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ANDREWS t.. .1CE PRODUCTS CAPITOL HEIGHTS, MD (K)

EXTRA COPY REPORT - Of THE COUNCIL FOR TOBACCO RESEARCH-U.S.A., Inc. 1982

Organization and Policy T>te Careil for Tobacco Ra.rrA-US.A. lec. Is 1be spoe.oriag .peey of a progrne ol research isto Suewbr of lobsooo un ..d health. It 1. tlt" out- powtt7 ol r orpnl:ation fofrd ..r1r i. i!S• by Ios.ooo s.suf.ctursn, pow.n rd w.rdawmes. Reaatsl wpoat bss bem An.ldy t6tougY a pro. Rum ol p.N.i.•.1d wrpplaw..td by ooWstb for wssaci+ wit• ieuiwt{o.e .ed LborswrM.. TS. C.arcll Io..a q.aw s.y n,arcU fscility. TU Sckatibc Ad.isory Soad ts Z1s Cor.ci www rtgul.rly 1o c.duN. .pplk.tio.s for research sappoct, jidgift tfsn..olely es 1lt@ e.un of scisstiie nrog1t .ed nMr..o.. T1. C.oarcU .+.rds ns..rch pr.w b i.depadea seiestists who .r. .r wnd oo.pkN wAestiffic fr.edo~w is oosducU.g tleir studies. Ur.nteea .b.. .n reaporiWe for repaU.g or puE1Wi.g t6e'r iodisp i. tb..coepted aie.- UAe m.oset - tlrough mediul sed .cie.tibc jour..b .ed socklhs. f F I 1982 REPORT o/ THE COUWCII. FOR TOBACCO RFSEAR®-U.S.A., t.e. WnLuw D. Hosss . ~ Cb.irsn. ~ ..7. . ~ .f... ~..+' ~abf m cn ~ '111E COUN('11. FOR TOBACCO RFSFARCII-U.S.A., iac. 110 t:..i 591h Street, New York, N.Y. 10022 m . m ~ J W ~ r r

S/aEN"1'IF'll: AI)VISI)RY IN)ARI) to The Council for Tobacco Research-U.S.A., Inc. a. u( Ikcember 31, 1982 I.EON O. JACOBSON. M I1., Chairmat losryh Rrjrnstrin Pro f,•stur of Biolosical Stient rs ( rmrritus ) I'rojrssrw tr/ thr 1)r/NOtment of Medicine (enrcritus) Univetsity of Chicago C'hica8o, Illinois RICIIARI) ). BING, M [). llirre un u/ ErttrrLnrvqul ('ardiukrtly and St irnti/ic I)eveltrpmrnt Ilumrn8ton Medical Research Institute, Pasadena, California I'tn/ritt.r o/ INrdit inC (rmrritut) l)nmvenily of Southern California School of Medicine I.cn AnBeles, California ROSWI7.l. K. BOl1TW[:LL, Pn.[). l'rtr/rta» ot OncokoRv McArdle Laboratory for Cancer Research University of Wisconsin Madison, Wisconsin DRI/MMOND 11. BOWDEN, M.D. I'roJessor and flrad Department of Pathology University of Manitoba Ilealth Sciences Center Winnipeg. Canada MICIIAEL 1. BRENNAN, M.D. President and Medical Director Michi¢an Cancer Foundation Detroit. Michigan 1OSEPH D. FELDMAN, M.D. Mrm6er, Research Institute of Scripps Clinic Scripps Clinic and Research Foundation La )olla, California WILLIAM l[. OARDNER, Pu.D. E. K. Nt.nt Pro/essra of Anatomy (emeritus) Yak l lniversily School of Medicine New Ilaven, ('onnecticul I PETER M. IIOWLEY, M.D. Laboratory of Pathology National Cancer Institute Bethesda, Maryland IfENRY T. LYNCH. M.D. Professor and Chairman Ikpartment of Preventive Medicine and Public Ikalth Creighton University School of Medicine Omaha, Nebraska G. BARRY PIIiRCE:, M.l). Amrriuan Concrr So.lrty Cintrnnial Research Pro/estor LMivcnily tr1 Colorado Ileatth Sciences Center (knver, ('olorado GORDON II. SATO, PH.D. Pro/essor of Biolt>ty University of Calilornia, San Diego La lolla, California SIIELDON C. SOMMERS Scientific Dira•tor, The Council for Tobacco Research-U.S.A., Inc. Clinical Pro/essor of Parbolory College of Physicians & Surgeons of Columbia University New York. New York Sekatlie Srafi.( The C.w..eil SHELDON C. SOMMERS, M.D. Sc•ienti/ic• Director ROBERT C. IiOCKETT, Pu.D. Research Director DONA1.1) 11. FORD, Ptt.D. VINCENT F. I.ISANTI, D.M.D. Associate Research Dirrtvor ~ Associate Rrsiarch Director DAVID STONE. Pu.D. Associate Research t)irector

Abstracts of Reports Fol{owint are abstracts, approved by the wthors. of reports on new resean:h ackmwvled6ins support from The Council that have appeared in scientific journds since publication of the 1951 Report. The name of the grant recipient is in italics. The abstracts are grouped under these headings: 1. Cancer-Relaled Studies. 11. The Respiratory System. 111. Heart and Circulation. IV. Neuropharmacology and Physiology. V. Pharmac.ology and Biochemistry. VI. Immunology and Adsptive Mechanisms. VII. Epidemiology. 1. Cancer-Related Studies MECHANISM OF ACfION OF BEN?d)/a/PYRENE AND NICOTINE ON HORMONE PRODUCIlON BY RAT PITUITARY TUMOR CELLS Although hormones have been associated with ieduction and ptopessan of m- Foms in many e.perimental systenu,lhe role of hoaeones in the process of initi.tiw and pnog reuiat of carcinoge.esis is.w ckaly de8ned as yet. lw the pesesw ancnrpl w .ndersund the mechanism of .ctiom of benm(a)Pyreae (BaP). a cyclic aomsfie hydevcarbon. and that of sticaine. the lobaceo.lkabid, the effects of these .gents on ppvlacqin (PRL) and growth honnone (GN) synthesis by rat pituilary tunwr oeMs in cuNure (GN cell!-) were studied. Treatment of GN cells with.icaline (0.1 •300 pg/ap neither affected the growth aor significantly s/1etel the general p.nerw of hmmone podaction in these ceds. BaP Y coecentntlions Rrerer than SpRhnl imeversibly inhibited the growth of these cells. The subkth.l eoncentntions o/ B.P. which did aot affect either (1) cell growth. or (2) amino acid tnnspoal or (l) rotal protein synthesis or desradaion, did however inhibil specifically harmone synthesis by these cells. More interestingly. concentntiota of sicoti.e. wbich did no1 affect either cell po.nh or hoxmonc synthesis. modulaed both of,these eeRuhY processes is drc presence of BaP. A concentration dependent stimulation of microsomal BaP rronoo.ylienase activity was observed in nicotine or BaP treated cells. Tbe effects of :fiese substances on aimulalion of BaP monoosylleaase activity seems 1o be additive. Nicotine also en- Mnced the associMioa ot adioaclivily (pRsttmsbly 1711B.P metabolites) with DNA i. /'111B.P treated cells. (t is concluded pist nicotine by itself did na demonwae any cywaosic e/fect not in/M.ence bunnone synthesis in G/I cells. However. ftieoti.e stimulated B.P inoonosygenase activily and the inleranion of /M1/B.P metabd'Nn witA ceHular tN1 A and also modulaled BaP induced inhibition o/ hormone synthesis is (;11 cells. Chakrabarli, S.. Nancs, S. D. .nd Aiiwns. D. K. SiocArnrira! md SfopAysiea/ Reseomh C-rirawns 101112):596-607, 1982. PFrom the I.baaory of Pfiarmacoloty. Harvard School of Dental Medicine and De• p.rlment of Ph.rmaolo6y. Ilarvan) Medical School. Boshin. 7

EFFECTS OF SELENIUM AND SULFUR ON METABOUSM OF BFJdZA>foIPYRENE BY HUMAN PULMONARY ALVEOLAR MAt.'ROPIIAGES The interactinn between trace metals .nd polycyclic acronutic hydrocaibons (PAlls) has attrneted increased irresesl recently since some Ir.ce metah, inchding seknium, have been shown to decrease chemical carcirror nesis. Due to the high abaorption e(ficiency of trace rnetals iw the hsng alveo)us.nd Me ability, of (wlmoaary alveolar maaopha6es (PAMs) to rn.iaboline PAHs, attenlion is certtering tww on the role of PAMs as a pnmary defense aRaiant teaobiotics. Foc the studies repxtod here, focus waa on the effects of aekwiww (r Na SeOJ and sulfur (as Na,SOJ on cytotoaic- ity a.d coejugation of bento(s)pyrex (B.P) by Aiwar PAMs. Rta.ks showed 11W, .Nfwrsh se k.iwn was fotind to be wwre cyroloak thr wlfrr, lhe cytotoaicity of both selenium and wlfur was greater for PAMa obtained by bronchopulmon.ry, lavage from wowsnwken than (nr thoae obtained lraw cipnttle amoken. At IpM sekniwn, ghr- tMMone conjr`atwrn of BaP was ewh.wced M bod wwoker and nowsmokcr PAMs Even though aeknwm roaicity cr be achieved itt PAMs at high levels (100µN'), it seems d+r this element may afford protection by i.dretion of detoaification enzymes. In- decd, die lack of sekniarn cytotoaicity, is swwker PAMs suggests a protective effect fram crganrne smote coeqo.ents, ind.dift the tr.ce metals studied hen:. Marsh.ll, M. V., Mcltwrxe, T. L., Srsb.r. O. L., Marrin, R. R., and Grif6n, A. C. In. Pofynrrfe.r A.owwic Hydoc.rbows F~G1 /nsen.arlowof Syw~otirrw cn Cherwital Andysis awI sioloticd Frt, Columbus. Ohio: Banelk Press. 1981. pp. 221 •230. OtAer s.qp.rt: The Robert A. Wekh Fardtlion and die American Can:er Society. From the Univcrsrry of Te aas System Cancer Center aW Baylor Cdkge of Medicine. I/ottston; Nonh Tes.. State Unirenity, Dento.. ONTt>CFNET1C VARIATION IN RAT UVER. LUNG AND KIDNEY Mt1r1(IOXYGENASE INDl1CT10N BY LOW DOSES OF BENZO(a)PYRENE AND CIGARETIE-SMOKE CONDENSATE 1n previorn in vlnn qtrdits. it has beat sbwm that aryl hydtocarbon hydroaylase (AHII) is specifically induced in the hrg and kidney of .ninul subjects by the inhala tio. of cigrette smoke, whertas ciprette snwke tus .o ttclioe oaAIIH atlivily ia the liver. These obaervNions were attributed to the roule followed by die cigarette smoke components after inlulatiow. However. the p.per prescntcd here, which deals with two sowes of administration. inhalation and i.p. injection..dds anothn parameter to the investigative pnuess. Whee administered i p., ciRarelle smoke cadcnsale (CSC) and benm(o)pyrene ( BP) in low doses reached the liver before the other organs. Neverdrc- less, thc long was still the most sensitive organ of the three. Resn)1s showed thN, in the liver.nd kidney. basal AHH activity (which is low in the (eqn)'acreaxs rapidly dter birth to reach the adult level two nqnlhs INer, and is only inducibk by CSC and low di»es o/ BP iw unweancd rats In the hrng, however. the basal Allll activity (ow in the (crw) urcrease. .lwlrly al birth. peaks in five-ay old rats and then decreases .IbRMly t'owrti w ewryme acu.uy in other tirsues, lung AHII canrrM be induced in un..cawed y.rnR .nrmal. tAc enryme subuVueMly bccumea sensdrve rn rndurmg a6enu and is IuRhly uwlot sPk in 90 day oW rats Srmrlar behavior has been shown to .W , occur in two other enzymes linked to cylochrome P,ISO: etlwaycoum.rie deedrylase and ethoayresorofin deethylase. According to the wdtors of this paper, the special AH11 inducibility of the lung of the aduM animal is. very important biological fact, and forthcomin6 studies should give insight into the cause and biological consequences of this phenomenon. Van Cantfort, l. and Gielew. l.E. , s.itisA 1o.vrwl aJCance. 4d:9o2-910, 1911. • OAs sapy..t: Fonds de Ia Recherche Scientifique Medicale. Frauw the 1Aboratoire de Chimie Mbdicak. 4witrl de PtlholoRie. Uaversit! de Lifte, LilRe, Belgium. ARYL HYDROCARBON HYDROXYLASE ACT1VfTY IN PULMONARY MACROPHAGES AND BLOOD LYMPIK)C.'Y7ES In the strdy reportod k,ae, the sinwN.wews anNysis of (1) ssyl hydrocrbo. hydroaylre (AIIH) iedretion is ctiltrned lymphecyln and (2) in s/rr levels of AHH activity is pulmonary dveolar nucropbages (PAMs) wr wrkrtake.. The study Rroap included eight Iu.R cancer patieMs with nbeMOs eapoare, 15 .o.<aaioer palic.r wNA asbes/oo espvsrre, 21 priew with lung cancer bM without asbestos tapoawe. ..d 40 patients with .eitDer hrwR c..cer .or sbeaoa eaposwe. All patients were ciRrene snakers who were .d,aiued kr i.dicMio.. of ptrao..ry disease. Wlw .N pnient grorps were compared in terms of their Individual capacities for tiawes to be i.ducad, s•rikinR differences were seen betweee tAe Rrotrps. Those individuaY with r+tEeatos eapowre presented increased AHII activity when compyed to those witho.t asbestos eaposYre whera lung cancer and aon-ea.cer Rroups were compared. Amarg cigarette senken with neither lun= eaucer anr oceetp.Honal asbestoa eaposwe, 6ft h.d telrively low AHH levels in both lyrnplacytes..d PAMs. When AHH values were eaasei.ed is asbeuo.<apaaed snaken widroul ks.R cancer, 57% of the i.dividr- ais eaMibiled k4w AHH levels iw bolh tiasres. When a uewllaeeoos cpep.rison wr t..de for cigarette smoken with lung cancer but withotr asbesws eaposwe, only 7691 o( dr individuals could be clasi6ed as having low AHH vN.es in both lymphocytes and PAMs. FiwWly, when AHH values were compared hr tissues oMair-Ad flom aabeskntaposed hw4c.noa pnieaMS the dMa showed tlut 100'><of these udiviArda poasesstd hiRh AIIH activity i. eidrer lymphocytes ar i. PAMs, bw wa is botb tiswes. Overall, these results suggest thr sdbeswa exposure re.y ef fed changes in a person's AIIN responsivenessor tolal induced kvel is sr.chew.y a a innease tIW iwdiv'd~iul• iul•s s wsceptibility to the materials fornd in cigreue smoke couder.swe. Swodgras, D. R.. Mclamore. T. L., Teague, R. B., Wray, N. P., aed Arsber, O. L. CHEST lOS:12S-/IS, 11181. , OtAri srqpsrt: American Caecer Society. Veterans Admiaistralion Hospital, Hous- bn, rd the National Instmrtes of IkaMh. From the Departmenl of BinkKicd Sciences, Notth Teass S1.u Univenity, Den1o.; tlepntmeM of Medicine, Baylor College of Mtdicine, and the Vdenms Adminirrs- Iio. llospitd, Houswn. I 8 9

M111.TIPI.ICITY ()F CYT(x11ROME P4S01N PRIMARY FETAL IIL'PAl (X.'Y7FS IN CULI URE The pmnnaW period of life is a critical time for the quantitative and qualitative development of marosomal monoosygenases, and earlier observations suggest that fetal hrpaoeytes in cuhure might constitute an ideal lool for studying the perinalal re6ulaatry mechanism o( mowoosytenases. Whik it has been known fa a whrk Ihat Primary fetal ral hepatocytes in c.Nve display difkrenl monoosy`enase activities which can be induced by sevenl chemical indreen. these hep.tocytes were believed wmi now to produce only one single eytodnome P-IS0 species, namely the cyto- chsorne P,450 (ar P4IS). Nowever, i1 now sxms possible b induce othercytachrome P450 species in these hePMocyses. prov1An they receive an appropriate hormo- .al teermenl. in the .rork tepated heae. es.rwinaliar was made of the effect of dearwcMaaone on various mowoosyUenres and on die type of cytochrorne P-I30 wrpporting these enzymk activities. ilree enzymes. aryl hydrocarbon bydrvaylase. ethosycounsrin doethy/ase and aieria suorwoayjalse, were measured (or this pur- po.e. Resrha of this study show IAaI die presewce of des.methnone in the culture Rr.dwm prodrees qudiW ive .nd qswiutive c#.n6es in the Inonooaygenase-srPport- is6 cyrocMane(s) P450. Fa krw desamedwo.e eoncentrstiows. acylochrome P450 is farnmd displaying biochemical yd biophysicr propcnies similar lo those induced by Phewob.rbilal in die adult rat lirer. At higher concenlstlion., similar qualitative changes are observod; but a quansiwive phesomenon occws. the (cylochrwec P4 50)- dependent enzymre s+ctivilics being also induced. Desametlusoae also has a syner`is- 1ic ef(oct in tLe indretion of enzymic activity by the miature of phenobabilal plus benzrafracewe. The various biochemical changes induced by desamethasone in die fetal cell cultures parallel those observed in vivo during the perinatal period of life. 7Aesefore, this eell culture system may cawrlule an interesting model (or studying the ootorsyc development of liver monooaygenases. Kremers. P., Coujo.. F, De Grseve. 1. Van CrwfoA. 1 and GKf.n, /. F. Eavapean lowwal oJ eiorhtrnutry 116 67-72, 1411. OrAar sawart: Fonds de la Rocherche Scieatifique Mldicak. From the Ls6onloire de Chimie Mbdicak, lnstitw de P.tholol~ie. UnivcrsNf de Litse. Lilge. Bdgirwu. DNA METNYLATION IN NORMAL AND SVIO-TRANSFORME() }IUMAN FIBROBLASTS The 5-methylcytosine base eontent of DNA in fow norseal and fow SVIO-trans- forsned hwn.n diploid 6brobldt twkrres was measured by high performance liquid eMorn.a+jraPhy (HPLC) ResuNs show dr1 the percenl o(cytosines methylaled for the (uw nnnnal cell lines ranged from 2.23 to 3.16, whik the range far dre (nur SVIO- transfarrrrd cclt% was 6nrn 2 90 lo 3 03. The mean frx the total number u( IIPLC drrrrmrmuuns w as 2'14 • ll 29451 dclerminalMw+s) frw the mxmal cell types and 3.00 t 11 211 /11 dcrnmmarNrns) lur the Irsns(ormed hnes Thus. in cuntrasl lu other reported studies camparin6 normal and oncoseAically transformed cells, no spp.renl difference was observed in the 3-melhykylosine to cytosine base ratios in the two ceU types. 11 is worth emphasizing that dre IIPLC method used here gives an absolute measure of die DNA bases. In addition, the plrrNy of the DNA is controlled by monitoring for the presence of wacil. Other methods using radioactive label may be hindaod by. various differenl artifacts. Diala, E. S., Pkat, M. M., Coalsan, D. W., and Hoairwn. R. M. Bioc/lrrnicaf awJ eioPAyskal Research Cowrnrnkorions 102(1):U79-13LN, 19%1. OrAer salip..t: National Institutes of Health, The United Cancer Cowscil, Inc.. The Cancer Researcb Coordinating Committee of the University of Cdifarnia, the Aca- demic Senre. Univenily of Cdr(orni.. Sam Die6o, and the Leukemia Society of America. From dse Department of Podiatrics. Univessity of Criforwia at S.n DieRo School of Medicine. La loll. I CONS7TTl171VE BEHAVIOR OF METHIONYL-1RNA SYNTHETASE COMPARED TO REPRESSIBLE BEHAVIOR OF METHIONINE ADFNOSYLTRANSFERASE IN MAMMAUAN CELLS Mcdrionine, becwse of its roles In pralein synthesis and in nsethylrio., b of tentr.l inpartas,ce to a!1 cells. In the metabolic proeeu, aredlioeine can be wed by the oeU Ilrwth two different pathways: (1) methioalle can be eonvesseA so S-adeooo- sybselhionieen die Inyor mnhyl source for eepdsr twmethylatiow reactiown and 1he w.ce of the propyl.mine group for polyanwne biosyndrcsis. or (2) it can be convened to meMriowyl-tRNA, an iaportrM iaesntediae in p.olein biosynlhesis. Is the paper p/eseMed here. il is reported thM anethioeyl-1RNA sysWretase, unlike loethioni.e adenosyhrwferase. beh.ves in a cantilwive mrlner with respect to the concentrsrion of inethioaine iu the culture anedirla. lllis behavior Is see. in Chinese h.nater ovary cells rd in normal diploid and SVIO-trmsforsned brre.n fiboMasss. Although die kin4ics of regulation of arcdliowine adewosyNrawsfer.se and anethionyl-IRNA synlhe- tase by esoUenous mclhiowine are cleady differenl. the levels of the two enzymes hl lhe hwn.n cell lines re simibtt. Rrbnilz, l. E., lacobren. S. 1. and HoA4wan. R. U. Siochiwlca er eioDOiyska Ac1a 677:2d9-273. 19111. OIAai aarr..f: National Instilules of Ne.Nh, The United Cancer Council, Inc.. llro Cancer Reseuch Coordinating CalMnitlce of die University of Califonlia, the Aca- demic Seaue. University of California. San DieRo, and the Lerkemi. Society of America. Prom die fk(wtment of Pediatrics. Universqy of California at San Diego School of Modicine. La Jolla. ' 13 12

F!)I.AII? P(H.Y(iLt1TAMATE AND MON(Xi1.UTAMATE ACCt1Ml1[ ATION IN N()tMAI. ANI) SVIO-TRANSFORMED /II/MAN tIBR()BI.ASTS In the peseM aaempt to ascenain the role of folate pdy6lwamates in cell divi- swrn, it seemed neccssary hrst to surve the cells of (olates and estimate their irMallydNe requirements fnr growth. All fnur ceil lines studied here, the mxmal human drpluid fnreskrn hhn"ast PA, the normal human fetal drPloid AF2, and the SY40-trans- (nrn+ed bncs PS and P1, showed sinwlar kinetics for falate starvatiar. SepFadcs C-10 gel hhratwws chromatography was wed 1o wrcawre the accumulation of fdaoe prlyglu- tamate and nwwwoglwamate in all of these cell lines. After the celis had been depieted of fiAates, they were provided with IinMins aero.nls of (711-fulic acid in order that the cells would accumulate only forms of folale nxessary for proliferation. Bah Ihe a.rnal and the transformed cells accrnwlaled monoglwamate and prly6lwamate forms, hw by 72 hwrs of labehn`, the transformed cells cvuMained 3 K) times nwne p>ty~lwsnwe then the nmmal cells The Rrowlh raes for the rwwmd and IanslrMmed cells were similar at this limiting folic acid eoncewatwrn. Thus, rf k>1Me p4ygluta- nwes we mnee rmpwtw for the prolrkrsiinn of SV141ransformcd cells than the wursnal cells,lhe inMbitiow of polyglwarwre fornsriow could possibly be an impextaM potential trger for chenwuherapy. Ho,/fiwaw- R M to ol lewwol nJCrllolar PAysioloRy 109:497-503, 1951. OUAn sayprr: Narwsrrd Instnwn of He.hh, The United Cancer Council, Inc., The Cancer Research ('oordrnatrng Commmee of the Unlversily of Cahforria, the Aca- demic Senate, University of Cdrfornia, Sam Diego, and Ihe Leukemia Society of Amerrca. ~ From the DepartmeM of PedrMrics. Un/venily of California N San Diego School of Medicrne. La loll., and the (ienetres Unit, CMldren's Servrce, Massachusetts (kneral Ihnpital, Departmenl of Pediatrics and Center for Human Genetics. Harvard Medical School. Boswn. DNA METHYLATION IEVEtS IN NORMAL AND CHEMICALLY 1RANSFORMFA MOUSE 3T3 CELLS TMs {nvestiRation was undertaken 1o assess the effect of chemical transf.xnution on tntal genomic DNA methylatwn as measured by high p:riormance liyuid chnwna- aWq+hy IIIPI.C). In the study presenled here• norsoal mnuse embryo l7l cell cultures nrwl dwse oncoRenw:ally transformed by the chemical carcinosens benrola Ipyrene and methykholanthrene were andyred by HPt.C 1n determine the S-me11 ycytosine to cytosine base ratw» in their total genomic DNA. Results showed that thc mean fur 10 11 W.C detersniraliaa of the normal n) cells was 2.87% of eytosines melhylated with a standa.d deviation of s0.Se, while thM for the benzola)pyrene-transformeJ 3T3 cr lls and the mnhykMrlaMhtene transformed 3T3 cells was 2 99% ± 0 N(11 dclermi- n.u..n.l rw1 2 NI's •// IN IIS ikrermreatwwxl, resptctrvely llrcu reudts led to the i.ww lu.i..n rhr itwic rt rw- rrd ddlcrcnce rn the e.rrnt of wwal Ecmr.na• I:NA mrthyla- r..o t.F..«n mrnul .rd. hrmw all) uan.lown.rd 11 1 icll.,rhen mc..wcd hy 1/1M C I Diala, E. S. and llroffnrsn, R. M. ' Aioclkrnr,nl mwreiuqrAysicnf Resrur.AConunwricwiuns 104(I):11d9 1491, 1982. OrMer rrhert: Natinna11ns1itules of Heahh, The United Cancer Council. Inc., The Cancer Re-earch ('.rsrdinatin6 Conunetee of the Univenity of ('altfomia, 1he Aca- demic Senate, University of Cdifarna, San Die6o, and the Leukemia Society of America. Fnwn the Department of Pediatrics. Univer%ity of California M San Diego School of Medirine, 1a lr+lla. MF.TIIIONINE [W.Pl:NI*-NCE IN CANCER CEI1S-A REVIEW Methirmine dep:ndence occurs in a large rwmber and wide variety of cancer cdb a.d dnes arw seem to be a r.ndorn eomponenl of the 4tnsforwted phenotype. De6nitio. of methionine dependence slales thM it is a defect found in many cancer eell liwa 1hM inhiMts their growth in culture when methinaine is replaced by its immediate prec.r- rx, hurnorysteine, in the culture medium. Norsaal c'uAwed cells do no1 have this defect. This report lists the diverse and large nurnber of animd and human cancer lines thr we methionine-dependent, and critically reviews the cell brotoRy ud meMioeine bioehemisuy of the plKwoarenon. Haf/iwaa. R. M. In VMre 1i(3):121-12f, 19R2. Otlrer sappert: National lnstitwes of Health. The l/nNed Cancer Counell• Inc., Tlr Cancer Research Coordi.riwR Committee of 1Ae University of C.fi(orui., Ihe Aca- demic Senate. University of California. Sr Diego. and the Leukemia Society of America. From the Department of Pediatrics. University of California N San Diego School of Medicine. La lolla. HYP(NMETItY1.ATION OF IIFLA CEI.t. DNA ANl) THE ABSENCE OF S MI:1'11Y1.CYTOSINE IN SVIO AND ADENOVIRUS (TYPE 2) DNA: ANALYSIS BY IIPLC in the study presented here, methylation of 1he prihed virion DNA of bnth SVIO and adenovirvs /typ: 2/ was mcarired by high perforsnance liquid chromatography (IIPir) and compared atheir Mrsts, African green monkey kidney cells and HcLa ceNs, respectively. In SVIO DNA, as much as 12 nanomoles of cytosiee has eeen measured without cwrcomitad delcayion of mCyt. SVMI contains 27 Cp(3 pain. which is the usual methylatam site. If all Cp( j pin were methyla/ed, this would yiekd a siRnrficant 1. )'16 methylatiun of L'tal cyhnines; however, in the virion 1)NA studiod here, nmwa scemed to he methylaed Also, as with SVN), mY'yt was nnt peseM in the IS It

.1 1 IN the com-lusion that candage contains estraclabk matrss compounds that inhibit inva- sM"n tn an eapcnmeMal system When these dilfusahk and estractahk sulnt.nces were lunhcr studied. it was found that the rnhibtian of osausarc.wna cell prt IderalirMt was caused by molecules with a molecular weight of less than 50.4100 daAtins 1KGsm this anu invasive facair of the cartilage estract, a pruatease inhihitur was tdaNilKd thal has the aMtbty to inhibit mamalun collagenase. including that elalxxaled by ostunarcoma cells and endothclial cells These esperimental data led to the hy(w+thesis that invasion cof cither tunrir nr endcwhrlul cells depetds on praeulytic /a+llagemdyticl enzyme n tivNrcs Other studies aktns these lines have been instituted to follow the invasive- ncss and prohferation of bladder eancer. Kurnrwr. K. E. and Pauli, B. U. In Gdbcrt, H A, Weiss, l.. and Monsew, D C. G. (eds. )' Aowr Mctouotn, Boston: (: K Hall Medical Publishen, 19l1, pp. 1)1-163. Other sayla.rt: National lnswitutes of Ikahh. From the Departments n( Orshopedie Surgery. Paitholosy and Biochemistry. Rush- Presbyterian-St. Lute's Medical Ceneer. Chicago. ANTIINVASION FACTOR MEDIATES AVASCULARITY OF HYALINE CARTILAGE To test an arMiinvasion factor IAIF) hypothesis of the resistance of cartilar to vasculr invasion, a novel in vitro system was studied Iha1 employed bovine .rttcular carttla6e aa a growth surface (ot normal heparin-ssimulaWed eaddhelial celis. In this study, cells were tested for their ability to invade the malns of viabk and devitalized earacted cartilage as mrwutortd by thin-seclion electron microscopy. The growth behavior of cells on devitalized estrsetod cartilage was esnnined in the presence ard abaence of cartilate-derivedc estrsclable AIF in the culture medium. Whereas wur- ' mally viable att/cular cartilage is a poor growth surface for Cndolhehal cells, the eadnthclial cells studiod here. is eataasl. grew as conlact-itJtibited anomslayers of Ilaaenod cetls on the surfaces o( estracled cartilage. The cells were separated frorn the carulage tnauis by abwtdant bwl lami.a. There were a few micnoviUi at the basal ptasma teembra.e, but there was no de:Sradatioa or penetration of the eolla6enous snnris of eatrat:lod cartilage. Hosvever, wAa e.dashdid cells were stimulated by hepari., they assumed a polyheAnl sh.pe and Pc^tlr>NeJ the estracscd cartilage with awnncroas microvilli and some eyloplasmic processes. 'tlws penetration of Me colla- <enous matru was associated with tlswe rarefaction a.d degradation of eollagett fibea. heportamly, however,llus invasion of hepari.-stimulated endcMhelial cells was .bdished when (ow conceatr.tiom o(e.nilaje-dcrived AIF were addcd to the culture medium. These data provide evidence thN the resistance of hyalame cartilage to ee- dadhelial ccll invasion is resulaled i. patt by tissue derived pnNcinase inhibipxs and a. sntipmlikrMive activity directed atainst endolhelial cells. Ku.untr. K E. rt o/ Seiwinors in ArtMftis A RAera.otiun 11:67-69, 1951. Other sr.*pr1: National Institutes of Hedth. fnwn Rush Medical College. Chicago. [E I C1IARACTFRIZaT10N OF ADULT BOVINE ARTICULAR CHONDROCYiES IN CULTURE AAiculr cartilage slices, obtained (rom IR-naold bovine me/acarpnphda.beal joi.ts, were used as sowce mNerial for thia descriptive article of chatdrocyla in culture. After sequential digesliowt s.d 6hrMioe, cells were plated in either tissue culture dishes or roller bottles. Chondrocytes f ned iu buffered glulraldehyde corai.- ittR 0.1S svthenium sed, were esamined by Ii`M and transmission electron micro scopy. Collagen type deknrtin.tipt of 'H-proline-labekd proteins isolated from cultures were performed by elecyrophonyic atd CNBr peptidt: analysis. Biosymhesis of praeoillycans was measured by »SO, iwcarporatiom inso wwromolecuks eslractod under dissocialive conditions. Eaatei.aion sMwed qu1 isolakd clwndrocyles prior b culture were typically r.otrried with scaM tetriwrid mrris, which could be Rmoved by mild trypswrzation. TMatghout IMe prognession o( drc euNures, phenotypic .heq- 1iow. wete .ot obser.rod. E(pclro/luorop.plp of eo11" thal was entracsed from'N- pmline-labekd cuawes aflet nwW PeVundigesoionshowed i band i. drc positio. of the al chain; an e2 ch.in caad rwl be delccted. Cya.oRem bbaeide peptide analysis oblaiaiod 6wn type 1 mlor radioacyive peptides eo.wirabd with unlabeled pcptides ype collqe.;lype I eollagert was taot delecublc is these cultures. PsuseoglytM aggregate was eatracted fran both culture disAes aad mlkr bottle cul- lures, under associrive conditions. Thpe wem diffese.ces eoted in this esperannr suggesting that tfrc roller e.Uure eca-associaled ananis may have a pener depee o( organization tlun that pows in su.0ard tissue culture dishes. These same data also indicate that anicular dqndtocyles Rrows is mas ro(kr cuhures we capable of syathe- sizi.6 a phcnotypically stabk, tissue-like nulri: in vitro. Xartawr, K. E. ed at. Se.wawrs iw ArtArit/s t RAnreoHaw 11:101-10), 19d1. Other npport: Nriow.llawitutes of Heddt. From Rush Medicd College and tlte U.ivenily of Illinois Deet.l Schoo(, Chicago. REGUTATIUN OF TUMOR INVASION BY CARTIUIGE-DERIVED ANTI-INVASION FACTOR IN VITRO Manunaliam crtilaV is highly resisunt to invasion by tumor cells. This asiss- ssroe was swdied here wi/h 1be use o( a oovet Jn vlno culswe system. Aniculr catilade obuined rsan fresh metacarpophdanteal joints o( preadolesceM bovines was used n a growth surface for huma. TE-IIS osuosaRonu celh and foreskin flbrohlass. Cartilage d'ats formed the boltoms of sui.kss-wteel cylinden, providing closed growth chanm- ben Gor these cells. Both invasive osKOSUroma cells and normal FlrroMasrs were tr.abk to pennr.ne viabk, uneatracad catilage durinR a 2-weet cuNwe peaiod Whe. canibge was devitalized by freezing and duwiqg, die tissue remained resistant b invasion. Cartila6e, eslracled with either I or 3 M tuanidine hydrochloride, was invaded by oueoaarcoma cells, but mA by control fibroblasts. Invasion by osleo.r- eana cells into saN eatrae/ed eartila6e was abolishod whea low concentrations of a cartila`ederived, atqi-invaswon factor were added to the culture medium. These dNa 19 ,r

_L provided evidence Ihallhe resistance of cartila6e to Iumrx invasion is regulated in part by tissue-rkrived pr.wemase inhibAon. Pauli, B U., Memoh, V A and Kuetrnrr. K. E. hwrnof oJ1Ae Nurr.NSOI Cone er lestitute 67(I):6S•77, 1991. (kAer r.yw.t: National Cancer Institure. FnNn the f)cpar(ments of Pathnlogy. Biochemistry and Orthopedic Sursery, Rush Medical College. Rush-Presbyleria.-SI. Luke'a Medical Center, ChicaSr,. IN VfTR() DETERMINATION ()F TUMOR INVASIVENESS USING f:X1RA('TI:D I/YAt.INf: CARTILAGE In this anenspl to determine whether sah<atracted cartilage could be u sed as a tesr connective liuue for in vitro discriminaion between noninvasive and invuive tumor cell hna, a novel in vitro nsethod wY devised which used salt<suacled, bovine nucular carlilase as aa esperimeaAal Rrvwds surface /or both normal bladder epithelial cells and rwiwinvasive, invasrve, and mefastalic carcinoma cell lines derived fnrm chemical carcinogen induced tumors of dse ra1 urin.ry bladder. As monitored by Ihin- aection electron microscopy, sah<atracsod cartilage was readdy penetrated by the invasive and nrUstatic rat Madder carcinoma eell lines. The metasrurc cell line could be differentiated from the invasrve, noametauric cell line by its grea.et depth of InvasNM In ccrwrast, noninvasive carcinoma cells as well as normal blrkkr epslhehal cells lacked the capacity to crrtsde and penetnte the eatracled mtlris of the articulr cartilage IIsrn6lhesedefinedcell lines,sah-ealrnctedcartda6ecanbeusedtorepaxlu- cibly discriminate between can uamas having different invasive polentials. This assay system may have diagnostic application for the tn vitro staging of lumura. Pauli, B. U., Memoli, V. A. and Kr.rtnwr, K. E. Cancer Research 11:2061•2091, 1981. OOther sayl.rt: National Institutes of HeahA. Froru the Departments of PaUsobffly. Biochemistry, and Orthopedic Sur6ery, Rush Medical College and Rush College of Health Sciences, Rush-Presbyterian-St. Luke's Medical Cenrr, Chicago. A FAMILIAL AGGREGATION OF PANCREATIC CANCER: AN IN VITRO S7vDY Pancreatic cancer, with its olncwe etiology and diffscuh early diagnosis, presents a medkal problem of slaggenng propurtions. In the in vitro study presented here, a family was identshed in which four individuals manifested pancreatic cancer verilied through two generations Cell cultures from split-thickness skin biopsy specimens were nblauied frr 24 rnembers fnsm three jenetNions (17 bMrdlirse relatives, seven family memBen by inarruge) as well as len noefamily normal subjects, none with a lamily hwory of aild sunrxs One of the conslam features of human nwNwolayer dcrmal (ultunrs has been dipN»dy On the other hand. hypenhploNly. other than 1et..pkidy. has been rarely observed in cultures fmrn.ormal subjccts without a family history of solid tunwus. In this shdy, however, increased in vrtru hyperdiploidy was observed in eight of the 17 family members /esled. In the long run. allregates of pancreatic cancer in families such as this one. and the occurrence of pancreatic cancer in sonse .wosornal domiarst cancer syndromes have added credence to the relevancy of agenetic component(s) in a fnctioa of patscreMic eancers. ln an important way, this study has demonstrated the potential lnr eombirdng detailed family data with rocogni- lion of in vitro biornarters for caucer pronencss an am approach ao rhe cornpehension ol carcinopeesis in pancreatic caecer. D.nes, B. S. and LyncA. H. T. IAMA 217(20):2798-21102, 1982. Od.r nff.rs: Naliond lawiluks of Heahfi, Danes Medical Reaeandt Fu.d, Cor.cll lhdveairy Medical Cdkge, and ?.emwray Foundation. Fruas Ihe Laboratory for Ce11 Biology, Department of MedKine, Cornell University College. New York, and the Department of Pmventive Modicine/Public IleaMh, Creighlo. University School of Medicine, Omaha. GENETIC/EPIDEJNIOIAGICAL FINDINGS IN A STUDY OF SM(NCING•ASSOCIATED TUMORS In dsis Rendiclepiderniobgical sludy family histories of cancer were obuinod via personal interviews frvre eonsecMively.scertained eancer patients who were under evaluation in one of two University Oncology Cli.ics in Nebraska. Inchrded is the srriea were 147 breast canetr pobands, 85 eolor cancer proe.nds, U lung c..ca probards. and 111 prahnds with other casmen Yot have been n:patcd b Ee asocired with cisarelre smoking (carci.orna of tlse oral cavily, esophaRtts, p.aereas and uriaarp bladder). Smoking histories of pwbands and their relarives were obtained for an oveslappinR series of 60 hrtg ea.ces prob.nds and 7111 psnbands with other smokiy- wociarod romors. Findi.p revealed tlw, aNho.lllt a significam cohort effect was observed with respect to smoking habils for bah relalives of lung cancw probands.nd for relarives of probanda wilh other smokinR-associMed rwnors, a corresponding arewA far hsnt cancer frequeney was observed only for relatives of lung cancer probar,ds. This resuN suggests the importance of host faclon in consbinalion wNh environme.ul eaposures in determining hsng risk. A cohun treed for hrsg cancer was also appareM amons reftlives of breast cancer prolwsds, but not for relatives of colon caarer pro• bands, suggesting the posublNy of an intrlnsic association belween carcinoms of the baast and lung. It seems reasonable that further ehrcidaliun of hosllacqor suseepribil- iry in hmg cancer nsay have importantetiolu6ical and preventive implications. LyncA. H. T. et at. Cancer Genetics and Cytosenrtirt 6:16)• 169, 1952. FrMrs the Inslilule lor Familial ('ancer Manasemenl and Control, Inc.. Departments of Prevenrive MedicinetPuMic Ikalth, Surgery, and Pathology. CreigMon University Schkal of Medicine. (Mssaha. 20 1 21

LOW SERUM ISA IN A FAMILIAL OVARIAN CAN(-!:R A(GRF.GATE Ahhrw6h familial ovarian cancer is increasingly recognized. consutcnl biomarker associalMms that correlNe with its risk have remained elusive In thrx pq+er, however, a family is described lhat is characterized by escessive occurrences of ovarian carcinnmw transmitted in a pulere tronsistent with an aulosomal dominant factor. This family is unique in that identical twin sisten each had verified ovarian cancer and each had a daughter with the same ksion. Upon familial investi6:.Iwus,11 of 45 individuals tested has serum ISA kreb below the 95% range. Careful consideration of these results showed dw low aersr. IgA levels were found to segregate in a sufficient number o( individuals from dis farwily, /o suggest thal this may be a 6eneli- cally determined immune defecl etioloSitally Integral to cancer susceptibility. While dvt pxarrve role of IgA in pdhogewesis rewuiws elusive still. N seems apSwenl Ihal cancer•psne families should be thorvughly investigaed for further elucnWion of these phemmrena Schuelke, G. S., t.yncA, H. T, Lysxi,l. F., Fai., P. R., and Chaperon. E A. Cancer Genetics and Cylo6enelKs 6:2)1:2)6, 19e2. Other sarya.f: Elsa U. Pardee Foundation. From the fkpartments of Medical Microbiology and Preventive Medicine/Public /kahh, Crci6hton University School of Medreine. Omaha. NATL/RAL IIIST()RY OF III:REDITARY CANCER OF TIIE BREAST AND COLON Anecdotal repwts have suggested that survival characteristics of hereditary fnrrns of cancer may differ fmm their sp.adic enunterparts. To test this pnsrhdily, a review o(disease free survival was undenaken by evaluating the affected members of heredi- tary colon and breast cancer families within the eslensive Creighton llnivessity Oncol- ogy Ctnter familial cancer resources In the study presented here, the natural history of 106 patients from 1 d families manifesting hereditary breast cancer syndn mes and of 117 affected patients from 20 families manifesting nonpolyposis herednar) colon can- ea was evaluated Findings were compared with the American Cdkae of Surgeons (ACS) long-Ierm audits for breast and colon cancer respectively. The cardinal features of hereditary cancer were observed within the study troup, inchdin6:/ I) a sianificant youn`er, aage of onset (49 yean, brtast; 46 yean, colon); (2) an escess of Prosim.l ksions in the hereditary toMsn series (19116); rd (3) an eacess of bdaterality in the hesodilary breast cancer patients. The clinical sute at presentation was similar for the hereditary rd ACS audit pMieMs. Five-yer survival was significantly impmvcd for both herafiury eancer populrions as eompred to the ACS wdits (674s% hereditary breast cancer awd S2'R nonpolyposis herodiury colon cancer). Improved survival in herediury eolon and breast cancer pMients may h.ve a bearing on the design of futurc clinical protocols. AR+.no, W. A.. Recabnen.l. A.. Lynch. H. T.. Campbell. A. S., Mailliard, J. A., Oreaw. C. /1., Lynch. J F, and Kimberting. W. J. Ca.rer N1(2) )6() 1d9. 1982 Oz4r ryP.rr. Nrrnul Cancer Insuqute Fnun rM Inssrrwc Irw famdhsl Cancer Management and Conlnd, Inc ,('rerghkm l/mver.ay k Iwrl ul Mede rne. l hnaha Nl:()1'I.ASTK' TRANSFORMATION OF RABBIT CEl-tS BY MURINE SARCOMA VIRUSES Whik rabbit cells have been widely used for isolation and replication of a variety uf rarovirusca, Ihey have sekkrm been used for Iransformaliun studies with rmrirse sarcrxna viruses (MSV). In this prper, hnwever, neoplaslic transforsssNion of rabbit cells by Kasten MSV (Ki-MSV), the Ki-MSV pseudolyp: of baboon endo`enan virus (Ki-MSV lHa1:V/) and the Mrrknsey-MSV PseudnlyFe of feline kukemia virus (M-MSV jFc1.V /) is rcprrted Rabbit cells can be readily transformed by KiMSV, Ki- MSVI H.EV ) anJ M-MS V( FeI.V ). Rabbit cells transforsned by Ki-MSV and M-MSV- (Fe1.V) were found to be virus poducen, whereas those traesforsncd by Ki-MSV(BsEV) were wonproducen (NP). The NP cells were obtained by simply infecting rabbit cells with Ki-MSV(HaEV) and subesdturing the infected cells. AI- dwwgh the nrnpholo`ically aitered NPcefls did wa produce infectious virus or murine leukemia v'aus aetisen, they did contain a rescuable MSV genome. All of the lrrss- formed cells frwmed c.rlrsnks in soft agr. grew b Aigh saturation densities and po duced tunuxs when transplanted into nude mice. This report is the first one of successful tumer induction by virus-Iransforsned rabbit cells. The ability to produce sarcomas in rabbits by Ki-MSV and M-MSVIFeLV)-transfamsed cells should provide  useful additional model for studying ehemolinrrawrotherapyy as well as imnnwso- pevention of rabbit cancers. Rhim,1. S., Bediguu, N. G. and Fos, R. R. (Meier, H.) Mrernoriowollournal oJCancrr 30:365-369, 19112. Other sq/.rt: National Science Faundalion and the National Institutes of Health. Frnr,s the /AIxxatory of Cellular and Molecular Biolo=y, National Caacer Institwe, Bethesda. MO, and The Jackson hboratory, Bar Hasbor, ME. a 11. The Respiratory System ELASTIN BIOSYNT1If:SIS IN CNICK EMBRYONIC LUNG TISSUE. COMPARISON TO C/IICK AORTIC ELASTIN In this biochemical study, the synthesis of elastita was followed iw chick embry- onic lung and compared to that seen in embryonic aurtic tissue. Messenger ribonuckie acid (mRNA) was isolaled fn>rn both hrnS and aottic tissues and translated in an mRNA-dependent rabbit reticuluryle lysate. The rewhs densonsune dsal both tiswe RNA preparations direct the synthesis of two elaslin pmleins possessing molecular weights of 70 41110 and 73 (KMI, which are immunupetipitabk with anlibody directed against chick aortK• Iruprelastin I)rgae cuhure of embryonic lungs and wAas followed by eatraclNm of the ('t1/valine labekd proteins with urea in the presence of reducing 22 1 23 .

SYNTHI:TIC Ia.ASTASI: INIiI81TORS AND THEIR R(X.E IN THF. TRI:ATMI:NT 01; 1)ISEASI: Pn1leaxs (pnwcrn hydrolyring enrymesl arc involved in many inrpnrtant natural bir*igicsl pmccsscs: they are also,htw6ht to be involved in many diffcrent diseases. ()ne suk h disease is pulmemary emphysem., which seems to result from an imbalance bctwetn proNeases rrkased fnwn human Icucocytes and the prdeases' inhihilors. Since much evidence has accumulated showing that ptoleolysis of lung elastin kads to the devek,pment of emphyscma. i1 seems evidenl that selective elasuse inhibiton could be used in the trealnmcnt of this diseae. The inhibilors used could be natural inhitMtors such as a,-praleinasc inhibitor itself isolMOd itao biood fractionation, nr they could he synthetic materials. Wrxk done in the invesliplots' lahorawry ar date has shuwn that synthetic elastase inhibitars have considerable polential for the trcatment of emphy- sema. Two pcprde chksmrnNhyl kelone elaslase inhibit.xs Ac•A7a-Ala-Pro- Ala('H,CI aedSuc Ala•Pro•Va1C11,C1 havebeenshow.losigmAcawrlyd~rminishthe esrc./ of esperimental<lastne•iwauced emphy.eme in hannten. Me(I Suc•AIa Ala- Pro Va1C11,C1 has been shown 1o be orally active is providing pntcctinn against induced emphysema in r.ts Whik there are s1iN Queations about whether such reactive a1kylMing agents could be used i. the treatment of emphysema in man, the animal studies have shown that claslase inhibitors can be used to treat emphysema, ThercfG>,e, based on these audres. it seems thn the overall prospecls for the deveMopment of a synthetic elastase inhibitor for use in humans in the near future we quite tosd. Powen, I C. rt of (Trovis, l.) In: Rich. D H rd Gross. E(eds ): PrptJdrs: synthesis - structure - Junrrion. pro- rrrdinRs oJrAr srrrnrA Awrrriron pcptidt tyn,posirrw, Rockford. IL: Pien-e Chemical Comp.ny, 1" i, pp; 191. 199 OtAtr srpp: National Institutes of Ikahh From the School of Chemistry. (:e«gu Institute of Technulnsy, Allarua PRO7F.OLYTIC ENZYMES AND THEIR ACTIVE-SIIE-SPECIFIC INHIBITORS: ROIE IN THE TREATMENT OF DISEASE Proleases (prolein-hydrnlyzing enzyrnn or Inotedytic enzymes; have been known for ova 100 yesrs since trypin was firsl isolred from pancrc,Mic juice by KiNne. Fur most of the perird since their discovery, praleases were thuu6ht to be involved only in digestion. Ilrrwever, in the lasl decsde, proleases have bbxn shown to be involved in many other impottanl physiological pmcesses, such as fertilization, cos6ula/inn, and the immune response. Outside of their normal envinamcnl, pro- kases can be estremely destructive, but natwal human plasma inhibitors inhibit most pnneases that escape. Imbalance in pnNesse-pnNease inhibitor systems :an lead Ns a number of diseases of which pulmonary emphysema is one well-charact•vired esam- pk. This disease results when the protene elastase attacks elaslin. the mapK elastic prswem rn the lung ('rN+sakrable effort has been devoted to the synthesis of inhibihws rd pnMenlytic enzymes such as elaslase for possible Iheiapeulic use In the future, spccthc and sukcnve synihcut prrwea.e inhibu<ws should he useful f(x trcaun6 specdic dnca.rs rhal ranRc /nmt the curnnwm cold lo chamre drsarders such as emphysema a h Powers, l. C. (Travis. J.) Ia: Feenay, R. E. and Whitaker. l. R. (eds.): Advoncrs in Chemistry Srrirs, No. 198. ModiJG-otion oJProtrins, Washin6ton, D C.: American Cliemical Society. 1952, pp. 347-367. Or4r ssqprf: National Institutes of Health. Ftom the School of Chemistry. Georgia Institute of Tech.ology, Atlanta. SPECIt1CITY AND REAC7IVITY OF HUMAN LEUKOCYTE ELASTASE, PORCINE PANCREATIC ELASTASE. HUMAN GRANULOCYTE CATNEPSIN a, AND BOVINE PANCREATIC CHYMOTRYPSIN WI7N ARYLSUI.f`ONYL R.IIORIDF.S: t>1SCOVERY OF A NEW SERIES OF POTENT AND SPECIFIC IItREVERSIBIE FLASTASE INHIBITORS The discovery of some potent and specific inhi6ilas for poreiae pa.creaiie (PP) elaslase, human kukocyte (IIL) elaMase, and chymotrypsi. Aa is (eported hae. 1n paAicrlar, 1he reactivity and specificity of a suin of suhslituled benzenesulfo.yl /bmide with HL ela.tasc, e.drepsi.0, PP elwase, and bovine chymotrypsi. Aa+ re desnihod. Benrsnewlfonyl 0uorides with 2-/luoroatyl tubstitueMs were found 1o be poter and specific inhiEiwrs of eln(ne. HL elastre was knhibi(ed nwsl r.piAly by 2-(CF;CFi(`ONII)-C,H.SO,F. PP elwre was twost npidty Inhibited by 2-(CFiCONH)-C,H.SO,F. The 2-(CF,iCFiCF,CONH) and 2-(CFANH) de- rivrives were quite selective for HL ek+stase ad i.hi1ited PP elswse, cwhepsin O, and dtymolrypsi. Aa quite slowly. A specific and palatt ehymoaypsi. inhibilor (2-(Z-GIy-NIl)-C,H,SO,F) wr also discovered. I. this paper. a model for the data.e iahibition reaction is proposed which involves itweraction of the A.otorryl pwp of 1he inhibiwr with dte prwnry wbslrale recognitioa site S, of the enzyme. Overall, drc resuhs of dus study demowstrak dul it is practically possible to eonstrrct simple or`aweic mokeuks which sse specilk idrbiioo of HL elas(ase, PP el.uase, a chymotrypaia. Ya.hirewra. T. er of. (Trods, /.) The Jawnol of Biological Citrntisrry 237(9):3077-3061, 19'2. From the Schooi of Clrcmistry. Georgia lns(itule of Technology. Atlanta. P(MMAT1ON OF A STABLE COMPI.EX BETWEEN HUMAN PROELASTASE 2 AND IIUMAN o; PROTEASE INHIBITOR The studies reported here were aimed at clarifying the s.mre of the a,-prdea.e inhibitor (PI)-bound imnnmoreactive elasuse 2 in normal plasau by investigating the interaction of praelastase 2 with plasma in vitro. Results showed that the major pro- elasuse 2 binding (act.x in human plasma is a,• Pt. and thu proelastase 2 reacts directly with a,-PI via a partid active swe, in a manner similar to that of active eadupeptida.es. When prvelastase 2 was incubNed for 16 h aY 2S'C with 0 1 M diisopTapylfluornphos- phMe, 0.8 mol of inhibitor was incurporated/mo1 of zymogen. The product no longer 26 1 27

capacity ITI(-) lo elivase inhiba.ry capatNy 1 EIC) of o,-PI in these tissues The TIC/ Ek'r.nwr is skM intlucnced by the other paleins in serum nor by the canren;ratrm of a; PI. When this technpue was adapted so measure the popurtiow+of osidited a,-PI in the serum of young aduN, healthy smokers and nonsmokers. 23% oxidized inhtbitur was fuund in the smuken' sen, wheRU no osidi:cd a; PI was dctectabk in sera of arnsnadcn T1ws smnk in6 appeared so lead to osidalion damage to circulating a,-Pl. Re.htctMM+ of the I:IC of the a,-P) in the sesvrn of smnkers was compensated by a I.13- fold increase in lheir serum a,-P11i1e.s. This assay for oxidized a,-PI may be useful in studas of dte relationship between osidarion of a; Pt and the deve1opment of plmo- nary emphysema Besny, K., Robertie, P., Senior, R. M., and Troris. J. T11t hwrnel oJ Iaboratory, d.J Cfinkd Mtlitint 10012):1116-192, 1982. (hArr s.qp.rl: National Institutes of /kaldt. Fro.n the (kprtmenl o( Biochemistry. Uarvenity of Gcortia. Athens, and The tk- partnienl of Medicine, Washin" Universily School of MedKine al The Jewish Ilospitd of St. Louis, St. Louis. ALPIIA,-PROTF.INASE INHIBITOR IS MORE SENSITIVE TO INACTIVATKNd BY CIGARETTE SMOKE TNAN IS LEUKOCYTE I1JISTASE In these comparative studies, aqueous solutions of ps phase cigarette smoke were incubated with p+rt human kukocyle elastase or with ende human kukocyte granule e.tract. nd the effects on enzyme activity wert delersnaKd usinta synthetie .mide subatrMe. Simuh.neonly, the srne smoke solutions were incubated with IOW human serum under identical coaditions. srd the effocts on serum inhibiti+n of prified or crude kvkocyte elastase wne similarly, mmeasured. The in wero resuhs showed that serun elastaae inhibitinL capacity (a,-P1) is mae suscepibk to inactivation by ciga- Rlte smoke dn, is graa ufocyle elastase, when these prolenn ue i.cubased with wNer- sr+lubue smoke estrsns for relrively short tpnes. Similar teesuks were obtained with both synthetk (amide) and natural (ektsti.) wMaNes, gas phase smoke and unGac- tiunated whok smoke, anJ with pure enzyme and ctude kukocyse granule extract. Also, aqueous solutions of unfndionated ciprette smoke were incubated with kuko- qne elastase or seruta, and the abilities of the smoke-treated enzyme to digest elastin and of the srnoke•Irested serum a i.hibit elastin digestion were dctcrmined. Both esperimental protocols showed thM seswn elastase-inhibitin6 capacity Iprimarily caused by a; PI) is rnore susceptible to iwuclivation by aqueous solutions of cigarette smokc than is kukocyte elastase, sua6stirt6 that elastase inhibition (rathcY than elastase . activity) may be predominantly suppressed by cigarette smoke idulat on in riro. /awoD'. A. and fkarint. R. American Rtvirw aJRtrpirarory Distast /260):691691, 1982. (hAtr.rh.rr. U S. Public /kdth Service Imwn the Iselwrnrm u/ 1',aIr.logy. State Urtmvcnuy of New York au Stuny Bruuk, ENZYMATIC OXIDATION OF ALPHA- I -PROTEINASE INHIBITOR IN ABNORMAL TISSUE TURNOVER This review paper describes ptsem knowledge with regard to the interaction of alPha-l-pdcinase inhibitor (a,-PI) and myeknperonidase in rirro. using both puiftod cnzyme and pAymowphawckar kukocytcs as study souces. In the first place. a,-PI was found so contain a methionyl residue a1 i1s readive cenler which was sensilive to oxidation by either N-chkxosuccinimide or by enzymatic oaidMiuw. Chemical o.ida- tiow revealed thM two of the eight rnethio.yl residues present in the molecule were osidrzod, one of which was at the reactive cenler. Subseque.tly, it was found that the enzymatic oxidation by myeloperosidase rran neulrophil `rarwh:s, M the presence of hydrogen peroxide and halide iow, could also be easily Muined. Since mycloperox- idase is present in the same grswks which contain proleolytie en:rmes. it would be expected that both types of en:ymes would be tekased sinwlta.eously during either Ohagocyloais or cell twwover. h is worth noting dtM the rayelapemaidase system does not osiM:e a inaclivre nwsl other proacinae inhisilors or proteMases. In the second ~ place, it was shows rncen/ly that when a; PI was eapssod 1o viabk neutrophils in the presence of chloride ion ad phorbol myrislyl atetMe, the abiMiy of a; PI to bind eksstase was diminished. However. when myckipero.idase-deGcient ceRs or those horn patients with chronic 6rrwlomatun disease were tned,lhere was so effect on a; PI activity, indicating 1he involvement of myeloperoaidase and hydrogen peroxide in a; Pt inactivation. More recenlly, oxidized a,-PI has beett isolated fran rUcumaoid qrnwrial fluid of arthritic patients aed also fran hrnR lavage fluidsof smokers. Appna- inuuly four methionyl residues were fouad to be oxidized in either case. compared to two oxidized residues when ntlive a; PI was lreated with either chemical or e.zymYi- caBy produced oaidaMs. The itdicMinn is, tht:refotee,llw oxidative inactivalion of a; P1 does occur in vivo. Otlri studies discussed here deal with the importance of oaidnioa aemphysema developataN, drc actioru of the myelopetotidne systeet, a.d the possible rok of rnacrophftes is doe osidation o( a; PI. Mnheson, N. R., lanoQ, A. and Trerit. /. Molnrrfor m.d Ctllrfar QiorAendsny 13:61-71, 1912. From the Departmenl of Biochernistry. University of Geargia, Athens. and the Depart- asnt of Pathology. State University of New Yat at Sany Brook, Stony Brook. I RAPID CONVERSION OF.ANGIOTENSIN I TO ANGIOTENSIN It BY NEl7fROP11Q- ANI) MAST CELL PROTEINASES Ansiaensin II is a pep(ide that has ph(ent vasoconstriclor and aldosterone secre- tion activities and is derived from the larger pveM protein an6iotensiro6en, which circuWes in the Pluma. On 1he other hand. angioteasin I is a pcplide that has no significant biological activity md requires further conversion b anSiwensin II. in the studies rtprsed here, so is shown that human neutruPhil cathepsin (: and human skin asast cell chymax rapidly convert an6idensfn I to utgiutensin (1 with only minor ckavage elsewhere in the molecule The rate of cleavage is consistent with a ptential rok .iw eilher ox both of these enzymes in an altereate puhway, for angiwensin 11 MI 1 31

. synthesis. Since neither enzyme is inhibited by caplopril, an an6iotensin converting enzyme inactivror, i1 is Possibk IhM lerkocyte and mast cell enzymes may play a si6nificanl role in the development of abwonn.lly high local concenuatiuns of an- Swweesin 11. associated with vsrious inRrnrnatory processes. Reilly, C. F.. Tewksbury, 0. A.. Schechter, N. M., asd Trovis, J. The Journd of aioloSkar CAnwlsry, 2S7(IS):l619-l622. 1912. OtArr sayr.rt: National lnriituln of He.l6. Fran the Department of Bioehemistry. U.ire+sity of GeaSia. Athens. d,e Marshfield Fouwdaliow for Modictl Research. M.rifidd. WI, and the DePartment of Medicine. [>vke University School of Medicine. Durlum. NC. THE EFTECT OF a; MACROGLOBULW ON THE INTERACiION OF a,.PROIYJNASE INHIBITOR Wfft1 PORCINE TRYPSIN The dissociatiow of a,-proseiut.e 1.hibNOrporeime tsypsin eomplea is rapid .nd h.a Persruned die esperirnents reporsod here. 1n this sludy the rsse o( dissociatiow of the a,-proseinase idub+orporawe a'ypaN earplea by itself wr comp.red with the rase of drsaociation of the same co.+plea is the presence of a; manoglobulin. Iw the p.e.ewce of the lanes inMbwar the du.ocitlioe was mote rapid. std active a; pro- leiease i.hiMtor could be rocoverod tw the tniaNUe. Howeva,.o s,ctive inhibilor could be detected .fser dts.ociatas tn the absence of a,-ssucroglobulin. This recovery of active a,- proternase inhobtor frrnn compkaes with porcine trypsia is the 6rst demon- stntion d a thcrmodynamtc eWdibr+um between this inhibitor and pdeinase. Coane- yuently, the lrtissfer of tzypun (rorn compkaes with a; Prokinue inAibilor to a; macroglobulin may be eaplatned.sd passive phenomenon which does nnt tequire a physical collision between a; m.croglobuli. sad dte a,-ptoleinase inhibitot:porcirte tryPsiI compie.. The dtssociwion of the coanplea occurs more rapidly in the presence of a; m.csoglobulin becawe this inhibitor compkses trypsia leaving the a,-psoteiwe inhibiwr.potcine trypsia oompka by both the irreversibk breakdown step and by rcversible diuaciation of dte tomPlea. Beatty. K., Travis. J. and Bei1h.1. alocAiw.kn et AiopAysko Acse 704:221-226. 1482. pttk.r s.irat: NMional Institutes of Ncallh. From the Department of Biochemistry. Univenity of Goorgia. Athens. and the FatwNW de Pharm.cie. Laborstoire d'Enzynsologie. UnivenN! Louis Pasteur. Strasbourg. France. ENZYMATIC REDIK'TION OF OXIDIZED a-1-PROTEINASE INHIBITOR RIS?(Nt!'S BIOL(XiICAI. ACTIVITY O.1datwN+ easily tnsctivates a,-proteinase inhibitor (a. PI). the major serum ie- hihoux u/ )+riacalylic aitivity The noted inactivation seems kr be due to the oauls- lion o(an essential mcthionine rcsiduc(s) in a,-PI that is required frx the inhibition of elaatase anivity. When niethionarc n:siduc(s) in a; PI we onidized, they result in MeqPl. la /ht:ory, the presence of.u esuyme in cells dw can reduce Met(U) residues i. pdeins 10 mnhionine provides a mechanism for seslainf bioh><ical activity 1o a praei. thM has been inactivated because of oaidation of an essential metbiowiwe n-sid.e. It has been shown elsewheee t!w eatraets of EtcArricAfo coli contain an entyme 1AM can toduoe Mel(0) residues is Ptokin a rtsedtioni.e. This study shows that oaidized, fuacctionally insnive c.nirc e; PI does in8xd repi. its biological inhibitory activity .fter reductioa with a paAi.lly Pwified prep.rnioF of E. cofl Mel(0) PePaide r~~ret.ae. Abrrro, W. R. Weinbornr, G.. Weisab.ch, L., Weissb.ch, H., and Brd, N. Prorrediwas of tAr NaNonof Academy of Scirwcrs of rAr United Statrs of Amwrrica 711(12):715 3-71116, 199 1. • tazAer a.or .rf: National lkart, t.reR, and Blood Institute. Raw the Department of Medicine, Albert Einstein Medical Ceetcr, Philaddphi.; Department of Biochemistry and Molecular Biology, Univenily of Florida. Gaiarcs- vilk; and Roche Institute of Molecular Biology, Nu1ky, NJ. I NEUTROPIIIL DEGRANULATION IN CADMIUM CHLORIDE-INDUCED ACUTE LUNG INFLAMMATION Gdmiwn chloride (CdCI,), a 1o1ic chemical 1ha1 has been reported /o cause centriWbular or scar ernphysem., was used iahis study b induce nnuroPlsil migrYien iMo the slveolr spaces as a model of hwtd infl.nunaios. Results showed Ihu lobtr i.tr.b.onchial instillation o(CdCI, (200 ptlml) is sd'ate c.usess teptoducibk.cwe pulmonary inflammation in dogs. The inAas of i./lanunalory nnutuphils from the circulation into the alveolar op.nes te.ches a eusMmua appoaimately 16 hars aAer the CdCI, aealmeM i. the tzeated hibe, while the eoMrdatenl lung appears twrmd. Morphmneuie qunwilatio. of peroaidue-posilive (azurophilic) gratwks in the in/lam- smlay aewsophib shows . 71% bss of these gwwles, wilh little or no loss of the peaoaidase-ne`nive (specific) Rrnwks. Thtse dafa ae in good .dseement with the wneasured loss of itNncellulr elsatase, r enzyme ktawn so be localized in the twrophilic gratwles. The results suggest dw debrawulation of .zurophilic granules twy occtr selectively dusing this chemically-induced scute i.Aanunatiow. Ysse.da, H., Damiano, V. V., Ts.ng, A-L., Merasze, D. R., GlasSow,1., Abnrro, W. R.,.ad Weinbaunr, G. Aiwrricon lourwof oJ ParAofotr 109:115-1 S6, 1912. 0/Aer snh..t: Nriond Heart, Lung, and Blood Institute. From the Research Division, Department of Medicine, Albert Einstein Modical Cen- ta, Philadelphi., aed the Franklin Reseaich ('enter, Phil.dclphia. PARASYMPATHETIC INNERVATION OF THE CERVICAL TRACIIEALIS MUSC'tJ? IN LIVING IX)GS U.e of Ihe trachea to stuJy airway (unctinn in livingdogs permiss es.rninNion of s prolaypic central airway under relatively cunvemenl circumstances using establishod ~ 12 1 33

tcchmdo6y In the study prescMed hert, lhe pathways by which parasympathetic hhers were .arned w the cervical lrachaelrs musck were .hxacten:ed in 34 aneslhetited dnRs Nerves were stimulated ekctrically, and tracheal tension was monaored in segments of the posterior membrane in riro. Stimulation of supcrior laryngeal nerves contracted 34 of t4 cranral cervical segments and two of four caudal ccrvical xRments. Recurnnl/arynseal nerves contributed b innervNion of 34 of 34 cranul, as well as (our of foui caudal, segments. Stimulation of Paracurrenl nerves contracted I I of 34 cranial aed fow of four caudal segments. Mechanical effects of eutpha`eal contrac- tan, induced by uimulatiwg p.rareerneM nerves, did not aher tension in tracheal segments. Tracheal contractions induoed by stinwlation of all lhree pathways simuka- atously were significantly less thr the awn of contractions produced by stimulMint each set individually; dris wn prabably dre lo anastomoses between ><rminal neurons, overlap in tlrcir.n.lomic diatribruion, a ielercellrlar nesuses in trac healitis musck. lt seems, therefore, dw parasyrnpadretk inwervation o( Ihe canine u ahea is by three drffereM neYroYUIOTK pathways. Brown, J. K., Shields. R. and Ciold. W. U. /orrnol oJApplied PAysiolory: Reapirtl. Environ. Esen:ise Physic,l. S3(3):617-625, 19112. OrM.r sspp.rt: National tleut, Lung and Blood Institute. Fmm the C.rdiovasculr Research Institute and Department of Medicine. University of Cdifornia. San Francisco EFf7:CT OF SMOKIN(3 A CIGARETTE ON THE DENSfTY DEPENDENCE OF MAXIMAL f:XPIRATORY FlAW Earlier studies of these investigators have shown Na1 tobacco smoke causes an increase in airways resistance and a drop in espirslory flow Y 10% of the vNal capacity (FEF.) In the present Nlernpt 1o gain further insight into Ihe nature and site of this brvrschnconstriclive effect, maximal espiratory flow was measuted in 12 heaNhy vol- wween while they wete breathing air and a bwdensily gas miature (heliwn-osy6en) bikxe nd after smnkinR a cieasdte. Resuhs showed Ma1 prior to smoking the forced vital capacity (FVC) measwed while breathing a'a was nol significantly different fsan that obtained while breathing the hdiumosygen misture. Smoking did aot cause awy changes in FVC. However, aher smoking. (he FEF. measured while breathing air docreastd si6nificrMly, while snwki.R did not cause any changes in the FEF.obtained a(tn breathing the low-density {as saistwe. AVnas. increased from 47.1! 11.4% Lefore smoking to 57.0 ± 13. 3% after smoking. There were no chantes in flow at 73% FVC srd volume of isoflow. These observrions re discussed in light of the equal pesswe point (EPP) analysis s,nd wave speed theory of flow limitation. It was con- chded dut after smokins. /low becarnes snae density dependent because there is consaictiow of a/low-timnin` segme~t downuream frum the EPP. located in lobar and segmental bmnchi. No acvle effect of tobacco smoke on the small airways could be dcnnnstraled. Tavein Da Silva, A. M and I/anwsA. P. R.rrirotron 41 258 262. 1982 Fnwn rhe flel+rtmcnts rd Medicine. Physiology and BNy+hysK s. (korgctown t)niver- say Salmrds ul McdNrne and Ikntlstry. Washm6lon, I)C. 34 I RISPIRATORY AND CARDIOVASCI/LAR EFFECTS OF INTRAVENTRICULAR CHOLECYST()tUNIN In this attempt to assess the rok of chnkcyslokinin- and 6astrin-like peplides in the central. regulatory control of respiralnry and ca.diovascwlar functions, chokcysb- kinin in doses of 1-300 ng was administered bNO dK 1Meta1 brain ventricle of chbra- luae-anestheliied cats while tracheal /irAuw, assterial blood pessure, and bean n1e were monitaued. Results showed thn the mosl striking effect was an increase in respiratory activity. This was observed with a dose as low as I nR rd peaked with a dose of 100 ng. Respiralory stimulation was indicated by an increase in respiratary minrte volume. an increase thM was due w an increase i.lid.l volume as aio sipificanl effect was nnled aw resprratory rac. MsqirMOrr tpnK, eapirawry lime, and total respi- nuwy cycle duration. On the other hand, when 300-1000 sK of chokcystokinin were administered intravenuusly, po respiratory stynWanl effecl was observed. These re- whs indicate 1hM chokcystatinin acts in 1ht brain to stiewlale respiration. Paaani, F. D., Taveira Da Silva, A. M.. /imnosA. P., Girvey, T. Q., III and Gillis. R. A. FwaPeanlownal oJPAo.wwcoloty 7d:129-132, 1982. OrAei s.rN..i: American Ikar1 Association. From the Departments of Ph.rmacology, Physiology .nd Medicine, Georgetown ud- venity Schools of Medicine and Dentislry, washington, DC. CIGARETTE SMOKE CONTAINS ANTICOAGULANTS AGAINST FIBRIN AGGREGATION AND FACTOR Xllla IN PLASMA In this in vitro study. Ihe effed of cigarette srnoke on fibriw aggregation wr inves(ipbd by the rse of «ater-sul.ble, tas-Phase eornpoweMS of smoke. obtained by bubblind the smnke produced fram one cigarette tlrorRh thsee ml of buffer or distilled wra. This estract was incorporated in varyirg dihMiorq in the buffer 1o which fibrie wrnomu solution was added in order to initiate fibrita aggregation. Results showod a do.edependenl delay in 6brin aggregation. hscseasi.R the amnwrt of smoke estncl teat.Ned in decreased abaorb.oce of drc clot and delayed onset of fibrin aggregation. The fibrin aggregation inhibilor was aho esaanwrcd by tue of two differing Abrin peparatiows wNh o chains lacking COOH-serseind segments. From the swn of these Mrdies, it was aeen (hat cigarette anwkt CorNairn two dislincy coaRultlfon inhibitors: one which prolongs the ckrttint times of plasma by inducirtR delayed fibrin aggregation and requirinR the C(X)It-lersninal teeRinn of fibrirt a chains 1o esert its effed: the other inacliva(es XIIla, thus preventing the cross-Iinki.Rof 6brin polymers. These anticoaj- nlant properties of smoke ase demonstrable in plasrru, where they may play a mk in the physiology of smokieg. , Galas.kis, D. K., Laurt.N, P., Janoff. A., and CIwnR. S. 1. Srirnct 217:642-645, 1982. OtAer sappe.t: l1. S. Public Ikahh Scrvice. From the Health Science Cenler- SIMe l)nivetsily of New York a1 Stony Brook. Stony Brook, and the Nuional Institute of Ihntal Research, Bethesda, MD. 33 1

PLATFtFTS INCREASE NEUTROPIIIL ADHERENCE IN VlTRO TO NYLON FIHI•R Inlera[tNM of neutrophils with platelets is a common phenornenon that may be impxtant in the patlwisencsts of various diseases. Although platelets are known to have a saun6 a/hmty for (oreign artaces, their effect on the measured adherence of neutrrrf+hils in Ihc commonly used nylon fiber systems had not boen detennined. Thercfoue, in the study reported here, the effect of platelets on the adherence of neulmphils to nylnn fiber was assessed in whole blood samples and purified neutniphil suspensions in the presence or absence of ptauna. Measurements showed that in whole blood sampfes, increasing nrn+bess otphftIda were associated with increasing adher- ence of neutrophds. Addition of ptMelets in plasma 1o purified neutrophil suspensions incrcascd (a<O 05) neutrophil adeseace from 76.2% 2 1.4 to 88.0% ± 2.0. Simi- larly, addr/ion u/ washed platelets wihoaM pl.seu also increased (y<0 05) neuu'ophil adhetencefrontA7.9% s 3.8 (widnrtaddedplalekts)to94.2% ± 1.6(withK10,000 plalekts/mm' added). In cowtr.st,.o wRmentrion of ttetttruphil adhereace occurred if platelets had their auregwiow tespowse suppressed by pretuewing platelet dnmws with aspirin. Scanning ekctron microscopy supponed these findings by showing plate- lets in close association with weutrophib adhering to sylon fiber. These findings emphasi:e the impanance of platelet wutnben and reactivity on tAe adherence of neuuslphils. Rasp, F L, Clawsoit, C. C and Repinr, l. E. TAr lorrnal oJtaDoratory and Clinical AleAicine 97(6):l112-lt19, 198' . Other ssqp.t: American Lung Association, Minnesota Medical Foundtdion, National Institutes of Ikalth, Amencan Ikart Associrion, Kroc Foundation, an1 the Graduate School of the University of Mtnnesau. Fsom the Departments of Medicine and Pediatrics. University of Miunesota Health Sciences Center, Minneapolis, and the WebdW.rin6l.ung Institute and Ikpartmeal of Internal Medicine (Pulmonary). University of Colorado Health Sciences Center, Deaver. A NOVEL MECIIANISM FOR PULMONARY OXYGEN TOXIC(1'Y: PIIAGOCYTE MEDIATED LUNG INJURY 0 In initial studies, a significant increase in the number of polrm«pMowckr kukocytes (PMN) in the alveolar lavages of rats esposed to hypctosia for three days was noted. 'flThis observation led to two hypntheses. The first hypothesis. thal hyperosia, causes injury to alveolar macrophases (AM), inducing their rekase of chernotactie (actnn which recruit PMNs 1o the IunR, was supported by evidence that:ll) hyperosia damages AM in rrvo and in ritro, and (2) ehemotactic factors from alveolar lavages ar hyfrrusra e sponed animals .re biochemically similr to those released by AM esposed w hy/+rn..u in.ell culture The second hypothesis, that PMNs play an important role ra pulrn.rry ua ygcn aurc Ny, was supptxted by the finding that the degree of endtMhe- hd darnagc ~as highly u.+clred wnh the numbcn of PMNs rccovcred in alveolar IavaRt% in twrh neuurq+hil sulhcrcnt and ncutnipcnK rabMls. However. two major unknowns slill remain. First, how ducs hypcroaia damage AM? Seaud, how do PMNs damage enduthcltum? At this time, a rtasonabk working hypothesis is that the damage in each case is due lo the fiwmalion of oay6en radicals, particularly the highly reactivc hydroayl radicals. Improved understanding of the nsk of AM. PMN and hydrosyl radical in lung damage due lo hyperosia will be important in elucidating basic mechanisms involved in the p.lhosenesis of pulmonary oaygen loxicity, as well as in other types of environmental lung injury. Foa, R. B., Sh.sby, D. M., Huada, R. N., and Repfae, J. E. CHEST ItOS: )S-1S, 1981. Other s.ypri: American Heart Association, National Institutes of tkdth, and the Mayta`-Crawford Trust Fund. From the E.perimenlal Medicine IXvision, Wcbb-Wri.g LrnR InstituNC, University of Colorado IkaNh Scie.ce. Ceater, Dewver: POTENTIAL MECHANISMS OF LUNG INJURY FROM HYDROXYL RADICAL Earlier biochemical studies h.ve indicated 60 itduled environmental tosias may, in one way or andher, contaibwe to hwt6 damage. For e.ample, there is evidence tha/ polytnaplwnwckr kukocyfes (PMNs) are efbcienl at making highly reactive OI intennediates a.d since facbra released (som alveolar m.croph.`es 1 AM) espoaed to hyf erwtia can stimulate n;leaae o/0, intersttediales (wm PMN, it appears likely that 0, iaernrodiatcs are involved in this injury. More seetaf work has foawsod oa the killing of bacteria by PMN as a model of PMN-indueed Wng damage. tn one of these Mudies. increasin` coacenlra8oes of DMSO or (hiowea progressively and signi- /kan(ly decreased killiaR o1Ssap/iylococnu mvtru, 302A, by normal PMN but did not forther decrease the abnormal bactericidal activity of COD PMN which fail so produce •OH. For swdies with Fe •• il was hypothesized Muf S. ar.eri was providing a m.jority of the subs(ances(s), rucA as Fe ••, which qrcn reacted with HO, to generate •OH and CH, from DMSO. The results of these studies idicale dw hydroayl radical (•OH) plays M impnrtant role in the killing of S. atr.nu by hunua PMN. Other sludies twMsted that •OH might also be farsned and participate in the killing of S. awe.u in anewher way. Elucidation of this IMter mechanism was an outgrowth ol sttdies dowe fo determine the bactericidal activity of chemical system, Fe• • and HO,. which ge.er- alea •OII. In these Nudies it appears that /1,0, rcacb with Fe" in die bat:teria b produce •O11 by a Fenton type reaction. Hydroayl radical then injures S. arrrrs by reacting with key organic molecules. II was also noted hese that DMSO prevents injury lo S. aurers by scavenging and detosifying •OH. I Repine. J. E. rt al. CHFST eOS:as-1aS, 19111. Other s.ypa.f: American Ikrt Association. National Institules of /kahh, ind the Maysa6•Crawfurd Trust Fund. From dse Webb Waring I.ung Institute and the Division of Pulmonary Medicine, University of CrrlrMado Health Sctences Center, Deaver. 37 36 I

~ IIYI)R(Xif:N P1:ROXInE KII.IS STAPHYI.(K'O(Y-(/S Af/RF.'US BY REACTING WIT11 STAPIIYI.(X'(X CAL IRON TO Ft)tM HYhROXYL RAI)1('AI. In the investrRatinn repxtcd here, an anemp was made to elucidate the mecha- aism underlyin6 the birtthemrcal role of iron in barlenal hosl defense intcrscliwms. Results shrwed that two different lines of invcslitalion suppxtcd the premisc 1ha1 k111M6 of Sraphyfo(-At'ru! OWful, 502A, by hydrogen pcroside involves (onnalion of drc tnnre tosrc hydrosyl radical (•OH) 11uoVh the inm-dependent Fenton reaction. First, growing S aMrrrs ovaeiRlM in brolh media with increasing cnrKenlralions of imn incrtased their cnntenl of uow and drartically enhanced their subscyuent suseep- tihrhty to killing by H,U,. Sooowd, in direct relation to their effectiveness as •011 scaven6ers. thianea, dimnhyl thiourea. aod'wm bentoate. and dimNhyl sulfoaide inhrbeed H,p; me<h.1od killing of S. aarrus. Therefore. it seems from this work dW, while inw is an essential `mwth nutrieal, il may also povide an "Achilks heel" fex S. awrr.+, indicating Ihat the reporlod wrochanism may be a new way in which iron is beneficial to the host. Rrpinr, l. E. Foa, R. B and Berta, E. M. TAr lournol oJAiolo`icof CAnwlsrry 2S6(14):7094-7096, 1931. Ot>.rr s.#/.rt: National Institutes of Health, Americaa Heart Association, and tbe K.oc Foundrion From the Webb Waring l.ung Instituae and the Pulmonary Divisions of the DePan- rnents of Medicine and Pedratrics. U.ivenity of Colorado Heahh Sciences Center. Denva. SERUM FROM PATIENTS WITH INVASIVE FUNGAL INFI:CTLINS INIIIBITS TIIE ADHI:RtNCE OF POLYMORPIN)NUCLfJ1R I.LrJK(X'YTES AND ALVEOLAR MACROPHAUES Ahhough considerabk aneMion has been directed recently tow.rd detennining dre effect of bacterial infection on neutrophil adherence. little anentica has bee. paid so far to the influence of fungal iskctioo ou phaSocyu drcreece. Ia the iavesti6Mion repoAOd here, howevcr, the adherrcnce o(vrious cornbinatioas of po1 /murphonockr kukocyw (PMN) or alveolar eucrophqes tAM) and serom from pN.ents with fungal infections or from corMrol subjects was evaluated in virro using the standard nyloa bber pipette technique. Results showed that the intrinsic adher<nce of AM and PMN front Pricnts with a wide variety of urweahd fungal infections was rKxm.l, but 1ha1 these patients had a serum inhibitor that could tevenibly decrease 1he.dhen:nce of PMN and AM Specifically, studies using various combinations of PMN and serums from pa- /ics>ts with blaslomycosis suuested thal the adherence defect was due to a serwn disonkr rather thal an intrinsic cellular abnormality. While peincubation in sesvm from patients with blas/nmycosis decreased the adherence of control PMN, pdncuba- tion in cnntnd sen+m axrected the decreased adtineece of PMN from PNieMs with blasawnycnsis On the hasis of these and other related studies, il was concluded that the intno.K aJhrrcnce of PMN and AM from patients with unlrealed fungal infeclions is n46M.l. but that rhr.e patients have an e.vinsre heal-labik serurn fachu Ih.l can dnreasr the nlhrrrn.e rPf I'MN and AM 1a iL. Rasp. F. L.. tiuosi, G. A. and Rrpinr, J. E. Anwricnn Review of Rnpirurory Uisrwr 12):6M-6J9, 1961. OrArr suyorf: Minnesota Medical Foundalirm, Minnesota and American Lung Asso- ciatiwn, National Instnules of IleaMh, Anrrica. Heart Associatio., and the Kroc FirundNiun. Fiom the Deparfinents of Medicine of Ihe University of Minnesda Iledth Sciences Center and the Minneapolis Veterans Administration Medical Ce.1er, and the WeEb- Warin61.un6 Institute and the Department of Medicine of the Uaioenit)r of Colorado IkaNh Sciences Center, Denver. ANQIOTENSIN C()N VERTIN(3 ENZYME CONCENTRATKNVS IN THE LUNO LAVA(;fi (W N()(MAI. RABBITS AND RABBITS TREATED WtTll NITR(X;EN MUSTARD EXPOSED TO HYPEROXIA Granulocytes have been implicated recently in the deve{opment of acute Ay- perosic lunt injury, an edematous IunR eishaP IhM is characleriytd pvssly by massive edema and histolosically by e.dotlu;li.l iujury widt petivsculr in8amrraioe. 1N tk Presea sthdy, specifically, inenea.ed cawcerwaliwu of a.tiolewsift eowveni.g ewryme (ACE) were found is WnR Iav.Res (eara rabbits eaposed Wr 72 hours ro hypnoaia and the concentrations of ACE were correlred wi1A rasioa of eatravascular hug walcr ro body weighs and albrwnis coucenvyiwx in Irrng Iw.ge.. In parallel taudies. rabbits eealed with .itrogew mustard is which tr.wulocylopeeia was maintained duaylrou/ 1Ae 12-how hyperoaic eaposute period had kss eiidegiee of edemnous hsngiojury a.d bwcr cwrcearatiows of ACE is their haK lasqes dwt saailrly trered rabbiws i. wlach Rrarwlocylopeaia waa aw1 maittaiwed. 7Uese resrNs suuesled thal Rranulocytn eowtribu/e to acwe edematous lunt i.juryy from wypnoaia aad th.1 ACE eorrewalian in hutR lava6es reflect dris process. Shasby, D. M., Shasby, S. S., Bowrna., C. M., Foa, R. B., Harada, R. M., Tre, R. M., and Rrpiwe, J. E. Aiwrriren RrvJrw of Rrapinorwy DJuae 12d:202-202, 1981. OrAer r.yr.rr: National HeaM. Lung and Blood 4wkwe, American Heart Asaocia- tiaw, Naliooa11es1itwes of HeaMh. aed Ihe Kroc Fowdrion. Fnvn the Webb-Waring Lung ItwipMe, DePattmerM of Medicine IPulmo.ary Sd- eneea), l;nivenity of Colorado Health Sciences Cener. Deaver. NEUTROPIIIIS AND LUNG EDEMA: STATE OP THE ART This brie(rcview focuses oa Ihe emer6ingrule of eewrophils and their producu is the development of aduh respralory disUess syndrome IARDS), the mou common prescntioR form of edematous hmR injury. For some time, the association of ARDS with multiple inciting events has led 1o the presumption 1htl, tander certain circum- srances, many factors might be involved in the palhogeaesis of ARDS. While so1 /bwdy established, a M.sa nf alvealar-capllary, mmembrane integrity which resuhs is lung edema appean to be a pathulogic change conunon to all cases of ARDS, Mus

makin6 it prssrbk frK ctllulu or humoral components to contribute to endothelial injwy. A number of recent observanions suggest that neutrophils cuntnbutc to the devewoprnent of edematous luns injury. Firsl, increased numbers of ncwnh+hils are c,xrumrnly found in lunt lavages of patients with ARDS and early in their illness. Second, nculn>rhil depktion pnNecu animals from esperimemal edematous lung injury, and third. ncutmphils have been shown 1o have polenl mechanisms for causing tissue injury, aherins vascular permeability and petturbint hemrdynamics. Overall, cnrniderabk evidence is now available 1o suggest that neuttophils participale in the development of acwe edematous lung injrry such as 1ha1 seen in patients with AR()S. The mechanisms by which neutrophils ntighl medine these effects appear to be many, vrraf and still ill dcfined M this time. R.oinr. J. F... Bowman, C. M.. and Ttle, R. M. CIIf.'ST OIS I7S SOS, 1" 2. O4.r snrywf: NMiond InstNwes of IleahA, AmrYican Ilean Associaion and the Knrc Foundauon. Fmm the Webb W aring l.ung Institwe. University of Calotado Health Sciences Cen- tcr. Denver ALVEO(.AR MACROPIiAGE SECRETIONS: INfIIATORS OF 1NFl-AMMATNJN IN PUl-M(NdARY OXYGEN TOXICITY? In the paper presented here. Ntempts were made 1o elucidate mechanisms respon- sibk for ncutrophil rnflus into the lunp srd 1o uaderstand more clearly the contribution ol.eutrophils so the undefined pathogenesis of pulmonary osygen losicity. For this pudy, dvealar macmphaecs (AM). (olbwinglavage ftom lungs of infeclwn-fnee New Zealand White rsbbrts, were esposed in culture 1o normoaia (15% 0,) or hyperosia (93A O,) far periods up to 72 hn. Supetwstanls fran AM cultures were evahwted for yKir chcnuMactic, adherence ainwlating, and superoxide radical stimuWint activities for newnSdwls. Results drowed 1hN hypnosia damages and stimulates dveolar mac- sophqes /o release fatlors which affec( aertrophil recruitment, adherence and activa- tion. Preliminary characleriratioK of these (acton suggests thn the facton have aeparale identwies. Factors derived itom AM eaposed to hyperosia differ in moleculr weigM, hcr stability, .nd time of maainal activity. The stimulus for macro~ release of these facNxs may invdve dans.Se to AM by hyperosia. These observations support the possibility du1 AM and the proposed mechanism of nearoph.l recruitmeet and activation may be imponanl in Ihe patAoReaesis of osygen Io.icity and other (orms of aeule hsng injury. Handa, R N., Bowman, C. M., Fos, R. B.. ard Repinr. J. E. CHF.ST S1S:S2S•StS, 19s2. Other sart.rt: American Lung Association of Coloradn, American Ilcan Associa- ti4m, Naliireal Institutes of Ikdth, the Kroc Foundation. Hill Foundalion, Swan Foun- daravn. and KkberR Foundation I rorn the WcMA Wanng I unR Insrnure. Pulm.nary lhvnNwts. l/nivenily of ('.oldKado /Italth tit wna. t tnrrr Ikn•tr I OXYGEN RAI)ICAI: INDI>CED PULMONARY EDEMA: A MI:('l1ANISM FOR TIIE PR()0N1Cf1(Nd OF NONCARDI(X;ENIC PULMONARY EDEMA BY NEUTROPHIIS Since stimulated ncwruphi4 auke O, radicals snd since 0, radicals h.ve been shown to damaRe a number of biological tissues, it seemcd possible thal release of 0, radicals from stimulated neutrophils might pertwb the dveolar<apllary membrane and lead so proteia-rich edema fornwion. To teal this hypolhesis, isolated usod lun6s from New 7.ealand White rabbits were nwnitored siid, after a stabk baseline periad, purine and samhine osidase were introduced into the pextusMe with or without prior introduction of O, radical acavenRen. Results of this investigation showed dut 1he chemical generation of O, radicals by intravascular knjoclioa of pwine and santhine osidase resulted in acuue pro/ein-rich edema formation in the isolated Wnp. This edemsiaus process was nurtedly inhibiled by the prior iadividual injeclion of several O, radocd uavensers, including catalase. dimethyl sul(oside, and dimethylthiarea. These findings provide direct evidence lo suggest dnt O, radicals are capable of perturbint the a'a•blood b.rsier.ud cwsint a prolein-rich edema. They also suggest thr 1he intrs-vascular release of 0, radicals from /Iinwlaled neuUVphils might be capable of producing a simil.r acuwe edemalous hrillf iyury in certain clinical seuinp. Tde, R. M.. Shasby, O. M., Va.Bentlwysen, K. M., McMurthy. 1. F., and Repine. J. E. CHEST tIS:37S-59S, 19112. ON.r r.yprt: American LrrtR Associalion of Colorado, Atnerican Ikart Associa- tion, National lnsti(wes of Hcahh. Ue Kroc Forndrio., Hill Foundation, Swan Fou.- drion, and the Kkberg Fowdation. From Ihe Webb-Waring Lwrg Iwilule. Pulmo..ry Division, Cardiovascul.r Pularo- .ary Research l.aboratory aed the Departoers of Medicine and Pbdiatrics. Univenity of Colorado Ikalth Sciences CesMn, Denver. RFDl1CTlON OF TIIE EDEMA OF ACUTE HYPEROXIC LUNG INJURY BY GRANUTACYTE DEPLETION 11e rdationship be(ween grawloeyta in the hsnR awd tYe edenu of acwe by- perosic lu" injury was eaansinod in the study reportod hese. Results showed dW alUrough increased numbers of Rranulocylcs are found i. /wrgs acutely injaed by hypesoaia, their contribution to lung injury remains untsow.. h was fn.nd thr cinrt- IatinL grarulocytes increased mrtedly israbbits e.posed so hyperosis for 72 Ys. rd Ihr tJre number of grannlocyles in Iwsg lavases also i.ceeased and wpe eomelued wit\ the de6ree of edemalaus lung injwy. FuAAersnore, when nbbNs were 1realod widt niuoten mustard (1.75 rng/tg) and devdoped sustained grarwlocytopenia, eaposwre to hypertosia for 72 hn. resulted in fewer grrwlocytes it IuaR lavqes and less edema- bua Iung injury. In cuntrast, when rabbits were similarly treated with niuogen mustard but did not maintain sustained grwruMrcytoQenia throughout the esposure 1o hyperosia, iecstaaed numbers of tranulocytes were found in lung lavalles and the degree of 441 1 41

edematows lung injury increased to kvcls na differem (nwn those observed in nsygen- espand rahMts thal had not been treNed with nitrogen mustard These tindin ss suggest that granulocytes may cnntnbute to production of edema in acute osy6en 1isicny. Shasby. 1) M.. Fos. R B, Harada, R. N. and Repint, l. E. lrwrnof aJ ApplieJ Physioloay: Raspira. Errviron. Ezercitc PAysiol. 52 (5): 1237- 1244. 19e2 ThAer srh.rf: National Ikrt, Lung a.d Blood Institute. American 1leart Associa- bon. and the Kroc Foundation. From the Webb-W uinR 1-ung Institule, Department of Internal Medicine (PuImonary Sciences 1)ivisan). University of Colaado HeaNh Sciences Center. Denver. GRANUId)CYTFS MEDIATE ACUTE EDF-MATOUS LUNG INJURY IN RABBITS ANI) IN ISOLATED RABBIT LUNG PERFUSED WITH PHORBOL MYRISTATE ACETATE: ROLE OF OXYGEN RADICALS Many forms of acute noncardiornic hmg edema are associated with an accumu- (ation of inRanrnawry cells in the alveoli and microveasels of the hay. and there have been many suttestions that the activated grarwlocytes play an importanl role in this pncess. In the sludy reported here esplicNly, it was seen that intravenously injected phutbnl myrisate acetate IPMA) caused a polcin-rich edema in lun6s o( control rabbits but aot in granulncytopenic nbbtts petre.led with nitroten mustard. Speci- Ikallyb eontrol rahMts treated with PMA had higher lung weigIM to body wei6hl ratios and hrnR lava`e albumin concentrations than `ranulocylopcnic nbbiU pctrealed with nitrogen nnnWd and then given PMA. To further clarity the rok of granukrcytes in the poduction of edema, additional esperuneals were conducted in an isolated p:rfused rabbit hatt. Additinn of prified `rsnwlocytes and PMA to the balanced salt p:rtusak caused king edewra. whereas neither pnwbcytes na PM A alone caused edema. Also, in order so determine dte contribution of oaygen radicals to the prhoeenesis of the edema. chro.ic grrwlomrota disease granulocyles. which are deficient in osy6en radical p.odretionc were added to dte isolNCd lung p:rfusfte. Chronic Rranulomataus diseaac Bratw{oeytes.nA PMA did s+ot cause edenta in isolated hNtp, whercas srawlo- cytes (tvan normal hunun subjects and PMA did. These data suggest that osyten rwbcals tek.aed Inwts slimul.led tn.ulocylt:s contribute to the pathogenesis of acute edematous lutt6 injury. Shasby, D. M.. VanBenthuyses, K. M., Tsk, R. M., Shasby. S. S.. McMurtry. L. • and Repine. J. E. Amr.icen Review oJRespiraory Dissate 123:441-447, 19i2. (k4r sappari: American IkaA Association. National Institutes of Ikadh, and the Kroc Fuundatiott From the Webb Waring Lun6 Institute and the Cardiovascular Pulmanuy Research IJfx"tory of the Univenny of Colorado Ikahh Sciences Center. 1)cnrer, and the 1'ulnrmary IHvisum o( Ihc Univenity of Virginia School o( Medicine. CharkNlesville. r CYT(K7IALASIN 8 AND THE STRUCTURE OF ACTIN ci1:1.S 11. FUR7111:R EVIIH:NCE FOR TIIF SPLITTING OF F-ACTIN BY CYT(AYIALASIN B Cyuxhalasin 8, a fungal metabdile that can alles cell shapes and inhibit a wide variety of cellular movements under certain cond'qions, has been shown in ahe pasllo be abk to reduce the nctwoMic struclure o( actin filaments. 10 this cotrwrxmicrion, additional evidence is presented that cylochalasin B shortens aclin filaments and that this shortening takes place without aet dcpnlymerization. Overall, the work done here showed that cytochalasin 8 decreased the Bow birefringence and S., and increased the estinctinn angle of actin 6lamencs in salt sohuinws favoring polytneriztlioe of the prneia. These changes occurred without a deKclabk increase in the equilibriuns actin trrn~omer concentration detertained by a radioassay. 71rcse results eomplaeeM eartia observations indicating that eytochalasit B shortetn actin filaments without .et de- pulymerizatirwt. Analyzed in lenns of Flrry's classical network lheory. this shortening accounts row the marked effect of cywchdasia B jtt dissoFving the Sel stnrcture of F- actin ennslinked by actin-biwding protein corrcentratiow for incipieM gelnion. Cy1o- clulasin B decreased the annealing rate o( low concentrations of anin Bl.merw fragments pepaned by sonic disruption. This result is consistent with the idea that cysochalasin 8 binds lo the ends of aclin filaments, and may esplain how cytochalasi. B causes filament shortening. Maruyama, K.. H.rtwi`,1. H. and Stossel, T. P. BiocAimico e/ BiopAysiru Ano 626:494-500, 1990. OrAti saw.rt: U. S. Public Health Service. From the Department of Muscle Research. Bos1o. Biomedical Research Institute' Iknutub6y-Oncolo6y Unit. Massachusetts General Hozpital. and the Department of Madicine, Harvard Medical School, Bos1o.. STRUCTURE ()F MACROPHAGE ACTIN-BINDING PROTEIN MOLECULES IN SOLUTION ANl) INTERACTING WITH ACTIN FILAMENTS Evidence is presented is this papet thal anis-bi.ding ptotein is a dimer, which has the capacity b initiate and popa`tMe isotropic adiw Nlaroea networks. For this partic- tdar study. 1Ae aructure of anin-bind'wtR tttulecuks wn esamiaed in solution and iMeracling with actin filaments. At physiological ionk stnen6th. acsin-bind'atR protein hr a M, vahre of S40 x 10' as determined by diroct and indiroct hydrvdynanwe memuremcnts. It is an assymcttical d'aner composed of 270 x 10' daltow subunits. Viewed in the eketrott microscope af/er negative staining nr low angk shadowing. actin binJin` protein molecules assumewbroad range of con(orseMions varying (rota closed circular structures to fully eslended strands. Aq eonliRurations are appnently derived fran the same structure which consists of two monomer chains connected end- lottd. Other observations noted in this papes indicale that aclin-bindin6 protein dinmrn ae estrcmely Ileaibk. In further studies. direct visualization of anin binding pnNein molecules between actin filaments in the electton microscope showcd that dimers are sufficient for cnrssbrid6inr of actin hlamenls and that acun binding prnein dinsen ne bipolar, composed of rrKwMUnen caarccted head to-head and having actin- 4) 42

A.. binding snes UK•alcd on the /ree lails. Overall. N seems a(K+arcn1 fnom these cludics that aclin binding prt,tcin is a dimer at physw1klglcal iwmic strcn6lh; earh dimer has two n1in hlament binding snes, and is. /here(rxe, wf/icicnt lo Rc1 aclm hlamcrNs in s,dutw.n. The length and Acxibihly of the a.tin-bindinR pwein su(wnus render Ihis molecuk sMUCiurdly suiicd for the crosslinking o( Iarse helical filam •uts on1o isrNnipic networks Hartwo6, 1. II and Srouel, T. P. Jowwol oJHoln rlar Sido4y 141:563-3i1, 1951. (Nier ssw.rt: U. S. PubIK He.k! Servke. From the Dcpanmcnl of Medicine, Mssaclwsctts Cxnerd Ilospitat, Boston. t)ISTRIBUTN)N OF ACT1N-BINDING PROTEIN ANI) MYOSIN IN Pf)I.YM()RPIKNVUCI.EAR LEUKOCYTES DURING LOCOMOTION AND Pf1A(:(K'YT()SIS There is a Iheory ,hat aclia and ptaeins that associate with is are the major ekmenb of bconsaw+e and Obatocylosis by Polymrxphawckr (f/NN) kukocyles: and tAn theory is we0 supfwned by the malernal presented hete. 1. this study. indinDct inunwr,Aoun:scence was used 1o esamurc the redistribulion of myosin and acli.- Mndin` prvteoa (ABP) mokcuks in rabbit PMN kuhocyses during Incomcwinn and phaRocylaus In unpdanted PMN kukocyln, ABP rxd myosin had a diffuse distri- hutKIn with some PredikatNle ftr the cones. In pdariied PMN kukacyles erawhn` toward yeast panocks, myos.n and ASP sluning concentrated in the anterior peuda ryrd In PMN kultacytes fiaed during phaRocyloiis of the yeast partrcks, antimyosin and ami-ABP staining concenttated stnhmgly in the distal portions of the pseudnpod embracinR the yeasis SuininR for caulase, a cyloplasmic paein in PMN kukocyles, for Iactoferma, a potcin of specific Srssw{es. and for myeloperoaidase, a pro/ein of a:yrvrhilic 6ranuks, was not eoacentralod in pertdopods. Taken Ngether with avail- abie rmxplw+b6rc and biochemical infornulion. these findings arc consistent with a mechanism wherein inleracliaws of aNia. ASP and myosin redistribute conicd eyto- plasm into peuduPods involved in beaaolioa and plyRocytosis. Vakrirs, N. {f., Stesdahl, 0.. HartwiR, J. H. and Stouel. T. P. Cell 24:191-202, 19e1. From the Department of Medicine. Harvnd Medical School. and the IkmNoloRy- Uacoh+Ry UnN. Massachusetts General Hospital. Boston. 11>F.NTIFICATION OF GELSOLIN, A CA"-DEPENDENT RF.GULATORY PRnT1:IN OF ACi1N GEI: S()(. TRANSFf)RMATION, AND ITS IN11tACF1LUUIR DISTRIBUTION IN A VARIETY OF CEI1S AND TISSI II•S (:clrlliw, s 91010 dadnm globular prolcin fnwn rabbn lung macrnPha6es, has Mes strnra hcf.rr to hc a marw ('a"-dcpcndenl regulatory pnMCm of aclin `el-sol I Iransfrsrmatwm. In the work reported hcre. antiserum prepared against Relsolin was rsed to detect the presence of poMeins immunologically relaled to 6clsolm in a variety of cells and lissucs. Results shawed that a single band of cross-reactive malerial which comi6raled with macruplu6e adsoliu was foud in at least nine different kinds of cells and tissues derived frorn rabbits and humans asd in four lines of cuNwod cells from humans and rals. Gelsolin was also identified in btwnaa srnsm and plasma, raising the possibility that it nuy conlribwe 60 the ckarance of actin from 1he cwculMOry syslem. Relalesl studies, using iedirecl imnwnollowescenl staining of acelone-fiaed m.cso- Phaaes and pdyrmxphoasckar kukocytes. showed tlw Relsdit tesides iw thp conical cytoplasm and that during phasocylosis it is eoncesMraled in pserrAopodia engulfing prticks to be ingesleA, an area of 1he cyt<Vlasm aclivefy engaged in awvemcnl. 1s k>ngiludi.al cryustal stxtions o(coatracKd rabbil skeletal sswsck, aMiRelsolin staining was associated widr Ihe I-baaid of tbe nsyofibnl. suggesting 11W it may be involved. by r% as yet wwkfined rnochawism, in skektal awsck function. 1o conclusion. these fbdinp ne compatible widt Me idea IIuH Reboliw is aw iaiteRrd prt of Ihe motile apil.ratw of ph.6ocytic cells and thr it reRalses ceu movement by changing the consistency of the cytoplasm. Yi.. ll. A.. AlbtcM,1. H. and Faluwm. A. (Stossel, T. P.) Tlk Jownd of Crl/ Qiology 91:901-906, 19l1. , t7rArr a.rh.rt: U. S. Public Hedlb Service and 1he Edwin S. Webster Fou.drio.. Fraa Ihe lkmuoloRy-(h,coloRy Unil. Massadwsetts Oeneral Ilospiul, and the De- prtmesu of Medicine. Harvard Medical School. Bostuw. ACTIN FILAMENTS AND SECRET/ONS: THE MACROPHAGE MODEL la dds fan-61kd book chapter, the RlMionship belween aclM filame.ts a.d secretion is scrvlinized in several difkaal ways. to one c*aper sadios, mapbologi- cal and pharmacological evidence is preseMed for dte participMion of actia snicsofila- meats in secrelion: wbik. in another seniow, a manophqe model is employed to cb.racterste the demonstrable association of tnicroftlarrM KartanRemeM aad secR- tion, it is noted heoe thr, i( Ihe eortical actit microf+l.nKM Inlice of nucropbates is to have some active or passive function in secletioa. it nwsl be capable of direcliaw.l seovemenl. The ekments of directiorul movement ase (1) a force-Re.erasisll mech.- aism. (2) an orienting inRueace on the force to provide direc/iuna/ilyc and ()) acontrod mechanism eserted on the forte-6eneratinR system or on directiondity or on both. Each one of these diredional eroveme4M elements in pesealed in fuller detail here. la sunrn.ry. the force-RenentinR mechanism is a wperpeeipitation of actin and myosia 61.menta. a process requiring hydrolysis of ATP and presumably based o.lhe slidiwg- 61.mesit inuraetion chader.zed in striMed snwck. llwse.essy-depewdenl saocbuiiswt may be a nujor caisumer of the macroph.Rda metabolic activity and can acco" for Ilse susceptibility of secrc/ion to inhibition by metabolje poisons. Direcliondity a.d awipldicMiua of the (orce generMod a.ise for eonudkd foed changes in the csvsaliak- ingot actia filaments. Gelsulm, a cakiwn-activabd pvtei., controls lattice ri#idNy of 44 1 45

i i actin by scverin6 .clin filaments between puinls of cn»slinkinR by actin binding prutcin lhe frce cakwm c,xrcdratN,ns that regulate thc activity of this panctn are kvels found in hving cells This mechanism for directiunal cyt.K+lasmic nrwcnrcnt camrol by cakwm is called the "Tugof-War" hypolhcsis. and evidcncc that thts meahanism earsts in macrq+ha6es is presented in this paper. Srnrsrf. T. P Mrrfbds in Ceff fliofogy 2):213-229. 199 1. OrAer sryp.rf: U.S. Public Ikahh Serrice, the Edwin S. Webster Fouedatiun and Edwiw W Hiarn. Fmm the IkmatoleKy-Oncololy Uni1. Mauaciwsens Genen111osQital, and the De- pusment r.f Medicine. Harvard Medicd Sdhod. Boston. TIIF. MOTOR OF Pt1AGO(:YTIC LEUKOCYTES There we three things that are noedsd for eytoplasmic movemew (1) furte 6encTtuon. (2) orientation of the faree 1o provide direction and (3) a copNrd mecha- nism. The present review sununari:es infonmationconcetn'rng the biochcmicalcampo- nenls of the mrnor of tnammalun phagocyus and how they can intersc. to generate dirmirmal movement In the first place, the nrolar region of pha`ocytic kukocytes af+pean 1o reside in the periphernl cytaplasmbeneath the plasma membrane. Transmis- sion electron microsraphs of thin sections of bxed ph"ocytic kukocytes reveal that this cortical cytoplasm consists primarily of filamentous tsulerid, and thMAbe 61a- mcnts have the dimensions of actin polyrners. These actia 61.ments seem to form a meshwork iw which the filaments intersect or overlap at random intervals. Myosin nrdocuks (abow 196 of the total proteis) of phagocytic kukocyles can furm bipular blaments at physiulosic plt and ionic suength, and these myosin 6lamente. bind to actiu filaments. Although all of the evidence is arot it yet. M this tin+e it seems possible dat: (1) the macsvphage cortea is an isotropic lattice of anin filaments cross-linked by a pauein called actin-bindinR pdei.. (2) Myosin 6laments, dispersed together with mageesium and ATP throughout the lanice, esert tetssion on the lattice by means of the ctossbridge nsechanism, and ()) dependi" on the cakium cancentntion. gelsdin. a receuly ditKoverad nucrophage poteia. tsplates the structure of the lattice. Corrmp.r- isoaa we rnade in this paper between the nwtar of manunalian phaaacytes and the maor of muscle fibers. and consideration is given to the possible im(rotunce of this rtrwor mechanism in cancer biology. Sroasrl. T. P., HrtwiR, 1. H. aad Yin. H. L. 1.: Saunders. Daniels, Semw. Rosenkld, and Denny (eds,): Fundontrntaf Mrt Ao- ruseu in Hriwon C'anctr lnaurwlosy. New York: Elsevier North Holland. lnc., 1" 1, pp 259-27). (AA.r aw..{: ( I S Public Health Serv/ce, the Edwin S. Webster Foundation. Edwin W llssnt. a.d (' 1. awd 1 1) Kwfnuut Frs.m the Hew,r.d.yty t1n...MKy t)nit. Massaehusens General IlospNal, and the ()e- I+.rsmrM of Medn inc. IIaL.Yd Malkal Ssha>t. Boston. 46 I 1)ETI:RMINANTS OF FORCED EXPIRATORY FLOWS IN NEWBORN INFANTS This study was undertaken to delineate the ntechanical propcrties of Ihe hnri in early life by eaamining the detertniaants of forced eapirMory flows and tbe respo.se of /lnws to I(eO, in newborw boys and girls. Spocifically, maairnd flows at functional residual capacity (Vmas,,, ) frorn partial espiralory Aow-volwne (PEFV) curves (achieved with rapid compression o(the chest) were obtaimod oa 11 heahhy tsewborm babies. MeanVmax,,,, size a'arrecled by dividing absolute values by raeasured thoracic Sas volume (TGV ), was 1.90TGVds. Specific upstream co.dretrces were hiRh. aaA tlr cmu-sect'wnal aea of the fbw-WoitinR sepnent was eslirwdod a be apposi- nwely 0.)0 cm' iw the throe infarw on whom recoil pressures at FRC we.e abo measured. The cross-sectianal area of tAe teajor bronchi in the weo.re is approsi- mrely 0.2t10.30 cm'. PEFV curves were coavea to the vohn.rc aais. Many of the nonrrales wtcrcasod drcir Aows while brerhioR a heliwsotyRen tas wtitture. These results suggest (1) sise-conected Ouws we higher in the t.oowtle Ilnt i. older chil6en or duNs; (2) the site of the Ilow-laniting segment r FRC during wtaaiesal e.pirsfay maneuvers is in large proaireal afr.rays, awirrwlar to the atlrll; and (l) the relMio.ship a/ airway sirs to parenchymd aire auy, be sirnilr i aeowares and adults «, i. 6ct, lirways may, be larger, relative to parenchym., i..eo.aes. Tbnefae, tAese physio- logical dau do ta1 suppxt the hypodtesis. based ow pnholoRical sludies, thr periph- er.l airways are disproportionately st.nller (when corttpned with cealral airways) is i.fasw tlum in aduhs. Tarra;g, L. M. er at. Jorrwd oJ Applied Pi1ya(olosy: Respiret. Env/ron. Esenchra PAysiol. S](3):122U- 122 ), 1982. O&tr nffoat: Natioud I.atitwes of HeaLlf. Frorn the DeRartmnr of Pediatrics. H.dssah University Hospiul, Mt. Scopns, ksr- salew. Israel. CAPTOPRIL: ASSOCIATION WITH FETAL DEATH AND PULMONARY VASCULAR CHANGES IN THe RABBIT (1") Captapril (D-7-snercapa-2-nsethylprvp..oyl-L-proline/ is a ndter new, onNy effective ithibitar of .nliatensiu co.vesling eozyme, which As recently baew s¢ poved for the treatmewt of refractory systemic hypertensio.. Adtssit.iauation of this apnt has been shown espearnerttally to decrease circulating levels of anRiolesui. U and to incrrase levels of br.dykaua and prostaglndi.s. For the Nudy pesnwed here, the effect of oral captopril ow ktal survival was assessed in pegeaa adrlt New ?talnwl WhNe rahbits (20 treated with dpopril and 12 controls). Resulu showed tf.r ktal death in the treated rabbits was 86%, in contrasl to IS in eotwd saEbits. Sane of the rabhits were nude hypo:ic in a hypobuic chsanber (322 wrn Ht peswre)dirirtg the period of captopril administration. Uwder these eowditio.s. captopril.drninislerod to the materwal rabbns tod a demunstrabk cardropulnron.ry effect in dte neoaaes, ay ahowa by a si6nifu: ant reduction in pulmonary arteriolr medial thickness and both left aad right ventriculr weights compred to the hypoxic wNrealed eoeuds. In view of these ubservatans. it inould be prudent to avoid using captopril Wr the treatmea of 47

hyprssensinn durinR prrRnancy, until the mechanism of fetal death and the reaxw , IIM %pCl'KS varlalMnl arT klNfwn. KeiM,1 M, Wilf, ! A, and Weir, E. K. Prrx«Jin`s of the Su.'reryfw EsPerirnanta( eioloay onJ MeJicine 170(11:)78-J8I, 1982 NatMwsal InstAules of Health. (MArr suhert: From the Ikpartnsenb of Velerinsry Science, College of Agricultural and Lde Sciences and Aneuhesiolo8y, Medical School, University of Wisennsia. Madlsnrs. and the Veter.ns Adrssisdstntio. Medical Center, llnivenity of Minnesota, Minneapolis. STANI)ARI)IlAT1ON OF N()RMAIDEIIYDE•INIx)CED FI.lNNt1:S('hN('E ANI)11S MEASUREMENT TO Ql1ANTIFY SER(7T(kV1N EMISSIUN IN PIII M( )NARY Nl:UR()ENfX)CRINE CE11S The fnrmaldehyde-induced fiuorescence (FIF) method is widely used for visuaf- icstwm and Quarwitnive analysis of bio8enic amines in lissues. In the paper presented here, a modd+ed and slandardized qurMiWive FIF procedure fnr producing fluorophores and ineasurint emission intensity of seraonintontainin8 neuroepithelial bndies (NEBs) in the rabbit lung is described. This technique, using epifiuorescence, was reproduced without significant differences between control groups. Important consderatinns for repmducibdily were: using the same humidity (80% RII) and reac• tion time (2 hn. ) dunn8 the vafxsr treNment, sectioning at eonstanl rrlslive humidity, avoiding unnexrsssry hea//n8 (sectwuss should not be strelched over a hrw plate) and avoiding eaposure of seclw.ns to light Optimal emission readings were obtained with seclNNUnt and mounting at U) W% R11 Readings were reduced by 25% when the mercury light awrce was swrtched from 200 W to 1U() W It was also 1 mportanllo kl the instrumenu wann up long enough to avoid drsfl durin8 quanluatMn Fx'h NEB should he subjecled to the same durauon of light esposurs: for a118nmer4 ( K) s) before measurint fluorescence to avoid differences frsxn phoksdecompositwn. Keirh. 1. M.. Wiky, L. A. and WI!!, J. A. //issnrilkadany 73:23)-238, 1982. Other ssff.R: College of Agriculture and Life Sciences, University of Wisconsit, Madison. From the DepsAmea>t of Veterinary Scie.ce, Univenity of Wisconsin, Madison. PLASMA PANCREATIC TRYPSINOGENS IN CNR(NdIC RENAL FAILURE ANl) AFTER NEPfIRt?CTOMY The study presented here had tlrec major objectives: (a) to investigate the relation between plasma pancreatic anionic and cationic lrypsino8ea in patients with chronic Rnal failure or nephrecqMmy, /b) to de/ermine whether hemodialysis durs the plasma kvels of these ryrrwsRens in patients with chronic renal failure (CRF) or nepluectomy, nMl Ir) u, sluwfy the elfects of nephroctomy on the plasma trypsims8:n response to Iu rtr1M N11I S, IInUI ,0l u N1 uI (qnl fC a/N' t yn1U6Cn secre11W1 in an established animal 111odel, the dn6. Fnr this study, plasma concenlralions uf anionic and cMionic /rypsino8ea were measured in C RF and anephric patients. Results showed that the plasma concenlratioro were significantly ekvated in both groups of patients. Hemodialysis did ssot cluwte thesr plasma levels. The plasma kvdsol aniunic and cuionic trypiino6esss wese highly correlated in patients and normal subjects; however, the relative concentrations of anionic trypsinosen were significantly hi8her in rewd failure patients. This su8teas that in patients with renal failure the secoat.ry clearance mechanisms for these plwna pweins clear cationic molecules mwe e(ficien/1y than tbey ckar anionic molecules. ln a related eaperiment using normal do8s, intravenous i.fwion of synthetic octapepide of clwsbcystokini. (CCK-8) resulted ilr small transiNory iwrcnta.cs in plasma irypsiao- Sea levels. After nephrectomy, bnal kvels of anioaic and cnionia trypsinu8en were ekvwed, and intravenous infusiow of CCK-t resulted iln prolonKed, high levels of plaama Irypsisw8ens. Geolas. M. C. n o!- A/werlcw lownd y/ Phrsloloey 242 (C1asuoilMesl. Uves Physiol. 3):G177~43182, 19t2. O1Aer s.yr.rti: Veterans Administration and the National lnstitswn of Health. From Martinez Veterans Administration Medical Cenler, Mrliaez, CA; Departments of Internal Medicine and Biological ChemisVy, University of California Davis School of Medicine. Davis: and the Veterans Adlnluistrationo Wadsworth Medical Center. a.d Deparame.i of SurBery. UCLA School of Medici.e. Los Angeles. I 111. HeQrt and Circulation SI.R(TfC1NIN RECEPPOR-MEDIATFD STIMULATION OF BOVINE SMOOTH MUSCLE CELI. PROSTACYCUN SYNTIIESIS AND ITS MODULATION BY PLATEI-Ei-UERIVED GROWTH FACTOR This paper reports that atrolaan, an indokamine released from platelet dese budies, stimulates pnoslacyclin (P(il,)poductinn by vascular smnuth muscle cells in cuNae through a specific serdonin-recepor-mediated mechanism. It furthct reports that serolonin and platekt-derived growth factor (PDGF) act syner`istically to elicil dramatic increases in PG1, symhesis by vascular smooth muscle cells. In the basic wait thM led w these conclusions. serotonin (3-hydroaytsyptanane: 0.5 µM aed above) stimulalod the synthesis of P(il, (as measured by rsdioinwnwwusuy of 6 kekprw- tatlandin F,a / by bovine ,urlic smooth muscle cells in culture. This efkct was atroc- lurally specifsc: a similar respnnse was nd elicited by the othea indoles or by the aminea phenykphnne. /sopfnlererNll, drqanuine, or hislamine. 11se response was revenlbk 48 1 49. 1

i and wn saturabk at semtonin concentrations of 10 pM or higher. An increase in P(;1, synthesis was elicited by the addition of a senMoeM agonist. quipatine. and ama6o- ei:ed by the seroluwun rereplor blockers eyprvheptadine. mNhyser6tde. ur methwithe- pn. The addition of Pt)GF to cultures of smooth muscle cells dramatically enhanced P(il, synthesis in respw+se to the coadminis(tation of serdonin. PDGF geatly in- creased the masimum respmse to setolottim without aherinE the half-tniaemal e((ec- tive concentration for serotoni.. Abo, this synergistic interaction was blocked by the addaion of a serdonin-receplor blocking ageM. Coughli., S. R., Moskowil:. M. A.. Aaya.la/ts. H. N. and Levine. L. Prorttdiwts oJ tM Nationof Acadrrrry of ScJenrtt of the United Smtts oJ America 79411) 71M 7138. 19t1. O/Aar saw..i: National Institules of Ik.Mlt. Fmm tAe Ikpartmenu o( Netrosurfery said Nerrobgr. Massachusetts Gener.111ospi- W. Harv.rd Medical School. Bosson; DepartmerM of Nutrition and Food Science. Masaachusetts Institute of Technology. Caetbridge; Center (or Blood Reserch, Ik- pnmrnt of Nutrxww, Harvard School of Public Health. Boaoa...d De?artment of Bwuhemistry, Bravdeis Unrversity. Waltham. MA. I HUMAN P1.ATP.I.ET-DFRIVFI) GROWTH FACTOR STIMUIATES AMINO A('ID TRANSPN)RT ANI) PROTEIN SYNTHESIS BY HUMAN DIPLOID F1BR(IBIJASTS IN PLASMA FREE MEDIA There are two major points about the action of Platekt-derived growth (actor (PD(;F) on human fibublasts that emerge from this PaPCr. First, stimulation of amino acid transpxl and /'11ikucine incorporatim are early effects of PDGF actioa on quiescent human fibroblasts Second. the sctions o( PDGF on amino acid uptake and ('H 1kucine incorporation are brought about by PDGF alone said do w require simuha- neous or subsequent preseace of plasma canPa+ew «othet haewoes. Specifically. resrks of this study show thr purified hurar PDGF induces au increase in amino acid uptake via spMem A is quiescent humr aploid RbroWasls. Cells nws( be e.prsod to PDGF for 45 wria to obub tnastrtarm tramPoA stimulation. Transport stimulnioa roauires prolcis synthesis; aansieat eapoare W PDCF, alone. M the absencM~ ~~ comParsw can stimulate lranspoA. Acid-Msohtbk ('HN~~ ~1~ stimulated by PDOF treMmeM. and thh erea( abo does no1 tequire thv: presence of plasma cornpoaents. Finallr, aalisuum a PDGF thr blocks PDCF-stirsulated DNA synthesis iw these cells also blocks PDGF-stinwlmod amino acid uptake and potei, synthesis. Increased .mino acid tptske said protein synthesis thN occur soon afta addition of fresh uxum to qtiesceM cells cr be attributed to the action of P(X;E acting aloae and should be tscful as mrtcn for Me iavesliption of early cellulu events caused by PDGF. Owen. A. 1. 111, Geyer. R. P. said Anton/odes. H. N. Proceedings of the Nainwa( Academy oJ Sciences aJ the United Stat. •s oJ America 7941011201 1207. 1922 (hhr. ir/P.rt 1/ S PLMr Ikalth Servrce and the National Cancer Lisluute I r.n. dr I1r(wrnrrM ../ NuUNMM1_ I(a1-oNd S4(tiM1) nf Public Hcalth. Boston A NGW IN)RMONAI. POLYPE!?/DE THAT STIMULATES THE GROWTH (F ('E1.1S IN ('U1.TURE: IS(N.ATION AND CHARACI'ERIZATION FROM HUMAN SI:RItM AND HUMAN PLATELETS This presentation affords a wide view of the body of Antoniades and his associ- Nes. It describes briefly (1) their early studies which led them to the discovery and isolation of a ncb hqrnrorul polrpplide growth factor (rorm htan.e serum, which appears 1o be indispensable Wr the Rnwrth o( aoratd cells in culare; ( 2) the subsequcat isolatinw and prificalion of this Folppepide from clinically ouldned hrm.n platelets; (3) the growth effects, and diverx metabolic activities of this purified PdrPeMide in cells in cuNure. and (4) studies in pror as aimed at the ehrcidrios of i1s erode of actioa in the `rmvth of normal cells in culture. In the work desaibed, studies with cell culture and a specific radwNmmunoassar demonsuaed thal 1he serwe PdrpePtide Srw.tw factor derives frons platekts and is released into serwn during blood clotting; it is uot pssenl in Plakkt • pxu plasma. This humar, plaklel-derivcd Polppeplide growth facta (PIx;F) h.a been purified to homogeneity from clinically owdaled hr,man platelets. The specific activily of tAe pr,rifkd PDGF is 20 mi0" times pmater (han that (ound in trtfraction.led human serum. It stimulates DNA replicrion and cell proliferation at a eoncawralion of I nR/ml. Characterir.tlion studies h.ve showm that PDGF is a ca- tionic. heat-subk (100'C) polypepide, with a pi of about 9.1. The wreduced rnok- cuk has an apparent moloc.lar weight of about 35.000 daNows as judged by NaDodSO,ipolracrylamide gel ekctrophoresit, and reduction results in the poduc- pow of two inactive pslypepide chaios of about 14.000 and 17.000 daNoas tespctively. Antowiades. H. N. In: Bin`, D. H. and Raseabaum, R. A. (eds.): Plaswm and Celfrdar Modulatory P•aeins, Boston: Center for Blood Research. 1980, pp. 1-11. Odet aaqPart: National Ca.ces IastiMe. From the Center for Blood Research said Harvard Uuivenily School of Public Ikdth, Bosb=. HUMAN PlJ1TELET-DERIVEDGROWTH FACTOR (PDGF): PURIFICATION OF PIX:F-1 said POOF-11 AND SEPARATRNd OF THEIR REDUCED SUBUNITS In this methodological paper. a procedure that allows the direct recovery of biologically active human p1.1ek1-deriied growth facMw (PDGF) fiom slauod gels afler separation on NaDodSO,lpAyacrylamide gel eleclrophoresis is described. This lechniyue enabled the identification and purification to horrtoReneity of two .ctive PUGF pdypeptides, ooe with a muleculr weight of about )3.0(10 (PIX:F-I) said the other with a molecular weight of about 12.000 (PDGF-Ill. Reduced PDGF-1 produced two i.aclive subunits, with molecular weights of about 15.000.nd 13.000. Reduced P1 X iF-II alsn produced Iwo inaclive subunils, with molecular weights of about 1 S.0(10 and 16.(KK). It is pnsibk that PIX;F-II derives fuMn poteulytic cleavage of P1K;F-I. ,t, 1 51

Anaviiwler. ll. N. Pna-eedinRr aJ the Nuriww/ AcuJew.y, oJ Sriences of the Uwirrd Srarrs u/ America 79412) 7)11-7)17, 1991. OOther sryp.+f: National Institutes of Ileallh. From the Center few Blood Research and Departmenl of Nulritiun. Harva d School of Puhhc Ikahh. Boston. ST1M111-AT1ON OF PIIOSPFKxJP1D AND CHOLESTEROL ESTER SYM711SIS BY Pl.ATFLET-DERIVEDCROWTH FACTOR IN NORMAL AND IK)MOZY(i(H1S FAMILIAL HYPERCIK)LES7F.ROl.E7NIA HUMAN SKIN h7BR(XILASTS The aim of this investiaMiow was to study the effect of human plaiekt-0erived growth facwr ( PIX:F) on hpid nrubo/ism, partinlarly choksterol ester und lirospho- hpid synthesis in nornsd and familial hypen:hoiesretolemic-0erivod Iwnion skin fibroNaas. Resohs of tJws study show M.r pure PDGF at arw`ram kvels stimulates chMdesrerol euer. photpAoliprd rd DNA synthesis in bodt normal and familial hyper- ehukstemlemu muud human skin fibrvblasts. While stimulation of DNA syMhesis did nol begis wwd 15-24 M after addition of PDGF to quiescent norsnsl and FH mulw fibrvblaus, stimulatan d('Hiokic acid incorporation into chokstcrd ester and phos fiholipid was evdeM ) e Ms after the addnion of PDGF. In the nrxmal cells, dre rare of cM,kstefd esocr symhesrs was masimal at 24 hn . Ihen rapidly declined. ClKrleskrd estenficMion was owch lower in the FH cells thaa in the normal cells. The stimulatM,n of ('H/okic acd rncorpnntan into choksterol estes by PDGF was inhib- i/eA is both normal and FH mutant skin 6broblasts by pro6esterwrc. an inhibiwr of acyl-CoA: choksierol acyhtansferase. The rate of chokslesol esler synthesis in the normal cells increased as the concestratioa of pWekt-poor plasma or low densily Iipopnwci. (LDLI was wrcrenod. espocially iM 1he p><sence of PDGF. Lincarization of the LML dose-response curve indicated that PDGF iecreased the rate rather than the affinity of the overall chokslerol esserikrion system. The rate of choksterol esteri- /kation in the FH mwant celb was highesl is 1he absence of LDL or at low kvels of platekt-pnor plasma. Consequently. PDGF can stimulate eholesterml ester synthesis by U)L and aon-LDL-wredissed processes. l.eslie, C. C.. Anron/adet, H. N. and Geyer. R. P. siocAi,wica n eiapAyaka Acso 711:290-)01, 1962. Fraw the f?eprtment of Nutrilio.. Harvard School o(Public Health. Boston. MIGRA7ION OF Ctll_Tt/RFD VASCULAR CELLS IN RESPONSE TO PLASMA AND PLATEIJsT-DERIVED FACTORS In the study presented hent, a quantitative assay for cell milraMN+ was used to measure the R.rveMse of pencyles, smooth musck eells snd enddhrlial cells fmm lar6c and small vessels w human serum, plasma and puified platelet 4.enved facwrs. In Ihe first pars of this sludy, phasokinelic mi6rYion of cultured vascular cells was tested in resprose to human plaekl-rich serom ('serrm') and human plMekt-poor pla ima serum ('plasma't. The cell types tested included bovine sonic endodrelid cells, human umbilical vein enduwhelial celh, human hemanKiomal capllary e.ddhe- lial cells, bovine aJreeal micrw,ascular pericyles, and bovine .ortic smooth muscle cells. Human serum slimulaled a si`nibcaa iaeease i.1Ae rase of miRraios in all five cell types. Human plasma stimulated drc eadoqKlial eelh so mijrale but hd no effect on the migration of pericyles or smooth muscle eelb. In dre second psn of this strdy, highly pwified plauekl-0esived Krowth factor (PDGF) sliasrlaled dose-dependeal nr- Rrauon of srnooth musck ceNs causing a 50% increase in ph.Roki.dic track area relative 1o cnntrds. Neilher pericyle aor eadwheli.l ce8 migruion was slimYlnod by PDGF. Rabbit amiserrm to Iwnnn PDGF complaely blocked dre smooth nrasck cell migration induced by either 10% serswn of h>t/ml pure POGF. Purified platelet lactor 1 V( PF.) stimulred misration of ptricytes but .ol of smoolh swsck ceBs or e.dothelid cells. Sheep antiserwn 1o humsn PF, eornplelely blocked die pericyte migration ia- d.ced by eiaAer 10% aeruwr or 1 uR/nd pure PF,. TLese results indicate that PD(iF is dte primary facwr in srnae n:sponsiblt: for the migration of cuhwed aortic smooth muscle cells and Uuw PF, is a critical fanor required so induce dte miRsatioe of pericytes. Other faclors present in both pl.sma and serum conuol the saip.tiow of vascuhY endodrelid cells. Berwstein, L. R., Anroniadei, H. and Zeuer, B. R. /n+vwaf o(Cell Sctence 36:71-12, 19E2. Otfiee sapP..t: National lnsdlutes of HeaMh. Fvm dre Deprt"'enu of Pltysioloty WA*SwRery. Harvard Medical School. CiU- dse.'s Medical Cedu, and Harvard School of P{rblic HeaMh, Ceswa tor Bk+od Re- search. Boston. PLATELET-DERIVED GROWTH FACTOR BINDS SPECIFICALLY TO RECEPTORS ON VASCULAR SMOOTH MUSCLE (-'E1.(S AND THE BINDING BECOMES NONDISSOCIABLE Plrekt-derived growth facsor (PDGF) is a potenl stinmlam of the pos+nY of vaaculr snaoth musek.ln order so wrderstand Me action of PDGF. mahods had so be developed feu identifying PDGF reeeptor sites. To aeeomplisb rhis, sadioiodiwred PDGF ('"I-PfXiF) was used in studies of PDOF binding siks on vascular snsooth nwsek ceRs. There was an escelknt cbnelatinw between qrc ability of "1-POGF to stimulate cell prolifcration and to bind specifically 1o euhurod vasetrlar smooM nusck cells. The hap-maainul coneenlration for both processes was 0.1 aM. There were 30,000 binding saes per cell. Reduced PDGF. prepared by Ireaunen~ of PDGF with 20 mM d'ahiahreitol. had sKidtet the ability so bind to smooth aawck cells nor to suinw- t.1e cdlulr proliferatioe. Epidertnd growth (attor, nerve growth fartor, 6broblasl growth facu>r, and hisaMe B did mrt compete for da binding sites M a concenassion of 10 aM. "I PDGF binding was skrwly reversibk at 1•C and was rapidly and totally 52 1 53

I reversible aftcr a I min incubatMin M 71'C. After continued incubation at )1'C. the ' Mndrn6 brcan,e rrrcvenrhk. The half-bme fot frxmation of the nrrnrhsuxiahk slate of *"I-PIX;F was - S mrn at J7-C The nnndissociabk state ol hindmg was mM Irwmed at 1'(' even a(ter I hr of incubation. These data suggest that the labekd rnes are the P1X;F receptors that mediate PU(;F's mitn`enic action and that a nondiss,tiraMe state of PIX;F bindrn` is fo.med at 3rC. It is likely that nondissociabk 1'tx;.= rcpresents internalired Irgand or binding to siles IAM are eonverted to a high-affinity s ate after the Ibgand brnds. Williams. L. T.. Trembk, P. a.d Ansow/edrs. H. N. rrorccdings of the Nariono! Aradrwy of Sclewcrs of rAe United Sraaer of Arnerico 79 SfIb7-Sa70, 1982. t'JtAcr srK.rr: National Instilutes of HeaM and dre Mihon Fund of Ilar,rd Medical Schoul From the Cardiac l)nit, Mns.clwsetls Oe*eral Hospital. Boston. and dre Center lot Blood Research and DepartmeM of Nrerition, Ilarvard Univenily. Schonl of Public stcdth, Iloston. I C()NiRIBt1Tl()N OF PLASMA PROTEASE INHIBITORS TO THE INACfIVATI()N OF KAI.I.IKREIN IN PLASMA This study was designed to usess the respective conuibution of the differed plasma protease inhrbrkxs to the inactivation of kallikrein in plasma. The use of a new technique that selectively inactivates e,M in plasma frcilitated the comparisnn of the kinetics of kall/kran inactivation Resuks showed that, under pseudo-first-order eon- ditions, the inactivMion rare constaM of ktllikrein in normal plasma was 0.60 mi.'. 'ilris rale constant was reduced to 0.35. 0.)0, and 0.06 min', in plasma deficient respectively in Ci-INH, a,M. or both itrAibiton. 71ws CI-INH (42%) and o,M (50%) were fornd a be the rnajor inhibitors of k.lliluein in normal pfasma. Moreover, all the odher pwease inhibilors preseM i. eoret.l pla,snu contribwed only for 0'1ti to the inactivatioe of the en:yme. To eonRnn lhese kinetic resuNs,'"1-Kailikrein (M. a5,000) was completely inactivaied by variow plwna sampks, .nd the resulting mistwes were wlyned by fiel fiNnliot on Sepbnoae 68 CL for the sppearance of '"1-kalli- krdn-inhibitor compleses. Overdl, drc res.Ns pesentorl here i.dicate tlw C1-INH and u,M are the wujor irAibiton of kdlikrein in sorreal human plasma. This conclusion is srpporned by (a) die analysis of the kinetics of kallikrein auctiva#ion in mxm.l awd pvkase inAibitordefkiewt plnat. snd (b) by the,wantilstion upon ge16Nra1ion of the kallikreiw-inAibitor compkaes formed is plasma as dre tesuh of the inactivation of purified rs0iolabckd en:ytne. Schapirs. M.. Scod. C. F. and Colwwn. R. W. lownof oJCGnicof lnvrrrigorion 69:162-465. 19d2. OtAcr sapP.rr: National Insututes of IkaNh. FFn,rn the TLnHnM+an Rescreh Center, Tempk l)niversity School of Medicine. I'hrladclphu H1GH MOLI:Cl1LAR WEIGHT KININ(X;EN OR ITS LK;IIT CHAIN PR()Tl:('1'S IIl1MAN PI.ASMA KALLIKREIN FROM INACTIVATI(Nd BY PLASMA PROTEASE INHIBITORS Five plasma pnNease inhibitors ue known to inanivate kal5krein, but now i1 haa been shown also that high molecular weight kinwwaen ( I IM W K) or its light chain can act to protect kallikrein from inactivation by these inhrbitors. For this biochemictl audy, the kinetics of the inanivNinn of kallikreit by a; m.cro`Iobuli., an1i11nonrbin 111, and a; antitrypsin were noted in the presence ssd absence of HMWK. 1n the absence of IIMWK, the second-onkr rate constants t.,/K, for t!rc isactivalioo were respeetively 6.9 x 10', 1.8 x 10', aed 2.5 x 10'M 'min''. Whew HMWK wu pn:sent during the rcaction, the inactivMion raAes by these pfasmm prplease inhibibrs were rcduced as a resuN of the Cormation of kallikrei.-Migh ntokrular weight ki.iworn compks. K. a 0.75 pM. W(tcn the light chain derived fran redretioe of kinin-froe IIMWK was suAstitwed for the p.rewt nrdecrk, a rnore; pronounced reductiow is inac/ivMan rares was observed due to 1Ae fmwryiow d a kdlikrein-IigIM ehun conipka (K. - 0.11 pM ) These resu;ts dctnonsUate qry 1he combining site for kallikrein o. IIM WK, responsibk for cornplea farnration and prvlocyion against irrhi6ilan, residn i.1he light-chain reRiow of tke molecWe. Mareover, kallrkrein appears to bi.d rnone ti6lNly to the light chain of HMWK 11un to the parewl molecule. Schspr.. M., Scott. C. F.. l.nles, A.. Silver. L. D., Kreppers, F.. lamcs, H. L., and Cofawn, R. W. SiocAeiwisrry 21(l):367-372, 1982. Qf4r sstpp..t: Nrion.llstinMes of Ikahh. From the T7rombosis Research Center and the Dep.runcnt of Medicine, Tewple Univenily HeaNh Scie.ces Cewter, Philadelphia. REGIONAI. RENAL AND SPLANCHNIC BLOOD FLOWS DURING NWOTfNE INFUSION: EFFECTS OF ALPHA AND OF COMBINED ALPHA AND AETA ADRENEtO1C BIAC'KADE This study was uwitwed for the purpose of dr:fining ddrectly the rdes o( .IyrA, adrenerRic and pu.sibk tsoesdre.ergic tnechanisnu in vasocunstricsor re.p.w.es as nicotine. Todo tAis,.icaiae-inducedcAaKes in blood Bow awd vascwlarcondrey..a iw rsEiond renal and spl.ncheic beds were evalrned before s,nd afler (l) selacdw biockade of *b adrenerjic receptors and (2) combined blockade of.(PAf sad iwW adrener6ic tsceptors. Before adrenerlic blockade, nicotine brcreased anerid prewra (+ e2'!<) but had he/nogeneors direetional efkcts on regional blood Oows: p.ncreas (- 6496), dunde.wn /- )l%), kidney coAea t- l1 %), kidney aneAuqa ( - 171k), liver (+ 3%) and sploen (+ 71%). Vascular eandrer.nee was sedrrcod in kidney eortes, kidney medalla. doodeswa, liver and pancteas, swd was.ot aMered in spken. Selec- tive elpAo adrenergic blockade ptevenred she hyperknsive response to .icotine, but Aetengenerws chan`es in n:diond firws persisted. After combined a4Ae and brre adrener6ic blockade, hicotine increased syskmie anterW pessure rd docteased 1 54 1 55

vascular coeducturce in all tissues. Resuks indicate that there are (I) n heterogeneous influence of nicotine in renal and splanchnic circulations associated with regional differences in activities of alpha and beta adrener8ic recepton and (2) a pitent non- adrenerRrc vasocunslrickx response ia these cirtulations to nicoline after bMxkade of afPAa and beta adrener8ie receptors. Downay. H. F.. Crystal. G. J. and BaaMar, F. A. TlvlnwwaloJPAa.w.ocoloayanlEsPeriwrntaf Tbr.apewics 220(2) 375•381, 1982. OrA.r z.yPwrr: Cardiob8y Fund. From the Depvtments of Physlolo8y ssd lnternd Medicine. Uni.xrsity of Tesas IkafUt Science Center and Cardiovaacrlar Researeh Laboratory r Meahafist Hospital, Daflas. MFCT OF CIGARETTE SMOKING ON HIGH DENSITY UPOPROMN PHOSPIIOUPIDS Tfus study was designed b aors.lhe effect of srule inlulMroo of ci8arette smoke on hi8h denury Irpopruran 1HO1.) phospholipid compoailio.. 1s rurnin8 the testa o. Whre Caraa" pi8eons, the rolbwi.8 rorr treameM re8imens wen: iwituled: (1) sheN control birds (ed a chow dai aed retained in their ea8es: (2) skrn pigeons fed a choksrerol-utuwned fat drct and esposed so fresh air by a smokia8 rnrhine• (3) low nicdrne lo.w carMn nsonoarde (l.ol.o) animals also fed the ehoksierol-fal diN and e.posed to low concentrations of these ci8rette amoke products; and ~ 1) hi8h nicotine- hr8h carhon morrnide tHrlb) birds fed the choksterol•fal diel md subjecled so high concentrations of these idulanu. Resulls of these studies shuwed thss the cholesterol diet caused an increase in the concentzation of most HDL phospfwlipid classes. Eapo- swe 1o the HrHi regimen resulted in an increase is the HDL choksterollphospholipid ratio and a reduction in the concentration of HDL plasphnidyl ethaaolamine. Phos- phMidyl aerine/inositol, lphin8omyelio rd IysoQhosphatidyl choGne. It appeary, therefore, that ci8aretle smoking may a+peauue HDL's ss+tLatberr><cnic properties by dtenng surfsce plwspholipid components. Ikprty, K. M., Tureiss, L. E.. Mullip., l. ).. Cluette, l. E., Kew, R. R.. Suck, D. l. , and Hoj..orl1, 1. L. QiocArrnicaf and AiopAysical Research Cor~nnnlcations 10i(1):212-219, 1982. Other z.w.rt: American Ikrt Associstlon, Greater Boston Division. Frvin the Department of Biological Sciences. University of Lowell, Lowell, MA. AI.TI:RNATIVE COMPI E:MENT PATHWAY-DEPENDENT INGESTION OF I1.lNN fTf PARTK'US BY HUMAN GRANULOCYTES Pan.. k...l a 11u.w<s.rnr i y( lic hydrrxarbr» /Flurrlue) have bten relwwred before ho t.c rnctYnl by RrNMMuI t( Y pha8ir.ytes of human blood This paper describes I quantitative studies of 1hc in8estiun of Ihcse sanr particks by human Branulocytes. In the studies presented here. Ruotile particles with an average size of 0.1µM were ingested by human Branulocytes after incubation in fresh normal human serum. InBes- tion, which was assessed by visual counting in a fluorescent microsurPe of cells containing particks, reqnired fresh normal serum and did not occur when serum was healed (or 30 minutes at SO'C or when ethyknediaminetetra.cetic acid (EDTA) was present. Particularfy, it did not occur in serum genetically deficient in Clb insctivror or in C3. However, plu8ocytic activity was restored to C3-debcieat serum by purified human C3 and 1o heat inactivated serrrn by purified factor B. Opsonization of prticles under appropiate conditions is insensitive to the absence of hwnan C2 a CS, but is dependenl upon C3 ard an nNacl apenwive C PNhway. Meas.remenl o( Me opsoniza- tion of these particks thus constitutes a simple assay of dre functional irNegrity of this pathway. Although the mechanism involved here is slip wrckr, this simple and a.dily, available assay can be used clinically n. snocw for the functional opaonic activity of the ahern.tive C pathway, r well as for Ihe capacity of human Branuhxyles b reco8nize ard ingest C3•coMed prticks. Ar.aorM, M. A.. Lwciaskr, F. W., Lionrtri, F.1., Alper. C. A., and Vakri, C. R. The Journal of Inunnnolojy 127(1):278-281. 1981. ~ Osher s.pp..t: Of&e of Naval Research. From the Division of Cell BiobBy and NepMokKy, Children's HospiW Medical Center. Boston; Cenar for Blood Research. Bostw; and Naval Blood Research Lso• ralmy, Botlon. ENDOTHELIAL METABOUSM TUere are three importrN fwrctions of endotheliunt-(1) sepr.tion of blood n.d lymph from the estnvrcrlar spt+ce.(2) control of the idlux and efMn of specific bfood sotutes and colloids. and (3) provision of a smooth gliding surfaee for Passi+g bload-dw have been well recognized for years. The book chapter presented hem, tlrou8h, deals not with these tlree fwrctioru bu1 with the melabolic activities of the ea6wheliwn. While endothelid cells are we8-eqripped for Blycolysis, osidNive "_ phorylalion and the compks series of reactions reQuired fa cell divisia.. N a the number of far reaching metaMrlic auivities, mosf discovered since 1968, dul are cuesidcred here. To wil, endulhelial celis possess a highly campka array of irwracelhr- lar machinery and can, in (act, elaborate molecwks.ed macromolecules important not only so cell integrity and cell division bw also important in tanrs of processing vasoactive and hemostalic substancei. Many of 1he recently recognized rnnabolic activities occur at rr new the cell surface. For esample, anBKMensin•convertin8 en- zyme appean to be disposed so thN the enzyme itself is embedded in the PlasmA membrane, yel its catalytic site is situated to have access b angiolensin I and brady- kawn as they pass in circulaMinB Mard One section of this psPer deals with.nromicJ eonsiwkrYions. arwf anulha with surveyin8 the adaheliuni s km)wn metabolie activi- uea. While much of the wurk on endrAhelial metabolism per/urmed so tr has been accomplished through the disciplines of birahemistry. narPhuGgy, and cell biology. 56 1 57

other studies. Physiobwric. phnmac.logic. and pathologic. are fcasibk. and there is every rczwn to believe that clinically rckvrN data will be furtlKUnung Ryan. I W anJ Ryan. v. S In: WieJman. M P(ed 1 An Introdreriow oJMicrortri rlmiirn. New Yrwk: Acadcmic Press, Inc 19111, pp 117-169. (hAer nqPert: 11.S Public Health Service and the John A. Hartfrxd Fuundatwn, Inc. From the [kp+rtment of Medicine. Univretsily of Miami School of Medicine. Miami. FL. STRUCTURAL BASES FOR METABOLIC ACflVITY The pulmonary endrnhehum. which is the flrst line of contact between blood- bnrnc srbtrres aed the «llular machinery of the hm`s, is reviewed here along with thwe structura,t specializ.tions that we suNed for Iheir rok in the meubouc functions o( the bngs. The underlying structur.l bases for metabolic activity are considered hae under the throe headings: anatomical, eellular, and subcellular. Particular emphasis is placed upn the enzymes, inhibiwn, receplors, and transport mechanisms lhat ne impowud in the regulation of the hornwnal eompo+ition and Aui&y of blood. Fdbw- in# anatomical study. a pictwe emerges of a variety of enzymes that are situated on the surface of prfmonary endothelul cells a.d we wnegically poised (or interacuon with the aqpropute substntes delivered by the blond and are equally strasegically Placed to dcyermine the quantities of active sub.unces allowed to pass downstrcam or into the estrsvascular space. Cellular studies show tlut the cavedae-plasma membrane frac- Iion of htng hartogtaate converts anSio/eruin 1 a angtoteesin 11 and degrades br.dy- tinin to yield the characteristic products formed by intact hmts. Furthcrmtxe. the preparalionun degrade ATP and S'-AMPto adenosine and troe pAosp*atc. By use of a .ensitive rsdinassay, it was also shown Ihr pulmonuy endothelial cells contain abun- dant anSiolensin converting enzyme (ACE). let wmrnary. it appears that the unique ktltres of pulmonary enzymes may re1Me to the atrunwe of the has`s and to tAe position of the lung in the circrolatory system. T1ws, the 1ota1 amowM of ACE within the Iwp may greatly eaceed the amoud aeeded 1o process the concentrat on of anaiwen- sin I and bradykinin tnually found in parlmonary artery blood. While it is knowa that ACE uccurs on the huninal surfaoe of thesecells, it is becomin` eviden a1 a subcellular kvel that the membrane bound enzyme is sensilive Io factors such as osY6en concen- tration. However, the effects of components of the plasma membrane c.e the molecular con6gurstion aed hence on the activity o(1he enzyme we not known as yet. Ryan. U. S. Annrrd Review oJPAysiolm 41:22)-2)9, 1982. (kAir s.wwr: National Institutes of Health. Fn,m the Ih/*.rtmenl of Medicine, University of Miami School of Medicine. Miami, 1•1 I NI:UTR(WHIIS ARE REQUIRE[) FOR THE DNA SYNTHETIC RESPONSE OF HUMAN LYMP4HlCYTFS TO MEVALONIC ACID: EVIDENCE SI K:(:1?STIN(i THAT A NONSTEROL PRODl1CT OF MEVAf.ONATE IS INV(N.VI:1) The ability of various enantiometic forms o( mevalonic acid to initiate lympho- cyte DNA synthesis is eaaminod in lhis pper, and thc role of dtc neuuvphilic Poly. morTlw-kar kukocyte as the helper cell in evoking tnevalonate-induced IynPhocyte pruliferation is defined with g realer precision than has been acconmplishod bcfore. 71te ability nf lymphocytes to inNiate DNA synthesis and cell cycling Is • radiosensitive poperty of the cells. whereas the helP provided by netrtropAils is main- tained despite their prior eapnure to a-inadi.tion. Otha organic acid.niows, iwck+d- ine precursors of ineva{onic acid bin.ynthesh and a variety of products of mevak4nase metabolism. fail b Mtitia/e DNA synthesis when added b human lymphocytes. Be- cause only ahe meubolicallr aclive RI -) en.ntiorner of inevalwtic acid InNiues lymphocyk DNA synthesia, N.eenn probable thM physlologkal pathways of ntevalo- nae metabolism are Mvolved. The tesponse ro mevalnnic acid of ML-236B (cornpac- ti.)-inhibited lymphocytes is incaased, and the lhreslydd concerMratiow of inevdansle at which lymplwcyte DNA synthesis fast appears is docreased, when the oepa we cuhured in lipoprotein-containing medium. The response 1o mevalonic acid of lyrn- phocytea cuNured in lipopaein-depkted medium can be enhanced by addition 1o the cultures of low density lipoproleiu but nol by addition of high density lipopalei.. Based upon the Bwt diversion hypodrcsis of inevalona/e sneubofism.lhese observa tiar suggest that a nonstaol pmdutt of inevdasre metabolism may be resporuible for the initia/ion of lymphocyte DNA sys>thais by mevalonic acid. Lsrson, R. A.. ChunR, l., Scaw, A. M., awd YacAnin, S. Proreedines of rAr Nriowaf AcsJewy of Sckncea of rAe (Inited Saor.a oJ Awn(n. 79:3022-3032,1982. OrMr aarp.rt: U.S. Public Health Service a.d t6e Nalco Center Research FtMd. Ftom the DeP.rtmaNa of Medicine asd Biochemistry, the Franklin McLean MetworW Research Institue, aoad the Comenipoe on Immunology. The University of Ct+icyo School of Medicine. Chicago. MEVALONIC ACID IN CONIUNCTION WITH HELP FROM NEUTROPHILS INVUCES DNA SYNTHESIS AND CELL CYCI.IN(: IN HUMAN CERIPHERAL BLO00 LYMPHOCYTES Mevalonic acid plays an important role in the regulation of nsarswnaHan eeN growth and division. Results ftuonearficr studies have even suggested a csitical role (or mevakonic acid, independent of Ns convenion lo cholesterol, in the regulation of DNA synthesis and cell replication. Evideonce presented in this paper shows thr mevab.ic acid does stimulate DNA synthesis idhuman peripheral blood lymphocytes which h.ve been isolated by gravity sedimenution of blood and freed of adherent cells by nyksn column passage. Ituman peripheral blood mottt>rwckar cells isolated by the Ficoll- Ilypsyue technique respond less well, but their response so mevalonic acid c.n be enhanced by the neutruphil•rich Ficoll-Hypayue-iaolated "batom" cell fractiuw. The kiacctics of inevationic r•id-induced lymplwKyte transformation are siwtilar to tho.e of nrne classic lympfwocyte miNjens In additwro to sumulalin` lynplacyte DNA syn- Ihesis, mevalonic acid pndures a prulaiun o( cells rcpresenting all phases of the cell is 1 39

cycle wtwo.e nwapfwrMgical characterislics are typical of thosc sccn with tmrre cnnvcn- tional min.r6ens . Tlrc 1)NA synthctic response of lymphocytes to mevalo ac acid can be abo.lishcd by prar eannure of the lymphocytes to s-irradialion or miwnsycin C, while the helper effect uf 6ranukrytes is unaffected by either treatment. Thesc observations suggest that mevatimic acid mi1 only may paay a rok as a critical su-fttance which supfxxss the prufx>6alion of cells protranrned to divide, or stimulate7l Io divide by various initiMcxs of cell srowth, but also 1hM, in susceptible cell ppu)alrons, meva- h,ntc acid may act as an inducer of the entire program of the cell cycle. YocAnin. S. and Richman, D. P. ('clfulor hwrnunoloty 72:21e•262, 19l2. Other suPprt: 1/ S. Public Ikahh Serviee, Muscular Dystrophy Assoctation, and the Nako Rcserch Foundatinn. Fnwn the flepartrnenl of Medicine and Neurology, the Franklin Mcltan Memorial Reseanh Inslnutes, and the Cnmenince on Immtrrobgy, The Ilntversny of Chicago Schnol ol MetlKine. CTta'a6o IV. Neuropharmocology and Physiology NICOTINE BINDING SITES AND TIIEIR LOCALITATION IN THE CENTRAL NERV(x1S SYSTEM The study of nicotine binding in the brain, which was plagued with Probktta during earlicr investiaatirun. has been helped greatq by the development of a method of obtaining the unnatural ( f)•isortxr optically pure and by the refined synthesis of radiolabekd nicotine with high specific activity. The actions of the slereoisomen of nicotine on the central nervous system are audiW ively similar in mosllests bu1( -)- nicotine is mae potent than the unaatunl (+}isomcr by at least 10-fold. Binding of sadiolabeiod nicotine to brain has both saturable and nonsatrrrabk compooeats. Only saturable binding is affected by incubrion cowditions such as time, tcmperatwe. PH and ion cuncentration. Escess concentrations of the sleredsomers are eyually effective in displacing ( - )-l'HI•nicotine from brain homogenates. Neverthekss, a direct can- p.rison rr( (+)•/'11) •aicotine and (-)-1 *11-nkaine binding shows that the latter has a K, thnee times bwer than the forsner. (-11~11•nicoline is bound to the greatest de`ree ; in hypothalamus and hippocampus, ase.u that also eshibit the must stere oselectivily for nicotine Ibwever, differences in the binding affinities of the Iwo uor sers arc fr less than the pharmacological stereospocificily observed. Martin, B. R and Acero, AI. 0. Nrwosr icnrc d Rchavia.a/ Rcviews 3(4):173•175, 19141. OIAWr srpp+rt: Nitwmal Institutes of Ika11h. I r..rn dr IkpwinrnM .d /'hunur o/ogy, McdK.l ('ulle6e u/ Virginia. Richnwurd I C'l1ARACTI:RIZATION OF NICOTINE BINI)ING IN M(H1SE BRAIN AND COMPARISON WITII TIIE BINDING OF a-BUNGAROTOXI": AND QIIINUCLII)INYL BF.NZILATE The literawre suggests 1hN nicotine binds b neurond tissue, but the nature and significance of Ih/s buWuK are in question. In this study of nicotine butdsng, the bindingof /'II IniuMine to snouse brain was meastued and subscqueatly comparcd with the binding of l'lla-bungarolosin (a-BTX) and L-I'11) quiwclidinyl bentilale (QNB). The binding of nicotine was saturabk, revenibk and stereospecific. The averase K, and B,,, were S9 aM and Ita fnwrks(etR of protei., respectively. ANhough the ra/es of association and dissnciaion of nicotine were temperaUredependenl, the incubation tcmpcraure had no efkcl on either K, or B_ When measured at 2a' or 3T, nicotine appeared to bind b a single class of binding siles, but a second, very low- afhnity, binding site was observed at 1'. Nicoaine binding was unaffected by the addilion of NaC'1, KC.'1, C.CI„ or MgSO,1o the incubation medium. Nicotine cho- hnntic a6neists wcre Puteno inhibitors of nicotine binding: however, nicaine rMago- Msls were poor mh/biaxs. The regional dtsurbulion of binding was not uniform: rnw/hratn and sniatum contained the highest number of receplors, whereas cerebellum had the fewest When nicotine, a-BTX, and QNB binding were compared in seveal ways, results indicated that ell tlttee li6ads label choli"ie sites, but these sites differ from nne another. Differences in site densilies, regional distribulion, inhibilor poten- cies, and thetmal denahrNion iwdicalod that nicoune binding was no/ the same as either QNB or a-BTX binding, and drcrefore that tccePlort fa nicotine may represent a.niaNse population of choiinesRic seceptas. Marks, M. ). and Collins. A. C. Mol4cr/ar PAarnmcoloty 22:S34-S64, 19E2. Othtr srN.rC National lastituks of NeaNh. From the Inslituk for Behavioral Genetics and School of Pharmacy. Universil7 of Colorsdo, Bwlder. EFFFCTS OF ACUTI? CENTRAI. AND PERIPHERAL ADMINISTRATION UP NICOTINE ON HYPOTHALAMIC CATECHOLAMINE NERVE TERMINAL SYSTEMS AND ON TNE SECRETK)N OF ADF1rO/1YPOPNYSEAL HORMONES IN TIIE MALE RAT The actions of in/nveMrkuhr injectww and intravenous infusions of nicdine were evaluated nn daQ.mine and mnadrenaline staes and Iurwover in discrete hypoth.- lamic rkrpam®r and woradreealiee norve terminal systems in male Spasue-D.wky rals. Anterior pituilary hurnsone secretion was dso studied in the same group of animals. Specifically, meastmements were made of L", FS/I, GIl, TSN, proladin rd caticastcvune serum levels using radiuitsttr..toh>tical procedures. Inu.individud carrelalions between resiond caterMrlatnite kveb and Mtrnover and honnone secre- Iion were perfixmed. Results showed that iMraventriculr injections of nicotine pro- duced dose-dependent reductinns of dopamme and noradrenaline levels ard increases rf dopamine and mwaJrcnalme turnover in discrete hypMhalamic areas The turnover ra1 1 61

I- increases were associated with reductions of serum levels of TSH, prolactin and 1.11 anJ .n increase of serum FS11 levels Carrelatran analysis in the intn ventricular eapenments was pcrfurmed. Also. intravenous infusiwms of nicotine uvr r one hour produced dose-delsendent reduction of duparnine and tsuraddrnaline crxrcen 'nuons and increased the 1wsNrver of these moooamines is the variews hypothalamic catecho- lamine nerve tenninal systems analyrsd, with the median emincnce, da,»mine and rradrenalrne nerve terminal systems showing the highest sensitivity to nicotine. Overall, the total of these and other eaptxinsenul findings reinforces the view thN nicutine may act daectly un the brdn to attisrale dopamine and noradrer aline nerve lermuul syaems, pn,bably via activalion af aiootine-like cholinergic recepwn. Corse- lattm analysis shows that othet neuratra.n.Mler mechanisms must also be involved in pnducarg the wicdine-induced ch.nges in Ihe tiecretion of anterior pituitary Irsrmones. Andenson, K., Fu.r, K., EnesdA, P., and Agnati, L. F. Mrdicaf Aio1oRy 60.99• 111, 1982. lOther srqp+t: Svenska Tob.ks AB, Stockholm. From the Ikprtnsent of Histology, Karolinska Institutet, and the Hormnne /abrxa• Lwy, Kantirnsla Hospital, Sksckhohn: and the Department of Human Physiology. Wivenny of Modcna, Mriden., Italy. ON T11E INTERACTION BETWEFIJ NICOTINE AND CYCLOIIEXIMIDE Inhrbrton of protein synthesn, such aa cycloAesimide, have beea shown to affect memory retention and nicotine has been shown so reverse the amnestic pnrperties of eyclohesimide. In the study presented here, the interactions of nicotine and cyckhea- imide on brsia protein synthesn were esamined in an attempt to determine whether the nicotine-induced rcvenal of the arnnestrc effect of cyckrhesrmide is due to an interac- tirm between nicotine and eyck,hestmide on brain protein synthesis, aed whether it involves the sNes in brain that saturably bind 1'HNticoline. Results showed that nicotine did nor reverse the cycknAesimide-induced inhibition of protein synthesis, both in vivo (intact aainul) and in vitro (bra/n dices), suggesting that on-going protein synthesis is not nocessarily invdved in mernory consolidation. Also, the nicotine binding sites were not affected by In rivo or in vitro treatment with cyckshesimide either in the presetwe or absence of nicotine. Sershcn, II., Reith, M. E. A. svd LojrAa. A. Aroin RnrorcA 231.IS3-1113, 1992. From the Center for Neurochcmistry, Rockland Research Institute, Wnrd's Island, NY. auempt to Ies1 the hypothesis that nicotine acts as a ttceplor (known or unknown), the specific binding of'll-(d,1) nicotine to kukocytes was measured. Results of this study denw aauate the presence of a nonnchulinerqic nicotine ticeplor on Murun phasocytic kukocyles. The average dfiry/y •/-standard deviation of (d,l/-nicotine fot 1he rocepNK on ncutrophils is 36 •/- Id aM (n=6). The binding is saturable with an average of 9.7 a 1l?' sites per neutrophil. Motntyles and, to a ksser ealent, lymplacy- 1ea but ma erythrocyta also display specitie binding. Bound nicotine is dissociabk from the receptor and is not metabolized. Only close strnclural andop of nicoliwe bind to tAe nxeprr, which is skteoamlective for the (d)-isowrcr. Ttte seceptor can be occu- pjed by (1/-nicotine at conceswations present i.1he blood of smnhers. It is suggested thas some of the adverse effecla of smoki.R on kukocyle fwrctioss nuy be modiMed by a specific nicotine receptor. Davies, B. D., Ilors, W.. Lin, )-P., and LionNti, F. Mof.cular.wd C.llalar elocAnwbrry 11:23-31, 1962. OrA.r sqr..f: I). S. Public Heahh Service. Fran drc Cenlu for Brain Research, Universily of Rochester School of Medicine and De.tistry, Rochcsscr, NY, and Center kr Blood Reseasch, Boaso.. CHARACTERIZATION OF THE ISOLATED PERFUSED MOUSE BRAIN AS A SYSTEM FOR NEUROCHEMICAL STUDIES The criteria for characteriration of isdated petftued brnin preparMiowm i.chde 1he demotmratioa of ekcticd, sancttral and seelabolic viabilily. The purpose of tk,ia study was to meet tltese criteria and b tslaMish The kmlrr.d perltsed atouae br.i. (IPMB) tl a viabk model for nerrochemicaJ and wxwopArws.eolo~Kd studies. lal dte paper presented hexe, the preparation of the IPMB is dactibed along with iu tkx- dophysioloRical. marpboloRieal. biochemieal aad plnrmat:oloRicr properties. Using wth peda*neace liquid chromatograPM' with elecuoclatemicr dNCctios, the priary arctabolite of mammdi.n ceawal ncsvoraa systam worepnepMwte, 3-metlway-l-hy- dtvsyphenethykae`lycol (MHPG). wr measttred is the perfusate at 13-min. itrer- vals. The rate of MHPG ptoduction was dnwlar to Ba~attn: values of the rase of •orepinepMine ttuwover is mnrse brain. MHPO poduction rase in the IPMB wr blocked by pretreatment with 6hydtoaydop.nttne and wY increased by pretsealmeal widttseserpiwe. Towell, l. F. and Envis, V. G. Srafn ResemcA 209A76-1111, 19t1. OrAer saff.rf: U. S. Public lkahh Service and the University of Colorado National Center for Alcohol Research. From the School of Pharmacy. Universlty of Cokuado, Boulder. EVH>ENCE FOR A NONCHOLINERGIC NICOTINE RECEPTOR ON Hl/MAN PHAG(X'YTIC Lf(IKOCYTES Rah.+ligurf hrmhn6 studies show thN kukocy/es contain a numher o(peptide and Imirnrmc recepuws, such as those fnr chemrKactic peptides and a.etykhirlrne. In this ANALYSIS OF REGIONAI. VARIATIONS IN THE AFFINITIES OF MUSCARINIC AGONISTS IN THE RAT BRAIN The specific binding of radiolabekd antagonists has been used to characteriae the nwacrinie acetykholrne receptor in brain. For the study reported hue, the dfistitiea of 63 62 1

awscarinic agents in Ixains from mak Spra6ucDawky rats were determined by dircct and indirect assays of bindin6lo the receptor. Resuhs showed Ihallhe twain stem of the rnl has a higher affinity toward muscarinic a fionisls than dius the forebrain. Recepor occufanry curves of both re6ums of the braia deviate fnxn simple mass-actNrn bind- ins. The charsctcristics of the binding in each region ne compatible with the :Ristence of two nnn-interactin` binding sites, and we not attributable to desensitiza inn or to negatively cooperative binding within a small oligomer: however, the prn.-.ibihly of large ofi6ornen remains lo be eschded.'iUe agonist binding data were anal yzed by a linew transk.matian of Scatchard-like i~hibilion eurves of the binding of IIMe antago- .ist 1'N lyuin.clidinyl benzilate. Such s..lysis, bded on model of two subprpulations of rocepton in each area, shows the wbpap.laNwe of the brain atem and the forehrai. to be distinct. Brain slens: 441L of n:oeplors possess high affinity with di::sociNioa cauNant for c.rbrhol, K, - 2.6 a 10 • M. dissociation constant of low-affinNy recepux, K, - 2, ) a 10 • M; forebrai.: 4111 No affi.ity, K, - 2.1 a 10 ' M, K, - 1 71 10 • M. The data wuest that wbole Msia oorNaina r knl thrn m.r+r n wsc arinic teceptors which can be distinpishod ow t/s bosis of thek affiaties for .go.ists. FJlis 1. and floss. W. Sraiw Rrs.a.rA 19) 1t9-19l, 1950. Oder s.wp.rt: National (wstitutes of Health. From the Center for Brain Reserc#, Uaivessity of Rochester School of Medicine and Desristry, Rochester. NY (`OMPt`TiT1VE IN7FJtACTION (>F GA1IAMINE WITH MULTIPLE MUSCARINIC RkA'EPTORS Ncunl membrsnes frorn the brsins of male Spra=ue•Dawky r.u were prepared and used for binding studies with fidlamine and earbachol 1s the paper presented here, i1 is shown that Rallamine. a nicotieie anta fionist with aeMinwse.riaic pdency in several systems. interacts competitively with the trilialed ligand /'H/qui.uclidinyl benzilate (QNB) at the muscarinic recepor. TUe occtperrcy ctrves derived from these studies suggest that Rallamine has widely varying dfinilies for differeM subpoQulalions of wwscarirwc receptors. a finding which selu pNami.e aput from classical muscannic antagonists srch as ssropine and QNB. The greatest difference is dfinities far `al- Iamiee occurred in the brain sttm, whe.re the dau eould be satisfactorily fitted too two- sile wwdel, wilh 77'i o(the recepwrs havint high afBnily (K, - 23 nM) and 2196 fow affinity (93 pM). Furthet, these affinities displayed rank ordercortdning wah thnse of crb.chol (an agoaist), although Rallamine has oot, so far, displayed agonist (or partial asowist) activity. The fiadin fi thu antagonists as well as a fioeists can display multiple affinities for nwsc.rinie receptors suggests that there are fundamental differences .monfi subpr+prlarions of these receptors. Ellis. I and Noss. W. • siocArwricaJ PAo.w.acoloay 71(3) 87)-576, 1982. (hA.r trryws: Nauonal InsrNutes of 1k.hh. Fn.m the 1'enter ftr Brain RcseanA, llniversily of Rochester Schonl uf Medicine and (knlutry. RrKhesice. NY I V. Pharmacology and Biochemistry AN ISOLATED PERFUSED DOG-LUNG PREPARATION FOR T11E STUDY OF CYCI.IC GMP METABOLISM: EFEECfS OF SODIUM NITROPRUSSIDE AND OXYGEN Irn rocent yesrs, considerable interest has developed in the role that cyclic GMP may play in pulmonary function. a.rtly becwsc dte lung has a high Ruaoylre cyclse anivity comP.red with other tissues.nd p.rtly becwse the IunR is it drsect contact with variow environmental pollutanls, many of which.re tnowa to precipitate the forna- 1ioe of osy fie. bae radicals and d.me fie the Iwg. For the study prcsedod hete, the iMac1, isolated perfused dog bnj was evaluued s a nmdd for studies dioeled at defining the ndc of osdative modulrionof MMwrgeyelieGMP rnetabolism iw pulnmwry hrclion. Sodium nia..prvs.ide aided to the oerf.sios blood itae.scd the cyclic GMP cawtent of hroR over Iour-fold i.n do.e-depe.dettt owMer. Although sodium .iyv prw.ide .dmiaistntion caused changes is hrK vaacrlar resistance. these oarrred indepe.Aently of the changes in cyclic GMP. Ves+tilation of hrp with a high oxygen gas aawe containing 95% 0„ 3% CO, acwely increased the cyclic GMP coaent of b.js.tter 13 minutes /zor. 1.3 *0.06 to 3.4 10. 12 prool cyclic GMP/atR prolei.. Cyclic GMP levels returwed toward coard durinR eowlitwed ventilrion with the high oxygen concentration. The oayRe.-ind.cod ekvatiw of hwR cyclic GMP co.ten wr w1 accompanied by ch.nRes in king vascular resistance. The,rest.lts iadicMe tW the iaotred perfined Irg fi.my be useful i. studiea of cyclic GMP, tissue oaidrio..nd pMaw.ry fr.ctio.. trorsAln, l. df. and Muon. M. B. E.ropr.n lorwef of PAmwsscoloty 7i:1t7-19),19R2. OrAer ssff.rt: Aatericr Lrsag Associrios. Prow t!e ProRr.nr in PArsn.oolo fiy and PhysioloRy. Nordresfler.Ohio Uwivaside. College of Mediciee, Rootslow.. ' INVOLVEMENT OF SULF7IYIMYL GROUPS IN 7HE OXIDATIVE MODULATION OF PARTICULATE LUNG GUANYLATE CYCLASE BY NITRIC OXIDE AND N-METNYL-N-NITRO-NITROS(X;UANIDINE This reporl describes the activation of p.rticdne rat lung Ruanvlate cyclaae by nitric oxide aed N-methyl-N-.iw-N-nitrosotu.nidine (MNNG) and suggests the i.- volvement of sulfhydryl `roaps in the tlctivatioa process. In this swdy, p.rticulre gu.nylne cyclase from rat lusfi was activated by witric oxide or MNNG in a do.e- dependeM manner that was affected by dirhiothn: itol. Although low eoncentralions of ilric oxide or MNNO produced ner maximum activMion, excessive amounts de- creased the particulate enzyme activity. Nitric q.ide-Mimulred Rranylae cyclne activity decayed durin fi a 60-minWe preinc.batiom period r 3TC, but dyd wd decay r 24' or 4'. DithiotMeitol enhanced the decay of nitric oside-alinmlated en:yme at all tempa.wres by pmcntiatinR the revcnal of nitric oxide activation. Folbwin fi the reversd of nitric oxide activation at 24' by dithiotMeitol, tlhe p.niculue en:yme could 64 ' 1 65

.bc rcactivated by a seclmd expxure to nitric oside. PreimuluNinn of hasal Paniculate Ruanytate cyclase activity at )7' resulKd in the loss af enzyme respmslveness lu al 1ovalnin hy nNr.c oside ot MNN<i This loss of respMsiveness that was prevented by the Ihwd aMMraidaMs was ptentialed by the tMol oadan/s, diamide or osidored 61u- lathwmc. Also, the mhiMwry effects of the Ihio) otidaMs on enzyme respinsiveness to acuvatnon by MNNG were prevented by dithiolMeilol. These results suggest that the aruvatwoe of partoculate guanylate cyctase by ailzic oaide or MNNG involves the osdatwn of key enzyme suUhydryl popa. AraraAfer. J. M. ainl'hnwk-e/ PAarwrwolo4y ]I(7):1239-1244.19t2. Fnwn the PrlKram in PharmacoloRy. Nordteaslera Ohio Universities Cdk4e of Medi- eine, Ruwsarwn. SEPARATION AND DETECTK)N OF UPOPROTEINS IN HUMAN SERUM BY USF. OF S12E EXCI.USION LIQUID CHROMATOGRAPHY: A PRELIMINARY REPORT This mnhednk.gical puofier pese+ws a relativcly mild and quite rspid procedure for separatin6 serum lipopnocins far individual eolkction. In a firsl step, hman serum compwrcnts, including lopiprvlci», can be rapidly separMed by sizetscUision "hi6h- prrfomunce" liquid chromatography. Then. liprproteies in fractiaa of t're eluNe can be quantaalod by conventional chemical and enzymatic methods. Alternalively, if liproproaeins in she serum we selectively prestaired with difonnazan dye, the cdumn effluent can be munitored spectmphotanetrictlty at 5!0 am, so that oal) the hpopro- Icin components uf serum we detected. Samples of purified low-density foppqnuleins, so stained and r+alyzsd, pnvidc p:ak•area values that we proportionallo their cuncen- Iratron as evaluated by chemical methods. With this kchniyue. the various Iipiprotein classes can be quickly separated and their concentration estimaed. Overall, these techmilues aeem so have potential (or development into analytical and clinical p\Kedures. aarbrr, n. L. er ar. Cliwirof CAewisrry 27(12):2052-205f1,19t1. Other r.Maarf: American Cancer Society. RubeA A. Welch Foundation and a North Te.as State University FacuNy Reserch Grant. f.om the Departmcnts of Bicftical Sciences and Chemistry. North Tesas State Univenily, Denton. and the [kpartrneM of Biochemistry. Tesas Colkee of Oileo- pNhK Medicine. Ft. Worth. A NEW SUBCUTANI:OUSLY-IMPLANTABLE RESERVOIR FOR St/STA1N1:1) RIJ.EASE OF NIC'OT1NE IN THE RAT Nwo.rsne has bec/.mc a wodely studied dnrg for its phannacoM6rcal and w.icolo• R« .1 r16/ ts Iw trrv..us .n~mal sludKS, IurKd dminblratNM mtlh~dl f~w m/'/Nlne in. IwIrJ P.remcial m/cl tn.n. afmmistrNwn hy mhalauon ld n6an ne sm~Ae, s~~lubd~- zilion in drinking waler, and injectian in single do.es or chronically via Alzet mini- panp into the ventricles of the brain, anwMng others. This paper now reprcls the successful developmenl of a subcutanerwsly-implanlabk reservoir Wr the sustained release of nicotine. The device, dubbed INR for Impl.ntabk Nicotinc Reservoir, is a small glass cup sealed wrth Silaslic* pdymer. It releases 3.4 mg of nicotine per 24 hawn. When implanted into moderatcly-sized femak Spra6ue-Dawky rats it podaces bhrid nicotinc levels of 400-500 ns/m1 which tenuie relatively staWe over M kasl 18 days. INRs are nrnwosic, reproducibk. inesp:nsive, and adaptabk for pharnucol*- eal and Ioaicoh+gical studies in rMS and oMet small animals. Erickson, C. K. er al. PAa.a.acoloty S/ocJr<erArry f Sr/bvior 17:1t7-1t5,19e2. FnMe Drug Dynamics Instilute, Culkge of Pharmacy. The University of Teaas, As1in. DETERMINATION OF THE PRIMARY METABOLITE OF CENTRAL NERVOUS SYSTEM NOREPINEPHRINL, )-METHOXY-4-HYDROXY- PIIENETHYLENFGLYCOL, IN MOUSE BRAIN AND BRAIN PERFUSATE BY HKiH•PERFLNtMANCE L.IQUID CHROMATOGRAPHY WITH EIECI'ROCIIEMICAL DETECTION The rab of p.oduction of 7-mNhoty-4-hydroayphenethykne`Iycol (MHPOh the principal rnnabolite of norepinepMia in mamalian braia, has been p^opased to be a. indicalor of the rate of sNUepinephrine ur.over. For this teaso., new methods of MH?'Q isolation and detectian we important i.itrtherinR man's knowledge o(the tole of nc.epinephrine in normal and dysfwtctioaal m.nulila bnin. ln the present method- olo6y p.pcr, assays we described for the detersni.atiott of picomok levels of MHPO is nrouse brain and in the petiusae of an intact ttsorse brain. High-performance liqaid clwnnato6raphy with eloctrochemied deteaion yielded a MHPC, detection limil of 0.37 pnd. This technique offen a sensitive and ineaprnsive allernative to ps clromr tography with mass specvometry, and can be used in tonjwrc/ion with braiA catocho- lannae deleflmnanoas. i Towed, l. F. and Erwin. V. C. Jawwof uf CAronwtoarqp/iy 223:295-303. 1951. OtArr z.ayp..t: U. S. Public Health Service. From the School of Pharmacy. University of eolotado. Boulder. PRIMARY STRUCTURE OF THE SICNAL PEPTIDE OF TROPOELASTIN b Elastin is a majnr slroctural compmenl of carrcctive tissues. Its sgiubk pecur- aar, tmplelastin. is e.tractabk in organic solvents 1ed possesses an estensive clusler- intot mmpolar amino acid residues in the immediate NH,•lernwnl region. Since many poleins secreted from eu\arywic cells are initially synthesized with signal p:pides, 1ropoelaslin was esamined to see if it did in facl contain a signal p:pide. Leader .oqnences, ur si6nal pcpndes, we relatively shon hydrophobic NH, terminal eaten- ~ I 67

sions, whk h are tM)u6ht to play a rok in veclorial transpat of the nascenl pslyp:ptide. Thcn:ftre. the possibbiny that the imtial lnipulastin tsanslalnN+ puducl pnsesses a shnn siRnal Pcpide was esanuned in a cellfree translation system. In this study, tdal RNA, isulated from aonae of 1-d.yofd chicks, was translated in an mRNA-dcpendent relKwhK yte lysale uamlalNM assay. The translation products were then /mmunnpreci- ptated and subjected lo automated radiosequencieg. ComQrisoauf the NH; tennind sequence of tropoclastin b synthesized in the cell-free system versus Uut synthesized in orkan cuhurc dcmunstraled the ptseace of a silind pepide 24 amino acid residues in length. The signal peptide sequence is n tollows: Me1-Ar6-GM-Ala-Ala-Ala-Pro- Leu d.cu-PtaGly- V al-1 ~eu-Leu-l.e.-Phe-Ser•Be-Ltr-Pm-Ala-Ser-Gln-Gn. The pe- pwwlerance o( hydrupfwibic amino acia tdidres as well as the p>tr residues adjacenl to Ihe initrator melhionine and the earboayl lersnini kwnd in the signal p:ptide is similar to that reported for dher aecreled poleiws. Krr, S. R. and Fosrer, !. A. The lovnd afslolotkd CAnwlsery 236(12):3416-3919, 19R1. OtAer sspla..t: Nriond Institutes of Hedth. From the 1)epartment of Biochemistry. University of Georgia. Athens. ABSENCE OF SEASONAL VARIATION IN ANTIPYRINE METABOUSM Aryl hydracarbow hydrosylnc (AHH), the wro.ooaygenase syaem(s) that me- Ubdires henro(a)pynrne to fluorescent phenols, has been shown 1o eahibit a atronS seasowal variatan The ra vrro metabolism of rMipyrine, which is also catalyzed by wrictva>fnd cytocMome P-130dependeM snonooaygesases. has been reported lo be correlated with AI111 inducrblily in human lymphocytes In the audy ptsented here, • an attempt was made to dctermtne whether rwipyrsne melabolism, like Allll mcNabo- 1ism, daws seasonal chantes To do dus, rwtPyrure half bfe (t l/ 2) was m:asured in 10 a<rnsmulten and eight smuken in the summer and the wimer, the two time s of the yer dW catespond to the high and low peaks of inducibk AHH activity as measured is lymphocytes. Results showed UW 1he rttrm anupytine 11/2 determined in all 11 sub- jects in sunmer was almost identicd to 60 found in winter (s!SEN - 10.90 2065 and 10.96 t0.7t hr). AHH activity id c.Ntred hum.u lymplwc)tes from the •oesmokinS subjects was delermiaed ia eo.uol and S-mnhykholantluene-indaced cells to oblain indreibilNy ratios of 4.2±0.56 (SEM) in the summer rd 1.130.1• (SEM) in the wiwNer. These tesuhs indicate 1ha11he seasonal variation in AHH induci- bility in buman lymphocytes is nd reflected by a corresponding seasonal variation in antipyrine metaboiism M vino. Pairn, B, Ward, E.. Steenland, K., Bolrawska, W.. Gtasne.. T.. Chans, R. L., Wood, A. W., and Conney. A. H. Cfinkal rhonwero/osy A T/kr.qewici l1(2):I{I-1S0, 1952. UrA.r, raris..t: National Cancer lautitute. Fnwn the 1)epartments of Mokculr Biololly and Esperimenld Therapeutics. Roswell Park Memexral Insistute, Hu//aWi, and the [kpartrnenl of Biochemistry and Drug MctaMdo.m. Ibllnunn IaRiKhc Inc , Nulky, Nl. ENZYMATIC PROPERTIES OF HUMAN GI-UCURONYLT:2ANSFERASE AND A SENSITIVE METHOD FUR ITS ASSAY IN A STABLE 8 LYMPIKICYTE CELL UNE UDP-6lucuronyltransferases are membrane-bound enzymes located chie/ly in the endoplasmic reticulum of cells and arc responsible for the conjuption of endornous and aenubidic substances contsining hydiosyl, amaro, thiol, or erboaylie functional groups. Although 1he.e is much interest in huma thKUmnyNra.skrases, eauensive studies af them have been hampered by difficulties associated with procurement of sufficient quantities of fresh human cells. This repart, however, dennoastraes that a prolific stable cell line, such as SNIW6, capable of yielding billions of cells ecald be used for such studies. IlomogenMes of SNKI06 cells have been studied. A sensitive assay procedure for lymphocyte ghrcwo.yltranskrase was developed utilizing sadio- aclive leaouerone as the acceptor substance and TLC for seprnioa of the melabdNe. The mctlwd is capable of desectiwll pcomolr quantities of the product. The enzyme activity ednbited a hroad P11 oplimwe, and was subject to aclivMioe by the deterSaM l.ubml W X and Mn •• ions. The rtivity confonned 1o the Michaelis-Meaen kiwaica giving apparent K. values of 0.8 adN and 11 µM, for UOPGA and testoaeso.e, seapectively..-MeWyhrmbenaerone, a-.apMhol and P-niuophenol behaved as oa.- p:titive inhibitors of tessoaaoae glucwonidrion. The sesuMs preseMOd in this ppa indicate that sae melhod could be rsed for Se.e/ic studies of humr lymphocyse ~ Shw<wonykrwfer..e, and thN 1he enzyme is of eo.seqrence in detoaicrion of eto- - e.ow aa well r endornous subslraW. Li, H. C.. Porter, N. and Genwn, T. Enrywr 21:5l4S,19i2. Ori.r a.ff.rf: U. S. Public Health Service a.d thc New York State Oeprteaea of HeaNh. From dte Deprtmer of Eaperime.ul 7leapeaia a.d (kace Cancer Drug Center. RwweB Prt Memorial lawilule. Buffalo. SUBSTRATE SPECIFICI7Y OF HUMAN UOP~',LUCURONYLTRANSFFRASE IN CULTURED LYMPHOCYTES Human lymphocytes possess `hrcuweyMraesferare activity with a btod rase of substrate specificity. For the papcr pescntod here, catalytic properties of lymphocyte ghicuronykransferase were pudied, and aclivily for the following substances was doerweenied: testostemee, estaadiol. PLenolpMhakM. u-napMhol.1-mefthumbelli- feronc and o-nitsopAend. As has bee. nded before. e-n.pFwhd is regarded as a model aSlycone for dse form of khicwonyNransferse which erdyses ghrcuronidrion of the less bulky nonakroidal srbstratcs, and leswsterowe as that for 1he steroidal qlycones. The results show that humr lymphocytes possess tArcuronyhransferase activity for both types of allycone. Within IimilNiaa of work with crude hamokennes sad 60 miautu incubatioro. no difference could be seee in apparent K_ values of UUPaA 68 69

when the various afttycone substrates were tested. Funhernw+re, compctiti.c inhibition of tesuntcrvne UI)P-6lucuninyltransferase was observed when 1 mc:hylumbelli- fermne, a naphtMol and p mtnrhenu/ were used as the inhiAiurs. These data demun- slrate the simttarity of the entyme(s) accepting the varxws substrates. l.i. II. C.. Pbrser. N., Ik.lmes. d., and Gessnrr. T. Xrnof.ioaire 11(10) 647-651, 1991. OOther aqr..r: New York State Deparuneat of Health. Fmm the 1)efwtment of Esperimetatal Tltaapeutics and Grace Cancer tku` Center. Roswell Park Memorial Insulwe. Buffalo. STIIDHFS ON T11E IWPOSITION AND DISTRIBIITION OF CATFCINH. tR(MWfKH-E CIGARETTE SMOKE IN BCFI/CUM MICE The studies reported here were perforsned to delermane the depos/tan, dlstrsbr- tion, and ckarance of catechol in cipreue smoke using defined smoke e.postre condew,ns and a weY-charscterinod mwse strai. (BC3F1/Ctnn). 'flte ptesence of 1'll/catechol iw the smoke was verified by silica gel cMomalography, hith perfarsn- ance liqusd ctr.wnatugnphy. and ps cMorn"raphylnuss sqeclrornelry. Mice were esposed to 10%(v/v)2R 1 cigarette smoke on dte Wahon Ilori:onMal Smoking Machine under standard condnions o( 33 rrd puff volrme, 2 seelpuff. 10 puffslcigardte. The deposition and distribution of inhakd catechol were determined in all internal tissues, rrine. and feces 1)ata showed that clearance was occwrinS during the I0-min smoke esposure period. Immedutely after esposure. over 30% of the radioactivity was found in the bbnd, with 10M found in the brtg, and approsirn.lcly 12% in the respiratory tract. Over 91% of the inhaled radioactivity was found i. the trine 120 min dter espoarre. Less than 0.3% of the 1ou1 dose was found in the lung M this time. 1n sunraary. these determinations show dut eMecbol in smoke is rapidly absorbed, redistributed and escreled frorn mice espoaed to whok cigarette smoke. Ilwang, K. K., Sonko. 0.. Dansie, D. R., Kouri, R. E., and Henry. C. 1. (Mk.obio- foRicd Associates) Toikoloty drd Applied rAarw.acololy 6/(3):I0S-111, 1992. FFrom the Department of Eaperimentd Oncology. Microbiological Associates, Be- drcsda, MD. EFFECTS OF NICOTINE ON UTERINE BLOOD FI.OW AND INTf:AU7ERINE OXYGEN TENSION IN TNE RAT Since i1 was known that normal concepus developtnent nuquires optimal levels of osygen, tt seemrd reasnnaf.k so determine whether nicdine-inducett reduction in utettne M..d IIM.w coraiMnotanuly decreaser the crwrcenMratMm of oayf[n wNhin the utrnar tumrw In ihc uuJy prc.eMCd heee. N wsr seen thal suhcuuneois intection of 1 nicotine (0.5 4r 5 m6/ks body w1) resulted in a marked and prolonged reduction in ulerinc blood flow and inuaulerine osygen tension in pseudnpre6naM rats (Day 4). By 10 minutes after nicotine administration (5 mglkgl uterine perfusion was reduced by •0'R, remained suppressed for 90 minutes and eeswwtd to the pre-treatnsenN kvel by 120 mirnwes. Rats receiving the 0.5 rrtg nicotine/kg also showed a narlced roduction in werine blood flow, although the response was slower in onset and IonSw in duration. Nicotine (3 ma/kR). also resuNed in a swt.ined deerease it aMrawetiae oayeen tension ftvm a conlrol value of 18.9 i 3.61o 22.2 ! 2.6 rmn11R a45-60 minaes ard 21.7 s 1.5 mmllg n (0-90 minutes. The freqrency, and ampl«rde of Ottclu.tiows in inltarterine osygen tension were still reduced by 90 rninwes after uealmad. Tl.aefotee , tite resrMs of dus study indicate dW nKOUne, in ahsounls srffic/eat to suppess embryonrc powtA, reduces urcrine blood flow antl producesa marked and srstained decrease ia osygcrt tension within 1he uterine lumen. The time-cotrse of the reduction in iMr.wer- ine o.y6en availability parallels tlul of the nicdine-induced decrease in uterine blood Oow. Ilarnmer, R. E., (~ildm.r, II. rd AIitrAtll. !. A. lor.wa/ oJRep.odrrriow oid FerriNry 63:163-16R, 19!{1. From the DeputmeMS of Anatomy and Pharmacology. Wayne State tlniversiry School of Medicine, Detrvit. SYNTHFSIS OF NON-K-REGION ORTNO-QIIINONES OF POLYCYCUC AROMATIC NYDROCARBONS FROM CYCLIC KETONES h this mdhodoloRical paper, uo.-K-teRioe o4trinones of Po/ycyclic arornalic Mydtvcrbons are pepred easily by Aeating a sottrtiow of a letrahydrodiol with 12 eaiuiv. 2.3-dichlotv--S.6dicyano-l.l-beazoqri.o.e (DOQ) in dioaane (re0ra. 24 Irs). DDQ not only osidixes dte 1,2-dio/ moiety a-0ikaone bu1 also Mtroduca the ok6ak duubk bond of the o-qtinone. When this one-slep convenion was first disoov- erod. dse surprisin6ly f.cik generation of dte o4fuinone was attributed w the ease of double bond forsnation u the bay-tegion as described for several tetrahydro arcnes. Nowever, present results indicate thr the reaction is wot restricted lo the syndrcsis of non-bay-teSion o-qukeones ba is also applicable a the prePaaion of b.y-region o- quiwwnes. llws, dtis method is both simpler and anae general than qte nsonMly published synthesis of non-K-region o-quinones. Overall, the synthetic approach pre- sented here constitutes a new nethcd for the prepsrNion of non-K-region oquinones dw seems to be 6eeer.lly applicable. Pfrt. K. l, and Oescli, F. TnruhcJ.on l.etrea 23(2):163-166, 1982. From the Department of Toaiarkgy. Institute of Pharmacology. University of Mainz. M ii.z, Federal Republic of (;ernmany. 71 711

IMPROVEI1 SYNTIIFSIS OF ('-1•TRANS-9•lO-DIHYDROXY-9-N) f)II IYDRO- BI-.NZC)jul PYRENE ANI) OF (!)-TRANS-1-2,-DIIIYDROXY-1•2-DIIiYDRO- Dlltl•Nl1a.A/ANTI IRAI-ENE Non-K region dihydnodiuls o( pslycyclic aromstic hydrocarbons (PAH) play an Mnpxtant ruk in the metabolism of PAH. They we precursors of dihydndtul ep)s- ides• some of which are considered uMimate carcinotenic metabolites of P\11. While the non K regittt+ dthydrodiols with the okMic bond in (he bay region have beea iNensively studicd, the biological tolt d drc wots-K-resion dihydroditas of ben• ro(a Ipyrcne aed dibenz/a.Al anthnrceae (6b atad 17b), where the ukfiaic bond is not of the bay regiun, is kss well know.. la Ihe present study. improved synthetic pathways for hb nnd 13b were devised tha/ ttwMod in Ihe schemes seen below: a sis-slep synthesis of 6b (Scheme /) and ei6ht-uep synthesis of I lb (Scheme 1I). 1L"tit k_ I l r "v Cj ui LY ® .~ ~ u>_ ~ (J • ~•w r r r.. ~ r s... ~ n • '"`~ ( ~" Z DC7 • Go 0 •~ n~ Z 1J ~-Y 1 r- .' ~. . ll ~~ a 0 Since the link interest in the dohydrvdiols 6b and ()b items panly from the fact that their syntheses descnbed in the hlerature are more difficuh than those of the better tuudtod isomers. n is hoped that these ncw improved syntheses may help in furthering study of the biological roks of bb and I3b. OeuA. F. et d. The Jorrwal aJOrtoRlc CIKmnlstry 47:561,371, 19s2. From the Inuitute of Pharmaadoty. Section of Biochemical Pharmacok+gy. Uoiver- sily tt( Mai.z, Mainz, Federal Republic of Gerasawy. K-REGION TRANS-DIHYDRODIOlS OF POLYCYCLIC ARENES; AN F}-}ICIErfT AND CONVFNIFXf PREPARATION FROM O-QUINONES OR 0 DIPHENOLS BY REDUCTION WITH SODIUM BOROHYDRIDE IN THE PRFSENCE OF OXYGEN K-regiow srunz-dihydtodiols we importsnl metabolites of polycyclic aromalic hydrocrbtru. They we produced by epsside hydrdse-mediated hydralan of the primarily formed reee osides. The K-region Laiu-dihydrodiols we usually synthe- siztd by steteosekctive reduction of the easespondinS o-quinones with aMnpka metal hydr.des such as hUiutn duminum hydride. When fust attempts were made to use sodium or potassium borohydndc in methaail or ethanol (or the reduction of K•re6ion o qutnones, they were not very successful. Also, the use of dilferent reaction limes and temperatures. diffcrent nlHn of the rcactrNs, and different solvents did not improve .. the results substanually. However, it was finally found that polycychc o-quinones can be n.nveni!nlly reduced to the carsesp>n<lin6 nonz-drhydrodiols by carrying oul Ihe reduclion with burohydrxks in the presence of osy6en. The nde nQ osy`en is the reduction may be rationalized by a mechanism presented in this paper. As a resall of the reduction-osidatwa cycle thu takes place whee osy6eo is availabk, pure trans- dihydrodinl accumulates so thal it can be isolated in high yield. This new mahod of te+ducins K-regrm oyuinones ;and dso the corresponding odiphenols and their ace• tresl derived (rom polycyclic arenes represents a significant improvement of the synlhesis of K-re6iort nansdihydtodiols. Plrt, K. L. rd OeuA. F. SYNT1/ESIS : lnurnodowd Jownal of Medbdi !n Sywdieric Orlonk CAnwistry No. 6:IS9-162, )une 1912. From the Instilute of Pharmacology. Section ow Biacherwical Pharmacology. Utuva- . .ity of Mainz. Maiaz, Federal ReptElic of Getsn..r. I ANALYSIS OF NICOTINE AND COTININE IN TISSUES BY CAPILLARY GAS CHROMATOGRAPHY AND GAS CHROMATOGRAPHY -MASS SP6(TROMETRY 1s this methodological paper. Ihe developarcat and applicMion of inethods to qaaditate low levels of nicotine and cotinine is tissue samples we described. Analyses were performed by capillary eohwm gas chromatography with a specific nitrogen- phosphorus detector and by gas clront.toRraphy-mass sp«aometry. With close wrc- twd analogs fur internal strdrds. hieh auottitative accuracy and pecisiots wee de.rowstrned f« the range of s-1000 ag pet e of tiswe. The sensitivity limit was 2-3 aS/R for both compo.rd.. lle .uia advwaRe of Ihese techtsiques compared a previously published methods is itsctened selecti.ily; the other meqwds were devel- oped 1« analysis of biological Arrids atd ate nol readily adaptable to tnae cowiples biological mwiceu such n tissue homo6ert.les. With the newly deveMoped teciniquet, it was possible to perform a pbarnncokiwdic 1Ntdy of wieoline and cotidne is mouse iiver following a single iatraperiloaed ittjection of nicoliae. 'lbomp.ora,1. A.. Ho, M-S. and reu.sM. O. R. Ja.rwal o(CArow.ototrapAy 231:s3-62. 1912.. From the School of Phrm.colopr. Uaiversity of Colorado. Boulder. RELATIONSHIPS BETWEEN CHEMICAL STRUCTURE AND CHOLINERGIC ACTIVITIES OF FURMETHIDES AND POTENTIATION OF THESE ACTIVITIES BY PHYSOSTIGMINE 'flThe fursnethide compowtds-furiurykimethylatnrnoniwn ((unnethide: FT), 3- snelhylfunnethide (5-MFT). S-hydnosymethyHtwsnethide (5-HMFT) and 3<hloeo- nxthylfurmcthide (St1MFi)-are pnten/ and suble ew.catink agents which we being used in pharmaco{otical investigations whne nwscrirtic selectivity is required. For die studies repxted hen:. the cM>tinergk activities of Ihe /oar furmedtides wese compned on a superlused guinea pig ikum peprarion. Results showed Mat JI four components were less putent but equally as active as acetykhuline (ACh). The EDSOs 72 73'

of Ihese awnrywnds dcrnasc in the Gdlowin` iwJcr: S C I MFT > S• FIMFT > FT > S- MFT The ckclnon densities aniund the (ururusygcn Mo„ns of these cumpwnds decrease in the same unkr A careful analysis af the effects o( these a6cnts twr the =uenca pig okum revealed that the furmethrdes display two separate actions ( I1 duect inleractNwr at the muscanna reccpor to ebcM a rcsponse, and (2) release of ACh from the presynaMrc nerve terminal In the presence of physoui6mine, dose respor se curves of these compounds were shifted to the left. There was approaimately a two- foW shift in the EI)SO values (or the (unnnhide. Like aracarine, these cnmpounds were not hydrolyzed by cholinestetase (("hE) At kasl p.rt of the noted (urmethrdes re.pmse was due to release of ACIt, because the respo~ses were increased by CAE inhibition. Chaturvcdi, A. K, Rowell. P. P., and Soxry, e. V. R. P1io.n.ocofaeicof ResrorcA ComwwwicaNowa 13(9):E29-IhS, 19t1. OtArr s.ipla.rr: National Institute of Child Ikahh and Iluman [kvekpmr.v Fnan the (kfwtmem of Pfiarmacok>ty, Vanderbih UniversNy School of Medicine. Nashville. 7N. (fiANGFS IN MICROSOMAI. MEMBRANE MICROVISCOSITY AND PFN)SI'/R/1.IPID Mt:THYt.1RANSFTRASES DURING RAT LIVER RF:(iENERA f 1(NN Re6enersung liver is a good esample of a controlkd growing tissue Durin6liver amwth, newly symhcsired mnkcuks of drvg metabolizmg enrymes like cykschnxne P450 are incorporated into the fiP«f marns of endoplasmic rcticulum IER) There is a rrrur/w+ln6ical associuinn of nbnsnmal PnMnn synthesis with ER membranes Recent research has shown that decreased membrane microviscosity facilitates incorpuration of the enzyme protein molecules into the yhnspfwrlip+d nuitrix of ER. The micronscos- ity of membranes is ahered by. 111 changed ratios of phospMrlipid to cholestenol.(2) wtsanwalwm o(fany acds in pMrspMdipds, rd (7) melhylrion o(phosl+Aulilwds. In a related study. miaoviscosity and S-adcnosyl-L-methionine (SAM) medialcd methyla- tinn of ptrrsplsatidykthanolamine 1o plasphatidyl-N-meU+ykth.nolunine (PME) and plwnrhatidykholine /PC) were measured using micrwaeal membranes of rrsencraf- ing rat liven at 6-96 hn after partial hepatectomy. In the methytned phosphidipids, the I+ropntion of PME increased by 3-99i at I µM SAM, and, a 2U0 µM SAM. the proportion of PC docreased by about 5-10'Jk at 12-24 hn. Two phase transitions were observed with micros.Mnal membranes between 20 and 40'C. In synthetic lipnrxnes containing PE. PME and PC. rnicroviscosity decreased when the proQrxtion of PME increased or the prpontion of PC decreased. There(ore, alterations in phospholipid methyltransferases and consequent changes in membrane phnsPhe>fipid methylation may corwitxrte to increased membrane fluidity during cell pruli(eralion and incrmpira- tion of drug metabolirinR earymes into ER. Jaiswal. R K, Sotrry, d. V. R. and Iandm. E. 1. PAorn.wohoRy 21:335-365, 19t11 (hher support: U S Public Healih Service. Fmrn the (srputmem o/ eharmacs.kogy. Vanderbilt 1)nivenity ScMwd 4 Medicine. Nashvilk. I N F:NIIAN('I:MF.NT 01: TIIE RFSPONSIVENESS OF TNE RAT DIAPI/RAGM BY L-MITHIONINE AND P/IOSPIKH.IPID METHYLATION AND THEIR RELATIONSHIP T() AGING The contractions of hemidiapMa`ms of Fischer 344 rut of age 2 to 33 moNM were measured after electrical stimulation of the plrenic nerve or the muscle in vitro. Earlier work has shown that S-adcnosyl-L-melhio.ine (SAM)-mediated methylatio. of membrane phosPholipids incrases membrasx fluidity and respo.siveness of the asusck, and that an increase in the inuacellular kveh of S-adeuotyl-L-haaocystcine (SAN) inhibits pbospholipid metd ylalion. In the study presetNed Me, the inlncellular kvels of SAM were increased by incubring the hemidiapMaRru in L-medtionine. L- hornocysleine thiulactune, adenosine and erythro-9-(2-hydroty-3-wonyl) adenine. an WJibNrw of adennsine dearniwase. The following tesuhs were obtained: (1) Microso- mes 'rom hemidiaPMasm conained tpid mnhyhnmkrases. (2) L- nathiowine increased muscle leanwiwdcve~ekctrical sfiowluiowof the muscle or the nerve. This increase in the tension is dependent siPo. the concentration of L- meMhir*tine. (3) labckd methyl ptwps were incorporafod bom labekd L-nxthiowine ido phos(iholipids of rhe hemidiarslra`m. (1) Ptesewce of dewti.e, L-bomocysteine thiolactone and erytMo-9-(2-hydrony-3-nonyl).denine i.hibiled the effect of L- methiordne fo increase musck tension and incorpaalion of methyl poups into phos- qholiqids. (5) Muscle tension developed by electrical slinwlaiow of the plsn nic suxve or the muscle decreased with increasing age,.nd (6)'T1te met6ionine effect was erralic or insignificant in hemidi.plraRnu of old rMs (> 13 momhs). These observations indicate that increasins the cellular for.aatioa of SAM increases phosplalipid methyht- tion and the contraction heights of the hemidiaplra6m s+pw electrical stinrrlssion. Bah o! these e((ects we inhibiu+d by inaeasinR kvels o( SAH. They also indicate tlas SAM-malired membrane plwspholipid methytatqw sss,d fine regtrlation oI mcmbra.e Ifwdily in 1he diaphragm may becrome de(ective wiwh advanei.R aRe, and m.y contrib- swe partly b 1he functiand deficits of the diaphragm. Saury, e. V. R., Owens, L. K. and l..son, V. E. Tlk lorrwol aJPAornsocoloty owd Eaoe.iisenrd IArnwtwkr 221( 3):629-636. 1992. OrA.r saw.rr: U.S. Public Heaph Service, National Irwitwe of Agin=, and dte National lstitute of Child HeaNM wd Human Devetopmenf. From the Department of Pharsn.cobgy. VanderbiM University School of Medicine. Nasbvilk, TN. I RFd:U1.AT1()N OF ACETYLCIK)1-INE RELEASE IN THE M(N1SE CEREBRUM BY MITNIUNINE ENKEPIIALIN AND SUBSTANCE P Meshionine enkephalin (MEK) and Substance P(SP) may regulate acNykholit,e (ACh) release in the cerebrum. In the present Mtenspl 1o study this procesa, mouse cerebral slices were incubated in a mndilicd Krebs bicarbonate buffer containing (mnhyl-'N) choline chMnNk The slices were fihered, washed and vansferred so a mocrubath sn up fur superlusion. ACb release was followed by she e(llua of labeled 74 75 '

ACh and choline as a funclion (if lime. The rekase of opioid pcptides was measured by spccthc tsditMnurNUtoassays The rate of'H•ACh release increased initially for the first five min., reached a peak, and then declined capnnenlially (half 1ime, 73 mtn.). Electrical field srimulation of the slices during dte eaponenlial phase caused a si6ni- ficant increase i. ACh rckase ftw five min. Absence of esngeneous Ca" in the pcrfusrnn field depressed buth spontaneous and evoked release of ACh. Wlten the effects of kins acting enkephalin (D-aiatnkephalinarnide. DALA) aed SP on AClt release were sludied, DALA (34 nM) decreased the release of ACh (65%) as •well as Ca" uptake(111%). SP(6 x 10 'M)irrcreaaedACArekase(l3%)andincreaxlCa'• uptake (154%). These obscnations iwdicaae dtu enkephalins may esert a ncsative feedback control of A('h release from cerebral sliees by inhibiting Ca' ' uptake by the slices Similarly. SP may esen a poutive fetdback on the ACh release by enhancing the uptake of Ca' • by cerebral slices. Snrrry. A. V. R. ard Tayeb, 0. S. In: fMaw.n, B N. (ed ): Current Srarrs of Cewnnlly Acriwe Pepridrs, New York: Pertann. Press, 19112. pp. 165-172. (also i.: Ahwrces /n rAe Aioscfences: 165-172. 1962.) OuAer nNwf: U.S. Public Health Servra. Fnwn die Drp.rdrreu of Plureucology, Vr+desbiM U.ivasity Scbooi of Medicine, N..Avilk, TN. ION BEAM TRRIUM L/1BElINO OF PROTEINS AND PEPi1DES As advanoed technrque for tritirin~ prweina dtat yields high specific r.di.mctivi- ties without causing sismflcant changes in biological activities is peseMed here. In this method, a carefully controlkd panick beam eompo.ed of T,' and T,' ioas nri fast T, mvlecuks is .cctknted iew a sample target widti" a vacuum chreba. This beam method has beea taed in tritiMe ribossrckre A, poteine paecreatic elnlaae, drcrnxdy- sirr, soybean tryp.wr inhibitor. a; proteue i.hibilor, ard the peptide aldehydes kupep- tio and awlip.i.. AAer renaval of all readily e:chawgemDle tritium, tlre products were obtained in )2-2)X yields with speciBe rsdio.ctivities o( /1-d36Ci/mol. The products were carefully cA.r.cterized, show. a be chemically pwe, and to have complete biological activity. Simple Irilium hydrogerr eacb..ge.ccounts far at kast 92% of the reaclion pathway with proteins and for 100% of the reanion with the peptide alde- hydes. This metAod is practical awd economica! o. a reasonabk scak, producing high sqecific tritiurn incorpor~tioa withoul significanl ehernical decornposilion or,wctural change 1o even fragik, high molecular weight malerials. Bsuh, (1. A. n a/ (Trovis. !.l Thrla.rnoJ o/SiologKaf CAeiw.ary 2S6(2l):12217-12221, 198 1. OsMr r.ryNau..w.l lauaures of Ik.NM Fmm the Sah.rd of (1.rmuuy. (:c.wgia lasutute of Tcahm.bgy. Atluua. ISOL,ATN)N AND PROPERTIES OF HUMAN NEUTROPHIL MYELOPEROXIDASE Purificainn of human kukocyle mycktpemsidase, as described in this paper. is. simple dree-step procedure involving dialysis of agranuk estracl against low-sah brrff«, Sephadea G-7s chrorrntography, and carbosymethyloellulose chrornaaoS- raphy. The final product of this purifrcation is homogeneous when esaminod by arid pnlyacrylamide 6e1 ckctrvphaesis and sedimentationequilibriuas ultrscentrifullnio.. The mokcular weight that was delermincd hete by the latter procedure was 11>l,000. With or withnut reduction of the protein by 2-mercaploedmwl, suE.rrits were forsrlod which migrated as a single b.nd after sodium dodecyl sulfsfe Sel ekcdophoresis I tic . molecular weiaM of the appareMly identical suburtib was 59,000 with reductioa, and 12,000 without roduction. Other general properties of human kukocyte myeloperoa- idase, including amino acid camposition, amino lermiru/ sequeace analysis, and ab- wrptioa spectra, we also reported. 7Nyelopetosidase, in the presence of hydsoSen peroside and chloride ioa. and no other wbshase, auloi.activres. AAa completion of the inactivation reactioa, several osidizable anwno acids is die enzyme we modiGod, smd the abaaptioa peak a 130.m disappears. The preser.ce of substrate of the myelo- peroaidaae system(a-l-protei.aae inhibibr). or of high ca.cattr.tioaof chloride io., eompldely prdexts tlre enzyme from nMOi..cuvatio.. Matbeso., N. R.. WoaK, P. S. and Trads. J. s/ecAerwisay 20(2):I2s-)30, 19.1. Odsr aiww.f: Nalioa.l Irwitules of Health. From IAe tJep.rtntea of Biocbemi.lry, University of Qeor=i., Atheru. INiERACTION OF HUMAN a; PROTEINASE INHIBLTOR WITH NEUT1tOPHIL MYELOPEROXIDASE 1n earlies atudies. it has beea sbowe du1 a-l-praleituae iahibiar (a-1-PI) can be i.activMod br myeloperoaidase iw drc prese.ce of hydrogefoperoside and chSoride iow. Si.oe atyeloperosidne may be readily «kned from rre.uophils, its oaidative iw.cti- •tlio.ota-1-PI may indirectly resrv isatAnreemeM of proreolyuc destryction of Iwt 6sstrc, even in individuals with genetics2ly normal kvels of idtibitiorr. Tbe aim of tbia i.vestiptioa, dreafore, was to fwther iavestipte dre p.rsrnetcrs of the mye/oFer- oaide-modiated osidative inactivation of a-1-PI. These attsdies, as reported heR, dernorusste that dnxe is a direct dependence on the eoare.tratiat of a-1-P1 which coaeewtratio. of sbow 1.S µM. Z1rcte is also. depe.dence oa bocomes sa.radngrn dro concentration of H,0, to 93 µM, afler which increasing concentralions become iaicieaain6ly inAibitory. Chloride ion is requited for myeloperoaidase oaidative actioa, but dte carions Na' , NH,', or K' have littk effecl. There iss very sh.rp pH optimum at pH 6.2. under physiological NaC1 cance.tration. and approsimately half the r.re of inac/ivnion of a-I-PI occurs at p11 5.9 ar 6.5. Sodium dodecyl sulfate Sel ekc- aop/wnesis indicates that a-1-PI oaidized by either myeh+peroaidae or Nchlawuc- einindde is not different in sizt from native a- I-PI. When oxidized a-1-P1 is iwctrbrod with porciac elasuse. the a-1-PI is converled to a modified form of lower molocwlar weight. Amino acid sequence wlysis confirms these results in tballhe sequence is difkrem from dta of native a-1-PI. Overall, fsom this dau and other observariows reported here. it is apparent that oaidMive processes directly affect the inhibitory activity of a-1-PI modification of rhe n:active-site rneUtiowine of lhis protei.. 76 77

i 1 Matheson. N. R. Wong P S. Schuyler. M., and Travis. l. Aiew Annnery 2/1(2) 771-736. 1911. OtAAer rary..r: National Instnutes of Ikahh. Fnwn the fkpartmenl of Biochemistry. University of Georgia. Atheas. INACTIVATKN! OF IIUMAN PLASMA a,-PRO'iE1NASE INIIIBIT(Nt BY A MI--1'AIl.()P1tOTE1NASE FROM SFJtRAT/A MARCESCENS This sludy was undertaken b Mvestigaae die irNeraction of hutnan plasma a,- pateinase inMbitor with a meullopvseinaae iaolMed frorn Serrata iwarccrcens. Re- suhs presented here show that the insenctioa of a,-proteinue inhtbitor with the Srrroria proteinase caused a rapid deexnse in inbibitory activity lowards trypsia which was both concemrstion- and time-deperdeM. To determine whether the inactivation had occurred by IimiNed proteotysis. proteinase-a,-ppoleinase inhibitor incubation mtstures were denatured and analyzed by SDS-Polyactylanide tel ekcltuplMxesis. These results show tMt the prvteiwaae had npidly converted a; ppotexuse inAibitot into an inactive foret of lower mnbcular weighl (IA,000 for the modified form vs. 52.000 for the native inMMta). Amino termind sequence analysis indicated that the interac,we of the inhibitur and en:yme wu at the reactive sile of the inhibilor. with pcpude•bund cleavage resulting in the inactivation. This process may be important in, nocrdic prvicesses occurring during b.clerial infiltration of host tissues. Virca, (] D., I.yerly, Q.. Ktrget, A, and Trovu, l. aiarAiinica t•t BiapAysiro Acta 704 267-271, 1912. Other sryaf: Nriond Inuuutes of HeaNM. Fom the [7ep.rtment of Biochemistry. University ofGax6ia. Athens. and the Ikpart- nrat of Microbiology ard Immunology. Bowman Gray School of Meriicine of Wake Forest University. Winston-Salem. NC. . VI. Inrnfunoloa,p and Adaptive Mcchanicros CHEWyTACT1C ACTIVITY GENERATED FROM THE FIhTH CCtMPONENT OF COMPLEMENT BY PLASMA KAWKREIN OF THE RABBIT Kalliluein puriRed from rabbit plastun has been shown in the study prescnted here so generate cAenwtactic activity for rabbit neutroQhils Gom the nbbit's 6fth compo- nent of complement (CS). The effect on CS appeared to be due to kallikrein itself and nw to a c~u+tamonatin6 enryme, because i1 could be inhibiwl by anti-kallikrein 1~ or by >,vyl.can t.yp.m mhoAdfN to the ume eatenl that kinin 6encratwts by the same 78 I kalliktein preparatinn was inhibited by these agents. The chemutaclic response was cunsislent with Ihc generation of a CSa-like peptide from CS, because the effect could be prtatly nhtbeed by carfioaypcpWase N and was retated to the generation of a small (-11,000 mwd wt) fragment uf CS. In conlrast, no chanotactic activity could be demunstrated when the :ymo6en prekallikrein was tested with CS wtder identical c.nditwns. t:'hemdactie activity was also generated when rabbit CS was incubated with the 110.000-M, forns of activated Hageman facttr,lrypsia or EAC42). No chemo- lactic activity was pmduced when CS was absent from the incrbalion taiautes or wken intact CS alune was assayed. In stnn, ahese resrlts suggest dte esislence of a novel interaction between the Hageman factor and eompkme.l systems whicA may have biak><ical relevance. Wiggins. R. C., Giclas, P. C. and Ilenson, P. M. (CocMowr, C. G.) lournal of E.periwwntar Mesicinr 1J):1)91-1101.19e1. OrA.i nqprt: Nrionnl lnstitutes of Ileallh and the O(Ace of Naval Research. From the Department of ImnnntuPalho{MRest:areh Institute of Scripps Clinic. La lolla. CA.; and the fkpartmea of Pbdiauia. National lewistt Hospital. and Oeprt- meats of Medicine and P.tlto7oSy. Univasilr atrolaado Medical School. Denver. GUINEA PIG HAGEMAN FACTOR AS A VASCULAR PERMEABILITY ENHANCEMENT FACTOR In this anong attempt b ascertain the biological role of the contact (Hagemnn factor) system. HaAetoan fator was purified imtw Btrinea pig Vlasms by successive cohwr chromatoAraphy, and an active Haeenw (aclor. J3-HFn, was prepared for study. The Rttinea pig HaRemae factur appeared Aoewllenows as a sinAk<hain pro- tein on tsolyacrylanwde gels in the presence of sodiwn dodecyl sulfate and fl-mercaP- tneth.nol. Amino acid coatpasilion of the Rttinea pig Hqenta" factor was simil.r so ttnt rcported tor hnm.n, bvvine. a.d rabbit HaReans factors. The pwiAed Ruinea pig HaAennn faclo., as weN as Sttiaea pig Pla.mn, showed strong clotting t carroctiowane activity in Hasemr-faclor-debeint Iwteean plasma. The activity could be blocked by the IRCi ftaction ot swliaewrns against guinea pig H.Ranan factor raised in rabbits a a Ron. The concentration of HaReatr factor in guinea pig Phnraa was dettxmined to be 120 µAlnJ by quantitative radial iatnwnodifftaioa assay. When S-IIFn, the 2t,0DU- ddton active farm of HaZemna facbr, was prepared fiom Rniaea pig HaRaer faclor by ueatment with plasma kallikrein, 0-HFa catued an increase in vascular penaabi/- ity when injected into guinea pig skin at concenuations as low as ) a 10• M. 'iltia increased petnrcabilNy was shott-IaslinR. and nhe permu+bility enhancement activity of B-HFa was bthibitod by ppetreatment of P-11Fa witA diisoprophylAuaophotpAne. According b the authon, it nuy be conchdcd, dtereforc, that active Hasanan factaar in tAe interstitial space of guinea pigs acts as a vascular permeability tactor of for Sreater putency than bradykinin. . Y.marnoto. T. and CocArant, C. G. Aa.rrkan lorrnal oJParAoloay 105(2):164-17), 1981. ot6er say/orf: Nationd Inuilwes of Ikahh.and 1he Office of Naval Research. From the Ikpnrttnent of Invnumvpatlrtluily, Research Instilute of Scripps Clinic, La blla, CA. 79

M(N)UI.ATION OF POKEWEED-MITOCEN-1NDUCED IMMUNOGLUAULIN SI:CRETION BY HIIMAN BRONCIIOALVEOLAR CELIS The effecu of pulmonary alveolar mrrophates (PAM) on immunu6Mbulin (16) secRlNNI were inves/i6Ned, using aulololous peripheral bhwd lymplwscytes as the indicator p.K+ulatiun and pukewced milo`e. (PWM) as a rasnucyte-depenJeM puly- c1oMSal B cell aclival.u Bronchualveolar celis (BAC) frorn seven nonsmukm(I subjects suplxessed the resp.wsse to PWM by u.lnctionaled autolotaws peripheral blood nsonueuckar cclis (MNL). whercas lew eoareatrations of BAC partially teconstituted the response of monocyte-depktod MNL so PWM. Thus, it could be seen thm BAC could mo.Wlate PWM induced Ig sectetioa in different ways, depending on the prcs- ence or absence of monocytes in the mononrckar cell population. The suppressor activities of BAC were siol a"MOd by prior irradiauon and were only partially rcvenod by the addition of i.donseth.cM b tJm cukures. IWwever, prior disruppinn of BAC compktely abolished tlseir suppresslve h.ctions. Suppression of PWM induced IR secretion is probably mediated by i.t.ct, r.dio.esirrt PAM. [awrrncr. E. C.. Tlseodore. B. 1. and Martin, R. R. Anrrrican Rrvww oJ Rr*borary Dlsrasr 126:213-252, 1982. OOther s.A.rf: Amrnca.l.unR Association and the National Institules of Health. From the Rockwell Keough Pulmorsary Ielwwnology Laboratory aud the General Clinical Research Center of the Methodist Hospital, and the DepsrtmeM of Medicine, Baybr College of Mcdaine, Ilousbn. DEFF,CTIVE IMM(INOGLOBIILIN SECRETION IN RESPONSE TO POKEWEED MITOGEN IN SARCOIDOSIS Several previan studies of sarcoidosis h.ve indicated a dichtomy bnween en- hanced humoral immsme functions clinically and defective in vitro B cell responsive- .ess, which suggested some disorder of iertwnoretulMion. ln the study presented here, it was found that polyclonal irmNteoRlohulin (IR) secrclory tesponse to poke- weed miloge. (PWM) - a plant lectio which teqrites Ue presence of both marocyles and T cells in order to trigger B cells- wss defective as well. Specifically, in vitro inrnunoregul.linn of 1t secretion was audied in 21 patients with sarcoNkssis. While peripheral blood monomrckar cells fmm.ore+al individuals respoaded to PW M with a 14fold a treaser increnrat in IR-socrHi.g cells, cells from ssrcoid patients failed to respond b PWM N sny concentration employed. Morc rnonocyles were found in saceoid monanuckr eell prcparaions (N. t S 2.0% vs 30.1 s 1.1% in normal donors). bu1 removal of snasacytes improved the response 1o PWM in only four patients. Monrnwckar ceRs from sevew of 19 patienu supprcssod IR sectetsnn ia cots.ltures with normal donor cells. Patients eshibiting escessive suppressor cell function were ohkr, with longer standing and less chnicslly sctive disease than non-supprcssir `patients. Monocyte removal reversed the suppression iy only four of the suppressor patients. but escessive suppress,w rrrsmryte function was IMn demonstraled in two sarc.sid patients wMne crlls .nitu.lly dw1 rwM suMwrst Ig secn:bon when cultured with normal cells. Whik Uir ..w..uir.l..Rr.1 .klr....n sanoalnu may Ar c.NSy.k.e l.rlcr..l(enows and dynamic, these data suggest that suppressor rnorsocytes, when prcsen: in sarcoidosis. may Aave devckoped secondarily. Lawrrnrr, E. C. rt at. C'finiraf onJEsprrirnrnrallesmrnologv 19:96-101, 1952. Other s.ylert: American I.un6 Association and the National Institutes of Ilealth. From the Rnckwell-Keou6h Pulmonary Immunology Laboratory and the General Clinical Research Center of The Methodist Ilospilal, and the Department of Medicine, Baylor Colk`e of Medicine, Ilouston. NEUTRAt. GLYCOSPIIINGOUPIDS OF HUMAN ACUTE LEUKEMIAS A anethoduksgy was introduced i. this study which combNles the sensitivity of high performance liquid cirurnMo6rrphy with the speci0cily of eso- and endn6lycosl- dases to study the neutral ilycosphinpsllpids present in the malignant cells of 10 pnieass with acute leukemia. Results showed that acute kukenua cells contain very link or none of the more corrmpka neutral 6lycosplwnRolipids dw ne found in normal kukocyles or chronlc kukearoa cells. Lymphoblasts, in prticwlr, a+e rich in neutral tlycotalthin6olipids with only one or two csrbahydrste unils. 'IUe most sisnificsr i.dine of this study was Ihat, in contrast to.ormsl krkocytes aed chronic krkemia cells wlwch have a single predominrN tetraosykeramide species, acuae kukernia cells (9 out of 10 patients analyted) were fowd to have sig.i/icasl amounts of both gbbo (GsINAcol Y 7Gala1 Y IGalol Y IGkoI Y kentnide) and neolsclotetrsosy- kerarnide (Gal(il Y IGkNAcj5l Y Xh1JM Y IGkol Y keranide). These results Mdicate that the canposilion of aetMrd RlycosphislRolipids IM scute kukemis cells difkn significantly fmm that found MM normal ar clrowic kukesnis cells. Iee, W. M. F., Weatrick, M. A. and ifarArr. e. A. The Journal aJelolotkef CAnnistry 2S7(17):10090-10p9S, 19/2. OrAer sap)rort: National latilules of tkaldt, National Csncer Institute, Lntketuia Research Fou.datio., and Cawer Research Funds of drc Univetsity of California. From the Cancer Research (saitWe, University of C.lifonds. San Francisco. t GLYCOSPf11NGOLIPIDS OF NORMAL AND LEUKEMIC NUMAN LEUKOCYTES Studies on neutral Rlycosphin`olipids and pngliosidcs of aormd aed kukemic human kukocytes were reviewed for this prcsenlation. It can be seen here thr two methodological appnsaches have been used w determine the strtrcture and distribution of 6lyarsphin6cslipwls amonR human kukocytes: (1) Mose thn utilize chemical and enzymatic tuols w delemtine the complete strrcture, and ( 2/ those thn sdy an indirect assays of detection The former methods allow one to assign a unique structure to each corepound, but they have twodisadvanta6es: (Il they teyuire rdtlively IrRe quantities of materials and (2) minor components may be lost during the process of prepring hoer.henuus ccNSSpoumis for analysis. Indirect methods which have been used are sensitive sad rapod, and allow (w.c NscornPare easily several saepks, but dhey have die wl , KI

I disadvantages of being indirect, with stnrctun:s assigned on the basis ni cornparison with stand,rds. Esammed f4w this review were: la) the 6lycosphinSotopNf conyrnititm of varwws kukocyte pqwlatMins, (b) the differences in 6lycosphin6i hpnls found among subsets of these «lls. Ic) the possible use of these compwnds as markers of differentiat..n. and (d) the changes is glycosphingolipid composition lhiq occur with kukemogenesis. MaArr. B A.. I.ee, W. M. F. sd Weslrick, M. A. Molrcular ard Cellular BiorArnslslry 17:11-95, 19t12. Other rayp..t: National Institutes of HeahY, National Cancer Institute. lrukemia Reserch Foundation, and Cancw Researd# Fwds of the Universily of California. Frum the Cancer Research Institute and Dep.rueent of Pharmaceutical Clamisrry, UniversMy of Cdifornia, San Frnneisco. NEURON SPECIFIC EN(1l.ASE AS AN IMMUNOCYTOCHEMICAL MARKER FOR THE DIFFUSE NEUR()ENDOCRINE SYSTEM IN HUMAN FETAL LUNG Neurow-sQecifu enolase (NSE) is an isoentysne of the Slycolyuc eswyme enolase, which was originally considered lo be restricsed to netarons but has recently been show. b occur in some APUD cells 1s this p.per, the localization of NSE in the diffuse sxuraendocrine system of human feW lung is reported. Speciricdly. NSE-posilive u11s, singly or in 6roups, were demonstrated by anlisera raised to human or rat NSE. Imnwnostainod send sections indicated thM NSE-positive cells could also eontain bombesin and 1« SHT•tike imrnunoreactiviry. At least three different cell types were identified containins (1) NSE. S1fT, and bombesin, (2) NSE and SHf, and (3) NSE alone. Aftcr close consideratiexi of the material presented here, it appears that NSE is. •seful m.rker of the neuroendocrine system in the Iing as well as rn otAer tissues. Aho, the lack of alterrsative simple and teliabk techniques capable of identifying both cells and nerves means that the iermmocylochemicallocaGLtion of NSE is a valuable tod for dre study of developnrent, physiobgy, and palioloSy, of this system. Whartow, l.. rolaA. !. M.. Cole. 0. A., Marasjoa, P. 1.. and Pearse, A. G. E. The lournd af Nisraebtntistry a+d CyroeArndtrry 29(12):1359-1364, 19i1. From the Department of Histochenrstry, Royal Postgraduate Medical School. Lon- don. England. and the Clinical Psychobiology Branch, National lnstautes of Health. Bethesda. MD. IeE-DF.PENDENT RELEASE OF LEUKOTRIENE C, FROM ALVEOLAR MACROPHAGFS Slow teanin` substances (SRS) have been shown recently to bc a family of peptidnfipds called kukouienes (LTC.. LTD, sed LTE,) shu we derived from arachi- donic acid and are potent bronchoconstricton /n vivo aad in vitro. In the study pe- sensod here, rat slveolar macruphases we shown 1o be the cells rtspnmibk for releasing thc SRS The ability of these mrcrophases to release SRS was tested initially by incubating cell suspcnssouu far 20 min with I µmol o(cakium wwwK twne A231t17, 92 I in the presence of S a K) 'M L-cysteine. la subsequent esperiments, cell suspensions w4k.re stirwulated with purified mouse oruwclunal uyi-DNP /dmilrophenyl) ISE anti- bcdy and INdP-human ><erum albumin. ResuNs of these esperimenls show Mn (1) rtl alveolar macropha6es release SRS when stimulated narspecifically by the cakium ionophore A231N7 in die presence of L-cysteine, and (2) I`E atwrbody and approQrire antigen cause alveolar manophases to release SRS kukotriene, LTC.. This demo.- atratios 1hN rat alveolar macrophqes release SRS by an IIIE-dependeM mechanisss raises the possibility that ISE-depadent rekase of inedislors by dvwlat ~nacsoQAa~cs may have a role in asthma or ather immunologically eredialed Iu.B diseases, R.nki., l. A. n ol. (Raynolds, H. Y.) Narwt 297:329-331. 1982. ~ Ur4r sayr.s: National lnstituks of Health and Ho(fman La-Roche loc. Fsan the Dep.rtmewt of Medicine and PAarse.eology, Yak University School of Medicine, New Havcn, and the )oh. 8. Piaoe Fotawdatiow, New Ilave., CT. MONOCIANAL ANTIBODY ANALYSIS OF COMPLEX BIOLOGICAL SYSTEMS: COMBINATION OF CELL HYBRIDIZATION AND IMMUNOADSORBENTS IN A NOVEL CASCADE PROCEDURE AND ITS APPLICATIAN TO THE MACROPHAGE CELL SURFACE In this sophisticated saediodolokical pper. a procedire is describeA whicb peatty simplifies the edlection of aowoclo.d antibody (MAb) libraries d'uested townd i.dividwl compoee.ts of canple: biological systems. Foa the study repoAed Aae, tesnovd of previously tecopimd aatigews widt itnnw.u.dsotbem coMwsm wr oa.- bi.ed witb cell hybridization in a cascade which restric/s the ieuwei:inR stinwlrs to p.eviorsly wnecoj.ir,ed antiBesu. Specitkdly, i. dds report a casc.de procedire wr e.plosed is co..edan with dte identification of (wdKr m.no4A.`e-specific wille.s. Pr•sitooeal esttd.te cell membra.es were dckqeM 1olrbiRrsd, and the previously identified coawnow kukocyte ssMiRest assd beal-slatble oafiBeo which we ahared witb peritoneal esudale celb and lymplwcyles wesm sawoved with MAb' immMO.dsaa bents before imm.niLtion for the hybridi:alion esperimettl. Removal of tfte antigens was confirmcd by r.dioirmrwomay and by the sesological response to immtriritan. Serum antibodiea b specific aMigens were also measured to compare die efficacy of dris procrodure to inrartizatiow with eitAer wlole cells or MAb-cosud cells. Two previous/y arknown macsvph.ge-sqecific antigens of 32.000 and 110.000 lµ were identified here. According 1o dre autlwn, the pracedure can be eaended by arranging hrlher immuno.dsorbenl depletions and cell fusions in a cascade series and is roadily applicaPk to the manockseal antibody ar,dyses of many other mulucomptment biolog- ical compkses. Springer. T. A. T16e Jownd oJBioloticol Chemistry 216(s):3R33•3i39, 19111. OrAtr r.yr..it: U. S. Public Health Service. From the tkp.rUeent of Pathology. Harvard Mcdtcal School. Boston. 91

MAC 1, 2. 3 AND 4 Ml1RINE MACROPHAGE DIFFI:RENTIATION ANTI(:ENS IDENTIFIED BY MONOCLONAL ANTIBODIES The Kwhkr-Mslsiein myeloma hybrid lechnique, which can enable the iscdation of a monocwwul nwthndy recognizing a sin=k arMigenic deletminant from an initial highly conpka anli6en, has given great impelus to the analysis of cell surface com- Ple.ily. In the paper presented hert. wort done in the wlhor's laboratory w.ins this technique Wr the idemtficalinn and study of nrcrophage antigens is reviewed. Fhwr antiRens -- Mac-1-2. ) and 1- have been idenlibed by the correspondins moa wsckmal rwibodies, M1/70. Ml/)1, Ml/)/, Ml/iA and M3137. These antigens all appear to be on the macraQhate cell surface on dre basis of Mrorescent and "M-Lheling. "S- mclhiwmrne incorporation into the polypeplides by the dterent fraclion of thio6/ycol- lale-irwluced perNorseal esudale cells also suggests these antigens re synthesi:ed by macropha6es. The fout different anullau defined in these studies are pa:senl nn rnacrrxrlsa6es, bu1 not lynsphocyles, demonslrssag dse distinctiveness of mM rophaae cell surface archiNeclure. Cwren11y,1Ae espressiow of these antigens on mact.+Phases induced by other means aed in different ruwornkal locations is being investistced. The munoclwul antibtdees re also betng used as probes to inhibit a panel of marrophase functwms In this way, it should be possible so link the molecular struchrres csescnhed here with specific macropha`e cell surface activities. Springrr, T. A. In: F<hsler. O led / Ilrrrrotrnriry oJnrononrr/ror pAotoryart. New Yort: Academic Pnu, 1980. pp. 17-I6 Other srp/etf: U S Public Ikalth Service From the Dep.rtmeM of Pathology. Harvard Medical School. Boston. A SHARED AU.OANTIGENIC DETERMINANT ON Is ANTIGENS ENCODED BY TIIE I-A AND I E SUBREGIONS: EVIDENCE FOR I REGION GENE DUPIJCATION 11 has boen known for a while that die I relion of the major halocompatibility ooml+ka conlains genes that conavl immune response and imnwne suppression to oeAain antigens and dif fenenl delerntinaau. and studies on these genes have led to the de6nition of a nuesber of I subregions. In the study presented here. two rat nonoclonal antibodies (MAb). M51114 and M7/111, which have a very unusual type of crossreac- live specificity for mtrine I region prodocu. re chrscierined. These MAb detect pnlymorplsic determinants present on 8 cells and activated T lymplsocytes from mice crryatg the H-2•.11-2'. H-2'. H-2', and ti-2•h.plolypes but uo1 from mice carrying the H-2' or H-2' Aaplotypes. AMigenic sMe tMUnber delerminMions showed that the /wsitive haplotypca can be divided into two groups. Mice bearing the H-2•, H-2•. and 11-21 limplwyaa espress a high number (40.000 to 110,000) of antigenic sius pcr B lymplwr cyte, and MAb plus cornqlement can lyse 8 cells from these mice. In conaast, mice bearing the H-2• rd H-2 hapkMypes eapess a bw number of antitenic sites. Spleen cells (mm mice carrying the latter haplotypes re nollysed with MAb and compkment. Genetic mapping demonstrated tluw high and low espreuion map to the I-A and 1-E su"ions, respectively The MAb detect an Ia speeificily on 1-A•,1-M,1-E'. and 1 P nroktuks 11Kse /dNtr.MNom were confirmcd using trveral dillerenl eaperimental .p/wu.che.. i e, cyuw4s.wNy, /lwncscen/ sutning, competitive inhibition e>t MAb bindins, and 2-dimcnsNmal gel ekctrcqiraesis of immumMxecipitate.. Resulls of tAis study provide immunologic evidence (or humolosy between I-A and I-E antigens, and hence kw senc duplication within the I region. Bhaltacharya, A., INwf, M. E. and Sprinarr, T. A. TArlorrnul oJlwmsunolosy 127(6):21/1t1-2195, 1981. OrAer supperr: U. S. Public Health Service. From the IAborawy of Membrane Immunochemistry, Sidney Farber Cancer Institute, and tlse Ikpanment of Pathology. Harvard Medical School. Boston. NATURAI. KILLER ACTIVITY IN THE PERITONEAL EXUDATES OF MICE INFE(TED WITH [.IS"."RIA M(M/OC1TlX:F.NES: CHARACTERIZATION OF TIIE NATl1RAt. KILLER CELLS BY USING A MONOCLONAL RAT ANT1-MURINE MACROPHAGE ANTIBOOY (M1/70/ Natural killer ( NK) cells we monotMrckr cells of disqt.ted lineage dsr kill certain destructive cells in the body. In t:ae paper presented here it can be seen dul infection with Usuria wwsocyrosrnrs Q..hCI kads to the 6encration of NK activity in periloncd esudates. Specifically, m.xinwm eRpresiioe of NK activity Rn1 occursed on day two and remained high until day sia after initial eaposune to LM. When nylon wool nonadherenl peritoneal esud.te cells were esamined by a single-cdI cybtosicNy aa- say, the wmber of cells binding to YAC-1 target cells increased after infection n did their ndlvidtul lytic capacity. A nsonockfla.l nl awti-mtusne toacmph.te arrbody, (MI/70), previously shown 1o rocoproe human NK eeBs, can be used also as a wmker for wurine NK cells. Utilizing MU70 and the Buorescence-activased oell soner, seke- tion of MI170-labekd mononuclear cells led to the esricAnrc.1 of both NK and anti- body-dependent cellular cylolosicity. These MI/74Positive celh hd a distinctive morphology and conuined parwks on wright-Giawsa staining. They were not Phago- cytie, did not contain nonspecific esterase, and lacked surface 1-M, /SM delemsisaxMa, complement roceplon. and high levels of Thy 1.2. Holmberg. L. A.. Springer, T. A. and AtJt, K. A. TAr Journal oJlwrwrnolop 127(S):1792-1799,19d1. OrAar aayrar*: Naliond InatNules of Health. From the Department of Prhokogy. Harvard Medical School, Boston. , MACR(W/IAGE DIF7FRENTIATION ANTIGENS: MARKERS (lF MACROPIIAGE S(1BP(WULATK)NS XND TISSUE l.(1CALILATION The characteristics of four distinct antigens which re presem on macroph.ges, but nollymPhucytest we reviewed in this Paper. In addition,lwro applicrions of nwi- macrophaSe rsrwsock>rsal rea6ents-their use in pherwtyping snrrophade subpcqu- Ialions and in idenuficnans of macroPls.6es in tissue sectioa-aro described here. The wsajcx soctions of this paper are devoted to MAC-1 ANTIGEN, IMMUNO- Af)SO1tBIiNT-CELI. IIYBRIDIIJITI(Nd CASCAt>E?S, MAC-2, 3 AND 4 ANTf- 94 • 95

GI:NS. anJ IDfiNTIF1CATION OF MACROPFIAGES IN TISSIIE SECTIUNS BY IN/)IRECT IMMl/N011.UOR13CEN(-F. Overall. four macmphase anugens with distinct Mr and tissue dwnbulMm are wkntified in this wnrt. Two uf lhese. Mac-1 and Mac l, are synmhesircd by all macrophate subpopulatwsns examined thus far Iluw- cver. Mac 2 seems w be prefereroiafly associated with 1hw1glycollate-elicited penlo- ncal macny+haRes la anti6rns show a different Pattern of expression Therefure, macnK+haRes can be defined into subsets with distinct aMi6enic phenodypes, as is the casc for lymphocytes. Springer. T. A and Ilo, M- K. In Mitchell, M S and (kil6en, H. F. (eds-): Hybridorwai in Cancer [)ia6rw.sis and T.ranwrrr. New Yort. Raven Prcu, 1981. ppp. 35-46. (kArr s.yp..t: ll S Public Ika1th Service. Fro.n the 1)eprtnrnr of Padsuk.gy, Narvard Medical School, Bosto. RAT ANTI MOUSE MACR(W4IAGE MONOCLONAL ANTIBODIES AND T11EIR USE IN IMMUNOFtU(MESCENT STUDIES OF MACROPHAGES IN TISSUE SI:CTIONS The characterisuics of five mnnoclo.al antibodies (MAb)1o macropha`e antigens are summarized in this paQer, which also contains a deseriptiow of the use of one of these M Ab for the localization of mano~s in frozen spkew sectiasu. OI the five rsl ero.ackitsd anttbodhes to modse macroQM{e sssifaee antigens that were dr.vebped is the wtMars' labrxsaorp by the mycbma-sploets cell Aybrid technique of Kohkr and Milstein. MI/70, whech recognizes a phftocyle-specific anlijen. Mac-1, is the most extensively studied Two other rwebodies. M3/31 and M3/31. pecipitasc a polyPeP- lide ternsod Mac-2. which is also characterized kse. In Ihe related stwfy, an1i-Mac-1 was used to stain rnacropha6es in spkes sections because Mae-1 seems lo be e.pressed on macropluges irrespective of Mrir uale of differentiation and aclivation To allow alignment of areas with Mac-I' cells with Tdependenl n=n of the sploen, adjacent sectiom were stained with M3J49, a. ami-Thy-1 MAb. Resuks showed Iha/ T lympho- cytes in the periarte riular l ymph.tic sheath are intensely stained by wi-Thy -1 M Ab . In corwrw, few, i(.uy. M.c-1' cells can be seem in these T-dependerM areas. Tlhe simple method for the bcaiization of macrophages described in this pubbcatias can be easily extended to oUrer swdies. especially with a.wi-Msc 1, which stains all macropMte wbfsopulaiiotn examined so far. 1n view of the vast body of information that can be gained fr+ae anatomical localization of lymphocyte strbpopulations. these studies dwuld prvvide much insi6hl into dte function, differentiation, and ontogeny of nucropha6es. Ilo. M•K. rd Springer. T. A. In: Nanrnerling, (I , Hammerlins, G`and Kearney, l. (eds. ): Monocloaal anribodiri and T rrll Aybndonwt. New York: Elsevier/North Ilolland Biomedical Press. 1981. ppp 3361. OrArr /.yPwrt: U S Public Heahh ServKe. 1-nvn the I.Ir+.»nry 4d MrmM.ne Imswmschemistry, Sidney Farber (-.ncer Institule. / Iar.arJ MrdK .! ti. A..el. 1t.sion MAC-2, A NOVEL 32.000 M, MOUSE MACROPNAGE SUHP(WI/I.ATI(NN-SPECIFK: ANTK;EN DEFINED BY MONOCLONAL ANTIB())IF:S The billchemical cfuractcrizalicm and «ll distributinn of a 32,t100 M, anlige., lenned Mac-2, are presented in this paper. Mac-2 is synthesized by and expressed an drc surface of thinslycollate-clicited macrophages as shown by I"SI-rnelhionine aau '"I labeling. Unclicited pcriloneal m.crophases and macropha6es elicited by prasease peptone. Con A, LPS. and Liurria n.awxyrosrwrt ne either only weakly positive or negative. Thereforc. Mac-2 expression is induced only by strong i./1ammNory stinwli and appears specific for momonuckr ph.6ocyle subpopulatiows in a distinct stage of ddkrenlialion. ResuNs of saturation binding eaperiments show that Ihioglycollne- elicite+l macrophages espess 1.7 s/0' Mac-2 sileslcell. Thio=lycollale-elicited macro pha6es are slmngly absorylive for "'1-labded M313t MAb. Cell suspensions frorn sPleen, boee m.rrow,lhymus, and peripheral lyniph.ode dft > 991L Mac-2 negative by wnnwntsfluaescent /low cytomelry. In contrw, 1hioillyeollate-elicited macro- phages we > 96X strongly pusitive for Mac-2. SDS-PAGE of 1"SI-methionime- ktbekd Mac-2 shows that thioillycdlresliciled <r.nopbaps synthesize 14 to 30-fold more Mac-2 than other peri/o.ea1 macrophage srbpopulaioas, whereas all hPea of Perilorxd manophages sy.llrcsize sad express on their surtaces similr srnowtts of the Mac-1 anlip.. Mac-2 antigen is, tAenefore, induced in macrophages only in n:spowse 40 specific difkrewtirive signals. Ho. M-K. ssd Springer. T. A. The latrnal aJlmiwrMoloRy 1211(3):1221-122t, 1912. 0&+r s.r?ws: U. S. Prblic IIeaMA Service. From the lsborrory of Membra.e Innrrochemistsy, Sideey Fuber Cancer Lstilwe, Bo.low. OPfTOGENY OF MURINE MACROPHAGES: FUNCTIONS RELATED TO ANTIGE'ld PRESENTATION Immadwe macrvph.Re Gt.c/iow contributes 1o the i.aessed susceptibility of sets- *Mes 1o infection. /.1his Papet, Me imtuls+rily of .eorulal reacraphage fwrction is dissected into antigen pesesNtlioo and tAree different ef fecwr components: cylolosie- i1y. antiken uptake sssd c.t.bolism, a.d (he prodretioo of the lymphoaimul.rary enloce'k waerkrti.-1 (also called dtymoc)ne mikKesic protein or lymphocyk-a- livasis+6 factor). The uptake aad calabolism of °M-labeled Linrria wrrnoc"rwrs were oqrivakM in aucrophases from aduA and neonatal ruice. Ikswever, awer.ctiosy be- Iwees macnspha6es fmm neoeales, heM-kilkd Lisarta orgaeisms, and immune T lymphocytes were impaired. and no cyacidal macrophages capable of killing tumor cells were Senerated. Previous studies with cells from rduk mice had established that the development of cytocida) macnsphages rrqvired la-bearing. aiaiRen-preseming macrophases and his/ocomp.tibiliwy at I-A between auavphnes and T cells. To circwmvcnt this requiremenl for anti`ea-presentin` macrophqes. an assay was usod in which IymplMikine was added directly to the m.croph.ges from neonates. Strong eylocidd activity resulted TMn, these studies showed thr aunophages fio.w wo- wes present antigen pMKly but can acquire cylocidal frnctioat provided dW the mood 86 87

II fur amiKcn presenting funNion is bypassed. Similar conclusitxn were rcarhed Gor the fetRYNM of itMctkukm-I In essence, all the dals presented here IndK'ale that the iny+airment of s number o( nucrophs8e functions in the nnmates is due to a reduced number uf Isrpoawve marnopha8es. Lu, C. Y arnd Unanut. E. R. In/irtiwt and linmunity 36(1):169-173, 1912. Other sryp.rt: National Institutes of HeaNA rd the March of Dimes. From the faeprtment of Pathology. Harvard Medical School. Boston. C()NTR(>l. OF MACROPIIAGE Ia EXPRESSION IN NEONATAI. MICE- R/KF OF A SPI.ENIC SUPPRESSOR CELL As reporsed in tbis pspa, the cowtrol of tw.craphs8e e.preuion of I re8ion- associatcd anu8ens (Is) in neonMal wtice was sAdied by compsria8 responses of neonatal yd adult mice to irrr+wrne vs. .owinratwe stimuli. AdrtNs generated perito- ned e.utLses rich in (s-hesritg m.csopAsges in response 1o i.p. injection of live Lirreaa mr.ocyrosrw.r. Lrreria-immune T eells, srd heM-kilked Listrrio, or a solubk me.hator termed macrophs8e 1s-reenNinB factor (MIRF). Neonates failed to respond ttt these stsmuli In coMra>t. both neonates and sduhs generated la negative perMOneal esr.dstes when stimulated with thio8lycolIale. There sre three major new points thal csme out of these stud.es:ll) neonatal mice na only have a defect in their basal nwnbet of Ia positive phsgocyles but also fYl to respond to the immune stimuG Uw generate esudates ennched foe these cells; (2) there is a suppressor system operal- in8 in the neonste capabk of significantly dsmpcnin8 the recruitment of Is-prnitive mscrophs8es-thn suppressor system b also operant in sane sduh tissues such as boae murow and the penauiesl csvity, and ()1 the suppressor mechnusm tewtlves, at kasl, the phsgocyte system by way of an mdonsethacm- and asptnn•sensMtve step. pvenll. N•ppears that this phago[ytK hne antaRgulMes /ts surface esprelsNM1 of Ia it1 bolA neonatal atwl adult mice Since this mechrnism becomes particularly puimed dwing early devek,pment, i1 could contnbtMe to the lack of imnrruty dunug onk><eny. Snyder. D. S.. Lu, C. Y. and Unenrr. E. R. Tltclotdnal aJIaowwoloRy 1211(3):11311-116S.1912. Oder seK..t: National tnstilutes of Heahh and the March of Dimes. From the Dep.nmen( of Pathology. Harvard Medical School. Bostos. SP(NIfANFAUS T CEL1. LYMPIIOKINE PROD(K'il(NJ ANI) f:NI/AN(*ED MACR()PIIAGE 1s EXPRESSION AND TUMORICIDAL AC1YVfIlf IN MRI: Ipr MICE Selected mscropha8e functions in MRL/Mp-Ipt/spr (MRL-Ipr) mice were evalu- red for thn repon. Specifically. tMoe tnscrvphs8e funetiuns were sludied in MRI: Ipr • mice with auloimmune IympMoprulifersuve disease: surface espression of I re8Kn- assrriaed Ils) antigens. tumnr cytotosicity, snd interkukin ((II. 1/ pn.Mrctiun. M/lI. Ipr InnY hM/ a s/6n/hc.nlly InCteased represen/a1MN1 of Is positive macaiphases m \ in the perikmed cavity, compared 1o n!1 normal straiws of mice. in order to atrdy the basis of tAis iwcrease, thymocytes or spknocytes fnnre MRL-Ipr mice were trar- pl.mcd atrap"ltonrafly inlo nonn.l nuoe. 7Bree days 1s1er the recipient mice lud peritoneal esudates rich in 1a-posNive m.croph.ges. The cells which induced dtw respo.se were T cells which elaborated a lymphokiae responsibk for the mcrriunent of la-positive eucrvpha`a. 1n previota studies fram this labarawry using mioe. lyt.phokita was soctelod only following the ieletac(iot, of inrruae T cells with anti- Rat. The resident m.croph.Res of MRL-Ipr mice were aclivned and killed (weot cells if triggered by aa interaction with baclerul products, eves, without 1he sddtttos of lymphokines. Socre(ion of IL-1 was normal. Results indicate dW (lre diseased MRL- lpr urce we ch.rscterinod by (i) activaed T cells t1t.( spontaneously seae(e m.cro- pha[e stitnulrory rnobcules, snd (ii) activated tnacroph.Res thr show both an increased espresswtn of Ia and lymphoki.e-isdt:pe.de.( triggering of luruoricidd activity. Lr, C. Y. and Unawt, E. R. Clinlca! la.wrnobry m.d ImnwwoparAdoty, 25:213-222, 1912. Or4r s.Wa.f: NationallWiuu(es of Health and the Mach of Dimn. Fiow dte Deprtarc.l of Pathology. Harvard Medical ScAool, Boston. YI t. Bpideneloto6y MORTALITY IN MIDDLE-AGED SMOKERS AND NONSMOKERS The stlatiatn of cipsene smoking /o ttstxulity was assessed in a.11-yest fdfow- ep -hdr of 4.004 mn nd wome., 33-Sd yeaa of aRe, who responded so tvRinB b have rtwNipAric heaNh checkups. Acc+owNieB for di mher eltarac(eristics. both indi- vidrally and in combi.rion. faikd /o elirninsle (he association of smoking with sixwlity fn+m all causes or with ma(dity from coronary hars disease. The smoker- to-nonsmoker mortality ra(ios. crude and adjusled respatively, were 2.6 and 2.1 far aB causes and 4 7 and 3.6 for coronary heart disease. 'Lbis analysis did wut support the cowMerhypothesis that the associaliono(ctgaretue smoking with morWity is secondary 1o some underlyine chv.cteris(ic. Frirdwnn. G. D.. Dales, L. G. and Ury. 11. K. TM New EntlandJorreal aJMeJicinr .)(10(5):213-217, 1979. Ofber sayp..f: Kaiser F.wndation Research Institute. From the Ikpartmrnt of Medicsl Methods Rexr.h, Ksiser-Permanentc Medicd Care plosrsm, Oakland. CA. 89

CHARACTERISTICS OF SMOKING-DISCORDANT MONOZY(X)TIC TWINS The Kaiser Pernunenk Twin Registry of Oakl.nd, CA, contains o.cr 8(100 pairs of Iwins who voiunteered thea Iwticipalion in a prv`ram of inedical nae srch on twins. In 1977 and 1978 a large health yuestiorrsaire was mailed to all twins aged 18 yean and over. AfKr the first nudrn8 in /une 1977, a reminder postcard was sent in (k lithcr 1977 w.orrespondrn8 samc-ses Iwins. In this questionnaire, each twin was asked about his or her own snrskin8 habts and those of his or her colwin. In the study presented here, (he snw+kmghabts u1 the J) cigarepe snwken in the smoking-discordanc monozygotic (M7.) femak pain wese compaed witA 1Aoae of the 205 other M7Z kmak cigarette snwiken Large rd sutistically sipi/kaM di/krences werc nnted in sorne measures of smukin8 rnlcssity. Ciprene snsoken wkro Ind an MZ nonsmokin8 colwin tended 1o stan smoking Incr and to smoke kwer cigarettes. Thn may esplain, in pan, the smalkr drf keence in CND occwrewee Aehraew smoken aa d nonsmoken within smok- in8-discordawl twins than belweew snaken and uowsmoken in Ihe general pnpulNion. Additional comparisons were also nnde t+ehvee~ /he discordant snwien and their nonsmoking cawrns. Results of these eompuisns show tIW, with regard to coronary heart disease (C ND) risk faclon, the diacordrM smoken wete leaner rd consumed mmue alcohol than their t.onsnsokinl cotwi.s. While these traits we associated with a lower risk of C1f1). snr+ken also lended b be less educated and reported less esercise and concern about physical fitness. eonsistent with higher risk. In conchnion, the data ow a hmited number of smnkin8discordant kmak MZ twins suggest tYat, even with genetic idc.trty, twins who drf ler in one characteristic may differ in other chracteris- tics rekvant to the outcome under eonsideration. Frirdnson, G. O rl at. In Gcdda, L, Parisi. P, and Nance, W. E. (eds. ): Twin RrrrmrA 3: £pidrminlo8icol andChnirof Strdirs, New York: Alan R. Liss, lne., 1981, pp. 17-22. OrA.r s.yp+rr: National lkan, Lung and Blood Institute. From the De9artmenl of Medical Meshods Research and Department of Medicine, Kaiser-Pennunente Medical Care Program. Oakland; University of California. Bcrke- ky; and the Department of Epidemiology and Intersutiorul Ikalth, Univenity of California. San Francisco. STRI/CTl1RAL ANALYSIS OF SMOKING, ALCOHOL USE. AND Pl:RSONALITY FACTORS IN MZ AND DZ TWIN PAIR RELAT11NdS111PS This study presents an eslension of the lestin8 of twin type compuabilily Io a muhivaria/e situation. UsinB data drawa ftom the Finnish Twin Co1Mat as the study pn/wlation, smoking clwacterislics, akohd use patterns, and personality factexs r>< tsrwrszyBrMic (M'l.) and dizy8rotic (D7.) twin pain concordant and dixurdanl for rn:- qucnt heavy akohd use were described. The effect of constraining awariance rela- twmships it individuals of either twin type was tested 1o estimate comlwahility of pauwise structures Als.r. the covariance relNionships in individual twins were corn- parcJ to th.xr od.inRlrasnf hor such analyces. the general rnndel for the estimation of hnru .nuk tur.l equ.u.-n hf nma.rmum hkcliMr.d melM.ds wa• aM+hcd hs Ihe ( uuush I+,n I .+..rs .r..t. .tua kr.uhs str..rJ that the dillcren.cs hctwccn hcavy- MI I anJ li8ht•drinkin6 individuals were nwssl ckar-cut in sinslctues They differed in the mean sawskin8 kvels, anJ the estravcnion and neuralicism scores. The same result was ohtaieed in 1)Z pairs discordant lur heavy alcohol use. In discordant MZ pain, Mrwever, thc smoking differences were observed but Me personality factor differences were seen in the neucdicisrn and life dissatisfaction sedes, suggesting that the basis of discordance in MZ twins may differ frorn that in DZ pairs arsd singletons. The propor- tion of discordant pairs was 291% i. MZ and )716 in DZ paas, less than the espected value. Mullivariate analysis of differences in MZ and DZ discordant pain confirmed 1he results from the univariate analyses, the relative si`ni/kance of neurvticism in MZ pairs beconsin8 wcaker and statistically nonsignificant. Smoking. newdicism, and life dissatisfaction, independent of genetic facton, seem to be indirawrs of pmcessec leading to heavy alcohol use. langinvaiwlo; N.. Kaprio, l., Koskeavuo. M., and Tarkkonen, l.. IRonrosulo. I 1 In: Gedda, (-. , Parisi, P. and Nance, W. E. (ah. ): Twin rrseorrA !: rpidewriololrcel adc/inicol srsdirs. New York: Alan R. Liss, lnc., 19M1, pp. 23-35. From the Department of Public NeaNb Science, Univeni~y of Iklsinki, Ilelsinki. CIGARETTE SMOKING, USE OF ALCQIIOL, AND LEISURE-TIME PIIYSK'AL ACTIVITY AMONG SAME-SEXEp ADULT MALE TWINS The selatiwmships of ciaasette smokin8, akahol use aal kisure-time physical activity among adult mde twin participants in a Fienish popvlatiun study we presented hert:. For this epw)emKdoBical study, queslionnaise responses fmm 1.537 nwnozySolic (MZ) and 1,507 dizy8otic (DZ) toak pain aged IN asd over weee analyzed in ternss of combinations of the three facbn and the relatiu.ship of these (acton to each otkrcr in rela/ian to the twin pair siluation. Todu this, thsee facwr analyses and an overall ckrster analysis wese carried out. Results of lhese tests showed that the physical activily, factor arcans wes~e almost corntartt with age. but Ihese was a decrease with age in alcohol cowswmp/ion; for Ihe srswrking facksr, there was a steady increase with age until 30-54 yean, after which a slight decrease occtsssed. TAe conelation coef/icieMs betwee.lhe fackrs in the whole series showed a high costelatiss between cigarette smoking and use of akolwrl, and small negative cnrrelatwsas for physical activity and ci8arenc smnkins and fix physical activity anJ use uf akohrsl. In ahe cluster analysis, eight cluslen weee found lo be stabk in Knrup-tesnwp situations with over 90% of nsemben remaining in the same cluster from oee analysis Io another. As to Iwinship, bunh MZ and ()7Z twin pair members were in Me ssme cluster setwh mae often than espected, hat the MZ•l)7e overall dd/erence was rcl,Nively small The hiEhest M'l.iDt ratins ol nhscrved w espn-ICd a9usterin8 rNes w.re in Iwrr clusten: A 1 cluslcr ms 7, which had Persnnc with a high nran deErce ul k iwrc-Iime physical aclivity; anJ 81 the very small chssler nn. 8, which had a very high snean sltrrlwd use. Kaprin, l., Kuskenvuu. M anJ Suna, S. IRonruaJa, 1.1 In: (adda. 1. , Parisi, P and Nanrc, W F't.Js I Tu-in rrsrtin h f- rpiJwni.doRirof w Jr'6ni, al srwGrs. New York Alm R 1.6%. Inc , 1991, IK+ 77 48 FnNnthc (kparsmcnl 401 Nfilw 11ra1ttr S.icn.ct l/mvtrsiny rol Ilclvnkr, Ikismki V1

C'(1R(1NARY-PR(NIE BEHAVIOR IN ADULT SAME-SEXED MALE TWINS: AN F.PIDEMIOIAGICAL STUDY Ia this attempt to identify frnilid and eaviroruneM.I components of corasry. prone behavior puarsn, the respwsses from 3,119 mak twin pain in the '=innish Twin Cohort ro a 1973 qucsrionnaire were i.vestipsod i. several difkrart wr ys. To beti. with, the postal questionnaire study provided data on zy6osily, anakinR akt*wl use, kisure-time physical activNy, weiRht, heiilN, and drug usaRe. Psyclwjocial facwn such as marital sratus, occupariow a.d oocvpdional hissory, changes of tesideace and empbytneM. estraversion and IabilMy, asd lype A behavior were also sudied. as well as vassous symplorro and history of diswe. Type A behavior patterw was measured by the ntin6 scak devck+~pcd by Sorl.er. Rewlta of this study showed duM the irwaclass corrclatit+ws were 0.2~1 for tno.oryRotic (MZ) pars and 0.052 for diryllaic (DZ) pws. The heriuhility eNrm.nes were hiRAer i. yorwRcr tlur ira o/der ap Rroups, .nd the pniprwtbn of A-type caKnnhia pirs dso sAowed as age uead. Whi k the popu- tinn of MZ pain among A-type eo.erord../ pairs was Rrealn than anwnil 6 type concordant pain, the difference was twl statistically siRnifkarN. 1. tnis study, an associauow io anee betwee. A-type behavior ptrerw aad rrRisa pectoris ow the Rose quesrian.aMe was found. Maroover, 1he diaconlrM pur analysis preseMed here dowed that tAert were some e.vironrnesxal f.cbn clearly associated with corotury- p.rr behavior. As of saw. A-type behavitx has beew sho...1o be r isdependenl risk factor ftx diffentst sns~wfeurions of carowry heaA disease (CHD). Since some psy- clrrsocW facan, such as nurird Narus rd social clau, which were found tocorrelale with A-type behavior is this study, re keows bbe asaoci.led with CHD in Finland. it seems nrasonabk that the rclartnnahep of A-type beluvior, psychosocial facwrs, and (111) shaYld be investigwcd further Knskcnvuo, M, Kaprio, !, Laaginvainio, H, Romo, M. and Srna, S. (Ronrasolo, I ) le: (kdda,1.., Prisi, P and Nance, W E(eds ): Twin resrnrcA ): rpidtrnioloaical nnicliwic.l sndin, New Yucf: Alan R. Iiss, Inc.. 1981. pp 179-115. From the Department of Public Hcahh Science. Univenily of Helsinki, lklsinki. FINNISII TWINS REARED APART: PRELlMINARY CHARACTERIZATION (W REARING ENVIRONMENT This paper presents some characteristics of the rearingeevironmenl of 17t Fin- aish-speakint. aduh, like-sesed twin pain raised apart from the age of 10 or less. The Finnish Twin Cohorl Study provided rhe raw dMa base for this study, and twinship was con/imrcd by a queaiownaire study i. 1973 that eovered heahh-relatcd items ud strndardized measures of rnorbid'r1y.1n addilion to these questions, a number of other aspects were consideced: whether the twin pair lived together and, if twr, at what age sepanlion had occurred. The presear froqrency of intrapair contad, binh order, and handedness were also investitarod ard queslions directed to zyRosily assessmenl were included. Later, during November 1979-lanuary 1950, a new questionnaire on their childhood envinmmeM went out 1o the 478 twin pain in the test gnwp and lhrx correyrrding conuol groups, which were formed to assess which aspects of the rearing environmen/, persanahly fsccMxs, ard childhood medical htsMxy of the study sample deHered 1nom those o( twins of the same age and sea. l.isrings fiM thu study re 6ivcn by a6e o( seprMion (gnxwps 1-1 V) and by bisdt year and sea. ResuMs o( rhis study sM,wed tAa1 the inlnpair correlation of rearing eeviranmerN varied RreNly as it ap- peared from variabks such as age tl sepratio., family memhers, school, frie.ds, h.inR place, inrrapair contact frequency, and educarion and occupation of rearing parents. Morcover, the cause of seprMion, based on self-report, seemed b be fairly oflen associated with some psychosocial pathology. The sepration of nsembers of a twin pair may also mean intrapir selection. Further assesarneal and comparisons with singletons from the general popylation sad with psychiauic outpatients re ongoing for this study. Lan6i..aanio. II. er at. (Rowrnsnlo, l.) In: Godda, L., Prisi, P. a.d N.nce, W. E. (eds.): Twin rtuarcA 3: intelligtwct, Ornonoliry. mdderrfoprwrwr. New York: Alr R. Liss, Inc., 1951, pp. 1R9-19t. F.ooa the Departmerwa of Public Health Science and Psychiarry. University of Iklaiwki, Helsinki. CANCER IN ADULT SAME-SEXED TWINS: A HISTORICAL COHORT STUDY 1o this attempt so i.vestipre the feasibility of utilkiaR the twin method as a ease- eowo/ type of nudy. a historical record-IinkaRe eohort study betwon the Firtish Twio Cohort Study and the Firteish Cancer Registry was canried ou1. The Finnish Twin Cohort wu crered in 1974 from the canpleriaed Ceatral Population ReRistry, while the Finnish Cancer Registry is a populatioo-basod, national registry in operatiow sinee 1953. N is considered a be rather eoropk/e with respect to iscidea cases of ca.cer i. Fi.fand. For this srudy persons included in both registries were identified by com- paring by computer the personal ideulifKMiorr swsnbess (a K)-digN unique code as- signed to each reside.r in Finland) of die two see gi.lries. The cornparirorr covered die ycan 1967-1976. ltrc Iwi. record linkate yielded the observed aurnben of cancers of difkant types. Age and sea-specifie pcrson-yeas at risk were cakulned sep.rarely fa rhe twin pipularion rd the sinalerow Rrorp. Also, person-yean at risk were calculated for cotwins of tancer poba.ds. Results of this study showed that the lord cancer morbidity in the twin population was lower IAr e.peckd for both mca and wamen. The relMive risk for all eancen was 0.77 for aten awd 0.72 fa womea. 1n the singleton ppulalion, the relative risk for men was slightly over unity. Us this srudy, the ratio of the observed to ezpecud cancer rrwrbidity closely rdlected the srandrdized (lotal/ mortality ratios for the same calendar yean. suggesting tha the bwer-than- espetted cancer nwxbidity may have a background in conwnoa wirh the lower-dua.- espected rnoAality. Alscs, the low concordance rNe fornd in this study suggests thN it may be frritful to study the environmenul esposun: of csncer-discurdanr MZ rd DZ Iwins. ` Kaprio, l. tr al. (Ranamlu, l.) In: Gnh1a, L.. Parisi, P. and Nance, W. E. (cds,/: Twfn reuor.-A!: cPidnnioloRkd m.d rlinit-ul studies. New York: Alan R. Liss, Inc., 1981. pp. 217-221. From the Departmenr of Public Health Science. llnivenity of Iklsinki, and Fia.ish Cancer Registry. Iklsmki. 93 92

SLIiIiP 1)ISORf)ERS IN RIi1.AT1ON TO C'ORONARY IIF:ART DISGA':G /:vuknce fnwn an Ankrecan study indicating a rclalinnship between slcep Umc anJ rtrntaldy, inclwhn6 Jcalh (nxn coronary heart disease (CUD). led lu a Fmmsh Twrn ('rdwN1 sludy nn the relali.mship belweca skep time and ('l1D In the cpi.kmio- MsArcal paper presented here, the sleeping lime distributions in the U S A taken fran 1he Kriple study were compareJ 1odalaobaincd in 1975 fnwn 5.419 Finnish adult mcn, and age slandardrzed pu>ptntions were compwed for mcn aged 30 and over. Resulls showed. first of dl, that the pn>p<xlion of inea sleeping 9-R) h,wrs or m.ire is higher in Finland than in Ihe llmteJ States. They also showed that sh.xt (less than si a hours) ar Awrd (rmxe than 101wwrs) skepen had siRlw6earMly tnae complaints relati ns Ns CIID than IMrse who skpl fur seves-eigM bolusper aigM. Shortened skep wa•. especially rrlsted w angina pect,+ris and pain of possible ildaretion, and correlated .ilh asin6, p.ww skcp quality andlor Type A behavior psllerw score. Long skep, cn the rNhcr harwl, was co.relaled with good suajeetive sleep qualNy, but this Rroup had the highest incklence of diapwmcd myocardial i.frclion. This relationship held sUer statislically cantr.rllrnR far many ppwibk confouwders such as hypertension, drug and rkr.lwsl use, smuk/n6 and Type A beh.vinr pattern. In additioe. the cardiovascular physinMsAy and pathnphysiuk>Ry of sleep is reviewed here and the ItlMionship of snmme s/ecrfic sleep disorders to C/ID is discussed. Paninen. M. er al (Rmrraw/o, 1.1 At ra AlrJn a Srandinastira (Supp1) tib0 b9 83. 1992. (hArr srOp.rr: Finnish Foundalinn for Cardiovascular Reseuch. Frn.m the 1)epanmcnts r.1 Neurology. Physiology and Public Health Science. lJniver- si/y of Ilclsinki, Ilclsmki MIq.TIVAR1ATl: 1(X]IT ANALYSIS OF CONCORDANCE RATIOS FOR ()lIA1.1TATIVF IRAITS IN 7WIN STUDIES This stalistical paper presenls a new approach for the analysis of a certain type of twin data The nMKkl that is applied in this paper, the logit mndel, is anahrys.ws to the widely used los-lrnear rtrrkl for contingency table analysis. This trNdcl, which per- rrwts testing of inleracliun effects befare eslinwing the main effecls of the study variabks. may also be easily estetded to four-way or even more cnrnpler, tabks, though the IestinR procedures and sequential brpnlhesis testing becomes increasingly dcmawding. In the esampk used for this study, dMs frorn a cross-nslinnal study of cigarette smokis amunK aduM twin pairs in two countries was vscd. The muhivariale assessmenl of tenctie factors in relation lo other factors was carried out by Ms6i1 analysis of cs>r+crwJance ralios by analyzing three variaMes.:yRosily, ses, and crwe- 1ry. at the samc Irnre Thus, the effect of sea and country on ryR.>`ity in the smolint uail could be controlled. Fur cigarette smnkinR, the present results indicate that in IrMh cuunuies the :ypu.ity effect is sitnificanl, and indepemknt ul country and ses AI- tMwah mN preseMed here,lhe resuhs held far both current anJ ever cigarette smoking as well as fur smoking over one pack a day either curremly or ever. 1•ur the heavy snwiRers, the zy`.sily effect had a si6ni1kaal inleracliun with sea. A siAnifia•ant ryKus- ity effect implies greater concnrdanee fnr the trait amnns MZ than DZ paus and is due hs the akrnlical Aennme and probably greater eomnwin envKSmmenl uf MZ twin pain. Kaprw, 1- Sam.. S, and Koskenvuo, M(Runmsulo. I 1 A, ru (:rnrru,.r rr MrJuur GnNrll.J..gtnr 110 267- 271- 19K) 1 i~.rn ~fr Isrlwv.. m.d 1'uhlw lic.lth Sa renre, lln.vcr..ty ul 11cls.nti 1lrhurAi Active Projects Following is a list ol the principal investi6ators, or in.titutiotn, of projects under way or activated in the period since the previous Report, together with the respective project titles. Completed profeets are listed in a later section. PRINCIPAL INV[!'flCATOR OR IN1i7TUi10N JOHN l. AI BFRS, Pw.D., Rrserch Ar- arN•wrr rru/r.ar u/ MrJiriNr, llaiver- air/ at Washington School of MeJiciae, Seattle. PROJECT TITIi lligb density lipoprolein quaMilaliow HARRY N. ANTONtAUFS- Pn.D, tar.. /rsuw u/ Ri.N Ihr.Nirrrr, /lu.ard Unl- versiry ScMrul of Public Heailb- Boslonn. TI/OMAS M. AUNE. PhrD., AdiaNa iw IrNrnNa.dury. The Jewish Huspita) of St. I .Wis. a BF.RNARD M. BABIOR, M.D., PN.D., rrolrssw u/ MeJiaNr, New Fngland Medical Cenler Ilo.pilal, Boston. LESLIE BAER, M.D., Arwriarr 1"ro/n- aw o/ Mrdicinr, Columbia Uaivenily College of Pbysicians I< SurAeo.ti New York- Humsn platek/-derived growth /ae,r ( POOF ) : relationship to bwwaa albnotderoa: Interferon-activation of suppressw T cell palbways Studies on Ihe mechanism of activation of she tespiratury bursl in neutropbils cigarette snakirtB in Borrnoteasive s./ Aypertewsiva wb+ecu: blood preswn. reaut, .fdoYerowa anA calecbolandr respowsea BEA /. van den BFRO. MD. Research Pulnwwary fu.ctiun Iena in sdolesotan Prdiatrici.n. Ad/anrr rre/raur ia lf/s. aad their patenls-u epidewloleAk arrisriri. Uaiversity of Califorgia apprnach School of Public Ileallb, O.kla.d. RICIIARD l. BINO. M.D.. ho%swr o/ Chules/erol inhibilinn snd cartwm rwo.ea. MrJw inr (rN.rrirwsl, Universit~ of ide ad atberosckrusis SuWhera California School of Medicine, (.os Angeles; Yimrag Asuwiarr is NNa t.iporroleins and the arterlsl wall aMJN'a/ ENX/Nrrr/aR, Californis lanlil- ate uf Tecbnul"y; Awe. i.r ../ Cardi.d- .wr aaJ lnnarNara! Alydiwiar- IlunlinR- ton Memorial /lospital, Pasaden., CA. DFBAIIT K. BISWAS, Pss D., D Sc.. F.Recls of nicwine and bewaoq./pyrene on i l A r ruN arr r../r...n u/ f ra/ R..d.ryrv. hurmone pruJuc/ion I ahuraloty of PbarmacvAuWy, Itarvard School of Ikntal Medicure, B.n1uw. • IRA B. BLA('K, M D., rru/r.av and Nicwine and ncuronal developmear ('lur/- Dn•hinN ../ Irrrrfapu.rnral Nra- rulury, Cotnell University Medical Cul- kse, New Ywk. PIIYI11% B Bt AIR, Pn D, Pra/rnuw u/ Rqulariunof naturat kdler cell activity INII/IMN.Jurr, llnrversny of Caldawnia. Berkeley. 94 95

...i .~~.w rRINCIPAL INV[Si1CATOR OR INSflT\lilON I. MARK BRAI/01/1 ER, PN D, Ac.n- t.wt fro/riror u/ PAarncetology, Northeulern Ohio Univeuicies College of Medicine. Ruocslowa. EDWARD BRFSNICK. Pa D, rrnfesiur and ('Aeirn..w. Drprtmrnt of /b- rhrmi.ny, l1K linivnailr of Vetrwo.l College of Medccrne, Burlinpon. REBFCCA BRYSON, Pn D., Aar.cinr ho/rswr u/ r.~r6.J.rf1. Saw DieFo State Umversily, Saw Diego. CA. VINCFN7O BI/()NASStSI. M D, A.so- ri.er Rrrr.reA R.oloeirr. The Univer- ul~ of C.li/orrua al San Dcep,, la lolla DAVID I.. BUSIIF.E, PdD, Dberror, (:rwrnrr (-rntrr, Aruwr.re rr..lruor of S.ol..eirel S..tn.er, North Teeas Su/e lJnivenily. Drwron. FDWARD 1. CAMPBFI l.. M D, Ar- rnrawt rru/tunr nI A(rJn.nr, Waahin~• lon UncverWy School of MedKine. S1. t ou1s. W111.1AM A. CARTFR, M D. I.o/rsrw c1I Hen1.e.d.sF ..nJ AlrJcr.l Owrol..fl. Ilahnemanw MeJieal College. Phrla- delphia. AI.BERTCASTRn, PN D, Director. Hor- ww.nr RrarnrA L.Aor.rwy: rro/nror of PrrAol..~y end Mtdreiwr. Univa.il of Miami School of Medicine, Miam~ F 1.. FRANCIS C. CHAO. M D., PN.D., Srw- for IwrcNlg.tor, Cenlet for Blood Re- seuch. BoNo.. . /AN F (-H11'ROWSKI. Pul). Aui.wnr Pr.r/r...w n/ Nu.- l.rnn.fry. MeJKal ('ul kge of Virginia. Richmwwl. ('IIRT I fIVIN. M D.. A.armwt rn.fr.- PROIlCI' T171Z The alleraliun of guanylale cyclase by nitric oade eep.ession of cytochrome P450c lrlleractive eQects of nicoline, testosletone snd eYradiol on wciaM change food eunsumpiow and aclivily of male and fen.ale ra/s under 64% awd low ~.o• lein diets Hepran sulfate proleodlycaws and blood AaweoMalre mechanivns PRINCIrAL INVfST1CATOR OR IN11TRli10N BERNI('E 11. COI/EN, Pr. D., P.../esu.. .nJ Drrrc rur, ff unww Grwrrw s/Grnrtn f pJrmwd..Kr Prurram. Tbe lohns Ilop- lins Univcrsily School of HySune and Public Heahh, Baltimore. ROBFRT W. COI MAN. M D., Pru/ru.w of Mrdiciwr, Temple Urliversi/y Schoul of Medicine. Philadelphia. GIDON ('2APSK1. M Sc., PH D., F.u/rs- NM of f•AJlirwl CIKn1IlIr), lbe Hebrew Universily, Jerusalem. Israel. ROBFRT E('11T, PnD , Iro/ruor of Awe- tu.nrt Michigan SIMe Universily. Fas1 1 anuns. 1PROJ[CT 11TR.t Gene1K<pirlneiolohic cbaracleriaia of smolets and nonsmokers IniNiation of plauna coagulation and ti.l. formins syslews in m. On the losicily of o.ygen and auperoaies ion: is wperoaide lo.ief The eRecls of ttyPoaic ttyro.ia carbos nNrwoaide .nd Ire.ImeMS Wueacly hyPotia loaicNy ow endocri.e-liks orlls in airways of young rabbils .nd t.NN leluae CARI.TON K. ERICKSON. Pw.D.. lro- elood•beaiw wmnAoring of suslai.ed.ko• /nwr of TA.rn..rolc.tr, The U.ivetwy tine levels ira tals The bioehemieal end physiological ehr- of Te.as College of ftarnwcy, Auyin. acleriarics of a prorerw .hich specifl- i caey binds polycyclic aro+oalic hydro- V. OFNE F:kWIN', h1.D., rru/ruur of Actions of wicoline on isolated Perfueed carbow. Ilcwrn.rulugy: Dr.w, Universily of wour brain (:olor.do Sebd of Pt.erm.cy. Boulder. Modulators of inllammalo.y cell proleoly- Efeeu of doolble orl weuropepida sarse- tic aaivily /iow by irr.n oaouse Mnn, a Fh.en.ee• pewnie rudy ALVAN R. FEINSTEIN, M.D.. Pro/niur ' SnmLiry. detection bias aad MYwary h" The in/erptay of (mmumnurveiilance and of Afrdic iwr A t piJrwd.dull~~ Yale Uni- e..eer imerferun inducliun in Wrnwilenesis versily School of Medicine. New Haven. CT. JOSEPH D. FELDMAN M l.wruua.. D E aRir awd li id n th iomu.. . ., . p.tAul..rlsr, Scripp Clinic and Rese.rch ;~~ y p , o e Nicotine in blood: detection by radio- Fouwdaion. L.IoN., CA. ienmunoassay THOMAS H. FINI.AY, PI/.D.. A..arirrr Structure. ProFer/ies awd ee4ulMioo .e rru/ruor of (tAqruirs and Gywrcoludy, .aolase plaura. praease i.hibNon New York University Medical Center. New York. Platelet activation and blood hypercoagu- labilily BIK(i1TTA FLODERUS-MYRRIIED. t?PidauioloRie research on the SwedW hr.D.. Ar.btunt rro/eswr of Ent:ruw- lwiw registries mrnt.l N1~itnr The Karolirnla (lowi- ( IWe Sloc\hOln1 falufimelric invesligaM.n uf proleinau-.y , . macroglubuhn imcracriun 1l1D1t11 ANN FOSIER. PND., Prnfes• lnvolvemenl of elauin in lung disear wr und CAauyrrs..n. Drrorrncrnt o/ fladury, Sycracuse Univerwy. Syracbse, Biochemi-Ary and function of In.man /ran- NY. ~ wr ..I ()w..J..Ky • Prdwracr. lhe Inhns Ilopkins Oncology Center. B.lu• ulopDn'Iw: anllgens lT N m m J .P tV A mole. ('IIAR11 t() ( IK 1/11ANF, M D, A(rw.- brr, Irr/.rr~wrwr ../ Iwcwcrwa/..cAolofl. Scr.pp~ ( l.wu •wd Rrse•rch Found• Iu.n 1 a/ull• 1 A 6 eJ.Nion s)uems in inflam.natory lung disease /A('K W. FRANKFI, Pit D.. Cun.wOrnnt in AftJwul Rr.ewrcb, Velerans Adminis- 1rNiun MeJical Cen/er, Ray Pines. FL. ALLAN P. FRFBDMAN, M D., Aui.- une f IofraM.r of Mrd..inr, Ilahne- mann Medical Colletie, Philadelphra. Smoking and lung cancer: dia`noMie IeM to identify Pet.ons al hidh tisl The eAec/ of ciprelu neoliq e. Iha dveolar ekaramco rale of inerl duu Parlicles in the hwnan luly 97

PRINCIFAI. INV!',T1CA TOR OR. INS11 rUT1ON AAR(1N 1- FRFI MAN. Pn 1). %,np .C, i- rnh\r. ('JI/I,Mrwa Rw.nsrJ.:al Rcscuah 1,w,nJah,.n. I a l.dra. ( A KiFI 1 FUX1. &I1).Pr.-fr...w../ Ni.nd- .-.r, Ihe Karul.nsf,a Instawe. Saicl- Mdm. M(1RIl)N QAI DY1ON. M D. Aswrfrrr Pru/r.sur ../ AIrJw rnr. New YaWI. Uni- versity MeJ,cai ('en/tr. New YurL MtC11A1'1 C. OFOKAS. h1D. PND, Prn/rrrw .n/ 1'.rr1'Aawwr.n. Dr~..r- nrrnr of Aftd.clnr. llniversily of ('a11- lornia School of A/edicrne, Davis. TERESA OESSNFR. PrrD. Auocl.rr Canrrr Rrrrr\h Srirnrrrr, IIeaN! Re- starch. Inc., Rwwe11 Par\ IAvnwn. Ru/lalo )M-Ot1FS E. GIEI FN, Prr D. Ars,ri.rr P.nfrran, cae...arury of AlrJrrat ('benrurry. Tu.wnl..p an/ /lyeane. Institute of PaholoAy. Univers\ty of I r2lie. I Kgt. Rcl(.unl. GARRII ` C. GO/)f•tAN. M D. P..•f. r s.N .r/ P.dlr.,l.-K.. ('.JumMa I/n,srrsay ('ulkge u( PAys...ans & 1urgcuns. New Yurk . WARRI N M. GO1 D. M D. Pr../rtsur of Afrlni,rr, CarJwvaaculat Rescuc• Inuitute, Univeruly of Cdifornia, S.n Franciaco. SIDNEY G01 DFItif11FR, M 1). Pru/rs- sur u/ Purh..lory. Alberl E:inslein Cof- kAe of Mtdicrne. l he Ilrona, NY. MAURII-E GRFFN. M 0. Dnrrwe, tn- snrrrr /.. M,dn uf..r Yu.ducy. SI. I ouls University Medical Center. St. Louis. MARK 1. GKI'FNF, Phrl), Ass.wiorr Pr../rswr ../ Parh.dugy, Ilarvard Med- ical Sthoul, Ruston. IIIRA 1(i1/RTOt). D V M.. 1•1 V. Sc. h Prr D, As\w wrr ('..n. rr Rruwh Surnuu. nrrwonnrnr of !:\prnrnrnmf Thr.arrrrn,, Rns.ell P.r\ Mrm.uul In\u~urr, Huflalu PROJECr TI1LR Rodent anJ human lung epirMlial cell cul- lute as a tool fnf carcNlO/enesls reKarch in vuro FpNhclial cell cucimycnesis in rdr.. PRINCIPAL INVEJflGATOR OR INSflTi1T1ON NOI/I/Y()111I IIAGINO. M D.. Prr.D. Pn./r\s..r ,./ Anah.rny. l)nivcr+ily 4.1 'rcaas IIca1M Sc.cnce Center. San An- /unru. PRO/RCT 71TLE Nicutinic recepuu, of I I/R11 acun lera- inals in Ihe meJian eminence CAROI INF R. IIAI 1, M D. As.ori.ut Inlerrclariun.hip uf infectiwms lower res- NicWine. c.arcM.lamines anJ nruruenJu- Pru/rrs.w .4 PrJwrrio s and Mrdw inr, ..ry Iras1 Ji.casc in in/ancy, anJ enne(unctwns University of Ruchesler School of MeJ- nt and envir.uwucmiil /acl.a% to later t i.ine, Kuchcsler. NY. dcvclupmcM ut chrunic IunA Jiuase Rioehewtical basis of prrJnpusNa\n to I INDA M IIA11 . Psi 1). A.an inrr Pr..- Genctic dillerences in nicotine sensitivity elorunic ubslruclrve pu/n\unary disease /ruw .•1 Grnrri. s a/rJ NIMruN Nm r. in Drowp/rifs wrrlrnurasrrr slrains Albeit Finuein College of Medicine uf Yeshiva UnivcrsNy, (he Rrum, NY. Crrculating pancreatie elauasr 2 anJ em- phyrcnra in man P\.rsacoRenNin of eow4uAations and Iury eancer risk MuJulaliun of atr/ byJrocarh.m hydruay- Iax and epuarJt hydralase in animal tissuts anJ in telt cuhure hy cigarette smu\e eumknste anJ urher shemicala Riocsemicd mechanism(s) end qualita- tive and Quanoiutive urosrauences of hentula/p)rene mtlafrd..,n ( ylu.leltul .nganu./NM u( Ihe enJu /hel\al \tll in trgULln.n u/ .h.pe aun IracldHy rnJ surfaae muvrnanl Fllecl of utune on airway mau cells Eatraeellular matria-eylochemistry and rhrasrruclure Amplification of human aJtnwirus trans- fornrN/o11 ploltlns in prularyulK and cuLaryaic cells Suppressw cells in synjencis tunwr im- munity Rule of genetics and pulyarurnaic hydro eatlMrn melahulnm in humnn lung can- cer 98 t PAUI. IIAM(rill, M 1)., Ars...larr Pro- /rsr..r of PAysiology anJ Bwphy.hr, .nd AfrJrarnt, Geu.getown Universiry Schuols of MeJicine and DeMislry, Washington. D.C. NORMAN W. IIFIMS'fRA, 1'r1D. Pru- /tsrw of Pa)rAu(ogy; DVrcr,M, Nrnwn FKI/Nl Lab.rat.ny, U.iversity of Sowh Dal.ula, Vernl/tlwn. HERMERT ff. 11ERSCOWI f7, Pe D.. Aswaiart Prulrator o/ AMicroNufugy. Gcorlieruwn University Sdrrols of Med- leina and Demis(ry. Wasleinroq D.C. RORFRT M. 1101 FMAN. Pu.D., As- ' sraranl Pra/.•suw of PrJiMric'c in Rrsi- Jrnrr. Univrrswr uf California Sclwol .d MeJicine, L. /utla. 11=KOM1• 1. II(/INAt'Kl. Pu.D. As\is- rnnr /'n./r\an .a/ NnobqN.NI J,rrmr\. Univer»ty u/ I uwcll, I uweR. MA. WAYNIi 11(ri1. Pn.D.. AIr.M'irrre Pru- /rs\.w, ('enler f.w Srain Research. University of R..ahester Medical Cen- ter, Ruchester, NY. AARON JANOFF. Prt.D.. Pro/rrr.or of Pathuln[y. /ledlh Sciences ('t:mer, State University of New York at Slony sruuit. Stony Bruott. M(IRR1S / KARN(1V1/:Y. h111 . R (lr.. \h.nnn l Pr../r uw ../ rurb..h.F n wl.Iho- r.nny, IlarvarJ MeJa:al Schuul. Ilsnlun. INGY(iAKD M KI:I IH. Pn 1). A\si\rrnr PL./1'\4M ,./ Annr,nnr. IlmYCysrty Uf Wixunsin Sclwwd af Velerin.ry MeJi- cinc. A1aJrwn ('i faretle ..nuttt and Iipupr.Nrin remodel- rnee by Me hsnle Sume heYaviotal aspecls of unotiq and srwohinll deprivaliun Fffecns of cigareue \nwle espusare on de- velupmenlal. celhJar anJ rnuleeular n- pecb u/ Ihe nurounc respnme Methiunine Jependen.e, nselhylation uld urfanlc IransfWmalNM Regula/iun uf .eQalar unc\ycnes Nicoline-inJuced changes in primde high dcnsily Irpuprutcins Sludits of nicu/ine interaction with bloal eells Furlher uuJie. .ur suppressiun of pro- kase inhibition by cigarette smo\. Immnmdoelic assay uf lung elarin de- {r.JNiun The nwkcular haas u1 ppdmunary swfac- IaM fKreliun by 1)pe 11 pntnmucytes: studies in intact cells anJ a ceR /rte sysles Part 1: Lung neuruenJucrine cell innerva- liun Par1 II: -1ran.plxemal ctlccr \d smolinA on lung n[ur/KnJtKrlne ctlls in Ihe n.unale 9/1

PRINCIPAL INVGSi1GATOR OR INSIlTUT1ON ROBFRT W. KKFII ICK, Prt D., rru/rs- ror of ('hrmntry. Untversity of Roches- ter, Rochesler, NY. KI AIIS E. KUFTfNI-R, PhM D., r-nlrs- wr &wd (-hrirn.an; Drpwrnirnr0/ {o• rhrmnrry, Rush Collest of Heah~ Sci- tacts and Rush Medual Cdkge Rrsf•- Presbylerian-St. I.ule's Medical CeMer, Cbta•o. I.AWRFNCE I.. KUPPNR, PN.D., Asso- rwrr rro/rrr.» of !Nn-aiuw Uaivtr- sNy of Norlh ('aroliaa Scf~ .1 PM- lie Heahh. Chapel 11d1. ABPL I.AITNA, Plr D., DrrcMw, Now York Slate Research ImlilYle for Ner- rochemnuy and DrYR Addtction, New York. DON !_APPNAS, M D. Auirunt rre/es- ror of roarhoG.cy. Ilweverswy of Ver- wsorw Cdkte of Meduine, Bwlin/low. F C11NfON l_AWRFNCF, M D. Au4- rtant rro/rnor n/ NrdN inc. Baylot Col- k/e of Medicine. Houuon. P11111P M. LE VUFSNF, PND, DSc. rro/rtusr of CArmnrry, Nurtkauern Umversily, boslon. FABIAN 1. 1 IONFTTI, PN D. Rrirrrh rro/ruw of Slachrmiury, Boston Uai- sersity School of Medicine. Boslon. GESINA L. IONGFNF.CKER, Pw.D.. Asroriue rro/rssor of rharnucoloRy. Universily of South Alabama College of Medicine. Mobile. RONALD B. I.UFT10, Pu.D., rru/ruor, oOf t•frToKology and lmmanofo~y, Uta4 verlNy of South Carolina $iod of Medicine. Cdumbia. HENRY T. I YNCH, M D, rmlrswr onf CAaYnwn. OrNrrnsrnt of hrrra- rlre h(rdKinr and rYNk Hrdrh, Cleilhlow UniversNy School of Mcdi- eine, (Anara. PRO1tCr 11TLR (nvesli/alions of the imcraction of nico- llne with rllelnbranes Local regulalion of normal and Padbololoic invasiow RegulaNion of Proliferuion of invasive eena Verificatioa of a statistical age-period- eohort andysia of lung cancer PRINCIPAL INVfIaflGATOR OR INSIITUTION 1. WISTFR MFI(iti, M D., (Y8nirnl rnr- /r,to- of Epidrmi.d..xor; f/NtIhM, (-un- nn ticrt Canrrr #I.'(~ rnri..I.rRr Utur, Yak l) nivcrsily Schoul of Medicine. New Haven, CT. FF.RID MIIRAD, M D., he D., rr../nu.r of hlydarne and rhurmrruh.Ry. Slan- f.wd Uwivcrsily, and Chief of Mrdn inr, Palo Alto V.A. 11ospilal, Stanford. CA. IAY A. NADF.1., M D., rroJesMN of I+Irdkinr, rhysi.drKy and Radldogy, Cardiovascular Rerearch Iwslilwe, Uai- versily of California. San Francisco. PROI[cT -nTLfd Review of lung cancer in Conneclicuw, 11)3 presenl cchaniue of wiaric otide adivalion of louanylale cyclase Rolc of cyclic GMP in smooth muscle rclasaliuw Mcchasusms of drway ItyPerirriabilpy OenNic basis for nicoline responsa D(1NA11) 1. NI:1 S()N, Pu.D., Aswn(atr ' SpeclruscuPic uudies of the imeraclioa of rrU/rJMM of ( hrndstry, (larl Univer- chulinerolie lipnds with nkwinic teeep- sily, Worcerer, MA. lor poleiwa The association of inorpnic dust defqsi- IIAR(N 1) H. NF.WBAI 1., M D., AsNMi• The rok of prweases and anlipokafn in liow with pulmonary neoPlasia in /o- rfr rro/ras.r, nl Medwrinr, 'fhe lohns pulmonary emphysema bacco users 11uplins University School of Medicine, Bahimore. EAec1s of ciRaeue smo-inR on immuno- lobul~n poduclion by human bronchial FRANZ OESCH, Pu.D, rro/ruor of Metabolic /ale and loticological si{niR- ymphocylcs I rhra•nmacoloty: Nral. Secriwr on /lo- caace of dihydrud'wls derived from chrmicd rhrnrcoto~y, University of Ddycyclic aromatic hydrocarbona oc- Asuy of oayRenated suerols in human Mait~ Main; Wes1 Oumawy, ewting in cigarette smo\e blood vessels-a feasibility study Pha/ocyte mediated injury to lissues BF:NDIC111' 11. PAU11. D.V.M., A»..ri- arr rr../rss..r of rahofot , Rush Pres- bylerian Sr. l.u\es Medica~Cewler, Chi- cato. Iucal re`ulaliMl of lumtx ineasioo by hoal+krived prweinase inhibitors Studies of plalekl and endwhelial Prosla- woid produclion as possible cardiovaa- cvlar ris\ itadicalors is smuliers Interacliolls between RNA lumor viruses and chemical earelrwliens (knclie and biomatier audies of smo-- in/ rssocialed cancera BRU('F. A. MAl'IIFR, Pu l), Anittant Chemistry and biology of comples earbo- rrolrrwr of rh.rm.<rrtaaf Chrmiury, hydrates Ilmversily of California. San I raneiseo. Al AN (' M41 Al ll;lll IN. Pn 1) , hrn. p1. r ~.. nr N.,..I M•rn N.i ,..nal I at.Na !.w r t 4ww N \ Interaction of divalent csfnms with model and Fadupn sl mrml.raIes DENNIS R. PETERSEN. Psr.D.. Assisr4u rr../ru.n of rhrnwr.d.y!r, Uraversily of ('ulwado School of Pharwsacy, Boulder. 1U11A M. P(N AK, DSe., M.D.. Seniw f rtrwrrr IN Ihst.qrnlwh.r , Royd Posl- {raduale Medical School. ~/amwlerswtilh Ilosplal, I.oabw. W1111AM A. PRYOR. Pu D., e..yd rr.. /rsMw of ('hemistry, Louisiana State Ilniversily, Balun Rouge. 11 ARI RANTASAI 0. M D., rie/rs-or rnd Chawnran. Dr tmrwr al rrhlir Nedth Snrnrr, U versily of klsiatai, Ilelsin\i, Finland. RONAI D F. RASMUSSNN, Prr D, Ar- iorbtr AlJuwrr rro/rnw in Conuua- nNr and Enrirowmtnrd MtdwJnr, Ilni• vetsil of California College of Mtdi tine, rriwe. 104lences of Cenoqpe, sea and chroak cigarette snwtbinR on nicwiwe and al- eohol metabolism is rnia Inveslifaliuw .4 The ruk of re/ulalory /eprdes in human lung disease Fret radical chemisuy of cigarette sasole The Finnish 'Iwin Coborl Folbw-uP Study The rult of « R-spccific lusins In nsouse htnt catcinoWnesis

PRIN('IP A/. IN V F:ST1G A TOR oR INSTIT7ITTON F1/ 1'1'N kf MOI 1) -O'1)ONNFI 1., r11D, I't.n.IpJ Nr.r.r.h Ass..wrt, (1a/.arJ McJlcal \aMwJ, In.rulcrlar, ('emer /.w Slood kesearch, Ilouon. /OIIN F RI PINI'. M D., A.siuan/ DJ- rr.l.M, Il'rl.A•WqrIN'- I Mn[ InVItWr; A aaw LNr lr..lrss.- a/ MrJN inr, Uol- versily of Colorado Health Scienees Center. Denver. IIFRSERT Y. kFYN()t DS, M D., Pru- frssnr .f AfeJN/Nr; JIrN1, IN/nwNrr Srrri..N, Yak llniversi/ School of MtJic.ne, New Ilaven, C1Y. PROIECT Tln.a PuriRcalion and funclional analysis of elaslase from suinea pit macr.phates Sasic nscchanisms of lung injury from in- hakd ouJanls ReyirNOry secrclions In pulmonary cw- eisowrr secrelory component ol in- m-A as an early alarker o( ePil~ysfutsctiow PROJ!(T 7T1 Lk Sludies of macruph.ge sutporulations and diRercnlialion using monoclonal aMit/odKs l rans/er of ePecilic inJiviJual human eMumoswnes 10 recipiem eells Arterial degradation of low dcnlily lioo- pr.Meins in rrru Functional :nqomy of the lung macro- phaK PRINCIPAL INVEITIGATOR OR 7NSflTU71ON TIMUf11Y A. SPRIN(:FR PwD., As- alu.nt fruJtrs.w of raJiulury. Har- vard Medical School. Sotlon. I:kl(' /. SI'ANBRIlX:F, Pu l)., Assu.i/ur f rufrlNN o/ MN...hud..rr. UniversNy u/ California. Irvine. DANIh.1. SfFINBFR(i, MD.. Po.D, rr../rsuw a/ MrJNinr, NrNJ, I)iri.ioN of AlrtrA..llr Disrrsr, 7 he l/niversily of California at San Diego. La /ulla. THOMAS P. S70SSFL, M D.. Chk/, Mthrd Onroln y UnJt, Mauachnella Oener a1 1/osp1.J Itoslo.. IR(i1N/A 1. RI( 11MON/), ru.D, Rr- sru.. h Aiuw.wrr, Pacl(u: Nurthwest Re- sca/ch I .wndatiun, Seauk. Mrlers of ePithelial cell dysfunclion in respiratory secrelionf of rmokers Elaslie flber microfiMillar protein slrue- twe 1). 1 ANSIN(i TAYI OR, Pu.D.. AsYN lnlr Pru/ess..r 4.1 di.dury, Ilarvard Unirtr- .ily.('aleMN/ge. MA. JAMES TRAVIS. Pu.D. rro/t,aur of 11ocAendanl, T~e ~laversiy of Georgia. At`ene. Pt TI R M ROSS. Pu D, Rr.eurrA Ars.. rlnrr, 1 he Roclelcller Univcrsily, New York. DNA damage and sekcfive maintenance of animat senes EMIL R. UNANUE, M.D, AfdGncAruJt Professor of fN.m Aulop, Har- .ard Medical School,ow. STEPHEN F. VATNER, M D., Aaawlarr UNA S RYAN. Pu D., Rr.rrnA Irn/ts- Intcracuons of hormones with ulls of Lofeasw o/ AIt/iriw. Nar.ard Medi- s.w n/ Afrli.inr, (Iniversi4y of Miami Ihc pulmonary vascular wall cai School. New EstRland ReRio.al School of Medicine. Miami. FL. ~r6n.1e Research Cenler, SuWMroeo. l/A ; Ass•ui.ur in AfrJNiNr Pe1er Rent 9. V. RAMA SASIRY. DSc, h/D, Influence of nicoline on Ihe «kase of , Srigbans Iksspital Suslow. Professor of IAarru.rofn~ y. Vander- acetylchohne is the humanp1acen/a aad , lilt University School of Med/cine, ils Mnplicalions on the felal growth PETlR N. WALSII, h1.D., rru/rssuw o/ Nashvilk.l N. Meli.inr, Teasple Urrversily Schuwd of PhiLdelPhia. Medrcine IS/IAIAHU SCIIFC'IITF.R, PuD., Sewi.r F.Aed of Ihiols and disulfides on dsoks- , Lrrr/nn iN Ii.r hrnu.rrr. The (itor~c S. (erol meuAolisrw OEORQE WEINRAUM, Pw.D., lioarJ- Wise F'acahr for I ife Sciences. Tel Aviv rnrisr. rubnuNw' Diuase St. riuN. AI- l/nivenNy, Tel Aviv. luael. berl Ekatein Medical Ceata, Phila- OERALD SIIKLAR, D D.S, Charlea A. Oral carcinoitenesia, .itami. A and ret- delpilia. 1rarlen Professor of Oral PuAoloAy: iwids r tN.rnr o/ ad MelVirint Dt Ife.f I. S1:RNAkD WFINSTI:IN, M.D.. f...- ~ . and (tid rarholon. Harvard School of /r...w n/ AtrJir iNr rNJ tbs ir..NnwntNl (-uhrmbia Ueiversiey, New J.irn.ra DeMd Medicine. eoston. , Yurk. kO11FkT /. SKI AkFW, Pu D. Rrvr1A Asww/wrr Pr../r.N.r u/ PNthuhryry, New Y.w\ llniversily Research Service. (ialJwaler Menwwial Ilo.plal, Ruox- veh Island. New York. HANOCH St OR. Pht D., Asanrlaa rto- /rrsor n/ NrN/an Grnnits, Sackkr School of Medicine. Tel Aviv Uni- versNy, Tel Aviv, luael. DFNNIS M SMI1l1, hr t) , Al.iuant Yro/r..... .u/ l1/.•I••ra u/ laurn. rs, Wcl /esky ('.dlege• Wellr.ley, MA Chenuwaeisld maerophases Pro(eolylie enzyasea and IaAibUors Is a.- .h).e.. M)'sioPtlbb~p of normal and activated saaYCS~hyes Dlreea e%c(s of nicmine on brala ekew INIom laeraciior u/ Plnekts sril\ coaRWatfow faalyra IX and X RroweNo.lseda lavye of hurnas swot- en ../ Iso..arokers: studies .e oeM che/wal.aie, enaya relaue .d oaMr- Lr ullr.aryd.r. D.weMqwteet of mnwrclunal amiba/ks lo earcinuien-DNA aJJucts Cylolinelicsof heleroploiJ wlpopulations by imaging SIl3MlINI) A. W1:117MAN, M D., As- Sludies of PhaRueyte-irw>,/ceJ mulauiow ar.NUnt h.l/.YMN ../ AfrJa/ne, Ilcnra- Iu1.Ky-(Mculasay IInN, Massachusetls (itnnal (Iuspilal, sustun. • The uae of specific antibodies so wsonilor AKL' WENNMAI.M. M D., A.r.rirtr ' Nic.Mine rs inhibitor of pus/aijtandin for- Ihe formalnrs and removal of hento- Iinnai rhysl.d.rytt ru Ir../rss.r .J ( Iua1NM: h4'alilalMM of the inhib/ory la)pyrene a.Mhrcts frum DNA of dam- IntI1tMIr, Hudd/nte Horpl- step.wd characleriaation of the eardio a{ed human eelh lal, 11udJrnfee. Sweden. vascular irnPlica/iuns Aulausmic cunlrul o/ pdmonary surfac- 1O/1N 1'. Wll S()N, Ptr U.. Aul.t.alt rr.•- lhe hrrlaliun anJ espressiun of hurnaa lan/ in Ihe adulllung /'swr ../ C.'ll uNJ Af.J..wl.w Nl.d..kr. .-1-arwNr)p.iw ernc scyuen:es tlwough MtJ~'+1( •olkge o/ lft.w gu. AuguNa nsukcular cluninte 102

-%I rRIN(:11'AI. IN V F3TICAT(IR (OR INSTITUTI()N ALVIN WINTERS. Iw.D.. Asslstewt Professor of Mrlicet Mirrobrofop. Vderera Adminiuruion Medical Ce.- Ier, Bay Pines. F L. BRUCE A- WODA. M.D.. Msori.tr tro- /ruor oJ raAoloty. Urti.er.Ny of M..- a.cbucetlti Wo.ceaer. STANI.EY YACHNIN, M D.. Professor of Mr/.f rwr ewI C'Air/. Sreriow o/ New.- roloty/Owrololrr. The Uwi.enYy of CbiuAo Me6cd Ce.ur, CUicap. rRO)E(.T TITLE Efecte of .rnolieg oa the irdiereel inter- (erow k.els in coMtol eed cawcer oe- tk.u: a Eilot Mudr CeR surface t+tembrwS iraesr.l reets- bra.e proteiiu .nd eytosit Wow iw lywi- (rotn: (1) youry and dd r.lr, 2) eroa..cow ra (rrwN,>,w.. ModeN for the~r.tM~teoee'e of .uMno- akro.i.: A) biololrc.l dlects of osr• gewMed elerol eorwpwwda; 8) ioev.lonk trid .nd cboleMerol bio ymbesi. .wd tbe biosyybesie .wd relulurow of ceR Vowtb 104 i Completed Projects Polbwio` i. a Ibt oI the princip.l in.eqft.lors, or iauilulioa, d projecb that have been completed prior to the period covered r this Report. Several of the iadividuais named ue deceased. The 8da sd.mll.- lioar lisled aye those io effect at the dme the work was eoopleled. LEO U. AeOOD, M D. Professor oJ IlocAtrntrtrr ewf l.oiw ResrrcA. Cew- /er fur Rsei. ReseucY. ?M Uni.ereily of Rocberler Medical Ceaer, RocAer 1er, NY. MARIO D. ACFTO. rsr D., A,socib. ho rsr /w.avfottr. Medical we.k` U.iVirer y. kb.a.di.ia Cowono.- CLARENCE M. AORESS. M D.. Asse• ciere Cliwicef Professor of dledklwe, Uwi.cnit~r of C.iiiortri. I~Iedicd Ce.- rer, Lo. Ayeke. ANTHONY A. ALDANEfE, ht.D„ DI- rectr e/ l.boeotorks, Tre Drtle Re- A.biliuAioe Cewler, Wbile Mairm NY. ANTHONY P. AMAROSE, ht.D. fw- strrnr /w Oftistrtsks o.d Grwrcdo~r. Tb AR+.or Medlcal Copqe a1 U.iotr Uwi.crsi/y, Albany. NY. E. T. ANOELAKOS, M.D, rr D.. lro- /essor of f•Arsiofotr. Reao. UrtWereil7 Schsol o( Medkir+e, Borar.. D. MURRAY ANO8VIN8, M.D.. U.1- •ereNr o( Wl.ceselu School e( Med/- eise, M.dieo.. JOSEPH C. ARCOS. Dsc.. Professor o/ A(rdktwr. Tut.we Usiveniy School of Medieiee. New Ork.we. ALAN K. ARMITAO~hrD, R.srrcA DMectr, ILslelo. .boraotie. E.- roye. H.rroptk Narth Yerteliitr, P..g- lewd. d(ARILYN S. ARNOTT (RASCO), Ptt.D, A,sutewt /1ologW ewd Ass/stest Professor o/ 1iolosy. T1e Uwi.enitr of Teau Syaere C.ocer Cewrer, M. D. Aw- dersow IIoyN.l .ed Twrtor Im/wWe. Houaow. DOMINOO M. AVIADO. M D.. lro/es- snr o/ thrmerology llwi.eru"y of reeecyW.ei. School oi Mediciee. nil• .delar.. STEPHEN M. AYRFS, M D0. DMecter, Crd1o/rlwrew.rr [A&rerrr. S.k/ ViecetM'e HoyiaL New Yort. OSCAR 1. BALCHUM. h. D., /Ne.Mng, Professor of Me/k/we. U.isaY~ .( SuwAerw CdUwwi. Sclted ef M.M- ci.e. Loe Awgela. FREDERIK •. •ANO, M.D..".u.r owd CA.irwr.w. Der.rrw.ewt of rrAr No(o~~. Tw laMaN~t i.e Uei.enRy ScboW ef HyRiene ar4 Public IIe" R.biwwee. A. CLIFFORD RAROER, MD.. R.6..I N.w.' r/Nser rro/.asor of PAysW- op. Nr.ad Medical ScbA AeM... RRODA A. RARNES. M.D. P.D, !r. /sse. (A/tlWe) of rA/Mole~t, C.M~ nds Sula Uoi.eteMy, F CoRYr.erl FREDERICK W. RARNP.S, l.. MD. Assoc/ert Professor o/ )tledklw.. ib loMn Ilortlr U.lsteiy StMd ef Medicirw, N.Miweee. T. C. MARNPS, D.Se, Rese.rrh Jeia ~ .de1Ml. S(.g. HoqNd, h/.- CARL O. DECKER M.D, As.x1w Professor of lrAo~op, CorMV UoF tereity Melkal Colkst, New YorL. R. FREDERICK RECKER, tiD, Arr dae Professor of Aw.Iowrr owd DbM Ir, l.elorNOrr of rtstwrr Scie.cv. Drle Uwirereity Medical Cc.w. D.r- Aaw, NC. RALPH S. DECKER. ft.D., Professor o/ CAe.e/strr. Uwiverdly of 11wdar. IIo.MoO. BENIAMIN RFL1., M.D.. Direcsor Ewrer- Ars. Nrwrrlre A:4rd Study. Ve/et- w Admi.iMrNio. OwprkM CY.{c. 11owow. SAMUE/. RELL@T, N.D. Dlr.ctr DE r/slow of Crd/ology. lWdr6W Oewcr.l Hoqild, rWedelphie. 1us

BARI/l BENACF.RRAF, M D. Fabr.n rru/rsl.r AndChY1IA1Nn, IIr141Iln1rnr P/ rarhulury. llarvard Med.cal ScAuol- Buslun. WII I lAhl /-. B! N1 D1Cr- M D, Arsir- tant rr../r.r... ../ rrJ..nru.a tlniversity of Soulbcrn Cabf.wnma Sch.wI of Mcdi- crnc. Division of Ilcmalulury anJ McJ- Kal Otnel.cs, Chddren '& Iluspilal of l_os Anreks- I os Anrelcs. JOHN A BEVAN, M D., rr../nsr ./ rhrnraofury, llniversily of Califor- nia School of MeJiciwe, Los Aryeks. BUDIIDI?V BIIA()AT, PM D., rru/rssr of rhlua6.Ly. Saint I oais Universily Scbuol of Med.cine. S1. I oris. CFSARE BIANCIFIORI. M D, Dir/don of Con.er Rrsr.r.h, Universily of Peruria. Peruru- Italy. HYI AN A. BICKERf.IAN, M D, Assist- .nr rru/rswr of ALlninr- and AL- VAN 1.. BARACH. MD, Cuwsu/rMt in AIrlu.ne, Columbia Universuy Col. kge of Physicians i Surgeons; Gdd• water Memorial Ilospilal, New Yat. B1O•RF.SEARCH CONSUI TANTS. IN(', ( ambr.Jefe, AIA. BIU-KESF.ARCII INSTITUIE, INC.. CambriJre, MA. FRED O. BOCK, Pu D., Auoci.te Con- cn Rerra..h Surnriu, flwlorrcal St.- tnon- Roswell Par\ Mernorial Inslilwe, Sprinrville- NY. GUENTHER BODFN. M D, Associorr f.o/enor of Mr/aine; Auutant Dre.- rr, Genrral Cbnira/ Reserch Centrr, Temple Univer.ily Iled(\ Sciences Ce.- ler, M'dadelpAia. HERMAN V. ROENIO. Pw D.. Hrod. Deprtnrent of Che.nistry onJ f)wxhenr- btry, Spindletop Reuarcb Center. Lee- inglon. KY. JAMES F. BONNER, PN D., ProJtssr ol ~w~ry. Caldotnla /nuiwle of TecMolory. Pasade.a. WAI feR M BOOKER. Pw D, Pro/rs- sr owJ flr.J. Urpr tnrrnt of rAunse- eo/..rf. /loward Uwivtreily. Washing- Ion. Ix • FNANr /11\ M reM/Y\I Pah \rn.w /w.r.nr.r... Mwkal Nrrsc Rrvaais &.(.ro RAYMOND BOSSE, Pu.O., Aa..riwrt WIl.1 lAM H. CARNES, M.D.- Uni.er- lU1.Il/S 11 COMROE, /a., M D., Dkec- Unr4 tur, N.nnwnrr AKrnr .SruJr- Ve/- sily of 111a! College of Medicine. Salt tr, CrJu.ruscular Rnrrch lnstlrre, trans Administration Ou/palicnl Clrnic, I.ahe City. University of ('alilorma Medical Ce.- Buwon. ter San Francisco MARCUS N. CARROII, la., PwD., , TOM O. BOWERY. PeD, Rrsrr.h Chief. Diririon of rhrmacaloAy, T11e DEAN M CONNORS. M.D, Msoci.tr rro/rswr. raticree Rnl/ur Labore- Brootdale Huspiral Center. Brooklyn, DMectr, Drpr tment of l.abaratry to.~Norlh Carolina Slate College. NY. Mdicinr. St. M.ry's Ilospilal, Madison. Kakir\. W1. OEOFFREY L. BRINKMAN. M 0. As- aorMtr Professor of Medicine. Wayne Slale Uwiversily School of Medicine, Detroil. ROBflRT E. BROOKS. Pw D, Associue rro/rswr of Potholory. Universily of Oregon Melical Scbol, Portland. BARBARA B. BROWN. Pw D., CMr/, E.rrrlnwntol Pi)chWry, Veterans Ad• waei.(raion HrnpilJ, Sepulve Ja- CA. RAYMOND R. BROWN, Pst.D., rro/es- sw o/ Cbnkd Oncolory, UFiversily of Wiscowsin Medical School. Aladiso.. JOSEF BROZEK. PN D. rrolessr awI CAairn..n. Dep+rrnrrnr of Psychology. I eAirY Universi(y, Be(Idebere, PA. I WILLIAM ALVIN CARTER. M.D.. Assistant rru/rnsor of Mrdirine .nd Afnrobwlutr. Tbe IoWn Hoins Uni- versity Scho.A of Medicinc, allirnore. I F:OPOI 0 R. CERECI:DO, Pn D., Pro. ruor of Mruchrmiury onl Nutritwn, ~)niversily of Puerto Rico ScAool of Medicine. Saw /uaw. JACK ('l1Al ON. M.D, A..o.wrt Prn/r.- s.r of Anrsthesioluq, New Yor\ Uni- versily Mcdreal Cenlcr, New York. CHII DREN'S HOSPITAL OF 1.OS AN- OE1 ES. Los Angeks. SANFORD C1IODOSH, M.D., Assiwnr Pro/eswr of Medicine. Tufts Univer- si(y School of Medkine, Ws(on. NAI lC•R 161. CHOPRA. Pe D., Pr.•/rs- sr a/ Che.niury, NarA Carolina AS- sicuhoral and Technical Su1q Uwiver• siry. Greensburo. PIIILIP COOPER, M D., Cl/nko/ Pro. /rssur of Swrery .nI Directr, Srrb .n! L.Aworory oJ Cellrlr Physiology Albeet Einslciw College of Medicine J Yeshiva U.iversily; Chief. Surtk.l Se.rirt, Veterans Adminiura(ion Hoa- pild-'1 Ae Brone, NY. PAUI. T. COSTA. /r. Pa.D., AuocMre Pro/rar of Psyrholofr. Universitr of Masaxtsvselts at Boslow, Doecbct(er. /O/1N /:. CRAICiHEAD, M D., rro%s- aor o/ raholo r, Uni.ersity of Var- nrrnl College of Medicine. Burliniflo.. ROBERT L. CRAIN. PwD.. Asslurw Pro/rssw ../ Suciolosy, UnivcrWy of CAicugo, Chicago. IRVING P. IRAWFOND. M D., Piofes- sor .nJ Chrlr„wn, Drpurrnrrnt of A//- ooA/elory. U.iversi(y of lowa College of Medicinc. Iowa City. SUIi BUCKINOIIAM. M.D.. Auisr.n( rro%srr of Pordiorrics, Cdumbia Uni- versNy College of Plyskians A Sue- aeons, New York. A. SONIA BUIST. M.D., Au.uWe Pro- Jecwr of Medicine .nf Physiology. University of Oregon Iledl\ Sciences CtMer, Purlland. BENJAMIN BURROWS. M.D.. Asso- c/arr Pro/nsor of Afrdaine. Uaiversay of Chicago. Ct.icaso. E. M. RUTr, M.D. Ch/r/ r.tAolotln. 1 oe Angeles County General Hoqi(al, l.os Anaek.. RICHARD ll• RYERRUM. PMD., ho- tssr of Chrndsrry, Mkhira. S1ue niversily. Eas1 Lausi.r. SISTER M. EMILY CAIIIL/, Pw D, CAornr.n- Dr~..trnent of Chrmisny, Reli. (-ulk{e. Weslon, MA. BRUCE P. CAMERON. M O., PN.D, Howard Ila~bes Ins(i(We, Universit~ of Miami Scbool of Medicine. Mi.rnl, F L. EI.ROY T. CAN7 RELI., Pe D. Chair- m.n, Dr~.rrn.r.u of rh.rnwolorr. Tesas College of Ouropathk Medicine. Nwr\ ltsas Srae University. Iknlun. 106 T. TIMOIHY CR(X'KER, M D.. rro%s- W11 1 1AM O. CLARK, Ps( D., Di.e.Lwt s,r of Meduiwe. Universily of C.N- r.y.h..Phrnwculorl Rrserch Lub..r• fania College of Medkine. Irvine. tory, Veterans Administration Il.npilal, Scpul.eJa, ('A. CAKROII. Y. CRU1S- M.D., Aruniurr, riufrs+..r ..1 AlrJiunr rnJ llumrn HANS T. CLARKF., OSC., rr.•/r.ur o/• Phrsinl..Kr: /hrru.r- Sr.ri..n .. Pul- Bi.rhrrniury, ('ulurul.ia 1/nivcs•ny powwry AlrJniae, Univer.ily Cali• C.ulkrt uf PAysicians i SurAeun+, Ncw /urnia ScMwA of Medicine, 1)avis. York. CECII. E. CROSS. Rese.rch Dr~..rnwwt, JAY U. ('OFPMAN, M D, Se.tinn St. /uxpA IlusPi(al, Burhanf,,('A. IleoJ, rrnPhrrol 1'rs" rJw oeFrtmrwt, UniversNy Iluspilal. Bu.ton. DAVID W. CRUMPA('KI K, Pu.D, ho• /nsr .nJ Chwirnwn, Drp.rtnrrn( ol AI I I:N 8 CU1/1:N. K1 D.. IhaD. A.- Enrkonmrntd, ropulal..n .wd orp aado, ~kChir/, ~k 1)ioloAy, Unrversity of Cd NN Wte rru/nsr of AIeJ.. ine, ~ Pulnrw.rt Srdlu., Tewrpk Umversily bo•rWer• Sciences ( enter. PliIladclphia. AI.BI`K f DAMON. M t)., Pa 1) , L rc• DANIPI COHEN. D.V,M., M P N,~( .• trrrr on .Inrhruruloty; Rru.rch Asso- arrrant rrn/rssw of Vrtrnn.ry fPi• rwrfr ln AfrJlcd AnrAr..pdorr, Pea• cknrnAur~ onJ Public Hra1rA- Umver- ~y Muserm- Ilanard Unr.atily. sUy of Pennaylvaaia Scb.wl of Veler• ( a"`I°iJrc• A1A. inary MeJicine, PbiladclpAia. ALLAN C. C(N.LINS. Pw D., Asr.xi.te rro/rssr of rhrnacoln y- /nsliudc /ot 7 HI)MAS R. DAWBF.R, M D., Assorlate l9u/risw of A1rJi.rnr, Bouon Uwiver- sily Sdsuul of Medicine, B.„lon. Belavioral Oenelics- ~)niversily of K F. 1)AWYON, PN D, Profr»or of Ior- Colorado. Boulder. nnt. Columbia Universiry- New Yoet. 107

I JOHN P DELANEY. MD, PuD, As- s..rwrr Pro/rnr al Swge.y. University of M.nnesou• Minneapolis ANDRI-W S DIBNER, PN D., Earc- rn.f, Psrcho Rr.rerch. The Age Cen- ter of New Fnlland, Inc, Rouon. f:DWARD F. DOMINO. M D., Pro/n- s.w of PAwne.ruf..fy, University of MKMgan, Ann Arhor. RAI PII L. DORFAIAN, PN.D, Directer of Laf•.Netr.rr, Wotcesuer Foundalioo (ur F.PeritneMal Biology. Sbrewsbwy, MA. H FRID 1)(1WNI-Y PMI), Aniscent Pr../f.N•r .-j Phyrwfofy- University of lc.a+ Ilcahb SCKnce CCMtr at Dallas; I11If.NM, (WJWI.INHfYr RfNW.A,Car- d.uPulnNamary Imlwuwe, McthuJi+a Ilo.- Pda1 ut 1)allss• Dillas JAMES I DYAR, PwD, Asduwnt Pro- /ruw of Rwlotr. Bellarmine Culkge, I uuisvdk, KY. RI('HARD H. PARI E M D. Chkl. Pufnro..e.y F'rncnon leluret.ry: As- s.stenr Pro/ensor of Mrdirinr. Uai.et- sily of ( Aicego• Cbuaao. JOHN W. FCKSIE/N MD. Auutewt Pro/rslor of Inu.nnj* Alrftr.wr, Suu l/.iveruty of luwa College of Mcd.- ane, lowe Cuy. BERTRAM F/C111 1., D D S.. D.rfcrw, InuirWt of Ston.atul.Kirel Rraer. h, Science Resources FounJalion, Waler- lown• MA. HYMAN ENGEI BERG. M.D. Aftend- iwg PhyNrfon, C'edars of Lebanon Hoe- Pilal, I us Angeles. CARI.TON K. FRI('KS11N, Pu D., Pru- jr.N.r of PhrrnNn.d..ty. The University u1.Tetas College of Pharmacy, Austin. V(il NY. FRWIN, Pu D, PralrtYw of Phw.ntw.d.r~y: Dren. Universily .d ('ukrado SchwA uf PAarmxy. BouWer. HFNRY 1. ESBFR, PN D-, Research Im- nrrnoforirr, Mason Research Inuihu/e, Wur.e+ter, MA. WA1.11•I( B kSSMAN, MD. PMD.. Pr.•/r.Nw .•/ Lnh.d.qy rnJ Rlu.Afnt• utrr. 1)utrns /•dlr/e uf the (lly Ilni ve.uly .d Nr.• 1 N.k, I ln.hong JOHN R. FSIERI.Y, M D. Associate Pro/rrs.w of Prtholory. University of C'bicago Pri1t\er School of Medieine, Cll/cego. HUGH E. EVANS. M D, Daraw, f)r- prtment ..j PrJN.rrks• Jewish H.nPi- ul and Mcdical Center of Broollyn, Broullyn, NY. HANS l. EYSI~NCK, PND- DSc., Pro- /rswr of P.yshofogy. Insliwre of Pay. ehiatry, University of I unJon, Loo- doo. HANS 1. FAI K, Pe.D., Adjunct A»ocl- Nr Projrssor oj PrAofoyy. Universily of SowAer• California School of Medicine. I oe Angeles. DANA 1. FARNSWORTH. M D. Nenry K. Uln-tr Projfssor of H/eient and Dirrctor, University //relrh Services. Harvard Universily, CansbriJlie, MA. GAD FFINSTEIN. PN.D., "rnior Lre- trrr in S/ochnniury, Tbe George S. Wiu Center of Life Scienccs, Tel Aviv Universily, Tel Aviv, Isr.el. FRANK C. FERGUSON. la., t.1.D, CAe/rnraw. Department of PArnurol- gy, TM Albany Medical College of nww Ilniverury, Albany, NY. THEODORE N. FINI EY. M D, Diree- tw, Pr/wwnwy Research te6ororory, MowM Zion HoapiNal, San Francisco. WILt JAM 1. FISHBEIN. /f O, CAief oI E~Jrntwloq, Chicego Board of IltahY Chicago. EDWIN R. FISHER, M.D, Director of LaAwaorits, SradysWe Il.xptal; Pre- ,nrar of Pethofotr. UuiversNy of Fi/1sbrAb School o( Medicine. Pilta burgh. RUSSELI. S. FISIIER, M.D, University of Maryland Sc`arl of M<dicine, Bal- linuwe. WILLIAM 11. FISIIMAN, PN.D., Prnl- JrM, La lolla Cancer Research FounJ.- liun, I a/ula, C A. B L. FRFFDI.ANDPR. M.D.. Directr of ('on.err Resrrch, Mouta 'L-wn Hoa- PiNd and Medical Center. San Fren- cifto. FRFt)ERIC A. FRENCH. A 11., DMrc- rr of Cen.rr CArn.othfrepy Rner.A M.Nint 7wn Ilosprr/ and MedicJ ('enler, San I-raneisco. I JACK FRI:UND, M D., A.tLton/ Pro- jrssor of Ph.rn.ecoto[y, Medieal Col- kge of Virginia, Richmond. LARS FRIBERO, M D., Pro/rswr ow1 CheMman. Drprnnfnt of Environ- nrrnt.l Hy~irnr, Tle Kardiaaka Irrui- WIe. Sloctblm. Gil BERT It. FRIFDELL, M D., CAk/ of PorAofo~', St. Vincent Hoslild, Worcesler, INA. GARY O. FRIF.DMAN, M.D., As.btent D4rcro., Dr~..rwrrnre/ Alrlicd MerA- ole ReurcA, Kairn Fowdallow Re- seard Institute. Oakland. ('A. I/. /1U011 FUOENBERO, M.D., Pre%s- sr of MtJkine. University of Cali- fornia Medical CeNer, Saw Frarcisco- Pro/essor of Iocrrrb/ory ond fnrwrr- nofoq. University of California, 1lerte- My. ARTHUR FURST, Pu.D, Dirertw, ln- stiteue of CArwtko/ faiolory. Uwi.eraily of San Francisco, Sata Fra.cisce, MURRAY B. GARDNER, M.D. Asso• clae Professor of Pathology. Uni.er- sily of Sowlser• California School aI Medkine, Los Angeles. GEORGE O. 08Y, M.D., Dlrector, rA.- nry-Howrll Cennrr RnrrrA I ot•ar.- bryt Associate Pro/tssos of Sr~~er~, The lob.s Hopkins University Serool of Medieine, Rahinaee. RONALD W. OILLETI'~ lsr1.D., Dkrc- Ior, lesk Science ReswcA Unb, Caw- oer Cen/et of Hawdl, University o/ Hawell a( Mano., Honolulu. GFRALD 1. OI.F.ICH, M.D., Grwsdtonr in Helicine, ReseaeA Lalwralory /ot Alkrsic Diseases, Mayo Clink .nd Foundauon; Pro/r»ar a/ Intrrnwl Nedkinr and ,ntnmwnobryrv, Mayo Medical ScMwl- RucAesler, MN. TIIOMAS M. OlK'KE, M D., Asso.-1ne PIo/rrMM of Prrventirr Mr/klnr wnI Cornmrnuy Health. SeWn Han Col- kfe of Medicine anJ Denusuy, /asey Cr/y, N/. DAVID M. 001 DF.NBFRO, St-D., M.D, Assw'wrr Pro/euw ../ Parhu/- ..Ey, TenPle Univtrshy IleatlM Sci- ences ('eMer• PAiladelPhia. PAUI (N H OI I ABI? R- D D S, A sNr uu hu/f.NM o/ Prn..J..wr..l.•Kr. IlarvarJ S.A.NA uf Denial A/eJ.c.ne• Buuun. LEUNIDE G(N.OSTEIN, D.Sc., Asso- ri.re Pr.•jerwr of Piychietry. Inuilwe fur Mental Hedr` Sciences, College of Medkiwe A Denlislry of New /ereey, RrlAers Medical School. Piscalaway. IRA GORE. M D-, Pro4asw of Pribf- ofy. (/ouon University School of Medi- erne; Chk/ of L.Borrory Service, Veterans Adrni.islralio. Ilosple4 WeM Rotlwry, MA. JOHN W. OORROD. D.C.C., Lrcrwrr /n IwPArm.c•y, Chelsea Colkge, Uni.er- .ily of I ondoq London. OFRTRUDE Y. OOTiSCHA1.L, Pu D. Asrbrenr Pio%ssor of /IocArndseq, Colnnsbia Urdversily College of Pnysl- cians A SwKons, New York. A. CLARK aRIFFIN, PM D., Ned, DeP,rr.wnr of 1lorhendsny, M. D. Anderson Hospital aod Turnor InN- Iwe. Uwi.ersily of Tetu Medical Cewler, Houston. ARTHUR L. GROSS, M.S., Senler tf. rArnNU, SowMwesl Re.euc\ /nrMM., San Anlowioq TX. MORTON 1. GROSSMAN. M.D., Pm.D., Assoeiete Cliwkmal Professor of M.M- ew. University of Calirornia Medka) Cew/er, Los AwRda. CARL C. ORUHZfI', M.D., Ih1.D. A.• sotArti in rhy.IoloRy and PAriwn+l- oRy. Uni.ersilY o/ hnnsylvanla Ore& wlo Sclwoi of Medkine, Mi1de1llW. JOSEPH 1. GUARNERI, M.D. Arqnd- iwP MkrotNolodier. DMener. M/eroM- P Ldor.rarks, tong Island kw- HA- bide Medical Cewler, Queens Hospital Cen/er A/RN.1bn, lawaica. IN1OA A. Ol11RO1S. Pw D_ Associate Prn/esvn of !'..nrMrwniry owd EwrMen- .nrntel Alydw int- University of C.II- fornia College o/ Medicine. Irvine. FRANK F. OU7HR/P., PaD. Pro/rs• sw of F:ntonrolo~y, NorrA Carolina Slale Colkge, Rakid- 11. B. HAAG, M.D., Pro/rssor o PAr- nar.doty, Medical CoBqe ef ~lkgiwl., Richmond. F. /. I/ADDY, M.D. PM D, Pre%sar enJ CArwnr.n. Dep.fr«trnf e/ PArsi- .d..ey. Ilniveruly o/ Ot<IJonu Medical ('enler, Ollarwna ('ily.

IOSNPH N. HAFKFNSCHII:L. M D. Drrrcro.. crdioournrowarr frnir, The L.nkenau IlosPiul• Aa,oc/orr /w Medi- ciwr, Ilniversiif of Pennsylvania School of Medicine. Prila.klOiia. RIAkO1T HAMOSH. Pw D.. Rr,rrrA Associ.rr, Drprrmrwr of rhlsiolKf ow/ IiooAysirs, Oeoroeto.n Univer- ailr. SAools of Medicine and DeMia- Irf, WasAindro., D C. BERNARD HANES; PwD., rro/rssr of Hr.IcA Scirwce, C. iforwi. SsaN U.i.ea- ritr. NonMidp. RICHARD 1. /IAVFL M D., Au/srw rro(rs,or of M.Jrrrwr, UnhersN' .f Cd,fornia Medical Ceaer, San Pru- CIKo. IIERSFRT R. HAWTHORNE. M D. CA.nwun. Dr~..nnrwt of Swvrry. Uecversily of PcnMyl.awla OraduaMe School of Medicine, Pfiladelpia. /OIIN A HAYFS. M D, Auocl.rr LrAulot.sr, MNlory InstaWe of Pa- rholotr. Sowo. Cny Hospual, sosro.. CLARK W. 11@ATH, M D., Pro/ruor of Mrdw iwe ewd Drrrr,or of Nr.hA Srr.- Icrs, Tulls llw.venny. Medford. MA. PAULINE HEIZNR, Pw D. RrsercA Associ.ue Iw Crrolu y and C)rorAewt,- rry, San Francisco ~euiwte of Medical Scknces. Saw Francisco. CAROL 1. HENRY. Ptr.D.. Dirrcror, De-~..nmwr o/ E.Prr4wewwl Owcolo~P, )r/krobiolotkd Assoelueti IsK, M- Iheda, MD. . IIERSFRI' ff IIFRSCOWITZ. Pw.D., A,wrlut /ru/r,Nr of Mkru6iwdcrty. Gewgeluwn U.iversiwy Schools of Med• kine and Dcalisuy, WasAinpon. D.C. LAWRENCE L. HESTER, la, M.D. Professor owd CAdrwsow. Drprln,ent of Olsrrrrirs and Cywrcolotr. Medical Colkp of South Cuolina, CMrks/on. BoRE CIlRT1S HOFF, M.D., hM D, rro/ruw .wd CA.irsn.w, D/v/sAsw of PrycAierrk Research. Medical College of V,rtinia. Rkhmond. RUSSELL L. IIOLMAN, M.D., Loulai- .n. Srare UniversMy School of Medi- eine, New Ork.ns. 01 F A 1101 iFRMANN, M D. Re- rr...A lir.rwrru I..t.rnJ f.P.urnny, I ln,.r..,rr ..t N.w.. U.rne Nuue It.r..a IN FRtUDY NORItlUROI:k. M D_ t•rrsi- ATTAI.I All KAPPAS. M D, lrn/r„or CHARLES W. I..BEI.LQ, PN D., Assist- Jerv rnJ Dnr. f.w, eiu kesearch (mli- ewd Srnior I•ArNcr.w. The Rockefeller owr Iro/rs,w of Enrirowmrwrwl Hy- luwe. Inc.. C'amhridte, MA. UniversGy, New York. ~nt , lellerson Medical College. PWa- RONF R f w. HULL, Pw D, rru/ruor IIkATCI/ KASPAk1AN, M.D., Assi,r- ~ Ju,ulutNd Scirncea. FlwiJa Slate a .nr Dar.rw, CarJiuer,rrtw LaAwo. AARON 1. LADMAN. PMD., Pro/rs.or .er sil y. 1 aRaaa saec. ni ro.y: )n,.rw ru. in Mrdiw inr, H.Ane- .wI CAoirmaw, Drp.rrmrn/ of Aw.r- msen hfedicd College and Hospilal. ony. The UniversiNY of New Meaioo RONALD R. HUTCHINSON. PN.D., Rr- PhilaJelphia. School of Medieine, AlhsrRuerRue. ae.rcA DYrcror, Foundalion 1w fkYa.- Iwd Research. Autwla, MI. EI.IHU KATZ, Prr D., Auo.lnr trro/rs- THOMAS C. LAIPPLY, M.D. ho er .w of Sociology. Unlversilf of Chi. ,ur of ir.rA NortKrvessir. Sad- IIT RESEARCH INSTITUfF_ Clkato. cato, Chicago. versiry Medicd~Rool, Chicago. ALPHONSE 1. INOENITO. Pu D., Asso- SIURLLrY L. KAUFFMAN. M D., ho- MICIIAFI. E LAMM M D t•ro/e,loro/ rr.tr Iro/r„or o/ rA.rwr.. ulogr. Fast lessor of rarbolot7. Sute Univaaily . . ., t•rwAol.Kr New York Uwiversil Medi- Crdlne UniveraM' School of Medi- of New Y.wk Iw.n>,are Medical Cew- . y cal Cenler New York eine. Oranvdk. NC. Ier, er.wklyw. , . $6s D. HARRY L IOACHIM Attending PAUI. S. I ARSON PN.D., H.. ho%s- . , , eno. II,R IlospNal: C/inl- rrAologi,r I ANCEI. KEYS, PN.D.. Dirrcwr, Labor.- sor of IArnroc.i~otP, Medical Colkp , . ro/ Professor o! /.rAolog1 Columbia rwy of lhyslolotkel )fytirnr. Uraiver- of Virginia. Rkbnosd. . Ua.eray Colkte of Plyskians A sitr of Minnesola School of Public Sureeons, New York. Ileallh, Minneapolis. ROGER K. LARSON. M.D., CA/r/ o/ Medkiwr, Fresaw Coww 11oyMQr GFOROE /ACOOSON, M D. Pro/ruor IOSEPH R. KIRSNER, M D., rro/rasor Fre+rw,CA. .wJ Head. Oqrrnwwr of R.diolott. i .,/ M.dkiwe, Uwiversi/y of Chicago U.iversilr of Sowrer• California School of Medicine. Chicago. GUSTAVE A. LAURENZI. M.D., CAtr School o/ Medicine. Loe Angeks. of Mrdreiwe, SI. Vince.e HimpluL EDWARD 1. KI.AItlF.R, M D., Srwior Wo.eesler, MA. 1F.RRY HART JACOBSON. M D. Dl- Scientist. The Wortesler Fuuswlaliun (or rrcrw, Drruiow of ElrcnoPAfslolotr. EaPerimenld Biology. Inc.. SYre.n- JOSEPH Ir:. LAl/WERYNS. M.D, New York Eye and Ear InArmary, busy. MA. PM.D. rro/rs.ur (hdinrun.wd CAdr- New York. .w, Drprfw,enr of rrAolo~P ewd 1FROAIE KI FINFRMAN, A/ D., rr,- AtkroaeoPle An.towq, Ea'erinreMal JULIUS 11 JACOBSON 11. M D.. Asso- /ra,.ar rwd CAwirwww. Drpsrrwerwe of L.botalory of P.Imonary HrMOprAol- ci.rr rro/ruw o/Swerry anJ Direcrw PrrAolatr. Muum Sinei School of Med- oRr. Knlbliele Univeraheil u Lwves. I Swtird RrwrcA, UnivcrsiaY of kine, New York. Lsr.eq Rddlrr. ~Sr~ MURRAY E. JARVIK. PN D., Asroclare rro/eswr of Pharmacology. A16ere Elr PETER /1. KNAPP. M.D.. ResrrcA ho/rs,o. of PsycA/errP. Roslota U.1- verrilP School of kledidae. sosto.. ~ IAMES C. I FF., Pw.D. Aas/aowr ho/.a +w +/ RNa ~miurP. ~ainl Louis School of kledicissit. SI. Lisoih. Meie College of Medicine of YeJir. KENNETII P. KNUDTSON, Ui.D. Ud- ED~VARD LEETE. PM.D„ D.Sc hda- Unhenl/y' The Rrona, NY. veraiaf of WashinNo. Sctwd ot Wedi- r. Univenlly o~ Mi.r- ~ site DAVID A. IOHNSON, PH D.. Aul,rowr eine. Seattle. . ~a _ ~ K lo~e Pro/essor of liocAemisnl, I'a+r Te.ees- RICHARD A LERNER AaocA,re M D see S1are Uni.ers8y Cdkfe of Med- ALVIN 1. KOSAK. Pr.D.. Associri . . . . Merwfrer Serippe Clinic a.d Rese.rcY k{.e, lohnsun City. ho/essor of CAnwinry, New York University. New York. . FowdeUon, L.loNe, CA. OSWALD R. IONE'S, M.D.. SL LuIeY CECII.B I.EUCHTENSEROER, ft.D, Hoyical. New York. RObFRT A. KU/IN, M.D., A,wri.tr Rrod. Drp.rrn.rwr of CpmcAewslrry, Professor. D,rbiuw of Nrwo,wtrrP, Swiss IeslilWe for EqarkneMi C.w, W11.1 IAM 1. IUSKO, Pn.D . Arsocl.re New Jersey Sute College of Medkine, eer Research. Lsu.awne S.IraerlaA•. rro/r,sw of lArmecrwrk,: Dirrcrw, lerser Cilr. . , Cl/nicol rArw,.coAiwerks LaMwoaorr, lAY A. II:VY, M.D., .lswr/.rr Pro- Millud Filinare /Ioyild, RuRdo. STIO KUI LANDER M D Professor /r»om of Mrdinwr: ResrrrA AssocL PM D KANDUISCH ANDREW A . , . . ond CAoirn..n. Drr.rrsnenr o/ O6urr- wre. Cancer Researclir Insrirue. Ud- , . .. . The lacksow Lalors- Sas ScienNU rks ewd Gynecolotl. Universily of versl/~ d California School of Medl- , ewy, Bar IlarEor, ME. I.u.d, I..nd. Sweden. tl~e, Se. Frawckco. PAlll. D. I F WIS Srnww I rrrwrr M 0 ARNOLD R. KAPI.AN. hrD., Dkec- MARVIN KUSC./1NER- MD, New , . iw NiuuparA..b.R., Royal Poutr.duaN Iw. l..s.•ru/w) of Mrdirel rewrrks, York Unlverury Medical Cenrer, New Med,eal SchaA, Hammersmah Hoyl- (-1e•elanJ PsYckvau,e Ir stUwu and York. lal, 1 orwl..n //..cput./ le•eland 111 Coikp o( Medicine. Rur-

AVERH.L A. LIESOW, M D, Cliair. rn.n. Drp..rnrra of 1atAoloty. Ysle Universuty School of Medicine. Ne. Havnti C1. IRVIN E. 1 IF.NFR. f N D., rr.-lraor o/ /torAr.niury. Ut+iversdy of Mmne.ot; Sr. Paul. ESTEN O. LINDSETH. M.D.. rw.D. 31. losqb's Ilospul Reseerch L.boe.lary, Sr. Pwl, MN. ROSERT H I.INNIa L, hM D Assoedf ut rrofr.wr of l'A.ndurl* doirereNy of Venwowl, Surlintro.. HERRERT L. I.OMRARD, M.D.. M F-H, Asd..rr, ('.wrrr RnrrrA h- mrru, New Entisnd DeKoneY //os- Nlel, Ib,ru.. 1 P. LONG. hr 0. rrofrssar of rAr- snoro/utr. Sure Uwiversir' of k~.. Colkte of Medicine. lo.e Ciry. CLAYTON O. 1 OOSI 1. rw D, IS( D, Nurb.ts rrofruur of Mrd.rinr ..d Ltboloty. Uwiversirr of Sout`ertt Celi- /orni. School of Mediciwe. I os Aqcks. DONALD 9. LOURIA,,M.D, Assoriw rrofrrsor of Mrdurnr. Cormell Ura.et- siry Medical Colkp, Nt. York. KENNETII MERRIIl LYNCH. M D. Sc.D., LL.D, rrofrsso. E.wrrnrsv o/ r.rAo/oty and CA.nerflo., Medical Cdkte of SowR C.rdiwk C4rksro.. INES MANDI, ItM D, Assistont rro%s- sor liockrmistrr. Cokwwbit, Uwieetr sAy pd nysiciur & Swtews, Ne. York. JOHN H. MANIIOLD. Is.. D.M.D rraten~or osd DYertor Drprrnwnt ej rotAo/ugy ond Orol biotnosit, Ne. Jersey Colkte of Medicine .nd Dea- lislry, Ier/ey Cily. DAVID E. MANN. hr O, AuocWe rro/ruar o/ rflrnuscofoq, Tewde ' U.i.asiwy School of r1.nn.cy. H1.- delphie. FRANK ARTHUR MANNINO, M.D Anisr.nr Professor of Olstmks on~ cynrndofy. Waoew s HoapM.l. Los Angleks CorsrylUniversAy of Sow`eew Cebfarw. Medical Cewrer, Los Aeteks. JOHN P. MANOS. M D. fnsrrrnr in Ywulury r+d e.rrr.iuloty. Medical ('ollete of Soutl ('arulins. Chsrkuon. CIIRISTOPIIER M. MARTIN. M D Asusrwnr 1'.u/rss.w of MrdNnsr .nl Dnnt.w. Dir.dun of Infrarwrr Dis- r.xs, Scnow 11.11 College of Medicine. lersey Ciry. R. RUSSELL MARTIN, M D., rrv/etsor of Medicinr, and MirrokoL.r~ l Ina nrwoloty. Reylor College of Mrdicinc, HouMow. REGINALD O. MASON. M D.. hM.D. nsor and C/rarnwin, De~.r rnvnr N /.wAulo ot~. Univeesirr o/ Uu~ ( d- /er of Med.cine, Sdr L.ke Ci.y. MASON RESEARCH INS7ITUTE, Worcesrer, MA. DONAI D/. MASSARO. M D. Auorl. .rr rr../rssor of M.d.rlne. Oeo.r weskrtylow Urri.ersiwr Sclwol of Medicine. W.sAing1 DCo.. CHARLES McARTHUR. ru.D. rsy- ckolotir, Un/rnsiry Neobb Srrrkrr, Henud Uwiversity, C.e.pridte, MA. CHARLES S. McCANTS. Frr.D. Asso- tWr rro/essor of SoQs, Nonh Cttts li.. St.M Colkte School of Atricul. lwe. RekitA. GERALD E. McCLEARN M D., Di- recror. Inuirrre for Rrllioriord Ce. sYrKe, Drrr rnwnt o/ r+yeAoloty. Uni- eeesily of Colorado School of Har- w.cy. Rorlder. HENRY C. McGILL. )tr.. M.D, ArtinF Nrod. Dry.wrnwnt of rrtlwlucy. Lour wwe Slwe U.iveesiry ScAoo' of Medi- cine, Ne. Orleans. HENRY D. MeINTOSH. M.D.. rrofes- sor of MedkiMe ard Director C.rdo- ...crlor L.1ar«o.y Drb 6nivenity Medical Ceaer, Drrrk.re, NC. FORDE A. McIVER. M D., Associate ho/rssor of Pathology. Medical Cd- kp of Soa! C.roBsw Cli.rksloa. EDWARD McKEE. M.D., rrofrssor ood Acting Chairman Drpranunr of Pathology. Medical C=olkte of South C.roli.rt. Claerksuow. KELLY T. McKEE, M D.. Associate rrofrs.or of Medklne, Medical Cd- kte of South Cerdiw.. Cbarkaow. HERBF.RT McKENNIS, JR.. M.D.. Rr- wr.A rrufrLUr of Pathology. lJniver. sirr of b/ieari School of Medicine. Murni. F1.. 112 VICTOR A. McKUSICK, M D., rrofis. sor o/ Mrdicine. The 1okawe Ho~kira U.i.asiq SJool of Medki.e, .a8i- rwore. ROSS L. McLEAN. M D.. Associate rro/rssor of Medicine. F.seory Usi.er- sMy School of Medici.e, A11.a.. WILLIAM F. McNARY. 1.. hr.D., As- sociorr Professor of Aw.ronry. Roslota U.i.ersily School of Mdicine, RoMOr. NEAL L. McNIVeN, ft.0. Tw Wee. eesrer Forrwd.liota !or esterirtaeM.l Riolotr.SAe.nlury. MA. HANS MFIER, D.V.M.. Senior St./ Sri- rnNst, TM lecksow L,bwaory, Rer I/erboe, ME. JULIA MEYER. hM D.. Assocl.rr rro- (rssor o/ Oral r.tAoloty. Uwersky of IRiwois Colkp of Den_jrry. Ckk.p. DOV MICHAELI, rND. Assistonr hs rssor of l/orArndstry ond Sr~..y wiversily af Califor.i. Sc~od o~ Medici.e, Sow Fr..cieee. MiCROBIOLOOICAL ASSOCIATES. INC» Bethesda. MD. BERNARD J. MILLlR, M.D. Auissw rro/essoe of Awwonry, le/er.on Med1- eal Celiqe, Mil•Ae1Ni.. JAMES O. MILLER. MD, tr.0., ho- fr»w o/ rqckietr ood rsyrhe/e~~ Director Mentol IJroltA RrsrrrA )w- stitsre. bni.eraMy of Mkw.M, An. Areor. lERA1.D A. MITCIIF.t L, M.D, Aswr clar rrofrssor o/ Anaorwy, W.y.e Suk UnioeraNy ScA.rol of Medicirrc, Detroit. CHARLES MITTMAN, M.D., Dirrnor. Dqortrernt of Reqir.tury Diseases. City of 11o~e National Medical Cew- ler, Ddtarle. CA. H1/(ill MONTGOMERY. /d D., Asso• ri.a rr.dr.sr of Mrdkinr UeiversAy of Penn. l.ewie School oi Medicine. Philadelphia. • Is. I)'N. k1ONTOOMERY. le.. M.D.. rrn/nww of rahofotr, llniversiNy of Te..s SowA.e+tern Medical School. D.Iles. GEORGE E. MOORE. M D, hr D. Di- re. hw, Ros.ell hrt 111canori.l Imri- Iwe, M.Jleb. NY. i KENNETH M. MOSER. M.D.. Ass4wnr rro%ssor of Medkine. Oeorgelo.. U.ivenily Medical Sckool. Wshiy- low, DC. HURLEY LEE MOTLEY. M.D., rrefes. so+ of Mrdiriae and Director. Crd/o. Sow ~lra~w C. if~iri.r School ~ of *M cire, Los Aqeka. EDMOND ANTHONY MURPHY M.D., Sc.D, Aesoc/.a ho/essw .i Ibsraiulcs owf l/r/kiv. T1. loR.e cHiw, ~e Relliinote University School of MoY- WILLIAM 3. MURRAY. Sc.D. S.w/r Staf Scientist. The l.cksoo L.kere. lory. u.r H.rbo., ME. RICHARD L. NAEYIP, MD., rrofessar and CAdrwr&% DrNrrnune of rrAol- oty. Fesuylvui. Slete UoivesiNy Cd- kte of Medicine, Hersmy. OEORO R. NEURATH. hs D., 11lk... on.fytic.l trbrotay, Harnbwt, West Ck-y. ALBERT H. NIDEN. M D., rrofeer../ fifrlk/nr. Draw roswp.Aw1e Medical 3AooI .ad Usfiverswy of SorRer. Cdi/er.i.: CAie,. r.rn.ew.ry Db..a Srctioo, M.rlis Lwkmr Kiq IlesEil.L Los Ayeks. OAK RIDGE NATHNNAL LAVORA. TORY, O.k Ridee, TN. DONALD M. rACE, hr.D., rroftn.r of rAysinluty and Dsrreror, lsritw e. Ciprdor ResrorcA, UwlveseNy •d a. Li.col.. BEVERLY PAIGEN. rw.D.. Concer Rr- sroreA Sa'Irntiu i'. RosweR PYk Me- waid Lairwe. Diagr.b. KF.NNF.TH rAIOEN, M D., Dirrnor. Drrsrtrrnt a/ Al.dr.wGr I/uloty. HenhR Rese.rA, Inc., Ros.reR rerk Division, Sr/la1o. ALRERT R. PALMER. tr.D.. Asduw rro/rswr of rwrholotl. Universily of Takdu. l ukdu. OII. ROSE k/ARIF. rANORORN, MS. As- siurnt F...d Tr.hwolot/st and lrrtrn. Dr~.rtnwnr of Food Science and lrrA- nulut_e, Uwiversily ot C.liforni., Davis. IOHN W. PARKER. M D. Associre rrofrss.w .r/ rrlAol.rty. U.iversi/y of 5.rwhee" ('.lifornu School of Meali- cine. ( o. Awteks. 117

1 I y..~ ~ #-A 1 1 NJ MARY STEARNS PARSHI FY. t•N.D.. Aniurnr Professor of An.wnry in O& rtnrkt .nd Grnrrulotrt ColYnlbia Univertity Cdk~e of Phyocians fa Sw- gewe. New Yrl. EDWARD W. 1 FLIKAN, M D, Cbir- ne.n, Drpru.rwr o/ f hrn..roroq enf Eipnnren/d TAtryerrkr, t<oelon Univerdry School of Medicine, Iloe(oa CARI. W. PIERCE. M O, fM.D.. f ru/ir w.r ef r.rhuf.K, wnd MkruAlulur)-fav- nrrndotr. Wnlwn~1on Universilr Sclool of Medicine; Dbrrnr of rrA- •foff and Lbrorury Mrlk/nr The lewish Hospital of St. Lo.ls, St. 1.oeais. MAI COI M C. PIKE. Iw D. Professor of Co..wnrwrtr M.Iklav and redwt- rkr. University of Sorlher. Califorwia School of Medicine. Los Aqeks. OTAKAR /. POLLAK. M.D.. Pst D. Eerc.rire Drrrtor, Dorer Medical Ro searcA (-emer, Inc.. Dover, DF. MORRIS 1•OLLARD, 1'MD., DMrrtr, LoNrn/ L.borNory, U.iversity of Nuwre Dame. Norre Daine, IN. C. M. POMERAT. PuD. Dirertor of Ablotkef Rr.erch. Pasadena Foasda• lion for Medical Research. (.sade.a, CA. S. N. rRADHAN, M.O. M D.. Iro/er- wr o, rhrra.rototy. Howard U.iver- sNy Cdkp of Medicine. Washi.Rto., D.C. H. R. fRATT•THOMAS. M.D. /ro/tr- aor of f Nho41 and Dtw. Medical Colkp of 3010411 Caroli... Charksbw. PROCESS i INSTRUMENT CORt•O- RATION, Broolly.. NY. MARTIN S. rROTZEL D.O S. Chief. DyortnvM of Oral rNholoSy, New.rk ( iNy Ile.pld, Newarl, NI. WAI.TER RP.DISCH, M D. Associate lrufruur of C//wirof Slydir4a. New Yurt Utdversiry School of Medicine. and NYU ResearcA Service. Goldwater Mee+orial Horpird, New York. TIMOTHY 1. REOAN, MD, Pro/rruw ot Mrli. int; Darrtor. Dirhiun of CWlWrapblr Dureae. CdlrSe of MrJ...ne end Ikno..rly of Ne. /erecr. Ner les.rr Medu.1 ScMuol, Ne.a.l WI1.1 IAM RF(iF1.SON, M D. rro/nror, oand Ch.irmrn. Dep.rtmenr of MrdKar Untulugl, MeJIcal Culkge of Virginia. R icMwnd. LYNNH M. REID. M D., Wolt.rh f'.o- /tuor o/'athofugf, I/uvard Mcdicd School. Souue; Chairrn.n, Department of lrholugy. CUdree i Ilospiul Medical Cemer. Rouo.. HOBART A. REIMANN, M.O. Pro(n- s.r er/ !<ftdkine, HaManawn Medical Colkge e.d I/osEiral, Philadelphia. ROLLAND C. REYNOLDS. M D., Ar- a/MW Professor of rMhorog). Univer- si1~ of Teaas SowAweuer• Medieel Sc~ord, Da11as. VICTOR RICHARDS. M D.. Chlr/ of Srrery, f•resbyleria. Medical Cenler, San Fraacisoo. ARNIS RICHTERS, Ihr.O., Associate ho/ruor .r/ f rholoer, U.iversily of Sowfrer. California Selsool of Medici.e, Los Angeks. WILLIS H. RIESEN. hr D.. Senlor llo- chrndu, ti/e SNewnr Dirb/on- IIT Re,earch In.tiune, Cfaealo. DANIFI. R. RIFKIN, fhr D., Assistant Prufru.w ../ Chrmkr/ Biuio`y, Tb Roelelelkr U.ivenqy, New Yort. R. H. RIODON, M.D. Professor of f.- tho4r~~~ Uwiversilf of Tetas Medical Rranc~- Oalveslon. SYDNEY C. RITTI?NIIERO, M.D., P.u tswr ol Lrteriebp. Uwiversi(y .I herw California. Los Ayeln. RENSON R. ROE. M.D., Associate ho- /easor of Srgtry; CMe/. Cardiac Sr.• g..y. U.iversity of C.Werni. Sclgool of Mediei.e, Saw Francisco. IOSE!t•H H. ROOERS, M.D., Holy Nswe of lews Ilinpilal, Oadwkn. AI.. ROBL'RT C. ROSAN. M D., Associate Professor of fotholody and reJiatrki, Sr. Lork Universily ScYool of Medl- cl.e; Airociett f'.thorog/u, Cardinal Okarae Memorial Hospital for C41- dren, SI. Louis. CIIARLES L. ROSB, Ihr.D.. Cl/nk Dbrr- tor; DMrctor, Normative Aging Study. Veleraes Adinimsiraflon (IwPatieM Chnic, Bouow. 114 I JOHN A. ROSECRANS, (•n.D.. Au..- rirr Professor u/ Ih.rnurul.Kl. Medi- cal Colkge of V"aSini., Virginia Cowo- nwwweail\ University. Richmond. JOHN R. ROWLANDS. hr D.. St./ Scknritt, Souwhwesl Research I.pirwe, Jan Anmonio, TX. BENJAMIN A. RUSIN. hr.D., Aasirt.nt Professor of rrilk Hrdttt, Baylor University Cdlep of Medicine. Ilor- sros. RONAI.D P. RUflIN. Ihr.O., Professor +/ r~~~tl. Medical CdkSe of Vlrsinia, Ricf.mond. HENRY 1. RIISSEK, M D rrtddrnt, 7)~e Rr,..ek Fo.ndaUots, i.c., Sratew IJand, NY. W. C. RUSSELI., M.D. University of Te.as Medical Center. Houston. WAYNE I.. RYAN. IM.D., Professor of /iochrraiury. University of Nebraska College uf Medieine. Orw.f... PETER F. SALISRURY, M.D. h(.D. Head. lnteNrire Trternrtnt Cenrer, Sainl Joseph Hospital. Burbank. CA. PAUL SAI.TMAN, Ihr D, Auiuwt ho- /tstor of Bbchendrny ewd NwNtlon. U.iversilr of SoeMrerw CdifarM. Schod d Medicine. Los Aqeles. ULRICH 11. tiC/1AE1•rl. M.D. Dirtc- ror of Ntwr.rRhrwurolop, Maww Ra search InsrNwe, Worceder, MA. IOR(;I:N U. SCHLEOEt, M.D. lM.D. rro/tuur and Chdrnasn. DrNrtnrewt 01 Sr rry, Td..e U.iversilf Sdod MAURICE S. SEOAL. M D., Crlwkd Professor of RlydkMr. Trfu Uaiver- sity Sebol of Medicine: Dirtctor, De- portnwnr of fnhor.tlow Therapy. B•s- 1ot, City Hospital. Sos1w. CARL C. SELT'LER, he.D., Ho.orry Research Associate. Peabody M.nws>. Harvard University. Cambridge. MA. LUCIO SEVERI. M.D. Director "/ Dean. Iwultrte o/ Awaewsr and fattal- Division e+f Cnrcer Reaeatch U.ivcnl(y .1 htsrRl.. M.RiR Italy. CHARLES R. SHAW, MD. Ch/tl. Stc- tion of Afrdkd Qenttks. M. D. AM- dasow HorEitY and TMwnor Iws(ie.N; rro/essor of aofegy. The University e/ Teas M Horrwoq Ilouao.. CHARI.hS E. SHERWOOD. M D. Ar- siaant Professor of Rodbfotl. Uttiwr- siy a/ Rochester School of MedieM. and Dtwriauy, Roefteser, NY. SIIOI/ SIIIRATA, M.D., h1.D., f r%} t.n rrwiworofody. University of Hawa~ii School of Medicise, Honolulu. DAVID L. SIMON. M.D.. fwanrrer ie fntrrairl NedlcMe, Cincirwll General Hospilal. CincinnaU. ERH: SKINHD), M.D., Chk/, DtEr/- wwar o/ Nerofup. Rl.yebjeq Ila*F 1.1. Copenhagen. NATHAN H. SLOANE, IhLD ho/t.• aer o/ /iorhewristrl, Ttte Ljdvenh~ of Teweeuee Ceoler for sh. Heahtt Seienees, Memphis. TIII?ODORE A. SLOTKIN hr.D.. At du.M Professor N rkarowcolKT. Duke h.wy NC rsilr Medieal CaMer, Drr• of Me4locine. New (kkas. •' OEORUfl W. SMETTERS, M.D, Au.- ALVIN R. SCHMIDT. Dkteew of Counseling. Trfu University. Med- f.nJ. MA. IAKOR SCHMIDT. M.D.. IM.D., Aa- ruto.u Professor of Aiochrrel.tr)- Di- rlrion of Biological Sckn.rr. Sure University ot New York at S1onr f/root. S1ony Broot. ISAAC S('HOUR. D D S.- Mt D.. D Sc„ Dean. Universirr of 111iawMs t-ulkge of DeMi>try, Chicago. SCRIrI'S CI.INIC AND RFSFARC H FOUNDA 11l)N,1 a /olla• CA. c1w /n raA,r/,rg1, Na(hwesleer! U.1- ve.sieT t,ledicd School, Chicago. OP.NP. M. SMITH, hr.D., A.risunt f r. [rasw e./ lrTeholoRr, Harvad Medical ScAuol. Massecb.aHU Oenerd Hoql- lal, R.ruue. LU(YI I! SMITH. 1'w D.. lro%rar__ e(• /iw htndnry. Darusowtt M IkMe ScMw1. llawwer, NIl. LOUIS A. SOLOFF. M D.. /l.wrb r. Lery DuNn4wishtJ Srrrkt rro/eerorr rr.r/n..w of A/rdw inr Datrrr. Rt- trrrh 1 ipfl tdwunry, TemUd- vetsiNr Ileahh Sekncea Cenur a- Jclphia. 115

I O+ ~ r m m J A W A SHELD(1N C. SOf1/MFRS. M D. Dirfc- tr of tibo..twifs. I enoa HiR Hos- Pild: Clurk.l rro/rnw of P.rAdop. Cdwwbi. University ('olkge o( t'Arsi• ei..s A Surgeows. New York. ERNEST S()NDIIFIMF.R. hM D.. Ano- riur Iru/rrwr of SrorArn.iuny. Cd- kAe o/ Foecslrr. Stne U.i.ersqT o/ New Yor\. Syr.crse. T. M. SONNFIIORN. PM D.. D/ulw. twsAr/ Srrrrcf Iro/f.sr.e/ Zoologr. Ldi..a U.i.ersily. Rloo.i.No.. SAM SOROF. ArD. Nr.l. Dr/+rrwvw( ./ Ahr.owsuJrr.l.r CAriwi'urf. T1» Lsri1.N (er C.ncer Reseach. MR.- ile1rY.. SOUTHWP.ST RESEARCH INSTI- TlslF.. San Aww.iq 1x. DAVID M SPAIN. MD. DMtreor, Do. p.rrnrrwr o/ l.rAol.rry. The lrueld.M 11.r.lw.l (-enrer. b.oodly.- NY. AI FXANOPR SPOCK. M D. Arduw rrolrs.r of rr/rorrkr. Oa.e Uwiver- siry Medical ('erwer. DwA.wr. NC. FRFOFR/CK I. STARF. M D ho/fa ar o/ NMrifio.. H.rv.rd University Sclwul u( ibWK He,hA. Owo. C. HAR(H D STPFFFP, M D. Di.rrrr of I,rMr.rwkr. MetAodist Hoyiul. Memo1J.. JACK ta. STRONO. M D. Arwriwf Pr..1rsYM of l.rAdoar. Lo.isi... SuNe t1.i.enNr School o( Medrci.e. "Ne. O.kaes. MARION 8 SULitrP_ROER. M D., rro- /rsuw .e/ CA.irw..n. Deprawwt u/ Deroraolop and SrpARolo y. New YwA U.iversNl-BeRevw Medical Ce.- kr. New Yoel. RENATO TAOIURI. Pat.D_ Asx..Wr ho/rasor of lsrcAoto~~. Or.du.1. School of wsMre.s Ad.dniNr.Uoo. I/urud U.i.ersiry. RoMo.. DAVID W. TAt.MAaP. M D Di.fcror. wdf,-IV.rtwr Lrwt /wnbwr.ll.iversMT of Co/or.do Medical Ce.ler. De..er. LYNN M. TAUSSIO. M.D.. A.>rrri.tf rro/ruor .nd Associue CA.irn..w. Dr/..tnvnt o/ t'riwrks. Arison. Ile./lh Scie.ce. CenMer. T.esu.. MARC D. TIIAA/P.S, M D. Srnn+ Rr- .rrrA fft(o.. M.Fo C1r.c Nd furr.. da.... Rushder. UN CAROI INE <tFDELL THOMAS. MD.. hulrr.w Enrf.ie.u of Afrdkinr. T6e IoA.s Hoeli.s University Sclad ot Medicine. l.hinwre. JEROME F. THOMAS. PM.D.. rro/fuor V S.wrrrr Ewginrfr/nf. Uoiver.i/y of dilarni.. Rerlekr. JAMES L! P. TOMAN. hr.D. rro/n- sw .wI CA.irn..n. DrP.rt.sfnt of rAr- wrc . Chicago Medic.l ScAool. t.- aIY.N or Medical Research. Chicago. ANDREW M. TOMETSKO. hM D hee- idrM owd DMrrNv of Rf,rrr A. l.ilro. Laborborks, Ud.. RocAesla. NV. IANET TRAVPLI.. MD. AtwcWr rro/rurr of Clink.l PArn.rolotr. CorwrR University Mdic.l (•o1kA.. New Yarl. I IE SHA TSAI. PN D.. Rr.rrrA Ae.. ei.re M r.rAo(ovr. Yale U.werury Scbd o( Medicine. New N.ve.. CT. OERAt.D M. TURINO. M D. Prolruar o/ ltlydirMv. CdwrAi. University Cd- kv of Mysici..s A Swoeo.q Ne. Yori. D. M. TURNER. Pu D., Head. Ay.rr- ww.t of DrrrR A(fuloNnw ..d Rl.- rAnwtrny. H.:1elo. I aDorNor e. E.- rqe. Lrd.. ILrrople. YorlJks. Ey- 1..d. UNION CARtrID! CORPORATION. N.cka Division. Oak Ridge. TN. UNIVERSITY OF SAN FRANCISCO. S.. Pr..cisco. UNIVERSITY OF SOUTHERN CALI- 1(1RNIA. t.os Aryeks. ELLIOT S. VF•SELI, M.D., ho%ssor.wl CAoires... Deprrwrenl of rArwrerot- •rr, hnu+ylv..i. Slare University Cd- k~e of Medici.e. MiNo. S. HenAey t~ledicd Cewte.. Herst.ef. BRANISLAV VID/C. DS.. Prolrrwr of Andonry. Oeor{ero.. University ScAouls of Medicine and Dcmiwrr. W.sN.yto.. D.C. ROMEO A. VIDONP. M D.. Associate /rolruw ../ InMAolo4r. Y.k Uni.er- .ilr ScAuo1 o/ Medicine. New H.ve.. PF.(tTFR K. VO(1/. hr D.. rrnlrnor of A(nr..l.i..1..Ky. Universily of W..Aing- Iuw 54" u/ Medreine. ScNde. IIA IRENE Y. WANG. PrrD., Assistant ho- /essor ./ ifosk and CNwkof Inrwrr.ol- soii1•Twd of MSo.1 C ~oli.~ e~C! kMadir. 8. D. WARNER. M.D. ho%ssor of Pd- rAdoR~. Sr.ae Uni.enilr of /ar. Cd- koe o1 MeAie:.e. Iowa Ci1y. SHIELDS WARREN. M.D.. DLrctr ./ L.Mrororirs. C.ncrr Rfsr.rcA LMi- rwr. New 4aRla.ti De.oo.es. HeaPl- w. .oero.. YASUSHI WATANAtrP. h(.D~ Aur d.rr Mrinier. The Wisl.r LMi~we ot A.No.rr ..d eidoh. t'fi./d/W. RAR<tARA K. WATSON. M.D. As.irt- .wt LrterldoelM. Msa.da.e(h Oe.- ad Hoyiul• Rrn..cA Auor/re In /.rteelofo~r and loawrwalogl. H.r- •r~ Me~ial SeUool. Soam.. L8E W. WATTENRlRO. MD. tro%r- a.r o/ IrAolory.~U.ireraYy of MM- .e.ol. Medic.l Sct.ook Mi..eqoW. JOHN S. WAUGH. PrD. hNraer CAr.s4arr. M.o.dwaM. Nattilawo Ted woloq. Cae.lrMp. RICHARD L WtxHSLER. M.D. CMw- (n.l PArsloloAla, Meada. NoaPNd Iwi1sM..1 Researct< PinsA.rd. JOHN V. rPE1L, M D. Asdrrwr hr /raae. of lfrdicl.r. UhirasNT d Calr eds Medical Csaa. De...a. A. WEINSTOCK. 1'rr.0, Rrsr.rcA 1i} cA.ndst. Life Sck.ces Dirirf.n. qT Rew.rcd IwNw.. CYeap. RUSSELL W. WELLER. M.D. PotAol- "lsr Mer.orW H .f Cbearr Co.My. We.1 Clscaa. A., A. STANLHY WBLTMAN. rrD. Aar eire rrolenor of PA.rwwcderr o.f RexorcA. RrooUyn CoRep e! P1r- wuer. lrooiIM TlY. SIMON H. WENDER. hr D.. RfseorcA Professor of flocAendurr. University of OAl.hawa. Norrn... DUANE O. WENZEL. hr.D.. rro/rs.er and CAoirnu.. DeP.rarenr o/ lArww c r o.d To.icologr. Tl. University of Know Scbd d PR.nw.cr. L.w- rsaroe. THOMAS C. WESTFALL, Pw.D. h.- rArr..co(o~ ~ Irssor •l r. Usim ~ d VirRi.i. Scbol of I~/edki.e, tler- Im1e..iRe. FREDERICK E. WHISKIN. M.D. C.M. Director. Di./dew o/ Be.MA o.I rfr- ,enditr Eqri/i1r(rw. Tb Atp G.be of New FngLN, Lc. OooWa. lAMBS A. W1LL, D.V.M. th(.D. ho• lesa.r an1 CWnwM. Drrvnrwr of veScre.ne. U.i.«rily d Wr ROOER 1. WILUAMS. M.D.h./ rsr. of CAe.r/atr); Drec(n.. Clqaw 1~«.. /w/o. /lecArwk.f /wrMrrf. TM Uaf{• ' vasNy o( Tes.r. Awri.. DANIEL H. WISEMAN. M.D. Asdr- o.r h essor./ rtfl.rrka. University al Southern Celifor.i.- Clsil4a.'a Di- + 1. ..itior~ .. Ayelee 6o..tr oe.eral - Hoyiub Los A.yelas. GEORGE WOLF D. twa., he/rsr o/ rAraW.~lna/ f~A~~~ Drprawnw ./ NwwMlsw ..f Food fiirwce. Mor.- ~ l.alilw of TecMoloRy. Gar l. EDWIN WOOD. M.D, I.a.on.r M sr. .o. U.i.aakr School .t SUMNER WOOD. l.. M.D. Asdwr ho/tas, of rrAa~~. Tk. lal+.. a~as. R~.Ri. ord.~. ~r Setad of MeD JOHN P. WYATT. M.D.. ho raaa o/ PatAdoq. S.iM LarM Univordly Sctted .( Medk/.e, S(. L.sria. KOJI YOQHINAOA, hrD Ceret for t•aratlMio. Rs.e.reh NIaD. Netb..l LNNwes o( Ik.MA. Merbed.. MO. 117

INDEX OF SENIOR AITTHORS Ahranrs. W. R.. ): Alhanu, W. A.. 2'_ AnJenwn. K.. 61 Anamiatk+. H. N, S 1 Arnnow, M. A , 56 Barrineau. L. 1.., 24 Ikany. K.. 29. 3' Brrmrern. 1.. R., 52 Bhanarharya. A.. 84 Branghkr. 1. M.. 65 Bruwn. 1. K.. 33 Bosbee. [). 1.., 66 Bush, 0. A.. 76 Carp. H.. 28 ('halrahar(i, S.. 7 ('harurvede. A. K.. 73 ('uahon, 1). W.. 16 ('oughlin. S. R., 49 Danes. B. S.. 20 Davns. B. D., 62 Diala, E. S.. 12, 14. 15 Downey, li. F.. 55 Ellis,1.. 61. 64 Erickson, C. K., 66 Foster. l. A.. 23. 25. 67 Foa• R. B.. 36 FrieJman. (i. D.• 99. 90 Galanakis. 1). K.. )S Gcokas, M. C.. 48 Uiekn,l. E.. 1 I Hantmer. R. E.. 70 Ilarada. R. N.. 40 llanwig. 1. H., 43 Ileqny, K. M.. 56 llo. M•K.. 96. 17 IloQman, R. M., 14. i S Ilolrnberg, L A.. 85 Ilwang, K. K.. 70 laiswal, R. K.. 74 lanoQ, A.. 30 Kaprio. 1., 91, 9), 94 .Karr. S. R., 67 Kci(h. 1. M.. 47. 49 Kostenvuo, M.. 92 Kouri, R. E., 10 Kremers. P., 12 Kurnner. K. F. , 17, 111.19 1 anginvameo, H .'N/, 72 1 aigntan. (' , 27 I ,+non K A , 14 I .1wrtn.r I (' , HO I,•,• W A1 1, 91 U, H. C.. 69 l.u, C. Y., 117. Mtt Lynch. H. T.. 21 Macher. B. A.. tl I Marks. M. 1., 61 Marsh.8, M. V., ti Manin. B. R., 60 Maeryama, K.. 43 Malheson, N. R., 31, 77 Oesch, F.. 72 Owee, A. 1. Ill. 30 Pagini. F. D.. 35 PaiReai, B., 10, 68 Panineta, M., 94 Pxdi, a. U., 19, 20 Ptalt, K. L., 71. 72 Powen,l. C.. 26.27 Rankin,l. A.. 82 Rasp, F. L., 36. )8 ReJdi. A. H.. 17 Rcilly, C. F.. 31 Rcpine. l. E., 37. 38. )9 Rhim, l. S.. 23 RuEni1:, 1. E., 13 Ryan,1. W.. 57 Ryan. U. S.. 58 Sstry, S. V. R., 75 Schapira, M.• 54. 33 Schuelke. G. S., 22 Ser.hen. H . 62 Shashy, D. M.. 39, 41,12 Snodgrau. D. R., 9 Snyder. D. S., 88 Springer. T. A.. 83.114 Stosul, T. P.. 45.46 Ta1e, R. M., 41 Taussig, L M., 47 Taveira Da Silva. A. M., 34 7 hompson.l. A.. 73 ToweB, 1. F.. 63.67 Travis,1.. 29 Vakriu., N. 11., 44 Van Canlforl,l., • Virca, G. 0.. 79 Wharlon,l.. 82 While. R., 25 Wiuina, R. C.. 719 Williams. 1.. T.. 53 Yachnin. S • 59 YamaJa. H., 3) Yamamoln, T , 79 Ym. 11 A. 44 Ymhuuura. 1 .27 t1N r V INDEX OF PRINCWAL INVESTIGATORS Acelo, M. D., 60 AnloniaJcs,11. N.. 49.50.51. 52. 53 Biswas, D. K.. 7 Braughkr, J. M.. 65 Busbee, D. L., tl, 9, 66 Cochrane, C. G., 78. 79 C'ollins, A. C.. 61 (•olman, R. W.. 54. 55 /)owney, H. F., 55 Erickson. C. K.. 66 Erwin, V. G., 63.67 Fouer, 1. A.. 2), 24, 2S, 67 Friedman. U. D., M9, 90 Fuse, K.. 61 Geukas, M. ('., 27, 48 Ges+ner, T.. 6E, 69 Giekn. l. E., i, 11, 12 GoW. W. M.. )3 1lanwah. P., 34, 33 Mary, C.1., 70 HoRmaw, R. M., 12.13.14. 1 Hojnacki. l. L. 36 llon, W., 62, 63. 64 laao/l, A.. 25. 21, 30. 35 Keith, 1. M., 411 Kouri. ft. E., 10 5. 1t Kueltner, K. E., 17.18.19.20 I.ajlha, A., 62 liwrence, E. C.. 80 Lioncni. F.1., 56 Lynch, H. T.. 20. 21. 22 Macher. B. A.. 81 Meier. H.. 23 Mitche8,1. A.. 70 Oesch, F.. 71, 72 Paiten, K.. 10 Peunen, 0. R., 73 Polak,l. M.. 82 Ran/as.b, 1., 90, 91, 92, 93.94 R,pi2ee,1. E., 36. 37. )t, 39, 40.41. ReynoWs, H. Y.. 82 Ryan, U. S., 37, Sta Sastry, 111. V. R., 73, 74, 75 Springer, T. A., !), 24. 83. 86. 27 Slossel, T. P.. 43, 44, 43. 46 Taussij L. M., 47 Travis, l., 26.27. 29. )i, 32. 76.77. 78 Unaue, E, R., 87. at Weinbwr. O.. 32. 33 Wi8,1.A.,47,4t Yachniw, S., 39 . . 119