Tobacco Documents Online

ANDREWS OFFICE PRODUCTS CAPITOL HEIGHTS, MD (K) s

1977 REPORT oJ THE COUNCIL FOR TOBACCO RESEARCH-U.S.A., Inc. 771E COUNCIL FOR TOBACCO RI•:SEAR(:11-11.S.A., Inc. 110 E..159tb 9ereet, New York, N.Y. 10022

SCIF.N77E1C ADVISORY BOARD to The Council for Tobacco Research-U.S.A.. Inc. as of December 31. 1977 HANS MEIER, D.V.M., Jr. Wg1. Vet., YJ.R.S.11. Senior Staff Scientist The Jackson Laboratory Ba• Harbor, Maine SHELDON C. SOMMERS, M.D.. Chairman Director of Laborato.ies, Lestos HiU Hospital CJinkaf ProJessor of Padwbnr Colk~e of Physicians & Surteooa of Columbia Univenitr New York, New York 1 LEE W. WATTENBERC+, M.D. Pro%ssor of Pothotoq y Department of Laboratory Medicine cac ?'atholoQy University of Minne.Wa Medical School Minneapolis, Minneauta RICHARD M. BINO, M.D. Dbedor of Cwllofo" anI /ntromr.rd Melkine H.mirKton Memorial lloapital, Paaadena, California ProJesaor of Medkine University of Southern California School of Medicine Loa Aateles, Califoroi. JOHN P. WYATT, M.D. Director Tobacco and Health ite.earch lnatitute University of KentuckJ Lexington, Kentucky J05EPH D. FELDMAN. M.D. Head, Department of (mmunopatholo6y Scripps Clinic and Research Foundation La LUa, California WILLiAM U. DARDNER, Pn.D. Sdenrt/ic Dinrro., The Council for Tobacco Research-U.S.A., Inc. E. IC: Hnnt PtiroJessor of Anoronry (erneritrs) Yale University School of Medicine New Haven. Connecticvt Seiewliiie St.R.d TD.e C..wea WILLIAM U. OARDNER, PN.D. Sc/enB/ic Director ROBERT C. HOCKETT, PH. D. ReserrcA Di+rcror DONALD H. FORD. PN.D. VINCENT F. LISANTI, D.M.D. ROBERT 7. HUEBNER, M.D. Associate ReseancAi Dbrrta. Associate Research Director Chief. Laboratory of RNA Tumor Viruses Natbwal Caneer Institute JOIIN 11. KREISIIER, h1.D. DAVID STONE, Pa.D. Betheada, Maryland Associate Researcli Dbector •Assodate Research Director LEON O. JACOBSON. M.D. Dirrcror, The Franklin Mcl.ean Memorial Research Institute RePenuteln Professor of Biolojical Sciences University d Chicaao Chicaao, IUinoia HENRY T. LYNCH. M.D. Pro/essor and Chalnnnr Department of Preventive Medicine and Public Health Crel~ton Univenity School of Medicine Om. a, Nebra.ka

CONTENTS lDttoductbo . . . . . . . . . . . . . . . . . 5 AbturKds d Reporb . . . . . . . . . . . . . . . 6 . Caaoer-Rdaled Studia , . . . . . . . . . . . 6 The Respirator2 Srstes . . . . . . . . . . . - 22 Htart od ChculaNo. . . . . . . . . . . . . . 34 Ncurophumacolop .aid PbyslolM , . . . . . . 44 PRarat.ooiM .nd Bioc6emistq . . . . . . . . . 49 /ereweoio67 and Adapive Mechanisms . . . . . . . 54 p.pideaaiolon . . . . . . . . . . . . . . . 59 MbctUane. . . . . . . . . . . . . . . . 63 ActM Pto)octs . . . . . . . . . . . . . . . . . 64 Ca.apicted no}. . . . . . . . . . . . . . . . . 73 Ldea d rrl.dp.l lavatiplon . . . . . . . . . . . . 83 hde: d Sedor At+lSoea . . . . . . . . . . . . . . M I . Intiraductimn As its quarter-century marh appvachn, The Council for Tobacco Re- search reviews ita history with a.enae of grNiOcatiov over progress in several directiorn, some degree ol frustratba in othns, and a considerable measure of optimism /or the future. In the past 24 yean, TMe Couucil 6r provided substantial support for what may wefl be the world'a most eahriv~e aao-go.ernmcN.l re.scarch pro- pam concerning tob.ooo we a.d healtb. TMe basic policy adopted at the outsrt has been rnaintaintd, that i., so place rNpeMbibility for the direction and guid- antx of the research dfort in ihe Ir.da o/ a Scienli/k Advisory doard, and 1o have the research ee.ducted 67 iw8rpewdeM Investigators in their own insti- tulions. llenee, whalever oD.tribWiar to aciewti/k progress may be attributable to The Council•a program .ri d.. ..ry largely 1o Ihe distinguished men whu have served on this Board. W!M major emphasis .po." cawoer, heart d'aeaes a.d chronic lung ailmenta, the Board has consistently ooe.seped stro.g attention to the multiple steps awd stages in the pathogenesis o1 14u agisell-asaoeialed "eoautitutional diacase.." Basis for Ihis emphasir wp a aou.ictiow tlat rapidly developing ne.w techniques and concepts in biochawolry ahould ptovide powerful tools for elucidatiag these mulliple Meya and ttyer, with a reasonable hope of short-circuiting oL blocking one or more of tlrs aa/ 11ne preventing the disease or delaring its onset. By the end of 1977. tlir prvpr+rtt Aad produced more than 1.500 reports and articks in accredited aKieMi- e iouruaii by scientists acknowledging Council wrpporl. Their ultimate stpiAcaeoe ia relalioe 1o 1he goals cannot be fully appraised at this time. 'Itie rnosak Y eompha and emerging pictures are still far from ckat. 1/ is frequently Ma1ed IW, ItlalorleaNy, many of the most important ad- vances in medicine have 6aw wnda by trpredictable enspiricd diaoovery. Yet. In these daya, such diacoveria ara trtaM likely to be,made by alert Individuals In the course of systanNie.luia Nided by a good rationale. The m.iMenance of scientilic eevitoee+e+w ooad~rdve b t+udr dbooveria is, there/ore. n im- p«t.rw re.po.aiesiitr ot .q tr.Mias oegotiz,tio.. The prerenl report ooal.in+ .bMracts of anauacripts published in 197, qrN acknowledged Cawd spo.roe.hip. TAe aestraets speak for thenmlvea. They illustrate Couecil polic! 1W Sr.Mers alone shall be responsible for d'a- e{oaing their research Andinp i. aootpled medical and scientific journals or before medical and scientific orpwiralion.. The Council is aontinuint Its Oro{ranr of support to iodependeN reaeuce- en in bio-medicine with its original /oal.- , ©

Abstracts of Reports Following are abstracts, appcoved by the aulhors, of reports on new nesearch acknowledging support from The Council that have appeared in tcien- tilk joureals sinoe publication of the 1976 Report. lAe name of the recipient is in italics. The ab.tracts are grouped under these beld'wp: 1. Cancer-Related Studks, 1/. The Respiratory Systen4 Ill. Heart .nd Citcolation, IV. Neuropharmacolop and Physiology, V. Phumacoiop ud Siochetnislry, V1. Immunology and Adaptive Mechanium. V11. Epidesniology, VIII. Macellanoou.. 1. Gwcer-Rel.ted Strdie. Rf'PRESS1eLE AND INDUCIBLE FORMS OP DIMETH YLNfTROSAMINE-DEMETHYLASE Mai. Spndue-Dawley rset and inbred strains of male mice were ustd 1s daM study of diwetb7l.)trwatwlwa (DMN)-0etnetbylase with respect to keetie charockrMka and responae b Ydueer prtlreatmerN; straindependcnce t{ R- ptsibiliy rs1 leducibility was abo inveslipted in the mice. Results d+ow that two eneytne fotse., with diRenea kinetic characleriNics and oppo.ite rrspanses b it dro erapase inducet pretrealment. trder/k hepatic DMN•demNhylaue ec- N.itq. In kinnk sludies, detennination of the Ilo/slee plot of DMNdemr.thyl- tre using a DMN aubatrak concentration range of 0.5 to 200 mM yklds t5ree lsferaoctinR line sepnenb fro+n which widely diReresM K_ and V_ may be calculated. IdeNkail7 panernsed Hofae, plots are obtained with rat and tow.a poeunUoehondrial wperwNaM fractiom, as well as with the ittdated taiao.awtes. The lowiubatr.le-ranps line segment (0-4 mM) and the hish- arbattMe-raa{e aegnre.t (30-200 mM) correspond. in both the rat and the aatw, b two dillerent eatrymNic fornn of DMN-demethylase (DMN-de- metlrYlaaa 1 and 11, respectivelf) which baw (he following diQerent regulatory esarselerittio: (1) PrNreMtwnM of rats and wwoe with the polrchkxinated biplsesyl. Atockor 1251, brinp about represaiow of DMNdemethytase I and it.ductitr• ot DMNderne,brlaia 11• both responses are stronger in the rat Iham in dr /nase. (2) PretreahnerN with phenobatbital trpreascs enz7me I in both pacia• bov.s.er, it induoea eayme 11 only In the rat. ( l) PMzealmerN with )- ntethrkhda.tMer auMtanUallt tepresses DMNdemeth7/ase I in the rat only; It Is i.eQeetivr iu signiAcanelP idluesrind any other enzymatic activity. Impor- tmtly, the genetically disdnct tsalwt of the two formm of hepatic DMNdemi:lhyl- w is suggested by t!w sub.ataslial d+lerenoes in the strain-dependence of the reptrsaibi/ity of enzyme I aad in0ucibility of enzyme 11 In a aeries of nine in- bred strains of mice. The rnara bet ooelaiaa only the inducibk-type of DMN- detncebrlaar kowever, the ranking of inducibilitks in different /rom that of hepatic enrrme H Artau. I C .r .l Z.1rwA.r/r /w R..bh- Arwa rwI AIu.NFAf 89 1e1 199, 1977 Other auppore , - Nation.8 Caeoer In.titule. From the Seamen + Memorial Research Laboratory, U. S. Public 64ealth Service Hospitd, New Orleans, and tbe Department of Medicine, Tulane University Medical Center. New Orkans. ONCO-DEVELOPMENTlLL ALKALINE ?F:QSPHATASE ISOENZYMES Previous studies have showw 1hN fwnor alkaline phosphala,es have their counterparta in relatively early stap of Iwnuw development. For e.ampk, the chorionic phase I boenzyrne in 6-10 week fropboblast mkrovillar tissue con- sists of two heat-acnsilive, L-howqerRinine-irJtibiled b.nds. The fnter one, b.nd A, e.hibib none of LM awa4ewk deterntinants known for vario.n alkaline phosphalase iaoenzymes and Ihe dosrer ons shnea dekrminants of the liver type. After 1en weeas, alkoliaa phoaphat.ae ot the tene placental type appear. in spncytiofrophoblad eelk` and the dwdonlc type beeomes much kss promi- nent. Further evidence prexMad hete Indicalea Ihat the chosionic (6-10 week) tissue iaoent7tne lacks the atMyeak detenrninnNa of term placental isoenzyme. Still more evidence auqeab Maf Ma eoutNerpert of chorionk band A is to be fouid in the fa/it. kadinZ the e.Nwts to questice whether this band is pro- duced by leruocueieoer of dse lestis or by cancer of the lung. In the present study of neoplaslic Ir..dutnnlion in relation b oncodevelopmenlal gene e.- pressiow, ebe tr.cheobroncMd ttett of Mrmana with cancer of the lung wn ehosen w the nrodel, ainw aN tnnMorwntional staaes from basal cell hypcr- plasia 1o earcinorna M aUr nn be eapeeled to be found in the nonlumor areas. Signi/kaM levels of dtdnotwrltYowie antigen (CEA) and human chotionic gonadotropin (HCO) were fowd 1n eatracta of bronchial epithelial cells from twtn•necplaslk as well as eoplrfk ates. It may be, lherefore, that the onoo- developmtntal genes for CEA and HCO we turned on early In transforming cells and persist in malipa.c7. FWlnr.w, W. Il. er d. In: Fishman, W. H. and Sell. S. (eds.): Owco-levrloomenrd CJcne E.prcLion, New York: Academic Presa, Im:. 1976, pp. 163-176. Other suppertr Natiosrl Cancer Institute. From the Tufts Cancer Research CaMer, Tufts Uaivenity School of Medicine. Boston. PIIENOTYPK' ALTF.RATION OF ISOENZYME PRO1'ILFS OF Al KAI.INE PIIUSPIIATASE IN HeLa TCRC-I CELLS GROWINU IN IMMUNOSUPPRESSED RATS When grown in culture, the human cell line, HeLa TCRC-/, produces the heat-stable eareinoplacesNal Repn isoenzyme of alkaline phosphatase e.clu- sively. However, as Ihis paper describes for the Ilrst time, a prafound alteration lakes pl.ce in Ihe isoenzyme prolYe of alkaline phasphalase when 11c1 a I is grown as a solid n.mw in hnmunoauppreased rats. Acrylamrde AeI eleclro- phoresis reveals a decrease In the Regan Isoenzyme with a somuhanahr, apprar- ance and increase In new fasl-moving iwenzyme buxls. After 70 days of 7 6

growth lw .iro, no Regan isoenzyme Is visible and a singk fast-moving iso- enzyme band is Ihe dominant isoenzyme. Tl+is new enzyme form is identifkd r the onrnamnionic (FL) isoenzyme lint characterized in the FL amnion cell line and later In a hepatoma p.tient. Upon transferring the tumon back to cul- lure, the oefls regain their original Regan phenotype after one to two weeks. Ne.erthcka, even though the esprrs+ion of the Regan isoensyme is restored, the NeLs TCRC-1 cells' isoenzyme eapression has been changed, since lhis cell line after being passed through an irnnwrwsuppresxd rat Is able to rerxpresa the oeeo.mniotie isoenzyme N certain cell demities in culture. Such growth of humaw eawa cells in inuswnauppressed snimal hosta may provide a model trystem for the study of carci.ogenic espns.iow. Sioaer, R. M. and Fisllw..w, W. H. le: Fir6man. W. 11. nnd SeB, S. (eds.): OwcodrrrloPm.nral Grnr E.orrsslon, New York: Academic Presk l.c., 1976, pp. 177-161. OtArr sr.trorfr National Cawctt Institute. Frvm the Tufts Caoeer Research Cenler, Tufts University School of Medicine, 11041100, CARCINOPLUCENTAL AI.KALINE PIIOSPHATASE: BASE LEVEL AND HORMONE-INDUCED ACTIVITY ASSOCIATED WITH EVENTS IN THE CFLL CYCLE The Regan isoenryme (a caminoplacentsl Senc product flrst discovered In a brvnehornie cancer and subseQuently found in a variety of other human eanecrs) is a placenld alkaline phosphatase isoenzyme which is also produced by the NeLa cell model system. IIeLa TCRC-1. a particular cell line which h wrorwphenotypic with respect to the Regan Isoenzyme, is esamined in this study of predni.olone regulation of alkaline plwsphatase as a function of events In the ceM cycle. Results show that DNA synthesis is not required for horewne {nduNion of the Regan isoewzyme since induction occurs in the presence of hydtoayurea, a specific inhibitor of DNA synthcsi. AddilionaMy, when partial- ly ttqnchroniud cells are dlowed to leave the S period prior to hortnone treat- ment, snd hydroayurca in sdded to prevent cells from entering the neal S periiod, hotnwne induclion of the Regan isoenzyme is stip observed. This indi- calea that Initiation of capression of hormone-induced csrcinoplscental alkaline phosphalase occurs prior to the DNA synthetic phase of the cell cycle. This Mding can be harmonized with previous wudies on other cell linn by hy/dhe- sizing a Iwo-skp mechanism of bormare actioA in which the Ant Induction step reauins DNA synthesis, while the second Mep, which is more closely rr.lsled to transcription and translation sequences In the Or period, does not. It Is this secord step which is functional In the TCRC-l cells. SinRr, R. M. and FLhmaw. IY. H. DI//trtnriarlow 3:127-I )2, 1976. Other .. pr.r1 r National Cawur Institute. From the Tufts Camcer Resesreh Center. Boston. t DEVELOPMErTAL PHASESPECIFIC Al.KA1.INE PIIOSPIIATASp. ISOENZYMES OF HUMAN PLACENT.4 AND TIIEIlR OCCURRENCE IN HUMAN CANCER The appearance In ttrnors of a devebpmental protein antigen such as Regan isoenzyme, a ploce®ta1-typt: alkaline plqsphatase, may indicate 1he re- eapression of a gene set eNaraNeristie of a particular event in early develop- ment. Information of this aort may help elsasify slages in neoplastie progres- sion. 'TTThis preliminary eomtnwtieallon descs:bes alkaline phospha/ase ekclro- phordic patterns char.ctcrislie of three plsses in early human trophoblrtic development. The Phase C(6 b 10 weeks) pattern consists entirely of two heat-sensilive. L.homarginise-Mhibised bands, the slower one possessing anti- genic determinants of the liver-bone type of alkaline phosphatase and the faster one lacking any of /hs knoww alkaline phosphatase antigenic delermi- nants. The Phase 2(11 M 1) weeks) pattern shows a miature of Phase I and Phase ) (11 to 16 weeks) boewttynN oowtponents, the Islter of which has two boenzyme bands with the cA..ticUalslics of /etm placental alkaline phosphatase. These lhree pattetas of d t.elopmdrty phse-speciM1c placental alkaline plwse phatse corsespond, in order a appesrsnoe, lo non-Repn isoenzyme, a miature of non-Repn and Reps btoewiywas sed Repw tsoenzyme as are found In a variety of human cancer tlraines. Etswtpks of each phase's occurrence In humas cancen are given, and Iw thig way s guide is provided for interpnNing tumor alkaline phosphatases tr reBeeliowa of the activation of phne-speciQe tropbo- blaslie genes. Fehman, L., Miyayama, H., DeisooM, 10. and Flsbmaw. IY. H. Cwcer ReuarcA 76:226fi-2272. 1976. Other w'prtt Natiot.d Cawoer /sntitule. From the Tufts Research L1etMer and the Depatmeia of PatholoRy, Tufts Uni- versity School of Medkiwe: the DeparlmnN of Pathologr, Ilarvard Medical Schod, and Boston Noapital(ar Wosner. Boston. REGULATORY CONTROLS OF ONCOTROPIIOBI.AST PROIFINS ANI) DIiVELOPMF.NiAL ALKALINE PIIOSPNATASFS IN CANCER CI?LIS Regan isoenzyme (plaurttal-fpprr I/kdirre phosphatase) and human chori- onic gonadotropin (IICO) are two onco/rophoblasl proteins which ate onco- developnrcntal gene products readily studied'in human cancer patients and in several eaperimental sTslema. Three such models are: (1) llel.a suhlines TCRC-l and TCRC-2, which produce Regan and non-Resan iaoenzymes, (2) Hep-2 and FL anwJow eeB lines acting as models for the reciprocal espressiaa o( de.elopmenlal genet, and (3) /w dw moduluion of developmental gene c.- pression M Ilel.a cells. in this la.l Indanee, for etanrple Hel a 7CR('-1 cells gro.w in Mnmunowpp.essed nls, fonning a tunwr noduk which eap.cssn a nee oncoanwJon (FI-) i.oeetlnw. whila the Repn isoenzyme disappcns. When these tumor cells an returned so eullure medium, however, the FL 9 g

species d'wppe.rs and the Regan isoen:qme reappears. This particular m)del ir expected to prove very useful in interpreting tbe varioun data obtained esith oell cultures and tumors from cancer patients. A chronology of early develop- sneet hr already been mo.t helpful in this reptd, since the counterparts of a number of lumor proteins appear as earl7 as 6amelo6enesis and as late as ten weeb of gestatbo. Flthw..ti W. H. and Singer. R. M. C.wee. Reu.rA )6:12564261, 1976. OrA., a.pr.ret National Cancer I..tihNe. . Praw th. Tu(ts Cancer Research Center. Tufts Universily School of Medicine, Bouu., ..d the Department of Anatomy. Fairkigh Dickinson University School d DeMbtry. Hacltensack, N. 1. EVIDENCE FOR ALTERED OEN13 RE(lU1.ATION IN HelA CELIS RETURNED TO CULTURE AFTER OROWINO 1N IMMUNO6UPPRtSSEi) RATS A pro(outd change of phenoypie espression oecun when Hel.a cells cloned for the exclusive production of the Regan isoen:ryme of alkaline pboa- 'hatw ara Rrown in imnwnosuppressed rab. In the course of a sequential al- lerstione i. 1he i.oenaryme eapres.ion during tumor growth, the Regan isoen- t7rm dia.ppean and ae osooannioa (FL) isoeas7me baomes the domin.t (orns. Tb Preaest report deseribes the isoen:yme reRulation after 1Aese cells rv teturned b culture (or at kaN three months. Thew celb In cuhure demon- atraM a detrity-dcpenden( aheratio. 1s boeruyme proflb. The ReRan i.o- enz7wu Y the dominant itwewsynw (ons in eew. sparsely populated cultures. ..hile dn o.oo.mnion iweat7wu predonan.ter in the later hi6h-desitp sta=e" o( po.lh. Such a eeM Nan wMch w be made to produce the Regan isoen- s7t.. by etilerritrR cells In low deniry asd whkh wiN re<spran the oaotrm- .ioa (PL) ioeezyme at hi6h-veM den.iy might be very useful in the study of Molat:r.n re6trlatioa in hwuas eaeoer cells. The eBects of pr~edniso(one on 14ne odlo ia e>r/1wu are variable, depending on the time of ib addition. A tnarted i.e.ear in artrme k.eb Is seen whes the hormone is added early in thw pawftr eTde. P,aa7rns bductioo b restricted to the Rep. ieocn:yme, whUe hormoae-tnedlaled d'ani.utio. M ars7me activity is confined to the oncoamnioa (PI-) Moebs7wn- Sieller, R. M. (F1.A.n.% W. N.) C.wrer Re.r.rA )6•1262-1265, 1176. OfA.r wsrr.rer National Canoer Institute. Prem the Department of Anatownr. Falrki6h Dickinson Uoivenit7 Scbad of Deotislry, Il.ckensact, N. 1. IDENTIFICATICN OF A DUTA4tiOL-EXTRACTABI-E HUMAN PLACENTA-SPECIFIC ANfIt3EN WITH ALKALINE PHOS/•IIATASE ACIIVI-IY This study focuses on the placental meerbrane-a+sociated antigens released by n-butanot, an organic sd.:ut, whieh wlubilizes a number of cell membrane components. Most of the tisax Bpid L extracted into the alcohol Iraction leav- ing the gtycoproleios, polysaocharidea and other watcr-solubk eomponenls in the aqueous phase. In this seia of experimrt,;-. bomoRenized whok-term pla- centae from different individrah were tteated c-ith n-butanol and the immuno- gens-conlaining aqueous (racUons were used w rwe hetero(olous hyperimmune .era in rabbits. ImnNrrwekcarophoresis of the anti-placenta antisera tevealed at kast sia aolillenic coanpoeeeb in the placental eatracts even ailer they had been completely absorbed with pooled male serum proteins. I(owever, the anti- sera so absorbed, designNed (-PMS). still reacted strongly with estracts of normal adult intestine and Lid.e7. Similar reaction patterns were also obtained with individual bulaaot eatr" of I I various other normal adult tissues and with a composite extract ooa/aiMitd equal amounts of each of the 1) individual ealrads. Of the sia antizenk oorapoaewtu in tbe placental eatracts reacting with the (-PMS) anti+era, only Ilr oae axoci.led with alkaline phosphatase activity retained its reactivilr. Homoje.eors placental alkaline pbosphatase contlrmed this i.nnwno{o1ic and enzymologie Identity. F.atracts ftom each of three pla- centae injected in1. /Moe pain of rabbils aR produced an identical antibody reaction with the unique delenni.anl(s) of plaocMal alkaline phosphatase. Ea- tracta of 14 olher placeMae re.cled with each of lhese antisera to form a single precipitht line of identit7. These rewdtr Rrnttr establish that placental alkaline phosphatase is a characteristic Plaonu-speeiAt (eW proteio. ChaoR, C-H. and AnRe1llo, D. (FLkrsara,l6'. R.) The lewwd oJ /wrnrwolop 117(1): f1-16. N76. Other atarr.rf r NNio.al Cancer IrlitWe. From the Tu(ts Research Cewler .rd the Department of Patholopr, Tufts Uoi- •enity School of Medicim SoMoa I DIRECT IMMUNOPEROXIDASE STAININO FOR REOAN ISOENZYME OF ALKAUNE PHOSPHATASB IN HUMAN TUMOR TISSUES ImmunohiMochemicd loeatitatiow of Regan hoen:yme by a direct perosi- dase-IabelinR technique wr obset.ed 1s eenain selected cancer tiswe cells by both light and electron microaoopy. Among Ove tumors selected for e.amina- lion, two cases (I & 2) that were hiNOChemkally confkmed by their e.lreme sensitivity to L-phesylatanirr (LPA) alm showed a specific immunoperoaidax reaction lor Regan isoesut/me and did not show any difference between rhe histochemk.l Iocalitaliom of Regan hoearyme and the immunoperoaidase stain- ing one at the light micro.oopy k.el. The Regan isoeniyme in Ihe.e cancer utls was seen predominantly in the eytoplasm containing diffuse and fine granu- /ar reaction producb, but it was aHo noled In the cell border area. Immuno- ertochemical studks at the eketroa microscopic level were confined to the lumor tissue from case I wlKre Repn boeneyme eouW be specifically demon- tlrated on the plasma membane. On the other hand, cancer utls /rom case 3 II • 10

containing L-hornwrginine-aensitive alkaline phoiphatase isoenryme acd from cases 4 and S, selected because of the absence of alkaline phosph.tase sclivity, wen: apparently wtrcaclive to the specific immunoperosidsx staining. Based on these Ilndinp, peroxid.ae-labekd antibody specific for human pl.cenl has been wocesfuliy used for deteeting Regan hoen:yme in cancer cells, because the identity of pfacental alkaline phosphatase with Regan iooenzyme kas been weR established. The authors feel that theae LPA-sensitive lumors represent a dr of cancen that should be lsveatipted from the point of view of esprea- aiom of embtyoeic genes. • M67ayasu. H., Doeltpst, O. 1., Mensoll. V.. Oandbhir, L. and Fb6rnan. W. H. Cwcer 3t(J):12J7-1216, 1976. OfAer arrpwtr National Cancer Institute. Prorto the Tdta Canoer Research Center and the fyepartnsent of Pa ihok.gy, TtJln Usi..r.ity School of Medicine, sabs; and the Ikpartment of Pa holoBy, Powd.ak. Hospital. Wdpoie. Mr. TRACHFASRONCIIIAL EPfiHF.1.lAL MULTINUCLEATION, VIRAL INCLUSION sODIFS AND MALIGNANT DISEAS6 This epidemiolngicd study was undertaken b determine asy possibk re- laliorrhipa between trachcobronchial epithelial multinuckNion. viral inchniw+ bodies„ and malignant disease. (It is already knows that patients with malig- sant di.ease have an increased incidence of multinuckation and thal there is a aaaonal variation in the number of virat inclusion bodies in the ciliated epNheliwn.) For this study, smears from /,130 patients - of whom 1.024 (V vuip A)'had various malignancies while the other 3.126 (groups S and C) wern sufferisB from many types of norunaliBnant conditions - were esamincd for 1he prvsenoe of viral inclusions and multimakaled ciliated epithelial cells. Low deped of multinuclealion were observed nwsl frequently in 1he wmmer is p.tkatM both with and without known malignancy. A6o, cylopiasmic indu- abs bodies were aeen least frequently M the summer .nd auturnn in all patients. Wbe. &.on, A and group C were eompued. siBnit{cawtly rsom patients with canoer rad ritsl 4nausion bodies. Th. aeawnal ineidence ot hiBh pt:roentaBes of swllisrckaled cells in patknts with viral incMeiona bst without known cancer (V oup E) followed the general pattern of being lowest In Ihe summer; but in t)wat patients with cancer (group D). peroeMales of nwltbwck.led cells were hlBher f. Ms summer moMAs than in any of dre other aerorn. Ttrese observe- tbae h.ve led Ihe authors so conclude MM IrachcobroncAlal epilhelW rwuNi- swckatios scema 1o be triucred (1) by malignant disease anywhere in rhe body osd (2) by the seronal prevalence of respiratory viruses. Most important- Fj, this wort indicata that respiratory viruses may have a apeciflc eRed on the ciliated epithelium of casoer patients. CAdow, 1. er .f. TAe /e.rw.t o/ RrP.wdrnlve M./tcfwr I f( J):111-/46, 1477. From Ui. Department of AeathesioloRy, New York Univenity Medical Center. N.w York. I ENDOOENOUS C-TYPE VeaUSEl: DOQ:OLB AGE!NTS IN NATURAL L9: H PROC.'SEt About 20 different varftbra1ft specks .re no.w known to harbor C-type kNA •:iruses. In nnn, 1hi ptaeao. of tbese particks is suggested by ekctron microacopic, biochemkal .ed seisloBic atudie, although  deflnite replicating human virus hq yet 10 ~e hou*ed. 7Lese endoBenow viruses are inheritcd through the term ceM anC Ibeir production is regulated by genetic information carried in the host cell. Tro classes of esdc~,~esaus C4yp* viru.ea, «o(ropic and :enotropic, which may hi vr Iheir eowNerp.ry In nwa, have been rrcoBnirtd In certain animab, putka/utl the srouse. Ecotropic vinres spread through the host, can be easily lnsamitled to alla of the sune species and can induce malignancy. Xenovopie .hraer e.nnot isfed cells from their host species but are Infectious for cells of beNrdoRosr apecies. The interactbn between these two types of virv.es eoW tea.N in the traw.fer among apecka of genetic in- formation relaling to worraal site pvoea.ea and malignancy. lbe auUwr eo.- chrdes that endoBenow C-t7pa vinrea may be positive regulators of embryo- genesis. differentiation usC normal cell devdopmetN, as wep as of the e.tremu of ehese prooesaea, eeR a=irs, awloierw.a diacse and cancer. L.4vy, /. A. tiornedkL.e 21(2):i4-fJ, 1976. OtRer a.rr.etr National Ctno.r IsrlipNs. Protn the Department of ALdidM and the Canar Resc.rcb Irotitu/e, Univer- aitr of California School ef Medicist+, Sas Francisco. ENDOOENOUS C-lYPB VIRUSFS IN NORMAL AND "ABNORMAL' CELL. DBVp.IOPMHNT In this aomprebensi.e e:asAsalios of ewdogenoua C-type viruses In nor- mal ud "absormal" c,er ievelo}serK, tlr vhwes' potential ro{e in the proc- esaes of normal swrrNio. (embryornesia sd diAerestiation) and aging (au- toimmunitr and eanoer) an discuwed. These Gype v'iroses, which are budding RNA viruses with lipoptsteiw ot»b, an endogenous, l.e., inherited in /he lienes of many specka, aed have been bwp/k.1eA In cancer in several species of animals. Study of the role of thex dnres in rAum has been aided greatly by the mouae nwdel aystem, since s.ioe develop dbease. similar to tlase of humans. Thnee classes of mwiee C-typa vMuses have beaw idenlifkkd by host range snd sero- bBical studies. Owa of these dres, eaemplilied by the aenatropic virw. M present is early embryoo, in cells undergoing normal differentiation and is cancer cells. The other daaasr. eoouopk; and amphotroyic, ue found primarily in animab with pathological - owditioa+. AN drest C-type virusea are eapressed apoManeotrl7 by oeN. N eertain frequenciea'and titen during the litetime of the mouse. A aonisnwnoBiobrds fac/or Iw been identifkd in normal mouse aerm which specifkaQy weutnliaw IM aenoUopie virus class o/ endeKenous C- type viruses. Virw eapres.lon, therefore, appears 1o be rcpdated by intr.ccllu- Iar r well r humoral faclon. These observations suggest that C•tyl.e vwu.ea play a rok in natural procere, particularll normal matwation aed aging It 12 13

Y also suggested that these murine viruses may have counterparts in humsns in wboee tissue eedo=enous vitvs particks have been detected. Le.y, l. A. C.wcer Rex.nrA )7( s ) :2957-2%!, 1977. Other aorprtr National Cancer Institute. Frar the Cancer Research Institute ..d the Departtnent of Medicine. Univer- sity of California Medical CeMer, San Fra.ciaoo. ANTIOEN-SPECIFIC NONIMMUNOOLOBUI-IN FACTOR THAT NEUTRALIZES XF.NOTROPIC VIRUS 13 ASS()CIAIED WITH MOUSE SERUM LIPOPROTEINS (MURINE C-TYPE VIRUS/ ULTRACENTRIFUOAL SEPARATION) Mouae sera sped/kdf7 acutnNoe tunottopit: but not ecotropk virusts, this netrtraNsabu being atlribMable b sotne solubla aerwe factor and not eflected by insmutwklobulins. Secwre both aenotropk vinn and the soluble neoltc lizinB factor (NP) are present in a11 ho~ne mioe. the suthors have suggested lluit NP plays a role iw the regulation of endogenous ><enovopie viruses. Interaction of NF with the virus N the cell surface may change the intereal mika and lhere- by affect normal life proeeses. This serum factor is furthet characteriusd In d+sa report. It is slable at pH 2-7 and mists ether ealraction, freezing or brief healing to 100' C. Ultracentrifugal separation of immunoglobulins and lipn- prolein dernonstrates Ihe association of NF wish the serum Gpoproleim. Oel perwreaiion chromatography tocalires it in the region of lipoproleins and IsM eaplaining the previous conclusion of others that it is an immun"lobulin. Vari- ae procedures also show that it is not the 70.000 molecular weight Nlycopro- Iei.(s) dreubtinN in maae serum in high titer. UltrscenlrifuNal fractionation of twaw Npoprolek+n Indicates that most of the NP activity is concentrated ie 16e density M/erval 1.019-1.175 d/cros. Additional studies alwuld determine whether NP is associated with oee of the major da»es of lipoproteins or with a spetiNe tei.ur Iipoprolelw specks within this density interval. Preliminary evidewoe sugSens that a protein srokty of NP may be important for neulraiiza- tioa l.,o.w density lipoproleiaa in human serum have beew teporled to have a regulatory effect on celt-mediated hnnNwnily, blocking thymraderived (T) ceM activatiow and rnited tymphocyle reactions. The reoo~nition, reported here, of a factor associated with mare serum lipoprdeins which specifically Inactivatea a virw represeuts a nonirtwnunoklobulis neutralizing reaction to an endokaaua virus by a hoN. Laon& 1. C., Kant,1. P.. Ohsdo, O. and [.ery, l. A. rrocerl/ngs of the N.Nond Acalawry of Sciences of tAe Unlrrd Stars of Amr.- /c. 'f1:276-2t10, 1977. Other nw'r.rfr National Cancer Institute and National Heart and Lung Institute. Pron+ the ('ancer RexaMh Inslihote, the Cardiovascular Research Irntitule and the Departmenl of Medicine. Univenity of California. San Francisco. I INDUCTION BY CfOARET'iE SMOKE OF ARYL HYDROCARBON IIYDROXYLASE ACT1V[TY IN THE RAT KIDNEY AND LUNG Previous Investigators have reported a relationship between the inducihilil7 of aryl hydrocarbon hydroxylase (AHH) activity and wsceplibility to the de- vebpment of skin lumon in mice ud possibly IunR cancer in man. Thn study of ciearette-srnoke-Indueed AHN activity represents an attempt to understand the biochemical reasons for sudt a Nnkate. A 1S-minult etpowre tq a 1/13 dilution of cigarette smoke /oduoed ANH in lungs and kidneys from Sprakue-Dawky rata. la bcth hseR and kidney there was a 2-how lag betwee. smoke Inhalation and the enayme isduMo. on.et. Thereafter, the Ali/l activity Increased very rapidly to atak about two hours laler, reachinN a muimal ac- tivity which corresponded itv thtea or fow limes that of normal rats. Up to four successive inhalalioro admt slqtted at 2-how Intervals induced both lung and kidney h7droaylase activilt s c.dditlvell. The matimal eQec1 corresponded to ah rN 10 timn the coMro( vahr. Compared to the kidne7 eezyme, the hwk A/1N activity was about Ihae er four tMres mae sensitive to small coocentra- lions of cigarette smoke. h(enrresweet of RNA and proleis synthesis require- teeMs showed that protein oyahesia was continuously required for the eslab- IishmeM of the AHH Iaductian but RNA synthesis was only necessary in the Initial period of the pheooemeost. To further characterize the lunt and kidney AHH activiry, the half-life of the eigatetle-smoke-induced AHN activity was compared to melhy{cholaMbrewe-Ipdueed AHH activity. Overall, the authors feel 1ha1 their results suppert the hypothesis that the inducing agents in cip- MIe smoke might not be paiyc7dk hydtourbons. Van Canlfott, ). and Cki'ew, l. IntcrnsHonaf lorrnd of Cwcer 19:S7R-SIS, 1977. OtArr w'portt Foed National d1a la Reclterehe Sciemiflque. From the Laboratoire de CrYeis Mbdicak, Institut de PatholoNie, 1.fte. Bel- gium. ARYL-NYDROCARSON HYDROXYLASE ACTiVITY IN LYMPHOCYTES FROM LUNO CANCER rAT1ENTS AND NORMAL CONTROLS • The aryl-hydrocarbon hydrvayla.. (AHN) system, which metabolires and Is induced by a wide variety of omepoundti has been shown to be genetically determined in oertaln strains of mioe. It hu aho been suggested that the level of enzyme irducibility In human lymphocytes Is genetkally regulated. In this qudy, a new technique involving r.diomNrie assay was used to compare kvels of A1111 activity In IymphocTles of hng canoer palknts and normal controls. Subjects Included 11 mde patients with histologically verifled lung cancer (nine squamous cell eareinoanas end two adenoeareinomas) and 1I age- and sea-mNChed controb. The luns cancer palknb e.hibiled considerably more variation in Atlll activity than the controls. In addition, the mean Alllt ac- tivity of the hmg cancer patients wr nearty four /imes that of the controls. Similar rrsu/ts were obtained from comparisons made between the eight ciga- rette smokers in the control lroop and Ibe aNe-matched lung caneer patieMs. The cancer patients exhibited much more variability and a significantly higher 14 15

mean level of Atilt activity than the control group smokers. While discussing the implications of this work (or lung cancer epidemiolo6y, the authon suggest that AHH, by virtue of its possibk genetic polymorphic status coupled with its ability to mctabolire certain chemical carcinogens, may be of estreme value as a marker for delection of cancer susceptibility. Ouirtis. H. A.. LynrA, H. T., Mate. T.. Hvris, R. E., Wells, 1., Caha, L., An- derson, l., Mdoner. K.. and Rankin, L Oncoio" 33:105-109. 1976. . Oth.r .nrP.rt t National lnatkub of Health. From the Department of Preventive Medicine. Crtighlon Uni.eniq, Omaha. ARYL )IYDROCARBON HYDROXYLASE ACTIVITY IN PUI.MON,\RY MACROPHAGES AND LYMPHOCYTES FROM LUNG CANCER AND NONCANCER PATIENTS Pre.ious reports have wrMested that aryl hydrocarbon hYdroa7la.c (.iHH) Inducibility may be associated with the propensity to develop lung catrxr. In order to shsdy lhis hypolhesirted relatiornhip. AHH activity was meawred In pulmonary alveolar macrophales ( PAM ) and peripheral blood lymphocyles /rom 47 patients with primary lung cancer and 56 patien/s bronchoscoled for a variety of other medical problems In PAM (rom nonsmoking patientn with- out cancer. the Atilt level was 16±2 milliunils/1(N ce®s, whereas noantnoken with primary lung cancer had values of 24t3 mdliunils/ 10a cells. Ci;arHle srrakers eonsistenlly had higher PAM enzyme vahres lhan nommoken, al- Ihouglt srtwken without cancer and smoken with lung cancer had simi'ar en- ryms ktvels. Induction of Atilt in cultured lymphocytes (torn nrn+unoken was signi/kantly lower for lung cancer patients than for noncancer patients. Enzyme indudioe wa similar in lymphocytes from smokers whether or not tln:y had hsng eancer. /mportantly, with cells (torn Individual petienls without lung cancer, a positive correlation between inducibility of AHH le cultured lympho- eY1es rrd AHN activity ia PAM was noted. On the other hand, with ce11, /rom patieau wilh primary lung eancer. PAM and lymphocyte .ahtes were not poei- Iively oorxlated. Further investiptions will be rKcesaary, to delineate the rnecha- nMtrr responsible for Ihe dissociation between AHN .ahtes In PAM and those in lymphocr/es from lung eancer patienb. Howeoer, for whatever reasons this phenomenon esists, k may be of diagnostic vdue In the early delection of lung cancer. McLee+ors. T. 1.., M«An, R. R.. Busbee, D. L. Richie, R. C.. Springer. R. R., Toppell. K. L, and C.Mre11. E. T. C.tsrer Reu.rcA 37(d):117s-11t11, 1977. OfRir wPP.rft Nallonal lnatitutes of Health, National Cancer Institute and the American Cancer Society. Pmm the ikpartme-ni of Medirine. Haflnr College of Medicine. Hon.Mn. and Ik1..rimeM• nf 11.,M,fa.l S[Kn.r. and h..ic 1ledlh lcknce.. Nurth iexas State l lm•rr.1i1 sn•/ o/w / r..t t nIM fe ../ (1.a+.paihM Medicine, Iknlon I IN VITRO INQUCTION OF ARY[. HY'DQOCARBON HYDROXYI.ASE IN HUMAN PULMONARY ALVEOLAR MACROPHF.GES BY BENZ.ANTHRACENE Ttn polycyclic hydrocarbon, beannlhr.cene (BA), is capable of inducing aryl hydrocarbon hydrotylaae (AHH) in human pulmonary alveolar macro- phaRes (PAMs). In the a7udy presented hne, time and dose-response curves for masimum in ritro inCudion of the AHN enzyme system in PAMs were established. In addition, a positive carrclNion was demonstrated between the kvel of AHH activity in E'AMs freshly k•:tVd from the lun6 and the degree of induction of AIIH in cultured PAMs from individual smokers and non- smoken. When PAMs fr+arn either smokers or nonsmokers were cultured for 24 hours in the presenoe of 10 rM o1 BA per vial, si6nific.nt induction of Alill occurred. indueed vahres o: Ai1H were over three times those of non- Induced .ahres for PAMs frowr tnwoken, while induced Atilt values In PAMs from nonsmokers were o.%r iusr lima 6reater than the rwninduced vahtcs for the same indi.iduah, indietlMt; tdM BA is s good inducer of the Atilt system in cultured PAMs. Measuc twesats sach r dre+e of the inducibilitr of Atilt in cultured human PAMs cae)d povide aw eaperimenlal system suitable for study- in6 the mechaniues respa abM for the idtiatiow o1 pulmonary carcinogenais. McLemore, T. L and Ma/n. R. R. C.nrtr Letters 2:327-333, 1977. Other ..rprtt NationallsmMula of Health. From the Departmenb of Medicine, and Microbiology and Immunolopr, Baybr College of Medicine, Houstoa RELATIONSHIP BETWEEN LEVELS OP ARYL HYDROCARBON HYDROXYLASE ACTIVITY AND SU9CEPTIBILfTY TO 3-METHYLCIiO1.ANTHRENE AND BENZOIQ)PYRENE-INDUCED CANCERS IN INBRED STRAINS OF MICE Susceptibility to 3-metbrktholanthrene (MCA)- and benro(elplrene (BP)- induced fibrosarcanas (and eareiwama) Is linkedd to naturally occurring diRer- ences in aryl hydrocarbon hydroaylase (AHII) kveh, that is, where Atilt in- ductibilit7 se6regales a a oodowtisrnt 6ene, or wherj noninducibilitr is domi- nant. Moreover, under condilittnI wht:re AHH h:vel: are arti/kiallv altered, ar occurs after trealmenl with the chemical defoliant 2,),7,8-letrachkwodihenro(sl diosin (TCDD), 1ha Inherent tesiManae of AIIII nonresponsive mice to MCA- inducrd tumon can be o+ereome. Moux genetic model systems show that when A/1H responsivenees to MCA treatment sepeples as a single autosomal dominant dene, either a sirt{le aMOawnal codominant gene or rw+nirducibi/Nr can be dominant depending on /he alr.in. of mice employed. In each of the Ihree genetic regulatory systenr described here. the level of Atilt inducibility is correlated (or linked) to wncrptiMli/y lo lumor induerion by M('A. Nthete AIIII Inducibility searegates as a single dominanl gene. su.cepubilay to M('A- induced lihrosarcoma h correlated with esprevfion of /hat jlrne function Susceplibilily to BP also oorrelales with high levels of Atilt induciAility. I/igh doses of TCDD given It hours pior to MCA result in tumor incidences simi- 16 17