Tobacco Documents Online

48 CTP HN 0 11 B'72

I i . EXTRA COPY REPORT O, THE COUNCIL FOR TOBACCO RESEARCH-U.S.A., Inc. 1973 ~~ ;t• ~ ~ {.~ ,, Mv~•I•. a I . 11 o I .C1 .) ~;~ ~~~• ~. ,~• f

••4 Organization and Policy 'Tlw C.ouncY /or Tob.oco Rcnearcb-US.A., loc. ..s fortuoate this ycu I. h..1.g ..rorld•nsowoed asoer r.e.Uplor. Ck. WiUl.re U. G.rdeer. /oi. th+ u.A as 3ciesti6o Director. T'iti4 p.t W!.r ..cam .ioea the deah of ~' Dr. CUrcno. Cook 1Jtt1. i. DKC+..rMr 1971. Dr. Oardner receatty reNred .. • E. X. HunM rrof.uw of A.Noery at Y.Y U.Iwalf7 School of MedkMe what ` b bad beer hdmor ad Chal.>..a ot t!o Dqortr.e.t of Ae.ro.n7 /rom 1967 191) . - ~, Or. Robert C. Hocteft, w•io J~.m brw rk! Tbs Cowc* d.oe 1931• S.t b..e ypoLM.f Rewre! DMstor. 11o W iw artil.t r AetZ n6 Sckotibo ~' Weasar for tfr /a.t two 7s.m T1.o dhtl.wiJd aM." /olr/ the Sd..df{c Ad.br7 ro.rd drrleg 197). iddYi6 (M1Asr areyt6 ..041 oaprli- b 1y >>bard. Tlwy .rt Dr. Averill ~~ ~/.. p. U.6o.. hofwor md C7dr.rm ot 1i. D'.rt.w of ht`oio6y N tL. U.fw.N7 d C.Yfor.i. Scfooi .t if.delr i. Sr Dieoo4 ..d Dr. Heory T. Lys* 1'roforor ud C1.in.r of the Dep.eu.eM of t•r..wl.. M.dki.* ud , • lv6Ye H..kl .t Cni6foo. Ud..n1t7 lebol of M.dki.. I.Os.ha. Neats.i.. ..- /' tli be l ft ttr rao d McKeca . tA 3r n . r . . 1.+r. e u >L ~; . C.Ildl ..d KeareU Metrlp L7ah b.d bees ~.ben .{.oe the tao.rd was •0 saaM.wo M 1 ~x., r/.uM an. a.Myw a~ ..o F+.a 6+os 7vo ...c..  u. , ~.rir oora+.l yd ...t axperir.oe of 16w .e.{ot ..cs of .ckocs sd roedkios did awc.b b.d...a. Th. CourdPk rescuc% proW.... . 'iL. CancY Y t!M spoa.ari.g q..cy of a progr.n of «acar<L Iao que.t.b.s ot tob.coo ur ..A YeaNi It I. t!0 oytsro.th of .n orp.lsalo. ~ r forwr/ s.rfy M 19!• by nlrve.tvd.w of lob.oco na.u(icturva, lpo.a+ ~ t J, w•aelw..anes. Rc.eatdl wrpport Irm oen a.i.ly tbron& a pro6rae of ~. .~ ~ 6rr,.t.lo•aM wpple.e.ted by coancts fot tatarcA .ilb LMYixloo. .ed {.bor.wriez Ths Ca.eil doe..ot iudf opae+N u7 rese.rch f.dUty. j. .• T!. Sekati6e AQri.a7 Re.rd 1o Tlr C•otr.cY seas eesulady to o du.t. ..pq+lat(ons for „aw•it.ld ..d for oo.a.M )+rdyy aoa .okty os th. '•!..ls of .d.ai6c merit rd rdera.co. TDo Couotil .watd. nsbrt! Sno1. /o iod.t..deM akftists who .n ., ..wad ooe.pkte ai.otjfk re..da. 1n oo.dr.t!!.6 tladr Nudie.. Uraaees atoeis ;~ .n fsyorrihN for r.portrj or publlsbi.6 t,di fi.dier r t!r .cccpte0 .deMuk e,.ser - IDro"e .wdic.d ..d .deatuc jour..l. ,.d .ockue.. Throu6A Dooeatber 1973. nsearcb prooct. \.re beeo .ppro.ed f u 299 ~L.estlo.tory I. 2111 roodkal .eAooi., f,o.'itaM ..A eaarrJ lo.tkatben. Thw .-.r& totaled ww+ th.. f26.000.000. 71ThV repor~ iedrda a l.id .r.rnt.ry of T1. Car.dl'e praeet pro6r.+. .. .eM .. 8Ms of the cwr..t ..A pe..brr nr.anh proec1u wppowled by T1w CarcR. Also Mclud.d .r+ abstracts of I 16 nre.rdr p.pam .ck.o«lodtloti ,,Carefr .uffart that ww* publi.Md {..eMNfie jorr*d& drriy 1973. A total "d 1,170 .reb ppers l+.s e..e publli.d by proiKt nelv+ru. ~ m ~ H. H. Ra+.r (ldte..,n yd Pre.1d.ot 1973 REPORT 0/ TIIE COUNCIL FOR TOBACCO RE5EARCI$-U.S.A., Doe. Tl1E (Y1lINC11. F'11N T(1RMf.C(1 RI:SF:AR(71-II.S.A., Inc. 110 Fa.t •Syrh titrert, New Y.rV, N.Y. 10022

SCIENTIFIC ADVISORY BOARD to The Council tot Tobacco Research-U.S.A., Inc. as of December 31. 1973 SHELl7nN C. SOMMERS. M.D.. Chai.nsan [)irrctor of L.uM,rarorirs, Lcnoa Hill Hospital (7inical PrnJrssor of Pathology ('olunsbia Univcrsity Colk`e of Physkians and Surgeons New York, New York HOWARD B. ANDERVON r. Sc.D. Scirnri/ic Edit,>. (retired). The lournd of the National Cancrr Institute Bcthesda, Maryland RICt1ARD M BING, M D. Director o/ Cardiology and Intranrurd Medicine Ffuntington Memorial Nospital, Pasadena, California Pru/nsor of Medicine l)nivcrsity of Southern California School of Medicine Los An6eles, California WILLIAM ll. GARDNER, PH D. ScirntilSc Dirrcrur, The Council for Tobacco Research-U.S.A., Inc. E. K. Llunt Pro/rssor of Anatrwny (retired) Yale University Schrx>t of Medicine New Ilavcn, Connecticut RC)BF:R I 1. HUFBNER, M D. ('hia/, Viral Carcinosencsis Program National Cancer Institute Bethesda, Maryland I.EON O. JACOBSON. M.D. L)ran of the Division of Biological Sciences RrRrnstrin Pro/cssor of Biological Sciences lJnivcrsity of Chicago Chicago. Illinois AVFRII.L A. LIEBOW, M D. 1'ro/rssor and Chairman. Ikpartment of Pathology University of California School of Medicine San Dic6o, California III:NRY T. LYNChI, M D. 1'rn f r s trrr and Chairman Ikpartment of Preventive Medicine and Public llealth ('rerRhtun (Inivcrsity Sch«.) of Medicine Onraha, Nchra%ka I NANS MEIER. D.V.M., Dr. Mcd. Vet., M.R.S.II. Srnior Sraff Scientist 1 he Jackson Laboratory Bar Harbor, Maine IONN P. WYATT, M.D. Pro/cssor and Htad. Department of Pathology University of Manitoba Faculty of Medicine Winnipeg. Canada sclewrift S1a/f of 11.. Couaell WILLIAM U. GARDNER, PN.D. Scientific Dirccto. ROBERT C. IIOCKE7T, Pu.D. Research DGrrctor JOHN li. KREISIIER. PH.D. FREDERIC W. NORDSIEK, PH.D. Associate Rescarch Director Associate Research Director VINCENT F. LISANTI, D.M.D. Research Associate

Introduction Introduction CONTENTS Cunent Status of the Rescarch hopas ....... 5 7 Canctr St ud ies . . . . . . . . . . . . . . . . . 7 Chroek Respiratory Diseases . . . . . . . . . . . . . 9 Cardiovascular Diseases . . . . . . . . . . . . . . . 13 Pharmrnlnev and Psvchooharntscoiotv . . . . . . . . . 13 Epidemiolotty . . . . . . . . . . . . . . 14 Abstuacts o/ Reports . . . . . . . . . . . . . . . . 15 Studies Related to Cancer . . . . . . . . . . . . 15 Heart and Circulation . . . . . . . . . . . . . 28 The Respiratory System . . . . . . . . . . . . 34 Phumscdc+Ry utd PsychopharmacoloIlly . . . . . . . 51 lmmurxAM . . . . . . . . . . . . . . . . 61 E pide m iob6y . . . . . . . . . . . . . . . . 65 The Normative Aaing Study . . . . . . . . . . . 67 M iacell uKou. . . . . . . . . . . . . . . . 75 Active Projects . . . . . . . . . . . . . . . . . s0 Compkted Projects . . . . . . . . . . . . . . . . 66 Index of Senior Awhon . . Index of Principal Authors . . . 94 The interaction of the genetic or conslilulional factors and the envkn.- mental conditions to which an individual is exposed over the course of m.r.y years probably contribute to the expression and nature of the ying-associued diseases. Those diseases thot have been statistically associated with smoking and health are predominately age-associated diseases. In some instances, an Indl- vidual may have an /nherited tendency /or a disease, for example empAysema, and the associated .nd presumably conlribMrory inheriled dpha-l-anti/rypue defkiency. During the p.st few years much has been learned of viral eapression and cancer in laboratory animah and of chemicals which, when modifkd by entym.s that may be prexnt, can transform normal body cells into cancer cells, po.sibly through initiating viral espres.ion. Chemicals thau may thentselves be quite i.- active In Inducing cancer In laboratory animals may make srnall and ineffectlve amounts of known cancer-causing chemicals capable of transforming uorwr.l cells inlo cancer cells. It is now possible to monNor the environment of iebrad laboratory mice so that the viral, chemical, immuoological, and genetic qualiiea can be knowingly controlled and varied. The env'wonmenl of laboratory animals can be controlled more apeclYcary than can man's environment. Even when genetically eontrolkd and predisposed laboratory animals are used, exposure to known or presumed usar-Invdi.~ ehemicats mual eRtend over prolonged periods. Only morwrsygodic (ideMicall twins afford a genetic unlfonwNy is w comparable to that of inbred laboratory animals. Il they are disoordant 1. dr` terminabte environmenul esposurn, these twins could provide data that may lead to the determination of the relative Importance of genetic and eaviror• menld lacrors. Such studies art being undertakew in countries in whicj dr mortality from lung cancer reportedly differs more than fourfold. The transformation of cells in tissue eulture lus been studied i. Iwpes of obtaining a rapid and reliable method of dqeeting small amounls of ea.cer- causing chemicals in materials to be tested. The occasional "sponta.eaua' va,..- formation of cultured cells, the'resence of 1a1eM viral antigens and odser qraY- ties have not permitted etclusive use of cultured cells for ddection or qwdMa• tioe of cancer-causi.g .ubuawces. Up to this tinse, the resd/s dlirwr, ir Rencr.l, those of experiments with intact animals. The in rlrro techniques pwvide the methods of investigating mechanisms of cell transformation at a aubcetlular or molecular kvel and of search for inhibitors of translormation. Revcrslon of transformed to normal cells and selective inhibition of growth of uaa+lormed eells can also be deteqcd /n vitro. The discovery that the kvels of inhibilors of protcolylic enzymes ie rhe blood of persons highly susceptibblo emphysema are low, has reveald Y kaM one objective hereditary predictor quality for this pulmonary disease. Pursher- more, informarion on the sources of prote.ses, the mechanisms of rher aklion on body tissues and rhe characuri.rics of their Inhibitors continue to be .ow.er. The affe-associared atherosclerotic vascular diseasn, panicutarly those in- volving the crwonary artteies, continue to be /nvesuigaled from nceir sescrk and environmental rntcrrelatwnshrps An enzyme that makes cl.oksarod nsur• soluble, lecithin cholesterol acyl-transkrase, may provide informalion /oc slgnifi- eant advances in alherosckrosis. 5

Nicotine and its metabolites, under sonx eircumsunces, have Jircct or in- Jireci cllecr, upun nerve cell.. lhe development of raJwirnmunuas.ay methuJs for the Jeteclion of nico/ine and rneuholites is lacililahng work on rncratwhc studies and nuy rcveal speafie nerve eells upon which they may act or may have binding sires Significant advances in the psyehopharmacolngic.l mpecb of smokrng may he eapectcJ as mNhoJs develop to permit eapbratiwr of a dwect nicotine lnnJrng t cellular or subcellular kvel. Some of the renearches supported by The Council are at the cellul rr and suhccllular levels It is mtportant that advances continue to be made at such methodological anJ conceptual kvels /o assure continued progress in other aspecls. The support uf hasic research in Immunology or virology, for easmpk, may contribute signdicantly to an undenlsnding of smoking and hcalus The more than 100 papers reporting investigations supported by The ('ouncd that were puhlr.heJ during this pasl year reveal the breadth of the overall reuarch interest *u t usr l1. (3aat»rr-a. PN 1). Scienlilk 1)ircclor Current Status of the Research Program The research program of The Council for Tohaeoo Research-US.A., Ioe. espanded funher in several directions during 1973. with studies eelatiot so cancer still receiving the greatest emphasis. Studies of chronic pulmonary diseases and epidemiological investigalioen have also bee. eapanded. Among the latter, the study of identical twin populations as a nseans of separ.ti.j gewetic from envvonmenul factors in Ihe genesis of diseases has been ettendcd. Researches on cardiovascular d'nease" and on phurn.eolopr and psycho- pharmacology were continued at approaimately previous kvels. As herelofore, Ihc emphasis was mainly upon the etiology or pathogenesis of tl,tse aRing associated eonstitutional diseases with a special iotereu in the discovery of factors predictive of suseeptibHity or resistanee lo them. Since the published results of Council-sponsored studies are .bstr.cted elsewhere in this Report, our intention here is to give perspectives on tbe nature and eatent of the program as a whole. the distribution of emphasis arnon& its parts and on the purposes and plans of approach in such a way u to aid in fittrng individual research reporH into a conmest. It will be recognized that many studiea acknowkfting Council aid represent small methodological steps toward ultimate pals and Ihal some arc by-products of the main cflort. Canccr Studies During the year just ended. The Council concentrated major attentioa upoa an eatensive, systematic, multilactorial and step-wise study of carei.oge.csis in a number of contrasting mouse strains. The object of the ongoing study is to define and measure several genetically mediated biochemical characteristin that are thought to determine and predict the responses of these animals to eaternal "carcinogenic influence.," especially to chemical substances introduud into the lung with or without promoden or special conditioning. This under- taking is to discover and define conditions under which truly malignant, aggressively invasive and lald squamous carcinoma of the lung, with mctastaae., analogous to the human disease, can be obtained repetitivcly and predictably in such animals. l he importance ol this goal is gnphasired by eaperience showing the ua- certarnries in diagnosis of nsicroscopic, incipient lesions by purely rnorpholopcat criteria and in the use of such ksions as end points in eaperimental reu.rch. 1/ this aim is naineJ, in the contcat descrrbed, it should provide in/or- mation in b/ochcmKal, tenclK, viral or immunologic terms how the snimal. that develop this dIKaY diflcr Irom those that prove to be reswsunt or immuoc lhrs n the kind of rnlormarion Irom animal eapenmcnts JccrncJ rewst likely to provide promising clues /ot spplrcatuon to human studies 7

/nadrquocy ol MouscS4in Peintin6 llx ('ouncil's often repeatcd disutisfaction with the conventional mouse- skin painting with amoke condensales as .n indieator of "tobaeeo carcinoteni- eity" uems in good part from the well-known and very great differenus among •nimal Mrains and species in their responses to treatments of this kind with known end paenl chemical .gents. Without information about the biololical bases for such diRerences, any arumption that the resuNs of such mouse eaperin.ents can be estrapolated direcdy to other rpecies, especially man, is highly presuraptuous. A('ouneil-sponsored study completed during the year employed wrousaskin esposure so whoM fresh smoke in Its aerosol fonn in comparison with skin painting with .rnoke condensate from the same refaence cigarettes. Tl+e results appewed to show that Me condensates produced hothly utifactu.l re.ulu. If onwfirmed, this study noems likely further to reduoe eo.fldenoe in dse skin painting teehnlvw. Similar uncertainlics pert.in to the signiRcuce of nearly .U smokc in- halation studies heretofore curtied ou1 with animrda because of {nadce uata deinitiow of the animals and poorly controlled or unquantitated etpAUre condilio.s. For the current nMrhifacyor1 al prolect, the tnouse h.s been the spxks setected because it oAers the n.asimum potential i. (arms of definabk renelic .nd bioefremicAI susoeptibility or resMa.ce factors and thus may hop fuUr provide the shortnt disces.rbk route toward huma• relevance. 11 should be under.wod that the highly Inbred mou+e stralns used in the study rue euen ~ially laboratory ereations end never esisted as such in the undisturbed ata e of nalure. They must therefore be regarded as "bundlcs of traits;' not as rlirect models of man, ao that caperimental results will be rclevanl to man (or Hhcr armah) only inwfu as the same traits arc present and simrlar in import. "Fteder" Studies Carried Out The present project became poasibk only sfter a large number of pre- lirni.ary or preparatory bioio=ical projects had been carried out as "-fecdcn" to provide basic information essential to Its design. Previous Annual Rrports have describcd or listed many wrch investigations. The program b uitl passing through syslenulie, step-wise developmental phases including subcutaneous injectio.s and lung Implantations of know. "eareirwgens." When suf:icient informatloo about susceptibility and resistarwe to such carcinogens hat been aocum.tasoL {ong-term, chronic espowrm of contrasting "de6ned" animals to inhalation of whok, fresh tobacco smoke or its gas-vapor phase are con- te.rwpiated. llrcu will require the production of smoke under conlrotled con- ditions, from tobacco products of known composition and chuactcristics, delivered quantitatively and monitored by tissue dosage measurements A battery of tcsts and observations is being developed and refined for application to the treatcd animaM. Several snvrec contracls for carrying ou1 the necessary ehemical, physico- chcmical, analylK, mcchanKal, •nd calibration work conlrnue in operation at a high Icvrl to support the dcvoon of the eaperimcnts In such carrfrJly cwwrdkd tong term amokc inhalation eaperimenls, using hithly su.crprihlc mNc of lahu.atory dcs«gn and other strains with contrasting combinatiorn of putative susceptibility characteristics, either the appearance or non-appearance of squ.mous lung carcinoma (or odher canccr), should provide, nor immediate conclusions, but scienlifie information on a new plane of interpretability and potential relevance to man- Biochemical and 6enetic F.ctors in Cancer Among the biochemical or genetk factors rekvant to cancer wsupibilky or resislance in mice, three may be mentioned here• (1) the Renaically- medialed presence of aryl hydrocarbon hydrosylase (AHH) i.ducibility, wbich determines the ability of the anim.) to convert various inactive but polestLl cucinogenic substances into an active form capable of attacking the cellular nuclear rnaterial, (2) the genctically, determined esptenions of Indigenous "C-type viral genome," and (l) competence of the Immune system and iu ausceplibrbty to dcpression by "carcinojeatic" influences or agents. During the pau year, ('ouncu'-sponsored stud'w reported evidence of a genNie relationship between susceptibility of mice from several strains 1o )-methykholanthrene (MC:A)-induced arbcul.neous Iunwcs sd the ioducibJiy in these mice of aryl hydrocarbm hydrosylase. Inducibility was controlled by a single .utoe:ornal dominant gzoe and reflected in eve-to-ten timea greater MCA susceptibility in "indueibk" animal.. These repons stimulated other investigators to study AHH levels a.d inducibility in m.n, with tesuks su{Resting that {n man also, inducibdity in controlled by a single (incornpkuly) dominant gest. This,resuha in a diuribu- tion of inducibility in a random population into three groupinp - low, Wer- mediate, and high - essentially in accord with theoretical prediction. A preliminary study of AHH levels in human lung cancer p.tieeb cated tnu nearly all the victims of this disease in the aampk had either iMer- mediate or high kvcls of AHH inducibility. The Council has uodertakes to support eflorts at simplifying and etpediting the tea prcceduren for use 1n clinical studies to evaluate their potential In aresung cancer susoeptibiluy or resistance in human populations. Other cancer investignioas, collateral t;o the projeq describe,d, included: in vitro studies of malipust tr.nstormalion of iung epithelial cells and of possible induction of squamous metaplasia in ecUs already maligaaat; efforta to identify the agent or agents thAt produce .heep-luog .denomatosia and to transfer these to miee; Immunological studies of tumor•auoei.ued stipens in human cancers of ovary and hrnot; tesearch into pfasm.-membrane associated UNA; genetic studies of oncogenesis and vertical virus transmissiorr 1s the rabbit; mechanisms of suppression of cellular immunity by carcinogenic hydto- carbons; work on the role of cydie AMP in carciaoscncan; esplocation of melhods for enhancing the immune responses to neoplastic lissues; and evalu.- tion of chromosome changes as indicdbra of m.lignant transformation. Chronic Respiratory Discasts 'Ihe lung u a unique organ. It is a double structure with two very similar halves in practically constant motion, eap.nding and contracting in a uniaon so perfectly balanced that it would appear to defy the laws of physica. We 9 0

know that rhe balance Is possible only because of an internal coaunt by a matenai, the "sur/actant," that equalizes dd(crences in tension that would orherwise result hom dif/erent degrees of distension of the air sacs. In addition, the lung has an enormous internal surface of thin mcmbrane, equivalent in an adult man to abow hal/ the area of a tennis coun. The entire blood supply of the body must be transported a1 frequent intervals to one side of this membrane srr/acc, spread over Ns es;unt and then earried away, whde air is moved in and out from the opposite side for absorption of otyten and removal of carbon diuude and other substusces. Thus the lung is tremendously involved in transport of materialt tts renieal position in man, contrasting with the horizontal position in most other rw.as.b, alters the eflects of psvity and creates certain problems. Besides thene mechanical operations, the king and bronchial wrlaco are eaposed lo a great variety of backria, virsues, duas, Sases, smokes and Hhcr ealraneous muerrals that musw be kifked, reteowd, detosifkd, inactivned or otherwise disposed of. The lung has renurt.ble tapacitiea for thex functions. ( hromc diseases of the lung sueh r"bronchitif" and "emphysem" are disocden that tenerally develop BraduaMy .ad progress over a long pcriod, wtgcstint that they have their basis in a gradual lou of efficiency of otre or rssore of the mechanisms mentioned. Their association with the aiting process and a tendency to "run in (amilin" aho suqesu that some congenital de- ficrency in one or more of the functions described may prtdispose to rela:ively early development of these disorders. The pathogenesis of Ihese diseases, that is. the successive steps in breakdown of normal mechanhms which f nally bring them to recognition at the clinical kvd, has been very obscure until recently and still presents a great chdknte to science. They are not by any means new diseaurs but they have become more prevaknl as causes of illncss, incapacitation and dearh, s the acute fatal infectious diseases of Ihe past have been elrminated, allowing potential victims to reach a more advanced age. Neither bronchitis nor emphysema occurs naturally in animals in a form that could hinceno be regarded with assurance a duplicating the human disrKden and thus serve as a reliable medel foe e.petimentation.' Although "chronic bronchitn' and "emphysema" are separate condrtions, each occurring in several distinguishable types. they are often associated ~n the same individual, which complicates the probkms of etiotogy and pathogenesis. ( hronic bronchitis is necessardy deflned in clinical rather than in path- ological terms, chiefly because uncomplicated bronchitis is seldom .ccn by pathologists in the normal course of events. Tl+e disease, or syndronse, is characterized by recurrent acute infections of the lower respiratory lract, especially in winter, by mised fSora of v'raI and bacterial agenb, and is tcncrally accompanied by chronic cough and abundant production of mucus. Ir is thus unlike the acute infectious disea~[s caused by specific organisms acting in a speuRc manner nd running the familiar type of corrse to death or recovery. Biopsks have %hown over-growth and activity of mucus ucrcting cclls in bronchitis Neverthekss, the nature of the basic defect has not really been estaldi.hcd, whether eaceu mucus, a defect in the germicidal syucm, an auroimmune manifroutinn, or u.me other factor. Whcre thc diuau has become edahlished. inhakJ environmental irritants may well at1ravaie rhe symptoms While theie is an rmpresuon of long standing that familial pre- ditpusiuons may tu%t. the various clinical entities grouped under the term "bronchiris" have not so far been clearly enough distinguished to permit clear separation of genetic from environmental influenoea. Tobacco Smoke Inhalation Studie. As one approach to the bronchitis presMem, The Council h.s maintaieed long-term studies of the effects of many factan, including tobacco amok• inhalation, on the disposal of living bacteria Implanted into the lungs of mioe. When bacteria are inhated, a mobilization of white scavenger eeUs (mauo- phates) into the lung b stimulated. TBess white cells engulf awd datroy ma.y types of organisms and foreign particlo. Ttse Msdies with mice have show. that inhalation of tobacco .moAe produces a similar mobilization of macro- phages. Though some of these defense cells .re isactivated by eootact with the smote, the available numlas are so Bready increased that the overall capacity for bacterial destruetioa is nol diminished. Similar studies of macrop,'latcs recovered fran the lungs of human subjects after ,moking have sisows that they m.laais a high bacterkidat capacity despite some changes in appearance and in oraygea requirements. These observations do not support the hypothesis that smoking oonuibutes to development of bronchitis by depreasioe of the tn.aoplute defense apiw infection. Emphysema is more definobk th.m bronchitia in physido&al or path- ological terms. Basicaily, it is a:a isaeaae in the site of the air sacs (alveoli) beyond the terminal brondsiotes (a'r ducts) due lo los of elauicity or actud derruction of the alveolar waqfl. Obstruction of the terminal broochiotes may contribute to these processes. E.nphysema can either be difluse throughout tbe lung or located in "islands" distributed through woneal tissue. (t tends to be propessive, with increasing hyperinBation, airway obtructioo, and residual lung vdume, defective muinU of psn, and eventual bteathkssness and oaytet starvation. The tissue destruction, once it has occurred, cannot be repaired. Emphysema can be diagnosed .rutomically, with measurement of the type and eatent of deatruction, by postmortem e:amin.tion of lungs suitably inflated and fised. ('linical diagnosis has been more difRcult so that death certilkales in the past have made poor distioction between bronchitis and emphysema and also have provided rather generally unreliable classifkatioos of either disorder as to type. Greatly improved methods of measuring the mechanical functions of the lung and more sophisticated radiological techniques are now being employed increasingly. Ensyme. T/rot Destroy L,ung Tissue Because damage or destruction o( the elaslic structural substances of Ihe lung is a basic feature of emphyserna, it has long been surmised thal entymea capable of breaking down these aubsunces, p.rticularly elastin and eollaeen, might be involved T his concept hst been supported by esperirnenu in whkh the Injection of vuKws "loreign" proteolytic ensymn into animals w.. (ol• lowed by lung dcstruirion mrxe or ku resembling human emphysema At present a great deal of study is being concentrated upon the search fur enzymes capable of destroying lung tissue that might be present in the lung under real life condnions. Rre lung itself produces protcases, particularly II 10

during its embryonic uages, and the sature nd functions of these are being investigated urMkr a new Council grant. Moreover, the macrophages and small lymphocytn, which play a major role in defensc of the lung against infeaious organisms, produce and utilize proteolytic enzymes in the process. Proleascs, released by disintegration of these defensive white cells or leakage of enzymes from them, can themselves damage the lung. Normatly this is prr-ented by enzyme inhibitors such at the atpha-J-aetitrypsin that circulates in e.se blood. 1 he emerging picture is one of a delicate balance between protealylie and anti-proreolytic activitiea in the luep searay for its defense against infection but capable of damaging its slruettre IL t!!e balance is disturbed either by over-productioo of proreases or trder-}rodut:tioe of inhibitors. A number of the studies curteotty being spoasored lpy T)a Couacil, as wilt be eviJent from the listing of current projects, art twnarned with IueR prote.ses and their inhibitors as possibly contributory to empbrser>v. There has been considerable publicity orer the discovery that one tuch enzyme iehibitor, alpha-l-aNitryp.ia. Ir eoyesiuMy defieieet ie .an: persoas and that such persom m:y be eapeeL/y saaceptible to empbysema. iDough the situation has beee oomplieaud by the discovery of several diHerxnt forms of aotitrypaie with diAeressces r activiry, k ervertbelese appears to be ermly esubiished that a hereditary predispoahio, does eaist. Oee Couocil-aponswed project is studying the possible relations of the several genorypes, as determined by blood assay, to responses to ee.lro.nkatd stresses In a large human population. This study has also produced evidence of familial predisposuioos other than thoae suribueabk to antiuypsie defkkecy. Another hypothesis, not .eoeaarJy (e conflict with the proacase-aoti- proaease coocep, views lung tissue destructioe as due to changes in its aoti- teoicity that invite attack by the body's own "foreiRn Iissue° rcjectioe mechannm. Such changes conceivably could be produced by relatively slight chemical aturatrons of the lung structural substances by inhaled pollutants. ('ouncil-sponsornf srudies based on this hypothesis continue to produce ws- gestive results. As implied by our comnenu on the function of lung surfactant, a de- /kiency or maldntribMNion of this agent would be cspected to produce significant matfundions of the lung. A Council-supported invesliptoc sucoeeded in adapting certain physiological lung functioa measuremesu (oe application to saull rurimals, including living mice. This promisn to be useful in eombirutioe with postmortem lung ex- amin.tiorss for studying age-related changes (e mice of different suains, in the search for a belter emphysema nwdel. Sucb function meawrements may also become a useful tool in determining whether eflecb of various inhaled agents, including tobacco smoke, are temporary aed reversible or persist nnd prosresa to the stage of posrmortem observation. 6esides the numerous funclioes meNioeed above, the lung also has metabolic activnies such as nmovHeg frore the blood atreans a number of hormones that ue active In the mediation of inflammatioo, io the regulation of blood pressure and in mobJis.tion of white blood cells. -two Council- sponsnred studies arc espkxing these subjecu. Seventeen projects In the field of chronic pulmonary disease s.ere active in 1971, Two of these terminated during the year and /our new unes were aplnoved for acuvaion early In 1971. Cardiovascular Diseases Notable progress was made during the year, under Council spoesotship and independently by other ievestipton. I, the deve{opmeet of 4tususo- prweise chemical assays for nicotine and its major tnetabotites. These show of providing a rapid, precise and highly specific method for way of ekods and relsted compounds in body Auids, especially blood plasma. Such ruethods, when perfected, should make possible the studies of nicotine pharmacodyeraio that have long been needed for solution of rtuny importaet prcbkms. Amons these are the kinetics of nicotine absorption from the various tobacco products under normal conditions of human use (ciprettes, pipes, ciRars, souA, chewing tobacco), the peak kvels to which nicotine rise, in blood, rates of inetabotism and eacretion, etc. The effects on these kinetics of genetic diflerewees, of habituation, of taking oommon household eredicinn, of temperature, esercLe, and many other factors should then become teeasurable. Application of such measurements to human populations could add axw dimensions to Ihe ieterpretation of epidemiological studies of cardiovascular disease in rclaUoo to various forms of tobacco tase by showing whether or sW nicotine can be ievolYed. Conferences with scientists from a number of countries aro furlb.r u- pbring the feasibility of international twie studies to test more rapidly .ed decisively the implications of Swedish researches on ideetical twins whfa dis- cordant smoking habits. These suueat that genetic factors are predosie.at over environmental ones in the genesis of evdiovascular diseases. Assistance to the Swedish study continued meanwhile. Aside from epidem3ofogica) studies bearing apoo cardiovascular diaeus, thirteen experimental or clinical studies were supported durios the year. Taer proceeded along the lioes described in the 1972 Repon, with a yield of wasy publications that arn abstracted ebewhere. Two of these studies Isaw now terminated. A new project was approved for study of possible effects of tobacco aawta inhalation and of nicotine upon development of coUaterd oorooary blood low following acute or chronic occlusioo of a oororsary artery io dop. Pharmacology and Psylchopharmacology This phase of The Council's program continued along the liees deacribtrd in last yeu's Report. Current emphasis is being placed upon aMeruioaa ie nicotine pharmacology that result from chronic e.posures through habilu.tio., adaptation or enzyme induction. (Aher pharmacological probkms await further development of the new analytical techniques to provide procise inlorsealion on the cuncentrationr of nicotine and of Its various rnetabutotca present at particular trnses in specific tissun and at specific sites. Studics of nicotine effectt on animal behavior are presently receiving little emphasis, not because there is any dearth of academically interesting problems that could be appeoached by eanring techniQues, but ratl.er because the new analytical mcrhods rnentroned as being in reasonable prospecr shoutd inacase the sophisticNion of any future studies in this arcam to addttion, there ar• th. 12 13

imporlanl probkms of whether and to what eslent the implications of rat- learning and bchavinral studies can be transferred to man and then of cvalwding their practical significance in human life. One new animal bchavior study of unique character is, however. IrcinR inaugurated currently. The investigator has ahown that a rat behavior IerroeJ under the influence of a particular drug X can be made contingent rpon presence of the drug state. Such behavioral patterns can then be used as tests to determine whether the rals wbjectivefy perceive other drugs as X, as nca-X, or as reinforcers or antaRonists of X. The elotts of nicoline and sonse of its analogs and metahotites will be studied in this conleat. A human behavioral study, reuelly begrt, will eaplore the eflccts of smoking deprivation on group problem aolvi.j processes in terms of r.peed and accuracy. Epidemioloff Including conferences on potenliah for Internalional twin uudies, aia eprdemrolugrcal investigations were active durins 1973. Tl+ese on-going projeets, described rn previous Reports, rve producing a large number of signiflcant and rekvant findings on the constitutional and behavioral diflerenon at many drflercnt levels among persons who have plaeed themselves by sclf-sekclion into the sevcral categorres of smoking behavior. As the numerous reports from these studies tcach publication they ue abstracted in the Annual Reports. Sevcral, at well as many by prcxluct papen, are included in this issue. Ruer, at Ilrr .e , t• 1'u 1) kcscarch Ihrrctm A l,stracts of Reports FnlNrwins are abstracts, approved by the authrxs, of reports on new rc- se.u:h aclnowlcJ6rng support /rom lhe ('uuncd that have appeared in scien- tific juurnils since publication of the 1972 Report. The name of the recpient is in itahcs. '1 he ah.tracts are grouped under these headings: 1. Studies Related to ('ancer, (1 Hear1 and Circulation, 111. The Respiratory Syslem, IV. Phartna- colo6Y and PsychopharmacoloRy, V. ImmunoloRy, VI. EpidemioloRy, VII. The Norrrmative Aging SiuJy, and VI11. Miscellaneous. 1. Studle. Related to (:nncer Mf'TAH()11(' ('()NVf?RSION OF BFN7()f.IPYRFNf: BY SYRIAN IIAMSIFR I IVfR MI('ROSOMFS AND BINDING OF MEIAHOIIIFS TO D!?OXYRIBONl1('I.EIC ACII) Binding studies reported here show that epoxides, phenols, and other metah.Ailes uf benrolslpyrene IHP/ formed in hamster liver microuxne systems may not just be end products, but may be further metaMibreJ to bind to 1)NA. Speciflcally, nalysis of inelaholites of generally Iritiated hent<s/s/pyrcne /'ll-HPI produced by a Syrian hamsler liver mierosonsal systens has revealed the presence of a number of dihydrodrhydro.y derivatives of BP including 4,Sdihydrosy-l,3-dihydrobento/a)pyrene. (klection of this melaholile is taken as evidence that the K-reRion I,S-double bond is acted upon by a micrrnonul hydroaytase to form the •,S-epotide of HP which is subsequently converted via eposide hydrase to the dihyJrodiol. Incubation of several 'll-BP merabolAes with 1)NA alone gave littk evidence for spontaneous covalent binding Ilow- erer, when hamster liver microsomes were present, a nsetaholite recently identified as 7,S-Jihydroay-7,SrhhydrobenroLlPyrene was found to covakntly bind to 1)NA to a tenfold greater eslenl than BP itself, suggesting that this cumpuunJ may be an irttermediale in the pathway leading to binding of BP to DNA in rivo. Borgen, A., Darvey, H., Caslatnoli, N., (-ror4er, T. T., Rasmussen, R E. and Wang, I. Y. Journal o/ McQklnaf Chrrn/irry 16(S):502-506, 1973. OOther .eplw.rf: U. S Public Hcalth Service, and the Cancer Research F'ursds of the llnivcrsiny of ('alifornia. Frorn Ihe Cancer Rescarch Institute and Ikparunent of Ptunruccurrcal (bem- islry, Ilnivcruly of l'aldornia, San Prancrsco. INI)II( 71ON 01: ARYL HYI)R(X'ARBON FIYI)R(/XYI ASF IN III/MAN SKIN 1 he +ryt hy.hrx artKrn hy,hn.yl.1.c whrch hydrosylares thcnrAwl 2lpyrcuc ( HP ) has been shown to be ntarkeJly rnJuccJ in aninuls e4pu.cd to IN)lycychc 14 15 .