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~ IMPIRIM ~vo . .. . •` 'L'f,. ~ tW a . , ~.. . _..~.... 1976 REI'ORT o/ TIIE COUNCIL EOR TOBACCO RESEARCII-U.S.A., lae. TNE COUNCIL FOR TOBACf.O Rt:St:ARfal-U.!{.A., Inc. 110 Ea.l S9t1.91ree1, New Yorh, N.Y. 10022 44

I SCIENTIFIC ADVISORY IIOARI) to The Council for Tobacco Rcsearch-U.S A.. Inc. as of December 31. 1976 SHELDON C. SOMMERS. M.D.. Chairman Director of Laborator(es. Lenoi 11ill Hospital Clinical Pro/rssor of Pathology College of Physicians B Surgeons of Columbia University New York, New York RICHARD M. BING, M.D. Director of Cardiology and Intramural Medkinr Hunlington Memorial Hospital, Pasadena, California Professor o/ Medicine University of Southern California School of Medicine Loa Angeles. California JOSEPH D. FELDMAN. M D. Head. De artment of Immunopathology Scripps CPnic and Research Foundation La lolla, California WILLIAM U. GARDNER. PH.D. Scientific Diractor, The Council for Tobacco Research-U.S.A., Inc. E. K. Hunt Pro/essor of Anatonry (emeritus) Yak University School of Medicine New Naven, Connecticut ROBERT 1. IIUEBNER. M.D. Chief. Laboratory of RNA Tumor Viruses National Canoer inslitute Bethesda. Marrlaod I I I LFON O. JACOBSON. M.D. Director. The Franklin McLean Memorial Research Institute Regenstein Pro/essor of Biological Sciences University of Chka" Chkato, Illinois u. AVERILL A. LIEBOW. M.D. 1 u+ ~ Pro/rssor of Pathology (emeritus) University of California School of Medicine Saa Diego. C.lifornia , t HENRY T. LYNCN, M.D. i Pro/risor and Chairman Department of Preventive Medicine and Public ltealth Creighton University School of Medicine Omaho, Nebraska HANS MEIER. D.V.M.. Dr. Med. Vct., M.R.S.11. senior Stae Scientist The Jackson Laboratory Bar Harbor, Maine I.EF W. WATTENBERG, M.D. Professor of Pathology IhCartment of Laboratory Medicine and Patnolo6Y University of Minnesota Medical School Minneapolis, Minnesota JOHN P. WYATT, M.D. Director Tobacco and Health Research Institute University of Kentucky Lexington. Kentucky 9dd.tlfe St.ff .f The Council WILLIAM U. GARDNER. Prt.D. Sckatl/ic Director ROBERT C. HOCKETT, PH.D. Reaerrb Director JOHN 11. KREISHER. PN.D. VINCENT F. LISANTI. D.M.D. Associate Research Director Associate Research Director DAVID STONE. PH.D. Astorlare Research Director

0 CONTENTS Studies Related to ('ardiovascular !)iccascs and Function .... 5 Abstracts o( Reports . . , . ... - . , , , , . . . . 14 Cancer-Rclated Studies . . . . . . . . , . , , 14 The Respiratory System . . . . . . . . . , , , 29 Heart and Circulation . . . . . . . . . . . . . 41 Ncuropharmacolo6y and Physiology . . . . . . . . 54 Immunology and Adaptive Mechanisms - . . . . . - 61 Epidcmiolo6y . . . . . . . . . . . . . . . 64 Active Projects . . . . . . . . . . . . . . . . . 67 Compkted Pro jects . . . . . . . . . . . . . . . . 75 IndeR of Principal Investigators . . . . . . . . . . . . 85 Ldex of Senior Authors . . . . . . . . . . . . . . 86 t Studies Related to Cardiovascular Diseases and Function Previous annual reports have presented the general plan and rationak of Council-sponsored sludies of earcirapnesis and the etiology of chronic pul- rrN.nary disnrders. Their purpose was to provide a framework in which Ihe rekvance of individual eontribulions would he more easily apparent. In the present issue, we describe similarly some of our appruaches to study of cardiovascular diseases and function. Cardiovascular Dl.e..es Among Ihe many disorders of the cardiovascular system, those deriving from progressive alherosckrosis rank On1 as causes of disability and death in the United Srates. These include heart attacks (myocardiat in/arclion), angina, slroke, arlerial blockages in the Iirnbs, and some cases of congestive hear/ failure. Hypertension is believed no1 only to accelerate alherosclerosis, but alut to precipitate acute events In damaged circulatory syslems. Atherosclerosis is, lherefore, a principal focus of the present discussion Epidemiology o/ At6ero.clero.ii-Related Di.eo.e. Numerous epidemiological studies during the lasl 20 years have southt possible "eauses" (primary or contributory) of the alherosckrosis-relaled dis- eases. These studies often surnmariud their findings in terms of "risk /aclors " The "risk faclor" is essenliaMy a statistical concept based upon mathematical relationships without necessarily any known or established mechanism by which it might contribute 1o etiolM. Among the many twch "risk faelora" frequently reported are hypertensinn, elevated serum choleslerol, diabetes or a prediabelic dialhesis, cillareue smoking. personality type, Inadequate physical aelivity. and emotional stress. (]enetic predisposition (e.g., hyperlipoproleinemia, hcxmaysteinemia) is another such /aclor, both In its own tight and as a conbibut.x to most ur pi>sibly aN of the olhen. Biochemisls, physiologists, pharmaaologists, psychuk.gisls, and wtKo scienlists have thus been challenged to /11) the gaps and learn whether. how. to what eatent and in what kinds of persons each factor might aclually he operative. The stimulus to eoniec/ure, speculation and hypothesis has Accn strong, and many scientists have apparently seited the occasion lo lest Ihcu personal hunctxs in the hope of a hxky strikc. This was entirely Icgilimatr, but has tended Io produce a weller of (ra`menlary, confusing and inconclusive observations subjected to speculative projections. Clarifkaiion may come as cotxrent, integrative I"ia r:metile that can accommodate all the findinp that prove vdid. S

I Cigarette Smoking as a "Risk Factor" The identification of a"risk factor" can be no more reliahle than the stalislics on which il is based. Smr+ken selccl themselves from among the Seneral population on a basis or bases that are little understood Is q legitimate to c.onspare them with nonsmoker controls as animal esperimcnlers compare their test animals with genetically identical htter-malcs? Tobacco use has many variations. both qualitative and quantitative. To what ealent does voluntary adoption of any particular behavicral pattern select out a sub-population with innNe characteristics or life-styles that are linked to cardiovascular disease swceptihilily? There are many evidences (rons studies of man that genetic predisposition plays aa important part in delermining which individuals are poter tial victims of aUetosckrosis-relaled diseases. In several types ot rather e.lreree suscepli- bilily, the patterns of inheritance have been well established. Review of the "tisk factors" listed above indicates that most of them are already knowa to be inlhrenced, it not determined, by heredity. Studia of sponuneous alcohol cornumption by inbred mice have showa subslanli.l straia differences in appNite, in behavioral responses and in mela- bolism. Analogous mane Mudies of spontaneous nicotine and tobacco amoke Intake have now been inaugurated by a Council grant to learn whether similar genetic variations occar. l'aiw Studies /deetkal Iwins provide the human coumerpart of animals of inbred strain. S/udies of all like-seaed Iwins in Sweden and Finland are being assisted by Tle Couneil in an eRorl to determine broadly the relative inffuences of heredity versus envirronmenl upon the incidence of atherosckrosis-bsed cardiovaseular diseases. A key slratesy Is to compare the incidence of such disorders in smoking and non.moking iden/ical Iwins, including symplomalology during life and, eventuafly, age al death. the primary cause of decease and posl•mortem evaha- lioo of vascular pathology. Non-idenlical, like-sesed twin siblings serve as owMro4. Whik the incidence of discordance in smoking practices is low among IdeNical twiru, an observation that in itself evidences a slrong role of heredi- tvy influences (even among Iwins reared aparl) in determining Initiation and maintenance of snaking, the numbers of discordanta are Sreat enough to accrve b iacrnsingly significant levels In substantial twin populations followed over a Mrllcienl petiod of tLne. These studies have now been eaended beyond Identical twins to other rdalives of various defined degrees of consanguinitr. such as h.lf-aibs and Ihe oRap.ing of twins. 'I1se.w lirneecwssuming iwvesliplinns will require a eonsiderahk induclion perlod before comprehensive new reports emerele. They may evenlually provide more direct evidence whether cigarette smoking per re is a truly si`niAeanl "ri.k factor" in these and perhaps son.e other diseases. They eannN, however, be erpected to add greatly to elucidation of palhotenie processes or to provide 6 9 methods useful in reducing risks due to biochemical aberratinn% imparted by the genes from ancestors. For Ihis, other types of rcaearch are rcquired Cholesterol Metabolism Another prevakM "risk factor" in epidemiological studies has been "ele vated serum choleUerd." Contradictory findings over the years suggest that this concept was an oversirnplificalion, especially since many victims of heart attacks had no hislory of high ehokNerol levels. That cholesterol is implicalcd In a/herosekrosis has Seen assurned because mature arterial plaques contain the steroid both free vtd in combinations. Neverlhekss, long lerm adminis- Iration of drugs that lavet serum ehoksterol has not been as generally elfeclive In arresting or reversinZ the process in man as had been hoped. Accumulating infetmalion about the states of combination of cholesterol in blood is directing aueetion 1o the several c-hole.terol<ontaining combina- lions, classified as ehy:onskrons, very low density lipoproteins (VL1)1.), low density lipoproleins (LOI.) and high density lipoproteins (11D1.). The t Uls are the major carriers of hlood chokslerd and there are a mrmher of eaperi- menlal as well as epiderniologieal /Mrdings that an elevated serum level of VLDI. is correlated with progression of alherosckrosis, whereas ekvaled Hl)1. is a favorable indicdion. Thus certain "lipoprotein proliks° are now considered by many to he better indicators of susceptibility /o atherosclerosis than elevated total chrd- esterol. Certain of these prollks, associated with very high susceptibility to disease, have been showw to be lIeneticaM)f dNermined. tfarring some unforeseeable empirieal discovery (which does occur in medical research and is a perennial hope). the rational route toward effective control is through systematic study of the palboWneais of alherosckrosis, at biochemical and physiob&al kveb. This obviously must include a heltcr understanding of the regulalion of synthesis, lransprxl, tunctiun. and elimin ation of cholesterol and of the aberrations in disease in the hope of altering these by targeted treNmenl. The Council is presently reviewing the mosl promising u.ncepls and kads that now esisl in this eompka field with the intent of increasing its suppoft of basic study of athererosckrosis. The inlent is not only 1o assist these im- porlanl devek+prnenls, but also 10 seek more directly relevant assay systerns for assessing the possible effects of cigarette smoke inhalation. The task Is additionally diflleuH beeause the atherosckrotic disnrders appear to be peculiarly human ones with few eounlerparts among animals under natural eorditions. liighly contrived manipulations to produce animal "oounlerpart:' tend 1o diminish probability of relevance to hrrman esperience Iluman studies have been impeded by the lack of non-interventiwns tech- wiques for assessing iniliNiow and prolresswn of the atherogencsis processes in the vasculalure of man. New eaperimental techniques tor visualizing the condition of the arterial walls, iecluding scintillation photography, appear ar ahow promise for the future. T

In Vitro Studies o/ Arteriei and Vein. Meanwhile, in vitro techniques provide one method of sluJying synthesis .rd uptake of lipids and cholesterol by human arteries and veins, both "normal" and atherosckrolic, in direct comparison with those of .nimais. Though in virro conditions cannot be made to duplicate the in vivo situation perfeclly. they may provide useful guides to in vivo research. Such studies have already been asade with CTR support as described in the Council's Annual Reports for 1972-1973. 11 has been reported (and is reiterated in a current review) that human .Iherosckrotic and nornNl coronary arteries as well as sapheswus veins, perfused under puls.lik .rler.l pressures M(h human plasma containing labeled cholesterol and acetale, do not synthe- site cholesterol from aeetale and produce only small .mounts of cholesterol eqers. Choksterd is laken up in Identical amoums by norm.l and diseased vessels .nd Ihis uptake is increaaed at higher pulse pressures, s.hkh seems eonsiMant with the reputed effects of hypertension. Labeled acetnle is Incor- porated In1o various lipids. both normal and diseased coronary rlerks and saphenous veins synthesize free /atly acids, triglycerides and plwspholipids. Addition of nicotine to the perfusion fluid did not influena' cholesterol uplake. Analogous esperinxnls with dog arteries appeared to shown .n In- fluence of nicotine upon choiesterol uptake. If this is confirmed. iI wttlests caution M estrapolations from this species to man (1972 reporl ). Carbon rna+oside In the perfused plasma was reported to euhanee ehol- ealerd uptake by afl arteries in this in vitro system. Several new studies report a dramatic inhibition of cholesterol uptake by both human and animal arlcrks, under these in vitro eoeditions, by 7-keto- ehoiesterof. Living rabbits also showed an inhibition of cholesterol uptake. hul to a much snsalkr eslent. TAe low solubilily of the 7-keto compound imposed teehnk.l problems. Improved techniques may increase the efficiency of the tRect. Meanwhik. the disparity between the in vitro and whole animal te- sponan is another resninder that eatrapolations must be cautious. Other current studies deal with elucidation of possible mechanisms of the Inhibition and with the metabolic /Ne of the injected 7-keto compound ht rabbits. Role o/ Smooth Mu.c/e Cells in Athero.clerotie Plaque. TAe heretofo.e prevaknl theory of alherosckrotic plaque formation postu- lated that the inflhrNion of (atly substances from the bloodstream inlo the arterid wall gives rise to cholesterol deposits that act as an Irritant, causing In/lammation and proliferation of cells by processes akin to ordinary healing. This "itnudalion" concept was eondstenl with the associations of atherosckr- oda with elevated blood chokslerol, high blood pressure and high lat diets. Animals fed large amoun/s of .aturaed f.ts snd cholesterol. sometimes sup- plrmenled with Iw.nmun.l rw other 1reNmenlt, devek.ped (imirms superfki.lly re.emMrns th...r .+.vr.rd in man •1 aa.trqav 7 he.e lesions often regressed In snrm.l• %tr.• rn. drro oo,e.e atr.-/ •n ~ 1.urvarun that stimulated human dretarr wul.r. rn..a..n~ re.u,r.e.rr ..f atr-IrNer,d snd saturated fat eonsump- r Iion. Complicated by difficulties of conlrol, the results in man have been equivocal. Recent electron microscopic studies of human arterial plaques have re- veakd that the major component of authentic playucs is proliferating smtwnh muscle cells like thrne normally composing the center 1 medial ) layer nf arter- ial walls rather than 6brohiasl cells such as pradiferate to heal a wound. l:arly human arterial lesions (streaks) contain Ii111e lipid. sugResling that insudalion is not the prime factor in typical atherosclerosis. Choleslerol .kptnits and cellular debris appear later. Present debate centers on what incites normal smooth muscle cells to migrate from the medial layer and what causes them to proliferate. Presum.bly, damage to the inner surface cell layer of the vessels is implicated and a number of theories as to how this endrHhelial layer may become damaged have been advanced. Many of the plaques generated In animal arteries by n..r-physiological manipulations are reported to be radically differenl in composition Irom those of genuine human atherosclerosis .nd their relevance to the human diseases is therefore questionable. Recognition of the difference has, however, led to reporta that certain animah can develop human-type atherosclerosis under other more appropriate conditions. Snaolh musck eeYs from primate arteries are now being maintained successfully in culture media. Low-density lipoproteins added to the media stimulate them to multiply. FtMher, il has been reported that the ever-present blood plalelels, normally involved in Ihe clotlin6 process in response to injury and in other functions, ahio secrett a substance that strongly stimulates pro- liferalion of Ihew smooth muscle ee11s. Damage to etsdcNhelial and inlimal cell layers of an artery, which ordinarily separate the bhxxl from contact wilh the smooth muscle cells of the medial layer, can bring the.e cells into contact with platelets and presumably Ineite multiplication in the artery wall. Another concept holds that the snsoMh muscle cells mulliply as the consequence of a mut./ion akin to that which transforms other normal cells into malignant ones. This `nsonocbeaP' Iheory, which is supported by suikin6 evidence but is neverthekss in sonse dispute, would suggest quite diRerent mechanisms of action by e.lernd agents than those describcd heretofore Endothelial C.lti and Blood Plateleta A single layer of endotheliat cells eomposes the thin memhrane lining the inner surfaces of arleries, performing an important function in relainrnt the red napuscks while allowing waer and s.ame other suh.taoces to pass throuth. As mentioned, damage to the endaMhchum is heGeved t+r crrntnhua to all three of the snechanisms of atherosclerosis described. Many p.nubtc causes of endothelial damage have been auRRestcd and are being sturlied It is possible that several may be involved. '1 he damaeed enduthelium can repair itself, rather slowly. but if damage is sustained ar repeated uw often. repair may mN he achieved. (.'lots (Ihrombi) forming in an artery are thuuRht tu in/urc Ihe endo thelium by pressure and by impeding the access a/ oxyRen "1 he hl.wxl plNelets have long been- known to play a role in the ctNnplet events that produce 9 0

tbrrxnbosis ie both arteries and veins, and may thus contribute indirectly to endothelial injury. At Ihe aame lime, it has been thought likely that the plate- kts may contribute In some way to maintaining integrity of the endothelium. Recently developed Iechniques for growing human endothelial cells in culture media, a(ler harvest from umbilical cords, have made it possible to study tacion that influence their replication. A Council-sponsored investigator has reported that platelets added to the medium, as well as some individual subslasces secreted by the platekts, will slimulate their growth. It is, therefore, suggested that !n v/.o, platelets may espedite endolhelial repair. !le has also Irolated from normal blood substances that inhibit platelet adhesion and thus itnpede the processes of thrombus formation. The implication is that a delicate balance among opposing (unctions maintains inlegrity under normal conditions. Estensbn of such studies should help delineate the mechanisms of throm- boaM and atherosckrosis. Another inve.titator has used /w vitro techniques to cuilurr, nt heart wNrele atsd epithelioid cetb /o determine the effects of chokslerd, choksterol- e+less and 6-lipoprwein in producing cellular lipid inclusions and in labilizing }yaowera or mitochoe8ria. Lreldhins Cholesterol Acy/ Tr.ns/er.se (LCAT) Leeilhin: cholesterol acyl transferase is an enzyme thought to be respon- dbie for the esteriAcalinn of lipoprsxein cholesterol in plasma. It i! postulated by seret.l inveMitalon to be important in the mechanisms that renave chol- ~MOrol frorn the arterial wall. A number of previous papen, abslracted in these Annual Reports, have reported dinkal studies of 1.('AT activity in the plaswra of animals and maw sder diRerent circumstances to elucidate its clinical significance. Instability of the estrytwe oomplkaled Ihese studies. E.perimentation to delineate the Mtyswe's wsodes of action required its isolation. purification and stabilization so dmt en:ya+e oooeenlrations could be controlled and substrate compositions betw defined. A reoent publication reports that a concentrated effort has achieved the Iadatiow of LCAT in substantial quuuily in a virtually pure state with greatly Irnproved stability. Development of a radioimmunoassay Is reported to be under way. Two publications on applied studies of LCAT have also appeared reoealti. Availability of the purified enzyme and of i1s radioirnmunoassay should sUmulata and facilitate studies of choksterol metabolism and the roks of the se.esd lipoprolcins and contribute to understanding of atherosckro.is. Massbolic Actioities o j Puln.otaary Endot/te/iel Ceils Pive years of research assisled by The Council have produced nunserous reporta on this subietl. including several published during (976 It has been ,howw that the vasnacsivt potypepUdes bradyhinin and angirAensin I are mtla- bolkdly alsned durrnt a single passage IhrrMith the vast pulrm+nary, vascular bed Bradykimw. a s+.b.taece that tends to lower blood pressure, is completely inactivated while ansiotenein 1 is converted to an6iotemin 11. which is a potent hypertensive agent. Bradykinin, under suitable conditions, can inhibit this antiotensin conversion. These relationships suggest a role of the pulmonary circulation in blood pressure regulation. These metabolic changes have been traced to peplide hydrolase enzymes of the lun`, which are not present in the bioodstream but are attached to Ihe luminal surfaces of endothelial eells, especially those of capillaries and venuks The same enzyme inaclivates bradylinin and converts ansiolensin 1. Steps in the hydrolytic process have been described in considerable detail; antiirxiia lo the enzyme were prepared and labeled to provide lools (or research. New functions of bradytinin are now being discovered, including an effect on prostatlandin synlhetase which remains to be esplored in depth It also remains for future studies to Inquire how dysfunclions of these systems may be related to vascular or puhno..ry, diseases. Chronic Smoke Inad.tion by Doss A technique for chronic eaposure of beagle dogs to cigarette smole via tracheostomy was employed by a CourKtil-supported scientist to took for possi- bk effects on clotling snechawisrns and on cardiovascular function. ikspite recognized IimitNiom ol this technique as a nwQel of human practice, rather e.tensive eaperieece wN,h i4 use for other purposes suggested that e.tension of observations to the cardiovascular system might provide some preliminary data and guidelines (or twure .ludies. No dinically evident dise.se was towrd at/er 1s months' eaposure under different dietary regirnes. Some Indications of possible enhancement of coatu• 1alion mechanisms and of relatively minor alterations in function were re• porled. The author described these minutely wilh cautions against transfer to man in view of eaperisuealal limilalioru, pecies dilferenees, and Ihe treater tbrnpkaNy of human e.viroament..' Studies Innotrins Nleotiwe A number of prW Couneil-sponsored studies, mainly using animals and with a variety ot objectives, have invo(ved the administration of nkotine. Eaperience through the yean wuests that comments on sorne of Ihe problems involved In selection of dosages are pertinent lor evaluati.ns of past results and for designing new and better esperimenls. SmoMers receive nicotine by way of the oral cavity and Ihe luns, in irnolf successive doses over a period of several minutes and repealed at variahle intervals. The entire dosage range they etperience is very low in conlras/ to the levels often used by pharmacologisb in e.ploratory studies of mcoNne's pharmacological potentials. The view has been e.pressed that ordinary smut- ing rarely. U ever, produces high enough levels to act upon the sympathetic ganglia, but slimulates only the several special sensory receptors to incite raprd but brief systemic responses of re(ka origin 7hese responses may be quite contradictory and paradoaical in nature, depending on the conditions that ptedomioate at any moment. 11 10

Yet, within the low range of dosages that can he achieved from smokint, there is still very considerable variation Rate of absorption via oral cavity or lung is highly dependent upon the acid-hase balance in Ihe smoke. Maintenance of blood level is strongly aRected by the hydrogen-ion concentration of the bladder contents. The detree of acidity influences nicotine rewrption and is altered by diet and nervous state. Nicotine metabolism is rapid and melabolic rates not only vary markedly among normal humans but are probably in- fiuetsced by duration of smoking esperience, a recognized but little understood phersoexnon. In addition are the variations due to choice of tobacco product used .nd Individual diRerences in puff volume, puff /rerauency, depth of inhalation. duration of snsoke retenlion, etc. Design of animal esperi.xnts intended to mimic human smoking e>t- perience, long or short, must include consideration of .11 the factors enurner- ated and species diflereeces as well. EaperimeMal control or Mandardiralion of all the variables mentioned in any large-acak or /ong-continued human study appears to be virtually im- posibk. However, an eapan.iors of information on the actual ran" or dura- tions of plasma nicoline levels attained by human smokers (and users of other forms of tobacco) under actual conditions of life should be attainable. Recent studi.s by laborious snethods have provided such data for small sumbers of human subjecls. These are a most valuable interim guide to the daign of animal esperimenls However, mass data on large representative populations are needed to de/lne, on a sound atatistical basis, the ranges, peaks and medians of plasma nicotine on a time scak, if important refinements are to be achieved in epidemiological studies of smoking. R.dioinswsunological A..ay o/ Nicotine Sensilive, specific and rapid assays for plasma nicotine and its major metabolites have long been needed. They should be able to he carried out with very small samples of blood. with simple. inespensive and mobile epuipmenl, and by competent technicians without etaraoadinary special Iraining, have long been needed for such purposes. Rad'aimmunoass.ys would appear to have promise of meeting these re- quiremenls and The Council is continuing to support studies in this area. Antibodies sensitive to nicotine have heen obtained and report.d t1973)• Continuing ellorts are now directed toward improving sensitiviq, testing for specificity and simplifying procedures for broad applicalion. Nicotine Bffecfa on Coronary Circulation in (:on.cioua noR• To avoid the eRects of anesthelics on the pharmacobRical r.•sponses to nieoline, a merhrxf was perfected lor imracar.rlid adminidratiun of the alka- loid to conscir.us do`s in doses de.cribed by the investigator as "realistic." A striking increa.e rn cor(Mary hhM.d flow was oh.erved which was (raeed to Iwo separate /actars Tbe marx component was found to he due indirectly to + an increase in the depth of respiration produced by nicotinc. Ihe minor one to a chemorefiea via a different pathway. F.PIDF,AIIOI.OGIC STUDY OF SiU)KIN(: Ct:SSATION A general difflcully in making and interpreting studies of smoking cessa• tion is that those who discontinue smoking are not cMnen at random hul by their own decision, so that the influence of acleclion on comparafahly of the groups has been unknown. Nor has it generally been possible to as.css the influences of concomitant or "eompensalory" changes in life-stylc, such as alleralions in dier, eaereise, use of akohesi or drusa. etc. A Council-spunsored eomparison of continuing snwskers, with smokers who have disconlinued, and with smokers who have stopped /rK a periwd and then resumed, sometimes repeatedly, is under way. In the population under sludy, a large body of data had been collected by uniform methods when all were smokers, before any had Mopped. Hence, some bases may be found lor assessing selection biases that rn.y have occurred in the aubsequent groupings into the calegories mentioecd. 12

j Abstracts of Reports Following are ahsuacls, approved by the aulhon, of reports on new re- search acknowkdgin` support (rom The Councit that have sppeareJ in scientifk }ournals since publication of the 1975 Report. 1 he name of the recipient is in ilalics. ~ The abstracts are grouped under these headingi: 1. Cancer-Rclated Studies. (1. The Respiratory System. 111. Heart and Circolation, IV. Neuropharn-ucolosy and Physiology. V. ImmunolM and Adaptive Mechanisms. VI. Epidemiology. 1. Cancer-Related Studies HYDROCARBON-NITROSAMINE SYNE:R(31SM AS A POSSIBLE AMPLIFYINO FACTOR IN LUNO TUMORIGENESIS BY TOBACCO SMOKE A number of studies have shown that inducers and repressors of microsomal miaed-funetion o.idases can powerfully inlluence the eflecls of chemical ear- einopens. A significant incidence of pulnwmary tumon has been reported in both rats and mice as a result of the simultaneora administration of )-metfiyl- eholanthrene (MC) and dimelhylnilrotamine (1)MN) at levels at which neither compound h carcinogenic in the lung Nittosamines, including DMN, as well as 7,1-bentopyrene and ),1-bentoAuoranthene whoae carcinogenic properties are very similar to thosc of MC, are present in tobacco snwke. Therefore, it is pro- posed that a substantial proportion of the lung tumor incidence of sn.okers is due to synergism between the carcinogenic hydrocarbons and nitrosamines in the smoke, rather than to hydrocarbons alone. Two alternative tnechanisms which may account for this synerlism are considered. Preliminary results, how- ever, support the concept that DMN increases the hydrocarbon epoaide pool by lowering the activity of microsomal eposide hydrases In the lung while substan- tially Increasing the arylhydrocarbon hydrosylase activity. The demonstration of a DMN-hydrocarbon synergism in human lung earcinobenesis could tn- oours{e uneapbred approaches for the development of partial means of pro- Ieclion for smokers, such as the use of DMN-dcmelhylase repressors and in- hibibrs to block the DMN-hydroearbon synergism. It is also suRgr.tcd that dietary nitrosamines or prenitrosamine components, such as nNrites anJ secon- dary amines, may be found to have sorne role in the etiology of lung cancer. Argus, M. F. and Arros. 1. C. Iorrnd of 7Aro.eN,d d/oloffy 36:491-1911, 1976. Other support: National Cancer Institute. From the Seamen's Memorial Research I ahnratory, I1. S. Public Ileallh Service 11n•pital, and the Uepartment of Medicine. lulane University Medical Center, New Orleans 1 SIRU('IURAI. I.IMIIS OF SPF.CH'1('ITY OF MEIIIYI.('ll()LANIHRENE-REPRES.SIBI.E NIfROSAMINI N-VEALKYLAStS. INHIBITION BY ANALO(: SUBSIRAII-s In order to gain sn insight into the structural specificily of durNthyln,tru,.. mine (DMN)-demethylase, a systematic investigation was carrrcJ wn ,ar lhe cnMnparative dealkylation of DMN, higher diatkylnilrosamines and DMN .n., k+ts, as well as on the inhihilion of DMN-demethylatiun by the DMN ,nawgs_ Other e.periments in this framework eapbred the eQec1 of pretreatnknt by 1. mcthyl choianthrene (MC). a potent repressor of DMN-demcthylase, on th..e different dealkylases. Resuhs showed that the dealkylation of dimethyl-. drcthy6 and dipropylnilrosamine by hepatic microsomes of Sprapre-Dawky rats was tepreaseJ by pretreatment of the anirnah with MC. 7his repression prolus. sively decreased with the Increase d.Ikyl chain length. In conlrast trr iti effect on Ihe demethylation of DIKN. In viro phenoharhital induced rathet than re- pressed the deethylation of dielhylnitrosamine (DEN). lhe rates of dcmcthyla lion of the DMN analog subslrales, although low as compared to 1)MN, in. creased with the acyl chain length. These analogs were potent in virro inhibaon of DMN demethylation when tned in combination with DMN as substrates, and the inhibition decreased with the length of the acyt chain. Although the rate of demethylation of inelhylphenylniuosamine was not influenced by M('- pretrestmenl, tine compound was, however, a potenl inhibitor of demelhylalion when used as substrate in combination wilh DMN. Moreover, beyond the ap- parent distinctness of DMNdemelhylase and DI:N-deethylasc there is now in dication that more Ihan one en:yme, having the same substrate specificity but different kinetic and regulatory eharaeleristics, underlie DMN-demethylase ac- tivity. Arcot, /. C. et e/. Zeirscibi/t Jiir KrebiJo.xArnP nnd It`lin/iche OnAotoRie e6:171-at), 1976. Otller ur'prtr National Cancer lmtilwe. From the Seamen's Memorial Research l.aboralory, U. S. Public lieal,h Service Hos pilal, and the Department of Medkine, Tulane University Medical ('enler. New Orleans. DIMETHYt.N11ROSAMINE-DEMETl/Y1.ASli: ABSEN( E 01; INCRI?ASEI) FN7.YME ('ATABO1 ISM AND MI)1 1/1'1 1('I I Y(1F EFFE('fOR SIIES IN REPRI:SSIUN. Hh.MUI•ROII-IN INVUI VI•MI NI Evidence is presented here that Ihe rate ot decay of Jimcthylnirru.anminc (DMN)-dernethylase following pretreatment with )-merhykhol.rnrhrene (Mt ) is no greater than that so be eapetled (rom rnuoul enryme cat.Mrlism '1 hrse results also show that the observed decrease of 1)MN dcmethyl..e V,,,, Iolluw inb MC administration is rat due to increascJ rate of breakdown MN to dc crea.ed Aa, novo synthesis. Other eapcrimenb in this study inJiaare that dr/fcr eet receptor sites are involved in the repressinn of I)MNdemethyla.e by hydru carbons and by phcnobarbilsl (PB), and that a P-ISI) type nuarosnmal cyto chrome is involved in the demethylalinn of UMN. lhe totality of these e. petimental observaliona presents an apparent patadoa which may tic aummar-

! Ized as follows: (1) MC and PB are potent repres.on of the UMPJ-demethyl- ae; (2) MC and PB ars potent inducers of a number of other miaed function osidases as well as of the synthesis of the essential components, cytochromes P-IIe and P-I30, and (3) the MC- and PB repressible enzyme. DMN-de- methylase, is inhibited by carbon nsonoaide, just s are MC- and F B-inducibk mited-/unction osidases. The implications of these findinp are considered. ArBtn, M. F., Arcos, 1. C.. Pastor, K. Ir., Wu, B. C., and Venkaleun, N. CArmko-Siolotiral Inreraniowt 17:127-110, 1976. Other arpporrr National Cancer Institute and 11oRmann-La Rochr. Inc. From the Seamen's Memorial Research 1.•boratory, U. S. Public Ilealih Service Hospilal, and the Department of Medicine, Tularse University Medical Center, New Orkan.. MALIONANT DISEASE AND TRAC/IEOBRONCHIAL EPITHELIAL MULTINUCLEATION Tr•cheobronchial washings of patients with a wide variety of estrathorack malignancies have been shown earlier to contain si6nifleanlly rrsore multinuckated ciliated eells than those of a matched control group without prediagnosed cancer. TAis study, while confirming the original flndinp, also determined sorsse (aetorn which influence multinuclealron One half of the total group of R24 pa- tients h•d malignant dnease and the dhcr hatf, matched by ses. age (decades) and smoking habit but without predngnosed malhgnancies, served as controls. Mullinuckation was found so be 2 01 t,mes more frequent in the cancer pa- lienta Ihan in the controls In pairents with invasive Iunsors welhout known mNaslases, mulUnuckatson was seen four times more (retfuenlly Ihan in the controls. Site of origin, stage of tumor and escessrve smoking habit in control fesna/es Influenced statistical signifkanoe, but smoking habit in maks did not. A ptospetUve study is now being planned in which incidence and degree of tracheobrorschl•I epithellal muhinuckalion will be used in conjunction with biochemical tests for the diagnosis of occult cancer. CA.bw, J. n d. Canrer )7(.):1074-Iet1, 1976. Othar.upp.rtt U. S. Public lledth Service. From the Dep.rlmenls of Anesthesiology and P•tholopy, New York University Scbool of Medicine. New York. ECTOPIC ISOP_N7.YMES: EXPRESSION OF EMBRYONIC OHNES IN NEOPLASIA Awareness of the phenomenon of ectopic polypeplide hormone production by Iumorn hu coincided with the recognition of ectopic i.oenzymes in eaperi- mental rodent sumors and in the serum and tumor tissues of human cancer 16 I palients- Many newly-recognized embryonic protein phenotypes are recavmR wide attention as °markers' o/ malignancy. A number of other properties b4Nh enzymic and nonenzymic are shared by emhryurssc and neopla+trc cells Ihc ever-increasing accurrrilalion of findings of embryonic gene prrKhkla in ncu- platii• gathered from cesearch on isoenzymes, hrxrrwsnes, and protein antigens is csusin` reevaluation of the current viewpoint regarding the nature ol cancer Tbis discussion es•mines the .uthors' recent etperierrces with the carcinopla- cental •nligen. Regan isoenzyme, as a model system /or studying the regulation of embryonic gene espression in cancer cells. the rekvance of the cell cycle, •nd the nature of the 1ene product in membranes. T heir hope was to construct • perspective on the nature of cancer from the point of view ol ectopic iso enzymes. Tlse evidersoo is then ea•mined from the standpoint of a single cenlral question: "Is the sekrai.e activation of embryonic genes a necessary step in neoplastic Iransforma:ionT' Several areas are scrutinired in an attempt to amwer this question. These include the telalionship between embryology and oncology. the possible embryonic origin of vital transforming `encs, normal host genes, and oncoStnie rrseehanisras. It is concluded Ihal: (1) ectupic iso enzymes, prexnt in /umor lissues but not in the tissue of tumor oritin, •re predominantly embryonic in lype; (2) the Regan- and msn Regan iaoenzyroe- producing Ilel.a cells provide a suitable rnodel syslem lor the arK/y nf a varicsy of (aclon which ue involved in 1he etpression of ectopic iwenzymes of the carcinoembryonic c•1egory; and (3) information on cell cycle and hormonal regulation of embryonic gene etpression in cancer cells may contribute lo our understanding of the role o/ this phenomenon in the process of neoplastic transformation which may reflect a d'rsorder of gene regulation with the re- appearance of trophobl•slie properties. Such traits evidenced by lrans/ormm6 cells may be a consequence of whatever onco`enic agent (chemicals, r•diation, .iruses) produces a speeifie kosa of regulatory control of embryonic Senes. FLAman, W. H. and Singer, R. M. ln: Becker, F. F. (ed.): Canrer: Slolosjp o/ Tnnrors! CeQula. Biology anJ Growrh, New York, Plenum Publishing Corporation, 1975, vol. ), chapl. 3, pp. 37-R0. Other support: Nalional Cancer Institute. From Tufts Cancer Research Center and the Deparlnsenl of Pathology. Tufts University School of Mediciae, Boston. A SIMPLN RAD101MMUNOAS.SAY ()F IIIJMAN P1 Al'UN('AI. At-KAI-INE P11()SPHATASE IRE(IAN IS(H?N7-YM1:) USIN(3 SPECIFIC AN IlBODY POLYMERS Eelopic placental alkaline phosphalase ( ReRan isaenzymc ) has been ftMrnd in the sera of cancer p•Iients, reported in variant forms, and cons«fered to strongly reinforce the contemporary view of the biological importance of cm- bryonie gene activation during weoplaslic transformalion /lowever, the ca Iremely minute amount of Regan Isoenzyme in sera has made its detection, pri- marily by enzymoloifieal quanlil•lion, difRcull Now, this paper presents a com- petitive-prdein-bindin6 assay of Repn isocnzynse using a specific polymeriud 17

! antibody to facililate the phase separation step Only a minute quanuly of the polymerized antibody parlicks is required for each assay in admiclure with the specially prepared labeled and unlaheleJ enzyme. By adding a amall amount of uarch-gel particks before low-speed centrdusalion, complete phase separation can be achieved This radioimmunowsay which can delect 0.4 to 0.8 ng en- zyme protein per tube, is comparable to the sensitivity achieved by enzymic as- says. However, radioimmutsoassay is advantageous to the enzymic assay in being dirsct, specific (no interferenee by the nonplacental-lype alknline phos- phatasn) and capable of detecting both catalyticilly active and inactive forms of Ihe enzyme. Native variants of placental-lype alkaline phosphatase, including Regan isoenzyme and Napo isoetszyme, could thus be directly determined in clinical specimens by this procedure. Chang, C-11., Raam, S., Angcllis, D., t)oellgast, O., and Firhman, W. !l. Cancer RctcarcA 11:1706-1712,1973. Other wrpp.rt t National Cancer Institute. From Tufts Cancer Research Center and the Departmenl of Palholagy, Tufts Ueivenity School of Medicine, Boston. CONVENIFNT IMMUNOFIXATION I:LEC/ROPHORESIS ON CELLUWSE ACETAIE MEMBRANE This paper reports an immunofisalion ekclropboresis technique that can he conveniently performed on cellulose acetate membrane (c.a.m.i, that requires only nanopam levels of the sampk and that oAen advantages nol provided by the use of agar gel as the supporting medium. Applying polyspecifk anliserum direetly upon the surface of e a m. alter ekclrnphorelic separation results in the sinwllarKan 8sation o( nwhipk antigens as discrete immunoprecipitin bands, rnther 1h.w the compie: precipitin arcs observed in conventional immunoekctro- phoresis. Although the principle of this technique is similar to that performed on ajar Bel, sning e.a.m. for immunofiaalion ekctropboresis has the following advantages: (1) Because of c.a.m.•s highly porous n.ture, it provides an e.cel- keM nmlecular<iclusion effect and hence a highly satisfactory ekctrophorNic oepanlion of proteins. (2) It greatly reduces the amount of time needed for antibody /lsation and for subsequent washings. (3) The membrane absorbs much kss anliserum non-speci/kalty than agar gel. (4) This technique is suited for e8eetirey and conveniently amplitying the primary precipi/in hands by re- action with a second-antibodytnryme conjugate which can be recovered and reLLsed. These advantages thus have made the e.a.m. a eonvenienl, acceplaAle, supporting medium for imnunoAsalion ekelrophoresis analysis. Chang. C-11. and Inglis, N. R. (FbAman, W. !I.) (-Nnlca (-A/mlh a A, rn 6S 91-97, 1975 Orher srpp.rr N.r..nal ( anact (nswine Ir'.n lu(u 1•,..r Il.war.h I tntcr •,.) the IrcpatInKn1 u/ 1'alhuloey- fulU t'm.cosat "./...I ,d Mrd.,ne Kn~,.n 19 RIiA('l1ON OF CONCANAVALIN A WfIFI AI.KAIINt PHOSPHAIASI-S EXTRAC7ED FROM VARIOl1S HUMAN IISSUE SOUR('ES Abundant evidence indicates that mo.t vcrtebrale alkaline whether originating in normal or malignant cclls, is intimately as.,K„IcJ „,Ih the plasma membrane of the cell in which the enzyme is synthesized ,,,i1 Ih,r the enzyme invariably contains carbohydrates. In order to shed some I,tM un the nature of the carbohydrate mocity assncialeJ with the en:yme, it w„u/J be of interest to know whether there is a spccilic interaction hetween ,U,hnc phosphalase and concanavalin A(con A) of jackhean,  well chata.rcnreJ lectin which has the property of reacting with cell surface suRar re,aloes A simpk and sensilive ekclrophorctic technique on cellulose uctale nwmhr,,,c was selected to drAerentiate the mobrlities of the free enzyme and Ihc cnrynu con A compk., based m Ihe /ac1 that the compkaed form ia retar.kd al the origin of application dut to its state of ag`regalion, while the /ree enzyme has normal mobility. The spccific binding of alkaline phosphalase was then spwlied by con A aAinily chromatography. The results derswrnstatc that there is a spc- cific reaction between con A and a lractiors of alkaline phosphatases eslrackd from various human tissue sources. This phenomenon was al,o ob.erved wtlh highly purified placental alkaline phosphatase with a specific activity of 200 rmok phenol/mg protein/min. In light of the well-estahlrshed apecificily of con A reactivity toward the carbohydrate moeily, the aulhora conclude that the re- actinB human alkaline phosphatases may contain branched pulysaccharrdcs with a terminal o•D-mannopyranose,.-D-ilucopyranose, /) fructofuranose, their gIo- cosides, or sterically related structures. Chan=, C-H., AnReNis, D. and FliAmaw, W. !f. Moleculer a Crllrlr QJocArmbrry 9( I):SS-37, 1975. Other a..pport: National Cat.cer Itatitute. From Tufts Cancer Research Center and the 1)eparlment of PathotoRy, lults University School of Medicine. Rostbn. THE IMMUNE RESPONSE AT THE TUMOR SI11: IN I 1/N(i CARCINOMA In this study, histologic and immunologic mcans were uud to investiealc the local immune response to lung cancer 7u bcern with. searhoos froto iIr cases of different types and gradct of lung caranunm.n were e+.ruiucJ tonn par.tbvely in order /o evaluate the mwpholoRy uf the hwvl rtvttarn, Ibsluw lrve patterns of strumal cellular reaclion, chau.clcri.hc 1411 Jdlcrenr ht.l0luRl: types of lung carcinorea, were recognized lhc ananutt ul acllular tnfilluuun was highest in squamuus cell earcinornas and luwest or n.wteaslcnt in na4 «/1 earcinomas Within the variuus histologic ealerories. the well JdlerenualrJ tumors appeared to be accompanied by more reaclrve cells than rhe pIMMIy a1J /erenliated ones; there was no relatitm helwcen tum..r nctrasts anJ ccllulr infiltration Ihe plasma cells were distinctly asuw•ia1eJ with sqoauunrs cell car csnunus; their number in the stUorna was proportitmale to the dcsree of dd ferentialion and the presence of keratin prt.Jtced by the tumors In the im mnrndo`ic Iesls, eluates with a high eontenl of inuounuAluhuhns wcr- rc .n

! covered from pleural eRusqns and from solid lung carcinomas by dissociation of anliken-antrhody complesn. In indirecl immunofluorescence tests. Ihese preparations reacted positively with lissue cultures and with fresh suspensions of lung carcinoma cells, but not with cultured cells of most nonpulmonary tumors or with cell suspensions of normal adult and feul lung. Although so far only a limited number of casn has been esamined, the results consistently indicate that tumor-reactire antibodies alre present at the tumor site in amounts significantly larger thui in the circulating blood. It also appears that these im- munoglobulins are largely the product of the plasma cells and lymphocytes accumulated in the tunae sttorna. IoecAbn, H. L.. Dorsett, B. H. and Paluch. E. C.nccr )t(6):22%-2)09, 1976. From the Dep.rtments of Palhology, Lenoa Ni11 Hiapital and the ('olkite of Physicians A Surgeons of Columbia Uni.enity, New Yotk. ACTIVATION OF DEVELOPMENTAL GENES IN NEOPLASfIC TRANSFORMATION Activation of embryonic Renes, as evidenced by the detection of their pro- Ir:in prvduc/s, is being recognized as a genuine manifestation of neoplasia. Since twch pco(eins are being identifkd on the basis o/ germ-layer origin and of eatraembryonic liuues, it seems important to fis the stake of developmenl eorrespordin6 so the pattern of e.pression of such proteins in cuscer tissue. From a map of development focused on events beginning with the zygole and ending with the esuhlishment of fetus and placenu, it is pp»sibk to esplain why certain de.eioprnenlal gene products are restricted to fetus or placenta or distributed in both. It also suggests which products ean be espected to occur concordantly in neoplastK transformation and in neoplasia. In the paper pre- sented here. the espressioa of term placenlal, chorionic and amnionic alkaline pboaphatne uoenzytoes in preneoplaslic and neoplastic lung and other caneers Is asNicipded from a preliminary study of epithelial cell sonic estracts from the tracAeobroechial tree of a patient with btonchoRenic canoer. This study ea- plores /kst the onco-0erelopmenlal relationship between chorionic alkaline pAwph.use and chorionie Ronadotropin, both of which are localized in syn- ekiotroplsoNast and eapteoed in choriocarcinoma. TTsen, the tracheobronchial lree n a human mode! of neoplaaic transformation is presented in relation to t!M known lumor and developmental alkaline plw+phalases. and the widely recognized oacofeld proteim are identified with their counterparts and position io eatly derebpment. Suth a developmental perspective may, when It has been fully espiored, lead Io a rational interpretation of the significance o/ espression of any product ot combination of products in neoplastk transformation and In seoplasia. FttAmmn, W. H. C.wre. Re..a.cA )619 pl. 11): )42) )12a. 1916 Olh.r wP/.rtr Nauonal Cancer Institute Pnxn Tuft. ('ancer Research ('enter and the 1?eparlment of Pathology. Tufts University School of Modrerne. Boston. 20 I VIRUS-SPECIhIC NEUTRAI.IZATION BY A Sl)1.Illil 1: NON- IMMUNO(i1.OBUI.IN FACTUR FOUND NAIl1KA1.I.Y IN NORMAL MOUSE SERA Two types of immunologic responses tu enda6cnous (' type vrruscs h.vc been detecled in normal mouse sera: virua- hindmR as dclecteJ by r.JMnur mune precipitalion auays and virus neulrahratiors. Ilowevcr, while mouu rm- munoglobulins in the sera bind to both eculropic and tenoNruprc IX Iraprc) viruses, these same seta neutralize only the X-Iropic virus. Ibis rnconanlcnty was at first considered to be related to dillerences in the a0unny of thc un munogloFwlim. The dala presented here now indicate that the nerdraliirns Ac livily is mw drre to the :Miviral antihodies detecled by raJrumunurNrprecqHia lion. Rather, virus neutralization is assoeiateJ with a factor whch has rwrne o/ the characteristics of ony known class of nunne tnrmuruiglohuhn It is e0nnve against several X-Iropic virus isolates, but rwrt ecutropic vuuscs, anJ requhes contact of the serum with X-Iropic viros. The decrease of this lacwr in mrnne senrm only afler absorplion with X-Itopic, hn mN ecolropic, virus further demonslrates its specificity for this class of enJoserkwn ('•type virus AlthrMrKh prcliminary, these data suggest that binding of the factor lu the vnus occun and is presumably involved in its neutrahtauon. '1 he specificity of this (actur tor X-tropic virus suggests that it represents a newly recognized type of respmse of the host to an endotenous virus. IIs possible rok in the regulation of enJo Senom C-lype .irss.es is considered. l.ery, l. A. et .1. Proceedings o/ the Nationd Academy o/ Sciencn o/ the Unircd Srotes of Anseriea 72(12):3071-3073, 1975. Other ar"ortt National Institutes of Nealth and the National Cancer Insli- tule. From the Department of Medicine and Cancer Research Instilule, University of California, San Franciseo, the Basic Cancer Research Proeram, Frederick Cancer Research Center, Frederick, Md., and the Clinical Research Cenue. Harrow, Middksea, England. FEt11.OEN MICROSPECTROPNOTOMET RY OF ORAL. ('AN('ER ANI) 1.EUKOPLAKIA Narly In the devek+pment of eancer, a cell line emerges whrne nu.kar deosyribonuckie acid (DNA) eonlent remains conslanl throN-;hom the life .d the tumor. The amount of DNA appears to he largely deperaknl on the a.trral chromosorne number, or karyotype, and can thus he diagnostic of drphotJy. aneuploidy or polyploidy. The demonstralion of hnormal cell hnes in in uru cervical carcinoma has permitled the inference that this change uccurs he/ore invasion, while Ihe fact that this characteristic change is ftwnd in cervKal lesions diagnosed as mi{d, moderate or severe dysplasia wrNrW indreale that an Increase in nuclear DNA might occur much earlier than heretofore suspected lhis study attempts to discover whether oral kukoplakias have 1)NA contents similar to oral earcitamas, and to deletmine the value of F"eulken rnicrospeclro- 71

I pholomelry in prcdicling the Iransformation of Icukoplakia into carcinoma. The technique of using Iwo wavekn6ihs was applied to 10 p sections of 33 ksions and showeJ that five out of ten carcinomas and 12 out o1 16 leuko- plakias had drpluiJ cell lines. Instead of Jenxsnatralins a correlation between the degree of atypia and abnormal Feulten-UNA stem lines. however. the re- suits suggest that DNA content increases at a much earlier stage and that seem- ingly innocuous hyperkeratolic Iesiorn, as well as oral kukoplakias, contain amounts of DNA similar to oral carciroTlsu. This lechniraue, therefore, seems to have minimal value as a predicsor of malignant transformation because the aignifkance of a diploid stem Rne would always be uncertain. Doyk, J. L. and MonAold. 1. H.. Jr. Jownd o/ Dental RerrarcA S,(6):1196-1199, 1975. OlJier aarpporf r American Cancer Society. From the Department of General and Oral Palhololly, New Jersey Denlal School. College of Medicine and Dentistry. Jersey City. IDENTICAL GENETIC BASIS FOR LYMPIIOSARCOMA AND NEMOLY7IC ANEMIA IN THE RABBI f In rabbits, both lymphosarcoma and hemolylic anemia which occur in Iwo genetically related strains. WII/) and X/J respectively, are inherited in aulo- somal recessive fashion. Tests for identily of the hemolylic anemia (ha) and lymphos.rcoma (is) genes show that they are both allelic and identical by descent and that the two different conditions result from interaction of these genes with the host jenotype. 1lemolytic anemia is the primary cau.e of death M compound heterozyRotes, lra/!s, with increasing Iympho-prohferaeive disease with age. Ages at death of the animala with histologically csnllrn.ed disease ranged from Ave days to 22.5 months, the average being about 10.5 months. The symbd ho will henceforth be used to represent the gene (or eilhr.r disorder. Foa, R. R. and Mrlrr, H. The Jo.rrwd o/ Hrrediry 67:99-102, 1976. O/Aer anpporfr National Institutes of Ilcalth's Division of Rerearch Re- sonrtas, National Ins1NWe of Child Heallh and Human Development and the National Eye Institute. Frvm TM Jackson l.aboratory, Bar Harbor, Me. PRESENCE OF A HIOH-MOLECULAR-WFIOHT RNA AND RNA DIRI?Ctl:.l) DNA POLYMhRASE IN RABBIT IIERFI)11ARY LYMPHOSAR('OMA 10 light ol the association of type C RNA virus with lymphosareoma in severd aninul specres, it seemed pourhk that rabbits with lymlawsarcoma might harbor type C virw(es) or viral markers. To test this, the assay for t RNA-JireclcJ DNA polymerase (RDDP) was useJ as an indicator of a type (' virus in lymphosarcomalous rabbits. An K1)1)1' asurciateJ wNh partrcks Ihat harNl in the density range of type C RNA vuuses and viral cores was demon slrated in 95% of the tissues taken from the.c rabbrls. Fndu6enous KUnP a. Iivily that was sensitive to treatment with RNase was deteclcd in the crale enzyme obtained from lumorous tissues. The Kl)DP associated with the par- ticks could be distinguished from cellular DNA pulymerases by salt elutNrn /rom phosphocelluluse. The partially purifieJ enzyme pre/erred the template primers poly(rA)•(dT)r: ra and poly(rC)'(J(i)rs r, over other synthetic tem- plate primers and also utilized viral 70 S RNA as template. While this and previous studies indicate that lype C viral marken are present in rabbit tnsues, no virus from lumor celh or normal cells in culture has been is)laleJ yet Beditian, 11. (3., Foa, R. R. and Mrlrr, II. Concer Research 76:1697-469fl, 1976. Otber u.pporfr National Institutes of Health and the l1. S. Public Health Service. From The Jackson Laboratory, Bar Harbor, Me. EVIDENCF. FOR A PARTICLE-ASSOCIAT El) RNA 1)IRI'C 1 EO DNA POLYMERASI: IN RABBIT PLACENTAI. ANI) ll ft:RINF '11SSUFS Because of evidence that type C RNA viruses eaist in rabbit tis.ues, and because type C particles have been found in rabbil bla.lrc yals and utcrine cells, in primordial germ cells and in the embryonic or placental /issue o/ several other species, it seemed possibk that rabhil placental tissue, as well as pro6esia lional and es/nn uteru., could eapress type C virus(esl or viral nurkers Re ported here is the isolation of a distinct rabhit RNA-drrected DNA polymerasc (RDUP) associated with particks that band at a density characteristic of type C RNA viruses. That this enzyme is biochensically anJ bwphysically similar to the RDDP of mammalian type C RNA viruses is shown by column chromato. Rraphic eharacleristics, ten)plale primer preferences, nkrsecular weight determi- naliun, and an absolute reyuiremenl (oF the divaknt cations. 1 he methodulosy used provides the sensitivity required to delect low inlracelhrlar levels of Ihe enzyme and to dntineuish the cellular DNA polymerascs ( a. p, and y) /ronr viral RDUP. The accurrwdNed evidence imlicales that the rahhil may hartw+r an enJi>tcn.ws type C RNA vital itenome and thus conlarn viral markers Ihe presence or absence .A viral markers in Ihsues of tahhits, p,r„cul.rty slurn WII/J which has a high Incidence of lymphusarcorna, rs also hernR invcsu(ares in hopes of eluciJaling the rok of partially eapresacJ type C vrrusieal rn lumorisenesis. Bedigian, tl. O., Foa, R. R. and Mcier, H. ('nncrr Rrsr.orA 76(t2):46e7-1692, 1976. Other aupporz: National Institutes of Heallh and ll. S Public Ilealth Sctvicc From'Ihe Jackson Laboratory, Bar Ilarbor, Me.

TRANSPI A('F.N I Al. ('ARCIN(X:I:NI(' I.I I F(' I SOF DIETHYLNII RUSAMIN@ IN MI('E This suwly inve.tisates the prenatal carcinogenic c/fccls of a srn8lc Josc of diethylnitrosamine 11)FN) administered to mice at JrRercnt lunes JurinB pre8- nancy. Lung tunktrs and kukemias were observed in the o0.prin8 of DEN- trrated SWR/1 females and AKR/1 maks when the carcinogen was adminis- lered any time between the 8fteenth and iijlhteeMh Jay of Sestation. 7 he inci- dence of lung Iumors, histologically nwltiple pulmon.ry adenomas, was high- est (t77i ) when the mice were treated on day 18. Since the I)EN did not Induce any tumors in these animals when administereJ before day I S, it appears that susceptibility of the oflspring to this carcinogen begins at that time or later and that the targei organ is the lung, althou8h kukemia was common as well. Previous reporls suggest Ihat, in rals, the fetal enzyme system ncces- sary for the activation of "indirecl" alkylating carcinogens becomes operative only in later prenatal sla8es. A study of the .ynlhesis of dealkylating enzymes in fetal Iiasues at diRerenl Iimes during rs/alion could be important for es- lablishing a positive correlation between transplacenlal carcino8ersesis by these wbnances and the development in letal tissues of enzymes capable of inetabo- li:ing them. Di.aq B. A. aod A(eier. N. nk N.rr.wbun,cAa/ren 6l(10) :,s7-IBR, 1976. O1A.r aNrprtr I.eukemia Society of America, Inc. Fran The Jackson I aboratory, Bar Ilarbor, Me. THE BIOSYNTIIESIS AND BIOt.OOI('AL PROPERTIES OF 6-HYDROXYME i11YLBENZO/QlPYRENE As part of a series of studies on the properties of aryl hydrocarbon Ay- droaymethyl synthelast, this paper shows that the enzyme is present in lung and liver both as a microsomal membrane-bound form and as a soluhle form, and that each form of the enzyme is activated by a-naphthoflavone. In addilion, the blolo8ic properties of benzo(o)pyrene (BP) derivalives as carcino8enic agents in mice are studied, .nd the eRecls of cytochrome P-1S0 inhibitors on 1he side-chain methyl osidalinn and the hydro.ymethyl synlhetase rractionn are eompared. Results show that the aryl side chain methyl oaidatwrn is inhibited by the classic inhibitors of cytochrome P-450, whereas the aryl hydrusyrnethyl .ynthetase reaction functions independenlly of the cylochrome P-eS0 pathway. Also included in this paper are obser.ations on: (I) the mechanism of the synthelase reac/ion; (2) Iumori8enicity of derivatives of BP; ()) induction of the synthelase by 8P• (4) transformation of cells in eulture, and (S) Iraro- lormalion of mouse embryo cells by BP and BP in the presence of 1-henzykmi- dazok. Mexerwer, data are presented to Jemnnstrale that WI-1R human lung diploid flbroblasts IrcateJ with BP and 1-hen,ylrmidazok clone more effectively than with either ueatment alone 1he sum of these data indicates that 1-hen- zyhmKl.zule, an inhibitor of cytixhronse PIStl, in combination with 8P allows Irandormation of mou.e embryo cells al pa.sagc N anJ Ih:d thrs combrnatron al.o aRecls WI-IN cclli cluninc enicicncy Ihe rclalronship Lclwecn thFs ac tivity and Iransfurmauon is currenlly undcr .ludy in a total proRram evaluating the ruk of the mmhydroaylative pathways in carcrno8ersesn by pulycyclic aromatic hydrocarbons. Slo.rne, N. /t.. ('hen, 1/.. Diwan, B., Bedigian. R, and Mrier, It. In: FreuJenthal. R. 1. and Jones. P. W. (eds ): CurcinoRrneru. Vu1 l. I'olr- narfror .(rommtoc IlydrocurAoni: CAemi,rry, A/rraDr.lisnr, und ('a.rinozenrur, New York: Raven Pren, 1976, pp. 171-180. Other support: American Cancer Society. Frons the lkparlmenl of Biochemistry, llnivershy of lennesace ('cnter fu( Ilealth Sciences, Memphis, and The Jackson l.aboralory. Virus I eukemu Scc lion, Bar Ilarbor, Me. S11/1)11'S ON PULMONARY ARYL //YI)R(X-ARB()N IIYI)R()XYI.ASI'. ACIIVIIY IN INBRED STRAINS OF MI('E This report describes some of the mapw paramelers re8ulatrn8 the constitu- tive and )-me/hykhdanthrene (MCA)-induced kvels of pulmonary and hepatic aryl hydrocarbon hydroaylase (AIIH) activity in inbrcd slrains of mice. Inlrr tracheal instillation of 18e rg M('A in slerik 0.2% gelatin in sahne resulted in preferential induclioa of pulmonary AHH. After Irea/n.en1 with this dose rd M('A. the pulmonary AHH kvels of strains CS781./6('um, ('S7Bl./61, BAI B/ cMai, (')11/fMai and C371J1 were observed to be induced within 24 bows aller Iredmenl. Strains DBA/2Cum, AKR/l, Sll./). 1)BA/2/ and RF/I c.- pressed no such increase. At a dore of 500 pg MCA, the pulnwnary tissue of UBA/2 mice did e.press a/ourfold increase. This increase in Allll was de- termined to be quite different /eom the increase observed in ('S7B1_/6 mice by: (1) specillc activity of the enzymes; (2) genetic re`ulation; ( 1) susceptihday to inhibition by 7,8-benzoAavone, and (4) spectral propcrlies of the associated cywchromes. Thus, this paper shows that there was bolh a qualitative anJ quantitative difference between the increased levels ot A1111 ohserveJ in the AI111 "non-inducibk" mice and the increase oMerveJ in Ihe A1/11 "mJurihk" mice Moreover, in crosscs involving the ('37B/./h('rmr arwl I)BA/2('rrnr sluins of mice, pulmonary AIIH responsiveness lo inlratNacheolly administered M( A was re8u1a1cJ by a single /ulosomal dominant gene. /)n the (rasu of these rc• sulls, it seems that this genetically controlled enzyme res/wwse may play a nrajor rok in Ihe ultimate susceptibility of pulmonary tissue tu ahcmraally induced cancer. Kouri, R. Ei. rr a/. (Mkrobioloricuf A„w iarr,) Chemicaf-Diological Inre.arrlons 1): )17-711, 1976. From the Department of Biuchemiial OncoloBY, Mr.robiulogical A.ariates, BethesJa, Md. 1q '11

MACROMOI.F:('UT AR nETERMINANfS OF PLASMINO(iEN ACrIVA1OR SYN1HFSIS Neoplastic transformation of primary or early passaged 6hrohlasts is ac- companied by enhanced production of plasminogen activator ( PA ) which is probably directly related to the Iransformalion process, since increases in PA kvels do not occur after infection of fibroMuts with cytocidal viruses or kuk- emia viruses. lhe experimenta initiated bere aim to define sonic of ihe factors which may be involved in the control of plasminogen activator synthwsis. ('hick embryo fltxoblasrs infected with a temperature-sensitive mutant of Rous sar- eoma virw. Ts 6s, were selected as a model for study because sucls cells are normal by tsmrp"ic and biochemical criteria when grown a1 /1'(', but are transformed at )6'C. Since lhese phenotypic properties are /ully reversible within a few hours dler appropriate shifts in Iemperalure, this system is a very favorable one for esploring variables that might govern activalor produc- Iion. Earlier work showed that the properties of plasminogen activator are determined by the cell, rather than by the transforming agent. 7he eRects pro- duced by the addition of several metabolic inhibitors to cultures at The tirne of the shift from the permissive (16; C) to the nonpermissive (41'C) lempet- ature suggest that intracellular enzyme can be maintained at  high level i/ RNA synthesis is inhibited. 7he data also show that: (1) marked fluNualions in plasnsinofien activalor produclson occur very rapidly following temperature shifu; (2) the formation of plasminogen activator in this system is closely cor- related with the esprcaion of the vital IransforminR function; and (.I) RNA synthesis musl precede termination of activator Iormalion when cultures are shifted upward to lemperalutes which ue nonpermtutve for Iranslormalion. al- though the nature of the rekvant RNA specics rs unknown The ability of aclinomyein to limit the disappearance of mdoctble cn:ynxs following transfer to nonindueing condilions has been rcpx-rted for several syslems arwl various interpretalions have been oAered as e.planatuuns Ior the%e phenomena. '1 hese include possible decreases in the catahohsrn of specrfic proteins arnd inhibition of the synthesis of an inferred RNA species with a rapid turnover, which might function ao an inhibitor of translation. The dala presented here are as yet in- sufficient to permit either discrimination between these hypotheses or the tlimiaation of other plausible e.planalions. RI/RJw, D. R., Beal, L. P. and Rekh, E. In: Reich, E., Rifkin, D. B. and Shaw, E. (eds. ): Proteatet and elologicd Con- trof. Col/ Sprlng Ilarfior Cow/rrrnccz on Cell Prdi/craNon. Cold Spring Har- bor, N. Y.: Cold Spring tiarbor t.aboratory, 1975. vol. 2, pp. e41-1117. Other w'pr(r National Irolitules of 1lealth. From the Department of Chemical Biology, The Rockefeller Univeraity, New York. PROTP.ASF.S PRODUCED BY NORMAI. AND MALIONANT ('P.1.LS IN C(11.lURN ll+n paper jeKrilYt the crMtrol of plasminogen activators (PA) synthesis In two cell Iypes, chuk embryo /ibrobluts infected with a lemperature-stnsilive mutunt of Ruus aaucuma virus and human embryonic lung cells. Induction of I PA rcquires MHh RNA and protein symhcst. 1)cmJucuun ul PA rcquaes a ncw RNA and, perhaps, protein to be made. II the synthests o/ this dcmJucttve RNA is inhibiled, PA production can continue Irn many hours unJer aunJi Iions that are rxxntally nonpermissive. Many ddlerent eaperirncnts have in.h caled that the observed increase in production of 1'A by nenplastic cells is the resclt of Iransformation. A number of the phenutypic chan(;es asxwulcd widr transformation re dependenl, at least in parl, upon Ihe generation of plasmm by the PA secreted by these celh. One of Ihese chanRes, the lu.s o/ intracclhdar aclin containing cables in neoplastic cells, can be correlated with /he level of extracellular proleases. Moreovet, /hese same proteases can cause the dnap- pearance of actinconlaininR eabks in normal cells when supplied t.osenou.ly llterelore, plasmin accing from the ealernal sidc of the plasma membrane ap- pears to he capable oC causing the dissolution of these inlraccllular slructures 1 his paper dso attempts to demonstrate how the production of 1'A, a paurctdar component of the Obrinolylie system, may he implicated in the etiology ol diverse pathological conditioro. Ri/kin, D. Q. and Pollack, R. In: Ribbons. D. W. and Brew, K. (eds.): Pcorcolyus and PhyrioloRicd RrRu- lurion. Miami Winter SymOoaium, New York: Academic Press, Inc.. 1916, vol. 11, pp. 26)-2ti3. Other supporf: National 1s111utes of Ilealth. From the Department of Chemical Biology, The Rockeleller (/mversily. New York, and the Department of Microbiology, Slate University of New York al Stony Brook. UNUSUAL MANIFESTATIONS OF CANCFR Among the many onusual proper/ies of neoplasms, great attention has been focused recently on eclopic hormone secretion. Macromokcular /rnms u/ polypcplide hormorws, such as big AC711 and big Rastrin, have been shown to be produced by pulmonary oat eell carcrnoeus, certain Ihymumas. pan creatie islet carcinomas, and other related cells. In this investigation of ekctrrrn mieroRraphs of dysplastie ad normal human bronchial neuroerwkrcrine epuhc lial eells. two slatistically significant differences in orgaoelk tdlrastnu•lure have been uncovered which appear to be rekvant to the size of /he hormone nwlk cuks produced. Cornpared to the normal human bronchial neutoe/NharuK epithelial cells, dysplaalie epilhehal cells had smalkr (iolgi vesicles and /twet ses:reltxy Rranuks. 7hese ullrnttuctural diAcrtnces may cor.elue with nce pre/ertntial secrelion of biR ACL IL by dysldastic and neoplastrc pulnNrnary epithelium. 1t seerm, Ihert/ore. that macronNdeculat pcptide hurmures such as big A('hN or big gastrin are secreted ectuptcaay ItrHn sonic Jysplasuc aod neoplastic neuroendocrine cells partly because of underdeveloped (ialgi vcsKlcs in which the Irypsinizalion of prohormones may normally occur. Sommen, S. C. In: Finckh, t=. S. (ed ): Tbc Effctd ol Environmcnr on ('cl/r and I ruwn. Amsterdam: Eacerpta Medica. 1976, pp. 173-17N. From the Dcpanment of Pathology, l<nos Hdl Ilospital, New York 10 1 26

i COMPARISON 01: 7WO ANF_SIFiETICS FOR T11EIR EFFEC'iS ON T11E CLEARANCE RATE OF BENZO(a)PYKENE FROM MOUSE LUNGS Penlobarbital, an anesthetic used in this laboratory for the saKly of pul- monary tumor induction in mice by aromatic hydrocarbons, is a vety weak Inducer of aryl hydrocarbon hydro.ylase (Allll). Because of the possibility, however, that penlobarbital combined qith the inlratracheal instillatton of belvo(a)pyrene (B.P) could enhance the clearance rate of B,P, duplicate e.perirnents were conducted to compare the BaP ckarance rates from mouse lungs with two different types of anesthetics, pentobarbital and methosyflurane. PeMobarbital was given intraperilonedly, the mclhoayflurane by inhalation. Melhoxy8urane had a very slight eRoct on the AH11 system. There were no significant differences. however, in the clearance rates of BsP from tb: moane lunp in parallel esperiments employing these two aneslhetics. These ohserva- tions support the preferential ust of pentob.rbiUl anesthesia in these animal Nudiet on the base of its ease of administration and k.ng-Iasling eRecl, and because mortality is minimal at the proper dosage level. Fuest. A. aad Wiko.. K. (Untrrrslry of San Francisco) rrocrrdinp of the Western PAarnsarolory Society 19:32-35, 1976. From the Institute of Chemical Biology. University of San Francisco. San Fra.cisco. 11. The Rerpir.tory System FUNCTIONAL ANI) BIOCHEMICAI. FFFFCTS ON THE LUNG FOLLOWINO INHALATION OF CIOARET7E SMOKE ANI) ITS CONSTITUENTS. Ill. SERUM ANTITRYPSIN AND BRONCIIOMOTOR RESPONSES IN RATS As part of a continuing series of investigations on the bronchopulmonary effects of cigarette smoke. this report eRamines whether eaposure to cigarette amoke eta{geratn emphysemalous lesions in animals and further increases air- way resistance. Une.penedly, the data considered here indicate a possible re- latioeship between bronchomotor responses in rats and long biogenic aminea. As in sorne forms of human pulmonary emphysema, rals with evidence of et- perimenlally-induced emphysema showed decreased serum anti/rypsin activity. Additional e.ryswre ul these anrnr.ts so crgaretle snw.ke, howevcr, did not ea- agacrae Ihe lun.o.•n.l rhaners ia nce tung nor lurlher rrduce the enzyme's .clr.dt While vn..ar usha1H.••n ,auwl hr..u~lrti~.nclnclNrn in the rMrrntal a.rMr.•1. •nJ M~w~1r•.LLrw•n .u rtv enyarunul.. the nature of the , bromchumolor rc.prmse was allered by prclrcatmcnt of Ihc rrl wi/fb .yrnp. IhunurnKlic anuoe.. Ikcause nurcpinephrint arrd rAqr,munc evoke a hronchu con.uicksr rc.lr.nsc to hypotia whercas epmcphrrnc and .yncphrme claFt hn.ncho.bleluun, the authors prnttdale that preluadmg crtec/nrtrnune .lores wnh an N-rnethylaled anune (epinephrine cx syncphrinc / cau+es brrmchoJd.rtrun whilc similar preloading with nonmethylalcd amincs (rnorcpincphrinc or dopA mine) resuits in hronchoconslriction. Thn hypwhcais .cuggcsls that elnncphnne is the neurohumoral transmitter in the bronchlal muscls ul Ihe rat arrl that n can al.o he rckased by such inhalants as cigarette smoke. IIM)% niuogcn, haluthane- and aerosol propeNants. Support for Ihts new Ihcory is IKin6 oh lained hy analysis of epinephrine in the blosd draining the airways Ito, Ii., Watanabe. T., Shore, S R. and Avlado- !). ToricoloKy and Applied Pharmacology 33(3):401-a12, 1976 From the Ikparlmenl of Pharmacololly. University of Pennsylvania Sihor-4 of Medtcine. Philadelphia. 1RAC'HEOBRONCNIAL AND PULMONARY CY I OLOGIC CIIAN(;ES IN SHOCK Tracheobronchial smears from 10 palients who had developed hemorrhagic shock while undereoing general endolracheal anesthesia for suraery conlaincd mwpholosically abnormal histiocytes. These celis were overloaded with Papam- colaou slain which compressed the nuckus aeainsl the cell menrhrane. No srKh ce/Is were seen in smears from the 10 palients ased as contruls. ('ytrkhernical staining methods were undertakew to discover the comp.nilion of the suMl.ncc in the histiocytes. Tllere was no escess ingeslion of any sub.tance detecuhle by PAS. Sudan 3 or alkaline phosph.tase slarns. All cascs in .hock yieldcd smears with histiocyles which had phaeocyt'ved material whnfi was Pru.sian blue pi»ilive, indicating that the substance contained inorganic iron I he masi mum proporlion of hisliocyles containing Prusaan blue Rranulcs was 40% in slwx.k patients and only 2% in normal eonunls. flrauloRk studies cunduclcd on rals submitted to hemorrhagic shock were carrkd urd 1u invesuTalc this phenomenon. Iron-laden hisliocytes were fwrnd in the lidncys, spleen arwl IunKs of both shrkked and control animals. However, subsunu.lly more hrslur cyles containing Prussian Mue positive granules were drxovered rn the IunRs of rals in shock than in eonlrols. It may be, Iherelure. that iron is depo%rtcd in the lunp in low flow atates. Friedman-Mor, Z., CAafon, /., Katz, J. S., Gurstcrn, F., / ur ndnr/, 1/ , and Orkin, I.. R. Tbt lowrnat ol T.au,rra 16(1) : SM-62, 1976. Ofbrr ar.pp..rr: U. S. PuMic 1lealth Service. Front the Ikparlmenl of Aneslhaiology. Alhert 1'instcrn ('ulk6e of Medicine. lhe Bronx. N.Y . and the (hparlnrcnts of AneslhesK.hr6y and I'alhul.rgy, New York University Medical Center. New York. 29

0 VIRAL INCLUSION BOf)INS IN 1 RA('IiEOBRON('HtAL EPITHELIUM UF: ASYMP1OMAl1C SUBIECTS Cytoplasmic inclusion bodies have been fuunJ in the urinary tract epi- thelia of both symptomatic and asymptomatic patients, but such cclis have only been reported thus far in the respiratory tract of subjects with viral disease and bronchoeenic carcinoma. 1)uring a survey conducted for the cytodia6nosis of early bronchogenic carcinoma, howevett cytoplasmic viral inclusions were found sporadically in Iracheobronchia/ smears of aaymplomatic patients of both aeses. Is to 80 years of age, undergoing general endotracheal anesthesia for various surgical procedures. Of the ),019 smean screeneJ, 1.19G contained ciliated epithelial cells with eosirsop(silic cytoplasmic inclusion boJies. Statistical andysis did not disclose any relationship between the presence of viral inclusion bodin and other ahnormalities noled in the smears, nor was there any cor- relation between their presence and the age, sea or smoking hahits of the pa- tienu. When the frequency of occurrencs was assessed in relation to the sea- son of the year, however, 60% of all thesr smears werc found to have been collected during January through March. It is thus probable that the majority of these patients actually had asymptomalic coronavirus, respiratory syncylial virus and rhinovirus infections. According to the authon. the low postoperative complications rate noted in lhis series seems to indicate that the presence of viral inclusion bodies is not a precursor of respiratory disease, even ur,der stress corsditioos. K.ti,1. S., CAdon. /. and Turndorf, fl. C/kat 69(1):336•558, 1976. Oth.r aaspport: U. S. Public Health Service. Froae the Deparsment of Anesthesiology. New York University Medical Cenler, New York. TRACIIEOSRONCNIAL EPIiHEL1Al- MULTINUCLEATION IN PREECLAMPSIA MultinuckHed ciliated epithelial cells have been seen in the past in the sputute of patients who had ursdergone a bronchoscopy 4R hours qreviously assd. In a different sludy, in the Iracheobronchial washinp of patients under- goiwit surgery for a wide variety of estralhoracic turswrs. Now, smears were tssade from the tracheobronchial washings of 40 nonsmoking parturient women who were undergoing cesarean section or forceps delivery under general anes- thesia adminiucred via an endotracheal tube. Results showed that the smean lrorss 12 patients with preeclampsia of mwe than 24 hours' duration contained significantly nwre multimsckated ciliated cells than those of 13 healthy con- troFs, eigM women with preeclampsia of less than 24 houn' Juralicm, and seven wdnen with associated diseases other than preeclampsia. While the eaplanalion of dsis phenomenon is atill in doubl, it may be that it is related to immune mee6anisma that accompany prrectampsia. ('Adon, /. Mars. () F and Katr, I S ArcAirrr of r.rhobgy and Lasorerory Mrd.rlne IIM) 127•4211, 1976. 30 I Other aupport: U. S. Public Ilcallh Scrvicc. I rum the Department of Anesthesiology, Ncw Ymk Ilnivcr.rly MeJrcal (•emrr, New York, anJ the Ikparlmenl of AnesthesroluRy, Allxrl I in.ICm (•olksc ul Medicine. The Brona, N. Y. r.r•ANTIlRYPSIN AS A SYSIEMIC 1)1411'.RMINANF O1 I.I1NG SIRUCTURE ANI) FUNCTION The first section of this review den>,mslrares that slow, pru6resive loss ul lung parenchyma begins in mid to later life in pera.ns with a fNHnaryg.nrs Je ficiency of ar-anlilrypsin and the ZZ phentNype for this proicm, arKl Lcrhaps also in smokers with a helerotygous deficieucy of ar antiuyp.rn arsJ the M/ phenotype for this protein. lhe ar-anlitrypsin is maJc and/or JeuruyeJ in dre liver and its deficiency has been corrected by liver tranaplantatiun 1/ tbi% Jr fkiency causes the lung destruction. then ar•anlisrypsin is indeeJ a systcnuc detcrminanl of lung Wruclure and function. lividence that the tuns is a meta bolrc organ and that in this role it influences organ function el.ewhcre in dre organism is also presented here. Mos1 research on the etiology ul tMs tSenetraat ly-dctermined type of emphysema now focuxs on the hypulhean that this condition is caused by a deficiency of tr-antilryp%in, which readrs in the unre strained action of one or more enzymes on pulmonary tissucs fl(wwever, it is equally possible that ss-asNNrypsin acls primarily at a rem.NC site and sends loaic products to the lung. lndeed, il is even p.nsiMe that a, amiuypsin h.n other as yet undiscovered func/ions, or that ar•antitrypsin deficient patients have other defects that could cause emphysenu. Ilowever, this genetic disea.e remains the only solid clue to the cause rd s.wue forms of human emphysenu and may eventually enable investigalors to unJeratan'J the oriRins o1 other forms as well. Should sonse common hiochemical mechanism be proveJ, it rs pussibk that the same drugs would be effeelive in the prevention of all types ul emphysema. ' CoAen, A. s. In: Crystal, R. (ed.): The eJocAcnrkuf Rann u/ l'nbnnrro.y Fun.Hon, New York: Marcel Dekker, Inc.. 1976, pp. 717-l6(l. Other aupprf r National Neart and Lung Institute I:rum Temple Univenity Medicd School, Phd:rJclphia IIYI)ROXYPROI.IN1: ('ONl1iNIS ANI) 1'Rt11.YI. IIY/)ROXl'1 ASt'. A('IIVIIII:S IN LUNUS 01= RAIS 1-XI'11S/•1) 111 1 t1W I 1 VI 11 01: OZONE Cull.`en synlhesis in the lungs of a.luU rat" c.p.wr./ Iu l/ N ppru or-rne was studied by estimating hydrusyprufinc Illyp/ coMCns and by hdl,rwrn` nce ac/ivily of prolyl hydroRylase (P///.  cnrccd enryme rn the padhway ut aal lagen hiosynlhesis. In the early pha.es (I s_ da)s) uf u+uuc•iuJuccJ rnlury. P1/ activity was increased twrJuW over eonlr.A values und the amount ol cullasen synthesized (as estimated by Nyp formation) was double that of the nontol 31

lasennus prtqein synlhcsited. I hic rtsulleJ, by Ihc third day, in a significant increase (29% ) in lulal lung aill.rgcn Iluwev.•r, with conlmuc.l csptnorc (up 1o seven Jays), noncollagtn.ars prnlcin synlhrsis inKrcaseJ tn the p.nnt wherc 1he ralio of cullascn to noncullagen.nr. paNtin .ynlhcsrtcd was comp.trahle to that of controls. When the esp...eJ (1) R ppm Il,/ 7 Jays) •rnimals were placeJ in 611ereJ ambient air. PII activity returned 14s ntnmal in I l Jays whereas 11yp conttnt remained elevated for up to 2R days. l hcse resulls indicate that in- creased hrnR PH activity is lempwally rllated to atnrx esptnure and suR6es1 that this model may he useful in the study of lung injury and repair prucesses. Htnsain, M. Z., Cron, C. E., Mustala, M. O, and 9hatna`ar, R. S. lI/r Scicnccr I R( 9): R97-904, 1976. Other aupport: ll. S. Public Ilealth Service From the [kpattmenl o/ Internal Medreine and ('ali/urnia Prirnipte Rcsearch Ctnter, Universiry of California. Davn, and the Connective lissuc Hiochemnlry Laboratory, Universily of California Scho.rl of l)cnlntry, San I ranatco. BIOCHEMICAI. EFFF("T OF 07ONE I:XPOSURE IN RATS EXPOSEI) TO (•1(3AREfIF SMOKl: Pulmonary ghrcose 6 phosphate JchyJrugcnase I(i h PD) aclrvitics in the eylosol-rich fraction from conutri, ozone cspuseJ. ciearette<aptneJ- and aRa- rtlle-plus otune-csprncJ rats are rcported in this papcr. Whereas ntone-eapo,eJ rats show .n 11% Jecrease from the activity level (if unlrealeJ cuntrnls, rats eaposeJ Io eigaretre smule for five wecks .huw a significant 42 7 % ) increase. Subsequent esppnurc to acute. high Jnse utune /1 ppm lor 4 hr) decrcases the augmenled (i-6-PU aclivtlres to near nornial levcls, indicating that rats ta- posed lo cigarette snwke induced auRmentati.rn ol (i-h-Pl) are sull susceptiMe to (3-6-PD inhibition following ozone eapnsure. Ilowevcr, (i-6•PI) activilits in the rats eaposed to both cigarette srtwtke anJt utone remain slightly ekvated above control kvels. lhe importance o/ this enzymatic eflect wqh respect to Ihe susceptibility of smokers to oaidant eapusures is unknown. Peirce, T. H.. York, (7. K., Franli, C. E., and ('rou, C. E. Archives o/ Environmtntal Heolth 31( 61:290-292, 1976. OtArr aarpprt: U. S. Public Health Service. From the lhpartmenl o/ loternal Medicine, l/nivetsity of ('alilurnis School of Medicine. Davis. OZONE INiFRA("1/ONS WIfl1 LUNG IISSt1E. 81(X'HIiMI('Al. APPROACHES Summartting the hinchensical effects of ozone (O,) on the lung, Ihe au- thors review the molecrd.r rnechanisms of (!, inductd damaRe, the acute hieh- dose ellecu, the thromc low Jose eflccls, anJ the decreased and ra:reased sus- eepbhrlqy to lung /t, J,nute /1 is nro.t likely Ihal the moleeular mechanisms of 0, cytotocicny uperate throuRh o.idotiun and/or oaidation products, inler- actiun wilh ccllular mcmhrant% and rc.ulting Jcpu•..iun. rd rnany cclhd.u crvymalrc aclrvrtic.. Sonk e.pcruments .uggc.1 that. rn amm.J.. a wrdc varrncty td anlioaJanl .ubuances pra.vitk relative prutcttaut aRain.l //, c.pu.ute Ihat high-Jtnc eRpusures .evercly dcprc.% several crvym.rlrc :rctr.na% alnNnt ccr lainly indicates •aculc I/,.induccJ lung cytotuaiuty 1lrrwcvcr, d the annual% a1c alluweJ Io recover (rant nonlethal acute coipnsurc., .unk ol she hrnR nrctaMdr.w parameters arc greatly au`menteJ, apparently rcllccthnK she ccllular rcparatwc anJ prnldcrativc response to injury. (.uw kvel tl, caprswre ha. a siorilar cllcct wdhm ?4-2R MHrrs. According lo snnse nNUplwdoprc oh.crv.uinns. prulungcJ c.ry..rue ot cxpcrimcntal animals lo O, cau.es mrW (il.rr..is o/ luter parenchywa I he corKurrcnl increase in lysosomal entymc aclrvuy al.o cuRl;c.ts thc accunw lalrun u/ rn/lammat.ny cells, mainly macrophagt•s. in she lung Ihrs harlt.lm a: unmts /ur she reported increase in ettllaRen .ynthe.r% dutinR she catly miury and repair pluscs ('urrenl evidence alw supKcsts that a srgndirant rapccl ut sn.ceptrhrlny Nt prolonged e.ptnurt is "adaplrve" Iolrrance, a curs:entrrtarn dependent phentmrcnon whereby the mc/ahulrc auKmentatnsns inJrarJ by N..l kvel esptnures reach their maaimum two lu /otu JAys :dter the inur,d espncurr then Jrmirtish, disappearing completely rf eapu.ure is cruuinuc.I 1 hn% lar, hnw ever, thcre is only a suggestion that helpful aJaptatiuns 1o chrnmc /uw kvel 11, eRpusures occur in man. Since numerous varrables can affect an orRamsm'% resprmse to losic suMlanees, the authors recomnsenJ caution in interpretinR the overall effects of environmental pollutants starh as 1), on paramclers of IunR bioehemical aelivitin. Crort. C. E. et d. The Anrcrican lornnal o/ Mtdiciwt 60(7) :939 9)S, 1976. Othrr aupports National (astitutesol Health. Fnmt the Section of Pulnanary Medicine. Ocpartmcnt i+l Internal McJicinc. Univetsily of California Schools of Medieine, t)avis and I us Angeles . SrIMULAlION BY CIGARETTE SMOKE 1)F (iI t/1Al/llt)NI? PI:ROXIUASE SYSTEM ENZYME A('1IV11IF.S IN RA1 1 UNti (3lutathione perosidase is part of she enzyme syslcrn involved in generating reduceJ nicotinamide adenine dinuckolide phusphate 1NAI)1'I11, anJ as srMh con.litules patl of a potentially important interuehucJ ra•Juatrve anttoadant tk/cnse mechanism In erylhrascyles and hurR usatw. lhis sNrJy wve.trRatc. 1114. shnrl term, in vb•o eAects o/ espcriment.d crK,ucltc snwd.e esla,.ure on Klwa Ihiuwe peroaidase-rclaled entynse systems in prdoutnary dsarc, arwl attcmpts ao correlate any chan6es wilh histopalhobpic ohservauuns within th- ranKc ut IrRhr microseopy. Mak Spra`uc-nawky rats. 10 Jays uhl, ( aesar...n JernveJ and specific pathoEen lree, were esposed to 1) ciKarcttcs daily for 21 .rrn.ctutuvc days with a Walktn reverse-snqking eaptnurt appar,qus At the cnJ ol thar tinve, specUophotumetrie assry of IumnrRenucJ lung nssuc .ha..e.l utcrea.c.I activity of `lulathione peta.aiJase O•1Cf• 1. clutathM.oc reductasc 424% 1 am/ gIuc.ne 6 phtnphale tkhydro`cnase (1R1: / cunmparcJ lo contrra valucs 'I hrs particular kvcl of snsnke eaposnre, howevcr, diJ rwN causc Jctc.•tahk hntoloRic, lesions. It is suggested that shorl-Itrm, low level ciRatctte smuke ealwsure c.n 1i

initiate metabolic alterations in lung ceils without evoking histulodic changes detectabk by light microscopy. it is pourhle, however. that such t:ir.chemical alterations represcnt but one stage in a multistace process Ihal could eventually lead to more oven structural changes following more intense or prolonged ex- posures. Certainly. the presence of these biochemicd abnormalilies wgsest a sensitive method for assessing the effects of cigarette smoke or some of its . constituents on Ihe mammalian lung. ~ York, O. K., Peirce, T. 11., Scbwartz, L W., and Crofi, C. E. Arrhlvrz o/ Enrironnwnr.f HrdrA 31( 6):2d6-290. 1976. Other aa'prl: U. S. Public Health Service. From the Lkpartmenl of Internal Medicine, University of ('ali/ornia School of Medicine, L)avis. ASPIRIN AND EXERCISE-INDUCED ASTHMA Since exercise causes release of prostaalandin and sometimes also induces aahma, il seerm possibk that exercise-induced hronchoconslriction may be due lo the release of prostaglandin. Also, becauu acelylsalicylic acid inhibits proa- taglandin synthetase. thus diminisbrng proslaslandin rekase, it may he that n- pirin decreases or inhibits the bronchoconslriction of esercise•induced asthma. In order to lesl this hypothesis, four asymptomalic auhjecls with a history of exercise-induced asthma and a 10% or rmue drop in Forced Iapiratory Volume in one second ( Fl:V r) were selected to study the effect of aspirin on pulmonary lunction lolbwins exercise. lhe double blind method was used to administer aspirin (5100 mg) and placebo prior to exercise. Pulmonary function tests did not show any diRerence in response between aspirin or placebo. It is concluded that in Ihese four subjects aspirin did not prevent the hronchovascular response, perhaps suggesting that prostaglandins have no significant role in esercne-in- ductd asthma. The authors suggest, however. that a lar`cr series of patients muM be studied before this hypothesis can be compklely rejected. Taveira da Silva, A. M. and Ha.nosh, P. Prozt.glandinr 11(1) :71-73, 1976. From Ihe Deparunenl o( Physiology. Biophysics and Medicine, (horgelown U.iversity Schools of Medicine and Lknlislry. Washington. 1). C. L.IPOPROIFIN LIPASE IN RAT LUN(i: EFFECT OF DEXAMET/IASONE l.ipoprdein Irpase, an enzyme active in the capillary endotheliurn, regulates the uptake of blood Iriglyceride-/atty acids I ipoprotein Irpase acti.ity was de- termined in the lunes of rals slier administration of four ho.rnim:s - deaa- methasune, I thyruan, estradiol-171/ and progesterone - which hrre a known effect on hprd nxtahohsm Irpnprotem hpase aclivity, or lung sur(ac.ant synlhe- sis. In addruun, lung Irpuprolrm lipase activity was studied in diabetic and lae- tating rats Ik..mcthasone administration caused a rise of 70% in the level of , activity of lipoprulein lipase in acetone powdcr. of Iuns and a 1(1l)% uKreaK in the amount of enzyme rekascd during hcparm in/usiun into isol•rled. pcr/uscd lunp. Fnzynx aaivily was higher in lungs ut (cmales than of male rat.; how- ever. the level of activity was unaffected by estrogcn or pro6cslcrone adminiqra- lion to either mak or ovarieclomized rats. Diabetes. hypcrlhyroidism or lacla- lion did not change lipoprolein lipase activity in the (ung. 1 he conslanl preseMc of Iipoprolein lipase activity in the lung su6Ltsts that this organ is ahle tn main- tain a steady supply of Iriacylglycerol-Ia11y acids under cirarmatances that markedly impair the ability of other tissues to utilize this aubslance. Stimulation of enzyme activity by deaamelhasone could lead to increased uptake of triacyl- glyceroldatty acids by the lung and may thus be a contributing factor to coru- costeroid-induced enhanced sur(aclant synthesis. llamosh, M., Yeager, H., )r., Shechler, Y., and Hmnoz/i, P. BJocAinrico rz BIopAyrk. Act. 1)1:319-323, 1976. Other are'porf: Washinglon Hean Association. From the DeparlmeM of Physiololly ard Binphysics, and the lkpartment of Medicine, aeorgelowa Uaivenily School o( Medicine. Washinston, 1). C. INTRACYTOPLASMIC FILAMENTS IN PULMONARY I.YMPIIA/1(' I:NI)V I'HEL.IAL CELLS Earlier observations by various investigators have sue6cstcd that inlra cytop/asmic filaments may he aclinoid parts of a contractile systcm. Since fine inlracyloplasmic 8lameaN occur in pulmonary lymphatic endothelul cells. Ihe presenl investigators used tiswe biocks (rum IK neunalal rabbus to conrpare the cytochemislry and uNrastruclure of Ihese tilamen/s with myo/ilaarents of the peribronchial and pulmon.ry vascular smo.Nh muscle cells I..o types of endo. thehal filaments were observed in the pulmonary lymphatic cells: thm fllaments which lie close to the abhrminal cell membrane and thick fllaments which are dispersed throughout the cell cytoplasm. Followqrg heavy meriomyosin (HMM/ Ireatmenl, characYerislie arrowhead complexes /ornred in the thin /ymphatK endo/helial filaments as well as in the aclin filaments of the smouth musck cells. ihere was no deKetabk reaetion of 11MM with the thick elamenls. Alter incubation with EDTA. the thin Rlamenls were labilc, and the thic~ Alaments became the major filamentous component in the enduthelial cells lu smoo(h musck celh, the actin myofflaments were also labile whik Ihe larger lilamenls were stable. lhese observations supporl the hypolhesis that the acun like endu thelial lymphauc /Bamenls (orns parl of a eonlraclde systeor, whrk Ihe rhwk filaments conslitule a plastic eell•skeklon lhe significance o/ she contracule syslem in lymphatic endolhe/ial cells mighl lie in a wechamsru /or t]rc acnve regulation of the em/ulhelial interceUular junctrons and jlaps, and hence the permeability of the lymphatic endolhelial cell Irning. I.auwrrynr, ! M, Baert, 1. ard Ikl.oecker, W. Crll d Ilsrwr Rrzeorch 16)(2):11l-121, 1975. From the Katholicke Univenittit le I.euven School of Medicine. Leuven, Rc/ gium. 11

I FINE FILAMFNIS IN LYMPHA`IK' ENIX)1111i1.1AL (-Et I S Several types of cells contain fine cytoplasmic fllaments wh.ch closely resemble the myofilaments of muscle cells. 7 he staining characteristics of these various cells suggest that they may he actinoid and form part of ^ contractile system. Since fine intracytoplasmic filaments occur in lymphatic endothelial cells, the authors postulated that they belong to a contractile filamrntous sys- tem present in these cells and investig^(ed their fine structure after incubation with heavy meromyosin (HMM) and EDTA. Results of subsequent electron microscopy showed that pulmonary endolhetial cells of neonatal rabbits contain a lu6e number of fine cyloplasmic filaments measuring either 5•6 nrn (thin) or 9-10 not (thick). The thin filaments react with 11MM, forming arrowhead structures, and appear labik when treated with EDTA. 7he thick fllaments do not rtacl with 111MM and are stable in EDTA. Comparison of the thin filaments with myo8laments of smooth muscle cells suggests that the thin filaments are aclin-likc and form part of a contractile syslem, while the thick filaments con- stitute an elaswic cell skeleton. The chemical structure of these fllaments remains to be espbred It is suggested that a contractile system in lymphatic endolhelial cells might be involved in the active regulation of the endothelial inlercellular gapa or junctions and, hencc. the permeability of the lymphatic wall. Such a system may be involved aa well in other cellular ptocesses such as endocytosis, ttansport and secretion. [jrwtryns, !. U. tiaert, J. and de I.oecker, W. TAr lournaif of Cell Bidop 60( 1) I61-t67, 1976 From the I-aFwralory of IHntupathoioRy and the I alwtatory of Pathological Biochemistry. Kathuheke Umvenacet tc I euven tichixd of Medtcrne, Ituven, Belgium. ALVEOLAR TYPE 11 CEI.tS: StUOIFS ON T11E MODE OF RELEASE OF LAMELLAR BODIES Although the evidence increasingly suggests that type 11 alveolar cells are responsible for the production of pulmonary surfactant, whether they actuaUy synthesize it is still inconclusive. One of the problems remaining to be solved before this can be stated unequivocally is that of secretion. Previous sludiea suggest that lamelbr bodies are released by merocrine secretion Here, mor- phologic evidence is presented which supports the view that lamellar bodies are in fact t<leased by e^ocytoais. Freeze-fracturing and electron microscopy of rat lung tissue show that Ihere are sites at the apical surface of type 11 cells where the limiting membrane of the lamellar body is ckarly fused with the cell plasma mernbrane and that an open channel eaisb between the contents of the lamellar body and the alveolar space. At these sites, the lameltar contents eatrude into the ainpace with consequent loss of the highly compact organization of intra- cellular lamellar Msdin. The intaclnesi and continuity of the membranes can be traced for the full estent of the eaocylosis site. Freeze-etch replicas of type 11 cell membranet show depressions which may represent the siles of dis- charged lamellae Also seen are IonRue-tr-e foWs which may be cytoplasmic estrnsrons surroMrndrnil the releasing lamellar body and which may Aap over 36 the e><ncylosis pil after discharge. Fkclron phidomrcrographs of the alveolar space show disorganized Iamellar whorls which' appear to he unravelme to produce tubular myelin. In view of the unusually large size and lipid conspo srtron of larnellar bodin, ^ mechanism involving hydralion of nnrcopulys.c- charide contenls as an cid to eapulsion of lamcllar contenls is su6Rested Ryan, U. S.. Ryan, J. W. and Smith, D. S. Tirsue a Cell 70): s[17-s99. 1975. Otbe- arapportt U. S. Public Heallh Service. Harllord Foundauon and the Heart Association of Palm Beach County, Fla. From the Papanicolaou Cancer Research Instilule and the Ikpartmenl of Medi cine, University of Miami School of Medicine. Miamr, Ula. RAPID PURIF ICAT:ON OF HUMAN TRYPSIN AND CIIYMOTRYPSIN I Highly purified. matimally active enzymes are needed in order to evaluate the properties of human pancreatic proleases, particularly the stoahi.uncuK relationship between the proleinases and their inhibilors. Farlicr reports have shown that the Kunitz bovine pancreatic lrypsin inhihitor, u>,r-mcncially avail- able as Trasylol, reversibly compkaes with cationic human arypsm and chy. molrypsin I but does not inhibit any of the other human pancreas prolcinascs A one-step method for the purification of IxNh human cauonic arypsin and chymotrypsin I is described which is based on these observatiwms lhe tcchnpue involves the adsorption of the cntymes onto a Sepharose- 1 rasytol atlini!y col- umn. Subsequem elution and separation of the two enzymes is accomplished by a gradient of decreasing pH. 71ws Ireated, some samples of bovine trypiin with as little as 30% INratabk active sites have been converted to chymotrypun• free trypsin preparations with 87-90% available active sites. Johnson. 1). and Travit, I. Analytka/ Biochenrtstry 72:573-376, 1976. Ofber asrpporft National Institutes of Ilealth. From the Ikparlment of 6iochemnlry. Univcrsity of (3eorsia. Athens. IIIIMAN LEUK(K'YTE ORANUI.E kI.ASIASIi RAPII) ISt-1 A110N AND C'HARA('-II:RIZATION It has been proposed that the chronic obstructive lung disease which de veQsps in individuds deficient in it-1•proteinase mhihitor (i 1-anutrypsrn) re suits from the elaslolylic degradation of (unR alveoli by enrymcs in alveolar macrophases ^nd the granuks of polynK-rphomrckar Ieukocylcs At least lour such enzymes with different ekclrophorelic nsubdihes have been observed in the (eukocytic 6ramdes, but none has been successfully isolated. llere, the ao- t7

0 thon describe a simple twn-step procedure for isolaling gramde elaslases, based on the weal inhibitory activity of bovine Kumtz basic trypsin inhibitor ( fra- sylol) loward elanase Ihe technique involvcs allinity chruoratography o/ crude ku4ocylic granules c%tract% nn Scpharox-lrasylul, followed by ion exchange chromatography on ('M-cellulose to resolve 1he rsrrclastases All were found to he gIycopruleins I he malur furm had a carbohydrate content essenlrally com- posed of unly neutral sugars and a molecular werght near .1(),(KN1 daltons, with that of other fornis being slightly higher. Preltntinary structural analyset indicate that all the elastase isoenzymes have identical N11:-ternsinal seyuences, suff- Res/ing that Ihe observed diflerences in mobility are due to minor changes in carbohydrate content rather than lo different degrees of activation of  com- rnon zymogen Human granulocytic etastases are less active on Irgament elastin than porcine pancreatic elastase, but both are inhibited by syn,hctic elaslase active sile-ditected low molecular weight compounds, as well s by plosma a-1- prottinast inhibitor With the latter, a stable compks having a molecular weight of 7e.0()[0 dallons is formed, indicating combination in a I:1 ratio. Baugh. R. l. and Travis. /. Blochen9lstry 1S(1):6)6-d41, 1976. Other support: National Institutes of Health From the [kpartment of Biochemistry. llniversity of Georgia, Athens. HUMAN AlPF1A-I-PROIEINASE INIIIBIIOR MF('HANISM OF ACTION: EVIt)EN('E FOR A(-11VAIIUN BY LIMITED PROTEOLYSIS Although the role of human ar-proteinase inhibitor (ar-PI) in controlling tissue destruction by endogenous serine proleinases is well-known, the mech- anism by which this inactivation occurs is no( yet undetslood. To gain insight into this process, an attempt was made to clardy the steps involved during inactivation of serine proteinases by re-isolatrng the ar-PI (formerly called ar-antitrypiio) from compleses formed with porcine trypsin. llse re-isolated protein (ar-P1') was found to be noninhibilory and to have a lower molecular weight than the native inhibitor (IS,U00 vs S).000 for ar-P/). Sequence analy- sis of ar-PI• showed that an amino terminal peplide had been los1, apparently the result of cleavage at a(.ys-lhr bond. these resuHs indicate that ar-PI eaisu as a pro-inhrhiloe which is activated by limited proleolysn. As a conse- quence of this aclivation, the inhibitor traps the activating proteinase by form- ing a stable compks, presumably by acylation of the serine hydtosyl of the en- zymt to a earhosyl group o/ the inhibitor. It may be cuncluded, therefore, that limited proteolysis is the flrst slep in this inhibitory mechanism. )ohnson, b A and T.avir. l. Bu.m hi.nle d anJ H.•Phrrlt al R.icwt h l'a.n•nunr, uri..nr 72( 1) 11 19, 1976 (1/h.. ...ppo.l (nah1urr...t IIc.Id' I n.m itv 1../ N.•.Mn•...., 1 Alh[ns Iy i IS(/1 AIIUN OI: ALBUMIN FRUM WIIUI si III/MAN 1•1 ASMA ANU I RACIIUNAIIUN UF ALBUMIN-1)EI'1 1:11:1) PLASMA lhe isolalion of proleins from hhrnd pla.ma r scrurn rs ultcn .omplr.n,~d by Ihe prescnce ol albumin as a contaminam I'arolrcrdarly signdicant ro pl..~u.~ fractionation i. the lact that many prweros ut currcnt hirrh-gac mtcrc.t havc physical properties simdar to Ihose of albumrn. cspccia/ly with rcgard tu nwrkr ular weight and ekctrophorttic mobility. A previously dcs.rd.cd In.wcdua /ur the separation ol albumin from those other plasma prutcins by .J.urpta.n un tiepharosc-Blue Ikalran columns had malur dr.advantagc. In dic prc.cm paper, the au/hors recommend a modification of the affinity gcl, whrch suhsn tutcs the dye ('ihacron Blue F-)-(iA for Blue 1)auran, rc.ultUng us mrnh greater capacity for albumin (40 mg albmsin (wNrnd/nd ol gcl/ and rn a.tahlcr Sepharusc lrgarwl conjugate. Passage of whole human plasma duuuKh thn type of culumn renM.vcs about 99% of the alhumrn lhis can be yuanlnauvcly ic covered by desorption with Na.S('N. lhe allwroin-dcpkted pla.ma rs rcaddy re.olved into discrete fraclions by conventional biochemKal Iechnitsucs. In par Ircular, plasma proleins with properties similar to thuse of native hunian plasura alMmdn are readdy resolved by ionraehange cMumatagraphy u( the %ephato.e dye-Irealed plasma on DF:AE-eellulose. Even though it has been sMrwn that Ihe dye has an affinity for enzymes utilizing AMP /or reactwns auJ even tMnrgh rt also 6as been suggesled that il resembles a nucleotide in shape /hus binding 1n any protein with a"nuekotidt (old' the mechanism by which ('riracron Bluc- Sepharose interacts with human plasma albumin has not yet been determined Neverlheless, the techniqut described here is simple. ineapen.rve. easily u- pealed, and sufficiently mild Ihat denaturation is minimizcd, which makes it mosl useful for analyzing fluids with a high alhumin content. Travii. J. rt d. Biochemical Journal 117: )01-306, 1976. Ut6er sapportt NaliorullnslNules of Neallh Frons the [)epartment of Biochemislry. University of (:eurgia. Atheru and Marshfield ('linic Foundation, Marshfleld, Wn - ~ I(IN(i AN111'ROfFINASIi: A PUIIiN11Al. 1)la I NSU. A(iA1N11 I.MPIIYSI~MA 1)/:VIa.UPMFNI Eatracts of certain polymorphnnuclcar kukocytr% and .dvr.,lat macr.o phages are capable of inducing eapetirnemal cmphy.ema Ihis pri~cr tclrnts the presence of an annproteinase in canine lune lavagc whrch rs ddlcrcnt Iruur the known serum amrproteinases and might Iu/K/nul as a purcrur.l reRularur ol lung protculysis- hence aiding in Ihe dc/ense ag..io.t Ilur.c pr.~rcrna.cs dr..r may cause pulmonary emphysenu. In Ihn atuJy, IrmR antrprr.tcrnasc was e. Iracled by hronchial sahne ((19`1^.• Na(11 lav.rKe of Ire.hly r..ncd. salinr perfused dog tungs the crude anuprotema.e concenrrates prclured Ir.,m thc ./1 dug samples were tested against various protcmase. As .ould t•e .cen, dlc I

1 inhibition indes (anxwnt of inhibitor necessary to cause 50% inhihiliun of a specific entymt) of the lung antiproteina-se diflercd from the indcscs ol serum, wy bean trypsin inhibitor and ovomucoKl 1 his high molccular weight anli- prottinase was heat labile and  strong inhibitor of pancreatic elasl•rsc enteroly- sis. Although elastue ahrne or elasta.se mued with a sham protein inuillcd into a bronchus gave emphysematous ksiom, no lesions were induced when the elas/ase was Ant mised with lung anlipro(einase. Weinb.unr. G. rr a1. Anseric.n Rrriew o/ Rtrpirato.y Disease 113(2):=,3-2,5, 1976. ~ Oth.r aarpp.rlt National Heart and Lung Institute. From the Pulmonary Disease Section and Research I.aboratories, Alherl Ein- utis Medical ('enler, Philadelphia. ROLE OF LEUCOPROTEASES IN THE (iENESIS OF EMPHYSEMA Papain and other esogenous entymes with stront elaslolylic properties have been shown to produce taperrnxntal emphysema in ammah. In the search (or endogenous proleases which could damaRe alveolar walls n a way similar to papain, dogs were esptned to acronrslired homogenates of rtos poly- nwrDfwnuekar kucocytes (PMN) lhe ability of other dog cells ausd PMN from di8erenl species to produce esperrmemal emphysema in dogs was also invesligated. lhe contents of human and dog PMN, as well as dog alveolar maerophages, produced emphysema-like ksKms which were quite similar to those induced by papain, while dast monocyics and rabbit PMN did not. Fur- ther work showed that only cells which have significant proleolylic activity at neutral or alkaline p11 were capable of producing emphysema. Partial purifica- lion of the active agent was obtained from an acetone-precipitaled and desie- caKd preparaliort of the crude horrwgenate. 7 he fraction soluble in O.1 SM N.C) was most active. This impure fraction did not have eslerolytic or elasloly- tie properties when measured by the usual procedure with synthetic subslances. Tlse production of esperimental emphysema trKrk place a( the air-hrng inlertace and not by the circulatory route in this model Finally, an inhibitor for this enzyme Iraclion and pancreatic clastase has been found in lung lavage material. The inhrbitor is not alphar•antitrypsin bul has some immunologic and hiochemi- eal Nmilarities to alphas-macroooklwlin. lhe significance of these findings in relationship to human emphysema remains to be determined. Kimbel. P. and Weinb.unr, G. In: Junod, A. and [>lHaller, R(eds.): Lung Mctabo(ium, New York• Aca- demic Preas, Inc. 1975. pp. 25-41. OtAar aupp.rsr Nauo.J Heart and I.unk Institute. From the Pulnw.nary Disease Sectwrs. Albert 1-imtem Medical (-enter, Phila- dclphu 40 111. tdearl and (:irt•(.larion IN VlVO INIIIBIf1ON OF CHOLESIEROL t1PIAKE IN RAIII111 AORI'AS BY 7-K1:1(KHOI-EStERO1. Previous reports indicate that in man and sona animals. The adiLtUUn ol 7•ketocholeslerol to the perfusion fluid inhibns in rirro artcri.d chok.tcrul up lake, possibly as a result of the competitive inhibition of HM('.1'oA rcduu ta.e Still lacking, however, are published data arncernm` such inhibition in the vascular wall in riro. The esperimenb dcscrnccd here dcm.rn.rratc that u, rabbns. the intravenous injeetion of 7-kelochoksterol, rerKkrcd s,rlul.le by cooi hining it with brk ulls, inhibits aortic cholesterol uplake. 1 he effect was mN, however, as marked or as uniform as previously noted in the rn vuro espcri ments, probably because in the injecled animals plasnsa levels did nut rcach the inhibitory IhreshoM Administration by gastric mluba/iun alw failed to inhibit aurtrc clK+ksterol uplake, prob.bly for Ihe same reason. Plasrna choleslerol con cemralions plotted against plasma 7-kelochokslerol concentrations aftcr muiupk intravenous injections of Ihe latter show that there is a direct relslinnship be- tween the two s/eroids, sut"ing that they hind ds plasma lepoprolcins in  similar manner. Contrary to the previously demonstrated facl that human coro- nary arteries do not synthesize cholesterol in rirro. indicating a lack of I1M(:- (•oA reduclase aclivNy. there was significant inhibitirm of cholesterol uptake by these vessels in the presence of 7-kelochokslerol. l hus, il is rmlikely that in- hibition of this enzyme plays a significant role in Ihe inhibitiun of cholesrerol uptake by aortas or eoconary arteries. It is concluded that the effect of 7- ketocholesterol on aorsic ehokslerol uptake is mediated throu6h competitive inhibition, especially in view of the direct relationship between the plasma concentrations of the two steroids. Sarma, J. S. M., Fischer, R., Ikeda, S., and sing. R. 1. Procetdintt ol the Socitry Jor F.iperinrenraf Biology and Medicine 1 S 1: 303-306, 1976. Of6er aupportt Hoover Foundation. Norris Frwndalion, and Ross and Mcrle McCollum. From the Huntington Memorial Hospital. Pasadena, ('al., and the linivernily ul Southern California, Los Anlieks. MI'.('l1ANISM OF INIIIBIfION OF CIIOLFSIFRUI. UPIAKE BY IHE ARTI?RIAI- WALL 7 he shdies described here deal with lipid symhesrs and chulesrerol upuke in iaolatcd per/nsed coronary arteries and aorlas ot man, rabhn and PrR. and• nwrt specifically, with Ihe inhibilory effect of 7-keurchMdesacrol in fK»h rn iu..r and in viro animal preparations. /n rirro, chokslcrol uptake was siRni(knUy inhibited in human coronary arteries and aortas al pigs and rabhns by I kcto cholesterol cor.ceMratium of approaimately 7/M) nnNrks/nr1 plasma in the pcr- fusion Quid. l he inhibition was reduced when 7 kcaociwiksterol crnKCUlratnNls dropped below 50 nnsoks/m1. Lipid synthesis was never affected by this sterord In rivo, the inhibilory, eeffects of 7-ketuchuksterol in rabbits were murc ddlicull 41

! to demonstrate. lhey coukd only be shown after repealed inlravenows inicctwrns of 7-kelochoksterol soluhdiced with bile salt (Na 6lycochMAale). Ihe rssetalxslic lale of 7-ketocholeslerol and the nature of its eflect nn chulesterut are dn- cussed at length. 7 here is the possihJAy, as yet unproven, that both chulestera>) and 7-ketochoksterol actively compete for identical and specific brndins sites on the cell surface. It also might be possrb3e that an increase in 7-kctochoksterol in plasma leads to an increase in intracellular concentrations of this steroid thus inhibiting cholesterol transfer across the cellynembrane. However, definite con- clusions on the nature of inhibition must awail further experimentation. SJnt. R. l. rr al. In: Day. C. E. (ed ): Arllrrorclcrosls DrrR Di,cor.ry, New York : Plenum Publishing Corporation. 1976, pp. 4194)S. OfMr urpportr Tbe Floover and Norris Foundation. From the Nuntington Memorial Hospital, Pasadena. C'al , and the University of Southers C.IJornia, Los Angeles. LIPID METABOLISM IN PERFUSED IIUMAN AND DOG CORONARY ARTERIES Using materials per/used In ri..o. these investigators studied lipid melaho- lism In human and dog coronary arteries and in human saphemNn veins. lhe formation and uptake of Irpds in human arteries were studied under a variety of aperimental conditions, including exposure to carbon monoxide. lhe eRect of collagenase on Irpid synthesis and transport in carotid arteries ul dogs was also studied. Cholesterol uptake in human blood vessels was inhibited by the addition of 7-ketochoksterol to the basic test perfusate (human'plasma with ('ll/tboksterol and ("('I-acelate. Both nherosckrouc and normal humao coro- nary arteries incorporated /"CI-acetale into hpids but /aikd to synthesire euher cholesterol or cholesterol esten. Similar results were noted in human saplrenous veins perfused at arterial pressure. Chokslerol uptake from the perfusron fluid was dernonslraled in atherosckrotie and normal human coronary arteries as well as in human saphenous veins. Carbon mono.ide increased permr.abilily of the aterial wall to cholesterol uptake. In dog arteries e.posed to colla.qenase, marked increasn in cholesterol uptake were found, but total lipid synthesis was reduced; the relative synthesis of individual lipids remained unchanged. The tesults presented in Ihis summary paper illustrate that human coronary arleries, as well as human saphenous veins, synthesize lipids but twt cholesterol ('hot- esterol flux into the artery Is augmented by carbon monoxide and crrlla,=enase, while 7-ketochoksterol activity inhibits choksterol uptake in the arterisl wdl through competitive inhibilion. Sanna, /. S. M. rr .t. (einR, R. 1.) The American /oMroval o/ Cardfo/ojr ) 3(4):179-5 e7, 1975. O1h.r arpportt Nnrris Fouodatinn. Iloover Foundatiors and Wright Founda- twe. i From the Fluntnotton Mensnrnal Iluspital, Pasadena. Cal, and the Unive.sity of Southern California. 1 an Angeles 42 I MF ' I AI!O1 1(' FA 11: OF INGFS71:1) ANI) IN11 ( 1 sil) 7 K1i1lK'11OL1:SfERO1- IN 7HE INIA('1 RAHIII I Ilavins previ.wsly shown that 7-ketochuksecrul mhrbds cholccterol uptake into coronary arecrics in vitro and in viro- the authars now dcscnhe the late of injected and inTesled 7-keto (4-rsC) chalcslerul in the intact rabbit lhe animals were sacrificed 24 hours after admrmvratron ol the radioactive sterord, and diRerent specimens were collected fnsm the aorta, slumach, apd intestinal cunlents for lipid analysis by lhin-layer chromatography. All samplcs shuwcd measurable amuunas of radioactivity in the 7-kctocholeslertd Iractuwn as well as in the alher /rpKl fractions on the thin-layer chromalu6raphy plale Ilx aurtic uptake of 7-ketochokslerol was relatively low. '1 hcse results dcmonslrate that rahhils are capable of absorbing as well as partially metalrslrrirss 7 ket+rcholes lerul and rrnlkate that Ihey eacrete it in con/usalcd lorm, mainly through the hile. In contrast to what occurs with 7-kelucholesterol, the am.«ml of choleslead already present in the vascular wall ptobalsly determines its uptake It is suR tested, there(ore, that Iheae two ateroids may triuer drRercnt uptake rnccha nisms. Sarma,l. S. M.. Biet, R. /., Ikeda, S., and Fischer. R. Artery 2(2):ISI-a60, 1976. Ot6rr a.pport: The Noover Fouodation, Wright Foundation arsd Norris Foundation. From the Huntington Memorial Ilospital and the Iluntinelon Institutc for Ap- plied Medical Research. Pasadena. Cal., and the llnivenily of Suuthern Calr lornia School of Mediciae, Los Angeles. IN VITRO INNISITION OF CNOI-ESTERO1. UPIAKE IN Hl1MAN AND ANIMAL ARTERIES BY 7-KETOCI/O1.ISII:R()1. lhis rcport demonstrates that 7-ketochokslerol, an analog of choksterol, can inihibit the uptake of ekroks(erol by the arterial wall of scveral species Puha/ik pressure was used to perfuse human and pig coronary arterks and rabbit aortas in a rnodiQed LindberRh apparatus with blood plasma obtained from the same species. Uptake of choksler.d by f.re arterial wall was nseasured using /sl1)<hoksterol as tracer. Percent distnhutron of synthesized lipid frao tions was determined by thin-layer chromaweraphy and liquid scintillation counting. Inhibitnm of choksterol uptake by the arterial wall was studKd by Ihe inclusion of 7- ketocholeslero1/0.05 to I rmoks/nd)-in /he per/usale I tre adddi.m of 7-ketocholesterol to the perlusate reduced cholester.rl uptake by the vessel by +n average of 90'X. At eoncentralions between 0. 1 and I µnudcs/nd ol per/usate, 1 ketochoksterol inhibition appeared to be the same. Ilowever, this inhibition was reduced at concentrations of 005 rrnoks/ml. In all three species rested, inhrbr INm was present and was nu1 due to the osidatMrn u/ chaksterrrl to 7 kcw cholesterol in the perfusate. Since Ihese'resulrs suggest cuml.cthtwrn between cholesterol and 7-kelochoksterol lur binding sdcs within the vascular wall, the data also may indicate a new and diAerent approach to the prevcnliun ul cholesterol buildup in the arterial wall. 43

t I B7ng, R. !. and Sarma, 1. S. M. BiocAernicd and Biuphyricd Research Comrnunicarioru 62(1):71 I-716, 1975. OtAer srpport: Iloover Foundation and the Norris Foundation. From the Nuntinston Memorial Ilospital, Pisadena, ('al., and the University of Southern California. I.os Angeles. FACTORS AFFE(TIN(i TNE ESTERIFICATION OF LIPOPROININ CNOI.ESIEROL BY I.ECIT111N: CHOLESIEROI. ACYL TRANSFERASE Itcithin:chokslerol acyhransferase ILCAT) h the enzyme respam+ihle for the esterr/kalion of lipoprvtcin cholesterol in plasma. In order to gain more insight into the lipoprotein specificity of LCAT, studies were carried out using highly purified enzyme and pure Irpoproreins. Resutts ahowed that I.CA'T csterifkd relatively small amounts of cholesterol /rorn very 1u.r density lipo- prottim (VLD/-1. low density lipoproteins (LDl-) or high aknsily lipoproteins (IIDI.) in the presence of 5% human serum albumin (IISA)• Ilowever, when the very high density plasma /ractrun (F-4) suhstratt base was suMlNuted for the NSA. VLDI. cholesterol was esterdkd a1 rates up to tenlolJ higher than LDL or 11D1L cholesterol lhus, it appears that VI 1)1. together with some axn- pouent present in f.• may provide a highly efficienl complex resultmg in a favorable configuration of substratt Irprds lur the cruyme. David, J. S. IQ., So1o#. L. A. and l.acko, A. (7. LiJr Sciences 111(7) :701-706, 1976 OtAer support: U. S. Public Ilealth Service. From the Department of Biochemistry. Teaas College of Osteopathic Medicine. North Tesas Sale University. Iknlon, and the l.ipid Research I.abora/ory, Temple Univenity Ilealth Sciences C'enter, Philadelphia. SERUM C1101 ESTFROL ESTI:RIFICA7ION IN FAMII.IAI. HYPERBETAI.IPUPROfE1NEMIA . There is relatively little information concerning the quantitative ayKcts of lipoprotein cholesterol utilization by the enryme lecithin: choleslerol acyluana• ferase (LCATI. since previous studies have been done with puri8ed lipaprroltins and wnieated kcithin/choksterol emulsions 1he 1('AT srses intact lipo(Koleins as subslrales ln viro and, it has heen postulaled, helps maintain the normal eompotNion and ahape of plasma lipoproteirn and plasma memhrane ir.lelirity. This report deals wnfh the influence of ahm.rmal liprsprolein concenlratwnas on L('Al xlivily as r,hserved in er`h1 patients with familial hyperbtlatilx.pro- leinemia ( 1 ype I11 4rum chrdesterol estenficaliun was studied in thrx sub- jects .nd in a maitthccf control `ro4rp by rncasarin` the initial and frncliond rates ul 1('A I aruviry. Ihe lypc It paiicnts had a slightly higher mean rale ' ` I I 1 of chokaterol eslcrification (csprcaseJ as mpmulc./min/nul)v hut a siRndic.nl ly luwer fracliunal rate (capresscd as ''b/nunl th:rn the prc+unuhly,healihy cnnlrols In a sroup of nurmal chik/ren ol ( ypc II parent., howevcr both dre mean and the Irachonal rates of esterilicauam were siKnificamly lowcr than those uf healthy conlrols, indicating that the Jccrca.ed fractional rate of estenficaltirn may he a yet unrecognized early feature ol I ype II hyperbpoprolcinemra arMl that I.CAT assays may eventually aid its diagnosis in phenotypit yet unaffected kindred. Additional eaperioents using heatcJ serum as subatrate inJrcalcJ that the decreased fractional esterification rates of 1 ype /) patients were most likely due to substrate rather than eazyme deficiency. Solo#, [.. A.. Rutenherg, N. L. and l.acko, A. (:. Artery 2(l):?)e-219, 1976. OtArr srrpportr U. S. Public Ilealth Service. From Temple University Health Sciences ('enter, Philaaklphia, arxl Tesas (-ol- k6e of Osteopathic Medicine, North Tesas State Universuy, 1)cntun A METIIOD FOR TIIE PURIFICAl1ON OF MILLKiRAM QUANTff1ES OF STABLE Hl1MAN PIIOSPIIA 1II)YI ( 1101 INI:• ('HUL[:Sf1:RO1. ACYLTRANSFI:RASF. Phosphatidykholine-ehokslerd acy/trarnfera.e seems 14) pl.ry ao importarnl role in lipid metabaAism, but it has been drlhcrdl In assay in humun plasm and no previously ailempled method haa hern lound sau.l.rclory In view u/ the various problems encountered with other lechniques, however, raJNrim nrunuassay seemed pronsising. lbis report. lherelorr. akscrrhes a procedure /or processing large quantities of human plasma that yiekls mdliRram amounts ol highly parri8ed and stabk enzyme suitable for the praNluctron of antiboSi.•s -Ihis involves, successively, (NNa)_SOr fractiunaliam, citric acid treaturent, aod DEAE-etllubse and hydroayapalite chromatoKraphy, each step increasing tLt enzynre concentration so that the specifk activity ut the final preparauon is ap- proaimalely tl,(NI0 times greater than Ihal of the original plasma. '1 his pnrifird product appears to be Iree of lipoproteins, a. JeterorineJ by immunaelecttu phoresis against anlihuman serum, and is minimally cunlamiruled wnh albumin (kss than 30 r`/mg of enzyme protein) as delcnuineJ by irnnunKwhf/uamm AI 1'(', the enzyme renrains slahk for four day. Iwt most of its activity (ptl'R 1 disappears by the twentieth day. Fkclrophurcus on SS6 polyacryLmiJe Rcl yields an enzymatically active /ast muvinR hanJ, a nonactive duw muvrnR hauJ and a farnl hanJ ul albumin. Fa/racts o/ eniymarrcally active hanJs au/ (ruur 10 gels which are pooled and ta/rac/cd with 1/ /SM Na('li-ImM Na./11Y)r, 1.11 7.0, produce a single band on retkelurphureas on t1;t, polyacrylarurde gel Varma, K. (i. and Soluff. [.. A. BiaN henuh rd luu.n'd 13S : SII ).SM11, 1976 Other supp.rrt: U. S Public Ilealth 5ervice. Frrxn the l.ipid Research Laboratory, 1).partment o/ Medre,nc, Icmple tlm versity HeaUh Sciences ('enter. Philadelphia

1 CHOLF_STEROL AND /f-1 IPOPROtF.IN ON LIPID INCI.U~JONS AND LYSOSUMA1. AN[) M11(X'NONI)RIAL PERMI;Ail1I.11Y OF CULlUR1:D NEAR(' MUS('LL' ANI) FNIX)IHELIWD ('EI.I S While the nvosl univenally-accepled esplanalions of alherosrnesis involve one or more Irpid abnormalities, esperimental evidence is still scetchy, some- tirnes even contradictory. In the present etperiment, cultures of rai heart muscle ( M) and endothclioid ( E) cells were Itested with p-lipoprotein, cholesterol and (or) a cholesterol ester. The effect of cholesterol and (or) fihporrotein on the production of Irpidosis was dNermined with E-cells a/ter 24. 48. 72 and % hours. l.ipid inclusions were measured by staining with oil red O. iksth E and M cells were then evaluated for evidence of lysosomal or mitochondrial perme- sbilNy stter 72-hour treatment with 0-lipoprotein with and without adrkd cho- kalerol or cholesterol eslers. Results showed that /!-Iipoprotrin enhanced the production and maintenance of lipid inclusions. In the concentrstion employed (20mR'2r ) it alone produced no effect stler 72 hours on Iysosomal or mitochon- drial lability, but with ehoksterd, cholesterol stearale, eholeslerol oleate, or choksterol linokale (Smg%) there was a trend toward labilization. Uther in- vestiplors have reportod a relationship of cellular injury, particularly sthero- genesis, to sitered function of lysosomes or mitochondria. In view of 1his. Ihe observed chanees in endothelioid cells support the possibility of a causal re- lationship between cellular cholesterol or cholesterol ester uptake and athero- /tsesis. Wtntel. D. G.. Aeosta, D. and Kretsinger. W 9. RrsrartA Commrnitationt in ('Atm.cat /'arhufaRy and Phormacology 12(4): 789-792, 1975. Other sarpporl: U S Public Ilesdth Service and the llnrver.rty of Kansas Oenersl Research Awards. From the Department of Pharmacology and 7 oaicolop, University of Kansas School of Pharmacy, Lawtence. 711E RELAl1ONSHIP OF STRIICTURE AND FUNCTION IN HUMAN HA(3EMAN FACTOR. THE ASS(X'IATION Oh ENZYMATIC ANI) SININNQ ACTIVIiIES WIfH SI:PARAIE RE(ItONS OF T118 MOLECULE This report details the participation of the three distinct structural regions of the Hageman factor molecule in the two known biological activities of Hageman factor, namely enzymatic activity and ability to bind to negative sur- /acen. Radiolabekd Hageman (actor, functional ausys of sctivity, and immuno- logic techniques were used to probe the struclure of the native molecule. sased on the snalysis of (r.gmenu obtained by enzymatic cleavage during /luid-phaae actrvation, there are three distrncl molecular segments with different properties: (1) c regron with a molecular weight of 40INN) has binding capacity with neRa- trvely charged surlaces but no detectable enzymatic activity; (2) d region, lo- 46 I + caled between c and e(ra fiments, has a molccular weight ul 1 ZlNNI, c.n I firmly to negatively charged sur/aces, hut cuuld not hc dctectcd as a I+ esistins Ioly1KMide: (7) e region with a mulcaular werRht of 'N,/MKI h.+% zymatic activity and does not bind to ncgaltvcly char6cd surlaces. Ihe ps, ration of specific c and e antibodies is dcscrrbed as well as their use in .kh, the ekctrophurel-c characteristics of the iive (cde, cd, de, c and e) pulylw1 fragments of Hageman factor which can be isolated after selccuve prutcui cleavage. Evidence indicates that the e region, but not the c ur d, is released 1 a negatively charged sur(ace when hrwnd HsReman factor is expused lu teolytre enzymes or whole plasma, and that when this occurs in the presew normal plauna the e(ragment becomes bound to ('r esterase rnhrbitur. Corhranr, C. G. and Revsk, S. D. Thtlournd o/Cllnk.f Inrttritation 37:eS2-86(/, 1976. Other support: U. S. Public Health Service. From the Department of Eaperimental Pathok.gy, Scripps ('linic and Re.c. Foundation, La loUa, Cal. METABOLISM OF PROSTAGLANDIN F, IN 7HE PULMONARY CIRCULATION Because of ils stabk ring structute, '11-,.a-proslaglanJin 1'r.('II Pt.i was selected to test the hypothesis that enzynxs of intact rat lungs mclal- eirculating prostaglandin Fr.IP(iFr.). As de/ermincd by perlusiun lollowa' thin layer ehrotnatography, a11-P(iFr. is removed Irom the circulabun du a single passage through the pulmonary vascrdar bed pruhably by ccllul.r take. According to the dda. the volume of distribution and nxan 1r.n.a I of the radioactivity sre greater than Ihrne of an inlravauul.r marker, I destran. The hrnp retain 2S-)0% of the radwaclivity, mostly in the lotru - metabdite less polar than P(iFr. itsel(, while Ibe pulmonary venous cllh contains the same tnetabolite along with some unrnetalM.lized P(il-r. metabolite can be distinguished trom prosta`landans u( Ihe A. H. 1), FE ar+ series. It is reduced by borohydriJe, but this rcactitm does not yield Pt. Srwlium hydrotide has no eflect on the tnetaholbte, tularR uut the preserrc • (1 hydrnsy kclone gtoup in the rinestruclyde. I tre chruur.Nutral.hw I+ch.v1.+ the melabolite and its reaclinn with borohydrule are tMrac rAlKcled ot 'r hydrosy-I S-ketoprostanoie acid. Ryan, 1. W., Nierneyer, R. S. and Ryon, //. S. Prostaglandins 10(1) :101-106, 1975. Ihhs•r aupp.rtr U. S. Publrc Health ticrvice, llartlurd I ouuJarw.n and Ilean Association of Palm Reach l:owmty, Fla. From the Papanicolaou ('ancer Resrarch InstiNUe and the Ikpaurnent of M cine, University of Miami, Miami, Fla.

MEIAPOI IC A('fIVItIFS OF P1 ASMA MFMPRANF. ANI) ('AVF.OI.A1: O1 PULMONARY FNIX)1111 I.IAL ('1?1 1 S, Wll lf A NOtli ON PULMUNARY PROSIAGLANUIN SYNIIIFTASE While studying the mechanisms by which the va_snactrve polypeMides. bradykrnin, an6iotemin I and anAiotensin 11. are formed anJ then JeRradeJ, the authors came lo consider an active metabolic role (or pulmonary tndothelial ulls. A summary of the results of the esperimenls Iha1 led to this c.rnsrderatitm shows that ixadykinin and angiotensin I are metabolized during a single pas- aage through the pulnanary vascular bed. While metabolism of hradykinin yiekh biologically inactive products, metabolism of angiotensin I yields, among other products. n6iolensin I1. Angiotemin 11 is not metabolized by the lungs. Other data show that metabolism of the vasoaclive polypeplides is a.:complished by peptide hydrolase enzymes of the lungs These enzymes are ncN secreted, but appear to be attached so the lurninal surface of emkrthelral cells, especially those of capillaries and venuks. The ability of a single lung enzyme, angiolensin- converting enzyme, lo inactivale the hypolensive •gent, bradykinin. and to form Ihe hypertensive agenl, angiotcnsin I1, suggests a role of the enzyme in blood pressure homeostasis. Globular particks, approaimately the si:e of angio- lensinconversing enzyme, are associated with caveolae and undillerentialed plasma membrane of cndothelial cells. Melhods now are being developed for localizing prostaglandin synthetase at Ihe kvel of electron microscopy, because of the pharmacological dats implicating bradykinin as an aclivstcn of Ihis en- syme in the lung Ryan, l. W. and Ryan. tI. S. In: lunod, A. F. and [khalkr. R(eds ): l.nnR MeraAc,lirm. Prorrnlysis and Antlproctolyus BrorAtrnicd PAarnroaoloRy 1lanJlint of Bioacrirt SuAzt.rnrn, New York: Academic Press, 1916, pp 199-424. OtAer arpporlr U. S. Public Ileallh Service and the Ilartford Foundation. From the Pap.nicoiaou Cancer Research Institute, Miami, Fla. KININASE 11 (ANOIOTENSIN CONVERTING ENZYME) AND ENDOTNEI.IA1. C'EI.IS IN CULTURE Tl+is report represents a flne focus on the localization of kininase II using a speci/1c cell type in eulture, namely pramonary artery and •orlic emkrlhelial cells. Ekelron microscopy. immunocytochemislry, immunofltKirescent micro- scopy and kininax 11 assay were the several technique! used to further test the hypothesis that the enzyme is localized on the plasma nwrnhrane of endo- thelial cells. Results eonlfrm not only the presence of kininast 11 in endolhelial edl cultures but also its Incus on endothelial plasma mtmbranes and associated caveolat as wtll as on endorhelial projectinns lhe resrdls nl.urned with aortie ctllt IrMIK•Nt r1..1 &.n.o•v /) .t nr.r iursyut In pulmwury enJrdhehum If the I,.ng rrvtnW .. ro nu.rc l..ev.lrnl in Ihrt type uf cell, enduthchal .u. I Ryan. tl. S.. Ryan. l. W. and fhiu, A.. Advances in Eaprrirnrnral Mrdicint and BmA.Ay 7/1:217-227, 1976 Other rupp..rt: U. S. Public Ilealth ticrviic. flarlforJ t ounJawn .oJ i Ileart Assocution of Palm Beach County. 1 1a. From the Papanicolaou Cancer Research In.tilute anJ the llnrvcr.rty of Mu. School of Medicine, Miami, Fla. A SFNSIIIVF RAI)IOCNEMICAI. ASSAY FOR ANG1011:NSIN- (Y)NVI:RIIN(i ENZYME (KINASE 11) lhis study essmines Ihe reactivity of pig lung anAitNensinconverlin6 t, zyme with an anakRue of btadykinin of high specific radioaclrvily, 11'='111yr, hradykinin, in ordcr' Io develop a highly scmnive assay usin` a curnmcrtral available substrate. This anakseue is degraded by angiolensin<nnverlins tnryn to which can he separated ca.mpktely arKl recovered ncan quantitatively from iMacl substrate by calion-eschanse chromaloAraphy I n phasis was placed on sensilivNy so as /o enable Ihe identification o/ angiaeo.u converting enzyme In isolaled cell lines and cultures In order to tesl Ihr, as%.. and its actual sensilivNy, endothdial cells frum Ihc marn sleal rabl.u prAnwruo rtery were isolated on strips of cellulose acelate paper and mainlarncd rn t., lure for 118 days. The cells were washed and incuhaled in Ilanks basic sall s, lutron, p11 7.4, at 77•C for 60 mimnes with (ir==//1yr•/braJykinin (ol: pmol) in a final volume of 0.5 ml. Sampks were assayed after IS, lll saJ r minates of incubation..Hydrolysis of the s.uhslratc proceeded linearly at a ra of 0.R4 fmol/min, and the only reaction product was ir='•/llyr•At6 Incuhaus mrslurts devoid of ee11s did not degrade the suMualt Since the reactron mr tures used in Ihese taperimenls contained about I a IOr ce1H, u rnay be t•., sibk lo use this assay to measure the amount of angiotensrn converlmg tniyo, per cell. lhe assay is highly sensilive, reasunably precise. apparrnlly speall and allows the use of //r=s1(fyr•/-htadykimn at Ihe samt or luwer concentU tions of bradykinin as are Ihought to occur in viro Ilowever, while it is th possible to measure the small quantities o/ an6wnensin-convcrlinR enzyme ., eountered in small•seak cultures of pulmunary tn.lothclul cells, certain mtrtli ealKms of the present assay may be requirtJ to ehminate thc mfluente of odu peptidases /or testing biological flnids or lissue homuccnatcs (-hiu, A. T., Ryan, 1. W., Ryan. U. S., and IAner, 1: F. Bi.x-htnrical lournd 149:297-)00, 1973. O(bror suPl.nrf: U S. Public 1leallh Service. /t:olforJ I ounJ.diun aud rl Veterans Adminislra•ion. Fuan Ihe Papanicula.Nr ('ancer Reseerth Imlqwc, anJ thc Ikparlmco/ . MeJicine, Unrversily of Miami. Mi.nu. 1lunda. an.l the Ilypcrlcn.Nrn Rtscar. (horatory, Velerans AJminnuauun lloynlal •nJ the 1)cparuncnl of Nn themislry, ('ase Western Reserve Umversny, ( levelarul 49 49

I i FUR7tl1:R EVII)I;N('k ON lllIi SI1N('F1.1111.AR SIIFS (11: KININASE 11 IAN(;IOII:NSIN ('UNVIiR1/N(i h.NZYMF) Indirect evidence suR6ests that circutalins hradykinin and ansiutensin I are metabolized by enzymcs located alon` the lumrnal surface ol the prdmunary endothelial cells A recently rsulated enzyme Irom pig lung (:mRiutrnsin-cnn- verting enzyme or kminase II I has the sante bradykinin and anRKrtcn.in I me- labolizin` capxity as the rntact lung and,shows the ssme sekclivity. Antibodies to pure preparatrons of the pig lung enzyme have been preparcd and used to localize the celhdar and subcellular siks of the enzyme in intacl luu`s and in pulnwnary endrNhchal cells in culture. Subsequent studies having shown that antibody bound to A-microperoaidase (it-MP, a heme-octapeplide of cyto- chrorne c) is more suitable for ekctron microscopy, the authrxs report on the use of t-MP conjugated to antikimnase 11 using bis-succmyl succinate. '/he im- munocytochemical results provide further support for the hypothesis Ihat cir- culating bradykimn and angiolensin I are metabolized by enzynsea on the luminal surface of pulmonary endothclul cells. Other data indicate that aortic endothelial cells also have kininye 11 on their surfaces. 7 hat this may he true of endotlsclial cells from other vascular beds as well would not lessen the im- portanoe of the pulmonary vascular bed as the primary site for inacrivation of circulating bradykinin and for anRiorensin 11 formalion, as the lungs :rrt unique in that their venous effluent enters the systemic anerial circulation. Ryan. l. W., Ryan, U. S., Schultz. D. R.. Day, A. R., and Oorer, F. E. Adrancrz in Etptrimrnml Mrdicmc and Biolney 70:213-21), 1976. Other aarpport: U S. Public Heatth Service. llartford Foundation. Veterans Administratron, and the Heart Association of Palm County. Fla. From the Papanicotaou Cancer Research Institute and University of Miami School of Medicine, Miami. Florrda, and the Cleveland Veterans H(,%pital and Department of Biochemistry. Case Western Reserve University. Ckveland. FORMATION OF ANGIOTENSIN III BY ANOIOTENSIN- CONVERTINO ENZYME In this biochemical study. rates of hydrolysis of either angiotensin I or des-Aspr-anNio/ensin I by pig lung angiMensintonvertinN enzyme were mea- wred in terms of the formation of the dipeplide His(histidine) l.eu(leucine) by an automated Ouo.escence technique. F.cepl for studies for estimating Kw and V.,, each substrate waa used in a flnal concentration of 23 or 30 rM and re- actions were begun by adding antjiotemintonvcrling enzyme at a flnal correen- Iration of 0.2e6 ng of prwein/ml 11 has been known that anotioa!nsin 11 is fornsed by metabolic conversion of anNiotensin 1, and this paper shows that angiotensin 111, a prnsrMe primary secrctoloollue of aWoslerone, is formed from des-Asp'-angio/tm/n I by .ngiolrmineonverling enryme. l/se K„ (1/ pM) of this reaction rs one third of that taw the converswrn of angiolcmin I into angio- tennn 11 /n view of thr drAerent pharmKOklgrcal effects of angrotensin 1) and 111, whether converting enzyme has a greater affimty for des-Asp'-anaiotensin I than fur angiowen.on I could -well he a questiun of physioloeical sisniflcance. 50 I SMruld dcs-Asp'-angiotensin I and ansiolcnsin I uccur in eyuimotar yu.ntni. Ihe formation of an aldosterone secrNosocue would be favored over the /. malion of a pressor agent. A higher dlinity of the enzyme for des Atip' .ns. lensin I is favored by data showin6 that ansiotensin I and 11. bradykinut a. peptide BI'Pa, are better inhibitors of the hydrulysis of an6rotcnsin I th.n the hydrolysis of des-Aspr-angiotensin I. Chiu, A. T. er af. (Ryan. U. S.) Biochemical Journal 133:1119-192, 1976. Other support: U. S. Public Flcalth Service and the Veterans Adminmuf au.,, Frorn the Papanicolaou Cancer Research Inuitulc and 1)cpartmeM o/ McJaiu Universily of Miami. Miami. Florida, Department of Hirkhemis/ry. Ifnivrrsi of ('okxado Medical Center. lknver, and Veterans Administralion Ifuspu Cleveland. EFFECTS OF PLATELETS ANf) CER*t A1N PI.A f E1 1: f(-(1MPONI N~ ON (IROWTH OF CULTURED HUMAN ENl)UIIII:LIAI. ('[I.IS Iluman and nonhuman endolhelial cells have recently been grown succe, fully in tissue culture, opening the way for a study of factors that may inMren their replication rale. One report indicales that the presence of platelets in tt nutrient medium enhances endothelial cell growth. This has been con6rrne here and lhese observations have been ealended to include the dlecb of cells platelet components on endothelial replication in the heliel that plalekl% u :ertain of their components might influence this growth rate in rivo as wcll !n vitro. Various agents were tested fur their possible eflects on endolhelral c- growlh. Results show that isolated platelets and two platelet components. AI - and serolonin, each enhance such growth in tissue culture, while epinephn- has no discernible effect. Isolated platelets had the greatest eflect, and AI had a eorscenlratioa•deperdenl one. At equimolar concerdratiom, serulonin w less stimulating than ADP and its effect did not seem to be praxluced thrwq direct enhancement of eeN metabolism. Platekls are known to partocipate Ihe formation of Ihrombi and 'wrdircetly damage tndothelium. bur they ml well play a rok in Ihe healing of vascular lesions by enhancing the growth • endolhelial cells that will cover the wound area Knowledge of such a IwKu0 nuy permit reinterpretation of the inlluence ul infused plalekts and pl.ul components un the henwstalie potential a/ IhromMrcyrupemc patients and ao mah. Ihus, this newly•discovered growth enhancing etlect .r1 platelcls on end thelial cells may have far-reaching implicalwrm rn oor undcrstandin` ul rl mechanisms of heMoulasis. Ihromboas and atherosclerosis. Saba. S R. and Mason. R. G. T'brom6osb Rrrrarrh 7:s07-N12, 1975. Otikrr support: National Institute for Arthritis. Mctalwdbsm and Ih`c.u' t)iseases. From the [kpartment of Pdhulogy. UniversAy of North ('arufina. ('hapcl 11611 411

70 .~..1k .J.. INTFRA('17ON (11`INE ('HFiMOR1iFI.1:X AND Ttlli Plll MONARY INFLAr1ON RF.FIEX IN 111E REG(11.A"IION OF ('ORONARY ('IRCULAiION IN CONSCIOUS DOGS The goal of this slrNly was to esamine the relative roks of pulmonary in- flation and chcnsorcfka control of the coronary vascular hed in conscious dogs by comparing 11) the responses to inlracsroliJ adnrinislralinn of nicotine when the dogs could increase ventilation and,when ventilation wxi controlled, and (2) the eRects of forced mechanical hyperinflalion, hyperinAatian induced by chemoreflea stimulation and. Mully, apoManeous hyperinflati..n. When the heart rate was conslanl. Inlracarolidly administered nicotine induced an in- creaae in the depth of respiralion followed closely by an rncrease in Iale dias- lolic coronary fiow and a reduction in Ia1t diastolic coronary resistance. After belrrcceploe and chdinergic hlockade, a similar coronary dilation in response to nicotine occurred only when ventdstion was albwed Io increase. Ilowever, when venlilation was conlydkd, intracaroliJly administered nicotine increased coronary resistance after combined bels-receplor and cholinergic blockades. T7re refka coronary dilation was not observed after carotid sinus nerve section or dter alphsrreceploe blockade. Thus, the results of this study propose a new aspeel of re/k coronary control in conscious animals. Nicolioe stimulation of the carotid chemorelka results in a Nriking coronary dilation that has two components. lhe minor component involves a chemoreflex with its efferent pathway in the vagi. The major eomponent of coronary dilation (ollows an in- erease in 1he depth of respiralion, and its effcrent component appears to involve withdrawal of alpha adrenergrc conurictor tone. An almost identical period of refka coronary dilation followed either forced mechanical or spontaneous hyperinflalion in the conscious dog. Yanrr, S. F. nd McRitchie, R. 1. Cirrrfarion RerrartA 77:661•67), 1975. Other a.'porfs U. S. Public Ilcaith Service and she American Medical As- socialion. Frorn the Depsrlmenl of Medicine. /larvard Medical School and IYIer Bent Brigham Nospaal, she Ikp.rtmenl of Cardiology. Chihlren's Hospitsl Medical Center, Bopon, and the New England Regional Primate Research Cenler, Southborough, Mass. CNRONIC SMOKING IN AN ANIMAI. MUDI:1.: EFFI:('IS ON CI.OTf 1N(l AND hIBRINOLYSIS lhe effects of chronic smokins on the csr,liovasculsr system were studied prospectively in beagles maintained on a chronic smut<inR program /nr 1• monlhs ('Io/1ing and flMinolytic tests were carrred out he/nre the initiation of sn.ukrng and were repeated at 6. 12 and IN monlhs in Ihe unancathclited lesl animah and their curre,ponJrng conlr4d. At the end ol Ihe IN month smoking pernMl, flhrrnnRrn wrmwer utcs snd platth•t /unatiun were JelermineJ An aJ- Jilronal Rtouip of /our dogs un high IrpKl diet and chronic smoking program S2 I lor nine months was included in the lihrrmigcn Jc.•ry ,ludres only Re,ut INrweJ Iha1 al the end of 111 ownlhs the .nwkcrs apfxarcJ to h.rve srsnrfi,a~~ ly .tkrrler clrNlin6 Iime than the control 6ruup. I lie ,arnc pattcrn ol argnifir.rn. at IM rnonlh, between the two groups ,vas lound lor partial thannlxrpl.r,r 1rme, with a borderline difference al su month,. Also at the awl o( she sna, ins period, the turnover rate of fihrinugen was found srsnificanmly increased the chronic smoker, particularly when combined with high lipid diet It wuu aeenm, lherclore, that the data obtained Irom this small gruup nl mmd, su gest si`nilicant enhancement of the coagrJalion mechani,m rn the snwku group. Moschos, C. B.. Ahmed, S. S., Lahiri, K., and RcKan, 1'. 1. Arhr..n.lc.o.ir 21:1)7-112, 1976. Other n.pperlt lhe Ameriun Medical Assucialion I:ducation and Re,can Foundation. From the Department of Medicine, ('olk6e of Medicine anJ 1)entistry New )ersey, New )ersey Medical School. Newark. ('ARI)IOVASCULAR EFFECTS OF I.UNG-II;RM ('1(iARff1E SMOKING AND NICOl7NE ADMINISIRA11/1N The relative rarity of angina peetoris among smokers with cardiac Ji.ca• su66ests that myocardial aNeralions may contribute so climcal aluwamahu, independently of coronary vascular disease. Beagks. selecteJ lur Iheir characte- istic relative lack o( coronary atherosekrosis -- which could obscure a drte, myocardial response - were studied Io determine il k/t venuiclt lunctirn, uu, phobsic features ted composition were allered by proton6cd crRarette smrrkrni (Nxervatiuns on young beaRks which inhakd cigarette snNrke over long perwvl of time (up to 22 months) were compared with data Irom another group rf ceiving an equivalent daily amourN of inlramu.cularly adnumsaered nicotine r determine i( the cardiac responses of she first JuRs were at Ita,l partially a Iribulabk to the alkaloid. All Ihe animals were slso matched ataiml a conu, group. Heart rate, stroke vdume, left venuicular end dr.stolrc pres,ure +n volume, as well as inlrsvenlricrdar eonJrKlrun unre. did ma drQer ar6nilicanrl in the three groups. l.el1 venlticular eieclNrn hxliun was hrRhe,l in the rui trols, lower in the dogs Ihst snwrleJ anJ k,west amunt thr.se Rrven nKVrun, r'c,phc simdar values /or enJ draslolie v.nal.ks in the lhrec Rruups the hr Jerivative u/ k/l venuicular pressure (JP/Jt I n,anrducd la,r Mr anJ .dlcrlur followcd Ihe sanse pattern. Although mcan sorlic pressrut was sr6nd'waurl elevated in fwwh taperimeutal eroups, thcre wss no arRndkaru cunclauoo +rd Ihe eunlraclility indices RrJuetion of a(/erlnad Irs nnrnral level. JrJ nat allr. thc abnormal venlricular pcrlurmance. Ilyprruophy, rn0ammation arMl «lluld uluaslnktural abnwmditics were shsent, arnJ myuc.rJral lip«I anJ caur+n uuu po.rtwrn were n.umsl. Since intentilial /ihro,rt was evwkm rn all espenment. animals, h.rwcver, an alleralion of e1aNrc clemems may lic of.crarive lhcv cardiovascular ahnormalilies appear 1o he predrmuruntly JepcoJenl un the nK.) trne content o1 cigaretlei. S1

! Ahmed, S. S.. Moxhos, C. B., Lyons, M. M., Oldewurtel, II. A., ('oumbis, R. 1., and Rctan. T. l. The Amerkan lournol of CarJioloRy J7(1) :))-40, 1976. Other arrpport: American Medical Association Education and Research Foundation. From the Departments of Medicine and Pallwlo`y. College of Medicine and DentiWy of New Jersey. New leney Medical School. Newark. IV. Nearropharrrt.colo6y and Physiology BEIIAVIORAI. EFFECTS AND BINDING AFFINITIES OF TWO STEREOISOMERIC PSYCHOTOMIMETIC (iLYCOLATES As part of a long-range study simed at investigating the structure-activity rrlationships of the psycholomirnNrc glycolate esters, interest has focused on the stereoconflkurational aspects of the drugs and the physicochernical nature of the pharmacophore. It has been shown that the availability of the non- bonded electron pair on the heterocyclic N is a critical factor in detcrmining psyehotomimetic potency, and that the conformation of the heterocyclic rin& and other sleric faclors can affect accessibility of the eleclron pair to a pharma- eophore. Because of the recent observatiun that Aeirro- and levo-isomers have differing central nervous system polencies. the authors attempted here to de- termine whether the diflerenct belween the 1wo enantiomers could be uaed to characterize the pharmacophore. A special cumpnter-controlled pro6rarn was used to compare the behavioral effects in cats of d- and l-)-quinuclidinyl benzil- ate (QB). Psychophysical parameters such as the fate of response and tendency to use Ihe left or right paw for kver presaing were used primarily to evaluate drug efficacy. The drug did no1 alter auditory threshold or the percentage of erron in kver pressing. The f-isomer was at least 100 times more potent than the J form, both having a similar qualitative effect. After a single dose of /-QB (3/.R/kg), the cats' performance did not return to normal until 3-7 days later. Pretreatment of the animals with atropine (2mg/kg), an .ntimuscarinic agent, did not prevent the behavioral effects of t-QB. The binding affinity of the Iwo isorners for synaptic tnembranes and phosphatidyl serine was identical. 1( is suggested that the interaction of I.QB with the active memtxsne site ma•/ induce a eonHgurational change that Is either not reversibk or requires metabolic turn- over of some protein or other trrokcule comprising the phsrmscophore. Lowy. K.. Abood. L. G. and Rsines. H. lournd of Nerrorrlcncc Rtrearch 2:157-165. 1976 Other supportr U S Public Iteallh Scrvice. From the ('enter for Hrain Research and Ikpartment of Biochemistry. Univer- sity of Rochester McAKal ('emcr, Rochester, N. Y. 54 a I:NIiAN('1=.MI:.Nr OF S"IIiRl'.OSPI:(-111(' UI'M11? HINUIN(7 10 NI•.URAI. MI:MHRANES BY PIIOSPIfAIII)YI. ti1:RIN1; Previouc studies based on evidence indicating that a prolcolipid tractwrn from brain t.asue may he the opiale pharrnaa.plurre havc shuwn that phos phalidyl serine. a malur acidic lipid of rnammalian biomemhrane, hinds o«u phine. Ihis binding is skreospecific only and appcars to he relalcd to Ihe phrr macotosic potency of the opiate. though it is recognizably different /rom Ihe high affinity binding of opiates to brain usare demonstrated by uther inve.u 6aturs. Since it seemed possible that phosphatidyl scrrne. in the lornr of a cuot plea with some membrarwus prolein, miEht hc a component ol thc uputc pharmacophorc, the authors tesled its esogenwrs effect nd that of other lopedt on the high affinity stereospecific binding ol 'Il drhydruniurphine to variour memMan.rus preparslions of rat brain. Ihe addition of phosphatodyl .cnne to suspensinns of either synaptic membranes or a mecruaumal fraction siRndicandy enhanced both high and lower affinity binding. 1'hrrphatidyl elharNdamene nnd Iya+phosphalidyl ethandamine inhibited opiate bindrn`. Sullaudes enhamcd n sli`h11y Mst other acidic lipids and lecithin had no effect Incubation of '11 damyl phusphatidyl serine with the microaomal fraction eaahlished that enouKh eso6enous lipid associstes with the neural membrane Iu accrwnt fur the cn hancement of opiate binding. The results, which are dixusscd Irom the stand point of the modi(icalion of membrane lipids anml their pusible significance lur the binding of opiates and other lipnds, suggest thal phosphatid)I senne may be an important component of the opiate pharmacophore. 1 hat the hieh at linity opiate binding is primarily associated with the synaptic memhrane is sull inconclusive, however, in spite of the facl that the mK1oxM1al IraclrM cun lained synaptic components. AAooe. L. G. and Takeda. F. tnropean lowrna/ o/ rAarmacofoRy 39:71-77, 1976. Other aupport: U. S. Public Health Service. From the Center for Brain Research and the Ikpanmenl of Hirxhemistry, Uni- venity of Rochester Medical Center, Ruchesrer, N. Y. PRFPARAI7(1N AND CHARACTFRI7.AIION UF ('A1('It1M- BINIIIN(i AND OTHER HYDROP11UH1(' PRO'l1 INS I RUM SYNAPII(: MEMBRANES Several types of eakium-binding protein% have been isulalcd lrum varaurs animal ti»ues in an effort to elucidate the trde of C'a' ' w ce/Iular fun.doo 7 hese authors have focused Ihtir interest on the characternstics and tK.ad.lc lunction of hydrophobic proteins derived Itom isrdated synapuc mrrnbta.K•s uf mammalian brains. In the present sludy. preparative acrylamide `cl ekclrophu resis in dodccyl sultate was used to acparatc and purily a numt.er of com ponents of the hydrophobic eomplca. (he irsdrvNhul hands wcrc 11Kn analyrr.l for N terminal amino aeids, amino acid cumpusilaro and Lehn.k pruflk, as wrll as for their ability to bind ('a' ' and Al P. I he maprr ellurt. Iwrwevet, was de voted to the putificdion and characterization of the ('a' '-AinJin6 prulein which It .

0 has a mokcular weight of about I h(KX), a binding capacily of 4 Ca' '/mulecule, and a Km of 1.5 X 14) ". but represents only 6% ot the total protein in the lraclion studied. It is an acidic tryptic peptide derived from this patticular protcin which seems responsible for its Ca' '-hinding activity. luJRin6 lrssm Ihc pcpti.k map- pinR. there appears to he a similar acidic componcnt in a number of proteins et<- hibitinR Ca ••-bindinR. Binding of Al P was asvrcialeJ mainly with the high molecular weight proteins, particularly Ihose which consisted of rtumeruus basic tryptic peplides. ` Abood. L. G. er at. diocAlmktt et eiopl4yska Acta 44):414-427, 1976. Other aupporft U. S. Public Health Service. From the Center for Brain Research and the Department of Biochemiury. Uni- versity of Rochester Medical ('enter, Rochester, N. Y. EFFFd'T OF CHRONIC ADMINIS1RAl1UN OF NI('OIINE ON MeTASOLIC RESPONSES AND INIFS(INAL GLUC'OSE 1RANSPORI Nicotine induces several adaptive changes in the physiologic system, in- chrding alteraliom in the turnover rate of catechtslamincs. Since norepinephrine and other amines are implrcaleJ in the central reRnlatron of hMKly lemperalure, part of this study anemp/s to .ktermine the mllurnce of prulunsrJ niculhne ad- minisUNion upon the raf's IhcrmorrRulaany rcafrm.ea tievera/ other phyaio- bgic paramelers werc eeamineJ a wrll Animals ,nl.•.teJ with fi,,e suhcutancrws nicotine doses (1 mR/kRl daily /ar acveial wecks dnplayeJ a numhcr of adap- live reactions, including incrcased ress.t.n.c Iu hyporhermia Altcr luur weeks of IreNmenl, while osy6en conwmpuun remaineJ masrmal, cooling unrc in- creased significantly as did the blood preswre. llowever, the continuous decline in heart rale (which al the end of Ave weeks was unquestionably lower than in the saline-treated conarols) may have been a compensatory respon.e to the ele- vated systolic blood pressure. Initially reduced, food intake waa notably greater in the sialh week without any effect on the growth ra1e, suggesting a possible enhanoement of the gluco,e absorptive ability of the inlesline. Actually, ikal Rlucose absorplion rose significantly hu1 independently of any changes in 6lucose melabolism by intestinal mucosal tissue. lhat this may indicane increased glu- oosa transport is supported by the fact that a nonnxtaboti:ed ghKwe analogue was also absorbed mwe rapidly following chronic nicotine Ireatment. 1 his is cowsWeM with the oMendwn that, after an rnnial decline, oaygen cnnsump- lion did not differ from control values. E.cept for the adrends, which were heavier, there were no significant changes in organ weights or morphology. SAapN. Q., van Seaumonl. W. and Elkrt, M. S. Drug Addlcrlon 4.127-1)6, 1971. Otber.uppor(r U S Public Heaph Service. Fnwn the Department of Phyvolusy. Saint I ouis llniversity School of MeJi- cine, St I 114,1111 56 I lil:l'1'.('T (1F NICOIINE ON SERUM SUCRI.IIN ANI) I X(N RINI'. 1'ANCRI?AlIC SE('R1:1 IUN Nicotine appears /o markedly dcpre.% c.octinc pancrcatic resp,mscs to, esotcnous secrelin anJ, it has been .uRgestcJ, a/.u may inhrbu the rckasc ul sccietrn in response to intestinal acidification 1lowevcr, nnc claaic atudy Jeal ing with smoking and the alimentary tract IaikJ 1o reveal any elfecl of nicotme on pancreatic .ccrelions. Because of Ihn Jr.crcpancy anJ the clinical imirir- tatKe of a possible ibhibi/ory effect of mcoline, the.e authurs have eaamincJ the inlhrcncc of nicoline alkaloid on H('1-atimulalcd secrelrn relcaae anJ on secrettn induced pancreatic secrelions. In Jogs with pancreahc fiupdas. inua durnlenal in/usion of IICI (9.6 mEa/)0 min) induced the rekase of immonu reactive secretin (IRS). Pancreatic flow ra1e, as well as brcattrmate and prwem secretiom, were stimulated either by intestinal acidification or rn/uvon of eso genous secretin (1.01U/kgfhr/. Nicoline (/oOpR/kg/hrl inluscJ together wuh 11CI delayed Ihe appearance of peak IRS c.nsccnttationa by ahuul 2(/ minulcs Nicotine had no eflecl, however, on the total amrntnt of IRti rclca.cd nor «as the observed delay ac=ompanied by a similar delay in the appeatrnce of peak bicarbonate output. Funhermore, nicotine diJ not affect either the pancreatic secretory function stimulated by NCI and esosenou% secrctm or the metaMdic clearance of the latter. Since nast of the dogs studieJ initially ahrrweJ an increase in pancreatic secretions rather than a decrease, feslhrwinR a pattern produced by parasympathetic stimulation, il may be that the elfecla of nicoline observed here are largely due to a similar mech.nism. In view of the inc.msislcnt resulls obtained in various laboratories because of the slightly ditferent esperimenlal prohr.Ja useJ, it is the aulhon' opinion that far-reaching corsclusiom as to the effect ut nicotine on duodenal uktr formation are not warranted at this time. Soden. G. e/ at. The American Journal oJ Digestive Dbraser 21(11):974-977, 1976. Other aupportt U. S. Public Health Service and the Natiooal InslNules .if Health. From the Department of Medicine and the General C'Imical Research ('enter. Temple llniversily Health Sciences Center, Philadelphia, and the Ikpartment of Surgery. College of Medicine and Ikmistry of New )etsey, Ru1Ren MeJnal School, Piscataway. /)It1RNA1. 1)11:F1'R/?N('I:S IN AL11!RI•) BRAIN t IIYI)R(/XYIRYI'1 AMINI:-RI:I.A11:1) RkC31ONAl. PROIUIN SYN1111:SIX lhe rate of cerebral protein synlhc.is in the rrMknl brain varies with the regional and suhcellular contenls of endogemnn t hyJroitytryhramine 0 1111, This study esamines the relNwmhip between diurnal varutwms in cndosemHn S-H 1' levels and microsomal protein synlhe•ra in the male 1/;ati nuanc hraur. and astempls to determine Mrw ehangc•s in this interacrNrn can he mAuentcJ at two opposite points in the diurnal cycle (M A M and II 1' M). Muuoauoul pnNein aynlheail which was evidenl in the corleR and reticrns of the limhK area in the evening when endogenous 3-111 levels were low, diminished rn dse 57

maxnins when the 5-11 T levels were high. F/cctroconvul.ive .hock, which ek- vales S-N 1', inJuccJ srgni6cantly higher amnunls, especially in 1he furehrain, when applied in the evening lhis response was acconlpanrcJ by a curresponJ- ing decrease in protein synthcsis 1'orcbraln 5-11 T increments resultinR /rons the administralion of h-methyl-tctrahydro-ficartnrlirx were regionally specilic for different phases of the diurnal cyclc, hut the negative correlation between micros<xnal pnNein synthesis and tissue 3-111 still prevailcJ. ReJaceJ S•IIT Conlents following Irealment with p<Morophenysalamne were associated with increased microsunul protein synthesis in the morninR, which wa.-. consistent with the greater magnitude of 3-NT depletion at this time. Ihe accunn11a1eJ dala supporl the hypothesis that microsonul protein synlhesis in various re`iorn of the mouse brain depends upon S-FIT kvels. lhis relationship, mnremer, is dependent on diurnal variatiuns in endogenous 3-111 contenl, or on Jiurnal Jif- lerences in the magnitude of 3•Nl' alterations effected through their physiologic or pharmacoloRic variabks. The aulhor aho suggest% that sonx of tlse pro(eins, whose synthesis is diurnally, as well as S•I1T, dcpendenl, could well comprise en:ymes with a significanl role in the process within the central nervous system and in the regulation of the metabolism of amines and other tnolecuks of si6- nifkance for brain function. Einnan. W. e. lorrnar Ae PAaernacoloPlr (Paris) 6( 7): l I 1-122, 1975. From Queem ('olkge of the ('ity l)nrvenily of New York, Flushing. PROTF.CTION 8Y NI('OTINF I ROM RFHAVIORAI. f)ISRlIPl1ON CAUSED BY RI-.11('ULAR FORMAIION CIIMUT ATION IN TIIE RAT Chronic nicotine lrealmenl has been shown to improve performance of a visual allemion Ia.k by ra1s. Short trains of appropriate levels of electrical eur- rent delivered to the rcRIKUIar formation (RF) disrupt ongoing conditioned be- havior, presumahly because The induced slale is one of general overarousal. If the RF and limbic systems are nsulually inhibitory, then Ihe hyperatimulaled state resulting from increased RF activation might be counteracted by increased limbic aelivalion. This study repreaenls a preliminary investiRaliam of nico/ine'a e/lkacy (through its supposed effects on limbic slruclurp) as an antagonist of the behavioral dnruptinn resulling from RF slimulalion. Male Spralerle•1)awky talt with ekctraKks permanently implanted in their mesencephalic reticular for- malion, were trained lo perform a viswl attention task. Short trains of electric eurrenl delivered to the RF effectively blocked performance in a revcrsihk anJ reproducibk fashion. Suhcutaneous adminiuration of nicotine (I()O pgrkg, the base dissolved in saline) served to attenuate the hehavioral Jisruplion causeal by reticular stimulation. The wltRestion that it is a nicotine•inJaceJ limbic system activation si.hich anlaRolires the behavioral disruption eauseJ by eleclrically- induced reticular overactivalem is JiscuvseJ l alensinn of Iheec resrdls to hu- man smoking hehavinr, fur(hrrnmre, auRRcds a physinloRie mechanism to ae- tAwrnl (ur nw..r.nf rll .dntnW.ualu.o, uarurly Iha1 the snwkcr uray I.e atlcmpN- rnst 1.. manqrd.fr 6.- ul.r..r ara,u.al v1a1r 1h.rv, a N ampeiv.hlc Iha1 une u.rb.l~.nj nr.......y..n an...1n.il Lrh.....r .. I/.r IfV/e lu rHhNa' an RI' aCtiva. r«.n le.rl a. ......d..vJ .n a h.l..u.n...,.r,.1 ..r •naw.m alale inappr..lnute fnr elfective behav'lor, and to engender what might he concidcrcJ a ttate nt u.eln behavioral arousal. Nelsen, 1. M.. Pelley. K. and Goldsrein, L. Pban.racoloiry eiochernls(ry-i eeharior 1.749-754, 1975. From the Department of Psychiatry. College ad Medicine and Ihnlislry r, New Jersey. Rutgers Medical School. Plscalaway. NI(Y)TINE-LIKE A(TIONS OF cis•MEfANI('OI'IN1: ANI) rrans-Mfi I ANICOTINE Isolated rabbit aortic strips and ileal se`menls were employed in a aon pa/ativc aludy of the biological actrvities ol air- arwl r.nnc melanitohne Aaurd ing Io the dala, both isomen have a nicotme•like e/1ec1 on these preparation I his interpretation is supported in part by Ihe facl that heaamclhonnrm, ciK auw phenololamine, reserpine, anJ atropine blocked or reduceJ Ihe contracuk r, sponse produced by the melanicoline iw.nlcra in a manner similar to thal prt viously observed in nicoline studies. 1)ose retpr.n.c studie% on the arntir .Iro showed that reans-metanicoline is significantly Icaa active than nrcu(ine, w1u1 cit-melanicoline is the kasl active of all. When four pyriJuws comppNmda ( pyridylacetie acid. N-(I-pyridylacelyl)Flycine, nicolinuric acid and rrunr 4(' pyridyl)-3-butenoic acid) were also tested for baNh aRoni.l and antagamla at livily, lhey had no slimldalory effect on either phyaiologic preparalion Pr. Irealmenl with either pyridylacelic acid or N-( l pyridylacetyl161ycine redrrcc the contractile response of aorlic atrips to rracn melankaline nKdentely I markeJly, while pretreatmenl with rrans•4-(1-pyrrJyl l• l hwerwric acid drJ s only slightly. Thus, the present data ckarly indicate that /he metahulnur r nicotine ard metanicoline 10 l-pyridylaceuc acid and i1s Rlycine conjugate accompanied by great loss of adrenergic and chadincrltrc activity. Wihion, K. L., Ir., Chang. R. S. L., Bowman. t:. R.. and Mr A.nnh, 11.. Jr. The lowrnal o/ Pbarnsarology and Eaperirnenrnl IherapeuNcr 19611) 681 A'/l 1976. Other adpport: American Tobacco ('ompany, American Medical AsuKral«), Education Research Foundation and National Inati(rues of Ilcallh. From The Deparlmen/ of Pharmacology. MeJrcal ('ollcee of Vueinia, Richmum PROM)S141) ROLE OF 7FIli Pt A('/'NI nl l 11(/1 /N1 Rti1(' %YS11 r•1 IN 'I IIE R1:(iU1./1'l1ON OF 1•fil'A1. I:ROW l lt ANIr IH VI IO1•MI N I All Ihree componenls of a eholinerSk sysla•rn, acclyl. hnhne (M'h) . h. line acelyllran+ferase (ChA)acelylch.4inevlcra.e IA('hl-*), ure prescnl in it human placenta. Ihey are hrcalireJ in The syncy(n.uaryrlw.hl..u an.l lhcu aaunu, vary with ecslaliamd devekrpmcnl. A('h Icvclt arc Inpho.n. Lemg luw at caul Rcstatiun periods (6 to /1 weeks) and at Ihc Imre ol paraururrn. v.hrk calnb" ing a peak at miJ pregnancy (16 to 211 wccksl ('hA and A('hl: acUvurea au, SM 59

relate well with A( h Icvels. In addition. Ihcra arc JiRcrences in the pLaccnlal uplake of suhstances which are drpcrxlcnl on the scstatiunal al;c. 1)iphcnyl- hydanloin uplake is high during early and Lrte scsutiun and low JurrnK the midsesration period Ihe same patlern holds lur the uptake ul 2 amrnraao- bulyric acid. 1loth of these compounds are conccnlralcd by the placcnlal Usaue, and their uptakcs fluctuate inversely to the development of Ihc placental cho- linergic system. lhe Restaiiun-dependent variation of the placental chodrnertic syslem, as well as the placental uptake and trans/er of various suMtances. inJi- cate a possible rok of the cholinerlic system in the modulation and mainle- nance of placental function and. subsequently, fetal growth and Jcvekrpnsent. Harbison, R. D, Oiubadewo, 1.. Dwivedi. C., and Smtrr, Q. V. R. In: Morselli• P. 1. ,(iaratlini, S. and Sereni, F(eds. ): Hwi.- and Therapruric ArPrcn o/ Perinaral PAacnsaroluty, New York: Raven Prea, 197S, pp. 107- 111. OtAer.rrprt: U. S. Public Health Service. From the Department of Pharmacology and ('enter in Tosicolo6Y. Department of Siochemistry, VanderbiN University School of Medicine, Nashvilk, Tenn. i HUMAN PI.ACF.NfAI. CHOI INERGIC SYSFF.M (1CCl1RRFN('F, DISTRI6UTION AND VARIATI(1N WI1/1 (:FF.SrA11ONA1- A(iE OF ACETYI.CHOLINti IN HUMAN PI.A('F.N1A Various puhlnhed observations rndreale that all three comptrneuls of the cholinerific system, acetykholrne (A('h)-Irke activily, cholrne acetyltuans/erase (ChA) and acelykholrneslerase (A( hE) are present in the human placenta. While human placental ('h^ .nd A('hl: have been characterrred• 'Ihe com- ponents of the ACh like activity in the human placenta have not In this study, gas chromatography was used to ascertain that the major component of the ACh•like activity of the term placenta was ACh (112 nm/g of wet lissue). This result was confirmed by the separation of ACh from other quarleinary am- mmmiwn compounds by column chromatography. Although the placenta could be stored at 4'C for a number of days without significant loss of A('h• freezing and thawing of the placenta destroyed ACh. indicating that ACh is hound with- in rnemhranes. While therte were high concentrations of ACh in all segments of the plaunta. the concentric segments nerst to the umbilical cord and the pe- ripheral segment had lower concentralions of ACh than the central concentric segsnents. the very segments where villi are localized. ACh content varied with gesta/ional age• the placental ACh kvels being higher during the second tri- mesler of preNnan.y than during the first and third trimeslers. 1 hese observa- lions indicale that ACh is possdsly I«alired in the syncyliotrophoblast. Since ChA had a similar pattern of variation with gestational ate, it would xem that the placental cholinergie system is fully formed at the early fetal period of his- tinNenesis and functwnal muwation. llte sum of these observations indicates that the placental cholinersic system may play a significant role in the regula- tton ol the translrxt of wutrk..u ar.d chemicals across the ayncytntrophobla.t and thereby rergulNea the fetal growth Sarrry. S V R cr d . ,. I Bu.rlumicaf Pharnrorology 2S(4):425-411, 1976. Ilth.r support: U. S. Public Health Service. From the Ikpartmenls of Pharmacology and Bimhcmntry, Vanderhilt (/oi versity School of Medicine. Nashvilk, lenn. NORMA1. FUNCf1ONS OF TIIE THYRO11) (;LANI) (tF l llt PYUMY (IOAT 7he purpose of this study was to gather hitherto unavadabk irdnunatw,n about the normal physiologic and biochemical values for thyroid /uncuon in the pygmy [oat which is ealensively used as a laboratray animal model Nnrm.l vahres were obtained for blood serum PBI, I s inJe., 7 s, and ch.rkacrul in a colony of 31 pygmy goats of miaed sea and a`e. Serum PHI values avcrated Q 11 1 2 pg/ IOU ml with no significant sea ditlerences. 7 he mean 1 s rndcs and Tr values were 1.1 f 0.) and 7.2 t 1.1 rg/ I(X) m) respectively. No sea Jrfkr ences were noted in these values. The toean serum choksterol value was 9011 m`/ 100 ml, and sea diQerenus were apparent. In /emaks, the average level was significantly lower (tN.1 tssg/ 100 ml; n=44/ than in intact maks (97.4 mg/100 ml; n=t) and it was 11iN lower in castrated males (69.2 ma/ 100 ml; n=6). Serum cholesterol values, however, consistently and aisni/kamly increased with age in the femaks, an uneaplained phenomemm also oMerved in humaro. The animals studied showed no evidence of either thyroid mal/unc lion or pituitary thyrotropic deficiency. Castro, A. rt d. Laboratory Anirnd Sckwrt 23(3) :727-)70, 1973. Other a..r'ortr Florida luveni)e Diabetic Research Foundation. From the Department of Medicine. University of Miami School of Medicine. Miami, Fla., and the Heart Research Laboratory, Universily of Oregon Medical School, Portland. V. /ntw.vnolo)jy and Aduptine Mt•chaniam. 1(1('At_IZAI ION OF ANOIO7tiNSIN ('ONVI?R I/N(i L'NlYME (KININASI: 11). I. PRI:PARAl1ON OF ANIIH()I)Y•111•MF'.- O(TAPEP7IDE CONIUUATES Antibodies to pute preparalions of pig luns anRrutcmrn cunverunR enryroe (kininase 11) have been prepared and ne now being used to Lelp hwalue 1hc eellular and subcellular ailes of kininase 11 in inlact IunR anJ in luns cells in culture. I his paper describes a melhrrl of conjugating these anuMrhes to a peroaidase marker suitable (or election mrcrouopy In the work presented here• a heme-octapcplide (d-microperoaidase. M-MP/ derived Irons cyto.hrome a' 61 60

I I was coupled to antibody in a two-step procedure using a hifursctinnal active esler, his succinyl succinate In the ftrsl step. 8-MP, which has only une reao tive amine, was reacted with an eacess of his-auccinyl succinate to yield A-MP- succinyl succinate, a stable compound which can he stored. In a second step, the remaining active ester was used for coupling to reactive amines of the anti- body. The conjugate consists of 1-6-2-J MP moieties per nlilxxly. Using Ihese procedurn, Ihe formation of compka polymers is avoided Fach molecule of conjupte posxsses both immunoreadivity and pern4K/a1K activity. The con- jugate has been used to kxalize anitiolensin-converting enzyme (kininase 11) along the plasma membrane and associaled caveolae of pig aortic endolhelial oella in culture. Ryan, l. W.. Day, A. R.. Schultz, D. R., Ryan, U. S.. Chung, A.. Marlborough, D. 1, and L]wer, F. E. Tbzre d C-ell tll l):111-121, 1976. Other aupport: U. S. Public Health Service. Nartford Foundation. Veterans Administration and the Heart Association of Palm Beach County. Fla. From the Papanicolnou Cancer Research Institute and the Departmcnl of Medi- cine. University of Miami School of Medicine, Miami. Fbrida; Hypertension Research Laboratory. Veterans Administration Nospilal and the Lkpartment of Biochemistry. Case Western Reserve University. Cleveland. LOCAL.IZATION OF ANGIOTENSIN CONVERTING ENZYME (KININASE 11). 11. IMMUN(X:YI(X'HEMISTRY AND IMMUNOFLUORESCENCE In this part of the localization study of angiolensin<onverting enzyme (kininase 11), the cellular and subcellular sites of the enzyme in lung tissue and endothelial cells in culture were esamincd by immunocytochemical and im- munofluorescence techniques. lAe immunocylochemicai studies at the electron microscope level used goat anti.(pig lung angiotensin-convcrting enzyme) coupled to 11-MP (11-microperoaidne) via `lutaraldehyde or to Il-MP (II- microperotidase) via a bifunctional active ester, bis-.uccinyl succinate. The Ialter conjugate, which does not contain comples polymen, has been charac- Ierized in detail in terms of immunoreaclivity and perosidati: activity. Both conjugates yield similar rewlls: antiiMensin converting enzyme appears to be localized along the luminal surface of pulmonary endothelial cells. I:ndothelial cells of large and small vessels react with the antibody conju`ah•s, but reactions are most prominent at the level of the capillaries and venuks. The conjugates ate also reactive with pig lung and aortic endothelial cells in nwnolayer culture. These cell lines were shown to possess converting enzyme which is capable of inactivating hradykinin and of converting antiolensin I to its pMent homolog. anitiotensin 11 ('onverting enzyme was also shown 14) be associated with pul- monary enrkrlhclul cells in culture by direct and indirect immunofluoreseenee. 'Ihese results support the conclusion that circulating bradykimn can he inacli- vsed and .ngrotensm can he activated by n enzyme siluated on the luminal 62 i surface of prJmonary erKlolhelial cells. I he ability of pulawnary convcr,m/ zyme to inactivate a hypotensive suhslarscc. br•rdykinin, and to form a h~ lensive substance, anitiMensin 11, may he.pcak a role of thc enzyme in 1,pressure homeostasis. Ryan. U. S. rr ol. Tissur A Crll d(1) :123-113, 1976 Oth" aupport: U. S. Public Iieallh Service. Nartford FrNrndation nd Heart Association of Palm Beach County, Ila. Frnm the Papanicolaou Cancer Research Instilule and the Ihpartmcnt of ht cine, University of Miami School of Medreine, Miami, Fla A('('FS.SORY Si'LEENS IN DOMESTIC RABBI fS (()RYCTOLAGUS cYnlculYJ). II. INCREASED FREQUEN('Y IN HFMAiOt(X;ICAI. L)ISEASFS AND EXPERIMENTAL. INDU('lION WI111 PNENYLHYDRA2INE This report documeals Ihe association of increased frequency of acce" spleens with ( I) certain inherited hematological diseaxs in rabbi/s and (2/ production of accessory spleens by phenylhydrazine. In this sludy, dala (ron, bred rabbit strains of comnan ancestry with a high frequency of heredn autoimmune hernolylic anemia or lymphosarcorna showed lhal all of rh strains also had a high frequency of accessory spkens. A/olal of 21% ' rahbita in these slrains, though phenotypically mxmal, had g/obulin coi (Coombs'-posAivc) erylhrocytes. These findings supported otwervalions of creased frequency of accessory spleens in human beings with similar dnc, and suggested that accessory splccns, rather than being passive developne, arwmalies, represent physiological responses lo demand for phallncy/ic capa, provided by the reliculoendolhelial system in the spleen. 7 hrs hypothesis I, dicted that simula/ioe of the basic defect might yickl a lahoralnry model ' the induction and study of accessory spleens in rabhils. 1'henylhydrazine • used to alter hematopoiesis and 'tl was shown that the Irequency ol access, spleens in 80 rabbits so treated was 13'R, as compared to a naturally occun- frequency of about 7.1%. The rabbit phenylhydrazine mrde/ oflen the tt direct proof that a biological basis esis/s for the indrrclNSn of accessory splcn in response to hemato/o6ical Hrns. No evidence was luruwl to rclate anev p.r re or cardiovascular uwmalies to the increased frequency of access- spleens in the rabbits. Weisbroth. S. N. rf a/. (Mrirr, N.) Trrarololry I l ( ) ) :23 )-262, 1976. Or6rr aupportr National Institutes of Ilealrh From Ihe Division of Labora/ory Animal Resources, Healrh ti.rences Cenr, Stale University of New York, Stony Brouk, and '1he Jackson I atwrralory. N Narbor, Me. Rt

a, in Vt. Epidemiology THE RELAl1ONSIa1P OF SCIENTIFIC OBIE(-TIVES TO POPULATION SELECTION AND AT1R111ON IN LON(:ITUDINAL STUDIES: TIIE CASE OF TNE NORMAfIVE AGING SlUUY The Normative Aging Study (NAS), a lifelong study of ?.2s0 initially healthy mak suhjecu, bas been active for 12 years now, and the issue of at- trition has become import.nl because of the nature of the original population. Since the NAS subjecb were selected on the basis of scientiflc objectives, rat represenlaliveneas, it seerrsed tseceasary to assesa attrition rates and to determine whether attritiors had changed the populalion i characteristics. Over Its firat dosen years, results showed the. NAS had a remarkahk capacity to retain ils population with a IS annual a11ri1ion lor all causes and a 0 5% annual a11ri- lion due to loss of interest. No remarkabk hedth, age or social ddferenlials were found between those who remained in the NAS and the small group that kft, ard it seemed likely that the initial highly selective nature of the popula- tioq as dictated by acientiflc objectives, reduced both attrition and any differ- ence between the smaN number of dropouts and those who remained in the study. Tbus. the original population characlerrstlcs have been maintained since the inception of the Study in 196) and it appears that the problem of attrition in longitudinal studies should be viewed in the broader conleat of scientifk objecli.n. Roae. C. t.., amrf. R. aed Srreuer, W. T. The CcroluoJotirr 16(6) :3o1)-S 16, 1976 OtAer wpprtt Medical Research Service of the Veterans Administration. From the Normative Aging Sludy, Veterans Administration Outpatient Clinic. Boston, aed Nelknic CdkNe, Brookline. Mass. RELATIONS OF AGE AND PERSONALf1Y DIMENSIONS TO COGNITIVE ABILITY FACTORS This study eaamined the relation between three cognitive ability (aclors - Information Processing Ability (IPA), Manual Detterity (MD) and Pattern Analysis Capability (PAC) - and three penonalily dimensions - An.iely, Ealraversion and Openess to Fspcricnce -- in 969 mak volunteers ranging in age from 25 to 112. Also, since most measures of these cognitive ability factors have shown cross-secliocsal dcclincs in perlormance with increasing aee, this study tried to determine whether Ihlse dechnes might be mediated by person- alily factors. On the whole, results showed small but significant re'ations be- tween penonalrty and cognitive ability facton, mcs.t of which could not he ac- counted for wkly by educatron or social class Anaious subjects scraed lower than adjusted sul.lects in all three lorms of intellrtence, whrk sul.lecrs more open to eaperrence .cured higher than chned minded subjects on IPA and PAC. Introverts +fw+wed some superNnrty uver estraverls. Mrl only in PAC. lust as in nther sturlres. younRer suhlccts Renerally did better than older oncs, hut aev- rral hylxithcvrcd interactrnns of age and personality were nut found lhis su/- I gealt that while penonalily and inlellisencc are suhtly rel.rtr•J. Ihc dccline in ar1snilive lunclionins with age cannot hc altributcd to the inlhrcnce of pcr- sonality variables. (-otta, P. T., Jr. cl d. Jnurnal rr/ Gernnrolocy ) I(6) :667-669, 1976. Other aupp~.rtr Medicd Research Service of the Veterans Adminnlrahon and the Natwnai Instilules of lleakh. From the Normative Aging Study. Veterans Administratron Outpancm (linrc, Boston. SMOKIN(I ('I SSATION AN/) SI:X ROLE (-UNVERGIN( I: As noted in previous reports, smoking rales are hecomm6 mnre simdx among males and females. TMs sludy nuw esamines whether hlurrmN or dc crcase in set role diQerentials as a result uf social change al.u aRects mak and femak smoking cessalion rates. As shown by the literaurre, there already is convergence in proportion of srrKtkers anN.nt males and fenules Irum older w younger age cohorls. suggesting an equalilarian shift over tuuc Ihal a srnular effect would he evident in giving-up snwskins seenred a likely pnsihility. I ur- Ihermore, convergence in responsiveness Irs intcrpersonal mllucnces due lu the equalilarian shift forms the basis for another hypMhesis acroa aee. Iroru older to younser, there is a diminished scs differential when NnPk1ng ce%tatNM is related to interpersonal (ac/ors. Age cohort analyses on a nstNm:d prohahdny sample of current and former smokers ( N=).)61) stronKly supprnted these hypothe•.es. In addition, Ihe data presented here may have significant iorphca. lions for research on aging. Rouf. R. and Rore, C. L. Journal o/ Ncalrh and Socld acAavJor 17(1) : 3)-61, 1976. Other supporrr NNronaf Ckaringhouse fur Smokine and llcalth and the Veterans Administration Normative Aging Study. From the Veterans Administration Outpatient Clinic, Brnlon, and llcllenic Co/, lete, Brookline. Mass. 1% 111F. IN("RI:ASED RISK OF MY(N'Akl)IAI INI Ak(~turN IN (l(:ARI`.1-11i SMOKERS UUE '1O PSY('11111 (><il('A1 I Rn111' AN AI'IFMPIFI) I:XPI(IRA11ON USINI: /'SY('sl(11u1i11'AI t)111:Sf1ONNA1Rl: RFtiPUNSES Ihis paper e.plores the question of whether the relatmnship lK•n.ecn rrFa relte smoking and myocardial in/araion miRhe he ha.cd on underlyrnN psy.hu logical ddlerences rather than on snwrkiae itaelf hr thrs cpidcroruluRrcal samdy, cigarette snwking assessed in a mu11rp1Aasic health chcckup was almust twiac as (tequenl in 222 men who snbsequent]y develul.ed a firat myrM.rdral m/aratHm Ihan it was in 2?11 controh Also given at this health (hcckul. werc Iti psy.h.r logical quesliunnaire items that had been derrved mosdy /rum the Mumesnla

! Mulliphasic Pcrsonrhty Inventory. 1ur nKxt /')JiI", nr 610) o( ihe items- the case conttol Jdlcrenccs wcre parallt•I tu the smokcr•mrosmokcr d-Ocrcnccs. Subdividing the study group acaurJmR to their resp+nscs Io such rtcros farkd to eliminate the apparent rtlatn.n of snKrkinP Io suhscynent infarclios in rmtst subjects. Howevcr. as many as one-lrarrth of ncc suhjacas cvNdJ he iJcntifted by the qttestionnaire as subjects in whom smokmR was not predictive oi in/arclion. The present data raise the possibility that the irnportance of smoking :n a coro- nary rnsk (aelor varres with psychological status. Although crgarene smokin` remained a risk (actor in most subjccls, the investiRators did identify a snbsroup in whom it apparently was not a predictor of myocardial inlarction. Frltdman, G. D. er at. rttrtnrirt 1Neditlnt 4(4)=326•S)2, 1975. O/Arr.rpportr National Institutes of Health and the Kaixr Foundation Re- search Institute. From the Departmcnt of Medical Methods Research and Department of Medi- cine. Kaiser•Permanente Medical Care Program. Oakland. Cal Active Projects Following is a list of the principal investigators, or institutions, of projects under way or activated in the period since the prcviuus Reprrrt, together with the respective project titles. (-ompleted projccts are listed in a later section. PRINCIPAL IN V PSi1CATOR OR aNSilTiRION LEO 0. AeoOD. PIs.D.. Troleaor of elocAcrwirrcy and frdn RtstrcA. Cen- ter for Srain Researct,. The University of Rochester Medical Center, Roches• Icr, N. Y. JOSEPH C. ARCOS D.Sc. Professor of Ar.dicrne. Tulane Uniatnity School of Medicine, New Orkaw. DOMINOO M. AVIADO. MD. Pro/ts- sr of tA.nn.cdop UniversAr of Pennsylvania School U Medicine, Pfil- adclphia. CARL O. sECKER, M.D. Associ.rt rrolessar of rr/bfop. CorneR Uni- .ersitr Medieal College. Nesr York. WIt.L1AM E. tlENEDICT, M.D.. Assbr- anr Professor of Yedi.rrits. Universiry of Soutbern Cdrfornit School of Mtdi- eiee. Division of HenWolo6y and Med- kal Ocnetics, Childrerts Hospital of Los Angeles. Lo. Angeke. RICIIARD 1. sINO. M.D. Professor of A(tdklnt. University of SoMlserw Cali- fornia School of Medicine, Lo. An- ~eIts; Visiting Associate M 1Nonctdor.l l'nrinttrln[, Califorwia Institute of Technology; Director of Cardiology and Intramural Medicine. HwMington Memorial Itospta1. Pasadena. Cal. (I1/1:NTHFR SI)UPN, M D, Arwc6crt I.ulcsuw of Mtdirlnt; Assin.wt Drrrc- nw, (aner.l (Tink.l R.st.rtA Center. lemple University IleaNh SekncesCen- Ier, PAiladelphia. RAYMOND SO!SSEt. Ph.D.. Associ.rt Dic.croc, Normative Aging Srnd), Vet- ecans Adminiuratfon OWp.ueM Clink. eoaton. A. SONIA sU/ST, M D.. A»isr.nt lro• sor of Mtdiclnt and IAysioloey. ~lnniversity of ()reaon Ileah6 Sciences Center. Portland. PROJF.CT T1TLE ('Aolrneraic correlales of hehavirw Synergistic effects of pu/ycyclic hydrocar- bons and niuosamrncs in pulmonary earcino~ enew. Potential repressors of metat+olie aclivation of nitrosamincs Influence of cigarette smose on putmu nary empsysema and bronchospnm Inresliaalion o/ 1ht rule of alltrly to tohsceo eonsliruenas in the parMraene+n of arteriosckrosis and myocardial in farction Malignant transformaion, mu/agennn and Ahrinolysin produclion of er{arrne smoke cnndcnsate fractions Inhibition of ehalesurol upule by sr Itries in rirru and in rrr.. Ff/ctt uf nKutint and tipcctNr smolr on sttrrtln ltcrction A smoking research proiect in the Nou mative Aging Study lhe rtde of alpha I anritrypein deficiency as a rnk factor in Ihe developnent of chronic airways uh.unctrun

PRINCIPAL INV[SflGATOR PROJECT T1TLL PRIN(:IFAI. IN V ESi1GATOR PROJE(:T iiiLE OR INSIiTl/i1ON OR INSTITUTION ALBERTCASTRO, PH D., Dirrcrw, Ilur- Nicotine in blood: detcction by radio- WILI IAM H. FISHMAN Pw D Pro- Cance h t f l hi h ntont Ls.rarocy; Asu.ciarr f.o/esior of Medicine University of Miami immunoassay , . ., /tssor of %rAolop•; Direrrr, Tufts r p e w eno ype pro i may prc. c sage broncholenic cancer . School of Medicine. Miami. FL. Cenr er RncarrA Center. Tuf1s Uni• versity School of Medicine, aoston. JACK CHAt.ON, M D., Asiociatt rro/es• ar of AnenAeniolop, Nc. York Uni- .ersNy Medical CeMer, New York. CHARLES O. COCIIRANP MD.. A(rn.- 1er, Scrt'~s Clinic and.Researc<t Foua- dNior~ L /olla, Cd. Fpidernidosy of tracheobrortchial mulli- ` .uckalion The mediation of inRammalory injury of liawe LARS FRIBFRG, M.D.. rro/rssor and C'Anirman. Dtprrncen/ o/ Enrfron- mcntal Nygitnr, 7Ue Kvduuka Inui- lute, Stockholm. EprdcmtologKal sludKs on the new Swrd ish twin registry Causes of death in relation lo smokinl hahits and u/her bchavuxal and en vironmemal factors. A study on the Swedish lwin Re/islry ALLEN B. CO/1F.N, M D, Pw D., Aa• a.ortue rro/cuw of ilfelk/ne, Temple Uaivenil Ileah• Scietsce. Ceacr, PWdcl~ia. PAUL T. COSTA, Jr. PsM D.. Aau.ciate Professor oJ I'+ycholoty. Utr.ertrty of 1/anKfssselts N tloron. CARROLL t' CROSS. M.D., Atsoc/are rro/eaaow of Medicine onI Nun.aw rAysiofop; Dilrecrr. Sccrion o/ rul- nsorsry Jlfedirint. Uniserwty o( Cali• focsia School of Medicine, Davis. DAVID W. CRUMPACKER, Psr D, J'rn- /n.or and CA.irwraw. Dcp.rrwanr of EnvMowwwnfal. Poplarlow and Org.- w/0rrk t/ofopy, Us.ersity of Coiorndo. Doolder. H. FRED DOWNEY Pw D.. Auiaanr lroJeuor of PAys/oJodl. University of Tefa. Health Science Center ar Dallas; Dfrecrr, Crdbr.scul.r RrarrcA, Car- dioFuMsow.ry Institwe, Methodist Hc. F11.1 oI D.R.c, Dallas. NrALTER 111. PSSMAN, M D. Pn.D., Pro/rsaoe of riycholoty owI /iochtm- brry, QIreens Cdkp of tAe City Uni- .es.ily of New York. Flarine. HUO1/ E. EVANS, M D. Director. De- prrnwn( of rtd/alrica, Jewish (loyi• UJ .nd Medical ('eMer of Brooklyn, ltrooklyn, N. V. HANS 1. EYSFNCK, Psr D., D.Sc tro- /esaor of rrrcAolory. InNilute oi Psy• chiatry. Um+ersity of Lawdon, l.on- dnw OAl) 11'IN'tII IN 1'//ls tr~.•.r 1.. Itw tir.rsr % W,r I rMr. •I 1 .1, 1,1 A... Ila...,wv 1.1 •... 1..r1 The ~ne/ic deltct Iw alptu-l-anlilrypsin derKKn1 pa1KMs GARY 1). FRIFDMAN, M.D. Stniae F plde.niologid, I)eparlmeM of Medical Methods Research. Kaiser FoundNion Research Institute. Oakluad, Cal. Characleristics of smokers and non smokers Twin re{istry feasibility study The relations bHween smokin/ rtsolives, personality .nd feelings I Cigarette scnoke e8ects o. ccr.ain apecls of ta1 lung metabolism JACQUES F. GIELEN, P1t D., Aulaawl Pro/e»o., JiAorory of fltdicaJ CAemisny. Toawofody .nd NyTiene, Institute of Pathology. University of Lie/le, Liege. Belgium. Cigarette smoke and ptdycyclic l.ydro- carhon metaholism in rat and mouse lung and kidney enelic and environmental faelors af- GERALD 1. Ol-FICH, M.D, Consrfranr in M.dicrnu, Researc\ L.hocalory for Alkrtlic Diseases, Mayo Cliwie, Rocttes- ter. Minn. Hypersensitivity to ami/cns (rom tobacco as a r factor in the puhokencsis of chronic hronchitis lecunA smoking hchavi-A LFONIDE GOLDSTEIN. D.St., Asao- The "chronie nicotine uale" and anaiely ciere pro/runr a/ rrychiarry. InslNute a behavioral and ekctroencephalo- for Mental Health Scienoes College of Bnphic analysis of induced and spon- Medicine A Dentistry of New Jersey. taneous hyperaclivaliun in ra/s Rutgers Medical Sct.od, Piscataway. Pllects of tobacco smoke and nicotine on coronary collateral blood b. JOHN W. GORROD. D.C.C., Lecturer in Tie metabolism of •'pyridines" in relation AiopA.rwr.ey, Ct.elsea College. Utsiter• to Ihe induction as/ ncoplaYic disease aity of London, London. MARU/T IIAM(>511. Pn.D., ResercA TUe eilecl of c/tarelle snroke on lung Au..r•iart. Department of PAysiolaRl meuMdism Ilnitiated unrkr Vid/l• B 1 Metabolic response to stress-tohanco and Mophysicr• (;eorpNoww Univer- smoke inleractions sity Schods of Medicine and Dearis• Relationship of non-MM phcnolypes and lung disease among intams The inheritance of the srnoklng habit Srud~rs on pepllde MrnA spttintilles. aerr.r s.lr and rnMbd/un (of hunnan leu- .,. a p..nr.vs . hnh a.t .mpluated in .Ir pab.yenr... nf pldm.rnary em t•1.. Yn.• try, Washington. D.C. PAUI. /tAM(>511, M D., Assoriarc rro- /esrrw of rAy,lolosy and OiopAysks, and M.dhire• (korgetown Uni.ersily Schnols of Medicine and DtrMislry. Washington. 1) C. NORMAN W. HFIMSTRA. Pst.D.. rro- /e,srn of l,ycAology: Dirretne, Hrman Facrar, L.borarary, Universi/y of South t)akota. VtrmlBww. HF.RRFRf B. HFRS('OWIT7, Pnt). MlunAadnRr. Arurt.nt lrn/esror of (itorgeluwn (Iniversily SchtNdsol Med- icine and 1knlrslry. Washington, 1)C. TAe eReat of smoking on the "smsll air wayi Fflects ../ s.ootms dep/rvation un rrsk taking Uchaviw BthavNNa/ e(Ietl\ rM rMM)nN,1f1s of t/ pnsll/e In smnt/ng FflecM o( cigarette smuke espl/sure o0 devell.pmtntal, eellldar and nlukcular aspetls of Ihe immune rtsponse 68 RO

0 PRINCIPAL INVF,3TICATOR OR 1N17TTUTION ALPHONSE 1. ING/ NIIU• Pn D, Aru.- ciarr rro/eisw o/ Ph..m...doRr. 1 he Albany Medical ( ouere of Union Uni- vcraily, Alhany. N. Y. (No.• so Fast Carolina University School of Mtdi- cine, (:reenvtlk. N. C.) HARRY 1.. IOA(-IIIM. M.D., ArrrndinA rNholoolur• Lenot Ilil/ Hospi/al; Clini- c4 Pru/nuar, o/ rarA.,lory, Columbia University Colkre of Physicians a Surreons, New Yurk. WILLIAM 1. 1tISKO- ht D, Aruwiare rro/e,usr of rbarrnac rnri. s; nirrrrw, Clinitd rArnw ni inetirr Laborarar y, Millard Fiilmore Hosoild, Bu/tab. EDWARD L. KLAIBFR, M.D, Srninr ScienNal• The Worcesler Foondalion for Esperimealal Biology. Inc. SMews• bwy. Masa. STl0 KULLANDFR, M D., rro/nn+r and CAairnran, nrpartmrnt of Obsrn- rira and cyn.r..forr. University of Lund. Land, Sweden. MICHAEL E. I.AMM, M D., Prn/rrfor of ruAolory, New York Uniaruty Medr cnl Center. New York. PAUL S. I.ARSON Pss D. H.n~ rrn/rs- aor oJ ril.rwwro~orr, Medical Cdkee of Virginiq, Richmond. IOSP.PH M. I.AUWERYNS, M D., Pw D., rro/ruar Ordinarirs and ('lrarr- ns.n. Depr(nsenr of rarAofory and Mkrosrorlc Anatomy. EsPerimental Laboratory of Pulmonary HrslopNAnl- ogy, K.tholiele Universikil /e l.curen• Lswets, Belsiwn. RICHARD A. LERNFR, M.D, Ara.hiaa MrnsArr, Scripps Clinic and Researrh Foundation. 1.0 lolla, (-al. IAY A. I.F.VY•'M D• Assistanr Clinlral Pro/rssor of Mrd.rfnr; RruarrA Asan cWr, ('anctr Research Inslilule, tini- vtrsil~ of California Sct•ool of Medi c.nc. San I-rancf.co CI.AYTON O 1(M)cl1 Pwp• M1). lh.u.nr. Pr..frrw.. of Mrdwrwr and rarA.H..ey. Unsers.ty of Srwlhern ( ab (orma School of Medrerne• Los Anrcles. PROIF.Cr TTT1.P Ac1uMs of carbon monoslde and tuhacco snwke on retinal rnel.lxdism anJ func- tion . Tlse immune respmae at the lumor site in lung carcinuma F/fecl of smokinR and its crssaion on drug disposition Cenlral nervous system adrencrelic func• Iioning and cigarette smoking Studies of a gonadal and central nervous system syndrome IhN diflcrawiales smoktrs from nonsmokers Infhtcnce of smoking on humtm Ioelal growth and pxilrtatal devcMpmenl, and un fihrinolysin in the bl.Nd of pregnant women. Accumuloliun of nicol,ne and/or damage to human pla- « nul and foelal lung tissues Immune mechanisms of mucous mem- branes Publication of Supplement 111 to Tolacro, If61 The neurnepithelial Mdies: their rde and slructure as iMraprlmonary newo tchemukeceplors in normal and rari- ous physiological. pharmacological and pathological condilions Studies on persistent vird infeclion Oncornaviral gene espressNUS in normal and maltrnant tissues Possible genetic delerminanls of chensical carcinorcnesis I fltals a./ fresh cirarctte smoke inhalation on the respral.>ry tracl of mice 70 I PRIN('IPAI. IN V FSi1GATOR OR IN.4TIII171ON 111-NRY 'f. I.YN( H• M.I).• Pr..feuor rmd ('banrnan. I)eparfinenr u/ rrrvrn- uvr MrJ..inr and Pwblir Ifrulrh, ('reiRMun tlniversily School of Medi- crne• (Hnaha, Neb. RI'(71NA1 /) G. MASON. M D., Psi D., /'orhulucur-in-C'hir/, Memorial Husp- lal, Pawlucket, R. 1. (:I:kA11) li. MCCLF.ARN, Pu.D., Di- rn rur, l.urlfnrr /.w RrAariwwal (;r- nrrres, /)rperfnrenr ../ Pr?r Ard.Ky, Uni- vtrsity of ('oMrado ScMal of Phar- macy, Buulder. HERBERT MCKENNIS, lr., Pr/.D.. Prn- /rssur of PArnwcolur). Medical Cd- kre of Virrinia. Vir~ nN CotnnKrw- weallh University. Ricfnsord. HANS MEIER. D.V.M.. Senior Srat Srl• rnNsr, The Jackson LaborNay, bar Ilarbor, Me. DUV MIC'HAFLI, Ps1.D.. ADiuanr rro- /ris..r of ANN Armiury and SrArr~ University of Californ Scbdia Medicine. S.n Francistw. MtCROB1OlOOICAL ASSOCIATES. INC., Belhesda, Md. /. ANDREW MITCHt91.L• Pw D., .Nsisr• anr rrofrs.w .rf Anar..my, Wayne Stale tlmvtrsny Schu..l of Medicine. Deuocr (7FOR0 R. NFtIRAT H, Pn 1) . A(i. r.r• analyrical Lob.MOrwy, Ilamlwrg, Wes/ (iermany. PROIE('T lT 1 LE Smoking history in famdics with low anJ high onser rnadcnces Aryl hydr.cathun hydrosylaK (AIIII/ cancer genetics I:Recls of nicotine on inlefaclions u/ p1alelNs and enJ.Nhclial cells //crc.Lly and tubacco rclalcd Lehavi.u in the m.wse Cunperaive studies aimed at Ihe .kvel opmcnl ul immun)...ays /w ni.W.nc and nicotine me/aMdues Orscorenesis in the rabbit: genetic sus « ptiM/ity, vertical transmisswn o/ rirus and environmenial influenccs Transplacental effects of niuosocom pounds in inbred atrains uf mice and rabbils F./tecls of cigarette smoke on pdmonary fiMublasls and collagen and iss rela lion so emphysema Development of a mcwse modcl sysuem (a genetic susceplibility and its relation ship to in riro lung catcim.senesis I)evebpnenl of in sin.o and in uu0 nsu.kl systems for the study of lung chemical suxepthiDty and carcimr genesis Smokc inhalation carcimtrenesis studies in mice lluman A/111 sludies Resear.h sefrNts in •uppltl nf MfNI prrycsls Isce 1 evy, lay A and Wang. 11 A study o/ Ihc tfttcts o/ nn.Nme an Rcu. In.n in /he ca/ wirh parraular fclrren.e lo uuplanlanurs and the t.n.r u/ un•rr ul parturdwn Nitrosamines in luhactu and ns snrokc uccurfence, lurmaln.n and aam/er 71 I

I PRINCIPAI. IN V FffTIGATOR PROIE("f TIT1.fB PRINCIPA 1. 1 N V iS19Cq?.DR PROIF.(T 11T1.F OR IN311TUT1ON OR INSTITIITI()r( KFNNF:TH PAI(iFN, Pul).. Drrnan, A Rcnclic Ies1 of /lucorunida~e in Mad- J SCRIPPS CI.INIC AND RESEARCH 6mmunological competencc and csemicr Drl.nrrrnrnr ..f Mrdrruler di.daFr, cr cancer FOUNDATIUN, La loBa. Cal. carcinoRenesis Health Re.carch. Inc. Roswell Park Division. BuOalo. CAR1. C. SFI.TZER. Pw.D., Honor.ry Constitutional studies relative to snwrlin RrsrarcA A»oriae, Peabody Musewn, and coronar y heart disease CAR1. W. P11 R(F. M.D., PN.D., As- Biolo/y of suppreuur T cells Harvard Univetsily, C.reEridge, Mass. a.M iarr P...frrr..r of P.rAoluRy. flat- • vard Medical School. Boston. CHARLES R. SHAW, M.D. Chief. Sre- Hyd.oe.rbon nsetaboliting enzymes an Nnn of Medical Grnrrks. M. D. Ar- lung ancer ILARI RANTASAIO, M-D.. Pro/rawn The f-innish Twin Registry derson IlosPi/aI and Trwlor Instilwe: and CA.ir.n.n. Dr~arrnreM o/ PuMic Pro rsso. of eiolop. The University NrdrA Science. llnrversily of Nelsin\i, of csas .1 Houslon, Houslon. llelsidi. NATIIAN 11. SLOANF. Pw.D Pro/rs- Flfec/ of 6enr..iulpyrene and derivalive RONAI D F RASMl1SSFN, Prr D, As- FRect of eoearcinogcns and hrmor po- , , sur of diocArnsis(ry. The l)niversit r on mammalian lung cells /IMMfr Rrcr.r.lr Phyric.f..Rirr, Ikparl- moters on DNA repair in mammalian Tennessee Center for the Hca11A mcm d Cnmmanily and Favironmcrw• cells aucepuhk lo chemical Iransfor- Z iences. Memphis. d Medicine. University of Calilornia malion Colkge of Medicine. Irvine. TIMOTHY I. RFOAN, M.D. Profrssrn• of Medicine. Dirrcror. Diriswn of Crdio.ucrlrr Drsrsrs. Cdkgc of Medicine and Ikntiary of New Jersey. New /crscy Medical School. Newark. LYNNE M RFID, M D, Wrdb.wh Pro- frcwrr of Pnh.rlnRy. Harvard Medical School. Boslon: ('h.nnren. Dry..rnnrnr of P.rAoloty, Children's Ilnsptal Mediul Center. Bwlon. DANIFI. B. RIFKIN. P11.D, Auisranr Pro/s.or of Chrmind AIo1oFr, lhe Rockefeller Universily, New Yorl. CHARLES L. ROSE. Ps(.D. Clink Dirrc- (or; Director. Ncrns.ri.r Aginj Srrdy, Ve(era(us Adminirraliow Oulpatirn( Ciil.ie, Boro0. JOHN A. ROSECRANS. Ps( D.. Asro- cwe Pro/rrsor of PArnrocolory. Medi- cal CdkRe of Virdieia, Richmond. UNA S. RYAN. Pq.D. Senior ScirnHrr, Papsnkolaou Cancer Research Ins1i- lule, Mlam/: AI.iHant PlojralfM of Medicine. llniversily of Miami School of Medicine. Miami. Fla. f1. V. RAMA SASTRY. D Sc.. Prt D.. Professor of Ph.rm.c.>losr. Vander- bill Univeruty School of Medicine. Nashville. 1 cnn IAKOB S('llMtl)T, MD. Pwl). As. rnrrnr Profrn.w ../ llurhrmuhr. Ih vision of 1twJupcal Scrences. Slale t/mver.oy of New Yor1 at Slony Brool, Sluny flruu\ Variables affecting the cardiovascular re- sponses lo ehrunie smoking The eflecls of irritalion and drugs on luway epilhelN.m An e.perrmemal sludy of inechaniuns ProMCascs produced by mammalian lung lnsue A smoking research program in the Nor- malive Aging Study State de~c nden/ properties of nicotine- relaleJ compsunds i LOUIS A. SOIOFF. M.D., Bl.ncbr P. Lrry DisdngrrisArd Ser.ke Pro/rssor; Professor of Medicine. Dirrcror. Rr- sr.rcA CJjii/ l..borMory, Temple Uni- versilr Heahtt Sciewas Cenler, Phila- dclphu. DAVID W. TALMAOB, M.D.. Direcror, Webb-Warind Jnstirwr, UniversNy of Colorado Medical CeMer. Denver. MARC D. TIIAMBS. M.D. Srnlo. Re- ar.rcA Frlfow, M•~o Clink .Id Folal- dalioa. Rochester. Mi(tw. Role of lecilhin: cholesterol acyl Iran• Ierase (I.CAT) in chdesterd metal. lism in heall6, disease sad dunn smoking Purification of .n antibody poducliu to keitsin: eholeslerd acyllraasfers. Tlse role of maeropAye-induad (acto, ii: cancer immunity Cardiac mechanoreccploes and reain rr lease JAMES TRAVIS. PN.D.. Aaociut ho- /rssor of AlocArndsuP. University of Oeorgia. AIheM. OF.RAI D M. TURINO. M.D.. Pro%ssqr of Mrdirlnr, Columbia Uwheniy Cd- kge of Physicians A Swgeaws, New Yolft. Biochemislry of chronic obstructive 1un disease Chemical basis of liswe destruction r. obstructive lung disease D. M. TURNER. Pn.D., Nrd, D.p.r(- Buccal ahsorpion of nicotine In th Endocrine functions of Ihe lungs . nrrn( of Drug Mrt.bolism ond Ilo- anestheliaed ca1 Hark(on 1 aMralories P.u- cArmirrry rc.pe, Ild, Ilarsovalc, Yortsbire, Fq• land. FMII. R. UNANUI; M.D.. M.IIInrJrelr Physiopalhology of normal •nd a./i.arr Influence of nicotine on the rekase of Profnsor o/ Jmwrrw~.~ Aolady. Har- maerophases acelylcholine in the human placenta and vard Medical School, Boslon. its implications on the fetal growth Central nicotinic receplors UNION CARBIDE CORPORATION, Nuclear 1)ivision. Oak Ridge. Tenn. Ch.rrc/erinlion of ani.n.l esposure s)• lerns ('haraclerirala+n of ammd inhalation es pssure devices UusimNry s/udics 72 71

! PRINCIPAI. IN V FTi7GATOR OR tN1i1-IIZliION UNIVERSITY OF SAN FRAN('ISC'O. San Francisco. STEPHEN F. VATNFR. M D, Arsoclatr •Nkoline induced refka coronary vasodi- Pro/es,or of Medicine. Har.ad Medi- lalion cal School; As...cJerr In Medicine. Peter Benr Brigham Hoqital, Boalon. /RANISLAV VIDIC. D.S., Professor o/ Anaron+y- Otorgesown University Schools o1 Medicine and Denlislry, WasNsR(ow, DC. (Sea Ilrnosk M.) IRENE Y. WANG. Pu.D., Asslsraws rro- /rs,or of Bask and Cfinkvl hnnruwd- ory and Mkroeloloty! Medical Unl- .ersNy of South Cudrn~ Cbarleston. PR(I1F.(T TITLE A study of squamous cell earcinoma in mouse lungs F.aposure of mice to ashestos nd ciga- «lle smoke Nkotine-Induced reBex activation Ct.ARFNCF. M. AORESS, M.D., Asso- R. FRFOERICK BECKFR, PnD, Mro- uatr Chnr.al Pro/rstor of Mrdicint. .urr Professor of Anatomy and Owrc- University of California Medical Cen- fur, [.aAwaro.y of Prnnarol Strre.r, The elfect of cigarette smoke on lung ler, 1-os Angeles. Dut,e University Medical ('enter, Dur- n.e1alolism ham. N. C. Genetic diRerences in the In vitro metabo- tism o( ehemicd carcinogens by buman and mouse lioues Completed Projects Following is a Gst of the principal investigators, or institutions, of projects that have been completed prior to the period covered in this Repxt. Several of the individuals named are deceased. The titlcs and affilia- tions listed are those in eQect at the time the work was completed. AN t IIONY A. At MANESE, Psl D. D/- rnaw of L.Ao.arorirs, lhe Burtte Re- habilitalion Cewer, Wtsil. Mai., N.Y. ANTHONY P. AMAROSE, Pw.D. In- .rrarro. In Obtttric, ond Gynerolofy. The Albany Medie.l College of Unio. University, Albany. N.Y. PrnJrs- LEE W. WAT'TY.NBERO M D. Inhibition of eareino~cnesis by hcn:yliso- i , , ,w of f arhoro(y University of Min- Ihiocyanate and relaled compounds F. T. ANOELAKOS, M.D., Pw.D., Pro. . Minneapolis. nesota Medical School /es,or of rhysiolop, Boslon University , Scbool of Medicine. sonors. PsMO sioral- GEORGE WFINRAUM l.ung proreinase:antiprotinase balann . , rn/monary Disease Section rnNsr AI- and Ihe eRecl of ciprenle smole on tbb D. MURRAY ANOEVINE M.D., Uni- , . Phita- bert Einslein Medical Center inler acl ion .ersily of Wisconsin School of Medi- . delp6i.. cine. Madison. Prn/rs- P>t D THOMASrC WPSTFAI 1 Action of nicotine on peripherd and een• STEPHEN M. AYRES, M.D_ Dbecto., . , . . University of Vir- ,ar of rllarrnacolocf Irsl neurons in animals chronically et- ('ardropu/wronary "borarory, Saint . ginia School of Medkine, Chulol(es- posed to nicotine Vincem's Hospital, New York. tilk . OSCAR 1. BALCHUM, PN D., Hastlwp IAMES A. Wll.l., O.V M.. PN D. Pro- /r,sor and Chairnsan. Deprrnstnr of Ynrrlnr~ Sc/rwrr, University of Wis- oonsirh Madiaon. OEOROE WOLF, D. Pfsn., Professor of I'Ay,loforJcd CArmbrry, Drp.rrwrrnt of Nutrition and Food Srknre, Masv- clwaclta Inslitwe of Technology. Cam- Midge, Mar. Morphologic and functional corretations of the APUD «lls of 1he lung The eRect of vitamin A on glyco}rolein synthesis in normal and pecanarow respiratory epithelium F.Red of nrcinoRens on tlycoproletn aynthesis Professor of Mtdiclnt. Uni.ersity of Soulhern Califorw School of Medl- eine, Los Angeles. FREDERIK B. BANG. M.D., rr essor and CAairrnan. DrpartnxaN of asho- 6wlos~. The /oMs HoPliwf University School of Hysiene .M Public Health. Baltimore. KO// YOSHINAOA,.ny,Pss D. Asu~clane Pro/t,sor of Anaroy. La"aro.y of Hrnwn RrprodwHnw and Rrp..drr- M.e slolory. Hanard Medkd School. BoMon. FRects of nicotine on pregnan:y 74 A. CLIFFORD BAROER, M.D., Robert Hrnrr P/rlerr Professor of Physiol- o/ly.'larrard Medical Schod, Boston. BRODA A. BARNES. M O, Pn D., lro- Irssor (Asdiuel of rApioloff, Cdo- rado State Unisersity, Fort Collins. FREDERICK W. BARNES. la.. MD.. Associate Pro/rssor of Mrdicinr, Tle /uhns Ilopsins llnlsersily School of Medicine. balrimore. T. C. BARNFS, D St., Rt,rarcA Srirn- Nir, Philadelphia State Hospital. Phila- dclphia. RAt PII S. B1-CKFR. Pe t) , P.ufrsror of ( hrmnrry, Uni.ersny of Ilouslw. Houston BFNIAMIN BFLL M 1), Unruar F.nr- rrrrr.r, N.wnwrirr AKrnP Srudy, Veln ans Adminiuralion (hrrp.licM Clinrc. Boston. SAMUEL BfLLF.T, M D., Dverro., Di- ririon of Cardrolnrr, Philadelphia General Hospild. Phdadelphia. BARUI BENACERRAF. M D., Fa&yan Professor and Chanrnan. Drprtwunr al I.rAulup, Ilar.ard Medical School. Iloston. 1O11N A. BFVAN, M D., Professor of rh.nnwroloRy, llnisersNy of Califa nia School of Medicine. t.os A.yeks BUDt10EV BIIAOAT, Pss O., Professor of Physrul..Ry. Saint t ouis Univeruly Scbool of Medicine. St. l.ouis. CESARE BIANCIFIORI, M D, Dirbbn of C'an.er RrsrarrA, University ol Perusia, Perudia, Italy. HYLAN A. BICKFRMAN, M D.. Auirr- ont Pro/rn'w of Mrdreinr, and AI. VAN 1.. BARA('/l, M D, Cunrdaer rw Meda•rnr, ('olumbia University ('oi kae of Physicians i Sur{cons; (3dd water Memorial Hospital. New York 810 RFSF ARC11 ('ONStII l AN I S, IN('., Camhridge, M.u. R10 RF'SFARC'H INS111111E, INC. ('amhridge. Mass FRfU G. B(K'K, Pa 1). Arn,aurr ('an- rrr Rrvarrh S.urnrru. Aialogird Sra u..n. Roswell ParL Memorul Insurmr. Sprsnil.rlk, N Y HFRMAN V Bt)1 NIt1- Pn D. Hred. I)rpartnrrnrof (hr.rou.r.nd Ro.. hrarrsrry. Sppndlewp Rescuih ( enter. 192 mgrun. Ky. 7s i

I I I JAMES F. BONNER. Pw D, Pro/r»or of Riolury. C.lifornia lnstnule of Tecbnology, Pasadena. WAI.TER M. B(X)KER, PN.D, Pro/n- sw and Need. Drpr tmrnr of Phrma- colory, Howard University. W.sbing- lon, O C. TOM O. BOWFRY, Pw.D., RrurrA Pro/nsor, PnrNide Residwe (.aAora. rw' North C.rolina SL(e Collere, Raki.b. OEOFFREY L. BRINKMAN. M D., Ar sorrare Pro/euw of Mrdreinr, W.yne Sute l)wiversisy School of Medicwre, lk/roil. ROBERT F. BR(X)KS, Pw D_ Aasoclare Pro/ruor of Parholopy' Unirctsily of Orepon Medkal Sckod. Portland. BARBARA B. BROWN. Pn D, Chief. Esperbwrnal PrqcAAstry, Vetetens Ad- niiniwra(iow Ilospital, Scprlvcda- Cal. RAYMOND R BROWN, PN D. Pro/rs- sor of Clintral ()wcololy, lJni.ersity of Wiseonsin Medreal School. Madison )OSFF BROZFK. PnD, Profrrror end CAairn.an. Departn.rnr o~Pr t. A.dnt r. Lebig! UsirerlMy. Betble m. Pa SUP. BUCKINOHAM, M D, Atduawt Pro/essw of Prdutncs. Columbia Um- versity Cdkge of Pby.icians i Sur- teons. New York. §ENIAMIN BURROWS. M D., A»o- cdan Pro/es.aor of Medklne. Universi(y 41 Chicago. Cbic.p. E. M. NUTT, M D, CA/e/ PatAolqla. l.oe A.ge/es CouMy Oeweral Hoapr(d, Los Angeles. RICHARD U. BYFRRUM. Psr D, Pro- rssor of CMn.lsrry, Michigan Stale niversity. Eew lansi.It. SISTER M. EMILY CAHII.L, Prt D., CAaknsan, Drprrn.ent of Chemistry. Regis Cdkde. Weroq M.u. BRl/CE F. CAMF.RON, M D. Pa.D, 1(ow.rd 1(u~hes InstitWC. Uni.erurr of Miaml Scbw.l of Medicine. Miamr. FIa W1111AM FI ('ARNFS. M D, l/nirct- wry of lhab ('olkle of Mcdicrne. Srdl I ake ( ny MARCUS N. CARROLI., )e., Pw.D., Chief. /)irision of Pharm.rrolury. The Bruokdale Ilospiul C'cnmcr, Brooklyn, N Y. WILI IAM ALVIN CAR (FR, M.D. .Irsiuant PruJrssur of Mrdwine and MN.oAiolary. The /obm Hopkins Uni- versi(y School of Medicine, ballirnwe. I EOPOLD R. CERF.CFDO. Prr.D., Pro- ,nsw .// einchrmiurry aNd Nrnition. Unirersi(y of Puerto Rico School of MedKine. S.n lue.. C1111 DRFNS H(>SPITAI. OF LOS AN- OFLFS, l.os Angeks SANFORD CHOD()SH, M 1), Au/sranr Prolrssor of Mrdrelne, l ufls Unlver. 2Ay School of Mediune, Bos/on. NAII'ER M. CHOPRA. Pw.D., Pro%s- sor o/ CArnsisrry, Nor1b Carolina As- ticulturel and Technical Sute Univer- sity, (;reensboro. WIl1.lAM O. CLARK. Pw D, Dkecror, Psychophrnracol..ry Rrsrrch laew.- rwy, Veterans Admimrraion Horpi(d, Sepulveda. Cd. IIANS T CI ARKF. 1) Sc. Pro/rssw of P..r Annurrr, ('ulumbu Unirnsdy < ullcge of Physiauns A 1uteeeons, New Y(«k /AY 1) ('OFFMAN. M D, Section llrod, Prrrphrral 1'ur.ler Drporrmrnr, l/mrasUy Hosppul, Boston. DANIEL COHEN. D.V.M., M.P.H, As- stsranr Pro/nsw of Ynrrinr' Epi- drnriolo`y and PrAlrc Health. Univer- siry of Penssylv.ni. School of Veter- inary Medicine. PAiladelphia. JULIUS 11. COMR(E, /.., M.D., DMec- rw. Cwdiorascrlar Resrarrh Insritn(e, Universi(' of California Medicd Ces- kt. San Francisco. DF.AN M. CONNORS, M1). Auorlare Drrrrtur, Drp.r rn.rnr of I aAoratory Mrdicinr, St. Maty's Hospital, Madisoa, Wis. P1111 IP COOPFR, A/ D. Clinkal Pre- /rnor of Surgery and Dirrcrw. Srrb ral I.aAwaawy of Crllrlar Physiolory. Alberl Einstein College of Medicine of YesMra Unirersity; Chir/, SwtIral Sr+rke. Veterans Admlmattauon Fto.- paal, The Brons, N. Y. 1O11N F. CRAIONEAD. M D, Pro%e- s..r of ParAolory. University of Ver- mom College of Medicine. Burlington. 7A i ROBERT L. CRAIN. PN D., Assistant Professor of Socioloty. Univcrsily of Chicago, Chicago. T. TIMOTNY CROCKER. M.D.. ProJrs- s..r of Medicinr. University of Cali- lorni. College of Medicine. Irvine. CECIL E. CROSS. R~uarch Drprrmen(, St. Joseph Hospi(. Burbank. C.1. AI.BERT DAMON. M.D. Pw.D., Ln- rwrr on Anrheopolorir: RrsrrcA Asso- rcarr in Medtcd Anthropology. Pe.- body Muscum, H.rvard Utriveraily, Cambr/dge, Mass. 111OMAS R. DAWBER. M.Dy Associarr Pru/rrsw of Medkiwe, Boeloot Unlver- u(y School of Medicine, Boston. R. F. DAWSON. Pw.D. Pro/es.or of eo(- rry. Columbia Uni.ersily, New York. /OHN P. DELANEY. M.D., Pe.D., As- sociarr Pro/ersw of Srrrry. Uwiver,py of Minneso(., Mis.eapolis. ANDREW S. DIBNER, Pw.D, E+ec- rNrr, Ppcho-Rese.rcA, The Age Cen- scr of New Endlrmd, Iwe. Boston. EDWARD F. DOMINO. M.D, Pro/es- u» of Phrn.arolosy. Unlve.si(y of Michigan. Ann Arbor. RALPH L. DORFMAN, P((D., Director of Laboratories. Worcet(er Foundation for EapetimeMal Biology. SMewabwy, Mass. JAMES 1. DYAR, Pw.D.. AstluaM Pro- /rssor o/ alolop, Bellatnine College. Louisville, Ky. • RICIIARD H. flARLE, M.D., Chk/, Prlmonry Function l.8orarwy: As- siuant Pro/nsw of Medk(nr. Ueieer- sity of Cbica.o. Cbicaeo. /OIIN W. F•CKSiE1N MD.. Assistant Pro/rsuor o/ Inrrrnaj Mrdlcrnr, Sure Unircrsny of Iowa College of Medi- eune. Iowa ('i1y. BFRTRAM FICHFI-, D.DS. Dirr.r.a. Instlcrre of Sromatnlorkal Rrsrr.h, Science Rcsources Moundsion. Water- lown, Mass HYMAN ENOELBERO. M D., Anrnd- !nr Physlci.n, Cedars of Lebanon Hos- pital. Los Angeks. I CAKLION K. FRICKSUN, Pu D, ,(s. s..c.atr Pru/rss.a of Pharnm.o/..st and Tostcolutf, lhe llmreraNy of Kansar School of Pharmacy. Lawrcncc. H1:NRY 1. ESBFR, Pit D., Resrorch Tm- mrnulorur- Mason Research Institute. Worces(er, Mass. 1()IIN R. FSTERLY. M D, Asswlare Pro/essw o! Parholory, lJnivcrsiry of ('hic.go Prpzker School of Medicine. Chicago HANS l.. FALK, Prs D., Adjunrr ArNK/ are Professor o/ Pothol..ry. Universuy of Southern ('alrfrwma Scho..l of Medicine. Los Angeles. DANA L. FARNSWORTH. M.D., Nrnry A. Oliver Professor of Hygiene and Drrrcrur, Unirrrsrty IlydrA Srrrars, Harvard Uwiversily, Carnbridiie, Mau. FRANK C. FFR(IUSON, 1.., M D., Chairman, Dep.rtmrnr of Phrma,ol olly, The Albany Medical College ol ltnion Univetsily, Albany. N.Y. THEODORE N. FINL.EY, M D, Darc- rw. Pulnronary Rrsrr.h L.Auruory. MoLn( Ziow Hospual, San Francrs.o. W/1.11AM 1. FISIIBEIN, Ml), ('A/rl of Epidrn.idory, Cbkaro Bo.rd o( I/eallb, Chicago. EDWIN R. FISIIFR. M 1), Dnrctor ol I aAo.arwies, Sbadyside Ilospilal, Pro rssw ol Patbnlory, l/niscssity ol iushursib School of Medurne. Peu lwreb. RUSSF.LI. S. FIS/IFR, M D, llnirtrsil/ of Maryland School of Medicine. R.i Umore. 8. L FRFFDI ANI)1 R. M D.. Dorrto, a/ C'an.rr Rrrrrrh, M.wnl 7ion l/rw piul and Medical ('enur. San Fran C/Ko I:RFDFRI(' A. FRFNCI/, A R, Purw rw of ('on.rr CArmothn.py Rrvar.A Mount 7.ion Hospil.l and Medica$ ('enter, San Francisco. lA('K FRFI/Nl), M D, Auhuwr Pro ,rINM ol Pharnrar.dorr. Medical ('0) +qe of Virrmia. Richmwd. (;11 BFRf H FRIFUFII., M 1), ('AIN of Path.d.r`' SI. Vincent /lospul Worcesqer, Man. 77

I I 11. IIUG11 FUDENBERO. M.D, Pro/rt- ,or of Mrdicinr. University of ('ali- fornia Medical Cenler, Ssn Francisco; Professor of A.rrrrldo`r and lmnrr- nolo[f. UniversilT of Cal~fornia. Berke- ky. ARTHUR FURST. PN D, Dirrrto., fn- uitwr o/ Chemical eiolo[y, University of San rancisco, San Francisco. . MURRAY B. OARDNER, MD Auo- ti.tr Professor of Pahotoq. Z1.iver- sily of SoaAerw California 3cKool ef Medicine. Los Angeks. OEOROE O. OF.Y, M.D. Dkertor, Fin- nry-Howrfl C.ncer RrsrrcA l.Aoro. torf: Associate Pro/ruor of Sr~rr~~ T!t loMn HoPtM. Universily Scbol of Mcdicine, B.Rirwa... TTIOMAS M. OOCKLr, M D.. Assoclar Professor of Prrrrntirr Mrdklnr and Cen.wsrnlty HrdtA, Sc1on Hdl Col• klge of Medicine and Den(isrry, Jersey Cwy. N. 1. DAVID M. OOI.DENBERO, Sc.D., M D., Auociatr Professor of P.rhol- otf. Temple University Hcdl\ Sci- t.ees Cemer, PWadelPAia. PAUI. GOI.ONABER. D D S, Anoclarr Pro/ruor oJ Prriodontolo[y, Ilsrvard School of Dental Medreine. Boston IRA OORP M.D.. Proles,or o/ P.rhot- M Boaon Usiversily School of Medi- ; Chief of Loboratory, Servkr, ve(er.n. Admi.iMruio./lospild. WeN Roapw7, Mass. OERTRUDP. Y. GOT[SCHALL, P)M D" Assisunr Professor o/ Slorhrnsiaay. Columbia University College of PRysi- dane A Swgeons, New Yort. A. CLARK ORIFFIN, Pu.D_ Head. Department of Aiochrmi,tr), M. D. A.dersow Hospital and Tanwr Insli• 1We, Urrversily of Teuas Medical Center. HoYsloll. ARTHUR L. OROSS, M S., Srnlor dio- chrmiu, Southwest Research Institute. San Anlonio, Tet. MORTON 1. (iROSSMAN, M D, PH D, Associate ('Irnka/ Pro/rnsor of Mrdl- ciwr. Universiry of California Mrdksl (•enltr• I o,. An[tlts /'ull t. IARI ( /iM1-/1/11 M1t ... . . r,....r ( .•.r r... ...•l ,.r.• l •n.... .., ..l '.n.. .1. ... .....t ..... ~ ~..'../ a.d..n. P..~.~a~re.. IOSEPH J. GUARNER.1, Pa.D., Artrnd- m[ Microblolu[ut; Director. Mitrobi- olo[y L.bo.arorir,, Long Island lew- i+A-llillside Medical Center. (lseerq Hospilal Center ARilialion, lamaic., N.Y. FRANK E. OUTHRI[:, Pr( D., Pro/es- soe of Entomology. North Carolina S1ale College. RakigA. H. B. HAAO, M.D.. Professor of PAr- wacolo[P. Medical CalkBe of VirRida. R icAmond. F. 1. HADDY, M D., h( D. Professor .nd CA.irns.n, Drrwt.nrnt of rhr,i- dn[7. University of (1klaioma Medi:al Cen(er. Otlasorna City. IOSEPN H. HAFKEtlSC/11P.L, M.D. DMector, Crdiopnln.onry Unit. 71e Lantenw Ho+p(al' A»ociarr tn Mrdl- cba. UniversNy of ~eensylvania School of Medicine. Plilade4DAia. BERNARD HANE PeD., Professor of Health Scirncr, C ifunia State Uaiver- sily. Nortsridge. RICHARD 1. HAVEL. M.D.. Assistant Profrssor of Mrd.cinr. Universil of California Medical Center. San Frraa- cisco. HERBERT R. HAWTHORNE, M.D.. , Charrm.n, Drp.rtmrnt of Su.~rry Universn~ of Penmylvania GradrNe School ol Medkine, P6iladelrAia. JOHN A. HAYFS, M.D. Associate Pathologist. Mallory Institute of Pa• tKoloRr. Boslo. City Hospital. Boslo.. CLARK W. HEATH. M.D, Professor of Mrdicinc .nd Dirtctor of Health Srrr- icrs, T.lls University. Med(ord, M.r. PAULINE IIF.ILP.R, Pw.D. Research A,rociate in (-yroto(~ and CytocArml,- try. San Francisco Insliluwe of Mcdicsi SCKncel, San Francisco. I.AWRF.NCP. L. IIPSTF.R, la., M.D.. Pro/nuir and Chalrm.n. Department of l)hstrtrnc, and Gynetc.lo[y. Medied ('rdkge of South Carolina, Csaskslon. 1NNI (IIR11% 111)1F. MI). Pr(/), /'..•h...w .e.l (ha..n1on, U.tldan of P... A...u.. Nr.re.. h Medecrl Collegie ..f % Rnlunund 18 I i i RUSSEI L L. HOLMAN• M.D., Louisi- sna Slale Univenily Scfaool of Medi- cine, New Orkasu. OLE A. HOLTERMANN, M.D.. Rr- ,eoch Scirntitt, Lc.bwnd Labo.aor). Umversily of Npre Dame, No(rc Dame. Ind. FREDDY HOMBUROER, M.D.. Presi- dent and Dirrctor, Bio-ResearcA In•d- luwe. Inc.. Cambridele. Mass. ROBFRT W. HULI. P(( D., Pro/ruar of Arolo[ic.t Sckncrs. Florida S1a(e University. TaRallasset. 11 r RFSEARCH INSTTTUTF Chkago. GEORGE IACOMSON, M.D.. Professor and Hrad. Dri*rtnsrnl of R.diolo[r. Universilr of Southern Californra School ol Medici.c, Los Aryeles. JERRY HART IACOBSON, M.D. D!- rrttor. Divi,iow of Etrcwo~Ayslolo[~. New York Eye awd Ear Infirmary. New York. lU1 lUS H. IACOB50N 11, M.D.. Asso- CIItr Professor o/Srgrry, and Dlrrrtnr of Surgical Rrsrrrh, University of Vermont Cdlqe o[ Medicine. Ilw- lin[lon. MURRAY E. JARVIK. Pw.D., Auoriatr Professor o/ PArnweolo[l, Albert Fin- aein Cdkge of Medicine of Yeshiva UniversiNy, T1e Browt, N. Y. OSWALD R. IONES. MD, S/. LnRe. Hospi(al, New York. ANDREW A. KANDUTSCH, Pw.D., Srae Scientist. Tle Jackson Labora- lory. Bar Ilarbor, Me. ARNOLD R. KAPLAN. P(s.D. Dker- tor, Laboratory of Mrdlcal Grnrtics, ('kveland Psychiatric Institute and Hosp(al. Cleveland. A)-r A1.1 AH KAPPAS. M 1). Pro/rs,..r and Senior PAyski.n. The Rockefeller Umver.iry, New Yar\. 11RATCH KASPARIAN. M D, Assist- ant Dirrctor, Crliswurl.r L.Aora- to.r: /nstructor in Alydkine. Ha6ne- msnn Medical Colkgge and HosPilsl. Phd.delPAia. 1:1.1H1/ KAT7 PND. Auo.latr ProJer- tor of So.folo[l. University d(-Ai- c.lo, ( AK.go. SHIRI FY I KAl1FFMAN, Mlt, /rsvn ..f PoN.oloRy. Stale Un.vr of New York Downstate Mcdaal Ier, Brooklyn. ANCF.1. KFYS. Pn D. Dirr.r.N• t oi tary of Phvsiolo[iral Nt[rrnr, 11isily of Minnesota School of P 1leshh, Minneapplis. IOSEPII B. KIRSNI:R, MD• P.o/ of Mrdirine. Univenny of ('A. School of Medicine. C6ica[o. JEROME KI FINERMAN, M D• 1. [)rviuon of Parhol..[l Rruar.h Clniut P.tA.do[ , SI 1 u1i s llo.+ ('kveland; Pro~rr..r of hth.. (-ase Weslern Reserve Univ. School of Medicine. ( kvcland. PETFR H. KNAPP. M D.. Rr,. Professor of Psychiaoy. Boslon versi(y Scsool of Medicine. Baslon KENNETH P. KNUDTSON, MD, versily of Washington School of A cine. Seattle. ALVIN 1. KOSAK. PnD.• A.sn ' Pr../rssor of ('hrmiurr, New Umversily, New Yock. ROBFRT A. KI111N, MI). A.u. Pr.r/rr4N, DItlllon .11 NeY.1..u.. New /ersey State College ol Medi Jersey City. MARVIN KUSI'HNFR. M/). Yor1 University McdKat Cenlrr. York. CHARI.FS W. I..BFI 1 E. Pw D..( .nr Professor of F.evironmen.al [.rnr. le/lerum Med.cal (olle[e. P delphu. AARON I I AI)MAN, PN D. P.ol• and l'haonwn, I1ry.arrmrM .ul ' omy• Ihe l/nrvrrssly of Ntr- hl. SaMwd of Mtdn.ne, All.o+ary..t 111OMAS C I AIPPI Y• M I), P, u.r o/ Perh.d.•[r. N.nrAweslt.n versny Medical S(b.wl. Clic.[•$ R(XIFR K I i1RtiON, M 1). ('Al. AlrJ...nr• I rt.no (ounly Il..y F.csnn, (-.1 (:I/SI AVi? A I A1/RPN71 M I) .( of Mrdu.nr. SI V.nccnr Ilo.y Worcesler. M..s 79

El)WARD I FF.TF, P)(.D., D Sc., Pro/en- sor of Chrmiroy. University of Minne- sola, Minneapolis. CECILE I FtK'HTENBFRGER. h(.D., Neod. DrP.rrw.rwr of Clrochrmisury, Swiss Inslaule for Eaperwnental Can- eer Research, Lausanne, Swirnerland. AVF.RILL A. LIFBOW, M.D.. CAdrl) man, Department of Poahology. YsJe llnives.ily Sc" of Mediciwe, New Haven, Conn. F.STFN O. LINDSFTH. M.D_ Psr.D., S(. /oseRlr's HosPieal Research Laboratory. S1. hwl, Minn ROt/F.RT 11. LINNELL, Pn D Assoc4 arr Pro/runr of CArwdsrry, dniver.iry of Verwwd, Burlingto.. HERBERT L. LOMBARD. M.D.. M.P.H., A/ili.rr, Cancer Rtserch fn- aiwrr New P.nRla.d Deaooneo Hos- Pi(al. ioaor.. l. P. LONO, Pit D, Professor of Ih.r- w.oro/oly. State Univenir' of (owa ColkRe of Medicine, lowa Ci1y. DONALD B. 1 OURIA, M D, Artociarr Pro/rssor of M.dkdwe, Cornell Univer- si(y Medical ('olkge, New Yo.l. KENNETH MFRRII.I. LYNCH. M D., Sc D_ Ll .D , Pro/ranr Fn.rrirus oj PNAolop and CAoncrll.w, Medical College of Sou(\ Carolina. Clukao.. 1NPS MANDL, PN D., Assisranr ProJrs- aer o/ llorhrmistry, Columbia Unirer- sky (`dk~e of Physki.ns • SurBeans. Ne. Yor~. JOHN H. MANHOLD, /.., D.M-D., lro/rasor .wd Director. DrParrmrnr nl PaA y owd Orol DiaRnosir, New lersey College of Medicine and Dcn- lissry. lersey City. DAVID E. MANN, PN.D. Assorlarr Pro%aser of PArws.colooly, Temple University School of P1arn.acy. Pfila- drclpiia. 101114 P. MANOS. M.D., lwrrrrrcror In Ylroloty and MoctrrJofogy, Medical College of Sowls Cardina, Cbarlesron. ('HRIST[)P/1FR M MARIIN, Ml), Anlurv Profnwr o/ Mrdw.we .wd D..rera+. D..u...+ of /w/rerrwr Das- earrr. Sernw /IJ/ ('oMep of Medicine. Jersey ('uy MAS()N RFSF.ARCH INSTITUTE, Wo.cesler, Mass. IK)NAI D/. MASSARO. M D., Anod- nrr PrnJrfsor of Mrdirine, George Washington Universily School of Medicine, Washinglon, D. C. CHARI FS McARTHUR. Pn.D., Psy- cho/oriV, University Nrnlrh Srn•ices, Harvard University. CamArWge. Mass. CHARI FS B. McCANT3, Pq D.. A»o- clarr Professor of $0118. North Caro- lina SIa1e College School of Apicvl- lure. Raleigh. lIF.NRY C. McUll.l., la, M D, AcNnd Hrad, Drporrmrnt of Pathology, I our- dana SIaMe Univenily School d Mcdi- eine, New Orkans. HFNRY D. McINTOSH. M D., Pro/rs- sor of Medicine and Direrror. Crdio- rasrr/r l.Ahororo.y, Dulee University Medical Center. Durham. N. C. FORDE A. McIVFR, M D., Associate Pro/r»or of Pathology. Medical Col- lege of South Carolina. ('harksron. EDWARD McKP.P M D., Pro/euor and Acting Chairman, Drparrmrnl of Pathology. Medical College of South ('arolina, Chatkslon. KF.1 l.Y T. McKF.F, M 1)., Associate ProJessar of Medicine. Medical Col- kAe of South Carolina. Cirarkslon. VICTOR A. McKUSICK, M D., Pro%s- sor of Alediclnr. The lohns Hoplins University School of Medicine, bahi nsore. ROSS L. McLF.AN, M.D.. Associate Professor of Mrdkine, e+nay Uaiver- sily School of Medicine. Allan/a. W11.11AM F. McNARY, in., PN.D., As- socurr Pro rssor of Awahrmy, Boslow University bd of Medicine, Boslo.. NEAL L. McNIVEN, PN.D., The Wor- cestcr Foundation for EaOerir.ewlJ Biology. SArewsbury. Mass. IUI IA MEYER. Pw D., .tss.rcl.rr P.e- frsrnw of or.l Pashnloly. University of Illonors College of Deml.rsy, Chicago. BFRNARD 1. MII t P.R, M D., Assistant lr.•/r+r.r n/ Anatomy. /efferson Medi- cal C'ollege, Philadelphia. e0 I JAMES G. MILLF.R, M.D., Pw.D, Pro- /rssor of Psychiatr) and Psychology; Dirrcr.w, Mrnral HeabA RrurcA !n- srirrrr, University of Michigan. Ann Arbur. CHARI.FS MITTMAN, M.D., Dlrrcror, Department of Rrspir.rory Dise.us, City of Hope Nnliond Medical Cew- Icr, Duarle, Cal. HUGH MONTGOMERY. M D_ Asso- ciarr lrofrrsor of Mrdklnr. University of Pennsylvania Scllool of Medicine. PIn1aSelohia. P O'R. MONiYiOMFRY, /._ M O., Pro/rssor of Parholog', University of lesas Sourhweslerw Medical School, I>rllas. GEORGE E. MOORE, M.O_ Pa.D., Di- rrrta., RosweM Park Mawotid Insll- tule, BuRalo, N. Y. KENNETH M. MOSER. M.D.. Assistant Pro/ersor of Medicine. Georgetown University Medical Sebd, Wasaisy- ron, D. C. HUR/.F.Y I F.F MOTLP.Y, M.D. Pro/ts- .nr of Medicine and Dlrreror, Cardb- Rryir.rory LoAarorory. Uwiversilr of Southern California Scttool of Medi- eine, I os Angeles. EDMOND ANTHONY MURPHY, M.D., Sc.D., Associate Pro/rasor of Rionarisrks .wd Mrdicbsr, The /oMs /loains University Seilool of Medi- cine. Baltimore. WILLIAM 3. MURRAY. Se.D.. Senior Srae Scknelsr, T4 )srctao. La4or.- 1ory, Bar Harbor, Me. RICHARD L. NAEYE, M.D_ Pro/euar and CAairman. Depr tnsrwt of korAof- olry. Pennsylvania SINe University Colr kRe of Medicine. Hershey. AI BFRT /1. NIDFN, M D., Pro/rswr of Mrdiawr, Orew PoMipaduNe Medical School aed University of Sourlserw ('alifornia; CAk/. Pafn.owry Disr.r Srcriow. Maniw Lwt,ee Kirre Hoqiral, t os Angeles. OAK RIDGE NAT1ONAl. LABORA- IORY, Oak Ridge, Teww. DONAI D M. PACf', PM D.- Pro/rssor a/ Physioloty and Director. lnuNrrr Z CrMr/ar RrsrrcA, University of rada, Lincoln. 81 ALBFRT B. PALMFR, Pit D., Auisran. lr../ns.w of lorhu(ugy, ()nircrsqr ul 7olcdo, Idcdo, (). ROSE MARIF PAN(:BORN, M S, Ar rutant F.aad Trchnoh.Rrv and Lrurrrr 1)r/.arrmrne of F.awf So ancr and 1 n A n.dosy, University of Ca6lornia,l)avn IOHN W. PARKER. M.D, Au..rar. lro/rswr of Parholo[y. University ol Southern Califo.nia ScMrol of Mcdi cone- I os Angeles. MARY SIFARNS PARS111 FY, Pnl) Auiu.nr lr../rss..r of Anatomy in c)S Mfries snd (irnr.aL.el. (olumbo. linirersily ('dkge of Physicuns ti %uo teons, New York. EDWARD W PEI IKAN, M.D., Cha4 nwn, Drr.rrmrns of PArmac..l..t) ond EsPerimrmaf Thrraoerrks, Boslnn Universily School of Medicine, sosron MAl.CO1.M C. PIKE, Pw D., Pr..ft.so. of Comn.rnny Mrdirinr ond Prdw rics, University of Southern California School of Medieine, Lo. Angeles. OTAKAR 1. POI.I.AK, M.D. Pw D F..errrivr Dirrctor, Dover Medical Re search Cenlet, Inc., Dover, Del. MORRIS POL1 ARD, Pw D., Dirrcra. 1riArnd L.Aorarory, University of Nolre Dame, No(re Danc, Ind. C. M. POMFRAT. PqD. Dirrrror of M"d.+~K.l RrsrrrA, Pasadena Fornda uon /or Medical Rnearch, Pwdena. Cal. S. N. PRADIIAN, M D. Prr D., Irofrr s.w of Phrmaroh.Rr. lloward Uni.er siry College of Medicine, Wasbingrow. D C. H. R. PRATT-TIIOMAS. M D- Pr..fn ar o/ l.rA.rlott. and llrsw, Med.c.l C.dlege u/ Sou11 ('.rulina, (Yarlcs/on PRO('FSS a INSIRI/MFNf CORPO RAIION, srooklyn, N. V. MARTIN S PROTlFI, flS, DDS, ('AaJ, Dr/vrn.rnr oJ Or.l ParAd..er. Newart (-ay HospiUl, Newart, N I WAI ifiR REDIS('H. M 1). Asr.Nl.re Pr../ruor o/ ('finrraf MrNriwr, New York University School of Medicine, and NY1/ Research Service, Ooldwaur Meanwial 11060161. New York.

I. Wlll IAM REOFI SON. M.D. rrofessor and CA.lrmaw. Drprtmrm of Mrd.ral Owcobty• Medical College of Virtinia, Rkbmond. HOBART A RFIMANN, M.D., rrofrs- sor of Medicine. Hasnemann Medical College awd Hospital, PWadelPttia. ROl1.AND C. REYNOLDS, M.D, Ar sisr.au rrofr»or of Pathology. Uwieer- si1~ of Tc.as Souw\wesrerw Madkal Scbol. Ddlas. ViCTOR RICHARDS. M.D. Chk/ of SrtM, PeesArleria. Medkal Cenler. Saw Frawcisco. SYILLIS H. RIESEN. Pti1.D, Srw/er /io- cfiewdsr, LJfe Sciences DJr/s1on, IIT Research Inritwe, Chicyo. R. H. RIODON, M.D. rrofrs.or of ra- (Adot~, Uwi.ee.My ef Tear Medical fManck Odeessow. SYDNEY C. RITTENBERO. P+M D., rro/rs,oy of glarreriolap. Uwiversilr of So.(hee. C.Iifosra. Le. Anteles. IIENSON 1!. ROE, M.D., Assoriarr rro- lrssor of Srt"; CMrf, Car/iar Sr- (e.y. Uni.er..y of California Scboi of Medieiwe, Sa. Francisco. WAYNE 1.. RYAN. Pw D.. Prnfessor of fl.nclvmirrrr. Universitf of Nebuka ('ollqe of Medicine. Omaha. PETER F. SAI.ISBURY, M D., PH D., Head. Inttnsire Trratmrnr Center. Saint )osepb Hospital. Burbank. Cal. . PAUL SALTMAN• PH D., .Nsinanr rro- /essor, Dqarrmrwr of Siochrmimy aw/ Nrrrinow. University of Southern California Schod of Medicine. Los Antcks. ULRICH H. SCHAEPPI, M D., Dirrc- tor o/ Nero~Arm.roloty• Mason Re- searc\ les(Nwe, Worce.ler, Mw. IOROEN U. SCIILEGFI., M D.. Pu.D, rro%ssor an/ CMirweaw. Drprtwuwl of Srtery, Tulawe Universily Sc6ool of Medicine. New Orleans. AI.VIN R. SCHMIDT. Pw.D., Director of CownxNwg, T.f1s Universiy, Med- ford, Mar. ISAAC SCHOUR. D O S.. tM.D., DSc, Dean. Uni.ersi(y of Illinois College of Dentistry. Chicago. MAURICE S. SP.GAI., M.D., Cl/nicd rrofrssor of Alydkinr, lulls Univer- sity Schod b( Medkine; Director. De- rertmewt of Inhdatiow Therapy. floa- ton CNy Ilo.Pilal. Boslon. LUCIO SEVF.RI, M.D., Director and Draw. IwsNrrre of Anatomy and rarAd- U y. Diviaiow of Canoer Researcf4, .i.ersilf of Pervtia, Per•.{iR Italy. CHARLES 1'. SHERWOOD. M.D. As- sistawr Professor oJ Ra/lologP. Uni.er- sNf of Rochester Sc6ool of Medicir and Dentistry. Roclrsler, N. Y. SIIO11 SHIBATA, M.D.. PN.O.. Profrs- .or of rharwwcototr. Univeni(y of Hawaii School o/ Medicine. /lonohdr. /OSEPH H. ROOERS, M.D. Holl N.rne ./ lesw Hospital. O.d.de.< Ala. ROBERT C. ROSAN. M.D.. AssocLN nofewr of raeAoloty and red/Nrlcs, 31. Lwk Uni.crsily School of Medl- siws• Assocfae rarAelotisr. Cardinal Ok:ne. Ma.orW Ho.tAd far CW- 4eq St. Lo.k. JOHN R. ROWLANDS, Pf(.D.. Stas SckrW, Sr>Mlewesl Researcb Lrilule. Saw Aalowio4 Tea. .11P.NIAMIN A. Rl1BIN, PIS.D Assistant rrofrssor ./ rrF/k Ifrdrk, Ba~{or Uwi.enil~ Cotaefie of Medicine. Ilo.- sle.. RONALD P. RUBIN. Pw.D., Professor / r/+orw.acototy, Medical College of vYtinia. Ricbnowd. HPNRY 1. RI/SSPK• M D Prrrllrnr, The Ru..et Forndalion, ine. Staten Island. N. Y. W. C. RUSSPI L, M D, University of Te1a. Medical Center. Houstow. DAVID L. SIMON. M.D.. Ins(rrtor lw Internal Mrdkiwr, Cincinnati (knerJ Hospi(al, CiwcinnNi. PRIK SKINHDI, M.D., Chief. Deprr- mrwr of Neurology. BisPeefer. Hoqi- td. CoPenhaten. THFODORE A. SIOTKIN PN.D. As- siss.nr Profruor of r"On.colot). t)u\e University Mcdical Cenler, Dur- ham. N.C. 82 I (IEOR(;E W. SMETTFRS, M-D., Auo- cietr in ratAolotl, Northwestern Uni- versity Medical School, CCticqo. (IF.NF. M. SMITH. h(.D.. Assistant rro- /rrwr of rsy.holoty, Harvard Medical School. Massachusetts Oeneral HosPi. ul. Bos(on. LUCILE SMITH, PN.D., rrofrssor of sarchrmisrry. Dartnwulh Medical School. Hanovcr, N. H. SHELDON C. SOMMERS. M.D_ Dbee- sor of LAorarorks, Lenoa Hill Hoe- pital; Clinira/ rro/es,or of Pathology. Colurnhia UniversM~ Collqe of Pwysi- ci.ns i Surteonk New YorY. FRNEST SONDIIP.IMlR, PN.D, Aaso- ciatr rrofr»or of Aiorhetnlsnl. Cd- kge of Forestry. StMe Uwiversi(y of New York. Syracuse. T. M. SONNEBORN. Pw.D, D4rM- f rrshd Service r rasor of Zoelop, wdiawa University. SAM SOROF, PND., Rra/, Drprswsrw of Macro/nolfcYir CAewris/ry, The Institute for Cancer Researcls. Phila- delolia. SOUTHWEST RESEARCH INSTI- TUTE, Saw Anlo.iq Tea. DAVID M. SPAIN. M.D. Di.r~esor~ Dr- ~..fine/V o1 PNhofot), TII! ~rOOldak Hos0i1a1 Center. Sroollm N. Y. AI.F.XANDER SPOCK, M.D., Asslstrw Professor of lediadlYs, Dake Uwl.er- sar. MMedical Ce.kr, Drrha.r, N. C. FREDERICK /. STARE. M.D Prefrs- sor a/ Nrtrls/ow, Har.ard JnlrersNr School of Prblk lkaRll. Boston. C. HAROI D STEFFP.E, M.D_ Director of LaAorauorks, Mdhodisl Hoyild, Memphis. JACK P. STRONO. M.O, Assor/ue Pr.•fruor of Pathology. Lorrisiawa Sta/e Ilnivcrsiry School of Medicine. New (kkans. MARION B. SULZBPROER, M.D.. rro• leswr and Chairman of DrrrwuoiotP and Sy~Ailoloty, New York Unive.- sity Belk.ue Medica) Center. New Yort. RFNATO TAOIURI, hr D. Associate Professor of Psychology. Oraduate School of Business Adminis(rtlion, Harvard University. Boston. CAROI.INE NEDFI.1. THOMAS, M D-, Profnvo I.mrruwr of Afrditlnr, lie Johns Ilopkins University School of Medicine. Baltimore. JEROME F. THOMAS. PH D-, rrofntor of Sanitary i-ntinrrrint, University of California. Berkeley. JAMES E. P. TOMAN, 1M.D., rrofrs- sur and Cha/rwraw. I)rparlmrnt o f PIA.r - macofcyfy. Chieato Medical School. In- slilute for Medical Research. Chkajo. JANET TRAVEI.L• M D• Associate Professor of CUnkal rAarmacolotr. CorneR Universily Medical Cdkge, New Yoe\. LIE SHA TSAI• PN D., Research Asa.- ciate iw rarholotf. Yak University School of Medicine. New Haven. Cown UNIVF.RSITY OF SOUTHERN CALI- FORNIA. l.o. Anoteks. EI.I.IOTS. VF.SELI., M.D, rrofe»o.awl CAabwraw. Drpartmewr of PArns.rol- oty, Pennsylvania Stue Unnersir/ Cd- kge of Medicine. Milton S. Hershey Medical Cen/er, HersAey. ROMEO A. VII)nNP., M D., Associatr Professor of Pathology. Yak Uni.er- sitr School of Medkine, New Ha.en, Conn. PETER K. VOOT, Pw D., Professor of MicroAioloRf. University of Wast.int- (on School of Medicine. Seattle. E. D. WARNER. M D., Professor of ra- rholot' State Universutr of lowa Cd kte of Mcdkine, lowa Ci(y. SHIFLDS WARREN. M D., D/rrctor of I.eborarorire• Cancer Research /wstl- srrr• New England Deaconess IIo+Pi (a1. boaon • YASUS/II WATANABF, Pnl), Asu. .ratr Mr.nArr• The Wisur Institute of Anatomy and Biology. Philadelphia. BARBARA K WATSON. Prr 1) • Anlu- awr Awrrrri.doRdrt, Ma.saciwseNs (len eeal I/ospilal; Rrrrar.h Mu.rlatr, tn sa.trri.d..Ry .wI Immrnainrr, /l.r vard Medical School, Boa.w. lOIIN S WAU011, Pn D. Professor of CAreunuy, Massachuseus Institute of lechnnloRY. Camhridte. aI

I RICHARD 1.. WFCIISI.FR. M D• Cliw- kal rhyrA.lncirr, Momelkwe Hospital Instirwe of Researcb• riNlslwrgh. JOHN V. WFII.• M D.• Aisirranr rro- leswe of Mrdkine• University of Colo- r.do Medical Cemer, Denver. A. WEINSTOCK. Pw.D, Research t/o- ~ chearJ.r, Li/e St/encer Di.idoR, IIT Reavch insriaute. Chicyo. RUSSELL W. WF.I LER, M.D_ P.rhol- oRFir• Merwwial Horqiral of Chester CowNy. Wea Chesrer, Pa. A. STANLEY WFLTMAN. Pw D., Aaso• el.re Pro/isser of rharwroroloty and Re.raech, BrooLly C.Rqs .1 Nv- teacy, Ilroollfw, N. Y. SIMON H. WENDER. Pt.D_ Rtar.r<t Pro/euer of lliocMwr6ory. U.i.ersily e/ bklaborn.• Noen..w. DUANE 0. WENZEL, Pu.D., Prolesrom and Ch.ha..w. Depara.ent of Phwnw- tvlo4y and ro.kolaly, T4e University of li.s.s Scbooi of lif.rrn.cy, I.aw- renoe. FRFDFRICK F. WIIISKIN. M.D. C.M.. Dirertor, Dirisien of Health and rer• sonality F.aailiheium• The Age Center of New England. Inc, Ikrslon. ROOFR l. WILLIAMS• M.D-, Professor of Chr.airrr'; Director. Clayton Fore- derioe IfincAemkal lwnUidr, The Uni- versiq of Teaas, Arstin. DANIFL H. WISF.MAN• M D., Aasiu- anr Pro esi.r of hdutrkr• Universil~ of Sow w Cdifornia; ('hildrew's Di- •isiuq Los Angeks Counry (leneral Hospital• Los Aqeks. 1. EDWIN WOOD. M D.• in.nuctoe In hledkusr, Bosrow Uni.ersUy Sehool of Medicine• Boslow. SUMNER WOOD, )a., M.D_ Asabrawr Professor of r.thdog ~~ Tle loMr HoN<ins U.i.ersiry Sc6od of Medi- cine• Bdtiwwore. JOHN P. WYATT, M D., rro e.uor o/ Pathology. Saint 1.ows niversily School of Medicine. St. Lari.. I i I CNnEX OF PRINCIPAI. INVESTICATORS Ahood• L. O., 54, SS Arcos. 1. C., 14, 15 Aviado, D. M., 2• Bhagal• B.• 56 Bing. R. 1.. 41, 42, 43 B(.kn, (i., 57 0 IlossE. R., 64, 63 Castro. A.. 61 Chalon. 1.. 16. 29. 30 Cochrane. C. (3.. 46 Cohen. A. B., )1 ('osla• P. T.. lr., 64 (-ross. C. E., 31, )2, )3 Essrnan, W. B., 57 Fishman. W. H.. 16.17.18.19. 20 Friedman. (i. D., 65 Fursr, A.. 28 (;oldstcin, L.• St Hamosh. P.. 34 loachim• H. L.. 19 Kixtri• R. t:., 25 (.auweryns, J. M.. 35• 36 I cvy. 1. A.. 21 ManhoW. 1. H., Jr., 21 Mason, R. Ci., 31 . McKennis, H.. Jr.. 59 Meier, H. 22. 2)• 24• 6) Repn• T. J.. 52. 53 Riftin, D. B., 26 Ruse. C. 1... 65 Ryan. U. S.. 36. 47. 411, 49. S0• 61, 6. Saslry, B. V. R• S9• 60 Sloane. N 11., 24 SobA. 1.. A.. 44. 45 Saxnmers• S. C.. 27 lravis• J.. 37• 3S• 39 Vatner, S. F.. 52 Wernhaum, (3.• )9, 40 Wenzel• D. (;., 46 y 84 es

INI)EX OF SENIOR AUTI1l)RS Abood. L. G-, 55 Ahmed, S. S • 53 Arcos. 1. C.. 13 Arlus• M. F.• 14.15 Baugh. R. 1., 37 Bedigian. 11. 0.. 22. 23 Bhagat, B.• 56 B1ng. R. 1.• 41. 43 Boden. cl.• 57 fossl, R.• 65 Caqro, A., 61 Chalon, 1., 16, 30 Chang. C•H., 17.18.19 Chiu. A. T., 49, 50 CocMane. C. O.. 46 Cohen. A. B.. 31 Cosu. P. T.. Ir., 64 Cron, C. E.. 32 David, 1. S. K., 44 Di.w+. B. A.. 24 Doyk, 1. L.. 21 Es.+m.n, W. B., 57 FisM+an• W. H.• 16. 20 Fos, R. R.. 22 Fricdman. 0. D..63 Frkdman-Mor, Z• 29 Furst A.. 2! Hamwk M.. 34 Harbi.on, R. D.. 59 Hus..is, M. Z.. 31 lo.chirw, H. L. 19 Ito• H.. 28 lohnson• D. A.. 37. 39 Katz. 1. S.. 30 Kimbel, P.. 40 Kouri. R. E.. 23 1 auweryns• 1. M.. 3S• 36 Levy, 1. A.. 21 I.owy. K.. 54 Moschos, C. B., 52 Nelsen, 1. M., St Pcirce• T. H.. 32 Rlfkin, D. D., 26 Rose• C. L. 64 Ryan. J. W., 47. 4e• 30, 61 Ryan. U. S., 36.48. 62 Saba. S. R., 51 Sarma. 1. S. M., 41. 42 !43 Sastry. B. V. R.. 60 Sloane. N. H., 24 SoioA. L. A.. 44 Sommers• S. C.. 27 laveira da Silva. A. M.. 34 /ravil, 1. 39 Varma• K G.• 45 Valner• S. F.• 52 Weinhaum• (7.. 39 Weisbroah• S. H.. 61 Wcnz.cl, D. O., 46 Wilsoa, K. L.. Jr.. 59 York. O. K., 33 I 4M