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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