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Council for Tobacco Research

Report of the Council for Tobacco Research-U.S.A., Inc. [St]

Date: 1972
Length: 54 pages
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I;i(TItA COPY REPORT Of THE COUNCIL FOR TOBACCO RESEARCH - U.S.A., Inc. 1972
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i 1972 REPORT ~ . Cr F-- ot U TIIE COUNCIL FOR TOBACCO RESF.ARCII-U.S.A., Inc. TIIE COUNCIL FOR TOBACCO RESEARCII-U.S.A., Ioe. 110 E.at 59tL Street, New York, N.Y. 10022
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SCIEN'1'Ih'll: Al)VItiORY 13l/A1i11 to The Council for Tobacco Rc.rarch-U.S.A.. Inc. t:~.. . M C.G as of Dcccmbcr 31, 1972 i . i ~ SHELDON C. SOMMERS. M.D., Chainnun Rescarc•h 1)irectur, Thc Council for 7'ubacco Rcscarch-U.S.A., Iuc. Directur of l.ahoraturics. Lcnox I fill I luspital KIiNNIiI ll MfiRItlLI. 1.YNC1i, M.D.,Sc.D., LL.D. ('haiu•rllur unc! l'rcplcssur lirncrinrs of 1'utholugy Medical CuIIcgC of South Carolina. Chartcslon, South Carolina ~ Clinical Pro f rssor of Pathology Columbia Univcrsity Collcgc of Physicians and Surgcons IIANS MI:IIiR, D.V.M., Du. Mt:u. Vt:r., M.R.S.II. L1..' New York, New York •Srniur .StafJ Sc irruitir ~ HOWARD B. ANDERVONT, Sc.D. Scientific Editor (retired), The Journal of the National Cancer Institutc Bcthcsda, Maryland RICHARD M. BING, M.D. Dir t f C di l d I l M di i The Jackson L:,boratory, [3ar I larbur, Maine JO11N P. WYA"t"I', M.D. Pru/es.rur und JJrad• Dupartmcnt of Pathology University of Manitoba Faculty of Medicine Winnipcg, Canada U or o ar ec o ugy an ntramura e c ne Huntington Mcmorial Hospital, Pasadcna, California Professor of Medicine Univcrsity of Southern California School of Medicine Los Angeles, California McKEEN CATTELL, Prr.D., M.D. Pro/e.rsor Emeritus o/ Pharmacology Cornell University Mcdical College, New York, New York WILLIAM F. RIL•NIIOFF, Jtc., M.D. llurud Meinl,rr Ernc•ritus 1'n,/c.%sor linu•ririrs of Surgery Johns Hopkins Uniscrsity Schcwl of Mcdicinc, Baltimorc, Maryland WILLIAM U. GARDNER, Prr.D. E. K. Jlunt Professor of Anatomy Yale Univcrsity School of Mcdicinc, New Navcn, Connecticut ROBERT J. iiUEBNER, M.D. Chief, Viral Carcinogenesis Branch National Cancer Institute Bcthcsda, Maryland LEON O. JACOBSON, M.D. Dean o/ the Division of Biological Sciences Regenstein Professor of Biological Sciences University of Chicago, lllinois CLAYTON G. LOOSLI, Pa.D., M.D. Hastings Professor o/ Medicine and PathuluAy University of Southern California School of Mcdicinc Los Angclcs, California ticirntific titrff of The Couucil ROUI:RT C. I IOCKGTT, Ptt.D. A cYir{g Sric•n t+/ic l)irectur J. MORRISON I1RAl)Y, M.D. JOHN N. KREISHER. Pu.D. Assucrute Sc irnri/u 1)irectar A.ssuciate Scientific Directur FREI)f:RIC W. NORDSIEK. Pu.D. VINCENT F. LISAN"fl, D.M.D. A.Vwriutr Scicuti/ir 1)irecYur Scicnrific As.uKVUt.•
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CONTENTS Introduction . . . . . . . . . . . . . . . . 5 Current Status of thc Research Program . . . . . . . • . . 6 Canccr Studies . . . . . . . . . . . . . . . • • • 7 Cardiovascular Discascs . . . . . . . . . . . . • . . 8 Chronic Respiratory Diseases . . . . . . . . . . . . . 10 Othcr Studics . . . . . . . . . . . . . . . . . . 1 l Abstracts of Rcports . . . . . . . . . . . . . . . . 13 Behavioral Effects of Nicotine . . . . . . . . . . . 13 Studics Rclatcd to Carcinogcncsis . . . . . . . . . 15 Cardiovascular System . . . . . . . . . . . . . 33 Rcspiratory System . . . . . . . . . . . . . . 52 Pharmacology and Psychopharmacology . . . . . . . 66 Mctabolic Studics . . . . . . . . . . . . . . 70 Chemistry and Biochemistry . . . . . . . . . . . 71 Epidcmiology . . . . . . . . . . . . . . . . 78 Immunology . . . . . . . . . . . . . . . . 81 Gcnctics . . . . . ... . . . . . . . . . . 86 Activc Projccu . . . . . . . . . . . . . . . . . 89 Complcacd Projccu . . . . . . . . . . . . . . . . 96 Author Indcx . . . . . . . . . . . . . . . . . . 104 Introduction The Council (or Tobacco Rcscarch-U.S.A., Inc. is the sponsoring agency of a program of rescarch into questions of tobacco usc and hcalth. It is the outgrowth of an organixation formed early in 1954 by representatives of tobacco rnanu(acturcrs, growcrs and warchouscmcn. Research support has been mainly through a program of grants•in•aid suppkmcntcd by contracts for research with insututions and laboratories. The Council does not itself operate any research fatility. Thc Scientific Advisory Doard mects reyularly to evaluate applications (or grants•in•aid and contracts for rescarch, using scientific mcrit and relcvancc as thc sole crilcria. Rescarch grams arc made by The Council to indepcndcnt scientists who are assurcd complctc scicntific freedom in conducting their studics. Gnntces alone arc responsiblc for «porting or publishing their findings in the acceptcd scicntific manncr-through medical and scicntific journals and societics. Through Dcccmbcr 1972, rescarch projects have been approved lor 283 invcstigators in 199 hospitals. universities and rescarch institutions. These awards totalcd more than $23.000.000. This report includes a brief summary of the present research program of The Council as well as lists of the current and previous research projects sup- ported by Thc Council. Also included arc abstracts of 127 research papcrs, acknowlcdging Council support, that have appeared in scientific journals during thc period covered by this Report. A total of 1,052 such papers has been pub- lishcd by project recipients. If. Ff. Raaw Chairman and President 5
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Current Status of the Research Program The rescarch program of The Council for Tobacco Kcscarch-U.S.A., Inc. has progresxd further during the last Ig months with a cuntinued major emphasis upon canccr, chronic pulmonary disorden, cardiovascular discascs, and thc "psychopharmacology" of smokiug, in approximately that order. 'Ihc actual number of formally separate projccta is at present somewhat Icss than the historical maximum, reflecting a dcgree of aggrcgation into certain ccntcrs for the advantages of interaction among various different skills and methods. Seventccn of The Council's current projects involve thc exposure of animals to the inhalation of tobacco amokc. Sevcral others have been under- taken with the obiect of including smoke exposurc as soon as the neccssary preliminary biological work has been suftkiently developed. In all these inhalation studies the aim is to so control conditions as to insure expoaure to 'Yresh, whok, normal" tobacco smoke or its gas-vapor phase in monitored doscs, in dcfirud animals of several strains and specics, usually pathogen frec and virus profiled, for time pcriods dctcrmincd by the aims of the experiment. In scvcral cascs this is a long-term or normal lifctime exposure. Cigarettes or other tobacco products of defined characteristics are employed. Concentration upon smoke inhalation experiments reflects the conviction that realistic studies must be based upon and evaluated by exposures simulating as nearly as possible those expericnced by human smokers. Snsoke Exposure Devices Development of mechanical dcvioos for accomplishing such exposures with animals has been an exacting, timeconsuming task, even after establishment of criteria for satisfactory conditiotsa. Though The Council has been sponsoring smoke inhalation research for 17 yurs, the recognition of these criteria has developed only with experience. The present proliferation of smoke inhalation studies in many quarters empharius the aeed for recognized standards to which all such studies must eonform, if a great confusion of contradictory find- ings in the scicntiflc literature is to be avoided. Two devices that appear to meet maoy or most of the necessary critcria reasonably well are now at hand. One is a relatively inexpensive and simple machine which should, hopefully, be adequate (or use with intclligent care where relatively modest numbers of small animals arc to be employed by in- vestigatun who arc skilled as biologiua but not ncctuarily as mtxhanics. 'Ihc otbcr dcvice is more costly but should conserve manpower when large numbers of small animals arc to be exposed or when the use of large anirnals requires a greater rmoke generating eapacity. A number of the small machinu have now been distributed to Council- sponsored investigators aad arc in use. Consequently, a considerable increase in the flow of useuch raults from direct smoke inhalation studics is to be expeetod. - Despite these dcvclopmeat; there are still many t:aks to be carried out to .lcscrdx mure fully and to mcasurc more accurately the performance charac- tcnsucs of these devices, especially as apphcd to animals of different size and dilfcrcnt rc.pirawry mcchamcs and behavior. Especially important is dovelop- mcnt ol txtter mcthods for measuring the actual amounts of smoke particulates dcpusitcd in the lungs of these several species and strains of animals under actual upcrating cunditions. Such measurements arc necessary for reliable dctcr- mination of dusc-responsc relationships. Means for carrying out thcu studiu arc being sought as a support for the existing and growing program of inhalation studies sponsored by The Council and, increasingly, by other agcncics. Esscntially the same smoke exposure devices can be used in studies targctcd toward cancer problcros, pulmonary dircase problems, cardiovascular prublcros, metalwhc problcros, pharmacological problcros, and others. In many cascs, parallel cxlxrimcnts arc being conducted or planned to compuc any effects of whole snwkc inhalation with those of the gu-vapor phase free from particulatcs. In all smukc inhalation experiments involving animals, lhe factor of stress is kCing considcrcd txcatrsc, unlike man, most animals inhale smoke involun- tarily while stationcd in dcvices that impose res(raint, noise, handling, and other disturbing influcnces. This requires the usc of "machine controls" exposed to all conditions except thc actual presentation of smoke, (or comparison with unhandled cage controls as well as with exposed animals. Habituation presum- ably reduces stress and is a consideration in experimental design. Biochemical monitors of stress arc also being explored. This problem has generated two major projects in its own right. Pa[hogeneaia Studies ~• The Council program includes a selection of promising leads into an understanding of factors that may be significant in the pathogenesis of the several groups of aging-associatcd constitutional diseases which are currently major causes of human morbidity and mortaliry, and also those reputed to be linked statistically with tobacco use. Topics sclectcd for study are those deemed most likely to reveal important steps in the pathogcncsis of these diseases that may be tested in human cpi- demiological studies or developed into new and more meaningful bioauay tcsts, either in smoke inhalation studies with whole animals or in organ or tissue culture systems of several kinds. By pathogencsis we mean the progroa- sivc succession of changes from normal, whether hutological, biochemical, immunological, structural, or other, that lead to or arc concomitant with disease conditions. Cancer Sludies Thus, in the cancer ficld, there arc presently six Council-supported studies aimcd at a bctur undcrslanding of how genetit facton control susccptibility or resistance, and how the so-called C-type viral genome may operate in eon- trtbuting to oncogcnesis. Such knowledge is not only necessary (or seleetirrg appropriate animal strains for smoke inhalation work but also tor defining indicators of IusccptrbiGty that can be applied to maa 6 7
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"I wo other cancer studics that do not prcxntly involvc smokc exposure arc in the ficld of imntunulogy and are aimed at learning huw the bodily mcchanisms that bring about the rcjcctron of grafted tissues and organs as "foreign" ntatcrial also operate in rejecting ncwly-furmed cancer cells. Thc work includes thc objcctivc of Icarning why this rejection system docs not always succced in destroying cancer cells and whether it can be stimulated without undesirable side effects. !t has been shown that this part of the immune system is depressed by several of the "carcinogenic" hydrocarbons and, if uritabk new and scnsitive indicators of this deprcuivc cffcct can be dcvcloped, sukfies arc contemplated for determining whether chronic tobacco smoke in- halation produccs any such depression. (An cxtensive contplcted study of immunosuppression by another method involving skin transplantation failed to show any effect by tobacco smoke condensates.) One relatively frequent type of human lung cancer is squamous cell car- cinorna, which is seldom found in animals. t3ecause squamous cells +re not rwrmaUy present in the lung. thcrc must at sonic time bc a nlctaplastlc con- version of other cell types into squamous cells. 11 is not known whcthcr this convcrsion occurs before or after the transformation of normal cells rnto cancer cells. A new project has been designed to determine whether exposure of living human cancer cc111 of other types to tobacco smoke will evoke squamous metaplasia. Animal tiloclela for Squanrou. l:elt Lung Cancer In medical science, discovery of methods for producing a good duplicate of a human disease in an expcrimental animal has oftcn provided a basis fur rapid progress in describing the pathogenesis of the disease and in finding methods of prcvcntion, delay or curc. It has been more difficult to develop such models for the constitutional diseases than for the infectious and deficiency diseasea because of the urong genetic aspects of susccptibility in the first. Several methods, howevcr, arrived at by luck or accidcnt, have been reported for producing squamous cell carcinoma of thc lung in animals. Two Council projects include modification of such animal models in the hope of producing the disease predictably and reliably and at a level adequate to permit systematic studies of the influcnces of agc, aex, strain, hormone levels, nutri- tional state, cfficicncy of the immune system, infectioru, and other factors on the incidcncc of the disease. Such information should provide guidelines for thc dcsign of better oricnted human studies. Cardiovascular Diseases As with canccr, cardiovascular discaaos arc of complex etiology that in- cludes many contributory facton, among which hereditary predispositions may play an irrsporunt role. The predominant influence of genetic factors has been further emphasized in recent yean by studies of identical twins with discordant smoking habits. Though The Council did not participatc in the sponsorship of tho uudia carriod out on a Swedish twin registry or the original extension of these studies to a twin collectwa in the United Statw, it is supporting czplora- uum ~wu thc Ixn.ihiluics of cnlarl;ing the samplcs through intcrn.Iiun:d co- ulkr:,wrn. the gu.J t. tu uhtain murc extensive and conclusive d:ua without w.utlnb Iur thc .low accumulauon uf murbidtty and mortality figures from the urigwrl s.,ntples. Mcanwhilc. the muluvariatc analysis of large existing human data collcc- unns „ producing new pcrspcctivcs concerning the reported statistical associa- liurn between smoking and the incidence of these diseases. The position taken by thc ('uromiftcc that reported to the Surgeon General of the U.S. Public Ilcalth ti.rvicr in 1964 was: "M+Ic cigarette smokers havc a higher death rate front artcry di%casc than nonsmoking males, but it is not clear that the associ- atiun hiu cau1:J significance." '1 his statement is supported by increasing evidence that self-sclcction of populatiuns into thosc who "like, need or wish to smoke" and those who do not, produccs a non•rarulum division into groups that arc not comparable with res{xct to morbidity or lifc cxpccuncy. Hcnce, attribution of experiential diffcrcnccs tu the samc lactor (smukrng) that served as the basis for separation is ntocssarily inconclusive. Almost half thc current Council program in the cardiovascular fichi, apart from the cpidcmiologica( work already mcntioncd, is conccntratcd upon the study of atherosclerosis or arteriosclerosis. Does smoking have any effect upon the initiation or progression of these conditions that lead to incapacitation or death? One current project includes smoke inhalation. Most of the studics involve or include research on the effects ol nicotine. In several investigations, the initial nicotine research is to be followed by whole smokc inhalation. Induced hypcrtension is also being studied as an additional factor in the total design. 7 hrcc of the athcroscl,crosis studics involve whole animal subjccts, two arc in vitro expcrimcnts using both animal and human tissues for species com- parisons, and one is a biochemical study of reactions in human blood. One of the animal studics, using chiefly primatcs, is concentrated especially upon the long-term effects of chronic stress in producing persistent hypertension with consequential cardiovascular changes, when other influencts are wcll controlled. Investigation of potential nicotine effects in this context is included. Carbon Monoxide Studies Another feature of the program is the inclusion of carbon monoxide studies. Chronic inhalation of this gas by primates is being undertaken to test a hypothesis that such exposure can hasten atherosclcroais. One study, at the cellular level in viuo, is attcmpting to show whether the processing of fatly materials (akin to those that coat Ihc artcrial walls in atherosclerosis) is affccted by nicotine or cartwn monoxide either separately or in combination. All thc major human epidemiologieal studies have shown that cigar and pipc snsukcrs differ very little in their cardiovascular diaeaae experience from nunsmokcrs dcspitc the fact that the smokei of thae tobaecoa have a relatively high nkotine content in a relatively alkaline envirooment conducive to rapid absorption in the oral cavity even without inhalation. Hcrsce, The Council has becn stressing for many years the need for precise quantitative and kinetic measurements of the relative nicarne absorption during the normal smoking of cigarettes, cigars and pipes. Analytical methods adequate for the purpose have been lacking. A Council project is endeavoring to develop an immunochemieal ~" ~' i S ..JU 8 9
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assay mcth.xl for this purpose. It is gratifying that the govcrnmcnt agcnucs have now recugnizcJ this need and havc solicited pruposals lor tht Jcvelop- nscnt ul such methods. 1f the nicotine rcccivcd from thcsc three forms of smoking /and from chewing tobacco) should be closcly simdar, the eptdcmio- logical Jiticrences in disease incidence among the users of these several products could not be attributed to nicotinc. Chronic Respiratory Diseases Historii:ally, sevcral of The Council's studies of short- or long-term tobacco smoke inhalation by animals and man have included an cfTorr to Jcscrtbc any observabk changes in pulmonary mechanics or altcrations in the cells or tissues. The intcrpretation of such obscrvations in rclation tu IwlmunarY dixascs has hcrctoforc been handicapped by the general lack of krwwlcdge conccrning the stagcs in pathugcncsis of these diseascs. /t has, thcrcforc, been considcrcJ ncccs- sary not only to sponsor studies aimcd u a better cGnieal distinction bctween thc scvcral human chronic pulrssssrrary disordcrs but also tu seek and fulluw clucs to thc prcdispositions and altgrationa that dctcrminc the course of thcir pathogcnesis. Apan from lung cancer, "chronic bronchitis" and "emphyscma" arc thc pulmonary diseases of special interest to Tlse Council. Neuher is really a single disease and, therefore, subdivisions into distinguishable clinical entities occur as research progresscs. The Council has contributed to the devclopment of clinically distinguishable subgroups through subsidy of humao-patient studies in depth. fioth these discases or groupa of discascs arc vcry incompletely under- stood with respect to etiology, which may well diflcr in thc different subgroups. Epidemiological studies in the past have been confuscd by poor chnical dis- criminalion among scvcral entitics. Nereditary Predisposition to Emphysema Neither bronchitis nor emphysemaa occurs commonly in animals in a form known to be clearly analogow to that encoumercd in man, so there is still uncertainty with rapect to the srutability of any proposed animal model for studies directly relevant to the human problem. However, several promising approaches have dcvcloped. One is the discovery that certain human individuals have a hereditary predisposition to emphysema due to a congenital deficiency of a crreulating antitrypsin which functions as a protector of the lung against protcolytic attack by exogcnous or ersdogenous agents. The implications of this ftnding arc being pursued by study of a large human population. Extcnsivc health anJ employment histories arc available for nscmbers of this population who arc also accessible for blood assays of anti-protcascs. In the light of very recently developed information on genetics of the dcftcicncy, this study will attcmpt to relate the aevcral yenotypca to dilfercrsccs in resporsae to cnviron- rssenW inlfuences. Animal worK, irscludiraj arnohe inhalation study, is capitalizing on this discovcry by obacrvatiosr to deturnioc wb,ctbcr changca in anti-protcaao icvcla 10 may correlate wuh dcvclopmcnt of cxpcrimcntal emphysema or may be afiecteJ by ugarctre smokc inhalation. '1 herc is alsu recent evidence that emphysema may be an autoimmunc dlKasc, caused or aggravaled by chemical changes in the lung that cause lung tissuc lu be rccogmccJ by lhc immune system as foreign material and, therefore, subjcctcJ to dcstructivc attack. This concept also provides possibk approaches to dctccuon of those indiviJuals who arc especially susceptible to emphysema and to ways of testing whether cxposurc to smoke makes any contribution to changcs in the amigcnicity of lung tissucs. While the ctiolugy of "chronic bronchitis" in humans is still quite obscure, il is considucJ very probable that an impaircd capacity to clear bacteria from thc lower rcgiuns of the lung is an intportant aspect of the disease. The Council long agu undut.wk uqplxut of studics lu nseasurc the rsta of disappearance of viablc bactcna /rom thc lungs of micc after quantitated implantation, as affected by ;) numbcr of facturs including whole smoke anJ ps-vapor phase inhalation. 'I hcsc studics wuc fullowcd by others concerned more particularly with the pulrnonary macruphagcs as the probablc major cffcctors of the clearance proccss. Uoth in vivo and in vitro smoke exposures have bcen madc, and are continuing, to assess thc total functional modification of this protective system whilc avoiding the artifactual situations casily creatcd by exclusively in vitro experiments. Nescarch findings about the protective function of the pulmonary macro- phagcs have led to other Council-supportcd research on the operation of these and rclatcd defensive systems. At the end of November 1972, an informal rounduhlc conference was arranged by staff at the request of the Scientific Advisory Board to The Council. Mcmbers of the Board and staff, investigators conducting studies on pulmonary defense mechanisms, and several guest scien- tists met to evaluate progress, exchange information and generate new ideas. Odier Studies Psychopharmacology The Council is currently supporting hve studies in the field of psycho- pharmacology that arc directed toward lurther elucidating thc paradoxical arousal and tranquilizing effects of nicotine and its facilitation of the learning process in animals. Chronic effects of nicotine exposure have now been recognizcd in the sense that a shift has been reported to occur with time in the rclativc sensitivity of particular areas of the brain to nicotine as well as to othu substances. It was predicted that this shift in the brain area of major stimulation should have behavioral conscqucnccs in the prornotion of "purposcful;' as op- poscd to random, activity in habituated rats «uivinj nicotine. This prediction appcared to be upheld in tcst experiments. Because human smokcrs ordinarily receive nicotine chronically, it can be rcasoncd that pharmacological expcrimcnts with naive animals may produce results that are not wholly relevant to the usual human situation. Hence, a ncw emphasis has devcloped concerning habituation effcets on the psychopharma- cological responses to nicotinc. 11
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Et.idemiological Studiu Four cpidemiolugical studies are in progress which involve the analysis of /arge bixfics of data already collected by other agencies. Onc is a continuing invcstigation into which it is possible to introduce new tests upon the samc subjects about whom a great deal is already known. Emphasis is upon the basic ditTcrcnces, at many lcvels, bctween those persons who have chosen to smokc cigarettes and continue smoking, those who chose to smoke but lalcr discon- tinucd, and those who never adopted the practice. Comparisons arc also being made of the constitutional characteristics of cigar, pipe and cigarette smokers. One study provides comparisons between men and wonun, whnes, blacks, Orientals, and Spanish Americans. Basic differences that distinguish thesc several groups in relation to smoking practices are relcvant to the qucstion whethcr sclf-sclcction invalidates or qualities the causal interpretations that have bccn placed on statistical correlations between smoking and diseasc. It smukcrs and nonsmokers arc diffcrent kinds of peoplc, there may be basic constituuon.l ditlcrences in disease susceptibility unrelated to smoking practiccs. Thc datn collccted also contain material for many other multifactorial studies. Metabolic Studies Two studies on metabolism are looking into relations between smoking and protein metabolism and into the possible influence of nicotine and smoking on the metabolism of various drugs, as well as the possible effects of drugs and mcdicinals on nicotinc mctaboliarn in the body. The current program represents a conccrted c(fort by the Scientific Ad- visory Board and the scientifie staff of The Council to achieve a new and higher degree ol integration among its elcmcnts and a sharper focus upon key prob- lcros in the area of tobacco and health research. Robert C. HockcN, Pu.D. Acting Scientific Director Abstracts of Reports Fulhowing arc aharactc, approved by the authors, of repons on new re- scarch acknowledging support from The Council that have appeared in scientific journal% since publication of the Report for the year ending June 30, 1971. The namc of the rcupient is in italics. 'f hc ahstr,crs arc grouped under these headings: 1. Behavioral Effects of Nicutunc. It. Studies Related to Carcinogcnesis, 111. Cardiovascular System, IV. Kc.piratury Sy.tcm, V. Pharmacology and Psychopharmacology, VI. Mctabolic StuJics, VII. Chemistry and Uiuchemistry, VI11. Epidemiology, 1X. lmmun- ulugy, and X. Ocncucs. 1. IleJiuvivral E,Q'ects of Nicotine S7'(JI)Ila ON TIiE MECIIANISM OF AVOIDANCE FACILITATION 11Y NICOTINL Ducs nicotine facilitatc avoidance acquisition by enhancing memory con- solidauun or mcrcly by stimulating performance? To answer this qucstion, a scnes of experiments was run on 228 female Sprague-Dawley rats. Animals were divided into groups and trained for 15 one-hour sessions in a discrimi- natcd Icvcrpress avoidance situation with buzzer as a conditioned stimulus. More rats receiving nicotinc, 0.4 mg/kg. t.p• immediately before each one-hour daily session (prescssion) kcached a preset learning criterion than did saline controls, and rats recciving 0.1 mg/kg of nicotine performed better than con- trols but not as well as those receiving the higher nicotine dose. Rats given similar duscs of quartcrnary nicotine bismcthiodidc in an identical presession cxpcrimcnt did not learn more proficiently than controls. Other groups of rats were given four doses of nicotine in photocell activity cages and the doses of nicotine which facihtatcd avoidance acquisition depressed spontatrcous motor activity. Later studies in which rats were given 0.1 and 0.4 mg/kg nicotine or saline i.p. immcdiately after each session (pustsession) showed that the drug also facilitates avoidance acquisition by this method. Finally, rats were again tratned with prescssion nicotinc, but saline was substituted for nicotine begin- nmg on session 16. No drug dissociation effect was seen. Thus the results of thcsc studics suggest strongly that small doses of nicotine permanently facilitate the consolidating neural memory trace and do not enhance avoidanee merely by stimulating pcr(ormancc. t:ricktun, C. K. P.ychuphurmaculnpu (Uarf.) 22:317-368, 1971. F'rum thc Ikpartcncnt of 1'harmaculogy and Tuxicology, University of Kansas Schwl of I'harrnacy, Lawrcncc. CHOLIN1:ItGIC Mf?CIfANISMS AND AVOIDANCE BEHAVIOR ACQUISITION: EFFtiCIS OF NICOTINE IN MICE Two groups of I S mrlc CF-I strain micc were given intrapcritoncal in- jcctwns of nicotinc sulfatc (I.Q mg/kg) or an equivalent volume of 0.9% saline. 12 13
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This trcatmenl was followed. 45 minutes lucr, by giving each animal a singlc training trial in an apparatus dcsigncd to providc for the acquisitiun of a paa..ivc avoidance responsc, consisting of toot-shock given whcn a moruc stcppcd through a holc connecting a small vcstrbulc to a large darkcncd chamber. Responsc latency (interval between placcment and entry) was always undcr IS s(conds. A tcsting trial was given 60 minutcs aftcr training. 'I-hcrca(tcr, activc avoidancc Iraining was initiated. Mice treated with nicotinc sul(atc showed, as compared with the saltne-trcatcd controls, a dccrcascd incidence of active avoidance conditioning without cRects upon cithcr passive avoidancc acquisition or escape behavior. The effect of nicotine was to rcduce significantly the ratio of bound:(rcc acctylcholinc in (hc cercbral cortex. This changc was accounted for by a dccrcased content of stored aminc, parucularly in the synaptic vesiclcs, without any change in the turnover of thc "frcc" storage pool. A difference in the ratio of brain acetylcholinc storage pools accounting for different modes of avoidancc is suggested. Essman, W. B., and Essman, S. G. PsychofoyicaJ Rrporrs 29(3):987-993, 1971. Orhrr au ppurr: National Institutes of Hcalth. From the fkpartrncnt of Psychology, Queens College of the City University of New York, Flushing. N.Y. CHANGES IN CHOLINERGIC ACTIVITY AND AVOIDANCE BElfAV1OR BY NICOTINE IN DIFFERENTIALLY HOUSED MICE The behavioral and cholincrgic cffccls of the drug, nicotine, appear to hc dependent upon cndogcnous changes conferred by differential housing. In this cxpcriauntal study, mice houud either in isolation or in groups of five for 165 days were treated acutely with either nicotine sulfate (1.0 mg/kg) or saline prior to being trained to acquire a passive avoidance respon.c. Isolated nucc showed markedly reduced passive avoidance lcarning. but with nicotine trcat- tnent thcir avoidance acquisition was comparablc with that Of group-houscd animals. In the casc of the lartcr, nicotine did not affect avoidance acquisition rate. Active avoidance training, given following passive avoidance tcsting, indicated significantly impaired learning of the avoidance among isolated micc, without any effect upon escape from footshock. Nicotinc-trcatcd, isolated animals showcd a significantly greater incidcnce of active avoidance behavior, but group-houscd, nicotine-trcated mice showcd appreciably reduced active avoidance responding. Parallel studies of accrylchoGnc poads in the cerebral corux of ddlcrentially housed, drug-trcated mice indicated that isolation kd to a significant decrease in the ratio of 'bouruf':'frce' acctylcholinc; nicotine rtcatmcnt led to an ekvation Of this ratio among isolated mice, hut group- housed animals given nicotine showed a significantly decreased ratio. The in- tctaction of differential housing with drug trcatment may scrvc as a model from which cholincrgic mcchanisms related to avoidance behavior may bc viewed. F_uman, W. B. lnrrrnaaonal Iournal uJ Nruroscirncr 2:199-206, 1971. Other aupport: National Institutes of Hcalth. Frum the 1)epartmcnt of Psychology, Queens College of the City University of New York, Flushing. N.Y. NI('l>IIN1: ('IIANGI ti IN I'.1iG AND 13EHAVIOR AFfF.K 1N7KAVIiNOU.I INFUSION IN AWAKE UNKESIRAINED CATS l hrs rnvcsuigauun was undertaken in order to determine whether intra- vcnuus (i.v.) mlusiun o( small doscs of nicotine would inducc in the awakc, unrestraincd cat, a similar phase of inactivity and EEG hypcrsynchrony as has been rclwrtcd fur slccping cais. I he authors showed that- in the awake animals, i.v. in(tuion of 10 ~,g/kg of nicotine usually lcd to an initial hchavioral arousal and IiLG dcsynchronizauon in the cortex and subcortex. The hippocampus cxhihited thcta rhythm, and the amygdala an increase in rhythmic burst activity. Protractcd immob4dy with hypcrsynchrony in cortex and thalamus, and high voltage slow wavc activity in hippocampus and amygdala followcd. Episodes of slow wave and KEM slcep were often noted. A 30-50 pg/kg dose of nicotine produced similar behavioral changes with increased ckctrocorticogram (ECoG) dcsynchronization or flattcning. A 100 r,g/kg dose elicited marked changes in behavior and somctimcs temporary ECoCG silence. Thus, they conclude, nicotine in(uuon Icd to an initial phase of behavioral and EEG arousal, followed by a phase of protracted immobihty and EGG hypcrsynchronization indicating seda- Uun. According to thcu invcstigarurs, the period of hippocampal theta rhythm and inctcascd amygrlala burst activity appcars to be characteristic tor the initial action of nicotinc. Sladnicki, S. W. and Scharppi, U. 11. .1rrbivrs lnrrrnarionulrs dr Yhurmarodynanrir rr dr Thrrapic 197(1):72-85, 1972. Frrnn the Mason Rcscarch Institute. Worcestcr, Mass. II. 5tudics Relufrd to CarcinoRencsis HISIOLOGY AND 111.TRASTRUCTURE OF CULTURED HUMAN TUMOR CI:LI.S EXPOSED 10 ANTISERA TO Tf1E NFRVE GROWTH FACTOR Various cell changes described in this paper appear to represcnt a specific type of ancigcn-antrtxKly injury. In the work reported here, several dilutions of anti-ncrvc growth factor serum (rom rabbits or horses immunized with purified nerve growth factor were added to rapidly growing. 48-hr. monolaycr cultures of a human ghohlasloma (C356) and Of an H. Ep. 2 cell line. Cytotoxic effects. beginning two hours later and usually progressing to complcte cell death at 17 hours, were studicd in virro in stained covcrslip preparations and by electron microscopy. Amtscrum made in the horse had about onc-hf,h the cytoloxic activity shown by :uttucruru madc in the rabbit. H. Ep. 2 cells were about five umcs as resistant as C3t6 ccllti. In both ccll lines, many of the cclls surviving the antiscrum showed nuclci with gencrally inuct membrartcs, cuntaimng 20 to 50 Of rnnre ruundcd chromrnome-Akc (wdics, suggcsting orophase mitotie activity. ('on,rul nronolaycrs receiving normal rabbit or horse serum were contp:uahlc to the unrrcated controls. The cytoloxic and cytocidal effects are probably of the antigcn-antibudy type. co r C 'dq ~ CJ ~ ® .r~. .t:.. • G ~ @_. _. t g...J 14 IS
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Pinkcrton, f L. Bbagar. 8. Rana, M. W. and Holtwick, S. C'rnirr Ktscurch 31:1483-14g7, 1971. Other supp..rt: National Institulc of Mental Health. From the Departmcnt of Physiology, St. Louis University Schoul u( Mcdicine. St. l.ouis. ANTITUMOR ACTIVITY OF ANTISERUM NERVE GROWTH FACTOR (ANTI-NGF) In an attcmpt to detcrminc whethcr antincrvc growth factor (anti-NGF) aNccts thc growth of chemically induced tunwrs, newborn CF-1 micc wcrc in- joctcd subcutancously with "Wclkonu" anti-NGF for the first five days of life in incrcasmg doscs. Control mice receivcd an cquivalcnt volume of horsc scrum. On day 27, all animals wcre injcclcd subcutaneously with 3 mg of benzu(u)• pyrcnc. Threc days latcr, hal( of the mice which had receivcd horse scnnn wcrc givcn thrcc injcctions of nicotine daily. Results indicated that prcxrcatmcnt of animals with anii-NGF dclaycd the appearance ot tumors induced by bcnzo(a)pyrcne. While in the control micc, tumors began tu appear from day 27 of thc first injection of bcnzo(a)pyrcnc. thcir appearance occurred only at day 90 in immunosympathcctomizcd micc. l.ikcwisc, nicotine prctrcatmenl also dclaycd the appearance of tumor until day 100. Both these treatments, i.c., anti- NGF and nicotine, not only delayed the appearance of tumors but also causcd lower incidence of tumors. While nicotine trcatmcnt dclaycd thc first appcar- ance of tumor and reduced the pcrccnugc of incidence of tumors, it caused significant increase in the averagc wcight of a tumor. Bhagar, 8.. and Rana, M. W. Proceedings of the Socicty for Experimcntal Biology and Medicine 13g(3):9g3- 9g4, 1971. Other support; National Institute of Mcntal Health. From thc Department of Physiology, St. Louis Univcrsity School of Mcdicinc, St. Louis. CHEMICAL CARCINOGENESIS IN SYRIAN HAMSTERS This is an cxtensive revicw of the papcrs on chemical carcinogcncsts in Syrian hamsters (Mojocricrrus aurarus uururus) which have been publuhcd since 1966, when thc ]ast comprehensive onc on thc subject appeared. For the purpose of his discussion, the author dividcs the various papers into scveral groups classified according to the systems involved in addition to two other catcgorics, check pouch tumors and immunology of chcmically-induccd ncu- plasms. In thc section cntit)cd "Gcnctics as a Modifying Factor in Chemical Carcitwgcncsis in Syrian Hamsten," the author also dcscribcs some hitherto unpublishcd matcrial ot his own demonsgrating that gcnctrc factors determtnc the response of hanssters to subcutancous injcctions of 9, 10.lmtcthyl-l, 2- tscnzamhraccrsc (DMBA) (Cancer Res. 32:3G0-366, 1972). Ilc cuncludcs that since, in hu oprnion, the significant role of genetic facturs in ccrtam types of chemical carcinogcncsis in Syrian hanistcn has been ckarly dcnanstratcd, it is imponant to use inbred lines ot thtir hybrids in such work. 16 !(umbrugcr, F. (Hiu Kcsruril Cunsulrunt.i, lnc.) In Hurnburgcr F. (cd.): puthufuA:y u/ thc Syriun flurnitrr. Prnx•rr.u in E.cpari- rnintut 7 untur Kisr,uch, Vol. 16, Bascl: S. Kargcr, 1972, pp 152-175. Othrr suppurt: U. S. 1'ublic Health Servicc, Virginia and D. K. l.udwig I-aundatrun, Fannie E. Rtppcl Foundation, and American Medical Association --liducatiun and Research Foundation. From the Bio-Kcscarch Institutc, Cambridgc, Mass. TH1: DEAF DEX1 RAN:F'OI.YRIBOINOSINATE-POI.YRIBOCYTIDYI.ATE COMI'LIiX: 1')fYSICAI. PKOI'ER7lES AND INTERFERON INDUCTION Tu be able to delineate the mechanism of interferon cnhanccnscnt, studies wcre madc of thc physicochcmical and biological properties of the rl•rC-DEAE dcxtran complcx. f:x{xxrimcnls were carricd out on human neonatal cclls and niuusc 1. ccll% which weic brown as norwlaycrs in MEM containing 15% (v/v) fctal cal( scnum and incubated at 37° in a 5% CO: atmosphere. Bovine vesicular stomaUUS virus (VSV) was harvested front VSV-infcctcd mouse cmbryo cclls. 'so assay (ur intcrlcron induction, cclls were exposcd to rl•rC-DEAE dcxtran complexcs in solutron A for one hour at 2' or 37"; thcn thc cclls were washcd and incubated in fresh medium for 18 hours. A series of tests was then run. In summary, the results of thcsc cxpcrimcnts suggcst that DEAE dcxtnn )w(cntiatcs inacrfcron production by delivering more of the rI•rC complex to the triggering site. It does so by increasing the uptake of rl•rC, through the aggrcgation clicct, and by rcndcring it less susceptible to cndonuckascs. 1'uha, 1'. M. and Curtur, (V. A. Virology 45(3):777-7g1, 1971. Othcr support: Amcrican Cancer S.xicty, Maryland Division. From thc Dcpartmcnts of Medicine and Microbiology. The Johns Hopkins University School of Mcdicinc, Baltimore. SIREI'1'OVAKICINS INHIBIT FOCUS FORMATION BY MSV (MLV) COMPLEX In an attempt to study DNA polymcrasc inhibition, thc cfCccts of s(rcpto- varicins were tcstcd on the cllicicncy of Iransformation by Moloncy murinc \arcoma-IeUkacml:r cUmplCx (MSV) in virro. Also checked were the abilities of the strcptovaricm complex. as well as the purified srreptovaricins (A and D), to alwrt tran.lurmatiun of Balh/3T3 cells by MSV. For these studies, cc11s were grown in the presence of increasing concentrations of antibiotic until ccll growth rates were uh.crvcd to slow. Those strcptovaricins (complex and D), which arc acuvc tnhibhun of the MLV DNA polymerasc in cell-frcc mixtures, blockcd thc formatron of MSV-induccd foci at concentrations which did not impau thc diviston rate of normal cclls. In the case of complex and D, focus formation w:.s hl.k:kcd at cunccntrations o( antibiotic about four-(old less than that which dccrcaw ccll growth. In contrast, the struclurally related macrolidc A, which lacks inhtbuury action on the viral cnzymc(s), was inactive in tissue culturc. 'I hc..r rc%ult% suggcst that thc clfccts on blockade of transformation arc duc lo a sclcctrvc inhibition of KNA-dclxndent DNA pulynxrasc because inhibi- 17

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