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SCIENTIFIC ADVISORY BOARD to The Council for Tobacco Research-U.S.A., Inc. as of December 31, 197R SHELDON C. SOMMERS, M.D., Chalrtnms Direcw of [.aboratorles. Ltoos HiU Hospital Clinkal Professor of Pathobp Colk~e of Physkiana fc Sutgeorr o( Columbia University New York, New York RICHARD ). BINO, M.D. Director of Cardiology and lnt. snard MrdkJne Huntington Memorial Hoapitd, Pasadena, California Pro/essor of Medicine University of Southern California School of Medicine Los Angele., California JOSEPH D. FELDMAN. M.D. Read, Department of Immunopathology Scripp~ Ctink and Research Foundatbn La )olla, California WILLIAM U. OARDNf'R. Pw D. Sclenti/ic Director. The Council for Tobacco Research-tI.S.A , Inc. E. K. Nunt Pro/essor of Anau„ny (rnuritur) Yak University Schocd of Medicioe New Haven, Connecticut ROBERT J. HUEBNER, M.D. Chief. Laboratory of RNA Tumor Viruses National Cancer Institute Betheada, Maryland LEON O. IACOBSON, M.D. Joseph Regenste(n Pro(rstor of Biolotkal Scirnces Unive of Chicaso Chkago, Wnoia HENRY T. LYNCH. M.D. Professor and Chairman I Department of Preventive Medicine and Public Health 0eighton llniveraty School of Medicine Onaha, Nrt'rarka t HANS MEIER, D.V.M., Dr. Med. Vet., M.R.S.II. Senior Staff Scientist The Jackson Laboratory Bar Harbor, Maine LEE W. WATTENBERG, M.D. Professor of Pathology Department of Laboratory Medicine and Pathology Umversity of Minnesota Medical School Minneapoli., Minnesota JOHN P. WYATT, M.D. Director Tobacco and Health Research lnatilute University of Kentucky Lexington. Kentucky Sdenliie Staff of 7Ue Ceuneil WILLIAM U. GARDNER, Pw.D. SdenH/ic Director ROBERT C. HOCKE7T, PH.D. Resewrh Director DONALD tl. FORD. P41.D. VINCENT F. LISANT7, D.M.D. Associate Research Director Astoclate Research Director DAVID STONE. PH.D. Associate Research Director ,

CONTENTS Introduction . . . . . . . . . . . . . . . . Abstracts o[ Reports . Cancer-Related Studies . . . . . . . . . . . The Respiratory System . . . . . . . . . . 1leart and Circulation . . . . . . . . . . . Neurophannacolo6y and Physioloq . . . . . . . Pharmaeolosy and Biochemistry . . . . . . . . Immunology and Adaptive Mechanisma . . . . . Epidemiology . . . . . . . . . . . . . . Active Projects . Completed Projects . . . . . . . . . . . . . . lode: of Principal Investisaton . . . . . . . . . . lodes of Seoior Authors . . . . . . . . . . . . . . 5 . 7 . . 7 . . 26 • • 41 . . 49 . . 62 . . 70 . . 77 . . 83 . . 92 . . 103 . . 104 I ' Introduction This report m.riu the completion of 25 years is which 7b Council for Tobacco Research has supported a program that has bewrne sb" world's mosl extensive noa-governmeatal research effort rekvaet Io smokins and health. When The Council was estaMished in 1931, it adopted a basic policy that has been followed without deviation: to pl.ce responsibility for research policy and programming in the hands of a Scientific Advisory Bo.rd and to have 1ho research conducted by independent investiplon in tht3r own lnstitutions with so striop attached. Up to the end of 1978, the program has resulled in the publication of 1.622 reports and articles that acknowledged Council support. 77rua, the pro- gram has produced considerable data related /o smoking and health during a periud Ibat also saw Ihe generation of oomiderable eoMroveny and unotior about the subpct. No one can fully predict at this time what these publications will meau in the search for the answers b the problem of the major, a8ing-.»ocialed and constitutional diseases of eancer, heart disease and chronic pu4aoa.ry ailments. However, the hope is that some day the Andinp from one or mrore studies may provide keys or links for a major biomedieal advance. The Council eaisu today because the complex etiology of these oonslitu- tional diseases remains unraveled. These diseases have been associated ststiL- tically with amoking, but such a»ociations are not proof of cause and effect. The diseases are extremely complex-cancer itself, for eaampk, b siol os disease but many-and /indin8lheir causes will not be easy. The overall importance of genetic faelors is being increasingly reco8nired, accepted and studied. Progress is being rwade, but it is necessarily slow sad painstaking. The statement of many years ago that we live in a "sea of careLa{ear' is supported by nmch recent and current research. Many Ihin8l--both natural and man-made-that we ea1, drink and breathe have been implicated (n canoer and heart disease, at least in laboratory eaperimenls. But exact proof, proof that meets scientific criteria, is often lacking. Whatever contribution lo scientific knowledge that may be attributed to The Council's program, as evidenced in part by the numerous reports in the Gterature, is due principally to the members of the Scientific Advisory Board, p.st and present. 7lrese scienliMs, who have always maintained their inuilo- Iiond alfiliations, have given unsdOshty of their time and talents. 'i'he result of their combined cRort lor so many years is an Imaginalive undertaking that has enabled many reaearchen to pursue ideas and theories that might other- wise have gone une.p{ored for lack of support. lbe Council is gratified at the response through the years of Ihe sckn- tists who have applied to it for research support. 'Ibis intereat remained high in 1978 aand the number of requesu for support of new and continuing re- search assures a worthy future. 7 his report contains abstracts of research findings published in 197• that aCkMtwtrdeted ('ouncd sponsorship. As can be aeen. Ihe major em~hasis c.xr- linucs to be directed loward cancer, the cardiova+colar system and Ihe respira- 3

tory tract. The abstracts of the published papers are indicative of the areas of research supported by llre Council. lhe Council looka back with utisfaclion at what has been accomplished in the lasl quarter-century. It looks to the future with optimism and hope for research that will help sdve the mysleria of canter, heart disease and chronic pulmonary ailments. The Council pkdgea to continue its support of inde- pendent investigators in their effotts to add to scieotific knowledge related to smoking and health. I 1. ('.awcer-Relwted Studies Abstracts of Reports Following re ab.trac/s, approved by the aulhors, of reporu oo .ew research acknowkdging wpport front TLe Council that have appeared in acka- tifk journals since publication of the 1977 Report. The name of Ihe recipieM is in iulica. The abstracts are grouped under these headings: 1. Cancer-Related Studia, (1. lhe Respiratory System, 111. Neart and Circulation, IV. Neuropharmacubpr and Physioloty, V. Pharmacology and Biochemistry, VI. Immunobp aad Adaptive Mechanwns, VII. Epidemiology. CRITERIA FOR Se1.ECi1NO CHEMICAL COMPOUNDS FOR CARCINOGENICITY TFSfINO: AN ESSAY The ever increasing number of organic compounds conslantly being bstra duced into the various aegmenls of the wrorldwide economy cnrales 1he aeed b devise a formal scheme of selection criteria for testing of suspectod car- cinogens, and to establish priorities /or auch testinll. Four caleitoriea of erileria must be considered: (1) structural. (2) operation.l, (3) "guilt by arocia- tiwr," and (4) "after the fact." This essay Rives an analytic discussion of each of these criteria. Structurd criteria are deduced from a=eneral perspective of the structure-activity relalionship of known earcitsoRens. The operalional erih teria, which are complementary lo the structural ones, represent the sum of the bio-/unctional capabilities of chemical compounds. These capabilities oor- relate with the ability to induce malignancy lhroukh such mcchanisma as, for eaampk, mulajenicily, induction of DNA repair, and Mnmuno.uppression. The "guilt by association" criterion alates that a compound belonging to a chem- ical clase already known to contain several other compounds eslablished as potent and mullilarilet carcinogens should be 1esled under more stringent con- ditions than those of the standard bioasay. The "atter the fact" criterion deAnes previous epidemiologie indications as the basis lor selecting certain chemical agents /or carcinogenicity tesling. The overall significance of Ihe bioassay dala obtained on individu.l eompounds, however, should be ea+- sidered in conjunction wilh other endogenous ud eaoaenous interacting lac- tors such as: /1) synergistic and anlakonistic sKtivoy between multiple car- einokenie agents; (2) dlects of nonc.rcinotenk environmental agents on carcinogenic activity; and (3) viru.es, nusrition, radiation, age, stress, endo- erine bdanee and th, state of th. (rnnwne wrveillance system. 'fln huae number of akenu and facton, their interactions and the coesidence of fre- quently conflicting societal soala, indicate the seed for the establishment of versatile acientificoletal standards and guidelines applicable to eatetoria sarl types of chemical agents rather Ihan to individual eompounda. ~ /rror, l. C. ~ lorrnd of Ewrironm.nrd laholosy ad Toakoloty 1:1))-ISe, 1978. 7' 6

OtRer aupportr National Cancer Imtilute. From the Seamen's Memorial Research Iaboratory, U. S. Public )ler,lth Service Ilospilal, New Orkana; and the Department of Mcdicioe. Tular c Medical Center. New Orleans. UI:1 RASTRU(`Tl1RAL AND METABOLIC DETERMINANTS OP RINISI'ANCE TO AZO-DYE AND SUSCEPTIBILITY TO NI1 ROSAMINE CARCIN(7GENPSIS OF THE GUINEA-PIO The a:o dye rcductase and nilroaamine dealkylaae activities of normal guinea-pigs and rala were compared b those of animals fed ar.o dye and niuo.amine. Electron microscopy reveakd ultraalruetur.l alkralions of the guinea pig liver during nilrosamine administration. Dielhylnitrosamine (1)EN)- induced hepatic lumot cells showed extensive proliferalioo of the rough endo- plasmic reticulum ( ER ), while the smooth ER wat quite tp.rx; the opposite was true in the prema)ignanl liver. In the rnt, however, administration of either DEN or Y-methy/-Idimelhylaminoa:obtnzene (Y-Me-DAB) caused prolderatioe of the smooth ER and sparsity of the tough ER in both pre- malrgnanl and malignant tissue. In both specin, the number of ribosomes on the outer surface of Ihe liver rough PR was greatly reduced as was the RNA/ protein ralio, which correlated with a decreased r+eaponse to an Slt probe In nmicroaomal suspensions A:o dye reduclase activity was higher in untreated rats than in untreated guinea pigs. After sit weeks of T-Me-DAB feeding, however, this was 76% lower in the ral while there was no significant decrease in the guinea pig, which is refractory to azo dye carcinotenesis. 77sus, as in- geslinn approached the lumorigenic threshold doae. the rat liver's ability to inactivale the dye was much impaired, but not that of the guinea pig. Control levels of ni/rosamine dealkylase were idenlical in both apeeies, however, and remained essentially unchanged Iter 10 weeks of DEN adminislration. Since nitro.amine dealkylation represents activating metabolism. this provides the basis for the idenlical susceptrbililies of the rat and the guinea pig to DEN carcinotte.rcsis. Of the two enzymes eaamined, only Ihe guinea pig azo dye reduclase appean to be independent of glucose repreaion, since its activity was unchanged by starvation. Statvalion-irduoed elevation of azo dye reduclne activity in the rat was not affected by Y-Me-DAB administration and only slightly influenced by DEN In both specks, however. DEN abolished the aarvation-induced increase of nilro.amine dealkylation, while )'-Me-DAB decreased it only slightly. Bryant. (i. M., Sohal, R. S., Arpn, M. P., and Arcos. l. C. drittrA Joucwd of Cancer 76:676-691, 1977. Other .u pport ) National Cancer Institute. Prom the Department of Mcdicine, Tulane Medical Center and Seamen: Mernorr.J Kcseaah I aN.ua1ry, 11 S Public Ilcalth Service llolpilal, New Ur1c..ns,..uJ Ihc Ikp.rrotcnt of Itwhity, SanUhcrn Melhodnl llurverslty, 1).IL1. I IiYDROCARBOMNrI'ROSAMINE PULMONARY SYNCARCINO- GEN['SIS: RI:CIPROCAL EFFECTS ON METABOLISM This study reports on the entynqlogical correlates of the pulmonary ayn- carcinogenesis which h.a been observed between melhykholanthreoe (MC) and dimethylnitrosamine (DMN) following acute administration of the car- cinotenr. E.perimenlally, .ryl hydrocarbon hydroaylase (AHH), eposide hydrase. DMNdemelhylase, and ilIvlalhione-S<potide Irana/eraae activities were studied in the liver and lung of DBA/21 and C378V61 mice. two atraiaa which are, respeclively, noninducibk and inducible for hepatic A1111. Change in theae enzyme activities was determined following acute adminislralioa of MC. DMN, and their combinations. In the liver-eacepl tor a.ubstantial ie- duction of hepatic Atilt in the C37B1./61 alrain-1he metabolic pattern is very similar in the two strains. In both slrairr DMNdemelhylase ia highly sen.utive to DMN prelrcatment at the aia mg/tti level. Increase of DMN to the total syncarcinogenic dose of 60 ms/kil abolishes induced AIIH in the C37B1./61 and brings about substantial inhibition of aB other en:ymea studied. In the lung of the two alrains, induced Atilt was found to be highly aensitive to DMN pretreatment in the DBA/2J, and DMN-demelhylaae 11 (the only DMN-demethylse present in the lung) was substantially reduced following Prelrealment with 60 mg but not with 6 mg DMN. Tlse most interesting re- sulls in the lung are .een with the C37BL/61 strain since, with 60 mg DMN/ tg, there is a substantial decrease of the epoaide hydrase together with a substantial increase of the AIIN. This suggests a mechaniatic interpretatiom of the Atilt increase within the framework of the known palhways of hydro- carbon metabolism and a greater suaceplibilNy of the C378t./61 strain to Ur combined administration of MC and DMN. Arcoi, J. C. d at. In: Gelboin, II. V. and T: o, P. O. P. (eds. ): rotpcycflc Hrdrocarbowr md Cancer--Environnrent, Chemistry and Alttaboll.ns, New York: Academic Press, 1979. vol. 1, pp. 271-282. From Tulane University School of Medicine, New Orkana. DIPFP.RENTIAI. EFFECTS OF a-NAPTHOFLAVONE AND PREGNENOLONE-16a1:ARBONITRILE ON DIMETIIYI.NITItO- SAMINN-INI)ll('I:D IIEPATOCARCINOGENFSIS 7he carcinogenicity of dimethylnilroaamine (DMN) is inhibited by the co-administratiun of either 7-melbykholanthrene or anminoacelonitrik, com- pounds which substantially lower the activity of DMN-demcthyla/e, a micro- somal mi.ed-lunction oaidase that catalyzes the activation of DMN. In the present investigation imed at elucidating further this correlalion, the eRect of administering jil-naphthoflavone (0-NF) and pregnenolone-16.-carbonitrile (PCN) on the hepatocarcinotenicity of DMN in male SD rata wa%* explored. Bo1h /! NI: and 1'('N are potent represson of the bw-Michaeliu-conslant en- :ymalic form of 1)MN demethylase. Ncpalie tumors were found in every group of rats receiving DMN alone or in combination with A NP or PCN. 8 9

I I I a The morphology of these tumors waa quite similar for all three kroups, with 3396 of the 53 liver tumors observed being angiosarcomu. DMN-induced bepalocarcirw6enenis was partially inhibited by PCN and enhanced by P-NP. Seven liver turnors were found in 45 rats fed DMN plus PCN compared to 32 liver tumon in 43 rats fed DMN plus aNF; 14 liver lumon wrre found in 43 rats fed DMN abne. No liver tumon were detected in rats that received only PCN, /f-NF, or Ifse administration reAicks. The unespected enhancement of DMN Induced carcinokenesis by aNP prompts a reexaminatwrn of the presently accepted activation mechanism of DMN. Argu., M. F., Hoch-l.igeli, C., Ars•or, !. C., and Conney, A. H. /orrnal o/ the Narional Cancer Iwsrhr/e 61(2) :441-449, 1978. O1Mr arpportlt National Cancer Institute. From Ihe Seamen's Memorial Research Iaboratory, 11. S. Public Hcalth Service llospital, and the Department of Mediclne, Tulane University School of Medicine. New Orkans. USE OF HInH CONCENTRATIONS OP DIMETHYLNI71tOSAMINE IN SACI-I!RIAI. LEl HAI-1 fY, MUTAOENpSIS. AND flNZYMOL(X)ICAL STUI)I PS Previous studies have established that the potency of dimethylnilro.amine and dielhylnitrosamine in bringing about conformational changes in protein is comparable to that of urea and kuanidinium chbride, well-known protein- denaturing agents. In addition, an optical rotatory dispersion study of the ovaR>tunin conformalional change as a function of dimelhylni/rosamine coo- centration had indicated a concentrationdependent ambivalent effect on pro- tein conformation. Thus, at concentrations of, and greater than, about 200 mM, dimelbylnitrosamine brinp about denaturation. However, a1 concentrr tioro from 200 mM downward the b, constant of ovalbumin increases with the decrease of dimethylnitrosamine concenlratioe, suggesting Ihat, in this oon- eeotratioa ne6e, dimelhylnilroaamine bas a refolding ('Yightenink`) rather thae unfolding (denaturing) effect on the protein eecondary structure. 8ecause of ita protein-denaturing ability, dimethylnitrownine at high concentrations can bring about cell death. In eonlrast, when 20 to 200 mM dimethylnitrosamine is used in esayme and mutagenesis awys, because of its re(olding cffect on protein secondary slructure at thne ooecentrNions, il may produce a1Mtileric enzyme eonformational changes iw vhro, thereby possibly creating artiiactual phenomena. Indeed, the use of nonpharmaeologically high eoncentrations of dinxthylnitroaamine and diethylnhrosamine appears to be the basis of con- flicting reports on the repressibility versus inducibility of dimethylnitrosamine demethylase and diethylnilrosamine declhylase and on the effect of enzyme indu.er prctre.lment on ndro.amine mulagenicily activation by microsnmes. keaenr onvesrrpu.a.s sugaeN rhst the apparent confbct is due to the eslsrence ot h.o rnlymK /orme o/ dinuthylmuosamine demerhylase that rc%porwl in duiuctn,.11r opt.•vu +ass (n.mely, repreasn+n and mdutthun) ts- cnryme in,tu.cr plctrratmrnt Il.r.r ot.xrvahuns and othcrs kad the aulhuu so con- 10 I clude that the cortfentration of dimethylnitrosamine (or any other nilrosamin.) uud in In vitro assays should be kept under 20 mM, and preferably below 5 mM. Since the purpose of studying the melabdism, mutagenicily, and cellu- lar effects of dimelhylnilrosamine and other nilroumina Is principally to pia insight into the mechanism of their carcinogenic eRecl, physiologically realistic testing levels should be observed. Argu., M. F. and Arcof, l. C. Cancer RriearcYt 3t:226-22t1, 1978. From the Seamen's Memorial Reseanch Lboratory, U. S. Public Health Servlo. Ho.pital, and the Department of Medicine, Tulasse Medical Center, New ('rkans. TISSUE AND SUBCELLULAR DISTRIBUTION OF a11-DIOXANC IN THE RAT AND APPARENT LACK OF MICROSOME.CATALYZPd) COVALENT BINDING IN THE TARGET TISSUE Diosane, a commonly used induqrial and laboratory aolvent, lua bees known to eahibil both bxic and carcinogenic activity. As part of a series of investigations aimed at elucidating the mechanism of these eRecN, this paper presents data on the distribution of 01l-dioaane among a number of rat tis- sues and various wbcellular (ractions of rat liver. At various times dter intraperitoneal injection. d'aaane was distributed more or less uniformly among varqus tissues (liver, kidney, spken, hrng, eobn, and skeletal musck), which is consistent with iu polar/nonpolar nature. llowever, in eoMraN so this nearly uniform distribution, uudies of the nature of binding revealed larp tisxre differences. Ttre liver (the main target of careinogenesi.), colon (de- acending segment), and apken showed a greater amount of "eovaknl" binding as measured by Ihe incorporalion of radioactivity Into lipid-free, acid-insoluble materials. Much kss "covalent" binding occurred in the skeletal muscle and blood. Investigations of the aubcellular distribution in Uver Indicated that moN of Ihe radioactivity was in Ihe ey/osol, followed by the microsonul, mito- chondriat, and nuckar /ractions. The binding of dio.ano lo the macromole- cuks in the cytosol was mainly noncovaknl. The percent covalent binding was highest in the nuclear (raclion, followed by milochondrial and mkrosornal fractions and the whole hotnossenale. Pretreatment of rals wilh inducen of mkrosomal mi.ed function olidasa had no aignifkant effect on Ihe covalent binding of diosane Io the various subcellular fractions of Ihe liver. 7lrere is no micrr>M>toetatalyred /w vitro binding of diosane to DNA under conditions in ..hich there is ulensive bindinrof benro(a)pyrene. Woo, Y-T., Arrus, M. F. and Arcos. 1. C. LI/e Sciences 21(10) :1447-1436, 1977. Olhtr support: National Cancer Inslitute. From the Seamen's Memorial Research I aboralory, I1. S. Public Ileaith Senkaa Ilosprul, and Ihe Department of Medi.cine, Tulane Univenily, New (bkaos II

STRUCTIIRAL IDP.NTIFICATION OF p-DIOXANE-2-ONI3 AS 719@ MAJOR URINARY Mt'TABOLITE OF pD1OXANE Intensive diosane eaposure causes severe liver and kidoey daroage and even death. The compound is also a hepatie carcinogen in rata. In 1.lis inves- ligation of the In viro metabolism of diosane, gas chromatography ((iC) of the volatile compounds present In the urine of rats administered dumane re- vesled a maior metaholile. The latter was detectable only at low 1 N kveA, and the amount escreted was bo1A dose- aod /ime-dependent, reachin,: a maai- mum 20 to 28 hours after intraperiloseal intectioo of dioxsne. Administration of diethykne glycol produced the sarns metabdite, but mostly withir, the Brst 16 houn, suggesting that this compound may be an intermediate in the iw viro metaholism of dioaane. Under similar eorditions, ethylene g!ycol, di- glycolic acid and osalic acid did not induce escrelion of the metabolite. The isolated and purifkd melslwlite displayed an intense carbonyl band at 1,750 em-' In the Infrared (IR) spectrum. Its nuclear marnctic resonance (NMR) spectrum indicated two Iripkts and one singlet of equal Intensity at 1.81. 4.42 and 1)7, respectively. Upon GC-mass speclropholomelrie sludiq, there was a parent peak at m/e 102. The nsetabolite wY idenlified u p-0toRSne-2-ooe, as conflrmed by comparison to ita synthetie reference eompound, .rhich ea- hibiled the same IR. NMR, and GC-masa spectra as the unknown. A tenla- live metabolic pathway for diosane is presented; micraaornal mixed function oaidases appear to be involved in this pathway. Preliminary eaperim:nl/ indi- eate that the melaholite is considerably more toxic than the parent compound. Its potential health hazard remains to be asaessed, espeeially since it is used as a commercial and industrial preservative. Woo, Y-T, Arcoi, l C'., Argus. M. F.. Gri/fin, 0 W., and Nishiyama, K. Nounyn :jchmlcdtbrrt i Arrhivct o/ P.rrAoloeY 299.2N )-287, 1977. Other supportt National ('ancer Institute From the Seamen's Memorial Research I ahnrstory, I/ S Public Ilealth Service Ilospilal; the Department of Medicine, Tulsne Universily; and the fkpartment of Chemis/ry. University of New Orkans, New Orleans. EFFI?fT OF MIXED-Fl1N(TION OXIDASE MOI)1f7F.RS ON MNTABO1 ISM AND IOXICI CY U1' 1118 ONC(Xil?N 1)IOXANH The effect of inducers and inhibilon of mised functie.n oxidases (MFO's) on Ihe in riro rnetabolism of dio.anc in the rat was sludicd with both non- radioactive and "('-labekd dioxane. In addition, the effects of various inducen on the acute tosicily of diosane and Its principal urinary molaholite, p diosane- 2-one, were eaamined in an attempt to elucidate the relationship between the mr1alm.lism snd lo.icily of this communly used udvcnt. Resulls showed that prc/rcatment of mate 1praRue 1)awlcy r.ls with the itmhrcen phcrK+barlrild (1'11). h1lyrhln11n.trd h1l1hrnyk /P('111, uN1, to a ntuch ksser eslent, )- I rnelhykholanthrene (MC), increased the metabolite escretion and shortened the lime of onset W peak eacrelion of the metabolile. On the other hand, aa Inhibitor or repressor of MFO's, such as either 2,1-dichloro-6-phenylphenoay- elhylamine or cobapous chloride, decreased Ihe metabolite e.crelion. lbese ra suhs subatantiate the invdvement of MFO's in the /n rlro me/abolism of dioaane. When the relNionship between the metabolism and the acvte toxicity of dioaane was espbred, il was seen that pdiosane-2-ene was considerably more toxic than dioxane. The acute toxicity studies showed an apparent t:orrelation between the metabolism and bsicity of dioxane in PCd- or MG pretrealed rats. However. PB pretreatment had no effect on toxicity In spite of substantially increasing dioaane i melabolism to p-dioaane-2one. Since Ibis lack of PB effect Indicates that p-diosane-2one requires further metabolism to exert ils toxic effecl, the totality of Ihese rewlls auggesls that the generation of the toaic substance from dioxane may Involve a multistep mechanism with pdio.ane-2one serving as an intermediate. Woo, Y-T., Argus, M. P. and Arcos, 1. C. Cancrr Research Je:1621-1623, 1978. Other su pport t National Cancer Institute. From the Seamen: Memorial Research Ldwralory, U. S. Public Neallh Servk. Nospital, and the Department of Medicine, Tulane University Medical Cenler, New Orkan.. RAfNOACTIVE ASSAY FOR ARYI. HYDROCARBON IIYDROXYLASE. IMPROVF.D METIIOD AND BIOLOGICAL IMPORTANCE This report describes an Isotopic AH/I assay, which meawres atl 1he snelahoiites formed during the /n vitro inarhalion of various tissues (liver, in/esline, lung, kidney) with (sN)-ben:o(.)pyrene and accurately determines lower enzymatic activities such as those found in the lung or kidney. TAe addition of Iwo volumes of a IM aqueous KOll/dimenhylsolfoaide (1!/8J; v/v) mixture to the enzymatic incubation medium, makes it possible to aetec- lively extract the unmclabolind benzo(a)pyrene in he.ane. Thus. the radio- activity remaining in the water phase represents all the me/aboti/es synthesized le vitro. While kss sensitive than the more commonly used fluorimelric method. Ihis new Iechnique, whose lower limit of sensitivity is estimated to be about 2.10 'r moks of total metaboliles formcd /ml Incubalhm medium, Is es- Iremely accurale, can measure eaRahepalie and low AIIII aclivi/y, and is not particularly photosensitive. Van Canlforl, 1., De Oraeve. 1. and Glrfrn, !. F,. eiochemtcaJ ond BJoph yrka/ Research Commynications 79( 2): S0S-f 1 Z, 1977. From the I ahoratoire dc ('himie MtdiLale, Inslitul de Pathologie. Sart lilman par 1.it2e. Belarum. 12 1 )

COMPARISON OF ARYL HYDROCARBON HYDROXYLASE INDUCTION IN CULTURED BLOOD LYMPHOCYTF.S AND PULMONARY MACROPHAOES In this study. !n vitro ioducibility of aryl hydrocarbon hydroaylase (AHH) in freshly lavaged pulmonary alveolar aucrophaRea (PAM) was compared to the degree of induction of Atilt in cultured lymphocytes from 13 smokers and t nonsmokers with a.ariety of noooeoplaMie lung disenes. Values obtained for Atilt in PAMs and in lymphocytes from patients withio the rwnsmoker and smoker groups were similar. Levais of AHH in PAMa freshly obtained by lavare were higher for smoken thao for rwnsmoken. With PAMs from sonamokers, bvels of Atilt were virtually Identical for celts cultured for 24 hrs. with so inducer in the medium and for cells freshly lavaged from the lun6 In eontrut, values for PAMs from anakers were reduced to about half the original kvels after culture (or 24 hn. with no inducer in the medium. Atilt values were similar when noninduced enzyme levels were compared for nonsmoker or smoker PAMs and lymphocytes. Also, when PAMs and lympho- cytes from smokers or nonsmokers were cultured In the presence of BA, similar enzyme induction was observed. Results from these studieo indicate that Atilt levels in one cell type can accurately reflect levels measured in the other cell type. Howevet, cigarette smoking dfeets the noninduced as well as the BA-induced levels of the eozyme. This is the first dme that celh (rom two scparsle tissues from Individusl subjecti have been shown to possess similar Atilt Induction capabilities when tested under Identical tulture conditions. By measuring Atilt in two separate tiawes from the same irdividual, probkms previously encountered in using a single tissue for analysis of an individual'a Atilt characterislics might he resolved. This added dimension for the analysis of Atilt in individuals might also provide reaearchers with another tool for evaluating the relationship of Atilt to chemical carcinotenesis. Mcl.emore, T. L., Afarrin, R. R., Toppell, K. L., Busbee, D. L, and Cantrell, 1'. T. The /orrnal of CNnlcaf Inrttrltarlon 60:1017-1021, 1977. Ot6er support: Nationa11ns1itutesof Health. From the Department of Medicine and the Department of Microb:iloty and Immunology. Baylor College of Medicine. Houston; and the Department of BioNogical Sciences. North Tezas Stak Univenity, Denton. ANALYSIS OF ARYI. HYDR(X'ARBON NYDROXYI.ASN A('1'1% 11 Y IN 11l/MAN l.l/N(3 TISSUE. PULMONARY MAC'ROPIIA(iES ANI) B1.CX)1) t.YMPHOCYT['S While variation In the degree of aryl hydrocarbon hydroaylasc (Atlli) inducil+ildy has been observed in pulmonary alveolar macrophases (PAM) and IympMrcylcs lrom diRcrrnl individusls. AHII values in these cclls have aN hrrn relaled w the A1111 s.hvory in other urlulosous titaues In this ahmJy d~r 1111/ ay i..*ty m t.c.h au-tually rrcised Iunif Wiue. Iresh PAM I i I and peripheral bl.od lymphocytes from 14 ciprette smoken (7 with untreated primary lung cancer and 7 eoncancer p.tienu), was measured fluorometrically. Individuals without cancer showed a good correlation (r = 0.973, p<.001) between the PAM's AHH levels and the AIIH inducibility (ezprea.ed as lob induction) in their cultured, mitogen-atirnulated lymphocytea. In the canox patient., however, these values were diaocialed. In addition, the fresh lung tissue Atilt levels and the cultured lymphocytes' fold-inductioo ratioe eorre- lated positively in noncancer patients but oot in the cancer patients. 7lhere was close agreement between fresh lung tissue Atilt and fresh PAM from iodi- vidual cancer-(roe palients, but theae values were only weakly correlated in those with cancer. Upon simuluneous comparison of Atilt activity in fresh PAM and in fresh lung tissue, and AHII inducibility in cultured lymphocytn, an ezcelknt rclalionship was found between these values in all three tissues for individual noncancer paliems (r = 0.987, p<.001). Ilowevcr, thc.e did not correlate in Individual lung cancer patients According to the data, while the capacity for Atilt induction in eaneer-free cigarette smokers is similar in the tiuues qudied. h+nR cancer patients do not have positively correlated Atilt valuea in these tiawes. Mcl.emore, T. L., Afan/n. R. R., Pickard, L. R., Springer. R. R.. Wray, N. P., Toppell, K. L., Matwz, K. L., Guinn, 0. A., Cantrell, @. T., and Busbee, D. L. Cancrr 11(6):2292-2700, 1978. OtAer wpportr National Institutea of Ilealth, National Cancer Institute and the American Cancer Society. From the Deparimenb of Medicine and S1srRery, Baylor College of Medicir and Veterans Adminiuration Ho.pilal, Houston; and the 1)epartmeot of Biological Sciences, North Team State Univenity, Denton. DISSOCIATION BETWEEN ARIL HYDROCARBON IIYDROXYLASH ACTIVITY IN CULTURED PULMONARY MACROPHAGES AND BLOOD LYMPIIOCYTES FROM LUNO CANCER PATIQNTS Lymphocytes and pulmonary alveolar maeropha6es ( PAM ) used foe evaluation of aryl hydrocarbon hydroaylase (AHII) induction were oblalned trorn 13 noncancer and 14 primary lung cancer patients. Fnzyme levels wera mcawred in cells cultured with or withoul the inducer benzanlhraeene (BA). 7Le mean levels of AIIH showed no diQerence belween noncancer and caawr patients for either noninduced or BA-induced cells. Absolute kvels and foid induction of Alll1 in PAM and lymphocyka from Individual noncancer patients were positively correlated. Ilowever, comparison of these values i. PAM and lymphocytes frrom individual lung cancer p.tients demonstrated no positive correlation. In further study, comparison of enzyme activity io macro- phages fr~eahly lavaged from the lung and BA-induced activity in cultured PAM revealed a positive correlation (or both noncancer and lung cancer p.- licnts. Similarly, comparison of eozyme activity in fresh macrophagcs and told induction values in cultured PAM was also well correlated frw M.th groups of patients From these sludies. it would appear that messuiement of Atilt In more than one tissue obtained on the sanme day from a given individual may 1A . IS