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ANDREWS 0. YiCE PRODUCTS CAPITOL HEIGHTS, MD lK/ i

1980 REPORT oJ THE COUNCIL FOR TOBACCO RE3EARCII-U.3.A., Ire. TRE COUNCIL F()R TI11tA/'f:O ItF.Ct:Altfil-I/.c.A., lne. 110 F:.N 591h 11ree1, New Y..rk, N.Y. 10022

SCIENTIFIC ADVISORY BOARD to The Council lor Tobacco Research-U.S. A., Inc. as ot December 31. 1980 SHELDON C. SOMMFRS, M D., CA.Mwrw Dlnector of Loho•ato.kr. Leaos 102 Ila.pitai Cf/nkd ho%ssor o/ taAofory Co1k~e oI Phrsicians ! Surgear d CohnnDia Univenity New Yatt, New Yort RICHARD 1. B1NO. M.D. Dhedar of Cardiolngy and fntnrwa.l Mrdkier Hantington Memorial Ho.pitai, Pasadena, California Pbolessoi of Medieine Unhersitr of Southern Caiitornia School of Medicine Let Angeks, Cali(ornia ROSWELL K. BOUTWELL. Px.D. Peo%ssae of t7nrubtsy McArdk Laboratory lor Cancer Research thniversky of Wisconsin Madison; Wisconsin JOSEPH D. FELDMAN. M.D. Head, DepaHmettt of Immm"thobn Sa4" (lWe and Research Fotn+dation L.a loila. CalKornia Wi1.UAM U. GARDNER. Ptt.D. Sdentl/k DJrecao.. The Conncil for Tobacco Research-U. S.A., lax. Q. K. Haart Professor of Anetorny (enreritw) Yale Udvenit' School of Mcdicine New Have., C.rrrectkut ROBERT 1. tiUEBNER. M D. Chief. I aborstory of RNA Tumor Viruses National Cancer Institute Bethesda. Maryland i.EON O. IACOBSON, M D. loseph ReRenstein Pro/essor of Biolnairof Scienres i )niversity of Chicago I HENRY T. i.YNCH, M.D. rro%sso, and Cbee.ewn Department of Preventive Medicine and Public lleaith Creighton University School of Mcdicinc Omaha. Nebraska HANS MEIER. D.V.M.. Dr. Med. Vet., M.R.S.H. Senior Stel Sekwt/st The Jackson L.boratory Bar Harbor, Maine GORDON tt. SATO. Ptt.D. Pro/rssor of AtoWgy tlniversity o/ Calilornia, San Diego 1.8 Jolla. California Se1..liie $tat.f Tit. Cr.we/i WILUAM U. OARDNER, Ptt.D. Scfenft/k Dbscto. ROdERTC. HOCKE'fT, Pw.D. ReserneM Wrrclo. DONALD H. FORD. Ptt.D. V1NCL'M F. LISANTI, D.MD. Anociare ResreoswA iNrerctot " Asisdrtt Rssu.Y# Db.efer DAVID STONB, PM.D. Associare Resercb Direna.

CONTF.NTS Introduction Abstracts of Reports . . . . . . . . . . . . . . . . 7 Cancer -Related Studies . . . . . . . . . . . . . 7 The Respiratory System . . . . . . . . . . . . 29 lk.rt and Circulation . . . . . . . . . . . . . 41 Neuropharmacology tnd Pltpiologr . . . . . . . . . 49 Pharmacology .nd Biochemistrr . . . . . . . . . . 57 Immunology and Adaptive Mechanisnu . . . . . . . 69 Fpidemiology . . . . . . . . . . . . . . . . EO Active Projects . ... . . . . . . . . . . . . . . . 90 Completed Projects . . . . . . . . . . . . . . . . 99 Inde: o( Pri.cip.i lavestigaton . . . . . . . . . . . . 111 Inde: of Senior Authors . . . . . . . . . . . . . . 112 Int: oduction Since iis formatioe 27 years ago, the Scientific Advisory Board so The Council fw Tobacco Research•U.S.A., inc., ha reviewed many formai awd i.- /ormal applications for research support. The direct resnk is that 413 diAereM projects have been /uwded by The Council. based upon the reeanmendNios of the Advisory tloard, 1o supporl investiptors in 2S9 medical schooh, hospwah .nd research institutrons. Each ol these projMs, whether active or eoanpleled, is listed in Ihis year's Report. Total fwwlinR through 1900 was f61.00(1.000. The chief output of The Cornci/'s tln.wei.l support a.e 1.862 scieMiOe papers and reponsl published by drese invesliwors in rwany diAerenl medical and scientillc journ.b, covering a wide range of srbjeets. The publicatiow of these research resnNs .dds to the sciaNilk literature a.d to the stoehpik of information from which, it is hoped. will come .wswen so cornpks proMewr such as those associated with uncer, cardiovascular diseases and puinrow.ry ailments. Abstracts of wunuscripls published during 1980 that actnowledRed Council support comprise a sectiow of tbi. Mww.l Reporl. During 19tl0 two new wremhen were added to the ScieMifie Advisory Board to provide additional eapertise to that distinguished body. They were Roswelf K. Roarwell. Ph.D.. Professor of Oncology at Ihe MeA.dk Laboratory for Cancer Research, University of Wiseorrin. Madison, and Gordon H. S.Io, Ph.D., Prolessor of Biology. Universily of California. San Diego. I 5

Abstracts og Reports I Following are aMltacls, approved by the .ulhors, of reports on .ew research aclnowkdtint support Irom The Council that have appeared in scirw IiBc pwrnsls since publicalion of the 1979 Report. The name of 1he recipient is in italics. lhe ahuracls are Rrouped under these headings: 1. Cancer-Relaled Sludia, 1'. The Respiratory System. Ill. Heart and ('ireulalion. IV. Neuropharmacobpr and Phrsiologr. V. Pharmscobpr and Siochemisuy, VI. Immunolbay and Adaptive Mechanisms, VII. Fpidemiology. 1. C.wcor-Rot.t.d Stssdl.. ARYI. IIYDROCARSON HYDROXYLASE IN NORMAL AND CANCER POPULATIONS The roks of aryl hydrocarhon hydrosylne (AHII) and its inducibility in polpcyclic aromatic hydrocareon earciwoRewesis ue discussed at length in what is essentially a review of the research iw this .re.. With respect b Iwww AIIII, /his poup's cwrenl .iews are sunwn.rittd as follows: ( I/ the enrlnrte's inducibility diRen among variow Mdi.iduab; (2) this di/lere.p is pe.NicaNy con/rolh:d; ( 3) the distribution plterw of ANN Iwdnehy in the normal pnpolation may merely reAecl d.y-lo-day vari.Ailily In the lymphocyte arslewt, but it also suggests polygenic eoaMrd of Mte enzyme system; snd (4) lrsy and orophsryn6eal cancer patients have a AiRlter A/IH inducibility tw Mtl found in /he normal population. Indications are. however, thal cancer per ar does um cause high ANII inJrcibilNy. In plients with other types e( wNR- nancies. the enzyme inducibility was indialiwRttishab/e from that found i. the mKmal population. Studies Ihm show aw iwcre.sed iwcide.ce of hydrocarbo.- induced wmors In animals .uscepible so ANN irduction, harever, stNResl that such inducibility i. a eriliral factor M•1A. oecnrrence of chewtic.llp W duced turnun. Never/hdess. Ihe deve/opwrcnl of a more rdiable melhod of . assessing AI/II br/ucihiliMr in hwmns is Orersyuisile to Ihe es/aMisMee.1 ef this factor's role as a determinant of wsceptibilitr 1o hydrocarbon-LAuoed lumors in man. Arnwr, M. S.. Yamuchi. T. and lohson. D. A. In: Grifiin, A. C. and Shaw, C. R. (eds-): CwriwoKewsr /denti/k.rion .nd Mecbuniuns o/ Action. New Yorh! Raven Press; 1979, pp. 113-136. Oth.r swppert: National Institute of General Medical Sciences and Nalio.al Cancer Institute. From the Ikpartmcnls of flioW.ty and Sioma/hematics, The Univrrsily of Teass System Cancer ('enter, M 1). Andersoa /lospital and Tumor Inailuu; and the Umvcrsity ol Fesas Mcdical School at Houston. 7

ARYI. FIYf)ROCARBON NY1)ROXYLASE IN IIUMAN CANCER POPULAIIONS lhe genetics of aryl hydrocarbon hydrosylase (AIIN) inducihility Rod tumor suscepribilify have been sludred eatensively in inbred mice, and some of the eaperrrnental animal models have sugjesled that AHII irMlucibilAy could be an important delerminanl of susceptibility so hydrocarbon-induced Cumors in humans. 10 lest this hypothesis. AHH activity was measured in cultured human lymphocytes. One rnvestrelNor, swveyinR inducibility in a poprlaliun of 35) healthy donors, repwted that it (eN into three groups: k+w, intermedi- ate and high. lhn trimodal distributio. suggested that two slkks at a sin6k locus were responsible for the genetic control of AHN inducibility. Family uudies were consistent with this eowcepl. The present invesliptors measured AII/I inducibility in lung cancer patients swd palierws with other Iypes of cancer, snd developed a sophinicded, consppMerked syNem for information rettieval pertaining to normal dowors and ea.oer palienu. Over 700 normal d.+nots and 600 cancer patrems, inc/udinR approaimalely 250 with lung can- cer, have been tested in this system to date. The studies reported here lead to the following summar7 of A11H induabilily in humans: (1) individuals difler in therr AIIII n+ducrbdry; (2) the inducibilily differences obsersed be- tween prople are under genetic comrol; ()) the rnimodal distribution o( AI1/1 inducdwlity observed in the normal population may stem in p.rt from day-to- dar variability in the lymphocyte syuem, bul aho suggests poyflenic cawrd of the enzyme sysuem; and (4) lung and oropharyngeal cancer patients ea- hrbrl hrgher A1111 rnducrAMhty than the normal population. No cause and eReet relationship can he rnierred from these studres alone, but viewed in the contesl of what has been shcwn clearly in e.Perimenul anmmsls, the human lymphocyte A1111 uudret suMe.t that AFl/l rnducrhdd7 rs an imlrrrlant de- lermmanl of suscepahday to hydrocarbon induced tumors Arnorr. M S. Yamauchr, T and /ohnsron, 1) A /n: /ones, P. W. and 1 eber, P(eds 1: Polrnrt lear Aromatic HydrorarAont, Ann Arbor Science Publishers. Inc , 1979, pp. 779 791. Other s.rp.rlt National Inslitule of General Medical Sciences and National Cancer Institute. From the Departments of Biology and Biomathemalics, The University of Teaas System Cancer Center, M.D. Anderson Hospital and Tumor InuNute; and The University of Tesas 1leshh Science Cenler, Medical School a1 Houston. ARYI. HYDROCARBON IIYDROXYLASE ACTIVITY IN PUI.M()NARY ALVEOLAR MACROPNAO!'S AND LYMPHOCYTFS FROM I.UN(3 CAN('ER AND NONCANCER PATIEN7S: A CORRELA'I ION WITII FAMILY IIISTORIES OF CANCER In this attempt to determine whether differences in aryl hydrocarbon hydrosylase (A/lll) irducrbdrty esist between normal and lung cancer pa- uenrs. A1111 actuvNy was maasured in pulmonary alveolar n.actoPhages (PAMt) and prrrplwral hio.rt ItimpMrcyies from cigarette smoken with and without primary lung cancer. The dals allowed comparisons between 1ht nor- rnal and lung cancer patients for 26 variables, including sile, occupalion, aes, and family histoties of disease. Frequency dislribtMion analysis of AHH i.- duclion ratios for the two groups revealed as increased aumber of iwdividtals in the lung cancer patient liroup with high lymphocyte induetiow values. Sratil an increase was not shown fot hi"AM AHH values in hrsg cancer pslie.b. When individual PAM and lymphocyte AHN values wen compared belwee" noocancer and lung cancer patierws, there was a positive eorrdslion for now- cancer patiems, but /hese values did no1 correlate (or hswg eaacer paliewu. These cancer paiieMS were divided into three subgroups dsowinR (•1) Msli PAM and low lymphocyte AIIII kvels, (/11 low PAM awd low lymphocpte AIIII kvels, and (111) low PAM ard high ymphoeyu AHH kvels. Whew the incidence of family history of eawcer was compared (or lhne•subporps, none was recorded /or persons in srARroup 11; however, individuals in sub- groups I and 111 presented family cancer history iwcideaces of 9.5% and )9.1%, respectively. Patients in group 1/1 averaged sia years younger Ihr those in group 1. These dats support the hypothesis that high levels o/ AI/N might be associated with increased levels of hrwR cancer In rnat, and Ihey indicate that, for the sn.ysis of A1111 levels in hrwg eancer palients. Yse of the cultured lymphocyte system as a sole indicator of high All/l levels ia .o1 warranted. These data sho indkale that genetic facton are espressed In the initiation of lung csacer. Mct.enwre. T. 1.., Alarrin, R. R.. Springer. R. R., Wrsy, N.. CantreR, fd. T., srd Susbee, D. L. Rloc/iemk./ Cenerks 17(9/ 10):79S-fi0S, 1979. Otii.r auppor/: National InWilules of Health, National Cancer Irrtitr/e, Americsw Cancer Sociely, awd /he Veleraru Aderinistraliow Hospital. HorrNO.. From the Department of Mediciwe, faaplor College of Medicine. How1o.; Veterans Administration Hospital. Houalon; Deparlnsewr of Pharrnacolop, Teaas ('olkge of Osteopsthie Medicine. Fort Worth; awd rhe Deparlnse.l d Biological Sciences and the Genetics CesNer, Nor1h Teaas S1a1e University. Deelon. I DOSE-RF_SPONSE RELATIONSHIP OF RAT ARYI- HYDROCARfaON IIYDROXYI.ASE AND EPOXIDE HYDRATASE INDUCTION The data reported here sapport 1he hrpolhesii thN different regalatory mechanisms control sryl hydrocarbon hydrosylase ( All H ) and eposide hydrNw (EN) activity and that h selective inducer o/ liver entyates, auch as pheno- barbilal, may also affect the inducibility of estraAepalie monooarflews.ea by other chemicals. The Allll in ra1 h" and tidney is highly sewsilive so I.- ducers in cigarette snake and smole cordcraale, wfsile EH activity is na1 a/- (ected by these agents. Both AINI and EN can be indreed selectively is pri- rwary rat liver cells in culune. I.ow doses of beaw(.Isnlhracene (SA) prefer- entially enhance Allll aclivNy, while rrans-sliRsene oaide (ISO) and vuiorrs e 9

.~..1. .~f . . anriosidants affect only the F11 activity. Phenobarbital, which potentiales in- duction of lung AIIH activity by low doses of benro(.)pyrene, also induces AI111 activity in cell culture and has a greater than additive effect when mised into the culture nxdoum with BA. The effect of phenobarbital on A1111 induc- lion is M.K\cd by TSO but not by the ioducing effect of BA. These results suggest that phenobarbital is not merely an A1111 inducer, but also has the capacity to potentiate the action of cheereals that belong to a direrent class of smhrcen. In parlicrrlar, they derrwnstrMe that phenobarbitsl a/one induces  cylochrorne P-ISO linked A1111 good causes the accumulalion of rnore cyto- chrome P,-450 associated enzyme whew rrrised with BA. CirGn, 1. E. rr .1. ArrAivr, o/ Torknloty Suppl. ):20l-216, 1980. From the I.aboratoire de Chimie MEdkak, Instilut de Pa/hologic. Univenilf de I rtje, I ii`le. Belgium. COMPARItON OF ARYI HYDROCARBON HYDROXYI.ASE AND FPOX11)E IIYDRATASE INI)UC"IION IN PRIMARY FETAL RAT LIVER CI't.t. ('UI TURE The effects of benz/alanthraceae (BA). phenobarbital (PB). cigarette smoke condensate (('S('i, 2.).7,t.tetrachlorodibenzo-pdiosin (1('DD) and rr.nr-stilbene oside (TSO) on aryl hydrocarbon hydrosylase (Alllli and eposide hydralase ( Elf ) activity have been studied in primary /etsl rat liver cell culture. The two enzymes vary iw their response to lhese chemicals as shown by the following: (1) the concentration of BA. PB and CSC required /o induce Allll is lower than the one treedcd for signi/kanl irrduction o/ El/; (2) TCDD and low concentrations of A aekctively induce AHII; (3) with BA. PB and CSC. the ANN induction kinetics are not the same ns those of EH itduction; (1) TSO ieduces EN selectively. In the early phases of EH induction, RNA and protein synthesis r well as the eontimrous presence of the ir.ducer are required. When EN activity reaches a plateau, neither iaact RNA uor protein synthesis is required 1o tnaiMaiw enzyme activity at its opti- mal level. The Ell acti.ity decays with the removal of the inducer, and its biololic haN-Hle Is eslimaled to be abow 72 hours as opposed 1o about 10 horrrs for A1111. While TSO prevents the irduction of A1111 by PB, is does nol block that twcdialed by BA and CSC; but added together wah PB, BA, CSC or PB plus BA, it Induces the F.H activity in a more than additive man- t.er. TUis eflect, however, is only seen after sia days of continuous Ireatmcm. According to thesc results, in this particular timre eultute model, the mech- anism of Alll/ and 1'.1/ induction can clearly be dissociated. (Ioujow, F M, V P-n Canllort, 1, and Cklrw, /. E. ('A....k.. N..J.•rr..1lwILw/N.nr 1? M1 175, 19RQ I r.•n. 11v 1.r....I ..... .k 1 h.r..r Mtdrak t~ lu.uulu~~e, In.l.w~ Jr Pa~hoN~~~e, (1n..e..,i1 .1. I 1 r. 1.1 p M./r.um i I CORRI;LATION OF INDUCIBII.ITY OF ARYI. RIYDROCARBON IIYI)ROXYt.ASI: WITH SUSCEPT1B11.ITY TO 7-METHYI.CIIOLAN- 111RENI:-tNDUCNb LUNG CANCERS Intratrached treatment with polycyclic aromatic hydrocarborts may iw- duce specific genetic regulation of AHH adi.ity. Previously tned methods for producieg lung carcinomas in inbred atrains of mice wuesl a possible aaimal model in which the susceptibility to pulmonary carcinomas may be specifieaNy linked to the capacity of that organ to wrelabolite chemical earcinoRens. Iw Ihe system described here, mice of the CS7BIJ6Cum. DBA/2Cum, firsl flli. (F,)Snd bactcrots progeny from the two parental strains were evaluated /or susceptibility to )-nrethykholanthrene (MCA)-induced lung cancers. In /he crosscs, AI111 responsiveness segregated as a single wlowmal dominant aene (A/i locus) The AHII responsive mice esprened a very much higher A/IN activity/g wet weight liver alter intraperiloneal treatment wwh MCA tham the nonresponsive animals. /ntratraeheal administration ol M(-A (four 500 ang dores, a1 weekly intervals) caused a.ariely of pulmonary malignancies among the mict surviving one year after IreNtnent, inchding sauamous cell earci- nomas, alveolar adenocarcirwmas and adenosqwmous cell carcinomas. The A11H-responsive CS7BL/(iCum, F, and C37BL/6Cum a F, mice were wweh more susceppibk 1o MCA-induced lung eancen than the nonresponsive DBA/ 2(•ums. Furtherrrw.re, /he lung cancers found in DBA/?('wn a Fr bactcrors progeny did not occur randumly, since aipwOeantly more tumors were found in the AIIll responsive offspring rhan in the nonresponsive ones. Thesa data knd lo support the eaistence a/ a genetic link between the capacity to respond to polycyclic aromatic hydrocarbons through increased levels of AIIN activity and susceptibility to certaiw types oi maliRnaweie.. Indications are /hst aa- ceptibiliq to MCA-induced WnB earcinorwas is related 1o the AM allele. Ttieae results also suggest that animal modeb i.volviwg 0bred strairu could be trrd to study the control and repdMiow of epMhelidly detived tumora, tha lype moal /reaue.tly observed in the hurw.. Iwtg. Kour:, R. E. rr d. (Afkro6lolopkd Asrod.res, lnr.) Cancer Letters 9:277-2t1, 1980. Other w'poorfr National Cancer IratitWe. From the Departmewl of Biochemical Oncology and Department of Eapt:ri- mental Oncology. Microbiological MtiociMes, lr.e., Belhesda, Md. XI?NOIROPIC VIRUS EXPRESSION AND SIISCEPTIBII./TY TO )• Mt 111 Yl.(-11OB.AN I IIRE N!:-INDUCED CANCER Mice front two inbred urains. NZS/SI.NI and 129/1, nrd their variar genetic crosses were studied here lor evidence ol a genetic linlage betwass spon/anous production of in/ecri«ws aenwropie (X-tropic) virus aad auscsF tibility to chemically Induced cancers. The pantnlal alrawn and Fo, bactcror, and F, progeny bNween thcse two strains were partially spknectomized to M- oertain X-tropic viral status and were wbsequenNy treated .ubculantously, wwith SOtI r6 of ) melhykM.lanthrenc in tri.rctanoin. Progeny /rom second bact- erosses were also le.tcd lor Iheir X•Iropic viral status and suscepibility to l-nKthylcholanlhrcne carcum.gcnctis. All mice were obacrved (or evidence of 11

fibrosarcomas at the site of inoculatioa over a 10-momh period. In order to specifically address the question of she role of X-tropic virus esprcssion on susceplibility to chemical carcrnotenesis, populNiotss in which vwus espressNon sepega/es were suudsed llere, the carcinogenicity of she chemical was deter- mined by obtaining a single value, the C1, which uuilired both tunusr incidence and latency period. After )-methykhdantbrene treument, 42% of Ihe virus- positive mice developed tunsors r cowtpared to 22% of the virus-nepti.e annnals. Ilowever. the average latency period waa much longer for the virus- positive mice. so that the resuNaat CIY for Ibe two populations were quite similar. Overall, the resuMs seew here suggest that Ii1tk, if any, correlation esists between virus espression and 9-wntbykholanthrene earcino6enesis, and that there is no densonstrabk genetic linkage belween the Iwo. Nayar, K. T., Levy, /. A, O'NeiR, B., aa/ Kowl. R. E. (MicrobldoCk.l Asroci.res, Inc.) l'.ncer Reree.rA 40.1161-1)67, 1980. From (homet Technologies, luc., and tM Department of Siochemical (M- coto6y, Microbiological Associates, (ne., Oetheda, Md.; and Cancer Rcoearch Institute, Uaavenity of Cah/ornia School of Medkine, S.e Francisco. COMPARATIVE FFFE(-'TS OF INDOLE AND AMINOACETONITRII.E DERIVA7IVFS ON 1)IMETIIYI.NITROSAMINE-DEMETNYLASE AND ARYI. HYI)ROCARBON IIYDROXYLASE ACTIVITIES Certain substances such as the 7-substNuled indole derivalives found in crucrferous yegetabks, including bru..el spouts, cabbage and eaulsllower, alter the biological responses to chemical eucino6eos. Of these eompounds, which induce aryl hydrocarAors hydrotylase (AIiH) and inhibN polycyclic hydro- casbon•induced neoplasia, at knt one, )•indol7methanol, she induces amino- pyrine Ndcsnethylase and p-nilroaniaok Odemethylase. Indole itself also in- ducts dimethylnilrossmine (DMN)-&netbylass I in the rN, as does erypto- phan. Mnino.cetonitrik, a nsiaed-Hwtdion osidase modi/kr, iMerteres with DMN ntetabolisrn, inhibib DMN-induced rnethylation of Aepatic RNA and breakage of DNA, and protects a#aiwsl DMN-produced injury to micsosomal amino acid incorporation; it she protects against DMN-induced disorRani:a- tion of hepalk fine structure and DMN-i.duoed hepatic eareinosenesis. Here. she eRecss of indok and amino.utonitrik derivatives on the two enrymk forms of DMNdemethylase and on A/I/l were studied in viro. Indole, ) i.- dolymethand, 1-irdulylaceloniuile, )-indolylatetone and 1.•lrypksphan in- creased A11/1 induclion l- to 6 fold, but p•)-iwddyk(hned had no e/fect. In fact, Me latter deerea+ed she tissue endoplasmic reticufum contenl (/e., micro- somal protein/unit weight) by 21%. Only I:tryplophan induced DMPI-deme- thylase I and only it and )-indolymethsnol induced DMNdemethylase 11, about doubling the entyme activity in all three instances. Anwta.cetonitrik strongly repressed DMNdemethylaae 1. Substitution of the .rnino group greatly decreased or abolished mised•function osidase repressor activity. For esampk, imimdiacetonnrik displayed only ahout one•R1th the repressor ac- tivity of the parent cnmpiwnd, while nitrdolriaeetonitnk and dimethylamino- acetomude seemed to he inactive Ncither the derivatives studied twr <he par- ent comlwund, huwever, had any eflect on 1)MN-demethylase 11 or AtIH activities. Since these mised-function osidases represent critical steps in the rnetahulnm of DMN and polynuckar hydrocarbon., she modifying eRects of iodok and aminoacetonilrik and their deri.atives demonstrate she potential compksity of assessing she eRects of dietary constituents on the carcirakc.k responses. It is suggested that such constituents can generally aber ausceptrbilay to chemical carcinogens and in some innances, can further amplify the car- cinogenic responses to one or nare combination of agents. Arcos, !. C. rt d. C.ncer Leneri 9:161-167, 1990. Ot16.r a.'prt r IloRma.a-La Roche, Inc. From the Seamen's Memorial Research Laboratory, U. S. Public Health Service Ilospitd; and she Depa.ement of Medkine, Tula.e University Medical CeMer, New (Jrkans. 111(i11-PRESSURE LIQUID CHROMATOORAPHIC ANALYSIS OF flEN2O(a)PYRENE METABOLISM BY HUMAN LYMPHOCYTES FROM U()NORS OF DIFFI.RENT ARYL HYDOCARBON HY- DROXYLASE INDUCHlILITY AND ANTIPYRINE HALF-LIVES Lymphocrtes from sis hunun donors were evaluated by meaes of bi6h- pressure Ipud cbromatography (HPLC) for tbeir ability so wtetabolin bewo(.)pyrene (flP). In these subjects, tbe aryl ttydrocubow bfdroaylna (AIIH) irducibility ratio ranged from 2.4 to 1.6 and the aaipyrine plnrraa hdf-IJe from eight l0 17 bows. Whik the results show that f!P metabolizing activity does not diRer qralMMively amowR hwan dowors, there .re qtrrlMa- tive variations in the isducibility of SP wrctabolites, reReNi.s known individud differences in AIIH inducibility. Several fIP metabd'stes were iaeaiRcd: 7,f1- drhydrodiol, quinones, and 9-brdroar and 3-ttydrosy phenoh. The HPLC data for the induction of SP phenol prodw.yitM oenelaled well witA she kwwo A/ill inducibility ratios for she dowors, as delernsined by the tosveMiow.l fAwromttrk AIIIf assay. These stssdies she indicated that the induction of bnuo(.)pyrene-7,tdibydrodiol, proposed a/he proairaate carcinogenic foew of flP, does not parallel RP phenol indueliow. 1s addition, there was a s1ro.R negative correlation between AIIH inducibility and measuremeas of p/astw4 antipyrine or urinary •-bydroayantipyrine half-life. At best 1hen. AHII in- ducibilay can be presumed to be an indea of the capacity to nsetaboliru IP through nsajor, if no/ a8, patbwars, wRRestinR wseyual /lue of BP Mroup/s the various routes. The data also indicate thN at kan one activalin6 iner- nsedute systcm, the formation of 1!1'-7,rdiol, is not directly proportional to AI111 kvels. C aroo. H. L., Vau6ht, 1. 1l., Mdtinelks, A. /., Pai6en, 6, Ceisner, T.. ad 1lolanowska, W. C.nrer Research 10:1 T0S-1)10, 19r0 OtAer swrrort t U. S. Public lleahh Service. From the Deparament of li.perimental Therapeutics and Grace Cancer Drug ('enter, and the Ikpartmcnt of Molecular Biology. Roswell Park Menoorial Institute, BuRalo. 12 11

I ('AR('IN(NiliNlt'l l YO1:HI N/O(o)PYRI Nli ANI) IIIIR-11:EN (117 IfS 1)1 KIVA I IVI:S IN (')11/1('am MI<'f lhe tomorifeenicity ot a selected number of hento(,.)pyrcnc (HP) de- rivatives was inrestotaled in a tunwn model consntin6 of fiMusarcoma indoc- Iwn in suhcuNaneounly injrcted (')11/(Cum mice. lhe It derivalives used were selected on the basis rd their !n rbro and in rirn hialogie aelrvily, as well as o( their prNtnlial foH ehKnlalinK the .rnponance rd HI'-7,K-dwl 9,tlt.cl+oaide in mediating lumor lormati4.n Io1lo.inK subcutaneous HP administration. lhe resulls suRRest that mduction of subcu/aneous fihrosarcomas in thn mouse strain has omportant limitations in spite of SP's ability to cause a high inci- dence of tumnrs. Most signdkantly, she derivalives' polarity appests to mark- edly ahcr Ihew hMAopie aahvity in /hia system Because nrelabolites of poly- eyclic hyJrocarhons are usu./ly more polar than their parent compound, il is ddlicuh to assign a particular one so the a1Nus of proaimate or ultimte car• einnten In additoon, wnh any given eompound, the lurnw 'incidcnce was notably aflecled hy the nature of the solvent. For eaampk, SP's carcirw+teni- cny index decreased about S0•70% when DMSO was used as she vehick rather than trK+ctanoin. (M she other hand, other investilators have found 1h.1 It was almost completely abolished in a/:1 Irioctanoin and beeswax mixture. According Io some previnw reports, tumorigenicity of BP•7,t-diol-9,10- eponide dustereomers on mouse skin also depends on the soWent employed, the highest incidence having been observed with teeltahydrofuran and wilh acetune. Both of these, huwever, proved to he toaic to mice. AddAional re- search may lead to the discovery of a solvent or solvent combination that mini• mirn she efleet of a compound's polarity in determining tumor response. ' Kouri, R- E. .r ./ ( Mir.oAwfotird Assari.rrr, Inc.) 1or.nd o/ the Narioe.J C.nre. lnrrlrrrr 61( )):617-62 ), 1990. From the Depntment of Srochemiwry and Drug MetaAolism, Ilof(mann-l.a- Roche, Inc., Nuwky, N 1; the Department of diochemical Oncology. Micro- biological Associates, Inc.; and the Section of Oxidation Mechanisms. l.aboralory of Ilioor6anic Chemislry, National Institute of Arthritis. Melabolrsm, and 1)iKes- tive Diseases, Oelhesda, Md. SI'CONDARY M1: f ASOLISM OF SENZO(a)PYRI:NE IN I/UMAN CEI.IS While manr mNabotites of benro(.)pyrcne (SP) are known to form in man, receM s/udies have shown thal /hese primary metaholiles are /ransformed further to conjugates of sutfale, glucuronide and gluthalhione. llere, human tymphocytes and lung macrophales were used to study DP metabolism in cancer and noncancer patients. Results oI Ihesc esperiments showed a reeta- Mdoc pro8k similar to that reported by others for human lymphocytes. After hydrolysis of the fractions, the metabolic profiks were slightly altered. The observed data suggest that during a short lime of reaction whese the substrate concentrat«sn is not saturating the enryme compka, a substantial nmown/ of conlugNion occurs via Klucuronide formation lhe phenoh are conjugated to an equal estent wnh glucuronKk and sulfate. Also, in Wng macrophatees the 14 I estent of cunluKation was variable from per.on to person A preliminary aw- vcy u1 livc nom.m.skcrs indKalcd that the extent and selectivity of ineubol- ism was variahlc as well. lhcre was no consistent pattern Io the varialiow in conjugation ol 01' reetahohtes. Overall, the data presented in Ihis paper i.di- cale that cultured human lymphoeytes and pulmonary alveolar teaeropAap have the capacity to metabolire S/' Io several hydroxy and quinone derivatives. lhese arc lurther metabolized 1u polar conjugates, including sulfate and glu- curanwks 1 he proportions of the vanous pimary, seelabolites and eonjuples vary between individuals and vary with the tirne of incub.tioa and eP concta- Iration in a single individual. IMn increasinp and compoundinR the complexity of BP metaM.lism in human tissun. C.ntrerr, E. T'. .r J. rro.reJinRr o/ rh. Wruirn Phwrn.roluey Socirry 22:27)•276, 1979. O/h.r aupp.rlr U. S. Public /leahh Service. From the North leaas Sta1e University /ltahh Sciences Center; Colkge of Osleopathic Mrdicine, Fwt Worth. lea.; and M.D. Anderson Hospital and Tunwr Institute, tlouslon. eF.NZOIoIPYRENE ME1 ABOI.ISM IN RAT FETAL HEPATOCYTES (:UI.TURE. IMPROVED METIIO()OLOaY AND EFFECl' OF SU SSl R ATE CONCENT R Al ION lhe exact rate o( henzn(„byrene (uP) metalsdism in any living syMetn is imponant in that the balance betwcen tht r.riow enzymatic reactions b- volved in this process is highly signi/kanl in deltrmitwn4 the iMtacelWlar ten- centration of the toxic intermedi.tes. Presented Aere :re data on SP tectabolistn in primary letal rat liver cell culture. The tatenl of iw r/ro 1'H)SP eseuboNna was established by measuring all of the tANabolitn retained iw the aeM aa weR as those excreted into the cuMure tnediutw. lAt eaent of the eoajusaioe srd the nature of she conjugates were determined aed the nxtabdite paltcra .aa analyzed by high perlormance liqnid chromatography (HPLC). The fetal hepatacytcs actively metabolize SP and readily e.crele aR the metaboYtes iao the culture medium as wllates and tlucurunide eonjugales, lhe relative pro- pwtion rd the latter varies directly as,a /unetinn of she SP concentration. As shown by the IIPI (' analysis. eP•1,6-quinowe aad -),6-quiaswre are tlle mN.- boldes excreted in she larVit aaounl, suke.ting the probable existence of an active 6 hydra.ylase reactiorn medtanism in the fetal hcpatoeytts Aryl hydro• carlM.n hydroaylase activity nwddkti she overall rate of /!P metabolnm dra- matically but does not a/fecl she qualitative paltern ol Ihe excreted melabolites. Van Cantfurt, I.. (loujon. F. M. and (lirlea,l. E. CArrait'u-Rioluriralln/erurN..nr ?K:117•160• 1979. From the l.aborioaqrc dc ('himic Medreak, lnsutus de Pathobgie. Udversitt de I .ttKe. 1.i2Ke, /irloe/um. IS

I PHA(:O('YTOSIS OF ASS1:STOS FIBERS BY HUMAN PULMONARY ALVEOI-AR MACROPHAGES Ahhough epidemiologic evidence supports a relationship between esporure to asbestoa (AS) fibers and the incidence of cancer, the eaact nwthanism of '•aclion on the human respiratory tract remains unknown. The pulmonary alveolar macrophages (PAMs), which ingest various foreign pirticks that enter the destal tracheobronc/aal ttse, are also the primary line of defense a6ainst inhaled AS fibets, bu1 beeause it M virtually indestruaibk, (his material poses a unique probkm for PAM dnoaiReation. There is evidence, however, that pAa6ocytous followed by eoaling of the panicles with celi memb.ane componeMs, acid mucopdysaceharidn aad hemosiderin convert it to a less tosic form. The present study investipled aome biological iwueraclisns between cultured human PAMs and aewsite AS. Phallocytosis, cytolo.ic.ty and cell surface changes were esamiwed by acannin6 ekclron microscory a/ter the cetli viability had Rrs1 been delermired by the trypan blue dye esclusion method Rcwhs show that low eoncentrations of amwile AS are uny slightly tosic to human PAMs (n vitro. Hipher eoeceMrations, however, cause si6nifkant toskity. Phagocytosis begins immediately upon contact with AS: the PAM either wraps itself around the end of the fiber, ewdocyt'ain6 it by slowly moving toward its opposite end, or w spreads toward each end from the mdline, then completely engulfs it. The macropha6e membranes are also aAected, as re- /kcttd by an increase in cyroplasmie bkbbiag known as :eioais that probably resuhs from Uve ceM's high metabolic activity, and by the appearance of a (iMow-hke martrial in close approswnation with the AS fibers in the area of the PAM membr.ne whcre the initial contact is made. The precise nature of this cellular product is curremly under study to determine its biochemical structure Further research onto these biological interactions may prove valuable in establishing the etiology of AS-related lung disorders. McLemore, T., Coroow, M, Mace, M., Arnmr, AI. S., )enkins, T., Snodgrass, D., liLrrM, R., Wray, N., and Brinkley. B. R. Canrrr L.errers 6:1fi1•192, 1979. Other wFl.rt: Veterans Admioislration HospNd and the National Institutes of HcahA. From the Veterans Adminislratioe Hospital; the Department of tsiology, The University of Tetas System Cancer Center; M.D. Anderson Hoapital and Tumor Institute; and the Departments of Medicine and Cell Biology. Baylor College of Medici.e, Houston. IN VITRO ACTIVATION OF CIGARETTE SMOKE CONDENSATE MATERIAIS TO T/IEIR MUTAGENIC FORMS According to this report. both tAl and 2A1 reference eigarettes contain substrates for hepatic mrMxwsygenases capable of generating metabolic inter- medutes that are muu6eme /or S r2oAimrnam tester strains TAIS711 and 7 Aak the laure n select:vtty more unsibve to muu6enesis induced by smoke I condensate. The most muwagenic franions, nmrcover, are not the o.es that contain polrcrclic aromatic hydrocarbons (PAN) but Ihose supposeMy con- taining such base-soluble chemicals as aromatic amiees. The data supest, therefore, that most (about S/9i ) of the total emtagenie activity of theae co.- dcnsates lies in the briic fractions and not in the ones containing the PAH. In addition, reouse puimonary tissues seem capable of activating certsi. PAN and aromatic amines but not others. Mouse, rat and husnan pulrnonary liswrcs are currently being compared for Iheir ability to metabolically activate cigarette smoke condensate euKrial to biologically active forats. Kouri, R. E. tr .f. (lrfkroMolosir.f Atsocl.tet. Inc.) and /enelkl, IY. F. In: Water, M. D. r1 d. (eds.): AppIkwrions o/ SAorr-Ter.w Alo.r..ys M rAe Fr.rrlonaion o/ Compk. Endronmenrd M/trwti, New York: Pkawm Puk« Inhin6 Corp. 1979, pp. 497-512. From the Department of Biochemical Onoeolo6y, Microbiological Arociates, Ine., lletheda, Md.; and Childreri s Hospital of Los An6eks, Loa Angeles. ORGAN SPECIFICITY OF INDUCTION OF ACTIVATING AND INACT IVATING ENZYMES BY CIGARETTE SMOKE AND CIGARETTE SMOKE CONDENSATE • The effects of cigarette smoke and eigarette smoke condensate oo the activating and inactivating erxyme syalerws of the body were atudied ktn by in vivo and in vitro methods. In the rat, ciprene snake ieh.lation selaedisely induced lung and kidney aryl hydrocarbon hpdroaylase (ANH) activily. Oa the other hand, eposide hrdratase (EN) and Rlwathione S-traruferue wat trot si6niAcanty modi/led in any tissues o/ thtt keated animala. Coropwed to the kidney AIIII, the lung hydroaplase was three-fow times wtore sensitive to ammaR concentrations of cigarette stnoke and aeemel Io Aave a longer k~ioloRkal k,.lf• li/e. Is bolh tissun, the induced ANN presented the sanse /n virn se.ailiviiy to various inhikNors as a polycyclic hydrocarboo-itrdreed ANN. 1s the /n Wn+ uudies, cilareue ar.oke condensate fractions (CSCF) induced both ANN a.d EH activity in primary fetal rat liver ceM erltures. NeverthekesR tAe ANN activity responded faster and so lower concentrations of CSCF thas did die EII activity. A{ow conceMrMion of bewt(a)antlwat:ewe imWced only the AHII activity, while rrrnr-stilhene oitde enhanced selectively the EN activity. Appropriate concentrations of CSCF or of phenob.rbital determined a paraRel induction of bowh enrymes. 11 seems, therefore, that the liver cell cuNwe eonstitwes a unique tool for a comparatiw study of the A111/ ud Ell In- duction mechanisnss. The eaiVence of coordinated or independent biochemical control of AIIl/ and EN activity is discussed in this paper. Gielin. 1. t. et. d. ArcAiv.r of To.kolorT. Suppl. 2(Mechanism of To.ic Action on Sane Ta{et Organs). 2)9-251. 1979. Other arrporl: Fonds National de Is Recherche Scientifitiue Mldkda. From the l.aburatoire de ('himic Midicak, Instilul de Palholotie, Lille, Belgium. 1 16 17

1 i A NI;W FIIGi11.Y SENSIIIVE ASSAY FOR ETHOXYCOUMARIN 1)HI:I/IYLASF IN CUI.IURED IIE.PATOCYTES Etho.ycoumarin, an imeresting and useful substrate, is deethylated by diflerent cytochrome P-ISO species. Although hepatocyles in culture are presemly considered as an interesting model for pharmacological and loai- cnlogical invesliAatrons, the study of their eytoehrome P-130-linked moraosy- genases reqrires snphiuicated methodologies to detect the usually very low enzymatic activities In thrs repon, a new, rapid, aensitive, and specifk technipue /or the eaasuremem of 7tthosycounwin deelhylase is described. This assay is based on the multrpk-ion deteNiow of the trireethylsilyl ether of 7-hydrosy- coumarin The new mNAod, developed for the study of cytochrnme P-I30- dependem nanooayRenases in cultured (etal rat liver eeMs, allows thr deleetion of activities as low as 0 2 pmol/min. It seema possibk that such a sensitive method may prove very helpful in the s1udT of cytochrome P-4S0 rnuluphcuy in small sampks such as cell culture and biopsies or in low-activity tisaues like the skin and lung. De Uraeve, I., K,emen, P. Frankine(, C., and Gkfen, 1. Anefyrit.f AiorArnd»r7 104:119421, 1950. Other .rpport: Fonds National de la Recherche ScientiflQue Mddicak. From the I aboratoire de Chimi. Mtdicak, Institut de Patholotie, Li2ge, Belgium APPARENT ABSF.N('F. OF RFQItIRFMPNT OF IIYDROCARSON METABOLISM FOR INDUCI/ON AN[) REPRESSION OF MIXI?D- FUNC f ION OX IDASFS TAis report presenla the initial results of a new series of investigations ' Into whether aryl hydrocarbow hydrosylase (AHH) induction and dimethylni- trosamine (DMN)-demNhylaae repression require priw metabolism of hydra earbons. Methyl substituted polywckar hydrocarbons whose subslNuents aterically hinder the metabolically important b.y- and K-reRion, as weR as derivatives with an epoay =roup N 1he same repions, were the invesliptive tools Iw thia study. The dlects of injectiwg these compounds into male rats 24 hours be/ore sacriAce were determined by measuring the DMN-demethylas. I and AIIH activities in the microaomal protein of the hepatic poumitochron- drial (raction. According to the data, /he enzyme inducing and repressing ae- ti.i1y of hydrocarbons is IndependeaN of priow metabolism (at kast at the bay- and K-reRion). These resulta con/lrm previous observations by others that various oa1eenated derivativn of )-methykholanthrene and bento(./pyrene are Cnnsistemly less potent than Ihe parent compound in their capacity to induce Alll/ in cell culture. The present data also support other unpublished reports on the aMnpclitwrn of various potyeyclic hydrocarbons wilh tritialed 2,1,7,R- trtrachlorodihen.o p dw+.rn for specdk binding to the eylosohc receptor /or AIIII rnducrrw. Arrrr M I. MYrir \( a.nJ .I'em. 1C 1tl I ChfrnJCo-8iofolkd fnterortion, 29:247-253. 1950. Other aupporf: IloQmana-La Roche, Inc. From the Scamen's Memorial Research Laboratory. U. S. Public Ilealth Sarvke Ilospital; and the Depariment of Medicine, Tulane University Medical CerMer, New Orkans. INTERACTI()N OF THE TOSACCO-SPECIFIC NITROSAMINES, MI:TIIYI.ETIIYI.NITROSAMINE AND N-NITROSONORNICOTINE, WIlll DNA ANI) GUANOSINE The in vitro microsomelalalyted ieteraetions of rrC-mnhykMylwitros- amine (MEN) and rrC-ni(rosarornicoline (NNN) with eaoRenow DNA aaed Ruano.ine were e.amirarl in Ihe hope of gaining an iwsiRht into the teecsa.ias of the carcinogenic action of Iobacco-speciAe witroaamines. The resuNs show that rat liver microsomes catalyze the covalent binding of the r'C-labekd aeg- ment of MEN to call thymus DNA, and suggest that the activating ewtyrnea are miaed lunction o.idnes. However. NNN binds only to Ruanosine and ael to DNA. The addition of cylosol lo the iwcubalion system markedly reAuee. MEN binding because demethyluion is enhanced more than deethytatio., a.d this in turn decreases the formation of elhykarbvniwe ion. That pretreNwent of the rats with phewobarbilal (PS) or with )-methykholanthrene deaeaaea MEN binding also supports the concept thal ethykarbonium ion is the reactive intermediate that binds to DNA and (hu demethylation is the rate-limitiq atep in its formation. Addition of bentylamise, a highly potent inhibilnr of DMN-demethylase, eornpklely abolishes Me MEN binding catalyzed by mioro- somes. Mitochondria do not have any sipiRcaM effect on MEN binding. Lai, D. Y., Arcor, l. C. and Argua, M. F. Re,r.rrh Comrnunic.rioru /w CherNkd P.rAdoty ir,d PharnarobRy 21(1) :17- 10), /980. Other arpport: HoRmana-La Roche, Inc. From she Seamen's Memorial Research Laboratory, U. S. Public HeaMb Ser.ioe I/osprtal; and Department of Medicine, Tub.e Usiversity Medical Center. New Urkans. . FACTORS INFLUENCING THE MICROSOME- AND MITOCHONDRIA- CATALY2tD IN VITRO BINDING OF DIETHYI-NITROSAMINE AND N-NI I ROS()PIPERIDINE TO DI:OXYRISONUZI.EIC ACID Although it is generally accepted that both the dialkyl and cyclic .i/roa samines rcQuire metabolic activalion by microsornal edaed-lunclion oaidases lo display their carcinogenic or mulrgenK acli.ilies, the chemical nature of a.y 19

I i presumed alkylating inlermediates remains unckar as of now. In this study. In rirro syslems were used to investigale the binding of /"('{diethylni- trosamine (I"CIDEN) and /'tCIN-nilrosopiperidine (1r'('INPiP), a higher dialkyl and cyclic nitrosamine, respedively, to eaogenous DNA. Results showed that both compounds bind covakntly to calf thymus DNA in an !n rlr.o in- cubation system comaining rat liver microaomn. This binding reaction is dependent on NADPII; a requirement Ihal is eonsistent with the involvement of a microwmal mised-(unclion osidast srskm. Prelrealment of the animals with phenbbarbita/ ( PB) enhances 1hs bitdity of both DEN and NPiP 1o DNA, whereas the binding of DEN to DNA dareasd aftet )-methykhotanthrene prdreatment. 7Le PB eRect, as observa+d /rom the binding of DEN to DNA, is more pronounced in young rats than in older ones. Addition of cytosol to the incubation aystem enhances the bJwdi.g o1 DEN three- 1o tour-fold and that of NPiP two- to tMee-fold. Addilion ef toitochondria to the incubation system increases the bindiwg of ("('/DEN only sNgMly, but increases tAe bind- ing of NPiP more than Ave-told. AddNion of tnNochowdria has tn effect on the binding of ('K'/ dinxthylnitroumine (/"CIDMN). Mwochottdria alone marked- ly catafyre the binding of NPtP to DNA. Additiotn of beazylamine, which is a substrate of mitochondrial monwmina oaidase u weM as an inhibitor of DMN den.ethylase, inhiMts the binding of NPiP catalyzed by microoomes and micro.on.rs plus aritochowdria. Lai, D. Y., Arror. 1. C. and Argu.. M. P. /ixArnslcd rr.nt.rolop 28:33e3-7330, 1979. O/her wPp.rsr Nofiman. ta Roche, Inc. From the Seatnen's Memorial Research laboratory, U. S. Public Health Service Ilospilal, New Orkans, and Departmeal of Mediciwe, Tulane U.ivenity Med- ical Ce.ter, New Orkass. INDUCTION OF DIMETHYLNITROSAMINE-DEMETHYLASE BY POLAR SOLVENTS Acetone snd isoptopanol both enhance dimethylnilro.amine (DMN)- demethylase activity /n rlro, and /w vitro reaetiows supest that these polar compounds do od acl directly ow the sniaed-function osidases. The /w v/ro enhancaneM of DMNdetmthylase I by scdone and itopropanol, therefore, may not be unique (o these p.rtit:ula oosnpovnds, but may in f.c1 he a prop- er/y of a number of slrucluraRy unrelakd polar mokeuks. Data derived from the in'raperitoneal administration of various pdar sdvents to rats confirm that acetone enhattcn DMNdemethylase I activity In .Jro, an effect also produced by dio.ane, tdrahydro(ursn, dimethyhulfo.ide, and trioctanoin. Dioaane elicited the highest increase is activity. and was also the only sub- stance that s.gn.lkanmly, if drgM1y, increased aryl hydtocatbon hyJro.ylate acU..ty /n ..e.a tMtc coenpourrl& rnh.Mt I/MN Jemcthylate I actuvrty. The actual n..dc.ul.. nw.A.w.aa p.wern.ng ,he represrwn and attwation uf mrera u.mal m.seJ /una..r .•..davs by tt+U.n themn.ta n unknown, but it hat 201 been proposed ahat these agents modify the conformation of regulatory pro- Ieim, thereby causing repression and derepression at the gene level. Polar solvenu are known to be poteM protein denaturing a"s at hio concew- tratioes but actually have a concentrationdependenl ambivaknt effect ow pro- tein conformation: they rabilize and incretae the heli>t content at low eo.- untrations and unfold it at high conceMrations. Recent and as yet unpub- lished results tend to confirm this conformaliow-aadifyinB effect since induc- tion and repression by polycyclic hydrocarbons do not require their maabd- ism. 'The data presented here ruk oul a" in ritro adivalit.R effed 'of polar compounds at the e.istind enzyme level. They ue no1 iuco.aistent, howevtr, with the possibility that the effect of these agewts-operaKt>t at Ihe level o/ repressor porsein(s) of newetic regulatiow of DMN-tuelhylas*-tnay play a role in the !n vho e.h..cemesn of erwyme activity. Argus, k1. F., Neuburger, t. 1., Myers. S. C., and Arroi, !. C. rrorrrd/np o/ the SorArtr /or Frrrrin.rntaf Riefop and Mrlkinr 16):32-33, 1980. From the Seamee'a Memorial Research I.aboratory, U. S. Public Ileahh Service Ilospital; and the Depattmenl of Mnlicine, Tulane Univenity Medical CettNS, New Orleans. I ROLE OF DIMETHYLNITROSAMINE-DEMETNYLASE IN THE METABOLIC ACTIVATION OF DIMETNYLNITROSAMINE Miued functioe oaidase modiBen-3-wtelhykholasithrewe (MC). /mg- nrnokorlbsarbonirrtrik (PCN) or a-ss.pMhoAavate (A-NF)-adtwiltia- tered to rats iw vlvo inhibit the /n vitro binding of dimethyfnitrournite (DMN) to DNA catalyzed by 1he hepatic miaosonses and NADPN. This efted p.ralkls the one on mkrosornal DMNdeme(hylase 1, eonsidered the only .c- tivatinR enzyme system in DMN carci.ojewesfs. However, MC ..d PCN inhibit the hepatocarcinWnic activity of DMN whik $-NF enhaeces it. la mice, a-NF faso enhances DMN-demethylase 1, but does swt affect aha ep/a chrome P-130 level. On the other hand. PCN inhibils (he erzynM, kready elevating Ihe P-I30 kveL Neither twodi/kr alten the LDs,. la a dosasespos. study of hou-mediated DMN mulapnkity as Nfeded by these erodiAers` IM only notable effect was found wilh /he Aighesl DMN dose (200 rne/kR) when PCN pretreatment signi/kaNly bNesni/kd /he tnutaBenk rrespo.se. AI- ternates for the established DMN metabolic pathways were considered in order to clarify these anorndous observa/ion.. Nuclei had previously bee. thought lo be involved Iw DMN activation, but nehher whole rat Nver wucki twr isolated membrane fractions showed any DMNdewtelhyl.se or dic+1* nilrosamine-deethylase anivity, as re8ected by formaldehyde formation. Prr- thermore. neither purified amine osidase nor whok mkro.omes teltasd formaldehyde or oaidized NADPII in the presence of DMN, indieatiwg that Ihe /atuer is not a saburate for hepatie mKrouwnal .mine osidase. Though benzylamine. a mitochondrtal mono.mine osidase (MMO) inhibilor, was ahe 21

found lo he a potent 1)MN dcmAhytase inhibilor, the data do nW suppnrl the n.rtron that mKrnlMral MM(1 nKtabotltes 1)MN Thus. the cunllKting eRects n/ A NI: on 1)MN mcuh.drsm and carcrnogenesn are probably 6nwmrkd rn cetlnlar events unrelaled tn nArosamine metahrdrsns arwl the endrqdasmic rehculum, such as the shnnrlation o/ cyloplasmre sransport and/or DNA repair by the rnodr/ier. I ai, O Y. Myers. S(' , Wno, Y-T., Greene. E. 1., Friedman, M. A.. Argus. M. 1; , and Arrot. / C. CAn.rko enrloRir.l Innrurrionr ±ol:107-12i, 1979. Otti.r ...pperf: IloRmann-I.a Roche, Inc. From the Scamen's Memorial Research Laboratory. 11. S. PuMic I/ealth Service /luspdal, and the 1)epannsenl of Medicine. T+dane llniversny Medical Center, New (>.kans; and Binahemaal Tosicology, Corporate lechrw+h.gy, Allied ('hemrcal 0wp..ratwn, Morrrslown, N. 1. ENIIAN( 1 MENT OF IOXI('ITY AND FNZYME-RI:PRF.SSIN(: A('TIVIIY OF l-UIUXANF BY ('N1.ORINATION: SIFREOSFLE('l IVE 1:FFP('IS In order to elucidate the structure-aclivily relationships and mechanism of action of din.ane-type cyclic ether eareinogcns. Ave chlorinated deriw- tives of dinaane wcre synthesized and their losicily and ability to metabolize drug nsetabolitrng entymes were investigaled. AN the chbrinatcd compounds were /oJnd to be at least 10 umes more /o.ic than the parent eompound, Ihe LD„ of which was previously delermined to be S.) g/tg. llse 2r, 31. St. 6c-ICtrachkxo derivative was.the mosl loaie compond with an of 5.3 mg/tg, corresponding to a 1O(11) fold enhancement of the loaicity. Steric con- figuration obviously plays a majnr role in determining the tosK i/y af the derivatives lhe ?r, 11, SI, hc-isorsser (isonser 11 of Ihe telrachloro derivative was Iltl Iinsea more losic than its 2r, k, St. 61-eouMerparl (isonset 1I). The Islter compound was dw a potent repre.sor of hepatic dimethylnNrosamine- densethylasc 1(DMN-d) and aryl hydrocarbon hydrosylase (Allll). From a practical point of view, this study denwtsslrates a dramatic enhancement of acute Ioticlly of p-dioaane by chlorination. Whether this is accompanied by a similar change in the carcirw+olenicity is curtently under study. Wro. Y-T , Neuhurger, B. 1, Arros, 1. C., Argus. M. F., Nishiyanu, K., and Qriflin, U. W. loairoloty t.rNea 5:39-73, 19140. Otli.r support: National ('ancer Institute. Cancer Assoeiation of (:realer New Orksns, and I/o(Imann l.a Roche. Inc. Fr.nn Seanren's Memrwial Research I aluqstory. If. S Puhlic Heahh Service 1(ospitat, New Orkans; lkpartment of Medicirse. Iudane University Medical ('enter. Ncw Orlcans, and the 1)epartmenl of Chemrslry. University of New (Llc.ns R1:1)ll(-11(/N 01: NI('UI INAMIDE ADI:NINE 1)INU('1.1:.O1/1)li I.EVEIS 11Y UI I IMA 1 F('AN('IN(k il'NS IN IIUMAN I.YMPII(K'Y 1 E-S In Ihi. search for a pmsihk correlation hctween DNA damage and lower- ing rrl nicutmamide adenine Jimrckolsde (NADI kvel.. the eflecl of several classes of 11NA-damaging chemicals was studied in freshly isolated huwsan lymphocytes. (N the 21 compounds eaamined, seven were direct DNA-dam- agrng agents and 14 were either non-DNA-damapnot compounds or metabolic activity was rewsireJ to cause DNA damage. Rapid lowering of cellular HAD kvels was caused by each of the direct DNA-Jam.llinll chcmicals-enamiaed including N-methyl-N'-nilro-N-ailrosogssanidine, melhyl methane wl/onak. N- acetu.y-3-acety)amim+flrwwene, 7-hrumornelhyl6en:(a)amhracene. and Ihe bea- so(«Ipyrcnc derivalives, r-7,r-N.Jdsydroty-9,10-epoay-7,R,9,10-IClrahydrohenro- (w)pyrene and hento(.)pyrene-1,3<poaide. lhe issdirect-scling carcinoolen 2-acelylaminnflunrene, 7) polycyclic aromatic hydroeathons, and derivalires that were mm•1)NA-damaoling did nol eawe lowering of NAD. These resuhs provide strong evidence for a telauunship belween DNA dsauge and aaMe depression of cellular HAD pools. Rsnkin. P.W., lacobson, M. K., Mitchell. V. R., and Aribrr, b. L. C.nrrr RrsearrA 10(6):1B0)-111(17, 1990. OOther swppv.rts National Institutes of HeaMh, American Cancer Sociely, aa/ the North lesas State University Faculty Research Fund. From the Deparlmenl of Biological Scienees and Ihe Genetics Center, Nott` Tesas State Univenily, DeMon, and Ihe Departwsents of Chemistry and Bio- chemistry, North 'Tesss State Univenily/Tesas College of Osteopathic Med'a cine, Detslon. ('ORRFI.A f1ON OF CARCINOGEN-INDUCED UNSCHEDULED DNA SYN11I1:S1S AND NAD REDUCTION IN FRESH HUMAN LYMPIIO('YTFS Incorporation of aH-Ihymidine Into the DNA of fresh human lysnpho- eytes Irealed with various chemical sswtatens was nseasured and eorre/NCd with cellular NAI) levels before and afler lreslmem. The HAD IeveM M lymphocytes were significantly redsteed following IrealmetN with mulaolenk chemicals. Reductwn of cellular NAD pnoh was directly correlated with sH thynsdine incorporation. As NAI) kveh decreased. s11-Ihymid'uw iacar- pnraliun increased. 1)senplsyllin< a)tnown inhibilor of po/y(ADP-riboa) - prAynserase. inhrhite.l both Ihe NAI) redueliors in eells treated wilA DNA danlaglng agents and 1he inc.upowlion of '11-Ihymidine in/o DNA. The inhibitory effect of Iheophyllinc on NAj) depietion and on aH-thynsidiae uscorpwatinn was dose and cell mrnsher depenJenl. Nesr rsormat reopewse /o carcrm+gen e.prnore could be restnred to Iheophylline-Ires/ed cells follo.- Ing she rensoval of theophyllirse I hesc Ja1a suggest that eonversion of NAD to poly(AI)P rth.se/ may he necessary to. or al least closely associa/ed wilh, DNA repair in human lymphocyles. ?1

Brtbr., n L. .r ul Cenr.r L.rtns 11 1 S) 1611, 1960 Other au'p.ri1: Amcncan Cancer Society. National Institutes of llealrh, and the North /caas State llmversny Faculty Research Fund. From the Ikpartnments of Biological Sciences, Pharmacology, Chemrslry, and Bruchemnlry, and the (;cnelics Cenltr, North Tesas Stale Universny, I)enloe. SH UNII/M FFFF('TS ON 11/E CARCIN(XiENIC1IY AND MF.1 AM()LINM ()1: 2•A('li I YI.AMINOFLUORENE Selenium has heen showw to reduce the rate o/ chemically induced tum.w in esp:rrmcn/al ammals /n viro. In Iha search for the hasn rd the n.wed e8e.1. MMh rn rirn and in rpre testa were carried out and carefully evalualcd In the in rir.r slydics, adddion of (our ppn Se (as Na,Sen,) to the drrnking w.ner of mak albino r.N fed diets containing 1101% 2-ace. tylamm../tuorene (AAF1 povrded protection against hepatic damage and rrsuhed rn at least a e10% rtduclinw in liver lunsor incidence. An in rirro assay system utilizing mKrotomes from Na,SeO, supplemented or nonsuppk- mented 3 merhykhulamhrene induced rats was used to determine the effect of nral Sc mtake on the metalwdism of AAF. Oral Na,SeO, adnunnlra/run led to an increase in ring hydrosylatron and a decrease rn N hydro.ylati..n. AdJnw.n of Sc to the mrcroaxrral assay system increased l-/l/l AAF /orma- tiun and decreaseJ NO11 Aff /ormatiors: lhe observed inhrMtion or slowing of AAF carcinogeneus may he the resull of the decreased poJrrctinn of Ihe highly carcinogenic and mut.genie metaholile, N U11 AAF, and the increased production of the noncarcinogenic rrng hydroaylaied me/aholnes of AAF. lhese data indicate that Se may cause a shifl in the metabolism of AAF toward deluaifkation. Marshall. M V., A.nwr, M. S.. /acohs, M. M., and Grdlin, A. C. C.r,ri. /..ners 7. l l 1•) lll, 1979. Other .arprlr T. K. Dison, The Robert A. Wekh Foundation and Bio- medieal Research Support Grant. From the Ikpartmenls of Biochemistry and Biology. The University of Tesas System Canctr ('enttr, M 1). Andetson Hospital and Tumor Insutute, llous- lon; and The Eppley Imlitule for Research in ('ancer, Omaha. EPAXIt)1; IIYf1RATASE lhis eslensive review summarizes al kngth the latest adv.m:a relalinR to the true role of epside hydrat..e. Numerous observations ukcw together suggest that a single enzyme in microsomtal memhranes hydraws styrene o.ide and henzo(„)pyrcne 4.5 oside, and that this enzyme is eposide hydra- /ase In its pre form, thn has a minimum molecular weighl of 49,000 and a hiRh rendcncy to form very high m.)fecular wtighl a"reples, wfirch agrees I with its hydr..plw.hic character. (1 al.u has a broad specificity far substrate epo.idc., inchNlmg tho.e .krived Irom carcinogenic polycyclic hydrocarbons. Ihe cnzyme, /urlhcrnwrre, appcars 149 he uhryuitous in the urgans and species invr.liCaled, suggesting N plays a vital tak. Its high activity level in kaes aml uvaries irKlkatcs either a nde in skraid melahulism tK a proleclive role lor the gunads Based on invcsli6ation. to date. it appears qualitatively simiar in Jr(Ierenl satans of the same species ahhrwgh specific activities can differ by as much as three orders rq magnitude. 1/ also appears to he qualitatively simdar in drffcrenl slrains of the same species or in diAerenl phylu=cnelicaRy cluscly relatcd spxies, fnrl rwN in phyh>tenetically nwre remote species. lhere- fore. the el+rsaide hydralase found in these phylogenetically diAerenl groups musl he functionally different forms. h is still nM known. however, whether these /unclwnal di/lerences are the re.rdts o( ddfereM enzyme prouins or of dJlcrenl memMane environmenls. ()erch, F. In: Bridses, 1. W. and ('has.earNl. 1.. F. (eds. )• r.aKrca. In Drug MrrnAolisnr. ('hichestcr, England: John Wiky, 1979. Vrr1. ), ehapt. S, pp. 23l-lol. Front thc Ikpartmenl of Pharmacolosy. University of Mainz, Mainz, West (hrnmany. IN I FRINDIVInUAI VARIAl1ONS OF EPOXIDE HYDRATASE ACI IVI l Y IN HUMAN LIVkR AN[) LUN(i BIUPSIES, LYMPNOCYTES ANI) FIBROBLAST ('UI:fUR1:S Ilunsan epuside hydralase tFiil activity was measured in this study is biopsy samples from the liver (which is 1he main site of drug wrclabolitr~) and Irons the hrnr (an organ which ia coMinuaRy ettposcd 1o potential ess:ywm inducers as well a. 1o carcaw+genie companads). '1he !:N was dso studied in native lymphocytes and in cuhu.ed lyrwplwcytes and RMoblasls. Results ahowed that the sp~.tifk activity of 1?l/ varied 17-/oW in liver mkrosorras Iruns 71 d.Nwrs, and thal there was great varialion iw the other mtasurtnseMs. Spccdically the .tudy shrwed that 1:11 activity varies considerably /rons organ to organ and, in wnss rwRan.. /asns individwl to Individual. This was par- trcatarly apparent in the livcr and hmR. organs heavily esposnl to fwtipt c.unpwmds. 7 hcrc was much kss variatian in cuntrol liver and in ee1H in crdtaue under unilmm environmenlal eanditions, indicaling that epieenetk lacl.ws are esenlral /rx the differences. Several paramelers were found to currrlalt with allct.yl acrivily kvel.. In the lunww itself EH activity was ckarly radrkcJ. A nurkt.l mcrca.t in liver III activity was oMerved in pa- trcrNS IrcatcJ Nith ..rnsc drults ,A11MMrph variation In F/1 activity may mN hr tau.cJ E+en.tircally. Jdterrnccs d.r esi.1 and i1 ia very likely Iha1 17-fold Jdlcrcnres. as f.wusJ in Ihe hscr in thr..tudy, have impurtant hw>w.llical in• pIKa1N.n. cnhcr in the metatrJr.m 411 cral..t;cmwc compuunds or in response to IlrrC11',n l'.MlpNmds. (:latu, 11 K.. I..rrnr. 1. I ki.alnu.rmr, R., Rcmmrt, 11., (/hnh.nn, F. F„ Kal- Ienhx'h. 1: ,/rgtnrcycr, 1. Rrhhrcr. 1/ . arw/1)rwh. F. In: ('otnt, M/.. Cumrcy. A. 11 . 1.1aM.w.k. R. W.. Grlhoin, 11 V.. Gillette. 24 ?S

1. R, and O'Sricn. P. 1. (eds ): Alk.o.onwr, Drrr O.idaaioru. and Chrrnkai C.rcinoerwrnr. New York: Academic Preu. 1990. vol. 11, pp• 6S1-6S4. From the Phy.nalologischcs Inslitut der Univenitil, Mainz. West C.ermany; UnivcrsitYtskbnit und foaitologische lnaitul, TUbingen, West Ger nany• In- selspital, Sera, Sww:erland; and the UniversilGskrankenhau. Eppendxf, Ham- bwt, West Germany. TRANSPLACENTAL TF.RATOGENRC AND CARCINOGENIC EFFECTS IN RASSITS CIIRONK'ALLY TREATED WITH N-ETIIYL-N- NITROSOUREA The rabbit wiwh its hen.ochorial type of pl.cems. sintilar to tiul of hu- rnans, is an eacellent compsrari.e mode/ to inveuiga/e Soth lrsnsptacenta/ earcino4e.esis and IerMownesi.. UlilizinR lhia convenient wadel, Iherefore, the ef/ects of low-level chronic administrtliow of N<thyl-N-nitroaourca (ENU) have been dekrnwned in the rabbit. For this study, pregnant rahbits of two putisMy inMed slraina. WH/1 and IIIVO//, wrre given lew consecutive dail7 i p. iniecKorn of 10 mg F.NU/k@ dissolved in /rioctanoiw starting on either day 1~ 15, or 11 of gesution. Of seven WH/ 1 progeny weaned, three devel- oped primatity, renal lumors a1 6.1=0.6 months of age. Similarly, of 12 IIIVO// prowny weaned. 10 developed primary renal tvmors at 6.)t0.6 months of age. In addd'an, one AhroMntic oaeosareoms and Ave wenro- Ahrornas, sorne associated with wewikrnms eysb, were observed. The (re- "ney of tumor induction fot each strain was similar to that observed in previous espetiments with acute administration of ENU andethylurea plw N.NO„ k.rt the latency period was aMnost doubled. In addition, tetatogewie eAec1s were seen at birth. In one eAect, a disproportionate stuMing or minia htrization wr evident, the frequency oe which inereased with the age of the treated fetuses; in a second eAect, holes occurred in the parictal sones. No leratownk el/ects were seen in the controls. Foa, R. R., Mrkr, H., Potlathil, R., and sedi=iaw, H. G. Journal o/ rAe National C.nrrr 1n.Ntrre 63 (3):607-614, 1980. ()1Aer sr'r.r/: National Cancer lnslilele and the National Inslitutes of IIeaMh. From The lackson laboratory, Rar Harhor, Mc. RETINOID INIIISITION F.XPERIMENTAI. LIN(3UAL CARCINOGENFSIS: ULTRASi-RUCTURAL OI/SERVATIONS It has bees shown that vitamin A and some of its synthetic analogs can inhibit carcinownesis in a number of eaper(mental awtwr systems, but the mechanism governing such inhrbitiow h not well understood Since initial nudin h.ve demonstrated the inhibitory effect of II-rlr•retieoic acid on ear- cimrwncvs of the oral mucous memhrane, however, ultrntnxtural studies seenrcd profltabk ('onatiuently, four groups of male and female Syrian golden hamsten, a total of 64 anirnals, were etamined: in group I the posterior lateral border of the animaM' tongues ..ere pai.led with a 0.3% sohNioft of 7,12-dimethyl-ben:/.)anthncenc (DMSA) in .celone• group 2 received Iir same treatment plus 10 mT of orally administered I3-els-retinoic acid is pcaaM oil twice weekly; group 3 was givew the retinoid only; roup 4 consisted of untreated controls. The aeirnah were the. sacriReed N regular idervals dter the Ara It weeks. Initial resrlls of uhretrncwral Mudies (eksetroe r.iua scopy) wqer that the urwors which developed in the retisoid4reated aai- mals were better di/ferewliated thsw those indrrced irt Nr awirt.als not nce(v- in6 the retinoid (group 1). Vitamin A is known to poornwe epilhelial ceM ddferenliation, an effect that tnsy block the dediQerentiNing action of DMBA lhus accounting for the obaervations reported here. ShHr, a. rr r. Iorrwel o/ the Nariond C«aer Inrrqure 63 (6 ):1307-1317, 1990. From the Department of Oral Medicine and Oral PahokM, and 1_aboratory of Electron Microswpy, Harvatd School of Dental Medicine, doaton. I GENETICS IN UROGENITAL CANCER: HEREDITARY VERSUS ENVIRONMENTAL FACTORS Certain cancers of the rroAeatal syslern have beert ascribed /o pri.wy Aenetic facton. although a variety of studies have also ahowa that envire.- ntental iwteraction may play a cotrribwory role. l0 this paper, sections an devoted to the following wo6edtal ea.een srhere{w primary Aetretic faturs aro considered to be etiokVic: trawsNiood oeN caeiwowta, renal «N ear- cinana, von Hippel-LMdw's disease, Wilws s Inntor, .cphroblastotw. a.eW eiated with disorden of posrqr awd oorrAe.Nal t>wdfonnationa, eawoen af Ha prostNe, Iesticular cancer, and fnnili.l Ieralor of Nre 1eMes. The noosds considered in Ihia study did, indeed, shew IhN Aenelie faeton appeared te be operative in a signi/karrt wrwtber of pa1k.N, producing InAtMed predk+s positiows 1o the development of Aertilowiwry eatoen. 'flrcse hi61t-rak patksas could anticipate their ttlmors to occur al yottwAer aAes awd to be rwuNifocal or bilateral in paired organs. They eould eapee tlye recunence of new uv similar kt.ions with the passage of lime, despils earlier appareet ewdive /retl- rnent. F.arly and contiwinR periodic swveiManoe with appropriate diaposlie lechnipues throughout their lifetime seems to be essential to a successful ou1- eorne. /lowever, genetic predispothioM was oltew adrNi.ed with adverse ewdr- onmental eeposure 1o carcinogenic (aclors, making deAniliow of cancer etblosy difficult. A failure to appreeiate Ihe hered'Mary aspects of wolienitsl cancer has delayed medical knowledge and prevenled obvious therapeutic and control advances in this area. L.ynrh, H. T. and Wal:ak, M. P. drolook Clinks o/ No.rh Amrdra 7( l): e I S-e29, 1980. FFrom the Ikpartment of Preventive Medicine/Public HeaHh, Creighlo. Ual- veney School of Medicine. Omaha. 26 27

(:AI A(-TOSY1. 1 RANSI-FRASF: ACTIVITY IN IIl/MAN l RANS.- llONA1. ('1?I 1. ('AR('IN(1MA LINES AND IN BI:NIGN AND NI'OPI.ASI IC 11UMAN BI.ADDhR EPI f IIEI.IUM Methods for the assay of {a/.ctosyl /rans/erase (GT). as well as the propertics of Gf in estahlnhed human Iransitiond cell carcinonu (ICC) Irnes, are descr/bed here lhe (:T assay, which was run on cullurrd cells of Auman'f('C line MGII UI in wipewsion, nseasured the lramlcr of ('11) galac- tu.e from uridine diphosphate: (sHI galactose to dcsialylatcd ovine submas- dlary mucin. The a.say was optimized with respect to time and to protein, uridrne dlphosphale.galactose, desialylaled ovine suhmasillary mlKrn, and Triton X-1(if/ concenlt.twns. This asuy was then applied to fresh specimens of beniRn. infiamcd. and neoplaslie bladder episheMrm from )) palrents who underMenl cold cup biopsies al cystoacopy. Transitional cell earcinuma speei- mens gave values in the range of 24 7 to 184 • cpm 1'11) 6alaNr»e trans- ferred per pt protein per hour; normal and inAuned specimens ranged from 0!1 to 46 1 cpm pt protein per hour. By using a known method of cell tuplure, cell ghexls, representing cell surface membranes, were isolated both /rons the cultured cell line and /rom Iwo biopsy specimens of 1C!'. Though a complete enzymatic and elecuon rnicroscopic analysis was not undertaken, the coincidence of an enzyme marker with the cell Bhost (raclion arnlaining the elevated (iT nmade rt appear prohahk that Ihis enzyme is located in the cell sur/ace Sonme preliminary esperimenls were also tun to determine whether shed bladder cells in mrne wrwdd also ahow elevated (iT activity in cases of bladder cancer lhe resulrs of Ihese IeHs indicNe that this assay may prove useful in the development of dutnoslic fluoroimmune or radioimrnune leeh- niquea for dHecllnn of 1 C(- Plotkin, O. M. Wldes, R 1.(liR+en. S I- Wolf. l: . Ilaten, I K, and Prou1, O. R !r. Cancer Researrb )9: )S36 )569, 1979. Other wrpart: National Cancer Institute. Frorw the Department of Nutrition and Food Science, Massachusetts Institute of Technobp, ('ambridte; Urokogical Service. Massachusetts General /1os- pital; and DepartmesM of Surgery. Narvard Medical Schod, Boslon. VITAMIN A AND GALACTOSYI. TRANSFERASF. OF TRACIIp.AI. EPITNELIUM Galaclosyl Iransferase, an wnportanl enqme that mediates transfer of galactose to glycoprolein, has been described before for serum or for human bladder epithelium, but never for tracheal epithelium. In the present study. an assay was developed for this enzyme in microsomes of rat tracheal epi- thelium and the properties of this enzyme system were characterized. ResuNs nl feeding esprrin.entt dnne here showed that vitamin A defkrency greatly depre%sed t.laca.%tl Irsn.fela.c a.hvuy In vnvo, activity a.ukl he restored .oIhon e11 M.un by dr.unt Ihc anomal with rctUnyl acclale. Although adding I retinol to the microsomes frorn tracheal epithelium of vitamin Adefkienl rats also restored plactosyl Iramferase activity to srnne eslenl, full reslora- lion was achieved only by preincubaliog relinol with the microsomal prepar.- tion for 30 minutes. Preincubatiun with relinyl phosphate and retinoic acid stimulated Rdactosyl transferase activity in microsornes from /racheas of dl& cienl rats: preincubatioe with dolichyl phosphale, anhydroretinol a.d the ditnethylacetykrc{opentenyl analoll of retinoic acid did no1. These resulls iw- dicate that vitamin A is involved in /he plaaYo.yl transferae system of ra1 tracheal epithekum, possibly by being linked 1o placlose. Plotkin. G. M. and Wol/, G. Abr/ilmke rr itiopAyrk. Arr.61S:94-102, 1980. From the Ikpartmcnt of Nutrition and Food Science, Massachusetts Innituk of Technology. Cambridge. Maa. 11. The Respiratory System CLINICAL AND PNYSIOLOOICAL MANIFESTATIONS OF BRONCIIIOI.ITIS AND PNEUMONIA. OUTCOME OF RESPIRATORY SYNCTIAL VIRUS Epidemiologic evidence indicates that /he frequency and severity et p.l- monary symptoms in laler childhood and in nduMs are related to a hialorp of respiratory disorders during early childhood. Respiratory synclial srirw (RSV) is the mo.t itnpor/ant of the respiratory pelholienr in the Rrsl few years of Ii1e and the major cause of brrwrchiolilia aad pneumonia ln IMs ye Sroup. In this attempt W characlerue Ihe individual mosl likely b desKlop the acute and IonB-term complications of RSV infeclion, the phrsioloBk als. normalilies and clinical eorrelales were 1Mdicd in 32 consecutively hapilell- ized infants with kswer respiratory tract RSV disease. Oaimetry was wled M determine daily arterial osygen trturation (SaO,). Ap itifaMe wete lyrpat- emic on admission, with a 71-95!L (87'1k mean) S.O,. The lowest Iwer recorded dwwlR their hospNdi:Nion was 83.5% (3)-96% range). There was little improvement In hypoaensia during this period and nntil I 1o 7 weeks later. Significanl correlalinw was found between the severity of hyposetnia and the duration of viral sheddlng, otea.rence of apnea, respiralory rate, percentage of immature neulrophils in evidence, and age. The clinical aeverily of the disease dMd not currelate with the degree of hyposemia. Hdl. C. e., 11a1/. W. 1. and Specrs, D. M. Amrrk.n loarnaf of fflsesKS of (-billAoo! 1)):79{-002, 1979. Oth.r wrp.rtr National Institute of Allergy and Infectious Diseases. From the Departments of Pediatrics and Medicine. Univer.ity of Rochester School of Medkene, Ruehestcr, N. Y. ?y

RF•SPIRA fORY SYNCYTIAI VIRUS SF.ROI.OCiY BY A SIMP1.11:11:.1) I:.NZYME-LINKI?1) IMMUNUSORBFNI ASSAY Respiratory syncylial .irus (RSV) is the most impcxtanl respiratory pathogen ot infams and young children and the major cause of bwer respira- lory tract disease in 1ha1 age group. Recent evidence suggests that lower res- piratory tract disease in infancy may be related to the devetopnent of chronic or recurrent pulmonary dncase. In order to facilitate ideMiAcnion of RSV for epiderniobgic srudres, a simplilled eazyme-linked immunosorbenl asssy (I:1.ISA) wing commercially available reagenls for RSV-specific 1t(l has been developed. lhe EI.ISA method is desirable for epidemiologic srudies in that it is a reproducible and oAieetive teN, using small amounts of serum and afiordinA resulrs in days raher than weeks. This ELISA for I2SV ser- okly proved more sensitive thaa the standard complement Asation ur micro- neulralitatiogs tesrs in a group of 32 RSV infected adults, and there was cbse eorrelalion with complement Asatiow serological testing in 21 patients. lhe rse of purilkd amigens might allow she development of a more sensitive ELISA. SteinhoR, M. C., H.II. C. S and Schnatxl, K. C. lor.nd o/ Cflwk.l MkroBiology 12:447-450, 1960. Other a.".rtr National Institute of Allergy and Infectious Diseases From the Departmems of Pediatrics and Medicine. University of Itochcsler School of Medieine, Rochester, N Y. THE IMMEDIATE EFFECT ON LUN(3 FUNCTION OF SMOKINr3 FILTERED ANI) NONFII.TERED CIOARETTES ~ 'TA}s stud7 conArtns the obrsr.Mion that cigarette smokinA significantly Increases airway resistance. However, comp.risoe of the chances in pulmonary function induced by snwking Ahered and nonA/tered cigarettes with the same 1ar and nicotine composition showed Iha( AMen reduce some of the constiw- eMs that d1eN the large .nd central airvaln. Meawrement of instantaneow maairwal esphNory Aow rates (Vm.a„ ud Vmaa ,) and airway resistance (Raw) determined a significant increase in Raw wit~i both types of ciAaretles, .nd a smaR Iecrease In Vmas„ with the nonAhered cigarette. The change In Vmaar, obaetred after MnokinA the A1lered cigarette was no1 eonsidered sip- niAcs,wt. According to the previous evidence. Ahers decrease the irritant bron- ehoc•o.stridive eAect of tob.eco smoke, probably by reducing its content of tar a.d particvlale matter, ahhouAh a smaller amount of nicotine is also Irapped. Taveira ds Sdva, A M and H.nrorA, r. Awa.l..w lLdi.. of Rirrrarn.r f)be.ui 122(1) 794 797, 19110 I n.er rh. Itop.ftn.rwe ..t Wrdu.k Physiology and Biophysres. Georgelown tin..eosdv w.ah.ep„w Ir I PRI:VI=.NIION OF 111.ASI'ASE-INDUCED EXPF.RIMI:NTAI. EMPIIYSIiMA BY ORAI. AI)MINISTRATION 01: A SYNIHETIC ELASIASE INIIIBITUR Induction of pulmonary emphysema In laboratory animah by aeroo- solication or instillation of elastolylic proleases into the lung hs been widely used as an animal model of hunwr chronic obstructive pulmonary disease. 1. the e.perirnems presented here, a potent inhibitor of elaslase, methoaysucci.yl- alanyl-alanyl-prolyl-valine-cttloromethyl ketone, was administered lo mice oraNy to ue whether the proteinase inhibitor would prevent or dimiwih the scverN7 of elastase-induced esperimental emphysema. Resuhs showed that mice give. an intratrache1 dose of porcine pancreatic elsstase denrorutrated, after lorr weeks, an average meas linear (ntereept iw the hrwAs that was more than twice the normal value in untreated mioe. Other .rioe sLwNaweously lreated wish porcine pancreatic elaUse also received a siwAle dose of 300 to 400 H of methyosysuccinyl-alanyl-alanyl-pdyl-v.tineshbrosnethyl ketone (dissolved i. 0.3 ml of water) by stomach tube, al times ranging from two hns. before i.- stillatioa of the enzyme to 48 hrs. afler the enzyme. Mice thu/ received chlose- rnethyl ketone within 15 min. before irn1i11Miow of elslase were protected from development of emphysema, and their hmp were compktely free of alveolar deformation. By contrast, no signi/kant protectiow was aAo.ded by IttNwreN with chloromethyl ketone one hr. or more before challenge wish the ewyme or at any lirne after challenge. Chbrornethpl ketone done did rwt aAect the mea.linear intercept. Although many ques(ions remain concerning /he loaicity, imrmmoAeniciy, and carcinoAenicity of cMoranethyl ketones, this eaperirne.td model suggests that oral trealmeM with Ihesa aAeMs might prove useful in preventing pulmonary emphysema in persons with .,-aMitrypsin deficiency. J.woo. A. and Dearing, R. Arne.k.n Revkw of Rtr/L.ro.y Dluuse 121:1023-1029, 1950. Other ..r'.rt r U. S. Public Health 3erviee. From the Department of Pa<holoAy. State Univenity of New York at Stoq Brook. Stony erook. I PARTIAI. PURIFICATION AND CHARACTERIZATION OF MOUSE PERITONEAL EXUDATIVE MACROPHAOE EI.ASTASE . The part played by macropMAs elast.n In the reewdelinA of bry elastin in pulmonary ernphysesn. ia unckar. Research Ina aht p.rtidpriat of this enzyme in the phrsiolo~ic ~and pathologie turwover of e1s1iN world be helped enormously by the purilkation and charaeteriratiow of this enzyme. Previous attempu by others have used a eorwbinatiow of several ehromatoll- raphic procedures. lAe present reporl descri6ea a sinAle step dBrrily chrosna- toAraphy procedure for the puri/kaliop of elaNase from (he cvNwe mediww o1 mouse pcrironcal eaudative macrophyes. The etlecls of various nalwaMy occurring and synthetic inhihitors on Aus enzyme are also descrihed. AAi.i1y chromatography against SI)S Irealcd ,.-elaslin covalently linked to agsrosa ll w

, heads yiclded puritkd mouae periloneal e.udate macrophace clastase wilh 60% of us initial activity Based on Sf)S-acrylamide `el ekclrnphoresis, the enzyme has an apparent M, of 26,500 With molecular sieving chromatosraphy on Sephadea ael, macrnphaffe clastsse has an M, of 21.000 to 28.41114). It is inhibited by hDfA, but not by pancreatic and kukocyte elasla.cspecifk chhwomethyl Wone inactivalors or by phenylmethylsulfonyl fluoride. Thus, it appears to be a metslb p.otea.e. Macrophage elastue is also resistant to human .,•pro/einase inhihitor and to buth human and mouse w,-macroploMrlin, indi- caling, therefore, that i1 may have an important role in the physiologic and paholoeic remodeling of connective tirnus. White, R. R., Norby, D., Janoo, A., and Dearing. R. Sr«1lirnke rr flboAy.ka Arr.612:21)•211, 1990. Oth.r arrP.rft National Inslitulesof Nealth. _ Fraw the tkpartnKnl of Pathology. Slate University of New York at Stony Itttmk. Stony Brook. SF('RETION OF AI.PIIA•2•MA('ROOt.OBULIN BY IIUMAN ALVE(11.AR MA('ROPNAOFS Alpha-2-mactoollobulin (.2M). one of the major protease inhibitors found in serswn, is capable of inhibiting many bacterial and mammalian enJnpepli- dases. This paper reports the secretion of .2M by cultured human alveolar macrophages, thus indicating a possible local source of .IM in the alveolar spaces. The secretion could be blocked by 2.0 rg/ml eyclohesimide, an in- hibilor of protein synlhesia. Iktection of the Inhihitor was achieved by radio- imrmrnoeketrophoresis of concentrated euNure mediwn obtained from cells cuhe,cd in the presence of s'S-wmthionine. The sensitivity of the reka.e to low levels of eycloAesimide, and the /ncorporation of "S•melhionine into w2M wbonil proteins, support the contention that .2M Is synthesized and secreted by alveolar macrophages. The incotyoration of Iabel into protein with a moletulat weieM of 113 000, corresponding to the .2M suMmil, also suggests that the w2b1 recovered from the medium of cultured alveolar ntacrophates was itself hbekd, and was not simply eompksed to labeled prateates secreted by these eelh It seems, especially, that aeeretion of w2M by alve.dar macro- phages may facilitate the inlernalaation and disposal of proteolytic enzymes in the alveolar microenvironmen/ and Ihw may play an important rok in the defense of tAe lung. White, R., JonoQ. A. and Oodfrey, H. Lung 137:1•6, t91t0. Otibi support: National Near1, I ung and Blood Institute. Frnm the ()erartmenl of Patholngy. Slate University of New York at Stony Btonk- Sinny Brir.k 12 I OZONE INAC7IVATION OF IIUMAN o,-PROTEINASE INIIIBI7OR Previous reports have suggested that osidative components of polluted air, such as ozone, may inactivate .,-proleinase inhibNor (n,-PI) and perhaps lead to uncontrolled proteolysis due to inadequale concentrations of active k.. hibitor on the airspace side of the lung. This invesligation re•ea.nsines the i.- aclivltion of .,-PI by ozone so as to determine the possible involvement of this pollutam in the pathogenesis of pulmonary emphysema. Resuhs show thN in both plasma and pure inhibitor sduliorn, ozone decreases the Irypsin, chymo- tryp+in, and elastase inhibitory activities o/ human .r-PI in a concentration- dependent manner. The ability to inhibil human leukoerle elastase is also lod. In adddion to osidi:ed tyrosiee and tryptophan, inaetivNed a,-PI contains (our methionine sulfoaide residun, whose formation /rom methionine appears lo be the mechanism responsible for the inhibiloi s inactivation. As further dano.- strated by ekctrophoretic anatysis, the o:one-inac/ivaled a,-PI does ao/ form normal co.npk.es with aerine proleinasa. I1 is wqested thal ozone idtalaliota could inactivate n,•Pt on the ainpaee side of the lung 1o create a localized a,-PI de0ciency, which might contribute to the developrnenl of emphysema. /OAnron. D. Arnrrk.n Revltw o/ Respiratory Distett 121(6):10)I-10)S, 1980. From the Department of Biochemistry, East Tennessee State University Colkilie of Medieine, Johnson City. INACTIVATION OF BRONCIIIAI. MUCOUS PROTEINASE INNIBITOR BY CIOARETIE SMOKE AND PIIAGOCYTE-DERIVED OXIDANTS To test the possibilily that cigarette smoke components couW inactivate bronchial mucous proteinase inhibilor (BMPi) /n rU.o, BMPi was Mcubaled with an aqueous solution of fresh cigarette sewke and, after 30 rniwutes al )7'C, the elastase inhibitory capacity (EIC) was determined. ResuMs sbowe~ that there was a differetsce between control and smoke-trealed reaction wtis- tures. tw the absence of smote, conspleses of OMPi and elastase were present. whereas free enzyme was nut. By conlrasl, in the case of smoke-Ireated asia- tures, compkses of BMPi with elastase were decreased while free elastase a.d uncompksed BMPi could be detected. Antio><idants prevented this eAect. Chemical osidants (Nehk.rosuccimnside and ctsloramine-f) were also shos.n to ina.yivale the 1:1(' of BMPi. Iw addition, the phagocyte oaidiiing systern, myeloperoaidaae I a source of 1/ O, 4 CI , also suppressed the EIC of BMPi In vlnu. Similar results were oMained when the closely relakd human seminal dasma inhtbitor-1 was used in place of BMPi. Fin.lly, the functional aetivity of BhlPi was significamly reduccd in tracheal ay+irates of human smotaers compared to that of noesnsukers. l he results reported here support the hypothesis that local inactivation of BMPi in the conducting airways of the lung by inhaled cigarette smoke or by phagocylederived osdanls may play a role in the pathogenesis of obstructive lung disease. Carp. If. and lanoff, A. Esperintrntaf l.ung Rrfrarth 1( 7):'-21-II7, 1990. )31

1 Otb.r.upp.rta U. S. Public Ileahh Service. Frons the Depntmenr of Pathobp, State Universily of New York at Stony Rriwdc, Stony Brook. APIiD CI:I.IS AND NFIiROEPITHFLWL hIODIFS IN IIAMSfGR LUNG: MtINODS, QUAN111AT1UN, AND RESPONSE TO INJURY Three different hislochemical n+Nhods for the staining of neuroendocrine celk were compared in order to determine the mo.l c0icient techniyue (or demonstrating these ceRs. OI these, a modified Ea1on-Fedde procedure proved to be the rrwst efficient arNl easiest nudhod. With this particular lechniquc, the normal hamster lung was found to contain a total number of neuroendocrine eelis ranging from 2.0 a 10-'/tnrn to 3.0 t 10-'/mm in the srnall and large bronchioles. In the larger airwys, about ).31 a 10-' neuroepitAelial Modies (NI'Rs)/mns were observed. Immediately alter 2/ hours of e.po.ure to NO , the number of APl1D (amine precursor uptake and decarhosylat«rn) ctli, dropped to aboul 25% of the control values. After 14 days of e.pmurc, their number had risen to about 50% of the control kvels, remaininp at about that point until day 28 lhere was a Iransient drop in the number of NF.Rs after 2/ hours of NO, eaposure. MN this returned to normal by day 11. lhe best procedure for the detectw+n and analysis of APUD and NER cells seems to he the application of fluorescence techniques with standardized sections and quan- /rtalive methods of study. Palisano, I R. and Kieintrn..n, !. T/ioraa )S(S1:)61-)10, 1980 Otfier supp.rts National Ileart, Lung and Blood Institute. Frorn the Depanments of Pathok+py nd Pathology Research, S1 I.uke's lld- pilal, Case Western Reserve Univenity, Cleveland; and Mount Sinai School of Medicine ol Ihe City of New York, New York. INCREASED RRONCIIOAI.VFOLAR IgG-SECRETING CELI.S IN INTERSTITIAL LUNG DISEASES An Irnmwalopic basis for intersliual lunp diseases has been suggested by the frequenl anociatiow of wtoimnwne phenomena, and the increased rmmtber of inflamnnlory cells and proteins (especially imrwunoRlolwlins) found in the bronchiodveolar-lavage fluids of these patiewts. A reverse htmnlytic plaque assay, which measures cells that actively seerete elau-apeei(k immunoplolwlin, and hs use so deNne hnwwtK+plobulin-secre/ory function in normal lunp have previously been described. Reported here are the marked increases in geomelric tnean eahrea for IpG-seeretinp cells found in the bronchoalveolar lavage fluids of 22 patients with actire interstitial (unp diseases. Based on clinical evidence and on resuhs of linue eaaminalion, the group was divided into three cate- gories: sarcoidoris, idiopathic pulmonary flbrosis, and miscellaneous interstitial hmg di+eases. AN fwt two patknts were untreated and had clinically active disease at Ihe time of the initial study. Four individuals with non-nleralilial lung disorders and 19 healthy volunteers acted as conlrok. In those patienls who responded to corticosleroids, the bronchoalveolat 1g(i-secreling cells gen- erally decreased to levels in the normal rance. Conversely, they remained very 34 numerous in those who did not respond to cotticostcroids. The data, therefore, suggest that the numher of hr.rnchoalvcolar 1&G-secretinp celh reflects disease activity. Conaequcntly, this type of mcasurernenl may provide a more sensitive assay of such activity Ihan the currently available clinical indeaes, and snay prove to he a useful clinical 1oo1 for assessing the therapeutic response of patients with inlentilial lung disorders. L.ewrtnrt. F. C. rt .f. Thr Ntw F.n0and )ournd o/ Mtdirine )02(21):11lZ6-11811, 1920. Otbtr aupportr American Lung Associalion and the National Insiilules of Ileahh. From the RockweN-Keouph Pulmonary Immunology Laboratory and IAe General Clinical Research Center of the Melhodisl Ilospilal, the Ilarris County IlmpAal Diatrict, and the Veterans Administration Ilospital, Raylw College of Medicine. Ilouston; and the Metabolisnr Rranch, National Cancer Institute. National Institutes of Heahh, Rethesda, Md. ANAI.YSIS OF RRONCHOALVEOLAR LAVAGE IN NORMAI. IIUMANS AND PAl1ENTS WITH DIFFUSE IN7ERSTITIAI. LUNG DISEASES The results of studies on hronchoalveolar /avage (RAI.) fluid obtained from normal subjects, both cigarette srnokers and nonstnoken, are described and discussed in terms of the usefulness of RAI. analysis in several groups of patients with interstitial hrnp disease. In heahhy, normal, human subiecb. RAI. is a reproducible procedure that provides a generous amounl of airway proteins and respiratory cells. IrwkpendeM specirwens fntnn opposite pulwrosary lobes yielded ahtwsl identkal rewhs, wppesliep /ha1 subseproenlal lavape oRers reproducibk assessment of the other airway surfaces in healthy Wnpa Y1r paients sekcaed for study suffered tran two types of iwlerstilial disease: (1/ thow elicitinp a pranulo/natous histologic ttspowse itt hwy parenchyrna, iw chdinp estrinsic allergic alveolilis due to i.haled protein aNipear or spore , sarcoidosis and cosinophilic pranubnu: awtl (2) idiopathk MlentilW paer- nwnitis and fibrosis (IPF). ('barac/erislic t•AL patlerns were found in bolh normal and abnormal subiects. Active pralnAorwNous d'nease was assoeiNd with a distinctively high pereentape of lymphocrles (predominantly T-ceM). An increased percentage of polymorphonuckar panulocples, a few eosinopMk high kvels of IpG, and imrnune compleaes were indicative of IPF. SipniAc.nQ2. nareover, the RAI. da1a, s'plliun indes, an/ the imrnune eomplea lilen correlate well with the degree of parenchymal eeNula'.i1y iw lung biopsies and thus permit evalnation of the local disease adivNy. In IPF palients, serial RAL analyaes may provide a sensitive parameler for assessing the activity of Ihe disease in its later stages, which can help to determine rhe appropriate therapr. Rtrnoldr. N. Y. and Merrill. W. M. INSFRM 11:227-2t0, 1979. Other support: National llearl, Lung and Blood Inslitule. From the Pulmonary Section. Department ot,lnternal Medicine. Yak l)niver- sNy School of Medicine, New. Ilaven, Conn.

I I I FREE S1=('RFIURY COMPONENT AND OTHL'R PRO7EINS IN IIUMAN LUNG I.AVAGE Changes in the rne of synthesis and composilion of Rlycoprdeins found in rat tracheal eaplants esposed to ozone wggesl /he possibility thal epithelial proteinaceous products may re/lect celhrlar injury. This study anal)zes human lung lavage fluid for free secretory eornponestl, a Llycoprotein syn hcsized by the serow cells of the bronchial epi/heliuns. Subseamenul bron:hoalveoiar lavage (8A1.) of the lingula and right widdla lobe was performed on normal suhlects (l2 cigarette smokers and 24 aortsnwken) and the recoverable fluid analyzed for its content of respirMory oeps, blal praeins, .lburnin, 16(i, IRA, and free secrelory component. There was a oowsislenl increase in the nurnAer of cells, primarily alveolar mactophaps, contained in the lavap fluid re- covered from the srnokers. Polymorpho.wckar granulocylea were alto 10 limes more numerous and the total number of lymphocyles recovered was ithout twice that found in nonsmokers Protein analysis supported previous nw;res rn that the SAI, fluid recovered from both snwken and nonsmokers di;played an ekvated IgA:aRwmrn ratio when compared so serum; only smokcrs showed a higher 1g(3 a1hM.mm ratio. The recoverable mwuM of secretory romponenl was ngndkamly decreased in the young smokers and, wrprisinglr. Ihis did nnt correlate directly with pack-years smoked. Tlun, secrelory, ocomponent ays- Ihesis may represent a+ independent biochemical marker of epahelial dys- funcrwn, and may consequently be a sen.ilive Indrealor of bronchial epishsllal cell function and of early inhalant induced injury. Merrdl, W W,(ioodenberger. 1) , Suober, W.. Mallhay, R. A., Naeael, (i. P., and R.ywolJr. II Y Areerkan Revitw n/ R.rprarory Durase 122:156-161, 1980, OsAer aupp.rtr National lleart, Lun4 aad Blood Institute. From the Pulmonary Secrion, Yale llniversily School of Medicine, New Ilaven. Conn.; and the ( lrnical Immunology Section, National Cancer Inaluute, Na- l'anal I.s/Nules of Heahh, Sethesda, Md. PULMONARY ANAPIIYLAXIS AND THE KALLIKREIN-KININ SYSTEM Sisce its discovery, bradykinin has been thought lo have a role in acute in/lammalion, bul its precise involvemenl (or Ihal of one of its higher honw- Mp) in the pathosene.is of anaphylasis in Reneral, and of pulmonary ana- phylaais in partrcular, has nol been defined. lhe studies presented here had three ohjectives: to (1) esamiee blood-free hnK lissue for a kalbkrein-like enzyrne; ( 21 develop a simple means of moniloring the release of kallrkrein-like enzynles by anrigenic challenge of sensitized lungs; and (7) determine whether pre/realmenl of sensitized gu'rrea pigs with a kininase 1/ (angiolcnsin eon- vertinR enzyme) inhibitor irnproves or wonens anaphylasis. 1?saminalion of hrnR homoolcnNes revealed evidence of one or rnore kallrkrein like enzymes and a saae of 1inrno`en /lnwever, rhere was no sign of any rekase of kallikrrin like activity rnro the venous eRluent of anaphylaclic lungs perfused 16 I with KreAs-/lcnsekil solution; swx was there any release o/ kininase 11, an endalhelial cell enzyme. Induction of awaphylaais did not produce any chwoes in arterial kininase 11 kveh, showing that this enzyme is not a reliable kndea of lung capillary injury. These results do no/ attppon the concept that kaNikrei. rekased from lungs participates in the cardiovascular collapse of anaphylaais. Whelher or not the kaMikrein-kwwn system plays a role within the lunp, hosr- erer, is less clear. Neither of two drup that caw poleswiale the actions a( kinins. i1PP„ or SQ 14,213, had any effect on anapbylaais for better or for worse. 11 is still not known, however, il Ihese agents caw be delivered elBciewtly to lhose pulmonary sites where tsusl cells are i. the RreMeyl abundance. Ili6h doses of aspirin or sodium sdicylate can offer complete protection apinsl increased insuRlation pressure ud deveiopmeM of IurtR hemorrha6cs, the major pulmonary chanps of anaphylaais. Aspirin N lower dosage, a.d indonselhacin at all dosage kvels Iried, greatly enhanced she effects of anaphylasis on lung structure and /unction. To define how high dosage kveh of aspirin uwsfer their protective effects is the present challenge. Oza. N. B.. Ryan. l. W.. Rran, U. S., Serryer, P., and Pesa, G. AJvanerr in Eaperlnwnral MrJieinr and Qiolopy 120A:47)-4a6, 1979. Other support: U. S. Public Health Service, Hartford Foundation aed 1!a American Ileart Association (Palnt Beach County Chapter. Inc.). From the Department of Medicine. University of Miami School of Medicine. Miami, Fla. CYTOCIIALASIN 8 AND TNE STRUCTURE OF ACTiN GELS Cylochalasin R is a tunpl melaboli/e that i.hibils many cell wurewrc.h. Analysis of the 6A slranure formed whew tsucrophaRe adin-6irrRnQ proleia crosslinks skeletal rwwck anin Polyeters, astd eaaminaliotl of the eQec1 d cylochalsin S ow this structure srpport the oowcepl Iha/ eyfoplasrnk sol4el Iransformalions have a si6niAcanl role is cell physioloRy. Mcauremea o/ dr nclin filament length dirribulion has nsa.le iN possible 1u eaktiale IAe deo- relical critical concentration of crosslinker reatrired to indttce acdft polymer plation. The esperimenlally dctermined concentration reauired agreed with the theoretical one sufliciently well to asechrde that F-aelin:anitrbi.dip protein networks are isolropic Rels whose riRidity is regulated by acissiow of Rlamen/s. 7 he following effects of cytochalasin II on the ac4n nesworks O1 Ihn makl: (1) bndin6 lu F•aclin, but not to actin-batdinR protein; (2) de• aeaud aclin filameM length without an increase in the quantity of nsonomctic actin; (3) decreased riRidity .r/ actin networks, with or without ero.sGnkiq proleins; and (4) increase in the critical concentration of aclin-binding pro• Iein required (or incipient grlatirsn by a lheorelically predictable mallnNUde. 7he effects o/ cytochislasin 8 on plalion depended largely on the actin eon- eenlration and were inhibited by ph+Noidin, an aclin-stabilizing agent. It is eoncluded thal cyaahala.m B weaL.n, the actin 4e1 structure by severing aclin filaments at limited sues. Ihcrehy, rapidly reducing the aclin bindinR protein xtUn liiamenl ra1NS. 11k set s.d Iransformations eaused by such 37

hinued cleavage in actin networks suggest that this may be  hithcrlo un- recognrzed powerlul control mechanism lor altering cyloplasmic consistency and, thus, structure. llartwig, 1. If and Srorrel. T. P. loarw.l o/ MoJrrula, lioloty 1 II:S)9-Sf), 1979. Other a.r'ort r U. S. Public Heahh Ser.iee and the Edwin S. Webster Foun- datios. Frnm the HemaasloNy-Oncolop Uei1, Ms.achusats General Hospiul, De- partmenl of Medrewse, Ilarvard MeAkal Sehool, Bnston. hISTRIBUTION OF ACi1N BINDINO PROTEIN AND MYOSIN IN MACROPIIAGFS Itl1RIN(l SPREADING AND PHAGOCYTOSIS Spreading and phagncylnsn by macrophages are activities that involve movements of the corsKal cyscy+lasm Actsn lilaments- which are the prin• cipal suheel/ular strucsures vitd,k rn eketuon mrcrographs of the cortical eytoplasm, scem to have a/unctrors in these motor acuveres Achn budsng protein (ABP) and myosun lorher macrophqe proteinsl interact with actin In .Irro to wsAuence the Afusscnts' rigidity and movemenl, respectively. In this papn, the distribution of ABP and myosin molecules in acetone Ased rabbit IunA macroplsaAes was esarss/ned by wsear of immunoAuorescenee. The stain- in= for both of these proteins in uaspread eeNs was quile uniform, hut was reduced In the nucleus and corscentraled slightly ia the periphery. The peri- pherd acewnulation of staining attenuated in uniformly rpread celb, ahhough Alopwfia and hyaline veils were dcAniley stained. Irt ceNs Ascd during iss- geslion of yeast particles, the brightest staining correlated with the disposi- tbn of organeNe eacluding pseudopodia Mitiaqy surrounding the yeast. After phaAocrtosn was completed and the )easb resided in intracellular vacuoks, no eoncewlratioo of s/aiwsng around the ingeaed yeasts was ddectabk. llris indk.tes strongly that ABP and myosin nwkcuks are components of the hydise corte., structural unit of the ceN responsible for spreading and phaAo- cytosis; this is a regiors known to be rich in actin Alaments. lhe Andinp here are eomistent with the theory that lhese mokcuks control the rigidity and asovemenl of Alaments in the periphery of the li.ing maerophage. Srend.M. 0 1. , tlsrlwst, //1 , BroMcbe, E. A., and Sronel. T. r /owreal nl ('.ll Au+fugr M1/21 215 221, 1910. l rnm the llam.t.,logy ()m..logy Unn, Massaehusetts (:eneral Hospilal, and the Ikpaimcni ul Medicine. 11.rvsrJ MeJKal Schonl, Boston. t TIIE FFhI:Cf OF 111E OXIDIZIN(; AGENTS CHLORAMINE-T AND CI(:ARETI E SMOKE ON DOG SERUM PROTEINASE IN111BITOR(S) Under alkaline condition., chlorami.e-T (CT). an oaidkinp apenl, can modify methionine residues quite selectively. 'iltis report establishes that doR serum Irealed with CT in vitro is rapidy ..d selectively depklcd of its iw- hibitory cifect on porcine pancreatic daslae or dog wculrophil clasttue. The serum's trypsin inhibilory capacity is .01 affected. Puri/kd dog .-1-proleinne inhibitor (.-1-PI ) is aho osidized. Dithiothrti/d reverses the oaidali.e inactiva tion reaction. ('igarette srssoke also directly affects the irshibilory capacity of both serum and pure .-1-P1. Dw in a twawner wryerinA that it does not act Itw1 by oaidizinA methionine proupe. Theso studies densowurak Mat iw t doA,he a-1-P1 is readily inactivated by an oaidiziwg agent that dso aAeeta meshiortine residues. This reaction can be reversed by a strong thiol reducing qewl, a.d the cl.aase inhibitory site-ib rwort sensitive thaw /he tryp.is one. loAelher, these results are the baab for aw /w .i.o wadel of decreased functional .-1-PI levels that may prove valuable iw Ike study of awphysema. Abrarru, W. R., Eliraz, A.. Kimbel, P., and Wdwl.nrn, G. E+rrrinwna/ Lenp Rese.rrA 1()):211-22), 1980. Other a.pprtr National Heart, Luwj and Rlood Luituq. From the Pulmonary Disease Section, Albert Einstein Medical Center, PLila- dclphY. IMMUNOI.OGIC LOCALIZATION OF ELASi7N BY ELECTRON MICROSCOPY Much recent work ow the etiolop snd pabAeatsis ot eaperinscwtal ew- physema has been directed loward electron microscopy studies ol the dirribtr- tiom atsd deslruction of elaslin i. tke iMerstiti.l spaces of the alveolar waNr. Since the ill-deAned orpnization of elasliw in the alveolar iMerstiliun4, p.r- ticulary in the emphysematow IILA, t.ukes il di/l'icuq to identify the datio speciAcaNr by the staining procedures ordinarily used for ekctros ssicro.copy. a new technique usinA hiAls-titer anlibodies apirnt puriAed canine htwA elaslin was developed. llre esperimenls reported here demonstrate that speciic anti- body against insoluble dog hsrrR amorphous daMin can be used lw a staining procedure to localitt dog IunA claslin either by p.cembaWiwA or poaembedl dwsil procedures for ekctron miRoscopy. Loealiiation of the aMklastiw IgO wn accomplished with lerrMaalabckd rabbit anlisheep IgG as the secondary antibody. Trealmenl with the preimmune serum pre negligible /errNis back- Around staining. The anlielastiu anlsbudy did tal react with lur.p connective tissue poteins such as the mkroAMillar component of elasdw or eoNWw er proteoglrcan. Mast irnportanl/y, the antklastiw awtibody appeared to be species speeilk. 77se date presented in this paper establish the feasibility o( using .n immunologic approach for the iwkntificatiow d/he amwphous cornponent o/ canine alveolar interstitial elastin. 1 hi. techniQue could now be eapanded ly ,u

to identily and localite the microflbrillar component of etasuin snd other structural proteins within the hmt. Damiano, V. V.. Tsang. A.. Christner, P.. Rosenedoom, l., and WrinDounr, G. Arnrrtr.n Ior.n.d o/ t atAofogy %:1)9-156, 1979. Other nrrportr National Inslitutes of Health. From the Franklin Research Cenler, Philadelphia; the University of Pennsyl- vania School of Dental Medicine. Philadelphia; as+d the Alberl Einstein Med- ical Center, Puimosary Disease Sectiors, Philadelphia. STUDIES ON REACTIVITY OF /iUMAN LEUKOCYTF. EI.ASTASF, CA711EPSIN O, AND PORCINE PANCREATIC EIASTASE TOWA{:D PEPTIDFS INCI UDIN(i SEQUENCES RELATED TO THE REA(TIVE SITF. OF a,-PROTEASE tNH//11TOR (o,-ANTITRYPSIN) 1lurnaw kutocrte (Ilt.) el.swse and caMepsin Oore two serine pro- teases which have been implicated in the proteolysis of lung elastin olnerved is pulrwonary emphysema This paper reports a,tudy of the reaction of HL elaslase, cathepsin 0 and porcine pancreatic elastase with a series of qeptide substra(es. Here, the enzymatic hydrolysis rstes were followed by rneans of a ninhydrin assar using in each case the hydrolysis product as the stitndard. The cleavage site of aN .the peptides eseept the odapeplides was invasipted by thiw-layer ekctaophoresis Results sho.w that, amont other thinp, most of 1he peptides had a prolyl residue at P,. HI. elastase prefers a valyl residue over Ala or Met at S, while PP elastase prefers an Ala. With both el.stases. ettensioo of 1he peptide chain resuhs in signifkant increases in K_,/K.. With the appropriate subM.Nes, HL elastne is as reactive a PP elastase. CNhepsi. O shows a preference for Phe o.er Met N S, and a preference fo. Phe over Ala or 1 cu at S.. The .,-protease inhibitor (.,-PI) reactive site has recently been shown 1o have the aeryuctrce-Ala-Ik-Pro-Met•Ser-Ik-Pro-Pro, where the aslerisk indicales the bood cleaved (P -P,') when a-Pl-protease compksn .re aplil. In this study, the octrpeptide Ac-Ala-Ik-iro-Met-Ser- 1k-Pro-Pro-NHr atd the uwdopse with a P; 7Ur inslcad of Ser were qrrahe- sittd. AN three entrmes bound to and cksved the peplides at the P, Mel bond. The vahres, a meraure of the ease with which certain types of bond formation between proteases .nd .,-PI would occur, were higher tor the P,' Set oelapeptide /han for the 7lr analogue. indicalin6 that relatively minor .mh.o iscid suh.titulions in the a,-PI reactive sNe wip profoundly infht- e.ce i1s reactFdtlr toward vario.w Prole..es. Further audies indicate that re- .Nive dM subwitutions could also aAeet the degree to which o.idation M damaging to the inhibitor and may be om posaible explanalion for the greater susceptibilily to emphysema of some individuals with normal .,-Pl. McRae, s, Natajlma. K, TrarOr, I. and Powers, 1. C. lrloMrndu.r 19( 171 1911 1171, 1910 I rnm the 4h,.n1 .d 4 h.wu.uv (%.o.t,a Institutt of ie.hnol.,ty, AdaMa, and ,he (4panrnrni .4 fl.w A.....uet, lf.o.....it, of (icort.a. Athens 111. /Pe.rt .nd Cireulation SOME ASPECTS OF MYOCARDIAL METASOLISM DURING ACUTE INFARCTION New insights into myocardial metabolism have led to new approacha in the treatment of heart disease. In the paper presented here, an attempl is made to correlate the met.bolic lndinp with the clinical effects of eee well- studied heart disease, acute myocardial infarc/iow. Various sludks have siown that the generalized metabolic stress of acue myocardid Waretbu ekvMa free fatty acids (FFAs) in both plasma and myocard•+d liswe. Thia increaae, which is due primarily to the release of norepinephrine, inhibi/s Rhacose osi- dation by rnusck and uuses an accumulation of FFA in the heart nmsck. Also. FFAs or their acyl derivatives inhibit phosplw/rudo~inase. Therefore. FFAs or their acyl derivalives .re rae-/imiting in the Rlpcolytic as well as in the citric acid cycles. TLis results is diminution of ntyocardi.l pertornr ance. Acwe myocardial 'rschemia prr ir leads to an e.cess of reduced eauiva- knts, and inhibition of rate-limiting erwymes in the glycolytic cycle. The severity of ischemia determines the estent of myocttrdial damalle. Afso, after resumpimn of coronary Oow following myocardial echemia severe damage /o 1 mitochondria in heart muscle (reper/usioa syndrome) is noted. This may be the resuh of Ca t t awd/or Or deprivation followed by their sudden oversrrp- plr to the heart. The degree or duration of ischemia Is of importance in deler- miaing the sereriy of the damage following reperfusion. The clinical hnpli- eaions of these Ilndinp me discussed with special reference to the effects of beta-blocten, polassium-imulin and Ca f f aMago.ists. IJnp, R. !. aad Weishur, R. In: Proceedings of the FJorenre Interrw7ond Meet/nr on Alyocrdid In/.rc- tlon: Escerpa Mcdica, May 8-12, 1979, vol. 11. pp. 515-524. OtAer wrprt: Hoorer Foundatio.. From Iluntingtoa Memorial Hospital and the Huslingto. Institute of Applied Medical Research, Pasadena; University of Southern Cali/oraia, Los Aalieks; and the Department of Chemistry awd Chemical Engineering. Cdifornia 1.- aiNMe of Technology, Pradcu. TOXICITY, PHARMACOKINETIt:'Sr AND CHOLESTEROI--IN/lll11TORY EFFECT OF 7-KETOCIIOLFS7 EROL An analogue of cholesterol. 7-hetocholeslerol, inhibit@ cholesterol Aut iNo the walb of anerics perfused in rGro; it also i.hibits eholeslerd Sua in the arkri.l wall of int.ct animals. The present study eatuniaed the bsieilr and pharmacokinetics of this o.yteeated sterol in rabbits. Iw the toakNy studies, hislolop, hematology and blood chemistry were compared in four control animals, three animals receiving high doses and four animals reoeiv 41 All

ing lower doses of the osygenaled slerol. Each animal received a tolal of 16 injeclions at the rate of two injectrons per day. Pharmacokrnetic studies on the disaN+earance rate of (4-"(')7-KC were also carried out. Results showed that the histobgical, hemalological and biochemical changes in the animals es- posed to various dosn of 7-KC in which some 6ranulornalous an6iitis in the lung was seen Contrary to eapectations. the liver showed mostly fatty changes, which were also observed in the control animals. Inhibition of arterial flua of choks- Ierd (inhibition of 55%) wr swrcd with both high and low doses of the oaidited sterol. The disappearawcs c+rrres of ("C)choksterol in blood and plasma were characteristic of a 2-compaslteeM model. The rate constant de- termininR tnsue uptate of 7-KC wr higher than lnsue eAlua and there was no appreciable reflua into red ceMs. These resulta indicate that it is possibk Io reduce choksterol fius in the arterial wall of rabhiH without causing major to.ic changes and that both red cells and tiasue act as a reservoir /or the osygenated sterol. Santiltan, O., Sarma, 1. S. M., Pawlik, O., Racl1, A., Grenier, A., aod Sing. R. l. AtAeroKlNoliJ JS:1-1O, 1980. Oth.r anr/.rf r The Hoover Foundation, Anxricaw Pgg doard, and the U. S. Public Ncaltls Service. From /IuNirgton Memorial Ilo.pital and Huntington Institute of Appfied Medical Research. Pasadewa, Cal ; Uwi.ersity of Southern California, l.os Angeles; and California Institute of Technolop, Pasadena. Tlle EFFECT OF REGIONAL MYOCARDIAL ISCHEMIA ON SERIES ELASTIC AND CONTRACTII.E ELEMENTS OF GLYCERINATED HEART MUSCLE IN DOGS Many fadors contribute to the deressiow of cowtractility Ostter sstTs eardial ischerwi., but relatively few iw.erriptiows have focused ot the oo.- tractlk Ts/em NseN. This report don e><amiwe it, wi11r particular ewrph.aM ow dm acti.. Mif/wes.. Isomtuic eoalrwctiow awd quict release me%hoss wers nsed to snrdy the response so hchernia of the eonlractik wwd series elastic ekmeMse owce the passive knph-lewalow relationshiP iw alycerinated heart wwt.cle Rhers of dop iw the restisy Mate had beew mesw.ed l.i6atiow of the left coronary artery iwduud snyoeardial M/arct{ow. Muscle Rbers from the ischesnic area talew three hows a/'er ligation demonstrated reduced ma.imd devebped sewsiow (P.), masirnal rak of lanion development (dPrt...) wwd V.,. The time to peat sesssion, 1„ ahw iwcrened. As occurs in fresh papil- lary muscle, the modulus of elasticity of active glycerinated mus:k rose in proportiow to Wad. The stillness of the series elastic element was significantly elevated iw iachemic muscle Rben. Iw iacchemie heart sswsek. the p.ssive sti//- ness al the restinR sta/c showed a decrease in the dope along with aw increase in the intercepl. The esperimeata show thM, throughout the vsrious load range used, dnrurbances in the contractile element occur when there is in- creased strflness of the series elastic ekment without significant merease in p.ssive atiflneu. Some relalionship between the series elastic and contractile ekmenta is therefore suggested. Maruyama, Y., Fischer, R. and eing. R. l. l.p.nere Cirrul.rion /oavwd 44(6) :4N-460, 1980. Oth.r surPorfr Hoover Foundation and The Wright Fouwdatiow. From the Huntington Institute of Applied Medical Research nd the HuntiwR- tow Memorial Iloapitd, aed the California lwstitwe of Tech.oloqy. Pasadeaa; and the University of Southerw California, Los Angeles. SINDIN(3 AND INTERNALIZATION OF LOW DENSITY LIPOPROTEINS MY PERFUSED ARTERIES Studies oa cell cultures dealing with rhe eRect of temperature os binding and 'wHerwalizatiow of IowdensNy lipoproleiw. (LDL) have been publishd before. In the paper presented here, the eAect of low temperature (4'C) aw Irypain-rekasabk and trypsin-resirawt 1.DL wr investigated fw porcine ar- teries perfused with puhutile pressune. Perfusion at 4'C resulted iw a siR- wilicant reductbn in arterial LDI, uptake. Sigwificant diQerences were fowd alw in arterial LDL uptake alter treatmewt of the aatery with 1ryp.iw. Ow the other hand, the wmourM of 1'sa/1-IAL releued rrons the weteries did wol significantly diAer under Ihex circumstances. There/ore, lhese observations strongly suggest that in the perfwsed isolated arteries, as in tissue cuNures, the anaunt of surface bound LDL is ww dlected by temperattrr,r, while iwtcrw.b Itation of LDL is martedy reduoed. S.Millaw, G. G., Schur. l.. Chan, 5.1.. wnd Iiwt, R. 1. /JocAenrkd .rrd eiopA)sk.l Reu.rrA C.wutrwwkardowr 9S(4):1410-141d. 1990. O/Aer .w''.rtr The Hoover Foandatiotfr Atnerka. t:U Roard and th. Na- tio../ Iwurtssln of Health. From the Huntington Memorial Hospital. HuwlinRtoa Institute of ApPlied Medical Research, and the California Iwstitwle of Techwologgy. Pasadew.. 1.(X'AI.I7.Af1ON OF FIBRIN CONVERTED BY 711ROMe1N FROM R':I.EASED PLATELET FISRINOGEN The subceNular localization of fibrinogen and Ilbiiwot ew-related twoleculw in Ihrombin-activaled platelets was studied iw order 1o elucidate lbriwogrsr involvement in thrombin-mediated platelet function. Tre.tmessl of pl.te" (10' cells/ml) with thrombin (1 U/rM) caused rapid disappearance of A riwo6en from the spslem, as measured by the Ianned red cell hemaahNi.atiow hhibition immunoassay, (TRCHII). Although twusse.wrabk by TRCHII. Iha twnw- bbrino6en nevertheless remained closely asso=ialed with platelet surface 4Z 41

branes l he 7 R('1111 delectabk material was recoverable from platelet pcllets afler centrifugation and plasmin digcstmrs of individual pclicts. lhese and other results of ad.Mional e.perrmenrs described here suggest that there may be a direct association between the fibrinogens released frorn platelets and platelet surface membranes lhis may involve fibrinogen binding or forma- Ian of 6brirw+gen fibrin comple.es on Ihe plalekt surface, or crosslmkinp of Obrrnogen Io platelet plasnsa membrane and contractile proteins. On the basis of the present data, however, i1 is also concluded that at least part of the membrane-assocuted Abrinopen in Ihrambin-treated platekts is in the form of flhrin or polymerirmg fibrin intermediates. Furthermore, white the e.act state of the membrane-assrrcialed bbrinoWn released (rum platelet by thnnnn hMn is yet to be determined, these same resuhs support the sua`eslion that platekt surface membranes are important siln for interaction of platelets and fibri.ose.. CAao. F C., n .f Blood 33(2).111-194, 1990 . t)tArr aurr.rtr National Institutes of Health. From the Center for Blood Research, 6oslon. ARiI?RIOVFNOl/S SHUNTS IN DII.ATF.D OR REPERFUSI.D CANINE CORONARY VASCULATURE lbe e.tent of bkad Aow through aneriovenow shunts in the do2 hearl was estima/ed here under normal condilions, during eslensive vasodilalion and during reperfusinn of aculely ischemic myocardium. Radioactive micro- spheres 77-10 rm iw diameter were used as irdicatots of eoronry shunt flow. In Group 1(12 dop), the kit eommon coronary arkry was cannulated and perfused with arterial blood as aortic pressure. Venous blood was collected from the coronary sinus. In Group IA (sia dop), the /ollowing measurements of percentage shunt Row were made: eorNrol, 2.0t0.2%; reactive hyperemia following 20-ac coronary artery occlusion, 1.5t0.2%; coronary dilation with adceosine, 20±01%. In Group 19 (sin dop), the following observations were made: control. I.S±0.7%, coronary dilation with adenosine and per- /usion pressure ekvated to 170 mm Hg, 2.4t0.3%: systemie hyposia due to inhalatios of nitrogen and per/usiow pressure elevated to 170 mm 11g, 4 2t0.9%. Only under the hypoaic condition was the percentage shunt Ilow silnilkantly elevated above the control condition. Values noted for Group /1 (sia dogs) did not diAer significantly arnong themselves or from the control observations in Group 1. The most importanl findings of this investigation were that only 2% of the kit coronary blood flow passed through vessela larger than 7-10 pm in diameter sad that this shunt fluw was nut elevated during reactive hypercmia, adenosine-irdueed coronary vasodilation, or teper- /usion of infarcung myocardium Only during systemic hypoaia with elevated I coronary perfusion pressure was shunt /low increased significantly, and 1hen only to an average of 4 2% of left coronary flow.. Downry, H. F. rr af. Alkrov.rurfar RruarcA 17•22-26, 1979. Other au'pert: American /leart Association, Tenas Af~iliate, I.c., a.d the Cardiology F nund. From the Cardiopulmonary Institute and Departmen/a of Physiology and Internal Medicine. University of Teaas Hcalth Science Center. Dallas. MYOCARDIAI. EFFECTS OF I.ONG-TERM CIGARETTE SMOKINO; RELATIVE ROLES OF CARBON MONOXIDE AND NICOTINE It has been establiaAed in an animal rnodal eaposed to lonp-1.rm ciprette arnoting that left venvicular performance declines iw association with rnor- phobgic evidence of eollagen accunwlNio.. 1o order to determine whether oiootine or carbow .wnoaide is the major pathoae.ic atleM in cigaretla ar.al., It tracheastomiud young adult tuak beaRla srere divided Into three equal groups. Group I acled as aowlrob; group 2 received ae.ea low-nicoti.e (0.2 wrg/eig) eonte.t ciprettea, which eAeded a peak carboayAernoillobi" kwl of about S/ 100 ml, airnilar to that of regular dprettes; and group 3 was Injected lwke daily with a. Maranwscular dmt of wicotiee equivalent 1o 1he amount is sevetn cigarettes. 1. the i.tan a.etthetiaed wNe, the 1Mee Rrayp did awt show any di0erewca is heart rate, kit .e.trkular et.d-diaMOlie pra- twe and volume (indicator dilution). 11VIw trlf.s aohNiwt was infused INtr the kft ventricular cha.nber i. order 1o assesm rdNive teyocardid waN ai/- .esr„ only group 3 de.eloped a ripiRcantly 4idsr eadJi.stolic preuun and Inuion, suggesting decreased kit .entrictrlar oowtpN.woe. twcreaaed kit .e.w trkalar myocardial hydroarproline ooatetri, wbkh may be a basia /or e.- \anced atilfness, was only associated with bty-lerwt t.icotine wage. As l.des of left veatricular contractility derived /rowt Me peak rate of left .eMrienlar pressure elevation (dp/dt), nornnliaed for prelo.d aad afterkrad, ahowed no difference for groups I and 2 at oimilar levels od aortic pressun, but wr signi6candy lower for group 3. Thu., the effects of long-term ciprette amok- i.g on the myocardium seem to be predomi.aMly due to nicotiM rather thao to carbon moaoaide. ~ Ahmed, S. S., Moschos, C. S., OWewurtcl, ll. A., and Rre.n, T. /. The Anmeriean Jorrnd o/ Cardiology 46( 4):393-399, 19110. Other.upprt: National Institute on Akohd Abuse and Akotolism. From the College of Mcdicine and Dentistry. New Jersey Medical Scbool, Newark. 44 45

CAPII.LARIFiS OF THE ADRIiNAI. CORTEX POSSFSS AMINOPI l' 17hASh A AND AN(i lU l ENSIN-CONV IiR 11NG- ENZYME ACi1VfT1ES 1 Ihis attempt to deAne the activities and cellular disposition ol angio- tensin-converting enryme and sminopeptidase A within the adrenal cortea, adrenal.gland cspJlary segments were isolated and prepared as suspensions. On the day after isolation, the cells wers assayed for angiotensin-converting enzyme snd aminopeptidase A activities. Sinwlary, dispersions of the adrenal cortet were prepared and assayed. Aq .ampks uad for the enzyme ass.ys were treated e.actly like the ones prepared from adrenal-ttand endothelial cals. Re- suhs of these studies showed that the amount of angiotensia-converting enzyme activity in the eedothelial cells (me•sured 60 minules after beginning the in- cubatiou) was apDrosimately si• times that of dispersrons of the adrenal coe- ses Aminopeptidase A activity of tha capillary seiments was 12 ti.nes that of the gland homoWnaln. A1 a And conceMration of 10 rM, the hrady- kiai.-potentiating peptide BPP„ (added three minutes before addition of substrate) inhibited the reaction of endothelial celb with substrate by 911R. The inhibitory eRects persisted In incubations of up to sis hours. OveraN. these results show that the enzymes required to convert the prohoemone anAiwensin I into •ngiotensins 11 and 111, secretsllogues of aldo+terone, are enriched in association with capillary endothetwm isolated from tal adrenal eortea. Thus, it appears that the secretion of aldoslerone may be wmrdkd, in pan, by the proeerinR of peptides occurring within the adrenal gland itself. Del Veexhio. P. 1., Rysn, 1. W., Chuq. A., sod Ryan. U. S. •lorArwrlrr Jorrw.f 186 :605 -60/, 19110. Odler sr'p rt: U. S. Public Health Service, American Heart Foundation (Florida Affiliate, Suncoast Chapter), aod the Hartford Foundation. From the Department of Medicine. University of Miami School of Medicine, Miamk FIa. MAMMALIAN INIIIBITORS OF ANOIOTENSIN CONVP.RTINO ENZYME (KININASE 11) A sinple, accurste and sensitive assay for anriotensin converting enzyme (ACE) is presented in this paper. The new assay measures A('FE using 1'111 Ilip,llis-Les as substrate instead of the (sHl Hip-Oy-Gly that was used in earlier studies. Results of these assays (enosty, new) showed that both blood and uri.e contain large amounts of sn1aM molecular weight inhibitors of ACP-. AhhouRh the winn (human. Auine• pig and rat) contai. ACE, the enzyme is scarcely measurable without prior uhraAhntion or dialysis The activity iecntases strikingly through three uhraAhration steps using a membrane with • 10.000 MW retention limit As implied, the ultraAltrales contain inhibilory sctivNy and can prevent the hydrolysis of /911/ benzoyl Gly-llis-Izu by ACE. 46 Human urinary ultrafihrate contains at least three inhibitors separable on Bio-Gel P-2. The inhibitors are acidic and can be partially purilkd by Bio- Rea 70 developed with an acetic acid gradient. The smallest of the inlwbitors cao be purified to apparent homogeneity by partition chromatography. In1er- estingly, the inhibitory activity varies in responsc to dietary sah: activity it low whea rats are maintained on a high Na(.'1 diet and is high on a low NaCI diet. Thus, the activity of A('FE may be modulated in viro by naturallyoccvr- ing enzyme inhibitors. Whether some hypertensive patients are defkkM i. ACE inhibitory activity remains to be determined. R7aa,1. W. rt af. (Ryas, U. S.) Advancra in EzperJmrnra/ Alydirinr and eJoloty 120B•599-606, 1979. OtAer .rpr.rf r U. S. Public 7/tahh Service and the Ilartford Fousdatio.. From the Department of Medicine. Universily of Miami School of Medki.•, Miami, FIa. STRUCTURF-/UC:TIVITY RELATIONSHIPS FOR KININASE 11 INIIIBITION BY LOWER HOMOLOGS OF THE BRADYKININ POTENTIATIN(7 PEPTIDE BPP„ The bradykinin-potenti.ting peptide, BPP„ ( <Oht-Trp-Pro-ArS-Ptu-Uhs- 1k-Pro-Pro) is a polerN Inhibilor of kininase 11 (nrRiotewsin convertinR ew- zyme). Several N-termiul, C-termind, and iakrnd lower hoawloRs d BPP„ have been synthesized and their kininase Il inhibitory potencies determined. 'iUe substrate hH/IlipHis-Leu was used to tweawre ki.in.se inhibitio., and the formatioo of 1'Hihippwie acid served so twe.wrre the enzyme. 71re• of the lower hornologs were found to he as polewt as BPP,,. The C-terwrind Iri- and tetrapeptides showed signi/kant i.bibitory activity, whereas the N-Ierminsl di-, tri- and tetrapeptides proved to be weak inhibiton. The <Ohr' serment is not indispensable for binding. but tnay protect the peptide against enzymic degradation. On the other hand. Tep' is critical to binding. Prdis as a penultimate C-terminal residue is generally unfavorable a1d pre.ewts or greatly reduces bindinA, but Pro' facilitates it. Also appearing to faeiliaNt binding is Ik' as the ('-terminal residue of a lower howrolog. The prese.oe of Arg• or of a basic side chain N residue 4 is not critical. As for Pros. Pros, and (ilns, they do not seem to be importaM for binding per sr, but may affect the overaR conformation of the peptide. Fisher, G. H. rt al. (Ryan. U. S.) Advances in Eyerlmentd Mrdicine and eiofotr 1208:631-662, 1979. Other arrprt+ Florida Heart Association and the 11•r1Fxd Foundation. Ftom the Department of Medicine. University of Miami School of Medici.e, Mi.wei, Fla. 47

. f .~f., 0 ~ ~ 1L~ m ~ c6~== r m m +l ~ N r tp J PREPARAl1ON Of= INIRINSICAI.I.Y-I.ARELFI) K1N1NS Although s great deal has been learned about the function of brady- kinin, little is known abou the mcchanism of its inaclivation This study was uarted as part of a program intended to help clarify the characteristics of tnzymes that inactivate bradykinia. SpecifleaBy, the goal ws to prepare a series of habgenated Madykinin andop, which upon catalytic dehalogena- ti+n in sHr gas, would yield bradykiwin intrinsically-labekd at hilh specifie rad'roactivity. Three analogs were sy.thesised by the Merrifkld solid-phase method and purifkd by molecular sieve aad partition ehromnokrahhy: 14-Dr- Phes/-hradykinin (raK), 14-Sr-Pkr%RK and 14-fir-Phe's1-faK. Tlse analogs possessed varying degrees of biologic activNy, as assayed by their effects on mean arterial blood pressure and isolated rat uterw. One of these, (4-Br- Phes1-RK, was uneapectedly found to be 1.4 so 3.6 limes more active thae bradykinitt. These results, when added to other previousiy prestnted data, suMesl that the interaction of bradfki.ia with its reeeptors, an.l probably with its metabolic enzymes, can be affeeted critically by the wature of sub- stituents on the phenylalanine riap. The (4-1111r-Phc's/-SK is ekarly resistant to metabolic dc4radation, while 14.f1r-Phe'1-SK is more ruknerabk. The 14-Br- Phe')-ftK, however, seenn to be melabolited at a rate similar to that of bradP- kinin itself, presumably by pulmonary enzymes. It is unlikely, therefore, that this analog derives its wperactivity (ro.n a resistance to metabolic degradation. The bromine sloa- however, may have favorable effects either on the c.rn- formarion of bradrkinin or on the relative dfinity of the analog to the re- eeptor lhese possibddres remain to be tested. C.huog. A er d(Ryon, U S) Adv.nrer Iw E.Pt.unrnro/ Mrdecw .r.d Aloloty 120A 39 50. 1979 OtAer aurprt: U S Publrc lleahh Service and the Hartford Foundation Frorn the Department of Medicine. llniversuy of Miami School of Medicine, and the Pap.nicolaou Cancer Research Irohtwe, Mumr, Fla HOW DO KININS AFFECT VASCULAR TONE? ANheu{h kinins and aupiole.sin are ckas.ifkd as hormones that affect smooth ntucM, it is by ta means clear law they do il. Previous studies have show. 1hM bradyki.ie can dilate pulmonary b/ood vesseM and greatly reduce pulmonary vascular resislance, whik other studies have shown that brady- ki.i. In satesivcly dqraded by enzymes sittutcd on 1he luminal surface of pulmonary endoaheli.l cells. Howeser, ahhoup/s endolhelial cells can inactivate kieiws during passap through the pulmonary circulation, the pu/monary vas- cular resistance still falls In respome to bradykinia. In thia attempt to under- mand the way kinins escrt their effects on pulmonary vascular resislance, a search was Nsrled for a morphok+kic basis by which tiniws, actinp on eedo- thelial ee11s, could influenc• coneraction/rela.ation of the adjacent smooth muscle larers Tn thtl end, election microscopy studies were done and pro.- tuglandie enduperucde synncctuse wss localized by autoradiography using 49 i ('H)acetyl-salicylate. Results of these studies indicated that endothelial ceRs and smooth mustk celb form specialized cep contacts. Myoendothelial jwrc- lions occur and smooth nmsck cells in pulmonary arlerioks send large num- bers of projections into the endothelial cells. In addition, smooth muscle cells attach directly to the abhwninal surface of endolhelial cells, as do pericytes. Thus, there is a morphologie basis by which kinins can aAec1 tone of smooth rnusck without acting directly on smooth musck cells. Ryre, U. S., Ryan, l. W- and Whilaker, C. Advancer in EiKrinrenrd RMdirinr rsI Qiolopy 120A:77S-391, 1979. Other wr'.rt r U. S. Public Ilealth Service and the Hartford Foundation. From the Department of Mcdicine. University of Miami School of Mediciwe, Miami, Fla. IV. Neuropb.ras.eoloty .wd Phy.iology DISCRIMINATIVE STIMULUS PROPERTIES OF THE OPTICAL ISOMERS OF NICOTINE The discriminative cue produced by subculaweous injectiw of .icollr depends upon the central rather Ihaw Ihe peripheral efiecas of the aRe.l on nicotine receptors. Structwal charrpes is the .iootine wsolecule decreasa ths dncriminahility of these compounds at compared to nieoliae, dertsowstratkeg the strict slructural requiremeM e( the wicwine receplor is mediating tbe nicotiee cue. The present audy was u.denake. 1o investigate pharwnoobpie stereospecifkity, another importaa reeeptor erileriow, in order to fwdher c!w- acterize the central nicotinic receptor. The abilitr of opicapy pure (+).Ka tine, the synthetic isomer, to te.eralizo to the eue produced by oplicaMy, purs (-)taccotiwe, the natural isawer, was drerefore lesled. Dosa-respew relationships for both optically pure isorners, as well as .nlagoninic eAects, were eaami.cd in rats trained lo dscrirni.Me either 200 or 400 p/kR (-)nicotine from sdi.e in a 1wo-bar operant paradigm. The natural Itomer was about nine tirnes mae poleM Ihaw (+)wicotine. MecamylamMs blocked both types of cucs equally. Ilssaered.onium and atropkse had no effect. T1esa data show that the central nicotinic receptor wrcdiating the slirmrlw eQtet produced by nicotine does have a dftree of s/ereospeciikity. Mcltzer, L. T., Rosecrans, 1. A., Acrro. M. D., aed Hanis L. S. rsrcAopAorNaro/osr 66/71:26)-2M6, 1980. Otfi.r .Mrr.rt r National Institule of Drug Abuse. From the Department of Pharmacology. Medical College of Virpi.ia. Rich- nqod. 49 1

SYNTIIFSIS AND PRFI IMINARY TIINOIN(i S1111)II:S OF 4,4 1)alkl1O.(-) NI(YIIINf 01: 111611 SPI:('lFl(' A('TIVIIY Since there is currently an interest in nicotine receptors in the central ncrwrus syslem, a need his developed recently for a radiolahelcd nicobne with high specific activity. lhis paper reports the synlhesis and some preliminary hendin6 shdies of 4.4 JeInto (-).nicotine, a compeMmd of high %pecific ac- livily (4 7('i/mmoll MhKh has been synthesired from (-1•nicoline via the readily prepared 4,4 drhromocotiawst. The distinct advantages of this sub- stance over commerci.lly available radiolabekd (--•nicoline are higher spe- cific activily, good stability, and IocMiow of the lrilium atoms al a specific carbon A Sca(chard analysis of the binding of 4,4-di(ritio-(-)-nicntine to the crude mitochondrial fracliow of whok rat brain revealed • K. oC 4 7 x 109 M' and 1) Imd of binding siles/rnt of prokin. Vincet, W. C., Mar(in, • R., AtNe, M. D., and sowman, E. R. Iournd of AItdJe-IwdCAtndrrry 2)(1):962-f64, 1920. Other aar"ert: ll S Public Ileahh Sertkt. From the Deparsmenl of Pharmacolop, Medical College of Vir6inia, Rich- mood AnRENOCORTICOTROPIN DEFICIENCY: CORREClION OF IIYP(1NAIRFMIA AND IIYP()AI.IK)STT:RONISM Wllll (-IIRONIC (il.U('(K'OR i I(-OIl)1 IIFRAPY A)6 year old woman, who complained of fatigue and progressive weight loss, eshibited a combined deficiency of adrenocorsicotropic hormone (ACT11), growth hormone (Gil). and prolaclin (PRL), although her levels of Ihyroid- sllmulatinR harnone, luteinirinp hormoet, and follicle stimulating hornsone were norteal. Aldoslerone eacreliow was markedly reduced (014 rp/d•y) before replaumem therapy was started, hw normal renin and aldosterone responses to sodium restriction were observed alter sis months of cortico- sleroid Irealment. In This pa1kM with the ACT)1. Gil. and PRI. deflciency, eortisol appeus to have beew The missinp lactor responsible for depressed akdoslerone secretion during the acute ilhmss. Its action on the reninaldoslerone system was slow: one week of phtcocorlicoid treatment did not immediately restore twr~nal sod'wm conservalion, but after six months of cortisone therapy, renin and aldos(erone responses had normalized and remained normal for several days after glucocorticoid withdrawal. It aeems, therefore, that chrow- icaRy •dequate ghrcocorsicoid levels were necessaty to mainlain a normal a1Jo- slerone response in this patient. If there is also a pituitary laclor required for this response. it does not appear to be ACTII. Mersiam, O. R. and 1arr, L. lotrrnd of Clinical Ewdotrlnolopy and MruAdlnn 30(1) :10-14, 1980. (lsher arprrl: Nalw+nal Insliture• of Ileahh and the New Ymt 'qrale lk- partmtnl of Iltalth Itirn he 11tN..nwm .d MtJ••.M. ( (+lumhsa Ilmwersdy ( ullegt nl Physleiam A •n.l It.np.isl Nt.r Yor- MECAMYI.AMINI: INt)IN'1?I) 01(X'KAI)F OF NI('OI INE INDUCED IN11I8111ON OF GONAI)nTROP111N AND TSN SECRETION AND OF NI(YlIINE INI)U('EI) INCRIiASES OF ('AlE('11(11.AMIN1? IURNUVER IN il1E RAT IIYPOTHALAMUS Nicotine may reduce LII and prolactin secretion both in the normal mab and the ovaricctomired female ra1 thrarugh activation of the tuberoinfundi- bular dopamine (DA) neurons and the median eminence. In order to deler- mine if these changes are related to the slirrwlalion by nicotine of central .icoline-like cholinergic receptors, the ganglioe blocking agent mecamylamine was used to see if N would inhibit the ho-monal and ealecholamase (CA) turnover effecla of nicotine wilhin the hypothalamus. Nicotine treatment (2 mg/kg in repealed iwNraperitoneal doses) of the ovarieclomired (emak rat significantly depkted the CA slores in various parts of the animal's media eminence as did the lyrosine hydrasytase inhihilot, .-methyNyrosine meshyksler (11 44/68). Nicaine enhanced the eRect of 11 44/68, but I. the torekain it did not singly deplete the DA stores in the olfactory bulb or the vesti- bulocochkar and caudale maki. It did, howerer, kssew the N 44/66-indueed depklion of t1A stores in the vestibuWcuchlcar tsuckus, if not in the aber two reRions. These samc drties of nicotine also markedly inhibited prdaclin. LN, 1'511. and TS11 serum kvek in the presence of 11 44/68. Mecamylamine treatment seemed to partly counteract the nicoline-induced inhibition of tolsa- dolropin and TSII secretion and, while it alone did not significantly alter hor- mone secretiots, it tended to increase '1 S11 secretion when combined with tha inhibitor. Nicotine also enhanced the 11 44/68-induced depktiow of the not- adrenaline (NA), adrenaline (A), and DA Nores in the normal male ra1 hypulhalamtn, while mecamylamine prelreatsnenl blocted the It 44/61-irtduoed ~ depktios of all the CA stores. Mecamylamint: alone enhanced the If 44/62- imhrced depletion of the hypothalamic DA and A stores and had a similar but not significant eRecl on the NA .rslems. sased on the evidence, it I. stall• R-sled that the DA. NA and A lerteinals within the hypothalamus an en- dowed with cholinerpic aiieoline-lite reeeplon that inhibit Ronadotropin and TS11 secretion in respoltae to nicotine. Anderssos, K.. Fuae, K.. Eserolh, P.. Gustdaton,l.-A., and ARnati, IF. Actr rlYyrlofopft a ScrnJlnrricr Suppl. 479:37-29, 1980. OtAer serrpsr/: Swedish Tobacco Co. From the Departments of Ilistolopy and Medical (-'hemistry, KaroNnsta b- uiltdet: and the llornane Laboraluty. Ikpulmenl of (1Mletrics and Gyne- cology, Karolios-a Ilospilal, Stockholm. MOI.1:('111.AR FURMSOF IMMtlN()REA('f1VE PANCRI:ATK' CAl IONIC TRYPSIN IN PANCREATITIS PATIENT SERA Acute pancreatic inAammaliun is generally thought to be brought abotr by inappropriate activation of the pancreatic aymosens leading to the rekata of active t -• --Te into the pancreas, pancreatic aecrction and the blood atrtarw. SiRnificanl', -cva/ed levels of itnmuoorcactive pancreatic Irypsin have been observed in the serum or plasma of patients with acule pancreatic alllamma- delermined 1 his communication describes The application of a radioimmuno- ~11 I s 1

o+ tn m m J ra W .n assay technique to Ihis problem. The averye serum level of immunoreactive ca"oak trypsin frons 70 patienls with .cale pancreatic inflammation was tion. Nowever, the molecular forms of this circulating lrypiie have not been 1.590 nR/mi as opposed to an average normal value of 26 a6/ml. Chroma- to4raphy. designed to resolve trypsinogen, ...ntilrypsin-bound trypsin and a; macroglohulin-bou.d trypsin and applied to serum samples from 14 pa- tienls, indicated in tach case that the major portion of Ihe immunareactive material was fret trypsinogtm while o0l7 a minor fraction appeared to be .,-antierypsin•bound rrypsis ConMwnlio, of 1he zyeaken nature of the maior peak was obtained in one east by activaliow witA hurnan enteropeptidase. Treatment of the .,-rnacrog{obuii. peak with acid yielded Immunoreactive trypsin in (1 of the 14 samples. R7 deseetlrtR eornpkses of pancreatic trypsis bound to .; macroolfobu/iw, it is hoptd that this tachniolue rnar help to eh.- cidase the pathophlnioloDr of tlds disease and tertaiw of its cornplicaion.. Srotlrkk 1. W., arol.r, M. D.. larkrwaw, C.. Passen. M.. awd lohron. 1. H. A.n.rkan Io..nd of rAyrloloty 217(S):E474-f?4R0, 1979. Of1rer .rp'.rl: Medical Research Ser.ke. Veterans Admi.iatrNio.. Fron" the Fauymobq Research Laboratory. Martinez Veleran. Aderinistra- tiow Medical Cewter, Martiner, Cal.. and Department of Mediciwe. t niversily of Califorda School of Mediciar. Davis. MOLFCUI.AR FORMS OF 1MMUNOREACTIVt' PANCREATIC ELASTASE IN CANINE pANCRP.ATIC AND PeR1PNERAL B1.0()D A r.dioimnnnroaasay for eaaine pancreatic elastase 2 ha. beew developed to define quantitatively both the pathway of eswry of Mw.unoreacti.e el.slase 2 into the bbodstrcarw .nd 1M n+olecular forms tbis rwa/erial eahrbita at st- lected poiw in the reewtr cirevlatio.. 71e present sUrd7 (ate.ds Ihe we of this /achdoqrre to an eaperitnewtal ea.i.e twodel that was used to in.ptiptu the twolecwlr /orwn of imnwnoreani.e panereatk elastase iw pnncretlic, mesesrteric, pnal, and peripheral rewow srnpks eMai.eA from L.N.R a,.ea- Ihctiaee dop before awd after sthwulNio, of pancreatic secretioe by cholecps- /okin octapeplide. Results show aka/ whe. periphenl renow ."i is (r.e- tbwNed ew Rio-Oel A-0.3 tw. a single peak of i.ww.ore.eU:le ctanase 2 h observed witli a molecular weight of .ppreaimately, 70.000, wAich a comis. tenl with du/ of a proelarast 2-a,-protene Inhibitor eompkt. In contrast, whee samples of pancreatic .enoua plasma an fractionated on Rl>Oel A- O.f w, two peaks of Mrrwrnoreacai.e elntase 2 ftatinR ...pproabnat relative ntolectilar weiillrt (M.) = 70,000 .nd 23,000, respectivtl7, are observed. Ae- Iivaiest with Iryptin has showw that the (aNer represents proelastase PoMow- inR cholet7MOkiain-octapeplide su4wulNinw of pancreatic aeetclion, Ihe level of total irnmunoreaclive elastase 2 i. p.ncre.tk renow plasma increued seven- to ten-fold, a.d the ratio of the iighter to the heavier mNerial increased eight- /oId The immunoreactive clauaw 2 in peripheral venous plasma is elevated two• Io live (old /4.114+winR srrmulaion, and an additional peak of proclastase is dectect.Me The kvets of immunoreactive elas/aae 2 in plasma samples from blood drawn Irom the mesenleric and pnrtal veins were essentially the s.mt as those in ohe perrpherat circulation ihese results suggest that pancreatic 12 ; procls.laje 2 eMers the venous hkwd directly from the pancreas and Is either rapidly cleared from the circulation or is bound Io.,-protease inhibitor. Gco4.s, M. C. rr at. Anrerlcan lorrnd of Physiology 218 (Gastrointesl. liver Physiol. 1):02)/{- - 0246, 19110. Otf,rr .urPort: Medical Research Service of the Veterans Aderini.tration. From the Enzymolopr Research 1.aloratory, Martinez Veterans Administra- tion Medical Center. Martinez. Cal.. and the DepartmeM of Medici.!• Uei' .enity of California School of Medicine. Davis. ANNOYANCE RESPONSE OF NONSMOKERS TO CIGARETTE SMOKE Nonsmokers rated their annoyance with the tsk environment while they were either e.posed or nd eapuscd to ambieet tobacco smoke during three dillerent eaperaoenls. The degree of .nnoyance was reported either with re/er- ence lo three kvels of noise intensity or while performing rnuhipticatiow prod kms under conditions of (1) high and «)~ ~a' ~~ ~ No diRerenc.M I smoke source was either a smoke pump in annoyance response to nuise was found as a/unctioa of smoke eaposure. Ilowever, both usk nwlitation and eaposure 1o ambient smoke influenced the nonsmokers' annoyance resporne to the task envbonrneM. Under low twoti- vati.g coaditions, nonsmokers eaposed to ambient smoke were more anooYed tlw those who were not eapoad. When 1he7 were highly rnotieated i. tM task ernrirorwnewt. however. noasewken were sipriAcsNNly less annoyed dum the ones who were nw eaposed to sswoke. Tht:ss resrMs waesl 1hat. N the very least. eaposure to cigarette smoke does aos increase annoyance with the environment whes nonsmokers are highly inrolved in the task. This tnay be worth considering iw the e.alustion of wrne tquests for smoking burs is workorientcd situations. Stoae.1. D., flreideebach. S. T. and Neinrsaa. N. W. PerceprMd and Motor StIMs 49:907-916, 1979. From the Department of rs)rcholoRf• University of South Dakota. VermiRie.. ECEr ORS IN HUMAN HAIOOCYf7C EUKOCYTB4 AND OPIATE TAia study deals PrinuriFy wilh weulrophilie~.rwlocresfl~ fm~ cytes, both of which demonstrate slereoqecHk opi Wnd~• o'wained healthy donors *as sutsrnitted to eounter/low tewtri/upNiot and the isdated monoeytts and kran'rlocylts wect carried on for /ursher Mudy. floth ceM types e.hibiled slereospeei/k. hi41r-dMnNy binding wish )-oluinuclidiayl ben:itate and dihydrornorpbist. apeeilk IiRands for the wws- earink choliner4ic asd opiate receptors, respedi.ely. Ttre slereosekctive wws- earink binding occurred in iM.et. viable celb as weR as in oep /raanc.a prepared by sonication. The apparent KA for the muscuink binding in 4raw- loCyks was /6 nM. and the number of binding sites per cep was om the ader 3l

"e of • X 10•; nanocytes contained approsimately I x 106 binding sites per ceR with an apparent Ka o1 20 nM. Studies is sonicated and in rarnnnicaled ceU pneparatios indicate Ihat the dfiaity of the 1-isomer is approaimalely 10 times that of the /-isomer. Soth cell types also eshibited Wereo.pecific opiate brndin6 with an apparent K, o/ 10 nM for panulocytn and I nM for monocyles. The .umher of binding sites per cell was 3.000 and 1,000, respectively. 11 seems that the observation of sler.oaeketivit7 in both cell receptors would lend support lo the Idea that the awscari.k binding sites have same (unc- lioeal capacily; the siRei6cance of the alenoacbctivit7, however, remains unclear. Lopker, A.. Ahood. L O., Hou, M., aad Lio.etd. F. 1. 1/ocAewrk.f IAarn.arology 29:1)61-1)63, 1900. Other w p p.r1 t U. S. Public /lcahh S.rvia. Fronm the Center for Brain Research, UniversM7 of Rochester Medical Cenler, Rocherer, N. Y.; and the Center for Rlood Resurch, Boston. ELECTROPHYSIOLOGICAI. RFSPONSFS TO ETHANOL, PENTO- SARR1fA1., AND NI('OTINF IN MK'E OENETICAI.I.Y SFLECTFn FOR DIFFERENTIAL SENSITIVITY TO ETHANOL Lon6 sleep (IS) and short Weep (SS) etice, selected for differences iw sleep lirne following a hypnotic dose of e/hand, were used to study cortical ekctroencephalnpaphic (FFC.) changes following injection with ethanoi (4.) and 1.4 g/kl), pentobarbnal (50 and 16 mg/kil), or nicotine (1.0 pg/kg). Ethanol (4 3 gfkg) caused EFO chanies in the LS mice that were quite dramatic and consistenl with a dhtiect slowing of the EEO with a con- comitant large Increase in amplitude. The SS mice showed a significant but ks% severe slowing of the f:F.O. No differences between the selected lines were observed following the activating dose of ethand (1A g/kg). These data sup- port the siotio. that the known difference is eth.nol akep time is due not to sreater SS sensitivity 1o ethanol aclivMion, but rather to greater 1S sensitivity to ethanol hypnosis. No diQerences between selected lines were observed fol- lowing 50 ws6/kR pentoharbilal, which again parallels previous behavioral data. Nicotiee nare severely reduced EEO power asd heat rate in IS mice; L eo.liMwrw iMravenous infusion o/ nieotine elicited a dirincl pattern of havioral Mer"py fot each selected Nnt, with more profound motor and rellea depreniow in 13 mice. Since these Rnes do .ol differ in rate of nico- tine nulabolism, they must differ in central nervous system sensitivity 1o nico- tine. Thom the lines of mice selectively bred for differential sensitivity to ethawol also display snarked differences in eleetrophysiological and behavioral tespodes b aicvline. Ryan, l.. l. a of. (AfcCfr.rw, G. E.) Jawwd of Co+np.arlve and rArdofopk.l hytAolopy 9)(6):101s-s0S2, 1979. Oth.r arpp.rfr National Instltule of Alcohol Abuse, and Alcoholism. From the Department of Psychology and Institute for Behavioral Geneties, University of ('olorado, Boulder. 54 I F.FFMTS OF CIIRONIC ADMINISTRATION OF TOBACCO SMOKE TO MICE: SEIIAVIORAI. AND METABOI.IC MEASURES This paper reports on the relationship betwee. activity in the presence of smoke and brain levels of nicotine. Itn this studr, male aad femak mice from four inbred Mrains and from two lines sekelively bred for high activity (HA) or low activity (LA) in an open field served as subjects. Tobacco anoke was administered to 1he test anintals during 14 dailp 10teiw. pretreatrnenl sessions in a boa filled with smoke frorw a nonRNered cipreNe. When ope.-fieht ae- 1ivily was subsequently measured in the absence or presence of antpke, pre- Irealed mice had signilkaaq Io..er activity scores than eorMroh. Comparisons of these scores under anake-present and snwke-abeM eonditiom revealed that the eAecta of acute esposure were depewdeN upow genotrpe. As tneasurad by open-Aeld activily, development of lokrawee 1o the effects of tobacco sanoke a/ser chronic prdreaheent was also 6enoqpe-dependent. In adAyion, si6niRcw pewoaypic diRcrences were found for wicotine remaining M Mvcr, braiw, ad blood samples when mice were sacri/lced 2.5 or i atin. after a weisM-specifk injection of radiolabeled nicotine. The tissue nicotine levels were also rela/ed to sea, tiree after in}eetion, and pretreatmest interactions with genotype. SpeciOcaNy, resuhs showed strong positive correlations between the n+easurea of brain and liver levels aC 3 nriw. after Injection and the behavioral measure of open-fkld activity under 12re smoke-present cond'Nioe. Raer, D. S. NcC/ea.w, G. EE and Wilsoe,l. R. psytAopArnr.rolofy 67:1)1-177, 1960. From the Institute for behaviord Oeeelics. Udvetsitp of Colorado, botddcr. TECHNIQUES FOR THE CHRONIC CANNULATION OF THE JUGULAR VEIN IN MICE The use of indwelling vesous eannulae in freely wwvity ran aed eNa hn become standard practice in the testing of drug ef(ec1a. Until ww, however, none of the canwulae available has been /ou.d suitabke for applying ahib lechnique to taice, the stadard teat animah for the preliminary analysia e/ drug effects. The ideal procedure sho.lA be SexibM erwugh to pttaait both repeated and one-time lesting awd be suAlciently, rapid and reliable to be applicable to large poupa of subjects. Tltis report describes the desij& of tw. eaeeulae for implantation iao the mouse jtrRular .eiw as well as the tech.ispin and equipment that have been developed for Ihe repeated intravenous adawiw- istrataw of drugs Into these awirwah. A ehtonic infusion chamber and both cnwwla designs are illusualed. Insertion depths for three different strains of mice and various body weiRhts, a well as estimates of operative wartdity are given. It is concluded that Mrplantatiow of cannulae of either desip Into the jugular vein of mice makes it possible to esplore many previouslr w- approachabk areas of eapetimenlation. Wwh eaperience, the eannulat caa be maintained iw place for at kasl two weeks, and the procedure is both rapid aad efleelive. Rart, 1. E. er d. (61rCfern, G. E) , PA.rnr.roloTy eiocArwdsey I A.Aa.io. 11:1/3-116, 1979. From the Department of Psychology. University of Cobrado, tloulder. J, 55

. GFNF.71('AI./.Y 1)1? f1iRMINFD SPONTANEOUS ANI) PEN 1 YLLNI? I- 1:1 RAIOL-INDUC"ED BRIEF SPINDLE EPISODES IN MI( •E In this search /or ekctroencephabgraphic (EFG) phenomena that rmghl be slrain-specifk, 1)RA/218(3 mrce, both normal and drvgged, displayed episodes of bilaterally symmelrical, high-amplNude 7/s spindles. lhe spindle, as noted in this paper, is an individual wave eornpka; a series of three of more coMinuwfs spindle unws without iMerreninll nonspindle activity is called a brie/ spindle episode (SSE). SSEs could be san to occur earlier in more (ronlal denvalions. 1 he SSIS appeared so actively suppress overt baly mnve- ment. though some facial twitching eodd be observed. Two convulsare adents, pentyknelelrarol and picrolotin, provoked IIISE activity; a third, nicaine, aup- pressed SSE activity at both sub- and wpr.eonvuls.nt doaes. lhree other in- bred nause slrains were also tested for SSE occurrence. lwo, ('3751./6 and SA1.1I/e mice. failed to show 1lSE activity either spontaneously or aller pen/ykneleurozol. 7he thud sl.aiw. C)1I, showed lower•nnplitude, shorler. dwalion and less wepor6ani:d RSE activity. luvenik DeA mice showed slo..er, lower"amplilude, shorlerdwMion 1SEa than did aduhs. Older mice showed more aduh dSlh. The fesuMs here raise the possibility that 9S1?s are an epileptic phcnomcwon, and this is discussed. 11 is ckar, at kast, that they are agencyicaMy determined ekcrroeacephalographac characteristic and that the DSA mice may provide a use/ui animd model for studying Ihe genetic basis of the inheriranct of an epdepu/orm phenomenon. Ryan. (. 1 and Sharpku, S K (MrCle..w, G. E) Experlmenrd Nr.urniory 66 19)•SOe, 1979. From the Ikpinmem of Psychology. UeiversMy of Colorado. doukkr. EFFECT OF NICOTINE ON OVIDUCTAI. I.ACTATE DEHYDROGENASE DURING EARLY PREGNANCY IN TNE RAT I• this attempt to ehrcidale the control mechanisre responsible for a den+onstraled delay in ovum cka.age, attention was focused on the lactate dehydrogenase (LINI) of the eviducts of rats /realed with nicotine during the preimplantation period. For this Nudy, rats were injeeled s.e. with either nicoliwe (7.1 mg nicotise tartrsle) iw 0.2 rwl aaline or with saline (0.2 ml) twice daily from the morning of proestrw (Day 0 of pregnancy) until the day of sacrifice. Rau were sacrificed be/ween 1000 hr and 1200 hr on 1)ay 2 ar Day ) of preitnancy, o.iducal Bushings were eolketed, e.iducM were IsMnrwg- reited, and the Bushings and hornornaks were analyzed ekcarophorelrctlly aad apeetropholometrically. FlusMnp of .ico/ine-treated rats showed a sit- siAcanl i.crease in specific activity o( 1.DN on bolh Day 2 and Day ). There was wo aheration in the distnbWiow of any of the Iso:yarcs or in the percen- 1alt of S sub units in the 1)ay 2 oviductal honwgenates of the nicatine tretled rats eompared with the conlrols. Noweva, in the 1)ay ) sampks, the con• centration of I DII S relative to the total was significantly kss in the esperi- menlal group, and the percentage of S subunits was significantly greater. : I Nianine caused marked aherations in Ihe relative concenlrations of LDN isorymcs in the oviductal Ilu.hinp of both groups of pre6narN ra1s. Overa/1. Ihesc results suggest thaf, unless l.q/l eaer/s an iehibrlory eAecl on develop- meM at the high concentrations found in the oviduct of the nicotine-Ircaled rats, an add"aional mediator must be implicated iw the regulatory (unclion of nicotine on the development of the fa/ osrfot. Rice. C. and YosAin.g., K. Sidogy o/ Rep.oducrion 27:113-451, 191/0. From Ihe Departments of Anatomy and Physiology. Harvard Medical Schod, Sosloa. V. Pherwfero/osy and eioctiefwi.try I THE ANTI -PROT EOLYI IC BEHAVIOR OF LATHYROGENS This study e.anines the possihility that the lalhyrogens, p"arninopropiowi- tnk and .-aminoacetonilrik, eaerl an e/leet on praeulytic enfyrnes. Results reported c.rlier have shown that S-aminopropionilrik primarily inhibits lysyl osidasc, a key entynse in the bxmation of both eollagen anJ elastin crosslinka, by catalyzing the oaiddive dcaminaUun of eertain lysyl residues in the two irnpwtant structural proteins. Rewlts presented here denanslrate that bolh lathyrogens possess anliprofeulylic activity against trypsin-like entymes. At concentrations of 100 mM both cawpmads are very eAecuve inhibitors, where- as at low concentrations only little e/lect or s1i& slinwlalion was observed. l.inewtaver-Surk plots show that Ihe iahibilinto is competilive, with .-afniwoacs- hrnilrrk being the more potent inhihil.x. Ahw, the enzyme associated with, and capable of digesling. Irnpoelas/in is inhibited by both lathyro6en+ wheo tested against its nalural subslrale, Iropoelastin. In another aspect of this uudy, administration d.-sminoacetonilrik•11('1 to the dief of young chicks resulted its a 62% incrtase in the yield of Iropoelas/in and significant reduclioe in fatality as compared to p•aminuprupw~nifrik-lumarale. "Ihis 1sl lalhryoges has recently gained usage in the study of connective tissue praeins. Its ability Io inhibit proteolytic enzymes is an Important properly to realiee in the desi6s and assessmenl ot biosynthnic and dc6radati.e eaperimenls where synthesis and turnover are being e.amined. . Foster. 1. A. er et. Siorhiniirn ef R1ophyiiru Aria 5117.d77-JS3, 1979. OtAer .urr..rt s National Institutes of Health. From the Ikparlmenl td Diochemistry. University of Georgia. Athens; and Ihe Ikpartment of MedKine, Washington Uuiver,Ny School of Medicine, St. Louis. 37 56 0

i i TRANSLATION OF CHICK AORTIC El ASTIN MESSENGI'R R1B()NU('LEIC ACID. ('OMPARISON 10 ELASIIN SYN1111:SIS IN CIIICK AORTA ORGAN CULTURE In this attempt to define the molecular size of the elastin prirnary gene product, cell free translation of aortk elastin messenger ribomrkic acid (mRNA) Is described and the Iranslation products are compared to those synthesized in anrtic organ culture. 1n these dudies, translation of chick .ortic mRNA in an mRNAdependeM reticssloeyle ITsak resulted in the synthesis of two major prdeins of 70,000 and 7),000 molecular weights. Bm3 proteins were shown to be soluble forms of eiastie by isotope incorporalinn. immuno- p.ecipitation, cdlagenase and cyanogew brorwWe sensitivity. and Iwo-dimen- sional gel ekctropho.esis. The 70,000•dahon protein behaved siwnilarly, to authewtic tropnelaain in sodiuw dodecyl sulfase pcl e/eetrophoresis. There was so evidence for a high molecular wcidM /onn of soluble elastin. although procollagen chains were indirectly identiAed awtong the sottk rnRNA-direcled translation products The same wrokcv/ar size proteins were also sees in organ cultures of chick embryonic aortr labeled with (sH)valine. Signili:antly, the 7),000dahon protein was not estractable i. a neutral siih buffer but was found only if the aorsas were Irealed with area in the presence of refucinll and alkylatinT readents. The results from these studies suggest that ela•ain is Brst synthesized as two distinct polypeptide chains that differ slightly is size and overall charge lhe pnssitulsty that these two proteins may associate post- transtatsnnally to lorm a drmcr prior to secretion is postulated to e.plain the esistencc of a putative proelatton molecuk seen in dher systems. Fosrer, I A er el d'aefie.nlrrrT 19(S).kS7 e61, 1960 Other asr/ r..rr: National Instnuus of Hca11h and the National Foundation- March of Dimes. From the Department of Biochemistry. University of (ieordis, Athens A SENSITIVE ASSAY FOR THE QUANTITATION OF SOLUBLE ELASTIN IsKrea.1ng interest in the biosynthesis and degradation of elastin has made the .vailability of a rapid lechnirtua for quantitating soluble e/.slin in tiswe tstr.cts awd /w rNre systems dcskable a.d signi/kant. Currently ealstinR Iech- stitlues, while sensitive and applicable to crude estr.cts, either require large quantities of .misera or usually Involve ss'1 labeling of .nften. TTus eon.- nwnication describes the development of a rocket imnwnocleclropbaetie system that can be used for the direct quantitation of unlabeled and end- ogewouslT radiotsbekd sohrbie elastin in crude tissue estracts. The techniaue which combines the idemifkation criteria of both ekctrophorNic mobility and imnrunmeactivNr, is applicable to samples of unlabeled /ropoclnlin in eow- centrsrrorss ranging lrnm /O to 100 mo/l with the aid of direct ('oomassie blue surnink br re.wJ .enuuvq7 (1 mo/l) was obtained through the use of in .wu rsJrdaM/kd u..pKFavm and of fluoroTraphy to dclecl (rH)valine- sa t 1 • . I labeled tockets Immunoprecipitalion produces a ma}or (s11)valine protein with a mokcular weight of about 70,000 and electrophorelic mobility identkd to Iha1 0l purifkd N.B(sH),-labekd tropoclastin. lhe rocket immunoekclro- phoretic system was recently applied to the cell-/ree trandatron of dsli» mRNA and can now detect 0.05 pnwl levels of elaslin. Its sensitivity ahouid make the rocket technique an eaeelleal tool for studying elasliw synthesis. Forh., 1. A. et d. Andrrkr aiocbenrlirry Io1:710-)13, 19t0. Other ar'Fertf National Institulea of Health. From the Depanmeel of Biocheminry. University of (ieorBia, Athens. SUBSTRATE SPECIFICITY OF HUMAN 1'ANCREATIC ELASTASE 2 As pan of a luker atudy .imed a mwe clearly de/hNnR the aalwe ee elastolysis and /urther characterizing pancreMie elatase 2. this report esawd.es the enzyme: speci/kNy. ranicular emphasis is placed on the peplide bond specilicity, which is compared with those of the other characterized elastasa and bovine chymo/rypsin. A series of peptide lo-witroanilides were used as substrates. According to the resuNs, humaw pancreatic elaslase 2 appears b have a peptide bond speci/icity similar to thN of porcine pancreatic elastae 2 although it is signi/kantly diAerewt from that of porcine clawase 1. The evi- dence suggests ahm the enzyme also has an ealesided binding site whkh imeracts with at least live residues of peptide subatrNes. With the oompowds tnudied, the overaM catalytic e/Rciency of human pancreatic elastase 2 is signi/icantly lower than that of tither porcine elslase I or boviae chyrnwrypsi:. Tlrcse data support a previous st.yestion thN NassiAc.liow of elastases as a separate fatnily of seriee endopeptidsses-as deAwed by a certain pepide bond speciflcily (preferentiafly hydrolyting .1 Ala residues) and the abilily • so hydrolyze elsstin-is no loneer adequate. Continued grouping of elaslases .c- eordinR to elastolytic activity is uu/ul, however, because of the physiologic Implications. Del Mar, E. O., L.rpeaw, C.. Brodrick, 1. W.. Fassett, M., and Geni.r, Af. C. IiotArwrbtrr 19:16t-172, 1980. From the Enzymology Research Laboratory. Martinez Veterans AdmMhtrMio. Medical ('enter, Maninez, Cal.; wsd the Departnsent of Medicine. University of ('alrfornia School of Medicine, Davis. . A SENSITIVE NEW SUBSIRATE FOR CIIYMOTRYPSIN Evee though chymotrypsin is an importuN snd widely studied endopep- tidase, no convenient and sensitive substrate for its determinatioe had ea- ated before this time. Now, in the biochemical study reported here, a.ew 59

. I i sulnlrale lur chynkrtrypsrn. succmyl-Ala-Ala Pro-Phe-p nitraamlKk, was symhesrteJ and characlcrrtcJ Krnctk rn.dysis of the hydrnly%rs of this pep- tM/e by bovine chynwrrypsrn gave • K. of 0041 mM anJ a A„$ ad 45 s- r. The new suhstrate, a white s.An/ wNh a uv ahsrxptmn matumum at 315 nm, is readily soluble .nJ stahle to hydrolysis in 1 ris hu/ler at pI l M. ('hymo- uypsrn levels down to 1.0 nll can be rapidly determined spectrophotumetrically. IA•1 Mar, 1=. (i . I argman, C.. Srodrick, 1. N, and (%ru4as, U. C. Analyritu/ si..rl+rrnirny 99:116J20, 1919. OtA.r •urPorz: Medical Research Service of the Veterans Adminislration. 1 rom the 1-ntymnlogy Research l.aboralory, Martinez Veterans /Wmioi.tration MeJ.cal ('enter, Martinet, Cat. 111F MF1AS(N IC N OXIDATION OF )-SUSSTITUl[':D PYRIDINFS IN VARIOUS ANIMAI. SPECIES IN YfYO ihn systematic study of heteroaromatic N osidation u.e+ a series of l-suMtNwed pyrd.nes as maxlel compounds so as to eslahlish the enrynx.bgy, oof the processes invdved Results of previous in vitro studies show that a general metaMl.c pathway for several l-substitnted pyridines seems to he o.idalia.n at the pyrdyl nitrogen. In order to determine the rok of N-osida- lion in the metabolram of Ihese compounds, the in viro metahulism of pyri- dine anJ of its I chbro anrl 3 methyl derivatives was ree•amined. Intraperi- tonea) admrmstratn.n of pyridine, l-mcthytpyrdine and l-chluropyridine lo mice, hamsters. grnraa p.p. rabhits and ferrets produced urinarN e.cretinn of /heir N osdes In all the species uudreJ, pyriJtne-N-oside was a quantitatively siRm/kant mctahulrte ul pyridine, the percentaRe of the oriRina/ atenl's dnse eaereted as the oaidc ranging from 10% In rats to ahout 40% in mice and guinea-pigs. Other osiJn. )-chloropyridine-Noaide and l-methylpyridine-N- oside, accounted /ot less than 7% of the administered dane of the parent compound. Urinary escrNion of pyridine-N-onide was appreciably greater in mice pretreated with phenoharbitone lhan In control mice, but pretrealment with i-methykholsnthrene did not have any nolabk e/lec1. (:o.roJ. /. W. and f)amani, 1.. A. t:rroptan lournal of Drrt Mt/aldi.nr anl PAarmaroklnrrks S(1) :Sl-S7, 19110. 1'rom C'helsea College. University of l.ondon, l arlon. 71/F IN Yl1R(1 MFTARt)/ ISM OF l-A(-fTYl PYR11)INF.. 1 At'1 IY/ PYR11)IN1 N()XIIW, ANI) I-(-) PYRII)Y1.)-Fi11ANO1. whrrr.. I ...~slp~nJ..w .s a a.d..uar /ur .yuKhrnmr mtduteJ N nRr. .1.,,.,,. t p-1.4..r...J .. .rr Iw ..u.. .n.tau.Ralw.n n/ the mtlafr.lrc route I..n.•+.J I.r I . r..t/ .~.1.... I 11 pt-Atl/ roh.n,d 1.atlylpynd~nt N otele. s i 1 1 1 anJ 1•(1 pyriJine-N-uaiJe)-elhanad showed /hat hepatic microsomal syslem~ cnnvert the first compwnd to )-acetylpyridine-N-oside, while a soluble hepatic enzyme reacts with i/ to form 1-(.I.pyriJyl)-ethanol. The flrst product is readdy redrKed to 1-(•l-PyrMyl-N-osiJe)•elhanol and the second reaisls Not- idatiun. 7hese resul/s indicate that the mainr metaholite of l-acelylpyridine is formed as a result of heterocyclic arnmalk-N-o•iJation fol{owed by c•r- honyl reduction. In addition, hecarseal'ight oaidation of 1-(3-pyridyl)<thanol, which was enhanced in Ihe presence of NAD, was aho noted with ankro- aomal fr•cliuns, it is suggested that lhis niicrosornal carbinol dehydrogenase might he the same as that described by ahen for the dehydroRenalion of irdanol. GorroA, l. W. t1 al. In: Coon, M. /, rr .l. (eds.): Mkrosorrwz, ArrR Os/daHons and CAtnskJ C'arrinorrnrsis, New York: Academic Press, 1980. vol. 11, pp. 961-161. From the I)eparlments of Pharmacy, Chelsta Colkgt, UnivenNy of Londoa, l,ondon; and the University of Manchester, Manchnter, England. TIIE ORIGIN OF 1-( l-PYRIDYI: N•OXIDE)ETIIANOI- AS A METASOI.ITE OF l-ACETYI.PYRIDINE In this biochemical sludy, an atlempl was made (o understand 1he aui- ous faclors involved in heteroaromatie Nolidatiom by studying the wtets. holism of l-acetylpyridine, l-acetylpyridine-N-oaide, and 1-(l-pyridyl)e(Aaad ln vino. Thir-layer c\romatoRraphy, ps.liquid chroma/oRraphy and mass spectrometry were earried out uw hepalie tissue fraetions from wsak ra1s, fuinea-piRs, white rabbits, hamslen, utd mice. Following wilabk iwcubNiow 1 rocedures, results of these studies showed Ihal :(1) 1-atcelylpyridMe was mctabolized eaterni.ely to /-(l.pyridyl)cUuwol whew /he hepatic .vurce was the twhrbk wpernnaut; ).ace(ylpyrdine-N.oaide was identiAed as a nKta- bolite by usinR either the IO,0t>OR or Nre wricronom.l /raetion; (2) 1-(l- pyridyl-N-oairk)ethanol was wol deteeted as a melabolile of /-131-prrldlrl) ethanol using tissue prepara(ioes. Hosrever, 3-acetylpyrid'ase was formed ia trace aemunts when the alcohol was incubaed with either the 10,000R or /he mkrosomsl fraction: (3) IncwbNioas of l•acelylpyridine-N-oairk with the snlubk or 10,000R (racliun reswdted in the forwation of 1-(7-prridyl-N-oslde)- ethand (keto-reductMn) as the rnaiom melabolile. )-Acetylpyridlne was formed in trace amamts (N oside reduction) with bn/h the 10,(/II0R and she mkro- somal Iraclions. O,rerall, therefore, i1 appears that the rorqe to 1-(7-pyridyl- N•oaide)ethanol, recognized as a piincipal urinary metaholite of acetylpyridine, is via N-osidtion of l-acetylpyridme fol/owed by earbonyl reduction. Damani, L. A., 8ryan, I. Cowan, 1). A, and Go.rol, l. W. XrnoAioNr. 10(7/e):615 651, 1VR0. From the /kpartmra of Pharmacy. Univenity ol Manchester. Manchesler, I:nC1anJ; and the 11\•partmcnt of Pharmacy. Chelsea C'olkpe, University of I onJun, I aunJun fit

I I EFFfiCTS OF CIGARETTE SMOKE INHALATION ON THE CAT EI.E(TRORETIN(X3RAM. COMPARISONS WITII NICOIINE AND CARBON MONOXIDE Carbon monoside (CO) inhalation has been shown in the past to resuN in a signilicanl ekctrorelinopaphic b-wave depression in anesthetized cats. The nudy presented here was done b delerrnine whether cigarette smole inhalarion under similar condiuons wouW a!w result in a b-wave depression. f•aralkl audies with nicotine and taicotine-CO combinations were done, too, so suess their rok in any possibk snwbe effects on the dectrorelino6r•m (F.RO). In the Arat studrcs, can Inhaled the srnote diluted to varying de- gree" with air for one minute every 30 wriwules over a 1wo how e.perimental period. lhc comparative studies were done with i... nicotine inicctions and infusions (5-100 rl/1.6) and with sinwNaneous ivI nicotine and CO inhala- tinn (0(IS 0 196 In air). Artcrial blood presswe, heart r•te, userial oayhemo- glohin. carhosyhemotlobin (fIbCO). pO. and pCOr were routinely nwni- /ored alonl with the dark-adapted ekelrotetinographic response to Aashes of whnc light Data show that cigarette r.oile iahdation rewMs in a si6ndicant dectemenl in mean b-wave amplitude (ristaaimwn 16lL ), roughly paralleling efterial IIhCO (masieMUn •N11. IMraveeas nicotine alone had no si6niA- Larw effect on the b-wave amplitude and when combined with simuNaneous CO inhalalion, nicotine did not silec/ the ability of CO to decrease the b-wave. These resuhs reveal effects of cigarette smoke on the pretanglion cell relina that are similar 1o those ol CO. Ing.niro. A. /. The /ournal of rh..w..rolor7 and EzKrbw.wrd TAer.yeMrks 211()):667- 635, 1979. From the Department of Marmacology, The Albany Medical College of Union U•iversily, Mb•ny, N. Y. EFFECiS OF CARBON MONOXIDE ON THE'E b-WAVE OF THE CAT F.LECTRORET1NOaRAM: COMf ARLSONS WITH NITROGEN f1Yf OXIA, EFINErNRINE!, VASODILATOR DRUGS AND Cf1ANQES IN RESPIRATORY TIDAL VOLUME Iw order to dNermine it low levels of carbon nanoside ((b) dfea visual turrciiow by acting •1 the level of the prepn6lion cell retina, dart- •dapted ekcirorelinograms (F.ROs) of ehloralose-awesthetiud cats were nani- Iored durinR • two-hour eaposure te 0.03 a+d to 0.1% CO in .ir. Resuks were eanpared to those obtained with the following groups ot cats: (1) con- trols eaporcd to air alone. (2) thoae eaposed to a witroolew-air m,ature, (3) those with chanjes iw respiratory tidal vohrne, (4) those having vasopressor respornes induced by epinephrine, and (5) those with easodepressor re.potaes caused by vasodilaors. The last three intervewtioni were iMe•ded to simulate swee of the cardiovascular and respiratory eAects of CO and Nr. Carbon mnnosKfk' decreawd the FR('G b•wave amplitude as the arterial carhoarhemo- tlabrn (11h('t)) rncreased, with a si6niAc.n1 me•• decrement (10% ) oeera- rrng in the "environmemai or smokrng" I/6CO range (7.396 ). Nitroge•- 62 . indnced hrpn7he aiad. on the other hand. it anythin~, tended to nrgrnent the b-wave arnpliwdes au presented here suggest that the laltcr effect Is due wui•y 10 hypocapnia secondary to N. induced hrperpnea- The release of e/ri•ephrl•s during CO or Nr hypoaa Iwobaby did no1 signipcaMy inM.ence their ERG effects since intravenow injection of e ~aiAtaotly dletl the h-wave. Several ot these observations w~~ine did .ot .i b-wave decrement may have been caused brdireey to:ic effect on 11re r rather than by IIbCO-mediued retinal hypowia secondary 60 a decrease iw ar- terial osyhemo6lohi•. Only minimal CO eQecls were awted os other par•- meten of the dart-adaped cat ERG. In`rnito, A. /. and Durlacher, L. TAe /ou.w.l o/ rA.rm.rolotr and EzperJwkwrd T4r.prrrkz 211( l):6)11- 616, 1979. From the [kpartmeM of Pbarmacolooly. The Albany Medical College of U•io. University, Alb.ny, N. Y. I FACTORS AFFECTING THEOPf1YLLINE CLEARANCES: AGE. TOBACCO. MARIIUANA, CIRRHOSIS, CONGESTIVE HEART FAILURE. OiESITY, ORAL CONTRACEPTIVES, IlENtOD1AZEf+INES, BARBITURATES. AND E7HANOL While dosage reauiremenb cat be predicted teasonahy suoceafuMy, tor drugs whose ckarance is largely deternrined by "a.l tuwction, as tueaswmd by serum cre•tinine concentratiow or erealiniee ekarsstee, twmerow tacun 1hM aRect biara•s/orrw•tion rates wtoke dos.Aen bighly, unpredicyable for en- knsivcly metabolized drttAs. One ot /htse. 1Aeeplaylliee, poses clinical pli.r. macotinetic diA{cuhies. CowuqteMy, it te iwrportant to develop pideli•e. for its ts.e in diAeresN types ot patients when1 y~rrrati~ trotw theepbyNiar assars is wot immediately avaiiabk. TUit repwt deserihes eanruoe deterwd.rl t•cton of total body /heopAyMi•e ckar.nce (C7.). f Aarmacotinelk n•d sl•- listkal methods were wed to eh•e•cterist NrcopAyMine clearance variability i. 200 srbjects iwchtAiwd aduN pWr•ow.ry ligilietwo. pe"ric plfeuw .orad adult vdtnween, and otAers witA pnicsrl•r problems twcA as obesity •.d cirrhods. The major tac/on affecting C/, itt this population were ye, liver disease, amoking seNw and congestive heart failure. Age correl•1ed strongly wilh C/,. Specific wrbAroups produeed additional variability. Young aduN (20- 10 rears o( age) tobacco/wt•ri4u•n• user. had tht hiRhest CI, values, whik older (over 10) patkMS with hver disease 1md the lowest. Iw certaiw wbject cNegories, sea (in kewagers), oral eowlracepNves (in srttoters), obe.ity (i• young nonsmo/cen). and barbitwatq aho affected theophyNine ekaranoe. O0 the other hand, several common (aclors such as chronic theophylline or cor- tieosleroid usage. caReine, sea in aduNs), and pregnancy did not aNer it. The trriaue approach to drug clearance characlerkaliow in paienla presented here identifies criteria In a manner that may be useful tar predicting drug ckaranee rates and dosage reilimens. /rsho, w. /. .r r. /onrnd of rAur.nor.urkd Scienrcr 6t(11) :1)S8 -1)66, 1979. 63

Otli.r .upporl: National Institutes of General Medical Scieoces. From the Clinical Pharmacokinctics Iaboratory, Department of Pharmaceu- tics and Medreine. Schools of Pharmacy and Medi.me, StNe University of New Yort at Suflalo, and Millard Fillmore Hospital. Bulfalo. THF INCORPORATION AND I OCALIZATION OF AI.DFHYDFS • (111(;HI.Y REACTIVE CKiARE17E SMOKE COMPONENTS) INTO ('Eli ULAR FRACTIONS OF CUI-TURED HUMAN LUN(i CELIS The resuMs of incubatinN W1 )• huwun fetat IunN AbroMasts with r'C- labeted formakkhyde and ucetaldehyde uwder contrdkd conditions are de- sarrhed here In one s/udy, the miFration of formaldehyde from supernalaM into cetl nucleus was (ollowed with pulse<hase eaperimenls. Readings at 0 min. MI min, and 24 hr. showed that the 1o1d amouM of counts per minute in aN IrKlans remained essenlraN7 the same (or each period, but the dis- rrrbuti.wt of coums changed with time. Wi1h increasing lirne, counts from both the supernatant and memb.ane fractions decluted and appeared in the nuclear portion to anaher series of e.periments, fractionation of the nucleus revealed that the RNA tractan had the hiNhest absohne and speci/k activity while the DNA and protein fractions coalained considerably lower activities. When the nuclear RNA and DNA fractiona from the formaldehyde incubatiom were separated by Dowes Sto chromatography, Iwo peals (adenine and gua- nine) were found in the RNA hydroly.ale. while three peaks (adenine, gua- nine, and thymine) were apparent in the DNA hrdrolysate. On the other hand• incubation with "C' accla{dehyde showed a majority of coants localited in the membrane phn nuclear lipid fraction. Only a small proportion of counts was found in the protein portiow of the cells. Pruett, 1. 1., Scheumtuht. l/., AfirAarN. D.. and Nevo. Z. ArrAires of Ewrironmenrd Ne.IrA 7S(1) • 13-20, 191t0. From the DepartmeM of Siochemiatry and fliophrsies. University of Califor- nia. San Francisco; and the Department of Chemical PathotoLy• Tel Aviv University Medical School. Tel Aviv, Israel. N1COl INE REDUCFS EMeRYO GROWTH. DELAYS IMPLANTATION. AND RETARDS PARTURITION IN RATS • Since nicotine has been shown to modify Ihe process of implantation in rals, this study was undertaken so determine whether the naed implantation delay was assruiated with altered embryo developmesM. For this reason. the time of rona pctlucMfa loss, growth of the inner cell mass and rne of cell i . prolilrralNrn were delermincd in embryos retrieved from conarol and nicotine- Ireatcd rats The ellecls to nicotine on parturition and fecundity were also no/ed. Results o/ these esperimenls showed that daily injections of nicotine during the initial 6ve days of pregnancy reduced embryo growth, delayed implanlalion and retarded the onset of parturition. On Ihe other hand, nico- tine administration did not aher INter size, birth weisht, sea dislributio., ar mortality rates. While injection of pharmacological doses of niootine into /emak rats is clearly not equivakrN to iehalalion of the alkaloid by womea smoking cigarettes (di/ferences iw speeies, route of administration and dosage preclude eatrapolalion), the fact that nicoline eam nadify embryo develop- ment without being embryoloaie or impairing fecundity may render it tnelul /or studying embryo-uterine interactions. Ilarnmer, R. E. and MitcAelf, d. A. rrorrrdinps of rAe Sorirty /or Eaprrimrwaf e/otoPy and A{edkine 162:)I)- ) )6, 1979. From the Department of Analomy, Wayne Sta1e University School of Medi- cine, Detroit. EFFECTS OF THIOLS, SUGARS, AND PROTEINS ON NITRIC OXIDE ACTIVATION OF GUANYLATE CYCLASE Several osidation-redudiow processes activalr guanylate cyd.a• AIL though Ihe precise mechanism remains unltaown, one pathway eoswrwo. M rnany of she activating agents may be the /oraatioa of witric olide. Solrb4 rat liver guanylate crclae puriAed abow 9,000-(old is activated by nitric oada as weN as by nilroprrsside, hydroayl radieal, uro.luraed /aty acids„ and stable prostaglandin endoperoside analogs. Pattid purification of Me estsrawt e.u eause an apparent loss of re+ponsiveoeas 1o aiuie oaide and other aeto• pounds, which is restored by the additiow of haee proleies and other waN- riah. This report sugests that /hia app.retN loas of response is due 1o chwtila in Ihe dose-response curve, which att rerersi0le wNh dNhiwlueNd, boviM serum albumin, mnhetnoglobiw. or wcros.. The apparent loss of ewtymt respomi.eneas to hittKr wMrie oaide eoecemralions during pwilkation, there- fore, is probably a functiow of the eneynte's sensitivitT to oaidation, the bi- pha.ic oature of the dote-respoese curve, and the removal iw the procer of purification of nitric oaide scavet.Nen or sinhs, such as hen.e. proteins a.d small mokcuks, which prevent eaeessive oaidation of the enayme. SrauNhkr, 1. M., Mitlal, C. K. and Mural, F. TAe7orrnd ol slolotkd Cliemirrrr 2s4(21) :12430-12.5., 1979. Ofbtr support: National Instituta of Health and the Virginia litean A.aod- atioe. From Ihe Division of Clinical PharmacoloNy, Departments of Pharmacology asd Medicine. l)niversity of Virginia. Charloltesvilk. 65 64

O(('URRI N('E OF METHIONINE ENKEPHALIN IN IIUMAN PLACFNI'Al. VII.I.US The known propenies of enlephali.a and endorphitr suggest 'hal they may act as neuromodulators to regulate either the neurotraasmNler or actual horreone release through positive or negative feedback rystenss. Specidcally, entephalins may regulate neuronal release of acelykholine (ACh) and swr- epinephrine by .eptive leedhack systents. However, although it is thus con- ceivable that enkephslis might regulate placental ACh, neither the o:currence nor the role of c.tephalias had her.lefore reen demonMrated in M man pla- centa. The investi6atio.s reported hera i.dieale thal hanan plaeentd villw does contain biologicaMy active methionine entepha6n and /Jsmkephi., as determined by highly sensitiw awd qeeiAe radioinrnwso.ssaya. The actual eontewl varied fronr oacetMa to plaoeNa, awd ti>e diMributiow of winhio.ine enkephabn paralleled /hat of ACw. The biolosic activity of the placentd pepltdes on the intranwrally slinwl.ted rN and rwouse vas deferens and srinea pig longitudinal ilcal muscle also demoostraKd The presence of o/:oid pep' tides. It is, therefore. suggested that both r.elhioa.e entephalia and if•endor- phin may indeed eaen negative feedback control on the placeaal release of acetykholine, or alter.atively, that their placental release may regulate sea- aory IraMmiaion (or pain impulses) to the central nervous system ftorn The uterus and vaginal tract during puturitioa. Sasny, •. V. R. rr d. Siorbemk.l PArmarololr 29:175-47l, 1990. OOther support: U. S Public Hedth Suvice. Fran the I?eparttnem of Pharmacobp, Vanderbilt University School of Medi- eioe, Nashvdk, Tenn. 7OXIC.Ol.OG1('AI. AND PIIARMACOLOGICAL EVALUATION OF IIAI.(XIENATED CHOLINE ETIIERS fo this attempt to eWeidate the loakologkal and pharmacotojkal prop- erties of a serin ot choline eshers, The following parameters were considered: (/) scuta 1onkHies of the ebollne ethers and their modulation by spccUk choNnerRic seurs.irs: (2) the activilies of the compounds at various choll- nerifk siles (nsuscarinic receplors, .ieotinie receptors, chotinesterase. and eholine aextypransferaae). aad (3) 6e relationship of the chemical structure with the bsicob6ical and pharmacolo6kd activities of choline ethers. The ethen studied here were O<tbykho8ne (ethykholiae ether, Et('h, O•(2- chloroelhrl)cholase 0-(2-brornoethyl)cholise (erEtCh). O•(2-iodo- ethpl)choline (Ili('h), and O•(2•hldroayethrl)choline (OIIEt('h)• llrc loa- icities of these agents In male Swiss mice decreased in the following order: IF.t('h >erEtCh = CIFtCh >EtCh »OHEK'h. Oased on the toaicological properties eahibited by these choline ethen, three conchnions could be reached: (I) the onhaMpenated patent cornpnund (Et('h) was the most potenl mus- carinic ajtent in the series; (2) the halogenocAoline ethers ((YNN'h. Srl;t('h, and IFK'h). having similar aclivtty, were weaker muscarinic agents tham I 1('h in the serres, and (1) since the respiratory depression caused by chuline I a ethers other than IaCh at l.D_ dows was siRrrifkantly lowered by atropine pretreatment but no1 abolished cornplelely, it seemed apparent that these elhen produce their toaic enens by mechanisms other than stimulation of the ews- carinie recepors, which a.e mainly involved iw the maintenance of reapira- Iion. The ellecu of the choline ethers on the emacarinic and nieolinic recep- tors of The guinea pig 1ongiludin.t muscle were also studied to understand th- relationship of the cholinergic system to the loaicity of 14se elhera. Chaturvedi, A. K., Rao, N. (3. S., and S.ory,1. V. R. ToskolopP.wd Appllyd P6...nrolopp Sd:26S-27S, 1980. Other ar' portr U. S. Public HeaNh Serviee, American Caneer Society (North Dakota Division). From the Stte Toaicobgy /.ahors/ory, aand Departmem of Pharmaceutical Sciences, North Dakota State University College of Phsrmacy. Fargo; a.d /he Ikpartment of Ph.rmacobp, Vardabill University School of Medicine, Nashvilk, Tenn. ADENOSINE S'-TRIPHOSPIIATF-CITRATF. LYASE ACTIVITY IN MAMMALMN SPERMATOZOA. A NEW RADIOMF.TRIC METHOD AND CHARACTERIZATION OF TIII: ENZYME IN SPERMATOZOA Mammalian spermalo:oa contain choline acelyltrsnaferase (CAA), which ~ cataly:es the synthesis of acetykholine (ACh) from its precurson, cbli.e and acetykoen:yme A f ACoAI; but nahinS Is known about ACoA in sperm, nor about their content of ATPtitrale lyase, which catalyzes its synthesia. Comcquently, a radiomctric method reported here has been developed for the assay and characterization of ATPsilrale 1)rase iw rnamrnalian aperm.- tozos. This method is rapid. veryr sensitive and measures the formMiow of ACoA directly. 11 is also adapt..sle to the measurement of suboeNular frac- lions and enzyme preparalions separated by suerose density gradieMs. Sper- malozoa do uo1 have mend+rsne stores of ACh, indicating that its syrMhe.is, receptor stimulation capacity and hydrolysis by ACAE are closely linked .nd nmay be localized In the ssne compartmenl. Strch uniqun orpnirNion woai/1 permN eontiwuiw.s rapid A('h turnover. However, lu ptrcurson tnuH he read- ily availabk, and whde the sperm cell provides most of them. ACoA wwM be supplied continuously. That ATPtilrNe lyase is present in spermato:oa, as demonstraled hcre, indicates that ACoA can he formed from citrate. 7Le occurrence of Al Ptitrate lya.e in human placem has also been demon- straled by this method. lheref.we. A('oA can be formed from citrate in human placenta Chawrvedi, A. K. and Sae.y, s. V. R. IiorA.Mrkal PAarnurrof..cy 29:25119•2391. 1980. Othrr.appertr U S Public Ile.lthSetvice. From the Department of Pharmacok.ly, Vanderbilt University School of Medi- eine. Nashvilk, lenn. 67 66

FFFEC 1 S OF PF1O IOSENSI i IVE CHEMIC'ALS ON MALARIA PARASIIE (-ELLS A sensilive, rapid and economical in vitro assay procedure, which is based on the Incorporation of radioactively labeled adenosine into a macro- mukcular fraction of infected red blood txlb. is described here. lhe in vitro assay is then coupled with photoinactivation by 1-fluoro•)-nitrophenyl aride (FNPA) and NAP•nornicotine to provide evidence for a direct correlation between a light induced loss of infectivity in rlro and a decrease in the in- corporation of labeled adenosine in sdrr.. Eaperimental resulrs show that these two prohes can inhibit sqes that are essential to the survival of the rodent malarial parasires, r. bere/lrl awd r. r/nrArl. Inhibition of the vital sites is shown by (/) the in rire survival patterns following infection with photo- lyzed blood and (2) the in rltre rtle of inoorporarion of radioactive nuclco- sides into rhe [)NA and RNA of the parasiN. T1re developatcnt of new high affinity drugs is discussed here. Iw addnion, rhe feasibility of u,.rng slruc- tutaNy related convenlional drYp to prolect the vital site against pholoinac- tivation by FNPA is demonwraad in this paper. Speci/kally, the esperimental results suggest that the inleraction of photosensNive and conventional com- pounds with essential tarllel wes in a parasite provides a meam of determin- ing the atPinity of new derivNives of the photoprobe for those s:tes, which suqesu the structure of poremialfy useful chemdhcrapeutic agents. Torn.a4e. A. AI n d. Anndro/rAe New Yar1 Ardin.y ./Stienres)16:419-1)), 1990. From I iNron I ahoruorses, 1 rd, and rhe Univerairy of Rochester Medical Sehooi, Roehesrer, N Y SEPARATION OF D-( f )-NICOTINE FROM A RACEMIC MIXI•URE sY STEREOSPECIFIC DEORADATION OF TIIE L-(-) ISOMER rserlewwn.; pnrlla degrades .iootiee through oaidalive metabdism, but the stereospeeiAcity of this ar.echaniun lua an1 been established. In this aludy. eaperimerNa testing this sletteseketivity were directed at cwnparing absorp- tiot spectra for the metabolic eonveniom to succinoylpyridine derivatives of nMnral t: (-)-nKodss and racemic D4(t)-aicotiarc. Incubation of racanic miatures of nicotine wititi the orpwinns caused complete aereosekdive de- Rradation o( the L•(-) isoma• TM hitherto unavailable optically pure D-( 4)-okotine, which is importatM so stereochemical phumacoloRk studies of various nkotine-responsive srstems, sraa not aAecled by the bacterwm and was recoveted by ealraclion. 11 M concluded that sicotine-melaboli:ing enzymes is P. psrtife are wereoscktlive and can be used to separate the non-natural D•( t)-nicotine isoe+er from rscank rstiatures. DeTry)ia. M C and Tonren4o, A. M. Aprhid.nd fn.t.ownsewr.l Alwro6iobfl )9(5):1067-1069, 1950 I tr+m 1iu,.n I at..rNones, 1 rd. Rochester, N Y. 68 i I V1. Immunology and Adaptine Merlrani.m. NICOTINE AN f18ODIFS: COMPARISON OF I.IGAND SPI:('IFI('ITIFS OF AN718ODIES PRODUCED AOAINST TWO NIC'Ol INE (.'ON)UGAI ES Described here a an immunologic melhod for the rapid delerminatioa of nicotine in hiolosical maleriah, which ases a"semirigid bridge" for the atuchment of 6-aminonicotine lo macrornolecuks. This technique yields suR'i- ciently high levels of highly specific nicotine antibodies to study the alka- loid's fluctuating kvek in human plasma during and after cigarette smoking. Of the 25 nicotine der'wNives and meubo/ites lested, (R. S)•6•aminoaiootlne showed she highest cross reactinn. That of eotinine was less lhan 0.1% awd kss than I.0'1L /or other mnabolites. After smokinR of a single cigarette (1 2 mg nicotine in the mainsueam/ the peak blawl plasma nicotine kvek in the suh/ecls ranged from 20 to 1/H ng/nd, and the high kvels were wot necessarily found in the heavy smokers. lbis lends lo support the speeulatiow rhat individual variations in the mechanks of smoking (pun volurne, deph of inhalalion and length of lime the smoke is retained) as well as individud r.rialinns in tissue dislribwlion, metaholie rNes, and urinary clearance may have  more wMtawtial part in determining Ihe true nicotine dosage abaotbed by any sin6le individual than the actual number of cigarettes 10 or snaked per t.ni1 of time. The advent and wide availability of sensitive assays /ot uko- trne metahnlites such as described here for nierMine should facilitate a mere quantitative inierpretation of the /adon involved in the biodispositioa of nicotine. C.rno, A., Monji, N., Ali, 11., Yi, l. M.. Sowman, E. R., and MeKeraL, N., Ir. - t:uroRron /or.nd o/ aiorAendsluy I0d:))1-)10, 19tItiJ. Otber asrrprte The Ameriean Tobacco Company. From she Hormone Research Laboruory, Department of Pathok+p. Ueiw- sitr of Mumi School of Medicine. Miami. Fla.: and the Divisiow of Riochem- kal Pharmacotogy, Medical College of Virginia. Virginia Comrrwnweahh Univeraity, Richnwnd. SYNTIIFSIS OF MALF.IMIDE DERIVATWE OF CORTISOI. FOR ENZYME ('OUPLINCi IN ('ORTISOL ENZYME IMMUNOASSAY A new coni.ot derivative. corlisd-31•m•maleimidohentoae (CMS), was synthesited and caejupttd with sulfhydtyl poups of p•Ralactosidase (80). 7 his report describes both the preparation of the nr•makimidobee- zoyl derivative of cortisol and its use in Ihe development of an entyme•labekd immunoass.y (EA). lhe new mcthod was eornparCd with the conventional conjuption melhod, i.e.. cortisal•'_t•hemiwceinate (C11S/ being eonjuRMed to amino Rr.wps of A•galactosidase. 7he latter mrthod resulted in only 10'/f of the entyme being lahekd when eaamined by douhk antibody preciphation method in eacess of currisol antibody and 20% reductwn of en:yme activhy, 69

whereas the conjugation method described here showed full retention of en- zyme activity and over 95% of the enzyme being labekd with haptea derivatives 1ldh CM8 BG and C/IS-f!O conjugates had a high im- munoreactivity to cortnol antibody, and dthoukh CIIS-BG did not displace well with the added cortisol, ('MB-t>O did. Modification around tSx bridge of the cortisol derivative may have resulted in increased sensitivity to cortisol. It appean, thcrcfore, that these studie showed for the first trme the im- p.xlance of modification around the bridge for increased EIA assay sensitivity. Monjr. N. aad Cur.o, A. E,perienrl. l6( ) ) : )47-)49, F9/0. Frnm the Ilornu+ne Research Laboratory. DepartmeM of Palhnbgy. Univer- sity of Miami School of Medicine, Mrami, Fla. I)ISCF.MINAIt()N OF CONTACT At'i1VAT1ON IN PLASMA NY Pt.ASMA KAI I.IKRF.IN Thas investigation eaamined the dissemination of plasnu eoMacl activation by measuring the ckavage of Ilageman factor (I/F) molecules on two sep- aeale sets of kaolin particles One aet contained all the eompouenls of Ihe contact activation system in whole nonnd plasma, namely IIF, prekallitreir (PK/ and high mokcular weight kininoffen (IIMWK)• The second set, pre- pared from PK-deficanl plasma, held only HF and HMWK. When the particles were mieed, the HF cleavage rate on the second set wss similar to that on the first, indicating that rapid disaeminalion and the con:act reaction activity bursl occur in the fluid phase. A superoatanl factor responsible for the dissemination of the contact reaction proved 1o be taNitrein. ('kaved PK and HMWK both appeared rapidly on the kaolin aurface and in the supernate (>70-10% PK and IIMWK cleaved in supernale within 40 sec.• about 70% of each on the surface at the euliest measurement). Cleavage of PK by activated HF was at kast 17 times /sstet on the .wface than in Ihe fluid phase, since almost none oocrrted is that aa1e. Calculations ied cate a mini- tswrn PK ckavap rale of 20 nwlecuks/rer./aaolecule surface-bonnd activated HF. In eowchnioa, plnms eonlsct activation may be divided into three phases: (1) reciprocal activalion o/ s few surfnot zymoro mdecule., that is IIF and PK, with IIMWK as eofactor brinRirrR these molecules into apt.osition; (2) rapid kallikrtin release iMo the Ouid phase and eonlinued eonversion of PK 1o kaMikrcin by each twslsce-bouwd molecule of activated HF; and (3) ac- 1i.ation by fluid-phase kaUikreiw of muNiple wrface-bound IIF rnolecuks, and cleavage of multiple HMWK molecules both In fluid phase and on the surface by the soluble kaRikrein. The data wggest that the second and third steps aceount for wasl of the generated plasms eontact sctivation. Corhrent. C. G. and Revak, S. D. The lor.na/ of f..perLnrnrd Mr/kiwr IS2()):60a-619, 1950. Other support: National Institutes of IIea11h and the Office of Naval Rt- sea1, h !~..w ob Is.r.......ro ..f t...n.un..paihdoty, Scripps ( tonic and Research I.w.n./.~.n 1 . /..1i. r .1 I hF.TF(`iION OF ACTIVE KALLIKRI:IN IN INDUCFD SLISTER Ft.Ull)S OI- If1:RFDITARY ANGIOf:DEMA PAIIENTS Hereditary ankioedema (IIAE) is a disease characterized by a deficiency of Complement 1(Cl / inhibiWr funclion, usually in association with an abse.a of C1 inhibitor protein. In the study presented here, sia sudion-iwduccd bliMet fluids were obtained (rom five patients wilh HAE and eight blister (/uida were gotten from eight normal volunleen. The blister fluids were asaayed imrnuno- chemically for Hakeman fador, pretallikrein, hikh nrokcular weight (<ininoken, plasmino6en, CIa, CI inhibitor, .; maeroRlobuliw, .,-an/ilrypaie, and .,-uNi- chymolrypsin. All blister fluids analyzed coMained dNeclable arnounts of aR the proteins e.cepl Cl inhibitor. Kallikreiw was preseM in large amounls of IIAE blister fluids as as.esaed by its abilily lo IiberNe smooth-musck<oa lraclinp activity froAr purified high molecular weiRM kininoken. It was ir h.Mtcd by purrfied antibodies specifk for CIs. llte oburvalions made in this stsdy suggest that trauma iniliales activation of the /lageman-laeto.-dependcal pathways in Ihe tissues of IIAE pNieMs and produces kallrkrein aclivily. Furthermore, this kaMikreiw activity pertis/s and is unregulated in the damaged tissues. Curd,1. G., Prograis, L. l., Jr. and Coril.ewe, C. G. The Iouend of E.prrinwnrd Mtdicint 1 S2( )):742-747, 1950. Other .u'p.rtr National Institules o/ Healdr and the Office of Naval Ra aearch. From the Departments of Clinical Researeh, Immunopatholotr. and Molecular Immunolopr, Scripps Clinic and Research FotwdMiow, La lolla, Cal. DFMONSTRATION OF A PANCREAYIC PROF.IASTASE 2d,- PROTEASE INl/lflITOR COMPLEX IN NORMAI. HUMAN PLASMA Human pancreatic proelastase 2 has been shown recently to alowly forwt a strong cornples with human alpha,-prolene (nhibitor (.,-Pl) In vino. Nor, in this attempt to further characterize the molecular forms of imrmwwreaclive elastase 2, purilkd .,-P/-bound irnmunoresc/ive elaslase was partially dissociated to material having an Af, simitar~to that of proelastase 2 by incubation wilh hydrosylamine. When this fraclion was subjected to aflinity chromatography on lurtey tu white inhihitor-bound agarose, under conditions that easily separate ptoelsslase 2 lrom the active enryme, all of Ihe detectable immunoreactive elastase had the properties of the zymoten. A second peak of inununweactivt material associated with the high molecular weight fraction of plasma was shown 1o result from a specific interaction of Ihe '2'•1-Ishekd phenylmelhawe- wlfonyl elastase 2 employed as tracer in the radioimmunoassay wirh .,-macro- gIoMdin, resulting in apparent immummactivNy. llrcse results denionsiratc that 111 1 71

0 all of the detectable immunoreactive pancreatic elaslase 2 in normal human plasma is proclastsse 2 biund to .,-protene inhihitor. I ar6man, C.. Sn>,lrick, 1. W.. Geolar, M. C.. Johnson. 1. N., and Fasselt, M. Anvrk.n Iournd of fhrriolopy 2)t()):G177-1U2, 1960. OtAer errprt: Medical Research Service of Ihe Veterans Administration. From the Fnzymoiop Research Labora/ory, Martinez Veterans Administration Medical ('eMer, Martinez. (-al.; sod Ihe Department of Internal Medicine. Umversny of California School of Madieisn, Davis. At 1.ERGY TO TOSACCO: AN OCCUPATIONAL HAZARD Sentitivily to antigens in tobacco may ewse allergic reactions in humans. Presented here is a case history describing a paliem who developed rhindis and breathrng drfl.culry only when she worked al a citarette factory. Nurncrous test results indicated that she was alkrgic to green lobaceo kaf antigens. //er re- action to cured tobacco leaf was only borderlitse, however, probably because of a lower antigen concentration doubkls resulling from protein denaluration during the curing process Ahhough tobacco smote also contains anligem, this patient farkd to resc/ eonsntently to both inhaled cigarette smoke and tobacco smolie lar estract Furthermore, her IgP- antibodies did nol react tn tobacco glycoprotein when serum was lested directly with a solid-phase glycopro/ein or when thn was uscd to inhibit their binding 1o sdrd-phase green tobacco in 1he radioalkrgosorhent test lherefore, this patienl was not alkrtic to tobacco smoke. these results confirm urne previous reports that allergy to tobacco producls is aw occupational hazard /or worken i. this industry. C.kkM.G.I.rr.l TAe New Enelend /o..nal of Medicine )02(11) :617-619, 1960. OtAer aurr.rt: National Inailutes of Heahh and the Mayo Foundation. From the Deputmenls of Medieine. Inunwwlokr. and Pediatrics. Mayo Medical School. Mayo Clisic and Foundation, Rochester. Mire.• and the McGuire Clinic, Richmond. Va. APOI.IPOPROTEIN IS RFSPONSISLE FOR NEUTRALI2ATION OF XENOTROPIC TYPE C VIRUS SY MOUSE SERUM tnurine lipoproleiws coMain a po4eM inhibilor uf /he mnuse lin l Ci ~ rcu a senotropic type C.irus and this aeulralizing activity (NA) does no1 involve immunoglobulins or complemeM. According to fractionalion sludics- 1he NA is primarily found in the Iri6lyceride-rkh lipoproleirn (prcdomisaMly chylo- mkrons) and in the /1DL, subtraction of the high density lipopro:eins (1/DL). T1ese results indicate NA is astweiatcd wilh panicles of the same flotation properties, disreelen and ekctrophorelie charges as the serum lipoproltins, and with more than one clsss of serum lipoproteins. When mice are (asled. NA n found mainly in the 1101- (rsclion. During alimentary /ipemia, both the Iq.opnwcins of the < I 0(Mg/cm' fraction and the HD1. show a striking clevauon in nrairahrauon tilen, which persists in the IaNer aller the lipemia 72 I hac receded. Ihis transfer asl NA hetween ND1. and very low density lipo- prolcin. in the mnuse can alw be demonurated in vitro. Removal of lipids frum the serum lipoprnleins by Ireatmenl with letramcthylurea significantly Mwers NA but the apolipoproleim remaining nlain sonre NA, which is ea- hanced by their binding to a phospholipid-slahilized 1rillyceride emulsion. NA is aholiched by proteolytic digestion and aMiserum apinl HD1L eliminates the NA in all lipoprotein fraclions. Theae resuhs indicate that senotropic .irw neutralization by normal mouse serum relies on a protein which can migrate amonk variorn lipoprolein particles in a manner analogous to thil of other mammaliarkapolipoproleins. Because these v'auses are t.biquilous and associated with both normal and malignant lissues, the inleraclion of endogenous type C viruses with circulating Iipoproleira may represeM an important hou regulatory mechanism. Kane, /. P., llardman, D. A., Dimpfl. I. C., awd [-evy, /. A. ProrrcdinXs o/ the NMlowal Acdemy o/ Selencn o/ the Unlrrd Srwn o/ Amerk.r 76(11) •39S7-S%1, 1979. Otlrer saerrortr National Cancer Institute and the U. S. Public Ileatth Servia. Front the Cardiovascular Research lnslilute, Department of Medicine, and Cancer Research lnstilwe, Univenily of California, San Francisco. ENDO7IIELIAI. CELIS OF BOVINE PULMONARY ARTERY LACK RECEPTORS FOR C'3b AND FOR THE Fc PORl1ON OF IMMUNOGI.OBULIN G This study attempts 1o clarily any etislinR role of pulmonary endatbeR.) cells in the binding of eirculating itsnnuwe eompleaes. Specifically sou6bl was the preseace of receptors for the Fe prtiow (ery/slrllizabk frakme./) of IkG ( immu"lobtalin G/ and for C3b ( tb component of compkme.t ). Rewhs show that bovine pulmonary etwlolhelial ceRs in cWture do not possess 111ese recepton, a fact whicb is nol surprising in view of these ceNs' prinary, futrclion, which is to provide a unooth, filiding aurfux for hlood flow. The lack of these receptors rnay. Iherelore. help eaplain 1he etdahelial ceRs' aonthrombogenic funclion. It is evident then thN end0ahelial cells do aa1 pardcipate in Ib mechanism by which the soluble inweune eotwpkses can pin access to the interstitial space of the hnlp either by way of receptors for C)b or tbe Fe portion of 1s(3. These reauNs, bowe.er, do wu1 rule out the possibility thM other compkmenl cowtponenls may danade or olherwise imeraet with ew- dothelial eeRs to irwluce iwtravnculsr thrornbosis. It appears thM obtaining Ihe rekvant monospecilk aMibodies is the rale-limiing step iw the elarificatio. 0 the role o/ enduwhelwm in henwstasis. the processinR of hormones awd the Iransprxs of mclaholiles. llran. U. S. to .f. Scirnre 20t1:71N-749, 1980. OOther s.rrp.ra: U. S. Public Ilealth Service and I/artford Foundation. 1'rom the Ikp.rlment of Medicine. University of Miami Schorrl of Medicine. Miami. Fla. 73

THE REGI/l_ATION OF T1IYM1DINE SECRETION BY MACROPHAGFS In the in vitro studies reported here, macrophate rnonolayen were cul- tured with minimal essential mednrm (MEM) or L-cctlconditioncd rpedrum for up so four days. lhyrnidine kinsat (TK) activity and sil thymiJine in- corporation by the celk, as well r aecretio, of thymidine into culture auper- natanes were assayed daily. Resuhs showed 1ha1 macrophales cultured in MEM released thymidine and did not •aprer TK. However, when macrsph•6cs were Incubated with medium coeditb•ad by L oelh, they eap.essed TK, (n- coporaed sH thymidine In/o trkAlore•atk acid precipilabk materul, and ceased to .ecrete the wuckoside. Pwtherware. repticatM6 PI)1f/D1 cella were Indrucd to secrete thymidiwe by i.Nbiliry TK with d-ghrcoeamine. These studiea have ahown a conelniow between thrrnidine aecretiow and the absence of 1K activity bosh in normal macroph.6es and P)@6/1)1 eelb inhibixd with d gtucosamine. Converselr. no thlmidine wa deiected in supernatams Irom rrucrophage rno.olarers treated with growth factor and (rom P)Iit/l)1 celh, bdh of which espreaed TK activit7. It seema, therdore, that the secretion of thymidine by macrophages may be eaplained by the lack of TK. Stadeeter, M. 1. and Un.nw, E. R. The lorrnd o/ /mmrwology 12)(2):361-371, 1979. O/Aer a.pp.rtr National Cancer Institute. From the Department of Pathology. Tufts Universily School of Medicine; and the Department of Pathology. Harvard Medical School. Boston. lT1E SPECIFIC BINDING OF LISTERIA AfONOCYTOGENES-IMMUNE T LYMPIIOCYTF.S TO MACROP/IAGES. 1. QUANTITATION AND ROLE OF 11-2 GENE PRODUCTS fased on the hypothesis that T cells recogniz,e anti6e. associated with Ir 6ene producM, these studies carefully esamine the role of TteR-macropha6e contact in T-cell activatios and Mlempt to detessni.e if Ir lens products dic- t•te the anti6en-specifk physical MNeraction bewee. T cells •nd macropha6es. The apstem developed here allows testing of the correlation between direct rneasurenxaa of activation br qrtantitating specific T<eM-maeropha6e bind- ing. Specific binding, which was apparent •fter • brk/ (I hr.) contact, waa measurcd b7 the depktion of LLseria monoc7roRenes-speeiBc T-cell activity in the cella nnnadhereeN so L. wsonoryrocirnes-Pulsed macrophate monolayers. The L. nwweryropnrs-speci/k T-cell function was determined by its ability to aetivate L, nwwoc~ro~enes-pvlsed macrophages to seerete a prdein mito- genk /or thymocytes and to effect nonspecific tumoricidal activity. These T-cell function manifcslMions are re6ulated by products of the 11-2 gene compka I region According to the resuhs. H-2 gene products have a direct rather than just a triggering role in mediating the specific binding of T cells lo macrophages 'I his also implies that the antigen dependent physicd inter- actinn between I cells and macrophages is the initial and delermining event in some types of 11 2 6ene controlled immune reactivity. 7iegler. K and (Inonur, F R. I I The lorrnd o/ Ezperimenrd Medicine 130:111I-1160, 1979. Other aupportr National Inslitutes of Health. From the Departmenl of Pathology, Harvard Medical School, Boston. A DEFECT IN TIIE ANTIOF.N-PRESENTINO FUNCTION OP MACROPHAGES FROM NEONATAL MICE In the tiswe cuNtrre o.uy ayst•rw teporud hns. T Irmphocyt•e from mke Mnnwnized •pirt L/srerie n.owarpre6twrs were evhured with aaacre~ pha6es which had taken up heat-liBed Lisrrd" oqas*rwa. Whes the Liuerio- imnwne 1' Irrnphocynes Irorn aduh (2-I-mowth-old) mice were cultured witb 1/sre.i.•pulsed macropha6es also from aduhs. T-cell lymphocyte proliferation resulted. Ilowcver, only a weak prolilerative respowse was oeen when the same number of macrophyes from rsconatal mice was aubMilwed for the macns phases from aduhs. In faet, the ability of macrophyes to inkracl effectisely with T-lymphocytes did nm occur erntil 3-4 weeks ./kr birth, approaimMelp the tirne of weaning. Aho investigated were the effects of T-lymphoc7te-macre- pha6e imeraclion other than proliferation. Restdb showed that although wucro- phages from adults secreted a special miloges tor thymocyles when wdsad with Lisrni.-immuwe T IynqhocYtet and Lisrerl., macrophages from neo.atal •nimah did not. Ako, the macropha6e pprlNioa /rotw neo.Nal wdce was ahoww to be deficient in a wbpoprlNioa carrying surface Ia antiRew. O.eraN, then, this study demonstrates thN macrophages from neonatal mice do not present antigen efficiently and that this can be correlated with the small aww- ber of macroplu6es bearitrs the I• antigens. Lu, C. Y., Calamai, E. G. and Un.wne, E. R. Narrre 292(5716):327-329, 1979. Otker aopp.rts National laslitutes of Heahh. From the Depanment of Patholop, Harvard Medical School, Boston. a EVIDENCE TIIAT TIIYMOCYTES REQUIRE AT LEAST TWO DISI INCT SIGNALS 10 PROLIFERATE Accumulated evidence indicales that at kaM two atirmdi are resluird to initiale and mairwain proliferation is srowth-arrested Rbroblasts, and thM. M 6eneral, optimal prolderatinn maT involve wwltipk induceive si6n•b. Thi report is based on attempts to eaamine the node of action of a thywacpte- powth-pronwting nsokcuk elaborated by macrophqes. Macrophape culture fluid ( M('t^ ) contains a 15,1100 dahon prokin that preferentlally stimulates thymncyte proliferation. lhese studies indicate that the optirnal receptlvity N thymocytes so the mito6enic stin>tAus in M('F is limileJ to the time drri•d culture when these cells are prulilerating spontaneously. Restins eelh c.n be "activated," and their M('F respnnse taally restored, by a brief wonmilolicnic kctin pulse. This system provides a model for the evaluation of tAynaqrt• proli/eration, particularly the meehanisrn of action of MCF. It ia propo.ed 74 1 75

i 1ha1 M('1' supplies a quantdativcly distinct signal that acls only on aclivalcd lar6et cells, a sure apparcntly inherent to spontaneously cycling thymocytes. 6eller, 1) 1. and Unonur, L. R. ] Ac lournd u/ Invrnunobry 12 )(6) :2190-21t9), 1979. Other arpportt National Instilutes of Health. From the Department of Pathology, Harvard Medical Schod, Boslon. A('('IiSSORY Ctil 1. RF.Ql11RF.MENT IN T/IE PROLIFERATIVE RESPONSE OF T LYMPII(X'YTFS TO HEMOCYANIN lhis rtporl dc.cribes a sludy of the prolikraion of T IympM+cyta, taken from mice irwmunc to the large polymeric protei., keyhole IimpN hemo- cyanrn (KL11). Once obtained from the taioe, the T cells wete ksled in cut- ture for Iheu prdrlerative response to KLH or so eoneanavalin A. The reac- tion lo K111 was dependent on the presence of la-bearing usacropha`es, as shown only after esumrve manipulaliow of the T<eNsdepktinR accessory cells by adherence to dnhes and passage through nylon-wool columns. Macro- phalle dependcncy requirrd homology with rnacrophages Irore mouse strains sharing rhe left side of It 2 and was o01 bypaued by conditioned media from macrophagcs. On the other hand, the proliler.tive response to eoncanavahn, which was also mactophage dependeM, did not display 11-2 reslrKlrM and eoutJ be rrplaceJ hy a macrnphage<nnJN.oned medium. Nonadherenl spleen etlls, eslensively lackrng in macrophagts, could substilule (or macrophates but at a 1011-/oW escess. ('ells isolated with anti-1g reagents in the fluorescence- activated cell sorter also had an antigen. preseMing functi.m. Whether these results imply an anagen presentmg funclioo of Seelh is the queuion. Kaesmer, G. MW aed Un.nrr, E. R. Clinkl/mmrnolosy.nd lmrnrwor.aAobPy 13:1)1-4I), 1950. Olhrr aopp.rtt Nalional Inslitules ol Health. From the Ikpartmenl of Pathology, llar.ard Medical School, Boston. REGULATION OF MA('ROPl1AG1: POPULATIONS I. PREhFRENTIAI. INDUCTION OF Ia-RI('H PI:RITONEAL EXUDATFS SY IMMUNOLOGIC SI IMUL1 In this initid report of a scries of studies iavesliplinR factors Ihat may regulate the la phenotype of macrophage populuioes. the amounu of la-posi- live and -negaUve macrophages were noted in peritoneal eaudates of normal mice or of mice (1) injected with various inRammalory maleriab, (2) infected with f.lsrnie n.oworylotrnrr. or O) Injected with hemoeyania. Results showed that L-neplive macroph.ties predominated in esudales from nurm.l mice or from those given .nrnrral oil, peptone, thioglycolate, culture media, or endo- L-srn On Ihe .dhrr h.nd, infection with fJrlrria caused a very marked in- crea.e in la prnui.c macruptuRes. Ihe induction of la posilive maarophages 76 hy l.i.rra.. inoculation resulted in grcal part from an immune process. The Ia-pusitive etudales were more readily generated in immsrne mice givca a secondary challenge with heat-kilkd organisms. Furthermore, immune T cells transplanted together with heal-kiNed orpniyns into normal mice resulted in la-rich esuJates. Injection of hemocyanin also induced la-rich eaudatn Involving an immune process. llrese resuMs isdicale that there is a diAerence in the relative content of la-positive macrophaRes In peyitorte.) eaudNe, dter diRereni kinds of alimuli; and dso. that those tlwrwli generating eaudates highly enriched for /a-posilive cells resuh, at kast In part, from an lmmuno- btic process. Also, the observation of T cells inducieg etudales enriched in la-positive macrophates represents another segment of the complex T aN- macrophate relationship. In essence, Iherefore, these brlinss show that d- Ihough both immunodenie or wonirnmurr.Knic pjmuli lead to an increass in periloned esuJare ce1M, h is only when arNillew-speciAe T cells are involved Iha1 there is a signilkaM and selective enrichment in la-posilive macrophates. delkr, 1). 1., Kiely, 1-M. and Uw.wre, E. R. Thrlorrndo/Lnmrnolopy 121()):1126-11)2, 19190. Otber wpportr National Insleules of Health. From the Department of Pathology. Harvard Medical School, ROs1on. I la ANTIGENS AND ANl IGEN-PRESENT/NG FUNCTION OF THYMIC MA('ROPHAGES After enrkhnsenl on dencily padinys, ns.crophaRes were the predomi- w.nl ceR type found amons the iwlaled Ihysaic adhereM cetls, as established by morphoMrdy. Fc uW f I surlace receplors, phadoeylo.a, and eslerase ao- tivity. In addition, a minor fraction of cells with a distinctive dendrilic mor- phology had surface properties similar 1o /hore of esacrophages, but lirwited pMtocytie capacity. About SOx of thymic adhetesM .eNs carried la antigens, as shown by immurr+lhrorescence and Ihe rse of either A.T11 anti-A.TL or nanocbn.l anti-l-A anlibodies. These cells abo proved to I,e an eslremely e0ea live antiRen-presentinR aouree for euerophaffadelsleled iwwtwne T eells, suppors. ind the noliun that Ihe la antiffens delecled are funclionatly siRni/lcaas. Theu data suggest that the nsacrophage, isolaled by physical disruption of the /hyrswt or hy c.dlagena.e JissociatirMS ol its slroma, is the predominant adherent eeN type and the principal la-bearing ceN. The thymic mxrophages am1 the asso- eialed pr>p,d.nwn of JenJtilic crNs which ean stimulate lhynrucytt mar.nration and proli/eratirar in rn..., may reRrd,at eerlain uales o1 1 cell developmeM in vir•o. It i. Irrpcd that the ccllul.( iMcradiAns Rovrrning T cell diAerenlia- lirws can be brtlcr d,Kned by .eparaling the two ecll types so 1ha1 their lune- Iiun nray he evalualcd irwkperwl.ndy. Iklkr, 1). 1. and t/nanrr, h'. R. The lournul u/ Iinnrrnnlury 1244 1) :1 4))-1 a10, 19110. O/Arr arpp..rt: National Inalilu/es ol Htahh. Frons Ihc /kpartmcrnt ol 1`.rhokrRy, Harvard Medical School, 8oston, 77

T CEI.1.-MACROPHAGE INTERACTION IN INFECTION TO 711E INfRACE1.1.U1.AR PATII(XiEN LIS)ERIA MUNUCYTIN;ENt:t Resislance to t.urerid monoryrogewe:. a viruknt pathogen for the mouse, depends on activation of Aoth 1 cells and macrophates. While Ihis resistance can be conlerred upon nonimmune mice by transplanting synkeneic T cells Irom immune mice. it appean now IhN the transfer of resistance to t isuerid infection takes place best when the dowor ot the T cells and the re:ipient share the I-A region of the major hiaocompNibility compka (MIIC) of the species• that is. The murine 11-2. la the iw .Nro caperireents sununariial here, I~rre.i.-immune T cells aed macroph.pes fratt woninwnune mice were mised in culture topether with heat-kilkd Ltraria orpniuns. and the followi.= three parameters were evaluated: (1) secretiow of bioloAically active mokcuks by macrophaAes; (2) T cell proli/eratio.; a.d (7) activuion of rnacropha6es for eytocidal activity Resrlts of these etrdio showed that the lollowing things are necessary lor inductan of the three phenomena: (1) speciAc tiuerL- immune T celh; (2) addean ot antigen, in the form of dead Lirrerm: (7) homology at the I-A region between the T cell a.d The macrophage; (4) macro- phages bearinA /a anti6ens; and (5) eell-tocelt eoouct. ~ Un.nre, E. R. In: l/nanue, E. R and Rosenthal. A. S- (eds.): Maropilajor Resularion o/ lnrmunrry, New York: Academic Press. Inc.. 1980. pp. 7)-eS. Other aupportr National Institutes of Health. From the Deparlment ol Pathology. Harvard Medical School. Boston. QUANTITATION OF (:I:NI:TICALI.Y RFSfRICfEI) T CEI.I.- MA(-RUPIIAGE BINDING Numerous qudies have established the role o/ 19 genes in the control of immune resporaes and, rnore recently, have provided strong evidence of a central role /or the macrophage in the eapresaiow of Ir gene /unction. Based on the hypothe+is that T cells recoRwise antipe. in the eodeal or It kene products, the work reported hete atterwpled /o evaluate the role of T cell- macrophage eontact in T eeM activatim asd to dekrmioe whether It gene prod- ucts diclate 1he aMi`cn-speci/k physical Interaction between T cells and macro- phakes. To thii end, a system was developed so quantitate the antrken-speciflc binding of T cells to macrophaAea as an analog of the initial phase of I cell macrophaRe activation: antigen reootoiuo.. This appears to be a suitable model /or thr purpose. The results obaiwed here, together with others /rom guinea pig studies, strongly suggest that specifie T eell-macrophate binding is the initial and determininp eveat in H-2 Rer control of imnwse re.nivity. However, neither the mechanism by which anligen recognition by T eeRs is accornplished, nor the nse.ns through which It gene /unctios is eapressed by the macrophage. is deAned The hope is that the binding system described here will permil a systematic and definitive analysrs of the 11-2 gene-direeled clonality of the 7 tell r.rcrr.wre. at -eti as pro.e usc/ul to study the biochemical events in- vnlvcd in she handLnt ' ul antigen by the macrophake. lirglcr- K and Unonuir. 1. R I In: Unanue, F. R. and Rosenthal, A. S. (eds.): Macroplhaze Retul.rion o/ Lnwrrnrrr, New York: Academic Press, 19t0, pp. 245-262. Otlier .upport r National Institutes of Health. From the Department of pathology. Harvard Medical School, loston. R()LF OF MACROPHAGES IN THE REGULATION OF THYMOCYTE PRULIFERA I ION AND DIFFF.RENTIA7ION An evaluatiun of IymlrhostinwlNOry /nctors from peripheral rn.crophaRea and, more receetly, of the ability of thymic rwacrophaRes to Ihewaelves reRrt- late thymocyte development in virro, has led to a broad eoacepl of rnaerophaM lunction. While it has bcen widely accepted /ha1 anacroplrapea are a critical regulator of T cell activity. the studies reporled here suggest that they tnay dso play an equally imporunt role modul.tinR the prdi/eratio, and ddere.- tiation of thymocytes. While evaluating the ttmure of the /hymocyte response to the mitogenic activity in macrophaite wMure Illnid (MCF), i1 wY found that thyewcytes responded relatively poorly to McF i/ initial eaposure was delayed 2/ hours or more. Optimal response was restored, however, when MCF was preceded by a brief, essentially nonmilogewic pulse of Con A, iadi& catinR that the re+ponse to MCF is based on a drd signal mechanism. Also, N was found that peritoneal macrophaRes can secrete a(AymocytediAereo- tiatinR factor which prornaes at kasl one seflirenoe of the T ceN developnesN pathway. Theae Andiap led e.perimeaally to the isolalion of macrophyes /rom the thynws and to the evahution of their activity. Rewha showed thM the macrophaRes slimulaled developmental ehanRes similar /0 Ihofe induced by the diHerentiatinp (actor from peripheral wncrophaRea. In addilio., Ihs AndinR that they are strikingly la positive fwther MrenRthens their poleai.l role in the reRulation of T cell development. OveraM, these observatioam oor- pkd with preliminary evidence that thymic maerophages. in the absence o/ esokenous sliorrli. actively secrete a mitoRenie /aclor. suuesu that the thymie macrophate may be especially adapted roo a role aa am efreaar of thyn,oeyt" proliferation and dillereMiation. tieller. D. 1. and Uw.nre, E. R. In: Unanue, E. R. and Rosenthal. A. S. (eds ): A4acropA.w Regu/.rion ./ Irnmrrnir New Y k• A or r d . a e . c mrc Preas. Inc., 9920. pp. )61-)7t. Other support: National In>tilutes of Ileahh. From the tkpartmcnl of Pathology. Harvard Medical Schod, soaon. THE DIFFUSE NEUROENDOCRINE SYSTEM: S7 UDIES OF THIS SCOVERED CONl R O1.1.1NG SYSTEM IN IIEALTH NEWIAND DI.Y DISEASI? /n 1931- i1 was ahown lor the Arst lime that an identical peplide was present in both the brain and the Cut. Now, nearly Ave decades later, an ea- pbsively irkreasing number ol peptides are being lound to be common b

these two siles, including /he following neuropep/idcs: somstostalin, enke- phalin, gastrin/C('K, hrNnbesin, and vasoactivt intestinal polypeptKfe. lhese pep/ides awnpose what is now krawe as the dlAuse neuroendocrine system. 7 wo immunological methods, radioirnmunoassay and imnwnocytochemistry. were useJ in combination here to study the distribution and cellular localiza- tion of these neuropeptides. A number of these suhsuncei have been found to he ahnormal in the disease slate. thus providing further informatinn as to their rilfe in normal and pathological condilion+. Ihe Andinp reported in this paper indicate clearly that the control of bodily functions is modulated by numerous lactors acting in  synergistic or antagonistic manner. It is thus totally irnufficient to analyze, separNely, one singk factor; a multidisciplinary, muhiprofile approach is what is needed. Sipli/kantly, the esplosive and ea- ponential discovery of numerous neutopeptides. all with powerful controlling actions, and ow increasing knowkdge of Iheir distriburion, are beginning to open up avenues (or new ways of underuanding the compkaity of coMrol of body lunclions and the unbalancing of this control is diaease. Polak, 1 M and Bloom. S. R. (WYf, !. A.) Thr lorrnal of Nirrahrr^irtry and C)'tocAewd+ny 27(10) :1)9d-1 400, 1979. Other ..pport: Medical Research Council. Wellcome Trust, arn1 lenssen Pharmaceutical. Ltd , Londo.. 1'rom the Departments of Nistochrmistry and Medicine. Royal Postgraduate Medical School. /lammetsmqh Ilospdal, I.ondo.. V11. Epidensio/ogy TIIE REI.ATIONSIIIP BETWEEN MATERNAL SMOKING AND T11E INCIDENCE OF CUN(;ENITAL ANOMALIES Esamination of 14.735 children, members of the Kaiser Foundation Ileahh Plan. for the incidence of congenital anomalies in relation to maternal smokinR reveakd no signiAcanl dillerenees when smokers (all dose levels combined) were compared with those who had never smoked. Mothers who were paM smoken were also included in the study. DiAerences did emerge. however, when the anwrnt of smoking was taken into consideration: the inci- dence of congenital anomalies decreased slightly among the oAspring of light smokers; but a significant difference in incidence appeared when the offspring of mothers who smoked 20 or more cigarettes a day were compared to the children of those who had never smoked. The increased incidrnce associated with heavy sn.o\ong was predominantly seen among male cdisprin6 and re- flrcrrJ r-•Mr•/r •t~rr^al.nws Iwu sprcdic onrs, however, rnguinal hernia •MI .r.,^, sr.v.• c.een.A..ndy in the gruup of chddren whow murhets h..l .....4.J J.....•t r/1n IM dhrr hsnd the rncakn.e ol mc>,terate /1r1..ns. i 1 mu.cah/skcktal nomalies decreased aijnificanlly among the male offspring of .nurkers, parlicularly litht srnokers. These dals, as weR as the overall un- ususl Jose-responsc rtlalionship, convey the impression that the associatiom observed here are caused by different and comple. laclon. Further invealiRa lion of these is suMesled. Christianson, R. E. (ran den errP, R. J,) American lournat of EpJdrmiolory 112 ( S):681-695, 1990. Other suppertr National Institute of Chi1d Nealth and Huaaa. Development. From the Child Health and Devebpment Studies, University of California School of Public Health, Berkeley. ('I(iARl:/'1 E SMOKINO, A(TINO, AND DECLINE IN PULMONARY FUNt:.'7ION: A I.ON(IITUI)INAL 51 UDY A previous crors.sectional study, which eaamined the relative eAeeta of aite and smoking on pulmonary funclion, showed that both variables eem- Iribute to lower forced vital capacity (FVC) and forced eapiralory volume is one second (FEVr.). The same spiromelric da/a showed that in thoaa who inhakd large vdruees of am+ke, these pulmonary function parameters were significantly lower than in those who inhaled small vohnnes or who had never smoked. This study attempts Io eatend /hese observations by using a bql- tudinal design and also esamines Ihe relationship between pulmonary frnclion and a quantitative iwdea of smoking hased on the actual lar sed wicotine in- lake of smokers. Consequently, age, lar anJ nicotine consumption were evab- ated with relation to decline in pulmonary function /or ewreM, toemer, anr nommokers, as described in the present report. Three serial apirometric FVC ard FEV. „ determinations were dnne at specific istervah during a period of about /o years. The subjects included 268 adult male cigarette smokers, 1I1 quilters and 254 who had never smoked. The Arst group was divided hne high- and hnv.tar eonsumplion caleRories. Data w(rc analyzed by repealed measure analyses of variance using time (aging) and smokinR slalw as inde- pendent variahles, and age as a eovsrisle. Results indicate that FVC .nd FEV,, are related 00 /muking slsqn. In aR age poups, smokers per/orsned the worsl on spinrmetry, while nonsmokers performed best. Curren/ amokers. especially thc high-ur Sroup, showed the greatest decline in Fi?V over the eonrse of 1ime, but bss of FVC in telaliun 1o time was nut inKalced by smokins status. Aotsi. R. rt at. . Arcli(rrr of Enrironrnrnra/ Nrdrh 7S(1):?JV2S2, 1950. Other aupport t Medical Research Service of the Veterans Administralio.. From the Normative Aging Study, Veterans Adminis/ralion Outpatient Cliwic, 1lnskMl; Ilelknic t'solkec. Rrnokline, Mass.; Univcrsity of Mas.achuselts, f1loa- lon; Pldmonary Unn and (:cr.rmology 1)ivision, Ihorrwhke I ahoralo.y, Beth Hracl IluspAal, and 11.rvarJ McJkal Schtxll, Roslon. NII I MI

Tllli INFI.UEN('1° OF A(:1°., AI ('O1101-CONSUMPl lON, ANI) BO1)Y BU11 D ON GONADAI. fUNCt ION IN MFN In this threefold stndy, basal plasma levels of testosterone, dihydrotcs- tostrrone, estradiol and gonadotropins. and testoslerone binding capacity (pert cenl radioactive tcstosterone hound to protein) were measured in healthy carefully screened young ()1-.. yr old: n= 44) and older (M tlN yr old; n= e?) male participants in the Normative Aging Study of the Velerans Administralion. Results here showed 1ba1 plasma testusterone levels were not influenced by aging in normal tssew. Then was no slatislically si6nificanl effect of age on either teslowerone or tha free Iesloalerone imlea, although the tcs- losterone-hinding capacity was higher in the older :roup. Of the two products of testosterone metabnlnm studied, estradiol did not change with age, while dihydrulesloslerone was lower in the older group. The follick aimulating ho.- mone kvels were increased among 11se older asen. hut luteiniring hornwsne levels were not significantly inllurneed by age. Aho. Ihe level of chronic stable alcohol intake had no effect on gortadal (uwNion, as estimated by teslosterone kvcls and the free teslosleasne indet. Finally, analysis of Ihe relationship be- tween b.dy buiW and hormone levels indieated that estradiol levels were highest in gynandromorplsic men and lowest in nsesonwrphie men. Sparrow. D, Qon1. R and Rowe. 1. W. Iourn.d o f Chnk.l F wJor.ir.olopy and Metabolism S 1( )): S0R-312, 19110. Other aupporf: Medical Research Service of the Veterans Adminislralion. From the Normative Aging Study, Veterans Administration Outpatient Clinie, llelknic ('alkgc, and the Geronlology, 1)ivision, llwrndike Laboratory, Bdh Israel Nospital and I/arvard Medical Schod, Boston. SMOK1Na MOTIVE FACTORS: A REVIEW AND REPI.ICATION Some consensus on the number, tulure and function of smoking motives or styles seems to be emerging. As yet, however, large-scale studies on the stability of nnNives and the utitity of the smoking motive model have not been reported. This paper et.rnines the psychometric questions of faclorial replicabwliq and longitudinal stabilily of the motives. In 1977 anf in 1976, qtrestinnnaires (/lorn-Wain6row Snmker Survey plus 20 additional items de- ve/opcd by Coan) were sent to 1.340 current and former smoters (aduN maks). Factor analyses determined that the original factor structure was sat- isfaclorily replicated hoth times and that addition of the nesw items contributed only one rtew factor. The 3-year stability coefficients for hoth current ,ad former smokers ranged from 0.45 to 067. 'TAese results are consistent wNh the notion that smoting motives are a relatively stable refkclion of person- ality train lhe reliahdity of the mcasuremenl method eslablishe.l here also prnvidrs the basis for future research concerning the validity anJ utility of this apl+rnach AJnunistralwsn of the type of questionnaire used lure is rapid, simple and inrapcn.eve and all investi6ators evaluating programs aimed at sirnokmg .essatam .hootd he encouraged to adopl such measures and to analyze I the daa within smoker types. 'this could lead to the ulili:alisn of smoker mo- Itve mcaWSements as diagnostic aids in choosing the roml appropriate regimen for an IMhvldual. Couo, P. 7'., lr., McCrae, R. R. and SussE, R. Tlir Inrrrnnrionaf Journal of rAr AddinJons 1S(1) •S)7-319, 1950. Other aupporfr Medical Research Service of the Velerans Adminiuralio.. From the llniversity of Massachusetts. Boston, and the Normative Agie6 Stwly, Veterans Administration Outpatient Clinie, Boston. INt I UEN('t:01: f?XTRAVtRS1ON AND NEURO7ICISM ON SUBIE('l IVE Wt:1.L-et:INU: IIAPPY AND UNIIAPPY PEOPLE Personality descriptions of happy people generally reflect psychological and social adjustmeat. while descriptions of unhappy people may often mirror signs of neuroticism. Three sludies are reported here that etamine the rela. /ions between personality and happiness (or subjeetive well-being). In Sludy 1, the relation hctween four measures of happiness and seven personality dis- positions hypothesized to be related to positive or negative affect is etamited. In Shdy 3 an atlempt is made to elarily and organize these results using measures of the broader dimensions of ealraversiow /E/ and neurolicism /N). and in Study I happiness is predicted from E and N data obtained 10 years previously. Results of these studies support the hypothesis that one aN of disposiliuns is responsible (w positive affect or aalisfaclion, whereas anolher, wdependcnt set of dispositinm infiuewces negative aBeet or dissatisfactio.. The data from Ihese studies eAecti.elp rWe aM on ahernNive etpla.atioo /hN associations between happiness and personalitr result solely from the tnedi.t- ing effect of temporary moods or slales. This AwJieg is also impressive r ind'aeN evidence of the enduring efleds of /hese dimensions of persondiy. A model of individaal differences In happiriesa is presented Aete, .wid tAe separate and compkmentary roks of Irait and adappation.kvel theories in et- plaining happiness are discussed. Coua. P. T., Ir. and McCrae, R. R. !o«rnul of Prrsonaflry .wd Sociuf Piyc6ofokr 7R(d) :661-67/, 1950. OOther support r Medical Research Service of the Veterans Adminislraliow. Ft -im the National Institute un Alging, National Instilules of Health, fadUmore. a A SYNTIII:TIC BASIS 1()R A COMPREIIENSIVE FACTOR-ANALYSIS 7111:OR Y The postwar development of three faclor-analysis theories, which IoplAer form a basis for further refinement of Ihis subjecl's melhadology, is reviewed R2 I R1

here in the hope of familiaririn6 an increasing number of binmelriciam with ih specialized developments and thereby broadening its potential. The all- important points JiscusseJ here are summarized as: (1) the lormulation of the /actor-onalysis model as a special type of secund-order slochastrc process. (2) the masimum-Irkeldrood theory of cstimation of variabks in the factor- analysis model ie Ihe face of a normal disuibulan of response variates, aod ()) the theory of analytic rolation based on optimization of an objective function. The subject of rotaliau fs a unique feature of factor analysis and essential to it. WilUams, 1. S(C.rmperlrr, n. W.) eiomrr.ks )S(4):719-7)), 1979. From the Department of Statistia, Cobrado State Universitr, Fort ('ollins. STABII.tTY OF A FACTOR-ANAI-Y71C DESCRIPTION OF SMOKING BE:IIAVIOR In this widespread ttempl to betler undentand the eompks human be- havioral trait of tobacco use, questionnaire forms were adminntered to urban- ites and suburbanbes of mised .pea in a part of metropolitan (knver; college nudents in Fort Collins. Coiorado; middle-aged urbanites and suburbanites in Ihe rnNropolitan areas of (a) Stoekholm, and (b) Goteborg, Sweden; and the adult children of the Swedish groups (a) and /h)- Factor analysis of the questionnaire responses idenufied three types of human smoking behavior which eshrbit a stable structure Type I smoking behavior was closely related to a personally perceived need foe tobacco producls. Types 11 and I11 were related to prychosocial-sensorimolor rewards and Ihe intensi8ca1-an of pkasure, respectively. The typology was easily detectable and remarkably constant is all the populations studied. This stability of the factor slructure would sull- 6est that the factors are real in the sense 1ha there eaist widely shared motives for different types of smokint behaviors• WiBiatsa,1. S., C.rmpac4n, D. W...nd Krier, M. 1. Drue .nd AfroAol Dependence 3(6):467-I71, 1980. From the Department of Slatistics, Cokxado State University. Forl Collins; and Department of F.nvironmenlal, Population and Orpnismic Biology and Idstitue (oe Behavioral Genetics. University of Colorado. Buukkr. A TWIN MIiTFIO17nLOOY FOR THE STUDY OF GENETIC ANI) ENVIRONMENTAL CONTROL OF VARIATION IN HUMAN SMOKING BFIIAVIOR Ikscrihed here is a melhod for partitioning variation associsted with hu- man smoking behavior into several cnmponents: additive and nonadditive lrnrlK, prenaid environmental, podna/al familial environmenlal, and post- a natal eatrafamilial environmcnlal. Additive genetic and residual correlations between slrwrses, and arrrclalium between parental additive genetic effect and progeny nonaJJitive genetic and environmental effect can aho be eslimaled. Variance estimates arc free of any bias that might result from such cotrsla- IMMs. Statr.aKal genetic analysis is currently being used in central and oowh- ern Sweden to study a large group of mowo=ygolic (MZ) and di:ygolic (DZ) Iwins, their spouses and adult chiMren. Stood samples frorw each subject will serve to identi/y their genetic eorsslituliott for a number of bioehemicd pWy- morphisms. Some of tbese can be considered a rinr/ 1o Aave uosne relation- ship to smoking because of their predieposptR or esacerbeftinR effects on res- piratory diseases. To be sought are associations and genetic IinkaRes belwecn biochemical marker koci and quantitative behavioral Irailt. Of interest are a wide array of robaeco-use variabkn, mo/ives for snwkinR, personality and coRnilive variahles and others associated wilh drug use and health. Whew based solely on external morphology and questiotutdre data, the MZ to DZ artd D7, to MZ misclassiAcalion rate was 0% and 6.15% respectively, as show. by biochemical :rgosily determinations. The estimated worspaternNy )atio of lathers to their supposedly biological children is 0.28%. For 21 muker loci, gene frequency estimales indicate that the sample of Iwins and their relalives is fairly representative of the Swedish population. All loci were in Ilardy-Weiwe berg('astk equilibrium. and there was no evidence of associative mating /or biochemical Irails. Regarding whether they have ever had a smoking h.bit, the MZ twins are significantly more concordant than the DZ lsnin., Results support the hypothesis that there is a certain amount of genetic control over variation in smoking behavior. Further verification of this will depend on the outcome of more detailed analyses based ow the methods presented iw IAh report. CrMmprArr,-n. w. en d. Acro Genrrk.r MrJicar rr Grmelldorf.r 2B()) :17)-195, 1979. Frwn the InstNute for Betu.ioral Oennie., the Department of Envirowmental. Fopulation and Organismic SioloRy. and the Deputroent of rsrcholop. Uai. versity of ('olorado, eoulder; Departmewl of Ewvirorywental Hygiene, TDe Karolanka Iretitwe, and Department of EnvironmesNal Hy=iene, Swedish National Environmental Protectiwt board, Slockholn:; The Boys Town Inati- tute for ('ommunicaions Disorders in Childres, Crei6hton University Medical Schod, Omaha; and the Depanment of Staistics, Colwado State U.iversitr, Fort Collins. SPOUSE SIMILARITY IN AMERICAN AND SWEDISH COUPI ES Two s.rnpks of coupks--wrc American ind one Swedish-were eaamincd lor phenotypic identity. The American couples were white residenu o/ wbw- ban I)enver. Cokxado, an rea that was chosen because it appeared to match the Swedish suburban .reas .d %torktsolm and pd,cborg in population sire, ethnicity, and educational aml occupational slatislics Overall, a very similar pattern o1 hrmwsRamy was frNrnJ in the Iwo casupk sampks for physical, social, 84 1 MS

i and behavioral variabin. Included in this paper oe dncriptioas of assortmeet for frequently studied variabks such as social clan, cojnilive abildy, and personalrtr. as weH as odher variabks such as measures of family environment, leisure habils, and use of common substances such as tobacco arid alcohol. lhe two samples permitted some erou-aatioaal comparisons on thc eatent of assortment for Ihcse variables. Ahio, the data presented here indieatrd that the ohaerved sirnilwity of .pousn for sAOat variables was due to initial au«tmer rather than so convergence of phenotypes afler initial contact. EacelMions were t\at spouses did appear to eonverya for highly plastic variabks sucb as akohd consurnp/ion and rnou.t of socid adivity. Price, R. A. and Vandenberg, S. O. (C.wwparlrr, 0. W.) srA.vior Grnrrki 10(1) :39-71, 1990. MA., arpp..tr National InwitMt of MeNal Health. From the Department of Oenetka, Sta./ord Universitr, Stawford. ('alifornia; and the Instituwe for Itehavioral Oenetics, University of Colorado, Soulder. CHANOFS AFTER QUITTINO CIGARETTE SMOKING In this study of a large group of cigarette smoten who underwent re- pea/ed nwhiphasic health checlupa (MIIC.), 9192 subjects who persisted in the smoting habd were compared with )823 sublecls who quit between two MII('s. In all cases, smoking habits were assnsed by self-administered ques- tionnaires which covered smoling freauencr, chronic cough, eaerlional dyspnea. eaertional chest pain. other chest pain, and eaertional leg pain. Physiological measureroents that were also noted inch.ded electrocardiographic abnormalily, lotal and I-second vital eapacity, wei6ht, systolic blood preuure, serum ehoks- lerol kvel, aerwn thrcose, serum rric acid kvels, herno6lobin, and kukoclne cow+l. The prevalence of questionnaire-reported chronic cough fell markedly in tubjccts who quit a one-or-more pack/day habit. Howe.er, chest pain, short- wesa of breath and etertional kp paM showed r.o consistent improvanent in quitters compared with penisteM snwten. Weight gain was about 2-3 Rr. greater in pNters, but changes in blood pressure were small and not cow- sisteM acrow race-se: groups; nor were there consistent differences between peniaent uwoten .and quitten i. Irends In vital eapaeitl, cholesterol or prevakwce of F,CO abnormality. Quitting was associated with increase in aerwn wic acid kveY of ahout 0.2-0.3 mg/dl and relative falls in hernoglobin, kutocyle count and acruet glucose kveb, all consistent with smoker-nonsmoter differences previousy found in erou-seetiond studies. Eacept for the small increases in weight and wie acid kveM, quitting srnoting; did not appear to increase risk of coron.ry heart diaease by other mechanisma. Frlyd.n.n. G. D. and Siegelaub, A. S. Clrrulnelow 61(e):716-727, 1950. From the heparrment of Medical Methods Researeh, Kaisev-Permanente MedKSi ('are Program, ()alland, Cal. 86 THE FINNISH TWIN REGISTRY: BASELINE CHARACTERISTICS. SECIION Ill. OCCUPATIONAL AND PSYCHOSOCIAL FACTORS This t.wnograph compkles the docwneaation of a questionnaire Msdp of a twin populatiom derived frons the Firtnish Central Population Registry. The psycho-social-econornie variables, which were not covered io earlier sed:ons of this study, are reported here aod their dislribwiow is Bi.e.. The questionnaire contains I S questions related b oeeuptliond factors whieh elicit inlormalio. abotrt education, physical activilr N.rort, eroployeresx atalu., characteristics of wort, and income. Chanp of plaoe of raide.oe a.d work- place are aMo included. Marital status /ro.w two different wnrces-oAk1a1 registry data and questionnaire data-ia oowspared. 'ilte following he psy- chological seaks measuring different eharacleNsties ae wcd here: tatra. venion, neuroaicisrn, life utisfaction, nervousness, and type "A" behavior. Related to these are auntions on the anrount .nA quality of akep. Nunrrous tables of these parameters are preaented throuphorA this wrort. In the swnrnary, the main results are gathered together and awayen on the pairwise data with respect to sea and age are presented. Alorrp with a discussion on ,he use of such estimates in twin research, comparisons using different euwuates for heritability are carried oul. Kosteavuo, M., Langiavainio, H., Kaprio, 1., R.wrsdo, f., and Sarwa, S. Knru.nrrrrrtysrierrew J.dRdrr/. A/ 19:1-129, 1979. From the Department of Public Health Scienee, University of Hehi.tl, Ilchiati. PROSASII.ITIFS OF CONCORDANCE OF TWINS WITH RESPECT TO OENETIC MARKERS: A GENERAL FORMULATION In twin studies there has always been nt obvious need to differriwtiNs accuralely between mora:ytotic (MZ) and ditlrpolie (DZ) twins. Sina the availability of an increasing t.wwber of blood twuten has made twin :ygosity diagnosis by blood marten a snelhod of eheioe, formulas for the cakulnios of the probability of erono:r6ositr and other closely related quaaities haw been developed by many inveslipton. Iw this paper, the forrrwlas .eede/ 1n the determination of monozygosity iw twin pain wiwg penetic martcn Ka derived and presented. A general formula for the eakulNion of eowco./a.ce probabifities independent of tene frequencies and aflek ownber is derived and presented in a form that males computer propamming possible. In aQd'Nion, tables of formulas are prescnted`shoutd manual calculation be more eow- venient in special eases. Ilo1h scts of formulas are presented and derived In a totally Ceneral fwm. S.rna, S. ( R.nras./o. 1.) Arr. Grnerkw Mrdlrar rI (irmrllobci.r, 29:123-1 12, 1979. From the [)cpartmeM of Public Ilealth Science, University of Ilelsinti, Helsinti. A7

DEPENDENCY OF CONCORnAN('E PROSABILITY ON GI:NI? I:RIiQUIiN('IFS IN GIiN1:11C SYSI NMS I-OR T11E I)IAGNOSIS OF "IWIN ZYGOSITY. A(iRAPHICAI. PRESENfAl ION I:NABI.IN(i 111E RAPIn, OPTIMAL CNOICE OF GENETIC SYST EM Simple Mendelian markers are the most effective Iooh for the accurate determination of twin zygosny. Because the gene frequencies o( blood muken vaiy in different populations, however, the same markers or combinations thereof are not necessarily the rsasl e/eetive ones for diagnosing zrsosdy in every population. C;enrral formulas have previoudr been presented lor de- Iermining concordance prohabilities, reWive odda of dizygosily, and proha- bildKs of misclassifxatiow in twiw zygoaity diagnosis with the use of Mendelian markers. lhis paper deseribes the dependency of ptobabiliues of phenotypic concordance of gene heQuencies in three-allele genetic systems. The (ormerly puMished Iormulas are applied to derive a sd of paphs that enable rapd comparrson of the relative eAcctivewer of ddlerent systen s• with the dominance relationships within each genetic system taken into account. 1 his method can also be used to approsiinate forw-aliek systems or more; two- allele systems are considered specialianlances of three-alkk ones. Sarna. S and Kaprio.l. (R.wrardo. L) Arre Genitic.r MrAaar rr Grnwllofopiar 28:1)7-177, 1979. From the [kpartment of Pub/ic 11ealth Science. University of Helsinki, Ilehmki. OPTIMIZATION PROCEI)I1RES IN TWIN ZYGOSIIY DIAGNOSIS BY GENETIC MARKERS: A (:UST-EFFk:C.`IIVENESS ANALYSIS More and more genetic markers are becoming available for twio zrLositr determination, but only a relatively arull number of these markers is needed for adequate zrgositr diagnosis. A choice of markers can be made, therefore, and this paper presents a cost-e/lectivener analysis for optimizing the choice of marken to be used. In this analysis, only phenotypic identi/kation and knowledge of populatiow gene frequeneies ase required. In add'Nion, determina- tion costs a.e takew into accounl, as genetic rearker identifications are e.pensive and may often form a substantial proportion of the total study costs. Optimiza- tia+ of these costs without significant loss in genetic e/ficiency is possible as presented here. Data (rom the Finnish Twin Registry studies are used to illustrate the analysn. This paper also presents a method for determining a p:orf the minimum sample size .ecessary to achieve the desired sample accuracy in zyMer diagnosis. S.rna. S. and Kaprio, /(Ranrm.fo. I.) Arra GrnrNrr Urluar rr rrnsilloloelar 25:179-145, 1979. fn.m the I)rpannwm of PuhIK Ilealih Science. University of Nelsinki, Ilelunki USE OF MULTIPLE LOGISIIC ANALYSIS IN TWIN ZYGOSITY DIAGNOSIS Determination of twin zyrosity is needed in nsoM awin studies to epabli& which twins are lenelically identical and which are not. Blood twarker de- terminations are generally regarded as the mosl aocurale. but bei.g eapenaiw and time consuming can only be used in a series of relatively anull studies. For very large twin seria, questionnaire or iiuerview tecArdyuea have bee. found to be quite reliable and accveate, but there am always soree 1wiw- p.in that are kft unclassilkd. Iw this uudy, the ability of .lochwie nKtbpda to class- ify twin pain kfl unclassified by deterministic rnelhoda waa assessed by eo.n- parin6 the classification results by nwNivariate andysis lo the zyRosity diagnosis obtained by blood marker tesn. The results showed /ha1 multiple logistic dis- criminant analysis could classify twin pain previously left rnclaasUkd (XZ) by the delerministie method, and accuracy of elassiAeation was verified by Mood testin8. Mean imrapair diferences of k,cigM and wei6A1 of XZ pirs were in between dirygotic and monosygotie pain, bu1 the frequency of personal con- tact between XZ twins waa.ignilicantly kss thaw Ihal of either MZ or DZ pdn. Both r lassi/kation methods, whes used separately with the same auesliay, left unclaaifkd 6.5-7.6% of all respondent pain in Me Finnish Twin ReRistry, bu1 i deterministic clasibcaliow followed by logistic analysis misclassi/kd only 1.9'X of an twin pairs. Sarwa. S. and Kaprio. J. (R.wt.r.fo, I.) Hrmrr Heredity )0:71-t0, 1990. From the Depanmeal of Public Health Scieace, University of Helalaiti, I/elaieki. y . 89 ®

1 r / i 1 Active Projects Followie6 ib a Ibt of the principal investigators. or inttitutions. of projects under way or activated in the period since the previous Report, together with the respective projeet titka. Compkted projects are listed in a later section. PR1INCIRAL INV frSIICATOR OR INSI'TiVi10N IIARRY N. ANTONIADFS. t'f D.. Srw- i.w /w.•rsdraor and Anotiorr h r.- snr of IiocArndurl. Harrard 3c of hblie Itesbk. Ceaer /or Akood Rs• starJ, tloMw 1051' PII C AR('OS. D Sc-. Iro/ruor o/ MrJ.rrwr. TuIsne Unieersilr ScAoo1 0/ Med.cine. Ner Orksns (No. al Ihe Fnruown.enl.l thaeclwn Agency. W.shi.ylaa. D C.) MAR11 YN S ARNOTT IRAS(Y)). rnl U. Aruu.wr /wrll.rr.r ewI A niu.wt Irnfrssee of Siobrn. 1Le Uni.erwr of Tea. Srwen. (-ancer Cenler. M O Aa- dersow Ilospu.l and Twnor Inui/we. 11ouMOn I.FSI IF. RAFR. M O. As.oriNr Profrr- of Mrde.ur. Columbia Unirerser so, ('olkre of I'loysieism i Surreows, Ne+ Yort. W/I I tAM F SFNFI)1(T. M D.. Assis- r.nr ho/rn.r nf /rlbrrirs. Ururersily of Soabern C.Idornn School of Medi- eine. Dirision d/lemaddo{r snd Med- ied Oenelics. Cttildrew s HowiW of l w Anreks. Los Angeles. IIF.A 1. .an den 11FRO. M.D.. RrsrorcA PrJi.rrkl.w, AJjrwrr rrofruar in !lo- Moriairs. l).i.ersily of C.lifo.nia Sclsool of Public Ikslitt. Oat<laod. RICIIARO 1. BINO. M D., ho/rssor of filrhrfwe. Uwirersi/r of Soalserw ('ali- (aaia.Sclool of Medicine. los An. ~eIts; Yo.nnr Assorialr in A/awrJ/rol F'nrlwrrtinr. Ca/ifan.a InNilwe of lecAnolorr: Di.rrrw of CrJiolor>r and tnawrnrral MrJ.r/nr. /lunlinalon Mem.r..l 11atp1.I. hsaden.. Cal tflM~//l • M/\W~\ rAt~ /14 ......... •. .....r .l 1•..! M...L.r. 1 .....•... I Yn. ..J.y• Il.....d PROIECT TTTIl Ho/nan Plaekt-deri.ed rro.A faelor (PIK:F1: relalionsAir 10 Mrwaw a/hero.ckrari. Adi.aliq wlel.to/lrn .J di.llrl and crclie rWros.Y1i/1te and genetics of the respse lo ewry.re Inducers. Re1.lion- slly lo hrdrocarbon-wilrosamine syw- car Hrdrocubow melaboliunr en:yrnes and lona cancer Cituelle srwottinR in wr.nwlensi.e and Apperlensire subjects blood peswre. renin. sldouerone and calcctsolnnine responses Malisnanl transfonnair+n. rnWafteftesis snd ~o7noler sl[li.ily of cigarelle swlole eal0ensalt frac/iot•s Malignant Iransformslinn, nnrlal,ewesis and pronsWer aclivity of ciprt/lt're- taled rnalerial and the irnporlance of • Flasnu (raelwn (!•FI) 3s Ihese paraw- eltrs rulwrow.rr fwwclion tcsls in adokaeMs and tAeir fiarenls--sn epdesriologk appoacb ('Iw.kslerol inhibilion and carbon rnonot- ide and sUlerosekrusis I ipoproleins and the alerial ..11 1 Orr1. u/ nu.N.ne anJ 1vn/.dwlptrenc un n.unul and al..pa h...o...nt p/.duc. lu.n .041 t'R/N('IL•AL INVlrSMATOR OR INSi1TVI1pN 1. MARK RRAII(:IILFR, rrs D.. At.i.- r.nr Irn/r».r ol fA.rwr.rnforl. Norlheastern (Mlio Unirersilies Co/kae of Mcdicine. Roo/aoal. DAVID 1.. RUSRFF, h/.D., A»oci.re Pr../rs.nr of siolor,, nepr.wsr.y of 1ri.d..rirtl Sr irwcrs. NortlM Teaas Sra1e l/nirersitr. Denlorl. AI.RFRT CASTRO, hl D. Dkrrrnr, flor. sn..nr RrsrMlA [.IMMwr.wr; Ir..frsrnr ol ra~•J..~I and A(rllrtnr, lJwi.crsil o/ Misrni Scbol of Meditiwe. M F1a. FRANCIS C. C/IAO. M D. PM.D. Srw- lor InrtsNrNOr, Ceaer fa sloo/ Re- aarc); tlo.lo.. CHARI FS O. COCNRANP M D. Me.n- .er. Dep.ern.ep of /wsn/w.qesrwforr. Scr~' p. Clinic and Researcb Forwda• tiow, l.. loRa, cr. ALLAN C. COLLIN3, rs1.D, Assocfore t•rofr»or of IAorwwcelo~l. IwailMe for Sebarioral Oe.eliesl vai.enily of Cobrado, tlo.lder. PAUL T. COSTA. Jr. rrl D. Assocl.re holruor of rryrholorl. U/li.enily of Massactsllselts a1 tbroa. Dorcberer. RObFRT FCHT. I•11 D, trro/rewr of Awe- rninl. MicAillan S/ale l)wi.ersilr. F.asr CARI lY)N K. FRICKSON. Pr1.D.. rro frsMM 401 t•Aerwwul..rl. The Uwi.e.si/r of Teaas ('olkge of 1'lo.rnr.er. Asnriw. V. OFNF FRWIN. hl D.. Pr../essnr ..f rbn.a/a../..er: Dran. Uni.ersNr of ('olorado ScAuo1 of Mar/nacr. RtwdJe.. . IUDITII ANN FOSTfR. LM.D. A..xl- .rr rrofn.w of 1lorlansisrrf. Uni.er- silr of Georgia. Altsews. JACK W. FRANKFI. Put). M.Ji..J Rra.rrb SrrrNr. Veurans Adminis- rrrln.n. Nay /'ints. 1'1.: f/wr/a... lamps Re,innal I.borsl.ry. (kParl• mcnr of IIca11A and RAsbrle.lire Strr/ce.. 1 aa.pa. 1' 1.. 91 PROJECT TI1Ri Tile dleralion of tuanrfare crdass by wilric oaide The biochewsical awd 'hysiuloRiCal cb,.r. sclerisrip of a FraeiO .rbicb qeti/- eaRr bi.ds polycyclic aror.aic t,rdr. nrbon. Nica~y !{oad: deteaion by rd{. r FIMek1 acti.alio. a.d b{ood byp.rsoosalls. Mbi1Nr Tle wledialiow of inRaelaalorr fmWr of lisa.e Medialiow sraens is islR.~r..(af {•y disease (~e`~.M..eh~.i.: f ~Remo ao~~eak ~ .l SkAd The relalions between .nwtalR oai.n, Krsowalilr aW feelinp TM eRecrs of %rpak \rp!~ tarb.. IwoMSaide swd Ire.UneMe MR.e/Ks11~ ~rlwafa (oticilr ow endoeei.e-litt. eeRs iw .ir.rs)h of rwnR r.bbN. aad raNil klwes Rbod hr.iw wonilori/y of s.rai.ed .k.W liwe k.eb is rals Actions of nkaMe ow Isolated faatwe/ sworsc brain Iw.ol.ernewl of elark Abers Iw Iw fa diilsar Srnotiy and lung eanoet: fea.iNtilr srudr of a diasnonic lea lo identify persons at Ay\ risl

ERiNCRAL INV[-TTIGATOR OR INST1TUilON ALLAN P. FREEDMAN. M D. Asds- r.wr Irolrrsoe o/ Mrd.caw. H.hwe- nuwn Medical Colkde. rldedelN... AARON E. FREEMAN. M D, ScMnriM. La )o11. C.woet Rese.rcJ Fowd.lier. L. lollk C.1. LARS FRIRERO. M D., rrofeuo. .wd CA.f.w..w. Drrrrt.wcnr o/ Enrba.- wuwc.l N7~ we. Tl. K.roliwd. Ieslb Iwe. Sloc\soMr. K/FI 1. 1 UXF M D. Aswrl.ce rr.l.uer of Nurd.yr. Tlse K.robntt. I.stnMe. Sroclboln.. MURTON OAI 1)ST(Nl, M D. Asw.cW0 rr..l.ssr of M.Iw.n.. New Yorl Uwi• •erulf Medical Cewar. New Yrt. MK-IIAFL C. OFOKAS. MD. rwD.. rr../esse+ and rerrCAerwws. D.~« r- wawr of ~IelkMe. Uwi.erawy e/ Cdl- fornigs School of Medici.e. D.eie. TERESA GESSNF.R. rw D. Awriw Cawcer Res.rrh S.www. Neehh Re- se..c11. Inc. RorwtR hrll De.wwo, suR.lo. IACQUFS,E. GIEI FN. h/ D.. Au.a.wt Frolr,lo/. II1rrOrr of WedKal CAenusrry. Tosir.dofy .ed Nftw.r Isqilele of hiholotll. UniversNy Liege. Licse. Sel/iww. RONALD W. QILI.ETTR h/D.. Dlrer /o.. task Sclenrr Re,trch Uwir. C.n- ar Ceaet of H...ii. Uwieenily ef H..nil tl M.wo~ IlosoM+. GERAI.D /. a1.FIC11. M.D.. Cens.dr.wt M Mehnwr. Rese.rcf LtaAoraory /or ARet& Diseses. M.to Chek .ed FosuWMiow: ho/rowr of /nrnn.l M.Iklwe .wd lwrnrrwod~ff. MoPo Medical School. RocKetler. Misar. WARRFN M. 0010. M D.. rc.dr»o. of AIrdKjwr. C.rdrevMCrIN RereYcb Inaiw/c. Uwiversily of Csdiforeik S.w Frencleco. /OHN W. (iORROD. D C(' . t..crrr.r In I/opANT/e,. (-Aelaee ('ollege, (Jn..er- sny of I ondon. I undon PROJRCT TTfl1 The etect of cigarette smoking oe Ihe .Ivee+l.r clea.noe r.u Of incrt drsl prticlee i. the beet.w /e" Rodea ..d Mrw.n by eplletid «B c.l- wfe Ns tool for tYGeotentYf research M.it.o E~idasiologic.l rese.rch ew Ihe Swedish t.ie rep.uin Nicotiot eweetsol..nises .nd neu.oewdi. ctine /wscewns RioeKemicd b.sie of ediyosiliow to drowic oAwreclive ~isonery disese CireelMisy pncreNic e1.M.se 2.nd eet- lhtasw.. ie..s ryreseooF.e({o of towjugslior .ed Irrg ca.eer risl Moduleliow of avyl hydroc.rhoe \rdrosf- Lse end eoosiJe Ardralest ie arn.l lisstses .ed iw cell cvhwe by ciR.re/le swrole conde.sNe and other chemicals RioeKewiul mteRsi.w(s) o,d oa.lwe- live .nd qraMilMive eonstaleKes Of kieaof.)I!'re.e metabolism ERed of tobacco byMedods .d olrer eevMosrwen.l oonts*iw.nle on 1)aw- pboid aR RowrYy ..d rwrc(iow IldM.e tlfFetseaili.ill le ..1i0ews Iroiw loe.cce Elleat of ozone on .'a...1' Tta+r ctMs Teilodeck.l sWBks ee 'Flittdiwe N•et- idr: sed some telaed co..pwsnds lt+e inRuence Of pregnancy on /he ine- Lhule.m and esaetiue Of nicoline 92 i PRINCIPAL INVF1fi1CATOR OR INSry('(fMN 110DA A. GUIRGIS, rw.D.. Aswrire Irol.aw Of ("..nrwrrnuF ow/ F.nri...w- w,.nr.f Mr/iri.r. Universily of Cdi- lorni. College of Medicine. Irvine. HIRA L. GUR100. D.V.M. M V. Sc.. rM.D.. A u..e rr. C.w, er Rt.rrr b Snrnrisr. Drrorrwrrwr of F.ryreier.nbl TArerrr.ucs. Ros.reR Eu& Memorial Ins/i/we. RuReb. CAROLINE R. IUI.t.. M D.. Auor/.re rrolnwe of ydireKS and M.lunw. l)nivershy of Rocisee/p Sc\ool of Med- kfne. Rocdesler. N. Y. I INDA M. /IA/ 1. hs.D., Aswss wwe Insw ol (:.nnks and Nrwowkorr. Alber( Fieslein ColkW of Mcdicine of Yes`iv. Unhersiy. Ths Rroe., N.Y. MAROIT /IAMOSH. M D.. Resr.reA Assoc.ur. Drr.nwsrwr of f Ariolo[r ond IroMysirs. Geotpelo.rw Usiver- siy Schools Of Medin.e asni Dewlia ur. W.sbiaMoq D.C. PAUL HAMOS/I. M D. Assot:nr ho- In,e+ of rAl,iolodf .d Iw1.Arsits. .wI Mrlkine. Gearyeloem Uswversilf Scbds of Medici.. ..d Dewliarp. W.eisiwron. D.C. NORMAN W. HEIMSTRA. rn.D.. tio- Ir,.or of Psychology; Doeea.. Mrww. Fonor, /.oia.corF. U.iveesi/y ef Sorlh Da4a.. Venwi/lioe. CAROL 1. I/E.NRY, PR.D., Dbtrtiw. Dr- iwewr of E,/er4wewrd Oacerfutl: Mwalblo~ic.l AredMe4 I.o ie. 1~e.dta, Md. HERRERT R. /IERSCOW/TZ. rw.D.. AssutiNe /ro/n,e, o/ MwvelldeRf. OearRelo.e VervcrsAl Sctsaols of Md- kise eed De.hslrv. W.sbisq(ow. DC. JEROME 1.. /1OINACKI. tu.D.. A.ds- rowr P.alswr ol 1f1.rl.rKY..I s1/eMr,. University o1 I o.ell. I u.e11. Mes; WAYNIi 11!)1ti. 1nD. An.vonr fro- rs.w. ('awer for Mnin Nex.rsA. wiversilr of Rochesla Medir.l Cea- ht. RacAesler. N.Y. AARON IANOFP. Fn D, tro/.uw ol •odlolotr. Ile.hh Sciences ('eMer. S/Ne UsiversNy of New Yorl .1 S/e.sy Rrook. S/ogf Moot . 93 rROIECT TiTLt AHH .wd related bwwlrecbewkall.pt. ers: a wsoepi4iily /edes fw Lns u.oa Rok of genelks .wd EnlY.rornaic t1Nr. c.rbae wrcl.lolisws iw Mww.. hsy c.n. ter T6e relMioesAiE of lower nsEirao./lrsd .kse.u ie iw/wrc~ /o I~e develoF.se./ of chronic twy disese iw .drhs (lewelie diAerewen 1e wkolf.e acwahi.Mf ie DrosqAif. war/.nod.utr wr.isti rerin.ISl lung develoFswee(, weuboYsas as+d eiearelse ewrot. Elec( ef awotiy os evol.lle. .f +. s+wv.w ssEirMerl Ro.r-vohre aWw Sonle RA.viar.l oeEec(, of retatq ..d ssedMl{ de/rieMiee E..LNi.ft md d.r.perlwlsw ef r./- ~af11Af e1MiM Ms.f tl.0leMlr. Of ~- sauwert DNA daw.M iwbad hr elar icd ~ oe IM dselac.ls M ERecls of muwMored /nbdMioe of cip- teAe eswole oe iMMw.e tesEwsire.er Ciduelle "by end high deesil) IiFo• raei.. Studies of nicwine inler.cllon wish blood eeRe 1'wlher r«dies ow wNas.ion of pe• 1e.se iwbbilqw by cyueoe swrof,e (nweywnMyic assay of krnR c1eNin de- a.dnwww

PRINCIPAL INYPSi7GATOR IROI[CT ?Ti1.Z OR INST11UT1ON DAVIn A 1(111NSON, t•n D• Auia«nt rr.r/rrNw «/ R.w hrm..nr- Faa1 ltnnes- ue Srale t/nmvcrony ( ulktc of McJ• kine. L.Anwn ('ay. IFROMF KI FINFRMAN, M D. Ir..- /r.N•r .nJ (heOrmrn. /)rywrrnlrnt ../ P..rh..A.Ocr. Miw.a Sinai S.Aw,t of Mcd- kine, New Yur\. KLAVS F Kt1Fl7NfR- hsD. rr.•/rs- sor ../ 1Nrhrnnlfry to fklh..prdNr- f)r~+.INIrn1 ../ l1NMhrNn\Irr, Rush ('.~Ik~e o1 I/ea11A Scunces .nd Rutll Mc.l.cal ( .dstlt, Ra.A rtesAyanun Sl. I utds McJa.l ('enrtr• ( t•.c.yo Fmh.gcnous pro/eoly/ic entyn.es end in- h.eN.rs of Ne lung Dislribuliow end response of uninc pe- eursor upl.kc and dccarl.o.ylsliow /APt1D) «lls and ner,rotp~IAelial bod- ies /NEta) in menunaliaw lunIts l.ocd retulaiow of swrm.l and pllsologk iw.esio. ARFI LA/i IIA, f N f) . f)Mrrf.r, New (hnelk Sasis /or nko/ine respoww Y.NI Slare RtKMc/l IMIMYIe f.M NeY- rncAem.ury snd Drvg Add.clion. New Ywft. F CI INl(1N I AWRFN('F, M l). A/d.- FReds of eipreue smnlint on irnmonn- trnf Ir../rsuw o/ Mr/« I«r. Ray1/w ('ol- `lulwsl.w produclwn by Mmun MoncAi.1 ktc of MeJicine- Il.ws/on 1ymDitocHef lAM1 S C. I FF- P/t 13. A..u.r.nr rr.•/rr- tnr ../ RNr h.mi.ny. Sa.m I oun School of MeJ.clne. SI. I oun. Fflecls of lit.nds on plalekl micrulu6ule assemMy PIIIl Ir M. lF V(1FCNF, Tn D. D Sc'.. Assay of osygenateJ uernls in human rr../rssrw of (-hr..n/fcr• NwlAcsucrw Mood vcs+cls-a fcaubilay s/udy Umversily, lloslon. JAY A I FVY, M D• A/ryirrr rro- f ossiEle genelk deler/mnanls of chemical /rsuw of AIrlorlnr- Rrlr.r.h A.s«ri- earcirwgenesis .fr- C.ncer ResearcA In.tilu/e. Uni- .ersily of Cals/wma Schoal of Medi- cine, S.w Fr.ncisw. FAUI. D. I FWIS. M D, Seni.r ! rrr.wrr Elfeets of IoAaeeo .Italnida tw. cell pro- iw Nbr..p.1Ad..Ry. Ruy.l Pwip.Jsnle duclion in Ihe devebpnR Main MeJitd Schod. IlamnsersmilA Ilosp- 1e1. l onduw. IRVIN F I IFNFR. fhs D. rr../rnnc of holes ~h , ~M ~~ apeu ic .lew• Iwrhrmnrry. Umvenily of Minnesota. /s~`s mt '~y«m. SI. Pwl. PARIAN 1. 110NFTT1, t+sM D, Rc.rMrh Newroplril medialed iojury to liswes rrn/iuor of 1/e.Armiucy, tlosro. Uni• verwly School ot Mcdicine. 1laMora. HFNRY T. LYNCH. M.D., rrulrr~ .wd Chrirww. Drrrfwrrwt o/ hrres- Nre M.diriwr .w1 rsrbNr Hr.lrh- CreiaMon Universi/y Schuol of Medi- cine. (lmaba. RF(71NA1 D(1 MASnN, M D., h/ D, 1r.•/rINN .nl ( h«n«.«n- /Ir/w rnanr o/ t/n..tr.ity of ti1.A (-ol- kee uf McJ.clnc• tiall 1 ate ('xy. Sluislical-Renelk evaloalion of risl I.c- Iors iw /unt cawcer Oeaetic .nd Aiwwnerler sMudKs of rnot- i1y ssocialed cancers FReels of snalin` .nd nkMine an ~rmr1A and function o/ rwmen en•krhelol «Rs 94 PRINCIPA l. INV ETII~ATOR OR INSIITUTION HERRURT MCKFNNIS, 1r, h/D„ Pn.- tr.Nw of thrn..relrryry, Medkd (•ol- leat of Virdinia- Vir~ini. ( um1~- .eall\ University, RicMnowd. HANS MFIER- D V M- Srwinr Sfs Sti. cnrur, Tl,e /aclsow Laborabry. aw Harbor, Me. /. WISIFR MFI(:S• M D, CNww.I rro. /ilw o/ i~lIJrN.1.4..RV: l)Mr.f.w- f..w- IM.IMNf ( rArH Frl ir.tlWll..~r (/oct Yale Univer.i/y Scf•ool of fNeJiciwe, New Ilane, Con.. MICROS1OlOO1CAL ASSOCIATES, INC, fks4sd., Md. t l. ANDREW MITCHELL, Pw.D, Asdr- bwf rrolessnr of Awetomy. Wayne SIMe Ulwvewly Sclool of Medicine. Delroil. FERID MURAD. M.D.. Pss.D., Dirrctoe, D'wisioe of Cfiwkd tAe.n..ref..df. Deprfnsrwr of Infnn.t hldirine, Uni- vetai(r of Virgl.ia School of Mediciwe. Ctw1o1/eniRe. lAY A. NAnF.I, M.D„ he%a.w e/ Medklwe, rhywdory .w/ R.Nolory, Cardiov.sculat Rese.rcA Inslilule, Uwi- .er.ilr of Califorrli. 3" Francisco. DONAI D 1. NFI SON. Pn D- A.Nsa.r rro%a.ar ../ ('hr«tJff.l, C1art Uwive(. sily. Worcester. Mar. FRANZ OPSCI/, 1'R D, hn/r».w of rhrw.refoRy: Ne.d- Secfiow aw lio- demlr.l rA.rnt.rp/pey, U.ivenily of Mailt~ Maiws, WeM tkewa.y REVP-RLY rAIUFN• Ist D.- f.w.a Rr- se.rrA SrMwflrf Y- Rae.eR puft Me- raoria/1nslNu/q RuRa1o. f+ROl2Ci TM.tt Cooperative s1eHlies aimed N the develof seenl of ilenwnossays tor wicolins e.d wicali.e wre(aboliles F.ctorn cowlyd/ity Ifse 1lssue kvelc d wico/ioe and relNed compouwds Trawspl.cewld eRec7e of wihorocw.. Porwd. I/.b.iliaa of che/nkal (hwRl carclw. 1:new by wy1rRoS./iwowe derivaives: Wro .wd in ritro studies Revk. of lung eancer In ('onncqkw, 1911 pescM S^ale I/IRalaliow carcinoReeesis s/udies in swk.e Sland.rdiraliow of assays to /nesure vyl hydrocarbow bydrosyUac in fwm.. IiuMtf C°Ratar.lis'e study .ilt, Dr. lay 1.evF (U.iveesily of Cdiforwi. San Fra.chc. Medical Center) des~' wed /o de/erwi.e //re te/K(ic coMrui e( aewolroric vM>tm espessiew in hybrid Nc.iws of mloe IwIFLMMiew ud werine hwdlo, i. Ib rM MeeRwrw of rlitrk o.ide acli.allow of tirataylNe cyclase INeede.iros of eir.ey Ay'erirri1.611kF Speeuo.eqk r.dies of the ifwer.elfoo of elwli.eeSk lisawda with wieaWe n- !n'Mter Moleiws TKe rak of diAydrodiol dcRydroanese i. 14e conlrol of re.clive rAll swel.lsa Ules cernf.rcd b IhM of kIrttsM were eelewsi.el, Mudied e.tywws dtr/elk Mace'e/bi1Ny to Sladder canpr 95 •

. PR1NCIrA l. IN V G?nG ATOR OR INSnnfilON DFNNIS R rFT1 RSFN, rN D. Aahrtwr rrn/r.ur ../ rArrnu.r.d..cy. I Im.ersNy ol ('ulurado Schoal ol Pharmacy, Roukkr. CARI W rIFRCF., M D.• M D• rro/n- rw of r.uAolocr and AfirroA«d..f1-/ar. w.rwoloAl. Wasarnpow UniversilF School o/ Medicinc: Ihrr.ror ../ raA- olodr and l.h.rlrnrr MrlNlnr Tte IcsernA lloyNal of SI I.oYlt, SI. i.oas. 11 ARI RANTASAI O, M.D. rro/rsrer awd CA.Mw..m nrr.erwrrwr of a.Nc IfrdtA Sr.rncr, Um.eteity of /lcluwhi, Ilelsinli, I inland. RONAt D F.. RASMVSSFN. 11t D, Ar- .o. i..rr Al/rncr rrn/ru.r i. Cawunr- wry on/ I w.irawmrnral MrLrMr. l)wi- vcrsitJ o1 Cddo.nia ColteAe of MeN• tlnt.llvint. 1'1lFF.N RFMOI D O'ItONNF11., MD. rrrnrrpd Rrre.r.6 AN.MM/r. Ilerr.rd McJ.cal Sch.rd. ln.nnerr.r. ('emer /or Slood Rese.rch. Sosron. IIFReFRT Y. RFYN(H DC. M D, rrn. /r.rw o/ Mrd.rrnr: Hrwd. rr/wnw.rr Serr.ow• Yak I/n..ersatr School of Medrawe, New N.vcn. C'onw. UNA S. RYAN. ra D., Srniar Sr7rwtbl. Ihp.nicdaw Cancer Researe\ 1w,t1- hqe• Miawi; Resr.rcA Aur.ri.tr r.o- /rrrnr o/ Mrlicine, llniversN of Minwi School of Medkine. lirwl, F1a. • V.•RAMA SASTRY. DSc.. MD• rre/rrror of rArrn.rnl. ~. Vandcr- •iR University Scbol Of Mediciwe• NaRvilk. Teww. OP.RAt.D SHKI.AR, DDS. CA.rlrr A. lr.rlrtt Professor of ad r.rAologr: Hr.I. DrMrrw.rwr N Orel MelrrJwr and Or.l r.rAologf. 11.natd Sdool of DtMal Mediciwe. SoMow. NANO('// SI (/R• h. D. Arrnelirr r.o- /rrc.r ../ Nr..wn Crwrrvr. SecUer [clwwA of Mrdu.ne. Iel Av.v l/m veully. Irl Aver• Itrsel. tRO1tCT TTTIl In/hanccs of RenrNfpe, ses .nd chronic ciAarctte sm..linA on Raol.nc and al- coiol w.tuholrsm in wrice RioloAr of soppessor T cells p.+i/cenlelogicsl studies of the Finnish T.iw Regiwy Apwdk rodr of DNA rq.ir in Inbted Mrair of wwice ConelMkw of cellular and liochemKal evenls in Lrer of wrke ecpo+ed Io ciprale smote or l.ry to.rc chem- kale . 1'urilkation ud functional .n.lysis of elas/se Irorn suinea pt w.acroplases ~roncAo.l.eolar lavage Ruid in pulmonsr>r earcinorna. secretory eomponenl o(im- wrurwgk+buliw A e. • wr.rlLer o/ neo- plauic growth ResPirMory sccretions iw'ulrnon.ry e.r. emorma: .ecre/or/ eornpnncnl of irn• twuwo~1o~u~m-A as an early marlcr of epitheli.l Ilslwrc(iow INer.eliows of Rorwrones with cells of 1Ne rylnwnary vascdat wall IwA.cwce of .koaine on H.e teksc of .eclykholine iw Ihe human placenu .nd ile i/.oKalioM ow the Ictal IrowiK Oral carciwoAewesir, vitamin A.nd tet- inoide T/.e Y.e of tQeciwe .MihOdeee 10 mrNN/0r the fwm.uon and removal d henco- LMrrcne .JJacts /rurn DNA of J.m- a1rJ human eells Iw rieo and in vitro 96 rRIN('IFA /. IN V FSi1CATOR OR INSni1nMN 1 OUIS A SOI OFF. M O., sl.,~Ar rp trrf /)6rintrl.Ar/ Sr.rirr rr~n~; rr..Jreur. ../ Afr/iriwr; Dirtrror. Rr. K~*uI IkraJNA A"porr. Te"'~ Uni delp6ia. ~~~ Cewler. Kd.. rRO/ECi T1TLt I ecilhiwrchoks/erd acfl /raweterase aAee tNina T1M ' O1 11Y A. SFRINOFR PHD. Ar .ir..nr rrn/.rr.. ol rrrltolotl. H.r. .Md /1/CJitd ScAod• soya. T1IOMAS P SIOSS 466ddowe erewl~i.Koo.~ ~~ ~ Mwilodie. . p(' M Oy CAlt/, MrLc.f ()wryit~do~ r IJwN, Mawcwsrtls (kweral llo, ~~. I.YNN M F ~r ~ ~f of Ib F./-" . TM1SS10. M.D. Ar~~ rro/.r.or and Atra.4rr C•A~~w HeaUA~nr o/ rdi.rrfrr. Ar' Scieectl Cewter. T.csow. ra Effp e of ~K. ses .nd diseass ow /he int bUaraw lyry i ANDRF.W M• TOMF.TSKO, ~ D~ rrra Darrra. o/ Rew~rA, L'Ntow ~ti Lt1., ReaieslpN.Y. JAMES ~TRAVIS, M.D ho/rgor ~«t~ Mhewe~ ~lwive(epf o1 1?': ~A: u ~N~ M D, .f.INwrbalr .u1 Medicd =2~a~of/. Hr. UNION CARe1DE (-OMPOR ATION, N.clear Diviriow, Oa1 RiJ~e. Teww. STEPHt.+N F. VATNER, M D w/r- oj_A/•I,ri.v H.n 1 MeOi. ScRool• New P~Iey ReFiew.l hi, wrMe Researt# C~r, SowAMro, M.a Yc/wsNU• Arrnrierr iw Afrlirinr, hler ResM ~yh.wt H i oq ld. {oyew. IRENE Y. WANO. ot/ snI Alirr hr D. .(ulrr.wt rrs /rr~r of toir aw/ C/iu(rd /wtwrawo/. wAblosr, Mtdicd Ury. .erwy ef Sowf1 Cardi.R CUwkslow. oEOROF WFINSAUtN. Mo, .bwi r~u. rr/wrow.r' Disr.w Srrtiow. A/• delphraAen 1'imtein <Nelica/ ( iwter. 1fit.• . AKF WF.NNMA1 M. 0.1 D, lr.•h.rr.~r.wr of Cdw.rr/ rArrx.Lyr .e K.ru4nr/e /wrruM~. Huddinse I/usplal. HuJ- disse. Swekw. Plobirel wkoliwe rettFlor siles hN d~~ -ftwee and Lkihilps N.. ~ ~ 01 b.wul ~aer Md aatl.Ne1 eR.r.ek.ir"h w a.YtsN wel0, iFw.eo Aevk.. .d 10.{. NW~ ~ed rdel c°eowr a..erL LlDkeeg ie.aRed. K wkelirk ow brd. c}g,. Ir11riRlalioA of .MIIIIlt1. /pwy a/ AssM live "kfosOmV e1locMarwe R1fR and tMe ~roleiwa~:.~1 eAed ol c iMo/eMase ~~ iwleraqiew yaretle snt"e ow tbi. Rrawchioal.eolar Ia.aM d brain .w snwl. ets aM w.w~ten: N~k, on ceR chema.eh~ enerwr etk.ee awd « Ilu- Iw .lttWt~~e Nicaint as iwKi6itor of 1°aLion : locdintiow t41•~w /o.- sleo and edaraqeriraliow ~Mb~ory rauYlar "~ IIK tardw m1Oicaio..

PRINCIIAL INVt3TiCATOR OR IN5111111WN IAMES A. WILL, D V.M.. M D. ho- /tssor onI ('A..rw.on. I)rIP.rrnrtns o/ Ytrtrsnry Srvnrr. University of WiF cweirl Mediso. ALVIN WINIFRS. ru D.. Asslu.nr rro/trwr of Mriiro/ Murn+lkiotl. VeHere.s Adnrinisu.liow Medic.l Cc1b- rer. Rel !•ieeti F1e. RRUCE A. WODA. M n, Rturrlt Ar- s..n.rt, City o/ H.+pe Nerbn.l Me1- ic.l Cenu., t)tr.rre, C'il. OEOROE WO/ F. D hHr . rro/ssar o/ rAfswl..f.rd ('Aenrrrrrr. Dtp.rnwnr of Nrr.erww .nI Food Scwnrt. Mew- cllusens Laautc of Tecbndeft. C.srr- brd.e. FRO)E(T TITLE Morphololic and func/NN1.I c.NICL/NMt t>( ibe APUD cells of Thc lunt ERects of sn.oking on Urc inbereM iiHer- /erow kvels iw conud .wa cenccr P.- (itM.:. pild.lwly CeR wrf.ce wren.M.nes. iwtgr.l -ern• lr.nt Oratins .sd cf s. inJ o1J r.~.: /~rt1 froaw: ( I r rounf 12) Moag eweow rN IynpA wnes VMeiwh A end Rlfco}racin sr^thcus iw resNrN«f eMrbcl.wn: .rIery 6e1ec- losyl uenmfer.a, a possiWe cnly mert- er esyinc fot pledder c.ncer 98 i Completed Projects Following is a liu of the princip.l iavestifslors, or i.qyrlb..,~ d projocts that have been compleled prior lo tlle period covered i. Ihis Report. Several of the Individuals named are deceated. The lilks and aWi.- tbm lisled are Ibo.e is eQen at the time 1he work .rr co(ople/ed. L6O O. AROOfl, rN D. rro%ssot of diocAenrisury on/ tio/w KtwrcA. Ces• let lor er.in Resc.rck Tbe University of Rocliesler Medical Cew(er, Rodes- Ier, N. V. MARI() D A('ErO, rwD, Associ.tt rro,irso. o, rA.rwroroforl'. Medical Colk{e e/ Vr~ VirRiwie Ca~w.w.. .~edr` Ur.enwy. Rita.wN. CLARI:NCE M. AORESS, M D., Asw- (we Clinird rro/r»o. .r/ Mtl+tuw. University 01 C.Iilot.i. Medical Ce.- ur, Los Anglek.. ANTHONY A. ALRANFSE, h(.D. Di- retror of Lobor.rorfes. TAe llwks Re- l+.bi/Na/i0. CeMer, Wlwe r1ai.S N.Y. ANTIIONY P. AMAROSE, hr.D. I.- srtrrrw M Oluerrks sw/ c)wtrof.sr. Ti. Alb..r Medical Colkge of Uwiow Universirr. Alb.wy. N.Y. E. T. ANOELAKOS, MD. hr.D., rro- /rssor of rAlskdaty. lloMei U.iveteily Ydod of Melid.e, t1o.1o.. D. MURRAY ANOEVINF, MD.. Ud- .ersilr a/ Wi.eewel. 3Aw1 d Me~4 eiwe. MaJi.e.. ALAN K. ARMITAOl!, Pu.D. Rex..cA Dirtaoe. Herkto. L.baMwin Er- rore. Haroplq Nortb Yerk.liro, P.ae- l.wd. DUMINOO M. AVIADO. M.D., rro/t.- sw of tA.rrw.rofotr. University of rennsylvani• Scbol of MeJicau, PAiI- sJetpM.. STl?rHEN M. AYRP.S, M D.. Dktcrr. Crlio>cJsw.+..ry [."ra.y, SeiM Viwcewr's Ilospilal, New York. O6CAR 1. 11A1.C11UM, rw D., N.sNwts /ra/nsnr o/ Helsciwr, l/.i.ereiry of Soulherw Celifurnie Sebool of Medi- einc, Los Anaeks. FRF.bERIK S. IIANO. M.D., rro%ssrr o+I C11.irn..w. Dt'.renrenr of rrb- F_(.Ioty. Tl+e lotrwe Hoaiwr Uni.eedry Scl,ooi ol Hygliewe ..J Public Hed1k l1.Nh.ore. A. CI.IFFORD IIAROFR, M D., Roler( NOwrr r/NOrr rroJiuw of IApl.l- otl. ,Iar.rA Melic.l Sc6ool, Ro.N.. tIRODA A. BARNE3, M.D. hr.D, h.• /essoe (A//Nou) of Physiology. Cab r.do S1Me Uwiversiry, For( CoRiee. FREDERICK W. tlARNES. 1., MD, Asoclrt ho/tssor of tblkine. TIM 10l.. Hootise University Scttod el Mdicia. R.Aiwrae. T. C. l1ARNES, DSe, RrsertA Seir. i, Yhil.delRRi. S(w HoeEitel, Hi.• CARL O. tlECKER M.D, Aredre Ias/tuo. o/ rorAojots, Cas>eR U.i• va.iry Mdicd CoYeRe. Ne. YetL. R. PREDERICK RECKL'R. lhr.D. Aeer ti.q lro/iuor of An.rawt .c/ Dtih Wr. Lolvw.rerY o/ %riaeb/ Sc(twn. DrLs Uaivenilf Medical Cewr, Do- b.q, N. C. RALPH S. rECKER, h.D.. MJerr !r( CtmeA.wr(wl. University ef Ne.*Lq tlENIAMIN RFLL, M.D.. Dirtrtr E+wx• kns. Nwwr.wi.•e AtMt Srrly, Ve(a- .ws Adnsi.irrNio. OwF.rieM Cflde. Rowow. SAMUEL RELIET, M.D" D risiow o/ Cr/iefoe/. (lewerd //oqi(a1, Kr1.41phi.. l1ARUl RENACERRAP, MD. /.1tmr rr esw..wl Ch.Mw..w, Dq.rasawoJ ~~t f. H.n.r~ Mdical 3ctrod. 1O11N A. RF.VAN, M D., P•ro%r.a o/ ril.rwwrorotyU.i.etei,f o1 CdN«- .i. Scl.oo) of MNici.e, Los Aryeke. 99

et/p11DEV •IIAOAT. PMD. rro/tsaor ROeERT E. BROOKS. rND, A»nciar el rAfs/•'Grlf. Saint I ouis t/wi.ersiy r.olrnwr of r./AoI..Jy. University of Schoul of Mcdicine, SI. Louis Ortpw Medical Sthod, Portland. CFSARF. eIANCIFIORI, M D, Dlr/riow BARBARA •. BROWN. rw D.- Chit/, ol C.krr RrrarcA, Uw..eraNT of Earrri.wtwal rqcAie/ry, Veltrens Ad- rer.Lu, rerusi.. 11a1/. wiwislraiow NoyiNel, Seprlvedw, Cal. HYLAN A eK'KFRMAN. M D., Au/w- RAYMOND R. BROWN. rw D. P.o/r.- onr rrolriror of MrLt.nr, .wd AL- sar e/ Cliwke/ Owrolop. University of VAN L. sARACII. M l).. Cew..hwr Wiseowsiw Mediul School. M.diso.. iw Mtbrinr, Cdumhie University Ce/- kge o/ rhysici.ns & Surgeow.• Oo1d- J06EP RROlfK, PMD. rro/nro..wl wNer Mew.orid Iloyile1, New Yort. CANrw..w, Drprfwwwr o~rsyrAologl. l.eblei Uni.ersily. IIe1Me w,, re s1O•RESFARCI/ CONSULTANTS. 1NC.. C.wMiye. Mese. SUd DUCKINOHAM, M D_ Asduaw rreJrwr e/ rrll.rrks. Coluwsia t1wi- s10 RESFARCII INS7iTUM INC, esnilf Colkp of rft.icl.ws A Sw- CuwMid~e. Mau. Newq New Yorl. FRED OVOCK, hr D. Asseci.rr Ctr A• SONIA NUIST, M.D.. Assoclue rrf trr RrvrcA ScirwNsf. Ai.dogk./ Su- l!sser o/ Mt/iriwr awd rAy.lnloel, r/ow, RosweR re.k Mew.ori.l Iwslilawe, vni.asity of O.epw Health Sciewtcs Spiq.dk. N. Y. Cewler. Por11.wA. OUENTIIFR ROOFN, M O, Anntbrr BENJAMIN BURROWS. M D.. Asao- cWt rco/ruor of Mrliciwr. University rro/nror of AhlKiwr: Armr.wr Orft- ot Chicego. Chkago. for. (irntrr ChwKr Rrsrrch ('rw/rr. Temple University Ikeh\ Sciewns Cew• e. M. Rt1iT. M D. Chief r.rAotoJlsf, 1er, PhdadelEhi. . 10. Angeks Cerwy Oeneral 1lospild, IIFRMAN V SOFNIO. PN D.. Nrel, I os Angeles. Drprrmrwro/CArMlury ewllwrhrwr- RICHARD U. BYERRUM. hrD, he- bfry. Spiwdklop Rese.rcJ. CtMer, Les. /rs.or of CArwd+hy. Michigan SINe Iwwow, Ky. Uwi.ers.lr. E.M Lwwsiwg. IAMFS P. DONNER. ru D. Pro/rssnr SISTER M. EMILY CAHII.t., Ps/ D. e/ Rie1eFT, C.liforwie /wMNswe of CAdrwsow. Dtprnwtwr of Chrwsis/rJ, 1`ee\wolosr. Pes.dew.. Regis College. Westow, Meu. WALTER M. BOOKER. PwD. r.o/rs- RRUCE F. CAMERON. M.D., hr.D. see ewd Nred. D.~..fwrrwf of rArwa Ho.rard Ile Insliule. Uni.asM7 telots. Howerd UwhetsMT. weshiq. o/ M~ Sc~ d Mediciwe, Mi.w1 1owL D.C. Fle. FRANC01S M. SOOYSF, hM.D. Srw1o. ELROY T. CANTRELI., Plr D.. CA.1r- Jw.ru(prnr. Micheel Rerse Rese.sch Room. DrprMStM o/ rhrwwolotf. ForwdNiow. Chicago. Tesws College of OsleorNhic Medicine, Norlh Tea.s Slwle U/r.ersily, Denlow. RAYMOND ROSSE. Tw.D.. Assoc/.n Mr11./ /AM H. CARNES. M D. Uwhrer- Dl.rtrr, Nocwunrr A[iwg S/rlt. Vtl• w/r o/ Ula\ College of Medicine. SaN er.ws Ad/wiwislnliow UwNliewl Cliwie. I eke City. /w1ow. TOM O. SOWFRY, rwD.. Rew.rrh MARCUS N. CARROI.I., Ir.. PMD. rroltnor. rrr/.c/Ir Rrsldwe lifle.e• Chirl. Dirlaiow of PA.nw.cofofy. 71e No.lh Cuoliw. Srue College. lroolJak HosEilel Cenler, SroollfR 'I~h N Y. (•/rq/Rt Y1 •r1N1tMAN t,etl A. WI/IIAM AIVIN ('ARTI;R, MD. • MMY/I /...1....~ rf Y.Ir..u W.sfar s..r....w. P.../r...w ol 116rJu.nt en/ %/e.e IL....M/ Ll.../ .f N.e..w, Ilr L.bw. /1..P1ons Ilwi lltuor efw/y 1al..ul u/ Ned..rnt, S.IUwWrt. I 1 EOPOI D R. CFRFCFDQ rH.D., rre. It••••r .•l MwrAt.wuay and N.u.iriow, t/mrersMf of herlo Rico School o/ Mediane. Sw. )wew. JACK CIIALON, M D., Auocl.rr rro%r- sor of AwruArnioloty, New Ywk U.i- •ersily Medicel Cenler, New Yorl. C/It1.DRFN'S HOSrITAL OF 1.05 AN- (iE1.L•S, 1.06 Awrks. SANFORD CIIOD()SN, M D., Asdpewr Pro/ns.w of Mr/ici.r, T./ls Uni.er. .i1L ScMrd of Mediciwe, roqow, NAITP.R M. CIIOrRA, rwD., rre%s. ro. o/ CArw.iurl, Netth C.woliwe As. ric./hwel and Tahwlcd Srwk Uwivet- Wf. (keewsboro. N'111.1AM O. CI.ARK. Pu D. DlrrcKr, rsf'ji/PArwrrotodT RrarrcA /rA..w- r.Mp. Vtlet.ws AdmwlsrrMio. /loyiW, sea.l.ede. Cr. HANS T. CI.ARKE. D.Se, rrolr.ror el •iothe.winry, ColrwsAi. Uwi.ersilf College of nlriiciw A SwSeowti New YorL. JAY D. COPFMAN, M D., Sred.w Nrel, rerirArrel Y.rcw/.r Dry.rww.wr. University Noyiul, RoeN.. ALI FN R. COIIEN, M.D. Pw D., As- i xirr ho/rasor of Mdklwr. CAir/, trlw/ewer~ SfeNow, Te ~ Uwi.enYT &iraoes CeMer. P\ilde DANIFL COHEN, D.V.M. M.P.H_ Aa- aWav Prolrssor o/ Yrrtr!wry EN. trwdulory owJ Irllic NrdsA, Uwi.ef- siy of 'ewwsyl•ewiw School af Ve1er- Mwr Mediciwe, Phi1.ddNiw. JULItJS H. COMROF., 1.., M.D., DAra for. Cr/forestabv RrrrrA fwairwr, University of California Medical Cew- tcr, Siw Frencisco. DFAN M. CONNORS, M D. Assoclwr Datcro., Drp.rrrorwr of f nAerorwJ Mrlicba, St. Mary's Noyilal, Meliwrl, Wis. PNII IP COOPFR. M D., Cliwkel J're- lraw. of Swrgrry ow/ Dirrcror. Sartl- fel /..AorN.rry of ('rll.d.r rhrae%t1. ARrtrl Finutin ('olltge of Mediciwe of YtsMra Umrersilr: ('A..l, SwtKol Serre t. Veteran, Adminislr.uon Hw- pilel, l1ie s/ons, N V. JOHN E. CRAIOIIEAD, M.D. rre%r +.+ of rerAolo~I. University ol Vp- woel College o, Mt.daiwq RwYwg lew. ROBERT L. CRAIN. PM.D., Aaduw Hrolrasor of Jociolop. Ur.e.d1y d Ch/CNo. Chicap. IRVINO P. CRAWFORD. M Q. Pre%.. sw .w/ Clybwe.w. Dvr.rrwwwf o/ M4 e*uNolerr. Ud.eni,>r wf lo..a Co.ep e/ Mediciwe, Iowa City. T. TIMOTHY CROCKER, M.D., rt./r.. ~~ C eMc4.r. UM.ersh) of Ce1t- atlqt o/ Mediciwe. /rvir. CARROLI. E. CROSS. M D, A.sorire holrfsor o/ Mrfkiwr ow1 Nrwrw rAls/obsr: Dirrcror. Srctlow y rd- M@Omf M.Iklwr. University w( CN4 farsii. School o/ Medicine. Davis. CECIL E. CRO63, RrsrrcA Dq..yws.wf, SI. JosefY HosEiul, wrh.t, Csd. DAVID W. CRUMPACKER, Pn.O., rh.. rsror .wf CAwirw.ew Der.rusr/y o/ .Orew 4, rw.Np .1 C.Mni~, ALBERT DAMON. M.D, lh/.D. Lrc. frw ow AwrboP.fofT: Re.r.nA Aa..- c/w M AIr/k.l A/yMs~oI~ ho1/ Mwews, Hwr.wrl U-wi.~shhe- C.w4iye, M.a. THOMAS R. DAM/RER, M.D' A.eeeY. tro/rr.er of Mr/kiwr, Iloslon Ud..f- .iy Schowl of Medicir, Roao.. R. F. DAWSON, lM D./'r%.ax./ lre. rs1. Co1wwM. Uwiwsilf, Ne. Yert. JOHN P. DF.LANEY, M.D., hr.D, Ar aorlar ho/ruw of Sr.tnf. Uwisr.pT of Miwwesola, Miwweyoii.. ANDRFW S. D/RNER, M D., E.re- wirr, rsycAsResrsmicA, Tb Ap Ces~ /er of New P.rylewq, Iwe., Doslow. EDWARD P..pOMINO, M O., Prv/r.. MkMj.w,iA ti~f• Uei.ersNY wf RA/ PII 1. DORFMAN, rn D, Dkrcrw .4 1 oA.~wr.rirs, Worctsler FoundNlow /or F.oe//menul Biology. ShrewsMsry. Mess. 110 ' /nr

11 FRFD D(1WN1'Y- hrl). Arriunnt rr.r/r.s.o of t•hrri.d.Ky. Ilnicrrur ot Te.us I/rallb ScKntt ('eMer al 1)>~la.; I)rrr. e.w, Ca.Jn o..n. r/n. Rr.rurr h. ('ar • du.prlnwM+ary Inawwe. MethoJru Hos- yiul d Dallas, 1)a11as. lAMFS 1 DYAR, PsrD. Aseiuawr rro- /rssor of Riolee/. Bcllarnune Colkge, 1 oasvdk. Ky. RICHARD 11. FARI F, M D. Chir/, t wlrannNry iw. ru.w I aMrann.: As- uuanr rr..frrwr of Mrdltiwr. Oniver- ury of Cacalo. Caica.o. /O11N W FCKSTIIN MD. Asduawr ho/rswr of fwrrrwaj Mrlrrlwr, SraMe University of lowa College of MeN- ciwe. low. CMy. BERTRAM FICIIFI., DDS. Directer, iwsritrrr of Sro«wtn1..(nd R/Karrh, Science Resoorces / ound.Nioo. W.Ner- toww, Mass HYMAN FNOFI SFRO, M D. Arrrwd- ine rhvrki.n. ('edars of Lesawor Hoe- pird. t os Aqe{es. CARLTt)N K. FRICKSON. MD. As- soci..e rro/rrsor o/ rharmarel..llt.a/ Tnekolotr- The University of K.wsw School of tAurrnacy, l.awrence. /IFNRY I FSeF.R. hsD, R.rrrtA /ws- wsrwo/ogiM, Mason Research lwsriaNe, Worcester. Mass. WALTER R. F.SSMAN- M D.. hr D.. rro/rssr of Issrhdoly and r(otArna Isny, Qoeer College of Ire Ciry Ud- •ersiy of New Yorl. FhrsAiwg. JOHN R. ESTERLY, M.D. AssoeWe trofrlwr of Pathology. UahersNy of CNcaRo Mufer School of MedieMe, Clricaf/e. HUGH F.. EVANS. M D. D(rrnar. Dr- F.r(nnsrnt of rrLalrw s, /cri.` 11os'i- lal awd Medical Ceoler of Sroollyw, /rooklyq N. Y. IIANS 1 EYSFN('K, hrD. DSc. rro- lrsurr of h)rh..folt. Insliartt of rsy- ehiauy. University of I.ondow, Loo- Iow. IIANS I FAt K. I'w D. Alfrnrr A.rotL ata Ir.,frst.w .d rarh..faer. Ilniarurt of S.arllAtrn / .ldurma ScM.ol of Meda.nt. I os Angtlcs DANA 1-. FARNSWORTH. M D, Hrry K. O6rrr rralrlMM of H/Rvwr and Dirrrl.w. University Ilt.drb Sr.rrrrr, Harvard Universiey, Carnhridhe. Maa. GAD FFINSTFIN, he D S.win. ttrr- twn in lla.rh.w.isery, TAe (k.Mee S. Wise Cenler of Lile Sciences, lel Aviv UniversiNy. Tel Aviv. hrnel. FRANK C. FERGUSON. 1e.- M D.. Chawrw.en, Dr/orrnrrnt of f Aa.nw.d- eyt. Tk Albany Medical ('dkSe of llsiiaw UniversiNy, Albany. N.Y. THFODORP. N. FINI F.Y. M D, Dirrc- ror. ral.nnw.rr Rrsrar.h I ah.var.ry. MowM 71" Iloyied. San Francisco. WIl.l1AM (. FISHRFIN. M D, Chief ;L E~idrnrb/eRy. Chicago tkutd of .bh, Clicato. EDWIN R. FISHF.R. M D. Direrrr o/ l.rlerarorlre. SAadyside Hospral; Iro- /rssor of hrhohKy. Universiry of hlrsbweb ScKool o/ Medreine. /iua- lanoN• RUSSFI L 3 FISHF.R, M D. University of Marylaad School of Medicine. aal- liwsore. W11.t IAM H. FISHMAN. hr 1) . r.ed- Irwr,l.a Jolla Caoca Researc\ Forwda- lioq La Jolla. C.1. 9 L. FRF.FDI ANDFR. M 1). Directar of Cawter RrwercA. Moonr 7inn Hos- pr1d and Medical Cenrcr, Saw Fran- CiKo. FREDERIC A. FRENCH. A s.. Dirrc- for of Cancer CArMSorhrrqt Rrsearch, Mount 2iota /bqild ar.d Medical Carer, Saw Fra.eisco. JACK FREUND. M D, AssbuM he- /`sur of rharwwrotoRf. Medical Col- kle of Virginia. RicVwond. Oll SERT If. FRIFDFt.I.. 1.( D.. Chief ol rNhofu;r. S(- Vincew( IIosFNd, Wacesur, Mass. GARY D. FRIFDMAN, M1), A.uuent DireNr. Drprtwrrw/ of Mrdic a/ Mrtb- oh Rrsrrrh Kaiser Forwdaliow Re- eearcA I.slilwe, Oakland. Cal. H. HUGH FUDENSI:RO. M 0. rro%s- aw of MrlirMr University of ('ali- /ornia Medical Center. San Feawcisco; Iruln..w of rortni.d..tr anI /ywn.r- wnlufr, Universily of Cali/r-.nu. Ser1e- ky. 102 i ARl1IUR FUNST, rsrD.- Dirrctor. In•' srurrr of C'hrn.hef r.ol.yty. Umvtrsily of Saw F'ranc.sco, S.w Francisco. MURRAY 11. OARDNFR, M.D.. Auo- rwnr I'ro/nsor of Pah.d..Rl. Uaiver- sNr of Snw6erw Cdilorwia School o( Medicine. Los Angeks. GFORGF O. GEY. M D., Dirrctr, Fiw- wry H..M rll ('an. rr Rtsrarc A/.Jorr. r.My; AfUKNUr h..Jrsu.r of SrrCrry. T!e l..hns tloptins Uoiversily Sclrool o/ MeJicurc. RdUmore. 10SEFH 1. GUARNERI. hr D, Atuw/- Nf M.cr..lwlorur DMrltor. A/icr.N. istoef l aAoratorira, Long Idand lew. i.h-lblhide Medical CesNer, Qree.e Hospaal Cerner ARSliaion, law.iq, N.Y- FRANK F. GUTHRrE, hr D., he/ea- sr of Ent..wmolo~l. No.IK C.rd'w S/tl. CoRese. Ra1eiN. 11. •. HAAG. M.D- t ro/rssor e, lAr- ns.rol..e1. Medical College of VwRinik Ric~n.oed. T11/)MAS M(NX'KP M D, AsasrWe rrnfres..r of rrrrrwrirr AIrJNinr owI (' . I F. /. HADDY, M.D.. hr D., Iro'rsser ewJ Chdrwaaw. D.yomnrwr o/'Ald- l . n.«rrnuy lr.dth. Sc/on /1a11 Cul- kRe of Medurne and tkMisuy, kr.ey ('Ny. N. 1. DAVID M. ('.OI DFNSFRO, St•.D., 1611). AsYKWr hU/rsV.r n1 rNhnl- oEr. Ten.pk UwiversNy IkaNh Sci- esrces ('enMCr, MiladelNi.. o ..m llniversiry of (Hlaborna Medical C'ewter, (Nl.borna ('iry. 10SF.P11 H. HAFKENSCHtEL, MD., Dnec(or, CrJinpsda.owor~ Uwli, The l.awheww I/osFirel; Aaseciarr /M hleN- cine, University of reawsyl..ni. ScKod 01 Medicine, Mil.delrisia. PA14. (:()1 DHAt1F.R, D.D S., Assoriwrr rro/rsser o/ rraaJ..wt.daKr. Ilarvard ScMwd of DeMal Medicine. Ihrur.. I.EONIDE GOI DSTFIN, D SC., Aau. rirte rro/ru.. ../ rsjv hiary. InsliAae tur MenMal HeaMis Scae.ces, Co1kRe of Medicine a lknlisuy of New lerser. RWRers Medical Sclwrol. risealway. IRA GORI:. M D., rrn/rewr of retAof. oty. bsro. U.i.ersiMr School of Mdi- eiwe; CMr/ al [.Iw.uwl Serrke. Veleraw. AlwrwdrMb. I/orpild, Wes1 Roaberrr. Mass. GERTRUDE Y. GOTTSCHAI.L hr D. RFRNARD IIANF.S, hsD., rro/r..er of IIrdfA S. knrr, Cali/oraia S/ne U.i.er- siry. Norlhridge. RK'HARD 1. IIAVEL, M.D., Au4rwr rrofrlNM of AIrJaewr. Uu{.ersMr d California Medical Cewler, San Fnw7 cnpr. HERBERT R. HAWTHORNE, M.D. ('h.irwwn. D.~..trarwt o/ Sr ylScAeolMedi'eio~e, rildelyY., IOHN A. HAYES, M.D., Aa.ui.re rrAeloRia, Mallory lwsrilwe et h- 41106oLy. 11os1oa Ciry Hoqiul Rorlaa .lsusront ho/naur o/ riw-hrrwisrrf. Colwwbia Uwi.ersily CoRqe o/ 1'My.i- eiaa» i SwReoa New York. A. CLARK GRIFFIN. rtr D., Hraf. Drr.rrwerru .4 ri.rhrrnisN7, M. D. Anikrsow IWspild awd Twww Is,(i- 1we, University of Te.as MeJical Ceaer Ilosww. ARTHUR L. GROSS. M S., Srakr 1ie- rhneis(. SowAwtsl Research InMjr.1e, San AMOnio- Tea. MORTON 1 R CLARK W. HI?ATH. M.D !V%asr of iNrliriwr awd 1)rertw 07 Hralttt Ser.- hre, Tulu Urw.et.ily. orAL Mw. PAULINE HEIZER. t'N.D, Rrw.d AuaYiotor Iw Cyr.~,~1 enI CrarArwdr (ry. Saw Frawcisce ~.rNwa of Medkd SdcsiceK San trawci.co. LAWRENCE L. HESTER, la. M.D, rro%aerr awJ Cq.irwwn. Deprtwwwr o/ ()lprriea and (:rwrsaloTy. Medieal CalleRe of SowK Caroliws. CKarkMea . G OSSMAN. M.D., ru.D.. A»orirrr (7hwira/ nofrsMM ol A/r/i- tir. University of Cdrferwia Medical (:ewler, Los Anoeles. Ettsl: CURTIS HOFF, 111 D. Ptr.D., f ra/rs,w owf CAobwaw. Dir/siew of /syehr.rk RrarreA, MeollIea1 CoMM of Virgrnra, RicMNiw/. CARI. C. ORU11711, MD. rsrD- Ar- sorlrrr i. Ihyswl.•tr awJ rhwnwnl- oRr. llniversily of rcnnsylvania (Irad- ra/e Sc6owl o/ Medruae, tbdadclpYu. R(1SSF1 1. 1.. 1/(M MAN. M D. Laolrl- ana Slate (/wrversMy School of Mrd1- crne, Ncar Orkans. 10)

I OLF. A. 11(N.TERMANN, M.D • Rr- AT7 Al I A/l KAPPAS. M D. rrolrssor rrarrA S. irnrur, 1 oArwd / aAwarwy, .wd Simw r6r..ri.w, The Rucfefelkr Umvcrury of Norre D.w.e, Norre Urvereitr, New Yorl. 1)arnt. Ind. FREDDY IIOMSURQFR, M D. hr./s drwt awJ D.rr.ro., RwKesearc` Iwsll- wee• Inc. Caw.bridge. Mass. ROBERT W. HULL, Pu D., rro/ruo+ o( swrlog.rol Sc.rwrr,. Florida S4w l)wi.ersiwf• T.Ralauee. HRATCH KASPARIAN. M D. Anut- owt Ihcn tw, Crdwr.>, alrr /itioro- torr: Iw.nrctor iw Mrd.rlw.. Halne- wsww Medical College and Hospul, Tbi1.JcIpW.. [UHU KATZ, Pp D., A»oriar rro/re- .or o/ S.xiulotr. University of CY- c.Ne. Clsicago. SHIRLEY 1.. KAUFFMAN. M D• rr.~ lr.w e/ raAolotr. St.re Uwiverdlr n/ New Yo.l Oowwu./e Med.c.l (.ew• k+• sroolly. ANCEL KEYS. PN.D., Dlrrrtor. I dor.- rrs o/ rbydo[~ Hygiene. Unirer- sirr of Miw.ew(s St1od of PuWie Heahk Miwne.pdis. IOSFPH R. KIRSNER, M D., rro/rs.o. of Mrd.rinr. Uwiversi/y of Clicato SJool of Medicine. Chicago. EDWARD 1.. KI.AIMFR• M D• Srwior SrNw/1lt, Tle WorceMer Foundation ror Eeperwnewial ttido{r, Inc.. Sbrcws- y~rrr. Mar. ~PETER H. KNAPP. M D•. ReMMci rro/tuor of hyrAi.trr, Souon Usi. .cru/r School ol Medicine. Ilouow. KENNETII P. KNUDTSON• M D.. Uni- .er.Nr of WavirNros Scdod of Mcdi- eine. Se.uk. ALVIN 1. KOSAK. M.D.• A..or(./e Pio%swr e/ Cirwri>Dry. New Yorl Uwi.ersiry. New Yal. ROSERT A. KUHN. M D, Auociut rro/rssor. Drrisiow of HrwotwTccr. New fersel Sr~ C~kie of Medicine. lenar Ci1f. STIO KULLANDF.R, M D.. rro/es.or ~ C~r"~. orprtmrwr of Okstra rK, and Grwrtulop. Univerear ol • • ~~ Swede.. 1'~ MARVIN Kl1SC1INFR• M D.• New Yorl Uwiversily Medical Cemer• New York. CHARLF.S W. LaRFLt.P.. Pw D, A.dn- oMnr leAtrso. Medical ( ollclc• Pbd.• ~1Plia. RONALD R. HUTCHINSON. M O. Rw u.rrA Dwrcror, Fwrndarww (or Mcis..• iar.l Reeeack Ao~.us Mid. /IT RFSFARCII INSTITUTE, Cfkye. ALPIMNSIi 1 . (N(:FN1TO, M D. Ar.r 1. cwtr rralrrwr o/ rArrww . do~ Eas1 ( aro/iwa Uwiversirr Sclod ol Medl•cfoe, (3ree..ilk• N C. HARRY L . IOACIIIM, MD. Anrwhwl ra.Aelo~nt, Lewa HIR Ho+pwal ( Iw• rd rrulirwr e/ r.rAslo~t. Co~rwwba Uwi.erw~ Cdk~c of hlrsKi.ws i S rrleow.. New Ywl. OEOROE /ACOSS[Nl. M D. ho/.uer and f/rol. Drp+r ....rwr o/ Rslw/opI• UwivasNr o/ Sowlerw C.tdawlaScbol of M.dlciwe. Los Aw~e/e.- JERRY HART JACOBSON. M D. W- nder. . Division of t IrctiopAl.blop• New Yorl Eye and E~r IwMw`arr. New Yorl . lUL1US Il. 1AC01l10N !1. M D. Awtrue rro/r.wr of Swfr.r a.d Dirrc.or ./ SrTir./ Ar.r.rcM. Uwi.er.iy d Vatwror Celfeie of Medici..,, IMr- ~a. MURRAY E IARVIK, PwD. Aarrl.u Pro %uor of rArwsao/op, Ateert Efr Mei. Cdk~ of Medieiwe of Yerli.s U.irenNr. 'x RrowR H. Y. OSwA1.D R . . Hk(ES. MD. St. Lat.Y lleqqaf, New Yal. W/I.UAM 1. /USKO, M D., AuetMrr rro/ruo. o/ liwrwr.rewire: D..cwr.Cliwttd rArw.aola.nk. L.ia.ro.r. MiR ar1ar1 FitMrote Hospital. R.rll.b. ANDREW A. KANOUTS( H, Pw D. Sa/ ScMwr4+. 11e l.clsos L.lor.. lar . R•r HNbor. Me. ARNOI D R. KAPLAN. Pw D. DDer• to.. l.ohor.ro.l of Mrdkal Grwrnr.. ('kvtland P.YcAuvic Inuilule and lloep•u1, ('levcland 101 I AARON I LADMAN, PuD, rrn/rsror awJ ('harrmew, Drpartmrwt ../ Awn/- sit"r. '1)ie llniversNy of New Mcsico Sl'/11M1 .// MedlClwe. AlbY'Verqye. THOMAS C. I.AIPPI.Y, M D, rro/ri- aor of %rtrolotr, Norll.weslerw Uwi• .ersilr Medical School. Clicqo. MI('11AF1 F.I.Al1M, M O., rro%,..~r a/ ratb,.ln[r. New Ywl uwiversiq Medi- eal Center, New Yorl. PAlll. S I ARS(1N, PN D.. Ho.R rrn/rs. sw of rhrca..w.d..er. Medical Colkie o/ Virsiwi., RicAmwa/. R(X)FR K. 1 ARiON, M D.• Chief of M.Iiriwr. Fu.wo Corwwy IlospMd, Freswo, Cal. OUS'/'AVE A. LAURFN7.1• M.D. Chief of Mediriwr, S(. Vincent Hospild, Worcester. Mwss. JOSEPH M. I AUWFRYNS, M.D. hr D • h../rcwc O.diwanw and (Y.MC- wr.R. Drpwrtwww. o/ rrhnl..Rr end Ml.ro>n.rk Aw.rowsr, EsPerbwewlsl Laboralo.y of /ldnrowsrr /IwopasAot- oar, K.Mn/icle UwiversileN N LeYrew• Le.vew, lklsiww. EDWARD I FP.TE, Pu.O., DSt., lro/te- .er ../ CfVrmifnr. Uwi.erwr of Miwne- sas• Minweapola. RICIIARO A. I.FRNP.R, M O• Acu.riote MrmAer, Scripe Cliwie awd ResearcM Fouwdaliow, 1.s lolls, Cal. CF.CILR LEUCHTENREROF.R, PtrD., Hrad, Drparrmrr o/ C'torirwrbnr. Swiss IwslNwe for F.aperiwreMal Csw- ee. Re.eucit, l.wsswwe, Swirmiswd. AVFR111. A. LIFSOW, M.D. CAdr- maw• Dernrrmewt of %rAo/.qr. Yale Uwiversiry Sclwd of Mediciwe, New Havew• Cown. FSTFN O I INDSFTN, M D.• PN D.. S(. 1.nepA's H...piui Rese.rcl LahorMott, S/. Psnl. Minw. ROSFRT 11 I.INNF/ 1.• Pa D, Aewt/- wr rra/tu.w of Clumi.nr. University o/ Vetw.owl, Rwlinpow. HERBERT 1.. I OMIIARD, M.D • M P If. . AI/ihrtr. ('an. rr Arnra.. A/w- u/rrr, New England /)eaconess Ilus- p1a1. Boston. 1. P. LONO• Pu D. rro/tn.r of rAr- wnwo.lorr. Sure University of 1ar. College of Mediciwe. lows Ci(r. CLAYTON O. 1.OOS1.1, Pt(.D. M D Ha.tiwr. rro/nw. o/ Mrdrriwr wJ rath..h.Kr. Uei.ersitr of Sowlser. Cdi- /o.wis Sclous of Mediciee, Los Awpdes DONALD R. LOl1R1A, M.D., Asaoclre ho%.soro/ ltlediniwr. CorweR Unlwr• skr Medical CoRqe, New Yoel. KFNNF.TH MERR/L1. LYNCH. M.D, St•.0., LI..O, Iro/tuor F.ww.4w e/ rwhoAr[r owd CAowrellor, Mediral CoReie of SaNi CarolMs. CMrkelew. INFS MANDI_ PH D., Aaluoru rro/r.. MK INM'iMNI)trr, Columbia UM.ee- Neri. Yo.l, of Physicians i Su.w ow. IOHN H. MANHOLD• /s., D M.D rrr.(r.wn owd Drrraae• DeP.rrwrrwr ej rwAat..tr owd Ord Di.rwo.is. New /eney College of Medicine swd Dew. risrry. laeey Cirr. DAVID E. MANN. PN.O, A,.x1w ha/easer o/ ri.rwrwco/opr, T.rwgil. Uwi.enNr School ef Plsrwser. Mi.. d.IplrM. FRANK ARTHUR MANNING. MD AlNUaor rr%aor of nlarn4l o;j Grwrt.rb~r. Wowrrn•, 11od Aw{eks (`w~M~/tlwiverswr o/ 5 wtte~s C.lifwsi. MNieal Center. Lae Aqdep. IOHN P. MANOS. M.D.. Iww.rtroe 1. 044 ~ ~N lofoRr. Medkd C~ >~ S oliw.. Cb.ekws. CHRISTOPNFR M. MARTIN, M O IrHUrM rro/r..nr o/ M.Mrlwr s;j DMrNer. Dirieirwn of /w/rrtlwo D/i, r..r., Se(aw HaR CoRqe ef Medielwr, leney cilr. R. RUSSI?l.l. MARTIN. M D., rro/nasr o/ Mrdk/w., owd Afkroilolorr, 1/w.. MArmalotr. Saylor CoReie of Medklwe. Houston. 11A5ON RESEARCH INSTTTtRQ Wurceelee. Mass. DONAI D 1. MASSARO. M D.. AcwA- arr rra/r.rw e/ A1rd:riwr, Oeor~. Waehiwrew UAiver.ilf Sclria/ ot lkdiciwe• Wns>.iaKrow. D. Cr ('/1AR/ FS MCARTIII/R, PN D, rcr- rA.d..Kiv. /Iwlrrrptl Ilrolii Serrka, //arvard (lniversily. C.rnMidie, Maa. 10S

.. 7D .l.. , i 1 t t : CNARIFS If McfANTS. PrrD. Arw- .rwrr r...fru.+ of S.wl.. NnrrA Cato- l.na Save Cdk[e School of Apicvl• lurc, Ralei[h HFNRY C. McOll.l. 1..• M D. ArNey llraJ. Drprmwnr of Puhoi.Kl. l.ow• pawa Srare U.i.er.iN, School of Medi- caee, New Orkaws. IIFNRY D McINTtKH, MD. hofrr NN of MIIK/M anI DI/r.IM, CrNs- r.wrlw [.Lo.aror,, Du•e Uwi.ee.M) Medical Cewter, Dwliam. N. C. FOROF. A McIVPR. M D, AcweWr rro/rasnr of rar/kdo[,. Medical Col- k[e of SowA Cato/a... Cirarktion. POWARD McKPP M D, M./rwr awd Acriw[ CAarmaw, Dr/rmwN o/ rrAolo[,. Medical CoRcqe ef Soah Card'ar. CAark+row. KELLY T. McKF.E, M D, Arsoclete Professor o~ M.Ik/wr. Medical Cd• kne of Souw Cardiw.. Clwkaow VICTOR A. McKUSiCK, M D.. rre/rr• aor of Alrtiriwr. The loMs H r.r U.i.ertity ScLool of Mediciwe. a1M1• wwre. /AMI:SO. h1111FR, MD. PaD, rro- leslur ../ r./y hi.vr, anJ Dr.r.hr, Alrnraf Ileolrh Kr.rar.h /w- aitrte. UniverW) of M.cA pn, Ann ArEor. CHARI.FS MITTMAN, M D. Dnecrnr. Drprl.nrM of RnrLa.wo, Diuares, Cirr of Ik+w NNional Melecal Ce.- /n, Duarre. Cd. HUON MONTGOMFRY, M D. Ano- efur rr.•/iu..r of Mel.riwr. I/eiversNy e/ Pewwrplvauia ScMwl of Medicine. P1i1.JrlPlwa. P. O'l1. MONTOOMFRY, /.. M D, Professor e/ rarA.dn[j linivet.il• of Teer SowAweMerw Medical School. Ddb.. OEOROE F. MOORE. M D.• Pu D. Di- recror. RusweR Pwt Mewwrial /wtl1- 1we, Rullab. N. Y. KF.NNFTN M. MOSER. M D.. At.i.awf rrofr.rnr ..l MrJirN.r. (3ra[eloww U.i.ersirr Medical School. Washi.!- le,n, o. C. HURI FY 1 F.F. MOTI.FY. M b. rroles- Y.r, of AfrJM11K and DMrrIM. Crd1o- Rr.puatws fwhorro.l. Urrversirr of Sowhern C.Idornia School of Medi- ciwe. I otr Myeks. EDMOND ANTHONY MURPHY. M D. Sc D.. A»ociatr Professor o( !Na/Nirsirs and Mrdicfwr, The lobM HoNiw+ U.iveniry School of Medi- cine. mahirmore. W11.11AM S. MURRAY. Sc.D. Senior Sta/ Scirntirl. The laclaun Labot.- 1orf, Bar Hareor. Me. RICHARD L. NAF.YE!. M D. Professor .wd C/yirm... Deprr.nrnr of rarh.d- ep, Pe.ws,i.aria SrMe Uni.ersNr Cd- kspe of Medicine. Hcrsbe,. OF.ORO 11. NEURAT/I. PM D, MNro- awd,rk./ [.doraror,, Ilameut[, WeM Ocnnawr. AI SFRT 11 NiDFN. M 0. rrofeswr.l 1frJwinr. Dtew PuM4raduNe Medicd ScMuol and UwivcrsNt of Soutbeew C.lif.w.ia: Cbirf, rwl.nowartr Dbrau Secriow. Marliw I uwbcr Kir.e Ho.plal• I oc Angeles. OAK RIDGE NATIONAI. 1.AOORA- 7()RY, Oat RidIe, lerm. ROSS L. Mc1.EAN. M D, AnorWr Professor of Mrdicinr• Fmwf Uwi.et- .A1 School of Mcdicirte. Atlarwa. OERALD F.. MdLFARN. PMD.. Di- rrclor. MqbMr lo. lrharinrd Gr- nrrkr. Dr~.r/wwwr of rsychdo[t. V.i- .ereilf d Cdotado ScAoo/ .1 Mu- wracf. moulder. WILLIAM F. McNARY, 1.. Pw.D. Aa- aocirr Professor of Ammotor•7. 90~ UreenirT School of Medicine, Ila(on. NFAI. L. McN1VEN. PN.D. The War- eesler Fowd.lioa for FaPecnweN.l RioloRr. Sircw.Awy. MasL IULIA MEYER, hr D., Asrorirrr rr.- /..rer of ()ra/ rarAofo[r. U.i.eesity of 1Riwois Co1kLe of DewustrF. Chicago. pOV MICNAEI.1, Pw.D.. Ardstunr rro• /rruw of lbrhe.wlsrr, .wd SMNr,. uaversirT of (-aliforrda School of Medici.e. Sa. Ftawci+c.r. RP.RNARD / MII I FR, M D. ArdrtoM Professor of Aeuro.n,. leflerwe Medi- cd Cul/ege, PAiLdcIPAie. 106 i UONAI D M PACh, Pw.U., rro/nsor '/ rhlsadu[7 onJ Dararor, lnuo.re l~N Cellrl.r RruortA, U.iver.Ny oP Ncbra%ia, l.iecub. K"NNIiTII RAIGI:N, h/1)., [laevr..r, lMiwrnnrnr of M..k. r/.r RN.4.[f. Iicarrh ResearcA, Iwc~ Ru.weR P.rl Divniua, YuRdo. AI-sERT !. PALMER, rw.0.• Auiuawl Professor of rrA.Jo[,, U.iver.iq o( TukJq 141111140. O. ROSN MAR/F. PAN(7110RN• MS., As- .tuowt t"o..f TrrAnolu[i.t .wl Z !.n hrrr, r.rrMrM of f•'ouJ SlNntr ow1 Tr.h- u[,• Uwirer.iry e/ Califoc.ia, D.vie. /O/IN W. PARKER. M.D., Aeeeclre r..r~ra.e. cr/ rrAulo[). U.i.er.il, of SuwArrw Califorwie Setiod o( Mea a.c. Loe Atycka. MARY STEARNS PARSHLEY. PN.D, AsNssont rrofr.aor o/ Awroiwr~k arn.k. aw/ Gywrcolo[) C. Uwi.er.iq Cdk~a of PMy.iciawe ! S.r. geowmNew Yort. EDWARD W. PELIKAN, M.O., CA.M- wrn. Drlr*rww ol rArrwarel.[, owf Eyrrawrrua/ TArryrwk; NoMew Ua.p.i(F Sc1ad ./ McNici.e, Ro./o.. MALCOLM C. MKk Pw.D, rre/naa. o/ CowrwrrwMr Mrdici+.r awJ rrJi4- rirs. Universi/y of SoMberrra cali(.ewi. Sebod o/ Medici.e, Lue Aqelea. OTAKAR 1. lO11AK, M.D. P«.D, Eaerrthr Di.rcar. Dmm Metrical Re- eearcd Cewuc, l.e. Do.e[, Del. MORRIS POLLARD, Pu D.• DMrerer Lubrwl [.br.ror~~ U.i.erei(T .~ Noire Drwc• Noue Dat.e, IN. C. M. POMERAT. PM.D.. Dir.ttnr of liolopkof Rrs.rcA• Paudewa Ferr.d.. riow for Medical Rewr$ Ps.dewK Cd. S. N. PRADHAN, M.D. Pw D.. rro)ra- .ur of rhrmw ofo[~, 1lowarJ Uwi.er- wf CoReRe of Me.Mciwe. Wa.IJrytow, D.C. H. R. PRATT-THOMAS. M.D.. rro/r.- aer of rarAolot, aeil Draw, Medical Co11eAe of Sow, Cardrwa. CAark+row. PROCESS 11 INSTRI)MF.NT CORPO- RAIION, Ilroollyw, N. Y. MARTIN S. PROTZEL, D.O S. CAk/, Drplmrwf u/ Urd ruAelo[l, Newart Ciq Horrild, Newarl, N. l. WAI.TI:R REDISCH, M.D. A.udne rrOfrJM/I of C/iAi(.f Mr/ki/N• Naw Yort Uwi.ecai(y Scltooi e( Meliti.R awd NYU Reserch Servioe, Odd.a(p MeworW Hoyi(.L New Yect. TIMOT/IY 1. RFOAN, M.D., Professor ./ A[r.GrMr Dirraver, Dlrisinw of rr4hor.Ka/./ Disrases, Colle~ a( Medicine awd Deai+uy of Ncw /ereet, New ler.ey Medical Saiool, Newrl. WII LIAM RF.OEIS(M, Mb, rn.prw a../ CAairsw., Dr'.rrnwwre/ Mr/ki./ Un.ol.q,. Medical Colkle of Vir0wl., RicMawd. LYNNE M. REID. M.D, N'dl.rA fti.. rsw.r u rarAuto[_r, /l.rvr~ Medlcd CA.rrwran, D.rnrwewr ./ rrhulo[,• ChiWte.•e HoqMr Melital Cerrer, Bea.., HOBART A. RF.IMANN, M.D. r., aow of AIrI:cMr, H.Mewr... CeRep .wd Ileyi(d, Pbiladeybi', R01.LAND C. REYNOLDS, M.D. A.r waM Professor of r.rilelo[,. Udeee• .il~ af Teaes Sowf.eMaes Me/ied ScAool• D.Ra.. VICTOR RICHARDS, M.D. Chief of srsrr,. Pretky(e.iM Medial Ce.(ar, S.w Frawcirc.. ARNIS RICHT8R5, lw.D. Aa.aire rn/rsw of ratI1r+fvaslq a/ So.IRer. Calilaw ~oO1.1 M.dIeMK L.e. Aryelq. WILLIS N. RIESEN, PN.D Srw4r No- cAr.r/w• Life SNr.cri &L/rMq IIT Reaeard Lwilwe, CR{cy., DANIEL R. RIFKIN, PM.D, Arirr rrw/ruuw o/ CArndtd !1.1ee RodefeRp Ua.erWr, New V«!~ R. H. RIODON, M D., rrojra..r of r.. tA.l+[f. U.heraM, .f Te.w Mdltal rr.wcl, Od.eNaw. SYDNEY C. RITTENRlRO hD • rru/ruor o/ uf Sotwherw CdN~or.ia~„ Le.P~ . Ayd... RENS(Ki • ROE. M D. A.rrl.re rr. Irl- N swprr: CAir Crdi.r Sr- [r.'• Uai.ecwf of Ca~sfor.la Stled of Medicine, Saw Fcan:isco. 107

/ . . 1OSFP11 II ROOFRS- M D. Holy N.me of Icsus Ilospeal- (7ad+.kn. Ala. ROSF.RT C. ROSAN, MD.• Aworlott rr..fruer of rotAol.•tty ewI rrli.rrirs. S.. t ouis Uwiversilr School of Medi- eine; Arsoci.rt hriessloTnt. Cwdina) Ok.now Mew.ocisl IlosFi.el for CYT- drew, S/. Loris. CHARI.ES L. ROSN, hs.D_ Cl/nlr Dh+et~ ror; Dirrrrr, Norw..Nre Aa4.d Srnfy. Veterans Adswinislrslion OwFs/kd Clink• floalow. IMIN A. ROSF('RANS- M D.• Au.- (.re rr.•fruor ../ rA.r.wer.d..ar• Mcdi- ca/ Cdlcge of Virttiwla• VkRiwu Cows- w.ewwealU' Univerw7• RklMnowd. IOHN R. ROWI AND3, hID. S.el SrMwricr, Sor/bwesl ResearcA Iwalkuls. Saw Anfon3o, Te §FNIAMIN A RtlSIN• hM D., Ass4t.nt rrofr.o..r of raANr NralrA, ea>lo. ttwiversnf Colleae of Medaiwe- Hou- saow RONAI D P Rt/StN. rw D. r.ofruwr o/ rhr.wocolotr. McdKal ('dkgc of VMlInia, RKIllnond HFNRY I RIIS%I K. M 0. hruJrnr, T1e Russtk Foundation. Inc . Sfstew 1s1.nd, N V W. C. RIKSFt 1, M D. University of Te.s Medical (-cnar- Il.wslon WAYNE L. RYAN. MD. rr-legwr of IliacArwriur7. UniverafT of Ne6casla CoReRe of Medaine- Omab. t•FTFR F. SAI ISat/RY. M D.. tM D_ Nrrl. Inftwsirr Treorwrrwr Crwrrr. Sais( /our. Ha,pi(al, eseb.nl. Cr. •Attl. 3AI.TMAN• hs D.. Asslsu.nt rro- tasnr a/ /JorArwe/snT owI Nwnlriow. wi.ersit of SowRerw Cdifornia School olMtdicise, I.os AnFeks. I/I RICH H. SCIIAF.FM. M D. Dkrc- we of Nrwa.~..rwr.reWfp. Maww Re- aarc IwsliNwe- Worfasltr, Mass. IOR(i114 U. SCt1I FOFL• M D. h.D., rrefrss.r ew/ ('A.irwww. Drp+rrnwwr of Sretrrr. Tulane University School 01 Med.ciwe. New (hkaws. AI VIN N S( /1M11) t MI). ll..r.1.w n( f.•..w.rl.~t lofn ltw..r.aa. Mtd (.NJ. t.(ass IAKOR SCIIMIDT, M D. PRt D., As- ' si.ranr rr.•(r.a•r a/ N1.w llr.ni.nf. Di- r/lMM of Ib.d..Ku.l .t.ir•wrr. Stale University of New YorL at Slonr RrooL. S/onr erook. ISAAC SCHOUR. D D S- hf D, D Sc. Dean. Uwivcr.ity of Illinois ('o1kRe of DesMisuy. CYic.{o. SCRIPPS CIINIC AND KFSFARCII FOUNDAI ION. t. lolls. Cal. MAURICE S. SFGAL. M D. Cliwkol rrelrrMw o/ A/rJMlwr, luUs t/nivet• .Ar School of Mediciwe; nMecr.», Dt- rrrw.rws o/ lwAwI.~i.w TAruIy, Ilos- w. Cify Hayw.l, b.wow. CARL C St1 T7F.R. h. f),. N.•worrr Rrsee..A Arw...rr. Peabody R/useuws, Henrd Uwiversi/f• Caw.Erid{e. Maw. LUCIO SEVERI. M D.• Dirrcrr .wJ Draw, Iwuirrre of Awernw.l.nd rerhol- z ,• Divisiow of Cancer ReseascQ arvetsily of !'+eruRia. reruRia, Ifdy. CHARLES R. SHAW M.O. Chirf- Src- r/on of A(rJird Grnrrirs- M. D An- dersow Ho+Fitd swd Trnan Inssi.ule; rr../r.wr 1.1 Iholoay. Ile University of lesss al Ituuuow, Ilous/on. C//ARI FS F SIIFRWO(1D. M D• Ar .nuwr rr..fruor of R.Ja.f..ar. Ualivet• ufr ol K..Acdcr ScMrrl ol Medicine and tknm..uy. Rock+lrr- N. Y. SIII)1/ S11tKATA• M D., Pu D. rrelrs- wr of r~~doa1. University of IlawaN School of Medicine. tlonolulu. DAVID L. SIMON. M.D.. Iwurartw 1ot lwrrrwe/ M.Ikiwr, Cincinnati Gtnerd Hos'i(al, Ciwcifwnti. ERIK SKINH0/, M.D_ Chief. Deprt- wwwr of Nerrolop. DiveAjets Ilospi- lal. Copenhyen. NATHAN H. SLOANF.. hs D. rrn/rs- sor of /iorkrwrUo7. TAe t/niveosisy of Tenwesses ('ewler for Ire Heah` Sciewces. Memphis. TIIFOD(1RE A. SI.OTKIN. t•a D.. As- si.Iewe rro/tssw of rA.r.wocolotr)•. Dule t/siveesitf Med.cal Cenur, Dw• b.ns, N C. (;1'()K(T w SMFTtFRS• M D• Aua .urr .w rr.h..l..~r.No.l~wcslerw Unl- .cr.a r Med«cal Sc~..ol. ('raqo. 109 S t GENE M. SMITH. hr.D Aarirronr rro- /rswr of rsyrholotl. H_enrud Medical Scbod. Masseclusq(. Oe.eral HosFi. /sI. Iknlow. LUCILE SMITH, ho.D_ rroJrssor o/ .iecAtrnisrr,, Drt.ow. Medical Scbol, Haswver, N. H. SHF.LDON C. SOMMERS. M.D_ Dbn- rr of L.rrorerwkr, Lenoa HiR /los- Iw(ai; Cliwke/ rrojrsser of rNAolo=1. Columbia University CoRqe ef rbysi- ciaws & SurReo.ti New Yortt. ERNEST SOND/IF.IMF.R, tM.D_ Asis. c/.rr rro/rssor e/ RiorArw.Jwl. Col- kse of Fo.errl. Stse University of New Yo.l• Syracras. T. M. SONNERORN, he.D, Diniw- rdsAr/ Sarvk. I essr of Zeotoj*7, Iwdisws Universi/f: oo, SAM SOROF, h/.D., Ne.1. Dt'.rwuw( of Arrarroswe/rcrlr CArws4/r,. TM Intwi(ws for Ca.cer Ratuai, 1'fila delpleis. SOUTHWEST RESEARCH INSTI- TUT1LSa. Aro.ier Tea. DAVID M. SPAIN. M.D, Dintrr, Dt. r+.rwwwr o/ rrAdop. ?1s komldd. Hoyi(al Cewus, Rtooilrq, N. Y. ALEXANDER SPOCK. MD., Aadawwr rro/rur of rtdi.rrk; D.Ae UOirse- silr Medical Cewla. Dsrkr, N. C. FREDERICK 1. STARE, M.D Ire/ts- aw of NrrrM1vw, H.rsed Joi.uaAr Se1ao1 of Public Iled(h. ReMo.. C. HAROLD STEFFEE, M.D. Drrtler M~.rorks, Me1RodYt Hoqi1.~ JACK P. STRONG. M.D. AsaselMe /ro%ssw o/ %rAofftf. Lwidw SeNs University Schod of Mediei.e, New Orkews. MARION R. SUL2BEROER, M.D, ho- /rseor "I CA.irwwn, Drp.rrnwrwr ./ Dnw..roforl and Slpliilu<ely.. New Ya\ U.ieersiqdlefkwe Medical Cera• /et, New Yort. RENATO TAOIURI, PM.D., Aaaft4Ye rro%ssa+ of rsycAoloa7. OrduMs Sebol o/ Rrdwess AdnsiwissrNiow, Ila.ard Uwi.enwr. /orow. DAVID W. TALMAGE. M.D. Dbecrr, WrN-WriwT Lasd Iwskwr• University of Cdorado Medical Cenlcr, Denvcr. MARC D. THAMES, M.D. Sedor Rs- srrcA Fe/low, Msyo Cliwit ad F.r.& drio., RotAcsar, Mir. CAROLINE BEDELL THOMAT. M.D, rro%ssur E.werlrrs of If.Iklwe. 71s loMa lloFt _ifa Ua~fvawf Sdwd ef Medidwt. ~aNi.ors. JEROME F. THOMAS, Pr rr D tose of _.S.oAurl twRiw.erlwR. Uavtn ef (,aNfetwiq Re.LeMy. IJAMES E. P. TOMAN. PaD, tAye.. aor alwd CD./rwsow. De/urtw.rwt o/ rLr' w..colorr. CRicaRo Medicd Scfool, 1. MNutr (er Medical ReaeatcR (ycy.- IANET TRAVELL, M D. As.x1w rrrr/far. .~ Clhk.f rArnseco/.q. CoeweR Uwistdl7 Medicd C.ReK New Yort. LIE SHA TSAI. 1•w.D_ ReseaeR Aaf+ cl.r. M r.r/lofodl. Yak U.i.asMi! School of Mediciwe, New Il.va. C.r. OERALD M. TURINO• M.D., hoJesrr of Mdreiwe, Cf11u.Ws University C.F Ylap efr!•~ Frrsieisra A S.r....q Nsw D. M. TURNER. M.D., Beu/, D.rwr. /wrax o/ Dr.r dfaot•eqaiw sud Nw rAews4trY, Hatlelow Lasaewti.. E.- 1~ LW., Hrro:.ls. YstLsWs, Ery- UNIVERSITY OF SAN FRANCtSCO, Satt Frsaci.co. UNIVERSITY OF SOUTHERN CAL!- tORN1A, Las Asyelee. EU.IOTS. VESELL, M.D,"ep..../ CA.f.w..w. Dq.rrwerwr eJ Iherfw.rd• on. rew.srl.ew S(sa UMivaeilr C.1- k~t s1 Maiici.e, MiMfire S. HsnMy Mc~ical Ce.(er. Ncssie7. RRAN/SLAV VIDIC, D.S., ho%...r of Awaow.T. OsotReloww UnieatsMr Schoole of Medicine s.I Deolrry. W.UffRtoq D.C. ROMEO A. VIDONF_ M.D, AsssKl.w rretaaw e/ r.rAof~ r. Yds Uwl..r- si(. School rrf Me 'dsc~, New Hs.ef• Cores. PETER K. VOOT, hl D_ Irofru.r o/ A(krobqMay. University of Waslii.t (ow School of Medicine. Se.nk. 109

j E 6 WARNF.R. M D., ho(r,wr o/ re• tholn~~. Su1e Uarversi~f of Iowa Col- kfe ol Medicine. Iowa Crr. SH1F1-DS WARREN. M D., DMrrtr ol ['sbw."w.rr. C.p. rr Rrrrrrh lw.n- t.a. New E.41e.d Deaconeu Hoqi- ra1. 11ouo.. YASUSHI WATANARE, M D. Asr n.rr MrrwArr. 7w WiNw InaMw ef Awuawr sad sido4l. rfsldclpisia. BARBARA K. WATSON. hM.D, AaiM- .wr /.rrrr/elors/. Mauaelso.elr. Oen- eral 11o.iilal; Rr.rrcA AuerWe i. /.rwrbluor .nI M.w..wolfty. Hr• •ara MeAkal Sclsoelk a oeron. LEE W. wATTF1leERO, M.D. ho1r.- ner ./ hrheloty. U.iverwT of Min- .esele MeAical SchooL Mire.Foli.. JOHN S. WAUGH, M O. ho/rw o( CArndrrF. MauacMnaru lar Mirwe Ted.elop. C..beidr. RICHARD L. WECHSI.ER. M D., C'R+ k.f rA7rlo/orla, MowreAore Hwvitd Inuilwe of Re.eucR tin.UwN- IOHN V. WEIL, M D. Artlu.wr rro, /r...r of M.Ikiwr. Ua.er.Ar o/ Ce1. eals Medical Ce.ter, De.ver. A. WEINSTOCK. M.O. Rr.wcA /1.- eAeawn. Ule Scwwcti Di.i>t.n, IIT Reeearcd /.rirwe, Cbkap. RUSSELL W. WELLER, 140. F."- .Ou, MeaworW Ho"sl of CuMer CooMy. WeM C%eree. /a. A. STANLdY WELTMAN, fM D.. An. clMe he%aer of rArwucole1l.ad Rr...rcA. ModIP• CoRep o[ M.r- .waey. Rro.UM N. Y. SIMON N. WtNDFR, rn D. RnrrrA rr../r..w of aw.hr.nunl. Univeraity u1 UlieAoma, Nonnan. DUANE O. WENIEL, M D, Iro/rsor rel Ch.irnuen. Drpwtwwwt DI Ihrrns.- tolo~~ awI To.icolar1. TYe~Lniveni~y d Kaaus School o/ Mum.cr. Law- rewn. THOMAS C. WF.STFALL. htD.. Ir..- Ieuar ol FArwarul.rrr. Unsver.ilf of V YRlda School of Medrciwe, Cyar- knes.i/k. FREDERICK E. W/IISKIN. M O. r.M., oheetr. DirJ.Mw .I Nr.ith ..J rrr. rrweNry EprqYelrrw. T1e Ar CeMer el New E.Alsnk l.e, Rowa+. ROGER J. rY1LiJAMS, M.D, rro/tuor of CArwdury: Direa.. Clqtow Forw- Iaie. /.orArnwcd IwwM.w. Tle U.i- venily o( Teaar, Ar.rin. ;q DANIEL H. WISEMAN. M D.. Auix- ..r rro/n.orr/ reli.t.kr. UnivereiV e( Sowber. CJifor.i.• Clsi:bei e D'a sri.ioR Los Aiyeke Cowry Oe.erd Hoyilal, Los Aaecks. 1. EDWIN WOOD. M.D. Iwtnurtor iw Mefkiwe. b+eow Ural.er.Ay SAool e( MeAici.e, toeros. SUMNER WOOD. la. M O., At.la.nt rr%uor o/ LrAo~7. Yhe Ie+MM HoNiws Uni.enhf Sclsoal of Medl- ci.e. Rdli.sore. JOHN E. WYATT. M.O.. A1 Jnir of rrAdoEl. SsiM LaJ. wiver.il7 Sefsel .t Me`eYax, S1. Low.. KOII YOSHINAOA. hrD, Center /or EoNlalie. ReaarcR NICHO, NMk+.d LrMwes ol HesNA, Snbnda, Md. 110 >> I INDEX OF PRINCIPAL INVESTIGATORS Ace1o, M. D., 49. 50 Arco.,1. C., 12. IS. 19, 20, 21, 22 Arnat, M. S.. 7.8.16.24 IBaer, 1.., 50 'Senedicl, W. F.. 16 Sing, R. 1., 41. 42, 43 Bossf, R., BI, 82 Snshce. D. 1... 2) Csmrell, E. T.. 14 Carlro, A.. 69 t'hao. F. C.. 41 Cochrane, C. (1., 70.71 Crn/s. r. T.. lr.. 82. 1) ('nnnpaeNer. D. W.. 83.84. le3 [lowney, 1/. F., 44 Foirer,l. A.. 57. St FrieAman. G. D., !6 Fu.e, K.. 31 Gcotss, M. C.. 31, 52. 39; 71 (Teueer. T., 1) Gickn. l. E., 9.10.15.17. 11 Gleich. O. 1., 72 Gorrod, I. W., 60.61 C:w1oo, 11. L. 1) 11aR. C. B.. 29, 30 Hatrosh, r., 30 lleiawra. N. W.. 37 liou, W., 53 1aReoi/o, A.1., 62 IseoQ, A.. 31. 32. 33 luhmon. D.. ) l lu.lo. W. 1., 63 Kkineneas, 1., 34 Kdwi,.E., lawrencRe. E. C11. .34 Le.y,1. A., 72 Lynch, H. T.. 27 Maniw, R. R.. t, 16 McC lea.n, G. E., 54. 35. 56 Meier. H.. 26 Michseli, D., 64 MilcheM,1. A.. 64 Mwad, F.. 63 Or+ch, F., 24. 25 Rsela.ab, 1., 87.88.89 Repw, T. 1., 45 Reywolds, H. Y.. )3, M Rysw, U. S.. 36. 46. 47, 48, 73 Ss.rry, R. V. R., 66, 67 Shtlar, O., 26 Sare/, T. E., )7, 38 Towrelsko, A. M., 68 Trs.i..1.. 40 Unuwe, E. R. 74. 75: 76. 77, 71, 79 vss den Berg. 11. 1.. t0 Weinb.we, G.. 39 Will, l. A.. 79 WoN, O., 21 YosGioup. K.. 56 III