Tobacco Documents Online

agents which react with cellular macromolecules (DNA and/or RNA) is required Ior their carcinogenic or mutagenic activity; (2) In vi:ro studies suggest that nilros.minn are activated primarily (but not esclusively) by cytochrome P•ISO-dependent microsomal miaed-/unctioo osidases in the liver; (l) The postmicrosomal soluble eytosol also contains DMNdemeAylase ae- tivity, present in the "pH S e+wynse" fraclioe; (4) Thne miacd-/uoctioa osidasn are subject lo inducliom and repression by various agents and, thur, they may influence the bioactivaliow and eareinocenesis by nilroaamines Soene of lhese mndiAers may inAuetses uitroaaaaiaa earcinollenesis by mechanism(s) unrelated 1o the tnhancemenl or iahibilion of ailro..mine metabolism, e.g., iaMrenciag DNA mnformatiow, the eW o( DNA npair, etc. Lai, D. Y. and Arros, I. C. [i/r Sciewres 27 ( 2 )): 21,9.2165, 1990. From the Fnvironmenlal Protection Agency, O/liee of Toaic Substances. Wash- ington f) C, and the 1]rpartmeal of Mediciae, Tulane University Medical Ceoter, New Orleans. STlrp1FS ON MFTABOI.IC ACTIVATION OF CHEMICAIS FOR MAMMAI IAN ('F.LL TRANSFORMATION ANI) MUTAGENESIS There are numerous chemical agents in man i environment these days which have been implicated as potentially hwrdorn mu/akens and eareino- 6em This review paper louches upon several methods that have heen devel- oped to screen these atems lor Irwenbal problems Speedkally, during the las/ Afletn years numernus mammalian cell sywems have been devised which can detect such chemically induced endpoinas .s eytoroaicNy, /rans(ormatinn, and mulagenesis Ilowever, many chemicah demonstrate toaic or mutaeenie eRects only after they have becn metabolized by cellular enrymes to more reactive fcxms. O/ten the usefulness of a specific In virro assay system may be limited to only one or a very few classes of chemicals In order to extend the range of in rlrro nsays, various methods of wpplying exogenous melabolie activity have been tested. Cell-mediated adivNion, in riro-Iw vitro activation, liasat/orpn-mediated activaliou. and ocB.free enzyme preparations have beea used in atleatpls to duplicate the a.N.bolism which occurs during iw viro eipowne. 11 Is boped that N existing methods are perfected or new methods are di.covered by which to supply in rino tests with eaogenous metabolic aaiviy, krnwledge of the cowrpka i.rer.ctions which eveatually lead to tu- mors iw rlro wir sinurllaneously be iaereased. iltrstt., R. D., Kouri, R. E., ad ScisecAlmaa, L M. (Mkrokioloj*ic.i Auo- ci.rer) Is: h/i.hra, N., Kunkel. V., and MeMm.n, IN. (eds.): Adrawrer /n Modern Enrirowiwrar.l To.irobp, Vol. 1, Princefas lunctiou, N. I.:Seaate Prea lae, 1911, pp. 719-I37. OtAer aurr.rtr U. S. environmeMal Proteqiou Agency. From the (kpartmenl of Bicschemical Oneology, Microbiological Assoeiatn, 'Bethesd., Md . Ll1N(i CANCFR MODFI. SYSTF.M USINO )-METHYLCHOI.ANTHRENE IN INBRFD STRAINS OF MICE A lung cancer mndel system using intratracheal instiNatioe of 3-nse"- chol.nthrene in ClH/AnfCum and BCIFr/Cww atria.s of miee is presented here. The aniuah tested were free throughout their lifetime of Sewdai visvs, pneumonia virus of mire. and infedious disease. lu tltis study. 1the occwreacs of sponlaneous and chemically induced lu" eanocn was determined over the lifetime of the mice. Da1a were aaalyrsd by the acluurW wrethod for luas Iumor probabiliay, and /he probability was found to be dose a.d Nme de- pendent. Resuds showed Ihat over 95% of the l-etethykhd.wthrene-ureNei BC')F,/Cum and over et% of the Clll/Aa1Cum mice were found a/ death to have pulmonary eareinomas. Tunsors observed In aw)mah that died up b 40 weeks on lest were almost always squamow cell e.reLsomas, while tumon which were observed in aeimah 1hN died after SO weeks were mainly alveolar adenocareim>,nas. Both tsarqr types metaatasiad widely. Spoutaneous IuaB canecn (only alveolar adeaocarciaornas were observed) occurred in /Mese two strains at low frequency and were espaessed late in life. Thus, thn ays- Iem al/ords a suilabk model to study the indnctiow, eapresaiow. and progres- sion of luns tumors under cowditiorn where a vw majority of aairnds de- velop neoplasia. 11eery. C. l. rt .f. ( Mirrollolosk.l Asrorlarei ) Cancer Research 11: 5027-S012, 19s 1. From the Departments of E:perimental O.eology .ad Siochemical OncoloA, Microbiological Asaoeiata, •ethesda, Md. DNA REPLICATION AND UNSCHEDULED DNA SYNTHESIS IN LUNGS OF MICE EXPOSED TO CIOAREITE SMOKE A nmrine model system was used here b study the eRects of ehroole cigarette smoke exposure otn enzymatic acli.ities i. k+t.B liure traocided with DNA replication and DNA repair. For this work, attioe were chroukay a- posed to measured amounts of machiue-gewerated whole Kentucky re/en.na 2A1 cigarette smoke. DNA replication awd unscheduled DNA ayathais (UDS) were measured in hatB liswe in rino using a shorl.lerm organ cuNun method. Withiw one week of be6innieB stnolte etpeurre. DNA nplieati.+ aaivity, u indicNcd by incorporation of aH-lhymidiwe ialo total hruB DNA. was increased more thau 1wo-fold over sham<aposed eontroh awd rerwalued elevated as long as smoke eaposwe was continued. Trealsueut ot /unB tisrsa In vitro with either the hrnB carcinogen 1-aitroquiraiine-l-cisidt or asethpl- methane sulfonate stimulated UDS. presumably as the result of DNA repair activity, up until the 10dh to 121A week of sesoke e.posure. At that tima, however. the accumulated deposition of total particulate ma/erial In the lung was approximately 40 mg, and the level of UDS sdemlNed by the alkylatiag chemicals declined to approximately 50% of that seeu in lwy thwe lrosn sham«posed cootrol mice. If the mic% were removed from wnoke eapow.re. DNA replicalive activity returned to normal levels wilhia one week, bu1 the UDS response to DNA damage remained depressed up to Ilve months aller le 1 19

i ending smoke erposure. The results presented here show tlat both transient and apparently permanent changes are produced in mouse lung as the result of chronic exposure to cigarette smole. Rmrnwren. R. E. rt d. Cancrr RrrtaaAll(7):2St)-2Tlt, 1981. From the Dep.rtmeM of Cowrmrnit7 and Environmental Meiicine, Univenily of California. Irvine; and the Departments of Experimental Oncology and Biochemical (McoloDr, MicrobiobRical Aswciates, Sethesda, Md. MOSILITY OF SURFACE PROTEINS ON NORMAL RAT MACROPIIAUES AND ON A`MACROPHA(3E1-IKE' RAT TUMOR 'flThis study was MMlated so eaanni.e the lateral tnoMilNy of the same mem- brahe rnoleculcs is n..ctopha{ta, twrror cells resembling macrophaRts and lyrphocrtes. Results of these edl atrdies showed that peritoneal macrophaRes e.docytored thek histocornpNRrRiyr atwiRens (RTI). Fe receptors (FcR). and ooncawavatin A (Con A) dter tror-li.bing by IipndR, but did not cap these membrane proleins. 1. Nrrser teMs rese+nblinR eucrophaRes (721N), eheat membrane proteins were capped afler binding lipnd.. Frperiments a+earur- ing fhrorescence recovery after photobkaching showed that the mobik /rae- tinn of RTI was .iRnifkantly lircater In 72)N cells than In normal peritoneal macrophars Pmumal+y. the memMane praeins of 323N are not as kMered to the crtoslekton. or, 1/ .o, are in a nesus that is not the aame as that which occurs between membrane proteins of normal nucrophaRes and the eyto- stekbw. TT+e mobility of RTI and Con A receptors in normal lymphocytes was di/lerent from that found in tat.sal macrophaRes and imilar /o that of 721N eeMs. Thas@ observationa wyest that the wnov.emnl of maMbraM molecvks is determined by ocR type and ia regulated by the cytoskekton which varies Iw alrncture and functio./wm eeR type to cetl type. WoJ., a. A.. YRuerabide,l. and PeWtwatt,l. D. The lorrnJ o/ Crn ablon 9o:70s-71o, 19111. Other swpp.rtt National Iu.litMea of Health and the National Scie.a Pots.datio.. Prom tha Department of Ana/omk PathologT, City of Hope National Medical Ce+wer, Duarte, C.1.: Dep.rtmesA of Siotosy. University of Calirornia at San DieRo, Lt lotla• and the Dep.rttneat of Immuoopatlwlop. Research Institute of Scrippa C11nie, La 1oMa. VENEREAL SPIROCIIETO.SIS OF RABBITS: DESCRIPTION AND DIAONOSi3 Venered spirochetoais of rabbits (VSR ), a disease caused by Trrpewrmw rrwkri9. produces ksiona thN are characterized a papules. ulcen, or crusty areas. In the study of VSR reported here, rabbits nsed were from the /nbred, partially inMrd and mutant hearing strains maintained in conventional and hr.orrrcr.wnr •krr.rd c.r.w.rs at the Lclson I ahorarory. Ilar Narhor, Me Rea.tt. cho-rd rtw ..rur.llr and e.perrmesralty induced genital ksKrns were :o I similar clinically and hi.to/cKicaN!/. The rwrrl common .ite of lesbro b aal urally infected rabbits was the v ulva or preywree. Other sites, in descendin, order of freQuencp, were anal reRion, .oae, e*rJid, and lip. Clink.al appearanc, of ksions varied from erythematow nraeulea .r papules to ero.ionr, ukers anJ crush. IlistoloRical e.amination of k.siona cortflrmed the pre:senct of cro sions, ukerr, and erusts and, In addilion, reveded acanthosia and I.filtratinn o the dermis by plascna ce11k and macrophases. SirK+e diagnosia tl/ VSR de penda upon fieding T. crn(n/1 in wspect ksiantr or demonslraiing porNiw serological leus, four methods were used to test 11rc k.ines. Dark Rdd eenr ination of scrapinp /ronr lesions was auperior M the other anetholr 1or dr teelion of Tirpowrnv crwlrnll. However, the rapid plasnu reaRin hrt desedev' more Infected ralrbita than earnlnation of ksions. The freq uency of lalr posi tive rapid plasma reaRin lesta was low; the frequency of /rrhe neplire ara/in tere varied with the slqe of the diaeare. Cr.fde-Ikamer. T. 1.L and Foa, t. R. (Me/er, H.) l.6o.do.T Awl.wd ScJrncr )1(1) : 766-771. 198 1. Otfer.rppe.tr Natiend IartMwes of HeaM and The lactw. Laboratory. Prom Tlie lactson Lsboratory, Bar Harbnr, Me. VENEREAL SPIROCHETOSIS OP RABBITS: EPIZOOTIOLOOY The role of vertical tr.nrnis.iot and Renetie predisposition i. 1M qirad tioloRY of venereal spirochelosh of rabbita was evaluated In this study. Rad bia surveyed here were /rern the i.bred,p.n i.Ry inbred and Otwatd Marly raralm In ths evwvaMion.l (etstoa/kaRp Wae1N) and hyrlereAa>Mla/ved colnnies maintained at TM lactaew L.borNer7, Rar H.rbar, Me. R.wRr presented in this p.pa sho.r diRerenoes In the rrneepibility ro and e.prerriom of venereal spirocheto.is assortR diRereM iobted or part i.bred nraitr of rabbits. While aignificaM diQeretsces het.tea Ireauene~ lesions M aduM twale or fenuk rabbin were no1 observed, nor were aiRniAeaM diilereass betsrcen the frequency olletiota is breeden or pe.breeders i. aht ti-/2 r.aath age tirarP. aigeilkaM diRetreoes between die freryttencp of kaionr In brecden and won-breeden wero oMer.ed srhew dilkrenl aRes wete pooled. 71w fro- quesrcy of venered .piroehe/osia in rabbib kaa Ihaw i snonths of age wa bwer tham thN in adtrlt rabbits. Newborn toMering eaperirne.ts indkNtd that infection occurred at birtb and during the tnrctNng period. Evabation af Ih.- terectomy-derived oRspinR of inktted datns suggested venereal spirocf,ero.ir was eliminated by hpsteredareyderivatiow. Vewerea) apiroehewrM was we- cessru8y aransmitted by topical or intr.derrnal suhesN.eeau at" Lowlaiou of adult rabbits. Adult rabbiy were wsore twceptiMe to eaperinrcs/a1 reneual spirochelosis than nconates. These eaFhuiorn were based on clinical a.d aeroloRical obsenratiotr. CunGAe•eeamer, T. L. and Foa, R. R. (Alrkr, H.) l.boraro.r A nbnat Srlrnre )/( d):)72-77R, 198 1. OtAer aupport: National Institutes of HeaMh ard The lactso. Laborasor7. From 7he lactson l.abcxalory, Bar Harbor, Me. 71

I VENEREAL SPIROC1113TOSIS OF RABBITS: ERADICATION This study wu undertatten to (1) evaluate the clinical am1 serolo6ical responses ta therapy of rabbits infected with venereal spirochetous, and (2) develop a program to eradicate venereal spirochetosis from an enzooticaRy Infected eanventional rabbit eolony. This venereal spirochetosir of rahbits, caused by Treponrw+a crnkrif, has been observed since the 19401 in the rab- bit colony maintained at Tbe laekso. Laboratory. Treatment rrt Individual rabbits with obvious clinical lesions rsing a produd containing crystalline prnkitlin and dihydrostreplorn7drt esred ksiona to hed, but venereal spiro- chelosis persisted 1n the colony. To further observe the eAects of healing here. clinical and serolo6kal tespowra of rabbib receiving three subrutaneous M- jectiona of bernuhiee penicillin O-ptocalna peniciYi. O(eirtw r 42.000 or 11,000 IU/kg body weight/week) at sevew-day Inter.ab wert reo.itored. Both doscs were eAedive. Tlre lesions healed wlthin two weeks of tte Ant treat- meM and rapid plasma reyin tilers declined markedly or disapftared by the .i.th week after the Ars1 treMnteN. Based upon the abovt Anc'inRs and re- suks of previan epaootioloRkal studies„ this program to eradi:ate venereal .pirochetoais lrorn the enrootkaRy infected colony was successfultj undertaken. (1rdN1e-Bea.rer. T. L and Foa, R. R. (Mein, H.) lAAoc.rery Awln.J Scirnre 31(1):179-3t11, 1981. Oth.r.upr.rtt National lrwltules of Hedtft and The lackson Lrboratory. From Ttr lacfson Laboratory. Bar Harbor, Me. CHARACTERIIJITION OF MOUSE FETAL LUNG CELLS CUI-TURED ON A PIOSKIN SUBSTRATE 1. Ihe methodoloRy, study presented here, an attempt was made to Rrow orpnotypic tnouse fetal lung cultures on pigskin dermis and to identify th. nxsu8aat oeM ekments by morpholoRial and histochemkal evrdence. Speet- AcaYy. lung organ bits wete Iaken for this study from fult-term mice and esplanted on the dermal surface of sltrik, dead pigskin. The cells mitirated onto the pigskin dermis .nd ptoliferded to form as organoid culture eonaist- ins of ductular slnrctures separated by a matria of epithelial cells. Cells within tha ductular structures were ciBated, produced mucin and eshibited the ac- tivities of nonspeeiAc eMerae and pmma-6/u1-amyl transferase• theretore. Ihey wert considered /o be derived from bronchial epithelium. Cells forming the nntrin poaseased the activities of nonspeei0c esterase and alkaline phosphatase and contained IanteRar strudures typical of wrfactant•producint pneumocrte Type (1 cells. Bec.use of these propertie+. the cells forming the matria were eonsidered to be derived from alveolar precursor cells. Overan, it seems at this time that this pigskin culture spnem, which cornbines elements of organ. tiasue. aed cell cuhure, will probably prove to have direct applicatinn In studies of rnutqe.esis. carcaa6enesis, and cell-to-cell interaction. Yo.hid.. Y cr.l (Frnn..w. A E 1 In Vlrro 16111 411-W1. 19110 From the I a lolls ('.ncer Research Foundation. I.a loila, Cal. 22 I . FINF STRII('TURAI. InFNTIFICATION OF OR(:ANOII) MOUSE LUNO CEI.I S('UI TUR FI) ON A PIGSKIN SUBSTR A1 F. This paper, which reports primarily on Ihe Ane structure of fetal lung esplanls and cwntrowth on gig.kin dermis, contains an initial report of a new cell type intermediate hNween ciliated cells and pneumocyte Type 11 cells. This cell 1ype, not yet found in vivo, is characterized by both cilia and lamellar bodies. In the overall body of work presented here, eRplanKd organ bits grew on the surface of, and into, pigskin dermal collagen for at kasl nine weeks after the inNiatinn of culture. The outgrowth consided of a thick cellular sheel containing variorn sizes of ductular strucNKes within a cellular matria that did nnl show any particular strudure. Electron microscopic observation revealed that the larger duclular unrcurres consisted Iar6ely of ciliated cells. The smaller ductular alnrcture eonsisted lareely, of Type 11 pneunsocptes eon- laininR lamellar bodies. Aho, when Iht cellular nulrit was eaamined, it was shown to cnn.ist of Type 11 pneun.ocytes and other cell types including Abro- Mrrs and macrot.hadcs in the early salle of cultivation. MacrophaRes invaded the pi6skin dermal cdlaRerr. An iMermediale cell lype, mentioned above, was idcnliAed in the larper ductular structures. Upon comparison of the uNra- slruclure of the or6araid 6n vitro cultures in pigskin with the original fetal lung. il was apparent that cylodiQerenlialion had oeeurred. The orpnold eomponents and strocture of the euhured cells nrore closely resembled adult long than the fetal lung use& 1o initiak the cultures. Yoshida, Y. Hilborn. V. and Frcewr.w, A. E. IwYl.o 16(/1):994•1006, 19/0. Oth.r wrprtr Awteriean Cancer Society and the National Cancer fnstitul.. From the 1A lolla Cancer Research Foundatien, L. lolla, Cal. OALACTOSYL TRANSFERASE ACTIVI'iY IN RAT BLADDER TRANSITIONAI. CELL CARCINOMA LINES AND IN EXFOLIATED CELLS IN URINE DURING CARCINOQENESIS AND REVERSIBLE HYPERPLASIA This paper describes the properties of plactosyl Iraasfense (OT) ho- lated from rat bladder transitional cell eareinonn in cuRure, and investiRMa the enzyme's activity iw (1) esfoliNed bladder epitbelial cells in urint of rab during carcinopenesis and rcverar~le hyperplasia, (2) bladder lutnor eella and (1) seria8y transplanted rat Madder eareinoma cells. For this study, cuhured cells of tat bladder transitional cell careinoma line AY-27, in suspen.io., were assayed for OT br measurement of the transfer of /sll/ galactose to de.ialyded ovine submasillary mucin (OSM-hANA). The asay was optimized witb re- apect to time and to protear, wid'arc diphosphale lodadose. OSM•NANA and Triton X-100 concentrations. Thia assay was then applied weekly to suspe.- sions of esfoliated Wadder cells collected from wMn of rats fed the Madd.r carcinogen N-(1•(3-nitro-2-furyl)•±•thiazolpl) formnwlde. .nd of oaMrd nb. Results showed that increases in activity over controls appeared 42 weeks after feeding the carcinogen. N a stage when bladder tunwrs were already mkroinvasive or deeply invasive, and activities at 32 weeks were about BO- 23

1 fold greater than normal values. In contrasl, a bladder cytolosic agent induc. ing revcrsibk hylrtplasia was Injected into rats and ea(olialed cells were eolkcted froen urines: these cells showed no greater GT activity than normal. Bladder tumor rissue from a transplanted tumor had the same high specific enzymatic U T activity as eafoliattd cells from tumor-bearing ra1s. Plottin, O. M.. (Zilbert, S. L., Wides, R. 1., Wol/. Q., Cohen, S M., and Futushima, S. Cenrrr siorAemtury eiopAyrkz 1:251-236, 1980. Othe..rrpertt U. S. Public Hea/th Ser.ica. From the Department of Nutrition and Food Science. Massachusetts Institute of Technoiogy. Cambridise, and she Depattwrenl of Patholosly, St. Vincent I/ospital and University of Massachusetts Medical Schod, Worcester. T11E ISOLATION AND CHARACTERIZATION IN VITRO OF NORMAL FPITHFI.IAL CELLS. ENbOTHELIAL CELLS AND F18ROSLASTS FROM RAT URINARY BLADDER Since the physiologic MNeractiow which underlie epithelial-mesenchymal inlegriry seem central so she understanding of tumor invasion, an attempt was made in this study so isd.ue and characterize in culture she normal eclb- lar elements which comprise she e.dolhelial-stromal junclion. For this rea aoe, epithelial celh, microv.scular endolhehal cells and fibroblasls were iso- laled in culture from normal urinary bladders o( Fischer rats. T1These ceR types were rtlSased from ewzymatiall7 ditaled, evertcd urinary bladders. Their cultivation by standard lechniqrrcs yielded fibroldasloid or epithelr.id nwno- layen. Cultswe growth characteriMics and light microscopic eaaminations readily idenlified librohlasts as the source of Abroblastoid nronolayers. How- ever, tbe.e techniques were swbk to diQerentiate belween she epithelial or cndo/helial origin of epithebid eronolayers. Electron microacopic examinations were successfully applied is sonar eulluret~, bait failed in others. The lhree cell types studied hers represent she normal control eeNs of an in vitro twnor aa)del /or she study of i.vasiMetleat. AII drra cell types are Involved in the formation and functional maiMera.oe of the epithdial-uronnal junction. The study of cell-cell and eeR-matrat interaclioas may provide (mportanm clues for the understanding of twnor invasivenesy a process that starts at the epithelial- stroval juwctioo and proceeds with iu destruction. PwR, 2. U.. A.Aersor.. S. N., Memoli, V. A., and K.rrtnrr. K. E. T4iw f CeM 12(3): 119-1)2. 1980. Othor er' prt: National Institutes of Health and the Otho S. A. Sprague Memorial Institute. From the Ikpartmenb of Pathobgy, Orthopedic Surgery and Biochemistry. Rush Medical College and Ruah-Presbyterian St. Lute's Medical ('enter, ( -hic ago. 24 11. The Reopir.toPy Sy.tewa PULMONARY FUNCTION IN YOUNG CIIILDREN WITH ALPHAr- ANTITRYPSIN DEFICIENCY. COMPARISON WITH MATCHED CONTROL SUBJECTS In this prospective study, designed to see it alph.,-antitryp.in (AAT) deficiency leads lo IwtR Impairment in early ebilahood, the peboonary /ww tion of a group of 19 children wish moderately severe and severe AAT de- fkiency (Pi phenotype ZZ or S2:) was oornp.red witb that of healthy oontsok matched for age (l-7 yean), se: and sise. There were .o statistically sifi- nilkanM differences between the /wo poups in the three pulmonary /wrctioi, variabks studied. that b, functioeal residual capacitr (FRC). maximal ea- piralory Ao.r at /'RC (Vma...,) and aineeotreeted Vaart„. (Vrnaz,.r! FRC). On the avera6e, however, the AAT4eficknl cbildrew bad higher FRC vahus and lower FRC (Vma.,.r) Ihan the eontroh, both of which would be expected i/ these children had some measure o/ Mepaired pulmonary fu.c- lioe and air-fiow obstruction. Although this strdy does not provide a deW- live anawer, il does suggest thal ther. Is no Rroea hnpairmenl of overall pu{- anoaary lunction in AAT-de/kient children under aee seven. Raii.t, A. S. et al. AnrrrJr.n Review o/ RrrplraorP Dluase 122(6):617-622, 1980. Frorn the Departments of Medicine a.d Pbpsiolop. University ot Ore6o. HeaNb Sciences Centu, ParW.d. REFERENCE VALUES FOR FUNCTIONAL RESIDUAL CAL•ACITY AND MAXIMAL EXPIRATORY FLOW IN YOUNG CHILDREN Tlre assea.meel of pubnonary fsactios M yowtR ebildres has always bees dillkdl, but a wesw lechniqre th.t tae.sures swMd eapkatory Aow a fu.e- tion.l reaidr.l capacity (Vnua,,,) M eaajwetiow with . wteawrerwent .f functional residual capacity (FRC) ia .ow being i.vestipted for ks suitaW/Rp n a rwcaswinp tool. The objective of the swdT psesesNCd heee was 1o deter- aaine the relationship of FRC and Vmaa,.e to sea, aie, beiRtw, .nd .rei61N in a group of heahhy young childrel is Port4.d. Oreron, and to t+sa these meaurements as reference values M laler sludies. Within the aRe, heiRlM awd weight ranges studied, exponential or multiple regression techniques offered .o substanlial advantage over simple li.ea regression YsInR heitlN, weiRM or qe. lAcre were no sex differences /or the relationship between either v.riable, and ate. heigM or wei&. These techniques eaw readily be wed in cbildren as young n three years of age and may provide a method for studying lung Sro..tb and development in carty childhood and a way lo observe the propes- sion of disease or the effect of IrealmeM in Ilte young child. A.rLt, A. S. rt of. Anrrrir.n Rerlew o/ Rr.plrnmory fNuarr 122(6):9e)-9ts, 19/10_ 2S

Qtlrrr support: National Ileart, Lung and Blood Inslitute. From the Departments of Medicine and Physiolody. Uni.ersiiy of Oregon lledrh Sciences Ccnter, Portland. Ct.FARANCF- AND METABOLJSM OF CIRCULATINO PANCRP_AT1C PROF.t.ASfASE IN T11E RAT The rate of clearance ol eircylNiwg rat p.ncreatie proesaatase, as well as the metabolic tale of this protdn, was eaamined in this rat madel study. Reaults presented here showed sh.l dte clearance of 's'1-lahekd proelastaae was biphask, with a hal/•life for ckarawce of free 27.000-dalton proelastase of app.ouimately 7.10 n.inutes; a slow eonrponent of clearance possibly due to proelastase associated with plasma protease inhibilors was also observed. At 10 minutes after injection of 's'1-labekd proelastae into the circulation, the major fraction of the i°I wu (ound to be locdized in the kidney. How- ever, appearance of 's91 M wine was sio., wilt 16 houn being required for e.cretiow of 50% of the i.jected 's'1 as acid-sohrb/e tnalerial. Suhcellular k+calizatiow eaperiwtcwts oft hornogenates of kidneys renrovcd at 10 miwu>tes pauuinjeetiow revealed that the majority of the 's'1-labekd tnaterial was aaso- cured with the nti/ochondrial and lyso.orwd fraction. SucroK kradient eeMri- fugation studies of this Iractiow showed that the labeled material passes from Ihe membrane Iraction lo the lysosomal Iractioe with time. In summary, the results of these studies demonstrak that the kidney ia the major site of ekar- anu and catabolism of circulatiaR p..creuie proelaslaae. Lsualenan, C.. Ray, S. B, Brodriek, l. W, and Crotat. U. C. Antrrkan lour+.1 o/ rAyriobp 279 (Oastrointest. Liver Phyaiol. 2):OS0d• 0510.1960. (NAer eorrortt Medical Research Service of the Veterans Administration and the National Institutes of Health. Froet the Fatzymoloky Research Laboratory, Mutinez Veterana Adminiatr6- tio. Medical Center, Martine:, Cal.; and the Department of Internal Medidee, U.iversity of California School of Medicine. Davis. CLEARANCE OF CIRCULAT1NO ANIONIC AND CATIONa: PANCREATIC TRYPSINOOENS IN THE RAT In this atteetpt to gain insidtt iMO the physiological trtechanisms eon- trolliei the levels of pancreatic trypsiwotetr in human blood, the kinetics and mechanism of clearance of patcreNie eatioeie and aniosic trypsinoRene from the circulation were investigated ie a rat model. Eapcrintental proced- ures used here included protein iodination, plasma ekarance determinalion. Sel flliratioe study, and orga• distribution analysis. Results of these invesli- puona show that 's'I labeled rat calioeic tryp.inoges is cksred from the i bloodstream with a h.lf-life of approaimqely asti minutes. In contrast, 's'I- labekd rat anionic srypsinoden has a hall-/ife ie the circulation of appros- imately 55-60 minutes. Neither zynto6en binds to plasnu proteins to a aig- silkaM eatent over a period of three hall-lives. The relative ralea of ckara.oa of these zyrttogero Irom the circulation appear to correlate with their respec- tive isoelectric points. Praearily, the results preseated here show that the kid- aKy is the primary .ite of clearance /or both rat anionic and eatioeic pa.- ereatic trypsisoges. Brodrick, ). W., Larprtan, C., Ceoi.s, M. C., O'Rourke, M., a.d Ray, S. B. Arncrk.n Jorrn.l o/ rlirslolopr 239 (O.atroiaest. Liver Plyaiol. 2):aS11- OS1S, 1910. 01Asr wr'ort: Medical Research Service of t6a Veterans Adntioiatration and the National lnuitutea of Ileahk From the EnzyrnoloRy Research Laboratory, Martitte: Veterus Adrsi.irr.- lio. Medical Ceuer, Martinez, Cd.; a.d the Departaney of Internal Med1- eioe, University of California Scbool of Mettieia, Davi.. SECRETION OF ALPHA.-PROTEINASE INHIBITOR BY CULTURRD RAT ALVEOLAR MACROPHAOES Alpba,-proteiaw inhibitor (.,Pi) iM thotspt widely 1o be tb tta.jar esdo6eaous inhibitor of kutoere elaslne. 1n dr eaperitwew pr+eseae+ 6ere, .,Pi .ecretio. (row alveolar wtacroph.Bcs was eznniaed, ber,at..M o( .,Pia k.osrs role ia proteiuse-aMipraei.a.e teRulation a.d tha polentl.l importasoe of its loeal prodttctien by hang oer.. Speciticaqr, .,Pi, tuata- bolicaly labeled with a'S, was delected i. coaxurated euNure aed'turs af rat alveolar wucropbaRes by itnaNrodeelropAoresq .woradioRrap6y. Tke otetabolieally labeled macrophq. .,Pi 6ad t6e eraeo deetropiosaie molsiWy as did pure rat serum .,Pi. Moreover, wtowotpecilic atuiserunt 1o rat sasm. .,Pi precipitated a a'S-/abekd protein froat wacroplt.ige culture wperetalM asd /6is proleia waa ahoww by sodium dodeerl attYate polractylamide ild ekctrophoretis to have a ntokcWar weisht elo.. /o that of rat mtuw .,PI. The addition to the cultures of erclohetitttide at a ooaoe.tration of 0.30 p=/tnl prevented the i.corporalion of label iMO iatnw.oprecnbN material. Overall, the results presented here iediule ehat .,Pi ib syahesima atd aa crrted by cultured rat alveolar tat.crophaRes. It also appears trat the socretion of .,Pi by alveolar macroph.Rea might plar a rok is the defense of the Mt.R. White, R., Lee, D., ILbicM, O. S., sad l.no0, A. Atrterlc.n Revkw o/ Respiratory DJsease 127(1):4/7-41l, 19t1. Oth.r sr'prtr National Heart, Lung aed Blood 6ptNute and National Imtilule on Agiog. From the Department of Pathology. State University of New York at Sto.y Brook, Stoar Brook. 27 26

INAY-IIVAl1ON OF AI.PIIA, PROAFINASF INIIINITOR ANI) bNON('111A1. Mlt(YwS PROII:INASN INIIIBII()R BY ('1(;ARl.1'f E SMOKI. IN Yl f RU ANI) IN YfYO The eharacterissic destruction of alveolar walts in pulnansry cmphysema is believed to he dae to uninhihild activity of elas/ases derived Irom p.dy- mnrphonuckar Icukocytes and alveolar macrophars; and this uninhibited tnryme activity is helieverl to oeeur because of an imbalance bctwecn elastase and elaslase inhibitors in the lua>t. Many studies (both /n vitro and in vivo) have been done hy now on the protkrae/uNiprolease balance in the hrng, and the present paper carefully reviews /hewt. Sonre of the read,s pres.nted in this review show that ,4ueous solutions of unfractionalcd cigarette smoke urppress compka formation between hunran alpha,-proreinase inhibitor (w,Pi) and clasuse In vhro. OaidirinA qerws reproduce the effect of smoke, while snti-o.idanls prcvenl it. Inhalation of eiprette smoke in rats causes a siB- ni8eam decrease in functional activity of rat lung w,Pi Iw .•lro, which can he restored by trealmenl with reducing alients. Human bronchial mucous proaeinaae inhibitor (B%IPi) is aho inactivared by cigarette smoke and osi- danes in vitro. Functional aciivity of BMPi obtained from h.tman smokers is 20% lower than that from nonsmoken. Theae results support apresenled hypothesis that local inactivation of anliprottases in the reapirslory units and conducting airwaya of the lung by inhaled cigarette smokt might play a role in the pathogenesis oM chronic nhuruclive lung disease in man Lwo/l. A . Carp. FI and I ee. 1) K. Mr.Mr/in /wropwn ./r Phrunp+uA.due.r R..rr••nmrr 16(arjK+ /19F.0. Other aNp)rorr: National Ncan, I unt and Blood In.brute. From the hepariment of ParMrlory. Healrh Sc.ences ('enter. Slare 1lniversity of New York at Sp.ey brrrrk. Stony broark INIFRACl1ON OF CH1'MOTRYPSINO(iF.NS WITH a,•PROTI'ASF. INHIBI FOR In Ihis study, bovine chymotrypsin A (CTGN) waa used for the purpose of quanlitalinA the reactN.na of tynabem with w,-prMeinase inhihitors Results presented here shnw that CTON reacts with .,-Pt in essentially the same manner as Mwnan praclaslase 2; I.r., slowly to furrn a cernpka that is stat+k to denaturation with s.sdium dndeeyl sul/ale and A•mercaplnNhanol. lhe rate of ct'mpks formation was measured here by two mclhorH that were deviacd to determine the seenrd-ordct tate constant for the rexlinn of ('1(3N with w, Pl. In the lird. the rale of eompkn formation was detcrmined by monitoring the loss of thc inhihilor activity of w,•PI as a/uncrion of time. In the aecnnd procedure, the reaction of /harescein iwlhiocyanale labeled chyrootrypsinolen A with w,•PI was measured by Marescence p+lariralion. lhe rate of compka formatinn was - 10' of that measured for the reaction .of M.vine chym.wryptin with w,-PI. Dissociation of the compka was mN ob- served after dolulMn or the addition of e.cess bovine w-ehymnlryprin. As l.wlcrd by au.luun dnJccyl ad/Ne•pr.lyacryl.mide gel eketsa+phMean esperi• men@s, hunun .h)nnuuyp.m.1Krns I and 11 react with n, Pl at rates that art i appro.imalely equivalent to that determined for bovine chyrnotrypslaolien A. In comrast, bovine trypsinoten reacts very slowly with w,-PI, at a rale that is at moat tOs of that of bovine chytnolrypsinogen A. 'Tlrese results suggest that :ymogerr react with .,-P/ by virtue of partially formed active sites and that the potential active-site specificity of the :ymotee in part deter- mines the rale of xompka formatiow. Srodrick, 1. W.. Glsser, C. S., Larpean, C., Geoiar, AI. C.. Orauflo, M., Fassett, M., and Maeda. H. sioeArrnlury N( 2 t):1l6S-dt70, 1930. OOther wppertr Mldical Research Service of the Veterans AdministrNio., National Institutes of Heahh, Internation.l Union apNnt Canecr, awA the Ministry of F.ducatiow, Scknce .wd Culture of lapaw. From the F.n:ymoloRy Research Labo.atoeY, Manine: Veterans Admi.Wra- tion Medical Center, Martine:, Cal.• the Department of Internal Medicine. University of California School of Medicint:. Davis, aad the Institules of Med- ical Scie.cn, S.n Francisco. KINETICS OF ASSOCIATION OF SERINE PROTEINASES WITH NATIVE AND OXIDIZED a-1-PROTEINASE INHIBITOR AND a-1-ANTICI/ Y MOTRYPSIN In thb attempt b p{w an Msibht Into the physiofobkal function of tY two Inhibitors. .-1-proteinase Inhibitor (,r-1-PI) and rl-antichy.wtrrpsb (w-I-Achy), the specificity of each was investipted by sneasssrinB fho taN constuws of associaios with different proteiwa.es. Reported here. tbati an the sssocialiow rate constants for Me interactiow of rl-Achy, .-1-PI ad eal- diud rl-PI with several nwewaflas aeri.e proteinases. The results Indkw IhN kukocyte e/aalast reacts more rapidly with w-1-PI than any other ps- kinae tested, while ksMkoeytu caMepsiw 0 shows the strongest aswciNioM with r1-Achy. O.idatitrn of the critical .rethioeine residut: of redtroo ehe associatiow with krsdocyte ehntase by a factor of teore than 2000 ad also katrlrs the assaeiation with aN of the other enzymes ksled wMh the ea- eepiow of chynsotrypsia. SiteiAcanlly. oaida/iow eompletely abolishts a.J interaqion of .-1•P/ with porcine elastase, human plasmiw or hurwaw tAro.:- bin. 7Lese data support previous rewlu which iwAieNed that osidatioa of humae .-1-PI /n riro could rtdace the eRecti.eness of this hnhibitor Iw Qos- trollinB proteolysis. In the henB, in panierlar, oaidiiinB aBents of both chnn- ical atd biological sowces could Indirectly wpmeM elntolysis i. this tMwre, resulting in the dtveloporcnt of pulnww.ry emphysema. Se,Ity, K.. Bkth, J. and Tr.rlH. l. a the /orrnrf o/ elotopcd C/iewJiny 2SS(9):)971-)911, 19s0. OtAer support: National Institules of Health aad the Institut National d. la SantE tt de Ia Recherche MEdicak. From the nepartment of Biochemistry. University of (korala. Atheas; a.d the Facullb de Pharmacie, I aboratoire d'ErittirrrwbAie. U.iveni/1 Loui. Par teur, Strasbourg. France. 29 ?N

RF.AC71VtTY OF IIUMAN I.F.UKOCYTP. ELASTASE AND PORf'INE PAN(-RFATIC FI.ASTASE TOWARD PEPTIDE 1-NITROANII.If1fS CONTAINING MOI)EL 1)FSMOSINE RESIDUES F.VIDIiNCE T: IAT IIUMAN I.FUKOCY fF FLASfASE IS SF-I-ECTIVE FOR ('ROSS-1.INKED REGIONS OF EI.ASTIN Plastin, a/ksibk, highly cross-linked protein, is found in high quaMAies in mammalian lung and arteries. While the chemical structure of elastin is not ye1 tnawn, il hn heen shown IhN il contains a number of cross-linking amino acid residues such as destwosine and Isodesmosine which are primarily hydrophobic in character, but haw a positively charged pyridinium ring. These cross-linking residues arc /ornsed by the action of lysy) osidase upon Lys residues in tropoelastin, a preeursor of tlastin. Iro this study, a series of ktrapeptide "troanilides which eoddn Lys and a series of mod.fkd lysine residues were synthesized. The nadi/led lysiwe residues have various charae- teristks of desnsosine and isodesrno.{ne re.sidues, such as positive charge, a hydrophobic arnmatic ring. or a pyridinw ring. The reactivity of the ectra- peptide 4 nilro.nilides containing the model desmosine residues at P,. P,. or P, with human kukocyte (Ill.) and porcine pancreatic (PP) elastase was measured al pH 7 5 and 2S'C. Hl. elaslase eshibiled high reactivity toward the substrates with P, or P, hydrophobic proups. and MtO-Suc-I.ys(Pie)- Ala Pro-Val-NA was shown 1o be seven times more reactive that the previous hest Ifl. elstase substrate. The major change occurred in K„ values. The substrates containing l.ys residues wert either nonreactive or poor. Escept for two substrates with P, hydrophobic nesidvn. PP elaslase was less reactive toward the substrates containing model desmosine residues than toward MeO- Suc-AI. Ala Pro V.I NA The rnu/u presented here support the hypothesis that H1L elastne ckaves elaston selectively near crow-linking residues. Ynutake. A. and Powen, l. C. (Traris, J.) aiocllernirrry 20(1 l):)67 f-)679, 1981. From the School of Chemislry. Oeorgia lnslilute of Techoology, Atlanta. SUl1..ST1tATE SPECIFICITY OF 1WO CHYMOTRYPSIN-LIKE- PROTEASES FROM RAT MAST CELLS. STUDIES WITII PEPTIDE 1-NITROAN1l./DPS AND COMPARISON WITII CATHEPSIN 0 A kitsetk Mudy of the hydrolysis of a series of peptide 1-nitroaailides by the proteases from (1) typical masl oell., rat mast cell proteaae 1( RM('P 1), (2) pypic.l mast cclls, rat masl cell prottse 11 (RMCP 11), and human cathepsin O is reported here. eah rat enslmes are effective proleases toward such suMarates.- Suc-Phe-Pro-Phe-NA and Suc-Phe-Leu-Phe-NA are the hnt suhstrates for both RMCP I and RMCP 11. In the case of RMCP 11, both the P, (Phe) and the P, (suocinyl (Suc)1 groups are important. Suc-Phe- Pro-Phe-NA is the best • nilroanilide substrate yet for eathepsin 0. Observed suhsite preferences Indicate that the S, and S, suhsiln of RMCP I. RMCP 11. and cathepsin may he very similar. The K.,,/K„ values of RMCP 1/ and cathet,•.n arr soandkantly Mr.rrr than those obscrved with bovine chymotryp• cm RM/ P I .nJ ,rher v..nr p.owraws It u not ck.r .hether this diQerence i is related to their phy.iolos ical function or iw due to a non-optimum substrate structure. Prolyl residues at the P_ potilion enhance the effectiveness of wb- strates and inhibitors for RMCP 1, RMCP 11, and ea/hepsin 0. Unlike other serine proteases, both rat enzymes can also accept Pro residues in position P- of substrales. A peptide chioromelhyl ketone. Suc-Pro-l.eu-PheCh,C1, was synthesized in order to Iake advantage of Ihis phenomenon. This compound is an effective inhibitor of RMCP 1; RMCP I is inhibiled 10-93 timea t.rer than RMCP 11, cathepsin 0. or bovine chymotrypzin. This Inhibitor might be usefsd in elucidating the physiological function of RMCP I and RMCP 11. Yoahida, N. tr al. (T..vlt, 1.) siorAemis/.y 19(23):5799-310/, 1950. Other anrport: National Institutes of Health. From the School of ('Aemistry, (ieor/ia lnstitWe of Technolop. Atlanta, and the DeparlmeM of Siichemistry. University of Washinglon, Seattle. HUMAN PANCREATIC PROELASTASE 2 SEQUENCE OF THH ACTIVATION PEPTIDE Ikcause elastolytk enzymes are thougM to eause the destruction of elaar Iic tissue observed in atherosclerosis and is emphysenu, it seemed impoeta.t to scrutinize the actions of proelauase 2, an enzyme that has already hetw shown 1o be present in human serum. For this reason, work was dons es human pancreatic proelasta.e 2 and, in the paper presented here, the stnce ture of the activation peptide of proelastaae 2 was elescribed, and the IAew- tity of the amino acid residue at which cleavage occurs during activatkst. In vitro was established. In Ihis study, the N-lenrtinal aiaees residua of the amino acid sequessce of reduced and alkyfated bumaw pancreatic proelaMre 2 were established, and the N-lerwtinal amino acid residue was dhoww to be earbosymethyk:ysteine. A peplide containing an amino acid sequence e:orte- sponding to the Arsl twelve residues of proelastase 2 was isdated following activation of proelastase 2 with trypsin, perfonnie acid osidation a.d pd /iltration. Ihis peptide was not released prior 1o perfonnic acid osidation, suggesting that it remains attached 1o the major peptide chain via a disulAde bond containing the N-terminal hdf-cystekwe M a mrwter similar to that fosmd for chymotrypsinogem. The sequence analysis presented here suyesh dul human pancreatic proelastase 2 is more closely related to the chymolryp.iwolicn family than to porcine pancreatic proelastase 1. Thae resuUs clearly abw, however, that Mrntan proelastase 2 is a d'n1inN symolee. LuCnan. C., llrodrick. !. W. and Gro4rs, M. C. siorAln.ke er QiooAyric. Acr.63.f:20>1-212, 1980. Otber aupprtr Medical Research Service of the Veterans Administration and the National Institutes of Ileallh. From the Erwymolohr Research Iaboratory, Martinez Veterans Admini.tra- tion Medical Center, Martinez, Cal.; and Ihe DepartnseM of Internal Medi- eine, llnivenNy of ('aliforni. Schod of Medicine, Davs. 1 31

METABOLIC ACTIVITY OF nFVEI.OPINO RAT LUNQ I ong slicet prepared from rats aged 2•12 days were e.amincd for in- corpo.atinn of Ihe labeled pnxunors, 'H•palmibte, "Cthnlirre, "('-proline and '11 thscosamine, into lipid, protein and tlycoprotein. Results of this study of developmental chankn in tst hrn6 composition and biosynthetc capacity show that the immediate perinalal pcriod is one of very high mclabc tic activity and that the synthesis of structural components precedes thal o(' secrelory products. Specillcstly, protein conleal e.prnsed as m6/ 100 rn6 ~ret weight inereased from 1.95 -t 0 25 in fetal hrnp lo 3.0 r 0 16 at birth, aml remained alnw,t unchanged during the Arst two weeks after birth. Individurl measure- menls ahr.wed that palmi/a1e incorporaliow increased (rom 18.2 t 21' nrnd/m6 p.ntein/h two days before birth to 40 0 t 1.0 nmol/mg protein/h three days after birth, and decreased to 24 0 t 2.3 wmol/mg prolein/h at 12 d rys of age. At all akn /e+red, mwe than 75% of the pshnitic acid taken up ty /he lun6 was In esreriAed form. Incorporation of psimilie acid into lecithin was hi6h- est one day after birth. Clroline Moorpor.tbn ksto keithin larcraned ei6ht- fold between two days before and ewe day after birth and decreased 60% by 12 days of age. There was no dHlererroe between the Lcorporation rate at 12 days and that in the .du11 hrng. Total hn.it pbospholipid increased 220% between two days before birth and the day of birth, and ch:N+ged little thereafter. Phoaphalidyl choline rnade up 30-607i of hanol phosplolipids at all ages Proline irrcorporation iMo hrnlf protein was highest In Ih- perinatal period, while glucassmine incorporaliow was highest in the fetal hrag, but decreascd 67% one day after birth. H.nrorA, AI. N J. elotop of rA, kronrc )9:117-126, 1f111. Other aayr.rf r National Inalilutes of lleabh. From the Departmenls o1 rediatria and PbpsiokM and SiophTsics, Oeor6e- town University Medical School. Washiwli/os, D.C. EFFECT OP DEXAMPTHASONe ON LIPOPROTEIN LIPASE ACT1VfTY OP FETAL RAT LUNO In thir study of the e/lects of a eortkflosterokd oe aryme aaivit7, det- amethasoee ws administered by eowtiauotra subcutatreorr infusions to preR- nuN rats from the 16th day of pestatiort. Adminiatralion of the hormone markedly affected malernal and fetal wei6/M pin, fetal 1wr6:body weight ratio and lipoprotein lip..e activity o( the lurrt Cumulative maternal weight gain lrom days 1 f-21 of 6eslatiqw was 60 t 1,0 p M eontrol and 70 t 10 A in des.rnclMsonctreated rats. Fetal weight at 22 days of Aestatksrr .nd one day .Iter birth was 5.0 t 0.)f and 8.6 t0.)0 in tonlyd and •.65:0.26 .nd 3.1tO.11 in desarnetha+one-/reated rats. Ths ratio of lung weight to body weight was lower throushout the Ias1 Ave days of gestatioa In de.ametha- sone•treated than in eon/rd rats. Deaameth.sone administralion led to a Iwo- to threefold increase in lipoprokin lipase activity levels in fetal ra/ lung at 1• .nd 20 days' foetlstiow and prevented the decline is enryme activity sMrtly he/ewe ba.tt 71re resuMs of this study show that IipopnNein Ipase, rhe rnifn.e rhal rrgul.les the upuke of Iri6lrceridr Iatry acids by earshepatie tissues, also is induced prematurely in the fetal (ung aflcr maternal cortico- steroid administration. The early appearance and constant high activity kvel of this enzyme indicates an adcquate supply of triollyceride fatty acids, and suMests that the increased uptake of these fatty acids by the lung could be a conlribulory factor to corticoueroid-enhanced rurfactaM synthesis. Mostello. D. 1., ILrnorli, U. and Hamaslr. P. eloloq of the Neon.re 10:121-12t, 1921. OtA.r aurprfr National lnatilutes of HeaNh. Frvm the Departments of Pediatrics and Physiololly and Biophysic., (korge- tow. University Medicd School, Washhrgton, D.C. DP.POSITION AND DISt7tllUTION OF THE TOTAL rARTICULATE MAT'1 F.R OF CK3AREM-E SMOKE IN MICE USING A LARaE- CAt'ACITY SMOKE EXPOSURE SYSTEM BC)FI/Cum (CS7 SI/CunM it ClH/Aao( Ctun) ruak aod female tdoa were eaposcd in this study to whole cigarette snwke using a standard anwk- inp re6inrcn with a new automatic smoke espoture wuchine (SEM 11). TM SF.M 11 is a lar6etapacity (dti11 mice) dynamic tanoke exposure system in which snroke is rowed through the animal eorrldrwrrenl syslern as a aoa~ tieuously Aowing aream. Mice are restrained about the neck in stock-liks holders for "noscawl7~ exposure. Using uandard onoke exposure co.di. tiom, the deposition and internal diMriMrlion of Ihe total partieulate t+ratw (TPM) /ran cigarette smote was detenniwed iw Mae rnice. Results sb.ei that: (1) snwke eapo.ure eonditioas cam be varied so that depositiw lroru 30 to 200 rs TPM/kwrg eaw be oMained: (2) 00-i0li o1 the TPM deposi- /iar was found in the respiratorp tissues; (3) the anouse-lo-nrourr variados for TPM deposition in ptilrrroa.ry tissue was - 20Ri; (4) similar depod- tion and distribution of TPM waa ohaerred iw rnak aad /ernale wdoe, a.d (3) deposition and d'ntribwiow of TPM was sw/ altered in mice eaposed Io smoke oe a daily basis over a sia-/noelh period of tintt. Heruy. C.1. et d. (Mkrobielo6kd Araori.rei) Toakolory.nd Aoplied Ph.rnr.roloRr 3/:79l-109, H/1. Frora the Deputmen/ of Experimental Oncolopr and the Deputnrent of t1a chemical Oncolo6y, Microbiolo6ical Assoeiatea, Ikstheada, Md., and Awaly- tical Chemistry Division, Oak Ridge National Laboralory, Oak Ridp„ Tea.. I AIRWAYS RFSPONSF. TO INHALED TOSACCO SMMCLr: TIME COURSE. DOSE DEPENDEN('E AND EFFECT OF VOLUME HISTORY The time course and dose dependence of the brenchocowstrictiw respoae to inhatation of tobseco snwke are rcpurted here. In this study, airways re- siuaoct (Raw). lhoracic gas volume and Ihe nwimd eapirMory Aow vo1- ume curve were measured beforc, a(ter each puA, and up to 30 rniwtes dter the last puff of three Ixands of cigarettes with diAcrcru cornpositio.. Raw increased si6ni/icantly with all three bra.ds of cigarettes after one pull. 32 1 33 .

, 0 w w J The maximum eflccl was reached sller three puffs. Instantaneous flow al 50% of vital capacity decteased si6ai/kanlly wilh cigarettes high in nicotine content, but this did not happen when a low nicotine (0.31 mg) cigarette was smoked. Instantaneous flow at 75% of vild eapaeity out (FFF„) in- ereased signiAcantly 30 minutes after the low nicotine cigarette was snsoked. A deep inspiration prior to Raw delermination reduced by approximalely one third the bronchoconstrictor effect of cigarette smoke. A)l effects were reversible within 30 minutes excxpt Ihe delayed effect of the low nicotine cigarette on the FEF,,. On the basia of this work, it seems th.,t the probable sile of action of tobacco snwka is i. Ihe large and central airways. The bronchoeonstrictor effect rapidfy resdw a plateau. However, a delayed broe- chodi/stion of the smaR airways observed slter the low okotine cigarette was smoked might represent snwher response usually masked by odrer long-acting componeas is smoke. Tsveira Da Silva. A M. snd K.w.e.A, r. Rrrplrrlon 11:96-105, 1961. Frnm the Drp.rtment of Medicine. Physiology and Biophysii,, Georgetown Universi/y School of Medicine. Washington. D. C. RESPIRATORY DEPRPSSION PRODUCED BY ACTIVATION OF OABA R ECF.P rORS IN H INDBR AIN OF CAT The aim of this study was so ehsracleri:e the respiraory effect of r•sminobMrtyric acid (OABA) administered directly into the ( NS. Muscimol, a specific GABA•receptor sgowisl dru& was sho used because it is more potenl and longer Iasling than OABA. For this investigslion, respiratory re- aponses to activation of OABA recepon in the hindhrain were measured in ehlaralose-snesthNi:ed ests rsin6 a Fkisch pneumotachograph. GABA re- eeplors were activated by intracis/ernal injections of muscimol and OABA. Museinwl (0.014.65 pg) administered so seven animsh caused a depression of respiratory activity with apnea occurring in each animal. Before apnes occwred, a decrease In tidal olum» was osacrved. Respiratory rate sad iw- spiratory and expiratory duratiows were unchanged. OABA (A 05-12.1 S mg) adminiskred to /ve anie»b proAuc'ed the same effect as nrncimol on res~ piralory aNivity. Apnes produced by both ye.ts bss reversed by iMra- cislernal administration of the OA1A-receplor antagonist drug, bkucutine. Administration of bitueuRine to four naive animals ixreas.d tidal volume but had sro eRect on either respiratory rate or inspiratory duration. These resuMs indicate that OABA and rrwseimol. which activate GABA recepbrs, cause respiratory depression. it seems slw, on the basis of Ibe data obtained from both agonists and the sntagoniM of OABA recep/ors. that OABA might be an important CNS neurolranaeilter tekulalin6 pulmonary ventilation. Ysmada. K. A.. N.wesli. r., and Oillis, R. A. lournl of Applied PAysiolon: Respirat. Environ. Exercise Physiol. 31(S):- 127a-1286, 1981. O/b.r supportr American Heart Association. t'rnm the MpartmeMS of Pharmacology and Physiology. fieorgelown Uni- vcrsuy Schcwds of Mcdscrnc and [kntistry. Washington. 1) C. 34 t OPTIMAL ('ONqIT1ONS FOR CELLFREE SYNTHESIS OF FLASTIN In this biochemical study of elaslin syn/hesis, particular emphasis was placed on the development of opimal conditions for the translation of elastin mRNA in the mRNA-dcpendenl rabbit reliculocyle lysate syslem. Using taal RNA isolated from embryonic chick sonae as the source of exokenous RNA. the following determinations were made- 1. The effect of heating the RNA to different temperatures just prior to IranslMion, 2. the prest.ce in the translation assay of bolh proteins that resrdt from the translation of elaslin mRNA, and 3. the effect of different eonctnlrstiorn of emilnesium acqate, potassium chloride, spermidine, crealine phosphale. ATP, and OTIi' on the Iraoslalion of elastia mRNA as seen by immurqpreeipilstion. Results showed that heating of the RNA prior to translation signiAcantly enhanced total protein synthesis, elaslin synthesis, and the symhesis of pruwcins possessing molecular weights of greater than 90,000. The development of this Ira.s- lation syslem, which is optinal lor elastin syMhesis, should prove to be iw- valuabk for studying the mechanisms of control of elastin biosynlhesis. Karr, S. R., Rich, C. B., Fosrrr, l. A. and Pr:ybyls, A. Coll.grn RtirrrA 1911. Otbrr support: Natioaal Institutes of Ileahh and the National Fousdatie.- March of Dimes. From the Departmenl of Biochemistry. University of Georgia. Athens. IMPROVED METIIODOLOOIES FOR THE ISOLATION AND PURIFICATION OF TROPOELASTIN Strvcturd anA tnrtabolic studies of elasd" have been hampered I. as psst by the difficulties involved in isolating wr/Reient quantities of tropoelarls from lathyrilic anim.l s~sakls. To otwntttw this problem, several tweiods have been proposed to increase the ridd& of puried tropoelastia 1. M@ study reported here, tday-ald chicks were used for the i.ductioo of Ialhy- rism in order to take advantage of Ihe teporled Sursl of elaslis sHrthaia during the second and third weeks of chick development. In sdditiow /o dr lathyro6ett that was added then to the diet of the chicks, r•aminocaproic acid was added. The increased age of chicks prior So Isthyroken adminislralios plus the addition of the trypsin-like ensyrlte inhibilor, r-aminocaproic scid. to the diet resulted in a 110% increase in the yield of Iropoelasli.. This method for induction of lathyrism in chicks o/lers the advantyes of ho/h increased yieWs of Iropoelasliw and the convenience of dealing with smsBer 6rory+s of chicks. In addition, a ealiowtxchan6e system employing a CM- eellulo.e column was devchrped in ,his s/udy, which allows complcle removal of minor acidic protein contaminantA Irom tropoelastin. Foner, l. A. er .f. Anefrrkd eiorhe.nl.rry 10a:2)3-2 )6, 1980. OOther support: National Inslitules of Health. From the Ikpartnscnl of Biochenmi.lry. University of (kor6ia. Athens. 3S

MF.AS(IRtMFNT OF I:I.ASTIN DFGRADA I ION IN VIVO BY DISM//SINE RAUIUIMMUNOASSAY Tksnxsine is a cross link mino acid found only in clashn, .nJ urinary desrno.ine e.cretion is elevated as a result of increased elashn degradation in the hody. A chemical mcthod for rneasuremenl of urinary desmosine has been utfired helwe. fwt the present paper describes a highly sensitive radio- immunoaatay for desmosine which is capable of detecting as little as 0.2 oR n/ this conspound in acid hydrdysales of urine. the nsethod does not delecl other amino acrds, incladinp Iysine (the precursor of the desnwnine cross- links), hu1 rather is specific for desmosine. Preliminary data on desnsosine eacretion in normal tsonsnaken ad is a tnnaM drvup of heavy smokers with a variety of pulmonary disordcrs are presented here. It seems, so 1ar, that the method dr.cussed here may prove useful in monitoring elastin break- down m several patho/opscsl watn, iacludirrd pulmonary emphysema. Ilarel, S. Yu, S. Y., LnoO, A.. Ilurewits, A., and Bergas/sky. E. 11. RrJlrne furopr..w Jr 1'Irlriop.nhulurir R.rpi.woirr I6(wpp1./:7t.N1. 1990. Othrr aarpport: U. S. National Ileart, Lund and B/ood Institute. Fron+ the Ihp+rtmems of Pathology and Medicine. State Univcrsity of New York al Stony Brook, Stony Brook; and the Veterans Administration Ilos- pstsl, St. I ouis. PRF.VFNTION OF F 1. ASTASF -INDUCED FXPERIMENTAI. I:.MPIIYSEMA BY A CYNiltt /IC ELASTASt INIIIBI IOR ADMINISTERFFI) ORALLY Since there have recently been several reports of prevention of esperi- mentally-induceJ emphysema by Irea/menl with praNeinaK inhibitors, an al- templ was made here tu test the eRcctivenes of oral aJministratwrn of nrelh- oayurccinyl slanyl alanyl prolyl-valine chloromelhyl kelone, a pacnt inhibitor of clastase. in an animal model of emphysema. Results of the pre.enl eaper- imenls showed that, following adminislrNion of the inhibitor ta mice by slomxh tuhe, a portion of the labelled ehWromelAyl ketone cou/J be recov- ered in an active form in hmg wash. Signifkantly, animals given a single oral dose of lll0 pg of the synthetic elaslasoinhibilur, 13 minutes prior lo introduction of si units of porcine pancreatic elaslase into the hrngs, were corrspktely protected against alveolar deformation (as judged fr.Mn mean linear intercept values four weeks later). Treatmenl with the inhititor al tima earlier Ihan 13 mrmrles Ir•fore enzyme chalknge or from IS minutes to •N hours afler enzyme challenge did no1 protect. These results provide direct evidence that chloromethyl ketones can he effective when given orally and. thu% wrth further sNdy. they may snme day pr»ve prsctical as pHCnlra prophy- lactic agents in populations at risk for development of obstructive pulmonary disease. lasn0. A. and 1learing. R. /rul/rnn 1 urorrwl Jr rhriioparAolorir Rrrpl...uarr 16(suppl / 199 l/11, 19NQ. Oflrer aupporl: U. S. Public Heahh Service. From the Ikparlmeol of Pathology, Health Sciences Center, Sratc Urversily of New York sl Stony Brook, Stony Brook. POTENTIAL MEDIATOR OF INFLAMMATION: PHAaOCYTE- DERIVED OXIDANTS SUPPRESS 7HE E1ASTASE-INHIfalTORY CAPA('l1Y OF ALPHA.-PROTEINASE INHIBITOR IN VITRO Within their cytoplssmic tranuks, hunsM kukocylea contaie protessam that are capable of degrading connective lisws slrsrchues. Iw the arudy pra. setwed here, i1 can be wen that human polymorphorNSckar (PMN) knkocytea, nsonocytes (MN(') and pdrnonary alveolar macroph.ttes (PAM) that wetr nin•.dated in rUro by plwsrbol myristate acqate (PMA) released reactivc osygen specin that sre ahle to suppress the elastne inhibilorr capacity (E1C) of human serum. Immunocleclrophoresik wirsR nmtsodies apinsl n/.prs teinase inhrhi/or (.,-Pi) and elaslase showed MN i.aelivalion of .,-Ps was responsible lor the decreased serum EIC. TreYrneM of plsaRocyle-insetivaled serum with a reducing agent (dilhiolhreilol) resuNed in siRnifkan/ reco.ery of EK'. suggesting that w,-Pi had been oaidslirely, inactivated. Setww f21C was partially protected by superoside diunwase or eaNalsse. and hydsoges peroaife alone had no effect on serum E/C. Tlws. neither /IsO, nor Or abne, but a producl of the two, may have osidalively inaclivated .,-PI. !o suppon of this observalion, it was noted thN MNC and PMN from a pa- ienl with chronic tranulomatosis d'tsease, in which getseralion of O, .e/ 11.0, is depresxd, failed to inactivate w,-Pi when e.posed to PMA. On the basis of this and several other esperimerud noliwp, it appears that osidalive i.ac- Iivalion of w,-Pi in the microenvironmeM of infaamnsaory eells, at siles of scwe or chronic adlammslion. may allow proleases released Irorn lhese eeRs to damage adjacent connective lissue eonsponeeb reore readily. I Csrp, H. and /awoe. A. lorrnaf o/ Clink.f fnrrsriNrrion 66:fR7-!!S, 1980. O/Aer nrpperf: U. S. Public Health Service. From the Department of Pathology. Heahh Sciences Center, State University of New York at Swny Brook, Stony Mook. URINARY EXCRETION OF DESMOSINE IN PATIENTS WtTH SEVERli BURNS Burns involving either partial or /oR-Ihiekness of skiw iwvariably dsl.- age connective tissue layers, which largely consist of elaatiw snd eoNyeo fibers. In this auempt in study the degradation of elas/in in burned pMie.ts, urinary escretion of total dssmosine was measwed'by a r.dioimrnuwo.asr in 10 severely burned adult maks, as well as in 1• normd adult rnab ooms- trols. Total urinary desmosine was sidni/kanlly ekvNed in all the s.apla in the burned patrents, who had injuries involving more Ih.w N!L of total body wrface area lhe values of 21•hcwr urinary desmosine for the patients 36 1 37