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ANDREWS OFFICE PRODUCTS CAPITOL HEIGHTS. MD lKl

t IGINAL l 0~• I2EMOVl: UNLI:SS TO MAKE A l'HOTOCUPY _

S(:IENTIFIC ADVISORY BOARD to The Council for Tobacco Research-U.S.A., Inc. n of December )1, 1981 I.EON O. IACOBSON, M.D., CJfw~in+an lusrp/4 Rrtrnstrin Pro/cssor of Bir>loscul Sciences (emeritus) Professor of the DrportnKn( of Medkine (enrerinu) Univer:itr of Chicago Chicago. Illinois RICHARD 1. BINO, M D. hirn tr» of Erptr(en.ntd Cardiology and SriMti/ic DtvtloQTMt Huntington Medical Research lartilnte, Pasadena. California Pr.pfrsror of Mrdhlne (tnw.(Irs) (lniversity of Southern California School of Medicine Los Angeks, California ROSWEI.1. K. BnU'iWELL, PH D. Prrpfrssor of ()nrofrorr McArJk Laboratory for Cancer Researcb University of Wisconsin Madisoa, Wisconsin DRUMMOND FI BOWDEN, M.D. Professor and Head Department of Patholotnr Unrversity of Manitoba Health Sciences Center Winnipeg. Canada MICHAEL 1. BRENNAN. M.D. President and Medical Director Michirn Cancer Foundation DetroN, Michigan IOSEP(1 D. FELDMAN. M.D. MarnAn, Research Institute of Scripps Clinic Scripp~ Clinic and Research Foundation La Jo4a, California WILLIAM U. OAR DNER, Pw.D. E. K. Hunt Prolessor of Anotorny (emeritus) Yak I/nivenitr ScMW of Medicine New Ilaven. ( onnccticut ROBERT ). IIUEBNER, M.D. l.aboratory of Cellular and Mokcular Bio~ National Cancer Institute Bethesda, Maryland IIENRY 'P. LYNCI/, M.D. Professor and ClYafrwroen Department of Preventive Medicine and Public Neahb 0ei~hton Univeraity School of Medicine Omaha, Nebraska HANS MEIER, D.V.M., Dr. Med. Vet., M.RS.!!. Senior Sts SrlenNrt 'iThe Jackson Laboratory Bar /larbor, Maine GORDON 11. SATO, Pw.D. Pro%ssor of Biofoty University of C.lifornia, San Diego La lolla, California I SHELDON C. SOMMERS SdenN/ic INredo., The Council for Tobacco Researcb-U.S.A., l.e. Clinkal Professor of Pmlblotr Colkge of Phrsician:& SurSeoos of Cowatbh University New Yort, New York SeksUie Std.(Tt.e ('.weY SHELDON C. SOMMERS, M.D. Sdau!/k Dirercror ROBERT C. NOCKETT, PN.D. Retea+ch Director DONALD N. FORD. Pw.D. VINCENT F. LISANTI, D.M.D. Associate Research Director ~ Ansorla(t Research Director DAVID STONE, PM.D. Associate Rttearch Director J C.) /-+

I 1 CONTF.NTS Introductwa . . . . . . . . . . . . . . . . . . 7 Abstracts o/ Reports . . . . . . . . . . . . . . . . 9 Cancer-Related Studies . . . . . . . . . . . . . 9 The Respiratory System . . . . . . . . . . . . 25 Ileart and CirculNiow . . . . . . . . . . . . . 49 Neuropharmacolop aed Phrsiolop . . . . . . . . . 63 Pharmacology and BiocAemislry . . . . . . . . . . 71 Immueolo" and Ad•Qtive Mech.ais.u . . . . . . . de Epidemiology . . . . . . . . . . . . . . . . 95 Active Projects . : . . . . . . . . . . . . . . . . 100 Contpktrd Projects . . . . . . . . . . . . . . . . 109 Index of Priecipal in.eaigaton . . . . . . . . . . . . 122 Indet of Senior AulAon . . . . . . . . . . . . . . 123 i HANS MFIER 1929-1981 Ilans /Keicr, • a+ewrher of Ihe SciawtiAe Advisory tloard (or trs years. deed May 14. /lc Iw/ Ueew a Senior SIaA Sciewliq •1 The lack- sow LalaraNwy ia Uar Harbor. Maiet, rl eh he joined in 1%2, •aa a comuMael to the Nuiowrl Cancer IwsliMrle sincs 1970. H.w. waa a di.linguished cancer researcher wilh a spedd iaNeresl in viroloRy, a /kld in wh:rch he allaiaed aw Mlerwtiowd heppNMiun. Qu:e1 a" aoll- spoRen, he had greal warwNh and ernplhy •nd an aPqrecialive sew o/ hunwr. /le contributed r•isely and wweltishir le Ihe efforts of /lr ScieMiOc Advisory tloard and, iw hia owa tabm•1ory, worted " •d hard /or the resoMNion of one of m•aUwd's lntJW tiller diaeaae•. Ha will be onisaed as a persee and a scienlisl. a

I Introduction I The rcMearch progranr o( the Conncil for Tobacco Research-U.S.A. mo.ei ahead Mcsd,fy in 1901 wilti the emphasia codierriwg to be directed toward gaining additional tno.~kdge of esncer, cardiovascular diseases ad etiro.ie polmonary aAments. The Scienti/k Advisory Board to the Council closely reviewed scores of applicNions from irwkpenden/ investigators for research urppxt, and by year i end the aumher of original grants approved by the Coourcil had risen to 709. Councii funding tor the research prograrw since iu Incept:un ra.r staads at more than =69,000,000. (lrant recipient. published 144 documents ira 1981 that actnowledg[d Council suppHt; the taal of such p.htieationa is now 2.026. Abstraeta of the 1901 prhhcations are incluwkd is /his report. Several chantes occurred in the eumposition of the Advisory Board dda ing thc year. I/ans 1.. Meier, t).V.M.. died irt May. William U. Gardaer, Ph.D.. resigned as Scientific Director of the Cornrcil but remaHn a mentber of thc Board. lie was succeeded as Seien/i/le Director by Sheldoe C. Somwras. M.l)., a lons-time member who had Seen C'hairmsw o( the Advisory Board. Succeeding Dr. Sonwners as Chairman was l.ew O. )aeobson, M.D., a Board member since t9S4. Roberl ). Hrebser, h1.D., hecaeu a.nertrber ewwihr at the erwl of 1911. Two more d'winpii.Cled scientists joined the Board. They wers Drwt- mond H. Sowdew, M.D.. rro/essor and Head of the Department of rathobp at the University of Manitoba Health Sciences Center in Wiwripc& Ca.ada, and Michael J. Brennan. M.D.. !'resikat aed Medical Director of the Mich- iian Cancer Foundatioa in Detroil. 7

i Abstracts of Reports Following are abstraNs, approved by the awhors, of reports on sirw research acknowledging wpport from The Council that have appeared i. scie*- ti/k journah since publication of the 1980 RepoA. Tbs w.are of the tetiphat is in italics. 1 he ahstracts are grouped under these headinp: 1. Caswer•Related Studia, 11. lhe Rcspir.tory System. 111. Heart and Circulalion, IV. Nevropharmacololy and PArsiolotr. V. Pharnucology and siochemistry, VI. Immunology and Adaptive Mechanisms, VII. Epidemiolop. 1. C..neer-Refated Sewdiea I I I FAMILIAL CANCER IN AN ONCOLOOY CLINIC Ksowkalge of hereditary cancer symirorne identilkation, often haaed upon twnor pro/Mes, age of onsel and t)rrnor d'suibwiow within the kindrad, aid signilkamly in the diagnosw of eancer. As reported here, all eaaar pM tients (5651 trested at the CreiRAton University Aecokoar Clinia belwaes luwe 1978 and April 1980 were entered iMe a pospec/ive ra+wilT Yslery ascertainment prolocol. This consisted of an ieitial evaluation by a«{iMesd nune wi4ling both ~ueain.n.ire and perse.d (Merriew. T1he initisl aaseas- ment yielded 199 ()s.s9i) tamilin wilb 1wo wrore fami/r merrrbets with cancer (all sites) wilhin an iwfornwaliMe wtkar eorraponeAt, which ooolilMMd parents. Rrandparents, auMs/wc{a, sihlings, aed diWren. One or rraon of the operational criteria for cancer (anrilidily, "aw+elp vertical trarnmirim o/ cancer, bilalerality, and/or aardlipk prinwies, early age of onsel, and 1Mea or wwre site speci/{c canoers, were found on physician review i. 171 ( 70.3%) of the families. This group was referred (pr oomprehensive canoer Re.e1k evaluation consistinR of pedigree ealensiow and tumor veriAcuion tlraislr aR second degree aod, when possisle. third deRree relatives. 11 was dekrad.ed that appro:imarely 4% ol the 101l1 clinic population derwons/raled Mdinp compatible with hereditary cancer syndromes. The universal eatension of Ihese surveillance meihods in clinical practice is highly recommended here. Albuw, W. A., LywrA, N. T., Rec.baren, l. A.. Organ. C. H., Mailliard, l. A.. 81ack, L. G., Folk11, K. L., and Lynch, I. Cancer 17(9):2111•2115, 1981. OtAer w'/wrtr FrNCrn.l Order of Eagles. From 1he Institute for Familial Cancer Management and CorNrd, lnc., a.A Ihe [kparrmem of Surgery and /'revemiw Medkina, Crelghton Vniversllf School of Meditlne, tMuha, Neb. 9 I

I AMII IAI BRI-AST CANCER AND ITS RECO(:NITION IN AN ()N( O1 (K:Y ('1-1NIC This p,per focu.rs attention upnn in-depth clinicopalhologK studics of breast canaer pronc families wherein cancers of all anatomic sites were cri- trcally asesscd in contest with genealogy. For this study, lamily hislory of cancer was evahrated tor 79 breast cancer probands from among r series of cunseculrvely ascertained cancer paliewts undergoing trearment in the Creikh- Inn Sl l.seph's Oncotogy ('linie. C..cer prevalence lor e•ch family was quan- Idkd by using a statistic that accounts for variable size and age structure among families fo lesl the wuM hypothesis that cancer risk is independent of tamrly memhership, rhe dislribuliow of this statistic /or families (n their original configuration was eompared wilh /he drslnbulions obrerved when rclarives were randomly assigned to famifies in 99 random prrmrMariom of family nreml.ership lhe results indicated signifkant heterogeneity for cancer rnf< •nN.ng retariver of breast eancer probanJs, which suggests that the isola- turn of high rr.k families can provide • meaningful resource (or in depth studies in Measr cancer genenics. The uudy of breast eancer-prone familin harbors a pnwerlul potential (o. beNer comprehension of mechanisms of ear- crnotcnesn and tl.e development of novel pursuits toward improved cancer control. 1.rnrh, 1I T. er at ('anrrr 17 f t I I' 710 27 )9, 19!> 1. Other support: Fr•ternal O.der of Eagks. From the Crcighlon Univcniry School of Medicine, Omaha, Neb. ROUTINE CYlOD1AGNOSIS OF PULMONARY MAI.IGNANCIFS Material routinely sudbned frowr the tracheal lube of patients under general endo(racheal anesthesia for surgery is twrmaBy, ddiscarded. In this sludy, haweMer, the suctioned •ecretions trom 10.621 urch patients were pre- served, and later eaamined microscopically. Among these specimens. 11 q- tologic•lly abnormal smears were obtained /rom subjects with unsuspected pulmonary involvemenl, an incidetsee slightly more than 1:1.000. lbe aceu- r•cy of this method was assessed by calculation of the percent of abnormal smears obtained from patients wilh prediagnosed bronchogenic carcinomas: 40% when wctioned material was irnmediately spread on slides. and 67% when cellular concemration was achieved by mucolysis followed by filtration or cenlr//uta1NN1 In addilion, by using this lechniyue, a wide variety of cyto- narphnbSic and cylochemical changes in tracheobronchid cytology of both intrinsic and cstrinsic rnigin were discovered, chances which otherwise would have been ms%ard AhMwgh routine cytologic screening of all patients under- pant tc..cul rn.larahr.f .nnrhrs.a is not advocated here. it does seem L,w.,Mr itA.. rA.o nroA,.J t.-.W prn•r henefkr.l for the early deaten.wis of mal.to.nr n.,,/Ja.... .w a p,yrd.o..n ar rod I / to I ('hnfon, !. cr at. Arr hirei o/ ParholoKy s l.hor.oorr Mrdlclne 105:11-11, 1951. Other at.rport: U. S. Public Heahh Set.ice. From the (kpartment of Anesthesioloty, New York Unirenily Medical Ce.- ler, New York. ARE TUMOR MARKERS USEFUL IN D/AONOSINO LUNO CANCER? Becanse it has been no/ed that many hrng uueon have a Io+K latency period and a slow 6 row,h ra1e, the idea has been ad.anced that biochemical marken-ho.rnones •nd other metabolic producM of lumors--mi6h1 rewe•1 1h• presence of malignant disease .1 a very early stage and thus improve the .ur- vival ratio. The paper presented hero deserihes some o/ Ihese marker systems and evaluates them. In one ease, armed with a preeise and simple .•uy sy- tem, investigator. tested the hypothesis lhat elevations in peptide horwqne concentrations in blood might be a sensitive Iwdieator of the presence of a tumor. Tfiese and other studin have reveakd many interesting endocrinobok interactnwrs, but onlorlunately have not provided a su/Rciemly reliable and sensitive early marker of tumor presence. There are several other •ubstancp which have bera /ound 1o he presenl in abnormally high amounls in the blood or urine of patients with bronchial earcinown. Some ae producys of Ihe abs- plastic cells themselves and thus represent trut lumor marken. CareMoetw- brynnic actigen is the best sun/ied and uro.1 inleresting of tbese subslanees. Aryl hydrocarbon hydro.ylase and free seere/ory component of imrnunoklobu- lin A studies are ako touched apos here. From a diRerent facel of en/y de- Iection. fluorescent bronchoscopy, shows promise a• a means of very epdy anatomic identification of hmR tanan when the tunwrs are curable surgically but not deleclabie by traditional methods. MerriB, W. W. and RernWds, N. Y. Thr lorrnd of Resplrarory Diitarrs 2:1,-52, 1981. OtAer wr'prtr Ch.rk• E. Culpepper For.dallon- Prom the Department of Medicine and Internal Medicine, Yale U.Irenity School of Medicine, New Narew, Con.. RI:ASSSF..SSMENT OF THE RELATIONSI/IP BETWEEN ARYL IIYDRO('ARBON NYDROXYLASE AND LUNG CANCER In this study of the retati+nship between aryl hydrocarbon hydroaylw (Alill) and lung cancer, ANII levels were evhutcd in mito6ew-aclivNed lyrnphocytes cultured with and withoul 1ha inducer benzanthr.oena In 1hs medium, and in pulmonary alveolar macrophages (PAMs) that were /reaY) lavage.l from 114 cigarette smokers, 6) with and 51 without lung eanorr. Considered scpar•tely, neither lymphocyte Allll nor PAM AH/l kveia wen distinctly difterent in either noncancer or lung cancer patients. Ilowever, whew 11

I I canswl.rcd simulUnerMn/y, lymphocyte and macropha6e A1111 lcvcls werr yrule Jd/cren/ in rMmc.rncer and lung cancct pahents. '1he /ung cancer patient RrrNIr was seen to comain a si`ndicanlly higher percentage of petwrns wid, hrKh levels of A1111 than dw] an age-matched gnrrp of noncancer patients. l rnm another view, rcwlts of this strNly showed a 1:1 corrclahon of PAM and Iymphocyle Atilt kvels in noncancer patients and an absence o/ correlation of the trssuc A1/11 levels in lung cancer patients. Overall. it seems, on /he basis of the work presented here, that sint/e-tissue Atilt analyses are not ade- wsate (nr evahration of Atilt activiy in tang cancer populations McltnK+re. T. 1.., Manin, R. R., Wray, N. P., Caovell, E. T., and E'utbce, D. L. Cuncer Ie(6):11)11•144), 1911. (1tArr aurp.rrr National Lstilu/es of HeaMh, Amerieaa Cancer Society and the Veterans Administration Ilo.pital, l/ouston. /'rons the (kpattment of Medieine, eaybr College of Medicint, ifouslon; Veterans Admmistration Hospilal, Houslow; Department of Phrrmacology, lean College of Ostcop.thic Medicine. Fort Worth. and GenrlKs Center and IkpartmeM of Biological Scienoes. North Teaas Stale University, Iketos. A MFTIIOD FOR 1)E7F("11N(: ARYI. HYDROCARBON IIYI)RUXYI,A!l'1? AC11V1 t 1ES /N CRYOPRESERVtiD HUMAN 1.YMP/1()CYTFS A standard procedure for the frecring, thawing and culturing of eryo- ptesetved human lymphocytes ia pretcnted here. Using this rnetM>t/, three parameurs- -aryl hydrocarMrn hydroaylase (AIIH ) aclisNy, NADII-dependent cytochronse c rednctase (cyt c) activity, and 11111 thymidine (`ll-ldR) in- corpo.ation-were monirored in human lymphocytes erynpreserved for petiods up to .N+e year. Ihe kinetics for eapressiun of bersr/,.lanthracene-11/A1-in- duced Ali/l activity, cyt c activity and 'H-TJR incorporation were similar in freshly cultured and cryopreserved cells. for this suwfy, lymphocyte sampks from 10 individuals were cdlecteJ once a month during a Ihree-month period and cells were either cuNured at time of donation or eryopteserved for later assay. ResuNs Indicated that she cryopreaers,ed lymphocytes ef6cien0y re- sparled to mito6en activation. The inlra-;wJividual variation in AHH aetiv- itics was reduced in the cryopre.ers~ed Iymphocyles compared to the freshly cultured cells, and the relative canking of these individuals in terms of their Atilt activities remained constant fa both fresh and cryopreserved sampks. It seems, Iherefore, that eryopreservuion oAers si=ni8cnq advantages over the freshly cnitured lymphocytes because it aBows for lymphocyte samples to be collected in diverse geographical locations and over eatetMkd perinds of time and yet permits for the culture and assay of all the cell samples at eaactly the aame time. Koati, R. E. cr.f (MkroDiofolkaf Arsoc(arcr) Cunrcr lstarr 14 29 10, 1981. F'rr.m the [h.rsr.n .d lo..coi.ty and ()ncolup. Microhiological Associalea, Ikrhe.da, h/d 12 SI(:NII~I('ANf VARIAiION IN MOUSF.-SKIN ARYI. HYnR(X'ARBON IiY1)KUXYLASI: 1NUUCIBIL) I Y AS A FUNCIIUN UF fNE HAIR (iRUW I H CYCI.E Skin, which is a goud biological model tor the study of polycyclic hydro- carbon mclabolrsm, eshibits three major morphological phases in she hair growth cyck. In earlier works, it had already been demonstrated that akis possesses activating and detosilying enrymes, notably aryl hydrocarbon by- Jrosylue (Alilt) and eputide hydro/ne. For the ptesent .tudy, aw atnitpt was made to determine whether Atilt activity and i1s irnhtcibilily vatied dur- ing the hair trowth cycle and could possibly eaplain cectais varialiona in tlin sensNivily of carcinogens. An easy, aapid and improved /eclwsipue (or hoew IenirinR whole skin was devixd for this work. Using the new homoRcmirj.R method, it was shown that skiw AHH activity in C37B1.J61 and C)Hlk,o mice could be induced by i p. injected or topkaRy applied methykho/anthrtwe during a defined pesiod of the hair growth eyck, /.e., between the Mth a.d 1hh Jays after depilation (Stage 6 d/he anaprt phase). 1o each eaperi- mental makl, there was an optimal methykholanthrene eonoeMralioa which yielded a maaimum induction. 'Topical methykholantArene was also respow- .ibk for a smaller Atilt induction when the chemical was applied the aamtr day that the club hain were pkucked. On the oller fand, skin Atilt activity was never induced by methykholamhrene in OSA/2t mice, a genetically ao.- responsive strain. from a Wsieological poiM of view, the fact that akia ANN activity could only be Induced by polycyclic hydrocarbons at a certain time during the hair growth cycle of a fieneticaNy responsive attain might be o( great imporlance. Manil, l-., Van Canlfort, l., l.apit re, C. M., nad Olrlen, l. E. ertrish lournd e/ Cwctr t7:210-221, /991. From she lsbo.atoire de Derma/obsie atd Iabora(oMe de Chimk M1licdt, Institut de Patholostic. Univecsitd de Lifte, IJiile. Selgiww. CYTC)c'IIROME P-4S0 MONOOXYGENASE ACTIVITIES IN HUMAN AND RAT LIVER MICROSOMES Only limited data .re available a(he poeaewt time on the biochemical properties of the human enzymes. Nowe.er, recent studies have ahow. t!M suitable liver samples ooutd be obtained /rom kidney transplant dowa>< srd the microsomes used iw this study were ptepaled from human liven .etiuiri.d from renal donors of various ya and both seaes. Thcir druR-melabolkby capacity was measured and compared to that oI rat liver wuccoao..es. Tha following parameters were imestipted: eylodlrane P-ISO, crlochrome iS. NAi1P11-cytoclwome c reductase. eposide hydrolase, eryl hydrocarbon hy- drosylase, betwphetamine W-demethylase, p-aiitroawisok-O-demelhy/aae, ethoay/- cownatin-O-deethylase, slecoid-l6.-hydroaylase. In addition, the melabolirn of benro(w)pyrene, progeslerone, prerneeotone. te+toateront:. dehydroepiaw- drosterone. and estradiol was studied in deuil M.lrr.. RewRa showed dut Ihe cyluchrome P-1S0 content in the tivea of the kiJner transplant do.wrs was 10 2 t 7.9 nrnolt6 o( tiaure. Is did no1 di//er with /he aea of the de.oc. Comparison of the data obtained wah humaa and ta microaornes denusnacated qualitative and quantitative diAererrces which wtied with the pararneler. 1)

studied Ovtrall, results of these sludrcs indicate Ihat the drug-melaholizing rnryme activities (ethosycoumarine deethylase, benzphetamine demethylase, aryl hydrocarbon hydrosylase) vary as a function of the cytochrome P-IS0 content. The steroid hydrosylase activities do not fluctuate in a similar man- net. Speciflcally, this study shows Ihal the use of livers from kidney trans- plant donors is a promising tool for Iesting the metabolic pathway of new drugs in man or for scrcenin6 potential /o.ic or carcinogenic properties of iscw chemicals. Kremen. P., Beaune, P.. Creski, T.. De Oneve, /., Columelli, S., I erout, 1. P., and lae/rn, l. E. Errortan lo.rrwd o/ florAriwlm7 1111:399-f06, 1911. Otil.r .rtpHr Foads de Is Recherche Scleslifique MEdicak (Belgium). Fronm the I aborato'ue de C'*iwsM Mbdkak, Imlilut da Pathologie, l)nivenilf de Little, I.kge, Belgium, and FaeuNl de Mtdieine, IlopNal Neeker-tnfauts Malades, Paris, France. ONTOGFNIC DEVEI.OPMENT OF STEROID 16 a-HYDROXYLASE AS A TOO1. FOR THE STUDY OF THE MULTIPLICITY OF CYTOCHROME P430 This Mudy, ffollowed the evolution of sleroid 16 .-hydroaylase in the rat liver from birth to adulthood. To do shis, the quantitative and qualitative propanin of the steroid hormone-wxtaboluins system during the period of ontolgenic de.ebpment of Ihe animal wen investigated and compared 1o those of the adu/t nsoooosf6crraae sys/em. In the beginning. activities of progesterpne, teslosterone, preg newolo.e, aed dchydroepiandrosterone 16 .-hy- drosrla,e were undetectable in the fetal rat 1'wer. During the neonstal period, howe.er, the four erwymic acti.itiee increased in parallel so the carcentralion of ertochrome P-.30. Until puberty. 1" developed similarly in male and femak rat livers. From the I0th le 1he 3S1h day. Ihe four steroid 16 .-hy- droxyfase activities increased rspidly M the male rat liver, but no/ in the female. T)te sesual differenlialion of tht steroid 16 rhydrosylalioe observed here look place around the 55th day. 1Mheo the sduh rat liver was studied, steroid 16 .-hydroaylase was supporld by two fonws of eytochrome P-4S0 (forww I a.d 111 which differed i. thck relative affinities for Mte various steroid subdrales and by Meir relative proportbas iw mak and female rat liven. The transition from the bnm.lure so the adult rcpartilion of the 1wo forms ocewred during puberty md was correlated with the seeual differen- Iiatiow of the steroid /6 0-hydroa}Ine activitia. In another phase of this study, the /n dtro interactions between benw(.)pyrene and slcroids were oornPared during the eritical phases of the rat ontogenic devdopment. Pnkau, F.. Kolodtici, C., Krcmer, P.. Bad Cl.ten. !. E. Eurorron lo«rnd o/ SiorAtwrbrr7 120:213-220. 1981. OtAer support: Fonds de la Recherche Scientifique Mfdicak (Belgium). 1'rom the lAhoraioire de Chemk Mldicak, Institut de Paihob&. Univenitf de l.ilgt, 1.it1e. Belgium COMPETITION BfTWF.FN BF.N2O(a)PYRENE AND VARIOUS STEROIDS FOR CYlOC11ROML' t430-DEPENDENT RAT 1-IVER MON(X)X YOENACFS Many previous reports have described the eaistence of various types of in ritro competition between nwrao.yRenase aubarNea. For the present paper. the interaction lw vitro of steroids and bento(.)pyrewe (BP) was studied a1 the level of two rat liver monooRypenmes, steroid 16 rhydroaylase and ary1 hydrocarbow hydrosylsse (AHH). The resuNs obtained in these tests wuesl the following conclusions: (f) Steroid-/6 .-hydroaylase is partially supported by a speci/k cytochrome r-130 form which is s+o1 inhibited /n r'Hro by eto- Renota substrates. Steroid-16 .-hpdrosylase is oompklely independent from cylnchrome P,-130 (or r-IN11, as i1 h insewsilivq /n .Ino, 1o .-staphtlrs flavone; (2) AIIII is supported by two cy/ocfuorwe t'-130 forms: a speciRc form which is inducible by wtcth7khdaMMeae t>wd inAibited /n rhro by rnapMhaRavone, but is insemilivt to metyrapons .ed alaoids: and anotber kss specific form which h inhibited by nsetyrapoms and steroids /w .irro. It seean reasonable on these grounds so auwne the eaisterice of speciAc eyls- chrome r-4S0 forms that are responsible for endoRenow compound meubol- ism, or at least for hormonal steroid 16 rhydroaylation. This observation may possiMl he of physiological importance, as it could preserve endogenous metabolism Irom competition due lo enviromnenlal senobiotics. Paskau, F.. Kremen, P. and Gklen. /. E. CArnNro-diolotir.f lntenartioru 11:279-2116, 1961. OtAer a.rrprts Fonds National de Is Recherche SeieMiAaue Mtdicak. From the l.aboratoire de Chimie MEdicak, Inslitw de Pathologie. Udvenk6 de L%lge, Litge, Belgium. METAflOLIC INA(.`TIVATION OF MUTAQENIC DENZO(a)PYRENE METABOLITES: SIGNIFICANCE TO CARCINOOENICITY AND IMPLICATIONS FOR !N VITRO TESTs In lbis very weR-ressoed P.per. the twt.Renkilf. ..d /n sdtro IestinR of bsoto(.)prrewa (RP) tre ooasidered fretwaol dilleftest aspects. It is knowo that BP lus a Complea wrclabolistN involving rwa.r st zrmcs and, because of dris. complete metaboNsi.l sfstews for tbe aelivatian of BP ud its prennNalgenic metabolMea were wed iw the bacterial tnW.- {enkity leas presented here. The activatity .pslem. Ined wera freshly iso- lated intact bepatocyles or \onwgets.kes of their eelis wplleme~ted wit\ wr- ioueco(ador systems. '[he compounds Iesled. wete Ihe csei.oResr BP :d ( t)tr.nr-7,edihydro.y-7,>f-dihydro-Or (7,1-diol) and the rery weak car- cinoftens or tumor initiators l-hydroay-SP ( I-OH-BP), 9-hydroay-11P (9[H!- BP) aad ( i )tr.ns-9,10-"ydroar-9.10-dihpdro-§P (9,10-0iol). (AN .rs strong wwta6ene in the Arses lesl.) Results of 1hese lesu showed 111M RP was no/ mutasenic when tested directly or in the pteaence of hepaoeyts fwn.oe - enate. Ilowever, it became strongly muwatenic when an NADPH-genetatiae system was added to the homogcnale or when whole hepatocytes were used. T4 dose-response relalionship difiered matkedly under the two situations. 3-011-BP, 9-OII-BP and 9-10-di4g were not mWa6enic in the preaeoee of IS 14

r homofiena/e. However, when an NADPH-genersling system was added with Ihe cell homogenate, mulagenk eAecls were observed with all three. On the o/her hand, 7,tidiol was aetivaled by both homogenized or Intact hepatocytes to a very potent mwagen. Overall. the rewl/s of this study show that it is possible to activate BP and BP-metabolites to bacterial mwageos with intact hepatocyles. The mutagenkity often is weaker and the relative potencies of various compounds reatly diQerent from resuhs obtained with NADPH-for- tiAed cell homogenate. Afso, it was shown here that addition of eolacton of further enrymn coesiderably, aaltered wtwa:ewkhy. The alterations differed with different test compounds and the results became rrare similar to resuhs obtained with intact cells. Whee carciwornicilr was taken into accoual, it was seen that use of intaet hepaloqus instead of NADP11-fortilkd homoll- enale considerably improved the correlation of n/ulatenacNr with whole Mi- Ina) earcinopenkAy. Now, while intaet hepMoertes ma7 well awt be the opdmal metabolizing tqntem In sbort 1ern. tests for earcinofenkity and mutsgenicity. Ihis study shows that Iha Inetabolking system can very strongly affect the result of a ler. Therefore, if Inaclivation oocron in al.o it is reasoaabls to take the inactivating systems (Mo aeeounl also In /n vitro ters. Olan, ll. R.. P1at1, K. L., Voilel, M., Itader, M., Billinp, R., and OezcA. F. In: Itolmsledt, B.. Lawrerps, R., Mercier, M., awd Roberfroid, bf. (eds.): 1tIefAlMGmt o/ ToaleUy, aw/ N.ta.d Erdrtetlon, Elsevier: North Hollaod Biomedical Press, 1980. pp. 111-186. From the Pharmakolofisches 1..1itw der UsivenitH Maitut, Federal Republic of Germany. RAT LIVER CYTOPLASMIC DIHYDRODIOL DP-/IYDROOENASE: PURIFICATION TO APPARENT 1oOMOQENEtTY AND PROPERTIES tn this biochemical paper, a method is described for purifying a diby- drodiol deh7dro6enase 1o apparent harw6eneit7 from the cytoplasmic frae- tion of rat liver 1wmWnak. Some properties of the pmiAed peparation, and the eRect of this entrme upon tbe wwta6esicit7 of beetto(.)prrene arn also oonsidered herr. SpeciAcaMy, the pnrilkatior involved (NH,),SU, fractiona- tim DEAP.oelluk+ae ehromuography, hwerfaeW sahitK-M and gel Altration through Sephadea 0-100 tuperAne. The end product, which .a puriAed over S00-foid with a yield of about 14% whe* compared to rat liver 10f",000 It A wpem.lant. was Judged to be horno=e*eolas by several erileria. Phys.cal stud- ies suggested that the protein was a monomer with a mokcvlar weight of 13,000 and one NADPH binding site per molecule. Amino acid analysis showed IhN the enzyme had a relatively bigh content of acidic and neutral amino acids irn agreement with its isoelectrie point. Apparent Km vahres for benrsne dihrdrodiol and NADP+ were found to be 2.2 mM and 7.7 rM, nrspectivel7. Substrate specificity studies showed Ihat, in addition to benzaw dihpdtodiol, the deh7dro=enase oo.ld oxidize acewapthenol ad the 7.-hrdros7 group of steroids. No activity was observabk with a large number of other hydrosylated steroids possessing hydroay groups at positions lp. 11p. 17., 17Q, 20.. 20/1. 21 and 22 of the steroid skeleton. Furthermore, only sleroids which contsined a l keto group and no double bond at the A• position were reduced. 7Uis, and the fact that a range of nonMeroidal vkin.l dicds did not 16 serve as substrates, indkates a relatively narrow substrate specificity. The role of the enzyme in the metabolisnm of carcinogenic polycyclic hydrocarbos Is discussed. Vogel, K., Bentley. P., P1Nt, K-L., and OesrA. F. 77)e Iorrnd of eiolopk.f CArmi.try 255(20) :%21-9625, 1950. Frorn the Pharnwkologisches Institw der Uttiversili/ Maint, Federal Republic o( Oertna.T. QNZYMIC IfYDROXYLATION OF BENZO(a)PYRENE AT THE 6-POSITION BY VITAMIN K-HYDROPEROXIDI3 AND RAT LUNG IIY DROPN ROX IDASf} of /n henzoth(is.biochrene )pyrmie(BP)d p.atper,thethe6-posrokitionb o of vitamiwr.elaed K, is rhewilhMe hydroto.iativrldatiw e formation of vitamin K,-hydropetotide and its subsequent reaction with BP in the presence of soluble rat lung hpdroperosidase. Furthermore the chee.- kal synthesis of vitamin K,-hydroperoaide and the dual' inhibitory adioa o( thiodione (menadione-glutathione adducl) both as aw aryl hydrocarbow by. droaylase inhibitor and hydroperotidase inhrTritor is presented here. The apec- trd characteristics of vitamin KI-hydroperoaide and the 3.6-BP dione tft al.o shown. It is generdly t.own tha the identiAeatio/s of a proximate ear. eiro6en th.t results from the metabolism of eueilwAenic polycYclic hrdtr- earbom is a necessary prerequisite i.w order to stttdy the mechanism of chcw kd carcinogenesis. Studies are eowsidered in tbis paper that deal with the roie of dihydrodiol epotides of OP, eywchrorne P-4S0-448-Independeru reae. tioas, and hydroperoxidase reaetiows i. the 1lpdroaylaliow of <IP a the 6-posi- tion in chemical e.rcinoitawesh. Speeifkally, ahe studies reported fultp herm demonstrate that the mnem.Nr hrraA hpdroperoxidase is capable of «adift with potenliaNr naturally occurring brdroperoaides, e.p., vitamin K,-hrdro. pero=ide, to form the earcinoAe.6-h7dro>iy BP. Sloane, N. Iewbiorlr.ll(4):267-274, 19/1. From the DeparlmeM of Biochemislry. University of Tennessee Collqe of Merlicine, Memphis. DIALKYLNITROSAMINE s1OACTIVAT10N AND CARCINOOENESIS 71as nwni-review presents a sy.opsis of studies on the rnech.ninws of metabolic ac/ivation and carcinornesis of diaKytnitrosaR+ises. Since tha aiw- pkst and most cornmo. dialtylnilrotamine is dimethylnitros.mine (DMN), it was studied Arst. The considerable number of studies that were lakr ear- ried out on DMN are reviewed in the Arst section of this paper; sections two and three are devoted to the actions and reactions of dietbylnilro..mi.es (DEN) and methykthylnilrotsmine (MEN) and higher nitroaarninn. As overaN eonsideration of these many studies led to the conclwione that: (/) Prior metabolic activation of nitrosamines by various oaidases to alkylallng 17