwe eonstitwes a unique tool for a comparatiw study of the A111/ ud Ell In- duction mechanisnss. The eaiVence of coordinated or independent biochemical control of AIIl/ and EN activity is discussed in this paper. Gielin. 1. t. et. d. ArcAiv.r of To.kolorT. Suppl. 2(Mechanism of To.ic Action on Sane Ta{et Organs). 2)9-251. 1979. Other arrporl: Fonds National de Is Recherche Scientifitiue Mldkda. From the l.aburatoire de ('himic Midicak, Instilul de Palholotie, Lille, Belgium. 1 16 17