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SCIENTIFIC ADVISORY BOARD to The Council for Tobacco Research-U.S.A., Inc. as of December 31, 197R SHELDON C. SOMMERS, M.D., Chalrtnms Direcw of [.aboratorles. Ltoos HiU Hospital Clinkal Professor of Pathobp Colk~e of Physkiana fc Sutgeorr o( Columbia University New York, New York RICHARD ). BINO, M.D. Director of Cardiology and lnt. snard MrdkJne Huntington Memorial Hoapitd, Pasadena, California Pro/essor of Medicine University of Southern California School of Medicine Los Angele., California JOSEPH D. FELDMAN. M.D. Read, Department of Immunopathology Scripp~ Ctink and Research Foundatbn La )olla, California WILLIAM U. OARDNf'R. Pw D. Sclenti/ic Director. The Council for Tobacco Research-tI.S.A , Inc. E. K. Nunt Pro/essor of Anau„ny (rnuritur) Yak University Schocd of Medicioe New Haven, Connecticut ROBERT J. HUEBNER, M.D. Chief. Laboratory of RNA Tumor Viruses National Cancer Institute Betheada, Maryland LEON O. IACOBSON, M.D. Joseph Regenste(n Pro(rstor of Biolotkal Scirnces Unive of Chicaso Chkago, Wnoia HENRY T. LYNCH. M.D. Professor and Chairman I Department of Preventive Medicine and Public Health 0eighton llniveraty School of Medicine Onaha, Nrt'rarka t HANS MEIER, D.V.M., Dr. Med. Vet., M.R.S.II. Senior Staff Scientist The Jackson Laboratory Bar Harbor, Maine LEE W. WATTENBERG, M.D. Professor of Pathology Department of Laboratory Medicine and Pathology Umversity of Minnesota Medical School Minneapoli., Minnesota JOHN P. WYATT, M.D. Director Tobacco and Health Research lnatilute University of Kentucky Lexington. Kentucky Sdenliie Staff of 7Ue Ceuneil WILLIAM U. GARDNER, Pw.D. SdenH/ic Director ROBERT C. HOCKE7T, PH.D. Resewrh Director DONALD tl. FORD. P41.D. VINCENT F. LISANT7, D.M.D. Associate Research Director Astoclate Research Director DAVID STONE. PH.D. Associate Research Director ,

CONTENTS Introduction . . . . . . . . . . . . . . . . Abstracts o[ Reports . Cancer-Related Studies . . . . . . . . . . . The Respiratory System . . . . . . . . . . 1leart and Circulation . . . . . . . . . . . Neurophannacolo6y and Physioloq . . . . . . . Pharmaeolosy and Biochemistry . . . . . . . . Immunology and Adaptive Mechanisma . . . . . Epidemiology . . . . . . . . . . . . . . Active Projects . Completed Projects . . . . . . . . . . . . . . lode: of Principal Investisaton . . . . . . . . . . lodes of Seoior Authors . . . . . . . . . . . . . . 5 . 7 . . 7 . . 26 • • 41 . . 49 . . 62 . . 70 . . 77 . . 83 . . 92 . . 103 . . 104 I ' Introduction This report m.riu the completion of 25 years is which 7b Council for Tobacco Research has supported a program that has bewrne sb" world's mosl extensive noa-governmeatal research effort rekvaet Io smokins and health. When The Council was estaMished in 1931, it adopted a basic policy that has been followed without deviation: to pl.ce responsibility for research policy and programming in the hands of a Scientific Advisory Bo.rd and to have 1ho research conducted by independent investiplon in tht3r own lnstitutions with so striop attached. Up to the end of 1978, the program has resulled in the publication of 1.622 reports and articles that acknowledged Council support. 77rua, the pro- gram has produced considerable data related /o smoking and health during a periud Ibat also saw Ihe generation of oomiderable eoMroveny and unotior about the subpct. No one can fully predict at this time what these publications will meau in the search for the answers b the problem of the major, a8ing-.»ocialed and constitutional diseases of eancer, heart disease and chronic pu4aoa.ry ailments. However, the hope is that some day the Andinp from one or mrore studies may provide keys or links for a major biomedieal advance. The Council eaisu today because the complex etiology of these oonslitu- tional diseases remains unraveled. These diseases have been associated ststiL- tically with amoking, but such a»ociations are not proof of cause and effect. The diseases are extremely complex-cancer itself, for eaampk, b siol os disease but many-and /indin8lheir causes will not be easy. The overall importance of genetic faelors is being increasingly reco8nired, accepted and studied. Progress is being rwade, but it is necessarily slow sad painstaking. The statement of many years ago that we live in a "sea of careLa{ear' is supported by nmch recent and current research. Many Ihin8l--both natural and man-made-that we ea1, drink and breathe have been implicated (n canoer and heart disease, at least in laboratory eaperimenls. But exact proof, proof that meets scientific criteria, is often lacking. Whatever contribution lo scientific knowledge that may be attributed to The Council's program, as evidenced in part by the numerous reports in the Gterature, is due principally to the members of the Scientific Advisory Board, p.st and present. 7lrese scienliMs, who have always maintained their inuilo- Iiond alfiliations, have given unsdOshty of their time and talents. 'i'he result of their combined cRort lor so many years is an Imaginalive undertaking that has enabled many reaearchen to pursue ideas and theories that might other- wise have gone une.p{ored for lack of support. lbe Council is gratified at the response through the years of Ihe sckn- tists who have applied to it for research support. 'Ibis intereat remained high in 1978 aand the number of requesu for support of new and continuing re- search assures a worthy future. 7 his report contains abstracts of research findings published in 197• that aCkMtwtrdeted ('ouncd sponsorship. As can be aeen. Ihe major em~hasis c.xr- linucs to be directed loward cancer, the cardiova+colar system and Ihe respira- 3

tory tract. The abstracts of the published papers are indicative of the areas of research supported by llre Council. lhe Council looka back with utisfaclion at what has been accomplished in the lasl quarter-century. It looks to the future with optimism and hope for research that will help sdve the mysleria of canter, heart disease and chronic pulmonary ailments. The Council pkdgea to continue its support of inde- pendent investigators in their effotts to add to scieotific knowledge related to smoking and health. I 1. ('.awcer-Relwted Studies Abstracts of Reports Following re ab.trac/s, approved by the aulhors, of reporu oo .ew research acknowkdging wpport front TLe Council that have appeared in acka- tifk journals since publication of the 1977 Report. The name of Ihe recipieM is in iulica. The abstracts are grouped under these headings: 1. Cancer-Related Studia, (1. lhe Respiratory System, 111. Neart and Circulation, IV. Neuropharmacubpr and Physioloty, V. Pharmacology and Biochemistry, VI. Immunobp aad Adaptive Mechanwns, VII. Epidemiology. CRITERIA FOR Se1.ECi1NO CHEMICAL COMPOUNDS FOR CARCINOGENICITY TFSfINO: AN ESSAY The ever increasing number of organic compounds conslantly being bstra duced into the various aegmenls of the wrorldwide economy cnrales 1he aeed b devise a formal scheme of selection criteria for testing of suspectod car- cinogens, and to establish priorities /or auch testinll. Four caleitoriea of erileria must be considered: (1) structural. (2) operation.l, (3) "guilt by arocia- tiwr," and (4) "after the fact." This essay Rives an analytic discussion of each of these criteria. Structurd criteria are deduced from a=eneral perspective of the structure-activity relalionship of known earcitsoRens. The operalional erih teria, which are complementary lo the structural ones, represent the sum of the bio-/unctional capabilities of chemical compounds. These capabilities oor- relate with the ability to induce malignancy lhroukh such mcchanisma as, for eaampk, mulajenicily, induction of DNA repair, and Mnmuno.uppression. The "guilt by association" criterion alates that a compound belonging to a chem- ical clase already known to contain several other compounds eslablished as potent and mullilarilet carcinogens should be 1esled under more stringent con- ditions than those of the standard bioasay. The "atter the fact" criterion deAnes previous epidemiologie indications as the basis lor selecting certain chemical agents /or carcinogenicity tesling. The overall significance of Ihe bioassay dala obtained on individu.l eompounds, however, should be ea+- sidered in conjunction wilh other endogenous ud eaoaenous interacting lac- tors such as: /1) synergistic and anlakonistic sKtivoy between multiple car- einokenie agents; (2) dlects of nonc.rcinotenk environmental agents on carcinogenic activity; and (3) viru.es, nusrition, radiation, age, stress, endo- erine bdanee and th, state of th. (rnnwne wrveillance system. 'fln huae number of akenu and facton, their interactions and the coesidence of fre- quently conflicting societal soala, indicate the seed for the establishment of versatile acientificoletal standards and guidelines applicable to eatetoria sarl types of chemical agents rather Ihan to individual eompounda. ~ /rror, l. C. ~ lorrnd of Ewrironm.nrd laholosy ad Toakoloty 1:1))-ISe, 1978. 7' 6

OtRer aupportr National Cancer Imtilute. From the Seamen's Memorial Research Iaboratory, U. S. Public )ler,lth Service Ilospilal, New Orkana; and the Department of Mcdicioe. Tular c Medical Center. New Orleans. UI:1 RASTRU(`Tl1RAL AND METABOLIC DETERMINANTS OP RINISI'ANCE TO AZO-DYE AND SUSCEPTIBILITY TO NI1 ROSAMINE CARCIN(7GENPSIS OF THE GUINEA-PIO The a:o dye rcductase and nilroaamine dealkylaae activities of normal guinea-pigs and rala were compared b those of animals fed ar.o dye and niuo.amine. Electron microscopy reveakd ultraalruetur.l alkralions of the guinea pig liver during nilrosamine administration. Dielhylnitrosamine (1)EN)- induced hepatic lumot cells showed extensive proliferalioo of the rough endo- plasmic reticulum ( ER ), while the smooth ER wat quite tp.rx; the opposite was true in the prema)ignanl liver. In the rnt, however, administration of either DEN or Y-methy/-Idimelhylaminoa:obtnzene (Y-Me-DAB) caused prolderatioe of the smooth ER and sparsity of the tough ER in both pre- malrgnanl and malignant tissue. In both specin, the number of ribosomes on the outer surface of Ihe liver rough PR was greatly reduced as was the RNA/ protein ralio, which correlated with a decreased r+eaponse to an Slt probe In nmicroaomal suspensions A:o dye reduclase activity was higher in untreated rats than in untreated guinea pigs. After sit weeks of T-Me-DAB feeding, however, this was 76% lower in the ral while there was no significant decrease in the guinea pig, which is refractory to azo dye carcinotenesis. 77sus, as in- geslinn approached the lumorigenic threshold doae. the rat liver's ability to inactivale the dye was much impaired, but not that of the guinea pig. Control levels of ni/rosamine dealkylase were idenlical in both apeeies, however, and remained essentially unchanged Iter 10 weeks of DEN adminislration. Since nitro.amine dealkylation represents activating metabolism. this provides the basis for the idenlical susceptrbililies of the rat and the guinea pig to DEN carcinotte.rcsis. Of the two enzymes eaamined, only Ihe guinea pig azo dye reduclase appean to be independent of glucose repreaion, since its activity was unchanged by starvation. Statvalion-irduoed elevation of azo dye reduclne activity in the rat was not affected by Y-Me-DAB administration and only slightly influenced by DEN In both specks, however. DEN abolished the aarvation-induced increase of nilro.amine dealkylation, while )'-Me-DAB decreased it only slightly. Bryant. (i. M., Sohal, R. S., Arpn, M. P., and Arcos. l. C. drittrA Joucwd of Cancer 76:676-691, 1977. Other .u pport ) National Cancer Institute. Prom the Department of Mcdicine, Tulane Medical Center and Seamen: Mernorr.J Kcseaah I aN.ua1ry, 11 S Public Ilcalth Service llolpilal, New Ur1c..ns,..uJ Ihc Ikp.rrotcnt of Itwhity, SanUhcrn Melhodnl llurverslty, 1).IL1. I IiYDROCARBOMNrI'ROSAMINE PULMONARY SYNCARCINO- GEN['SIS: RI:CIPROCAL EFFECTS ON METABOLISM This study reports on the entynqlogical correlates of the pulmonary ayn- carcinogenesis which h.a been observed between melhykholanthreoe (MC) and dimethylnitrosamine (DMN) following acute administration of the car- cinotenr. E.perimenlally, .ryl hydrocarbon hydroaylase (AHH), eposide hydrase. DMNdemelhylase, and ilIvlalhione-S<potide Irana/eraae activities were studied in the liver and lung of DBA/21 and C378V61 mice. two atraiaa which are, respeclively, noninducibk and inducible for hepatic A1111. Change in theae enzyme activities was determined following acute adminislralioa of MC. DMN, and their combinations. In the liver-eacepl tor a.ubstantial ie- duction of hepatic Atilt in the C37B1./61 alrain-1he metabolic pattern is very similar in the two strains. In both slrairr DMNdemelhylase ia highly sen.utive to DMN prelrcatment at the aia mg/tti level. Increase of DMN to the total syncarcinogenic dose of 60 ms/kil abolishes induced AIIH in the C37B1./61 and brings about substantial inhibition of aB other en:ymea studied. In the lung of the two alrains, induced Atilt was found to be highly aensitive to DMN pretreatment in the DBA/2J, and DMN-demelhylaae 11 (the only DMN-demethylse present in the lung) was substantially reduced following Prelrealment with 60 mg but not with 6 mg DMN. Tlse most interesting re- sulls in the lung are .een with the C37BL/61 strain since, with 60 mg DMN/ tg, there is a substantial decrease of the epoaide hydrase together with a substantial increase of the AIIN. This suggests a mechaniatic interpretatiom of the Atilt increase within the framework of the known palhways of hydro- carbon metabolism and a greater suaceplibilNy of the C378t./61 strain to Ur combined administration of MC and DMN. Arcoi, J. C. d at. In: Gelboin, II. V. and T: o, P. O. P. (eds. ): rotpcycflc Hrdrocarbowr md Cancer--Environnrent, Chemistry and Alttaboll.ns, New York: Academic Press, 1979. vol. 1, pp. 271-282. From Tulane University School of Medicine, New Orkana. DIPFP.RENTIAI. EFFECTS OF a-NAPTHOFLAVONE AND PREGNENOLONE-16a1:ARBONITRILE ON DIMETIIYI.NITItO- SAMINN-INI)ll('I:D IIEPATOCARCINOGENFSIS 7he carcinogenicity of dimethylnilroaamine (DMN) is inhibited by the co-administratiun of either 7-melbykholanthrene or anminoacelonitrik, com- pounds which substantially lower the activity of DMN-demcthyla/e, a micro- somal mi.ed-lunction oaidase that catalyzes the activation of DMN. In the present investigation imed at elucidating further this correlalion, the eRect of administering jil-naphthoflavone (0-NF) and pregnenolone-16.-carbonitrile (PCN) on the hepatocarcinotenicity of DMN in male SD rata wa%* explored. Bo1h /! NI: and 1'('N are potent represson of the bw-Michaeliu-conslant en- :ymalic form of 1)MN demethylase. Ncpalie tumors were found in every group of rats receiving DMN alone or in combination with A NP or PCN. 8 9

I I I a The morphology of these tumors waa quite similar for all three kroups, with 3396 of the 53 liver tumors observed being angiosarcomu. DMN-induced bepalocarcirw6enenis was partially inhibited by PCN and enhanced by P-NP. Seven liver turnors were found in 45 rats fed DMN plus PCN compared to 32 liver tumon in 43 rats fed DMN plus aNF; 14 liver lumon wrre found in 43 rats fed DMN abne. No liver tumon were detected in rats that received only PCN, /f-NF, or Ifse administration reAicks. The unespected enhancement of DMN Induced carcinokenesis by aNP prompts a reexaminatwrn of the presently accepted activation mechanism of DMN. Argu., M. F., Hoch-l.igeli, C., Ars•or, !. C., and Conney, A. H. /orrnal o/ the Narional Cancer Iwsrhr/e 61(2) :441-449, 1978. O1Mr arpportlt National Cancer Institute. From Ihe Seamen's Memorial Research Iaboratory, 11. S. Public Hcalth Service llospital, and the Department of Mediclne, Tulane University School of Medicine. New Orkans. USE OF HInH CONCENTRATIONS OP DIMETHYLNI71tOSAMINE IN SACI-I!RIAI. LEl HAI-1 fY, MUTAOENpSIS. AND flNZYMOL(X)ICAL STUI)I PS Previous studies have established that the potency of dimethylnilro.amine and dielhylnitrosamine in bringing about conformational changes in protein is comparable to that of urea and kuanidinium chbride, well-known protein- denaturing agents. In addition, an optical rotatory dispersion study of the ovaR>tunin conformalional change as a function of dimelhylni/rosamine coo- centration had indicated a concentrationdependent ambivalent effect on pro- tein conformation. Thus, at concentrations of, and greater than, about 200 mM, dimelbylnitrosamine brinp about denaturation. However, a1 concentrr tioro from 200 mM downward the b, constant of ovalbumin increases with the decrease of dimethylnitrosamine concenlratioe, suggesting Ihat, in this oon- eeotratioa ne6e, dimelhylnilroaamine bas a refolding ('Yightenink`) rather thae unfolding (denaturing) effect on the protein eecondary structure. 8ecause of ita protein-denaturing ability, dimethylnitrownine at high concentrations can bring about cell death. In eonlrast, when 20 to 200 mM dimethylnitrosamine is used in esayme and mutagenesis awys, because of its re(olding cffect on protein secondary slructure at thne ooecentrNions, il may produce a1Mtileric enzyme eonformational changes iw vhro, thereby possibly creating artiiactual phenomena. Indeed, the use of nonpharmaeologically high eoncentrations of dinxthylnitroaamine and diethylnhrosamine appears to be the basis of con- flicting reports on the repressibility versus inducibility of dimethylnitrosamine demethylase and diethylnilrosamine declhylase and on the effect of enzyme indu.er prctre.lment on ndro.amine mulagenicily activation by microsnmes. keaenr onvesrrpu.a.s sugaeN rhst the apparent confbct is due to the eslsrence ot h.o rnlymK /orme o/ dinuthylmuosamine demerhylase that rc%porwl in duiuctn,.11r opt.•vu +ass (n.mely, repreasn+n and mdutthun) ts- cnryme in,tu.cr plctrratmrnt Il.r.r ot.xrvahuns and othcrs kad the aulhuu so con- 10 I clude that the cortfentration of dimethylnitrosamine (or any other nilrosamin.) uud in In vitro assays should be kept under 20 mM, and preferably below 5 mM. Since the purpose of studying the melabdism, mutagenicily, and cellu- lar effects of dimelhylnilrosamine and other nilroumina Is principally to pia insight into the mechanism of their carcinogenic eRecl, physiologically realistic testing levels should be observed. Argu., M. F. and Arcof, l. C. Cancer RriearcYt 3t:226-22t1, 1978. From the Seamen's Memorial Reseanch Lboratory, U. S. Public Health Servlo. Ho.pital, and the Department of Medicine, Tulasse Medical Center, New ('rkans. TISSUE AND SUBCELLULAR DISTRIBUTION OF a11-DIOXANC IN THE RAT AND APPARENT LACK OF MICROSOME.CATALYZPd) COVALENT BINDING IN THE TARGET TISSUE Diosane, a commonly used induqrial and laboratory aolvent, lua bees known to eahibil both bxic and carcinogenic activity. As part of a series of investigations aimed at elucidating the mechanism of these eRecN, this paper presents data on the distribution of 01l-dioaane among a number of rat tis- sues and various wbcellular (ractions of rat liver. At various times dter intraperitoneal injection. d'aaane was distributed more or less uniformly among varqus tissues (liver, kidney, spken, hrng, eobn, and skeletal musck), which is consistent with iu polar/nonpolar nature. llowever, in eoMraN so this nearly uniform distribution, uudies of the nature of binding revealed larp tisxre differences. Ttre liver (the main target of careinogenesi.), colon (de- acending segment), and apken showed a greater amount of "eovaknl" binding as measured by Ihe incorporalion of radioactivity Into lipid-free, acid-insoluble materials. Much kss "covalent" binding occurred in the skeletal muscle and blood. Investigations of the aubcellular distribution in Uver Indicated that moN of Ihe radioactivity was in Ihe ey/osol, followed by the microsonul, mito- chondriat, and nuckar /ractions. The binding of dio.ano lo the macromole- cuks in the cytosol was mainly noncovaknl. The percent covalent binding was highest in the nuclear (raclion, followed by milochondrial and mkrosornal fractions and the whole hotnossenale. Pretreatment of rals wilh inducen of mkrosomal mi.ed function olidasa had no aignifkant effect on Ihe covalent binding of diosane Io the various subcellular fractions of Ihe liver. 7lrere is no micrr>M>toetatalyred /w vitro binding of diosane to DNA under conditions in ..hich there is ulensive bindinrof benro(a)pyrene. Woo, Y-T., Arrus, M. F. and Arcos. 1. C. LI/e Sciences 21(10) :1447-1436, 1977. Olhtr support: National Cancer Inslitute. From the Seamen's Memorial Research I aboralory, I1. S. Public Ileaith Senkaa Ilosprul, and Ihe Department of Medi.cine, Tulane Univenily, New (bkaos II

STRUCTIIRAL IDP.NTIFICATION OF p-DIOXANE-2-ONI3 AS 719@ MAJOR URINARY Mt'TABOLITE OF pD1OXANE Intensive diosane eaposure causes severe liver and kidoey daroage and even death. The compound is also a hepatie carcinogen in rata. In 1.lis inves- ligation of the In viro metabolism of diosane, gas chromatography ((iC) of the volatile compounds present In the urine of rats administered dumane re- vesled a maior metaholile. The latter was detectable only at low 1 N kveA, and the amount escreted was bo1A dose- aod /ime-dependent, reachin,: a maai- mum 20 to 28 hours after intraperiloseal intectioo of dioxsne. Administration of diethykne glycol produced the sarns metabdite, but mostly withir, the Brst 16 houn, suggesting that this compound may be an intermediate in the iw viro metaholism of dioaane. Under similar eorditions, ethylene g!ycol, di- glycolic acid and osalic acid did not induce escrelion of the metabolite. The isolated and purifkd melslwlite displayed an intense carbonyl band at 1,750 em-' In the Infrared (IR) spectrum. Its nuclear marnctic resonance (NMR) spectrum indicated two Iripkts and one singlet of equal Intensity at 1.81. 4.42 and 1)7, respectively. Upon GC-mass speclropholomelrie sludiq, there was a parent peak at m/e 102. The nsetabolite wY idenlified u p-0toRSne-2-ooe, as conflrmed by comparison to ita synthetie reference eompound, .rhich ea- hibiled the same IR. NMR, and GC-masa spectra as the unknown. A tenla- live metabolic pathway for diosane is presented; micraaornal mixed function oaidases appear to be involved in this pathway. Preliminary eaperim:nl/ indi- eate that the melaholite is considerably more toxic than the parent compound. Its potential health hazard remains to be asaessed, espeeially since it is used as a commercial and industrial preservative. Woo, Y-T, Arcoi, l C'., Argus. M. F.. Gri/fin, 0 W., and Nishiyama, K. Nounyn :jchmlcdtbrrt i Arrhivct o/ P.rrAoloeY 299.2N )-287, 1977. Other supportt National ('ancer Institute From the Seamen's Memorial Research I ahnrstory, I/ S Public Ilealth Service Ilospilal; the Department of Medicine, Tulsne Universily; and the fkpartment of Chemis/ry. University of New Orkans, New Orleans. EFFI?fT OF MIXED-Fl1N(TION OXIDASE MOI)1f7F.RS ON MNTABO1 ISM AND IOXICI CY U1' 1118 ONC(Xil?N 1)IOXANH The effect of inducers and inhibilon of mised functie.n oxidases (MFO's) on Ihe in riro rnetabolism of dio.anc in the rat was sludicd with both non- radioactive and "('-labekd dioxane. In addition, the effects of various inducen on the acute tosicily of diosane and Its principal urinary molaholite, p diosane- 2-one, were eaamined in an attempt to elucidate the relationship between the mr1alm.lism snd lo.icily of this communly used udvcnt. Resulls showed that prc/rcatment of mate 1praRue 1)awlcy r.ls with the itmhrcen phcrK+barlrild (1'11). h1lyrhln11n.trd h1l1hrnyk /P('111, uN1, to a ntuch ksser eslent, )- I rnelhykholanthrene (MC), increased the metabolite escretion and shortened the lime of onset W peak eacrelion of the metabolile. On the other hand, aa Inhibitor or repressor of MFO's, such as either 2,1-dichloro-6-phenylphenoay- elhylamine or cobapous chloride, decreased Ihe metabolite e.crelion. lbese ra suhs subatantiate the invdvement of MFO's in the /n rlro me/abolism of dioaane. When the relNionship between the metabolism and the acvte toxicity of dioaane was espbred, il was seen that pdiosane-2-ene was considerably more toxic than dioxane. The acute toxicity studies showed an apparent t:orrelation between the metabolism and bsicity of dioxane in PCd- or MG pretrealed rats. However. PB pretreatment had no effect on toxicity In spite of substantially increasing dioaane i melabolism to p-dioaane-2one. Since Ibis lack of PB effect Indicates that p-diosane-2one requires further metabolism to exert ils toxic effecl, the totality of Ihese rewlls auggesls that the generation of the toaic substance from dioxane may Involve a multistep mechanism with pdio.ane-2one serving as an intermediate. Woo, Y-T., Argus, M. P. and Arcos, 1. C. Cancrr Research Je:1621-1623, 1978. Other su pport t National Cancer Institute. From the Seamen: Memorial Research Ldwralory, U. S. Public Neallh Servk. Nospital, and the Department of Medicine, Tulane University Medical Cenler, New Orkan.. RAfNOACTIVE ASSAY FOR ARYI. HYDROCARBON IIYDROXYLASE. IMPROVF.D METIIOD AND BIOLOGICAL IMPORTANCE This report describes an Isotopic AH/I assay, which meawres atl 1he snelahoiites formed during the /n vitro inarhalion of various tissues (liver, in/esline, lung, kidney) with (sN)-ben:o(.)pyrene and accurately determines lower enzymatic activities such as those found in the lung or kidney. TAe addition of Iwo volumes of a IM aqueous KOll/dimenhylsolfoaide (1!/8J; v/v) mixture to the enzymatic incubation medium, makes it possible to aetec- lively extract the unmclabolind benzo(a)pyrene in he.ane. Thus. the radio- activity remaining in the water phase represents all the me/aboti/es synthesized le vitro. While kss sensitive than the more commonly used fluorimelric method. Ihis new Iechnique, whose lower limit of sensitivity is estimated to be about 2.10 'r moks of total metaboliles formcd /ml Incubalhm medium, Is es- Iremely accurale, can measure eaRahepalie and low AIIII aclivi/y, and is not particularly photosensitive. Van Canlforl, 1., De Oraeve. 1. and Glrfrn, !. F,. eiochemtcaJ ond BJoph yrka/ Research Commynications 79( 2): S0S-f 1 Z, 1977. From the I ahoratoire dc ('himie MtdiLale, Inslitul de Pathologie. Sart lilman par 1.it2e. Belarum. 12 1 )

COMPARISON OF ARYL HYDROCARBON HYDROXYLASE INDUCTION IN CULTURED BLOOD LYMPHOCYTF.S AND PULMONARY MACROPHAOES In this study. !n vitro ioducibility of aryl hydrocarbon hydroaylase (AHH) in freshly lavaged pulmonary alveolar aucrophaRea (PAM) was compared to the degree of induction of Atilt in cultured lymphocytes from 13 smokers and t nonsmokers with a.ariety of noooeoplaMie lung disenes. Values obtained for Atilt in PAMs and in lymphocytes from patients withio the rwnsmoker and smoker groups were similar. Levais of AHH in PAMa freshly obtained by lavare were higher for smoken thao for rwnsmoken. With PAMs from sonamokers, bvels of Atilt were virtually Identical for celts cultured for 24 hrs. with so inducer in the medium and for cells freshly lavaged from the lun6 In eontrut, values for PAMs from anakers were reduced to about half the original kvels after culture (or 24 hn. with no inducer in the medium. Atilt values were similar when noninduced enzyme levels were compared for nonsmoker or smoker PAMs and lymphocytes. Also, when PAMs and lympho- cytes from smokers or nonsmokers were cultured In the presence of BA, similar enzyme induction was observed. Results from these studieo indicate that Atilt levels in one cell type can accurately reflect levels measured in the other cell type. Howevet, cigarette smoking dfeets the noninduced as well as the BA-induced levels of the eozyme. This is the first dme that celh (rom two scparsle tissues from Individusl subjecti have been shown to possess similar Atilt Induction capabilities when tested under Identical tulture conditions. By measuring Atilt in two separate tiawes from the same irdividual, probkms previously encountered in using a single tissue for analysis of an individual'a Atilt characterislics might he resolved. This added dimension for the analysis of Atilt in individuals might also provide reaearchers with another tool for evaluating the relationship of Atilt to chemical carcinotenesis. Mcl.emore, T. L., Afarrin, R. R., Toppell, K. L., Busbee, D. L, and Cantrell, 1'. T. The /orrnal of CNnlcaf Inrttrltarlon 60:1017-1021, 1977. Ot6er support: Nationa11ns1itutesof Health. From the Department of Medicine and the Department of Microb:iloty and Immunology. Baylor College of Medicine. Houston; and the Department of BioNogical Sciences. North Tezas Stak Univenity, Denton. ANALYSIS OF ARYI. HYDR(X'ARBON NYDROXYI.ASN A('1'1% 11 Y IN 11l/MAN l.l/N(3 TISSUE. PULMONARY MAC'ROPIIA(iES ANI) B1.CX)1) t.YMPHOCYT['S While variation In the degree of aryl hydrocarbon hydroaylasc (Atlli) inducil+ildy has been observed in pulmonary alveolar macrophases (PAM) and IympMrcylcs lrom diRcrrnl individusls. AHII values in these cclls have aN hrrn relaled w the A1111 s.hvory in other urlulosous titaues In this ahmJy d~r 1111/ ay i..*ty m t.c.h au-tually rrcised Iunif Wiue. Iresh PAM I i I and peripheral bl.od lymphocytes from 14 ciprette smoken (7 with untreated primary lung cancer and 7 eoncancer p.tienu), was measured fluorometrically. Individuals without cancer showed a good correlation (r = 0.973, p<.001) between the PAM's AHH levels and the AIIH inducibility (ezprea.ed as lob induction) in their cultured, mitogen-atirnulated lymphocytea. In the canox patient., however, these values were diaocialed. In addition, the fresh lung tissue Atilt levels and the cultured lymphocytes' fold-inductioo ratioe eorre- lated positively in noncancer patients but oot in the cancer patients. 7lhere was close agreement between fresh lung tissue Atilt and fresh PAM from iodi- vidual cancer-(roe palients, but theae values were only weakly correlated in those with cancer. Upon simuluneous comparison of Atilt activity in fresh PAM and in fresh lung tissue, and AHII inducibility in cultured lymphocytn, an ezcelknt rclalionship was found between these values in all three tissues for individual noncancer paliems (r = 0.987, p<.001). Ilowevcr, thc.e did not correlate in Individual lung cancer patients According to the data, while the capacity for Atilt induction in eaneer-free cigarette smokers is similar in the tiuues qudied. h+nR cancer patients do not have positively correlated Atilt valuea in these tiawes. Mcl.emore, T. L., Afan/n. R. R., Pickard, L. R., Springer. R. R.. Wray, N. P., Toppell, K. L., Matwz, K. L., Guinn, 0. A., Cantrell, @. T., and Busbee, D. L. Cancrr 11(6):2292-2700, 1978. OtAer wpportr National Institutea of Ilealth, National Cancer Institute and the American Cancer Society. From the Deparimenb of Medicine and S1srRery, Baylor College of Medicir and Veterans Adminiuration Ho.pilal, Houston; and the 1)epartmeot of Biological Sciences, North Team State Univenity, Denton. DISSOCIATION BETWEEN ARIL HYDROCARBON IIYDROXYLASH ACTIVITY IN CULTURED PULMONARY MACROPHAGES AND BLOOD LYMPIIOCYTES FROM LUNO CANCER PATIQNTS Lymphocytes and pulmonary alveolar maeropha6es ( PAM ) used foe evaluation of aryl hydrocarbon hydroaylase (AHII) induction were oblalned trorn 13 noncancer and 14 primary lung cancer patients. Fnzyme levels wera mcawred in cells cultured with or withoul the inducer benzanlhraeene (BA). 7Le mean levels of AIIH showed no diQerence belween noncancer and caawr patients for either noninduced or BA-induced cells. Absolute kvels and foid induction of Alll1 in PAM and lymphocyka from Individual noncancer patients were positively correlated. Ilowever, comparison of these values i. PAM and lymphocytes frrom individual lung cancer p.tients demonstrated no positive correlation. In further study, comparison of enzyme activity io macro- phages fr~eahly lavaged from the lung and BA-induced activity in cultured PAM revealed a positive correlation (or both noncancer and lung cancer p.- licnts. Similarly, comparison of eozyme activity in fresh macrophagcs and told induction values in cultured PAM was also well correlated frw M.th groups of patients From these sludies. it would appear that messuiement of Atilt In more than one tissue obtained on the sanme day from a given individual may 1A . IS

dialinguish diiferences between noncanaer nd lung cancer patients. In non- cancer patients, there is a good agreement between values for Alllt in fresh PAM and cultured lymphocytea; however, when similar menurements are compared in individual lung cancer patients, the AHH values in cells from the two diRereot tissues are dissociated. Since this diuociation between AIIlI levels is consistently found in lung cancer patients, a comparison between Alilt kvels in cultured PAM and lymphocytes could be of diagnostic aid when lung cancer is suspected. Mcl.emore, T. 1.., Alarrin, R. A.. Wray, N. P.. Cantrell, E. T., and Busbee, t). 1.. Cancer Research 38 (11) :)d01-le 11, 1978. Ot6er awrrorlr National Inslilutea of Health, American Cancer Society and the Veterans Adndnis/ralion Hoapilal, Houston. From the (kp.nments of Medicine and Microbiology and Immunology, Bay- lor College of Medicine, and Veterans Administration Hospital. Houston; the Deparlmenl of Biological Sciences and the Genetka Center, North Texas Slate Universily, Denton• and the Department of Pharmacobp, Texas College of Osteopathic Medicine, Fort Worth. EFFFCTS OF SNLFNIUM ON ARYI. IIYDROCARBON HYDROXYI.ASE ACTIVI"IY IN CULTURNI) IIUMAN LYMPHOCYTES This paper examines the effects of aeknium (Se) on induction of aryl hydrocarbon hydrosylaae ( AI1H ) activity in lymphocyte cultures and on the enzyme activity itsetf. Esperimen1a11y, human lymphocytes were cultured both In the presence and absence of induun of AHH. The presence of 10' M Se during the IaN 24 hrs. of culture (corresponding to the time when inducer was present) or during the entire culture period had no inhibitory eRect on AIIH activity in eilber eoninduced or Induud cells. However, the presence of Se in the assay ilself inhibited AHH activity by more than 50%. This level of inhiAition was seen at concentrations of the aubslrate benzo(alpyrene of 1, 3, 10, and 100 pm and Se concentrations of 0.1. 0.), 1, and 10mM, respectively. Interestingly enougA, when AI(It activity was measured In noninduced celis in the presence of low concentrations of Se, the All/t activity was slightly higher than when Se was absent. These findings Indicate that the inhibitory cfkcl of Se on tumorigenesis in• sludies with several experimental animal sya- lems may he explained in part by the inhibition of microsomal osidases re- spomibk for metabolic conversion of precarcinogens to ullimate, ekclrophilic earcinogens. Rasco. M. A.• lacobs, M. M. and OriBn, A. C. Cancer Lrtttrs ):29S-101, 1977. Other aupportr National Cancer Institute. Frum the thparlmcws of Biology and Bachemistry, Ihe llniversity of Tctas Sy.tem ('ancer ('entcr, M 1) Anderson llospital and Tumor Inslilute, IIoNn1on. OALACTOSYL TRANSFERASE IN BENIGN AND NEOPLASTIC HUMAN BI.ADIJeR MUCOSA ~ Uridine S'-diphoaphale Ralaclose:Rlycoprottin plac/osyl Iraesferaae (Oal- X), an enzyme which is involved in the bioaynlhesis of complex carbohydrates of cell membranes, has been implicated in intercellular recoRnilion, adhesion and di/ferentiation, and associated with some human endodermally derived tumon. In the present study of the activity of Gal-X in human transitional cell carcirwma, the homojenales of four established tranailional cell carcinoma lines (MG11-1, MOII-2, RT-1, and T-24) were shown to have signifkanUy increased (3al-X activities as compared with human Abroblasts in tissue culture. Because of thia, pal-X activity was then studied in fresh specimem of benign. Irdlamed, and neoplwic human bladder epithelium. Reaulu showed that 40 waya on transitional cell carcirsonsa, both invasive and raninvaaive, ranged from 24.4 to 180.0 cpm/rR of protein (mean 70.)), and SO uuya on normal or inflamed mucosa ranged (rom 0.0 to 46.1 cpm/r= (mean 9.00). 7he nsa- tority of benign mucosal specimens showed aorne inflammatory chanRea, but that did not increase the level of Oa1-X activity. These results, which show that human transiliond cell carcinoma has increased (ial-X activity regardless of tumor staRe or grade as compared with benign transitional cell epilhelium, Indicate that this increase may be useful in the development of diagnostic /luoroimmune and radioimmune techniques to ddect transitional cell carcinoma. Ilagen-Cook, K., Prout, O. R.. Ir., Plotkin, O. M., Oilbert, S. 1.., and WolJ G. Surgical Forum 29:627-629, 1979. Other aurportt National Cancer In.tilule. From the Urological Service. Massachusetts General Hospital, and the Depart- ment of Sur`ery, Harvard Medical School, Boston; and the Departmeot of Nutrition and Food Scienct, Massachusetts Imtilute of Technolop, CambridRe. I CYCLOPI/OSPHAMIDE-INDUCED ONCOOENIC TRANSFORMATION. CHROMOSOMAL BREAKACIE, AND SISTER CHROMATID EXCIIANGE FOLLOWING MICROSOMAL ACTIVATION Cancer chemotherapeutic agenta may be unequaled for the purpose of • assessing short-Ierm mammalian tissue culture assay aystems for their actual ability to predict human risk of developing cancer as a result of exposure to certain compounds. ihia paper illustrates that while cyclophosphamide does not produce oncogenic transformation in the absence of a metabolic activation aystem• sienifkant transformation does take place in Ihe presence of an exo2tneous activation source. Cell culture with and without an esogrnous liver metabolic activation system was used to investigate the eR.ct of this chemotherapeutic agent on oncogenic transformation and ehrorno.ord.l damage, including increased sister chrrKnali) exchanges. As noted above• cyclopho.- peamid- did not produce any abnormalities in the absence of metabolic ac- 17 16

tlvation, but incorporation of the activating system into the assays kd to significant tranaformalion, chroawsomal breaka and increasea in sister chroma- tid e.chantes. lhe removal of glucose-6-phoaphate nd nicotinamide adenine dinuckolide phosphate from the metabolic generating system, however, com- pktcly eliminated these aberrations. According to these retults, it may be necessary to incorporate some activation procedure into any suudy utilizing in vitro systems to determine the potential human hazards of a particular ehemical. Benedfcr, W. F., Banerjoe, A. and Ve.ka/eaan, N. Cancer Research 711:2922-2921, 1972. Prom the Division of Itematobpr-0noo/oRy. Department of Med'v:ine, Chil- dren's Ilospital of Lo. Anaeks; and the Department of Pedlalrics. University of Sixdhern California School of Medkln., Loa Angelea. MIITAGENICITY OF CAN('FR ('HPMOTHERAPEUTIC AGENTS IN 7IIL+ SALM(1NF.t.LA/MICROSOM@ TFST Many short term assays have hoco developed recently to screen potential chemical carcinotrns/mulatens, and cancer chemotherapeutic drugs have been considered partrculaily suitable for esamining the relationships among short term in vbro aaays, in vlvo carcinogenKity studies and chemical carcino- genesis In man. In the present study, 17 cancer chemolherapeutic agents were tested for their ability to mutate Salmoneffa tyoAimurirm lesler strains in the Sa/monrfla/mkrosome mutagenkity test. Results showed a high correlation between the mulagenicity and carcinogenicity of a given agent. Carcinogens positive in the test were adriamycin, daunomycin, 1-propanol-J,7%iminodime- Ihanesulfonate, cyclophosphamide, isophosphamide, hycanthone, chlornaphazin, nitrogen muMard, uraeil mustard, melphalan, and thio-TEPA. Two known earcinogens, aclinomycin D and bkomycin, were not delected as mutagena. 'i1The presumptive noncarci"cn, methotre><ate, was neitative in the lest. How- ever, tilorone and 6-mercaptopurine, tentatively clanifkd as noncarcinogena, were mWagenie. In summary, these Icau showed that 11 of 13 of the chemo- lherapeutie agents that are carcinogenic were also mutagenic in the Safmonelfa/ microwme system. In addition to being carcinogenic in animal studies, some of these agents are suspected of being carcinogenic in man. Benedict. W. F. .r at. Cancer Reua.cb J7:2209-2217, 1977. OtAer support: National Cancer Institute. From rhe f)rvisa.n of 1lemslnlogy and t)ncoloty- tkpartmenl of Medicine, ('hdJren's I/ttipiul, I 0. AnRrlcs, and the Ihpartment o1 IliiKhcmistry, t)ni- vcr%dy ol l.h/ouu., IkArl. v I CEI L CYCLE-SPECIFIC ONCOOENIC TRANSFORMATION OF CI1f/ 10TIA CI.ONE ff MOUSE EMBRYO CELLS BY I-/9-D-ARABINOFU RANOSYLCYTOSINE The drug 1-/l-D-arabinoluranosykytoarse (araC), which Ia widely used as a ehemotherapeulie agen1, is a powerful DNA inhibitor. In mamrnalias cells, it is phosphorylated to the tripho.phate level and subsequently inhibiu replication synthesis in preference to repair synthesis. Earlier studies by various investigalon show that it can also be incorporaled, although not ea- tensively, into DNA. that it produces cell eyck-apeciflc chrornatid breakage, induces chromcraoal abberations, is mutadenic to mammalian cclb, and interferes with the synthesis of gIyeoproleina and filycolipid.. It is also capable of causing malignant tranafo.mation in cultured hamster embryo aBa and in rat cells. Thia report shows that the nuclooaide also inducea oncoscnk transformation In 1he C7H/ IOTK CLtI line of mouse embryo cells and that this Iramformation is primarily S-phaae specific. Cell lines derived from type 111 transformed foci grew in soft agarose and produced tumors in immuno- suppressed aynneneie mice. In cells synchronized by poatconfluent growth inhibition or isokucina deprivalion, transformation was cell cycle dependent. Maaimal tramformation was obiervcd in cells treated In the S phase, although tome transformation occurred in cells treated in the O, phaae. The authors indicate that some of Uese reaulu may be eaplained by the facl that araC is capable of causing ehroanoaornal lesions in the (l, phase and of delaying the entry of O, cells inlo the S phase. They aiso.uggest that specific chromo- aomal ehanges may be particularly important in chemically induced malignant transformation, especially since the eapresaion of cancer in cells transformed by ara-C is associated with specific ehromoaoroal imbalanua. lonea, P. A., Baker, M. S., Bertram, l. S., and Oewedict, W. F. Cancer Re.eareA 77:2214-2217, 1977. . Other a.rpport: National Cancer Institute. From the Division of Hemalology-(Mcolop. Department of Medklne, QiY- dren: Hospilal, l.os Angeks; and the Department of Eaperimental Therapeu- tics, arace Cancer Drug Center, Roswell Park Memorial Institule, BuAab. SPECIFICIIY OF HUMAN. RAT AND MOUSE SKIN EPOXIDtE HYDRATASIi TOWARDS K-RF.OION EPOXIDES OF PULYCY('I.IC HYDROCARBONS Micro.omal enzymes have, the capacity to transfonn aromatic and okbnic compounds (ncludrng benzo/w/pyrene to eposides which are elec- trophilic and can apontaneously form covalent bonds with the nucleophilie moieties of tissue 1)NA. RNA and protein. Several studies have shown that epoaide hyJratase has a unique role in the "tabolic formation of carcino- gens from polycyclic hydrocarhons. llris repixt emrmeralea avme of Ihis enzyme's properties in human, rat and mouse skin mirrnsomal fractrons and compares them to those found in liver preparations 7he skin enrymc hydrated all epoawka tested, with human fraclions showing the most activtty, followed IB • 19

in descending order by mouse and rat samples. The level of activity, however, changed with each of the various polycyclic hydrocarbon. as substrates. In human skin microsomal fractions, the eposide hydratue activity dwwed little p11 dependence and was inhibited by small molecular weight inhibiiors in a manner similar to that of the liver microsomal enzyme. However. tM! activity does seem to vary considerably among individuals, as demonstrated by the skin microsomal fractions of sin humao subjects. Thu variation was not due to skin disease or Ireatment and seemed indepe.dent of age or aes. Tlu enzyme activity in abdominal skin sampks. Iwwever, scemed to be lower Ihao in samples taken from the leg or breaM, suggesting a possible correlation between the activity level and the tissue's sile of origin. According to the re wlts, the enzyme found in skin microsomal (ractiona is Inlrinaically similar lo dut found in liver micro.omal fractiona, so that any data obtained with liver epotide hydralase are mosl probably also applicabla to ski.. Orirh, F., Scbmaaamann. H. and Bentley. P. Biochemical Pharmardogy 27:17-20. 1978. From the Institute of Pharmacology. The University of Mainz, Mainz, West (;ermany. THC ROl f? OF GIiNF:TICS AND HOST FACTORS IN CANCER SUS('EP I IBILTIY AND CANCER RFSISTANCE 7his paper provides a survey of current findings relating to cancer susceptibility and resistance in humans and attempts to apply this knowledge in a meaningful way to clinical medicine in general and to cancer control in particular. Familial cancer distribution In man, geographic variations in cancer susceptibility and tissue susceplibility in hereditary cancer are esamined here as pertinent variables in the overall cancer problem. Such eaampka as carcinoma of lhe brent, colon, von Recklin=hausen's neurofibromatosis, and multiple primary malignant rseoplasms emphasize Ihese variables. One of the most consistent observations in cancer genetiea is the significant eacess of mullipk primary malignant neoplasms in cancer prone families. Problems in genelic heterogeneity and mechanisms involved In biological markera in cancer re carefully considered. While many esampks of cancer suaceptibility are presented for eaansinalion, the likrature contains few inslances of cancer re- siuance. This is unforlunate, since a beller understanding of human cancer resistance might kad to the elucidation of inechanisms which could protect against cancer, inslcad of merely controlling the disease after its onset l.ynrh, It. T. rf al. Canrrr nnrr•rion and Prrvrnrion 1( 1):173-224, 1976. Othrr support: Narw+nal In.ututn of Ilea)rh and the Fralernal Order of 1'ig1e% 1 r..in rhe 14p.rrn.rnr ot 1'rrveMive MrdMrne and Public Fledrh, ('rei`hlon llniVrrvl~ 1.h.rd .-1 A(r.hin.r, t)nuha I GENETIC AND COMMUNICABLE EFFF.C,T$ ON CARCINOEMBRYONIC ANTIGEN EXPRESSIVITY IN THE CANCER FAMILY SYNDROME Carcinoembryonic antigen (CEA ), which hr .hown potential as a mut.r for endodermal cancer (particularly cancer of the colon), was studied her. lo: (a) leat CEA's potential as a discrimioanl of penetie cancer risk in canotr- prone families: (b) quantitate the role of age and duration of cigarette smoking in CEA esprea.bn; and (c) test for connubial effects on CEA is the hereditary cancer family syndrome. To do thia, plasma CEA /erela wera ezamined in 198 relativea and 84 spou.ei of ais carsrxr-prosse kindreda, and in 191 normal controls. The CEA di.tribulion. per as were transformed to square root CEA (VCEA) as a correction for skewness and kurtosis. The snean VCI'A did not differ betwoen cancer family members and controls of the same smoking status. Funhermor., the ratea of change In V('P.A with increasing age and duration of smoking were similar in both cohorts. Ilow- ever, when pedigree data were classified by closeness of relationship to canocr patients there was a siRniflcanl lintar increase in mean VCEA with increasing jenelic risk. Surprisindy. CEA kveb of spouses showed the sanre treod as their direct-genelic-line malea (spouses were classified according to the genetic risk status of their mates rather tha. their own genetic lineage). Among both direct-line relatives and their apouw, there was a consistent linear decline in \/CEA with declining risk Na/w. Inlraelasa eorrelatian of CEA between direct-line relatives and their npouse. approached ,ignilkanoa when both spouses were concordant for smoking status. These data reflect as intricate chain of events htvolving Rendia, eomnwnicability, and physical interactin4 phenonsena. An integrative target cell theory ia proposed here so tsplain CEA espressivity in this syndrome. In the given rnodel, the larlet faclors appear to Interact differeatially at the nsolecular-cellular level to caur CEA varialions. GrlrtL, H. A.. LyncA, H. T., Harris, R. E., and Vandevoorsfe,l. P. Cancer Research 3t({):2527-2521, 1970. Other aupprt: HoQmann-LRoche, Inc., National Cancer Institute. FannM E. Ripple Foundation, and the Fraternal Order of Failkt+. From the Dewrtment of Preventive Medicine and Public Heallh, Creighton University School of Medicine. Omaha; and the Department of Immunobp, HuRmann-LaRoche, Ine., Nutky, N. 1. CARCINOhMBRYONIC ANTI(7F.N (CEA) IN T/IP. CANCER FAMILY SYNDRJ)ME 0 The carcinoemhryonie antigen (CEA) has been described as a marktr for the detection of endodermal cancer, especially malienancy of the colon. This study attempts to furlher elucidate the potential of CEA as a discriminant of cancer risk in kindreds demonstrating ahe Cancer Family Syndrome. Plasma samples were collected from 143 relalives and 65 spouses of sia csncer-prono kindreds, over a pcriud of four years. For statistical eompariaons, rclatives were classified as: (1) cancer patients. (2) unsffec/ed Individuals at high genetic risk (one or more first degree rclatives d(eeted), and (3) unaffected individuals 20 21

at low genetic cancer risk (no Ant-depee rslalives with cancer). Unrelated spouses were assigned the same clan number as their direct-line mate. In cancer patients and relatives at high genetic risk, the mean \/CPA was signifkantly higher than in relatives at low genetic risk. Surprisingly, how- ever, unrelated spouses had mean levels of Y/CPA similar to those in the corresponding risk clau of their direct-line mates. These data suggest that both a genetic and a connubial factor operale on CP-A, the latler presumably traceable to a common environmental aRed actinR in concert with the desrte of genetic predisposition to oncolienesis in this syndrome. These resuits por- tend a hypothesis bascd upon a corntnunkabk-jenelic Interaction involving transformed viruses that oAen a partial esplanation for the genesis and trans- miuion of oncodevelopnuntal prolebr is memben of high cancer risk k indreds. OrLr4, H. A., Lynch. H. T., Ilarris, R. H., and Vandavoorde, 1. P. Canctr I)()):1374-1S7f1, 1978. Other eupportr IloRmann-lARoche, Inc., Roeha Research Center, the Na- tional Cancer Instilute, the Fraternal Order of PaRles and the Fannie E. Rippel Foundation. From the Department of Preventive Medicine/Publie Health. Creighton Uni- vcnity School of Medicine, Omaha; and the Research Division, Depatlrxnt of Immunology. HoRmann-laRoche, Inc., Nutky, N. J. OPNPTIC REOUT ATION OF Sl1SCPPTIBII.f TY TO POLYCYCLIC-IIYDR(X'ARBON INDl1CFI) TUMORS IN THE MObSP The eytochrome P-ISO mediated monoosygenases are a collective ea- ample of Phase-I enzymes which can be regulated in the mouse esperirnental system. These membrane-bound mullkomponent enzyme systems, while re- yuirinR molecular osygen and NADPH, are tnown to metabolize pulycyclk hydrocarhons such as bcnzo[elpyrene (BP), )-melhykholanthrene (MC), biphenyl, elc. While it is now generally accepted that during the detosifkation of these compounds by Phase-I and Phase-11 enzymes, polent reactive inter- mediales, capable of binding covalently with cellular macroreoleeules, can be formed, the association of steady-s/ale levels of certain of these reactive (ntermediates with cancer, mulation, and toakity is presently of great rtsearch Interest. Primarily, the aryl hydrocarbon hydroaylaae (AIIiI) assay Is a reliabk, simple and very sensitive assessment of aromatk hydrocarbon responsiveness following trealment of animals with polycyclic hydrocarbon ieducen Pspcri- mentally, with the use of inbred strains of mice and the appropriale genetic crosses, it was possible to demonstrate that a single al/elk difference or a very smaM number of genes could have a profound Influence on n individual's increased tusceptibility to chemical eareinosenesis, drug toskity, and con- genital maQormate.ns. Ilowever, it was also shown that genes other than Ihe AA locus miRhl influrnce an individual's risk (or lumorigenetis. Aside from the Ah hxus, rhree miNhfyint factors in tumorisenesn Ihat may be of im- p.nlarKe and m.y al+n he undcr Renetic control inchtale ONA repair, vital t.l~tcsau.n. arnl nnrnnnnl,.graa t,.lcr.rnac Ihe inlerpl.y bttween Ihe Ab Inctn and these above.nentioned factors, as well r the role of these factors by Uhemselves, in susceptibility to cancer ara currently being atudied. Kouri, R. P. and Nebert, D. W. (/llkro6lolog/caf Aisoclates, Inc.) lo: Hiatt, 11. Ii., Wataon, I. D. and Winsten, 1. A. (eds.)' OrlSfiar o/ Hrrnan Cancer: Book B Mrchanlinu o/ CanclnoRenefi,r, Cold Spring Harbor Confer- ences on Cell Prdiferation, Cold Spring Harbor, N. Y.: Cold Spring Harbor Laboratory, 1977, vol. 1, pp. ta 11-fi13. Other arpportr National Cancer Institute. , Pnom the Dep.rtment of Biochemical Oncolopy, MkrobiobRkiql Arociates, Ine., aod the Developmental Phartnacolory Branth, National Institule of Child Health and Human Ikvelopment, Bethesda, Md. 2,),7,fi-TETRACHIARODIBPNZO-r-DIOXIN AS COCARCINOOEN CAUSINO )-MPTHYId'HOLANTIIRPNGINIT7ATP.D SUt1CUTANEAUS TUMORS IN MICE OENEI'ICALLY `NON-RESPONSIVE' AT AH LOCUS There it growing evidence that a prereaui.ile for mutaReania and earcinorneais by many. it not aB, polycyclie hydrocarbona is metabolism to neactive intermediates by rytochrome P-4S0-soedialed rrronooaygenaw wrch as aryl hydrocarbon hydroaylaae (AHH). 2,7,7,8-tetrachlorodibenzo-pdbain (TCDD); which is a potent Inducer of AHH activity and Pr4SO in both responaiw and nonrespomive mtoe, was us.d In this study to daterruina whether it aould enhance Ihe .uaceptibility rot C37BL/6 (responsive) and DBA/2 (nonrtaponsive) mice to 3-methykholanlhrene (MCA)'induced can- cers. Eaperimenlally, the possible effect of TCDD on four hepatic enzyme acdvilies. AHH, epoaide hydrae, Rluuthione S-traesferase, and uridine di- phosphate-Rlucuronoayltransferue, were studied. la the liver of either Nrain of mice, 12 to It hours after a high dose of TCDD, the magnitude of In- erease in Allli was many times greater than any change in any of the other enzyme systems. At an i.p. dose that killed 30 to 70% of the animals in sach group of DBA/2 or C37B1/6 mice, TCDD e.wed no lumors in the sur- vivors 36 weeks later. TCDD given I.p. to C3781J6 mice two days prior to, or simuUaneously with, s.c. MCA did no1 dfeet the MCA carcinogenic Indes, while i.p. TCDD Riven b DBA/2 mioe two daya prior /o s.e. MCA incrcased 'N slightly. When s.e. TCDD and s.c. MCA were given sfewllaoeoualy in the same vehicle to DBA/2 mice, Ihe carcinogenic Index Increased markedly. These results Indkate that TCDD is not earcinoRenk in this lest system. However, k docs appear to be a eocarcinoten that might act by indudng AHII activity and its associated P,-4S0 at the ake of inoculation of MCA in genetically noruaponsive mice, thereby resultinR in more eflkient metabolic conversion of MCA to the prosimN or ullimate carcinogen. Kouri, R. E. ef..f. (MkroAtolotkat Assoclater, lne.) Cancer Rtsercil )t<:2777-27ta), 1978. Prom the Deparlment of Biochemical (McoloQr, Mkrolrloloolicsl:Associate., Inc.; the l.abora/ory of ('htmnlry. National Institute of I,rrhtiris. Mclabarlism, and Digestive IHseases; and the Developmental Pharmacology Branch, Na- tional Institute of (:'hiW Ilealth and Human Ikvelopment, Betheda, Md. 22 23

1 COMPARISON OF THP EFFECTS OP BEESWAX:T7t1OCTANOIN AND T RIOCTANOIN VEHICLES ON 3-METHYLCHOLAN INRENE, BI:NZOIaJPYRENE, AND 7,12-DIMETf1YLBENZ(aIANTIIRACENE SUBCt1TANEOUS CAR('INOOENE.SIS IN TIIREE STRAINS OF MICE AND ONE HYBRID In carcinogenesis studies, aolubilization of a suspension introduced into warm becswas to form test implants tor asisnal models determines both the dose that can be administered and the doss available to the individual tarilN cells at any point. The present experiment attempts to determine the relative carcinogenic effecu of three diHerent aubeutaneous doses of 3-mahylcholan- threne (MCA), benzo(alpyrene (BP), and 7,12dimethylbenz(alanthracene (DMBA) suspended in Iriuclanoin, as compared 1o a 1:1 beeswaa (B): Irioctanoin (T) vehicle. Three inbred snd one hybrid mouse atraina were used in order to determine if tunar induction was relaled to a ei/ference in 1he ability of the straina to solubilize and metabolize material from the pellet compared to their ability to metabolize the qrcinogens when sdministered In a soluble form in Irioctanoita. Median tunwr doae kvels were ai6nifksntty higher when the three earcinogem were suspended in trioc/nwin. With B:T, the three carcinogens differed in their abilities to be absorbed or solubilized from the vehicle by the various strains. Is C3H/Aat mice, BP in B:T failed to produce tumon; so did MCA. BP, .nd DMBA with II:T in BC3F, mice. Neither were any tumon induced in the C37BL/6 tlralo by DMBA or MCA la B:T, or in DBA/2 animals by BP or MCA in B:T. These data iadicate that the interpretation of lumor induction results obtained with B:T vehicle may be related to the conditiorn of bioassay rather than to the earcinoger.ic potential of a compound Whitmire, C. E. and Lopez. A. (M(croblologkat Asrociarei, Inc.) Jor.nd o/ dhe Narlonal Cancer /nstirrre 61(1) :1107-I 111, 1978. From the Department of Experimental OneoloBy, Microbiological Associates. Ine., Bethesda, Md. INDUCTION OF TYPE C RNA VIRUS FROM CULTURED RABBIT LYMPIIOSARCOMA CELLS Type C RNA viruses have been Implicated in the etiolop of lympho- hematopoietie tumors. Consequenlly, there have been extensive attempts to kwhte infedioua type C viruaes from species in which these neoplasma occur. Recently, type C viral Information (p30 and RNA-directed DNA polymeras. /RDDP/), but not complete infectious virus, was demanstrated in W11/1 rabbit spontaneous lymphosarcornaa. The data presenled here ahow that hak+itenated pyrimidine (S'-iodo-2'-deosyurine. or IdUrd) induces the release of type C RNA virus partick(s) containing an RDDP and the p39 structural prolein, which, although they appear to posaen unique snligenic delerminants, share sonx in common with other type C virvses. Hybridizdion between rabbit virus DNA and DNA estracled from normal rabbit liver revealed cellular DNA wrnh ntnletthJe sryuentcs h..mulugtan lo Ilwtse rd all laheled cnmplcmentary I/N 1 U.u.. nIN .•/ the t.bl.,t vir.l t.n..mc 11w+. twNh mtrm.l rahhd liver I)N 1•rn.l 11N1 I...u. uh.nr. ..mlun nu. kuhdc sryucut cf related I to the rabbit virur, indicalin8 that the lalter is truly endoperww and not a contaminant. To prove that this Is Mdeed w, the virus muat now be isolated, characterized and compared with othtr type C viruses as well u with cellular DNAs. To this end, a cell N.e which will allow for the replication of tlr induced virus is being sought. Bedittian, H. U., Foa, R. R. and Afeier, N. Jorrnaf ol Ylroloty 27(2):313-319, 1978. Other support: U. S. Public Health Service. From The Jackson l.aboralory, Bar Harbor, Me. TRACIIPABRONCIIIAI. CYTOLOOIC CHANOPS IN MAI.IONANr MELANOMA Hisliorytes counted in smean made from the tracheobronchial waahiap of 208 patients with maliinant melanomas were found to be ).12 times more rlwmerou. 11un those in a control group matched by ses. age (decades) and smokal8 habit butt without any type of prediaBnosed malignant lesion. Since 52 of the patienu with melananas had been followed up for period. up to 40 eantlu after surfiery, a further attempt was made to coaelats tha mortstily with the number of bisllorytes found in smears during operations. Of these 52 patieet., 40 were still alive. Among the 12 patients who had died. d had histiocyle coww below 649 (the mean number for the mclanorea series) and six above that oount, or a high 1o low count ratio of 30%, whils 30 out of the 40 patieeb who bad survived Md high hisliocyte counu, or a high to low ratio of 75%. The difierenoe in ratios belw«n the two aeeties was of borderline statistical signiBc.ace. The large percentage of high tracheo- bronchial bistiocyte oounb in the survivor group probably indicates a atroq immune cellular respoene, which may explain the Nth survival nte. Tbers fore, since morlalNy depends on a variety of facton, these findinp sn presented simply as a method of asus.in8 boat response in malignant diuw. C)haton. 1. aI. aJ. Archives o/ Pathology asd Laboratory Medlci.e 102:631-631, 1978. Prom the Department of Aneslhesiolop, New York Univessity Medical Cenler, New York. DIAONO.4TIC AND PROONOSTIC. SIONIFICANCE OF TRA('HI?OBRON('HIAI. EPI111P.1_IAI. MULTINUCLNAT7ON hor this cylololai study, traobronchid washin were obtained from /,910 eonsenlin~ patienlsic of bolh sesesiheabout to under~op~encral endolracheat anesthesia for surgery. One half of the total group had malignant disease and the other half, matched by sea, age (decades) and smoking habUa but with- out malignancies, served as eontrola. Besulls showed that the mean percentage of mullinuckaled cells (pmnc) in smesn from patients wilh malUgnancles and from all controls was 2 381007 and 1.42.tO07 respectivelT (P<0(NI0S). In cases were sui><lass/fied by age or sea, the ddlererke in hmnc hctwccn 2 3

BACTI?RIA1. CLF.ARANCE FROM T11E LOWER RESPIRATORY TRACT OP 7 FIL' MOUSE: EFFECTS OF ACUTE CIOARETfE SMOKE EX POSUR E Existing evidence suggests that the mucoeiliary and alveolar mscrophaje systems play  central role in hoet defense against inhaled inanimale and infectious agents. "Ihis study was undertaken to examine the eRecto of acute esposure to puffed cigarette smoke on bdt resistance to an airborne bac- terial challenge, and to assess the revctsibility of smoke-associated changes in bacterial ckarsnce. For this purpoae, simultwseorn measurements were Iaken of the numhen of viabk bacteria remaininR in the trachea and lungs of mice at various intervals after the inspiration of aerosolized Sraobylococcrs arrers In the sbsence of cigarette smoke (control group), during smoke inhalalion, and after the cesaation of smoke exposure. Rapid clearance of S. arreur from the trachea was seen after bacterial implanlatioe by an aerosol exposure sys- tem: 35% were cleared in 13 min. Sf% in 30 min. 73% in I hr. e4% in 2 hrs, and 95% in 4 hn. l.un6 ekaranw was equally eReclive, but not as rapNl: )17f% of the organisms were cleared in 13 min. 42% in 30 min. SS% in I hr. 70% in 2 hn, and 90% in 4 hn. When smoke was inhalcd, the following effects were noted on the mice: within IS min. after bacterial chal- kn`e, snwke<zpoud mice cleared 74% of the bacleria from the trachea, and control mice showed )4% clearance with a relative retention ratio of 2.5. TAis ratio was maintained with minor variation over the entire 4 hour smoke- exposure period. In contrast, esposure to cigarette smoke was accompanied by an impairrnent of lung clearance and agraded increase in bacterial reten- lioo related to the length of time the animals were exposed to smoke. Re- venibility studies, however, showed that bacterial counts from both the trachea and lunp of mice eaposed to cigarette smoke for I hr returned to the levels observed in control mice within ) hrs after the lermination of smuke exposure. These findings indicate that cigarette smoke caused a transitory acceleration of tracheal clearance and suppression of lung clearance. Craineri, l. l. In: Developments in lndustrid Mkrobblo:y, Washinston, D.C.. American Institute of Biological Sciences, 1978. vol. 19, pp. 415-426. From the Department of Pathology. Queens Hospital Center, lanb Island )ewish-Hiliside Medical Center AfRliatias, Jamaica. N. Y. POSSIBt.H MF.C)IANISMS OF P.MPIIYSP.MA IN SMOKERS. IN VITRO SUPPRF.SSION OF SERUM PLASTASN-INIIIBIt'ORY ('APACII Y BY FRI?SN ('1OARE'TIP. SMOKE ANl) 113 PREVP.NI ION BY AN7IUXIDAN IS Several recent studies suggest that emphysema in smoken may be due in part to local suppression of elastase inhibition by cigarette smoke. Now, ekctrophnresis and immunaelectrophoresis sMrw that bwNh serum clastase- Inhihitory capacity and Ipha, proleinase Inhibitor /e,Pr)/cla.rase cnmples furruatian are dccrcased by huffered, /resh cigarette snsoke intaK/,rced into aqucnus serum s-rtuiions to whrch pAncreatic elastase is then added. At the same time, the frm, active prole..e content Is increased. Introduclioe of phenolic antiosidants, particularly thymol and hydroquinone, into freshly pra- pared aqueous smoke solutions )tW before the addition of aerurn completely blocks the subsequent auppressas of serum dastase-inhibilory capacity by cigarette smoke. When the effect of substituting nsodel ostidanta for cigarette ,moke in the esperimental system was eaamined in order to further test 1he hypothesis that suppression of serum el.atase-inhibitory capacity by cigarette smoke is due to oxidation reactiorr, all three agenta used (N<hbroauccieL mide, chloramine-T, and hydrogen peroxide) reproduood the suppression of serum elastaae-inhibitory capacity observed with the soluble urwke eompa nents. These results suggest th.t this in vitro inhibitory activity may be du. to smoke-induced osidation of cysteine and/or methionine reaidues io .,Pi. Such suppression of elas/se inhibition by a,Pi In the hmb, whether caused by inhaled cigarette smoke or by environmental oaidants, could be responsible for the proteolylk dcsruc/ion of lung tissue observed In emphysema. Carp. N. and /awv0, A. American Rerkw of Reiolrarory Diieases 11/(7):617-621, 1978, OtAer supports National Heart, Lung and Blood Institute. From the Department of Patholop, State Univenity of New York at Stoay Brook, Stony Brook. (.PS FON('TIONS MftABOI.IQUPS DU POUMON. IV. PROTE.'INASPB ET ANTIPROTCINASI±S EN PATHOLOOIE PULMONAIRE This review presents the body of evidence supporting three porsble \y- paheses concerning the development of hurnas pulmonary emphysema, a.d formulales several questions that must be answered in order to perfect more sophisticated eaperimentd models which will ehscidate its patho6ene.is. The most popular hypothesis cotniden emphysema to be the result of a disrupted homeostalic equilibrium between pulmonary proteasea and antiproaeases, based on the following observations: (1) isscreaed proteinase activity in the luo6 results in emphysema; (2) endogenous enzymts can induce emphyserssatour like lesions; (l) there is a correlation between the proteinase-antiproleinasa equilibrium and the clinical evolWion of the disease; (4) pulmonary neutro phils and macrophages are activated in auch a manner that the incre.se In their number is associated with the appearance of emphysema in certain sryF tems. A second hypothesis .utilResta that an ini/i.1 prokolylic (elaNolylic) event unmasks a specific alveolar antigen which triggers a deslrucUve ins- mumrbgic response on the part of the host. Thtee major arguments favor Ihio particular concept: (1) once trillitered. Ihe alveolar destruction continue. I. the absence of any deteclahle elaslase; (2) immursologie mediators iuduce the rekase of various enzymea, including elas/se; (3) emphysema patients have elevated concentrations of circulating anticollagen antibodies. The third hypo• thesis suggests that either ezposure to environmental faclon, such as tobacco arssoke, or physiologic aging mechanisms kads to degenerative proctsaes ler. minsting in en,physema. Sunh rkgenerauon could result from ae acquired wseeplihdity of elatin to degradation, from increased neutrophilic degranu- Ialioa, or from a greater af6nily of the enzyme for its substrate. Navorinb this 28 29

last hypothesis are the facts that: (1) environmental fsclon can inlluence the degradation rate of connective tissue; (2) anUproleinasew, proteinases and the rekase of enzymes are influenced by physiologic aging as well as by environ- menld lacton responsible for the development of emphysema. Unfortunately, none of the esisting models utilizes completely homologous ekments, nansely, lunt, enzyme and inhibitors from the same speclea, and none provides a time continuum analogous to the several years required for the evolution of the patholo6ie processes in man. In addiliM the statistically demonstrated rela- tionship In man between empbysema and tobaaeo smoke has not been estab- lished In the available eaperieental ntodeM, and data concerning the relation- ships within the lung between the syolhais, rekaae, and elimination of elas- tase or its inhibitors are still IackieK. Even if the first hypothesis currently seems to best espl^in the pathogenesis of human emphysema, much additional work is needed to clarily the ensyane:lnbibitor equilibrium and the factors likely to modify it. WeinDarm. G. and Kimbel. P. fa Rrrre dr M/dtclnr 1 S()6 ):1509-1 S 11, 1977. Other supports National Heart, Lung and Blood Institute. From the Pulmonary Disease Seetioa, Albert Einstein Medical Center, Phila- delphia. DEGRADATION OF TROhOEI.ASTIN BY PROTEASES F.Iaslin, which is largely responsible for maintaining the elasticity of major blood vesseis and lung tissue, is a highly insoluble protein. However, a single polypcptide, tropoelastin (m. wt. 70,000), appears to be a soluble intermediate in the biosynthesis of the fiber. In studying the pathogenesis of chronic obstructive lung disease (emphysema), eaperimentd models have dem- onstrated a strong correlation between the production of emphysema ^nd the cleavage of soluble elastin. The fact that this process may involve the peri- odic destruction and repair of pulmonary lisaue, moreover, suggests that in- tetruplion of elaslin biosyMhesia by degradatioa of tropoelastin could play a significant role in emphysema development. As a fint step in eapbrinR this possibility, the present report describes a simple asaay capabk of detecting a single break in the Iropoelastin polypeptide ehain, and she protein's sensi- tlvily to a number of proteases. Purified radioactively labeled tropoelastin, incubated with dilute solutions of proteasea at physiologic p11 and ionic stren6th, was rapidly de6raded by a variety of these enzyrnn, includin6 kuko- cyte and pancreatic elaslase. This vulnerability of tropoelastin to proteolylic digestion even though both enzyme and substrale are cationic at neutral pll, may be parlly esplained by its probable configuration as a random coil. Be- cause even a single cleavage of the Iropoelastin chain would probably siRniB- candy diminish its ability to lorm normal elastic 8ben, any consideration of the psthogenesis of emphysema should include the potential role of any num- ber of lwptiJasus that might he rckased in the lung. ('hrrstner, P. WnnAam.n, C, Sloan, S; and Roseotdoum, 1. Anu/vnt at Mrarhrmin.r RR 6141 hNR, 1978 OI tiar ar. pport r NRtional Institutes of Heahb. From the Center for Oral Research, Department of Anatomy and Histokgy. School of Dental Medicine, University of Peorrylvania, and the Pulmonary Disease Section, Albert Einstein Medical Center. Philadelphia. PURIFICATION AND CHARACTERIZATION OP CANINE a-l-ANTIPROTEINASE During a course of studies on the etiology of eaperimental canine em physema, it became necaaary to purify canine rl-antiproleinaaa (AP) to serve as an antilieaJc and biochemical standard for diAerentialion of the vari- ow paMease inhibilors found in The dog lung. The present methodology pap,r describes the purilkation of dog AP 94fold with a 25% overall yield. The purification scheme includes anionsaehan6e ehromatography, to separate away the bulk of the serum albumin; allinity chromatography by insolubilized con- canavalin A. to remove most of the other serum proteins as well as traces of albumin; and, enatly, sizing on Sephacryl-S-200. Unique to this puriika- tioe scheme is the batch use of insolubilized hemoillobin-SepMrose beads to remove the ubiquitoua contaminant haptoglobin. Characteristica of the dog AP are the presence of at least two iaoinhibitor forms and its lack of immu- nok+gicat crosa-reaetivily with Munau AP. Abrams, W. R.. Kimbel, P. and WdwDarrn, G. Alocliemlitry 17(17):3336-3361, 1978. O/Aer wpporNr National Institulea of Health. From the Department of Medicine. Pulmonary Disease Section, Albert Eia sleia Medical Center. Philadelphia. SPECIFIC LYSINE I.AHELINO BY tsOH- DURING AI.KAI.INE CLEAVAGE OP TIIE orl-ANTITRYPSIN-711RYPSIN COMPLEX Scrine proteatN:s cleave proteiua by binding to the amino acid for wMeA they are specific and forming a series of intermediales, includinp a ktrahodral adduct, an acyl ester, and two frapmenu of The protein and the free enryerr. 11 has recenlly been suggested that the or1-anlitryprin-trypsin eomples is an acyl ester analogous to the acyl inlermediate that forms helweeq Irypsin and its substrates. This cornpka can bd split at a high pll, releasing a component of . 1-antilrypsin, which has a new earbosy-terminal lysine and is missing a peplide of about 4000 daltons. In order to determine whether the I-anti- trypsin Is bound to trypsin through this new carboay-terminal lysine, as ea- pected it the slxsve hypothesis is correct, the eomples was split In the pres- ence of 18111 . When the csrbosy terminal lysinc thus obtained was cleaved with earbosy-peplidase B. singly labcled, doubly labekd, and unlabeled lysine were recovered. '11hese data support the hypolhcsis that the ~1-anlitrypsin- 31 30

trypsin complex Is either an acyl ester or a tetrahedral precursor thnt is traru- formod into the acyl ester form at a high p11. The methods use4 in these determinatiom may prove useful in establishing which a-l-antitry fsin amino acids bind covaiently to each enzyme, in the event that other enrymes are bound by a similar mechanism. Cohen. A. Q. er .l. procrrdlntr o/ the NaNonal Academy oJ Sciences o/ the United Stotes oJ Amtrka 74(10):4311-4314, 1977. Other supportr National lieut, Lund and Blood Institute. Prom the Department of Mediciee, Temple Ueivenity Hesdth Scir nces Cen- 1er. Philadelphia; and the Department of Pharmaceutical Chemistry, School of Pharm.cy. University of Cslifornia, San Pranciaeo. IiYiP.RA(T1ON OF HUMAN Q-1-ANTITRYPSIN WT'ifl PORCINE TRYPSIN The mechanism of the interaction of human rt-antitrypsin with porcine trypsin wss examined by analysis of their reaction producls. The eRects of different molar ratins of s-l-aolitryp.io to trypsin and varying incubation times were analyzed by polyacrylamide gel ekctrophoresia. Results showed that the complct was only slightly degraded up to 24 hrs of continued in- cubation at 37'C, but degradation of the complex became more evident alter 72 hn. One catalytically active component of porcine trypsin was not In- hibited by .-1-snlarypsin 77+is component, which could account for the de- layed degradation of the compks, may be an autolytic product of the trypsin. The amino acid composition of the complex was In accord with that pre- dicted foe a 1:1 molar ratio of enzyme to inhibitor. In addirion, the compk: bore the amino-terminal residues observed for trypsin and native a-l-anti- Irypsin. The enzyme-inhibitor eompkx could be dissociated in alkaline solu- lions, with the release uf a small peptide and a large fragmenl of e-I •anti- trypsin, which had an amino-terminal threosiee residue and a molecular weight between 46,000 and 30,000. These and other findings support the hypothesis that trypsin reacts with rl-anthrypsin at • Lys-Thr bond and that no pcptides of the reactants have been lost from the inlact complex. Avail- able evidence suggests that the .-I-antilrypsin-Irypsin complex is analogous to the tetrahedral or acyl intermediates which form only transiently between trypsio and ils wbtlrates. Cohen. A. d., Otcty. D. and )ames, 11. L e/acAcmlrtry 17( 3): 392-100, 1972. Other supportr National Heart, I.ung and Blood Institute and the (3eneral Research Support Fund of the Temple University School of Medicine. Proen the hrpartnKnt of MedKrne. Temple l/niversity fle.lth Science, ('en ttt, Phd../c1pMa, (scparu.Knt of Mcdnrne, San Frarwuco (icncr.l Ituspital, arwl Pulrm.nsry Clrr ulreed t cnter al Retrarch and ('ardiovasetdar Research Inslbtule• lhnvcrvty .d ('dduro.s, San I rsnci.co I INACTIVATION OF IIUMAN a, PROTEINASQ INIIIBfiOR BY TH1O1. PROTE?INASES Human plasma s, proteinase inhibitor, alw known as .,-antitrypsin. Y the body's principal modulator of serine proteinsea (such as those released from phagocytic cells). This paper reports a careful study of the Inleraction of the two thiol proteinases, pap.ia and cathepsin B1, with a, proteinaae In- hibitor. Results showed that papain dused a rapid decrease in 1he activity of ., proteinale inhibitor. Examination of samples of the incubation mixture showed a shift in the molecular weight of the inhibitor from 13,000 td 47,000, indicating peptide-bond ciewage during the incubation without the formation of a stable complex. Apin, when cathepsin BI was incubated with a, pro- tinae inhibitor, the activity of ths Inhibitor was lod. However, the reaction was signiflcantly different in Ihat the interaction, cleavage of peptide bond(s), and inactivation of the Inhibitor were sloichiometrk. These resulu su4pat a new pathway by which active-inhibitor eo.eentratioos in tisaues may be sub- stantially decreased so that endogenous proteolysia may occur. That is, any type of reactioa which might activate thiol proteinasn could result in the inactivation of normally protective proteinse inhibitors. lohnsoe. D. and Travis. J. TAe eioc/irmJcd Journal 163 ( 3):639-641, 1977. Otber supports National Institutes of Health. From the Department of Biochemistry, The University of Oeorgia. Atherr; and the Tissue Physiology Deparlment, Strangeways Research Laboratory, Cambridge, England. METABOI.IC ACTIVITY OF DBVELOPINO RABBIT 1-UNO The uptake and metabolism of 10111 kuciue, (U-'4C) glucose and 1'11) palrnitate were studied in late fNal, aewbwo, and developing rabbits ,aid ootsr pared to that of adult and old (1-1.5 yean) animals. Although lue6 com- position did not change markedly, lung metabolism showed dreat diAereooa at various developmental periods. Ltucine iocorporation inlo protein was highest in fetal and neonatal luap, and decreased 30% during /ho flrst week after birth. Patmitic acid itecorporatiou decreased during the 1lrri week at/er birth from 150 nmo// 100 mg/hr to tS nrnoV 100 mg/hr at 7 days of age: it increased thereafter to 170 nmol/ 100 mg/hr at 4 weeks of age and re- maioed at that level throughout the entire period studied. Glucose uptake and lactate production were highest io fetal lunp; fatty acid synthesis from glucose was alw highest at this age. 7bese observations show that late fetal lung has a high metabolic activity that oorrespoods to the high synthetic do- awnds of a rapidly growing and di/ftrestiatiod orpa. N.mo.h, M., Sbechter, Y. and Iluooab. P. Pediatric Research 12:93-100, 197fa. O/Arr .upp.rtr National Institutes of Healtb. From the Deputment of Physiology and Biophyaica, Georgetown University Medical School, Washingtoo,1) C. 33 12

r t I.IpOPROTF.IN LIPASE ACT7VITY AND BLOOD TRIGLYCERIDE LEVELS IN FETAL AND NEWBORN RATS Lipoprokie lipue u known to play a role lo clearing circulating W- slyceride, but it may aks.o have a special fusction In Ihe growth and matura- tioo of individual organs. To Investigate this, /ipoproteio lipse activity in lung and beart was studied in fetal (17-22 days of gestation) and newborn (day of birth until 30 days of age) rala. Blood triglyceride kveb were measured at all ages beginning at 17 days bo rle% and eaayme activity in epididymal, omenlal, and parametrial adipose titrua wr tested after 1!8 days of age. The developmental pattern of lipoprokis lipass diQered markedly in fung and heart. I.ipoprotein lipue activity wsr abeM In Ihe heart until Ilve hours be- fore birth; it increased gradually after birtL from 7.9% of adult activity to 17% at three days, 59% at eight days, and 73% at IS days. 1lr: activity dropped sharply from 75 to 49% of adult activity at weaning (day 21), and adult levels were reached at 24 days. 1s Ihe lung, eoatrasted to the heart, lipoprotein lipase activity wr high is the fetus (R4% of adult acti-ity), de- creased immediately sfter birth to 45% of adult activity at two days, aod remained at that level up b 11 days after birth. Enzyme aetivity rtarted to rise again at IS days and reached .duN levels at 21 days of age. Adipose tis- sue was present in trace amounts before tk+e age of two weeks. This qudy, which shows that lipoproteio lipase activity ia high in fetal lung d•iring the period of marked surfactant synthesis, suggests that circulating triulyceride- fally acids are used by the fetal lung for surfact.at synthesis. Naneouli. M. er at. ledlank Research 12:1112-11 J6, 19711. Other alpportt National Institute of Health. From the Department of Physiolop aod Biophysiu. Georgetown llnivenliy Medical School, Washington, D.C. QUANTI TATIVE CIIARACTERISflCS OF THE PEYRTER (APUD) CELIS OF TIIE NEONATAL RABBIT LUNG IN NORMOXIA AND CHRONIC HYPOXIA In this study of the quantitative characteristics of Feyrter or amino-pre- cursor-uplake-decarbosylalion (APUD) celb, counts were made of the APUD cells of 1-. S-, and 10day-old white New Zealand rabbits in normosic and chronic hypoaie states. A li6ht microscopy survey for nerve fibers in the epithelium was also carried out. Results showed that the ulls, both singly and in groups, were present in the epithelium of sd1 the intrapulmonary airways, although they were more frequeotly seen in the small bronchi and respiratory bronchioks. The number of Feyrier cells declined from one to 10 days of age (n normosie rabbits, whereas in the eaperimenul group (chronic hyposic ), the number of cells was low at one day of aee and no signifkanl changes occurred during the subsequent nine days. There was some indication of de- granuFslNiis of cells in the hyix+sic group It is ppnsibk that environmenlal snd/ar phyruilutarl factors asuxuted with the start of etlrautetine life, or 34 I lung devebpment,emay affett the apparent oumber aod probable level of aa tivity of these celb. These changes seemed 1o be enhanced by hypoaia, as observation which suggests again Uut these cells could poribly represent stor- age sites for 3-hydrosytryptsmioe. In this sludy, alw, intr.epithelial nerve flbcn in bronchi and bronchioks were found but they were -sol hmited to inner.ations of Feyrter cells or related eell bodies. Ilern.odez-Vasquez, A., W/d, l. A. and Quay; W. B. Thorax 32:449416, 1977. Other supports College of Agricultural and I3fe Scieooa, University of Wiscoasin, Madiaoa. From the Lkpartment of Veterinary Scknce, aod Neuroendocrine Soctb., Waismao ('entcr on Mental Retardation and Human Uevelopmeot, University of Wisconsio, Madison. A RADIOAU'TOORAPHIC STUDY OF THE NEUROEPITNELIAI. BODIES OF THE LUNGS IN FETAL AND NEONATAL RABBITS In a group of while New England rabbiu, 29-day-old fetuaes wer. I.- 1ed wbcutanewrly iw rfero with tritialed thymidine and the does were killed with sodium peatobazbNal injoetion, so as to obtain 2 fetuses for each o. the following stages: 2, 4. 6, !, 10, 12, 14, 16, 20, and 24 hn efler ail- thymidine injection. In a seoond group, I-day-old rabbita of the same breed as the fetuw, were also injected subcutaneously with sH-Ihymidine, and aacri/ked at 2, 4, 6, la, 10, 12, 14, 16. 16, 20, 22 aed 24 hn after injecton. Multiple samples Iaken from all lobes of Ihe lun6 were processed for p.raaln sectioning and radioauto6rarns were ezamined to establish whether the neuro- epi/helial bodies (NEBs) were undcrlioing mitosis and if so, to eaamine pos- sible variations in mitoMc rates and labeling indicea al the two different a/aloea studied. Results showed that thero was no uptake of Iritiated thymidine by 1he Nt!Bs and no mitotic fltura were found there at any of Ihe stages aludied. Also, the NEBa were not derived at this lime lrom prdiferations of other kinds of epithelial celb in the intrapultnonary, airways. These results stroatly suggest that the difference in numbers of NEBs previously observed by these authon, between the 29-day-old fetus and the I-day-old rabbit, is due either to regranulalion or acquisition of argyrophilie material by the NEBs or to diflerentiation of other epithelial lypes. It is eoncluded that Ihe NI?Bs am composed of welldilferentiated cells, which have a reduced capacity to un- derooo mitosis, during Ihe neonatal period. Hernandez-Vasquez, A., Wlll, l. .t, and Quay. W. B. Celi and Tissue Research 186:207-207, 1978. Ot6er support: College of Agricultural and Life Scienees, Univenity of Wisconsin, Madison. From the 1)epartment of Veterinary Science and Neurexndocrina Soction, Waisman ('cnter on Mental Retardation and Human Ikvelopment, l)nivenity of Wuconsrn, Madison. 35

I i i I fSi i i .~.~. I t9OM8ESIN-L1Kt31MMUNORt3ACT1V17Y IN TNB LUNG In this e:perimental study of the eadocrioe aspects of the lun6, human fetal and oeooatal hmp, ranging In age troeo eight weeka of gestation to tour osontha poanatal, were obtained froar 25 male and femrde wbjecta. Samples of tissue were taken from adjacent Inner, middle aod outer regions of each lung for Immunocytocbembtry (IC) and radioimmunoaaay (RIA), and aotisera directed to the C-tenaioal portion of amphibian bombesin and two bombesio analogues were raised io rabMb is several different ways. The optimal antiaerum for RIA waa foasd b be that lo pute amphibian bombesio and to the /Lys'ibornbeain aoalogte, w<tiereas for IC it was antiserum to the mr'ibombeaio analogue. Results •bowed cells with bombesin-like Immuno- reactivity nuiny in the bronchial and bronchiotar epithelium of fetal and neon•Ial lun6, both as single oel4 and aa kroup.. This immunoreactlvit7 was found by RIA to range from 2 to 43 ptnol per gm wet weight of ti•sue, whereas in preliminary atudies on human adult lung tso atls containing bombtain-like immunoreactivity were detected. This age-dependent d'astribMNion of cells containing bombesin-like Immunoreactivity, ssuggests that these eells may be of signiAcance during development and birth. tlombesio itself is one of a growing group of pepfidea found to be common to brain and Cut that seem to act both as neurotrammitten and aa local or ayslemie tarmooes. While the role of bombesin in the lung i• atitl specula/ive, it is imporunt Ihat, 40 years •lter Peyrter postulated an endocrine role for the lung, an endocrine peptide bas been found in the APUD cells of human fetal and ucoaatal lung. VYharton, l. er .l. ( W itl,1. A. ) Nature 273:769-770, 197t1. OtAer aappo rtr Medical Research Council, Cancer Research Campaign, and Stiftung VolkawaIcnwerk. Prom the Departments of HiNochesnitdrp and Medicine, Royal Poettraduate Medical Schtwl, Hammer.mith Hospital, Laodoa, Eagl•nd. IS i71E LUNG A PARA-ENDOCRINE ORGAN? lo addition to gas eschange, the lunp conduct activities that are sini- lar so those of endocrine glands. TAeir uo'tqtse ultraatrueture and pssitioo In the cireulator7 system enable Ihem to process certain hormones and pro- hormones (l.r.. Neraids, biogenie amine., proMaglandins, polypeptide hor- rnooes, and adenine nuckotidn). Conseqiuently, they probably Influence ape- ciAc actions of distant /issues and otgans, although they actually ryntheaise and release only prosta{landin related substances. While the intact lungs ae- ketively prooess only specific analogues within chemical groups of hormonea, this discrimination is lost when their structural integrity Is disrupted, indieat- in6 that in the natural state entymca are so partitioned among different cells and ore•nelka that some have accer to circulating hormones whereas others da mN StKh distlnction it a major determinant of the hotmone: fate and of the ulrsnutr d/.1M1\ItM.n ot the mtt.t..dic pr..fuct. Specifically dncus.ed art the htr. .Intmt d.cn pa...re thn-th the lunp of bradykinm, •(wdtnt htl..t.n..r .h..h ~% i.-t.tlt in..t..urd and of anrn.ttnsin 1. which is tn I activated through Its•conversioo to Ib lower btxnologue angiotenein II, a awat polent hypertensive. 'ibw, the ability of the tvnp to elimioate a 6oraaatr that lowen blood pressure while forming one 1lut raisea it, suggests a possible role in the regulation of blood preanure hoerowtasir. It also appears that a single ensyme, ankiotensio converting enzyme (kiaioaae 11), may aocouot tor both the inactivation of brad7kioio and the eoaversion of aopoleaain. In add/- tbn, it sow seema likely thot the htep uo also form aokioteaain 111, an a/do- aterone secretosogue that Is eveo nsore potent than angioteods H. It may be, however, that this particular mochaaiun in important for twaintaioios wuaioed elevation of •yuemk arterial blood prtnwrre levels only under wsch ciraws- danoes u result from trtatmenl with diuretia or streoutws e:erci.e, abat Y, when she plasma volume Is low or wheo the plasma reaio level i• eatraney high. 11 i• probable Ihat ahe /otal clarificalion of the rok of the htop io ther hoateo.talic mechanisny will be /ound at the cellular and molecular levels. The remainder of this review etamines Ihe variow sludia that have led to the formulation of /hcae concepts and justifiably ends by asking: la the hty a p.ra-eotlocrine organ? Ryan. 1. W. and Ryan. U. S. Tlie Amrrlcan lorrnd o/ 6lrJiclwr 67:S9S-60), 1977. Other wpprtr U. S. Public Health Servioe, Hartford Poundation, Inc. and the Heart Association of Pahn Seach County, Fla. From the t)epartment of Medicine, The New York Hoepitdl'ornell Medical Center, New York; the Papanicolaou Cancer Research lu.litute, and the De- partment of Medicine. University of Miami School of Medicine, Miami, Fta. ISOLATION AND CULTURE OF PULMONARY ARTERY ENDOTHeL1AL CELLS It is increasingly evident that endothefial oella are rmscb more than a mere mechanical barrier between blood and parenchyma. Since they are known also to diRer struclurally from one vaacular bed to aoother, it haa beeo suggested that they may d'ifer functionally as well. In order to further teat this hypothesis, however, endolhelium smW Aral be obtained from vari- ous aources and grown in pure, welleharackrized culturea. To facilitate such studies, the means of iaolation, culture and characterization of calf pulmonary artery endolhelium are detailed in this report. Theae cells may have siriA- eant importanoe in terms of specific metabolic aetiviries since there an inN- eation• that the lunp, and pulmonary endolhelia) cells in partieular, have a major influence on the mechani•m that regulates rhe hormonal composition of systemic arterial blood. Ryan. U. S. rr d. Tisst.a d Cell 10( )): 311-534, 1975. Other arpprtr U. S. Public Health Service and the /ohn A. Hartford ' Foundation From the Ikpartment of Medicine. Univer.ity of Miami School of Medicine. Miami, PIa. 37

SURFACE RNPLICAS OF PULMONARY ENDOTHELIAL Cti1.LS IN CULTURE Although numerous enzymes are known to esisl on the surface of pul- monary endolhelial cells, their topographical d'utribution is unknown. De- scribcd here is a simple method which uses an unmodified critical point drying apparatus and a high vacuum freczestch unit to prepare surface rcplicas of pulmonary endothelial cells. Thin lechsiqu, should be applicable to all eelis in monolayer culture. F-ndothelial eeW Srowe on cover slips or Slass slides were washed free of the culture medium, dted, dehydrated, and critical point dried. A 8alzer s freeze-etch unit wu used to prepare surface replicas. These have certain advantages over other types of preparations in Iha1 they can be examined with the resolving power of Ihe transmission electron microscope d and they can be used to esamine the aurfaw of whole cells. This technique may prove to be particularly useful for mapping specific enzyn.es, reccplors, and cell surface factors with the aid of markers attached either to specific antibodies or to subslances such as /ecliss that bind specifically to surface Rroups The ppearance of pulmonary endothelial cells in surface replicas is dc:cribed as a first step toward establishing a basis for such studies. Hart, M. A. and Ryan, U. S. TiUfue 6 ('.ll 1l)()):441-449, 1978. Other support: U. S. Public Health Service and the John A. Hartford Foundation, Inc. From the Department of Medicine. I/nivenily of Miami School of Medicine, Miami, Fla. SPECIFIC METAHOI IC ACTIVITIPS OF PULMONARY ENI)4)TTIELIAI. CELLS This brkf overview of lung metabolism focuses on interactions of the lung with circulating substances that ue processed actively by pulmonary endothelial cells. It is now knows that Lhe lunp are capable of sekclively processing a large number of subslasces of very different chemical types, in- eluding biogenic amines, adenine nucleotides, proslaglandins, polypeptides, drup, and lipids. Furthermore, pulmonary endothelium may contain enzymes and other agents active in coagulation and antiooagulation reactions. One enzyme of 1he pulmonary endothelial celis, angiotensintonverting enzyme, is capable of Inactivating bradykinin and of forming angiolensins 11 and 111. In addition to their specific metabolic ac/ivities. Ihese cells may also Iwsseu en- cymes Inhibitors such as .,-macrokarbulin. Although endolhelium of other vascular beds has similar or ldenlical properties, the pulmonary crKlothelium is uniquely silualed, between the central venous and systemic artrria/ circu- laliun., to affect specific functions of the lungs and specific functions of dis- lant organs. Thus, kinins and some pro.laglandins and biogenic amines do not survive passage through the lungs and do not enter systemic arteria( blood. In contrast. the quantities of angiotensins 11 and III which reach peripheral organs and glands via the systemic arterial circulation are probahly primarily dclcrmined by pulmonary endothelium. Rr'ln, 11 S and Ryan, I W. I In: S.nden, C. L. er d. (eds.): pulmowary ALrrooAase and E'ltheled CeQr (16th Ann. Hanford Biol. Symp.), National Technical Information Center, Energy Research A Devetoprnest Administration Seriea 43. U.S. Dept. of Commerce, Springfkld, Va., 1977, pp. 11S-14s0. Other awp'ort: U. S. Public Health Serviee, Hartford Foundation and the Heart Assocutan of Palm Beach Couoty; Fla. From the Papanicolaou Cancer Research Institute and Deparlment of Modicine. University of Miami School of Medieim, Miami, Fla. PULMONARY EtNI>ryfllF.l.IAL CELLS: KININASL311 AND OTIINR PL'PTIDB HYDROLASP. ENZYMES The present study was started /o (ocus attention on the fact that kinisrs 11 is not the only peplide invdved (n the processing of circulating bradykinis and angiotensin I by intacl lungs. While aniliolerria I aod bradykinin disappear during a single passage through the pulmonary eirculstion, these polypeptide Iwrmones are not lakea up by the lungs but are hydrolyzed by enzymes on or near the luminal surface of the pulmonary endothelial cells. Kininase 11 (aniliotensintonverting eszyme), a dipeptidyl earboaypeplidaae with tM capacity of inactivating bradykinin and of converting anRiotcnsis I to ila potent lower homoloRtrc, selliotensia 11, is described in detail here. Ilowever, the lungs contain other enzymes that sre capable of inactivating bradykisia and of metabolizing anpolensin I to yield products other than angiotensin 11. These also are likely to be situated on the lumissl surface of endothelial ulls. Two of the eszymes are capable of removing the N-terminal amino acid reaidues of bradykinin and angioteosin 1. For bradykinin, the removal of the N-terminal residue yields biologically inactive products. However, the removal of the aapartyl residue of ansiolessis I yidda a precursor of anRioleosM 11/. Interestingly esuubh, close observation of kininoe 11 shows that this enzyme eae aho convert des-Asp'-angioknsis I to angiolensin 111, a compound aves more active thas angiotessin 11 is stimulating the secretion of aldosteroae. la wmmary, this p.per. In adefNios to considering the apernative tates of theas polypeplides, attempts to n.ssess the selectivity of action of kiainaae 11. Thi, enzyme while acting as a dipeptidyl carboaypeptid..e can, in fad, hydrolyze a wide variety of polypeptide substrates. However, the relative aflinity of the enzyme for different substrates varies enorsnously, suggesting Iha/ this type of selective action may be of physiological importasoe. Ryan,1. W., Rym, U. S. sod Chiu, A. T. Is: Sanden, C. 1.. et a/. (eds ): rrlnwnory M.rropbage, .nd EOlr11e7Jaf Crlb (16th Ann. Hanford Ilial. Symp.)t National Technical Information C<oter, Energy Research & Developrnent Administration Series 4), U. S. Dept. of C.ommeree, Spriagfield, Va., 1977, pp. 141-14e. OtRsr arp'orrr U. S. Public Health Servioe, Ilarttord Foundation and the Heart Association of Palm Beach County, Fla. F om the P,panicolaou Cancer Research Institute; Ikpartment of Medkine. University of Miami School of Medkine, Miami, Fla ; and the Medical ('olkge of Virginia, Richmond. 38 39

11UMORAL CONTROL OF ARTERIAL BLOOD PRESSURE: A ROI.@ FOR TH8 LUNG? White the primary function of the hmp Is the eschante of tua. between blood aod dr, the past 10 yean have nhorre that the lungs can sekclively process a wide range of hortnonea, prohormooes, and other escitatory asents that have direct or Indirect eRects on the trystank arterial blood pressure. It is known now, for instance, bow bradylinie and antiolensin I are rm:tabolized at the cellular, subcellular, and molecvlar Ieveh. More speciAcalty, recent studies have defined in detail bow a.w Mmg enzyme, antiotemin-converting enzyme, can participate in the diwieatb" of a hypoknsive hormore, brady- tinin, while forming antiotensin 11, oea of the most polent hypertensone agents known. Tris enzyme Is found along the hu+inat surface of the endothelid cells lining both the larte pulnsonary vessels and Ihs capillaries. As blood passes through the pulmonary vascular bed, aegioteosin<onverting enzyme is washed with its owe aubslrates ia a cwntirNwrrly eowioR liquid phase, and the reaction products are released direcdy Into tlt" systemic arterial clrculation. The enzyme is found in vascular beda other due that of the lunp, but only the lungs export the reaction product, angjolerssin 11, to the systemic ecrculation. Also, the lungs cannot detrade angioleruie 11, which ie.ures its avail.biliry for export. These features suggest a role for aediotensin<oevertint enzyme in blood pressure homeostasis, but the detailed rehtiorohips of the luop to humoral control of the arterial blood prsrurc remain to be defloed. Ryae, /. W. and Ryew, U. S. C.rdlovarcrfa. Aledk/ne 7:S)1-3)7 rr pactbn, 1971. Other st.'p~rt: U. S. Public Ilcalth Service, John A. Hartford Foundatioo, American Ileart Association, and Ileart Association of Palm Beach County. Fla. From the Department of Modkine. University of Miami School of Medicine, Miami, Fla. NEW SUBSTRATES FOR TIIE RADIOASSAY OF ANGIOTENSIN CONVERTING ENZYME OF ENDOTHF.LIAL CELLS IN CULTURE Three simple acylated tripeptidea were synthesized as substrates for angiotensin converting enzyme in order to provide the meaos to measure Its activity in cultured endothelial ee0s. Specifically, these preparations were p-I'Hlhenroyl-Pro-Phe-Art-OI( (11), p-1•H/benzoyl-Phe-Ala-Pro-OII (1) and p-I'Hlbennoyl-0Iy-His-Ltu-OH (111). Substrate I haa the lowest Km (12.5 rM) and is the most sensitive for assay. The angiolensie converting enzyme content of 10' cells can be measured afler a)0-minute Incubation period al )7' C. TLe rad'aactive reaction product (s separated from the substrate by estraction of the acidified reaction misture with an organic solvent. Tlrc rate of product formation is then quantified by liquid scintillation counting of the organic phase. Substrate 111 can also be used for this aaay, but requires longer in- cubalion OJ MNns/ and high uit concentrations (0.75 M Na,SO.). Sub- suate 11 is rwrt y.ec rfic and rs hydrolyzed by more than one aufothelial cell enrymc Ryan. J. W., Chung, A., Martio, L C., and Ryan. U. S. Tiuue d Cell 10( )):335-362, 1971. From the Department of Medicine. University of Miami Schoo( of Medkin., Miami, Fla. A SIMPLE RADIOASSAY FOR ANOIOTENSIN-CONVERTINO ENZYME This paper describea a radio.s..y method in which p-/stllbenzoy/gly- cylRlycyltlycine ('11-labeled hippuMtlycyltlycine) is used as substrate for aegiolensiotonvertind enzyme. Since enzyme activity is measured in lerms of the rate of release of 'H-labcled hippurate, prior di.lysis of serum sueplea and Iiaue homoteeatn is not rcquired. TUe product in separable from the wrbsurate by eatrauion of acidified Kaction mislurea with ethyl acetate. Tbis way was used to measure angiotensin<onverting enzyme activity In sera of mae, rat and guinea pig, and in homogenates of rat kidney and guinea pig bn=. To further test the specificity of the assay, serum samples (from healthy vdunteen, p.tients with active aareoidosis and guinea pip) were examined for their ability to form radioactive products other than hippursic. In each esperiment, all radioactivity was associated with hippurate and unhydrolysed substrate. The major advantages of this assay over those used previously are simplicity and rapidity. For eaneple assay results for human serum anSio- leesiatonvertieR enzyme can be obtained withi. 1.5 hour. of receipt of serum samples. Within the limits /ested here, the assay appears to be specific. llowever, interference by hitherto unrecognized enzymes of abnormal sera must be ruled out. I Ryaa, ). W. et J. (Ryae, U. S.) dloc/kndrd lournal 167:S01-SW, 1977. From tbe Department of Medkine. University of Miami School of Medkin., Minol, Fla. 111. Ne.rt .nd CJrcul.Hon PURIFICATION AND CIIARACTP.RITATTON OF 1.ECfININ:CIfO1.P.STP.RO1. ACYI.TRANSFERASP. The purification of kcithin:chokaterol acyllraederase has previously been reported. The newer method described here, however, yields a preparalion purified 16,000-fold and free of high density lipoprotein and albumin. In coo- trast with the product previously obtained in this laboratory, which was only hdf as pure and contained two maior proteins, the one obtained by this newer technique showed on/y one band upon polyacrylamide tc) electrophoresis. SDS- uree polyacrylamale tel electrophoresis and ialKlectr/c f.x:using, and one arc in immuoodiRusion against a toat anliserum preparatioo. The enzyme was 40 41

determined to be a tlycoprolein with a molecular weight of about 70,D00 and a pl of ).7-1.0. Oustow, E., Varma, K. Q, and Soloo, L. A. Scandlnovian lorrnd o/ Clinkd 1 LaAorarory Investigation )6(Sltppl 150) :1-S, 1978. OtAer.rpportr U.S. Public Health Service. From the 11pid Rexarcb Laboratory. Department of Medicine, Temple Uoi- versity Ilealth Scknces Center, Philadelpbh. RPLATIONSIIIP BITiWIiPN 111011 DENSITY LIPOPROTEINS AND 111P. RAl EOP fN VITRO SERUM CNOLESTEROL ESfI?RIFICATION Mounting evidence suggests that the Incidence of coronary heart disease is inversely relaled to the concentration of high densily lipoprolein cholesterol (IIDLl'). Also, the plasma concentration of HDL-C is foveraely related to the body pool of cholesterol suggesting that HDL protects against the development of coronary heart disease by transporting tissue cholesterol to other ortane (liver) for escretion. In addition, HDL has been shown to be the substrate of choice for kcilhinchokslerol acyllransferase (LCAT), the enzyme responsible for the esterilkation of human plasma cholesterol. This study of geriatric aub- tects attempts to determine any relationships between LCAT. NDL-C and the rate of esterifkation which might indicate /hat the enzyme i activity is one of the factors involved in IIDL's supposed antialherosclerotie effect. The rate of cholesterol eslerifkalioo and the IIDL-C concentrations were measured in vitro in serum samples from nine men and 18 women whose age ranged from 74 to 95. 7he resulls clearly show Ihat asymptomalic geriatric subjects can have lower HD1.C levels than those considered normal in persons under 60. Un- especledly, however, the biochemical data also reveal that the cholesterol esteri(kation reaction rate is more rapid In sera containing ku than 40 mt/d) Nl)1.-C than In samples with more than 40 mt/dl. No correlation appeared between cholesterol eslerifkation and a proleia(s) which can produce an immune tamma globulin Ihat completely inhibits in vitro serum cholesterol e.terilkalion. These rewlts are discussed with reference to the presumed re- lationships between HDI.C and atherosclerosis. So1o0, L. A. sod Varma, K. O. Standlnavian Journal of Cl/nkd • Laboratory Fnvenltarion 38(Suppl. ISO): 72•76, 1978. OtAer support: t/ S. Public Health Service. From Temple lloiversity Ilospital and School of Medicine. Philadelphia. IMMUNI)t (N.1('AI 1•VAI IlA t U)N t)P 1CA 1' 1)1'Fll'IFN('Y Ibn saN1r aurmpu Io rv.lu.lt the nsture uf Ihc 1('A1 dcftciency en• c+.umcit.1 m i..• s.,b/c.is hy intan1 nf an immun,kIJlusion ud tmuronumhi bition technique, utilj;int two different antibodiea obtaine;l from two diRereat toats to test a highly purifled LCAT preparation against LCAT-0eOcknt sera Tbe reaction of identily between normal aerum, defkient sera and purified LCAT suggested the presence of a catalytically inactive en:ytne in the da- lkient sera. In normal serum, excas antibody inhibi/ed 50% of the LCAT activity. One of the antibodies faikd to inhibit the enzyme when the Wt system contained escess deficient serum, seemingly supporting the above eon- cluiwn. The other antibody preparation, however, inhibited LCAT activity even in antigen escess. The precipitin lines observed in MnmunodiRwiqo do oot seem to represent serum LCAT. In view, of the higher 1i1er of antibody observed In immunoinhibilion esperimenta with this antibody, a question eow arises as to whether immunoinhibiuon by the antibody will also be abdiahed at lower ralios of antibody to defkienl serum. , Varma, K. (1., SofoQ, 1.. A. and Frohlkh, 1. Scandlnavian Jorrrnal of Cqnkal d Laboratory Investigation )ti(Suppl. 1S0): 6-11, 1978. From the Lipid Research Laboralories, Department of Medicine. Temple Uni- versity. Philadelphia; sod the Dep.rtment of Palholoty. University of British Columbia, Vancouver, Canada. a TNE INHIBITORY EFFECT OP 7-KETOCHOLESTEROL ON Tt1E CHOLESTEROL UPTAKE BY THE ARTERIAL WALL Previous esperimenls have ahown that 7-ketoehoksterol inhibits the up- take of cholesterol by the artery in .itro, but In vlvo Inhibition proved difikvlt to demonstrate. This report eaamina the mechanisms underlying this in- hibilory action on cholesterol uptake by the arterial wall. The sorus of hepaleclami:ed rabbits injected with sohrbilized 7-ketochokuerol failed to show In vivo inhibition of choksterol uptake. Neither wu choksterol uptake inhibited when rabbit carotid arteries were perfused /n vitro with pla.mm pooled fronm hepaleclomizcd animals injecled with solubilited 7-ketochokslerol. llowever, porcine coronary arteries demonstrated dgnifkant cholesterol uptake inhibition when they were perfused wilh iaologoua plasma containing par- ticulate 7-kelochoksterol. These resulta demonNrale that only the p.rticulate fraction of 7-ketochoksterol inhibits cholesterol uptake by the arterial wall. llrus. the possibility that the inhibition may not occur at the arterial wall but results from a physical Inleraclion between particulate 7-kelochoksterol sod e/wkslerul in the perfusale must be considered. Sarma, 1. S. M. and elng, R, l. Journal of Motecular and Cellular Cardiology 10:197-201, 1979. Other support: lhe Hoover Foundation. • From the Iluntintlon Memorial Ilospilal and lluntinti.•. Insthute of Applial Medical Reseaich. ('alilmnia lnstilute of 'Iechnoluty, 1'asadcns, and the University of Suulhern ('ablornia, I os Anteks. 42 43

1 l3FFHCT OF H1GH DENSITY LIPOPROTFJNS ON THE CHOI.PSI'(iROL UPf AKN BY ISOI.ATED PIO CORONARY ARTERIES In this series of eight esperimenu dealiog with the effect of added high density lipoproteins (11D1.) on chokaterol uplake, pig coronary arteries were perfused In vitro with isologous plasma containing sll-I,2-choksterol with and without added 11D1. from the sarne species, and the cholesterol up- take by the arteries was measured. Raulta showed that the arteries perfused in the presence of added HOL took up sijaificantly kss cholesterol than did the controls. Cholesterol uptake by Ihe cootrol group of arteries averaged 180 :t 34 n nwles/g, whereas tbe arteries pertused in the presence of added Ill)1. took up only an average of Ilt :t 2 a rnoks/l. The difference between the two values was significant (p<0.001). While one possible reason for this observaNon may be that HDL direcdy inter(eres with the entry of chokstero) intn the arterial t'ssue, a second possibility is that lID1. is able to promote cmua of cholesterol from the tissue. There is some supporting evidence for thia, and from this standpoint the results presented here, which co.nplement epiJen,iological and cell culture uudies by olhen, seem to flt into the general hyprwhesis that HDIL promotes efllux of cholesterol from tissue. Sarma, 1. S. M.. Tschurtschenthakr, O. V. and Iint. R. l. Artery 4()):214-22), 1978. OOther support: Hoover Foundatioo. Prom the Uoiversity of Southern California School of Medkine, 1d Angeles; Huntington Memorial llospital, California Institute of Technology and the Huntington Institute for Applied Medical Research. Pasadena. ('OMPARATTVEi E?FFE('iS UP ('HRONI(' FTIIANO1. AND AC@TALUIiHYDII FXPl)SURP ON MY(X'ARUTAL FUNCIION IN RATS In order to develop an esperimental model that would permit separate monitoring of the effects of ethanol and acelddehyde (one of its mclabolita) on cardiac performance. two metabolic inhibilors, 4-methylpyrazok (4-MP) and pargyline (PAR). were used to iodepeodently raise the blood levels of ethanol and acetaldehyde respectively. Rats who received as much as 36% of their total daily caloric supply (rom ethanol for a period o( three to (our weeks were sacrificed after injection wilh eilher saline or one of the in- hibiton. Blood samples were measured for their ethanol and acclaldehyde kvels. Tbe milochondrial respiration. /n vJtro contractility of Rlycuinated heart muscle fibers and myocardial protein synthesis were determined. Accord- ing to the results, either 4-MP or PAR administered in addition to ethanol damaged mitochrondrial respiration and myocardial protein synthesis more severely than did akohol ingestion alone. These data illustrate that both acetaWehyde and ethanol are capable of damaging the myocardium. and that high concentrations of either substance can severely aflecl its metabolism. Wei.haar, R, Bcrruglia, S, Ashikawa, K.. Sarma, 1. S M., and 8ins. R. I. 1'hr lourn,d o/C'Lni.ulPharnru,.rloRy 1R(R39):177 )R7, 1978. 44 Other support: U. S. Public Health Service and the Hoover Foundation. From the tluolioglon Institute of Applied Medical Research and Iluetiogloa Memorial Hoapital, Pnadena; the University of Southern Cali(oroia, Los Angeles; and the California lmtitute of Technology. Pasadena. CARDIAC INDEX AND INCIDENCE OF HEART FAILURB CBILS Because there appears to be a relationship betweea pulmonary blood (bw and the presence of iron-ladea macrophapn is bronchial washiop, the percentage of these ulla was assessed in the apwurn of heart disease paticou before their cardiac index was measured. An inverse relationship was found between the percentage of these qcatkd heart failure cells ia aputura amcan and the cardiac indes of the palienla, teprdkr of the (ype of cardiac illness. The highest proprxlion (73%) of theae cells was found in association with the lowest cardiac indes, or I Ii1er/min/aq m of body wr(aee area (BSA). In eoa- trast, when the cardiac indes was around 3 liter/miNaq rrs BSA, less than 2% of the macrophaRes contained iron, which is consistent with previous obaervr /ions in healthy controls. However, widely scattered and seemingly uordaled values were obtained at intermediate cardiac index kvels, presumably reflecting individual variations in the degree of activity of the aubjects' reliculoendothelial system. I Friedmao-Mor, Z., Cha(on, l., Turodor(, H. and Orkin. L R. Archfrei oJ Pathology .nd lALporatory 1/edlclnne 102:41t-119, 1978. From the Departmenu of Anesthaiology, Albert Einstein College of Mediciec, The Brona, and New York Univenity Medical Center, New York. TRACHEOBRONCIIIAL CYT'OLOOIC CIIANGES AND ABNORMAL SERUM HEME PIGMENTS IN HEMORRIIA(7IC SHOCK Among the pathophyaiologie and metabolie aberrations caused by hna- onhajic ahock are lysis of erylhrocyles and degradation of free serum bemo- Rlobin into abnormal heme piRmenls which are believed to be toxic. /1 had previously been shown that significantly elevated numbers of iron-laden his- tiocyta, as demonstrated by the Prussian-blue staining melhod, appear ia the tracheobronchial secrelions of patients in hemorrhagic shock and, during cardiopulmonary bypass, in those undergoing open-heart surgery. Because the hisliocyles may ingest abnormal heeneyiRments as part of a defense mechanism, this study altcmpu to correlate the percentage of iron-laden ulls in Ihe tracheobronchial sccretions of patients in hemorrhagic shock with the presence of hemoglobin degradalion products in their serum, as delermined by acanning spectrophotometry. In general, there were fewer iron-laden hisliocytq when hemoglobin de1gradatinn was advanced Ihan either In the absence of abKormal heme pigrncnls or when Ihere were only minor degrees of dcRradatiun. Ihcse observations demonstrate the presence of abnormal heme pigmcols in the act-um 45

I I of humans in hemorrhagic shock, and suggest that few of the degradation products of hemoglobin remain in circulation when the degree of reticuloen- dothelial response is high. lAus, the dala preaeoted here probably reflect the reticuloendothelial prouss that eliminatea toxic products accumulated in the circulation during low-flow sutea. It is also pouible, however, that the resulta actually represent the liberation of myoglobin or of enzymes, such as cyto- chromes, perosidase, catalase and othen, that incorporate heme molecules into the circulation. Friedman-Mot, 7.., CAalon, I., 7Lrsdorf, H., and Orkin, l.. R. The Iournal of Trauma 17(11):/29-i31, 1977. From the Departments of Anesthesiology of New York University Medical ('enter, New York, and of the Albert Plmteio College of Medicine, 'Ihe Bronx. N. Y. ABNORMAL Ifl?MP.PROTEIN PATTERNS IN HEMORRIIAGIC SI(OCK Abnormal heme pigments have been described earlier in the aera of animats bled to hemorrhagic shock Ifere, electrophoretie studies have been run on the sera of patients in hemorrhagic shock in an attempt to identify abnormal heme pigments and to relate the appearance of some of these compounds to rnor- tality rates. Subjects for this study included 10 consenting adults who were suAerinR from hemorrhagic shock. In addition, blood umpka were obtained from: (1) five hcalthy volunteers, (2) three 21-day-old specimens of bank blood, and (3) two healthy patienu anesthetiud but not in shock. Results showed that the accumulalion of inetabolilea, caused by the impedcd eircula- tiun, degraded free hemoglobin into herne pigmeota, and their concen- Iralion then reached a level that esceeded the normal henre<arryinR capacity of aerum prolelna. All patienu In shock bad distinct bands e/ varying iotensity in the benzidine-stained strip. Nina patients had haptoglohin-hemo- globin bands, live patients also had a hemopeain-heme band, and four patients had an additional melhemdbumin band. 'il+ere were no deaths associated with the presence of haptoSlobin-hemo6lobia aloee In serum. However, as shock deepened and mortality roae, hemope.in-heme and mcthemalbumin appeared. The highest mortality rate (4 out of 3 cases) was found when bcah herno- pe.in-heme and methemalbumin were present. 1t seems, therefore, that ad- minntration of additional serum proteins to increase Ihe binding capacity of the blood might reduce the toxic effect of heme. Fricdman-Mot, Z., CAafon. I., Oorstein, P., Tbrndorf, It., ('huba, 1. V., and Orkin, L. R: TAe lor.rnal o/ Trauma 1s(2):104-107, 1978. Wnm the Ikpartmtnts of Anesthesioluty and Pathnlaaty. New York llnivenity f.lydu.l ('tater, New Yurt, and 1he 1)epartment of Anesthesiology. Albert I.n.tr~n I~..Ilete a•l MrJia ~nt 1 ht Ilr.m• N V 46 ROLE OF N1OH-f(IOI-ECUI.AR-WEIOHT KININOGFN IN SURFACP,-BINDING AND ACTIVATION OF COAGULATION FACTOR XI AND PREKALLIKREIN It is known that high-moletular-weight (M,) kininolien is a cofactor for the activation of Factor XI, prekallikrein and HaReman facwr, but several hypothescs exist in explanation of Ihis functional role. This study demonatrata that one mechanian by which high M, kininoRen acts as a cofactor for the activation of both Factor XI and prekallikrein in plasma is to bind both these molecules to the aurface where they are activated by surface-bound activated Hageman /actor. Experimenlally, when rs'1-Fac/or XI and r'al-prc- kallikrein were added 1o kaolin-activated nornril human plasma and plasmaa deficient in high M, kininoRen ad IlaReman faetor, reaulta showed that high M, kininotien was essential tor normal binding and cleavage of both Factor XI and prekallikreia on the kaoli• .urface, whereas Ilalleman factor was eucntid tor the cleavage but not for the binding. In normal plaama, tiA% of the activated Factor XI remained surface-bound, but 80% of the kallikreie did not. These findings are consistent with the hypothesis that. In the initial phase ol contact activation, high M, kininogen links both Factor XI and prekallikrein to the esposed surface where they are activated by wr/ace-bound activated Hageman factor. Once aetivated, the Factor XI molecules remain localized at the aite of activation,-in contrast to the kallikrein mokcuks that are found largely in the surrounding plasma. Wiuin., R. C., Bouma, l!. N., CocAraose. C. Q., and Oriflin, 1. H. proceedings of the National Academy of Sciences of the Unlted Srates of Arneriea 74(10) :16)6-4610, 1977. Ot/Ler .r pprt t National Instituta of Health. From the Department of Immunopathobp, Scripps Clinic and Reaearci Foundation, La 1o11a, Cal. SURFACE AND FLUID PHASE ACTIVITIES OF TWO FORMS OP A(?IVATED IIAOEMAN FACTOR PRODUCED DURIN(7 CONTACT ACTIVATION OF PLASMA This report describes the ability of the two forms of activated Harsna, factor (HF,) produced during contact activation of plasma to activate prf kailikrein and lactor XI. The infhrence of the wrf.ce on the cleavage of lactor XI and prekallikrein, and the distribution of these proteins between the sur- face and fluid phase compartments before and after their interaction with HF, were the particular fackm considered. In these e.periroenta, the 110,000 mokeular weight Iwo<hain enzyme'Ihat remains bound to Ihe sur/ace, termed r1IF„ was capable of cleaving surface-bound prekallikrein and /actor XI. Factor X/ remained bound to the wrfloce, while prekallikrein and kallikrein rapidly entered the supernate. A 2e,000 molecular wei6ht single-chain mok- eule released fu,m the surface dutinR contact activation, termed 0 IfP„ split prekallikrein. Inrt nnt factor X1. 'lhis reaction occurred whlthcr the substrate was surface brwrnd or in solution. lhcse data suggest that Intrinsic coagulation resulting (rons the activation of (actor Xt is  localized event thal 47

occurs at the site of contact activation and is caused by the action of a-HF,. On the other hand, kinin generalioo and flbrinolysis resulting from the forma- tion of kallikrein can be initiated either by r1117. at the contact activation site or by fi-HFs throughout the plasma. Theae two activities are kuaranteed further dissemination by kallikrein i rapid dissociation from the surface. Revak, S. D., Cochrane, C. G.. Bouma, B. N., and Orifflo, l. H. The lourrwi of Ei perimenrd Altdicinr 147:719-729. 1978. Other auprortr U. S. Public lie.h6 Service. From the tkpartment of Immunopatho{og,y, Scripps Clink and Research Foundation. I.a lolla, C.I. 1111? INTI?RA(T1ON OF rs-I-PRO'iE1NASt31NHIBITOR WI111 StrRINE PROi'EINASBS About 10% by weight of the proteins ie human plasma are known to io- bibit proteolytic enzymes. 7lrese proteim may have either a very narrow range of inhibitory specificity or very little specificity. While the major plasma pro/einax inhibitor. .-I-proteinase inhibilor (e-1-PI), previously known as .-1-antitryp.in, has a broad range of speeificity, it cannot inactivate certain enzymn. llws, in spite of the very high .-1-P) concentration in plasma. It seems certain that ..2-macroglobulin is more significant in the regulation of prnlcolysis by all types of enzymes of this clasa. 1lre potential role of ~t-PI in the regulation ol the coagulation process hu been a controvcnial subject for many years. If it were found to inhibit thrombin, it might have some significance In the regulation of thrombin activity in vivo. Newer procedures have made it possible to isolate .-1-PI from antithromlwn III activity and have allowed several conclusions to be drawn. Although the tole of this Inhibitor in eoagulation is probably minimal, based on its high concentration In plasma, its concentration in the acute phase state aod its ability to inactivate thrombin, a supporting role in Ihis process cannot be entirely dismisxd. 'ilre development of powerful synthetic peptide iahNtors provides new methods for studying the inhibitory activity of pluma proteiaues. Trerls, !.. Matheaon, N. and lohaaon. D. In: l.uodblad, R. L., Penlon, 1. W., 11. and Mann. K. 0. (ods.): CJrrmtnry .nd eiolory of ThromAin, Ann Arbor, Michigan: Ann Arlwr Science Pub- tlshers, 1977, pp. 431-440. Ot6er support: Natiooat Institutes of Hedth. From the Department of Biochemistry. TUe Univcnity of ()corgia, Athens. CONTRIBITTION OF CARI)IOP111.MONARY BARORF.('EPTORti ' TO '1111'. ('ON IRI )I. O1' 77I1i K 11)NI!Y The electrophysinlogic behavior of many of the eardiopulrnonary baro- receplnrs /us bccn eslen.ively e.ammed and a sreat deal is knr.wn abrwt 48 I the determinants of; their discharge frequency. However, their rok In the orpnism is not aa well understood. Whik evidence indicatea tbat receptors in the cardiopulmonary region ean, under certain eircumatances, eaert a pro- found influence on renal sympalhetic nerve activity, renal vascular resistance and renin rekase, it is difficult to prove etperimeotally that these receptors .r. Involved in the cohtrol of blood volume. Hrrwever, the available evidence in- dicatea that cardiopulmonary receptors with vagal aRerenta eaert a took inhibilion on both renal nerve activity and on renin release. The magnitude o( this inhibition appears directly related to changes in blood volume. Atriat r well as ventricular receptors can infhacnce the aecretioa of renin. Cardio- pulmonary receptors with vagal aRereota may also reAealy modulate renal prostaglaodia secretiorr. There is preliminary evidence suggesting that cardio- pulmonary receptors with .ympathelie aRerenta can inAuence renal nerve acGvity. le this paper, the limitations of previous studiea are outlined and a direction for future studies is suggested. PiaaRy, it is tooch.ded tbat alteratioe. in cardiopulmonary vapl aAerenl input and tbe resulting chantea in renal nerve activity and ie reain release are appropriate tot Iha maintenance of blood volume bomeo.tasi.. TArner, A(. D. Pe4radon rroceedi'nngs )7(S):1209-1217, 1978. Other support: National ImtitWa of Health. Prom the Department of Physiobsy and Biopbraica, Mayo Clink and Mayo Foundation, Rochester, Mim. IV. Neuroph.rntatology and Pky.iotosy DISCRIMINATIVE STIMUI.US PROPERTIBS OP NIaOT1NE AND NICOTINE-RELATED COMPOUNDS Nicotine, one of the drup moat widely used by man, has been observed to have very apecifk and sensitive discriminative stimulus (DS) properties 'The DS approach, which has been very useful in studying the mechanism of action of nicotine, measures the subjects' response, oot to a specific effect of the drug on behavior, but to the presence of the drug within the CNS. In thV Investigation, a series of eaperimenta was Initiated which studied the See- eraliralkm of the nicotine ()S to chemical analogues of nicoline arwl to various eompounds purporaed to harAe similar behavinral effecta. Usinl the DS behavioral paradigm, rats were trained to discriminate between the US eflecta of nicotine (200 y.g/kg, s.c.) aod saline using a two-kver, food-reinforced operant procedure (VI 15 sec.). Teo diQerent compounds were evaluated in rats trained to discriminate nicotine over a wide ranje of doses and .cron both subcutanern.s and intUavenlrieular routes of administration Oidy on. w.npound. )-pyridytmdhylpyrrolwline, produced significant nicotinclike ac- tivity and, although the experimental animals perceived this compound u it 49

they were given nicotine, it possessed only one-seventh the potency of nicotine. The niccNine stimulus did not generalize to compounds such as lobeline, colinine, or d amphctamine. One of the most striking results of these studies was the specifkity of the nicotine DS. indicating that Ihis paradigm mav provide a unique approach to studying the structure-aclivity relationships oC the be- havioral eflecls of nicoline. Rozt.-rans. l. A. er d. In: Biittig, K. (ed ): Sellaviora/ EQects o/ Nfcodne, B..el: Karger, 9979. pp. 70-82. From the Department of PhannacoloRy, Medical College of Virginia, Richmond. A('UMI'ARISON OF NI('OTINB AND STRUC7URAI.LY RCI.ATI'.f) COMPUUNI)S AS UISCRIMINATIVE S71MUU lhe discriminative cue produced by oiootine Injeetion seems to depend upon the central rather than the peripheral effect of this agent, and recent aludres have suggested that Ihis centrally produced cue occurs at N<holineraie receptors hut not at muscarinic (M) reuplors. Thua, relatively specific stim- ulus cflecls are involved Seven compounds were eaamined for generalization to the nimulus effects of nicotine, in order to investigate whether structural similarity elreru an analogous central cue and to elucidate the chemical con- flguratron crucial to its pnxhrctnm Of the compounds lesled, only )-pyridyl- mc/hylpyrndidine ( 1-PMI') generalized to the stimulus effect of nicotine. Pquivaknt nkoline-like responscs were obtained with tlx) pg/kg. a dose ap- prosimately four limes that used for the original nicotine discrimination. Tlre IiD„ for )-PMP was atxwt five times that of nicotine. None of the carn- pounds signifkantly blocked the nicotine cue. Nowever, the nicotine-like cue produced by 800 pg of 3-PMP was effectively blocked by meeamylamine but not by heaamethonium or by alrupine. llrerefore, 3-PMP appears to cause generalization to the nicotine cue by acting on central nicotinic cholino receplors, as was previously reported for the nieotine discriminative stimulus. The I:D„ for the blocking action of ineesmylamine on the stimulus effects of 3-PMP (s00 pg/kg) and of nicotine (200 M6/kg) was 0.32 and 0.20 mg/kIl, respec/ively. The use of this discrimination technique for comparing structurally related compounds is an effective as well as speeifSe technique. Chance, W. T.. Kallman, M. F)., Roseccauu, /. A., and Spencer. R. M. Drkish lorrnaf o/ Phannaroloty 6):609 616, 1978. From the Department of Pharmacolop, Medical College of Virginia. Rich- mond. A DESCRIPI-1t7N OF' TIIP. NI('OTINP, ST7MULl1S ANI) l I:STS OI' fIS (ll?NIiRAI /IA fl()N 10 AMF'IIHa AMINN. ihis aeries of c.pe+imenii was designed to investigate the discriroinalive slirmdus prrqemes uf nwulrne and ro detcrmine the degree of kencraluatton 50 of he n'r.otine stime)lus effects to those of d-amphetamioe. In each of three separate experiments, the subjoct's performance was assessed wioR a twokver operant procedure with liquid food reinforcement. In the nrst study, rats wero trained to discriminale between various dosea of nicotine (100, 200, or 400 pVkt) and saline under a VI-(S a schedule of reiniorcement. The second study investigated the time-0urat'an parameter of the nicotine atimulus by mooiloring discrimination between 40 pg/kg of nicotiae and saline under three different schedules of reinforcement. Oeneraliralioo of the nicotine stim- ulus (400 pg/kg) to the stimulue effects of several daes of d-amphctamioa (60, 120, 210, 480. and 720 pg/kg) was investiRalod is the third study. Re- wlts of /he dose-gencralisdion and Gmeduratios studies showed that the sensitivity of the rats to the nicotine cue wr directly related to the training dose under the VI-IS s scheduk. Allhoujh response ratta differed across the scheduka of reinforcement, the rat: sensitivity to the stimulus eflects of nico- tine was not aflected. l ack of complete Renerafaation of the nko/ine stimulus to d-amphetaminc indicated that the rals did not perceive those two drup as the same. In addition to these e.perimenlal findinp, this paper presenla sev- eral techniques useful for proper evaluation of the stimulus properties of Iea1 compounds. Chance, W. T., Murtin, D., Krynock, O. M., and Rorecranu, l. A. psycAophnmardosr SS:19-26, 1977. From the Department of Pharmacology, Medical College of Virginia, Rich- mood. I CONVERSION BETWEEN CONFIGURATIONAL STATES OP THt3 MUSCARINIC RECEPTOR IN RAT BRAIN AccordinR to these eaperimenh, reductive alkylalion of neural mem- branes by N-ethyl makimide (NEM) converts muscarinic acetylcholine ro- eeplors from a stale of low Io one of high aAinity for receptor allonists. This treatment does not affect the inleractions of mu.carinie antagonists with Ihe receptor. The NEM treatment also increases the proportion of rat tekncephaloa high aRonisl aA'inily recepoa from 34.2 to 53.4% of the total receptor popu- lalion; and it decreases the potential esrb.mykholine inhibition d)-quinu- clidinyl benzilale binding from a K, of 1.2 a 10-• to 6.9 a 10- •. These re- sults seem to support  previously proposed theoretical rnodel, which suggests the e.islence of two populalions of receplon with different aR~nities for agoo- ists. In lhis model, the anlagonisls supposedly bind to either receptor with a uniformly high a0inity, and the binding of a6onista and sulagr.niats is com- pelilive arwf mutually esclusive. , Aronatam, R. S., Hosa, W. and Abood. f.. G. European lournaf o/ PAarmocology 16:279-2l2, 1977. Other support: U. S. Public Nealth Seryice. ~ From the Center (ur Brain Research, University of Rochesler Scfwr,t of Mcdi- eino aod Dentistry. Rochester. N. Y. SI

INFLUGNCE OP SUI-FIIYDRYL REAGENTS AND IIP.AVY METAIS ON 111E ft1NCTIONAI. STATE OP TNE MUSCARINIC ACE(1f1.CHOLINE RECEPTOR IN RAT BRAIN Rat brain membranes contain several mobcular groups, )ocatei both within and contiguous to the receptor bindinR aite, that react with wl!hydryl reaaenls to influence muaearinic aeetykholine receptor binding. Oa! com- pound, pahbromercuribensoate (PCMB), reacla with a group(s) wi hie or under the allosteric control of the seeeptor binding aite to inhibit both atonist and antagonist binding. Receptor lipsda (agoniats or antapnists) will protect receptors from PCMB inactivation, but ths inhibitory process can r.lso be reversed by subsequent Irea/ment with organic sulfhydryla. Through the re- duclive alkylatioa of neurd membranes. Ndhy/makimide (NNM ) a n pre- vent much of the antagonist binding inhibition by subsequent PCMB trei.tment, but not that of the a4onist, suuestiaR that 11 la a mercuribenzoate, a ad not an elhylmaleimide residue within the bindiol sNe that interferes with receptnr binding. lfie NP.M treatment increases ajoeid binding by convertio` low agonist af8nity receptors to a.tate of high a6onial affinity, while their high dlinity for antagonists remains the same. The ability of NEM to Increase agonist tfinity is enhanced by (he presence of agonins, but not anutonisu, during the trealrnent. Low concentrationa of PCMB abolish the NPM ability to suA.equenlly increase agonist biodinL even when concentrations of receptor ligands sufficiently high to saturate the binding aites are included during Ihe P('MB treatment so as to protect the binding site. This suggests that NPM influences agonist binding through interaction with a group contiguous to the receptor binding site. 1)ependin6 on their concentrations heavy metal ions (Zn' •, C'd• •, C'o" and Mn•' ), which appear to interact with all of the NIiM- and PCMB-reaclive moeities, either increase or decrease agonist bind- ing, or decrease both agonist and antagonist binding. Different brain regiom contain membrane receptors that vary in distribution from the high- and low-atonist-a(finNy forms. The high-aflonist-afRnity muscarinic recepton are more numerous in the brain atem than in the kkncephalon, which contains more than the hippocampus. Roupton from different areas of the brain also have differing af/initiea (or sul(hydryl reagents and heavy metals. Aronstam, R. S.. Abood. L. G. and llow W. A(olerular Pharwrerolosy 14:S7S-SR6,1978. Other support: National Institutes of Health. From the Center for Brain Research, University of Rochester School of Medi- cine and Ikntatry, Rochester. N. Y. FNIIAN('1?MFNf (H cIPIAfI? HINI)INCI BY VAR1OtIS MOI P.(-111 AR 1ORMti (NU/•11OSPIIA I lt)YI 1FRINN ANI) INIIIBI I ION BY O1F11'R lINM1UkAtl 1) 111'1115 Nrr.e h..ue n t6..oth1 1.. .•.nr.m an opiate recelNot ealsUug as a prl+. lem •41d 1•h.•yh.dq..J. mrud•~.ur un.plca u( whwh phosphaedyherme. or i other acidic Iipid,'i. possibly an Integral component. Consequently, a wWe variety o( saturated and unsaturated phospholipida and fatty acids were ex- amined for their effect on the slereoepecifk opiate binding of rat brain Irynaptk rnembrane preparations. In fact, addition of acidic pho.photipids aiginific.ntfy enhanced opiate binding. Except for phosphatidyl.erine, "ic piwxpholipiaM containing a polyunsaturated acyl group were actually inhibitory aa were neu- tral pho.pholipids derived from brain. The CIe:0,22:6 form of phosphatidyl- aerine derived (tom synaptic membranes enhanced opiate binding by as mueh a 45%. Unsaturated fatty acids were highly inhibitory with tha degree of Inhibition related to the degree of uns.turation. Pho.photipue A and C wera inhibitory and the effect of A could not be prevented by albumin or overwme by the addition of phosphatidylserine. The use of dinilrodilluorobeorene, a eross-linkial agent, showed that the pho.phatidyLerine of synaptic roembranea could be preferentially associated with membrane protein. lAe enhancing effect of phosphatidylserine diminished with an increase is the degree of croas-linkiog. A6ood,L.G.rtalA elochinrka rt 8loohy.lra Acra 466:31-62, 1977. Other aupportr U. S. Public Health Service. Prom the Center for Brain Research and the Department of BiochemWry. University of Rochester Medical Center. Rochester, N. Y. I PHOSPNOI.IPID CNANOES IN SYNAPTIC MEMBRANES BY LIPOLYTIC ENZYMFS AND SI/BSEQUENT RESTORATION OF OPIATE BINDING WITN PIIOSPNATIDYLSERINE This study of the role of phosphalMyl.erinu in the xterempeciflc opiale binding to neural roembrana provides new evidel.oe in support of the thesis that phoaphalidylserine b an important, it not srrential, component of the opiate receptor. Among the signiflcant findings here .re the observations that: (1) exposure of mernbranes to phosphatidyhterine decarboxylase dimis- iahes opiate binding; (2) addition of phosphatidyberine following tredme.t with either phospholipase A, or Triton X-100 (inhibitors) wiB partially n- qore opiate binding: and (3) addition of phosphatidylserine following pptlal inhibition of opiate binding by treatment with phosphatidylserine decarboxyl..s will restore activity. lhe binding of 'It-naloxone, an opiate antagoniat, Is simi- lar to that of agonists in iU behavior towards phoaphAipaaea and pho.phatidyl- serine; however, binding of naltrexone, also an antagonisl, is far less respawr- sive. It is concluded thal the phosQhalidylserine associaled with the opiate receptor is the C1d:0,22:6 diacyl form, which is closely associated with protein. Abood, L. 0. at a1. eloch/mica er Stophy.ka Actts 3)0aS-46, 1975. Other support: 11 . S Public Flcatth Service. From the ('enter of Brain Research and Departmenl of Bwuhemistry. Unl- versity of Rochester Medicd ('enter, Rochester. N. Y. 52 411

i t HUMAN PLACENTAI. CHOI.INP.RO)C SYSTEM : STIMULATION- SECRETION COUPLINO FOR REI.EASE OP ACETYI.CHOl.INB FROM ISOLATED PLACENTAL VILLUS It has beta suggested that the permeability of the placental villus is coo- trolkd by acetylcholine (M:'h). According to a working hypotheuis. A/:'h iu released from the villu..timulating a eholinergk receptor on the nyncytiotro- phobtast, which kad. Io changes Ia the permeability of the trophob utic mem- braee; these changes are coupled to 1Se transport of nutrients acroas the trophoblut. Here, tbe conditions under whki ACb is rekaaed from the iso- lated placental villus were studied as a Anl step toward obtainiag evidence for this working model of stimulatios-ACA aecretion-tranaport coupling. Tha Molaled altus incubated Ie a muaele bath containing Kreb.-Ringer bkarbo- eale bufler apontaneously released )3 pmol/R/min of ACh Into /'x medium in a manner that was linear with 1ime. Tbere was no release ot .\('h In the absence of Ca" from the medium. With an increase of buAa G' • concen- tration from 2.4 to 4.61 mM, or with the addition of L-okotine (ftl rM) to the balh. Ihe rate of ACh release rose to 53 and 47 pmol/g/min, respectively. In the absence of Ca", however, nicoline did sot have any effe:t on ACb release. Atropine lowered both the rate of apootaneoui ACh rek ue and ib aicotine-induoed one, u did coeaine, a Car • antagonist; but d luhocurarioe (30 rM) did not aRect either rate. Tha presence of depolariziag concentra- tions of potassium In the medium (16-63 mM) also increased the nAe of ACh release. According to these dala, (1) the release of ACh requires tirc presence of ionic calcium in the ealernal medium; (2) Ca' • ions funcliou n a link between the stimulation of ACh and its Anal release; and (3) as ir.dicated by the effect o( nicotine on the placental release of ACh. there is a muscarink type of cholincrgic receptor in the ayncytiotrophoblast. Olubadiwo,1. O. and Ssrrry, d. Y. R. The Journal o/ rAornmroloty ond E, perLnentd Tl4eraperrlcs 204(2) :4l)-413, 1978. Other er.rport: U. S. Public Health Service. Prom the Dep.rtment of Pharmaooim, Vaaderbih University School of Medicine, Nashvilk, Teea. 711E INFI.UENCI'. OF CHOLINP.ROIC e1.OCKADB ON T11E UPfAK!? UF mAMINOI.SOHI('YRIC ACID BY ISULATI'.1) HUMAN PIaCI'NTAI. VILLI One of the major functions of placental trophoblast is nutrient transfer from the maternal blood to the fetal circulation. In view of the location of acetylcholine ( A('h )•chnline acelyltransferase (('hA ) in the placenta and the relea+e of A(-h into the maternal hkxxl, it has been suggested that A('h may stimulate a cholinertrc receplor un the Iropht.bla.1 and nNdula/e the placental tran.purt ut iom nd nutrienls In the e.periments dex'rihed here, n- 1"(-) 54 r I amiaoiaobrdyric aoid (AIS) wr used to aner the amino add uptak.e qnlnm ia human placenta and to measure the tramport efficiency of this active up- take system. Since it was not possible to obtain ACh-free human placer:ul villus, the effects o( agents which block ACb at various typea of cholinergic receptors and the effectc of antidroline.lerara on arnirw acid uptake by human placental villu. were studied ia this attempt to exptaia their antiRrowth effects. These studies inchded eholinergic bkockade by: (a) eaoeas e>togeaoua A(1, (b) massive release of endogeeao ACh by dcotiae, and (c) sparing eado- Renously released ACh by inhibitinR eholioeNer>t.e with pho.pholioe; also studied were the cholioergic blockade of: (d) trw.carieic reeeptor using atropine. (e) oiootink receptor of paRlionk type u.inR mecamylamioe, a.d (f) nicotinic receptor of newonw.cular-iunctbn type uaiag d-tubocyrarise. tinder all oondilioes eacept (a) and (t), Ih. AIB uptak. wr depressed by cholioergic blockade. '(beae ob.ervatio.n imply thN eadogenoualy reka.ed ACh eahibiu a mu.cariaic effect on placental villua and facilitates the tranr- port of amino acids. Blockade of the muacarinic receptor resulu in a deprea- lioo of the AIB uptake, a state which could, after a long period of lime, produce a retardation of fetal Rrowth. Rowep, P. P. aad Sartry, I. Y. R. Toakolop and Applied rA.rnrardop 13:79-9), 1976. O/her wrprtt U. S. Public Health Service. Frorn the Department o1 Ph.rmaeoloRy, Vanderbilt University School of Medi- ~ cioe, Na.hville, Tem. EFFECTS OF NICOTINE AND COCAINE ON THE RELEASE OF ACETYLCHOLINB PROM ISOLATED HUMAN PLACENTAL VIW The placental villu., which r devoid of neurnw.l innervation, w a.rw as an information wruw about the prooa. of aoetykholina (ACJt) rekau trom noA-nervou. Iitrues. With thi. In tnMtd, isolated human placental del were .u.perded i. Kreb'a bicarbonate bu//er ud the ACh relea.ed into 68 medium was analysed by gas chromatography. 71u .pontanear release of ACh into the medium was linear with time and was 35 pewl/R/min. Against this base line, K', Ca's. oocaine, and akotine were investigated for their inAuence on the pattera of ACh reka.e. Coeaiam In dwn of 290 and 560 rM decreased the spontaneous reka.e of ACh to 23 and 17 pmol/R/min, respoo tively. Raising the Ca' • concentration ia /he Kreb'. bicarbonate buffer (rom 2.34 to 4.64 mM or adding niootiee (SR rM) to the balh. Increased the rau of release of ACh to 53 and 47 pmol/R/min, respectively. Cocaine decreased the rate of release of ACh even in the presence of Ca' • or nicotine. Ilikh concentrations of K• increared A('h release in the presence of Ca• • bul It had no effect on A('h release in the absence of Ca ••; In fact,. A(lr was never released when Ca" was absent in the medium. Nicoline, which ie- creased both initial output and the rate of release of placental A('h, did not stimulate release of placental ACh In the absence of eaternal Ca•'. Cocaine 33

{ i depressed both of the observed nkotine-induced increases. These observa- lions indieale that (1) nkotine-ioduced increase in placental ACh is mediated through mu.carinic receptors. (2) nicotine iocreases The release of ACh by possibly facilitating the cell entry of external Ca•', and (3) cocaine de- pressa nkoline-induced incre..e in ACh rekue, possibly by depressing Ca •• entry into the oeli- Sanry, 0. V. lt., Olubadewo, l. O. and Boefirq, P. H. Arch/res /nternatfonales dt pAarmarotjn.A7lk tf dt TAlraple 229(1) :23-36, 1977. Oth...wpp.rt: U. S. Public Health larvb., From The Departmenb of Pharmaooiogy, and Obstetrics and Gynecology. VanderbiU U.ivenity Scbool of Medicir, NnhrilM, Tenn. HUMAN PI.ACENTAL CHOI.INB ACETYLTRANSFERASE. NATURE AND MOLECULAR ASPP-CTS OP THE INHIBITION BY IODO- AND BROMOACETYICHOLINFS /lalogenaled acetykholioes and other cholins oqen owe their high serum dtiolinnterase and nicotinic receptors aRinities so the eleclron dirplacement uused by the (nfluence of an Nubflitucnt. Organic reactioo studics have demonstrated that halogenalion produces sleric hindrance, and the s.me ef- fecl has been shown to have a significant role (n the hydrolysis of halogenated acy) groups by acelykholioesleraae and in their interaction at muacarinic re- eep/on. Their structural similarity to acetyp"C)choline (A('h), a product of the choline aoetyllransfer..e (ChA) reaction, mates the halogeru.acelyl- cholioes' ability to inhibit ChA and the nature of their inhibitions par/kularly important. The present inveqiption describes studies of halodenated acetyl- cbolina as inhibitors of human placental ChA. The resulting data support The formation of 1en.ry, inlermediates (CoA-ChA•IACh or CoA•ChA BrACh), during the reaction of placental ChA. The quaternary ammonium head and the positive carbo.yk.rbon do appear to contribute to Ihe inhibition of ChA by IACh and BrACh. An irreversible component of inhibitios (20-30%) ob- tained with hatogea-subatituled aoolykholines or lodo- and bromoacetnles can be explained by the possible: (1) formation of R<arbo><ymethylalyd t:'hA (where R- -0-, -S- or -N-), which is less active than The original ChA: (2) formation of Scarbotymethylated CoA, which is slowly released from the enzyme surface; or ()) eowvenion of part of the enzyme 10 the Inactive - N -earboaymethylated form. Ilcndenon. O. 1. and Sarrry, d. V. R. QloeAemkd tA.nnarolop 27:1331-1310, 197t<. OtAwr supportr U S Public Health Service. Nrt.m rhe lkparlmrnl of Nham.c.,luRy. Vanderbilt linivenity School of Medi crne. N..hville, lenn tt, r RELAT1ONSNIPS BETWEEN CHEMICAL STRUCTURE AND INHIBITION OF HUMAN PLACENTAL CHLORINE ACETYL- TRANSFERASE BY KETO ANALOOSOF ACETYLCHOLINE The /n rlro syolhesis of aeetykhdine ia achieved through a coupled aysl.m Involving an acetate-activatin6 entyme, aoelyl ooentryme A synlhetase, as,d an ensyme, cholioe aeetyltranaferase, that couples the activated acelata to choline. An inhibitor of the Anal step iu this synthetic pathway would, lherm- fore, be a valuable qent for studying cholinergic mech.nirna. le this aeries of esperiments, aeven teto analop of aoctykholine wero synthesized aed evaluated as lnA/blto.s o/ hrtrnae p/rantd cllo4nr aceryirrnwr/erara. Also tested was their capacity to inhibit horse serum dfolinesterasn aed to s14me late cholinergie receptors in the longitudinal muscle of the psinca pig. Of these aubNances, (2-bensoykthyl) Munnwoium chloride (compound VII). with an Ir, of 3 x 10-• M, was The most potent and sebclive bhibitor of choline acetyl(ransfer..e. Ito muacarink aad nieolinie sUmulatory activity wr quite low; so also was its cholinealerase-inhibilory activity. A study of Ih. Mruclure-activity, relatronahiys demonstrated three requirements for the i.hibi- tion of choline aeelyltramferaae by the altylaminoethyl eaten or their oocre- apooding quaternary aremordum derivatives: (1) a lermind eatioak head on the amine eod of the molecuk; (2) the ability to delocalize (or stabilize) a partial negative charge on The acyl end, and (3) a leaving group on The a-car- boa a1 the acyl end. Compound VII fulfills t.early all theae conditions. It has (1) a cationic terminal, a site for an electron acceptor interactlo.; (2) a. aryl moiety for hydrophobic and electron donor contributions; aod (3) a positive carbon atom adjacent to the bente.e rin6, due to the presence of The carboayl group, which interacts witL Me .udoophilic residue on the .nsyme. Cbalut.edi, A. K., Rowell, P. P. and Sarrry, B. V. R. lowrnd o/ pArnracertkaf Sclences 67(3):637-660, 1978. Other aupport: U. S. Publk Health Service. From the Departmenl of Pharmaooiogy, VanderbiN University Scbobl of Maal- dne, Nashville, Ten.. + (2-BENZOYLETIIYL)TRIMETHYLAMMONIUM CHIARIDE: A NEW, SELECTIVE AND STABI.E 1NIIIBITOR OP HUMAN PLACENTAL CHOLINE ACE7YLTRANSFERASE Since the discovery of .fyrYlpyridinea as the Ant poteM iahiblton of choline acelyltransfcrase (ChA), much effort has been devoted to the dovelop• ment of new and better ChA inhibiton of various types. Thr rrcport describa the propenks of a newly synthesized eompound, (2-benwytethyl)Irimethy4 ammonium chloride (BETA), a keto analog of acerykholine, which h s1abM aod meets The known structural requirements for ChA Inhibition. It was foued to be a selecrive, polenl, and noocompetitive InhibNor of (rA with respect to both of its sutntrates, acctylfoA and choline, with an 1„ of 3.1 x 10 a. Inhibition was rapid and slowly reversible, with 50% of the ('hA activity 57 -

i recovered within about 4.9 houn, upon dialysis of the inhibited enzymc. lhis new compound was also about 100 aod 50 times more potent In its ability to inhibit ChA thao acetykbolioesterase and cholinealerue, relpectively. It did not have significant activity at other cholinergic sites at the coscentrations at which It caused complete ChA inhibition. lu additioo to being considerably more selective than other ChA inhibiton, BETA is also more atabk. Pro- longed alorale of BETA solutions (several days at l7'C and p11 7.4) caused only partial and slow decomposition to trimethylamine and vinylphenylkel.me which was not suHicieolly rapid to eompronsiN Its potency as a ChA inhibitor. Rowell, P. P.. ('haturvedi, A. K. and Sanry,1. V. R. Ti1. Jornwl of PArmerolop enJ Ei>trMwMef TAeraprrrki 20S(t):624-6)4, 1978. OOther supports National Instilutee of Health asd U. S. Public Ileallh Service. From the Department of Pharmacolofy, Vanderbilt University School of Mcdi- eioc, Nashvilk, Tenn. P.FFP.CTS OP COO!_INO ON TIIP. LPVHIS OF ACElY1.CHOI.INP., CHOI.INFSI FRASP., CIIOl.INP. ACP.IYI TRANSFP.RASH AND Tll@ INTRAMURAL El.IiCIRICAI. STIMl11.Al1ON ON T11P.OUINP-A PIO ILEUM If the guinea pig ileum could be dener.ated by cold storage under anosia, it would constitute  suitable model for studying the effeets of atmisls that release endoge+aus acetykholine (ACh). Coosequently, the ACh content, as well as the cholinetterase (C1rE) and choline autyllranderase (CAA) activ- itin, of the guinea pig longitudinal ikal muscle were determined as a function of the duratioo of cold storage under anoaia. 7Le reaponsiveness of such cold- stored muscle to the Intramural electrical atimulation of Auerbach's pk.us, to direct ekctrical stimulation, and to etopenaa ACh were also measured. During the fint 72 hours of cold storage under ano:ic eonditiorn, the ACh content of the muscle decreased from 119 smol/I to a fairly steady kvel of 16 nmol/I of wet tissue. Concomitantly, the responsiveness to inlramurn./ ekc- tricat stimulation decreased by 72%, and the Ch8 and ChA activities each dropped by 40%. During this Ume, however, the de novo synthesis of ACh di4 not change dgnificantly and the response to esogenous ACh remained faitiy constant, suggesting that the decrease is responsiveness to Intramural stimulation is not attributable to changes in the ehotinergie receptor. Accord- ing to these dda. It is concluded 1hN after five day., cold storage (1) sig- nificantly diminishes ACh content within 72 houn; (2) decreases C'hPl and ChA activilks; ()) has no significant effect on the cholinergic receptor; and (4) lessens responsivenea to inlramural eketrkal atimulation, probably be- cause of a rnalfunclion of the AC'h-rekasing mechanism. lhis prephratinn seems to he a suiubte model for the differentiation of two types of chrdinu- mimetics. A( h releucra and direct muscarinic receptor stimulants 59 Ochillo, R., Rowell, P. P. and Sasny, d. V. R. PAarmacofoty 16:121-170, 1975. Other support: U. S. Public 1lealth Service. From the Department of Pharmacolo`y, Vanderbilt University School of Medi- eiae. Nashville, l'eso. ACTIVFTIPS OF S-METHYI.FURPURYLTRIMF'Tl1YlAMMONIUM IODIDE (S-ME111YLFURMEfH1DB) AT NICOTINIC RECBPTORS The (uras analog of muacarine, 3-methylfurfuryltrimetbylarnnwoius irr dide (S-me1hy1/urmelhide), reportedly atimulates eouscariaic receptors many thousand times nwre selectively than nicodak receplors. Other sludiea, how- ever, irdicate that it stimulates the parasympethNie ganglia of the guinea pig ikal longNudinal muscle; yet, this does not eceessarily prove that s-metbyl- funnethide haa nicotinic activNy, since like the sympathetic pn6lia, pus- srmpathetie ganglia may contain both nicotinic :nA muscarbie reeepton. 11ere, in order to determine whether 3-methylfurmethide stimulates the RanRliooio muscarinie receptors directly, the frog rectus musele was used to evalua/e the aetivities of this compound. The onset and reoovery of desessitkatioa, aa measured by depolari:ation of tbe poajusctional asembrane of the frog rectus abdominis muscle preparation following application of tbis aRent, show that the reaction is doae-dependeot sd that pbysoNipnine does not influence the doae-response curve. Repeated eaposure to a high dose of 3-metbylturmethide indicates that (1) the compound desetssiti>•es the cholinerSic recepors and (2) the onset of desessiti:atioa to the agent following eapourre to acetyl- choline is faster than the recovery. These results lead to the conclusion that 3-tnelhylfurmethide stimulates the aicotinic rooepton of frog rectua abdominis and demonstrate that high doses of this agent reversibly deseositize the recep- ton in this preparation. Since there is evidence iodicating that calcium Ia Is- volved in the onset and acceleration of desenaitiralion, aa appropriate eaplaea- Iion for the particular desensitization mechanism discussed here may require  flarough study of calcium ion movement during onset and recovery of the desensitized state. I Ochilb, R. F., Kau, S. T. and Sartry, i!. V. R. Pharmacoloskel RrsearcA CommwdeeUonr 9(e):719-727, 1977. Other aMpportr U. S. Public Health Service. From the Department of Pharmacology. Vanderbilt School of Medkine, Nuh- ville, Tenoessee; and the Department of Physioloq, Cornell Univenily School of Medicine, New York. • ARE MITRAI. CNI./ SCIIOI.INEROICI On the basis of recent observations of the anatomical dislribution, mole- cular propertics and drug binding activity of reecpton for .-bun4aroloais 59

(eBuTX), one may adopt as a working hypothesis that .BuTX is a nicotioic cholinergic rosrker in the central nervous system. An attempt was rr ade to test the validity of this aasumption by an analysis of rs'1-oBuTX binding within 1he rat olfactory bulb at the light nd electron microscopy levels, usirg both fresh cryostal sections and msteria) prepared by a combination of intraven- trkular losio injection and aldehyde perfuaion, followed by autoradiolraphy. The results of both methods were dmilar, and the auloradiographic patterns, which could be abolished try 10-' M d-tubocurarinc or nicotine, or a 50- to 100-fold esceas of native toxin but not by atropine (a muscarinic choainergic ligand), is.dicated that Ihe mitral oelV may be cholinergic, a proposition tested and supported by an experiment ora ncetykboY.aterase synthesis within mirral cells. These resulb are discussed here in light of other evidence concerning the chesnistry of synaptic transmission is the olfactory bulb. Accordinf to the aulhors, within the framework of our curreM kdowkdte concerning the neuro- tranmilter utilized by the mitral oep, it is possible to cJomlruct at kcit three snodels to accommodale these obeervations: (1) mitral cells are cholincrsic: (2) mitr.l cells are glutamste-ergic; asd (3) tnilral cells release both scclyl- choline and glutamate. Hunt. S, and ScAnsldr, I. Newrorcfance Syrnoosla 3:204-211, 1978. Otlser support: Natiooal Inslitulra of Health. From the Department of Biochemiary, State Univemity of New York, Stony Brook. SOM@ OBSf'RVAlIONS ON TItE BINDING PATTERNS OF a•BUNt:3AROTOXIN IN 1HH CEKyRAL NERVOUS SYSTEM OF T1113 RAT This report describes patterns of ar-buogaroto:in (aBuTX) binding with- io the rat brain and discusses their eorrelat'an with established and putative choliner& pathways. Tlx patterns delineated here were studied following one of these procedures: (1) intravenlricular injection of 's'Idabekd toxin fol- lowed by a survival period of /-a days before aldehyde perfusion, or (2) in- cubation of fresh cryostat sections of brain tissue in dilute solutions of radio- active toxin Appropriate controls with eicotine, eurare, alropine and native .BuTX established the specific nature of the binding. Principal ohservalions were that the toxin binding sites are predominantly asusciated with ccmral areas of the brain in direct receipt of sensory inpuu such as the main and accessory ollaclory bu11n, wperior colliculus, ventral lateral keniculste nucleus, and the dorsal column nuclei, and with limbic areas of the brain (hippo- campus, amygdals, olfactory tuberck). In many areas of the brain, silver grains were found to overlie cell bodies, suggesting that this may reflect the pre.ence of both memMsne M„rnd toxin and internalized ligand fn/iowing initul iamhnR lu a naiuhrane reaepN.r site. Ihe locahration of losin hindurt sdes currel.Irs rrasrmal.ly wrll wdh the aaon terrninals of previuusly dcscrdbcd r I chnlinerific pathways within the hippocampua, interpeduocular nucleus aad cerebellun. r Hunt, S. and Sclrnrldr, l. Aruln Retearcfi 137(2) :213-232, 1978. Other support: National Inslilutea of Health. From the Department of Psychiatry and Behavioral Science and Departmeat of Biochemistry, State University of New York, Stony Brook. T11B F.I.P('I7lON MICROSGOPIC AUTORADIOORAPHIC Id1CALIZA7lON OF a-BUNOAROTOXIN BINDING SITES WRl11N T1/E CNNI-RAL NERVOUS SYS7ZM OF THB RAT llse dislributiow of binding sNea within tM rat hlppocampua was Nudied after islraventricular injection of sssl-labeled toaim of hiilh specific radio.ctlv- ity. Animals were sacrifioed by vascular perfuaioa with rnised aldehydex aed autoradiography was used to show the localizatioo of the label. At the light microscopy kvel, the label was found to be concentrated in a earrow b..d within the outer portion of the slralwn oriesr. a. area of termination of the cholinergic mediai-septal hippocampal pro;etyion. At the ultrastructural kvel, fabekd Wain was found .InaN esclusive/y, is etrodalios with synaptic oors- pkttea characterized by atrymmetric contacts (Or.y Type 1) and ckar, spher- kal, moderately packed synaptic veaicks, cas .eea in known cholinerBic juuo. tiom. 1s cryoslat materitl, binding of toain could be suppressed by low cos- centrations of nicotine and d-tubocuraritse, but wot by atropine. Taken 1o- 6ether, these observations suggest very slronRly that .-bunprototin binds to uicolink acelykholine receptors located on interneurons of the Mratum oriaoa, and that poslsynaptic truke to><itu cae be used s specific probea, not only for acetylcholine receptors is peripheral rseuroeffector iunclioaa but also foe aicolinic receptors in tha central md autotsomic eerrar system. HtuN, S. P. and ScAndit, l. ardn Rerearch 112:152-159, 197s. Other er. pport r National Inqitutea of Health. From the Department of Psychiatry and Behavioral Science, and the Depari- menl of Biochemislry, State University of New York, Stony Brook. DISIRIBIII ION OF w-Bl1N('•ARO/OXIN BINDIN(7 SI7'FS IN TH@ CI'sN I RA1. NERVOUS SYSfNM XND PNRIPIfNRA/. URUANS OF '111E RAT Biochemical lechniques were used to determine the concentrations of .-bunssrolosin binding sites in various regwns of the central nervous system and in several peripheral lissues of the ral. lhcre were marked regional ddfer- ences within the hrain. No activity was delectcd in the caudate nucleus and little in the cerebclluns; the interior and superior eolliculi and the hippocampw 60 61

demonstrated the highest levels while intermediale densities were ob;ained io other areas. Oulside of the central nervous system, the binding activity was sibniflcant only in skeletal muscle, autonomic ganglia and adrenal btands; some ar-bunbuotoaia rnccpon were :ho found in sensory 6anb/ia (Gassetian and dor+al root). These results subbest that ~bunproloaio is specific for nicotine ace/ykholine receptors regardless of Iheir location within the organ sm, and th.t these receplon are not necessarily ayeapek oonnitueob. Jcbechter, N., Handy. 1. C., Peu.assd, L., aad ScAmldr, !. Totkow 16:215-231, 1971. Ot/1er esspport: National Sckoce Pousdatioa. From the Long Island Research Institute and Dep.runeet of Psychi.rtry and Bebavori.l Scknee, and the Department of Bbchemistry, Stato Uaiv.rsity of New York, Stony Brook. I.IOAND RPSPONSFS OF Q-8l1NOAROT'OXIN BINDING SIIFS fROM SKk1.ETAL MUSCLE AND OPTIC LOBC OF THB CHICK Binding properties of deterbentiolubilirtd receptors for rbunbaroloaio from skeletal muscle of the 1l/h day chick embryo and from optic lobe of the hatching chick were compared. It was found that both types of receptor are nicotinic although they differ in their aflfnilin for Individual ligands and in the rank order of ligands. In contrast to the muscle receptor, the neuronal receptor hinds the tosin in a reversible fashion (Ko - 2.1 x 10'sM at 23'C). Small IibanJs inhibit receptor-toain auociation, as analyzed by kinelk and equilibrium procedures. To.in and ligands compete for a single type of noninleracling site, and the ratio of losio binding sites to lieand-binJing sites Is unuy. The inhibitory potency of ligands parallels their ability, at higher concentrations, to accelerate receptor-/otin complex dissociation. llria accelerating effect is not saturable. It is proposed that nicotinic IigsnJe block association and induce dissociation of toxin by interaction with the same site on the receptor derived from the optic lobe. Wang. O-K.. Moliaaro, S. and ScAm/dr, 1. TAe lournd o/ Qiolopkd Chembrry 237 ( 27 ):l307-lS 12, 1978. OOther .rpport t Natiooal Science Fouodatioa. From the tkpartment of Biochemittry. School of Basic Health Sciences. Slate University of New York, Stony Brook. V. Ph.rmacolosy and Biochemistry 7 RAPPINO ANT) 1)P.TrRMINAlION OF LABIl.t3 COMPOUNDS IN IIIF GAS PI/A%I: . (11• (-1(:ARI 1-1 N SMOKK In ihs. •tudy. ttnaa sdrnplwn fullaweJ hy Ihtrmal de u.1pla.o s1../ ta. .h~.nn.~nRr~l.hy war cvaiu.ltd a• a way of tharrttrruins r the volatile orpniti pa phase constituents of smoke. Speciflcdly, the highly labile compounda iaoprene, aoetaldehyde, and acrolein were dekrmiued quantitatively in the samples which were thermally desorbed in the injector pott of a 6as chromalopaph onto a cooled Baa chromatographic column. All of the isoprene and acrokin and 80% of the acetylaldehyde were recovered. The pattern of the chroa•.alobranu indicated that other m: components la the gas ph.aa wars r.oove:Nd also. Study of the allecta of a~ on the trapped gases showed that improrements eced to be made In the storage of aged aunptes. Given the.e, thir method should be appllcable to remote auoplia6 of cigarette smoke and other psem for subsequent andyae" at .oolher aite. Zeldea, S. O. and Horton, A. D. (Unbn C.rb/de) wwdytkd CAendtery 30(6):779-7/2, 1978. Oth.r etrpporti Natiosaf Caacer Institute and the DepartmeW of Merfty. Frorn ehe Analytical (bemiatry Divbioa, Oak Ridge National l abocatory, Oak Ridge, Teon. I ROLE OF TOBACCO SMOK6IN PHARMACOKINETICS Tob.eca, which it widely used ia our aoekty, can alter the dispoaitioa aad pharmacological eflecta of awaP ootnmoa drugs and ahould, therefon, be looked oa as one of the primary aouroa of drug Interactions In man. In thia review, the role of tobacco smoke in pharmaookiaetica and clinical pharma- cology is emphasirtd, with aeclioas of Ihe paper examining (1) the disposition of phenacetin, antipyrine, asothinea, and other drugs; (2) drug effects ia man; (3) mechanisms of lobaoco-drug intersctioa.; sod (4) nnoking sad drug consumption. Moat of the experimental worR ia aun, animds, and aww or enzyme systems indicates that aha dominaM eQect of smoking is enhaeeed disposition caused by inductioa of hepatie wieroaornal eniymes. The primary causal agents are probably the polynuckar aromatic hydrocarbom, which an potent and persistent in tissues. Besides the ideatilkation of a wider array of drup, enzymatic pathway* and dioicd eAects that are altered by smokia& further HuJies are needed to examine the role of snwkiag and its poadbla eAects on other ckaranea processes ia man. While several of /he hepatic mkrosomal drug mctabolixlnB erwymca are stimulated In seakera, the seloc- tivily of this reaction is unpredictable and the effects of tobacco tanoko on other potential rate-limitins disposili.n processea are largely unexplored. lrsRo, W. !. lorrnd oJ Phnrmarol/netkr and dlophmmorertks 6(1) :7-)9, 1978. Ot6er supportr National Institutes of Health. From the Department of Pharmaceulics. School of Pharmacy. Slate llnivenity of New York, Buflalo. 62 61

PhENYTOIN DISPOSfT1ON IN SMOKERS AND NONSMOKERS In this attempt to esamine the role of smoking in Iba disposition kinetics of phenytoin in man, each of 16 young adult volunleen (It smokers and 8 nonsmokers) received a 3.6 ms/kg Intravenous doae of the low therapeutic indes drug over a 5-minute period. Blood samples were collected before and at 0.16. 0.5. I, 2, 3, 4, 3, 6, 7, •, 24, 32. 48. 56 and 90 houn after dosint, aod plasma concentrations of phesy/oia were determined by ps-liauid c:xoma- tography and radioimmunoassay tecb.ique.. Results showed that smoking did not affect the disposition kinetics of tM iMravenouniy administered phenyloin. Nonlinear least squares computer U of the plasma eoncenlratios-time data yielded coeflicients of a biesponenlW equation for cakulalioe of hslf-lives (tw), ekaranees (C1a) and volumes of distribution. Thers were no sla- tistkally signifkanl diRerencea is tAerr paramelen with respect to either set or smoking However, eate variable tyjs were noted in the smoking group. No diRerencea In serum protein binding bNweea smoking and nonsrnokln6 groups were found. Serum Ihiocyanate concentrations were used as a neasure of smoking habil; there, was no aipsifkant correlation belween thes/e thio- cyanate concentralbns and /ty or Cla. Attempts to correlate phcoytoin Ct, with basal metabolic rate also failed. Rose, 1. Q. Barron. S. A. and lrirAo, /Y. !. )nternerlond lorn.d of CYlnkd tllarnrorolop .n1 dioPllarnsory, 16(11) : 547- 330, 1978. Other erpportr The Jacob. Family Foundation, Inc. Prom the Clinical Pharmacokinelica l.boraNory, School of Pharmacy. State Univenity df New York at Buffab, and the lknt Neurological Inslilute, Millard Fillmore No.pital, Buffalo. SYN771ESIS OF 4-METHYLNICOTINE AND AN EXAMINATION OF fIS POSSIBLE BIOSYN7NESIS FROM 4-METHLYNICOTINIC ACID IN NICOTIANA TAeACUM This lobaooe ehemiatry article deacr(be1 attempta made 1o produce 4- melhylnicotine by .dminiqering 4-metbylniootinio acid to the tobacco plant. In the flrst of 1hree preparative rnelhod., oondeorNioe of ethyl 4-melhylnkotinale with N-melhyl-2-pyrrolidone in the preseeoe of sodium hydride yielded the .iealinoyl derivative 6•, which on hydrdysis asd reductioa afforded 4- methylniootine (8). TAi..iootins andosue waa abo ob/aioed from 4-melhyl- pyridine-3earboaaldehyda by preparing the acy) carbanion equivalent 2, which was added to acrylonitrik. Finally, hydrolysis of the Michael addition product yielded Ihe ketonitrik 3. which was hydrogenated /o give 4-rnethylnornicotine, which afforded I on methylation. Other experiments showed that a biomimetic synthesis of ti involved reaction between 3-melLylglutaraidehyde. ammonia, and N-methyl-e'-pyrrolinium acelate in the presence of air. Optically active f; was ohlsineA, as previotnly de+eribed, by reaction of (-- 1-(2"S) -nicotine with mclhyltilhinni, and the adminnlration of 4-melhy114-"(')nicotinic acid (pre- paie.l Ili-m rlhy111 '4 I.1aetaatelalr) lo Niruriano raAorurn plants diJ not rr%ull In 1hr b.rnulu.n ul rs-hnatlore 4 mclhyln+ct+thne. When tested pharma- A4 t oologically, 4-methylnicotine showed no antagonism to the stimulating elfect of oorepinephrine; the compound showed .o alcotlne-like activity in tbese le.ts. •Numben used refer to the compound'a position ie an illustrated achearo of eicotine analogues. L.eere. E. and lsele, S. A. S. lorrnd o/ Organic Chemistry 43(11) :2122-2123, 197t1. Other ar pport i National /ostNulea of HeahR Prom Ihe Natural Products Laboratory. School of Chemistry. University of Minnesola, Minneapoli.. SYN771ESIS OF A `BRIDOED NICOTINE": 1.2,3,3,6,10b-II N.XAIfYDROPYRIDO(2,3,t/INDOLI7INH Although many analogues of niootine haaa been prepared already aad their pharmacology studied in an attempt to ob.fs Mruclure-activily relatioa- ahip., no work has been roporled yet on the preparation and pharmacology of bridged nicolinea. Interest has been esprered in these compounds, how- ever, and this paper describea the syahesa of the bridged nicotine 10 aad alro 2-metbyloicotine (3) by the roula illustnted is Scheme 1: St:beme I a cjs ~ I. qwr. _ ., / r 3 4 0 0 a 0 to 7Le pharmacology of 5 and the bridged nicotine 10, a pyridoindoti:idine. Is being examined now. 6S

Catka, T. F. aod itere. E. lorrnal o/ Orsanic Chcmfrtry 1)(11) :2125-2127, 1978. Other support: Natiooal Institules of Health. From the Natural Products l.ahoratory, Scbool of ('hemislry. University of Mionesota, Minneapolis. PARI7CIPA7ION OF PYRIDINO-N-OXIDFS IN THE METABOLISM OF NICOl7Nl? IN VIVO - A PRF1-IMINARY STUDY There is a possibility that pyridino-Noaidea corresponding so a dozen or more metaboliles of nico/ine may participate in the metabolism of nicotine. To date, Mrwever, little deflnitive informatioe eaiats in this area because of analytical difFcuhies in eslablishinit the presence and quantity of Nosides sl low levels and a lack of unifying principles upon which to predict pharma- cological activity of these substances. In an espbration of the Noside problem, chromatographic data were collected on urinary basic metabolites from rhesus monkeys alter oral administration of (S)colioine (570 m6/kil) and (S)- cotinine-N•o.ide (SlS mg/kg). The results suggested the presence of a number of Knenig-posilive "basic" components, including colinine and the cotinine metabolites )-hydrosycotinine, demethykolinine, and allohydrosycolirnne. Con- trol urine of monkeys prior to the initiation of feeding eaperiments with S-cotinine and with Stolinine-Noaide showed none of theae Koenig-posilive components. The close correspondence In distribulion of Koenig-positive sub- stances in the urine of the treated anirrrab suggests that there is a biological equilibrium between colinine and tblinine-Noaide in the rhesum monkey. In a similar series of eaperimenls, chromatographic examination of rabbit urine after administration of )-pyridyl.cxtie acid-Noaide provided evidence for the escretion of 7-pyridylacetie acid and N-(l-pyridyla«tyl)glycLne. Both 7-pyridylacetk acid, a metabolite of nicoline, and N-()-pyridylaoety/)glycarc, a metabolite of 7-pyridylacetic acid and hence nicotine have been pre- viously studied tor their ability to block some nicotine-like effects of rrans- melanicotine on aorlic strips. Since )-pyridylacclic acid arises from she metabo- lism of tranr-metanicotine, N-(7-pyridylacetyl)glycine and possibly other con- jugates of l-pyridylacelie acid are implicated /n the metabolism of inetanicotine. McKennlr. lf. lr. et of. In: Oorrod, 1. W. (ed.): abfoticd Osfdotlon of Nitrogen. New York: 1'.lsevier/ North-Nolland Biomedical Press, 1978. pp. 163-16E. Other support: American Tobacco ('nmpany. I r...u Iht 11..r.1un .d H~.rhr~..,.al /'hs:rnu.u6ogy. Mrdc.l ('odlege (if VnKinu, W1.1 ,,,.,1.,t I Mf:TABOI.IC N OXIDATION OF 3-SUBSTITUTED PYRIDINES: IDENTIFICATION OF PRODUCTS BY MASS SPF.CTROMETRY Many of the basic constituents of tobacco and tobacco smoke contain a heteroaromatic amino group; but in spite of the eatensive occurrence of this type of mokcuk. the biological o.idation of Its constituent nitrogen has not been widely observed. Technological diflkullies, mainly created by the properlies of these compounds coupled with the limitations of conventional rnethods, have probably prevented the detection of these compounds in pre- vious meUbolic studies. However, the presence of an (M-16/! ion peak ob- tained upon mass spectrum analysis of several aromatic amine N-oaides was considered of diagnostic value. It now seems evident that this ion is formed solely by thermolysis. This report describes oonditiom that minimize thi. lhcrrrwslysis and allow masa spectrum and gaa liquid chromalography-masa spectrum analysis of various )-wbstiluled pyridine-N-o.idea. The mass spectral characteristics of their N-oaides arn deflned. MoM of the lime. the base peak was (ound to be the molecular ion, provided that thermolyais was minimised when using the direct probe or gas chromaloRraph inkN. The chromalopaphic and mass spectrum evidence presented here also indicates that, both in rlro and in ritro, the biologic osidation of the beteroaronutic nitrogen (n )-subdituted pyridines is a metabolic event whose pathavay has quantitative importance. Cowan, D. A.. Damani, L. A. and Corrod, l. W. Olomedicd MarsSpectrornetry S(9):SS1-SS6, 1978. From the Department of Pharmacy. Chelsea Colle6e, University of L.osdoa, L.ondoo, Eogland. DIRP.CT OAS-LIQUID CNROMATOORAPi11C ANALYSIS OF SOME PYRIDINti-N-0XIDES Tertiary aliphalic and aromatic N-osldes are Ihermolabik and their structure is altered during gas-liquid chromatography. Consequently, their analysis has usually involved complicated methods. Because aromatic No.idea re inherently more slable, however. this study attempts the direct analysis of some pyridine-N-oaides by gas-liquid chromatography. A technique for the selective eatraction of these compounds from biological materials is also described. The procedures used here have made possible the development of sensitive techniques for the direct detection and quantilation of alkyl- and haloken-subs(ilulcd pyridincs-NoaMks formed during fe vitro or !n rfro mcla- M>tk sludies. Neverlhekss, espcrkecc Indicates that careful manipulation of instrumental conditions may be neceasary in order to ensure minimal decompo- sition of the compounds. Corrod, J. W. and Damani. l.. A. Journal of Chroniarog.aph y 133: 749-75 ), 1978, From the Department of Pharmacy, ('helsea College. llniversily of I ondon, I ondon, !?n`land. 67

THIN-I.AYER CHROMATOORAPHIC SEPARATION AND 11)1'.NFIFICATION OP 1 ERTIARY AROMATIC AMINES AND l l(I:IR N-0XIDES The metabolism of a range of simple structural types of nitrogenous compounds was studied in this attempt to throw IigM upon the occurrence and en:yrnoloRy of metabolic N-oxidation. Piperimentally, thin-layer chro- malographic (TLC) systems for the separation of •-wbstituted N<thyl-N- methylanilines and their N-dealkylatioa and N-oxidation products were em- ployed. Various T1,C systems fur dre separNion of 2-. 3- and 4-subslituted pyridines and their Nosida were also sraed. Most of the detection reagents chosen for this study were standard cobr reagents that are used mainly for detecting aromatic amines and phenols. Resulta showed that none of the re- agents used here was specific for the detection of the pyridine N o.ides on ll.l.' plates. Howcvcr, e.amination under l1V light at 254 nm coupled with their characterintie R, values In 711,47 solvent systems proved a succeu/ul melhod for the idenlifkalion of microtram amounts of these N-oxides in biological fluids. Tetracyanoelhykne (TCNB) was found to be a very aensi- tive and specific spray realent for the detection of N,Ndialkylaniline-Naxidd, and the results obtained with TCNP and reduced aodium nitropruuide indicate that they may be of use In the detectioo of aliphatic and alicyclic N-oxides. Damanl, 1.. A., Patterwn, 1.. I(. and Corrod. I. W. lorrnd oJ CAron+crotrophy 1 SS:I)7-)4S, 1978. Other support: Science Research Council and Chelsea College (University of l.ondon). Prom the Department of Pharmaey, Chelsea C.ollege, University of Inndon, tmdon, England. ON THB MULTIPLICrT Y OF MICROSOMAL N-OXIDASP. SYSTEMS llre work leading to the recognition that biological oxidation of nitrogen in organic molecules probably involves more than a single enzyme system is reviewed The attempts to differentiate these processes and to correlate them with the pKa of the susceptible nitrogen are described, together with recent work on the enzymology of two distinct tertiary amine osidase systems. llre results are taken as further evidence of diRerences in the enrymodosy of N-osidation. 'llre actual mechanism@ invdved still remain to be del:rmined, however. Whether the nitrogen is oxidized as a result of 1he formation of supero.ide anion during the breakdown of flavin peroxide or wheth:r direct o.idation is involved through Ihe Aavin peroxide itself ia not presently known. Another possibility is that aliphatic amines form a tertiary eomp:ex with flavoprotein and molecular oayten by the donation of a lone pair electron. Additional work is needed bcfore the nrechanism of nitrogen oaidNion in orjanic nrulecules cui be elucidated. (:nrroJ. 1 N'. I In: Sin=er, T. P. and Ondarza, R. N. (eds.): iNeclianirnu o/ Oxididnt Ew- zymes, New York: Elsevier/North Holland, lnc., 1975, pp. 1t9-197. Ot6sr supports Science Research Council and Ihe Cancer Research C.rn- paigo. From the Department of Pharmacy. Chelsea CoUete, University of Londoa, London, England. Ti1P 1?FFP.CTS OF NICOTINE ADMINISTRATION ON DP.CIDl/OMA INDUCTION IN 1118 RAT Nicotine modifies the implanlation process In pregnant rat. by delay!ol the development ut Alastucysl attachment. 71ria may be a result of changes in uterine receptivity to the conceplu. (which is responsible (or the inductio" of deeiduatizuion) brought about by modifications ie prolaetin and propa- Ierone secretion. This study attempts to determine to what degree. (f at aN, nicotine modifies decidualizalion in the ral. Paeudopregnant animals derrwn- s/raled impaired decidualization following subcutaneous nicotine injections. The greatest inhibitory effect seemed to occur during the first four daya of pseudo- pregnancy when uterine sensitivity to decidualization is developing. The data also suggest that nicotine may impair decidual cell growth either directly by acting at the cellular kve(, or indirectly by .wdifying bormonal kvels, i.- dicding the need for mwe detailed studies in this area. Neverthekr, ths degree of uterine raponsivNy suppression Induced by nicotine is dosedepend- ent. While this report clearly indicates that aieotine modifies uterine funqio, in the rat and is thus useful for studying certain aspects of reprodudive physioloty. the authors emphasize that the results cannot be extrapolated to women who smoke. Card. J. P. and M1rcAelf, 1. A. eiology oJ ArproAucNon 19(2): 326-331. 1978. From the Department of Anatomy, Wayne State University School of Medidea, Detroit. AI TERATIONS IN SURFACE MORPHOLOGY OP THP. ANTIMF_SOMIiTR1A1. UTERINE E'PITHFI.IUM OF 71fP, RAT: L'FF@CIS Of' OVARIAN STEROID HORMONNS Not only does nicotine possibly modify blastocyat-uterine interactioa, bul it also decreaaa uterine respunsiwty to deeidualiration. It ia conceivabk that these nicoline-induced changes may result, al k..t in part, from nicotine mod- Hkation of estrogen and/or projea/erone secretion. Accordin=ly, changes in uterine luminal surface morphology were observed by scanning electron microscopy during the atrous cycb, and the effects of ovarian steroid hormone ~Ieprivation and slimulation on the endometrial surface structure were nolod. OvariecNxny produced an epithehal surface morpholugy similar to that aoen during diestrus, but more prorwwwnced. t•stroden replacement produced changes similar to those characteristic of estrus that became apparent within 24 houn f, N /GO

of lreatmcnt. In another group of ovarieclomiud rats, progesterone caused dense, short microvilli and numerous roueh-surfaced protrusions. Tbii study confirms previous reports concerning the marked ovarian steroid hormome de- pendence of the endomelrial microvilli, uterine glands and uterine secrelory, aactivity. It also demonstrates that scanning electron microscopy is admirably suited to the rapid eaamination of ealensive luminai surface area.i, thus facilitating the study of surface nsorpholop and the assessment of both normal and eaperimentally-induud changes is surface structure. Hammer, R. B., Samarian, R. aad AlfleA./,1. A. Scanning Efrcrrow Alirroscooy 11:701-706, 1978. Prom the Ikpartroent of Anatomy. Wayne State University School of Medicine, Iktroil. Vi. Immunology and .Idaptive Mee/ranLnsa IMMUNOC7I.OBU1 IN A IN SP,CRP.iTONS FROM Tlll? LOWER HUMAN RI.SPIRAIORY TRACI' Studies on broncho-alveolar lavage (BAI.) material obtained from 45 human voluntcers, both smokers and rsonsmokcrs, show that I&A represents about 5% of the pratein content of BAL. The free secretory component (s-IgA) represents ahout 1%, this proportion remaining the same retardku of the smoking status. IgEi is also present. There were major differences, how- erer, in the composition of immunotlobulins at various levels of the respira- lory tract. Tlsis has previously been systematically documented in a canine model wherein the ISA to ISE ratio was alao found to ehange, and il may indicale relativs functional differences in the importance of these immuno- gbbulina al various levels of the airways. Based on their own d.la, the sulhots conclude that whik s-ISA is not the mosl opsonic antibody in the lower respira- tory Iracl, this does not preclude other important functions for the molecule. Although its most important role is in the oropharyns and upper airways, s-ISA may also be critical in preventing the ciliary attachment of mycoplasms organisms aad viral agents along Ihe epithelial surfaces of the /rachea and bronchi. However, its antibody activity may be ku useful in the alveoli where phag.,cylie cells appear to be the first line of defense against inhaled particks and infectious agents Ihus, in spite of Its relative ineRecliveness as an opsonin, s-IgA may neverlhekss have a major function in the ttspiratory tract through its regulatory role on antigen entry and the dispoaal of inhaled envirunmenl.l and microbial antigens. RrynalJr, if. Y. et af. AJvun,rr ln 1•.prrimrnrd AIrJorine anJ fliofogy 107:SS1-Sh4, 1978 Fmm ihw -1'adns.ina.y tiecuon, /)eparunent of Intern.l Medicine. Y.Ie l)ni vew.ow tik hiw ol nl M. Jw ine. New llaven, ( unn I IMMUNO(7LOBOLIN SECREI7NU CELLS IN NORMAL HUMAN BRONCHIAL LAVApE FLUIDS As an Immunocompetent organ capable of an antibody response so pv- liculate antigen. the lung contains cells with ir.rnunolotic capabilities through- out its parenchyma, bronchial lymphoid drue .od diual bronchial-alveolar airways. The present study uses a modified reverse hemolytic plaque assay lo aaseas the immunoglobulia secrelory capacity of normal human bronchial alveolar cells. Tbo...ecreling inum"lobulim were quantitated in the brosa chial lavye fluids of 12 normal volunteers and compared with the otsa found 1n peripheral blood. The lavage euids contained a relatively higher tnuaber of both 1S(l- and ItA-secreting cells than peripheral blood. llowever, the I&A- aecreting cells were the preponderant type in the Ruids, while ISO-socrdfag cells predominated In the blood. The mean ratio of IgA-/1CO-secretlng caYs was also higher la the lavalie Ruide than In the per.ipheral blood. Only four out of seven lavage samlAes studied coetdned 16M-secrelinR ulla, but tAae were found in all peripheral blood samples. By conlrasl, normal peripheral blood did not contain any 16Esocrelin6 u8s, while Ihese were demonstrated In eve out of I 1 lavage specimens. The normal human lower bronchial-dveolar Iree, therefore, contains cells that actively aecrele irnmunofilobulln.. I/ is sulp- gested that those .eereting 1S0, IsA, or 1sM may iuncUon in local pulmonary immunologic mxhanisms, while the onea seaNinR 16H may have a role 1s hypersensitivity reactions in the king. Nererlhe/na. the ability to distinguish class-speciAc immwa6lobulin-t+ecretins icell. in bronchial lavage fluids adds a new dimension to the functional andysis of itmmmoeompelent pulmonary cells. Tlsis type of technique may mate It possible to Identify the local pulmonary immuoore6ulalory mechanisn» at work in normal and diseased lunp. Lawrence, B. C., Blane, R. M., MarNn, R. R., a.d Stevar, P. M. TAe lownd o/ t7inkd Iwre><Heaion 62:t12-tlS, 1976. Other.apportr Natioaallnstiluta of Healt6. Prom the Metabolism eru.ch, National Cancer InuituM, National IrMd.e of Heallh, Belheda, Md.: and the Departmenu ot Medicine aod Micsobiolop and Immunology. Baylor Colle6e of Medieine„ and The Methodist Ho.pkal, Teau Medical Center, Houalo.. NORMAI. HUMAN BRONCHIAL IMMUNOOLOBULINS AND PROTEINS: EFFECTS OP CIOARETILr SMOKINO Selected proteins in the bronchial secretions recovered from 36 healthy nonsmokers and 19 healthy cigarette smokers were compared here N an at- tempt to determine whether eiprs/te smoking aller. /he composition of 1bw pro/eins. Although /luide Isva6ed from smokers contained four times u many cells (more Ihan 90% m.crophaees) as fluids from nonsmoken, there were no quantitative differences between the Iwo groups in immunoelobulin A, &,-.ntilrypsin, e,-globulin, Iranafertin, or albumin content. Iloweve , the total amrwnl of immunoglobulin O obtained from the lavage fluids of smokers was twice that recovered from rKrnsmokers. In addilion, there was a/endency toward higher concentrations of albumin and total proleins in fluids from smoken, but these differences were not signifkant. More than half of the 70 71

oeeretiaos from smoken contained traee amounts of occult blood, ~rbe:reas only about one fourth of the aecrelioos from ooosmoken were positive. Tbele w•s tw correlation within each group, however, between total protein or im- murwglobulin 0 content and the presence of a positive test for occul: blood. Allhouith the mechanism behind the Increased concentration of 1t0 noted here has not been estabinhed, this selective inerease In immura6lobulia could be compatible either with stimulated local bronchial immunoglobulin produc- Iion, or the selective exudation of planm• Ismuooglobulin into alveolar spaces in response to inhakd cigarette smoke. Wan, U. A., Mertln. R. R., Sharp. P. M., and Romen, R. D. Amrrkan Review oJ Rrrplrarory Dlsesr 116:25-)0, 1977. Other arpprtr National Institu/a of Hedtb and the National Aeronautia and Space Administration. From the Dep•rtment of Medicine and the Department of Microbiology and Immunology, Baylor College of Medicine, Houston. SYNTHESIS AND RELEASE OF THYMIDINE BY MACROPHAOES Previous experiments have shown that culture fluids from cell cultures rich in macrophages contained an Inhibitor of DNA synthesis which, when aasayed on cultured thymic kukemia ulls (line EL-1), inhibited thcir growth ond prevented the incorporation of radioactive thymidine into their DNA. This growth inhibiliun•was casily reversed upon remov1 of the inhibitor, which was characterized as a small, dialytabk molecule resistant to phosphodicsterase treatment aod capable of adsorbing lo the lumw cells. In this study, the in- hibitor ftom macrophqe wpernatasu was determined to be thymidine as established by thin layer chromatography and 6e1 filtration. lAis inhibitor was also shows to be synthedred de novo by maerophajes pulsed with precursors of thymidine eoch as "Cdormate. The amount of thymidine present in lissue culture fluids may compete with radioactive tbymidine and ia suB'icient to block the growth of EL-4 ulb in a manner that could be prevented with the addition of 2'-deoaycytidine. Theae results indicate that tnacrophapes secrete thymidine continuously a• p•rt of an active ayethNic process. Most likely, other nuckosides are produced as well. The •iRniticance of such synthesis and excretion of thymidine in/o Ihe outer milieu ia uotnowo, but appears to be unique for macrophallea. Consequently, aulioa should be used when inter- pretind data from culture aystena in which macrophapes or their products inhibit cell proliferation and/or DNA synthesis. The capacity of entire super- satants to affect cell proliferation aeems to be dependent on two critical para- melen: (1) a balance between inhibilors sod slimulatory mokcuks. and (2) the netwre of the target cells. The EL-I line b especially prone to inhibition by tbymidine compared to a number of other cells tested. Sladecker. M. 1., Calderon, 1., Karnovsly, M. L., and Ununue, E. R. Thr /ourna/ o/ Immunology 119131:171s-1747, 1977. Othrr w/rers: Natweal Insthturrs of llcdth 1'rum the 1lrparuui ettr of P.thology and Biological Chemutry, Ilarvard McJn.1 1.hout, N...t .n I i I REGULATION OF LYMPHOCYTE PROUPERAT7ON AND DIFFERENTIATION BY MACROPHAOB4 One maior function of the maerophqe b modul•tion of the inler•ctbn of T cell, 8 cell and antioles, whJe a second characteristic lunetioe of this cell in its aecretory activity. Rooeetly, a. eap•ediog lilerature has appeared that unites these functions eonoeptua0y, invealiptins the secretion by mncro- phabea of lymphostimulatory products that modulate various p.runeters of irnrrnrne function. In 1Na light, the present paper examines (1) the rote of a macropha6e diRereotiation, •nd (2) a matropba6e-iramrwe T cell inleracaioa that results in augmented necretioe of lympboalimulalory, tfactors. When eu4 lured with ebe thymocytediRereotiatio6 factor (TDP), tbymocytea show a physiological increase in the major histocomp.tibilily taolecuka. H-2a andd 11-2s, decreased aensitivily to lysia wUb aoti-TL •nd eornpkment, and •equisi- lion of responsivcneaa 1. 1he mixed lymphocyte culture. Development of the mature phenotype requires two to tlwee days of eultwe aad, onco attaioed, is stable. The ialuced anti6enic changes do aol require cell division. The activity demonstrated by 7DF, whicb is not •rtributabk to ioterferon and cannot be replaced by 2-mercaptoetbanol, is also displayed by normal thymic macro phalea themselves. Enhanced •ecretioe of TDP and of a distinct mitor•ic protein /oUows the interactioo of rnacrophaRa and Immune T cells. This loter- actioo ia shown to require physical contact of Il.e two cell types and is rep.- lated by products of the I-A ro{ion of the major Ctiatocompatibi8ty complex. Belkr. D. 1., Fur, A. O. and Uwenre, E. R. Federatlon rrocerd/nst 37 (1) :91-96, 1978. OOther wrportr Na1loa.u itules of Health. From the Department of Pr tholop. Harv.rd Medical School, Boston. THE REGULATION OF LYMPHOCYTE FUNCT7ONS BY T71E MACROPHAOE In this book chaper, the author aummarires his laboratorks' work o. the role of macroph.liee i• Immune Inductios. Discussion of this work is divided into three sections: anlligen presentation /or antibody productioe and T cell aGmulalion, secretion of lymphoslimulatory molecuks, and regulatbe of T cell differentiation. In the early aatite• preaentation studic., immueo- fienicity was assayed in rlvo by measuring antibody responses in murine ro- eipienu of ayntenoie macrophalliea bearing a known amount of aeliffen, or is irradiNed mice transplanted with both macropbaRes and lymph aode ulla llrc resulla were unequivoeal: antigen aroeialed with live macrophagea was immunogenie. by definition capable of producing • strong imnwne respon... Later, studies were carried out in which amibody-torming ceNs were assayed by a plaque method. This system required the recognition of both hapleo and carrier determinants by B and T eelh, respectively. In subsequent eaperimenu it was shown that, in fac1, both hapten and carrier determinants associated with the macrophages had to be present in the same molecule. lhe secrewry function of phatocytes, which is esphxed fully in the second section of this paper, may be as important as their intracellular handling of foreign mateti.b. 73 72

65100 7193 CTR HN 012140

t In fact, both functions are deeply interceiated since the major stimulus for secretion is the process of uptake and phalloeytoaia of a partick. Mokcuks secreted by macropha6es include complement proleios, lyaoaomal eraymes, neutrd proteascs, interferon, and lysozyme. Reaults presented (n tne third sectioo here point to macropha{es as regulating a step in normal thymk maluration. In summary, macropha6es appear b ezert a le6ulatory influence al various stages in the life of the ymphoeyN - they may inAueoce the noo- aotijendriven diRerentiation of tymphocyw - as ezempliAed by the eRects on thymic differentiation; they may establii t!r fade and form of antigen to be presented or recogniud by the lymphoeyte; and they may regulate the lym- pbocyte i anti6en-driven functioea. Each of these critical regulatory steps needs to be e.plained in molecular kreu a.d b be integrated and placed in the eontest of the other regulatory /ueetiooa of lymphocytes. T?re control of mito6enic protein In an eioqueM eaarwpM of the nakcular eornpkaities and the intricate control mcchaniswr - internal and nterod - operalin6 at each step of each regulatory prooeaa. Unanrt. E. R. lnrn.rnolotkd Rtr/tw 40:227-213, 1978. Othar support: National Institutes of Healll Prom the Department of Pathology. Harvard Medical School, Boston. SUPPRESSION OF IN VITRO CYTOTOXIC RESPONSE BY MACROPIIAOPS DUE TO INDUCED AROINASE Cell-free aupernatea of mixed mouse ,pkeo kutocyte cultures sup- pressed by the additioo of escaa peritoneal macropha6a were found to be devoid of arSioioe. Wbik direct replacement of the amioo acid bad oo e8ea, the addition of glucose and 2-mercaptoelhanol to ths aolution uontainin6 ar6inine partially reversed this macroph.6e-mediated wppressioo. Complete reversal was obtained when arginlee and 6lueosa wae added to the medium and the oonadherent cells after their aeparation from tba adherent macrophasea. Aeuadiog to these data, the presence of eacessive numbers of peritooeal mac- rophages of either donor or responder type blocks the immune response of spleen eelh in culture through a depletion of ar&iee. Determin..tions of ar6inase levels showed a marked ekvation of the enzyme In macropha6ea cultured (n vitro (for 24 hours in EagleY minimum essential medium }- 10% fetal cdf aerum), in peritoneal eelb activated by prior injection of thiogly- colale, and in one spleen activated by a pa(t versus host reaction. This in- crene io enzyme synthesis by macrophases may actually be a result and a sign of their activation. The significance of these observations remains to be established, but it is possible that arginase may play an /w vivo role in the immunoauppres.ion produced by activated macrophapa. Kun6, 1. T., Broots, S. B., latway, /. P., leonard, L. L., and Tdnratt. D. W. The Jurrna/ oJ E:rrrrlmtntat Mrdklnr 146:665-672, 1977. Oth.r supportr National kience Foundation. Yawn Ila Ikpulm.nl d hl« .,.hr.l..tv nr1 IrnmunulntY, Wchb Waring Lun6 In.hauir. t 4...a..ir .-r /.d.....1.. MrJi..1 1 cnter, Iknver I THE REQUIREMENT FOR MACROPIIAdES IN THE MR'OOEN- INDUCED PROLIFERATIVE RESPONSE OP RABBIT SPLEEN CELIS It has been shown that rabbit Iymphoid ceRa an be separated ioto .ubpopulatioes that are functionally anaiotioua to B tmd T eelb of murioa systems. The present series of investigations attempts 1o determine the role of macrophatiea as necessary cells in the ilmnurqlpgk tueclioe of rabbit lymphocytes. This report deals with the interaction of lymplacylea and macro- phagea in the mitoget.-ioduced proli[erative resporre of rabbit spleen cells. Macrophaaes were removed from spleen ceN populations by plating oote plastic dishes or by passage through glass bead eolum.a. The /n vdtro proliferr tive response induced by ooncanavalin A(Coo A) and phytohemaqlutioi. (PIIA) was compared in the tanadhereM and ueaeparated spleen cell popo- Iations. The equivalent Ineorporation of 'H-thymidi.a detocted in the two populations suggested that macrophaRes an .ot required in mito6on-induced proliferative responses. Ilowevet, treatmeN of apleen cell populations with a specific antimacrophage serum during the early Inductive stage (0 to 24 hr) of mito6enic stimulalion, but eot during the Iata proliferative staRe (16 to 72 hr), results in suppression of the rppona.. Thae taulu suggest (1) that macrophates are required for the proliferative response of rabbit lymphocytes to the T cell mitot;eas, C.os A and PHA; (2) that theae cells are required early in the response and is anaM twmbers; and (3) that in attempls to depkN spleen cell populationa, methods empbyittR adherence alone are Imullicienl for the complete removal of macrophage fu.ctiow. Additional evidence for macropha6e function in the immuae response of rabbit cella will be presented In subsequent reports. Kim. K.1. ud NerscowUr, N.1. Jorrnd o/ the Rtrkr.lotwJaAefW Soclrty 2l(1) :71-66, 197t. Other arpprtt National Institutes of Health. From the Department of Microbiology. Cleotgetown University Schools of Medicine and Dentistry, Washington, D. C. MODE OF ACTION OF A SOI.UBLE IMMUNE RESPONSE SUPPRESSOR (SIRS) PRO4UCED BY CONCANAVAI.IN A-AC'('IVATED SPLEEN CELLS . .. The activation of murine Ly 2.34 T cells by milo`enic eonccntratioas of concanavalin A (Con A) leads 10 the formation of soluhlc immune response suppressor (SIRS), which suppresses plaque-fotmin6 cell (PFC) responses b hetew•bgous erythrocytes by muriee spleen cells /n vitro. The primary Iargct of SIRS activity is macrophares (M#). This wmmuniesion reports on the investigation of the mechanism(s) by which SIRS-treated Me supprew PFC responses. Etpetimenta with varying ratioa of control aupernalant- and SIRS-treated M1 show that suppression is an active process requirind at least three days for its full eapreuion, since: (1) in cultures containing SIRS- treated Mo, suppression became apparent only on the fourth day, about 24 hours after a significant decrease In DNA synthesis; (2) tesponaes of spleen cells incubated with S1RS-IresteJ Me for Ib hours and then transferred to control supernatant-ueated M• for tbe last 72 hours of culture escaped '1t

. , i suppresaion; and (3) addition of SIRS-treated MI after culture Initiation te- aulted in the delayed appearance of suppression. In addition, the data indicate that this process does not require cell to cell contact, and suggest that a second factor(s) released from the SIRStreated M• also acts to suppress the «- spotue. This particular factor may be responsible as well for the decreased DNA syntheris noted afler three days of culture and suggests the possibility that SIRS mediated suppression is ultimately due to limitation of responding 8 ulli proliferation. Tadakuma. T., and Plercr, C. W. The Journal of Inununolosy 120(2):4t1-4t6, 1978. From the Ikpartment of Pathology and Laboratory Medicine. The Jewish Ho.pital of St. louis; and the I)epartmenl of Patholory and of Microbiology and Imnmutnsbp. Washington lloivenity School of Medicine, St. I.ouis. IMMUNOI.OOICAI. DP.TY.CiION OF TUMOR PLASMINOOEN ACfIVATOR IN VITRO AND IN VIVO Plasminogen activalors can be markers of certain tseoplaams. One of these a human ovarian tumor plasminogeo activator (TA), has been found to be similar it not identical to urokinase, with respect to molecular weigh/, active ailes, and immunologic delerminants. A sensitive radioimmunoauay for uro- kinase (UK ) has now been devised and used for the detection of TA. Thia report describes the radioimmunoassay of UK with a double antibody system. Specimens of human ovarian adenocarcinomas obtained al laparotomy were used to prepare organ and ccll cultures. Sampks o( the eullure medium col- lected every fourth day were consistently flbrinolytie when tested on plasmino- ien-eonlaining human dbrin plates. No such activity was ever observed on plasminogen-(ree bovine 6brin plates. Blood samples from nude mice with ovarian tumor grafts also eoolained TA, as did blood samples obtained at lap.rolomy from vessels draining the tumors of patients with ovarian carci- nomas. On the other hand. the vascular pt.anitn{en activators in the peri- pheral blood of healthy volunteers did twt demoaNrate any activity in this maay, suggesting that they do not interfere with it. However. the radio- immunoassay ddected not only UK but alao TA administered intravenously to a heallhy volunteer. The resulls show that this radioirrununoassay can he used for detecting TA /n vivo as well as /n vltro. It is also suggested that it may prove useful in the diagnosis of neoplasia it interference with naturally occurring inhibiton can be avoided. Astedt. S. (Krl/ander, S.) In: RtKldon, R. W. (ed.): Biological Merlers of Ncoplar/a: Badc and Applied Arpcrn, New York: @IuvieNNutlh Noltand, lne., 1978. pp. Ia1-4ti9. Otftier support: Swedish Medical Research Council nd the Cancer Research Fund. Allmiinna Sjukhuset, M.Imd, Sweden. From the Research I aMaalury of the Department of (iynecolnRy arwl Oh• stctrits, the Coatul.uun 1.iKrutnty and the IsoW/.e 1.hotatory, Uni.crsdy of I und, Allmann. Siukhusct, Malmo, Sweden. I LUNG CARCINqMA-REACTIVE APfflBOD1ES ISOI.ATED FROM TUMOR TISSUES AND PLEURAL EFFUSIONS OP LUNG CANCFR PATIENTS low p11 elution techniques known to dissociate antigen-anlibody con- plexea were used to obtain inltnunotlobulin fractions from eluatr:a of lung carcinoma and from pleural eRusiow. The immueoRlobulios were uuyed against various target cells in indirect immunoAuorescence toata and ahowe Io react positively in significant titen with cells of .quamowo cell carcinomas aod adenocarcinomas of the lunj, but not with cells of normal adult and fcyl lung or of nonpulmonary lumors. ImmunoRlobulita, /imilarly dissociated from tumor eRusions of other orpoa, showed no reactivity in indirecl immuao- lluorescence tests against lung carcinoma cells. The results (rorn lung lumor tissue clulions appear promising and suggest that the disaocialion of antigen- antibody compkaea may be Ihe most efficient method (or furnishing tumor- .pecific anlibodiu. Paluch. E., and IoarA/m, N. L. lournd o/ t/k National Cancer Inelrrte 61(2 )-319-325, 1978. From the Departments of Pathology of the Leeo>t 11iq Hospital and of t6e College of Physiciatu & Surgeona of Columbia Udversily, New York. V11. Epidetniology THE INTP.RA(.'1'IONS OF SMOKING. ENVIRONMENT AND HEREDITY ANq THBIR IMPt.ICATiONS FOR DISEASE ETIOIAOY: A REPORT OF P.PIDEMIOI.OOICAL STUDIPS ON TIIE SWEnIS11 TWIN REOISTRIFS This eatensive usonotiraph describes methodological aspects of a twii research program run on the Swedish Twin Regislries, and gives a detailed account of various results from their series of questionnaire studia, clinical eaaminalions, and mortality follow-ups. Represenlative aectiom deal with: Theorelical concepts and models of the twin approach; Twin resiHrica io epidemiology; The Swedish and l). S. twin sttdies; Conceptual eon.identiotss and classification of variables auch as dose and response, ealernal eovironmenl, habib, psycho-social data, medical endpoints, and Ydidity; Hereditary aspects; Respiratory findinp; Cardiovascular findings; Morbidity; and Mortality. Sev- eral other aections deal with smoking and its environmental and hereditary interactions. These include a study of the atrue/ural and behavioral'ch.racter- nlics of thc smoker, a 14wik at the smoking related disease panurama, arw) an overview of snwkmg arrd health in the perspective of twin rescuch 76 77 i ,

:- s J r m Cederl8f, R., Frf6erg, L. and Luodmaa, T. Acte Mtdlca Sc.nlinarlce Suppl. 612:1-126, 1977. Other aupport: Swedish Medical Research Council. Folkaun /usur.oce Company, Swedish Tobacco Company, National Swedish Aaeocialion against Heart end Chest Diseases, American Medical Association Education and Re- search Foundation, U. S. Public Health Seniu, and the National Swedish Environment Protection Agency. From the Department of Environe.eetal HyRiene of the Karoliuka lnstitule, the Department of F.nvironmentd Hygiene of the Natioaal Swedish Environ- ment Protection Hoard, asd /he Deparbeeet of Medicine of the Karolinska Institute at the Scranmer Hospital. Stockholm. Sweden. T11F. FINNISH TWIN RFOIST1tY: FORMATION AND COMPILATION, UIIFSt1ONNAIRF SfUDY, 2;YOOSIIY DETERMINATION PRO('F_'DUR@S, AND RESEARCN PROORAM Tlre Finnish Twin Registry, which was established In 1974, uarled with a cohort of adult likc-seaed twin p.in, both of whose members were a)ive In 1%7. ')Ais twin population was chosen from the computerized files of the eahauaive ('enlral Population Registry on the basis of coincidence of the following criseria in two or more persons: (1) same sea: (2) same date of birth; (7) same locality of brrth; (4) same surname at birth; and (S) birth year prior to 1958 The current addreases of the twin candidates were added to the Ak, and a questionnaire was sent to all persons with a complete address in an attempt to develop a queationnaire method of determining rygosity and to obtain baseline data for longitudinal studica. The questionnaire that wat sent contained the following three questions: (1) Were you and your twin partner during childhood as alike ae "two peas in a pod" or were you of ordinary family likeness? (2) Were you .rd your twin partner so similar in appearance at school age that peopk had difficulty telling you apart7 (3) Who could tell you apart at ahoof age? 2:yttodtar determination from this question- naire was based on the combined use of the deterministic and stoch..lic methods; deterministic methods were used Ant to classify the twins as far as possible, and after that stochastic methods were applied. Verification of the questionnaire t:ygodty determination was carried out with blood markers. Overall, the Finnish Twin Registry covers a nationwide population of adult like-se.ed twins whose :ygosity status is, for the large majority, accurately known and on whom basic dala have been gathered Tbis information provides a framework for testing the hypotheaa of many studies. Kaprio, )., Sarna, S., Koakenvuo, M., aod Rentatdo, I. In: Nance, W., Allen. O. and Parisi, P. (eds.): Twin Rcicarch: Bfofory and t~oidnnioloRy. New York: Alan R. Lin, Inc., 1978. vol. 2, pp. 179 184. From the t1cpartmena of PuMic Nealah Science, University of Hclsirki, Ftcl- alnks, I'In/1/Ml 7N "fIIB FINNISH TWIN RF,0ISTRY: sASBLINB CHARACiERISTiCS. Sf?CTION 11. HISTORY OF SYMPTOMS AND Il1.NPSSES, USE OF DRUGS. PHYSICAL CHARACTERISTICS. SMOKINa, ALCOHOI., AND PHYSICAL ACTIVITY This eatensive monoRraph coweo the second portion of the baaaUr questionnaire study of the Finnish Twin Registry and deals with symptons and illnesses, use of druQs and .loohd, physical eluracteri.tia, unokin4, a.d leisure time physical activity- Aiw documented aro the b.sic aRe-seit speciec distributions of the variables tot twim considered as% Individuals, and the corresponding peinvi.e olistributioae. For most variables, the analyses cover only the responses of twin pain whose memben have both replied and who could be cla.sifled by determiniNic melhodi with reapet:t to zygosily. However, some distributions (otr pain sot dasaiAed by detersni.islic methods are also (ncluded here. Several parameten were sekcted aa Ihe variables and the basis for their choice ia e.plaieed. TM results obtained tor each variable an dir- eussed brieAy, mostly to enable eornparisoa wkA Ihe results of other twin studies and of recent studies on singktons in Finland. The methods and .ra- terials used and the distribution of psychoaoci.l variables are described ek+.- where, but they are, .everthekr, outlined in the present table of contents in order to provide an overaN view of the whole study. However, since the primary function of this partkul.r setlion is to describe the basic distributions of the available variabks, only some of the inlenelatiomhips among thew are mentioned here. Tbe others will be eaptored in separate publications. Kaprio, l., Sarea, S., Koskenvuo, M., eod Renr.r.fo, I. KwaNcrvePaletcew /uftadrr+e M 97:1-147, 1978. From the Department of Public Health Sckeoe, University of fiddakl, Hel- ainki, Fidaod. DIAGNOSIS OF'I`WIN ZYOOSlT1f BY MAILED QUESTIONNAIRE The use of populations of twim to derive epidemioloRk data hae ptovera to be most .ahNbk, aed the establi+hmcat of Ytiond Min reRiatries k,.a spurred many studies dealing with the telative hnportance of Senetk a.d environmental effects on a wide variety of human attributes. Ilowever, wbila recent developments in NaliNicd and date ptooerlnR facilities have .rirted the progress of lar4e-acak twin atudie., they have also created new melhodo- logkal probkms. Among /hese, is the need to develop techniques which are sutriciently reliable, efficient and generally applicabk, so (hat the resulla of epidenab4ic studies are fntdmtionally comparable. One of the Indis- pensable requirements is the ability to differentiate accurately between the monozygotic (MZ) and disy{olk (DZ) twins. le recent year., both a quea- tionnaire and the use of blood lesls to determine pdymoryhic blood reuken have been used, but neither has been found compkkly salisfac/ory. Ilero, t1r authors describe a further development of the questionnaire method of sygosity diagnosis, and compare it with the results of blood tests on a smatl sample of Iwins in the Finnish Twin Registry. TAe method was based on the answers 79

K) #-i of both memben of a twin pair to two questione oe aimilarit7 and eanfu.icn In childhood. 'i7re algorithm of the method wae used to determine the zy6asity etatus of a twin pair at two di[fereat levels of certainty. Blood mark•:r deter- minations of 11 polymorpbic marker syeteer from a aampk of I D4 p.irs drawn at randorn from twin p.in elusi6ed by the questionnaire (93.3% of all resspondent pairs) were used to test the valWity of the method. The yuestion- esire and blood marker diapno.is were In complete agreement (1009G ), but the stricter kvel of certainty kft 0.7% In the oooclassified Sroup. The umpk'a genetic repreaentativePe~a waa tested by the allele distribution of the marken as compared to the )~rnoiab population data, and by the distribution of the number of intrapeir dUfercncee lo blood aeerkera S.rna. S. tr d. ( R.nrar.fo. I. ) Human Heredity 2s:241-214, 1978. Prom the Department of Public Health Sciesoe, Univenity of Ilelsioki, and the Ikpartment of Immunobiolop. Central Public Health L.boratory, ilelrinki, Finland. CIOARt?TT8 SMOKING IIAB(iS AND URINE CHARACTERISiICS 7be relationships of cigarette smoking status to the pH of voided urine and to the abnormal presence of protein, glueoee, blood and urea were studied in 33,000 while and 9.300 black men and women who participated In multi- phasie health checkups al Kaiser-Permanente medical facilities between 1964 and 196d. Proteinuria was more common in rrwken than in nonsmokers Rprdleaa of race and sea and more frequent among heavy smoken than light smokers. Only one of seven different variables, ak.ohol consumption habits, seemed partly interrelated with the amokin6-proteinuria arociatioo. Proteinuria was also mon common in smokers who were eondrinken or light drinken. Tbie situation was reversed among the beavier drinkers, however, with the soeemoken showing proteinuria somewhat eswn frequenlly. Cessation of smoking was awt accompanied by a relatively bwor proteiouria prevalence. Furtbermore, the preaearoe of protein is the uriae of emokers did not appear to be indicative of moro serious renal disease. 7b urine Rluco.e raponse to oral glucose challenge was different in emoken and .oo.eakera, probably due to the higher average ooe-hour serum R/ucoee values found In smokers. AI- thwrgh the mechanism behind this difference is unknowe, it seemed related to alcohol consumption. Hematuria, bacteriuria, and low pH tended to be more prevalent among smoker., but these relationships were sot comistently ai6- .iAe.et. D.ks, L. 0., Frirdrnew, Q. D., Skilelaub, A. S., Seltzer, C. C., and Ury, H. K. Nephron 20(3):162-170, 1978. Ot/irr arr'ortr U. S. Public Health Service and the Kaiser Foundation Re- search Institute Pr4.m the lkp.nnwm o/ Med.aal Merbods Research, Permanente Medical (Jruup, l)s\IsnJ, ( •I 80 l t PSYCHIATRIC SYMPTOM DIMENSIONS IN T71B CORNELL MEDICAL INDEX AMONG NORMAL ADULT MALES Items from the peycbiatric pwtlon of the Cornell Medical lndea (CMI) were subjected to factor andysis In order to determine the empirical sryropwo dimensions in a sample of 1.682 normal adult makts. Tbe d: faclon tbu were e:tracted were interireled r IrrilabiNq, Inability to Cope. Depreeelo., 7lmidiry, Normal Arniety, and Clinical Anxiety. 7bese corresponded oornewbat to the clinical groupings in the CMI, as well .n to other facton discovered iN other studies. The consistency of these results does tat support the b7Potbesls that the CMI M-R sections meawrs only 2 witary factor of .eurotie coas- plaininp, in spite of the fact that they ap showed small poeitive correlations with a measure of Neuroticf.m. 'Ibe authors recommend some rnodilkatloai of the CMI that could enhance its utility r a diagnostic or perswnality Mrtru- mest. The deflnitioa of a siew section to inchde the items In the llmidity factor, and the addition to the auestiooeain of certain items indicative of neurotic obsessions and coanpulslo.a, would be pertkularly important. Cosre, P. T., !r. and MoCrae, R. R. Jorrwaf o/ Cllnkd hyc•Aolop 3l(4) :941-946, 1977. Otitier eurprt: Medical Reseateh Service of tba Veterans Adminiatratio.. Prom the Normative Aging Study. Veterans Administration Outpatient Clisk; and the Department of Pqcho{op. CoUep 1. University of Massachusetts at BoeWt.. OBIBCTIVB PERSONALITY ASSESSM6Nf' 'ibis book chapter triee a draw a bta.d, empiricaily-beaed eketcb of eonnal personality and ib eha.p throughout adulthood. It abo attempts b make specific links between normal personality aed clinical gerontology. 'I1w are three sections of the chapter qhich are addressed to: (1) longitudinal d.u on stability and change in normal personality; (2) data on the devclopore.t ud validation of u IaHrumeet /o meawre .e.er.l facete of opennesr b ea- perience (a broad domain of personality which haa important Lnplieatloae for questions of personality developmeee, weM-being, aoq the quality of lire); and (3) empirical data on a"new" condition of interest to cliniciane, the "mid- Yfe crisis." The evidence cited here shows that obiective, and in particdar. V f-report methode can make signlAcaM eootributiona lo an understanding of ult personality. Objectively meawred personality trail.--at kaN the broad dimemion. of amiery. opennas,.and eatravenion-are meaningful and stable dimensions of Individual diQereatee whicb can be merured by standard te.u applicable to adults through the eighth decade. Perhaps the major conclusioe to be drawn about objectively measured peraondity, at least In men, is that It remains remarkably stable throughout adulthood. There appear to be decliese in a few variabke--eatravenion and masculinitf. for esample---0ut theee changes ate very small In comparison to the range of variation among Individ- uals in any cohort. In addilion, according to these data, most men do not go through a`mid life erisis" at all; those who do, do so at no particular a3e; el

i and the crisis itself may be rsotbia8 more than a manifeatation of loog-.tand- in8 inatability or oeurolicirm. Cosra, P. T.. Jr. and McCrae, R. R. In: Storandt, M.. Siegler. 1. C. and Ella., M. F. (eds.): The Cf/nicd Piy- cAdory o/ Aging. New York: Plenum Publishing Corp.. 1978, pp. 119-141. O/Aer supports Medical Research Servioe of Ily Veteram Administration. Frorn the Normative Aging Study, Vetena. Administration Outpatient Cliuic, nnd the Univerwtr of Massachusetts M douoa I . Active Projects ANXIETY. EXTRAVERSION AND SMOK.INO In earlier attempts to understand Us wativ.tiooa underlying cigarette omokift, many etudiea have saamised tM re4tioa.Npe betwcea cl:an:tle arnok- (oe and anakty. and eat.avereios. Sonse Msdks have fouad as aa.ociNion with eatravenion, some with anaiety. The present os, however, w.a b..ed on the hypothesis that smokers would be both nwr+e euUra.erled and mwv aaitious then noe.moken. Subjects for this study wsrs draws from ttse 2.300 adult mak volunteers who are participants is the Normative Aging Study, and anxiety and satraverdoa ecora were etamioed Is Rroup of men srho never rraked, former li8ht, former heavy, current li8ht, aed current heavy smokers. Two bxfepeodent rneawrea of both uaita were used. Heavy current .nsokers were .cen to be aignifkantly higher than nonsmokers on both mwasures of oeuroticiam or anaiety; there were no differeeoca between groups in e.tra- versioq. What 6ttk evidence was eeen here for more smoking among e.tra- •erta appeared to be an artifact of age: hrnh ealravenron and cigaretle amok- ies being more characteristic of younger men llse small magnitude of the eorrelations reporied here suggest that fuwre research must become snore speeifie, both in terms of a more meaningful claaarfscation of .nwkere and in terms of more apecifle facets of personality. MeCsae, R. R., Corta, P. T., Jr. .ad Boe.l, R. IritirA Joornd o/ Socid dtd CHnk./ PrycAolop 17:269-273, 1978. OtAer eupporlc Medical Re.earch Service of the Vderao. Administration. Frorn the Veterane Adrniniatratioe Outpatient Clinic and the University of Massachusetts at Bostoo. v Following h a Uat of ehe principal inve.ti6ators, or lnatltudooa, od projects under way or activated in the period since the. previous Report, together with the rtapective project titka. Completed projects are listed la a later.eetion. PRINCIPAL INYIXTIOATOR OR WS7TiUi7ON LPd) O. AsO(M), Pr D, Pr%aror o/ I1ocArwdury and lyd/n RewrcCt. GN- 1er for Staie Reecarcit. The U.i..reMy of RocdeMer Medkal GM.r, Rochw ter, N. Y. MARIO D. ACE70, Pw D., As.ariare Pro «.or o .P ~.~. Medicd weYla UOieeflnL. V CAS~ ieb.rooe. R )OSEPH C. ARCOS DSc, Projraaor of Medkanr. 7101a.e 1lai.ereity School of Medicine. New Orleans. ALAN K. ARMITAOt3, Prr.D. Re.rreA Director, Harkto. Laboratories t?r- rope, H.rroprs, North Yarkst.ire, Ery- land. LPSt.IE taAER, M.D., A.roNarr Pro/rrr sor oJ Medkiet, CdwwAia Usl.ereity College of Physicians A Suraeon~ New York. YVIt.I.IAM P. tiENEDICT, M.D., Aas/rr- enr Professor of Pediatrira. Uwiversitr of Southern California School of Medr- eine. Division of Hematolof y and Med- kd Oenetica, Crildrew e Hospital of Los Angeles. 1.oe Anacks. DHA 1. van den taPRO, M D.. RerracA rrdiarrkkrw, A/Iwhrr rrofersor /w Alo• rtaNak•r. Ueiversity of Csli/%nis Schoo/ of Publle Heahtt O.Llasd. RICHARD J. BINO, M D., P<roler>a of Mrdkine. University of Southern Cali- fornia School of Medkine, Loe An- tcks; YbiNnr Auoclare in Dionrtdr.al lnrinrrrlnf, Cahfwnia lnuirule of Tccbnu/otr; Dnruw of Crdiolop and Inrramaral Mrdniwe, Ilunlinaton Menwrid Ilospilal, Pasadena, Cal. PROJtCT iTil.ti Relavioral ase b{odrkel ansdiee .W a lcori.. 3laroqeci/k binding of .lectire Activating wetaboWrs of dlaNyt ad cyclic .itroe.uninee a.d 8e.etlcs of tko MW en:rnw Inducen. Relatloo. ~ydrocarbo.-.Irroesraw eyf Suoeal ab.orpioe of labelled .Ioallr by the anaestt,etieed cat from daar swoke u compared to that from cl8areua Ciantle .nokiaa Iw .orsnotead.e .d typerren.i.e wbiect.: blood preawe, rcniq, aldoeterooe aad catarJolawi.e reya.w MJianasN Iran.formatioo, mwaReweelr and Rbrindlnita productiow of citaette rwoke cawkn.ae fractiora Malipaat uu.fonnatloq wwt.p.aeY end pornder activity of cigarette roke condcwale fractio.u Pulrnos.arP fundlon reere in a&Ae.oeele and their parenr.--aa eoldemiooogk aopooch In6itsitios of ehOleMerol uptake by 7- ketocltoieqerd FRec/ of cerbon monoxide qrl cAolererd meaabuliun of Ihe artcriel wall 8) 82

PRINCIPAL INV[Ji1CATOR rRO)[CT Tyili I rRINC1rAL INVl3i1QATOR rRQ/RCi 7ti1s OR IN11'1TVTION OR INSITIVf7ON FRAN~OIS M. SOOYSP *N.D., Auocl- arr rofrrror o/ eiorhrmiury, Rusb- PresbylerianS/. Luke'e Medical Center, Cricago. RAYMOND IIOSS& M D., Aa.odara Dirrctor, No.mat/ve Aging Srudr, Vet- araes Adminialratio. Outpatient Clinic. Boaton. A. SONIA sULST, M.D. Ar.ot/au t•n.- Liwr of Mrli.lar and f hyr/oloty, niversily of Oregon Heellk Sckaoce Center. Portland. P.LROY T. CANTRP.L.L., Pw D., Chalr• •.an, Department of Pharmacology. Teaaa College of Oueopalhic Medicine, North Teaae Stata University. Deno.. ALBERT CASTRO. rlr D., Dkrcro.. Hor- wrone Rr.rarch L.Ao.atory; rra/ruar of rharmacoloey. University of Miami School of Med/ci/le, Miami. Fla. JACK CHALON. M D, Auociaur tro/rs- aor o/ Anerrhrrblot; y. New York Uni- renity Medical Cenler, New York, FRANCIS C. CHAO, M D, M.D., Srn- lor Inrert/gator, Centcr for llood R. .eareR Soalos. CHARLES O. COCHRANP M.D, Mem- 1er, Dejearrnwwro/ Immunnpthdogy, Scripps Clink and Reseaeh Founds- lio4 La lollti Cal. ALLEN B. COHP.N, M D., Pit D.. A.- aocia/e rro%..or of MedG lae, Chk/, P.dmowary SecNow- Temple Uwversity Scienoea (:eMer, P\iladclphia. ALLAN C. COLLINS• M.D., Auociate rro/ra.or of r/urn..co/ogt'. Indilutc for Behavioral (iendic., University of Colorado. Boulder. rAlli. T. COSTA. Ir. Pn D.. Associate f'Y../nror of hyrhnh.ty. University of Ma..achuselts at Su+tnn, Dorchesrer. IRVIN(7 P( NAW/IIMI) M/1. 1•r../r.. n.. ...J . h... ,..n f..f.......r... ..r .li .r. /. . 1 ~. ......,. ..r t ... . /1. ~r . f \/..1. ... 1 .. ... In viro .rad In vlrro rearoneea of endo- tbelial oells and platelets to nicotine and eataaa frorn seandardired cigarette .moke oo.dee..tea /n .iw ud In vitro reWon.d of endo- 11cIW oeUs and the vessel wall to eico- 11.e A wokln` research poject in the Nor- asNlv. Aginj Study The rola of alphta-l-anlNrypwn delkkney aa a risk factoe In t!e development of chronic .Irwaya oMtruqion Aryl hydrocarbo. hydronyla.e In single oelle and sLbropulationa of humnn lyrn- phucyteS and other cella Niootina 1s blood: detedion by redio- immunoaea.y Epidemiology of traeheolxoachial epilAc- lial multiaudea/ion Platelet activalion and blood hypercoallu- IabilGy The mediation of inAammatory iu/ury, of tissue The cherninl basie of Ihe design of lhera- PeWk .Renu Lor aelirroteolyuc actit• Ny in the IreatnxM of emphy.ear Oenelk analysis of neurnchemkal and 1Aa haviord efiectu of nicoliea and alcohol Tlte relations between amoking motivcs, personality and feelinp I oulncyle e1H/aK c(Mn rmrnl inleuc. .~.o.r in the rl.nh.sr u emphysrm• DAVID W. CRUMrACKER, rN.D., ho• /nwr and Chairman Department o/ Env4onmrntal, roIrtulon and Or~f w/rmk siolop. University of Colorado, Boulder. WAL.TP.R B. ESSMAN, M D., Pn.D, Professor of Psychology anf e/ochem- btry. Qutens College of tle City Uni- versity of New York. Flushing. WII l1AM N. PISIIMAN, rar.D., rre.l- drnt, 1.a ldla ('aaeer Re.earc\ Pounda- Iiuq L.a /olla, Cd. IUDITH ANN POSiP.R, rM.D., Aaacl- afe rro/e,ror of slocAemlrary. Univer- aily of OeorRia. AtAen.. AARON E. FREEMAN. hr.D., SckntlLt, 1 a/dla Cancer Research Poundatiol4 La Jolla. Cal. LARS PR1slRO, M.D, Pro/eisor anI Chairn.an. Drpartnsenr o/ Ew.bow- mennl H)~lenr, T\a Karoliaaka loetl- lule, S(odblm. GARY D. FRIEDMAN, M.D. An A(iatawf D/rrctor, Drpr tment o R/rler/ etA- od. Reararch• K.i.er ~oundatiow Re- aarc! InMitwe, 041404 Cal. MICHAEL O. OPOKAS, M.D.. rw.D., rro/essor and vke-CWrmaw, D mento/ Mrdklnr, Univeraily of Cali- lureia School of Medici.e, Davis. TERESA OPSSNP.R, M.D.. Aasxlatr Canerr RrarrrA Srknti.l, Health Re- Karch Inc., Rosweq Park Division, ltuf- fdo, tl. Y. JACQUES E. OICLEN, PM.D.. Assistant Pro/rtun, [.Sorarory of Meliral l'hrmbtry, To.kalosy and Hysknr In.titule of ratholoay. Universily of I iqe, I.kge, Relgium. R(INAI D W. (Ill.1.P.T1E• Pu.D., Dkrr- tor, suir S.kncr Rrwrch Unit. Can- cer Center of Ha.raii• University of Il.wall a/ Manoa, Ilondulu. OP.RAI D/. OLP.K'll, M D, Co..sdtant in Mrdi.lwr, Reeearch laboralory for • Alkr{ic I)uea.ee, Maro ('link and F.wndation; Auoc u/r /rn/ruor o~ In- frrnal Afrdi.inr and lmmuaol.r~r, Mayo Medicd Sctx.ol, Rochesler, Minn. Uoetk .eA eavko.rne.lal /aaors d- Lecliy rrnokiy belavior Correlala of Mrerr-tob.orn aa.oka I.Mr- adk. M.rke. rro/lla In kroecaid eooars ad necrlas,." of a.rokera aed .o....okara Lvolvrsetl o( d.wle fibera I. h+.d dlrw Rodent and Munas luy ep11he1W odl oyi- 1we r a tool Lor careiooRene.la reaaaeJ Iw rlao !!ridemiologkal .tudiea on the S.e" twin registries Longitudin.l e.rnoker study and t6a oo- bon morul'ay study CirculalinR p.wcreatk dawaee ad a.- Phy.ems iw man Pk.rt.aooReepka of ooaMyalio.w nd kpwg cancer ri.k CiprHte amoke and polycydk "o- tarbos aetaholiam Is lung .nd kldeay P-/feet of tobacco Oyrroducte and other envirorrnentd coMun/nanta oa ly.- pboid atl homing and fundio. Human hypersens/tivNy lo antigeea fro. 1uA.cco 85

~ ~ PRINCtPAL INVFdi1GATOR PROJCCT TTil.t OR INST17U170N , 1 JOHN W. (SORROD, D C.C., Lecfwer Iw /ioPAscmocl, Chelsea CoUege, Uru.ec- To.kolo" Mudk• on 'Pyridine-Noa- ide•' and •oare related onmpounde sily of l.ondon, Losdo•. IIODA A. OUIROIS, Prt D., A»oclae AHII •nd related immanocbemk•1 muk- : Prof.tnor q/ Prrrewllre Afedicinc and PrAlrc IfealrA, Creighton Universily School of Medicine, Omeh.. (Now A} socwe Pr../cssor of Commrwify and Enrieonmewrd ifeficine U iv itl f en: a wooeptibfl'ny indea for lung pscer , n cn r o California College of Medicine, Irvw.) CAROI INP S. IIAI.I, M D, Ae.oci.ri Prof.uor of PrNurks and M.Iicby, Universily of Rocheaer School of Med- The rel•(io..fl of lower respirelo.y tract 'dise•ee iw Mlency and the development of chronic h.ns dlse•ee In .dults: devel- 1 kine, Rocherer, N. Y. oPwrc~1.1• oourr of phywdqic bwae• I INDA M. HALL, Ptr D, Assbtawr Pio. /ctwr of Qiolotl, MewchusNls (wMi- lule of Teclnolotr, C•mbridge. MARCIT NAMOSH. Pu D. RncrcA .Ltsori.re, Dry.rrmrnr of PArtiolotl and SlopAyrks, OcorVelown llnivcr• si11• Schools of Medicrne and Denlis- try, W..lin)to4k D C. PAt)l. HAMOSH. M D, Au.wi.re Pro- /essoe of PAls/oloer and RrooArucs, ond Metrnnc. Cseorjelown l)nrversiq ScAoole of Medicine and Demiury, Washington. D.C. NORMAN W. HPIMSi-RA, Pp D, Pro- /nsoe of PsycAoEc.Pr: Director. Hum.n Farrors L.bor.ro.y, llniversilr of Soutil D•Iot•, Vermillion. Oe.etk diRerenoe. ka akotine eensi(ivitr in DroaorMb wv4wotwrer str•irr Pain.t•1 luy de.dopne.t, rner•boilnn •sd eit•retN smoke PRca of emokinR o0 evolution of tAe m•almuss espir•tory Sow-volume carve Behavioral eflecls on nonsmokerr of ea- Porure to emoking Son+e Eeh•.iote0 •+qects of smokinR and emok/ng depri.•lioa C.AR(N. 1. HP.NRY. PN D, Director. Dc- ~.vImcM of Et/crimcwtd Onrology, Micrubiotogic•1 Aswci•tes. Inc.. Se- thcaJa, Md. HNRRFRT S. HP.RSCOWITZ, PND, Arur.idre Pro/eswr of Afirro6luioly, (h.wgeluwn Univer.ity Schools of Med- kine and Dentirtr. Washington. D C. RONAI D R. /1UTCHINSON, PH D, Re- sear. A ItIII. roe, FonnJ.IM.n fIM Nch•v- ior.l Reseerch, Auaus4, Mich. At PININSP I INC)FNITO, Pu 1). Ar.o- .ia/N Pr../rr.,v ,r/ Ylr..rrnr..•t.,Kr. 1'at ('.rulm• Ilw.u.r1Y %.tnMJ uf fllydi. clnr. l)rctn.dlr, N (' E!vIu•liow and c6•r•cteririlion of an •1- kdine elulloo •wf u• measure of pul- mon•ry DNA damage induced tr chem- ical cudnoRen• of Ihe chemicals in ciduelle smok• P11eds of cigarette snwke e.posure on develop.nenl•1, ulluler and molecul.r aspects of tbc Immune response POects of •cvte and chronic nicotine •d- minis(r•tion uPnn •Rgressinn end M.ad pressure re•dwns in r•rs and humans Actions of urAon nwnosnk and tobacco reuke on rethn•) mtt.bulum .rrl (unc- /ion 86 I eRINCIlAl. wv1<itSOATOR OR INd1liUfflON HARRY L. IOACHIM, M.D, Anend/wt Pathologist. Leraa Hitl Ho.Piuf; C c.f Pro/enor of PaAobtl,~~M ! Uwveee.My Cdkp of h Su.leonti New York. AARON )ANOPP Pt1A, lro/tuae ol P.rAoIoPT, He•rlh Sdoeoe• Ce.ter, Stue Uacver.Rr of New York at Stony Rrook,S(a.l Brook. WILLIAM l. JUSKO. P(tD, A..oc1.r. Professor of PA.rm.cewdct DLeaor, Clinlcd PArnrcol/wtrke L.bor.«ory, Mill•rd Fdlswoe• Hoepitr, RrIR•lo. ESDWARD L. KLAIRkR, M.D., Senior SclewNu, The Wo.ceater PoundMiow for P.aperirneMd Riolop, Inc. Sfrew• bury. M•r. )EROMH KLEqNeRMAN, M.D, Neod, Dro.rrw.ewr of Pahofop and l.rAof- X Resercll, S.iet Lsde'e .el•od. (Now Peo(es.ar .n/ wr.w DeNfn.rwr o/ r.rAobsl, Mowt Sct.ooi of MedkJ.e, New Yerk.) KLAUS 6. KUCTINER A(D. ho/eaa. o/ Iiar4wrWr1 Iw Orr"lki. Dy.rt- rwewr of dbr/km(atry. RwA Colkee of He•Rtt Sciences and Rsri Medical Col- klie, Rwb-Presby/ec1..-3t. Lrte'e Med- icd Cerer, Chicago. SilO KULLANDER. MD. h%>1.o. and CA./rw..n, Der.rM.rwt of Oitrer- eJc, .wt clnecofop. Ud.enily of Luod, Lund, S.rede.. ARPL LAITIIA, P(r.D.. DMecroe, New York State Research Inaitute for Nt+r- rochemislry and Drug Addidioq New York. )()SFPH M. LAIIWP.RYNS, M D., Phr 1). P+o/csto. (Ardinir/rs and CA.Ir• wvw, D./urtmcnr of PNAofog1 and M.croscoplc An.romy, t!aPerimenl•) Labaetory of Pulmon•rr Hlaop•/bl- ofty, K•lholkke Uwivereileil t• Leuvee, Leuvtn, Selaiua. JAY A. LP-VY, M D., Asdu.nr Clinical Professor of Medicine: Reu.reA Auo- r/or., (-.nar Research Institute. l)ni- versity of ('.ILforni• School of Medi- cint. San t'r.nci.co. rteo~tR-t zntt Tie ior.w t.Woo.• L LaR ardwrr Supqseeslow of Protc•w Wlbi(loo by dR- •retM uswk• Pdod of rrokloy and Me oerWbe e. drug dMPo.itio. Bf•W of .Ioo11r l.ptbm or lek.•.lot ..l Idoo/ pnarws i. rW Purtier ..dwum of .do" l..p of f•sil!/c e.rpiP•an• llsw Ero. Mkas MokoRic.l ArocWe• DistrRuMlon .nd rs.pn.. ot •.M• Islas- wr•sr sqt.ke •ed decar0oaylNkor ka. (*l/E. 1' .1 i.~aaassil n.edi• ~ Lee.t eegrl•tiow of .or.n•1 n.A Puk.okoRke h.nyio. LAre.om ot osokiy os k.s•s foeW /rowlh •wd /oMw•ul and on RtRi.olrsie /n tb lloed 01 Ptean•M women. AowrwnWioe of oieoli.• •rd/or danye to kmn•s p1•- oeMd ned foel•1 lusy tiswre• T1e eRecA of .ka1Me ..d c•rbow nw- oald• o. the tr•n.Port of •wd•o •clds kelo Sr•in, •.d on Prouls rnet•bolies The oturoePit6ellel bodies: lrelr rok. and Mruc(ure •s IMr•Pulrwowrr .aro- (chewrokecePtors in normel and v.ri- ous physiological. ph•rm•colodic.l .nd P•Ikologic•1 condilion• Possibk gcnetie determin.n(s of clesksl e•rd"eneds f37

PRINCIPAL 1NVETflCATOR OR tN1TTTtfI1ON HENRY T. LYNCH. M.D.. !ro/rasor and CAainnan. Drpartmrnr of Prerea- tire Mediclne and rrblic HrolrA, Creighlon University School of Medi- eine, Omeha. Neb. PRANK ARTHUR MANNINO. M.D Au/sranr Professor of Obrrerrlcs aWJ cynecolocy. Women: Howel. Los Angeles County/Ilniversity ol .SoutAerw California Medical ('enmer, I.ae Angeles. R. RUSSPI.1. MARTIN, M D., rrofiasor of Mrdklnr, awd Microbiolnfr • la.- '^unoioly. S.ylor ('olkbe of Mediclae. I louston. RP.OINALD O. MASON. MD., PwD, 1 aAologbrln-Clilef, Memoriel Hapi- ed, P.wluckel, R. 1. (Now Professor aad CAairman. Deparrnant of rarA- olory. Univer.ity of Sou1s Florida Col- kge of Medicine, Tarnpa.) OlRALD F. Mc(1P.ARN. PwD. Dl- recto., 1nsNrare for Arlbrio..l Ce- artlcr, Drpartn.rnr of !'q tAoloRy, Ilnl- versily of Colorado Sc6cwl of Phar- mecy. Soulder. HPRRP.RT McKPNNIS, )r . Pro f)_ ho- /errof of tAa.nrarol..Ry, Medical Col- ~Re of Virginia, Vir('ommon- .ealt\ Univerdty, RicMror~d. IIANS MFIFR. D.V.M. Senio. SaLl Srl- enrlu. TLe Jackson Laiwratory, S.r Harhor, Me. MICROSIOI.OOICAL ASSOCIATES. INC, dethc.da, Md. l. ANDRPW MfTC.HNI.I., Pro O, Arrlsr- awt rrofrrrrsr of Anatomy. Wayne Slale Itni.erury Schwd nf Medicine. tktroit. PRO1[CT TTIli Statiaic.l-lcnelk ev.luatton of risk fac- ton In lunb cancer PM.1 and maternal effects of cilarclte trnokin/, nicotine In/ection and carbon twonoaide inhdation in the pregnant RAesrs nwnkey model laduclloa of aryl hydrocarbon hydosylase In lymphocytes and pulmonary macro- rhas- PJleeta of wwokinR .nd nicotine on growth .nd fuactiow ot luman endotAclial ulle Heredily and tobaeeo-relaled behavior in t6e nwuae Cooperative studies aimed at the develop- ment of immuno.seays for nicotine and nicotine rnetaholi/es OncoRenesi. In the rabbit: genetic suxep- lihility, vertical trensmiaion of virus and envlronmeMal influences Tlanfpl.eenlal effects of nitroocom- Pounds Deve/opteM of a mouse model system for senelk wrscepihilily and its relation- aGiP 1o iw riro lung eareinogenesis Smok. inhalation earcinogenesis euedks In aska Human AHH sludke Research arvices in support of another prokeY (see Levy. Jay A.) Nicollne-Induced altertllons in Nastncyd deve)opment. imPlan/Nion ami uterine runction in the rer Imp(entetion and uterine funclion in the ret m 1 PRINCIPAL 1NV1a17('ATOtt OR INA1TIRfI1ON FP.RID MURAD, M.D., Pt..D, Director, Division of Clinical rlurmocelody, DeyarvneM of latesad Medicine. Unl- venity of Virginia School of Medidee, Clarlottc.vilk. OP.ORO B. NL'URATII, PN.D., ldkro- awdyticd Labo.arary, HambuR, West Ourwany. FRANZ OPSCH, Pn.D, Professor of rh.rawcologr: Head. Section on Bar- cAeraical PA..macology, Uaiversily of Main; Mein; West Oermay KPNNPTII PFAIUPN ht.D., DMtdor, Drpartmeat o/ laalecd« /loloq, Iledth Researe4 lnc., RosweY Park Division, Buffalo. CARL W. PIERCI; M.D., Pn.D, Profta- ior of Pathology and 1'fkroiidory-lar- arr.wofosy. Wa.6lapoa Uuivereiq School of Medicine; l.r4oloR/u-Ia- CAir/. 'IU Jewish Hosqilal of S1. L.owds, St. I.oui.. ILARI RANTASALO, M.D, l~easor and C1lairaura. Dep..rnua/ rblk Health Science. University of Helsinki. Helsinki, Finlaad. MARILYN A. RASCO. Pw.D., Assistant Siologiat and Asa/s/aaf t•ro%aw+ of Bi- ology. The University of Tew Srstem Cancer Center. M.D. Anderson flos0i- ta) and Tumor I.e1NuU. Houtloa RONALD E. RASMUSSP.N, Pa.D., As- sociare Adjunct Professor M Coasa.r- aity and Environmental Medkine, Un1- versiq of California College of Med4 eiae, lrvine. LYNNC M. REID. M.D., M'olbacA rro- lrsrow of ratlrology. Iluvad Medicd Schod, Boston; Chairman. Departraent of Parholoty. Ckildre. i HosPitd Medical ('eMer, Boston. HI'.RRPRT Y. RPYNOI DS, M.D.. Ar- .oria/e Professor of /w/rrnal Mrdkine: Heod, t dmonarr Section. Yak Univer- aty School of Medicine. New Ilavea, Conn. )011N A R(1S1?('RANS. PN D., Asw- rrarr trofruw of rAanmac.dogl. Medi• ed ('dlete of Virginia. Virlinu Com- monwealls University, Richmond. TRck1Lt.'i 1111= Mecr.nirn of nitric oiide adivndoa af Rolaylatc cydue Nitroernl.es itn tob.4oo ..d iu assoka: oocawre.a, forsutiow a.d verter Trw/ormatioo of evk.ryolk oells by ttolycycGe Rydrowswu occurring in ciprette..rota: diacriasieatioa betww <•. Iruporta.os of v.rloue cayonee A ~c tle leet of t{lucurodd.es in kl.d- der ut.oer Biology of eupprtaeor T celle Til. Plani.Y 'iT.ia Registry EPidernlolosical Mudies of 1M PYt.ira Tbi. Rqwtrr Hydrocar0oa wtetaboli:ly e.eyww tod luq c..oer A ge.etk M.d~ of DNA rq.k ks I~Dr.~ Mraim of sia T1e eReets of irritatiort aad drup oa drvay ePilheliusw - .a .aPerloeWl study of wrocMairw., Pss.+dio. r/ arre Rroncb.lvcdar LvaRe iluN ia pln.oa.ry e.rcinana: secraory coatloaeal af 1.- awnoffluhulia A aa a mark.r of t.eo- pl.stk growth Discrlmintllve stimulus peopeales af dos- tase and related co.apw.ds e9

tRINCIf AL INVLli1(`ATOR OR INSTiiUT1ON UNA S. RYAN. M D., Srnlor Sclentlu, I.Penkoleou Cancer Re.cercit InMI- lute, MLnal• AtsocWe rro/euo.ef Sltdiclnr. U.i.erdtr of Ml.mi Scfod of Medkine. Mier.` Fle. •. V. RAMA SASTRY, D.Sc.. M.D, /roJrr..or e/ rArwrocologf. Vewder- lill Uairenity Sclool of Medicine, NuAvilk, Tcr. lAKOB SCHMfDT, MD, Pr.D, A.- darrV rio/ca.am of liocAcwrlMry. Di- rlrlow of ! kef Sckwcer. S1e1e Ud.er.My of ew York M 3/0.y .root. Sw.r Isroot. CARL C. SP.LTZER, Iw D.. Hene.ery Rrrrrctr Awrire, 1 eebadl Mu.euwti Harvard Uniren/1r. Cunlride.. Mer. OERALD SHKLAR, DD, CArf.. A. f7rorlrn Ire/e%.er of Or.~ 1N~~ >'; Heed, Dr~r nncwt of Ore/ Mrdklne end Oral raAolop. Herv.rd School of Dea.l Medicl.e, bra.. NATHAN H. SLOANI; rtr D Iro/ea .ar of lloclbwdrrry, The UnlverYlr of Teneeuee Centa for tbe tleeir` ScknuS McrnOAle. .n cAe., .ncAe P. LOUIS A. SOLOFF M.D If Gr DWIw e4Aej Serrke Pro/eaiwt froTerwr e Shdkiwr Dkerro., Re- wrrA LI L.~or.ror~, Tnnple Ud- ..reM Heeltb Sciences Ce.ter, Ihile- dei~e. THOMAS P. SP06SP.L, M.D. CAfe/, Metir.f onc , Un4, Mu..clueNte Oenerr doMOw. DAVID W. TALMAOE. M.D.. Director, fYet.S-IYerlnt Lrrt Iwnltra, U.Ler.i1y of Co(ar.do Medical CeMer, Deaver. ANDRF.W M. TOMFTSKO, !1t D l.e.- Ilrwr and Dbecav of RerrrcA, litro. I.eEor.torieti L1d, Rocle.tet, N.Y. JAMES TRAVIS, h1 D Pro/eiaor of RbrAemlM~, e Z).iveniy of t7tor~iti ArtAer.11 OPRAI D M 1URIN(1 M 1), f•o/.....r o/ Mrdulnr- C.hrwhe Ue..crwy (.4 kse of 1l/ea4we • Surleoea Nc. York. fROJiCi 117112 M.tabolk ect).Ftiee at Ittl.weery endo lieliu. Idwea of tacotine ow Ib rek..e of noe111cRoliee h tb Lwnew placenta and Ye Iop/kallone o. the t growth Ce.trel .lootl.k reoepore Co.rltulio..l ettdiee reletive to smoking e.d coro..ry leart dieew ~ d.opr.i~ viuwl. A eed te(- Munmdlan Wai enzyme: Aloydroapnttbllbcnr.o(e)P)rene eyn- thetear.---Sioclemical and biological rudiee The role of IacA6is cbk.tnol ecyttrene- terer (LCAT) (n cbkelerol uwport lo end fraw perioAerel Iiaun and its modifkalios by smoking ..cropVN y of w pulmonary Tie roM of ttuaoo~aae-ledueed f.ctore 1n easoer i.rat,nhr 1'rolind nicotine receptor eilee faloclRrnMry of chronk obbtructive lung di.eaue 1'releolltk eaeya+ee and in6iEilore Is cm- phy.eme (linnkd basis of ti.we deurucrbe In oletrutlive long dra.ae. lRINCRAL INVE,Si1GATOR OR lN6iiTU77ON EMIL R. UNANUQ M.D., Me/Nncfrods Professor of Immrwo~u Adop. Her- ..rd Medied Sclod. Wieto.. UNION CARBIDE CORPORATION. Nuck.r Dividoq Oak Ridge. Teaw. .,. STEPHEN F. VATNf?R, M D., A.wrlete lrofruor e(ediclnr, Harvard Medi- eal School. NMew englend Regional PrL wele Reseach Ceeter, Sowliboro, MeeF ocbo.etle; Mioriete /w Afed4nv, Peter 11eM 1/r1ssMer //oepltel. faoetow. IRENE Y. WANG. f w.D., A.NneM lro- /e.sor of Sesk and C/lwkef /mmwrol- op and Alkroblologr. Medical Uw1- verelty of Sout. Carolina. Cloer/e.low. ~ LfiR W. WATTENReRO, M.D.. rro/e.- aer of %rAef Uwivereily of MM- .esota Medinl~kook Mirrryoll.. GEORGE WEINBAUM. Hu.D., A1o.rl- enNu, rrfnwn..y Dlseo.e Section. AI- bert f_iw.leiw Medical Cenler, MYr de1pW. JAMES A. WILL. D.V.M. 1'w.D. Pro- /easor and CAeirw.r.. Drprtmenr of Yrterlnry Scknce, U.ivereily of WM- coweirt. Medi.o.. OEOROfl WOLF, D. Pna., Professor of rApiolotTkd CAtwd+try. Dererrn.ent of N.uriHon and Food Srknce, M.>W- eliueetb Institule of Tcc\nology, C.trt- (xidge, M.es. KOII YOSHINAOA, Pn D. Auoci.tr Professor of Anatomy. teio.etory of Hurnen RryrodecNon end Repod.c- tlrr /lofot7. H.rverd Medical Sc'ool. soeto.. FROJLCf Tlill I1re(o'Nbolm of norrwel e.d ectirNed .rworbqee Stod~ of 1~6 cf.r.aeAstlom of e.l.d inWetloe e.powr. L.ioee and dai- NKVY SyopkaeMd te.ob claospori.R tebdbe Nkaine-Ldro.d cef.g oao..ry veeodl- IMio. PAcae of cipreue m.ob and .iootloe on coro..ry e.d left .e.erk.dee dr•••dn OewNle dMerewoee In w/ra rltro wnaeo- eour liem of cRanicd trelt.oSe.. by Ii.wM IurMc.Uon et rmrtwo tanne of meure liver wlaos.el eylocRro... f-4130 InklWdon ef c.rclwoeennle by bea.yller Hioey.weN end rele/ed oorwprwde Lung .Milprotci.w Eelena urd the eRccM of cigarette rnole o./bY Iaeree(b. M re and IuweUonel eorreletlor tM Pl/D ulle ot /M kWy FJfed of vit.Wwin A on dycoprotelm @f'F lAesh is twrrnd and loranno.row eer pirelory eoiNlieliww Vitereiw A .nd silyooprouiw I. normal .nd preawoerow reWiretory eeNOellwn• wmerr sidecfoeyl trender• ..e, a ;or~ eerly a.rter eazyww for lledder eeecer FJ/ecu of sicotine ow prspe.ey 91 90 65100 7202

Completed Projects Following ia a ILt of the principd invertltaton, or inttituGooa, of p~oject. Ihat have been completed prior to the period covered in this lteport. Several of the individual. named are deceased. The titles and affilia- tbns listed Rre those in effect at the time the wort was completed. CLARENCE M. AORESS, M O., Au._ cl.a Ctinkd Professor of Medkinv. Universit of California Medical Ce.- terw t.ae znaeks. ANTIIONY A. AI.IMNP.SP PN D. Dl- rerror of i.boraro.les. The !lwke Re- \abililation Center. W1ite Plalow N.Y. ANTHONY P. AMAROSE. Psr.D.. /n- srrrcror ln Obstrtrks and Cynecolo;y, The Albany Medical Colkge of Unbn University. Albeny, N.Y. P. T. ANOFCAKOS. M D., Psr.D., Iro- Jessor of J Arslologr. Boston University School of Medicine. Boston. D. MURRAY ANOPVINP, M D. Unl- vetsilf of Wisconsin School of Medi- cine, Madison. DOMINOO M. AV1ArX). M D.. Irofrs- sor of Ihrn..roloer. Ilniversily of Rnnsflv.nia School of Medicine. Phil adelphia. STPPIIP.N M. AYRPS, M D, Dlrrrror, Crdinpulnron.ry laboratory. S.in1 VinceM's Hoyital, New York. OSCAR 1. RALCHUM, PsM O., Hasrlnlr Professor of Medicinr. University of Southern California School of Medl- cine, I.os Angeks. FRF.DERIK R. RANO. M.D.. Professor and CAwhnsen. Department ol Iartto- lioloty. The lotsne Hopkins University School of Hygkne and Public HeaUt< SaNieore. A. CI.IFFORD DARUPR. M D, RoAerl Nrnry I(il/kr Professor of lAlriol- orV, Harvard Medical School. Boston. tiRODA A. <iARNPS. M D, Pn D., rro- /rlor (AOlliou) of IArrinloRr. Cdo- r.do State I/nivershy, Fort Collins. FRl f)I RI('K W. !lARNF.I. 1.., M D, An.wurr Irn/rnnr of MrJi.lnr, llte Ii.hns Hi•p~ins Unwersity Sch.wl of Med~cint, Nalf.mote T. C. BARNES. D.Sc. Research Sckw- NW NYadelp\ia Stale HocpiuJ. P6p.- CA4Rl.*bW0. RECKER, M.D., Assoclue Professor of Pathology. Cornell I/ei- venhr Medkd Co11eSe, New York. R. PRE08RICK RLCKHR, PN D., Asso. ciare Professor of AnWonsy and Dkre- Wr. Laboratory of Irrlnaral Sclrncr, Duke University Medical Center. Dur- R.rq, N. C. RALPH S. BECKER. PMD.. I:o%uor e[ CRenrlsrry. University of NouMo~ Hauason. BENJAMIN !lELL, M.D., Dkecror Ens- erlr.u, Norn+otire Aging Srudy, Veter- ~ns Administration Outpatient Clink, (1o.tan. SAMUEL !!PI LET. M.D_ Director. DI- ririon of Cardiolorr, Philadelphia Ueneral Hoepiul, Phdadelphia. !•ARt11 RPNA('PRRAP, M D, Felyan hor r»or and ('Aairman. Drporrrornl of far otf, Il.rvard Medical School. RC»1on. IOHN A. BEVAN. M.D., Professor of IAorns.crolo University of Califor- si. Sctsool Medicine. Loe Anedes. BUDHDEV ls11AOAT, PM.D.. Professor o/ IA7dolftV. Saint i.oab University School of Medieine. St. l.ouis. CESARE !lIANCIFIORI, M D, DirlLlon of Cancer Rrsrrril, UniversN> of Peruaia, Pervltia, Italy. IIYI.AN A. RICKP.RMAN, M D, Ardsr- anr Professor of Afrdlrlnr, and AL- VAN I.. <iARACI/, M D., Conrrlt.nr In Medl.lnr, Columbia llniversity Co1• kae of Phnkians i Su.oeons; Odd- .ater Memorial I/ospilal. New York. R1()-RFSP.ARCH CONSULTANTS. INC.. Cambridge, Mass. !!1(1 RFSP.ARCH INS717U7P, INC.. ('arnhridge, Mau. 92 1 i FRED O. t1OCK, IM D.. Auod.re Can- cer Research ScirnNsr, llologk.l Sro- Non, Roswell Park Memorial IirtMMe, Springvilk, N. Y. OUENTHER RODPN, M D., Auocl.u Professor of AlydJcine; Aulnanr Dkec- to., General Clinkal Research Center, Temple University Hedtb Scienoa Ce.- 1er, PbiladelpAia. HERMAN V. BOENIO. PN D., Heod, Depranrnr of CAenrlsrry and DlocAene- bery. Spindktop Research Center. Lea- fnpon, Ky. IAMPS P. OONNeR, Pw.D. Professor liolop, Califord. InMNwe of «hnoloar. Paaads... WALTER M. ROOKER, hr.D., Iro/es- soe and Nrad. Department o/ IArin.- rolos7. Howard Ualvenill. Waef.iy- sow, D.C. TOM O. 11OWERY, Pr.D., Research Iro/ersor. Pesticide Resldw tatror.- tory. Nortb Carolina Stat. Colkte, R.kiat.. OEOFFRlY L. BRINKMAN. M.D., Ar .ociore Professor of Medk'ine, W.ynr Slate Usdvereity Sebool of Mediciws, Detroit. ROBERT E. BROOKS. Pw.D. Asrocirre rro/e»or of Pathology. University ot Oregon Medical Scbol, Porlland. BARBARA R. BROWN. h1.D.. CA/rf, Eaperlnsenral IsPcAWry, Vderw Ad- miniMrNion Hospital, Sq.IvedK CA. RAYMOND R. tIROWN, Pw.D, lro/es- sor of Cllnkal Oncolop, University of Wisconsin Medical Scboo/, MadBoa. JOSEP •ROZP.K, Pw.D., ho/rner .nd Chairman. Department 01 Psychology. Let.isle Universily. edYe!•ens. Pa. SUE sUCKINOHAM, M.D. Assistant Iro/r»or of Idiabkr. Columbia Uni- .er.itr Collep of Pt.rskiw a Sur- seona, New Yak. <iP.N1AMIN st1RROWS, M 1)., As«r- dose Iro(esuar of Medklnr. Unhasiy of Chicaao, Chk.ao. E. M. tlUiT. M D., Chief IrrAoloO.r, las Anseks Coualy Oener.l IlospnaL 1 os Angeles. RICHARD Il t1YPRRt1M, PwD, ho- of CAembrrl, Micsip. Stae niverutr. P.u landns. C SISTER M. P.MILY CAH/l.l., PuD. CA.kn+o". De'ortn.enr of CArns4rry, Regis Colk~e, Weatoa, Mw. BRUCE P. CAMERON. M.D_ lhsD, Howard Hu~bea Institute. Univenhr of Miami Sc6oo1 of McdkM.. Ml..` Fla. WILUAM H. CARNES. MD., Udat- sily of Utate Cdkge of Medicin., Sak Lak. City. MARCUS N. CARROU., la_ P..D, Chief. D/risiew of lAsnsocoloPy, TY Rraokdale Ho.piwd CeMer, RrooklM N. Y. WILLIAM ALVIN CARTRR MD AsdAant Professor o/ Afrllclwr .nj Alk+oll(olotP, The /otssoa Hopkins U.L ver.My Settool of Medicine, .Riwmr+. LEOPOLD R. CERL'CEIb, PMD., lrr ~iuor of floc/irnrislry and Nwr/Moq, University of Puerto Rico ScAod of Medie3na, Sao Iwr. CHILDREN'S HOtLtNTAL OF LOd AN- OP_t.PS, Lo. AnReka SANFORD CIIODO6H, MD.. Assistant Professor of Mediclnr, Tufu Ualw- si/f Sctsool of Medicine. lsoeto.. NAITBR M. CHOPRA, Pr.D. M/rr "r e/ CArwsLr.y, Nortt. Carolina As- rieulard and Technical Suta U.leea eitr, are.wsboeo. WIt.LIAM O. CI.ARK, Pr.D_ D1.ecr.r. rqcllol6.raw.rolo~~ Research l..iort Iory, Veleran Aderi.iMraMlon Ho.pltal Sepulveda. C.1. HANS T. CLARKE, DSc. rroJrur of IiocArnrisbr, Columbia University College of Myekian. & SurReony New York. JAY D. COPFMAN, M.D. SrcMon Nrod, Peripheral Vascadar Delarrnrenl, Uni.erswr I/oylid. Iloatow. • DANIEI. COH@N, D.V.M., M.P.H. A.r sraaM Professor o Yeserlnr) Epi- denrblolP and Iuk~e Hea1rA. Uaiver- sitr of Penn,rlvania Sckrod of Vel.r- inuP Medicina, Ptsiladelp4ia. IULIUS H COMROP, 1., M D, Dkac- ror, Cardiorscsdar ResercA lnulrwr, IlniversilT of Cdifornia Medical (:eo ter, San Franeiaco. 93

DEAN M. CONNORS. MD. Assoclre Dutcror, Dep.rrwttwt of Laboratory MrdKlwr, SI. M.ry i Hoepual, Madi.oq Wi.. PlflLll COOI eR, M D. Clinical rro- /euor oJ Svlery ewd Dlrector. Srjf- cd t.lorwtory of Crlluli rAy.iolotly Albcrt Eiostel. CoUep of Mediciua of< Yc.lw.e Udverd('• CA/e/, Srrgk~./ Service. Veterar Adminie(ratb. Hao. pilal, Ttr Bro.a, N. Y. JOHN 0. CRAIOHP.AD, M.D_ rn/er t~ C~Aolot~ Univereily of Vet- p of edki.e. B.rUyto.. RORHRT L. CRAIN, Pn D, Au/.ewt rro/rssor of Soriolop. UnhereMy of Clikago. Chicetlo. T. TIMOTHY CROCKER, M.D.. rroler- eor o/ ALdkine. Universitr of Cdl- fornia College of Medicine. Irvine. CARROLI. P. CROSS. M D.. Auoc/wre rroJrssor of Alydlriwr .wd Nwwww rAyslolop; DYrctor. Stctlon o/ rut- swwery E(rdiciwe. Univcrdq o~ CaN- fornie School of Medicine. Davie. CRCIL E. CROS3, RrtrercA Deo.rrment. St. lo.eph IlorpW. Burbaat, Cal. ALBERT DAMON. M.D., Pu D., Lre- trrr ow AwtAroroloty; RruncA Mso- clo(e /w Alydk.l AntAropolor', Pea- body Museum. Harvard UniversBy. Cambridge. Mar. THOMAS R. DAWBER. M.D., Auociue rro/rssor of Mrdkiwr, tiuMos Uaiver- .ity School of Medkine. Boelow. R. F. DAWSON, rw.D., rro/essor of 1a- owy. Col.mbie Uaivenity. New Yort. JOHN P. DFLANEY, M.D. Nw.D, Ar- rocine rro/rssor of Sr ery, Univenity of Minnesote, Minneapo~ie. ANDREW 3. DIBNP.R, Pn D., Eeea rtire, rrycAo-RrurcA, T1e Ap Ces- tee of New England. Ine. Bouon. EDWARD F. DOMINO. MD.. rro/es- sor of rAarwrorolop. Univereily of Michigan. Ann Arbor. RAI Pit I IN)R/ MAN. hrll, fN.rrrar nl I rA...m...u. W..rtr.rrr I...rnJ.u.rn I..r 1 .pr.m.rru.l Bn•1,•r. \hr..d•ury. N ... H. FRED DOWNEY h D., Auist.nr rro/e>aor of rAysl-dojy. Univer.i/y of Teau Heal(S Science Center u Dallae; Director, CYdlOV6lfYlr Rrse4cA, Car- diooulmoeary Inwitule, MetAodip Ho.- rital of Ddla.. D.Ru. JAMES 1. DYAR, rr D.. Arn4unr rro- esaor o Qiolop, Bellarmine College. le. Ky. RICHARD H. PARLF, M D., CA/r/, rufwwwary Frwcrlow Laboratory; As- dstowr Professor of A/edklne. Unlver- rly of Clak.tq Ct,k.Ro. JOHN W. ECKSTEIN M D, Aubtawt rro esxr oJ huerwe) Alydirlwr, Sute U y of lowa Cdkp of Mcdl- dwe. lowe Ciq. BPRTRAM EICHEL, D.D.S., Director, fnpltore of Stow+otolotk.i ReurcA, Scier.oe Re.oureee Founde(ion, Watcr- Ww.. Mer. HYMAN ENOP_I.BP.RO, M D. Anend- Inir f'Aysklow, Ccdare of Leba.on Hoe- pi(al. Los Angeles. CARLTON K. ERICKSON. M D, As- soclrr rro/esror of rArmxolotr.wd Torkototr, The University of Kaneae ScAool of Marmacy, Lawrence. HFNRY 1. PSBP.R, fn D., ResrorcA Iwr- nruwologlsr, Ma.on Research Ins1i(ute, Woruaer, Mau. 1O/1N R. PSfpRI-Y, M D., A»oclue Professor o/ rrAdory, Usiveraily of ~aRo t rM:ter School of Medicine, fto- HUGH 6. EVANS. M.D.. D4ertor, De- prswuwr of rcdietrks, lerLA llospi- Id end Medical Ccnler of Brootlyn, iiroot/yq N. Y. HANS 1. EYSP.NCK, <+w D., l7Sc., rro- Jeswr of rs)cAolotr. In.(i(u(e of hy- cAlalry. Unlversiqr of lor«lnn, l.on- dos. HANS l.. FAI.K, Pit D., Ad/wnct Assocl- ote Professor of rotAololy. Universily of Soutlcrn ('alifornia ScMtul of Medicine, Los Angeles. DANA l.. FARNSWORl ll. M I). l/rnry K Ohvw rrafrnor .r/ Hrc.rnr and 11„nr..r, f/nnv.utr Ilr,JrA Srrvn"r. 11.r.euJ I/m.a.ny. ('.rnb/rJ{e. M.u. v4 r r I i GAD FPJNSTEIN, MD_ Senior Ltc- rurrr /w IlocArwrlsnr, The GeorRe S. Wiee Center of Life Sdencsa. Tel Aviv Ueiveusily, Tel Aviv, ler.d. PRANK C. PEROU3ON, 1n, M.D~ CAelrwrew, Dtprnwewt of rArw.aco/- oly. Tle Alba.y Medical College of Union Uehereity, Albany. N.Y. ~ THEODORE N. PIINLEY, M.D, D/r.c ror, Pulmonary RerrrcA 1alorrtory, Mount Zion Hoqitd, Sew Prucieco. WILLIAM 1. PISHBPIN, M.D.. CA41 o Er11ew.ldop, CYe.p Board of / eal(R Chicago. eDWIN R. PISHER, M.D. Direcror of L.frorotorbs, Stadydde U/~it~ r trJOr of rNAOI J ~. ~a( Pittsburgh Sct~oot o( Ir buryla. RUSSPLI.S. FISHHR, M.D. Usi.er.B~ of Meryl.wd Seftoal ef Mediei.e, /iewre. B. L. FREEDLANDER. M.D D/recwr of Cancer RrurcA, Morwl I" Hof plel and Mcdkd Ce.ler, Sew Pre.- cieco. FREDERIC A. FRENCH, A.R. Dirse- tor of Cwwcer CArwruAeryy R.sron~, Mount 73o. Hospital .d Medkal Centec, 3am Frand.co. JACK FREUND, M.D. AaWowr rro- (tssor of rAorwr.colop, Medical Cd- kile of Vlriti.le, RieJr.o.d. OII.BERT H. PRIEDLLL, M.D CAk, of rwtAotog), St. VleoeM 1iYor~U.~ Worus(er, Mer, H. HU011 FUDflNBPRO, M.D.. rro%r sor of M.dlciwr Univsr.ity of Call- fornia Medical C(eMer S.si Fre.eMcol Professor of socter and Iwuwr- nolop. Unf.erilty of (fanle. Rert.- ky. ARTIIIIR Fl/RST, M.D., DYrrtor, Ar- stltra of Chemical l/olody, University of Sa. Francisco. San Freacitoo. MURRAY R. GARDNER. M.D.. Auo- c/ort rro/essor of raAo%r. Uaiver- eky of Sout`er. Califor.ia School af Medkine, Lo. Angeles. OF.OROP O. (1PY, M D., Director. F/w- wrr llorsll Cancer Rru.rcA ImIoro- tory; Asr.w-/ar rrofn.or of Sur4rr~~ lte loAne Iloytin. l)nivereity Sclool of Medicine, Bdtimore. THOMAS M. OOCKF_ M D, Araec/rr rro/rssor Of rFerewllvt M.dk/we w1 Cowurrw/ty Health. Scion Hall Col- M of Medicinc ewd Dcairry. /ar.qr , N./. DAVID M. OOI.DPNBERO, Sc.D. M.D, Associate Professor of %tAol- op. Temple U.ivereily HedtR Sd- eaw Center. rW.delRNe. PAUL OOLDHARHR, D D S., Atacbe rro/.uor of rniodowtolotf. Har..rd School o[ DeMal Medicine. Boe(o.. LEONIDE OOl DSTEIN, D3c., As.r ci.q rro/tssoe o~/qcA1.try, IwMi1nM fo. Mew1d Ik.ltlr Sciewocv. College e1 Medicine & Dentistry of New /aee1, RMgere Medicd School, taiecuawer. IRA OOR1y M.D, rrole»o+ of ratAol. ~~ Reno. UnhrenNy Sctrool of Me04 eMr• CAfe/ of Lior.tarr Serdce. Veurnr Ad.nWMratio. lloyital, Wu1 Roalury, Ma.. OERTRUDt' Y. OOTTSCHALL, IR.D. Arsluowr Professor of I/ocAr/wWry. Collrwbi. U.iverd(y College a[ Myei- derr L SurReorlr. New Yort. A. CLARK ORIFFIN, Pw.D, Neef, Dtprtwwwt of AlocAewd.rry. M. D. AndenoR Hoapital and Tts.wr Lrth Wte, University of Teaae M.dlcd Cewr, Houelo.. ARTHUR L. GROSS. M.S, Sewlar 9/. cAewdst, SowAweel Research IrNuY, Swn AMo.io, Tea. MORTON 1. GROSSMAN. M.D. rr.D. Associate Cllnkll rro'fYor O we~- clwr, Udvereity of C.lirorwla Cenler, Loe AnRela. CARL C. ORUH7lT, M.D. Pw.D. A. •oerere /w Physiology and f'A.rwaeof- odr. U.i.ereitr of Pennsylvania Orea- rde 3ctrool o! Medkiwe. MiladelpW. /06EPH 1. Ol/ARNP.R1, hr.D. Anewl- Iw# A(kroblolog/L; D4ector, Afkvo(rI- obp LoDoraortrs. l on. Island lew- itA-Ililleide Medical Cenler. Queta. Ifo+pita) Ce.1er Afflllatiow, lawraica, NY. FRANK P. (1UTIIRIE. M D, rrofo soe of EwtwwoloR y, Nortb Carolina State Cdktle. RakiRlr FI. B. HAA(1. M D.. rrofrsror o rA.r wr.colofy, Medical Colkoic of kfilaia. Richmond. 95

I i P. 1. HADDY, M D, PN.D. rrolessr and C'h.bm.n, De/rtmrnt of rhysl- dotl. University of Oklattana Medical CeNCr, Oklabom. City. JOSEPH H. HAFKENSCHIEL, M.D, Direao., CrdioPa>lmonry Unlt, Tte Lankeaau HosPital• Associate in Medl- crne, l)nirersity of ~ennsylrenia School of Medreine, PhHedclPbia, BERNARD HANP~ PwD., Prolrsaor of Ne.lrh Science. C ifornia State Uaver- sity, Nortkvidge. RICHARD 1. HAVlI., MD Asdst.wt Professor of A(ediclwe. UnZversit of Calrfornia Medical Center, San Frrat cuco. HERRF.RT R. HAWTHORNP, MD, (-h.rrm.n, Drprtment of Satfery. Unirersiry of Penw}~lvania Or.duae Sckoo/ of Mediciee, Kiladelobia. JOIIN A. 1/AYFS, M.D, .Nsociur r.thorosiu, Mallory Institule of Pe- tbolooly, floatoe City Hospt.l, Sosto.. CLARK W. HEATH. M D., Professor of Mdinne ond Di,ertor of Health Srrr- krs, Tufte Universiry, Medford, Mass. PAULINE Hf?III?R, PB D, Research Associate /n Cytology and Cyrochem/s- ny, San Francisco Institute of Medical Sciences. San Fra.ciseo. LAWRENCE L. HESTER. 1.., MD, lro/e%wr ond Ch.irm.>,- Department o/ O6nnrlrs and Gynecology, Medical C.o1kRe of South Carolina. Cbarkuo.. EBBE CURTIS HOFF, MD., PND., Pro/rur and Chelrn.en, Diriylon of Psychiatric ResereA, Medical Colkp of Virli.ia, Rkkeao.d. RUSSELL L. HOLMAN, M.D., Loald- .ne SIa1e University Sckrool of Medi- crne, New Orleans. OLES A. 11OLTL'RMANN M.D., Re- urch Scientist. Lo1rnJ Liloraory, l)nivenily u/ Notre Dame. Noue Dame. Ind. FRNDDY H(1MbUROFR, M.D., rresi. dens rnJ Drrecror, Sw Researeb (ns11- tu1e, Inc., Cambridge, Meu. k/1NFR1 W IIUI1, Prr1), Pr.drrror of Sud,.rr.ul Sruenrrr, Flonda Stalt (tnrvrruty, I.tl.buxe IIT RESEARCH INSTfiUTEE, C.'6icap. GEORGE JACOBSON. MD. rroleswr ead Nr.d. Dr/r tmewt of Rodlolosr. Uelverdl of Soutk.er. Califorda Sclod ol Medidae, Loe Aegeles. JERRY HART JACOSSON, M.D. Dl- recrr, Division of ElecnoPh"iofoty, New York Eye .nd Pa. l.firmary, New York. JULIUS H. JACOBSON 11, M.D, Auo- clre rrolrsso. o/ Sw`rry .nd Director J Srj/c.l RrwrcA, University of rrnoat Cotlcp of Medicine, flur- Nn~tow . PETER If. KNAPP. M.D. ResrrcA ProJruor of Psychiatry. doeloo Uwl- versity School of Medkiwe, flosto.. KENNP.TH P. KNUDTSON. M.D U.i- versity of WaebioRton School of l/edi- cix, Seattle. ALVIN 1. KOSAK, PND., Associate rro/e»or of Chemistry. New York Unirersily, New York. RO<iFRT A. KUHN. M.D.. Aa.ocl.tr Pro/essor, D/rb/on of Nrrrosrrtery. New Jersey State Cofkp of Medieine, Jersey City. AVERILL A. LIEBOW, M D, CJni• www, Deprtn.rwr of Pathology. Y.b Uniwsil~ 3chod of MedkMe, New H.vee, Cor. ESTEN O. LINDSETH, MD. PrD, St. JosepR"t Hwptd ReasarcR Laboratory. St. I aul, Min.. ROBERT H. LINNPJI., Pr.D AsncL ote Professor o/ CArwds(ry, 0nivertlty of Verooal, <rurlieRlow. HERBERT L. LOMBARD. M.D., M.P.H, A/Y/ne, Cr+cer Research /n- sfifWe, New England Deaconea flo.. pitsl, liodo.. MARVIN KUSCfINP R MD New . , ., Y k U I. P. t.ONO Pr D Professor of PA.n MURRAY E. JARVIK, h(D, Assocl.re ho/ia.or o/ PArwucolofl. Albert 1'ia- eteln Cdktc of Medkir of Yeshiva or niver.ity Medkd Center, New York. CHARLFS W hRELLE Pr D AuW- . , ., wsocolofty State University of lowe Colk{e Q Medici.e, Iowa City. Uwirersity, T)e tironz, N. Y. . , . .. .ni Professor of Ew.ironmenrof Ny- CLAYTON O. LOOSLI, Pw D., M.D. tienr leffeteo. Medical Co1k e Prila- NostiwRe rro/enor of dhdicint .ad OSWALD R. JONP$, MD. St. Luke i , R , delpbia rothology, University of Sowlser. C./I- Hoepitd. New York. . fornla School of Medicine Los Angeles I , . ANDREW A. KANDUTSCH PN.D AARON J. LADMAN. Pu.D.. Professor DONALO R LOURIA Assecfue M D . , Stee Sclenrlu Tb Jackso. lAbota- and Ch.irmen, De/rtment of Anr- . , . ., Pro%scar of Medeclne rnefl Uaiver- C , tory, flu Harbo., Me. omy, The University of New Meako School of Medkio., A1bur,ueraue. , o .i1y Medic.l Cdkge, New York. ARNOI D R. KAPLAN. PrM D., Dlrec- THOMAS C. LAIPPLY M.D. Pro/es- KENNETH MERRILL LYNCH. M.D to., 1.oAor.rory of Mrdlc.l Grnrtres, , , sor of Potholotr NorUweMere l)d- Sc.D. k.L.D., rrolrtror Emer4re o~ Cleveland Psychiatric Insriquwe and . versily Medical Sctiool Ckok.Ro. Pathology Md Chenceffor, edictlM flu.ppul, Cleveland. , Colkp of South Cardi.., Cir1..1w. ATTALLAH KAPPAS. M.D. Professor and Senior /•hysk4n, The Rockefeller University. New York. , HRATCH KASPARIAN, M D., Anlx- ew/ DMedr, Crdiorascdar L.Aore- (ory; iwsanctr in Medlciwe, llaboe- twenn Medical CdkRe .ad Hospital. Philadelphia. ELIHU KATZ, PN.D., Associate Pro/rr, sor of Soclofoly. University of Cbi- caffo, Chicago. Sf11Rl.EY L. KAUFFMAN. M D., ho- lessr o~ lathdory, Slale University of New ork 1)own.rale Medical Cce• ter, Rrooklyo. ANCEL KEYS. hr.D., Director. Jso.o- tory of Physlofoslcd Hygiene, Univer- sity of Minnesota School of Public Hedtb. Minneepulis. /l)Sf?P/I i/ KIkSNl;k, M D. rr•.lets•.r ,n/ Medicine. l)nirersily of Cbicago Scbuul of Medicint. Chicago. 96 ( MICHAEL E. LAMM. M.D. Professor of J'rhdoj*y, New York University Medi- cal Cerwer, New York. PAUL S. LARSON Pw.D_ Hees Pro/es- sr of rhrm.coWq, Medical College of Virginia, Rickrnond. ' ROGER K. I.ARSON, M.D. Chief of Mrdklne, Fresrw CowMy Hoyk.1, Fresno, Cal. OUSTAVE A. LAURENZI. M.D.. Chief of Afediclne, St. Vincent F1oyiW, Worcester. Mw. EDWARD I FETF. Pu D, DSe lroler- sw of ('hemisrry. University oI Mrnne- • soda, Minneapolis. RIC/IARI) A. I_1?RNI!R, M D.. Associate MemAer, Scripps Clinic and Research Foundation- 1.e lolla, Ca1. CIK'11.E LP.l1( IfINNf!@R(3PR, PND, • f/eod, DePrrmrnt of Crtochemisrry, Swiss Inartute for Eapcermcntal Can- cer Researcb, Lausenne, Switrerlasd. INES MANDL, PN.D., Assistant Lo/is- nor of IfocAenslary, Cdumbia Univa- eity (.'ollep of PhydeLw. A Surgeons. New Yotk. JOHN H. MANI/OLD, I.., D.k/D rro/tswr owd Director. Department o) E.rAdory and Oral DieRnosfs, New Jersey College of Medieine and De.- lis(ry, Jeray City. DAVID E. MANN, PN D, Auoclrr Pro%swr of rhrmocolop, T.wr~le University School of PYws.clr PfO., dclpria. /()11N P. MAN()S, M D, lnsr.rrtor le V1rdogy and s.crnlologr, Medleal Colkile of South C.rolina, Clarkaton. CIIRISTOPfIFR M. MARTIN, M D Auut.nt Prolrsro.- of Medtrlne enj Drrrctor. Dirtsion of Inlerrioas Dis- e.su, Sc1ow 11d1 Colkp of Medicine. Jereey Cay. MASON RFSEARC'll INSfTIIfiE, Worcester, Mass. 91

~ . DONALD 1. MASSARO. M.D.. Aswci- ae lro/euor o( MeJicine. Oeorge W.>Din/ton Usuvenll f School cf Medicine. Wast,inpoa. D. C. CHARLFS McARTHUR. rw D., t.y- chologiu. Unlrersky Health Serrkes, llu.aed Usivcreity. CamMidge, Mua. CHARL.ES LI. McCANTS. PN.D. Aactr- clae Professor o/ Soib. Nort\ Carr lin. Stale College School of Agriai. Iwe. Raleigh. HENRY C. McU1LL, la, M.D, Acq~ H.af. Drp.rrmenr of haAolop. 1 at~. .iew. S1Ne ISnive..Mf School o/ Med4 ciwe. New O.Ie..e. /IENRY D. McINTOSII. M.D.. lro%r sor of Mrdklne owf Director Crf/s rascY/r L1ar.rorS. DrAe Ijwiverclty Medical Centu, DwAa.n. N. C. FnRDF. A. McIVER. M D.. Asaor/w Professor of lrhology. Medical Cd- kp of Sonth Carolina, Cloarleelow. EDWARD McKEE. M.D. Professor anl Acting CAdrnwn. Drp.rlmem Pathology. Medical College of Sout~ Carolina. CharkMon. KELLY T. McKeE. M D.. Auocfate Professor o~ Aledkiwr. Medical Col- Iqe of Sout Carollna. Cbrkeuow. VICTOR A. MeKUSICK. M.D. rro/es- sor of AfelicJne. Tioe lo\ne 11optlne University School of Medicine. Sdtl- nare. ROSS 1.. McLEAN, M.D.. AuocWe Professor of Afrdclne. Fmar Uaiver- .ily School of Medkine. AU.m.. WILLIAM P. MtNARY, la., PMD. Ar sorWe ho/es.or of AnaorwT. LloMow Uni.ercMy Sct,od of MedidwR, Rotlo.. NEAL L. McNIVEN. hMD_ "e Wor- oerter Porwdalott for EaperioeMal Biology. SIKe..bmwr. Mass. lULIA MEYER. M.D., A.aociiru ho- nsore/ Or+d Pathology. UdversNy af Minoie College of De.tkuy. C'lieap. DOV MICHAELI. Pw.D, Assisrant rro- tsor of liocAemlury on/ Sw~rry nivereiq d Cdlfornia ScAooi o~ Me.hcine. Sea Francisco. eFRNARD 1. M111 P.R. M D. Asu.tem rrofrrsw of Awaeom., )e//nwn Medi- ul (-olle/e. PfdadelpMa JAMES O. MILLER, M.D., ht.D., rro- /euor of rrScAi.tr' and rsycholo 1; DYedor Mental HcolrA Research Sn- nlnui, (ldven7ly of MkAiAan. Ane Areor. CHARLES MITTMAN. M.D, D.recror, Department of Respiratory D/xeses. p1) of Hope National Medkal.Ce.- br. D.uw. Cal. HUGH MONTOOMERY, M.D.. Asso- clere Professor of Medicine. University od lhnr va.la School of Medici.e, /fMdclia. P. O'R. MONTGOMERY ). , M D Iro/iuer o/ lrAelot~, ~hdrer.Nr o~ Tcer Sowlwaen Medkal Sc6oai. DeRas. OPAROE !. MOORE, MD, F+r.D., n/- rccror RoeweM Park MemorW /rb t.w. ilaiab. N. Y. KENNETH M. MOSER. M.D., Assistant Professor of Me/iclnr. Oeocge/ow. Uai.eniY~ L~Iedk.l School. Washiy- to.. D. C. HURLEY LEE MOTLEY. M.D.. lro/ea` so. o Afr/kine and Director. Crli.- Res~rnor ~rnor~ Lboraory. Univercitr of SouMerw California School of Mcdl- cine. Loe Angeles. EDMOND ANTHONY MURPILY, M D., Sc.D. AuocWe Professor of Iiosurbrkr and Medicine. The /obee Hopkins University School of Medl- eine. llaitisaore. W1LLIAM S. MURRAY, Se.O.. Senbr Sre/ ScirwtW. The laAtwn L..twr.- lory. Rar H.rbor, Me. RICHARD L. NAEYR M.D, Professor onI CArbww. Dep.t rmenr of r.rAol. ' op. Ttwylv.sti. Stat. University Col- kge of MedkMie, Her.ttey. AL.tPRT H. NIDEN, M.D.. Iro/euore/ Mefklne, Drew roetgrad.a/e Medical School and Itehertity of Sowdlcr. Celiforni.• Chief. trbnow.ry Disease Section. Manie Lwtter KLA Hospital. Lo Andek.. OAK RIDOE NATIONAL LALIORA- TORY, Oak Ridge. Te.w. L)nNAI D M. IACP.. Iht D. Irofnsor of lAysiolop .nl D.recrw. Jnsurra Z Cel1rl.r RrtrrcA. Univelsitr of lw.sla. Linculn. 98 i` I i t ALBERT S. PALMER. M D, AscW.n/ TIMOTHY 1. REGAN. M.D., h%s.ar- ho/ruor of Pathology. U.ivereily of of Me/klnr; Director. D/rWow o/ Toledoq Toledo. O. Crdiowcr/r Diuoxr. C o~olk~. Medkiae and Dcwielry of New lerxf. ROSE MARIE PANOSORN. MS, Aa- New Jersey Medical Sc6oo(. Newark. .iatenr Food Tec w and [.ensrer. Depa.rmrnr of Food Science ond TecA. WILLIAM REOELSON. M.D. ho%amr naotr. Uwercity of Califor.iK Davis. and CA.Mim.w. Drrarrn.enr of U.Ik.l Owcoloty. Mediul College of Viql.l.. JOHN W. PARKER. M.O. Aarocl.fi RieMwnd, Professor of rMAoloty. UnivenK>• of Sowrer. California Scbol of Medl- H6RART A. REIMANN. M.D. h./e.. ciwc. Lae Angelee. sor of Medicine. Hab.cnnat.. Medical MARY STEARNS 1'ARSHLeY, tiD., College mil Hoopkv. Philadellw- Assiu.w/ Professor of AwNOiw~ M Ob- ROLLAND C. RP.YNOI.DS. M.D_ A.r «errks and O,neco%p ColwwUl. sbaiw rro%uor of r«Aofogs. Uwivu- UnivereMy College of Myejcl.we R Sur- cN~ o~f Teaae So..tKweereen MedkJ powti New Yo.' ScAoW, D.R... EDWARD W. PELIKAN. M.D.. CAi- VICTOR RICHARDS. M.D.. Chief of wuw. Dqanmem of rAriwacofoSl SrterS. Meebyteria. Mcdkal Ce.w, and E+perlwanral TAerornrki. LloMOw Sao Francisco. UeivereitS School ef MediciaS Roelo.. ARNIS RICHTERS. r.D, Aseocire MALCOLM C. PIKE. hr.D„ Professor Professor of l.rholop. Usal~~~ e/ Communlry /feliciwr .wI redlN- SowtwwC.Wa.i Seltooll rki, Uwirer.il~ of Southern CJUa.i. La. Angeles. School of Medicitse. L.oe Angeles. WILLIS H. RIESEN. Prr.D.. Sewl.r 11e.. OTAKAR 1. POLLAK. M.D.. hr D, eAcwdst. LI/r Sclewcer Division. IIT liiecwlre Director. Dorer Medical Ra Rese.rcl IaslKwe. Chicago. KYch Cener. Inc. Do.er, Del. DANIF.L S. RIFKIN. PN.D, Asulw.nl MORRIS POLLARD. l1M.D, Director Professor of Chemical /b/oty. Tio Lortrnd L.1or.rory. UeiraeKS oj Rodefeqer University. New York. Naare Dame. Nare De.ne, Itd R. H. RIODON. M.D, Professor of %- C. M. rOMERAT. h.D.. Director of rAoloR) , University of Teane tNedkal Iiologk.l ResercA, Pasadena Fowd.- R+nwc~, Qal.erew. tiow (a Medical Researdti P.e.dem Cal. SYDNEY C. RFI-LNRERO. AtD, hofeiaor of locterlotop, Utri.eesAS S. N. PRADHAN, M.D fw.D. ho/n- of SowRerw Cdi/arw4. La Angeles. sor o/ lArm.cdoS~, ~low.rd Uwiver- wy Co1kAe of Mc~ci.e, WrRinpoR BENSON e. ROIy M.D.. Assoclate Iho- D.C. /esso+ ol Srsery; Chief, Cardiac San ~rry~ U.iverdl, e( California School H. R. rRATT-TIIOMAS. M.D, rro/er of Medkine. S.n Francisco. sor o/ Pathology and De.n, Medical College of South Cardin.. CA.rlnton. IOSF.r1/ H. ROOERS. M D, Holy Nawe of lecae Hoyild, Oodedso, AL.. PR(K'1?Ct A INTTRIIMENT CORfA- RA7fON. droollyn, N. Y. ROBERT C. ROSAN, M D.. Associate Professor o Pathology and re/l.nke. MARTIN S. PROTT.1?L., DD.S., Chief. SI. l.ords sdvercNS Scttod of Medb Drportmenr of Or.f Pathology. Newark eine' Associate lsAoloSir, Cardioal City Hospital. Newark. N. l. Oktiwow Ma.wW HaFNal for Ch0- dre., St. L.o.ia. WAI.TF.R RFDIS('H. M Dm As.ocWe Lolrsvn of Clinicrl Medicine. New CIIARI FS L. ROSN. hr-D~ Clink Dlr.c- Ytwt Univn.ity ScUorrl of Medicine. Ior; DireNOr. Normurlre Aging Study. and NYU Ke.eaccA Service. Ooldwaer Velerane AdminlMrs/k+w Owpatkal Menwrial Ilospiul, New York. Clieic. Bowoe. 99

0 JOHN R. ROWLANDS. Pe.D, Sraff SckwNst, Soutbweet Research Innitute, S.. Antoeio. Tea. SF.NIAMIN A. RUSIN, Pw D., Assbrant Profnsor of f•rslk Htalth, Be~lot U.i.crsity College of Mcdicine, tlou- ston. RONALD P. RURIN, M D. Professor J thrmecoloty. Medical College of keeinie, Richmond. HENRY 1. RI/SSEK, M.D lrrsidrwt, T1e Ruseck Foundatioq . ~nc, Stelera W.nd- N. Y. W. C. RUSSEII, M.D_ Univershy o[ Te.es Medical Center. Houston. WAYNE L. RYAN Pw D., tarolessor of Ilochem/srry. U4versitP of Nebraska College of Medicine, Omebe. PETER F. SALISRURY, M.D. Pw D, Ilyd, Iwtrwsirr Treate.rnt Center. Seint laeep6 Ho.pi(el, Burbank. Cal. PAUL SALTMAN, h( D~ Assistant rro- Gs.or of Siochnwist.~ and NrtrUiow, avverskr of SoWhert C.li/ord. School d Medicis+e, l.oe Anitcles. ULRICH H. SCHAF.PPI. M.D-, Diree- tr of NewoOhrnracoloq, Mason Re- se.rcA Inwitute, Wroester, Mess. IOROFN U. SCIIl.EOEL, M D., Pn D., lrofessr and Chokwtaw. Drrartmrnr of Srr y, Tul..e Udversiq School of Medicine. New Ork.ns. ALVIN R. SCHMIDT. Pw.D., Director of Cornwling, Thtls University. Med- /or4 Mass. ISAAC SCHOUR. D DS., PwD., DSe. Dean. University of Illinoie College ol Deairry. (.'hicyo. SCRIPPS CLINIC AND RESEARCH FOUNDA710N, l.. lolltt, Cal. MAURK'E S. SFOAL, M D., CUakaf Irole»or of Medicine. Tulls Univer- shy School of Medicine; Director. Dr- rerrmrwt of Inhalation Therapy. soF lon CNy Ilospiul. S.ntun. 111('1/) tl!VPR1. W 1). DorrroW and hr.w lwuurts ./ A.rn..wr awd Ia0Aal V` r. I)/tnMM of (.Mer Reur.t(~, llm.u.1y of Pervt.., Prru.u. It.iy CHARLES ft. SHAW. MD. Chief. Src- riow of Medical Grwrrics, M. D. Aa- daww Hospit.i .nd Turnor Inetitule; Professor of Iiololf. The Ur.iversity of teas,e at Houslon, Houston. CHARL9S E. SHERWOOD. M-D. As- sia.or Profr»r of Rodiolo,r. U.iver- sily of Rochester School of Medieine ..d Dentistry. Rockster, N. Y. SHOJI SHIDATA, M.D. Pw.D. hofes- •or r~a*•cololf. University of HawA School of Mediei.e, Honolulu. DAVID L. SIMON, M.D fwstrrctor in Internal Mrdklrv, Ci.cZ.a.U Oeserel Hosritdk Cincin.etl. ERIK SKINHWI, M.D., Chief. Drrart- srsewt of Neurology. Riqeb/ert liospi- tet CoPe.Y.ae.. THEODORE A. SLOTKIN PN D. Ar Nstanr trrofessoe o/ lLarrnacolop, Duke University Medical Center. Dur- h.ns, N.C. GEORGE W. SME'ITERS. M.D. Asso- cLre M trholoTr, Northwestern Uni- versily Medical Scbnl, Chicago. GENE M. SMIT Pu.D., Assbtant rro- /ruor of rs)c otf. Harvard Medical Scbool, M.sucbusetts General Hospi- teL Routon. LUCILE SMITH. PN.D. Professor of Iiochrmistrf, Dartmouth Medic.l School. Ilenover, N. H. SHELDON C. SOMMERS, M.D., Dirrc- ror of tilorrries, Lenoa Ilill Hos- Pita, Cl/sdcel Professor of rrholoq. Columbia U.ivereilY College ot Physi- ei.ns A Suryeo.e, New York. ERNEST SONDHEIMER. PM.D., Asso- elrt Profeasr of Ilochrmisrry. Col- kRa of Forestry, Stus Univerrty of New Yak, Syracuse. T. M. SONNESORN, PM D.. Distiw- .d Service lrolrssor of Zoolol~, St.nlsA.ddi.n. Utsivereily, blowninS(on. SAM SOROF, Pu.D., Head. Dyartmrnt o/ Macroatolresdr Chemistry. The Inaitole for Cuscer Reee.rcb. Pbile- dclpbie. SOUTI/WEST RESEARCH INSTI- TUl l" Se. Antonio. Tet. DAVID M. SPAIN. M1), D.•enor, Dr- r..rmrwt of Pathology. lbe Rrookdele li.sptt.l Center, !(uoklyn, N. Y. I ALF.XANDER SPOCK, M.D.. Assistant t'ro(rssor of Pediatrics. Duke Utdver- eNy Medical Center. Durb.re,N.C. FREDERICK 1. STARE. M.D. hofrs- sor of Nrrrltiow. Huvud lJniversity School of Public He.lth. Boston. C. HAROLD STEFFEE, M.D.. Director of Lifloraro.ies, Methodist Hospit•1, Memphis. JACK P. STRONG. MD. Associate Professor of Pathology. LouisianeSlNe University School of Medicine, New Ork.w. MARION 1. SUIZIEROER, MD., h+- /rssor and CAaira.rr. Derartwsrwr of Dereratology and Syph y, New York UsdverehP-Rellevu. M ' Cet ter, New York. RENATO TAGIURI. Pu D. Assoei.q hofrssr of tsTcholop, Oredu.N School of Rutlncr Ad.sinistr.Uoa. Harverd U.i.ereity. iodow. MARC D. THAMIM MD. Stdor Rr- srrch Frl/or, M.~o Clinic snd Fow- 4Uork Rochester. Mir. CAROLINE REDELL THOMAS. M.D. rrofessor En.rri)rs of Aledklwe, The /obos Ilookins University School of Mediciwe, DeBirnore. JEROME F. THOMAS. Pu.D. t+ro essr of Sanitr) EnjiwrerisR. U r of Cali/orni., Berkeley. )AMPS E. P. TOMAN, PMD. rrolrr aor and Chdrwww, DrParrsnent of h.rl twoco/q l, Chicago Medka/ School. I.- s(MWe /a Medical Reseercl, Chicago. JANET TRAVEI.4 M D. AssocYN Professor ol Cl/wkd L+hrsiwcologr, Cornell University Medical Coaep. New York. 1.1E SHA TSAI. hr D., Resrrch Asso- clre Jw Pathology Ydt University School o/ Medicine, Wcw H.ve0. Conw D. M. Ti/RNER, Ps(.D., Head. Drpart- rwrnt of Drug lifrraiolistw and Iio- ehanbrry, 1lsileton L.bw.tories Fu- rope, l.td., Il.rropte, Yorkshire. Eng- lend. UNIVY.RSITY OF SAN FRANCISC'O, San Francisco. 101 UNIVERSITY OF SOUTHERN CAU- FORNIA, Lot Angeles. ELI.IOTS. VPSF-LL, M.D.lrofrssrawd Chairman. Department of rherwrecof- of7. Pewnsylvsw S(.1e U.ivenMy Cd- Medidwe, Milton S. HsnMy ZZ Ccaler, HereMr. BRANISLAV VIDIC, D.1.. rrofraeor oJ Anetora,y. Georgetown Utra.ereNy Schools of Mudiei.e and De.tisuy, W..RIttRtork D.C. ROMEO A. VIDONE, M.D, Assodw Professor of rrholo4r, Yale Usilver- eNr School of Mediun., New HeveR Cows. PETL'R K. VOOT, Pst.D, rrofrssor of Mkrollotollr. University of We.Yift- Ion School of Medieine, Seattle. E. D. WARNER. M.D., Professor of r. rholoRy. Suie University of Iowa Col- k!e of Medicine. Iowa City. SHIPa.DS WARREN. M.D., Director d Laboratories Cancer Research wu(-1 r(w~. e.~~ EEnRl.nd Dtecone. Hoqi- YASUSHI WATANASE, PM D. Asrr elNe Mrmbrr, The Wisl.r IastlluN of Atauar.y and Biology. PRUeddofl+.. BARBARA K. WATSON. Pw.D. Aadr- a.r /acrtrblotlu, Me.s.AueeUs O»- rd Hos011.1• Research Associate M lacterblo~y ~wd f.nmrswlop, Her- •srd Mcdtcs+t School. Roeto.. JOHN S. WAUGH, Pst.D., rrofessr o( Chrmiury. Massachusetts lrarit ofirte TccR.o/oS1, Cambridge. RICHARD L. WP.CIISI.F.R, M.D., Ci'lo- /r.f t'hys/alogiu, Mode/lor. Ho.pllel ItwiNMe of Reseerck Pitub(wp. JOHN V. W!°Il., MD, Asds/awr t'ro- lrswr of Mrdkiwe. University of C.do- rsdu Medical Center. Ikaver. A. WFINST()CK, Pw D., Research Ib- rhrmist, LJfr Srirnres D/rislow, IfT Research Iwstitute, Chicago. RUSSEtI.L W. WFLI NR, M D, t•athob otrst, Memorial Hospital of ('Lcna Counly, West ('bester, P..

l ! r . A. STANLEY WELTMAN. PH.D. Aa.a clue Profeuor of PArwr.coforr .w/ Reie..cA, broatly. College of M.r- encr, broo"yR N. Y. SIMON N. WENDER, M.D. ReurcA rro[tiior of /iocAewd.t.y. Urr1.e»Ny of (A/eAore., No.m.a DUANE 0. WENiEL, M.D., rroles.w .wf CAo/.www, Dep.rM.ewf oJ f•Ariw.. c y owI ToikoJq )p U71. of .a... School ot ~F Lw re.ac- THOMAS C. WESTPALL. h.D.Frofem- .or oJ PA.rwrcolos). U.itenily d Vir- {iwi. Sc6ool of MedicMe, C6rlww- •ilk. FREDERICK F.. WIIISKIN. M.D. C-M, Dirccro., Dl.idon of Hee1rA e+wl Per- .owol/iy E.rU/M/un., TLe Ar Cealp of New Ey1e.d. lee, doros+. ROOER l. WILUAMS, M.D.. ho(ea.ew jCAeiw/ur,; Director. Cleyro. ~-orw- ow .bcAcrwk.f fwalrrre. The Uel- .er.hy of Tes.e. Au.tin. DANIEL H. WISEMAN. M.D.. AuLf- ra rrofcuor of Pedi.nk+. Uai.ereill of Sowber. Cdifa.wl.; Childreo i Di- ti.io., La Angeles Couely Oe.er.1 HoMM.1, Los Angeles. I 1. EDWIN WOOD. M.D_ le.rrwcrar iM Me/klwr, RoNo4 U.Irereily Sc6ooi af MedklMe, soatow• SUMNER WOOD, Irr. M.D. Au(u.wr ho/euw of tuAoJojy'. Tbe lof/ti Ha"Me U.i.a.iq Scbol of MeAI- eMw \ehi.~or.. /OHN P. WYATT. M.D.. r-o,euow of rrAo1 osy. S.iet Ladk t).i.cr.kr Scbo1 of M.didn., St. Lc+uk 102 I I I INDEX OF PRIN(VAL IIW,E.S77GATOR9 Abood,LO..s1.s2,s3 Arcos. 1. C.. 7.8.9.10.11.12 Seoedicl, W. F-,17.18.19 Sieg, R.1..43. 44 Ilroob,R-I',26 Ch.lon, l., 25.45.46 Coc6r.oe; C. O-. 47 Coliee„ A. s., 31.12 Costs. P. T.. )r., 81. 82 Priserg. L, 77 Friedmaa, O. D.. 90 Oieks, l. B-.1 3 Oanod, l. W-, 67, 6~ Ou.reeri, l. l., 28 Ouir1li., II. A.. 21 H.moaA, M., 33.34 Hencowitl4 H. s., 75 Io.cAirn, li. L.. 77 1.eoQ, A.. 28 Jusko. W.1., 63.64 Kouri, R• E, 22.21 Kulliuder, S.. 76 Leele, E.. 64.65 Lynch. H. T.. 20.21 i M.rB., R- R-,14.15.71 McKea.i., H.. )r., 66 Meia, H.. 24 MNebeN, l. A., 69 Oeacl, P.,19 rieroe, C. W.. 75 R..1...10,1-, 78, 7! Rrco, M. A.,16 Reid. L. 27 Rey.old., H. Y., 70 Rosecr.as, l. A.. 49, 30 Ryr, U. S-, 36.37. 32. 39, 40.41 S.Mey, D. V. R.,14, 33, 36, 37. 38, 39 ScAmid1,1., 59.60.61.62 Solot, L. A., 41, 42 T.1mw, D .W., 74 Th.ana, M. D•, 48 Tr.d.,1.,33,4t Unamra, E- R., 72.73 Weinbwnr, O., 29, 30, 31 WAilmire. C. E-, 24 Wip,1. A., 34.35. 36 WoK, O., 17 ZeWea, S. O., 62 .103

INDEX OF SENIOR AUTHORS Abood. l. c., 32. 5 3 Abrams, W. R., 31 Arcos,l. ('., 7.9 Artus, M. F., 9, 10 Aronstam, R. S. 31, 52 Astedl, B . 76 Bedigian. H. (i .24 Belkr, t). 1., 73 Benedict. W. F., 17, 16 Brooks, R. E., 26 Bryanl. (3. M., • Card. l. P., 69 ('arp, lt., 29 Catta,l'. E., 65 ('ederl8l, R.. 77 Chaton, 1., 25 ('hance, W. T., 50 (halurvedi, A. K., 37 Christner, P.. 30 Cohen. A. B., 31, 32 Costa. P. T.. lr., 01 ('owan, t). A.. 67 Daks, l.. O., 80 Damini, 1.. A , 66 Friedman-Mor, Z.. 45. 46 Uorrod, l. W.. 67, 6!t (luarneri,l. 1., 28 (3uirgis,11. A.. 21 (3uslow, E., 41 Hagen-Cook, K., 17 Ilammer, R. E., 69 Hamosh. M., 33, 34 Hart. M. A., Ji 1lendenow, 0.1., 56 Ilernaada-V.aquez, A., 34, 35 Hunt. S.. 39, 60.61 lohraoe. D.. 33 /onea, P. A., 19 lones, R., 27 lusko, W.1., 61 Kaprio,l., 75.79 Kirn, K.1., 75 Kouri. R. E., 22. 23 Kunj,1. T-, 74 lawtence, E. C., 71 l.eete, E., 64 Lynch. H. T.. 20 McCrae, R. R., 82 McKennis, H., )r., 66 Mcl.emore, T. t.., 14. 15 Ochilb, R., 59.59 Oesch, F-, 19 aub.dewo, l. 0.,14 Pahsch, E., 77 Raaco, M. A., 16 Revak. S. D., 47 Reynold., H. Y., 70 Rose,1. Q., 64 Ro.ecrata,l. A.. 49 Rowelt, P. P., 54. 57 Ryan, l. W., 36. 39, 40, 4 jr Ryan. U. S.. 37, 3 ~1 Sarsna.l. S. M.. 43. 44 Sarna, S.. 79 Saslry, B. V. R., 55 SchecMer, N., 61 SoloA, l.. A.. 42 Sladecker, M.1.. 72 Tadakuma, T., 75 lAames, M. D., 48 Travis, l., 48 Unanue, E. R., 73 Van Canlforl,1., 13 V arma, K. O., 42 WanS, (3-K., 62 Warr, 0. M., 71 Weinsaum, O., 29 Weiahaar, R., 44 Wharton, l., 36 Whilmire, C. E.. 24 Wi`Sins. R. C.,47 Woo. Y-T., 11, 12 Z.e/dea, S. (7., 62