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ANDREWS OFFICE PRODUCTS CAPITOL HEIGHTS, MD (K)

1979 REPORT o/ THE COUNCO. FO7t TOBACCO RF.SEARCH-U.9.A., I.e. TEE COUN(71. FOR T()BA(:CO RE3EAR(71-11.9.A., lat. l 10 F...1 59t6 3eree1, New Y.ric, N.Y. 10022

SCIFIV'17FIC ADVISORY BOARD to The Council for Tobacco Re.aucb-U.S.A., Inc. as ot December 31. 1979 SHFII)ON C. SOMMERS, M.D., Cho"sr Dlrenor of Laboratorle.r, Ltaos Hill Hospital Cpnkd Pro/estor of Pathology Colkge of Ph)sician. t Surteoas of Columbia University New Yort, New York RICHARD J. BtNO, M.D. Dlrertor of Crdlolosy and Introrwao/ Afedklne Huntington Memorial Ho.pital, Paadeea, Catitorda Professor of Medklne Unlrenity of Soutl+er. CalYornia Scbool of Medicine [.o. AngeJea, California JOSFPII D. FELDMAN. M.D. Head, Department of leununopatbolop Seripp C1mie and Research Foundation La 1 _olla, Cati(ornia iVQ.UAM U. GARDNER. Pn.D. Sdent!/ic Dirertor, The Council for Tobacco Research-U.S.A., Inc. E.1C. Hrnt ProJcsavr of Anaton+y (en.rritut) Yab Unireniq School of Medicine New Haren, Conoecticut ROBERT J. HUEBNER, M.D. Chfe/, laboratory of RNA Tumor Vinues National Cancerr Institute Betbe.d., Maryland LFrON O. JACOBSON, M.D. loseph Retentteln Professor of Biological Scknres icago Cbkambiooi. HENRY T. LYNCH, M.D. Professor and ClwJrnwn Department of Preventive Medicine and Public Heatth Crel~too Uoivenity School of Medicine Om a, Nebraaka HANS MEIER, D.V.M.. Dr. Med. Vet., M.R.S.H. Senior StaO Scknttrt Tbe Jackson IAboratory Bar Harbor, Maloe JOHN P. WYATT, MD. Tlrector obaoco and Health Research Institute University of Kentucky Les)nSton, KeatucJ:y 9eleetlfe Staa./ T`. Cw.dl WILLJAM U. OARDNP.R, Pa.D. Scientific Dbrelw ROBERT C. HOCKE7T, Pe.D. Research Director DONALD H. FORD, Ptt.D. VINCF.NT F. USANT4. D.MD. Associate Research Director Assodate Reserc h Director DAVID SfONB, Pa.D. As.ociate ResrrdY DLMctor

CONTENTS Introduction . . . . . . . . . . . . . . . . . . 7 Abatracta of Reporta . . . . . . . . . . . . . . . . 9 Caocer-Related Studies . . . . . . . . . . . . . 9 The Respiratory System . . . . . . . . . . . . 22 1leart and Circulation . . . . . . . . . . . . . 32 Neuropharmaoolop and Phrtiobp . . . . . . . . . 39 Pharmacology .ed BiothemLtry . . . . . . . . . . 49 Immunology and Adaptive Mechanisms . . . . . . . 67 Epidemioio8,y . . . . . . . . . . . . . . . . 78 Active Projects . . . . . . . . . . . . . . . . . . 83 Completed Projects . . . . . . . . . . . . . . . . 92 Itsde: of Priscip.l Investigators . . . . . . . . . . . . !03 Index of Scnior Authors . . . . . . . . . . . . . . 104 i I / JOHN P. WYA7T n 916.1980 lohn P. Wyatt, a member of the SckntiAc Advisory Board for aeven years. died January 22. A distinguished pathologist and specialist in pul- monary disorders, he had heew, sinoe 1974. Director of the Tobacco and Health Research Institute at the Univasity of Leaington. Kentucky. Before that he was Professor and Head of the Department of Pathology .t the University of Manitoba Faculty of Medicine, Winnipeg, Canada. lack Wyatt was a modern version of the Renaissance man, aervin8.ocieq well as a physician and researcher at the same lime that he explored (or truths in literature and the arts. He frequently enlivened Advisory Board meetinp with quotationf from literary {re.u that struck directly to the core of the matter under discussion. Articulate, witty, gregarious. ).ct Wyatt dwars sought to Improve the Imperfect world of scknoe. He took panicui.rly to bean his responsibiliy as a member of tha Scknti6e Ad- visory Board and arove in thN body, in his owo institution and el.ewhert, to advance acieati/k knowledge and better the human conditloa !1e wiY be deeply missed.

Introduction The Council /or Tobacco Research continued ouriaQ 1979 to pravide L oancial support (or independent scientists who are aeeking to add to knowledr of canur, cardiovascular diseases and chronic pulmoaary ai4nema. The tact is that the etiology of ttrese map' ~ailmenb r~emaias a mystery despite the vast and evertxpaadics researcb cAort (or . solution. Coo.ideries their compleaNy, there should be no doubt that the aoswer, or amwer., wiY be slow in comind asd will be b uod on discoveries by researchers is masy diecr- tot disciplines. The (acton o[ genetic makeup aad aging are important eoosiderasior is the scientific effort to determine the puhoRenesis of these ailments. Aiso ies- portaot are the recognition and examination of the impact m.de by the toul environment of our iodustrialized society. The areases and strairr of livinL exacerbated by worsening economic tww- ditions and international crises, are being studied more closely for their porr sibk role in body processes and d'ae..e accurretsce. The overall situatioe, /rom a scicatiik viewpoint, is immensely ooesplen, and one cannot expect a simple or easy solution. The Couocil, however, r.- mains optimistic that answers wiN be found eventually; so too ar, other &raM- ing agencies as well as aientists invdved in the research. The Council believes that new ideas and fresh approaches to old but still good ideas will continue to come from the world of scienee. l hat this is true ir aece in the number of requests (or research aupport that The ('ouncjl receivea rc11u- lariy. The ('ouncil remains dedicated to helping support independent isvesd- gauas in their search (or definitive answers to eancer, heart disease and chroaic pulmonary ailments. 7 he following section contains abstracts of research reports published durins 1979 by rccipientc of Couqeil support. 7

Abstracts of Reports Following are aburacts, approved by the autlwri, of reports on oenr research acknowledging support /rom The Council that have appeared ie .cka lifk journala since publication of the 1978 Report. The oame of tho recipket ia in Nalics. The abstracts are grouped under these headinp: 1. Cancer-Related Studie., 11. The Respiratory System. 111. Ileart and Circulation. IV. Neuropharmacolory and Physiob`y. V. phatmacolo6y and Biochemistry, V1. Immunology aod Adaptive Mechanisms, VII. Epidemiology. I. (;ancer-Re/ated S'tudIe. BIOLOGICAL ACTIVf3Y OP TOBACCO SMOKE AND TOBACCO SMOKE-RELATED CIIEMICAIS There are three basic questions concerning the role of tobacco amoke IN human cancer susceptibility: (1) Does tobacco amoke contain aubataooa that interact with the microeomal munooayjenaset (2) I/ ao, what are some of Uw in vivo and in vitro eflectsT (3) Are such eQecb more hmtdoua for urtais individuals or tissues than for others? As ahows here, eaposure to w6ole cle- ueue smoke Irom relerenee cigarettes promptly, but rather weakly, ioducea murine pulmonary tnkro.amal matoosy6enase activity (peak activity at 6 hours; about 2-told). This can be detected with Ihe uae of benzo(a)pyrem or ethoayresorulln as suburates, and is inhibited by cyclohesimide or actiaomycis D. 7 hese smoke-induced incrcnes. Aowever, are not umequivocally linked to the Ah locus as they are when caused by the Intratrache.l admini.uatioa of 3-methykholanthrene. Both whole amoke condenaa/e and its /ractiom ca.: (1) induce pulmonary monoosygenase activity; (2) Mhibit btnto(a)pyrene metab- olism in ritro; (3) be metabolized to subMances muta6enie to SdtnoweBa typAl.nrrirm (TA133t or TA9e); (4) transform C311 IOT% oelb in vltro; and (5) inlensi(y the carcinogenicity of benzo(a)pyrene ht murina lwsue. ro- leatially important ia the observalion that while hepatic tis.ue has the capacity to activale whole cigarette smoke condensate to mutagenic lorma !n v/t% murine pulmonary Iiuue docs nut seem lo have Ihis capacity. Although murina hutg homottenales posse.s slgnilkanl Allil activity and can metaboliae AAa- toain 8,, 2•aminolluorene and 7,tdihydro•7,ldihydroaybenro(a)pyrena to mu• tagenic aubstances. they show only weak activity with the ptommutcn, 6- aminochrysene and no activity with 2A1 cigarette smoke eoodeoaale. Macus- sioa of these results centers on the concept that whole cigarette .moke may be both a potential "iniliator" and it "ptomotor' of lung cancer in mice, and on the notion that the lalter may be the most important determinant of cancer risk. Kouti, R. E. et d. (Alicro6iofoticd Auociatti) and flenrdict, W. F. Environmentd Ilealth lertpertiver 29:63•69, 1979. From the neparament of Biochemical Onedo6Y, Microbiological Associatn, Inc, Bethesda. Md., and Childtcn's 1lospital of Los AnIleks. 9

INHALATION BIOASSAY CHEMISTRY-WALTON HORIZONTAL SMOKING MACHINE FOR INHAl.Al1ON EXPOSURE OF RODEN fS TO CIOARETTE SMOKE Until now, esperimenlal studies pertaining to tobacco smoke carcino- genesis have been hampered by the lack of a convenient, readily vailabk and wcll-deRned system capable of producing smoke for inhalation by (est animals under standardized conditions. TM Wallon Horizontal Smoking Machine ( WIISM ), a commercially available derioa designed to expose as rnany as 20 mice to the smoke of a single citurtle, way provide the answer. Sfudies indi- cate that this machine produces a uniform smoke aeroaol of predictable con- centration with both high and low nicounctontent cigarettes. The aerosol's composition was appropriate to both types of cigarettes: those with a high nicotine conteot each delivered 40 mg total particulate ma/ler, 2 4 mE nico- Iine, aml 17 cc carbon monoxide; thosa with a low nicotine cnnt:nl yielded 30 mg particulate matter, 0.3 mg nicoUna, and 17 cc carbon munoxide. For this atudy, two diRerent qrains of mice. C378L and DBA/2Bd, were exposed in the WNSM. In spite of limitations inherent to a system exposing animals to standing smoke, the WHSM oQers the animal a uniform quaotity of smoke whose concentration and composition (all within those predicted ky standard analytical smoking. The period of exposure is characterized by a~kcrease in the concenlrations of particulate matter aod asost gas phase constiwents, and by an increase in particle size and carbon dioxide concentration in the exposure chamber. Guerin, M. ft. er al (Union Carbide) Journal of the Nariona! Cancer lntrirWe 67(2) :411-41E, 1979. OtAer.upport: U. S. Department of Energy. From the Analytical Chemistry Division, Oak Ridge National Laboratory, Oak Ridge, Tena_ ARYL HYDROCARBON NYDROXYLASE INDUCTION IN MITOGEN- STIMULATED LYMPHOCYTES BY BEN2ANTIIRACENE OR CIGARETTE TARS ADSORBED TO ASBESTOS FIBERS Soma chemical and physical properties of amosile asbestos (AS) fibers were examined here using Ihe human lymphocyte aryl hydrocarbon hydroi- ylase (AHt1) enzyme system as an inveqigative tool. When cultures of mito- gen-stimulaled lymphocytes were Incubated with AS, an increase in Atilt activity was noted. A much grealer increase in enzyme aclivrty followed addi- lion of the inducen benzanthracene (BA) or cigarette lars (CI ) to cell cut- tures. Significant enzyme induction also occurred when AS fibers were 8rst preincubaled with CT or BA, washed with acetone, then added to lymphocyte cultures. lhis increase in Atilt activity was not as great, however, as the in- durliun uMervcd when BA or CT was added to cell cultures. Overall, the cnrynsc .clr.rly in cells rncubated with (T preuealeJ AS par/rcks was not as Rrs.r .% she aar...ry in ('1 rnJrrceJ IyrnpMrcyres ur nr lymphrnyla 14- whrah t 1•u.l \\ ,.crr .u..ulr.nc.ruslr aJJeJ Ihese rc.ulls mJrC.le 1h./ puly:yclie a ru.ru hsc6- %.sh .. NA .nJ cumirrnrnl% ui ( 1, cau Ire ad.urhed i to AS parlicka, which then function as carriers for the transport of these com- pounds into cells. McLemore, T. L., Jenkina, W. T., Arnorr, M. S., and Wray. N. P. Canrir Lerrerr 7(2,3):171-177, 1979. Other lupporer Veterans Administration Hospital. From the Veterans Administration Hospital; Departmcnt of Biology. "ibe Uai- versity of Texas System Cancer Center; M.D. Anderson Hospital and Tunar Ins1i1u1e, and the Dep.rtment of Medicine, Baylor College of Medicina, Ilouston. INDUCTION OF ARYL HYDROCARBON HYDROXYI.ASE IN HUMAN PERIPIIERAI. BLOOD LYMPH(X:Y7E3 BY CIIRYSENE Many polycyclic aromalk hydrourbons (PAIIa) Induce aryl hydrocarbon hydroxylase (AIIH) in human lisasres. This report describes the rok of chrys- ene-an environmental contaminant formed aa a result of the pyrocondeaaa- tioa of various cornpounds, including tobacco products-as an inducer of AHH in human lymphocytes in vitro. Lymphocytes obtained (rom healthy human subjects were cultured for •tl hours, then incubated with chrysene i. rcetone for an additional 24 hours. The chrysene-induced cella showed a aill- nifkant increase in cnzyme induction when compared to controls incubated with acetone alone. Preliminary observations suggest that the time course (or chrysene induction of Atilt is similar to that of benzanthracene (BA) and 7-nxlhykholanthrene (3-MC) induclion, the maximum being reached linearly after 24 hours of incubation. Other s/udin with pregnant rats suggest that chrysene also increases AHH activity !w r,iro. Tirc present study, moreo.er, indicates inlerindividuat variation in ANN inductibility with ehryaene, but this is commensurate with that obtained with SA or 3-MC. While there Is a theoretical relationship between Atilt induction and chemical earciaoRenesis, the connection between carcinogen metabolism and cancer susceptibility is man has not been established. Consequently, reports wch as these might help to determine the role of enzyme induction in Iha susceptibilily to diaease caused by exogenous agents. Although Ihe meubolilea of chrysene produced by At1Fl are only weak carcinogns. the abundance of this hydrocarbon ia cigarette smoke might have an idirect but significant rok in the amplifkaliow of the Atilt system in the lungs of cigarette unokers. Snodgrass. D. R., Mcl.emore, T. 1.., Marshall, M. V., Wray. N. P. Cantrell. E. T., Busbee, (). 1.., and Arnar, M. S. (MarNw, R. R.) Cancer [.erters 7 : )1)-)1 e, 1979. Other .o'portr lhe Welch Foundation, Nalional Institutes of Health, Amer- ican Cancer Society, and the Veterans Administration Hospital, HouMon. From the Department of Biological Sciences, North Texas Slate University. Denton; [kpanment of Pharmacology, Teaas College of Oaleopathic Medicine, For/ Worth; /kpartmenl of Environmental Biology. l)nivenity of Tesas, M D. Anderson Hosprul and Tumor Institute; (kpanment of Medicine, Baylor C'olkge of Medicine; and Veterans Administration Ilospilal, Houston. 11 10

INDUCf1ON, INIIIBITION, AND SOME ENZYMOLOGICAL PROPI:RTIFS OF ARYI. IIYDKO('ARBON HYDROXYLASE IN FRL'SII MITOGEN-AClIVAT1:D HUMAN LYMPHOCYTES This report describes the induction and Inhibition speciflcities and some other enrymologic properties of lymphocytic aryl hydrocarbon hydroaylase (AHHI• Basal and/or polycyclic hydrocarbon (PAII)-induced AlitJ in mito- ten-activated cultured lymphocytes from healthy donors were used to study these properties. Several compousds with an Inducibility ralio > 2.0 were found among the 24 chemicab lested: 2,I,7,S-telracMorodibenzo-o-dioain (TCDD), dibenz(a,li)anlhracene (D<!A), benz(e)anthracene, 7-methykhot- anthrene, P -naphthoflavone, chokcakiltrol, and tx.-isoproterenol. The Rrs1 four, Iisled in descending order of potency, were Ihe mwt vigorous inducen. lhcse appear to activate the common penelitally determined factors involved in Alllt induction, suggesting that they could be used Inlerchanlieably. EAects ranging from inhibition to mild inducliorr were obtained with other substances: 1-bromoffavone, .-naphthofiavont, chrysene, p.,*-1,1,1-Irichbro-2,2-bis ( p<hloro- phenyl)elhane, 7,12dimelhylbewnthrateee, pyrene, o,z-norepinephrine, bcnzo(r)pyrene, lindane, n octylamine, Ie.toueroee, 2.3diphenybsar.ok, 170estradiol, mclyrapone, and phenobarbital. The inhibilon were ranked In descending order of potency as follows: mr-napMhoflavone, p-naphlloflavone, 2dielhylanainoe/hyl-2,2-diphenylvakrak, nsetyrapone, 1,1,1-Iricholoropropene oaide, and cyclohesene ozide. Depending on their concentratioe, the Iaa1 four produced moderate inhibition to moderate stimulation. However, the inhibitioe pattern for the basal and the PAH-induced AIIH was Indistirguishabk. Both the half-life of the enzyme during cell culture and the K. values of A/IH in uninduceJ and PAII-induced cells were similar as well. Based on this and other evidence, it can be concluded that: (1) 'ICDD. DBA, benz(.)anlhracene, and .l-methykholanlhrene have a common mechanism of action whereby only the degree of AHH induction produced dt8ers; and (2) basal and induced AHII are qualitatively similar and only quantitatively different in comparabk unin- duced and PAH-induced cells. Gurtoo. H. L. ct ol. Cancrr RrsrarcA )9(11) :1620-1629, 1979. Other aupp.rtr National Cancer Institute. From the Departments of Eaperimental Therapeutics. Molecular BirJoty. and Immunobpr, Roswell Park Memorial Institute. New York Slale Department of I/ealth, suflalo. ARY1. HYDR(X'ARBON HYDROXYLASL' IN A STABLE HUMAN B I.YMPII()CYl F. ('EI.I. LINN, RPMI-17H0, CUL7URED IN TIIE ABSENCE OF MI t(X3ENS K/'M1 170e is an imorunoRlnhulin synlhesiring B-lymphncyle cell line that has shown hish basa and indrKrbk aryl hydrocarbon hydroaylasc (/1/11t) at tivily in the ab.rne c rtl nutuRcmc preslimulation Durin6 the course of the c.l,~nruts ir/NUi.J hrec. .ri,aut p.ramcters influcncmg the enzyme activity i and inducibilily were studied and optimized. This allowed altlainmeett of enzyme levels and inducibility ratios comparable to those previously reported for fresh milosen-aclivaled human lymphocytes. However. Ihio cell line haa several advantages over fresh lymphocytes: (a) high AIIlI aclivNy in the absence of milogen preslimulation; (b) availability in unlimited quantities le the absence of donors; and (c) the consistency gained by using a ain6le huma. cell line free from seasonal variation of donors. It'seems, therefore, that this system could provide a good esperimental tool for studies on human mised- tunction osygenases. In addition, using cells cultured under oplimal,conditions and induced optimally with dibeoz(.,h)anahraccne. the melabolisrn of bee- zo(a)-pyrene was analyzed by high-pressure liquid chromatography. The melabolile profik produced by these cells had severd similarities with Ihosa produced by mitogen-uimulated, abort-term cultured human lymphocytes de- rived from fresh blood. Freedman, 11. 1., Gwroo, Il. L.. Mieowada, l., PalSee. S., and Vaught, l. 0. Cwscrr RerearcA )9(11) :1605-1611, 1979. ' Other.rpportr National Cancer Institute. Front the Departments of Eaperimenlal Therapeulics, Molecular Sioloq aed Immunology, Roswell Park Mernorial Instilute, New York State Department of Health, Buffalo. INDUCTION, INIIISITION, AND eIOLOOICAL PROPERTIES OP ARYL HYDROCARBON HYDROXYLASE IN A STABLE HUMAN B-LYMPHOCYTE CELL LINE. RPMI-178t The present paper reports on the ieductbn ud Inhibition by .atious chemicals of aryl hydrocarbon hydrosylaae (AHH) iw a slabk, imnwteolog. kally de6ned (ISM-yiek/ing) S-lymphocyle line (RPMI•17llU), as well aa ae the kinetic and biological properties of the ewzyme In these cells. This pr- ticular line had previously beee selected as a biochemical ruodel for AHII .ludies mainly because of Its hi6h enzyme activity ie the absence of mitopeos. In the dose ranges tested and on a molar baia, the inducers, in deaoeodie6 order of polency, were: 2,),7,!-telrachlorodibenzo-p-diotin, dibenz(.,A)aa- thracene (DBA), l-methykholanthrene, benzo(.)pyrene, and 1,2-beezanthra- cene. Included among the potemial inducers that paradoaically lowered basal AIIlI, were 7,12-dimethylbenzan(hracene, 2,Shcphenylosazok, and chryacee. Under optimal culture curdilions, induc/ion promoted maaimum enzyme activ- itics three- to four-fotd that of batal AIIH. lYm characterhllcs of the 1)sA-In- duced and basal enzymes were virtually idenikal. lhe pat curves were similar for brMh (R.23 optimum), as was inhibilor specificity. Induced and basal enzymes displayed similar half-lives, apparent activation encrgies, optimal tem- peratures. denaturation temperature range, end apparent Kr„ with benzo(.) pyrene. lhe small differences observed in Ihe latter were related to the enzyme concentratarn during incubaion rather )han to its yuai/y. Freedman. 11. 1., Parker, N. 8,, Marinello. A. 1., (irrroo, 11. l.. and Mino- wada, 1. 1) 12

Cancer Rerrarch )9:4612-4619, 1979. Ottier.upportt National Cancer Itatitute. Prom the Departments of Experimental Thenpeutia and Immunology. Roswell Park Memorial Institute, New York State Department of 1(ealth, Buf(ab. CELL SOR i'ER ANALYSIS OF CARCINOGEN METABOLITES IN HUMAN 71SSUES The biochemkal parameters of a betesv~eoaw culture of human (ympho- cytes were eaamined with a f)sortsoeeoe-activated ceN sorler (FACS-11). During culture, the cells were incubaled i. the presence of benz(a)an:hracene (BA) to induce the enzyme system that metabolizes carcinogens. Direct assay of carcinogen metabolism was achieved by measuring the phenolic meubolites of cells e.posed to benso(a)pyrene (BP). According to single cell measure- ments. BP metabolism was grealer in tbe larser cells than in Ihe smaller ones. In the cells exposed to BA during cultura, the entyme aclivity also was greater (or a given cell size. In some of the studiea, dabk celb were Ont sorted out sccordind to size nd the wbpopulations were tusayed (or the o-deethylation of eihosyresorufln, a compound that measures eylochrome P-44E1 activ:ty more specifically. In corroboration with the direct metuuremenu, the larger cells demonstrated higher en:yme activity levels thas 1he smaller ones. The technique of sorting before enzyme assay. as described here, can be applied to other tiswes, as well as to other cell types asd enzyme systems that cannot otherwise be measured directly. Tyrer, lf. W., Adams, L. A.. Tdlany, S. M., O'Connell. J. P., and Cantrei(, E. T. The /ownd of !llytocheminry and Cytochemirtry 27(1) • 30N-S 11, 1979. From the Becton, Dickinson Research Center. Research Triangle Park. North Carolina; Tesas College of Osleopathic Medicine. Fort Worth; and Division of Cytopslhdop, 1)epartment of Pathobp, The Johns Hopkins University School of Medicine, Baltimore. ACTIVITIES OF POLYCYCLIC HYDROCARBON ACTIVATING AND INACTIVATINO ENZYMES IN HUMAN LUNGS OF SMOKF.RS, NON-SMOKERS, LUNGCANCER AND NON~CANCER PATIENiS Polycydk hydrocarbons (PAH), which are present everywhere, eaert lheir adverse effects only afler metabolic activation. The mkrowmnl nwno- o,ygtnase system (MO) catalyses tM osidaliw biotrans(ormation of asreat number of endo6enous and exogenous compounds and ia also rtsponstbk for the formation of eiectrophilically reactive eposides from PAII. Concentrations within the ceUs of Ihese reactive epo.idet, which are suspected ultimate car- cinoRens, may be conerolkd no1 only by the MO aclivitits, but alur by the eposrde•metabuli:ing epoaide bydratase (PH) and tlutalhione-Suarrs(erase ((iF) ectivilies. Since such dats did not seem to eaisl, the epoaide (orming and metaboli:ind enzyme activities were meuured in 47 ssmpks from lunp of smukers, nonsri>,.\ers, lung cancer and noneaneer patients and correlated to the rMcurnence rd e.ncer When uand.rd assays were used, no Ml) activity .uulJ f.r dcte.ted m hun..n lunp, alth.>,t6h the very setadrve assays with ben:o(a)pyrene, 7<thoaycoumariq 7-etboayreaoru(in and biphenyl as wtb. strates were used. However, these activities in rat lung mkrosornes used as positive controls were always clearly meawrabk. By ooincub.tios of ral lueg microsomes with human hms microsomes, It could be demorulrated 16at 7- etho.ycoumarin-odedhyl..e activity wu inhsbiled by the presence of hurn.n lung microsomes after preiacubation for 13 minutes. 'Ibes. Bndinp wyeM the presence of  diffusible inhibitor is human hmR microoaees. 1. 16e enzyme activities of human lung aampks, considerable i.lerindi.idwl va,riiatioas were observed. However, no sisa(kant diRereuces (P>0.1) were found wbes enzyme activities in IunRs from notscsaeer patients awd .oa.rnoken ww compared with thow from lung cancer p.lie.ts, suggesting Wy other (acfors may affect differences is lung tumor wuceplibility. Lorens, 1., Schm.asmann, H.. Ohnhaw, E. and OercA, F. Archlver of Toairofoq. Suppl. 2(Meehanitrn of Toxic Action on Soene Tarpt Orpns) :4l1-4l9, 1979. From the Section on Biochemical Plurm.eobQr, Institute of Phann.oo{ogy. University of Main:, Main:, Wes1 Germawy; a.d Inaelspitd Bers, Switzerland. A NEW ASSAY FOR GLUTATHIONE S-TBANSFERASE USINO (•H)- BENZO(A)PYRENE 4,S-OXID6 AS SUBSTRATE. INDUCIBI(.ITY BY VARIOUS CHEMICALS IN DIFFERENT RAT TISSUES COMPARED TO THAT OF ARYL HYDROCARBON HYDROXYLASE AND EPOXIDE HYDRATASE A new assay for glutathione S-tratuferass that uaa (s11)-benrp(a)pysw. 4.5-oxide as a substrate Is described. Compared to previously propo.eA net6ob. this assay offers several adoantapes: (1) it ia easily .od rapidly per(orsnea, requiring only one extraction step and eliminating the usual chromatorphic sep.ratioa of substrate and products; (2) it tNilires as a substrate am s ids metabolically produced from a ea(einodetsie polycyelie aromatic hydrocarbon; and (3) it is performed under optimum conditions for the pll and eo.oes- trations of the two substrates (ghNalhione and epoaide). This assay wr used 1o study the effect on Shnathione S-transferase of compourds kt.ows w induce drug metabolizing enzymes. The activity of gIWathions Stransfetase was oaly slightly aflected by varioua inducers. In this respe.ct, it ia very diRerent from other enzymes inswlved in polycyclic hydrocarbon metabolism: liver epoalde hydralase was very sianifkanty enhanced by phenobarbilal and 16„tyaoo- pregnerwrlon. Iwatmenl, and aryl hydrocarbon hydrosylasa appeared to be very easily arnd strongly induced (n many wgans by several chemkals. 'Ihese results further support the eoMention that in contrast to other en:yaw, glutathione S-transferase itduetion probably has only a minor role in the regulation of polycyclic arumalic hydrocarbons elwninalion. Van ('anlfort, I.. ManU, 1.., (ikkn, I. E.. (Ball, 11. R., aad UcrcA, F. Biochemical Pharmacology 2(1(4 ):4S S•460, 1979. From the I ahoratoire dc ('himk MEdicak, Imtitul de Palholugic, San Tdmaa par I itae, Nelsium; and the Section on BNrchemKal Pharmacolo6y, Institute of Pharnsacology. Universtty of Mainr, Mainr, Wcst ('.ermany. 14 I'S

i i { RI:Dl1C1lUN OF t11:NZO(a)PYRENE MUTAGt:NICITY BY DIIIYDRODIOL UEHYDRO(iENASti Since epoaide hydratase cannot inactivate the dihydrodiol epoaides pro- duced by the environmental carcira8en, benzo(a)pyrerse, the abiLty of a dihydrodiol dehydro8enase to sequester the precursor dihydrodioh an,' thereby to aflord protection from the later, more reaclive, dihydrodiol ept..i•Ics was investigated. To carry out this study, a purified form of the enzyme had to be prepared. The effect of pure dihydrodiol dchydro8enase on llse mula- Senicity of bento(n)pyrene was then studied in the microsomc/Salnwnclla lest. Here two metabolic pathways mainly contribule to the muts8enicily of benro(a)py'ene: (1) activation to bento(a)p)rene-,,s-oaide and (2) drhydro- d'wl epoaides. ihe Salnwnrlla rypAlnrrrlvrw arains TA 98 or TA 1110 are highly susceptible to bulh Sroup" of muta8enie wrelabuliles, whcrea Iisc strain lA 15)7 is much mote susceptible Io the 1,l-oside than to the drhyQrodnA epoaidts. Results showed that when benzo(a)pyreno was activated by liver microsomes from 3 nselhylcholanthrene-trealed mice, addition of pure dihy- drodiul dehydrogenase ekarly reduced the reula8enicily. The effect increased with the duse up to 0.5 units per plate; (urther addition of dihydnAiul de- hydro8enase had no si8nifkant effect. The percentage of the bcn:ol.)pyrene mutarcnicrty which could be removed by dihydrodiol dehydro8enase depended oo the bacterisl strain. With 1A 98, it was 30-609i, and with TA IS37, as eapecled from the lower sensitivity of this strain (ot benzo(e)pyrerse drhydro- dio) eposides, it was smaller. lhus, it was concluded that dihydrodiol de- hydru8enase can protect atainsl muu8enie effects ol benzo(a)pyrene, but that not all the mula8cnic mcuhuhtcs ul bttuo(a)pyrene esn be rensuved by lhis enzyme. Howevcr, the mutagemctty mediated by the drhydrodiu6epuaide pathway opens an intriguing perspecttve for the entire field of polycyclic hydroearbontvoted muls8enicity, carano8enicity and cyloloaicity. (11a11, II. R., Vogel. K., Bentley. P., and Orsch, F. Naturr 277:319-320, 1979. From the Section on Biochemical Pharmacology. Institute of Pharmacolop. University of Mainz, Maioz, West Getmany. MUTAGENICI TY TFSiING OF NITROGENOUS COMPOUNDS AND 1H1iIR N-OXIDIZt:D PRODU('iS USING TRP+ IN E. COU The N-oaidalion of certain aromatk amirses, aromatic ansiJcs and carbamates can produce mula8enic and carcinogenic compounds. Consequenlly, the development of short-term tests designed lo evaluate potential nsYs front new compounds is highly desirable. lhis report describes a spot test ut:lizin8 tryM"phan ( Trp t) reversion In E. coli to detect substances that cause Aase-pair substitWion, a measure of mula8enicity. A variety of nitro8entontainin8 compoundi and their chemically synthesized N-oxidation products was es- amined 1 be N oarnlalion of certain aromatic amines and earb+malcs was assuaiatcd with mutagenic aclivrly, bul that of aliphalie amincs, tertiary sro- matrc annnes. helrnMyclic numrtic amines, and aromalic aosides did nu1 yicld pnMlutts srl;nificandy nwre nnwagcnic than the parent cumprwnds. 16 I i t Pai, V., Bloomfkkl, S. F., lonca, l. and Gorro/, !. W. In: Gorrod, l. W. (ed.): siiolosird Otidatlow o/ Nlnotew. New Yortt: Elsevier/North-Holland Biomedical Press, 1978, pp. 375-382. From the Department of Pharmacy, Chelsea College. Univenity of l.ondon, Londoo. TIIE IN-VIVO N-0XIDATION OF 3-SUBSTITUTED PYRIDINES FOt.1.OWING 111E1R INTRAPERITONE'AL ADMINISTRATION Previous Mudin have shown that several 3-subslitutcd pyridina ara oaidired /n rlno al the pyridyl nilro8e.. ihe in rlro meubulism of pyridiae and of its l•chkxo and 3-methyl derivatives was therefore resaamined Is order to determine the role of N-oaidation In the metabolism of these com- pounds. Following the intraperitoneal admi.istralion of thc parent pyridina, pyridine-Noaide, )-methylpyriditse-N-oaide and 3<hloropyridine-N-oside were detected in the urine of mice, hamslen, rNS. Suinea pip, rabbils, and fcrrcu. N-oaidalion proved to be a quantitatively Important metabolic pathway /or pyridine. The urinary eacrelion of the N-oaide ranged from about 10% o/ the dose adnsinistered in ra/s to about 40% In mice and Suinea pip. With 3chbropyridine and 3-melhylpyridine, the N-oaides accounted for kss Ihan seven % of the administered dose. Mice prelrealed with phersobarbitnne showed increased urinary levels of pyridinc-N-oaide, but 3-nrcthykholanthrene pro- treatment had no noticeable eRect. Damani, 1.. A., Disky, L. 0. and Gorrod, J. W. In: Gorrod, l. W. (ed): eidorkal Oa/datlon o/ Nluotew; New YorT: t:lsevier/North-Ilolland Biornedical Press, 1978, pp. 157-162. From Ihe Deparlment of Pharmacy, Chelsea CoqeBe, University of l.oedo., Londas. SOME FACTORS INVOLVED IN THE N-OXIDATION OF 3. SIIBSTfI UTt:D PYRIDINES BY MICROSOMAL PREPARKiIONS IN Vf7'RO While the uaidalion of nkrNinamide to the N osWe has been demon- slratcd ln r/tro. the N-osidaliun of the pyridyl nitrogen in various drup and c.Hnpounds with this type of amine function has not yet been studied ea- tensively, either in rirro or its riro. There/ore, in order to establish the occurrence of pyridine-N-usidatiun and to establish the en:ymolo8y of Ihe various reactions invulved, a systematic study of hetero-arornatie N-osidation was begun with the use of a series of 3•substituted pyridines as ns.Klet com- pounds. It has prevronsly been shown that oaidation at the pyridyl nitrogen appears to be a Ecncral melalxslie In virro routc /ur several 3-substiluted pyrwlines. lhc elkcts of various factors on the C- and N•oaidation of 3-melhyl- 17

pyridine in virro and on the N-osidatioo of pyridine and 3<hloropyridine are now reported. According to the data, pyridine-Nozidase activitl, is found mainly in the hepatic and pulmonary tissue microsomal fractions of various eaperimcntal animals. The level of enzyme activity is not neceuarily the same In a1I spccies, however, as shown by the fact that it propeaively declined as rabbits, mice, guinea-pip and rats were substituted for hamsters. Ra1a and mice also showed a sea diRereocs. The speciflc kinetic data (K. and V.„) for the N-oaidation of pyridine, 3-methylpyridine and 3-chloropyriline in various species and orgaos are reported. Gorrod, I. W. and Damad, L. A. Xcnob/ork. 9( 4):209-21 fi, 1979. From the Department of Pharmacy, Chelsea College. University of London, London. TNP. EFFECT OF VARIOUS POTENTIAL INHIBITORS, ACTIVATORS AND INDUCERS ON T11E N-0XIDATION OF 3-SUBSTITUTED PYRIDINES IN VITRO In their continuing series of eaperimenta on the Noaidalioa of 3-sub- slituted pyridines, the authors now present evidence indicating that this type of naction le medlated by a cytochroma P-450 aystem, which is different from eytochrome P-446 and from the flavoproteio<ontaining amine ozidase. The Noadation of pyridine, 3-mclhylpyridine and 3<hbropyridine was inhibited by SKF 525A (2-diethylaminoethyl-2,2diphenylvakrNe) and DPEA (2-4- dich{oro-6-phenylphenoayethylamine). These compounds also inhibited Ihe C-oaidatioo of 3-methylpyridine. Various other nitrogenous substrales, io- cludin6 noclylamine, also inhibited the Nosidation of these three pyridines. When microsomal fractions were incubated with NADPH In a carbon mon- oside almosphere, the N-oxidation of aM three substrates and the C-ozidalion of )-methylpyridine were inhibiKd. In additioe, any treatment of microsomes that reduced eytochrome P-430 concomitantly decreased the Nosidation of the pyridines. Pretreatment of rabbils with phenobarbilone signi/lcantly increased N-osidatioa of the pyridines !n virro, while pretreatment with 3-methykholan- threne had no appreciable effect. EsaenliaBy the same results had previously blen obtained with hamsters and guinea pip. Pyridine-Nozidase, therefore, is clearly similar to the group Ila Noaidase rather than to the tertiary amine osidase. Tlase data support the view that factors olher than the subslrate: dissociatioe eonslant, such as nuckophilicity and nitrogen atom hybridizalion, are among thoee that determine the differentiation of the variaus enzymatic N-osidative processes. Gorrod, l. W. and Damani, 1.. A. Xtnobiorica 9(4):219-226, 1979. Frnrn the Ikparrment of Pharmacy, t'helsea College. Univeniiy of l,nndon, I ondon c T71E EFFECTS OF CYTOCIIROME P-430-44t1 INIIIBITORS ON TIIE BINDING OF BENZO(a)PYRFNE AND DERIVATIVES TO DNA UPON MICROSOMAI. ACTIVATION Whik the rok of cytochrome P-430-44tl-dependent oaidalive pathways in the carcino~enicNy o/ benzo(a)pyreoe (BP) is well krrown, the ideetifkuioe of 6-hydrotymelhyl-BP as a carcinogenic metabolite of BP and the aubaeqrreot determination that the arylhydroaymethyl synthetase «actiws in indepeadcsl of cytochrome P430 indicates that oon-cytochrorne P430-44t{ pathwaya may also be involved in the formation of prodmate carcinogens. This study ea- amines the possible role of tae<ytochrome P-J30-448 pathways i. Ihe covalent binding to DNA of BP aod of the 6-aubslituted derivalives of BP, 6-methyl-BP, 6-hydrozymethyl-BP, and 6-formyl-BP. Resulta show that liver microsomal preparuions from rau prelreated with 3-methykholaolhrene I. the presence of NADPH have an increased capacity to form acliw metaboGta (rom BP and from the 6-substituted BP derivativea. Two different typea of cytochrome P-4S0-44t inhibilon..-tupthollevoee and 1-beozylimidazole, show greater than 80% inhibition of the binding of BP to DNA. I. the presena of these inhibilora, 6-hydroaymethyl-BP, 6-methyl-BP and 6-/ormyl-BP show varying degrees of inhibition o( binding to DNA, depending upoa the iabibilor used. Also, it can be seen that potyRuanylie acid is the most eQective wrb- strale for the binding of each activated polyewekar aromatic hydrocarboe; polyadenylic acid and DNA show essentially equivalent binding. Ttr.r.fora, the data presented in this paper clearly demonstrate that microsanal prepara- 3 from methykhdanthreoe-pretrealed rau, in the preaeoce of NADPH, ate 6-substituted derivetiva of BP b InNaboliles which oovalently, bbind lo DNA In the prsance of two diQerent inhibiton of cytochrom. P-430-44t, under coodilions whereby activation of BP prr re is inhibited. T1se metabolic rowea of the activatiow of dre 6-subsUtuted BP derivatives are not yet know.. Liu. W-1. and Sloawe, N. N. Xrnobloaka 9(l):163-171, 1979. OtA.r a.pporet American Caacer Society. From the Department of Biochemistry. Univer/Ny of Teneessee Center for the Health Sciencea, Memphia- EFFE('TS OF A CYTO(.'HROME P-4301NHIBITOR ON COVALENT BINDINO OF (rHIBENZO(a)PYRENE IN NILCEL.L. FRACTIONS Does 1-benzylimidazok (81), a potent inhibitor of eytochrome P-470 hydrosylatioo reactiona, inhibit the covalent binding of ('lllbeezo(.)pyKrM (BP) to both the nuclear and tytoplasrqic fractions of Nil eelh7 Eaperimeou show that incubation of lirst cycle celb with 81 for 24 hours inhibits 70% of the eovaknlly bound cytoplasmic BP. In the nuclear fractioe. BI inhibiu 33% of the BP binding. In the second eyck. BI activity seems to concentrate in the nuckus, since the addition of more BI to the p.eviously treated nuclear (rae- lions does not further increase its inhibitory effect on 8P binding In the cyurplasmic fractions, however, 81 inhibits 47% of the BP binding aad additional 1rcalment causes 70% inhibition. Inlereatintly, there Is evidence, IB 19

according to aoother report, that benr]midazde derivatives covaknlly bind to nuckic acids. Sloane. N. N.. Chu, L. N. and Amo., H. IRCS Medicd Srknce 7:t1, 1979. Other wpportr American Cancer Sockty. From the lkpartment of Microbioloby ud Medh:al Qeoetks, Harvud Medical School, Boston. REACTIVE ANTIBODIES IN THE BRONCHIAL WASIUNOS OP LUNG CANCER PATIENIS Tumor-rcactive snlibodks have bee. wooerfuly isolated from cluales of lung csrcinomas and from their eAusions in the p..t. To esplore the woleat and the specificity ot immunopobulau (rb) in broechial wuhinp, specimem were obtained from 26 patients with various types ot lung eaneer, 11 patienU with non-neoplastic pulmonary diseases, and 10 hmp with no evidence of lung disease. Fspressed as Ib/potasaium ratios, statistically sipniflcanl increases in 1S levels were found in imflammalory diseases and even higher increases In lun6 carcinomas. The iwlation of I6 from bronchial washinp was a:hieved by dissociation of immune completey at low plt, neutralization and subiepuent purification by anion eschange chrornatography. The isolated immunogtobulins were tested in indirect immuno/luorescence against luna cancer cells of various histologic types in tissue cultures and fresh suspemions. Positive cytoplasmic fluorescence was obtained with cells of adenocarcinomas and spuarrwus-ceU earcinomas of the lung but not with normal lung eells It seems, Iher,efore, that together with the infiltralion of lymphocytes .nd plasma eella in the tumor tiaues, the formation of spccdkaily reacli.e antibodies represents a parl of the immune response to the growth of lung cancer. The accessibility of bronchial washings makes the estimation of their lung<ancer-reactive 1S relatively easy and raises the possibility of Ihis measurement's eventual conversion into a screening test. Paluch. E. and IoarliLn, H. L. Inrcrmrlond Journal o/ Cancer 2)(1) :42-16, 1979. From the Departments of Pathology. Columbia University College of Phy- skians i Surgeons and Lenox Hill Hospital, New York. HEREDITARY CANCER: ASCERTAINMENT AND MANAOEMENr Heredilary, cancers and precancerous diseases have several eharacteristica-- early pe of onsN, bilateral cancer oecunesses in paired orbans, increased occurrence of nwhicentric cancer, and segregation patterns consistent with autosumal dominant inheritance-that distinguish them from other cancer patlerna /heu cMractenstKs can be used as familial cancer selection criteria when rdentdkd in sr.larcd patienls and nuclear family cancer clusters Familial muluple . Lnawna~~~~s pA>P^is of the colon (fPC). which is frequently cited as the cla.+wsl poui.,t>pc ol a hereditary cancer-prcdisposing disxttder, is eum- I i ined in this paper. The historical perspective provided by FPC and depicted la a table here provides an excellent example of the advances in cancer epidemiology and characterization of hereditary precascerous discase(s). Receotly, a Seredi- Iary. premelanoaic cutaneous phenotype (characlerised by multiple large moMs, irregularly shaped, reddish-brown to pink ia color, and with evidence o[ pijmaa- tary leakage) has been iden/i0od. This cutasrcan peeeancer phenotype, which is transmitted in a manner consistent with ae autowmal dornieaot moda of b- heritance, has had the acronym FAMMM, for Familial Atypieal Multiple 111otr Melanoma Syndrome, ascribed to it. Multiple endocrine adeootnato.is (MEA) has also been described as a familial entity. The Cancer Rebistry-baseA probrns which is dealt with here has clearly demonstrated t6a man.er in which a carefully obtained famiy (niNOry can lead to the recppiilioo of patients with exceedingly high cancer risks. Lynch, N. T. er d. CA-A Cawrrr Iorrnd /or CUn/c{aws 29(4):216-212, 1979. Other .upport: Amerkao Cancer Society, Nebraska aod Iowa Divisions. From the Department of Proventive Medicine/Public Health and the Oncology Clinic, Creighton University School of Medicine, Omaha. FAMILIAL FACTORS IN BLADDER CARCINOMA An increasingly valid overview of cancer etiolopr has beea made poa. sible by highly accurate verification of cancer p.lhotogy at all anatomk dtea, and by a meticulous recording of peneabtr, associated diseases and k.ows environmental exposures. While carcinoma of the bladder (s the most coremos malignancy of the urinary tract, surprisingly liltk is known about bost factors in such cases. In this sludy, two families prone to transiliona1 cell carciooma of the bladder were investigated. It is reasonable to wume that members ol the subject families may be snore suseeplibk to various espowrea to cu- einogens, a concept that supports a genetie<nviromnenlal interaction hypotheais for cancer etiology. Identification of hibh-risk individuab should prompt spec cilk surveillance programs instituted sibnifkanlly earlier for such person. Ihan for the general population. Such programs should include urinalysis at regular inlervals to detect mkroscopic hemaluria or pyuria and sedimentary tumor cells. In addition, cancer control measures should consider preventive programs aimed al the avoidance of carcinogenic exposures such as eipretlie smoking by high-risk relatives of cancer-affected probands. The etiology of familial bladder cancer may be a complex one, possibly Involving other associated malignancies arn//ur certain other non ncoplaslk ulmenls, in addition to spe- cifk carcinogenic esposures. Detailed reporting of /amilies prone to this dr- order, including cunsideration of all the potentially important associated /aclors, is seriously needed in order to understand it more fully. Lynch, H. T, cr al. The Iournal o/ Urology 122(1):I3/-161, 1979. From the Departments of Preventive Medicioe/Public Hedth, Urology and Biochemistry. C'rcighton University School of Medicine, Omaha. 21 • 2O

COORDINACY OF LYSOSOMAL ENZYME EXCRETION IN IIUMAN URINE Whether or not the high levels of urinary jlucuronidase observed in blad- der cancer patients are a cause or a consequence of the disease could cooceiv- abiy be resolved by determining whether individual variation in the excretion of this enzyme is genetically determined. It ao, whether bladder cancer patients have a jienetic predisposition toward bigh enzyme excretion could be estab- lished by measuring enzyme excretion in their progeny. New aasay procedures utilizing fluorometrk substrales were Ihereforn developed, in order to measure A-glucuronidaae. S-tialactosidase, sip-Salaelaidase and A-helosaminidase eacre- tion. 77rc assay conditions for a-glucurooldafe eliminate the previously trouble- some problem of interference by both low and high molecular weiilht inhibi- tors. The urinary kvels of the four lyaoaornal enzymes vary coordinately, that is, although their absolute levels (unha/mg crestinine) vary during the day, and from one day to the nest, the ratio of one enzyme to the other remains relatively constant. The lack of oornlalion between their plauna and urioe levels and their high enokeular wdghl, both suggest that thew urinary encyrnn do not derive trom gkMeerular fUtretbn. That then is also no eoord)nacy with lactate dehydrotenase, nsoreover, suggests that they do aot corne from eafoli- aled cells. 11 is proposed that they stem from lysoaornes extruded by epithelial cells into the lumen of the proabnal lubuk. Total enzyme excretion abo varies considerably, as shown in a population of 125 healthy adults. However, both total enzyme escretion and coordinacy ratios follow logarithmic distribution, suggesting that lhey are the resultants of many tacton, each having a relative or proportional effecl on enzyme eacretion. ralten. K. and Petenon, 1. The /owrna1 o/ Clinical I nvesrigarion 61 :711-762, 1978. OOther support: National Institutes of Health. From the Departmcnt of Mokcular Riobgy, Roawell Park Memorial Institute, SuRab. 11. T/re Respiratory System DEFENSES OF THE LUNG: A DYNAMIC OVERVIEW . Lung defenses include a number of inlerrelated nonspeciflc ewd specific systems, from the nose and mouth to Ihe alveolar-eapi0ary parenchymal Inter- face. '1'hese are intended to isolate or eachrde noaioua inhalanta from the deli- cate mucosal and/or epithelial cellular membrann, either throulh mechankal barriers (e.g., mucus) or by physical removal mechanisms (e.l., sneezing, coughing. reflett bronchospasm, mucociliary escalalor, or macrophage removals). Within the lung, substances can be detoxifkd locally by interacting with anti- microbial protein secretiorn, with airway and/or vascular enzymatic mech- anisms, or with the complex airway and parenchymal immune defense systems. lhii e.temive review empha+irrs the importance of interactions between ciliary (uncnon sr>,f mu.rn anJ between mscrophsges and other lung cells. llre role I of the various mucus constituents in airway defense responaea, however, re- mains to be fully clari(kd. It is apparent that pulmonary alveolar macrophaolu may have the potential for many functions in addition to that of primary alveolar defense against microorganisms. They may also have a key role in the pathogenesis of various inflammatory destructive and fibrotk lung dreasea. Furthermore, it is recognized that while mucus-producing cella and plujocyles are crucial to the defense of the lung, they may also harm the host. Broader comprehension of these integrated defenses (and potentially injurious macb- anisma) should further clarify the pathogeneais of lung diaeaae and lead to improved treatment. Croas, C. E., Kaiw, T. and L..t,1.A. In: C1ark, M. A. (ed.): lrln.owory Dtreasr.: DtJenu MecAan(snv and ropa- larlonr at R64. Tobacco and NedrA RneareA InrNrrtr Sympos/re.-2, l edng- 1on, Kenlucky: University of Kentucky Printing Serviees, 1979, pp. 14-59. Other wpportr Primata Center Ilw Grant eed Ewvironmental Protectio. Agency. From the University of California Sclsod of Medicine and the California Pri- mate Research Center, Davis. LUNG HOST DEFENSES: A STATUS REPORT The lung has a complex and ealensive system of defenses that work. tirelessly to purify inspired air and keep pulmotaary (iaaue free of iefectio.. Since the major components of this system have been identified and well de- scribed, present research is focusing om the eoardination of these parta or on the integrated function of the whole syatem, particularly on the cellular iater- acliorn, generation of local inMnune «spoesea, and nnpUfyinR-inhibitory faa ton regulatinR inRammatiow. A tab/e of httrR host defenses b airway challenge presented here lists 11 surveillance mechanisna and two augmenting mecha- aisms,-iniliation of 6emune responses (humoral antibody and cellular) and generation of an inflammatory response (infiu: of polynarphorysckar Rraw- locytes, eosinophils, aod 7 lymphocytes). Colkaivey, the ekments of this de-ense ayslem work smoothly and continuously. Increaaiagly, however, ea- amples of isolated immune deficiencies are being found which are associated with recurrent pulmonary infections. Complement factor deficiencies (C,), abnormally functioning cilia (Kartagener's ayndrome), absence of immuno- globulin (IgA), and abnormal kukozytea, all seem to predispose to lung i.- fections. These sekctive deficiencies aetve as splendid probes for asseaaing tl.e Importance of individud components in lung defense. However, the fact that lung function can be maintained in the face of one or more of these dcbckn- cies serves to dramatize the overall ydaptability of the lung defense system. Reynolds. H. Y. C/irst 73(2):219-242, 1979. Other support: National Institutes of Health. From the Pulmonary Section. Ikpastment of Medicine, Yak University School of Medicine, New Haven, ('onn. 23 22

VASOACTIVE SUBSTANCES AND THE LUNGS: MOLECULAR MECHANISMS There is uo question that gas esch•nhe ir the lungs' primary function and that interference with this process is the m•ior determinant of mortality in the microembolism syndromes. A variety of other metabolic reactions conducted by the lung., however, may well contribute to the clinical eapression of sy- dromes and may play a role in the development of pulmonary and systemic vascular drsfurxtion. For esample, lung endothelium has a signi6cant role in the regulation of circulatory contx•IrNio•w of kinins, angiotensins, prosta- =landins, and adenine nuckolides. Other mechanisms are still unckar, however. 7 he importance of pulmonary endolbelirw in the inhibition of microthrornbi formation and the role of other cell types i• such activities remain unknown. Still other questions that are posed an: ts the aMi-•ssretNing aSen1, pros- lacyclin, made by the endothelial ce1Mt Ca• these celh inactivate the pro- aggregatory faclor, ADP? What is /he importance of pulmonary angiolerain converting enzyme In the development of renal dysfunction in the delayed microemholism syndrome? Would know• inhibilors of this enzyme alkviale or eliminate renal InsufRcieacyT Ecaminatioo of what happens 7o the "oon- ventdatory" functions of the lung during the development and course of the early microcmbolism or delayed microernbolirm syndrome could be informa- li.e. It may help define the humor•1 meehanisms through which microemhol- ized lunp affect renal function •nd, perhaps, slso the processes through which permeability factors are teneratcd. Ryao, 1. W. and Rr.n, U. S. In: S.Ideen, T. (ed): The MkrormAdinn Srndro.ne, Stockholm: Aheqvist A Wikull /nlernalional, 1979, pp. 213-221. Other sMppsrtr U. S. Public Health Service and the Hartford Foundatioa. From the Department of Medicine. University of Miami School of Medicine. Mismi, Fl•. VASOACTIVE SUBSTANCES AND THE LUNGS: CELLULAR MECHANISMS This review summarizes what is known about the nonventilatory functions of the lung. UUrntructursl studies demonstrate various specialized structures and microenvironments suitable for the processing of vasoactive substances. While many of the known "para endocrine" functions of the lunp can be attributed to the activities of the endotheli•1 celts, their functional limit in the processing mechanism of Mrrnwnes, prohormones and other escitatory sub- stsnces, or in the synthesis of vasoactive compounds is unknown. l7rcre are, moreover, pulmonary subslances such as thrombosants, which are released by the intact lung but are not produced in large quantities by endothctial cells. Consequently. it ir eksr that more remains to be learned abont !.pecifk mela- 1Jhc at iwaws 4•f puln.un.ry enduthebsl cells and even mure alrwl such ae- 1t.sly in od.rr pulm..n.# y ..uulu tell typcs a Rr.n, U. S. and Rran,1. W. In: Saldcen, T. (cd.): The Mknoenrbl4n. Sywdrowre. Stockholm: Akaq.lrl & Wiksell Intern.tional, 1979, pp. 22)-272. Other support: U. S. Public Health Service and the Hatt(ord Fouedatio•, From the Department of Medici•e. University of Miami School of Medici•e. Miaml, Fta. SYNTHESIS OF PROSTAOLANDINS BY PULMONARY ENDOil1EL1AL CELLS Although it ha. beew known that lunp rekaae relatively IarRe quantities of prostaplandin-rclatcd wbstantea following embo*atioa or induction of u.- phrl•sis, nothing ia known of the oellalar eilea of spethesis of the .ubMa•cea released by the lunps and nothing in knows of bw tA• prwt•Rl.ndin-relatcd substaaces gain access to the vascular space. 1s this attempt to deterei•o whether pulmonary e•dotheli•1 eells contribute b 1!r eRlut of proet•pla•di•- «lated subetanees, cultured bovine pulmonary arterry esdothelial eells wen i•- cubaled for two houn with 1-r'C-ar•chidonk acid in mediurn 199. A bkr•k, containing rrC-•r•chidoaic acid i• medium withoat «lle, was dso incubated. The diiference chromatoprnn obtained by eublracyinR the blank c.p.m. /ram those of !he esperiarcnt•1 showed at least .it s4C-lsbekd emtaboliw, but .• peak corresponding to ar•chidowic acid. PbE,, • major tneuboWe, /ornnA at a r•w of 0.33 as" of cell protei•/2 hows. POS, showed up m• second metabolite. Having established th•t pulmonary artery endotbelial celk+ i• cu{- ture are capable of inet•bolisini aradddonie acid to yield POE and related wbstaeces„ a search was slarted for sikU of We fatty acid eyc1o-osy8eaase. Among attempts reade lo 1•calize nAotfsdi•1 eyclt..oargenaae using combi•ed ttarphok>tk and biochemical IecMiques, both ekctro• microscopy and nrto- radiopraphic audies wer• do•e. The results of ther studia ahow that bovi•e pulmonary artery endotbe4al eells are e•p•ble of forming proet.pla•di•a ana related substances Irom 1-r4C-arachidoe•1e Ibr•ugh the action of •n e•spme system situated on the endoplasmic retie.hw.. P(1Es i. a major metabolite, and a substance which cochronutographo with R-keto-POFr., a breakdow• product of PUI,, is dro formed in highly variable qw•tities. While these ra- wlts indicate that pulmonary endothelial eeqk c•o account for p.rt of tbo etllua of prostaglandin-like substances from eabotiud or ao•phylactk lunp, they provide no information on the cellular origins of thrombo:ane A,, likely to be a primary medi•tor of •n•phylaui., or of other subuanea which may occur in lung venous eAlue•u bt much higher eoneentraliona than those of POEr. ~ Ryan, l. W., Rysn, U. S., llablisto•, D. and Martiq L. Trauarrbns of rhe Assorl.ulon o/ Arnrrk.w pApskbnr aei:74)-730, 1978. PtbeT support: U. S. Public Health Service and the Hartford Foundation. From the Ikparrment of Medicine. University of Miami School of Mediciae, Miami, Fla. :4 25

ISOLATION AND CULTURE OF ENDOTHELIAL CELLS FROM THE LUNOS OF SMALL ANIMALS Th. object of this study was b obain pulmonary esdotheliar eet6 from small a.imale commonly used 1o stud7 the metabolic tales of ho -rrwnes and other eacitatory subrtances, and to fLd a iray of coNectin6 thesi cdts from th. pulmonary vasculature. Teeh.auq rred for isolNing and culturing endo- thelial cells from the lungs of rabbitR fltrfnea piP and rats aru deacribed. Retrograde perfusiom of blooddra hrys with buffered aaline .olution cootab- In6 colla2enase was used to eolket t!e oeflr, which were then char rcterized by light microscopy, electron mictoeoopy of tldn sections ud wrfrce replicas, and by the presence of antiotesiteo.verting enzyme (ACE). The ACE was assayed with 'H-bensoyl-Phe-Ala•Pro sa We wbstrNe, and IocalireG by Indirect immunnAuorescence, using guinea p4 qdolbelial te1N incubated with rabbit .ntlbodin to 6uinca pi6 hm6 ACE followed by 6oat anti-rabbit globulins con- jutated to Auorescein. The Iechniqw described here allows the use of small animals for convenience. It can also be used os larger animala, however, and even humans from whom pulnwwary l,ndothadum may ona day be obtainable by open biopsy. Habli.tow, D. L., Whitaker, C., Hart. M. A., Ryaw, U. S., and Ryan. 1. W. AnKrkan Reritw o/ Rtrpiratory Disease 119(6):e3)•i6~, I979. Oth.r wrportr U. S. Public Health Service aed the Hanford Foundation. From the D.partment of Medicin.. University of Miami School of Medicine, Miami, Fla. AGE-RELATED CHANGES IN ELASTIC FItiERS AND ELASTIN OF LUNG Numerous physiologic studies of Ilumm lungs show decreased elasticity with aging, but the morphologic and chemical bases of these changes In both the elastic Iiwm .nd the orpnization of the elastic tiber network are not clear. The effects of age on the hrnp of .n inbred strain of mice (tiA1.t!/e) were studied (I) to determine the .Iterations in the morphometrically estab- lished total elastic Aber length (TFL) and the chemically measured elastin content in nonemphysemalous aging hmp; and (2) to correlate TFL and el.stin content with static compliance of excised lunp as a function of aging. According to the resuhs: (1) static compliance Increased with age; (2) TFL increased with lung expansion in ap- and sea+natched mice, indicating an aaial extension of elastic fibers (but in aging lungs fiaed at a distending pres- sure of I S cm 11 ,0, l FL did not change significantly in spite of an age•related increase In lung volume); and (3) both lung collagen and elastin content de- creased with age, she tatter indicating a loss of elastic Nben. The virtual absence of p.eudoelastin fihers in the lungs of aging mice, as determined by histochemical technipoes, may account for the differences in elastin content of aging human and rnmne lungs. Additional studies of the pseuduelastin eon- lent oi human and animal lungs are indicated. Ranga, V., Ki.onrrmnn, !., Ip, M. P. C., and Sorensen, ). I American Review ol Rrsoiratory D/sraa 11i(3):)69-176, 1979. OtAer.rrprtr Natlon.11m1itute of Ea.iro.mestal Health Sciences. From the Division of Pathology Researeh, Dep.rtmente of Patbolop. St. Luke's Hospital and Case Western Raerve Universily, Cleveland. EFFECT OP TOBACCO SMOKE ON THE METASOLISM OF RAT LUNG The composition and metabolic activity of rat hteti parenchyma Jter 30 days of in vivo espowrre so cigarette unoke was i.Malipled. University of Kentucky research cigarettes (brand IAI, 25 mR tar, 0.3 mR aiootisro; ..d brand 1 R 1, 30 m6 tar and 2.2 mR .iootia.; wei6ht eapte.red per cigarette) in a Wahon smoking machMe wsre used for one rnontlr ow two scheduhy: one cigarette twice daily or three eipretles hvke daihr (7 ptiQs/dprette). Oan group of aeirnals served s Intaet eootroY, while another control group was introduced Into the machine but aot exposed to Mrwk.. AR uimalr wa. wel6ht-matched at the beginning of tM eaptximed and had free aocets to food and water. At the end of th. 30 days„ Ipe weiRA1 pis was noted i. the machine control rats (2!%). th. IAlpposi0 group (I0l6 ), and th. IRI- eapoaed rata (26-10% ) tha in the intact eoatrok. Study of the eAeet of cigarette smoke on the uptake amd metabolism of sH-pahnital., a11-kudee, Ur'C-glucose and r'C-tlucoe.min. I. hmg slioa showed that .either p.ImNate uptake .or phoaplwlipW aynthesis was dke1N4 E+tposure to IAI eipr.tlse increased sCO kvds by 25% ard Mpid leoorporatioe of glucose by 30%. The IRI brand had no eRect on Rlucosm metaboliarr, but lowered prolsi" nrnth.sis by 30%. Both braaAr, horrev.r, tatreA a two-toW ixrss. In glyco- protein synthesie, which wr probably rdaled bo hi6her productio: of nwcus after exposure to amoke. H.w»osA, AI., Shechier, Y. and Hamosh, P. ArcAires o/ Ewvlrewiwrearat HedtA )1(1) :17-27, 1979. From the Department of Physiolm and fliopAya{o. Georgetown University Schools of Medicine and Deetistry, WaahinRlottr D.C. NEUTAOPHII. LYSOSOMAL ELASTASE ACTIVITY IN NORMAL SUt!)ECTS ANI) IN PATIENTS WITH CHRONIC OOSTRUCTIIVE PULMONARY DISEASE The degree of circulating posymorplwmrckar, h:ukocyte lywaornd elaalw activity was determined in asymptomatic aduUe with normal lung funtlitrok is patients with chronic obstructive lung disease (COPD) and in alphao-an8- trypain-0dlcient individuals with and without evidenee of COPD. The resulting data were analyzed for any relationship between the presence and severity of COPD nd newrophil lysosomal elntase t?onoedratqra. In addition, the eo- zyme's activity was repeatedly determined at 1 to 12 month Intervals i, a population of normal subjecls and in a group of COPD p.tienta. No diller- ences (n neutrophil lysosomal elastase activity between smokers and nonsmoken 26 27*

were noted ie either normal subjects or patients. Siknifkanlly greater degrees of enryme adivity, however, were found in patients with Pi M phenotype and COPD, suggesting that circulating dastaae activity may be involved in the mechanism that damages lung elastin and, thus, ie the pathogenesis of pul- nae.ry emphfsenu. Rodrigues:, 1. R., Sesh, 1. B., Radio. A.. Lie, 1. S., Mandl, 1., and Tr.Jwo, G. U. Anwrk.w Review of Respiratory D/arosr 119(7) :409-417, 1979. Othor w".rtr U. S. Public HeaM1 Senioe and the Stony Wold-Herbert Fund. From the Departments of Medkir and Obstetrics and Gynecololy. Columbia Univenity College of Physici.r i Surkeorrs, New York. ELASTOLYTIC ACTIVITY OF ALVEOLAR MACROPHAGES IN NORMAL DOGS AND HUMAN SUBJECTS Uneertainty remains as to the precLo kver and variability of alveolar mscrophade elutase activity aemng earoal stobjectsboth smokers sed rws- smoken. Is addition, the capacity of hunus alveolar macrophakes to grow aod divide /n vitro has never been defined. This study measures cleanse acliv- ity in freshly Iavated human and dos alveolar macrophages from normal sub- (ecu, in order to determine its magnitude ssd variability. There seemed to be no relationship between a history of smoking and cleanse activity. /n rlrro, the macrophagcs released measurable and similar levels of elastase into the culture medwm over a period of three months. The cells underwent mitosis and demonstrated phakocylosis. Synthetic sile-speci0c elastase inhibitors (chlora methylketones) btally suppressed elastaae activity. According to these dats, both canine and human alveolar macrophates from normal subjects show ape- cilk elastase activity regardless of the smoking history. This activity appears to be generated continuously by the maerophages themselves rather than in- corporated from external eouneL Green, M. R., Lin, J. S.. 8erma., L. it., Oanan, M. M., Cerrets, 1. M., Maodl, 1., and Torlwo. G. M. Jownd oJ L.Aorerory.nl Clinical 6felk4w 94(4):31l-362, 1979. Other anrprts U. S. Public Health Service and the Stony Wold-Herbert Fund. From the Departments of Medicine and ObNetrics and Gynecology. Columbia University College of Physicians A Surgeons. New York. I UNO INJURY INDUCED BY LEUKOCYTIC PROTEASFS Human polymorphonuckar neutrophilic kukocytes (PMNs) contain lar9e amounts of neutral proleases. Inslillation of one of these. elasuse, in the purJ'ied form. rnto dog lunts ro rivo prodraes degradation of elastic fibers and orher atvcolar scpral cwnponents and leads to anatomic changes similar to I those found In human pulmonary emphyepna. Cigarette smoking may 1e1er- (ere with the regulation of PMN elastae activity by alveolar antiprotesses. This is supported by the observation that the oxidizing activity of tobeooo sreoke inactivates .r-proldnne iwhibilor M drro. Low levels o( elaaolptic proleases are also secreted by snaerophalles. Cultured mouse sn.crophatia aa- posed so cigarette unoke for a short period of tMse demonstrate an hxreased rate of cleanse accretion. This enzyme is not Inhibited by .,-proteinass W hibitor or by .; macroff{ob.alin. This unusual property of wmerophage ehstase may facilitate the breakdown of elastht o.er a prolonged psriod of time is spite of very low levels of enzyme. A unified hypothesis o( Me role of cleanse in the patholenesis of puhvwn.ry emphysema is proposed whereby two diQer- ent puhways for the production of hmd injury by cipeNts amoke ara envi- sioned. One of these is mediated by PMN dastase and the other by aucrophage elasta+.. Future research should resolve which pathway ia the key determinant of lung injury in the smoker snd how it eam be blocked. J.nol. A. er ed. American Journal o/ tuAoWq 97:111-I16, 1979. ~ Other ar".rtt National Heart, LunR and Blood lartitule. From the Department of Patholop, State University of New York d Stony Brook, Stony Brook. CIGARETTE SMOKE INHALATION DECREASES a,-ANTiTRYPSIN ACTIVITY IN RAT LUNG , Aqueous extracts of cigarette snake ineqivale human ..entitrypsim b rirro, suggesting a relationship belween eiprctte smoking and pulmosry emphysema. lhe experiments reported here were intended lo determine whether cigarette smoke inhalation aho inaetiratea lung ar-antitrypsin in vivo. Results show that brief inhalation exposure (thres to sia puQs of cigarette smoke) of rats significantly decreases the cleanse inhibitory capacity (EIC) per milligram of .,-anlitrypsin In lung lavage fluid. This loss in EIC could be reversed by a reducing .teat, suggesting that osidation of the Inhibitor in the lung by some cigarette smoke component mi6ht be the responsible mechanism. That o:one-tolerant animab .re not a0eeled by cigarette smoke supports this hypothesis. Human serum obtained inunediately after smoking also shows a decrease in FIC. It may be that transient imbalance between lung proteasee and their inhibitors caused by smoking injures the alveolar walls and that repeated imbalsnces of this type slowly deform the alveoli, eventually leading to disease in wn.e chronic smokers. JanoO. A. er d. Science 206:1111-1114,1979. Other e.r'prts U. S. Public Health Service and the [kpartment of F.nerar. From 1he Ikpartment of Patholonr, State University of New York at Stony Brook. Stony ttrook; and the Medical Depsrtment, Brookhaven National tabo- ratory, Upton. N. Y. 28 29

STUDIES ON ELASTASE SECRETION BY CULTURED MURINE MACROPHAGES !n the shdies reported here, mouse peritoe,eal exudative macrophages (PEM), cultured ie the presence of aqueoru dgarette smoke e:tracts, increased their production of elastase in a way that was dose and time dependent and could be inhibited by cycloheaimide. On the other hand, untreatrd mouse alveolar macrophsges secreted elastase at a rase equivalent to that shown by smoke-treated PEM and could not be further stimulated by ,hort-Ierm exposure to aqueous smoke extracts. Even psaler baseline elaslase .ecre/ion vras sbown by exudative alveolar mscrophallas oblained from mice previously -raccinated with complete Freund's adjuvaet. Wben the susceptibility of macrophage elas- tase 1o inhibition by endogenoau aMiproteases was tesled, h was seen that macrophage elastne was only poorly inhibited by a,Pi (human) : though it was definitely inhibited by both PiMM and PIZ sera. Is other work, the ef- lects of purified a,M on the eladolytic activity of mare macrophnle condi- tioned medium were examined. The rewib clearly denwnstrale 11at under , conditions suflicient to inhibit porcine Paecreatie elWase, the m.erophaje enzyme was not Inhibited by a,M frorw either hunun or mouse ma:rophaRes. Although these results do not explain the Iwhibition of m.crophage clauase by PrZ serum, they do show that macrophase elsuse ia tewre resist,mt to 04 major endogenous inhibilors than are other .eulyd protenes of lAalocytie cells. This suggests /hat, in dgarette smoten, 1be macrophage en:;ime could play an important role in lung injury. Whi1e, R., White, 1. and /ano#, A. INSERM t4:1S9-16S, 1979. Other a.rr.rrr National Heart. Lung and Blood Institute. From the Depertment of Patholop, Sute University of New York at Stony Brook. Stony Brook. ALBUMIN MICROSPHERES AS CARRIER OF AN INHIBITOR OF LEUKOCYTE ELASTASE: POTENTIAL THERAPEUTIC AGENT FOR EMPHYSEMA Succinoyt-Ala-Ala-Pro-Va1CH,C1 has been showa to be one of the more potent inhibilors of human kukoc7te elatae, an enzyme that has been im- plicated i. king tissue injury leading lo emphysema. While this inhibitor seeau to cAer promise as a Iherapewie apn1, (ts eAecdvenea would probably be greatly enhanced il it could be targeted directly to tM /unRs. In this paper, the sequence of reactions involved i. the covalent .ttachmtnt of the inhibitor to human albumin mkrotpheres with extended side ehains Is described In detail. The insertion of side arnw of various lengths showed that ma.imum enzyme Inhibition was obtained when the paar arm was al least 24.7 A in length App.o.imetety, 10 mokcules of the inhibitor could be attached to each mulecule of ./bum.n, wah derrvalured rnierospheres were cap.bk of InhMb!ting .ppruamatcly one mok of els.r.ae per mok of .Ibumin, which n comparable w rhe inhiblory .crrvrry uf ., antitryprrn Etperoments in vivo in which rats I were in~ected inlravenoudy with radiolabeled mkroipheres to which 16e I.- bibitor h.d been attached showed a rapid and eaclusive uptake by the luop. About 40-50% of the injected microspheres subsequently remained in 1ba lungs with a hdf-life of approximately 17 days. Although these studies wera specifkally designed to asseas the feasibility of altRhinR an elastue Inhibitor to a carrier that could be targeted so the lunp, the broeder implicalions of this approach deserve careful consideration. Martodam, R. R.. Twunasi, D. Y., Gcncr, /. E., Powen, J. C.. Nisbino, N., and Krejcaret, G. • Proceedings o/ the National Academy of Sckncer o/ the United St.as o/ Amerka 76(S):2128-21I2, 1979. Other at pr.rt r NationallnuNutes of Health. From the Department of 6iochewriqry. College of Biological Scienoes. Unt- versily of Minnesoia, SI. Paul; Department of Cheraislry, (korga Institute of Technolop. Atlanta; and the Nuclear Medical LaborNory, 3M Company. St. Paul. I QUANTITATIVE CHARACTERISTICS OF THE FEYRTER CELLS At ID NEUROEPriHEL1AL BODIES OF THE FETAL RABBIT LUNG IN NORMOXIA AND SHORT TERM CHRONIC HYPOXIA This report attempts to estimate the number of single Ferrter (APUD) cells ud neuroepitheH.l bodies (NEes), or grouped Feyrter eelly is the lungs of 26-, 27.5- and 29dars feluses dueMg aotrnosia and short-term chronic hypoxia. The possibility that these cell numbers vary in fetuses from short- term chonically hypoxic mothers was also i.vestisaKd. The nast sisnilkan/ variation was found between the 26- and 29-dar fe/use., the marked decline in the apparent number of lhese «Ms suggesting a response to increased bypoa- ernia. The lunp of fetuses from shor/-1erm, ehronicaly hypoxic nathen, fur- thermore, contained fewer of lhess eells thaw those of feluses from nornrozie mothers. These observations apee with the resuUs of previous studks in short- lerm. chronkally hypoaie neonN#1 rabbils, and suggest that induclion of br- poaia in the mother increares hypo.enua in the fetus, thus stimulating tbe secretory activity of Feyrter cells ud NEM. This in turn eupporu the hypotb- esis that the Feyrter cells and the NESs may play a role in the mechanism of hypo.emic pulmonary vasoconsltiction in Ihe fetus. In addition, the oMerva- tinm on letuses of normoak rabbib provide base line data on the chroeoio{kal imporlance of these cells. Hernandet-Va.wret, A., WIN, I. A.,and Quay, W. d. Celt and T/rrrc ['uhrre 159:179-1 t6, 1972. Oflr.r .u'rortr University of Wisconsin College of Agricultural and U/e Sciences. 1'rom the Dep.rrment of Velerinary Science and Nnuoerwlocrine Section. War- man Center on Menlal Retardation and Human laevcloprnent. University of Wiseomin. Madieon. 30 31

HUMIDITY AND THE ANESi71ET1ZED PATIENT This study attempts b corrslaN the eybmorpbdogic chan=es caused by a)ack of moisture Ia anesthetic pw with the incidence and severity of poN- opcrative complications. A point soocin8 system wr tned to aaseas theae changes in 202 patients who breathed either dry or humidified psa for a period of 223 t 71 mia. Results abow that pouoperative pulmonary complka. tiorr increase .bng with damage 1o Ib cBiated epitbelitun. 71w incidence of theas cornplicatbns was reduced, htwe.er, a Ibo humidity of ehe administered gases rose from 0 to 32.5 me H,O/L Ro1h btwt low aed the incidence of poM- operalive shivering wer. aho higher r 1V moisture conlenl of the anesthetic dropped. It appears lhat the u.s of dry a.albetk pnes for operations laaing longer thaa one bour may be h.nafui. The Weal level of humidification aoema 1o be that which returns to tha ptket tM e:act amount of watcr lost during eapiration (.u per saturation humidity M 72'C). Ttiis has three arhanlaAes: it (1) decreaea damage to the tracheal nwoaea; (2) lowers the pottoperative eomplicaliona rate; ud (3) vlrtu.lly e0rwi..lea poatanesdretk shiverinA (thia ia not desirabk, howevrr, in the presesoe e/ tubsisaat hyperpyrsxia, whea beat loss should be increased by all possible eeaes). Some improvemeatb In the design of anesthesia appuatus that would 1.ke the control of humidity output into account are suggested. CAalon, !. et d. Anrit/ierldopy 30:193-195, 1979. From the Department of Anatbealology, New York University Medical Cen- ter, New York. . 111. He.rt and Clrer/.tiow EFFECT OP DILTIAZEM. A CALCIUM ANTAOONIST, ON MYOCARDIAL ISCHEMIA Diltitaem hydroehloride. a potent eakiwn aotaBonist, was a+.ses+cd for its elRcacy ie reducing the effects of myocardW Ychemia. Melabol c/wctioe wilhie both behemk auid aoalachew+k titrue was analyzed a.d compared iN two groups of dop: Orvup I was sacriboed atkr 60 minuks cd regional bchemia without receiving tay of the dru8; Oroup 11 wn given diltiaus after t0 mi.utes of iachemia a.d sacriboed 30 ati.utes later. DiNiveat ditniM- Wred the injurious effects of i.cbetnia In several ways. it (1) reduced the de- ereaw is adenosina-S'-Iriphophate by baH; (2) decreased the Inaibition of anaerobic Alycolysb; (3) lowered 1inue lactic and free Iatty acids kvela; and (4) Improved the eontractitity of Alyoerinated heart muscle fiben. MNo- chondrial (unction, however, was wt dleeled. Mitodwndrial oaygen uptake, respiratory control Red eakium ion bindio8 were equatly reduced in both groups. It Ia neverthekas clear that even if iu benefkid action does not ealend to all the pathways of myocardial t»etaboliarn, dihia=em bas the capacity to minimize the adverse effects of acute ischemia. Wnshasr, R, Ashikawa, K and siny, R. J. The AnKrkan lorrnd o/ Cardioloty 47:1177-1143, 1979. 0t6er au'porrt Tbe Hoover Fouadatioa From the Huntington Institute of Applied Medical Research aod the Califor.ia Institute of Technology. Pasadeoa; and the University of Southern Califor>tla, Laa Angeles. INHIBITION OF CHOLESTEROL UPTAKE BY THE ARTERIAL WALL IN THE INTACT ANIMAL 1t has previously been demoastr.led that 7-ketocbolaleroi, a t.oi-physb- bgic cholesterol aaabg, Inhibits the /n r/tro uptake of cholesterol by perfused human tad animal arteries. This report pre.eets evidence indicating Ihal 7- ketocholesterol effectively inhibits cholesterol uptake by the arterial wall In vivo u well. Isolopaa blood preincubaled with the sterol was infused Into in/act rabbila along with some of the anLwab' own plasma codainhg radio- active cholesterol used as a lraoer to measure arterial uptake during two successive Infusions. Thu., each rabbit served in i1s own eo.tral. le the presence of 7-ketoclak.lerol, the mean cholesterol uptake by 1he carotid and femoral arteriet was 11 t 3 amoln/ R tiswe an oppored to 3) t 3 enroks/R amue without it. Tbe tnean blood 7-kelocholalarol oonce.tratiaa ranAed from 34 to 87 aepks/ml. Preliminary In wbro esperitneats abowed that preiecu- bation of plasma -with particulate 7-kelochokslerol also inhibited cholesterol uptake by perfuaed pig eoroo.ry arteries eves whes /his particular fraction was removed before perfu.ioa. Tha 7-kdoelakslerd bindo to the plasma lipa proteins and bas an evee hiAhec aAl.ity, for red blood cells. dlnp. R. I.. Sarma,l. S. M. n.d Chn, 5.1. I Arrery 3(/) :14-28, 1979. ~ ~ OtRer wrrertr The Hoover Foundation. From the Huntington Institute of Applied Medical Research and Huntington Memorial Ho.pital, Pasadena; the Division of Chemistry and Chemical Er Bineering, California Institute of Techaology, Pasadena; ..d tAe University of So ybern California, Los Angeles. SERUM CHOLESTEROL ESTERIFICATION IN HYPERTHYROIDISM AND HYPOTHYROIDISM Changes In the level ted compoailios of plasma lipoproteir h.ve bee noted in cases of thyroid dysfunction. This study atteppts to determita tha effects of altered lipoproteiw metabolism found iM thyroid disease on kcilhYt:- eholederd acyllransferase (I.CAT) activity. T/r rate of serum ehokNeroi esterifkalion was measured i. 20 4scallhy bumaw wbjeels withou/ a fanJly history of lipid or thyroid disorden and used as caMrd data. When thesa were compared to values obtained in 17 hyperthyroid and 10 hypothyroid h- dividuals, there were no signi/kant diRerenbes in the cholesterol esteriAcaliow rates, but highly signifkant (p<0.001) differences were found in the fractional tales. 7hese were higher than normal in the hyperthyroid group and lower than .ormal in thc hypothyroids. TAerapy did not seem to have any consistent eRects on the cholesterol eslerification rate. Thia either increased or decreased 32 33 •

I as patients became cuthyroid, reprdkss of their original status (i.e., hypo- or hyperthyroid). The fractional esterifkatioe rates, however, did show a clear trend during therapy, always increasing or decreasing in hypothyroid and hypenhyroid patients, respectively. Lacko. A. O., Muks, A. D., Ruteeber6. H. L„ and So/o0, L. A. Hormone ewd hlet.bolk ResercA 10:117-131, 1976. OIAa ur'prtt Administration o. Ati.B. From the Department of Mediciee, Tem#l. U.iversity Health Scknoe. Ceeter, Philadelphia; and Tesu College of Osuopahie Medkiee, North Te:as State Uaivenity, Deebe. INTEGRATED CAROTID CHEMORECePTOR AND PULMONARY INFLATION REFLEX CONTROL OF PERIPHERAL VASOACTIVITY IN CONSCIOUS DOGS With 14 mongrel dogs in the corrcious s1.1e r experimental subjects, the Iateraction of carotid chenwreceptor a.d pulmonary laeatioa reAe= eootrol of vascular mpn.nes is the aseaeateric, resul, and iliac beds was examined by eomp.ring respooses to chemoreceptor stimulatio. (ielracarotid ia' ioo o( nicotine or cyanide) during spontaneous and controlled respiration. ~s were dso studied in the eoescioru state after (1) bets receptor blockade with pro- prarwlol, (2) cholioergic blockade with atropiee. (3) bistaminergic biockade with tripekmamiee, and (4) alpha receptor bbckada with pbeetolamise. Also, to e:areine the effects of c'henareceptor atinulatioo in the absence of changes in ven/ilnios and secondary refk>< effects of pulmonary infiatioe, 11 dogs were sludied during succinykholine infuiion with controlled ventilation. Results showed that rHrKarotid administration of aieotiee or cyanide in the intact conscious dog elicited a biphasie cardiovascular rsspoese. The Mt phase was charaderited by bradycardia, as iecre,ae in re jaaal resisla.on, and ae in- erease in depth and rate of respiration. The seeord phase followed the rn- prralory changes aad was characterized by tacbyeardia and a decrease in regional resistaaoes. Alpha-blockade with pheBldami.a nearly abolished the early vasocosstrictios and 1.1er vasodilalio.. When tha ehanoreeeptor-iodured increw in respiration was prevented. or after bilal.ral va6olomy. there vru significantly more vasocomtriction in all three beds eed no later vasodeatios. lUere(ore, io the coescious Baimd, carotid cheworeceplor stimutuio. resups ie a bipbasic vascular respos.e, while the pulmonary in0ation re/kses are sufM ciestly powerful to atteouate the initial v.soooastricbr resporne and to reverse the coeulridion to a later period of ieteese vasodilatioe. Rutherford. 1. D. and Yuwrr, S. F. Circulation ReuarcA Il ( 2):20Q201. 197t. Other arrprt 10. S. Public Health Service. From the Department of Medicine. Harvard Medical School and Peter B:et Brigham l/ospNal; the Ikpartment of t:ardioloty, Chitdres's Hospital Medi:al ('enter, Boston; and New Nngland Regeonal Primate Research C'enter, Southboro, Mars. 14 CONTROI. OF THE MYOCARDIAL CONTRACTILE STATE BY CAROTID CHEMO- AND BARORECEPTOR AND PULMONARY INFLATION REFLEXES IN CONSCIOUS DOGS This study examines the extent to which the carotid c6ernoteoeplor reas= re6ulates myocardial cootractiNty in conscious doa aad, Nlempta b Ideaihr the eateat to which these effects would be modiAed by Ibe ooeooedtaM b- ' creaae in respiration aod ~tinwlalio. o( pulmonary idlatioa aRere.b. Catheten were Implanted in the left atrium and eorta of 19 dogs, and ths eaperiwteefla were conducted three weeks to two monthe postoperNively, whew ebe do6s were again vigorous and healthy. Cber.oreoep/or atimulation wae aecooplia6ed by lejectioe of nicotine (0.2 pg/kg) or sodium qaride (2.0 r3/kt) irto the (.lracarotid catheter. Theaa two qenla used for Miewlatie6 1he carotid esemo- roeepbr relks isduced similar e/fena e. heart rNe, LV ..d arterial preaauna and dr/fr. B.roreceptor un{oadieR was .ccornpli.hed by ieBati.6 the hydrau- lic occhrders implanted on thm eonreos carotid arterirs. Dogs wese aYo adsdied in the co.scious state after: (a) cloli.er& blockade with atropiee; (b) beta adrenergic receptor blockade with proprawolol; and (c) eorobined eholimrgie and beta adrenerpc blockades. The adequacy of beta receptor bbekade wr tested with i.oprotoreool and that of cholieeritie blockade wr te.ted with a¢tytcfaiine. In .urnmary, rauNs showed that the carotid ckKmoreceptor re- Bez in conscious dop, when ainwlated, elicits a si6nibcaet increre I. ruyoe.rdial contractility mediated throu6h beta adre.n6ic mechaai.ms. This increase in contractility is attenuated by secondarY stimulation of pulmonary inflalion reflexes. Aaordia6ly, wpep the hyperventilation that occurs with carotid chemoreceplor stimulation is prevented, the isotropic respnre Is significantly 6reater. Fisally, (or an equipreaaor sespore, the caro/id cbnsro- reeeptor reAea elicited a greater increw in contractility tha* did the carotid b.roreceplor reflex. Yaner. S. F. and Ruthertord, l. D. lownl o/ Cpstr.f lnreitl`arlon 61(6):1593-1601, 1978. Oth.r arrprir U. S. Public Hedt6 Servies. Frwa the DepartmeN of Medieiee, Harvard Medical School and Peter B.M Bri6ham Hospital; the DeparuaerN of Cardioloq, Childrenl. Hospital Medical Center, Boston; and New England Regional Primate Research Ceeter, South- boro, Mw. THE RELATIONSHIP OF REGIONAL CORONARY BLOOD FIAW TO MITOCIIONDRIAL FUNCTION DURINO REPERFUSION OP THE 1S('l1EMIC MYOCARDIUM , The purpose of this inveNiptioa was to relate changes In reRioaaa ooro.ary blood flow to the type and degree of biochemical disturbances that occurred during oc~haion of branches of the left anterior descending coronary anery and during reperfusion of the occluded area. To make these nwasuremenls, two groups of dogs were estsblished: group 1. moderate ischemi• before retlow, and group 11, severe ischemia prior to reAow. Using labeled microsphores, regional coronary blood flow was determined before liption, alter 60 mia. of 35 I

1-h , . . ischemia, and a(ter 15 min. of refbw. Hearts made ischemk for 60 min. hut not reperfused served as controls. aroups I and 11 were distinguished by several fealures. Oroup 11 showed a marked exacerbation of biochemical damage on reperfusias of the ischemic region (reduced levels of ATP, im- pairment of milochoodrial oxygen consumption and mitochrondrial calcium binding). Thia was accompanied by dgnificant aubendocardial hyperemia. Reperfusioo in group 1, on the other band, partially reversed these changes. Milochondrial calcium uptake aad ozidadve pbosphorylatioe (ADP/O ratio) were nol aAecled in any group. Tbeae data illustrate that the degree of bio- chemical damage following reperfu.iost of Ihe iachcmic myocardium is de- Iermined by the degree of isdsemia, sid suggest that interference with ATP productioe by the mitochroodria is aot responsible for the damage. Weiahaar, R., Tachurtscheotbakr, O. V., A.hiRawa, K., and Sinr. R. l. Cu.Jlofory 64 : )10. )64, 1979. Other erpport: The Hoover Fouodalioa From the Hualinpon Institute of Applied Medical Research aod Huntington Memorial Hoapilrl, Pasadena; and the University of Southern California, Los Angeles. TIIE EFFECT OF ALCOHOL ON TIIE HEART lo this very thorough review, the effects of skobol on the beari are examined In several different ways. The ftve maio sections of this paper are devoted to: (1) Historical Considerations; (2) Clinical Features; (3) The Acute Effect of Alcohol on Coronary Flow and Myocardial Performance; (4) The Acute Effect of Alcohol on Cardiac Melabolism, and (5) Correlation of Function and Structure. From a clinical standpoint, alcoholic cardio- myopathy cannot be differentiated from other myocardial diseases unless a history of akoholism b documented. The dinkal course of the disease depends on the duration of iaeAriety, aod abstiaenoe /rom ethaool is the most imponaot therapeutic consideration. lienwdyoamk studia show that the changes in- duced in animala aad men by prdooged perioda of ethanol administration do not differ from those observed is other types of eardioreyopathy. However, akohol allecta other ordaro, primarily Ibe li.er, and the bemodyoamic changes depend on the absence or presence of hepatic disease. The changes Induced by ethanol oa cardiac metabolism and fuaetion result tran its toxic action on the myocardial cell. However, all Ihe ooted changes ioduced by this agent an iodiuin6uishabk from other eardiomyopathia. In man, leakage of enzymes as well as of potassium and phosphate from the hean is observed. Ethanol also causes diminution of respiratory function of mitoehotdria and reduced activity of iruramitochondrial isocilraN dehydrojeoase. 11 dimirinhes calcium uptake and binding by the aarcoplasmk reliculum. 11 is also likely that i1 damages the contractile proteins. Most Irnportantly, a close correlation exists between function and structural ehanpes induced by ethanol. OveraN, much evidence exists that alcohol aQects a8 aubcdlular Mructurn, rewlting in their bio- chemical and biophysical malfunction. Bint. R/. and 1 illmanns, H. 36 I In: Lieber, C. S. (ed.): MdaWk Alpects o/ Akolioltsm, Laecaater, England: MTP Press Limited, 1977, dsapt. 4, pp. 117-134. Other support: U. S. Public Headh Service, Tbe Hoover Fouodatioo, and the Norris Fouodation. From the Huotingtoo Memorial Hoepild, Huntington Inslaule of Applied Medical Research and the Califoraia lns1ilule of TocbrwlOpy. Paudena; ad the University of Soulbero Cdiforoia, Los Aagek.. THE EFFECT OF ALCOHOL ON ACTIVE AND PASSIVE STIFFNESS, AND ON ISOMETRIC CONTRACTIONS OF OLYCERINATED HEART MUSCLE IN RATS Accordin4 to the eaperimew reported hen. the beuta of rata exposed b alcohol show several ooncurreal changes: shorteaisid velocity at zero bad (V..), total torce (P,) aod maaisaf rate of Ietrioa developmesM (dp/d1..) were all reduced. Tbe lirae oeeded to reach peak 1e..iow (1.) remained tbe same. As was found with fresh papitl.ry nMnele, the el.stkity module of active plyceriaated mu.de iacrea.ed lo proportioa to luad. 'ibere was rd- t.ilkmt elevation also in the t1iQ.esa of the seria elastic eksmeM. Ilo.re.er, no significant difference in res/i.d statu pessive tNiAaiesa waa observed betweeo rata exposed to alcohol asd the .os><paed controls. Other yet unpublfsYed results also indicate aorm dsnya io dn neliw aliAneas of infarcted beart mmck. The «lalio.ship ot aM Iheac data to the developmea of snyocudial failure rem.ros to be ioreslipMed. Maruyama. Y., etnr, R. /., Sarns., l. S. M. and Weishaar. R. lepestze Htart lorrwa/ 19(4):S1)-321, 19711. Other arpperts U. S. Public Health Servia` The Hoover Foundatio., and The Wright Foundation. From the Iluntingtoo Institute of Applied Medieal Research and HuatiWoo Memorial Ilospilal, Pasadeaa, Cal.; ud the University of Southern Califorda, Los Angeles. EFFECT OF ALCOHOL ON THE CONTRACfILE AND ELASTIC PROPERTIES OF GLYCERINATED HEART MUSCLE FROM RATS The effect of chronic alcohol coroumptiow on (1) the contractility and (2) the active and passive ali0nesa of the rat hearl muscle was examined because i1 had been fell thal changes io these p.ramelen might herald Ihe development of alcoholic cardiornyopathy io eaperimenld animals. In the lesu reported here, passive kngth•temion relationship in testing uale, iaoroetric contraction and isotonic quick «kase, and sti/fnesa of series elastic ekmenl (ataive stiRness) were measured in glyeerinaled heart muscle 1lbera obtained from 13 Sprague•Dawky rals maintained on alcohol tor an average period of five weeks and from 12 correspondind controls. Results show that is the hearts of rats exposed to akohd, the series elastic, as well as contracuk, elements are impaired. The rate of increase of active stdlnesa with tension 37

is signifkaotly higher for. akoholtsposed rats than for controls, while there is no significant differences in the stiffness at zero tension. This could indicate that in akohol-esposed hearts the changes is Ihe series elastic element con- tribute signifkantly to the deterioration of the myocardial penormance at higher loads. The observed akohol-ioduced diminution in the value of V~,, which is a measure of the rate at which mechanical energy can * produced by the contracting rnusck, also isdicales that the akobol-eapoaed myocardium cannot meet higher demands in energy output. While it is premature to attempt to esplaie the underlying meeha.ismy treae results indicate that the increased sliRoess of the seria elastic ekuweN awd the decreased contractility of the cardiac muscle induced by chronic akoAol admieiuration might play an im- portant role is the pathogeeesis of akobolie eard'aenyopatby. Maruyama, Y., Sarma, 1. S. M., FiscMr, R., Bertuellia, S., and elwl. R. l. In: Seisas, P. A. (ed): Cwrints /n AkoAo/bwr, New York: Orune i Stratton Inc., 1978. vol. 111, pp. 393-404. Otber supportr U. S. Public Health Service. The Hoover Foundatioe, and 1be Wright Foundation. From the Huntington Institute of Applied Medical Research, Huntington Memorial Ilospital and the California Institute of Techoolop. Pasadena; and the University of Southern California, Los Angeles. CONTRIBUTION OF MYOCARDIAL METABOI.ISM TO T11H KNOWLEDGE OF MY(X'ARDIAL FAILURE AND MYOCARDIAL INFARCf1ON Myocardial failure and myocardial infarction are the most frequently seeo acule cardiac disorders Increased understanding of cardiac metabolism now provides greater insight into these probkms, their cause and, therefore, the therapeutic approach to be followed. Myocardial failure is defined and dis- cussed in 1he light of contractility, cakium transport and uplake, and the role of milochoodria, all of which are disturbed. Myocardial infaretion, on the other haod, is defined as regional i+chemia, and a knowledge of anaerobic p.Ihways controlling the biochemkal events that then take place becomes essentid in order to rodentaed them. Here, tbe rate-IimNlng ensyma are really the ton- IroRiej tacbr+. Fatt)r acid metabolism is significantly altered, as are IM gIycotylie cycle intermediates. Whether or not all Urese concepts can be translated isNo therapeutic applieatiorr is still uncertain. What Is ekar, however, is that myocardial failure and iofarction can sow be studied by a new approach stemming from the esamhnatioe of fundamental mechanisms In heart muscle instead of relying solely on clinical observation. Sint. R. l. Recent Adr.ncrs /e Studies on Cardiac Structure and Metabolism 12:3-9, 197f1. Other supportr U. S. Public Health Service aed The Hoover Foundation. From the Iluniintlon Memorial F1inpital and Huntingloe Institute of Applied Mahaal keseuth. h+adeaa; and the Ilnivenity of Soulhern ('alifornia, tos Angeles I IV, deuroplr.rmocolosy.nd Physiology PSYCHOBIOI.OWCAL CONSEQUENCES OF CHRONIC NICOTINIZATION: A FOCUS ON AROUSAL While nicotine has been discordantly depieled an haviaR tr.nquilisjoR, slimulatiog, depressios, qood-stabiliting, and Mrwatteswatio8 propertiek there is one feature commoe to these various categories: Wey dl can be related to an orjanism'a qate of arousal or activation. 1n this paper, nii:otisei ad=vns in animals are characterized in terms of the nwdibcatioes it can affect in the kvel a.d/or the aualitative nature of arousal. Earlier behavioral aed ekctroenceph.lographic works ars reviewed, and recent etudies of niootise's apparent aMarnisun of both neurophysiologiealy-isdueed and environmentally- induced slates of hyperarouaal are preaeated. Careful attentioa is paid to the diAerenees between acWe and ehro.k Bicotinisation. asid the rok of tokra.oa is closely acrutinised. Since the rdationship between iaherent k.ds of emo- tionality and the readiness with which tau enpBe in consummatory behavior had beea previously eslablished, possible interadioas between baseliee easa tionality kvel, environmental stress and nicotine administration were co.- sidered. Resulu showed that chronic uiootiniralioes produces an antagonism of the effects of environmental strese. iber also gave evidewoe of a more effective protection by nkotiee for animds with relatively higher inborn emoliooality levels than for those with ksw<r ewsotioeality levels. These eaperiments M- votvinR maoipulatioro of arousal fevds suggest etut nicotine caw indeed i.buoa a behavioral state which i less wroeplibie to distuption by intrusivr environ- rneetal or intersial ehalknges. Nelsee, 1. M. (Goldrteiw, L.) Ie: Brittis, K. (ed.) : AeA.rbral Eaects o/ Nkadne, Basei: KarBer, 1978. pp. 1-17. From the Department of Psyehfatry. Institute of Mental Health Scie.us, CMDNI-Rutgers Medical Scbool, Psealaway, N.1. CEREBRAL UPTAKE OF NICOTINE AND OP AMINO ACIDS Nicotine enten the brain from the circulation very rapidly; it also ir creases blood fiow. Since this rapid uptake of nicotine indicatea a nwds of transport different from other melabolic proees.es, such as those for amino acids, it seemed important to study the mechanism of nicotine uptake. In order to esamine the possible interaction between the uptake of niastine urd amino acids in mouse brain ln viro, capillary transport was measured by comparing the passage of 'Y'-labekd compound to a('H) water reference from the capillary circulation into the brain; !w vitro studies were also done by measut- ing incubated dices of mouse brain. Results of Ihese and other eaperimeota showed that nicotine, which enters rapidly, also leaves the brain rapidly; an eait somewhat slower than Ihal of water. /n spite of this rapid eaiN, brain levels remain higher than blood levels over a period of time. Nicotine enters the fetal circulation from the maternal blood; it enters fetal brain but to a smaller 1" 39 '

: y estent than adult brain. Most close analogs have no effect on nicotine uptake. Nico- tine has no eAect on amino acid. Rlucose, or nicotinic acid uptake in vivo, indicatinR that nicotine does not alter upillary or pericapillary metabolite transport s.od does not itself participate in such trampott processes. At higher coocentrations, however, nicoline uptak. Y.aturabk. Since nicotine uptake does not follow the properties of active transport systems, its uptake probably represents binding. Nicotine and morphine mutually inbibit each other i uptake. These resulla suggest an uptake compartment (lipid space) for nicotine shared by morphine. Senhen. H. aod L.JrAa. A. Journal of N.woicknce Rrrr.rA 1:tf3-91, 1979. From the Center for Neurocbemistry, Roeklaed Research Intlitule, New York. THE EFFECT OF NIC.OT7NE ON THE METABOLISM OF BRAIN PROTEINS The inteot of this study was so iovesligate the effect of nicciine on pro- tein synthesis in the adult and immature trwuse brain by meawrint amino acid iocorprxation !n vlvo and /n vitro. Results iadicale that nicotine can influence cerebral protein metabolism. In vitro incubation of adult brain slices with nicotine stimulated valine incorporatioe into protein. This effect was specifk to t.-nicoline; bnicotine decreased incorporation, while nicotinamide and nicotinic acid were Ineffective. Animals treated with nicotine in vivo showed a tendency toward greater incorporation but the Increase was not signilkant. In the im- mature brain, nicotine did not have the same effect on protein synthesis as in the adult brain. In the newborn, it significantly iohibited valine incorporation both in v/no and /n vivo. The variability of the results obtained with adult brain tissue wtgesls Individual differences ie nicotine sensitivity. The decrease in protein synthesis observed in the newbors may aupport previous evidence that neonates born from animals treated with nieotine during pregnancy en- perience delayed devebpment. However, this may not be a long lasting eAect, since no abnormality is then found in the adult tissue. On the basis of these data, it b thought that, in the imrnature brain, control aod pharmacobiic sensitivity of proleis metabolism are not the same aa in the adult brain. Senhen. H. aod L.jtA., A. Nerroabarnacototy 1d:76)-766, 1979. From Ihe Center for Neurochemisiry, Rockland Research Institwe, New York. Ft FCI-ROPHYSIOI.(X:ICA1., BEHAVIORAI., AND CHEMICAL I: .VIDI?N('E F(1R A NON(-FIOI.INER(iIC, STEREOSPECIFIC %I11? 1()R N/( tt11N1' IN RAI' BRAIN M.r....aJ h.,t .r sn .nrm(•r t.. Jcmoosuste that crrtsur Inain reRinns .,.., h... n...t..a.n..s. ,,..w,rr rnry-I"r s,Irt end that thc.c aue at lea.t I 100 times more respomive to the natural (-)-.icotine isomer tha* to the (+)- one. A speci0c beiiavioral response to nicotine administered into rat lateral ventricks is described, and correlative ekdrophyaiologic, psychopharmacolosic aod «uptor-binding data are furnished b wwpport the contention that tha ailes are noncholinerRic. The iojcetion of 2-10 tog of (-)-aicotine bitartrat. ioto the lateral or third ventricle induced a prostration-immobilization sy.- drorne. A dose at least 100 tirnes larger was required to produce th, sama effect with the (+/-bomer. A number of other agesNa were tested tor tbeir ability to prevent this aysdrorne, including a series of synthetic oieqtiae and piperidine derivatives- The beezyl or 4-mido-2-nitropbenyl derivatives of either compound (10rR) admis>iuaed iotraventeitarlarly two mitwles before 4 rR of (-)-nicoline prevented Ihe syndrane. A wide variety of traoquilisera, neuro- lransmitters and their agonius, u wep as cholioerRic agents failed to either sire- utate or block this nicotine effect. Elcetrodn implamed in the dond hippo- eampus of freely moving rata recorded diuirret eltettophysiologic p.tterm associated with the behavioral tespome. 71ese mainly ahowed a decrease is the frequency of impuMe and shifls in amplitude. A number of aynthelk nicotioe and piperidine derivatives also iohibited the electrical effects of nioo- tine. Whik the binding of aH nioo(iae to netual rnembr.ae preparations could not be demonstrated, the slereoipecific binding of nicotioe in fresh braia slica and to Rlass fllten was. For Rlass, the K, was 2.0 X 10-a. Af ood, L. G. er J. lournal of Newozcitace Risea.ch ):327-)7l, 1975. Oth.r aupportr U. S. Public Heakih Servioa. From the Center for Brain Research and the Departrneot of Biochemistrp. U.ivenity of Rochester Medical Ceeter, Rocbesler, N. Y. EVIDENCE FOR A NONCHOLINER(3IC SITE FOR NICOTINE'S ACTION IN BRAIN: PSYCHOPHARMACOLOdICAL, ELECTRO- PHYSIOLO(jICAL AND RECEPTOR BINDING STUDIES Although much is known about the psychological aed pharmaooloRits.l effects of nicotine Is man and anirnab, the brain sites and mechaaisrm of action of nicotine are still only poorly understood. In an attempt to rectify this, the present report describes a variety of Madics aimed at investiptinR the esistence of noncholinerstic siles for nicoline i action within the rat's brain aad using townds Ihis ubiective a newly-developed Rrcwp of antaRonists to the behavioral effects of aicoline. Injeclion of 1-10 pg of (-)-nioWiee Into the region of the lateral ventricle of rats through implanted eannulae tewlled in a characteristic proseratioa imnsobilization syndrome, which was accompanied by seizures and Iremors a1 the higher dose range. 7 he ( t)•isorner possessed 1/I(!0 the activity of the natural (-)•isomer. This syndrome could be pre- vented by prelreatment, intraventricularly, with Ihe N•benzyl anJ N p-niuo- phenylazidu derivatives of either nicutine or pipetidine. A variety u/ neuro- transmitlers and psychotropic agenls, including aeelylcholine and antichotinergic dtur, wete without antagonistic action Recep(nr binding studies were per- 41

0 rV 0 I I i formed with rat brain slices and varbus seural preparations using 'H-nicotine, 1„1-orbunprotoxin and 1'C-d-tubocurarina as liganda- oodr with 'H-oicotine was It poasibk to demonstrate any competitive effect with the various oicotioe and piperidise antapebts. 'ibese resulb, in c,oojuoctioa with the data ob- taiercd frorn recordinp of spontanean ekctrkal activity, suggest that speci0c noe-cholinergic sites for nicotine action exist which have not been hitherto described. A6oo1. L. G. et d. Arclilr.z Iwi..nnrlondrs 4 P/u,nuealYn.ndt er l. Therep1.237(2):213-229, 1979. OtAer w'prtt U. S. Public Health Servioe. From the Center for Brain Research aad Dep.rtmeeA of !liochemistry. University of Rochester Medical Ceeter, RocheNer, N. Y. RELATIONS BETWEEN NICOTINE-INDUCED CONVULSIVE BEHAVIOR AND BLOOD AND BRAIN LEVELS OP NICOTINE AS A FUNCTION OF SEX AND AGE IN TWO INBRED STRAINS OF MICE Blood aod whole brain nicotine kveb were determined ie C3: sU61 and DBA/21 mioe following intraperitoaeal iejeeUon of nieotine, and their relatio.- ship to eertain behavioral effects was examined. Speedkally, the LD„ and ED , t/ahrcs were established for each geeotype, ses, and sye of the animals atuaied. These were then compared to the blood .nd brain nicotine levels i. order to determine whether the behavioral effects of this alkaloid were related to the levels found in these tissues. Blood levels were also compared to brain levels to determine if the former relketed the latter accurately enoukh to permit monitorio6 brain kveh by analyzing smaR blood samples. (Such a techniQue would maintain the anirnal intact for long-term studies of the behavioral and pharmacologie effects of nkoline.) Resulls indicate that blood levels alone are not sut iciently accurate to predict either the brain levels or the behavioral respon.es resulting from a single injcctioa of nicotine. However, brain levels were generally fairly accurate predktors of the behavioral respooses. According to the data, male and female mice differ in their sensitivity to nicotine. Maks ol both strairo (42 days old) givea oomparabk doses of nkotine showed a higher brain concentration than females of the sanx age. There was no cwret~ ponding increase In sensitivity to the increased brain eoncentralion, however, as measured by the I.D,,. EI), or latency to tremor or death. Tepper, 1. M., Wilson, 1. R. and Schksinger, K. (McCleern. G. E.) PAoinecolop QiochnnJirry d daA.rlor 10(3):319-)33, 1979. From the Ihpartment of Psychology and Siobgy, and the Institute for Be- haviural (ienetK+. University of Colorado. Boulder. i I t MA7VRATIONAI- CHANOES REtaTED TO DOPAMINE IN THE EFFECTS OF !-AMPHETAMINE, COCAINE, NICOTINE, AND STRYCHNINE ON SEIZURE SUSCEPTISILtiY lo this examination of amphctami.e: eQeeta on convulsive behavior, tb actions of several neural eacilanu were contrasted with Uwaa of aarphetamiaa. Tbe agents chosen for study had diverse nwdw of actios and We greatest likelihood of differentiating mpoclonk and cbok .eisure.. For Ws re..oa, the effects of 1-amphetamine, cocaioe, niootiawr, aad strychnine on drese two parameters were investigated In two age Lroupa (30 and 120 days) of ahorl- skep mice. Amphetamine and cocaine decreased stroeptibitity to ayocloswr is young mice and Increased ausoeptibilily in mature mice. 'ileae effects wera attenuated by pretreatment with haloperidol, iedicatins mediation by a dopantimer6ic system. Amphetamine did not ailer elook suaceptt'biliry in either age group of mice, whereu cocaine affected elonie susceptibility and myocloaru. Theee effects were reot attenuated by h.toperidd, iwdkati.6 mediatioel by systema other than dopemine. Nieotis decreaaed uacxptibility to myoclonr and incre.sed susceptibility to clorws, whers.a Mryeb.ins increased .v.oep, tibilitp lo both types of seizure. Haloperidol, howerer, failed to alter aay of these effects. Tbese results an eoasbleot with earlier wrork, which suyeste that a dopaminerjk mechanism in thesa mice trderRoa marked developmental ~ change between 30 asd 120 days of ye. Oreer, C. A. and Alper., H. P. (McCk.n., O. E.) P+7cAolArn..rofogy 64(3):233-260, 1979. Other e.'prtr National lestitute of Mead Health. From the Department of Psycboloq, aaid the Itrtitute for sehaviord (kwetks, University of Coiorado, Boulder. SEROTONIN IN SEIZURES AND SEIZURE DISORDERS In this very thorough review artkk. reirrn aetivity aad overt oonvubion are carefully eaamined in relation b brain twttolad.. Specifica111r. ura.y ea- perimental and clinical atudies focusing upon asrolonis r eitber a causative molecule or a reflector of change is relatioe to aiwrs activity arm praealed here. Major sections of the paper deal with: Changes in Seroto.ie with Cors- vubionr Monoamine Oaidase; Drug Intetactiona; Enviroenrenld leiteractioee; An/konvulnaMs; and Cerebrospinal Fluid. Individual studies Indicate that eonvubiom can be induced by electrkd stimulation, chemicals (tiuorothyl, pentyknetrard, lidocc.ine, 3•hydroatiryptophar, hyperbark oay6enatioo oua- bain, tunptk acid, cobaM, plhenelzine), and auditory etiniulalio.. OveraQ vari- ow esperimentd Ondinp show that the convulsion produced by electrical stirw- ulation affects serotonin level ard/or metabolism is the whole brdn, regionapy, cellularly and subcellularly, depending upon the characteristics of ewreat flow and of 1he seizure activity produced. Serotonin changes appear to be furUw regulated by such factors as age, environmental and drust-indueed alteratloes, the interval after convulsion at which changes have been measured, and the J W 42 43 ~--~

r number of convulsions that have preceded such measuremenu. lust as the occurreoce and nature of brain serolonin changes produced by electrical stim- ulation may differ according to speeies, brain regions aod sites of electrode placement, there also appear to be differences In the relationship of aerotonin to aizures produced by various chemical agents. It is pointed out here that the intent of this chapter was to formulate a series of relationships between brain serotonin (its concentration, metabolism and physiological role) and the e:ist- ing evidence Ihat relates these fac/on to the elucidation of seizure mechanisms, regulation of seiture activity and the potential Implications that they may have for the diagnosis and poaibie trtatmest oI seizure disorders. Eunrow, W. B. t In: Esaman, W. B. (ed.): Serorenln fn Health and Dlrcase. Vol. llt: The Cen- rrd Ncrvorr Syrrrrn, New York: Spectrum Publ., 1978. pp. 317-401. From Queens College of the City University of New York, Flushing. . TiiE F.FFEC7S OF NICOTINE ON IMPLANTATION IN THE RAT Recent studies demonstrate that nicotine administrHion significantly re- duces uterine re+ponsivity so decidualicalion. 1fLis particular report eaamioea the effects of nicotine adminislralion on Implantation In the rat. Pregnant ani- mals were injected twice daily with 5 mg nicotine/kil body weight from the first morning after mating (Day 0 of pregnancy) to Day 5 of pregnancy. Corte-. spondia= coMrob received equal volumes of saline. By flushing each uterina horn with a constant volume of saline under constant force at various Intervals during prepnancy, it was established that implantation had progressed su/ll- cienl/y by 1200 hr of Day S to prevent eonceptua retrieval in control animad. In the edcWine-treated animab, however, blastocysts were retrie.ed as late as 2100 hr on Day 3, indicating that attachmen/ suAkient to resist /huhing wes delayed for as mueh as nine hours. This delay waa sot attributable to altera- liora in oviductal transport, since the fertilized ova entered the uterine )umea at the sanw time (1800 hr on Day 3) in both control and treated animals. Delayed blstocyst development waa noted in the dcoline-trealed rab: 1he time of zoos pellucida loss and the growth of the Inner cell man were delayed. NeverMeksa, the number of blaatoeysls ultimately implasled was the ume as In the controb. While Ihese eaperiments clearly establish the ability of a high dose of .icotine to subtly affect certain reproductive mechaninns In the rat, they do nw resolve whether the e/Iecta noted her. «suk from the action of the alkaloid on: (1) the hypolhalamo-pituitaryovarian aair (2) the conceptus; ()) the uterus: or (4) the blastocyst via the u/erw. Further eaperiments should determine eaactFy how nicotine alters blastocytl-uterine interaction in the rat. Card. l. P. aod MltrAcll, J. A. dlology o/ ReprodrerJon 20:372-579, 1979. From.Ihe Ikpartment of Anatomy. Wayne State University Sclwol of Medi- cioe, Iktroit. I HORMONAL CONTROL OF IMPLANTATION IN THE RAT: INHIBITION BY LUTEINIZIN(I HORMON&RELEASINO HORMONE AND ITS ANALOOUES Reduction in the nnourtt, or el'iminalion, of either estrogen or progesleroaa delays or abolishes ovum implantation in the rat. Thie paper presents reoed observations on delayed implantation e:perimeolaMy Mduced by luklnizing; hornwne-rekasing hormone (LH-RH) and ita analogues in the hope of deter- mining some aspects of the underlying mechanism controlling the sequential hormone secretion responsible for ovum implantation. The inbibitor)i activities of Analogue 1(des-t31yr•.LH-RH-elhylamide) and A.abpse 11 (des-Olyr' (D-Ala'/-Ll11-Ril-ethylamide) were 4.5 aod 173 dmes greater, respectively, than that of the native hormone. On day I of treatment with Analogue 11, 1he pituitary content d/J/ and PSH In pregnant rats waa about 10% of that found In untreated pregnant rats. Ilowever, the serum levels of 1.11 were significantly hiRher, and those of PSH siSni/kanUy lower, is the treated rats than in the controls. The Andogue-11 Ireated animah aho bad lighter ovaries than the controls because of smaMer corpora hNea and kss developed follicles. All three compounds signi/kantly lowered peripheral serwn progeslerone eow ceatrations, but the native hormone and Analogue I brought this about sooaer than Anabgue 11. Concurrent treatment with Me following hormone combi- aationa blocked the Inhibitory effect of Analogue Il: progesteroae + estradid; prosesterone + human chorionic soa.dotropin (hCO); prolactin + estradid; and prolaclin + hCO. Neither progesleroae not pr dactin alone had any elled. These data indicate that Analopre II alinwlales the production and teh:asa of LH and the release of FSH, increasing ehe ratios of LH:prolactin eod LH:PSH. 'iLe gonadolropia imbalance thw induced suppresses the development of e~var- iat follicles and corpora lutea and reduces 1he secretion of both estrogen a.d projeslerone, perhaps by altering receptors for steroidotropic hormo.es. This diminidrcd secretion, in turo, alters tNeriar receptivity to implanlatio., reauh- ing is its inhibition. rosAin.pe, K. and Pujino„ M. , /.: df.ternd RrcotnUlas of rretwracr, tib. FouadNioa Series 64 (tsew sa- id), New York, AaWerdsn: Paeerpt. bledice, May 1979, pp. t3-103. From the Laboratory of Humaa Reproduction and Reproductive Biolop, a./ the Department of Anatomy, Harvard Medical Sebool, Boston. EFFEC'fS OF NICOTINE ON EARLY PREGNANCY IN THE RAT Since no studies have bees reported on the effect of nicoline on. preiaa- plantalion embryos or on hormone levels during early pregnancy in th. ra1, a very high dose of the alkaloid~waa used In lhis pilot study b select para- meten potentially vulnerable 1o nicotine for later investigation of low dose rAects. Accordingly, a 7.5 mg dose of nicotine lanrate was injected subcutaneously Iwiee daily from the morning of proestrua (Day 0 of pregnancy) until the day of sacrifice. Rats were sacriflced a1 600. 1200, 1s00 or 2100 hr from Days 1-S of pregnancy and the location and stage of development of the ova were recorded. Results show that Ihese daily injections of nicotine eserted a variety of effects on the embryos and on the genital tract. These effecu are w li N 44 43•

I I (1) a 12 hr delay in embryonic cleavage from the 2-cell stage to the 4-cell uage, (2) delay in the entry of embryos ioto the uterus, (3) del.ty in ovum implantation and (4) alteration of ulerios movement respoesibk for spacing of the ova. A single injection of 0.2 pg estradiol on Day 3 of pre4nancy abol- ished the delay in ovum entry.inlo the uterua, whereas daiy nkotise injection caused a concentration or "crowdinj of implantation sites toward the lubal ends of the uterine horm. Conoeetratioos of progesterone, hNeieuinl hormone and prolacde In peripheral serum during the preiaplaetation heriod were lower in eicoline-treated rats Ihae in saline-treated ra1s. On the other haed, serum concentrations of estrogens and foMicle Nimulating hormone were higher ie ekoline-treated rats than they wen in saline-treated rats. These results sut- gest that delayed implantation of ova in ekoline-Ireated rats is due not to a decreased secrelbn of eslrokee, but rather to a delay In Iht iecrease of proges- teroee which is needed to prepare the Wtrua (or implanting bla.tocysts. This disturbance in propesterone aecretion appears to be the result, at least in part, of a hormonal imbalance in tbe hypothalamo-peuitary aais caused by the nicotine treatment. YorAlnagq K. et al. Sloloty of Reprodretion 20:294-10), 1979. From the laboralory of Human Reproduction and Reproductive Biobgy, and the Departments of Anatomy and Obstetrics and Gynecology. Harvard Med- kal School, Boston. EFFECT OF NI('UTINE ON TIIE DEVELOPMENT OF FETAL AND SUCKLING RATS Recent evidence suuests a causal relationship between maternal smoking and babies of lower than average birth weight. This investiplion attempts to determine the effect on fetal development of exposure to cigarette smoke or nicotine administration at levels comparabk to smoking by the mothers. A nicotine dose of 100 pg/kg/day administered to rats starting on day 14 of gestation did not affect matereal food intake, weight pin, length of gestatioa, liller sae or fetal development. A daily dose of I mg/kg, however- did reduce litter size and produced a higher thae normal iecideecs of stillbirths. Con- tinued nicotine administralion so the mothers of 100 pg/kB per day until 12 days postpartum did aw affect newbore body weight and kagth, or heart and lung sise during the first week. In the second week, however, the newborns of ekotine-treated mothers did not progress aa rapidly as those of controls. Their stomachs contained less food than those of the control group of new- borns. iltia diQerenee increased with qe, until it reached more than 40% t 12 days. It is suggested that nicotine probably inlerferes with normal milk productioe, thereby affecting the development of the offspring during a period of rapid growth. HanrosA, M., Simon, M. R. and Ilamosh, P. Rlology of the Nronnte l3 ( 3):290 297, 1979. From the Department of Physiotop and Biophysks, GeorSetowr University Schools of Medicine aod fkntiury, Washington. D.C. 46 I I THE RELATIONSHIP OF Q-BUNOAROTOXIN BINDING ACTIVITY AND CHOLINERGIC TERMINATION WITHIN THE RAT HIPPOCAMPUS Light tnicroscopy autoradiopraphy was esed to study the distribution and behavior of w-bunproto><ia receptor sitea withie the poNealal etd edult rat hippocampus. Histochemical visualization of aoetyk hdienteras. (ACSE), andd amino acid tracer techniques determined the temporal and spelial relaNonshipa of toxin receptors and presynaptic cholinergic ekmenb. Several observations were noted as a rcwM of these experiments: (1) Tonie-bindiait sileap develop very early, probably before the ingtowth of cholinergic Abers. (2) Toxin re- ceplors develop normally in the absence of presynaptic inAueects, as denae- tlraled by septal lesions whkh destroyed the cholinergic pathways. (3) Two arar of marked bain-bindiag activity lie outside regiona of identi/kd cholieer- Sk termination. One of /hese is in the hippocampus of the eewbore, where receptors appear uneapteledly and ttansknly. The other ia intriguingly found in the adult hippocampus M an area apparently devoid ot cholinerBic lerndaa- tion. These data lead to the coecheioe that, in the rat hippocampus. .-bu.- pro/oaie binding sites, which are likely to be relaled to nicotinic acetylchdiea reup/ors, develop independently of eholiecrRic Iapu1 and may be involved Is nonsynaptic traesneuroaal interactions. Thia type of activity may reflect the action of acetykhdine as an eatraryuptk neurolwrnoral atbstance instead of as a conventional synaptic transmitter. Huet. S. and ScAnridM, J. Newoarknce 4:3l3-392, 1979. From the Department of Psychiatry and Behavioral Scknce and the Depart- meet of Biochemistry, StaM University of New York at Stony Brook, Stoey Brook. PUTATIVE ACETYLCHOLINE RECEPTORS IN IIIPPOCAMPUS AND CORPUS STRIATUM OF RAT AND MOUSE While the central nervous system of the rat has been extensively studied with respect to the location and characterir.ation of nicotinic and muscarink acetykhdine teceptots, little previous attention has been paid to these receptors in the mouse. In this sludy, the anatomical distribution of „-Bunprotoain (.Bu1X) binding sites in the hippocampus and corpus slrialum of CS78L aed 1)OB adult mice was compared to that of adult Sprague-Dawky rats. Auto- radiographic and hiuchemkal measuremenls determined the level of aBu7X and QNB binding in thcae two atels. 'fhe autoradiotlraphic study reveakd ae almost compkte absence of labcl within the caudale-putamen of the rH; in conlrasl. the strialum of the mouse was rather heavily labekd. This obstrva- lion was born out by /n vitro measurements of e8ul X and QN8 binding activity. The QNB binding levels in mane and rat did not di/ler, bot si4nifkant di/ferences in the conceneralron of .Bu1X siaes were found in the striata. lhe differences in the concenlralion of nBti IX sites in ra1 and mousc are not limited to the slriatum; they have been observed alw within the ral'a 47

cerebellar granule iayer, where there are none in the mouse. In Ihe ral, the high levels of (?NB binding activity in tbe striatum seem to indicate that acetykholine receptors are primarily of the muscarinic type. Neverthekss, there is evidence for the esistence of a few nicotinic receptors aho. In the mouse, considerable information on drug binding properties and analomicai distribu- tion indicates that the Bul X receptors may be nkolioic acetykhol,ne recep- tors. The iMer-species differences noted then migh1 mean that in tle mouse, nicotinic receptors fulfill functions subserved by muscarinic receptr.rs in tbe rat and vice vena, indicating thtl Veat care must be taken in the estra- polation of receptor data from one speciea to another. This report emphasites the potential of the mouse as an eatperimnNal animal in receptor rrearch. Marchand, C. M.-F., Hunt, S. P. and ScAndh, !. drain Rerr.rcA 160:36)-)67, 1979. From the Department of Siochemistry aed Department of Psychiatry and Be- . havioral Science, Sute University of New York at Stony Brook, Stony Brook. INSECT ACETYLCHOI.INE RECEPTORS AS A SITE OF 1NSEC17CIUE A(.'IION While most of the current generation of insecticides are inhibilors of the enzyme scetykMobneslerase, this paprr looks at a different potential site of ac- tion of insecticrd.lly active mokcules, the pnstsynspticatly located acetykholioe receptors Ilcre. the bbnding of sn rsorheocyanate to an rnsecl acctykholrne re- ceptor was e.amined and cumpsred yu.nlualively with it ability to produce postsynaptic blockade 1 or cumpsrrxrn, parslkl e.penmenu were carried out using nicotine, which is both an insecticide and a potent ganglwnrc blockiog agent in insects. tAe agent ""'1-& bunprotoam, which binds specifically to nicotinie acetykhc>tine receplon of vertebrates, was used as a receptor probe. Results of these etperiments showed that 2-isothiocyanatoelhyltrimethyl-am- monium iodide produces half-maaimal block of rrssr1-.-bunprotoain binding to the insect estracts (PeriolenH. CNS and DrosoPhile whole flies), a reac- tbn that occurs at eoncentrations similar to those producing half-masimill suppression of escitalory postsynapt{c potential amplitude at a central synapse in Prrlpf.nra, which can be blocked by .-bunproto:in. Hence, this studr denanslrales that an Iawhiocyanate compound u+d nieotin., both of whkh show Insecticidal activity, are ationisls for an acetykhdine receptor in the CNS of the cockroach Prr(pl.nrra rnrrknns and stress the nced for detailed com- parative studies on the properties of insect and vertebrate choliner6ic receptors. Gepner, 1. 1., H.II, L. M. and S.Itelk, b. S. Nature 276:1tt-190, 1975, Other support: National Science Foundation. From the Department of BNrlogy, Massachusetts Institute of Technology. ('am- brrdee, nd rhe Departmenr u/ l.Kdogy, AR(' Unit of Invertebrate Chemistry and 1'hysndoty- I mbrwlRe. I nill.nd GENETIC VARIANTS IN AN ACETYLCHOLINE RECEPTOR FROM DROSOPHILA MELANOGASTER An experimental stratep for the detection of genetic variants that affect acetylcholine receptor structure is presented here. Nicotine hydrogen tartrHe in the medium at 3 mM was used to acreea for nicotine-resistant straios in wild-type DrofophUa populatiaa, and eight resistant strains were identi(ied. One confirmed nicotine-resistant strain, designated HR, was isolated from the llikone-R wild-type stock. In this populalioe, the survival rate wn 50% after esp.nure to the 3 mM nicotine hydrogen tartrate, which killed %% of the Canton-S strain. While sekction for nicotiee-resistant strains Increased the probability of mutations which affeet the nicolinie .eetykholine receptor stnrc- ture, not all nicotine-raistant nadan/s were expected to have altered receptors. In order to distinguish the ones that did, isoeketrie focusing of Ihe acetyl- choline receploe-Is"1-labekd rbunprobain complex was used. When the other seven nicotine-resislant strains were analyzed by this procedure, five showed profiles indistinguishabk from that of Canlon-.4. The remaining two uocka showed shifts is 'soekctric focusing point which may represent nmtations is the structural genes which code for the polypeptide subunits of the receptor. Sotb the nicol'ae-resistanoe phenotype and the alteration in i.oelectric point of the receptor-tosin complea that were observed in the HR and other nico- tine-retistanl strains could be due 1o alteration. in 1he structure of the aeetyl- chdine receptor. Irrespective of whether the same gene is responsible for both the nicotine-resistance phenotype and the shift in isoeketric point of Ihe re- cepor, the isoekclric focusing procedure presented hEre provides invealipton with a direct way to identify mutants with altered acetylcholine receptor structure. Hatl, L. M. e/ aJ. PEAS LETTERS 91(2):247-216, 1978. Othrr support: National Science Foundation. From the Department of Biology. Massachusetts Institute of Technology. Cam- ' bridge. V. P6.rmneofosy .nd Alocbensistry HUMAN I EUKO('YTF. t:1.ASTASE AND CA7NF.PSIN 0: ISOLATION, ('IIARA("1 L'R17A /1ON ANI) INTt:RAC1ION Wtl ll P/.ASMA PR(1TF.INASIi INHIBITORS . The polymwphonuckar (PMN) kukocyle conlains a hosl of degenerative enzymes which can be and are ulilised durinR the pha6ucylie process. Of theae en:ymes, at least 7% of the dry weight of the cells Is represented by col- IaKenase, elaatase and calhepsin (I. Lack of control of these prweinues by plasma inhibitors could result in Iissue dama6e. Because of their possible im- portance,  systematic study of the propcrties of Ihese en:yrnn, including structural, functional and regulatory asprcts, was surted. '11re resulls of this 48 49

study to date are summarized in this paper. A simple but rapid procedure for the isolatioo of both kutocyle elastase and calhepsin G is reported here. Using af6nity chromatography on Sepharose-Trasylol, followed by ion-exchange chromatography on CMcellulose, both enzymes were isolated in excellent yield (63% for elaslase nd 75% (or catbepain G). The isolated proteinases migrated as two distinct sels of isoenzymes with slightly diRererit ekctro- phoretic mobility, presumably due to variatiorn in carbohydrate conlent. Pre- liminary slrvctural studies on elasl.ae ad cathepsin G indicated thst the two enzymes are largely homologous lo each other and to pancreatic proteinases, but the still insufficient data do not yet permit any inferences regarding their mechanism of action. The protaolytic degradation of lung is also r.oosidered here, and close attention is paid to the interaction of plaama inhilitors with PMN claslase and cathepaio 0. 1s this laboralory, it wn noted that w•1- proteinase inhibilo. (.-I-PI) formed compkles with virtually all scrine pro- teinases. Furthermore, treNment of these oompleaes with nuckopluks, such as benzamidine, resulted in cornplat breakdown and the formation 4 a modi- fkd form of the inhibitor (.-1-PI•). Subseqoently, .-1-PI was obtained from complexes with etastase, cathepsin G, erypaiw, chymotrypain, thrombin and plumin, and In every case the amino terminal sequence was Identical. Tr.rb, 1. et at. In: llavemann, K. and lanuR, A. (eds.): Nernaf rrotearrs o/ Human Po(y- mo.phonrckor Ler4ocyra:, Baltimore-Munich: Urban & Schwarxenberg, loe., 1978, pp. Ile-12e. Otber support: National Institutes of Health. From the Department of Biochemistry. University of Georgia. Athens. MAPPING THE EXTENDED SUBTRATE BINDING SITE OF CATHEPSIN 0 AND HUMAN LEUKOCYTE ELASTASE. STUDIES WITH PEPTIDE SUBSTRATES RELATED TO THE Q,-PROTEASE INIIIBITOR REACTIVE SITE This report describes ,he synthesis of a number of 1-nitroanilide substrates for human kutocyte (t1L) etaslne,and cathepsin G.ed the various kinetic studies, which provide information on the nature of the extended substrate binding sites In these enzymes. It is currently thought that HL elastase and cathepsin 0 are Ihe aCenu responsibk for the destruction of the lung in em- physema. Evidence indicates that IIL elulase interacts with at least five resi- dues of a substrate (or inhibitor). Cathep.in 0 prefers negatively charged P, groups. Thb ensytne, along with many other serine proteases, cannot ae- - cept a Pro residue al its S, subsite. One of cathepsin Gs better subslrates. Suc-Ala-Ala-Pro-Phe-NA, is not hydrolyzed by Ht. elastase. These tools should prove useful in the study of the two enzymes' biological function. The best I-nitroandide calhepsin (i subsvate yet reported is MeO Sue-Ala-Ala-Pro-Met- N 1, us ns,d.ru.n vrrIJ% wn Juarrrumtrlc sulin..,fes Neither is IxrunJ lo, nr hvJr•dtr.J 1.v III rt.a.v -.r ..Ihtpun /i, auJ Iwah revat frMrrly with 1.-.,ra h h., p.r..wr,/t Irrn shu+u Ih.l o.iJ.UU/r u) ,., pro- tease inhibitor (.,-PI) destroys its inhibitory activity. Together with this observation. the data presented here aldicale that oxidation of the P, Met of .,-PI can destroy its reactivity toward oaN serine proteses, including the two major ones in the granules of human leukocytes. Thus, oxidation at the .,-PI proleolysis site could explain 1he inhibitor s inactivation by oxidizing agents or smoke condensate. Nakajima. K. tt d. (Tr.riY,1.) Tlre lorrnd o/ B1olotk.l Cliemlrtry 2S1(10) • 1027-1072,1979. From the School of Chemistry. Georgia Institute of Technobp. Atlanta; a.d the Merck Institute o( TDerapeulic Research, Rahway, N. J. HUMAN a•I-ANTICIIYMOTRYPSIN: INTERACTION WIiH CIIYMOTRYPSIN-LIKE PROTEINASES The data presented here wpport the previously suggested notion that *-I- antichymotrypsin (w•1-Achy) is a controlling proteis for the chytnotrypsi.-iika activity present in phagocytic eeda. The Mtteraetion of human plasma rl- Achy with serine proleiease from diaerent tissues was investigated. The pro- lein forns stable cornpleaes with pancreatic ehymwrypain, kukocyte eathcpw O, and mast cell chymotrypiin. There was rso inhibition of pancreatic trypsis or kukocyte elastase. With mixtures containing both rl-Achy and a-I-pro- teinase inhibitor, the former preferentially inac8vated kukocyte eathep.in U, while the latter had a strong preference (or pancreatic ehyrrsotrypsin. How- . ever, a-1-proteinase Inhibitor specificaBy inactivated leukocyte elast..e evev in 1:1 mixtures with chymotrypsin. The overall results indicate that oae of d,1-Achy's primary functions is to inactivale kukocyte cathepsln Q, whik a-1-proleinaae inhibitor controls the activity of other serine proteinases, notably leukocyte el.ata,e. TreriY, !., Bowen,l. and Sau6h, R. BlorhenrLtry 17:S6S1-S6S6, 197i. Other wpportr National Institutes of Ilealth. From the Department of Biochemistry. University of Geor6ia. Athens. 1 HUMAN a•1-ANTICItYMOTRYPSIN: PURIFICATION AND PROPERTIES ~ Human rl-antKhymolrypws (dr1-Achy) is a 6lycoprotein with two ur usual properties; it appears to be specilk for chymotrypiin-Irke enzymes and It is an acute phase protein whose plasma concentration may double within eight hours alier tissue damage. Consequently, this inhrbi:or may have a primary role in the regulation of specific chymotrypsin-like en:ymes, particularly those released during an inflammatory eplsode. Ihn report describes a rapid three- AII 51

step procedure for the isolation of o-1-Achy from whole plasma, and outlines wme of the inhibitor i properties. Ammonium sulfate fractionation, chroma- to2rapby on Cibacron Blue Sepharose at pH 7.0 and chromatography on SP- Sephadex C-30 at pH 3.3 isolated the inhibitor. About 45 mg of purified inhibitor was easily obtainable from 500 mL of plasma in less than a week. Its molecular weight was close to 65,000 and its carbohydrate content was approximately 26%. The inhibitor also was determined to have an amino- krminal arOnine and a carboxy-lermieal glycine. In add'ttio4 amino-termieal sequence analysis showed it to be somewhat honsologouw to aei .-1-proteioase inhibitor. Complexes of .-1-Aehy ..d butn.e ehymotrypsia or kukocyta eath- ep.is O are stable iri sodium dodapl wlfaM aod have molecular weipisu close lo 90.000, suggesting a molar 1:1 complex formation between inhibitor and enzyme. Tr.v/r, !., Oarner, D. and Bowes, 1. alocAemisrry 17:3617-3631, 1971. Other support: National Imtitufa of Heakb. From the Dep.rueeot of Biochemistry. University of Georgia. Athens. ENZYMATIC INACTIVA7ION OF HUMAN ALPHA-I-PROTEINASE INHIBITOR BY NEUTROPHtL MYELOPEROXIDASE The sensitivity of alph.-l-proteis..e (.-1-PI) toward chemical oxidanta has led 1o'a detailed iovestiptioa of potestial biobpkal oxidants that may be responsible (or this iohibilor's isacdvalion. Sucb a mccharrism would asuN in a lowered pulmonary capacity to inbibit the enzyme similar lo that found when there is a genetic abnormality. As is de+nomtraled in this report, acutrophil myelopsroaida.e can calalytiully i..c/1vaN .r1-Pl i. Ibe ptaeeee• of N,O, and chbride iort. The rate of the reaction Is directly proportional to both the myeloperoxidaas and .-1-PI otxroealratloa. and inversely proportionW to the H 0 concentration. As delermiasd by amino acid analysis of the oxidized arl-11, methionine is converted to the auttoaide form during the reaction and is the only amino acid that is modified. Of sipdficaoce Is the fad that this system does not affect either .-2-macroglobnlin or rl-aNichytnotrypsio, two other importael plasma proteiease inhibitors, nor cas il be replaced by horse- radish perosidase. It is possible that the myetoperoxidase released from kuko- cytes during phaltocytosis oxidizes the methionine at the art•P/ reaciive sile, thus inactivating this inbibitor and indirectly i«reasing the proteulyttc activity of these same cells. Matheson. W. R., Wooj, P. S. and Tr.rir, 1. aiocAem(cal and eloolYfskd Research Commt.nkations tl!<(2) :402 409, 1979. Other srpport: National Instilutes of Hesllb. 1`ram the Ikpartrncnt of PnxhemisAy. University of Geor6ia, Athens. I I , I I THE OXIDATIVE INACTIVATION OF HUMAN o-1-PROTEINASE INI11811OR. FURTHER EVIDENCE FOR METHIONINE AT THE REACTIVE CENTER The reactive center of human .-1-proteinw inhibitor (&-1-PI) bao previously been found to contain a methioei.e at position P,. Ilere, 1he im- porlance of this residue was tested by oxidizing Ihe inhibitor with increasing concentrations of N<hloroaueciniatide (SocNCI). There was  rapid lor of inhibilory activity toward trypsi., porcine pancreatk 14stase, and human leukocyte elaaase, correlating with the oxidalion of two (ou1 of a toial of eight) methionine residues to metldonine sulfoxide. Only moderate bues of cbymo- trypsia inhibitory activity were detected, however. Incubation of .rl-PI witb the four proteitases for extended periods of time resuhed in a necovery of much of the activity toward porcMe trypsin and human kukocptN elaalaae, but not toward porcine elaslase. The Nll,-/etmind sequence analysis of a larye peptide fragment isdated by papais dipestit>w of the oxidized .-1-PI confirmed that the reactive center tnelhionins had indeed been oxidized. Gel ekctro- phoresis demonstrated the slability of oxidised chymotrypsiu complexes; however. Irypsin and kukoc7rls elalde-rl-Pl cornpkxea were u.nabM, aad with both kukocyts aed pancreatic dastae, there was ep.pkte eonveraion of the oxidized Inhibilor to a lower molecular weight form. Together with the protective effect of melhionyl residues toward eyuw6en btvrnide cleavage con- ferred by the formation of Wive .-/-P1• trypsin compkxes, thae results strongly emphasize the intportance of protecti.p this iahibitor from chemical or biological oxidants. It is probable Mu ciprepe at.otce and other air pollutants eonlaia, or have the capacity lo forwr, oxidisiy agents which inaclivate a-1-PI through conversion of drc Pr arclhionine to tbe sulfoxide or sulfone, thereby reducing pulmoaary proteinase inhibitor activity. Sucb,a mechanism may pow significant in tbe p.thopeee.is of eo+pbysema. Johnson D. md Tr.vJr, !. TAe loMrwd o/ ebbtetc.f Cllantray 231(10) :1022-1026, 1979. OtA.r w/r)r.rtr Natio..11n1ihNa of HeaNb. From drt Department of Bioc6etaiMry. Utti.enity of Onorpia, Athens. THE CARBOXY TERMINAL SEQUENCE OF HUMAN ALPHA-1- PROTEINASE INHIBITOR In this atlemp to identify the mechanistn by which human alpba-/-pro- kinase inhibitor (.-1-PI) funcliorn, three Independent lechniaues were u.ed lo assay the inhibitor's carboay terminal residue. These Included digeslion with earboxypeptidases B and A, hydrarinolysis, and sequence determination of the carboxy terminal peptide obtained from cyanogen bromide fragmentation. Re- suits showed that when DFP-Ireated carboaypeptidase B alone or together wilh DFP•Ireated carbotypeptidae A was ulilixed, only ysine and a residue eluling under serine on the amino acid analyzer were released. Hydraxinolysia for 24 hrs., at s0' followed by amino acid anlysis also yielded tysine as ths earboxyl terminal residue of Signillcantly, the sequence of the single cyanogen bromide fragment obtained from .-1-PI which was devoid of homo- 32 53

4 i i scrine was found to be OLY-LYS-VAL-VAL-ASN-PRO-THR-0LY-LYS. This rosult con(irms that sequence also obtained for the carboay terminal o1 s-1-PI by S. K. Chan and unequivocally provides evidence that ly.ine is the carboay terminal of e-1-Pl. In adddion, carbosypeptidase C digestion iodicated sub- wantial degradation of .-1-PI by endopeptid.aes in the enzyme preparation. These roults suggest that free tryp.in, released from eompkzes, could readily activate the carboayl terminal lysisx of .-I-Pl, resultinll in osygen eschange with 11,0 in the medium. Travis. /. and lohnaon. D. elochemkal and siopAyskd Research ComwMwkarbnr a4()) :219-224, 1978. OOther arpperer National Institutes of Heahh. From the Department of Biochemislry. U.ivenity of Georgia. Athen.. STRUCTURAI. EVIDENCE FOR METNIONINE AT THE REACTIVE S1I1:OF HUMAN er-1-PROTEINASE 1NH/B(iOR Human a-1-proteiease inhibitor (ri-PI), the major serine endopeptidase inhibitor in plasma. plays an important regulatory role in preventing tissue protcolysis. Unlike other proteinase inhibiton, e-1-PI apparently functions by inhibiting serine proteinases of varying speeifkities at a single site. Recent attempts have been made to determine the amino acid sequence at the in- hibitory site of r1-PI, in particular that sequence preceding the reactive X- threonyl bond cleaved as part of eompkt dissociation. In this biochemical study, an unadecapeptide obtained by papain digestion of denatured .-1-PI was isdated and sequenced. l he structure of this peptide overlapped with the NII,-lermind sequence of modified inhibitor obtained from disso- eialed eompkaes of a-1-P1 with trypsin, ehymotrypsin, and elastase or frorn papain digestion of native .-1-PI, was found to be identical in all cases Further. more, structural homology with the reactive centers of proteinase inhibitors from other sourees was readily detectable. Significantly, methionine was found to occupy the apparent P, position in e-1-PI and the potential inactivation of the Inhibitor by osidalion of this critical residue could be imporlant in ob- taining  biochemical link with the development of lung disease. Johnson. D. and Travis. /. The lorrnal o/ Sioehrnrkal CAenrittry 23)(20) :7142-7144, 1978. Other supports National Heart and Lung Institute. From the Department of Biochcmiary. Uoivenity of Georgia. Athens. PURIFICATION OF HUMAN or2-MACROOLOBUI.IN BY CHROMA- T(XiRAPHY ON CIBACRON BLUE SEPIIAROSE A simple procedure for the isolation of e-2-macroglobulin (rorn hapto- 2lubin type 1-1 human plasma is described here. The technique invDlves the 54 I ! use of the multi-facetcd protein-separating abiliNies of Cibacron Blue Sepharose, which retarda lipoproteins and irnnwnollobtdins, the major contaminants in most .-2-macrotlobulin preparations, while separating the remaining plasma proteins according to their apparent molecular weights. Typical preparations yield highly active hornoeeenean inhibitor which represeMs appros3taately 40% of the .-2-macroglobulin present in the plasma sampk ehtomatoaraphed. A second step, gel flltration on UNrogd AcA 22, may be utilized to separate .-2-macrogk+bulin in contaminated fractions obtained after Cilsacroo Blue Sepharose chromatography. This method should greatly reduce any difficulty in obtaining sufficient quantities of this inhibitor for detailed structure and function sludies, and it could also facilitate studies of the properties of .-2- macrogfobulin isolated from individuals with various pathogenic conditiorr, such as cystic fibrosis. Virca, O. D., Trarl., l., HaB, P. K., and Roberts, R. C. An.fylkaf Qloc/k.niarry t9:274-271, 197R. Other aMpporf: U. S. Public Health Service. From the Department of Biochemistry. University of Oeorgia. Athens• aod Marshfkld Medical Fourdatioa for Medical Research and Educatio., Manh- Aeld, Wis. SENSITIVE SUBSTRATES FOR HUMAN LEUKOCYTE AND PORCINE PANCREATIC ELASTASE: A STUDY OF THE MERITS OF VARIOUS CHROMOPHORIC AND FLUOROGENIC LEAVING GROUPS IN ASSAYS FOR SERINE PROTEASES The development of sensitive aaays for human kukocyte and porci.. pancreatic elastasc involves esploitalion of the peptide sequenort MeO,Suc- Ala-Ala-Pro-VaI-X, a tetrapeptide sequeact dut is hiRhly specific for dasaw. In this study, variations in the leaving group. X, were examined to are how the sensitivity toward elastase detection could be improved. Thua, subslrates having X = -SBz1 (thioberwyl ester), -OEt, -AMC (4-methyl-7soumarylamide). or -NNapOMe (1-metholy-3-naplNhylamide) were synthesized and tested. The kinetic constants for the enzymatic hydrolysis as well as the sensitivities of all the substrates are reported here. Hydrolysis of the peptide .AMC aed -NNapOMe derivatives was followed by monitoring speelrofluoromerrically the release of II-AMC aod H-NNapOMe, respectively. Cleavage d/he shiobesuryl ester yields beeayl mereaptan as the hydrolysis praduct. MeO-Suc-Ala-Ala-Proa Val-SBtI, the best substrate of the series, was capable o/ detecting as littk as 2.4 pM (0.072 nee/ml) of activZ-sNe litrated human kukocyte elaslase and 5.2 pM (0.15 nR/ml) of .ctive-site litrated porcine pancreatic elaqaae uaing 4,4'dithiodipyridine. The corresponding values with F./lman: reagent were 3.0 pM and 7.4 pM, respectively. Advantages of this substrater are its high R,,,/K. values and ease of synthesis. lhe pepidyl-AMC is dmosl as sensitive as the thioben:yl ester and can detect 11 pM of H/L elastase and ifa pM a/ PP elaNase. An advantage of this substrate is the facl ,hat cleavage involves a peptide bond. llu peptidyl-NNapOMe has possible utility in histochemical 55

1 Y I a studiea. T he 4-nitroanilide assay enjoya moderate sensitivity and is estremely corivenient for routine use. Except (or the peptidyl ethyl ester aaay, all of the human kukocyte elastase assays reported in tbia paper are vastly more aeo- silive than any other one for this protease. Castilb. M. 1. et. at. (TrarL, !.) Anatrricaf slorikmi6trr 99:33-64, 1979. From the School of Chemistry, t;Teortia laatilute of Technolojy. A, laota; and the Merck Institute of Therapeutic Researeb, Rahway, N.1. UPTAKE ANO Rfi1.EASE OF CAL('IUM BY MICROSOMES OF NONVASCULAR SMOOIH MUSCI.E In previous studies. microsomal preparations from Rsrioea pig ileum Iontiludinal smooth muscle and rat uterus containing Mg-ATP (aa an energy source), and osalate anion (as a e.kium-trapping agent), continuously sequeslered calcium over a one-hour tirne period. The vascular muscle microsomal calcium uptake activity was Veatly stimulated by addition of dithiothredol, a potent deulfide reducing agent. to Ihe incubating microsonra. Addition of dilhwba-2-nitrohenroate, a sul(hydryl oxidizing ageol, depressed the calcium upuke. lhe present report extends this study to noavascular smooth muscle microsomes by esamining the distribulion of microsomal cal- cium uptake activoly on sucrose density tradients and looking at release of the sequestered microwmal calcium On sucrose density gradieots, calcium uptake of the smooth muscle micr.nomes appeared to be associated with intracellular membrane (sarcoplasmic reticuluml. Release of sequestered calcium from Ihe longitudinal muscle micrusomes was very slow (20% in 50 minutes). A small labile (raction (20%) was released by EGTA (I mM) in 10 minutes. Rapid release of sequestered calcium (909i in 10 minutes) occurred In the presence of Ihe calcium kxroplrure A27at7 (2 pM) or io the presence of chlorpromazine (I mM). Two other aRents, prostaglandin Es and p-eMoromeresuibentoate rrsod- estly enhanced the release of calcium. Chalurvedi. A. K., Fox. S., Sarrry, e. V. Rama. and Landon, E.1. Sioc/kmk.l end dbpAyrical Rrirurch Commrnkationr /9(7):1112-1119, 1979. OtAer support: National Institutes of Neallb and Biomedical Research Support. From the [kp.rtment of Pharmacology. Vanderbilt University School of Medicine. Nashville. Tenn. CIIOLINt?R('.1(' SYS7 EMS IN NON-NERVOUS TISSUFS Acclylcholine (A('h), which plays such an Imporlanl role in rhe cen- tral and periphtrnl nervuus systems, may also (1) play a rok in aon-nervow St. I I tissues and (2) have functions other than chemical Iransmission, even Is nervous tissue. In Ihe hope of ahedding some light on a posaibk common tola for A('h, information was eollecled on selected tissues without inoervNion and on tissues In which alternative functions for ACb had been poslulated -Tbs present extensive review examines this material and orRanitey It In1o such smlor sections as: Components of the cholinergic system; Oecurrenoa of one or mor. components of chdinergic systems In (1) orquisms and /iasues without tserves~ (2) tissues io which tbeir i.vdvcmcnt iw the nervous system is rernot.,, (3) tissues in which their invdvemeM is delineated into nervous asd soe-nervow components, and (4) cultured cells and developing eeR systems; Cbolinertic systems In (1) blood eeMs, (2) spernrdo:oa, (3) plaeenta, and (4) plaols; a.d, finally, the possible roka of ACh in non-nervous tissues. ACb occurs in several tissues where it serves so apparent neural furrction. At least in soare of tbcae Iissues. the ACh system is fully developed during maluration, growth, and pos.ibiy during regeneratio.. AC'Ir synlhesis does seem to decrease during aging in some ncrvous and non.nervoue tiuuea. While these observations should Pro- vide some clues for evaluating the role of ACh iw non-neural funclions, it may be more rewardinR to consider the role of ACh in non-nervoua tissues as similar lo that of a messenger. Sarrry. A. V. R. and SadavonSv'svad. C. rliarrrrarolotkaf Revkws 30(1) :65-1 32, 1979. OtAor nr'prtr U. S. Public Health Service and Vanderbilt Uaivenitry Research Counci. From the Department of PharmacoloRy, Vanderbilt University School of Medicine, Nashvilk, Te.e. .. PURIFlCATION OF SOLUBLE OUANYI.ATE CYCLASE FROM RAT LIVER The enzyme 6uanylale tyclar eatalyta /he formation of cyclic OMP ad is found in both soluble and particulate fractions of homogea.tes. Tbess tw forms difler in several respects including their antigenic properties. llrls tes Port describes the purification of soluble grranyla/e cyclase from rat liver and dctaNs some of its properties. On polyaerylamide,Rels, the purified enzyme displayed a single protein band coincident with i1s activity and indicative of a rnolecrolat weight of 110,000. When Mn /• was wad N the eolactor, 276.0 nnal of cyclic (lMP/mil/min were lormed, while 27.• nrrwl/mg/min were obtained with the use of M4t 4. These apparent specific activitin represent a 9,200-foid and a 7,400 fold purificNion, respectively. The activity of aohrbk guaeylale cyclase is not linearly related to Its concentration and the sne of increasingly higher enzyme concentrations tor assay at various slales of purification in- creases the apparent specilk activity of Ihe preparation. 1he purified enzyme is unstable at 4'(' and i% activated by nitroprnssikr nitric oaide, arachidona/e. Iinoleale, okate, and supero.ik diamu/ase. -Ihe degree of aclivaliNS. M.wever, is dependent upon she concentratirm of enzyme protein assayed. (3uanylate 17

cyclase appears to be a unirJue enzyme in that it can be altered by oaidative and reductive processes. The avaBaF>tlily o/ a highly puri0ed forns should kad lo additioea1 physicochemical aod kinetic studies of both the asaclive and active eniyme, and to the elucidation o/ the rnechaoism goveraio{i tedoa and free radical regulation of suanyfate cyclu. activity. Sraughkr, 1. M., Mittai, C. K. and 1Nrrad. F. proceedinp o/ rAe Nationaf Acadensy o( Sckwces oJ the United Srates o/ Awserka 76(1) :219-222, 1979. I Other anpp.rt: U. S. Public 1Hfsah! 3ervke, Diabetes Research Center and the Pharmaceutical Manufacturen Association Foundatioa. From the Division of Clinical Pharm.oolop, Deparlmests of Medicine aad Pharmacology. University of Virgiaia, Charlotlesvilk. I PROPERTIES AND REGULATION OF OUANYLATE CYCLASE AND SOME PROPOSED FUNCTIONS FOR CYCLIC GMP ~ ~ i Research studies of the cyclic OMP system have proliferated rapidly ro- cently, but to this date it is still not known how hormorses and perturbations of Ihe cell membrane precisely aller guanytate eyclase activNy, or what the functions of cyclic guanosine morsophosphate (GMP) are. In an attempt to confront these Qutslrons. Ihq review concentrates on some recent developments dealing with the properties and regulalion of guanylate eyclase. Sections of the paper are devoted to 11) the suhcellular activity and drsuriwtwn, properlies, actr.ation, and inhibdurs of 6usnrlare cyclau; (2) cydic GMP kveh in tissues; and O) smooth muscle rela.atrurs, secretion and Iransport, and a pouihk feedback regulator of tedos state and free radical formation a(unc- tions for cyclic GMP. From these studies it is becoming increasingly evident that guanylate cyclase activity and cyclic GMP accumulation are influenced by the redoa state of preparations, free radical formation in incubations, and fatty acid and lipid metabolism. While various hormorses can increue cyclic GMP synthesis in tissues, the precise events betweew borMorsc-receptor inter- action and guanylate cyclase activation remain unkrawn, representing one of the major puules in the fleld. Any or all of the processes mentioned above could serve the role of coupling hormone effects to guarsylale cyclase activation. There are other possibilities that also warrant consideration. While many hypotheses and clues have been pursued thus far, /he answers to these questions still remain elusive and some new approaches are needed. Murad. F. et 1. In: Greengard, P. and Robinson. O. A. (eda.): Advances In Cyclk Nwcleotide Research; New York: Raven Press, 1979, vol. 11, pp. 175-204. Other sr.pport: National Institutes of Health. f'mm the Orvalon ol ('Imicai Phsrmxolop, [kparlmenb of Medicine. Pharm- ..aloty, uk) Ane.rhtu.dory, llmversuy of Virginia, (-harlotlesvilk. THE OELATION OF ACTIN BY ACfIN-sINDING PROTEIN The fact that cytoplasmic eatracts can Iel has revived the speculatloa that tel-sol transformation might regulate cell structure and motility. Tbe soajor component of these loeb is filamenlous selia whkh, under appropriate coadl- tiau, forms a threedimensional aetwork, 7Tsis caa be achieved /n rirro by the addition of various purified proteins. Tbeoreticaqy, aay prWeia that has the ability to cross-link actia Alaments, such as myosie, can be employed. TTsir paper describes the use of two different arays to quantitatively compare aaia gelation by actin-biodiall proleia, myosin aed filamin: (1) direct AneasurenK.t of actin gel ri6idity, and (2) aedimeatabilNy, of P-actia in the preseea of jel-forming proteins. The sedimentation asuy was also used to aaser tha efed of proteolysis on the actin-gelliag activity of macrophate homornatea and lo esamine the role of variables that might control the actin and actia-biadiy protein interaction. According lo the data, the gelling action of actin-biadi.g protein is an order of magnitude more potent thaa filamin or myosin. When proleolyaft inhibitors were present during eell lysis and subsequent fractioaa- Iion procedures, 20% of the tecoverod actin-sedimenting activity in macro- pha6e estracts was isolated together with aetia-bindinR protein; in tbeir abseaa, lower molecular weight polypeptides with adia-sediarenting activity appeared and the specific actia-binding protein letia-sedirnerstirsg activity was red.eud. Other /adon, such as eowcesNratioa of MR++ or Cat F, temperalure a.d pH did not significantly aNer the equilibrisra e/fect of actin-biading protein oa actin sedimentation rates. RaisinR the concentration of KCI above 0.1 M d.r creased the sedimentation rate of actia in Iha presence of actia-biodialo protei.; trypsini:atioa of actia-binding protein for five wrinules abolished its actia-redi- enenting activity. In eorseMsion, aetin-bind'wy proteiw is a highly potent adia, cross-linkin6 agent: it is the major aetiw-gelling protein in rabbit luap es.ao- phages, and it is functionally distinct from the atructuallr related proteYs, W- min. srouchi, E. A., Ilartwig, l. H. and Stoaef. T. p. Thrlournal o/ fllo/osk.f CAerwisfry 2SI:t9t/-1199), 1978. Other w p port r U. S. Public Health Service. From the Medical Oncology Unit. Massachusetts General Hospitd, Depart- rnent of Medicine, l(arvard Medical School, Soston. OPTICALLY PURE (+)-NICOTINE FROM (i)-NICOTINH AND 81OLOGICAI. COMPARISONS WITII (-)-NICOTINE Central nervous system chbliner6ic receptors may be either nicotiaic or muscarinic. A growing body of evidence, nsoreover, suggests that 1M central effects of nicotine correlate better with its levels at nicotine receptors tham with its total concentration in different regions of the brain. Although so.ne reports dealing with nicotine binding and receptors have been published, not much has been written about the effects of optically pure (+)-nicotinc, the eaanti- omer of the naturally occurring (-) form. Tlse attempt here is to dnnotrtrate the pharmscoloeic stercospecificity of these nicotine iseNSSen. Optically pun SN 59

( I)-nicotine was obtained from ( 2)-nicotine by using a combination o( d tarlaric acid and di-p-toluoyl-l-tsrtaric acid. As demonstrated by their re- spective effects on blood pressure in anesthetized rats, on the isulted guinea pig ikum and on their lethality In mice, the di-d-tartrate salt of (})•nicotioe was kss potent than the di-)-tartrale salt of Ihe (- )-nicotine. -i hese results indicate that stereuspecrfirily has a substantial role in nicotine-receptor binding. Acero. M. A. rr al. lou.nd o/ Mrdkinul Chrmistry 22(2):174-177, 1979. Other aupprt: U. S. Public Health Service. From the Deparlment of Pharmaeology, Medical College of Virginia. Rich- nand. DI:TERMINATION OF HUMAN PANCREATIC CATIONIC 7 RYPSINOGEN IN SERUM BY RADIOIMMUNOASSAY Since pancreatic trypsin is capable of eQeeting a wide speelrum of physio- IMgical events involving peptide-bond cleavage in vir% many attempts have been made to detect pancreatic lrypsin in aerum by enzymatic metl..Ws. These were unsuccess/ul, however, and an alternative technique-radioimmunoassay -was developed for delectias of (mmunoreaetive forms of human pancreatic calionic Irypsin in serum. The trypsin employed as radioiodinaled Irxer in the assay was inactivated with tosyl-t.-lysine ehloromethyl ketone ('1CK) to prevent binding of the tracer to the serum inhibitors while maintaining its inmunoreactivity. lhe average normal serum level determined by this method was 26 ng/ml, with a range of 12-41 ng/ml. Eight of nine patients with acute pancreatic inAammation had at kal a 13-lold elevation of total serum immu- noreactive cationic Irypsin. The specifkity of Ihis assay (or immunoreaclive forms of pancreatic ealionie trypsin, as weN as the substantial elevation of serum radioimmunoassayabk Irypsit in acute pancreatilis, could make this a useful Mol for the diagnosis of difBcult easa of this disease. Among the mosl useful features of this radioirnmunoassay method are high specificity and sensitivity. Aho, importanlly, the asaay can detect inactive lorms of a pan- creatic prolease, such as the :ymolien, as weM u pro/ease-.,-antitrypsin com- pkses which might eaiu in plasma or serum. Close investigation of the nature of the radioimmunoauayabk calionic lrypsin in serum led to the conclusion that mosl, if not all, of this material is trypsinogen. This identification of cationic trypsinogen in blood implies that the :ymolien is secreted into the circulation by the pancreas rather than entering the bloodstream via ab- sorption from the intestine. GroAar. M. C. rr al. Arwrricon lornral o/ Physioluty 2)6(1):E77-Ee), 1979. O(6.r .upporr: Medreal Research Service of the Veterans Administrahon. N, w n. h 1 ah,.o.l..ry• Marrurcr Vetetanm AJrmmnIrahun d n«I rht Iwp.umtn/ ul Mtdount. (lu,rcr.uy ul ('.Ir. 1..••.•. `r"..r..r st..r., " ,r.... I . r EFFECT OF OXIDATION OF METHIONINE IN A PEPTIDE SUBSfRATE FOR HUMAN ELASTASES: A MODEL FOR INACTIVATION OF a,-PROTEASE INHIBITOR The reactive site of human .,-pro(ease inhibitor (.,-PI) contains a metr. ionine residue. It has been suggested that oxidation of this residue could dsr crease the prolease-e,-P1 reaction rate by reducing the enzyme: dfinily for the inhibitor. This report considers th4t e/lect of the methionine residue i oxi- dation by human pancreatic and leukocyle elastase on the rate o( hydrolysis of a synthetic tetrapeptide p-nilroanilide (SAAPM-NA) with a snethioniaa in the P1 position. Mild oaidatioa of this substrate yielda a methionine sulfoaide derivative, which is hydrolyzed by pancreatic elastas. 2 ud by ktukocyle elar tase at respective rates 3% and .79G of the onei measured when tAe pareM compound is used. According to thase resuNs, oxidation of the active aira methiunine residue of human .,.PI may decrease the inhibNor'a rate of rew tion with pancreatic or kukocyte eksstae. It appears, also, that the aateN so which oaidation o( the P1 methbnira residue in a,-PI destabilires protew binding depends upon the relative contribution o/ interactiona remole from the inhibitor i "active si1e." Del Mar, E. C., Brodrkk, l. W., Grokar, M. C., and LuRrnan. C. elochemkal and QioplYytic.l Rrarrch CoermunkaHona Et(2):246-)S0, 1979. Other asipporte Medical Research Service of the Veterans AdministrNioa From the Department of Medieine, Marlinez Veterans Administration Medied Center, Martinez, Cd., and the DepartmeN of Medicine. University of CaYY- /ornia School of Medicire, Davis. FORMATION OF A STABLE(-'OMPLEX BETWEEN HUMAN PROELASTASE 2 AND HUMAN a,-PROTBASE INHIBITOR The interaction of proel.stase 2 with plasma /w drro was studied 1. this attempt to understand the nNure of Me .,-protease inhibitor (P1)-aorud immunoreaclive elastase 2 that can be seas in normal plasam. Several Ynea of evidence are presented here which stroegly- support the conteetiow that proelaslase 2 reacts directly, in a relatively slow process, with .,-PI. Whe. rssl-labekd proelastase 2 was incubated with pooled normal plarna, two (rae- tions with' proelastase 2 binding activity wen separated by gel Altratio, on Bio-(hl A 0.5m. The minor [raelion was idenli/led as ..-macrodobuGa, whila the major /ractlon of proelastase 2, which eluled with the maior protein peak, was further purified by DEAEtellukose chromatography and by gel RhrNiow. ibis activity was found to purify wilh .,-Pl. A complex of proelasla.e 2 and o,-Pl, formed by incubating the two proteins for two hours at l7', was Mo- lated by DEAE-celhdose chromatography. Sodium dodecyl sulfate polyacryla mide gel ekclrophoresis indicated Ihat a 1:1 molar complex of M, = 77,000, which was unaffected by reduction and denaturalion by sodium dodecyl wl[ate, had been /ormed tncuhaliun of the isolsted complex with hydrosylamine, (rrl- bwed by gel filtration on Scphades (i• 1(10. dernonstrated that only proetssuse 2 was rekased front the complex. StM1arm dodecyl sulfate poiyacrylamide gel 61 M/

t ekctrophoreds of the fractions obtained by ilel 611ration indicated that the .,-PI rekased from the complex by hydroaylamine was reduced in molecular weight by approtimalely 7.000. When proetaslose 2 waa incubated for 16 hours at 23' with 0.1 M di'sopropyUharophosphate, 0.• moles of Inhibilor were incorporated per mok of :Ymogea The product no longer yielded elas- tase 2(dlowing incubation with bovine trypsis. The diisopropylphosphoryl- proelntae 2 was not abk to form a complex with .,-Pl, suggesting that the potential active site serine residue os the tytra6cn is roquired for complat formation with .,-PI. L.rpman. C., erodrick, 1. W., Ceol.r, 6t. C.. Si.cho, W. M., and lohnsoe, J. H. TAe lownd o/ dlolotk.iCliem6nl 231(17):af16-/323, 1979. Other awrpNt Medical Research Service of the Veterans Administration. From the Enzymology Research Laboratory, Martinez Veteruu Adminutration Medical Center. Marline:, Cal., and the Department of Internal Medicine, University of California School of Medkime, Davis. INFLUENCE OF CIGARETTE SMOKING ON DRUG METABOLISM IN MAN This report has two purposes: (1) to summarize the literature dealing with various aspects of cigarette smoke-induced changes in biolrans(ormalioe rate and (2) to review current research Ihsdinp regarding the ietera•tios of tobacco smoke ud Iheophrnine, a substance which haa been used to ssas the rok of smoking among multiple factors accounting for the great .ariabil- ' ily in biolrasatormatioa rates. Included is this p.per is a wmaary of drup whkh, when studied in rlre, clearly show either enhanced biwrnaformation or no difference in disposition in cigarette smokers. With the exception of ethanol, all the listed drup are metabolized by mictvsomd oxidative pathways. Numerous examples are presented of both an increased metabolic rate and absence of an effect of smoking on dru6 disposition is nun. This selectivity is consistent with the knoww ir.ductive eQects of P-41f stimulators in animal sys- tems. The lheopAypine studies show that both lobaeco and marijuau markedly alter ib ckara.ee in man. Ilowe.er, smoking is only one of swrnerow factors which can perturb hepatk biolransformstioa, r patient surveillance studies show lhat age as well as cardiac and liver diseass may be of greater importance in actual patients undergoing therapr. lr.4e. IY. l. D.rlp Aler.i.l4rw Rerf.ws9(2)•221-2)6, 1979. Other auppoNt National lnstituks of Health. From the Departmenl of Pharmaceulks. Sltle University of New York at SuAalo; l'/inical Pharmacokinelics (aboratory, Millard Fillmore Hospilal, Bulfalo 62 Ouerin, M. R., Stokely, 1. R., Hiains, C. E., and OrifRth, O. W. (Bd.)• Tobacco Smoke 1nAa/mion Sioorq CJiendslrp, O.t R/dpe Narbnd Le6o.eto7 Report 5424. 166 pp., October 1979. Natiowd Tecraical lefornutloo Sar.ioe, U. S. Department of Commerce, SpringfieW, Va. A report of research carried out by Oak Ridge Nalio.d Laboratoryr tor The Council for Tobacco Reseerch-US.A., lac., in colleboratios with Microbiological Associetes! setlksd., Marpland, and Process a.d lesdu- ments Corporation,s iroohysl, New York. This report details the more si6wiAcantt Mdiqa a.d analytical dr.ebpmeny of an appro:inutey three-year Inhalation bio.ray chpwistry research pro6ram sponsored by The Council for Tobacco Researeir--U.s.A., lac. The purpose of the program was to de0ne the characterislia of the Walton Horizontal Smokin6 Machine as an inhalNiow espo.ure device for twke. T1w study wr to be suAkiently detailed so b to wer Ihe Reneral concept of rodeM iehala- lion exposure to standing smohe, and it wae recognized that both monitoring concepts and analytical methods and 'auNrdmewtatiow would have to be devel- oped to do the task. The remaining abstracts in this seclion appeared in the above-t.amed pu6- lication on the pases irdicated. DESCRIPTION AND C1fARAC'TERIST'ICS OF THE WALTON IIORIZONTAL INHALATION EXPOSURE SMOKING MACHINE (1. R. Stokely'. l. H. Moaieyhwi, M. R. tltserins, L Flor.nN and 1. t7reewspaaa) (p. 1) An improved inhaluion exposure device, the Walton Horizontal Smoking Machiae, has been derised by the Prooen aAd ItrtrttsneMs Corporuion for exposure of smaR laboratory animals. The ebjeedw was to mechanically proa duee snake closely reptesentiK th.t produced by hwn.ws. Machine relwnn.ata and the evaluation of machine operation arrs described. Smoke of the proper, predicted dilution is provided; ks eomposNion agrees with that of smoke pro- luced by the Phipps and 11ird analytical smoking device except for having a slightly higher nicotine concentration. PROCESS AND INSTRUMENTS CORPORATION AUTOMATIC SMOKE EXPOSURE MACHINE-SEM 11 (1. It. Moneyhunr, l. R. Stoketyr anA L. Floranl') (p. 19) An nrtomalic Smoke Isposure Machine (SEM 11) has been designed and eonstrucled by Process and Instrytments Corporalion to generate a continuous stream of fresh cigarette smoke for smaN-rodenl inhalation experiments. Sane of Ihe problema a..ociated with intermilten/ amokMg systeem, aging of .mota. depk/ion of the aerosol by anirnah, and limitations of the numbcr of aniwnds simultaneously exposed are circumven/ed. The SEM 11 system ia described. its operating principles and charac/erislks are esplained and chemkal char- aclerizalion of the smoke delivered, as compared to an analytical system, is presented. 6J

ANIMAL CONTAINMENT VESSELS FOR NOSE-ONLY INHALATION BIOASSAY EXPOSURE TO TOBACCO SMOKE (M. R. Guerin', 1. E. Catont and W. E. Dalbey' )(p. 35) Whole body exposure compromises the results of inhalation bioassay test- ing because smoke constituents can be ingested by means additional to respira- tion. "Nose-only" exposure eiiminates these extraneous routes of administra- tion but the restraint required produces atrer which may inlluence dosimetry, longevity, and the resuits of the biowar. Several animal containment mecha- nisms have beee tested on hanwers, rab, or miee exposed to cigarette smoke from the Walto. Horizontal Smoking Mac6ine, the Maddo:/ORNI. smoke exposure system, or the Process and 1..lrumenb Smoke Exposure Machine. Retained dose depends on eontainmenl method. Slress from eonMement in restraining tubes was manifested by weight loss. Neck-only restraint has rov- erd advantages over previoutly used methods of containment. SENSOR FOR DETECTION OF TOBACCO SMOKE PARTICULATE MATTER IN INHAI.ATION EXPOSURE SYSTEMS (T. M. Gayk', C. E. Higgins' and 1. R. Stokely') (p. 47) A need for a simple, rapid, nondestruetive method of measuring the quantity of smoke in which animals are exposed has led to the development of a method based on light-scattering. A light<mitling diode and phototransis- tor array has been interfaced to both static and dynamic exposure systems. The diode ekment emits light into the aerosol, and light reflected by the smoke particles is detected by the photararoistor. Light-sensor readout is di- rectly proportional to smoke concentration and is independent of natural to- bacco smoke composition. The device is useful both for evaluating smoke inhalation systems and for documenting exposures. A CIGARETTE SMOKE MONITORING DEVICE FOR CONTINUOUS ANIMAL EXPOSURE SYSTEMS (T. M. Gayk', C. E. Higgins' and 1. R. Stokely') (p. 63) The safety of animals undergoing Inhalation e.posure 1o smoke requires that the snake concentration stays within acceptable limits. A nwnotor has been deveW+ped both for animal protection and for recording smoke concentra tion and exposure time. Total particulate matter is measured by a light sensor and carbon monoxide by non-dispenive infrared analysis. As used with the Process and lnstruments Corporationi Smoke Exposure Machine (SI:M 11), the monilor's signals activate an appropriate safety apparatus to proiect ani- mals from overespxrure. Results are prexnled from operational evaluation over several monlhs. I A METHOD FOR Tt1E SAMPLING AND DETERMINATION OF NICOTINE AND METHANE AS INDICATORS OF TOBACCO SMOKE CONCENTRATION IN INHALATION EXPOSURE SYSTEMS (L. B. Yeatts',1. 11. Moneyhun' and 1. R. Stokelys) (p. 73) Controling the amount of exposure to smoke that animals receive in as inhalation exposure system requires that the smoke concentration be accurately known. A general method has been developed so determine tobacco snaks coo- eentration in such a system by sampling and analyzing both Sas .ad particukte phases. Methane and nicotine, gas and particulate phase oomponeMa, respec- tively. serve u indicators of smoke concentration. S.mplinR smoke with a dual syringe, wilhdrawal-infusion pump and analyzing by gas chromatography per- mits determination of .nroke concentration on either an individual pull or compositeof-all-puRs basis. Results are more accurate whenn the composite sampk is used. Both indicators give smoke dilatiwu close to calculated dilutions. I ORGANIC GAS PHASE COMPOSITION OF CONTAINED SMOKE AEROSOLS USED FOR INIIALATION TESTING (C. E. Higpnai and 1. R. Stokely') (p. 85) The orpnie ps-phsa composition of smoke offered animah by the Wal- ton llorizootal Smoking Machine tor inhalation exposure is compared to that of smoke produced under standard conditions to determine the rekvawce of the exposure. Cryotherm.l trapping and high-resolution capillary eolum. =ss chromatography are used to investigate the effects of containment on the gas phase of cigarette smoke. There ae no signiAeaax diRerenees in smoke a6ed 5-25 sec in the Walton eaposure chamber. Mice reduce the concentratioa of gas phase eonstituents. with an indieation that high-boding constituents are rr- tained or otherwise in0uenced preferentially to low boikn. The orpnie ps- phase constituent composition of exposure chamber smoke is the same as tbat of standard whole smoke. However, constituents which exist both as gas phase aad particulate matter distribute to a greater degree into the gas phw M chamber smoke. DETERMINATION OF NITRIC OXIDE IN THE EXPOSURE CHAMBER OF THE WALTON IIORIZONTAI. SMOKING MAC/IINE (C. B. Ilonaker', A. 1). /lorton' and R. A. lenkins') (p. !7) Chromatographic separation M nitric oaide from almospheric oxygen 1w permitted the quantitative determination of NO in animal inhalation eaposure chambers. There appears to be little eiprette-to-ci6arette variation i. total PO delivery. Measurements inside the Walton exposure chamber without animals indicate that NO in 9% cigarette smoke hn a half-life of - 13 minutes. Ikcreases of NO n+ncentrations in the presence of animals suggest that the verate retaincd dose of NO may be as high as 7 roe NO per animal per cigarette. 64 65

I DETERMINATION OF PARTICLE SIZE IN TOBACCO SMOKE INHALATION EXPOSURE DEVICES USING METHYLCYANO- ACRYLATE! FIXATION AND SCANNING MICROSCOPY (R. W. Hdmberg') (p. 103) A method for measuring particle sise distributions in cigarette tobacco smoke is discussed. Smoke particles an Rsed in the gas phase by cont.ctinp them with methyl-2cyanoacrylate vapors ead are collected on Nuckpore dlten for scanning ekctron microscope (S8M) sptaetin.tion. Diameters are dcter- mined from SEM photographe using a graphic digitizer. Distributioe cbarae- terislica ar* cakulated from stored di.aaMr values using miokomputer-bued ptorrams. A simple porlabk sampling technique suitable for use vnder bioassay e:- posure conditions is described and esamined for possible bias. Size distribu- tiona typical of esposure corditions are reported. A ME771OD FOR THE DETERMINATION OP TOBACCO SMOKE INHALATION DOSIMETRY USING CARBON-14 LABELED DOfR/ACUNTANE (1. E. Cato., Jr.t) (p. 119) A facility is established to provide analytical support of investiptions into the influence of exposure conditions oo tobacco smoke inhalatioe dosi- metry using mice. A method (or more rapid sod accurate analyses of tisues for carbon-14 and for processing the large quantity of data generated was needed to meet support requirements. Rigorous cootrols were included in the dosimetric method to ensure reliability and lo allow eomparisons between exposure parameters. INFLUENCE OF SMOKINO CONFIGURATION ON PARTICULATE DELIVERY (1. R. Stokely$, M. R. Gueria' andC. K. Bayoe') (p. 1) 1) A single poet smoking machine and the W.Itoo Horizontal Smoiing Ma- chiae were wed to evaluate the iaAue.ce of sr0okiag conBguratioa oo deliver- ies of tar .ad nicotine. Results ue eompered with standard deliveries u deler- mined winp a Phipps and Bird analytical smoking machine. The cigarettes dclivered the same quantities of tar and nicotine is eilhet a vertical or hori- wntd position. Free smoking results is an Increased delivery of Iar and, e.pr ciady, akoline as compared to reatrictive smoking. The Walton Hori:ontd Smoking Machine produces the same delivery of tar and nicotine whether it is operated u a reverse smoker or the smoking mechanism ia modifkd to provide draw pu1Rng. CARBON MONOXIDE AND CARBON DIOXIDE CONCENTRATIONS IN SMOKE AS A FUNCTION OF INHALATION EXPOSURE USING MICE (I. R. Stokely'. M. R. Ouerinr and D. D. PaK') (p. 137) The concenlrations of carbon mono:ide aad carbon dioxide in tlx inhala- tiwn chamber of the Walton Horirontal Smoking Machine while e.posing 20 ('37 black mice were determined as a(unction of esposure history. ('arbon monuaale concentraltons were in the espected range, and eatbon dieside con- 66 tributions to the inhalatioa atmosphere by animal respiration did srot exceed 0.5% (v/v). Esperieaced animals each depk/ed appro.imakly 1% of the available carbon monoside frare a 10% (v/v) smoke from /he Kentucky 2A1 cigarette. Carbon dioaide evolution monnitori.R wgge.te that the mia quickly learn to avoid smoke inhalation. The avoidance meehanira «aehea Ila f4ak afler two weeks of smoke exposure. MONITORING HYDROGEN AS A MEASURE OP'SMOKB CONCENTRATION (T. M. Oaykr, C. I? Higifiru~, J. H. Mo.eylw.' and 1. R. Slokelr') (p. 149) Thermal conductivity of tbe gas phase of tobacco smoke may be nwd- tored to provide a eoatinuous measun of seake coacentratio.. lLe rnpoass is due almost solely to the presence of hydrogen whew the nw.itor is Attea with a water and carbon dioaide acrubber system. The device is rse(ul r a naeilor of rnoke concentration in inhalation exposure syNema. V 1. InsrwYno[o" and Ad.ptire Mee6wnlaaxa COMPLEMENT (C3)-ACiIVATED PHAOOCYTOSIS BY LUNG MACROPHAGES The interaction of rabbit lu.ll eucropia6es with EarAerkAb cob Ypopoly- saccharide eoated oil dropkta opsonized with a(raBmesl of Cl, a.d witr albwtwneoded oil dropiets opsoaited with aMialbwnin IRO wr invaliplea. Detailed eaamirntion of C)-dependent. IRO-aetivated, and nonspecific phyo. cytosis by Irypainined rabbit lung macrophaBts showed that the C3- a.d 16Ci-mediated ingestioa by pulmonary .nerophatia have different kinetic pat- terns. Trypsiniz.tion of macrophapes and removal of divaknt eatior from the medium primarily aNer the kineties of C-mediaKd ingestion, dfecti.g r initial rapid ingestion phase that is not evident during* 1gO-medided Ingealio.. The eRects of trypsin were reversibk i( the macrophalles were Incubated M proteinconlainins mediurn. Inhibition of maeropha ge proteie synthesis wilh cyc/oAeximide prevented the reversal. The e.perimemal results ptesenled bere support the notion that the functional di0erences between these two typse of ingestion mechanisma are mediated by distind membrane .nolecuke or do- mains. It is speculated that the C) "reeeptor" of lung m.crophalies is a func- tional eompka with al least two componenta: a divalent eation-depeedeM, trypun-sensitive fundion, and a divalent eation•independent trypain•rcsisuM function, which more nearly resembks the 1tO-activated Ingestion mechanism. . Shurin, S. B. and Sroiuf. T. P. The /orrrnd o/ hnerrwoforr 120(4):1)05-1712, 1979. OOther aupportr U. S. Public Heahh Service. From the Medical OncoMgy Unit, Massaehusetts General Hospilal, Dep.rt- ment of Medicine, Harvard Medical School, Boston. 67

4 .1.. 0 ! L~ ~ m 4.11 ~ B m IMMUNOCHEMICAL AND PHYSICOCHEMICAL PROPERTIES OF TOBACCU E7CTRACiS In this immunological study, estracts of green tobacco kaf, cured tobacco kaf, and cigarette smoke condensate were prepared and certain of the physico- chemical and immunochemical properties of theae materials were investigated. Analyses by isoekctric focusing revealed as maey as 20 bands with green kd extract; with cured leaf extract and smoke ooodeasate extract there was a marked decrease In band number. The pNlern for smoke coodemau: extract was similar to that (or cured leaf eatrad. Ittsnwntizatioo of rabbits and guinea pigs with green ka( extract yielded aalsera that produced at least (0 bands with this material; these antisera reacted weakly with cured kaf extract .nd not at all with smoke condensate extract. Astisera to cured kai extract re- acted weakly with both cured and gree. kaf extracts. None of the antisera formed precipilin bands with smoke eondenaate extract. Serum apecim:ns from sia rabbits Immunized three times with one mp of smoke eondensalo extract in Freund's complete adjuvanl at monthly intervah, and reimmunized eight months later by intravenous injection of smoke cordetwte extract Ixoduced precipitin bands when reacted with Sreen kai and cured leaf eatracta. After isoekctric focusing of green leaf estract, imnwno/laation analysis using rabbit anti•Creen leaf extract showed that the antigens were acidic proleim with iso- ekclric points of 4 to S. These results indicate that aeveral antigenie moieties are present in tobacco leat; they also point to differences in the anlirenicity of the various tobacco cslracts. GfrirA. G. l. and Welsh. P. W. Arnerken Rer/ew o/ Rrrpirarory fln.arr t20(S):995•1001, 1979. Other support: Nalwnal InrtqWe of Alkrp and Infectious Diseases and the Mayo Foundation From the Departments of Medicine and Immunology. Mayo Foundation. Rochestcr, Minn. IMMUNORFACTIVE FORMS OF HUMAN PANCREATIC C1IYMOTRYPSIN IN NORMAL PLASMA This methodological paper rrports on a radioimmunoasaay for chymo- trypsin 11 that has been used to measure totd imtswooreactive humam pan- creatic chymotrypsin ia normal plasma and to characterize the molecular forms of this prolease in the circulation. For this naay, phenylmethanesulfonyl fluoride was employed to inactivate the chymotryptin 11 used as radioactive tracer, in order 1o prevent tracer binding to the plasma proteau inhibitors while maintaining imemnoreactivity. Pancreatic chymotrypsin I and both zymoffem, ehymolrypsinopns I and 11, cron-reacted strongly in the assay. However, no other human pancreatic protease cross-reacted to a measurable extent. When the molecular forms of immunoreauive chymotrypsin in normal plasma were investigated by gel filtration on llio•Gel A-0.Sm, two peaks of immomoreactive chymotrypsin of approsimalely equal size were observed The first peak ,rheted in a position cr+nsistent with an a,-anlilrypsin:ehymotrypsin annpleR aful the v.aud eluteJ Muh a nwleeular weitht of approsmtately 68 t 25.000, corresponding to the elution position of chymotrypsin. In summary, this radioimmunoauay for chymotrypsin 11 has been employed to detect an of the immunoreactive forms of human pancreatic chyrnotrypiin, including chymotrypsin I and both chymotrypsirwgic tts. Chymolrypain I and cbymotrypM sinogen 1give slightly nonparallel dose-response curves in the radioimmuso` assay system. However, since both proteim eron-react strongly in this away system, it has been possible to obtain data eoncernittp all of the molecular forms of immunoreactive human pancreatic chymotrypain in plasma. ;ro4ar, M. C. er d. The lorrnd o/ eloloticcd Cbemtstry 2S4(t) :2775, 2781. 1979. Other surprt: Medical Research Servioe of tha Veterans Administration. From the Enzymology Research Laboratory, Martinez Veteram Hospital. Mu- tinez, Cal.; and the Department o( Internal Medicine, University of California School of Medicine, Davis. SUPPRF.SSIVE EFFECT OF ALVEOLAR MACROPIIAt3ES ON THE IN VITRO IMMUNE RESPONSE OF RABBIT LYMPHOCYTES The role of macrophaps as regulatory eelh in the immune resportas hr become an intense area of research. Numerous reports have described atrp- pressive effects of macrophaRes that are mediated by celleell contact or throud the elaboration of soluble factors. This paper, which represents the flnt report on alveolar macrophage (AM) funclion in humoral immune respomes, de- Kribes a suppressive effect of the AM in the Reaeration of the plaqse-forr.i.l cell (PFC) response in rabbit lymphocyte cultures and provides evidence for the involvement of a soluble faclor(s). At a ratio of I AM:10 lymphoid eelY, the PFC response was reduced by 90%. At rMios of I AM:1,000 ee1M or higher. Ihe responses were not signifkanlly affected. Addition of lymph awdr. thyntus or spleen rxlb at similar ratios did not cause suppression of ths PPC resporne, thus indicating that neither ceq crowding nor depletion of wtriew was a plausible explanation of the suppressive effect. Separation ot apken ee1N from the AM by a semipermeable membrane also resulted in suppresaion sad urMested that soluble regulatory (actor(s) were involved. This supprcssion required the presence of AM during the /lra 24 hn. of the culture period; addition at 411 hn. resulted in only partial suppression and addition at 72 hn. did not have any effect at all. 'I hese and other results presented in this paper and elsewhere suggest that AM may play a repdalory rok in imnlune re- spnnses. 'Ihe importance for such a role In rivo and characteriration of the suppressor factor are being investigated now. Pennline. K. l., Conrad, R. E.. Oerber, 11. R. and //ersrowltz. H. e. Journal of the RrHcrloendorhrllal Sockry 2S(3):493-312, 1979. Orher.upport: National Institutes of Ilealth. From the Department of Microbiology. Georgetown UnivenNy Schools of Medicine and [kntntry. Washin`ton, 1),C. 69

0 GENETICS OF XENOfROPIC VIRUS EXPRESSION IN MICE: 1. EVIDENCE FOR A SINGLE I.OCUS REGULATING SPONTANEOUS PRODUCTION OF INFE['f1OUS VIRUS IN CROSSES INVOLVING Ni.B/BINI AN[) 129/1 STRAINS OF MICE In this attempt to study the pattern of segregation of infectious X-tropic virus expression in homogenized spknie tiasue. genetic crosses were made be- tween high-virustspressing NZB/BINI mice ud non-virussspres.ing 129/1 ones. The results presented here show that spontaneous production and release of infectioaa X-tropic virus in crosses between the two strains segregate as a singk, autosomal dominant-like gene. Analysis of Infectious virus produ:tioa in accond•ba-kcross families IF, X 129) x 1291 eon6rmed this conclusion. Variations in the mounl of X-tropie vinr released were evident in all genetic crosses. Virus titen (e.pressed as focua-formin8 units per milliliter) of super- natant fluid ranged from high levels in the NZB mice to somewhat lower levels In crosses involvinj the 129 mice. Since all mouse strains seem to contain the same number (sia to nine) of proviral copies of X-tropic viruses, the data from these tenctie studies suggest that this gene may be regulatory rather than structural. It seems possibk that derivation of eongenic lines may prove useful in delineating the nadne of these gene functions and provide a system relatively free of rnodrlying influence so that the tok of X-tropic viruses in such diseases as autoimmunity and cancer can he addressed. Ltvy, 1. A., Joyner, /., Nsyar, K. T. and Kouri. R. E. (Microbiological Aao- riun, Inc.) Journal o/ Yiroloty )0( )):734-73a, 1979. • Ot6er.upportt National Cancer Institute. From the Cancer Research lnstitute, Department of Mcdicine. University of California. San Francrsco; and the Department of Biochemical Oncology. Microbiological Associates. Inc.. Bethesda, Md. EFFFCfS OF CIGARETTE SMOKE ON ELASTASE SECRETION BY MURINF. MACROPHAGES Since alveolar macrophages had been shown to secrete elastase in vitro, it seemed worthwhile to study the effects of cigarette smokioS upon elaslase secretion by murine macrophaffes. Also studied bere, were the effects upon elastase secretion of adding cigarette smoke in vitro to cultured macrophages. In the in vivo eaperiments, mice were chronically esposed to cigarette smoke for various time periods up to four weeks. As a consequence of the e.posure, there was an increase in the mrmber of alveolar macrophaaes obtained from the lungs of these mice, and examination of these cells by light microscopy revealed increased numbers of highly pkomorphie cells f111ed with pigmented residucq of cigarette smoke llx exposed cells showed more mitotic activity (with a Rvclodd increase in the number of alveolar colony-forming eells) than the 400111411% When m.aasph..ges drrrved irum mice e.ppned to cigarette smoke ..rIc .uhnr..1 at h.sh J4 o.dv rn the .Lsrnse cd serum. Ihey sectetcd sogrufiramly Rf..1., .n.......r. ../ r/....v rh.n .1.1 Ihr unw nmio/hrr of s4rNrul ntacu.phalrs 1/.luh, ......1. .r... .1 i. .../..1•.~~.1 tl.o..r o.', IfINon ll.-m IwNh Ihc aunlrnl and exposed cultures, even at very low concentrations. These studies showed that chronic cigarette smoke inhalation by mice led to heightened mitotic activity of alveolar macrophages, increased numbers of macrophars in broecho- polmonary lavage fluids and doubling of the rate of elastase secretion by these cells when cultured in vitro. Several possible mechanisms are considered bere to account for these results. White, R.. Whi1e, 1. and Janos, A. Jorrnd o/ Laboratory and Clinical Alydklnr 94()):4t9-499, 1979. Other arpporat National Heart, Lung and Blood Institute. From the Department of PathobSy, State University of New York at Sto.y Brook, Stony Br«sk. EFFECT OF CK3ARET7 E SMOKE EXPOSURE ON MATURATION OF THE ANTIBODY RESPONSE IN SPLEENS OP NEWBORN MICE This study attempts to rktermine if exposure of neonatal animab to smoke has any eRcct on the maturation of inwnune r~spa~arveneaa d nseasured by tly appearance of splenic plaque-forminR cells (PFCs). Wilhin their ArN 24 hows ant. until some were 10 weeks of ap. Balb/e mice were exposed for various periods of time to the smoke of Kentucky Reference 1 R 1 cigarettes in . Prototype Mark 11 Walton Hori:oMal Smoke Enposure Machine. For the /lrst nine days, there was no difference in the magnitude of PFC response be- tween the smoke-exposed mice and the untreated control animals immueited with sheep red blood cellt (SRBC). BeRinninR on the 101h day, however, .ed each aubsequent Iime, the mice exposed /o semke showed a statisticdly aig. nilkant reduction in the rwmber of apkoie PFCs: 33% on day 10, 60% on day 14, 90% after 4-10 weeks. Such marked inhibilion of the mawratiom of the spknie aMibody response to SRBC is consistent with previous reporM of the innrwasuppressive effects of cigarette smoke exposure in experimental animals and in humana. It ia possible that this irrhibilory effect is manifest at the level of the macrophatie or thymus-derived lymphocyte in the spleen. Both types of cells are thought to be functionally inaeature in the newborn mouse Less likely, but still possibk, is the notion that this effect also operatcs at the level of the B cell. I Hrrttowirt, H. s. and Cooper, R. B. ftdiatrk Research 1):9l7-991, 1979. From the Department oF Microbiology. Georgetown University Schools of Medicine and Ikntistry, Washington, D.C. . IMMUNE ACTIVATION BY T-INDEPENDENT ANTIGENS: LACK OF EFFECT OF MA('ROPIIA(:E DI:PLEttON ON TIIE IMMUNE RESPONSE TO 1 NP-LPS. PVP AND DEXTRAN While it is well known that nsscrophates are required for immurre resp/nses to Ihymus•dcpendcnt anligens ~uch as sheep red Mood cells (SRBC), the involvement of macrophages in immune respoma to thymus-independent 71

antikens is less ckar and somewhat controversial. In an attempt to understand the requirements for macrophages in these last reactans, carrageenan and rabbit antimacrophage serum (AMS) were used to inhibit macrophage func- t'an in BALB/c mice and to deplete macrophages from spleen cell cultures. When large doses of carrageenan (e.g. 10 mil) were injected intrapetitoneally into BALB/c mice in order 1o inhibit macrophage activity both in the pcritoneal cavity (the site of antigen deposition) and in the spleen (the site of antibody tormation), the immune resporw to SRBC was markedly reduced at all Immunogenic doses tesled. Howe.er, the responses to E. roil LPS, polyvinyl pyrrolidone. and deatran B-13335 were unaffected by such treatment, suggesting thal severe depletion of eueropbaRes in rivo did not affect the response to these thymus-independent antigena. The In rirro immune response to 1 NP-LPS was also unaffected by either earralleenan or AMS treatment. The results of tM !n viro and in rirro eaperitnents which utiiired carrallrenan and AMS sugj!st that the immune response to thymus-independent antigens does not require the participation of macrophates. Wong. D. M. and Nrrrcowltt. N. 1. Immwnofo#y )7:765-773, 1979. Other support: U. S. Public Health Service. From the Department of Microbiology. Georgetown University Schools of Medicine and Dentistry. Washington. D.C. BOMBESIN-LIKE IMMUNOREACTIVITY IN THE AVIAN OUT AND ITS LOCAI.IZATION TO A D1SIINCT CELL TYPE Radioimmunoassay and immunocytochemislry were combined here to study the distribution of a bombesin-like immunoreaclivity in the avian lastro- Intestinal tract. The radioimmunoassay of tiasue ealracts showed that the largest quantities of this material were present In the proventricuhn with smaller, but still eonsiderabk, amounts in the gizzard. Immunocytochemicany, the e.tract- abk bombesin-like immunoreactivity was localized in numerous endocrine cells. Electron micro.copy revealed that these eelb contained small election-dense secretory granules of a type not fully characleriud in any previous ultra- atructural classifications. lhus. the use ot the semi-thin/thin approach has revealed the presence of a homhesin•like materid in the chick gu1. Tl+e name BN is proposed for this cell type in accordance with Ihe principle of clas- aifyins endocrine cells by this funclion. Timson, C. M. er. at. (Will, I. A.) Nistoc0em6try 61:21)-221, 1979. Other support: llx Medical Research Council and Farmitalia. From the Departments of Ilislochemislry and Medicine, Royal Postaradnale Medical Schonl. ILmmcrsmNh Hospi/al. l.orwlon; ('enaro Diagno.uca /+to- patolrrgica, Univer+rti dr Pavia a Varese- Varese, Italy; Istituto di Anstomia e IsNda`u PataloRa a dcll' Universitl di Pavia. Italy; and l he Salk Instinde, PcphJe IIr4rlo2y 1 alr.ralory. San 11iego, ('al. ; i TIIYMIC MACROPHAGES MODULATE ONE STAGE OF T CELL DIFFERENTIATION IN VITRO Thymic macrophages may play a physiclo=ie role in the regulation of at kut one stage of Ihymocyte maturation. Evidenoe supporting this hypotheais is presented here through the evaluation of the rok of isolateo thymic macros pha6es in Ihe stimulatioe of thymocyle maturation !n .1tro. The prooedun for isolating thymic mouopha6ea is described. Culturing Rradient-puri6ed immature thymocytea on thymic macrophaRes increasa the eaprenion of mouse major histocompatibility cornplea antigen and decreases lytic sesl- livity with aruithymic kukemia antigert and oompkmerN. The macrophake- aimulded thymocyle can aho respond in the mixed lymphocyte rsactioe. It is suggested that the macrophaRe may regulate one stage of lhynsie diAerentia- /ion /n riro. Beller. D. 1. and Uw«rw, E. R. The lorns.f o/ lmwrrwolory 121(3):1661-1t61, 1972. OtJier wpports Natiopal Institutes of Heahh. From the Department of Pathotop, Harvard Medieat School, Boston. IN VITRO SUPPRESSION OF SERUM ELASTASE-INHIBITORY CAPACITY BY REACTIVE OXYGEN SPECIES GENERATED BY PHAGOCYTOSINO POLYMORPHONUCLBAR LEUKOCY7E4 This shdy demonstrates that human polymorphomrckar kukoeyta (PMN) stimulated by eaposwe to opsoniaed anligen-antibody compkaaa rekase dialytabk oxidant species, which in turn are capable of inacti.atMR the elastase-inhibitinR capacity (EIC) of whole human serwn or of puriAed human .,-proleinase Inhibitor (.,-Pi). Superoaide d'anwtase, catalass, or mannitol partially protected EIC, suggesting that hydroxyl radical, formed by the iaMeraction of auperoaide radical and hydrogen peroxide, may be respon- sibk for this effect. The tact that EIC was also partially protected by N.Ns implia that myeloperozidase-mediated reactions are involved as wep. A aanthine and aanthineoaidase mialure, utilized as an artificial superoaids radieal-fienerating system, could be subslituted for phagorytosing PMN with resultant FIC suppression. These resules are consistent with those of previorr studies which demonstrate the release of poleaA oaidants by stimulated PMN, and with other data that establish the sensitivity of a,-Pi to inacNvatioa by oaidants. The oaidatiw inacliwtion of proleinase inhibitors in the micrs envirotunenl of PMN that accumutats at inAammation siles may allow pro- lenes released from these celb to damage the adjacent connective tiaus ~ strrktures more readily. Carp, H. and lenoff, A. The lorrnal o/ t7inic.f Inrrstitadon 6):793-797, 1979. Other srpportr National Ileart, t.ung and Blood Institute. From the Department of Pathobgy, Hedth Sciences Center, State University of New York at Stony Brook, Stony Brook. 72 71

SUBSTANCE P-LIKt31MMUNOREACTIVE NERVES IN MAMMALIAN LUNG Although the autoeomic nervous system has been classically divided Into +drenegic and cholinergie nerves, a third eoaeponeot, P-type (peptidergic) or purinertic nerres, has now been ideatibed oa the basis of uhrutructural obser- vatioes. In the present study, specimea trom all regions of The lung were obtained from 21 adult rats, t guiom pip, 4 tsioe and 6 neonatal pigs of both seaes and used for indirect immuwiworesoe.ea, Results showed that fine vari- e:ose fiben, displaying substance P-M'ts iw.Maoreactivily, were found in the lungs of ap animals studied usd wera simitar in appearance to those seen ie the control sections of colon. The .erve fibers displaying substance P-like immuno- reactivity were observed is association with blood vesseb and airways of the lung. Immunoreactive Aben were observed in a.d around bronchi, bronchioles and tnors distal airways. In general, this inwmt.ocytoclsemical study provides tnorp(sobkical evidence for the Prwwo. in tha lung of att eatensive system of nerves containing substance P-1ie Imwwsoreaetivity. Then has already been increMinb evidence that the auaramie iMenraion of several organs coMaase a peptidergic component and it would now appear that the hrsg is yet another orpn which is innervated in Ihis manner. Substance P is known to have a potent vasoddatory elfect which cannot be blocked by adrenergic or cholinertie blocking aCents. Thus, In addition to a possible sensory role in the lung, these substance P-like immunoreactive fibers which are distributed around blood vessels and airways may /orm part of a complex system regulating lung fune- tioo. Wharton, 1., roloR, I. M., Bioom, S. R., Wifl, I. A., Brown, M. R. and Pearse, A. Q. Inreitigatire Cell Pathology 2: )-10, 1979. OtAer suppsrtt Medical Research Council. From the Departments of Histochemistry and Medicine, Royal Postgraduate Medical School, Hammersmith Hospital. London. MACROPHAQE-TCHLL INTERACTIONS INVOLVING LISTER{'A MONOCYTOGENES - ROLE OF TItE 11-2 GENE COMPLEX Resistance to lJsreria Infection requires both generatioe of immune T cells and activation of macrophatZes for enhanced microbioeidal activity. From studies in the mouse, it would appear that the Interactions between lymplw- eyles and macrophagcs are probably regulated by genes in the major histo- compatibility gene complex (MHC) of the species, especially The murine 11-2 complex. The studies reported here were undertaken to determine the manner by which the T ceNs inleracl with mactophages and t.liter(a, 1he kinds of elfects taking place aher such interactioru, and the steps under ditect cunt.ol of the MIIC. lhis interaction was assayed either by determining the anuwmt of thymocyle miaoIcn in cultute fluids after 24 houn, or by estimatinl the I 0 degree of T cell proliferation after 96 hours- Each assay depended on critical concentrations of macrophages„ T celb, and heat-kilkd I.lueria; both assays required specific Llueria-fumeune T eella. For an eQective iateractioa, 1ha T cells and the macrophagea had to share the I-A region of the H-2 BrM eompkx. Macrophages bearing la, which represented a minor population of macrophages, were essential for the proliferative response to encrophake- associaled 1lsteri.. Aho, la-bearing macrophages were an importa.t coasw ponrnt in the interactiona leading So increased secretion of mitoges. Fi.aUy, the immunogeeaic anoiety associated with Listeda was ahortJived, disappearing 24 houn after uptake of llireria by maerophqea. Fart, A. O., Kiely,l-M. and Un.ww, E. R. The lorrnd o/ Immunology 122(6) :2195-2404, 1979. Other arpp.rrt National InaNuteaof Health. Prom the Department of Patholopr, Harvard Medicat School. Boston. c INDUCTION OF CYTOG9DAL MACROPHAQES AFTER IN VITRO INTERACTIONS BETWEEN USTERIA-IMMUNE T CELLS AND MACROPHAGES - ROLE OF H-2 As part of a continuing study on Ihe interaction of macroplulles ad T lymphocytes from mice inrrwnized with wrer(s nsonocrrotear., two tAect. that became apparent drrine culture of these T cells with snacrophaBea having ingested The organism were aludiedd eoth tha secretion of Irmpho.tir.ulNory, amolecules and T ceNa proliferation increased. Culturing teacrophagea from nonimmune mice with T cells from mice immune to LJite.le for 24 hours Induced eytocidal aetiritr in the maerophqes, as demonstrated in a cMomiuwr- releaae assay with PMtS tumor eeRs. The culture /luids eontained a mokcule that induoed this cytosie activity and tbe irMCrad'aws leading so it required an inductive event is which Me major hitloewnpatibility complex had an essential role. This process was found b be restricted by the 11-2 Ber oaw- pkx. AppareMly, The activating factor acted upon aynbeneic or allogateic macrophabes, or those /acking detectabk la molecules. '11m imemrsobenic mwieqr associated with Llrrerla was short-lived. Several conditions were established for the generation of eylolytie macrophages: (1) specitkity of T cells to t/iteria and the need for tnaerophages to specifkally interact with these heN- kilkd organisms; (2) homology at the I region between macrophatoes and T. celh, and (3) the presence of macrophqes bearing la molecules. Farr, A. O., Weehter, W. J.. Kiety, l. M. and Unanwe, E. R. The lorond o/ Immrnobp 122(6) :2405-2412. 1979. Other arpportt National Cancer Institute and National Institutes of Heahh. From the Department of Palhology, Itarvard Medical School, Boston. 74 1 .75

. AUTOMATED RADIOIMMUNOASSAY OF NICOTINE An automated radioimnsunoaaay of nicotine in a nooequilibrium system has been developed for the estimation of nicotine In biological fluids. In this paper, a eicotioe antibody is characterized and its use in Ihe automated radioimmunonsay is described. The u.e of a rad'aiodinued (rssa) nicotine derivative as a tracer In this system pve a sensitivity of a 10 K/1 for nico- tine with a between-run precision of 7.4% and wilhin-rwt precision of 6.0%. In experimental triab, a group of seven nala and two female medical stu- dents and laboratory worken with aRa ranging between 22 and 43 years and with different smoking habits (2-3 cigarettes to 2 packs daily) were aded not to smoke during the sia houn preceding the experiment. Cannulae with 3-way stop cocks were utilized to draw blood from the mcdian cuSilal vein a1 0, 7, and 15 minules afler a cigarette was smoked for two minutes. Nico- tine ksvels of 60 to 67 rp/1 were found iw subjects 15 miww/es after smoking one standard cigarctte. 71re technique reported hcre, which is very rapid, reproducible and sensitive, facilitates the determination of nicotine in smoking subjects during the actual proces of srnoking. Cotro. A. et d. Cl/nk. CIYGnk. Act.9S(7):47)-4t1, 1979. Other aMPprt: American Tobacco Company. From the Hormone Research L.boratory, Department of Patholoty. Uni- versity of Miami School of Medicine, Miami. Fla.; Division of Bk.chemical Pharmacology, Medical ('oik6e of Vir4rnia. Richmond; and Union Carbide Research, Tarrytown. N.Y. A TOTAI.I.Y AUTOMATED SYSTEM FOR EN2:YME IMMUNOASSAY OF THEOPNYLLINE IN SERUM TAeophynine is a naturally oeeurring alkaloid which has long been used to treat bronchial asthma. Since the margin between its therapeutic and toxic concentration is very narrow, many techniques for aeeurately monitoring serum thcophyMine have been described. The one reported here, however, has serveral hitherto lacking advantages, namely high speeiflcity, simplicity and automated speed. This particular assay is baacd on competitive protein binding, with glucose-6-phusphate as the labeling agent and a theophylline antibody as the specilic binding protein. All phases of the assay are totally automated in a Kinetic Analyzer (KA-130). Shifts in the absorbance of NADII at 340 nm reflect enzyme activity and art directly rrlaled to the drug concen- tration in the sample. With this sys/em, as many as 75 10 pl samples per hour can be .ssayed, and results are availabk at )0  intervals a/ter initial sample preparation and preincubation. Upon comparison of measurements obtained on 1)t clinical samples with a manual ultraviolet speetropholomelric method (s) and the presenl method (y), the slope of The comparison curve was 0902. The re intercept 0.402, and the correlation coefficient 0984. The variation coefficient /or duplicate samples run on the same day was 4 9%; for samples run on dillcrcnt days, it was 0.1%. In summary, its prcvionsly noted specdkHy. sensitivity. spced, and simplicity, plus the small amrninl nf reagent I I t I t it rer,uires, 811 make this technique an attractive alternative to chromatographic procedures, especially in emergency aituatiora and under heavy work load conditions. Cartro. A. et af. Cllnkd ClieerLtry 24(6):944-946, 1978. From the Department of Patholo6y. University of Miami School of Mediclna, Miami, Fla. ANALYT1CA1. A1.TERNATIVES TO RADIOIMMUNOASSAY: A REVIEW In recent years. radioMnmunoassay has become one of the most rapidly growing analytical techniques used In the clinical laboratory for the detediom of minute quantities of steroids, hornwnea, peplidea. drugk and other wrb- stances in body fluids. This procedure Is highly sensitive and apeciAc; however, it has several dis.dvantapea„ such as radioactive ha:ard, waste dispad, IioenainR. FDA regulations over instruments and reagent production. and tM short shelf life of reagents. Analytical alternatives lo radioirnmwronar--ta6 ddection mNhod, light scattering and slide-based lechnir,un, and isodemity anairsea-have thus been studied to determine the feasibility of developing new approaches for the measurement of mirwte quantities of aubtruroa tha/ could be free from the above inentioaed restrictions. In all the alternative methods reviewed here. the use of radioactive material is avoided. Easynw irnmunoass.y. especially homogeneous immwsoassay, Is so far the moN prom- isinR one studied. This system offers a possible saving In time and mrorey, and an increase in safety with no deterioration in sensitivity or accuracy compared to radioimnwno.wr. The relative simplicity of the end point deteelor, long shelf life, and the tcorarnica of this assay make it attractive so mo.t clinical laboratories. Cart% A. et r. TbtCllnk.l Report 12(9):7-18, 1978. From the Department of Patholopr. University of Miami School of Medicine. Miami, Fla. COMPARATIVE DETERMINATION OF PItENYTOIN BY SPECTROPIIOTOMIrI'RY, (TAS CIIROMAT(XlRAP11Y, I.IQUID ('/IROMATY)GRAPIIY, ENZYME IMMUNOASSAY, AND RADIOIMMl1NOASSAY • In this methodological paper, sera from hospitalized epileptic patients being treated with phenytoin were analyzed for the drug by speclropholometry, gas chromalography, radioimmnnoassay. enzyme immunoassay. and liquid cMo- ma/ography. The fundamental differences in the five methods lie in the method of extraction of the phenytoin. When the assay values obtained were inter- computed statistically, it was seen Ihal, of the five methods compared, there 77 76

ia suRkient correlation among four to make their differences clinically insig- oilkanl. Only radioienmunoassay appean to have a poor correlation, giving relatively high concentration values consistently. Judging from thrx teata, enzyme immunoassay and liquid chromatography appear to be attractive alternatives to the more traditional methods of spectrophotometry and gas chromatography. Indeed, total extraction analysis times of 20 minutes, coupled with simultaneous determinations, make liquid chromatography superinr to all tbe traditional gas-tbrornatotraphic methods. Cartro. A. et r. ClinkdCA.mlrrrry 24(4):710-71), 197l. Proro the Departmeat of Pathotogy. Uaireniy of Miami School of Medicine, Miami, Fla. Vtt Eptd.wsto[ogy PULMONARY FUNCTION IN H6TEROZYOOTFS FOR ALPHA.- ANTITRYPSIN DEFICIENCY: A CASE-CONTROL STUDY The purpose of this prospective study is to determine whether the heterozygous condition for .,-anliuypsin (AAT) deficiency is associated with Impaired pulmonary (unction, accekraled lung aging, and Inereased risk of emphysema. In the present paper, the initial crou sectional data of a longi- tudinal study of AAT heterozygotes and eoatrol wbjecls matched for age, aea, smoking history, and ethnic group are presented. Using a case-control de.i=n. the eases included 37 heterozygotea for AAT de0ciency, proteae inhibitor phenotypes MZ and MS, selected because they were parenls of children with homozysass AAT deficiency identified in a statewide newborn screening proitram between 1971 and 1974. Ap o/ the helerozygotes were len tbaa 40 years of age. The control subjects were selected from a random sampk of a working population participating is a longitudinal study of hmg aglng, wing a 2:1 match of enatrol aubjeeta to AAT deficiency cases. Test procedures used for this aludy were (1) a respiratory symptom questionnaire. (2) apitomdry, and 0) the single-breatb N, lest. Tests of pulmonary wrochaaks wera also performed on the easa, but not on the controls. Results showed no dilference between the cases and control aubjecta, either in terms of respiratory aymptoms or in ternr of puhnonary function data. It aeems, there/ore, that the heterozyllous phenotype (Pi MZ and MS) is not a riak factor lor impairmeat of pulmoaary function up to 40 ye.rs of age. 1r4t, A. S. et d. Anwk.n Revkw o/ Rerpberory Direare 120(4) •739-766, 1979. Other urPprtt National Fleart, Lung and Slood Institute. Prom the fkpartmenu of Medieina and Physiobp. Universily of Oregon Fleafth Sciences ('entrr. Poruland I A STUDY OF SMOKING AND PREGNANCY W1TH SPECIAL REFERENCE TO FETAL GROWTH The Influence of smoking o0 pregnancy aad Its outcome wr esamhed in a prospective study of 3,272 urb.w woasen, or 90% of the 1974-75 pregnant population. TM fetal bipariet.l diameter (BPD) wan measured u/trsonicatly beginAing at the 19th-201h wroek, though early ia tba study these measurements were first obtained at 18 weeks. Separate SPD growlh curves were coestructed weekly for the womea who lraoked (49%) aed for those who did not (S1%). In 45% of the casa, more 1h.m one flPd aseawre- ment was utilized to construct the growth eutrea- Only those wqnea who delivered more than 266 and prior to 294 days aiece their tast nmeeatrwl period were lachrdcd so a/o avoid erroneously akewed powth prolrw. DurinR gestation, the SPD increased nars rapidly among the aoo.awkere thm among the arraktrs, the difference being aigniReaMK apparent froaA the 291b week onward, and correlated positively with the average awnber of cipr.llea smoked. The norrnsokers showsd a greater leadency for pt.celampsia, wMia the women who smoked had a higher proportioV of abruptio placentae, prt mahxe deNvery, and babies who were aas.ll r« their Restational age or bad a low birth weight, short birth Mwph or eMwdl baad ckean/erence. TM low birth weigMs s.ere iadependent of the wro/hen' weight before pregnancy and of their subsequent weid+t piw. In compariwa with data obtainad /roat a similar 1963-64 aludy of the eame conwrwniq. the proportion of pregnant womee who smoked inereased from 44% to 49%. The present study dcmoa- s(rates the early onset of iMrastleriae RrowtA retardation ia prepn.M molbets who amoke. These results suggest that thit aNnw from a direct pharmacologic effect on the fetus rather than frmn nutritional deprivation. Also inAieated is ~appareat Inefficacy of the program aimed at deterring women lrom L Perasoa, P-H., Oteennert, L., Oenmer, O. and KrRader, S. Act.OAstetricl. et Oyneedogin Scanl(wwk. Suppl. 70:22-79, 1978. From the Diagnostic Ultrasound Laboratory at the Departmeat of Obaietrfcs and Gynecology. University Hospitd, Malmi/, Sweden. . CHARACTERISTICS PREDICTIVE OF CORONARY HEART DISEASE IN EXSMOKERS 1lEFORE TIIEY STOPPED SMOKING: COMPARISON WITH PERSISTENT SMOKERS AND NONSMOKERS Based on data compiled frorn three or more Kaiser-Permanente checkups of white and black men and women, easmolten and continuing unoken were compared for a large number of char.cleristics associated with the risk of corrmary heart disease (CIID) in a population. These characteristica were assessed when .1/ were smoking cigaretles and those who remained r.on- smoken provided an additional comparison group. 7 he ones who later auit demonstrated statistically signi/icant di0erences from the persistent smokers in certain cardiovascular sympoms, social and persond eharackristia, intensity of snmrking, and some other trails. In addition, in most or all race-ae. .ub- groups, they had a higher relative wei`ht, consumed kss akohd, smoked fewer cigarettes for a shorter time, inhaled kss, had a higher vital capacity. 78 79

~ lV ~ r lower .ktukocyle count, lower prevakeooe of abnormal electtocardio2r.rns, ku exettional chest pain, dyspnea and leg pd.ti, higher educational Ie.el, aod a tendency to answer a psychological questionnaire less like penoas w(so later developed myocardial iefarcts. Althoullb they were statisticdly sip.ifkant, most of thae diflerences were nevertheless quantitatively small. Cholesterol levels and blood pressure showed only small diRerencea. Adjusting tSe data for smoking quantity had little RRect, for eertaie characteristics that seemed unoking-related, on differences betweee t!w per.isteot quitten and the per- slatent sewkers. Por several paramelen, " bdez ezaminatioe thQse who quit were more similar to those who aever eteoked than to pefsiatent nrwken. Compafison of the outcome of CHD amo.R esrookcrs with that among persisteM smokers should account for smoking Intensity and other baseline cbaracteristin that may affect the degree to which giving up smokinn might ahtr the risk of CHD. F.Iedrn.n, G. D. et al. lorrrnd of CA.orJc Diseases 32:173-19Q, 1979. Other aurprtr Kaiser Foundation Research Institute. From the Department of Medical Methods Researcb, Kaier-Perrnanente Medical Care Program. Oakland. Cal. THE FINNISH TWIN REOISTRY: SASELINE CHARACTERISTiCS. SECTION 1. MATF.RIAIS. METNODS, REPRESENTATIVENESS AND RESULTS FOR VARIABLES SPECIAL'f'O TWIN STUDIES This publication describes and documents the baseline characteristics of a questionnaire study of a twin population derived from the Finnish Central Population Registry. The main portion (sections le and 111) of this report outlines the rexrhs and distributiom by age, aest and tysosity of most of the variables inchrdcd In the study. Oe the whok, only those twin pairs with both members responding to the questionnaire (about 75% of all twin pairs in the rejiury), and generally only those whose zygosity could be determined (93% of those meNioned above) were inchsded. Comidercd in this section were the particular variables that are spedsl to twin studies: eoatac" between twau, birth order and handedness. The fdlowieg eonetusione were drawn with respect to the populatioa^s represenlativeness: (1) because of tnortatity, the otder age-groups are undcrreprcsented, since the probability that bolh members of a twin pair are alive is lower than the corresponding probability for one person; (2) the population is not representative with respect to marital status, since there are more single persons among twins than among dnglctons; and (3) there arc more rural inhabitants among twins than among aingktons, either as a result of aekective population migration to urban areas or because of a higher twin birth rate in rural areas. Kaprio, 1., Koskeavuo. M, Artimo, M., Sarna, S., and Ronrasdo. !. K.nranrirrrtrrarrrrn tutIM1Y+0 M 47 I 74. 1979. f'r,.m ihe tsrp.rrnwnr of PohIK Ilratth 5cunce. Unrvrnd7 of Hclsinki, 11rl.inte ru l I I A DEFINITION FOR THE COMMON-FACTOR ANAI.YSIS MODEL AND THE ELIMINATION OF PROBLEMS OF FAC7OR SCORE INDETERMINACY Mostly with mathemalip, a rigorous definition for a factor analysis ewdd and a complete solution of the factor store indeterminacy problems are pre- aented here. Such a model consists of a random sequence of variates deened on a probability space and satisfying the usual descriptive equations of the common-laclor analysis in which the cormnoa-factor scores are dimensionaqy Independent. The condirions necessary and sufficient for a model to lzist with essentially unique, and hence determinate, common facWr scores are givee. Also given are parallel resulb for the esistence of nodeb with eomsnique, and hence indelerminNe, scores. That two models cannot eaist with esxntiaqy unique hut different scores for the same factors in then proved. The meaning and appiicatioe of theae reaults ate discussed. Williams, l. S. (Crrnrp.c4rr, D. W.) lsycliornetrl4a 13(3):293-306, 1970. From the Department of Statistics, Colorado State University. Port Collins. REPLICATION OF THE FACTOR STRUCTURE OF THE COMREY PERSONALIIY SCALES This report summarizes some of the more interestind results of a questionnaire administered in a pilot project run in preparation for a study of environmental and genetic escton in smoking behavior to be conducted Ie Sweden. The factor structure of the Comrey Penondily Scales .ras replicated in a sample of suburban Denver residents (N=377). Correlations were oA- aerved between the Comfey seakts and age, socioeconomic status, ao, body lype, a short locus of control scale, the Raven Progressive Matrices Test, and the Neuroticiam and Eatravenioe scales from the Eysenck Personality Ques- tionnaire. The correlations observed between aelf-reported leisure time activities and nervous habits provide a partial empirical validation of the Comrey Activity. Slability, and Eatravtenion aeakea. Vandenberg. S. O. and Price, R. A. (CrumpsrAer, O. W.) PiyelwfoRicd Reporrs 12:343-352, 197d. From the Institute for Behavioral Genetics. University of Cobrado. Boulder.

Active Projects FoUowiog is a list of the principal inresti6aton, or mstftatioo., d projects under way or activated in the period since the prerious lZeport, toaether with the respective project titks. Completed projecta ue listed in a later sectioo. PRINCQAI. INV Ra17GATOR OR INS'RiUT10N MARIO D. ACETO. PM.D. AssocLte .re.Nt. Udtf. RkRso~~~~ JOSEPH C. ARCOS. DSc, hofiuor o/ Medicine. Trlane Uni.ersMy 5ct+ooi of Medici.e. Ne. Orleerr. ALAN K. ARMITAOI; Pu.D, ltese.rcA Directo.. 11.alelost Lalota/aeite Ew ~. ILrrople. Nanl+ Yottutrire, Eft - MARII.YN 3. ARNOTT (RASCO). Pw.D.. Arsbnwr Aioioou aw1 Aw.rw P.o/ersor of siol.op. The Universily of Tea.e Syaem Ceneee Cenle` M. D. As- denoe Ha.pitd ud Trww InwNwe, Houpon. LESLIE DABR. M.D, AswrWe hoJe} Cdkp ott PMlrsici.ae i Surseo.e. Nterr Yoel. WILLIAM F. RENF.DIGT, M.D.. Au4- unr ho/tssor of rdiNrk+L Ur,ivenilr of SoMtber. Cali(arwi. Sebooi of Medi. aint. Di.i.iow of IlswNolo{r and Med. kd OeaeticS CI+il6reaY HoyUd ot Lae Myely Los Anaeks. SEA J. .an den oHRO. M.D. Research Pedionklew Ad/rwn ho/rssor lw !b. sraluks. b.isasilr of 4Qaldania Sct,ool of Publie Hee/ik Oatl.W. RICHARD /. •INO. M.D.. rro/essor of Mdklwe. Uaiverwr of Sowt+ern ('ali• /oreia Sct+ool of Medicine. Los An• teks; Ylsbinff Auoci.rt in Sbwrrdic.f Fws(weniwt. Ce/iloasie Insrirwt of Teet•noloar: Direcror of Cordiolufy owd fwir.w.wol Mtdu Ant. Hunlinsion Mcmaief Hosplal. Psedena. Cal. PRO7IiCT 'IT11Z 3uroawpecMe binding of elcolla. AcU.alina maabolMer of dletflyl nl cyclic .Mrvriwlw a.A poe11w of /M reqome 1e eesp.w Mduoen. Re1Nlo.~ eRiP lo.-.Mroe..f.s eta Rron) abeotpion o( l.belkd .ioe/ir by IM anesuerised eti1 trerw eipr a.wis as wep.red to ttul tras sipreew HyMoc.rbon redabdiei.a earyma .d 1wy caneer a awotMy I. aa~We2 se .d n«un.i.. erbkc+e: blood reaK ddoelero.m e.l •i M a.rtarwMloa, oIMy PruY P~w~dh My of dpreae...t. Mdyn.d uartor.ralloR ar,wyewnM and po.eaee .aiviry of elauene re• lued .waerid and lhe k"Poee.eea of e Platiea traeliow (Pf:U I. d+ese Pn.w- elere Prlaronat) tuncliom uMe In aAoleecuMa and Ureir yeenl.-aw epiAewdol"k aMroac\ IaRlbNios of elol.rerd aMeke by 7. 11/1oct+olrlerd EOed ot carbon na.oaide an droleMtrd wM.Aolisru ot rbe ener W.dl Clwksterol inbib8ion awd eerAow monoa- ide end atAerosckrosis 83

PRINCIPAL tNVESi1GATOR OR INSTITUTION FRANCOIS M. BOOYSE. PN.Q. rru/rs- sor of sior Armisny. Rwh-Presbyterian- St. Luke's Mcdicd Center. Chicago. (Now Senior Inresrir.tor. Michael Reese Research Foundation. Chicago.) RAYMOND BOSSk Psr.D.. Auociue Director. Normative Aging Study. Vet- eraes AdminiurNion OWPatient Clinic. Boston. DAVID L. BUSBEE. Prr.D.. Associate Professor of liwdorr. Drpwtmrnr of tilojorird Sriencer. North Teaas State UniversMy. Denton. ELROY T. CANTRP.t h Pit D.. CAdn m.n. Drprtwrenr of M.rmwcotorr. Teaas College of Osteoynhie Medicine. Nortk Teaas S1are Unrversity. Deaw.. ALBERT CASTRO. PN D.. Director. Hor- nrowe Rese.rcA LAu.erory: rro/easw of Parlbfurr en/ MrJsrrne. University of Murni School of Medicine. Mumi. 1-la. PROIECT T1TLE rRINCIIPAL INYESIWATOR OR QN41Rf11TlON FROIECT TIiLZ in riro and /n vitro responses of endo- tse)ial alls and tAe vessel wall to nico- CARLTON K. ERICKSON. M.D, t ro- Blood-br.in tnoaitoriag of sustai.ed .ko• tine /nsor of rArmxolorl. The U.ivenity ti..lerela is r.ta of Teaes College of Pr.ro.cy, Austin. V. GENE ERWIN. Prt D., t ro/rssor of Actions of .iooti.e om isolated perf.sed A amoking research project in the Nor- .+~y: Dean. University of wouse brain wNi.e Aging Study ~~ ~ hod of Ph.rwucy. Boulder. The biochemical and physiological char- seterisrin of a potein rhich .peci/l- capy binds polycyclic Moma/K hydro- carbona Aryl bydroearboa hydroaylase im single alls and wbPopd.uiau of human lym- Phocyle; sutd other ceUs Nicotiue in Wood: detection by radio- imnwno.ssay I WALTER B. ESSMAN. M.D., PuD, Professor of rsycAnlop .nd eloAlem- brry. Queens College of 1M City Uai- •erwy of New York. Flushing. WILLIAM H. FISHMAN. Prr.D rra/- /rwr.l../dta Canar Ra.arcA Po..da- tioS t.. loUs, Cal. JUDITH ANN FOSTER. Pw.D, Asroe4 ere rro esor o/ AioeArndury. Udher- sity of Atlwe. JACK W. FRANKEL. Pw.D. Associate rro%asw o, Medical MkroAArlory University of South Florida College e~ Medicine, Tawtpa. FRANCIS C. CHAO. M.D.. PN.D. Sen- Matek/ activatiow and blood Aypercoasu- AARON E. FREEMAN, ht.D. Scientld. for fnreulrero., Center for Blood Re- /abilMy La lolla Ca.cer Reaearc! For.d.tio.. searclh Bo«oa La loRar Cal. CHARLES 0 COCHR ANF M D. Mem- The medialion of in/amrnalory injury of LARS FRIBERO. M.D. Professor .nd Ier. DeP.rrmrnt of Immrnop.rAolor7. tissue CAsirmeR, Dqartmasr of Ewrbow- Sn Clinic and Research Founda- wrenai H ~lewe. T.e K.roli.ska Iswi- ~ tion. I..lopa, Cd. ltrle. Sloc haM. ALLAN C. C(H.LINS. Prr.D., Associ.re Oenetk analysis of .eurochemkal and be- KIELI. FUXE, M.D.. Associ.re Professor Professor of rArnserdosy, Institute for havioral effects of nicotine and alcohol of HLtofcry. KaroUsrk. Iwstilwe, Be4viord Oe.eticti Usdversily of Stockholm. Cdorado. Bodder. MORTON OALDS7ON. M.D. Associ.te Professor of Mrdicinr New York Uni- PAUL T. COSTA. Jr. l1e.D, Associate T1e relaliorr between smoking motives, , New York. veraNy Medical Center Professor of lsycAoloty. Univeraity of persondity and feelings . Massaciraette at Bostost Dorchester . . MICHAEL C. GEOKAS. M.D.. Pu.D, Professor en/ Yice-CYubmas. De~.rr- IRVING P. CRAWFORD. MD.. rro%r- l.eekoeyla dastase<ontekmenv interae- wreM ot Medcine. University o/ Cali• ror .wd Ck.irnrn. Dywtmrnt of Mi- tiotq iw (Se etiology of empb)sema /oreia School of Medicine. Davis. croi/dory. University of lo.a College of Medicine. Iowa City. TERESA OESSNER. Pst D.. Assnciae Hedth Re- C.nrer RrurcA Salenslrt DAVID W. CRUMPACKER. PN D.. ho- Genetic and environmenid factor af- . RoeweU 1'ark Division, B+t- search Ine /nsoi owd CAoirm.n. DeO.rrwrrnt of fecting smoking beYavior » Ido Ewrirowmened, ropdabn and Orro- . w4mk lioforr Untversity of Cdorado , . Boulder rACQ1)ES E. GIELEN. PuD. Assiswnt . rro/essor. LuGor.rory of Medical ROBFRT Fl'IIT PN D Professor of An.- The eRccts of hypoaic hyppnia carbon Chemistry. Todrofory end Nyrirne. . . tomr. Mahipn State l/aiversny Fast . monoside and Irealmrnls influencinr InslNule of PatMrloky. University of. . Lansing. . bypoaia tosicity on enducnm like cclls l.ftle. Liere. Belgium. in ainrays of young rabbits Correlatea o( streartob.ceo rde lala• .aiu. Markee ProQe is bowcY.l s.uoo., and oao}Irr of a.rokers and .o+s..okars Isrrdvra>ye.1 of daatk Bbwe 1. hy dlea.n Suat~ su hsnR eaneer: diapoqk le.l r Persons at high rhk Rode.l..d Mrwaa lunf eptheli.l aU w4 twt r.lool /or earanokewe.l. rese.rc! M elln. ~ M.dka oa tlr SweAlr~ Nkoliae eatecholamines ..d tnsrroe.doa rrine fradiom Biocherwkd b.ais of pediepskion to edronk obetnrctive psrlmo.ery diaew Circrduirt{ pncreatk elasu.e 2 and a.- physerna in ww MrumaeoRenetlee of eo./uplbr and /rp cancer risk Cigarette smoke and polycydk •y" earbon anelabdism ie l.M end kidnq Modulation of u~1 hydrocarboh hydroay- lase and ePoaide hydratase in anuwal tissues and in cell cultwe by cigarette srrwke condensate and other chemkala m as

rRINCH'AL INVFSi1GATOR OR INSTITUiION RONALD W. OILLETTI; MD., Direc- ror. Ieuk Srknce ReseorrA Unit, Caw ar Center of Hwaii. University o( H...ii et Me.os4 Hosolul.. PROJECT TiIL!< FJled of tobacco byproducts and aber etnkawnenrtl tonlemaunrs oa lyee- Pboid cstl bmi.R .ed fu.cuio. 1'RINCQAL lNVfallfaATOR OR INSi17lJitON CAROL 1. HENRY. hr.D., Dirrctor. De. hwent of Eqerlnrewaf pnrofo~y ~o~ctl AatocWe.. l.e. 6.- lRO1R+t.T iTRi 8vdrrioa rd ei.r.elwia.e{o, d r.4 b.Rnw elutio~.w.P aym ae.wre of Ni Immossim, p~ Di.NoA~ ed.s.ys l.diroed by eAr- ~~ c\es.kalas.ol. OERALD 1. OLE>CH, M.D., ConnJunr i Hu.us ftyDersensiUvily so treli=cee from HERRERT 11. HERSCOWITZ. tr.D EQec1s of eig.relle awsote ta wrs 1b M Medk ee, Research Laboratory for Iob.coo ., Associate Professor of 1/ p a. . in.mrwnre .wer ewc+io W of w Allergic DiseucS Mero Clfnk artr OeorRelo.ha Univenily Jcbols- . T P - R" .w.wy elveolw .r.r:o a Fou.dslion; A,.xi.te lrofea.oro/ lo- ki.e and DeMlstry W.shirpo.l D C o ep tnnd Medkiwe ewd /nnwrwelory. . . . Meyo Medical School. Rocb.aeer, Miar.. JEROME L. IIOINACKI. PM.D, Awferw Cigerelte sn.oki.g ..d high de.eNy yp. lrofr,.or of Iiofogkal Scknc»e, Ua+4 KOleiVA JOHN W. GORROD. DC.C.. Lernrer Iw Toaioolnk.l Mudks ot• 'pyr{diee-Noa- .ersMy of Lowell. LoweR, Mess. A/oPA.rNUcy Cbelsee College Univee- ides' wd some related cornpounM . . .i1y of LondoR London. WAYNE H()5S, PM.D. Ard,aM ho- Jerdiae of niooline wertdbr with blood , Irr.w. Cewler /or bre~n Resewcb, 'i1e aM. UniversMy of Rochester Medical Ctt HODA A. OUIROIS. l1M.D., Associate AHH ud rel.led irnnw.ocbemkd mwt- ler. Rocbe.lee N.Y rrofr,.w of C.anrnrwiry and Ew.irorm- en: a tusoepibilUy iadea for lunR , . wrewbf Aledicinr. Universar o( CeH- wcet ALPHONSE /. INOENITO, M p. A.r- Aelie.w ot cerbon wosoalde .mid 1eMoe. tornie Cdkp of Medkine. Ir.ine. clre trofenar of rAwn..rofverE.ri on Mia+.l welebolir ../ t.w Cwali.. U.'wwsily School of %Ney. ~ HIRA L GURTOO V Sc D M MV Role of genetics and oolywarnelk hydro- dne, Oree.viMe, N. C. . . . .. . .. . Pn.D. A,sociur Cewrer Re,r.rcA urbw metabolism in Mrnaa lung eu- I Screnritt, Drp.rtmrnr of E,prri.eesirel cer HARRY L. IOACHIM M.D. AnendIn~ The bed irenwnt respo..e is lry ee.at TAeroPrutkr Roswcll Park Memorial Ir Le.w[ HiR HoqMC ~ , Institute. Su1eb. U h euor rr dd.ati A CAROI.MP. B. HAIl., M D.. Associate rrofnuw of rrdierrlr, ond Mrdrrnwe. Universiry of Rocbester School of Med• kine, Rochester. N. Y. LINDA M. HAIL. Per.D.. Assistant faro- fruor of fiioGrRy. Mess.chuselu Inni- twe of Tecbnokyy. Cembridge. (Now Anociore Irofeaur of (:ewetk, ond Nrwosrknrr. Alberl Einstein Cdkfle o1 Medicine of Yeshiva Universily, The grona, N.Y.) MARGIT HAMOSH. PN.D. Rr,e..cA Associate. Drprrrrwent of 1 AyaloloRy and siopAyskr. (korpelown Univer- sity Sebools of Medkrne snd Denris- 1ry, W.sbindton. D.C. PAUL HAMOSH. M D., Auoci.te f'ro- Jnwr of rAyrlo/oty and RwrvAy,k,, ond Medicine. Georgetown University Schools of Medicine ud Demislry. Weshingtotr. D.C. NORMAN W. HF.IMSTRA. PN D.. rro- frnor of l,ycholotr. !)orrcror, Ilrn+en !:'et rort liAo.ero.y. tlnirerfily o(:Sourb hslote. Vermilhon. TAe relationship of lower respiratory tract drseese in infancy to rbe development of chronic lung disease in adults: devel- opmem and rime course of physiologic .bnormeliuea indiwlive of early ob- uruclive airways diseesa Genetic diQeres,ces in wieoline sensitivity i. DroioPAib nrelewogautr strains FCriaM.l lung devdoomat. metabolism a.d eiilweue woke pleel of smotinR on evolwi(. of the rwainnrr eaprdory low-volume curve Sorne behavioral espects of .muling and uswting deprivation S.rReorr, Ne. Yat. AARON IANOPP t*..D., 1•roftaser of trAofop. He.~lb Sek.oes Cemer, Suls UnivenAy of Ne. Y«t at S1oq erook, Sa.y Rraot. DAVID A. IOHNSON hr.D.. A.+/wnt rrofetaor o elochewrj,try. East Tes~ see S/ele LnivenNy Collede of Med- ki.e, /obwson City. WILLIAM 1. JUSKO. hD., Associate 1 rofessor of rArn.ecertks; Director. Clinkd t Arn..roal/nHks L.br.toer. Millard Fil4wor. Hoqild, Mlela EDWARD L. KLAIdER, M.D.. Senior SclenNst, The Worcestsr FoundNion for Esperinsewel Biology. Ine. Sbr.ws- bsuy, Mass. JEROME Kl.E1NERMAN. M.D., ho- frslrw and CA.kieew, DeprtwreAt of retAoGrar. Mount Sinai School of Med- kine, New York. KLAUS E. KUETTNER. Pe D., farofeuor of fiiorhewsi,try In OrtAoprdk; Deprt- erewt of SiorM.niury, Rush College of Hcalrb Sciences ud Rush Medical Cd- kge, Rusb-Presbyrerien Se. l.uhe i Med- kel Cenler, Chicago. 11 q 1 rer eof Proterw {.WibMbn by dr ;w.r of hry elstii d.- e!do{e ~~ k enrcy.es sid M Effw dYil u: ••• w ossWlo. .. Behavior end blood'res.we M r.b ta4 b..inil pwfn.W .nd pMWd nioollrr ta/OeYrt DiMribuUon wd .eqorw of wnMe ~r+twsor upakt end daewboay/dbn W bod- (APUD) cells nwd aoeuroe~ ke (NPs) M wenNwel/u I oe.l reAul.lion ot worwal rrwd PNbolobie invnioa e7 86

PRINCIPAL INVfST1GATOR OR INSi/M1ON ABEL LAfTHA, Pw.D., DireHa, New Yorl Stals Research Institute for Neo- roclaemilrry and Drug Addictim New York. li CLINTON LAWRFNCP. M.D., A.rU- t.wt Prn/e.ror of Mr/kiwr, Raylor Col- kge of Medicine, Hou.loa. JAMES C LEH hr.D., Aul.t.nr Profta- tur of F/orArndrtry, Saint Louis School of Medicine, S1. Louis. JAY A. LEVY, M D, Assistant Cllwkd Pro%saor ./ Mriiciwr Research Auo- cr/ae. Cawaa Rerearrch InMi/ule, Ud- versity of California School of Med4 eine, Sas Fra.cieco. PAUL D. LEWIS. M D., Srnlor Lrcrrer in Nirror.c/1ul..Ry, Royal Postgraduate Medreal School. Nnw/ncrrnit[1 Iloep- ta1, l.oafow. IRVIN H. LIF.NF.R, PIrD, Professor of SiorArmtstry. Uni.eraty of Minnesota. St. Paul. HENRY T. LYNCI/, M D, Professor ewd CA.frn..x, Drpertn.rnr of Prrrrs. Nr. Mtdklwr and PuF/lr IPr.nhA, Crei&lMo. Uwiver.it>. School of Mcdi- cine, Oreah.. R. RUSSFLL MARTIN, M D., Professor of Mediciwe, and Mk.olioloty t Iw.- manotosf. Saykw College of Medkine, Hawaa. REGINALD O. MASON. M.D., PN-D. frofruur.wd CA.Ynww. Drprtrwrwt of rrArlfop University of South Florid. CoRe~e of{ Medici.e, Tamp. (Now at 7 Ae U.i.eniy of Utah Co1kRe of Med- PROJ[Ci TRL6 ERect of materaal .moling on brain de- vekpmenl in the oll.prins ERecU of cigarette smoking on immuno- /a production by human broacbial rinNacytes PJeets of lipnds on Olatekl mkrotubuM wewbly Ponlble genetic determinants of chemical tarY{.Opc/K.l. E/Iac1e of tobacco dlabida on cell Oro- duetion in the developing brain Proleaee iabibilora attac6ed to micro- spheres a. a potential therapeutic aae1M for emphysema Stetirlical-lenetk evaluation of risk /w ton in lung cancer Induction of uyl hydrocarbon lydosyla.e in lymo6ocytes and pulmonary mKro Phat« EAee1s oI smo\-u1~ awd nkolinc on growl~ a.d fdio. ot hwaaa endotbclial oelb a PRINCIPAL INVldflGATOR OR INhT1TUi7pN 1. WISTER MEIGS. M.D., Clinkal Pro- lraror of Er'JrndofoRr; Dirrnr, Cow- wrrtKMt Cancer E~i/rmlolury Uwit, Yak Univereity Scbd of Medicine, New Haven, Cora- MICROtiIOLOOICAL ASSOCIATES, INC» Bethesda. Md. J. ANDREW MITCHELI, Pw.D, Aac4- tent Projetavr of Anerowrr, Wayne SWe University School of MNici.e, Detroit. FP-RID MURAD, M.D., PR.D. DMertor, Division of CUnkrl PAre.ecoloq, Deparuwe.t of Internal Medicin., U.4 venily of Virginia Sclod of M Aici.e, Ch.rloltavipe. DONALD 1. NELSON, ht.D., A>ai.r.nt Professor of Chemistry. Clark Uwiver- .ity, WorceYer, Mess. FRANZ OP.SCH, PN.D J'r%nor of PA.rnl.rology; N.~.I, frct/ow on Ilo- cAe/wirr r6.rn..eofary, UnhetsMr of Maiss. Main; Wal Oasaar, SEVf•RLY PAIOEN, h1.D., C.ww Re- tt.rrA Scirwtbt Y, RosweR Park Me- morid Ilcstiluk, suffalo. PROJt,CT TI'I1.rE Review of lung cancer in Conneetkul, 1f73-Oresent 3mote lnhaWion cardeo~e..eis st.dia M mioe Standardiratiow of .r.ys 1o tweawlre ary1 hydrocarbon Aydroaylsae i. M1.am tiplrea CoRabo.alive sludy with Dr. Jay IsvP (University of California San Pra.cMoe Medical CeMer ) ded`ncd lo delnolee 1he Rewetie control of aewouoyk virw eaoreswrw in hybrid slrais of mka Imilaaluiom a+d Werl.e fr.oion ia tb rr Mech.nism of nNrk oaide adhatio, af pawylate cycl.re Speetroaook audies of 1he iMeraetbs of choliaergie lip.de with nkachfe ra- eqlor proteiws aroe of epotide hydrMas., Yy" dehydrolten..e and em.)uww e*- symes i. Ihe conlrol of .wtye.fe aad Iran.formirae roetabdiles ol polycye/k Aydroearbona oee.rrirlll in eig.rdle swlote a.d inrerladivWual oo.yarieon of their activities in maw Geeetk wsceplibility to bl.dder ca.oer kine, Sall Late City.) KENNETH PAIGEN, M D., Dirrctor, A 1ed of Rlucarowidaw /M b!d- OBRAI-D R. McC1.EARN, h.D, Dl- Heredity and tobaccorelated behavior In Drprtnwnt of Moderu/w fiobofr, ocr ea ncer recror, /nrNtwe for Rrherior.l Ge- the mouse Ileedth Re.earcR Inc„ Ro.well Put wetkr, 1)ep.rtwlrwt of PrycAolocy. Uni- d l d f Pl f C Divisiow, su11a1o. ver ry o ura ar- o o School o maey, Boulder. DENNIS R. PI?TFRSEN, PM.D., Aul.rant Inlluene n of Reewype, sea and cilrowk Professor of PAr.e«ot. . I y e are /le snldinR w nkdine and al- HERBERT McKF.NNIS, /r., PN.D., Pro- leasor of Phrnwe.kKy, Medied Col- Cooperative studies allned at the develoo- ment of immunoasraye for nkotine and of ('olorado School of flqrmacy, Booklet. c metabdi.m io mice 0 k" of Virginia. Vk~inia Comnwe- wealtY Univeniry, Ric6n+ond. nicotine metabditea CARL W. PH!RCI; M.D.. ht D.. Profta- Biology of wppreewr T oeR. tn rr of Pathology .wI MkroMr/lorr•hn- 0- n/unolory W.ahin ton University m HAN3 M/ lf R. D V M.Trnl.v Srs Sri. ri r Ib 1 1 1 h S Trawsplacenrel e4ecle of nirrosocom- d , g School of Medicine; Perholoslst-lw- tD J N tn .rk, rn , c /..n c ec a NNwy, at II./IVN, Me m . poun CAkI. The Jewish /lorqtal of SI. Louis. St. LOIIIf. 89

PRINCIPAL INV[H•11CATOR OR WSiTillTlON ILARI RANTASALO, MD, Professor nld Chdnw.u, Drlkwlw/ew1 of b.lt4 HetllA Science. Unwer•i1y of Helai•tL HdeiMi. Fidand. RONALD E. RASMUSSEN. M.D„ A} rociae A!/rwct Ao/rswr b Coawiw.• w/y.wdEwrioxn.rwtdbIelklwe Ui~ r ~ or.i• Collep ef HERBE,RT Y. REYNOLDS. M.D. M- /raaur oJ lwrrrw.f MrI(rlwe; Mroit PrGwwesr Section. Yak U•Mr.Mr Sebol of Medicin.. New Havea. Cewa UNA S. RYAN. f N D_ Srwior SrknNU, Pywkal.o. Cancer Research lae/4 11ne. Mi•wi• Ae.ocWe Professor ~/ Mrlklwr. Uwiverw of Mia.i Sclool of Medicine. Miam~ Fle. B. V. RAMA SASTRY. D.St., t„.D. Professor of rA.nn.cology. Vander- biR Uni.ereity ScAool of Mediciae, Nashville. Tewn. IAKOR SCHMIDT. MD, f/n.D. A} itat.w/ Professor of aiecAem4r.r. Do- •/ab/r of !biogkd Scknns. State U•heteNF of Nsw York at S/owy Broot. Stn.y Brook. OlRALD SHKLAR, DDS., C4.tn A. frurfea Professor of Oral r.rAotorr He.4 D.p..lnlrwt of Or.l Mr/kI we and O.or l.rilofotr. Harvard Sefool ef DeMd Medid•e. nadoa. NATHAN H. SLOANl. 1 w.D. hoJir aer oJ The u•im.ily of Tenneree Ce•1er for ele Heah` Sciewati Ma.rie. LOUIS A. SOLOFF. M D. ffl.ncAr P. Levy Dip' oliAr/ Srr.ke rro/r.wr; rroJrawr e~ Mrlk/we; Dlrerror. Re- urcA LI~iI Laboratory. Temple I)nl- renitr IleahY Sciences Cenur, rfia- de1rMa. THOMAS P. STOSSPL, M.D, CMrJ. Mrllc.i Onrolotf Unit. Meaaetluaetla Oenrral HoepNal. Soaon. nAVO. W tAt ua:t' a et n.rra... .•/1 ..1....~. M..t...l 1 .wa. 14w•.r PROJRCT Tii1s PJidealdo4kd rndies of tl.e Piade\ Tri• RepMr~ A Seretk study of DNA repalr 1• iwbred Mrahe of wliu Ma•d••Irsder i•.wye Mid iw pI/bwarr f ord.alw•: seaelaeS coa.ratent of il.- twunottlobl/li• A as awarYa of eeo- tloolk aa++h I/INraetio•e of Aornlo•ea with oene of IM p.41.owary.aselJar wall laoue.oa of .iooli.e on the reicw of aoetyleldiwe i• tM wwlaw o(aceM• a•d he faylkatio•e e. 1ine fetal powt\ Central wiooti•k retup/ors Oral e•reiroJe•e~ .i/a.d• A and «l- Moiee Ma.r•alia. M easr•• N •)F1Kewe 1!e ~ ieal a•d biolo~ et.dirs TM role of lecilld• etwkwerof acyltr.•a- hnea (LCAT) 1n eRoleMerol u~1 /o and from Meriolrcral /i.wce and Its InodifkMiow by rnolMy LecNhi•:eAokaetol acyl uaemferase a/ter eatay F••ctiond anatomy of Ibe pJmoeary leacroplatc ilhr rok of macroflAaee induced faclon 111 taKrf Imnlurllly w/ i PRINCIPAL INVIrs'i]CATOR OR iN371717i7ON ANDREW M. TOMETSKO. M.D he.- i4n! and Dirrcror of Rrar.rcA. I.Mroe L.Eoratorieti Ltd, Rocleater, N.Y. JAMES TRAVIS. PrD troJea.ar oj alodlrmiurl. T1r jl•i.enit ofp Georgia. Atbem. GERALD M. TURINO, MD, Professor of Mrliclwr. ColuwOi. University Col- kp of Hlnkiar • SwSco•R New York. EMIL R. UNANl1Q. M.D., lLnYwchrol! Iro a.o. of lw/wr rAolep. Har- .ar. Medkd Scleol,. UNION CARBIDE CORt+ORATION. Nlydear Di.itioq Oak R:dP. Te.s. f ROJECT TITLE hoAlwg .icotir reoepor aW ' of clro.ic oldrrcUv. Ir•d discuss hoteai„k eay.... •.d 1•Y6ion Is er rMynm Chawicd b•de of Uwn datr.ctb. 1d obelrrcti.e 1••R diee•as t fyriop/holoRf of .ae.al ard adhNed anwoFYaRea Rards •f1b chwockrb2dom of a•Ld ii•lation es*o.rr• M.Ip• ..d d.i- anal STBPHfdN P. VATNBR. M.D_ Aancire •ro/rearo/ Ndkiwr. Harvard Medi- al Seloolr New Pft/ad Re•ioe~d h4 a.Me Research Ce.kr. Sowlsboro. Mar aaYllw/e• AuoeiMe M Medkiwr. Peter aal Rri{Raa IloeFil.l, Ror1o•. IRENE Y. rYANQ PwA, Antrr rro jetwr e( /..k and CBwic.i /al/llwol- ep and ltflaaNdop Medial U•i- .a.Mr d South C•roli•,. CMtRMO.. GEORGE WFJNRAUM. h/.D. I(o.rl- • ewrlu. raJrwowr Dww SecHo•. AI- se .i'edkal C..w . Mii•- IAMP.S A. WILL. D.V.M. h.D. Nro- fewr and CA.Ir/r/.•. Department of Yere.t•ry a~~ . University of rVi.= OEooORO1R EMWd31 DD lna. rro/ra.or of rArsiofodlnl ~Aen.Wry. DerarMwwr yN.Nrifio• and Foo! Science. Mara- ch.reua LAMwe ef Tecb.olop. Caal- bridp. KOII YOSHINAOA, Pw.D. A.aocl.te rro/raw of Anaanr. 4jaraar of Nanw. RyroJrcNlow wwd Repro/rc- H.e tJofedy. Harvard Medical SeYool. Sorlon. (Now al Ae Center for Po~ INioe Researcb. NICHD. Nationd ln- aNe+1a of HeahR Rethnda. MJ) N ~-Md.e.d reiea oao•rS +w~ . P.rMca/b of .whiFie forr .f .aw li.ee sakrwo•ad eybclrora P4lR tapeM// tM eAcie mctiof dpr•~NN~.wo~. M~IW Ma.pion M ic .nd fwKllo.al ooree/Miw off IM t(/D odle ef Va 1••R Vitawti• A •wd~ym•roleiw eynhe.ie M neFir•wry e Lel _iww,• w loeyl Iramfer•.e •eo..1!1e early rr.~r~ ~ -e rr~- er e•eflne fae b~edder caneer fffede of nicoaGr on pretaa.e7 91

r Completed Projects Followint is a Got of the principal iovestisaton, or ioslitutions, of projecb that have been completed prior to the period covered in this Rcport. Several of the individualt named are deceased. The titles and dfilia- tboa listed arc those in effect at the time the work was completed. HYLAN A. BICKERMAN, M.D_ Astlu- SUE sUCKINOHAM, M.D. A..hu.r am Professor o/ Hedicine, ssd AL- t•ro/eswr of Pedialrks, Columbia U.i- VAN L. BARACH. M-D. Consultant venity Collcp of Pfy.icisr isr- iw Mtdiciwe, Columbia University Col- {eoas. New Ywk. lege of P6yskissr k Sur~eoro - Oold- wster Memorial Hospital. New Yort. A. SONIA sU1ST, MD., Amociue M tuoe of Medk4w and rA/sbbfl. B1O-RFSEARCH CONSULTANTS, m.cnitr of Ore{ow Health Scie.a. INC, Cambridge, Mw. Center. Portl.wd. LEO O. ABOOD, /rrt.D., Pro/rasw of RODA A. sARNES, M.D.. PH D.. Pro- B10-RESP.ARCII INSTITUTE INC BENJAMIN BURROWS. M.D. Asss eiochemistr7 awd lraiw ResrrcA. Css. Jtmor (AONiate) of Physlofogr. Colo- , ., CamMidAe Mu. elre Pro(esser of MedldAe. Udvetdtt 1cr tor Breita Research. The Unhrenitr rdo SIMe University. Fort Collins. , . of CskeM Cbleep. of Rocherter Medical Center. Rocbe.• FRED O. ROCK PN D Associate Caw- E M BUTT M Chief rath-I-&& D ter, N. Y. FREDERICK W. BARNF.S. la., M.D.. , . crr ReurcA SckwtW, IiofoAkul Srt . . . . . Los Angeles CowMP Oener.l AssmcWe Professor of aLdlawe, The Now RosweR Park Memorial Ir1iMe Los Anaeks CLARENC.C M. AORFSS. M.D., Aaer /o1w HoCis University Sclwd of , , SOrinaviBe N Y . d.re Cliwkal Pro,essor o~ Abdklwe. Mediciwr, ehirnae. , . . RICHARD U. BYPRRUM, ht.D., hv University of Csliforwie li(edic.l Ce. OUENTHER fiODF N M D As.x/ate (!s.e' of CA.wdur 7, MkalAs. Sta1e ecr, Los Angeles. T. C. BARNES. D.Sc.. Resrrch Sciew- . , ., Professor of MeNcine: Asslstawt Dkec- ~~rsitl. East t.wia{& ANTHONY A. AI taANESQ ht D. Di- rfu. MRedclOtia Stste Hospul, PAils- delpt.l.. rr, General Csinkd Research Cewrer, Tem Us~ersiy Health Sciewoa Cen ~y.~R M. EMILY CAH M.D, rrctor of Laf,oratoritr, The B.rb Re- ~ CAeirnw, Department e/ C wWr7. fsAitil.lio. Ceaer. Wt.ite Plsius, N.Y. CARL O. BECKER. M.D. Associate ~ Regis Cd1eK Weuo~ pftuy. . i HERMAN V. BOENIO, h.D. Head. BRUCE P CAMERON D AtD M ANTIIONY P. AMAROSt. Pw.D. Is .ersHT Medical CotkgeNe. York. DrParrnwnr of Chemistry arad SiocAeer- . . . , . rd Ho l i How Ud N r urrrtor h Olsrrnks and Cynrcologl, iur7, Spindletop Research Ceaer Le:- s .sl Wle, a wre i / f M~ S h The AIEsny Medical College of Union Universiiy, AlEeny, N.Y. , R. FREDERICK eECKER, Pw D.. Asso- inaton, Ky. ciete rro/issor of Awaom7 and Dirrc- len t iward S r L t• f P o ad of Medki.oIs.IM c m or c r. orl o n a ce, JAMES F. SONNER, h.D, Professor i D U l WILLIAM H. CARNCS, M.D, U.Mr- Pro- H. T. ANOPI-AKOS. M D.. PN.D. n uke versity Medica Center. Dur- l.wkwe of C skr of UteJ Coll e of Msdkl.e SsR , (eur of PAysiofotl. Boslon University School of Medicine Boston. A.re, N. C. Totr. P as.de.a q . L.te City. . D. MURRNY ANOEVINfl, M D. l)ni- •erspf of Wisconsin School of Medi- RALPH S. BECKER, PM.D., Professor of CAernisrry. University of Hmulow, mwaoo. WALTER M. BOOKER. Pw.D.. Pro%r aor ond Head. Drp.n menr o/ Phrrwo• cology. Howard U.i.ersAT. Wst.iy- MARCUS N. CARROLL, l.. 1'rD, CAfrl. Di.isbw of Phernm.cefep. The Rrooltdsle Hoyild Cuiur, Meoilm cine, Medlsom. BENJAMIN BPI.L, M.D., Direcror F.mrr- tor. D.C. N. Y. DOMINOO M. AVIADO. M.D. Pro(rs- sam of rAriwacoforf Uni.er.itf of Itus, Normative Aging Srrdy, Veler- .ns AdminiNrstion Ortp.lie+1 Clinic. TOM O. BOWERY. PM.D., Resr.rcA ho%ssor, f*uk/de Resldur l~oro- WILLIAM ALVIN CARTER. MD Assistant rro(essoe of Medk4sr o~ . Phil- Pe.wylv.wie School of Medicine sono.. Norty CaroWr Slae College. M wkrobiallse1. TM lotwpook1s lh4 . sdelphi.. SAMUEL oPLLLT, MD.. Dirrctor, Di- att. ver.Mr School of MediciMS, 1.Nl.we. rfsbw of Crdiology. Philadelp6i. OEOFFRF.Y L. BRINKMAN, M.D.. As- LEOPOLD R. CERECEDO l7t.D h. STEPHEN M. AYRES. M.D.. Di.ecto., Oe.eral Ilo.piul, Philadelp6i s. seciarr Pro(essor of Medkiwe. Wepsu . . /e.br o/ esocAee.6rry owd Nsrr/fi.rt Crdiow/wcowr7 Lloratorr, SsIM Stste University School of Medkiae. University of Puerto Rico School e~ Vincesr's HoyNsl, New York. BARUI SENACERRAP, M.D.. Fa6yaw Detroit. Medicitsk Sea 1ualL hoftssor and Choire..w. Drprtwrrwt of OSCAR 1. l1ALCHUM, Pq D., HasrMp t'ariblop. Harvard Medical School. ROBERT B. BROOKS. PM.D. Associate JACK CHALON, M.D. Assoc/.ti PrSkr Professor of Nedrelwe. Uni.ereily o( bpoe. rro%ssr of rathofoR~, Ud.enNT of ~or ol Anruhes/eloRr. New Yort d- Sowt,er. Csli(orwia School of Mcdi- O.epw Medical SctwW, Porllsrrd. .a*y Medksl Cewler, Ne. Yos\. cine, t.os Aryeks. rro(cssor of JOHN A. BEVAN M.D. , ljniversily of Celifor- Phrmacd.#l r BARBARA s. BROWN. Pn.D. Chief. CHII DREN-3 HOSP(TAL OP LO0 AN- BANO D DERIK B M Professor FRF , ei. School of Medicine. t.oa Angeles. lSrprrimewtaf Psychiatry. Veter.ns Ad- OCLFS, t.os Asyeka ., . . . . and Chairn.ew. Department of ParAe- minislrstion Hospits/. Sepuhreda. Cal. No1ogy. TM Johns Hooins University School of Hygiene and Public HcJtb, BUDHOF.V BNAOAT, PN.D, Professor SeiM Louis University of Physiology R AYMOND R. BROWN. 1'n.D. ho/es- SANFORD CHODOdH, M.D. Asdwiu Professor of M.dkbe, Tahs Uslr.a Sshimore. . School of Medicine. SI. Louis. aor o( Clinical Owcolop. Uni.eniry of W i l di M di sity School of Mcdkine, Boror< iscons n Me , cal Schoo a son. Pro/.r NAITPR M CHOPRA ht D M D IFFORD !AR(31!R CI Robert A Dlrhlon M 1) CF SARP BIANCIFIORI . . , . , . . . P/rodrr Pro/rtuw of Phyrlol- Hrwr . . . Univcniq of of Cancrr Research 1OSEF BRO7l!K, Pst D., Professor and sr ot CAanisrry. North Carolina Aa- ' orr, Iter.^srd Mcdicei Sc". Boston. . Perusis, Pcrugis, Ilsly. CAairman, Department of Psychology. t.eAigh Universily, Bethlehem. Pa. rkuhurel and Technical StNe U.her- sily, Orccmboro. Pro(essor of Pathology Cornell Uai- 92 93

WILUAM O. CLARK, Pr.D, Director. rsychoPhrws.colot? Reurch [Ai1or.- ary. Velerene Adrninielr.tion Ho.pilel, Sepulveda. C.1. HANS T. CLARKE. DSC. P. Jeseo. of eiochnnl.rry. Columbia University College of Myslciens & Surrons. New York. JAY D. COFFMAN. M.D, Seedew Heod, Peripheral vescdr Dqornwent. University Hos;iul, doslo.. ALLEN s. COHEN. M D Pn.D. A,- socire Professor ol AfeJkine. Cthie% r.d.w/wy Stctinw. Temple Us>tvsnity Scie.oes (:wer, Plail.delplei{a DANIEL COHEN. D.V.M, M.P.H., A. siu.wt Professor o Yettrlw..y Ep{- Iritrloloty r+I rrfllie Headrh. Uni..r- wwy of 'ewytveni. Sebol of Vaw- iwary Mediciwq PlailadelpAi.. JULIUS H. COMROF_ 1.. M.D., Drte- sor. C.rliorescrlor Reurch Iwst4rt. Universily of California Medical Cen- kr. San Fr.ncisco. DEAN M. CONNORS. M D.. Associate OJnclar, De~..re:e.t ol V~•rry Medicine. St. M.ry i Ho.pitd. Madisom. Wi.. PH1UP COOPER, M.D. Cflwk.l rro- /essor of Srsery o.d Director. Srg4 ed Laboratory of Cellrl.r Physiology . AR+ert Eiatei. College of Medicino af~ YeJlv. U.iversit'•• Chie/, Sw Serrke, Vqer.s Adwiaistr.lion or Dile{L Tfe Rrou, N. Y. JOHN e. CRAIOHEAD. M.D.. rro%r ,or of rrholon U.iversity of Ver- wwM College d Medidoe. lurlinpon. ROBERT L. CRAIN. PN.D.. Assistant Professor of Socldoty. University of C1iuRo. Cricap. T. TIMOTHY CROCKER. M.D. rroJes- sr of Medk(we. University of C.M- (orni. Collede of Medicine. Irrir. CARROLL.E. CROSS. M.D.. ADoci'ote rroleswr of M.Iktnt and Human Physldoly: Direcror Section of rd- nro+.ry Mrdklne. (jnivers3ly of Cdi- (orwi. Scltool of Medicine. Davis. C.ECII. E. CROSS. Research Deprrwunt. Si. losepA Hosppui. Burbank. Cal. ALBERT DAMON. M.D. Pti D, Lec- trer on AwrhropOloPy: Research A,so- ciott In Medical Anrheoplotl. Pe.- body Musetans, Hervard Usaver.ity, CambridK Mus. THOMAS R. DAWBER. M.D. Associate rro%sror of Medicine. Soeto. Udvee- dly Seiooi of Medidr, Boston. R. F. DAWSON. MID, rroleseae o/ Rot- ~ly, ColtrsEi. University. New York. JOHN P. DELANEY. M.D, PN.D., A,- ,ncMa rro/essor of Sr~s , University ef Miresol.. Miewmyolu? ANDREW i DllNER, Pr D., Erec- rhe reychs•Reserch. TLN Age Cen- Mc J New EsRlur/, loe, Boston. EDWARD F. DOMINO. M.D, rrole,- aer of rhrwsocology. U.iversily of MkYR.n, Ar Arbor. RALPH L. DORFMAN. PM.D., Director of Iabr«orie,, Worcester Fouadalios for EaPerieewld Biology. Slrewsbury, Mw. H. FRED DOWNEY PN D. Assistant rroleasr of rhysiolo~~. Univereil' ol Tea.s HeaU\ Seienoe Cenmer at Ddlas: Director. Crlior.scrl.r Research. Car- dioPdmon.ry Isailwe, Metbodisl Hos pild of Dallas. Dallas. JAMES 1. DYAR. Pw.D., Assistant rro- /essor o/ l/ioloty, dellarmine College. Louisvi{e, Ky. RICHARD H. EARLE, M.D.. Chief. rrbwowory F.uctlon L.lororory: A.- ,lswnc ho%,sor of h/edkine. U.iver- sity of Chk.Ra Clitado. JOHN W. ECKST8IN M.D., Asd,t.wr Aolessr of l.rer no~ Afe:kine. Slsle University of Iowa College of Medi- cise, low. City. BERTRAM EICHEI., D.D.S. Director. butkwe of SeowwoloNc.l Research. Scienca Resources Fou.dation. Wa1er- lowss. Mar. HYMAN ENOELOERO. M D.. Arrewd- p~hd, Los Asrida.ue of Ls banoe Hoe- CARLTON K. ERICKSON. PH D, Ae- ,oclre Prolessor of rh.r....coro`~ .wd Toskolojy. The niversily of Kan.ea Scbool of Pb.rm.ey. Lewrence. 94 I HENRY 1. ESlER, PrD, Re,trrA /w- rwrwoloPW, Meso. Rese..ca IrtN.w Woreesler, Mw. JOHN R. ESTERLY. M.D, A,sacilre Professor o/ Pathology. Uulv.r.ity o( Chicago Ptilaler Scleooi of Mediei... C6icago. HUGH E. EVANS. M.D., Dbecto., Dr- p.twrenr of redl.nic,, )ewisR Hospi- tal .ed Medical Ce.1er of RrooklyR, Rrookly., N. Y. 11ANS 1. EYSENCK, Pw.D., DSc rro /essor o/ r,~cholop. Inslialo o~ PsP. ~ . Univer.ily of Lowao~ Lor~ HANS L. PAl K. PN.D., Al/rwn As,.cF ote Professor of Paholo". U.ivertily M~S ~ ~ Cdu«.i. Sc.oel et AP4dm DANA L. FARNSWORTH, MD., WewrP X. Olirer rro%.ro. Director. Udver.lty Herrh Servkea, Harvard U.iver.ity. G.beiya, M.e.. GAD FEINSTEIN. Pw.D. Senior Leee tswr /n >fibchr . The George S. Wi.e Center o! Life Sciancim Td Atk U.iversity, Td Aviv. Iersid. FRANK C. FERGUSON. 1a. M.D. Ch.i.w..w, DeFranewr o/ rArn..col- o`y. Tb Albany Medical Coqqe of I).io. University. AR+.oy, N.Y. THEODORE N. FINLEY. M.D. Dii.ea toe, rdrwow.ry Restrch LdorrraeP. MowM 7io. Hos0eL S.n Francisco. WI~LUE~ M 1. FISHREI - Saudi A!M Clelcaller IIe.M1, Cfk of~ EDWIN R. FISHER, M.D. Director of L.boraork; Sb.dydde l~oyitel• rra- tessr of 'rhole~~. Universfiy of ri1//brrdR Scbd of Medicioe. Pilw eutr... RUSSELL S. PISHER. M.D., IlnivenU~ o/ Maryland Sclool of Medkine, Ral- timore. ' S. L. FREEDLANDER. M.D.. Director 9 C.wcer Research. Moernt Ziow Hoe- i.i .ed Medical Center. San Fr.w- CiKo. FREDERIC A. FRP-N(:11. AB.. Direc- ror of C.ncer Chemotherapy Research MouM 2ion Hospild and Mediej Center. San Fr.ocisco. JACK PREUND, M.D. Asdr..t rro- Ieua of rhrw.ocolosy. MeDco) C,o1- kp of Viqidk Riel.od. GILBERT H. PRIEDELL, MD_, CW/ oJ rothoJosr. St. V lfeyiutho.a w~monNr. Mw. GARY D. FRIEDMAN. M.D. Aaslr.r Dinewr, De~rt~w~.r e/ Itferad Merh- o/. Rear..cM, KaYer .Po.d.,tyoe 4 nead LMilaw O.W.4 Cal. H. HUGH FUDENBERQ, M.D.,e( H.ler +r of Medklnt Usifver.iry (aII- farni. Medkal ~ener, S.. Fr..eisn: Professor of lt.ctr/olo ? and bn.o- ~ f. Udver.lty of C.Jtfw.i~ Ruf. ARTIIUR FURST. PuD. D/ncw, i+ t+Mrweq Cbwsk.( .lotop. Utlivarslty eif 3.m ltr..d.oo, 3.. Francisco. MURRAY t. GARDNER. MD Adr Nw rro/essor of rrholop ~1.Ler• dlr of Soabee. C.ufor.i. 3ewa ot Medicin». Lo. A.ilelea GEORGE O. OEY, M.D. Dr.ne., rir ntr-ilowell C.wee. Research 1.o1.ro- rory: Associate rro/tssr o/ Jrpry, M~, 1 ~ .i.u.lt~ adsd~. THOMAS M. OOCKI: M.D. AswrWe hole,wr of r.erenH.e Medicine w1 Cawwwwky Health. Seto. H.M Cob ef Medki.. ..d De.liwry. leney . N. 1. DAVID M. oOLDLNRERq, lcD. M.D, Associate Professor of r.t4d• K7. Temple Ud..e.Ny HuMi> Sd- e.oes Center. Pfil.delRii.. PAUL QOLDHASER, D.D.S., Awcd.n Professor of %rblowtoWy. Harvard Sellool of Den/d Meditine, Bore.. LEONIDE GOLDSTEIN. D3c., Ass dere Peole,we o~ r, cr AWiy~ew de for Menlal Ile.h ie.oe~Se Medicine A De.tis/ry of New hney. Rwdere Medical Sc•ool, PiaMaway. IRA OORE M D.. rro/es.or o/ r.rbf- op. RoNon Utlver.it~ School of M.M- aoe: Chief of [~+oerr, Serrke. VNen.e Adw.inistrNio. Ho.pital, WeM RoaMury, Mw. GERTRUDE Y. OOTi3C//A1.1_ Pr D. Assistowr Professor of Iiochrw.letry. Columbia University College of Psyd- ciasn S Suqeoow New York. 95

A. CLARK ORIFFIN, PM.D, Hrad, Department o/ d/ochrmtarry, M. D. Ar.detso. Ho.pitd and Tu.ar Inri- Iwe, UsiversMy of Teaa. Medical Center, Housto.. ARTHUR L OROSS, MS, Senlor I1o- cAawW, SowkweM Researc! Inaituta, San AMonio, Tea. MORTON 1. OROSSMAN, M.D.. Pw.D. w»ociw Cfinkrf lroIes.or oJ Nrdi- c/nr. Universlty of CJifornia Medieal Cenler, Los As>selea. CARL C. ORUHIIT, M.D. PwD. Aa •scbte !n Ph> > and rhrm.col- ally. UdversMy of nw.ylvan/n Orad- uate School of Medicine. Pbiladelpila. )O6EPH 1. OUARNERI, Pw.D, wttrnd- iwg llfkrol)iofogiat; Dirrctr. Afkrol4 olorl Labratr/ri, Long Island lew- Ish-Hillslde Medical Center. Queens Nosp/al Center ARWation, lamakrt, FRA YN K E. OUTHRIE, PtrD, rro,r.- sor of Enromolot r. Norts Carolina Slate College. RakilA. H. R. HAAG. M.D. rrofes,r J rhr- nr.cofolf. Medical Collega ol irtinia. RicAmond. F. l. IIADDY, M.D. PN O, Iro/rtwr .nd ChaJrman, Drtaartmrnt of lhjrb ology, UniversNy of Oklahoma Medical Center. Oklahorea City. JOSEPH 11. HAFKENSCHIEL, M.D. Director. CrdioPulmonry l/nlt, Tla L.nkenau Hosp/al- Associate h. Alyd!- eine. University of iennsylvani. School of Medicine. Ptiladelp6ia. BERNARD HANES. PwD, Professor of Health Science. California Stale Univer- eity, Nonlridte. RICHARD 1. NAVF.L• M.DY Assistant Professor of Afrbriwr. Unwersilr of C.irfornia Medical Cenler, San Fran- tisco. IIERBFRT R. HAWTHORNE. M D, ChaLm.ra. Dr~.rrment of Srr~rry, Univenw~ of Penro~l.ani. OraduaN School of Medkine. PtAadelOAia. IOIIN A. HAYES. M.D, Associate rathdogl,r, Mallory Inqitule of Pa- tM,loar. Buwon City Hosptal. Boslon. f I ARK W. IIFATl1. M D. Professor of Alr.lrrnnr and /hrr, ror of Hratrh Srrr- l.rr. luhs tlnrversury. Mrdfurd. M.u. PAULINE HEI7PR, PrD, Research Associate M Cytofotl and Cyrochrmia- ~. S.o Fr..ciw (trtilule of Medical Sckttoa~ Sam Francisco. LAWRENCB L HESTER. 1r . MD, Professor and Ch.irwr.n. Dr)v. rmrr.r ./ Obtrsks and Gynecolo~, Medical CuOq" of South Carolina. Clarkslo.. PJM CURTiS HOFF. M.D. PM.D., lro%aar awf CM/rnwn, Dirl.ion o Colkp/ lrfehfrrk Rearrrh. Medieal of Vkrsi.M, RicYwo.d. RUSSELL L. HOLMAN, M.D., I.oulei- ... S1aM UnivenllP School of Med1- clnti New Orkana. OLE A. HOLTERMANN MD., Re- itarch ScirnrW. LoflrnJ LboratorP. Ur.«.Rr of Nare Dame. Notre Dame. Ind. FREDDY HOMBUROER M.D., rrrrf- drwt and Dlrector. Biof~esearc! In.U- lute, l.e, Carnlri~e, Mass. ROBERT W. HULL, Pw.D, Professor V IioloPkot Sciences. Florida Slale ni.ersity, Tdia6anea. RONALD R. HUTCHINSON, Pw D.. Re- .rarch D4rctr. Foundation for Behav- iord Raeuc4 Autusta, Mich. IIT RESEARCH INSTTTUTE• Chicago. GEORGE JACOBSON. M.D.. Professor and Hrnd. Department of Radldogy. Usiversit~ of Southern California School ol Medicine, Loe Angeles. JERRY HART JACOBSON. M.D., DI- rrctr, Dir4lon of Electrotrhyalology. New York Eye .ud Ear InBrmary, New Yort. JULIUS H. JACOBSON 11. M.D, Arro- ciare Iro%ur of Surgery and Director J o SwPicat Research. Univeruty of ernroM College of Medicine. Bar- linpon. MURRAY E. )ARVIK, Ptt.D., Arwciate rro/tasr of rhrmecofop, Albert Ein- stein Cdk{e of Medicine of Yeshiva Uni.e»NP, 'I~e Brona, N. Y. OSWALD R. IONES. M.D.. St. Luka'e IbsOilal, New York. ANnREW A. KANDUTSCII. PM.D., Sros Scientist. Tbe Jackson l.aborn- Iory. Bar Ilartwr, Me. 96 I ARNOLD R. KAPLAN. PND Dbec- tr, L.borar.ry of Mrddcrf ~iewrlk; Ckveland Psychiatric Inr Y/~ NoepiW, Cle.el.nd. ATTALLAH KAPPAl, M.D. rrojeasar end Senior rhliJden, Tre ~ocbfellv U.ivaatr, New York. IIRATCH KASPARIAN, M.D. Asdar- ant Dirtctr, C~d/oreicnfar (aboro. trP; inrtructor in Medider, Hakr- nurr Medical Coilep and Hnqitat Philadelphia. ELIHU KAT~ Pn.D. Associate rro[r~.~- wr of Sociology. tlnlveniy e+f (14 cyq Chicago. SHIRLEY L. KAUFPMAN, MD, h~r /rur o/ ratJblot7, StN. UM.a~y of Ne. York Dowtwate Medical Ce.. 1er, Brootly.. ANCEL KEYS, Pu.D, Directar, Lobor.- /arp of rhyi/e/od/r~.f H7t~. Univer- aq' of Minnesota Scaod af Publk HeaN4 Min.e.polia. IOSEPH ~. KIRSNER, M.D. !re/ra.er lklow S~cbol of M~ ~ Chicago PETER H. KNAPP. M.D. Research lro/rtsor of ri~cM.n1. SoMos Uni- versity Sc~oof ol Medid.R, beeto.. KENNETH P. KNU07SON, M.D UttF veniq af Washington School et ~ledi• ei.e, Seattle. ALVIN 1. KOSAK. PN.D. Associate ho/n.or of Chend.rrP, New York Universily, New York. . ROBERT A. KUHN. M.D., A.soc/ate rro/r.sor, Dirislow of Newow.FerP, New Jersey Slale College o( Medrcine, lersey City. ST1O KUI.I.ANDF.R, M D., Professor and Chaomaw, Department of O6.rec- rki and Gynecology. Uni.ersily o/ Lund. Lund, Sweden. ~ MARVIN KUSCIINER• M.D. New York University Medical CeMer. New York. C1IARLFS W. IABP.Lt.P.. PM D.. Anirt- ant Professor of Enr/ronmrn/al /l)- sirwe. Icllerson Medical Cdlqe, Phil.- delp/ua. 97 AARON l. LADMAN. Pw.D, !re/ane and Chdnw.n. Department of Awa- aw1, TM UnlveraMy of New Mulo. SAod o[ MeAfcie.~ ARnsVrerqr., THOMAS C. LAIPPLY, MD, l1~/e. •or of larAol N.rthwsf.rn U.1- •er.itr Mdio.t~ CtriuR.~ MICHAEL & LAMM, MD, lro/iasew of cal P. . New Yert. U~~+aNf M.~ PAUL S. LARSON Pr.D N..~ w of VirBi.iRicdoo~.d.• ~Nkai t~ ROOER K. LARSON. M.D. Chief Ak/ of Metfdnr Fceas Cow He*Nelf Prwq CL1, GUSTAVE A. LAURENZL M: . CMr/ 4 Medk/ne, St. V lle+R.~inoau wr«ee.w, Mar. JOSEPH M. LAUWERYN. MD, S PM.D., Professor OrNnrUu and CMk- n..R, DeFrtwvnr o/ %t rnd ~kr ~ wwwerwP. FiH ~ op, Katoliete U~.i.e~r.iter it111e Le..eR LwsR adgirtw. EDWARD LEETE, Prt.D. D.Sc he/ir tr o M ( _C~iwtstrf~. Univenily J Mirw• RICHARD A. LERNER. M.D Associate Mewrber, Scripps Cliak .wJ Research Fou.A.Boq La 1o11R Cd. CECILE LEUCHTENBEROE ftD, He.st, Department oJ C Swir Institute for Ea Caw ar ReseucA, Lwnwn., SwiuMl.oL AVERILL A. I.IEBOW M.D. CAd, wwn, Dy.rrmrwr of ~'arhototr, Y.k Univenity School of Medk1.A New Haven, Con.. F..4fF.N O. 11NDSP.TH, M D. PwD, !1. IosepA'a Hospital Researcd Laboratory. St. Paul. Minn. ROBERT H. LINNELL, Pw D Arxl- ah Professor of Ch.wdary, dnlvarelty of Vernronl, fiurlinpon. HERBERT L. LOMBARD. M.D.. M.P.H., Asif:.tr, C.Arrr Rrwrch ta- nirMtr, New Entiand Deaconer Hot o11a1. Boston.

J. P. LONO. rfr.D. Professor of rAr- rw.colagy, SrMe Unlvereill of low. Cdkp of Medicine. tow. City. CLAYTON O. LOOSLI. hr.D, M.D H.ulwp Professor of Mr/klns .w3 r.rAolo~~, Univer.il~ of Sourher. CaU- /omi. Sc1od o[ Medicink Los Ayekt. DONALD R. LOURIA. M.D.. A..eeMt Professor of MedcMs. Caaetl Ud.rr- ei1y Medical Cdkp, New York. KENNETH MERRILL LYNCH. M.D Sc.D. LL.D., Professor Emre/hu .~ Pathology and Chancellor. eJk./M Cdk@t of South C.rdi.r< CherleMOte. INES MANDI. Iu D.. Auluowr rro%r .e.e/ NorAtiwlwry. Cdwnbi. Upiver- .My Cdkte o[ Physicians A Swseo.L New Y [or JOHN H. MANHOI D. 1... D.MD r.o/saaor a! dLenor. DsprNnswr oj r.rAo!o~~ .w/ Or.f Dlo~nosir. New ler.er Cdk~e d Mediciwe .nd Dcw /iMry, Jerse~ CAf. DAVID E. MANN. Pe D. AuocWe Professor .f rArwr.rology. Temofe Ueivereity School of Plermecr. nrl.- dc)pisie. FRANK ARTHUR MANNINO. M D. Auiu.1u Professor of O6rrrnk. •wd Olnttofe~~, Wonxn'e Ho.p~el. Loe A~ye/d Ca.n1~/Univerw~ d Sowberr. C.tiforrr. Medka) Cawer. l.m Anleke. JOHN P. MANOl, M D., Instructor M Yirowy .nd 1.crreiofors. Mediul CdkRe of Sow! Carolina. Cl.rkero.. CHRISTOPHER M. MARTIN, M.D AuW.wr Professor o/ Mslkds .w~ Dbsrtr. Di./dos of fn/tcNoru Dia soau, Scion H.R College of Medicine. lereer Cilr. MASON RESEARCH WorceMer. Mer. HENRY C. McVilh h., M.D.. Acting Hsd, D.p.rriweal of ruAolorr. Loai- .i.r JuM U.iver.ily Sclooi r,f Medi- ci.., New Orle.r. HENRY D. McINTO6H. M.D., rro/r.- ae+ o/ Medicine and Dirscro.. Crdio- r..ewfr Iabarrory Duk. UniverekL Medical Ceeter. Drrh.w. N. C PORDe A. McIVER. M.D.. A'uocl.rs rro/sssr of Pathology. Medit.i Col- IeM 4( South Cardi.K CYerkelo.. EDWARD MeKER M.D., Professor and AcNnt CAdrwrw,Dry..rwrrws of r~rM1.gy. Medipi (' ge of South Crdi.K Ciwk.row. KELLY T. McKEQ M.D. A..oci.re rro/s.wr of Mdk4.e. Medkal Cd- Iq. ef Sow` C.rolir, Ci.rkro.. VK.'TOR A. McKUliCIC, M.D_ rro/s. aor of Melk4v. The Jdme Hoptiu. Un/.er.itr Sc>od of Medicine. (lehi- .nr.. ROSS L MCLEAN. M.D.. Associate Professor of Afsdlkk iws. Elworr Ueives- .Nf School of Medicin., At1.M.. WILLIAM P. McNARY. 1.. Pu.D., Ar .oci.rs Professor of Awetowry. Souom University School of Medicine. Souon. NeAL L. McNtVEN. Pr.D.. The Woe- « uer Four,d.rion ror Eaperimenid Biology. Shrew.Eury, Mass. JULIA MEYER. h1.D., Associate rro- ,sraw of Or.1 r.r . University of 1/1iwM Cotlcde of Dea ry. Chicago. DOV MICHAELI. PM.D, A..isranr rro- (!>sr of /iochsrnlm) and Sw~rr~ Uwhrenilr o( California Scbol o~ MedkLr, Sam Francisco. INSTiIIIit; DONAI D 1. MASSARO. M D., Auod- re Professor of M.Jichs. George NruYnpo. Urtlvtnily School of Medicine. Waihyloq D. C. CHARLES McARTHUR. M D, rry- cAdosirf. Usisrrily Health Srrrker. Harverd Univer.irr. Cembridge. Meu_ CHARI ES s Mct'ANTS. IN D. Ar.e- rl.rr Iru(rsu.r o/ t.•Jr, N.woA Cero• 11ne Creie l'olkge ti.lrwl .rf Apoeul• wie, Ra1r.gA BERNARD 1. MILLER. M.D.. A..luenr rro/saeer of Awrowry. leAet+oe Medi- e.l Colkm Kil.delEA1.. JAMES 0. MILLER. M.D. M.D, rio- /sue. ./ Psychiatry and r,ycAolo~y; Direcrr Mtnaf Health RrsercA fn- .Nnrs, bavenNr of Michigan. Aw Arbor. CHARLES MITTMAN. M D.. Director. Drprnrwnr of RrrOirerory Dioreirs. CNy of Ilope N.rionel MedH el Cen• Icr. Duerre. Cel. 98 I HUGH MONTQOMERY, MD. Aaro- ci.rs rro%uor of Msdiciiw Udvenhy of Pena. Ivani. School o~ Medkior~ Mil.del~i.. P. O'/. MONTOOMERY 1.. M.D rrofeawr of rrtrotot>, ~l a~ Texas SowhweMert f~(edkd~3cRod, D.11.e. OEOROE E. MOORI~ M.D. PMD Di- rrc.or. Rorwetl ttirl Meweri.l ~rMl- /we, Bu/alq N. Y. KENNETH M. MOSER. M.D. Aabe" Professor of MslkMr. Oeoqefowr U.iver.N~ Medical 3cbol. W..YM- 1oRD.C. IIURLEY LEE MOTLEY. M.D, hoJs} .r Mrlk/ws and DMscre., CrNs; Rs .ror~ [.letora.~~Uni.esN~ aP So.r~er~ Cdiforwi. Scfod af Medi- ciwe, k.o. ArRela. EDMOND ANTHONY MURPHY MD. 3c.D. Associate hojraw .j /bwNulu and MdkMs, T'M Idwr ~~ u~ eMy School ef Medl- QYILLIAM S. MURRAY. Sc.D. Sewbr Sa/ Sckwtlu, The l.ckw. L.bors- 1orl. Bar Hrbor, Me. RICHARD L. NAlYQ M.D. rr./e»w and cweww, narrr ot r.diol- Cd-. k~ Hershey. OEORO 11. NEURATH. Pw.D., Mkro• w.yrkd L.lor.arE. H.aMrR, Wed Ckfff"Wf• ALBERT H. NIDEN, M.D., rro/rt.or •~ Mslkiwe, Drew !"orllpr.drNe Medk.) School .wd Uni.etny o1 Southern C.titorei.; Chief. rnlrwowry Dlrase Section. M.nis Lwber King Herpiul, Lo. Angeke. OAK RIDOE NATIONAI. LA/ORA- TORY. O.k Ridge. Tera. , DONALD M. rACE. M D.. rrolea.or of Physiology ond Director. iwuir.us lZ Cerr.rr.r RersrcA. University of bruk., Lincoln. ALlERT B. rALMP.R. Pw.D.. Aad.r.wr rro/r.>a+r of Pathology. Univereily of Toledo. Toledo. O. ROSE MARIE PANOBORN, MS. A. afuw Food Tschnolor4r .w/ l.Mr.r, Ds of Food Science ..d T.cA- wo~y U.ia venil7 of Cditorda, D..Y. JOHN W. P./ARK~ER, M.D. Aa..elrr 3~m Gli%~r.i. - !cRo d l of dffie. Los Angeles. MARY Si1rARNR PAR3HLEY, PaD.. Assistant M(ta+r N Aw.aw' ir O~• Mstriu rrI Qywseo/osy Cd..ki. U.F.enryCdMRx et Qt,la{cwe a 8.r- poa N.+r aetY EDWARD W. PELIKAN, M.D., Choi, ~ EaDq. ~ T of rA..+w.e~f U.HweMr Schad .f M~t /oMea MALC.'OI.M C. PIKB, ftr.D,A./..soy of CewwwW7 M.dki.r .wd reMer. .ka, U.ive~.ai!~ef SarUiw. C.Rfrd. seRool of Medid.., l.a. Aryelr. OTAKAR 1. POI.LAK, MD. ArD, Eatawbs Dtncror, Doree Medial R.. .e.rcf CepNr, l.e., Dwer. DeL M No.we.~llolnrt, U ,~.t C. M. POMERAT, DPM•.DM ., Dr~ N I/olotk.l RsxreA, Pwdew. Poriir lioo (or Medipl Reeeat~ Per/e.rt, C.1. S N. PRADHAN. M.D Pn.D. M/so- ,ne r~i.nwae/o~l, ilow.rd U.i.et- eMr ~oMe.f et Medki.k W.rRMDeq D.G ~ M~r.D~,._Mfs. H.R r.TrT ~THO~ +vCaNe of p o/ SowL C.rdiD.R, Ne/c.rClrYww PROCESS A INS11lUMlNT CORlO• RATION, Srooly.. N. Y. MARTiN S. PROTZPI, DD.! CAi. , Cil .~aNOral . tNN ir~•~~ WALTER REDLSCI/. M.D Ar.el.w rn/saar M CfMkd MeAkl.e. New York Uwieer.ilr School of MeMeMS .nd NYU Research Service. Qail.Ner Maaerid Hoyir.l, New Yak. TIMOTHY 1. REOAN. M.D.. Professor V Medk4we: Dirscroq Dl.l.l.w o~ rdiow.cdr Dl....er. Cdk .'M Medicine sad Deai.lry of New Jersey. New Jersey Medied School Newark. 99

WILLIAM REOELSON, M.D. Professor JOHN A. ROSECRANS. Ih1.D., Asso• CARL C. SFI.TZER PH D Honorr and CAoYnNn. DrParInKM of Medical ciare lro/rtsr o/ tl.rrnrolugr. Medi- , . .. y RrsercA Auociae Ikabody Mtrsetre OncologT, Medical Cdkge of Virliwia, d olk~a of VirRLhk VYRiou Cam- , , Harvard UnivcrsilY Cantbridge Mns RicAmosd. weall~ Universily, Ric,atond. . . . LU('1O SEVERI, M.D Dktner and LYNNE M. RPJD, M D. Wo1brA -ro. JOHN R. ROWLANDS` PN.D., Stt/ . Dron. Iwulrrre of Anorowy af rofAol- !e ~o rorAololf. Hanard Medical Scknrlsr, Southwest Researc! Onsuiauwe, rf. DivYion of Caneer Resesrelr Se C~rrmtw, Dehrtnwnt SM AMoeio, Tea. O v nieersitr of PenKia Peeryia Italy of rswho%q, CRildra's Hospital „ . . Medical Ceaer, Boston. RENIAMIN A. RUBIN. M.D.. Aulsanr CHARLES R. SHAW, M.D. CAk/, See- Pn/trto- ~/ Pr-Ik Health. Baylor riar of A/rdknl Gtnetka M D Aa- HOBART A. REIMANN, M.D. lro(er~ Uaissrdly Colkie of Medicine, Ilar. , . . derww Hospital and Trrwor loslillsle; sor of Mrdklwr, HaMeenM Medical .aw. no/eur of tlolop 71e Univenilr College and HoM11d, PhRadelpYia. , of reaas at Herwoq Ht>relow. RONALD P. RUBIN, M D., 1'rofrssr ROLLAND C. REYNOI.DS, M.D. Ar rArrwocoloar Medical c:dkp of CHARLES E SHERWOOD M D As- Nsrrnf rrolrawr of rrAoloq. UaJvei• . t i rsllaili. RicVnoww. . , . r1slw/ hoftswr of R~~ r Udrsr- sNr ofTesas Sowllweuere Medlul sMf of Roc-esNr ScRool o/ ~bdklM Scbd, Dallas, HENRY 1. RUSSP.K, M.D. lrrsldrnl, and De«hlrr, Rochester. N. Y. TY Rtrm! FornddioR, inc. Suncn VICTOR RICHARDS. M.D. CAIr/ of lrlawd, N. Y. SHOII SHIBATA, M.D. hM.D. hio/rr Swferl. Presbfutias Medical CeaMr, sr rliornrec .Unl.ersi,r of San Francisco. W. C. RUSSELI M.D Udvwxsily of :~ Haw School of Howolnln. Teus Medical Zeaer l/orna, ARNIS RICHTERS, IhM.D, Associate . . DAVID L. SIMON MD fsunrnor Is rrofr,sor of %rAology. Uoivenilf of WAYNE L. RYAN PM.D l-ol'ruor of , fwrtrwd AfeJklnr. CineIl)rlNi General SoaRern California School of Mediciwe, , - diodwnWry University of Ncbraske Hispiul, CMeirwarL Loe AncOeks. . Colkp of Medicine. OmaAa. ERIK SKINHOI M D CAk/ Dirw- WILLIS H. RIESEN, Pw D, Srnlor !fo- PETER F SALISBURY M D Pw D , . . , ntwr of Newefop ttkpalivft Hoyi. cArntiu, LiJe Sclencrs Dlvlslow, IIT . , . _ . . Hraf /ntrnsirr Trrtenunf Centrr , Wt Co/ed,qe. Reuarch InrMuu, Chicago. , , Saint Joseph HosPiul, Burbank. Cal. . THEODORE A SLOTKIN Pw D Aa- DANIEL R. RIFKIN. M.D. Asilsran- PAUL SALTMAN MD Assistant rroa . . . sisfMr Professor of r~rnwtrofop, rro/rswr of CArntknl BiulorT. The , . rssor of eioeArnrfsh and Nwritlon Duke Unirerritr Medkal CeMe- Our. Rockefeller Uaiver+uy, New York. , •iversilr of Sow~er. California . bow, N.C. ScRod o[ Medicine Los An eles R. H. RIODON, M D. Professor of re- . g . GEORGE W. SMETiflRS~ MD.. Aar MoWY UaiversMr of Teaa Medical k ULRICH H D4ee- SCHAEPPI M D rlne in ro- s, NortMeslarl ua1- srant: OalveMon. . . ., . 1or of Ntwo/Grnrrofory, Mason Re- vertwy MedicalcRoel, Chicago. seveA Iswilwe Woreesler Ma» SYDNEY C. RITiENBERO, 1'N.D, , , . GENE M. SMITH, M.D. Adsratw M- rro/essor of ll.cteriulolr. University IOROEN U SCHI EOEL M D PN D feswr of Psychology. Harvard Medieal of Sw1Aern California. Los Angeles. . . , . ., . ., ho%ssor and CAtbnwn. DrNrnnrnf ~ Atneus Oennal Hoyi- BENSON B ROE MD Aswcbrt Pra- o / Sr-j+r~Talane Universilr School ., . , Jtuor of Sw~ rryr; [:ANI Crdfoc Sr- M MNk New Orkana. LUCILE SMITH, Pu.D.. ho/inor e( Sr , Uwlversilr of Cd1/o-nh School ~edkMse Ss>w Franchos. ALVIN R. SCHMIDT. PM.D., Dirrcrr _ IlerAtnd>.rf, Darlwoww Melkal Schod Hanover N H , , , . . .4 Cr.wnsrfint, Tof1e UnivenNr, Med• lO6F.t•H It. ROOFRS. M.D. Holy Nanw fwd, Mass. SHELDON C. SOMMERS, M.D.. Dbrte- of lews Iloyild, Oaddeq Ala. rr of LrAorarlrs Letwa HiR Noa- , ISAAC SCHOUR. D.DS., hMD, D.Sc., al• Cl/nkel Professor of latMolop, ROBERT C. ROSAN M.D.. Associate Dean. University of 1111nuis College of olwnsia Univetsil College otPUyd- . hn/rssw of %rAolos and rr/itrrks. Dnwistrr, Chicago. eians A Suraeor~ ILtr York. - Sr. l.auis University ~Iwd of Medi- eine; Associate ratAoloslu Cardinal SCRIPPS CLINIC AND RESEARCH ERNEST SONDHEIMER fn D Auo- . Oknnon Memorial Hospital for C41- dren, SI Ltwtis. FOUNDATION, 1.• Jolla. Cal. MA1IR1CF S. SFOAL, M D. Chnirn! ('11AR1 PS 1 R(KP rM D. Cl,..,. n..rr. rr../r...r of AfrJutwr, Iufts Ilnwrt. r.., utr 1aMwd of Mrdatnc. Or. Vru.6wt 1.4w,a,.t,a.rr 4 4.pr1rM Nn,,,rwt ,./ b.Aelur.nw Tl~rropr. Hus( 1,m, M...~..w I..n t.1y 11u+p.td, Butlun , . , ciart Professor of elocAends/ry. Cal- ktie of Foreeuy, Su1e UnieenMf of New York. Syracvse. T. M. SONNI?BORN, PND, D1nln- rwlshrd Servicr Professor of Zoology. Indiana Untversilr. Rloominllow. SAM SOROP, PM.D., Ht.4 Dtrrnws.r Of Mor7otwo/acrlr CAtndspry, The ~~: for Cancer Reaearcl. !Y- SOUTHWEST RESEARCH INSTI- TUTE. San Aronlo, Taa. DAVID M. SrA1N, M.D Dbecw Dt- ~d ~. AofotT. ~. Rrea~dsla ALEXANDER SPOCK, M.D., Aulnwor rrs/tasr Pc+dW-k; Duke Usliver• skr Cewlee, Drrwll% N. C. FREDERICK J. STARIM M.D Prof... aw of Nutrition Harvard llaiveri~ Sclwol el Public IbaN14 RoMoa C. HAROLD STEFPBB, M.D, Dticw- of Lo-rrra-ka, Methodist Hoylta(, Mew/bM. JACK P. STRONG, M.D. AaseeMM hojrarr of Pa-A~/o~f,1.orYMoa SIaM University SeRoel 07 Medkira, Now Orleans. MARION R. SULZt1LRQER, MD. M- It.N. .sI CA.trnw, De'.n,wrw of Derwrofoq and SlphfologyNew York UnivenMl.ttsRrew Ma/le~l Cwr ur. New York. RENATO TAOIURI, h.D, AsrcMN powessair Of of Setareiw~ea RiA~ Hanrrd Unlversily, Rael". MARC D. THAM6S„ M.D.. Sewlr Qa urrA /Fellorr, Mayo CIWa and Pw& dsriorl. Reclerer. MMn. CAROLINE BEDELL THOMAS, M.D. le\aa HeNl~nf UaJNr711f~SAYel at Medkiwe. DaRMwor~. JEROME P. THOMAS, h1.D. ta>.r of Sawl/rr Ebrtrlnt, U~ of California, fler~eky. JAMES H. P. TOMAN, /w.D., ftie/tr se-owd CWrwsow, Deparww wf of ritr r. Chicago Medieal Selroai, l*- aNrM or Medical Resnrcl< Chicago. JANET TRAVELL., M.D., AaaoeMw Professor of CUnkd IArwsooelsp, Corr.eR UniversNT Medical Cdkre, New York. LIE SHA TSAI, Pn D., ReurcA Assr clLse In r«Aoloq. Yak University School of Medicine. New Ilavew, Cone. 101

D. M. TURNER. M.D., Head. Deprt- inenl Dr.R Meradollsnw and S(o- cArwr , Harletow Laboratories E.- rope, LW , Itarropte. Yoekshire, Eft- laad. UNIVERSITY OF SAN FRANCISCO. San Fr.neaca UNIVERSITY OP SOUTHERN CALI- FORNIA, Loe AMycks. ELLIOTS. VESELL. M.D, rro~esaao..n/ CAoirws.n. Deportment of rll.rws.co{- otr. hMsrlvnia Stae U.ieersity Cd- kse of Mcdidne. Milton S. Hershey Me diul CecMer. Ikrafey. BRANISLAV VID1C, D3., hofesasr of Anatomy. Oeorgctovf Uniteniy ScAoob of Medicine awA De.tWy. WaAitrgton, D.C. ROMEO A. VIDONE, M.D. A.aorWe hoftssor of raaholojl. Yak Vsi.er- aitF ScRool of Medic.ne. New Ha.e., Con.. PETER K. VOOT, rtt D. f•rofewor of RfwroNofogy. Uni.ersitr o1 WasAiq- lo. Sdod of Medicine. Seaitk. E. D. WARNER. M.D.. lrofessor of r.- rAolos'. StNe U.i.ersity of Iowa Col- ktte d M0iciwe, Iew. Citr. SHIELDS WARREN. M D., Direcror of L.boratorks. Cancer RrnwcA lnn!- rrwe. New Ey/a.d Deaoo.en Hay{- 1d. hl.owoa. YASUSHI WATANABE. h1D~ Arrr elre MernM. Tu Wix.c iMtiNtN ef AwNaef and BiobR1. KiladelpRi.. BARBARA K. WATSON, ht.D. AsrW- ont Rorwriolo0n. MassacMrseus Oew- .r.l HoqMal• RixarcA Aasocine in ~.cr.rinfofy ~ond ln.wrrwolop, Har• .ard Medical Scfool, Boetow. LEE W. WATTENRERO. M.D. /ro/cs- eor of /auAolory. Universitf of Mln• ne.ola Medieal Sciiool. Minneapolis. /OHN 3. WAUGH, Tu.D., rrof.ssor of p% Cherni.ny. MssaeRusetN Instilwe ed N TecAnoloSr. Cambrdge. r B RICHARD 1. WECHSI FR. M D., Cfin- m kal rlYrrwl..rlrt, Mume6ore I/osptal Imhlute of ReseacA. PituhursA. JOHN V. WEIL, M.D., AsrLrant P.o- /euor of Mtdicbsr. University of Colo- rado Medical Center. Dee.cr. A. WPJNSTOCK, hM.D. Research a/o- eAensW, /Jf* Sckncrs Di.irion, IIT Reaearch laditwq CAieaRo. RUSSELL W. WELLER. M.D. rauAol- oe!rf Mewsorid Hopit.l of Ct+eaer CSw~y, Wed Crerer, Pa. A. STANLEY WELTMAN. hr.D., Auo- e/Ma Professor of rlwnwcolo y and Rese.rcA, Rroodh. College of har- .rcr, Brootyr; N. Y. SIMON H. WENDER. M D., Research lr eu.r of IiocAemiarry. Uni.ersitf e( Waww Norman. DUANE O. WEN2EL, M.D., Professor and CAairww, Dyartnwnr of rh.rer.- c y o.I Tock y TM University of Karwas School ~.rwacr, Lasr- reao.. THOMAS C. WESTFALL. M.D., ProIa- aor of Pharmacology. University of Vir- &1. Sclool of MedkMre, Cl+asloues- .Yl.. FREDERICK E. WHISKIN. M.D. C.M., Dirtcror, Division of Health awd f er- .oo.li.y Epr'GMbre, Tin Age Center of New Eylaa4 l.e. BoMO.. ROOERA ~1- WILLIAMS. M.D. rr..feasor of C : Dnecrer, Clyton Forw- d.rius /iocAnwkof lnaitaxe, T1e U.i- .enity oI TeaaR Awti.. DANIEL H. WISEMAN, M.D., A.si.t- aM Professor of lelwrici. Uni.enit~ of BaMberw Calitornh• Clwldreri s Di• •MioR Loa Angeles CowNr General Hospiul, Los Anaekt. 1. EDWIN WOOD. M.D., lo.tru.•tor Iw MeJkMc, Ilm1ow University Sc4ool of Mcdkine, Sostow. SUMNER WOOD. IR., M.D.. Assistant hofefwr of ratlbiory, The /obns Hopiies University Scrool of Mcdi- cine. Baltimore. lOIIN P. WYATT, M.D. Irofr.+nr of rutAolop. Saint l.ouis l/ni.crsity Scbool of Mediciee, St. Louis. INDEX OF PR1N(3PAL INVESTIGATORS Abood, L.. O.. 40.41 Ace[o, M. D., 59 Arnott, M. S., 9, 11 Benedict. W. F..9 Bing, R.1., 32.33.31,36.37.38 Buid. A. S.. 78 Canlrell, E. T.,14 Castro, A., 76.77 Cfiak+n, l., 32 Cross, C. E.. 22 Crumpacker, D. W.. I 1 Fsantan, W. B.. 43 Friednsan, Q. D., 79 (kokaa, M. C.. 60, 61, 68 Oiekn,l. H., I S akkA. a.1., 6t (ioWstein, L. 39 Oorrod, l. W.,16,17,18 Oueti., M. R.. 10, 63.64 Queloo,ll. L.,12, 13 H.R, L M., 48.49 H.nro.A, M., 27.46 Heaoowitz, H. B., 69.71 1o.cAitn, H. L., 20 Iaw(, A.. 28.29.30.70.73 lsuto, W.1., 62 Kleinertwtu, J.. 26 I Kari, R.1-, 9 KuRander, S.. 79 LajtA., A., 39.40 Lt.y, l. A.. 70 Lieeer,l. E., 30 l.yncA, H. T.. 20.21 MaNin, R. R.. 11 McClearw, O. E., 42, 43 MitcAeM, J. A., 44 Murad, F.. 57, 58 Oeacb, F.,14,15,16 h*a, K.. 22 Pol.t, l. M., 74 RaMwb, l., s0 Reytwld., H. Y., 23 Rran, U. s.. 24, 2S, 26 Saqry, B. V. R.. 36 Sdbrtidt, l.. 47 Sb.we, N. H.,19 Solo/. L A.. 33 Stouel,T. P., 39,67 Tn.ic,l.,49,50.31,52,33,34,33 Turkno, O. M.' 27.22 Uaawn, B. R:, 73, 74, 73 Vutw,S. F., 34, 33 WiR,1.A»31,72 YosAinp, K.. 43 . v IV 103 m 102 ~

INDQ Abood. L 0.. 40.41 Acelo. M. D., 39 belkr, D.1., 73 sing, R.1., 3), 36, 3i SraugAkr,1. M.. 37 Sroa.cAi, E. A., 59 Suist, A. S.. 78 Card,1. P.. 44 Car*, H.. 7) Caslipo, M.1., 55 Cas1% A.. 76, 77 Ca1o.L 1. E.1r., 66 CAaloe,1., 32 Chalurvedi, A K.,16 Croa, C. E., 22 Daeuai, L A., 17 Del Mar, E. C.. 61 Ewnr. W. R..4) Farr, A. O., 74. 75 Freedmaw, H.1. 12, 1) Friedmaw, O. D. 79 Oayk, T. M., 64. 67 Geokas, M. C.. 60. 62 OeOwer.1.1., 48 Olaq, H. R., 16 Okiclk 0.1.. 68 Oortod. l. W.,17, 11 Orees, M. R., 22 Oraer, C. A.. 43 OuaiL M. R., 10, 63.64 Ourloo, H. L,12 Ha616Mow. D. L., 26 Hall, t., M., 49 Hama/ti M» 27,16 Herw..dez-VasOuez, A.. 31 HencowUs, H. s., 71 Hiuiwti C. E., 65 IIoMnberg, R. W.. 66 Hosaker, C. .., 65 11oM1. s.. 47 lanoR, A., 28, 39 loMsow, D.. S l )wko. W.1., 62 Kyrio, l.. 90 Kowi, R. E., 9 L.cto, A. 0.. 33 1 OF 3EWIOR Aifl'IIOR3 LarAn, C., 61 Levy,1. A.. 70 L'w,W.1.,19 Loren:,1., 14 Lynch. H. T.. 20.21 Marchand. C. M-F., 47 Martodam, R. R., 30 Marwy.~~a, Y.. 37 Ma1Ae~.., N. R., 32 McLenwre, T. L, 9 Mowcybu.,1. H., 63 Mwad. F., !t Nakajirna, K.. SO Neksew,1. M.. 39 f.+, V.. 16 higen, K., 22 rakucA. E, 20 hrMae, K.1., 69 hnson, r-H., 79 Rawp, V.. 26 ~ Reyaolda, H. Y., 23 Rodrituet,1. R., 27 Rurkerford,l. D.. 34 Ryan,1. W-, 24, 23 Ryan, U. S., 24 Sastry, s. V. R., 36 Sershee, H.. 39.40 SAurin, S. a., 67 Sloane, N. H.. 19 Smodgras, D. R.,11 Swkely, l. R., 63, 66 Tepoer,1. M., 42 Treaoa, C. M., 72 Travit,l., 49, S 1, 53 Tlner, H. W.,14 Va. Ca.dort,1.,11 VaedeeAerg. S. O.. I 1 Valner, S. F.. 35 Virca, O. D., 54 WeiiMw. R., 32.35 WAarbn,l., 74 WAile, R., 30.70 WilNams, S. S.. I 1 Wong. D. M., 71 Yeatb, L IN., 63 Yo.binap, K.. 45 104 . I . .