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matic hy.lrocarhnns lhus. hefnre slarlinR any large scate in virrn Irsmforma- non prrrFram. p~ucntial human ccll lines should he screened su a, tu idcnti(y Ihone with high entynx Icvclt Thn approach has been parliaulerly u.eful in eluci&linR the rclatinn.hips between hydrocarbon metaholumR entymc activity and sensdivity hr hydrocarhon induced cytoloxicNy Ilere. the authurs cumpare two prraedures lor measuring the enzymes lhe one desiRned tot mcasure henzol a1anthracene ornhrced aryl hydrocarbon hydroxyla.e activity could Jetecl and quants(y enzymc aclivitrcs in low passaRe rodent cells. hut could not rcproducihly detc.t Icvelt in intermediate or high paaaRe nkrute, rat. or human cclls. The method dcsiRned to measure the ahilily of a cell to convcrt henrolal- pyrene Irom an orRamc-u>tuhk to an aqueous acetune-soluhle (ornr. proved more reproducihle When nwdiM1ed, this particular technique proved to he an e(fective screening tru for the detection of those lines with higher levels of hydrocarbon metal.M.hzing enzymn. Kouri, R F, Kiefer, R and Zimmerman, F. M(MirroAlototirol .+lrrnriorrr) In VJrro IO(1 i 21 :1t1-2S, 1974 OtArr .nPport: National Imtilules of Heallh. From the nepartmenl of Virall'hemieal Oncology, Microbiological /lssociates, Bethesda, Md GFNf•.TIC CONTROI. OF Sl1SCFPTIBILITY TO )-MFTFIYI.- CIIOLANTIIRI:Nh INf)11CF1) Sl1BCl1TANEOl1S SARCOMAS The aulhors report on their use of a mouse genetic system to estend their observations on the rclationship between aryl hydrocarl.nn hydrosylau ( AHFI ) inducil+shty and sensitivity to 1 methylcholan+htene ( M('A ) induced tumorrtenesis Ohurvatrnm reveal thal AFllt inducible mice are pproximately 12 times more sensitive to MCA induced (umorrRencsis than their n.roinJucihk Idtermatcs live type of parenlal cross (maternal influence) plays n.r role in this sensitivity IMtt regulated espression of the Rroup-specific (R+) anuten of type-(' RNA virusc%, although also sepcgating in this genetic systcm, d(oes not seem to play a major role in this enhanced susceptibility to M('A careino- Rennis Results are discussed with the view that the enhanced sen.nivily of the AIIIt inducrhk arumals probably results from rapid and e/licirst metab- olism of the chemical to its ultimate carcinogenic form. Recent information suRResls that these ohservatiom may he analoRous to what occuirs in human Ix.pulalions wherein the degree of AF111 indueilrihl) is genetically cnntrolled (probably by a single Intns), and inheritance seems to he crxlominiint Thus, there may he a correlation hetween the presence of (or susceptibility to) lung careinomas and the alkks regulating eapression of high or inlermrdule in- ducihdrly II so. the genetic system presented here may provide a vrluabk model for studying this probkm in human populaliona Krwri. R 1: , Ra/rre. If , 111, and Whitmire, C. FWurnMolurirrnl Ai.n.iotrr) Inrr.nononar lournar of Cancer I)( S ): 714 720, 1974 OIher .upport : Natu+nal Inslautes of Ilcalth 1'rom the tkparv.Knt of Vual ( hemKal Oncology. Micruhrusu`K al A.%Ki,.tes, Iklhcs+l.. Md F.N('AI'SlI1 A1ION OF I.YMI'll(X'YlE 1)NA IIY VI:SI('l11 AR SIOMAIIIIS VIRUS Several aninul viru.cs are known In incnrporatc hmt DNA during their replication and maluralioo, hut this procest was Rcner.lly thought to be char- aclcrutic o( uncuRemc DNA viruses with a nuclear phase of rcpltcalNrn. Ilere, howcvcr, the awhors denonstrate that an RNA vinus which is not considered oncogcnic, has no known nuckar phase in its replKation, has its own RNA dependent RNA polynserase, and .huts ufl host DNA synthesis can contain tip tu 10% of its genetic mass as I)NA. In addition to vrral RNA, vesKubr sUxnatitis virus ( VSV ), grown on the WILz-)A line of cunlinuously growing human lymphocytes, contains DNA which is found in twth the B and 1 virus particks and is resntanl lo deosyribunuck.se Ihrt I)NA is intimately asso- culed with the virus and appears to he incorporated into the viral rihonucko- protcin slruclure produced by treating VSV with Nonidet P40 followed by ('s('1 isopycnic banding. In con(rast, virus grown on B11K 21 ('1 ) flbroblas(s has no detcclabk DNA. The virus-associaled 1)NA has n isopycnic denrty of I 699 if X cm a in Cs('I, identical to that of human hNA Its average molecular weight is 90 s 1(P, as delermined by velocity sedimentation in sucrose density Rradienls, and studies of its conlour knglh in the ekclron microscope. The DNA'DNA reastoeialion kinHres of this particular nucleic acid demonstrate that it is of host origm and rule out the possihi6ty that u originates from contaminating microorganisms or minrchondria T he analytical cnmpksuy of Ihe virus assoeialed DNA shows that S(1Rr of its sequences are homologous to the repeated DNA sequences o( human DNA and that the remaining 50% are homologous to human unique scquenccs On Ihe avcrage, there is one mokcuk of DNA for each (our virion puticles Whether or not viruses such as VSV are capable of oncoteme inlornulK)n "transducuon" ro unknown, but it is a possibility which can he tested. Kin6sbury, I). T and Lernrr, R. A. Procrrdingr of the Nnriann( .IraJcnry of Scienrrr of the (In/Urd Srarrs of Amrrica, 71 (S):175)-I7S7, 1974. Otlrir support: National Cancer Inslitute, National Fuundation .- March of I7imes, and U. S Public Ilealth Service. From the I)cpartment of Medical Microbioloty, llnivctnty of C.lifurnu ('01 kgc of Mcdicine. Irvine, and the (kpartment of Fsperimental Pathobgy, Scripps ('Iinic and Research ('oundauun, 1 a)olla. ('.1 I)11lFFRFNIIAI RI S1'()NS1' (1F SN111'S .1N1) ('17 Ifl A('K MI(':= T(1 ('lIRONI(' INIIAI AII(1N (IF ('IGAR1 1"11 SMOKI' PIll MONARY ('AR('IN(KiFNUSIS ANI) VAS('l/l AR AI II RA11(/NS IN I l1N(i ANt) tI1 AR I 114% gcrirtiC Jd(crcn(ct helwecn nuume shjrf,. infli1(ncc Iheu rrstx.n.e /u chronic inh.lrtion of cigarette snwke? Ihe qur+tuwn ts e.pl.ured in IMs study 17 16

comparing the effccts of chronic smoke inhalation in Iwo slrains of mice. Snell's and ('57 Black, sekctcd for their respcctivc tendencies wuh aging to develop sponuneous IunR cancen and vascular alreralwms in lung and heart plus lymphoid cell infilrratrons arnund bronchioks and bronchi Only rarcly do Sncll's mice spuntaneomly develop nrch v.scular ksitxrs. whilt (-S7 Hlack mice do not develr,p spnntaneous lung cancers The latter strain rs alsro espc- cially well known for its escelknt immune response. lhe two nsouse types indeed had a notably different response to chronic smoke inhalition After inhaling whole cigarette smoke •nd especially its gas vapor phase. Snell's mice developed lun6 sdemxarcinomu mote frequently and earlier than the cnntrol animaH, while the ('S7 Btacks did not. llwae mice, however. had a higher incidence of vascular change in lung and heart after w-holr cigarette smoke inhalation - and nnt after that of the ps vapor phase - while no vrch effect was noted in the Snell's strain. Resuhs suqes( lhat cigarette smnke enhances esisting genetic susceptrhililies in these Iwo strains, Mr( immunological pro- cesses may play a role in these differential responses as well Chronic c.(+rnure to the las vapor phase decreased the total number of immune competent cells per spleen in Snell's mice and slill more prolonged esposure to higher snsoking kvcls Ir,wertd their antibody hinding capacity to polyvinyl pyrn.lidnne. -Ihese observations wtkcu that the presence of lymphoid cell inflllratrons in C57 Black mice may alter their lunr: response to cigarette smoke. 1 he relation- ship between lymphocytes and the frequency of sponlaneous tumun in ddler- ent strarns of animals and their responsn 1o cigarette smoke merits further invesligalion. - I.ru(hrrnhrrrrr, (' and I ttxhtcnherger, R. Onrnloty 29: 122 119. 1974 I7rom the Swiss Institutc for I'spcrimental Cancer Rcseuch, (.ausanne, Swittcr- land. TNE FXPFRIMFNTAI. FXPI ORAl1ON OF EIEALTIi DAMAGING F'A('TORS IN ('IGARE:TIE SMOKE AlthougA several skklies have wtgested that cigarette smoking is a health hazard, being particularly important in the etiology ol human lunfi canccr, it is pteuntly stdl ddlicult to determine which spcufic snN,ke components re nrnrous The main reasons for this uncertainty are the largc numf.er o( cmnke components with yet unkm)wn hit,IrKical eRects and the drflrculty ol duphcal- ing the human snwrkrng hahil in lafxxatory animds Accor,hnR Irr rh: :urrhors. htrwever, the available e.pcnmental data juslify the h,lh,wrnp cn.r.luswros (1) Ilealth damaging factors, such as Ihose contrnhulrnR Ir, carun„Rcnesrs. are found in both particulate and gas vapor phascs of cigarette snw,ke (2) Painting erperrmcnts in animals and esposure o/ cultures tn the p:urrculate phase, namely ciR.urrlt "tar," have shown that Ihis (rxlr.n hn .a.trm,/,~tnic pra1rrtrrs in skrn trachra and laryn (1) Inhal.hon CRprlnnrnll ur anun.ls and r.l.,snrc nl nnn.l anJ hnman IunR cultures rn frc.h crRarcnr •r -kc have dcrnnmlr.lcd tnh. c n. nl ul t art,nr,Rcnest. nul unly aller c%lw,.urr 1„ whrde ciRarelte smnke but also afler esposure 10 the gas vapor phase alone (4) 11 is urgent to characterize the reaponarhk components in particulate and gas vapor phase and their mechanism of action. (S) The public should he informed Ihal, based on present knowkdRe, ci6are11es with either reduced "lar" or reduced gas vapor comtiluents cannot be consideved sa(e, that is, smoking Iheae don nol eliminate damage to health such as the risk of lung cancer. Leucbtrnbrrtrr. C. and Leuchtenber`er, R. Sotlal- und lydlvrnrlvmrditln 19:41-43, 1971. Ottier support: A. S. F. C., Switzerland. From the Department of Eaperimenld ('ylochemislry, Swiss Inalatute for F~t- perimenlal Cancer Research. Lausanne, Switrerland. MODIFIERS OF CARCIN('iENFSIS The environmenl undoubtedly delermines to some e.tent the oulcome o( Interaction between chemical carcinogens and cell conslilutnls. Any envfron- menlal factor affecting Ihis reaction, /herefore, is a modifier of the carcinogenic process, as are sonre of the inherent properties of both carcinogens and cell constituents. While modifiers operate as of the mornenl of contact between cell and carcinoken, Ihey may in facl have been active in conditioning one or the other prior to contact and c.rcinotenesis. Equally important is the possi- bilily that modifiers do not affect the carcinogenic process rtself, hut the devel- opment and growth of the resulting neopl.sm. Consequently, numerous (acton such as cocarcino{ens, promoters and inhibilora of carcinoeenesis, immuno- logical inhibitors of lumor growlh, vehicks and roules of administration, and host Lclors can be considered nwddiers. The aulhor's objective in this knglhy review dealing with chemical modifiers is to make those working toward the development of an effective system of cancer prevention thoroughly aware of the available data. I1is comprehensive discussion presents the various aspccu of the subject under five main headrngs: ( I I Importarke of ( hemical ('arcino- gens for Man; (2) Biological Modifkrs; (3) Physical Modifsers of ('arcinu genesis; (4) Chemical Modrfkn; and (5) niscussuon of Inhrhuors of ('hemrcal ('arcinotencsis Graphs and tahles are used whcrcver necessary arxl a very long bibliography is appended 1fomAurtrr. F In: IlornhurRer, F. (ed ): The rlrplrrpatAnloty r,/ ('uncn Bruluay an.! S/o- cArmiirry, Basel: S. Karger, 1971, vol 1, pp I10-1Stt Other .upp+.rtf II S Pub6c Health Servite, Amencan ('arxrr Sacrtty, s'annie FF kippcl 1'uurulatrnn. Viusrma and 1) K I rklw,g 1 uunJatrtws, and Bio-Rcscuch ('ornullants, Inc. From the Bio Rcscarch Institutc. ('ambndte. Mass 18 19

i r a, m m 11. The Rr.piratory System INTRAPl11.MONARY NFl1RO.EPITHELIAI. BODIFS: 1IYPOXIA-SF.NSITIVE NEURO(CHQMO-)RECEPTORS It has long been postulated that in addilion to central and peripheral chemorecepton. the lung contains inlrapukrwnary air chemoreceptors which have a regulatory function governed by acrodonin; but in spite of sup~ortinR physiologic evidence, their presence hai wol boew histologically demon-.trated. Investigating this problem. these aulhors have previously identrfkd and de- scribed the socalled neuroepilhelid bodies (Nee's) in the mammalian lung, including that of man, and have shown that they contain serotonin as well as other substances. E(ere they report on their rrludy, by light and ekclron micro- scopy, of NEB's in rahhits subjected so esper(menlal hypoaia tJrder these con- dilions. NEB's were found to secrete the dense-cored, serolonin containing vesicks at their basal vascular pok. The aulhors propose that among their various possible neurorecep/or functiom, lhese NE<!'s form an intrapulmonary chemoreorplor system which is sensitive to hyposi and supplementary to such well known central and peripheral chanoreceplon as the carotid body. They secrete not only aerolonin 1" probably also associated amines or peptides which could inAuence the pulmonary vasoconstriclor response, and are regu- latcd by the central nervous syslem. Lsrwrryru, 1 M. and Cokelaere, M. Eaptrienti. 29(11) • 1)d4-I )s6, 1973. Othar aurport: Nationard Foods voor WNenschappcCiik Onderzock (Bel- gium). From the I,aboratory of Ilistopathoiogy. Katholicke l)niveraiteit te I euven School of Medicine. l.euven, Belgium. NYPOXIA-SENSfTIVE NEURO F.PITHEI.IAI. BOE)IFS: IrITRAPUI.MONARY SIi('RF:TORY NEURORECEPIORS MODULATED BY 71iE CNS Several routine and silver slaining methods. Fakk's fluorescent amine and other histochemical tcchniques, as well as electron microscopy. wrrc used in an allempl to further elucidate the slruclure and function of the reccntly idcn• lifled mlrapulmonary neuruepithelial bodies (NEB's). lhe lungs obtaincJ from 94 nconatal rabbits and sia neonatal mice showed that: ( I) during hypwra the corpuscular cells of the NIiB's secrete their derne r ioo ni reatment vesicks 111('V's) at their basal vascular poie; (2) E r rescrP Prc the NFB's of odherwise rwvmd animals suRer from a distinct amine dcplctiun. the corpuuular celh displaymg dccreascd yellow fluorescence and emptied or otherwise abnnrmal IX'V's; (I) as observed by ekctron mrcro.copy of acrn.t aections, the NI H's are innervatcd by numerous unrnyelhnaled fiher+ with IKdh allcrcnt hke and cflcrenl hkc endingi which are in synaptic contact auh each 20 orher as wdl as wilh the curpu.cuLr celh; (4) cytochemrcally, corpuscular cells arc posuive for acelykholineslerase; they olso have a positive reaction to Ihe u ~IYcerophosphale dchydrollenase technique and to Sokrr's lead hema- losylm slain for polypeptide- and amine-prrrlucing endocrine cells. lhe au- thon propose, Iherefore. that the NEB's provide an inuapulmonary, hypoua- sensitive ncuro(chemo-)receptor system in addition to the well<stablished un- tral and peripheral (r;., carotid body) chemorccepton. TFey conuin and secrete serotonin, and probably related amines or peplides, which could ir.tlu- ence the pulmonary vasoconslriclor response. Also, according to classic nwr- phobgic crileria, the NEB'a possess both aRerenl and eflerenl innervation. Their various other possible functions in normal and diseased lungs are mes- lioocd briefly. LAuwrryru, !. M. arnd Cokclaere, M. Zclrschrl/t /ur Zcfl/orulirnt rnJ m/RrorAoPucAe Anaromie 14S(4):121-540, 197). Other support: Nalionaal Fonda voor Wetenschappeliik Onder7oek (BeL lium). From the IAboralory of Hislopalhology, Katholicke Universiteil it l.cuvc. School of Medicine, Leuven, Belgium. BRONCNIOLAR NEIJRO-EPITIIP.LIAL BODIES IN THE NEONATAL MOUSE LUNGS The neuroepithelial bodies in mammalian airways are inlramucoul oor- puscks composed of nonciliated cylindrical epilhelial cells. lhese contain ckre- ly-packed oval nucki with compicuous peripheral chromatin condensalions and rather dark granular cyloplasm; they estend from the basement membrane to the airway lumen. 1111rasaruclurally, these speciaheed cells are associated with inlraepilhdiat aaons which k+se their Schwann cell shealhs. This makes them difficull to identify and has led some investigators to consider them cell processes ralher Ihan nerves. The present paper aims to clarify the ultrastruc- lural characlerislics of these uons in newborn mice by presenting results of both light and ekctron microscopy studies which demonstrate the orillin of intraepithelial aaom. Electron microscopy of lung lissue from a total of 120 speciPic-palhogen (ree Swir mice, ('D-1 slrain, one to nine days old, showed that the nonmyclinated ason under the neurocpinhelial hMdy penctrates the baul lamina an) emcrs the epitheha( (ayrr After pcnetratK.n, the intracpuhclral ason conlaining numerous mNochondria k»es its Schwann cell shealh. Lcuxnes enlarged nd rami(ies anang the eprthelul cells. 1 he esrahlishnscnl of mito chondria-rich structures as a.ons is particularly significant because similar slruclures associated with the granular lKulrachruky) cells have rccemly been idenliHed as cell processes and nw ncrvcs E'urthermnre, the aulhnrs ruqrsl, their rnrxphnQiRi..l nrganitalwrn supprvts Ihe prnbahilrly rhat inuacpidvchal nerves together with their asxxialed cells function as scnsury rrccptors in the bronchioks. 21

Flung, K Snd Loosli, C G The Amirican lournol of Anaronry 140(2):191-200, 1974. nrfs.r support: Fnvironmcntal Protection Agency. Howard Huahes Fmploy- ccs Give Once ('lub, and the Hastings Foundation Fund of the University of Southern California. From the l')epartnscnts of Analomy. Pathology, and Mcdicine. University of Southern Cahlorma School of Medicine, Lod Angeks. T1tF. ROl-E: OF TIIF PUl MONARY LYMPIIATICS IN THIi [)hFFNSIiS OF THE 1)ISTA1. l.l1N(7t MORPIInIlXilCAI. ANI) FXPFRIMENTAL SIUDIFS OP TNE TRANSPORT ME?CHANISMS OF INTRATRACHEALLY INSTILLATFI) PARTICLIS This paper, which also contains a lxief review of the atahori earlier studies on the microscopic anatomy and ullraslructure of the pulmrsnary lym- phatics, presents an espenmenul study of the uptake and renwval of inlra- Irachcally instilled carbon and ferritin parlicks from the alveoli of newborn rabbits Results show that: (1) in the alveolar lumen, (errilin particles mis much rrxxe easdy with the surfactanl material than carbon parlicles, although both are pha6ocytrred by alveolar macrophages and neutrophds: (2) since the alveolar cells are tightly inierconnected, formrng an impermcahle harrier. the tracer parsicks can reach the imrr.utral tissue cnmparlment only by a Irans- cellular route lypc I alveolar cells seem to have a vcsicular type of transport for fcrrrtdn and carhon, with fairly large nurnhers of Icrrrun mulcculc% hcrng phagocytized and apparently digc.icd in sccundary lysosomes, type It alveolar ccllb also phaprcylrzc both Iraccrs, npecully lcrntrn, strongly suKgctuns a double function for this cell, namely sccrelwtn (rI , surfactantl and phaRr><y- tosis (c t., ferntm, carlxtn and lamellar material); (1) inlcrstrtul connective tissue cells of debatable origin also contain phagocytized carbon or lerrnin particks; (4) the tracers very rapidly appear in the lymphatic lumina mainly through the open inlercellular lymphatic junctions which act as inkl valves activated by the anchoring fllaments. Lymphatic endolhelial cells also phagocy- tize ferrilin particks in large vacuoks and secondary lysosomes: ferritin par- ticlcs .re seen in micropinocytolic vesicles and coated vcsicks; (S) blood capillaries in the inter-alveolar sepra are morphologically much less adapted for the uptake and removal of particks than the lymphatics and seem lo play a minor rtdt in ckarancc• displaying only a limited reaction to frrrrtin hul no1 to cubnn; (6) many parlrcks art retained within the larger nrways to 1re rc• moved by ciliary aclivily; (7) the bronchiolar nonciliatcd cylmdrrcal, or so• called ('lare cclls, are also capabk of pharocytizing (crrrtm molecules. m tn m Laawcryru. 1. hI and Bacrt. 1. H. Ann.b n/ rhr Nrs. Yortt Aradcn+y o/ Stkncrr 221 :2J4 275, 1974 m m D Frnm the I aMratnry of Itrstopathology, KatholieTc Univcnded tc l.euvcn S.h<xrl r-f MrJu inc, I cuvcn, Bclgium. p r- 22 MI:CHANISM OF AC11ON OF a-t-AN111RYPSIN Biochcmiin~ aincal evidcnce su«esla ttiat a-l-antilryp~sin has two inhibitor sites, one cont o posituvely charged residue and the other an aromatic or kucine resirLrc, with both sites having a binding site lur the aarve moiely of serirse proteases. Fsperimenls designed to lesl this hypothesis show that: (t) Irypsin and chymotrypain compete for inhibitory sites on purified a I-an1i- trypsin, suggesting that the inhibitor has the same or overlapping inhibitory sites for Ihese Iwo enzymes; (2) Irypain inactivated by duu.propylphcssplwro- fluoridate fails lo occupy inhibilingsites on a-1 •antitzypsin; (1) a 1-antilrypsin inhibits suhtihsin, a proteolytic enzyme that contams scrine. histidrne, and as- parlic acid residues at its active site in common with mamnulian serrnc pro- teases; (4) a-l-antitrypsin fails to inactivate acetykholincsteuse, a nonpro- Ieolylic enzyme wlwne active site conlains a reactive scrine reudue• sulitestin` that this residue alone is not suflicicnl for rnhibilisn hy s l-antrrrypsin; 1S) treatment uf a-I-anlitrypsin with phcnylRlyosal hydrate blocks its action on trypsin 1.ut not on clsynastrypsin, a change in activity which is presumptive evidence that arsinine residues were modified by the t.catmcnt: antilrypric activity can be regenerated with removal of the blocking Rroups lhex resuhr are consistenl with the hypdhesis that Irypsin and chymutrypsm arc inhibited at two different aites on o-1•anlitrypsin and suggest that the uypsun inhibitory site contains a posiliv6y charged amino acid. Cohcn. A. d. The lournd o/ 8iolock.f Chcmivey 24S(20) :7055-70t9, 1977 Other aupporf r U. S. Public Neallh Service. From the Medical Service. San Francisco General Ilo.prul. and the Specialized ('enter of Research and Department of Medicine. Umvcrsrty of California, San Francisco. Pl1RIFICATInN OF PIIFNOTYPICALLY UNAI.TFRFI) a-I-ANTl l-RYPSIN This procedural paper describes several nsodifkatiom of the method of Shamash nd Rimon that enable ptnification of phenolypically unchanged a l- anlitrypsm from subjccls with phemttypt Pi,,,, lu this improved mahod. thrct Icchniyucs rc employed seetuenlrally protein prccipitariun at conlrulkd p/1 and akrrtw,l arncentratsns, nNlo e.changc adumn chromatography, ar.d prc• parativc polyacrylamide ctl ekcuophoresis All Ihe mrwlrfkations descnAcd hcrern nccur in the third technique T hn merhrKl results in recovery of an average of 12 1% of the Iryptrn inhibiting capacity of the scrunm Ihc a I- anlitrypsin prepared in Ihrs manner mainuins the hercroRcncity in acid starch gel immurnwro-.rcd tel ckclrophorrsrs that characlerues a I anruuypsin in whole scrurn lhn ohstrvarion pn,ves that the pherwuyprc chuaclcnures arc not dependent upon scrum Lrcturs ARo, this purified inhibitor resemhlcs a I- antitrypsin in scrum more closcly than a-l-antrtrypsin prepared by previoualy '1

dcacrnlxd methods In addition- the material h.rs RS-I(X)76 MoloRic activity, misraaes as a single hand in polyacrylamde gc1 clcctrophnresn with sodium dodecylsulhtc. srn1 m/Rrates as  hand with a cathrrl/c shouldcr at ptt 7 S in pnlyacrylamrdc gcl clcclrophoresls ('o/ien. A. e arul Fslt.t, R. Biochimir. cr diephyrira Arra )J6:J99-402, 1971. OIAer sr.pport: Il S Pulslic HeaNh Servioe From the Medical Service. San Fnaeinco General Ilospital; the Department of Medicine. Umveruty of Califorwia School of Medicine. San I:rancisco; and the Chest Division. Pacifk Medical Center. San Francisco. PURIFI('ATION OF TWO POPULATIONS OF IIUMAN ALVFOI.AR MA('ROPIIAGFS FROM SURGICAI. SPECIMENS T?lis paper presents two new methods that have been developed to purify human alveolar macrophatcs relrieved by sransbronchial lavasc lrcxo ulr6ical lung speclmens the MI method yicWs a large number of normal rnacrophaRes that are suruhlc for metabolic and functional studies soon after retricval The cells can he u.ed in suspension cullures or while adherent to tlass T he ucond method partialty purlfics another population of macrophates This method, which involves centnfu6lng the lavatcd fluid through a dense mcdlum, resulU in a 10 fold IxrnhcatK.n of foamy macrophaffes As measured on a surface balance. estracts from these foamy macrophages lower surface Icmlon in a manner similar to that of lung wrfacrsnt Although the origin of this particular maerophalle is rol certaln, a physiolopc role in the metabolism of lung sur- laelanl is sugrsted. CoJ1en. A. e. and Gecry. D. Ansnkan Rtvirw of Rrtpirarory Diitue I0R:972-975, 1973. Other support: l) S Public Heahh Service. From the Medical Service. San Francisco General lloepital, arxl the F)tpart- ment of Medicine. Univenity of California School of Medicine. San Francisco. INTf?RRFI.ATI(1NSHIPS BF.TWFEN TTIE ItUMAN At VPOI AR MA(-ROPIIA(;F ANI) ALPIfn-1-ANTITRYPSIN 1luman alvrolar macrophales have been implicated as the source of an enzyme c.pahlr rrf causing emphyurna in o 1 anntrypsin drH.rrnl pat/cnls- but prt•n"us .Ilrnrt./r lu Lti ac srnh a sahslarKC have farlcd /lhr c.prrrmtnt.l wnr~ rrl..rtr.l twrr h~.+rvrr rkm..uslra4rs thAl nrrt rr/rly d.r thcsr cclss nor- ~n.t~t .nnu~r~ .1 k••r 1.y . I anl.llylnur, hul rl,rr con- tain Ihe inhibitor itself. As determined in this series of e.pcriments, alveolar macrophagcs lavaged from human lungs possess nlawrmat proteolylic activity sl p11 3 0; but pF1 profile curves also show what may he a secondary peak of activity at a shghlly higher ranRe, suRRestinR the presence of a second protcaw Proleolytic achvity, measured at ptt 4. 1. is inhibited by purified a-I-anturypsin Fluorescenl antibody studies show that the inhibitor is also present in normal alveolar macrophages. When such cells were taken from a patient with a honazyftosts deficiency of a-1-anlitrypsin, they displayed kss fluorescence a/ter incubation in autobtous serum than afler treatment in normal serum. Macra phars from normal wbjecls showed masimal fluoreseersce when removed from the IunR, and additional incubatioe with normal human serum did not increase fluorescersce. Cohew. A. a Thelourndo/(7lnirallwrsrirarlow 52(11):279)-2799, 1973. OtAer au pporf t U. S. Public Health Service. From the Medical Service, San Francisco General Ilospital. and the Specialized Center of Research and Department of Medicine. University of ('ahfornia, San Francisco. TFIE MOL.F('UI.AR STOICIIIOMETRY OF TRYPSIN INHIBIf1ON BY IIUMAN AI-PFIA-I-PROTEINASE INIIIBITOR Human a-l-proleinase inhibitor (a-1-PI), or o-I anlivypsin, is rnpomibk for more than 70% of plasma's inhibitory aelivity. It has always been aswmed that this protein interacts with proteinases at a 1:1 molar ratio Recendy, how- ever, /wo preparations of this inhibitor isolated by suhstantially drfferent pro- cedures, have been found to bind a Rreater proportion of trypsin. suggesting that one molecule of a-1-PI may h.ve multiple inhibitory sites. This report summarires the rnults obtained by utins two drRcrent preparations of highly purifind human a-I-PI to investigate the sloichiometry of inhibitor interactron with porcine Irypsin. In conlrasl to other reporu, one mok of a-I Pl wu found to inhibit two moks of trypsin F)ise gel ckclrophoresis indicates that this 2:1 compk is formed preferenlially even in the presence of free o-I Pl The mechanism responsible for Ihe inhibitory action of a I-PI is srill unknown and there are as yet no reporls to suggest whether any ut its Lxrrsds are ckaved during the rcactwsn Ihe investigators are currently sludyinR rhis asl.ect- as well as aUenspting to determine rf lartcr proteinases such as pla.min are Inhrhrled In the same stoichiomctric ratio as rep.uted here for trypsm, lohnaon, b A. Panncll, R. N. and 7'.ovir, /. elochrmlcaf anJ elophyrlrrJ Rnr.rrA ('onu.wnlrerr..nr 11( 1) iR4 1119, 1'!/J OtAer supporf: National hrNdutts of Iltallh From Ihe l)tpartmcnl of Himlxmntry. University ol (korgia. Athens 2S :t

IIIIMAN PAN(•RFA11(' FNZYMFS: CFIARA("i1 RI7.Al1ON trF ANIONIC Hl1MAN TRYPSIN Recently improved prraedures for purifying human pancreatic proteaset have made it possrhlc to dctcct an anionic form of human Irypsin, at well as to purify this protein and investigate its propertres The enzyme was isolated from acetone powdcrs of human pancreas by salt fractionation fotlowcd by ion eschanRe chromatoRraphy on SE-Sephades C-25 and on DEAF-Sephade^ A-S0. The preparation was homogeneous as determined by disc ekctrophoresn and sedimentatic.n equilibrium centrifuplion studies, the latter ako indicating an ettimated molecular weight of 25,1100 for this entyme. ('nmpariton of anionic trypsin with the cationic entyme shows a high degree of similarity not only in molecular werght• but also in the amino acid composition ut each pro• tein as well as pF1 optrmum ^nd ability to diftest casein. The ^monic Inrm, however. has only a weak cross-reaction with antilx+dies directed toward the caNromc entyme. and is much ksa stabk than that proacase It is also rapidly inhrbited by soybean trypsin inhibitor and, to a kuer, esteM. by porcrne Karal pancreatic inhibitor and ovomucoid; all three are poor inhibitors of carionic tryptin Mallory. P. A. and Travis. I eiochtmisrry 12(15) 2847 2851, 1973. OtAer auPport: National Institutes of Heafth From the Departmcnt of Bxschcmntry. Univenity of Georgia. Athens FUNC'11ONA1 AND BI(X'HFMICAI. FFFEC'TS ON THC I lIN(; FOI.LOWINO INIIAI.Af1ON OF Ct(iARET1E SMOKE AND CONSi1TUENIS I. NIGN• AND LOW-NICOi1NE CIGARETTFS IN MI('E •Tl+is study has esamined the eRects of chronic inhalation of cigarette smoke in two strains of mice, using lechniques prcviously developcd by these investigaton for rncasnrin8 pulmonary function in this specics The influence of aging was invesu8ated prror to delcrmining the cflccts of snxokc inhalatron •1At nicoline content of the cigarette and The duration of caposurc were varied in Ihesc e.pcrinvenrs No combination of these vari.hles showed a caucc atwl eflccl relatN.nsMp between cigarette smoking and chronrc abstrr.nuve pul. monary disease Daily inhalation of cigarette smoke for (ive ot rcn weeks did, however, cause the following: (1) increascd pulmonary resv%tance; (2) decreased functional residual capacity; (3) decrcascd pulmonary camplunce; (4) decreased tK1al volume; and (S) increased wcl weight of the Irmg relative to rcduced Ixwly wcrrhr Ihere was no change in pho+phohpid cruucnr of the lung the increase it .iImonary resnlance and dccrcau in lurxtrnn.l resrdual nKC>~nercunlcnl of htvcsc ~arclrtaar>,Ipllvcpdur•- cap•c',t~xr flesh Ihty were tion of esprxure, indicating that both are caused' hy a combination of the nicotine in parriculates and constituents of the vapor phax The decreased pulmonary compliance was elicited by inhalalron of fresh whok /nwke but not by the ps•vapur phase. This would indicate that the causahve factor is in the particulate matler, probably nicotine, because the appearance of decreased compliance depended on the nicotine kvel. The decreased tidal volume as well as increased pulmonary resislance, or bronchospasm, occurred more readily in ICR strain mice than in the Swiu slrain. Roth strains developed tolerance to bronchospasm after ten weeks of eapcnure. llure was no increase in fundronal residual capacity and, hence, no functional sign of pulmonary emphysema in mice that had been espoxd to cigarette smu-e for five or ten wecks. Avlado, U. AI. and Watanabe, T. Toskolory and Applied Pha.marototy 30(2): 18 S 200, 1974. From the Department of Pharmacolo8y. Univcrsity of Pennsylvania Schrpl of Medicine, PhJadelphia. FUNCTIONAL AND BIOCHEMICAI. EFFECIS ON T11E Ll1NG FOI.L.OWIN(7 INHALATION OF CI(iARETTE SMOKE AND CONSiITUEN IS. 11. SKATOL-E, ACROLEIN, AND ACE1 ALDEIIYDL° Daily esposurc of mice to the 8as-vapor phase of cigarette smoke for Ave or ten weeks has eliciled bronchoapssm and decreases in functional residual capacity. To help pinpoint the causative facton of these eflects. Ihreo con- stituenh of the gas-vapor phase, skatok, acelaldehyde and acrolein, were tested in male Swiss mice Oral ingestion of akatcde caused pcrtmonary con• reslinn that was manifested by an increase in pulmonary hemo8lohin content and a decrease in pulmonary phospholipids. There was no increase on functional residual capachy and, hence, no pulmonary emphysema in mice that had tither ingested skatole or been administered an inlrafracheal injection of papain The latter procedure caused decreases in pulmonary compliance and in the antl• Irypsin aclivity of the blood, both of which are coincidentally seen in human forms of pulmonary emphysema. Acelaldehyde, the second consutuenr of cigarette snsoke strwhed, when inhaled by mice for flve weeks c-used a tcduc• hon in (uncliunal residual capacity similar to that cncounrerc.l in mice e.- prnrd to the gAs vapor pluse of cigarette snwr-e Acrolcrn, she rhird cunqxxnKt whose eRcc'rs ^re hcrern reportca, causcd a reduclwrn its pnlmanary rumpbanre, but this effect is not seen in mice ealxned to the gas varyrr phase of cigarette smoke. Watunahe. T. and Avlodo, 1) At. Tuakolary and Applied PharmocoluRy 70(2):201 21W, 1974 From The Ikpartment of Pharmacolu6Y. Universvly of Pcnnsylvani^ School of Mcdicirx, Philadelphia 27 26

, t 1 r .+:.. ' Ll1NG CI-11. MI1(K'lIONI)RIA: RAPID OXlf)ATION OF (iLYCF.ROI.-I PIIOSPIIAIF BUT SI.OW OXIUAIlON OF 3-1-tYl)ROXYBl11-YRAI E Although Blycerol-l-phosphale (G-1-P) and 1 hydrosyhutyrate (1-HO8) are not uwal rricarhnsylrc acid cycle inlermediNCs, they can he osrJrred through the milochondrul electron traruport chain to drhydrosyacetonc-phos- phate (nIIAP) and 1 osyhutyrate (3 08) respectively. The oxidation reduc- tion process can scrve to transfer reducing equivalents from cyloplasm to mitochondria, simulrantously furnishing cytoplasmic nicotinamidc adenine dinuckoliJe (NAI)) lor continuation of glycolysis In additinn, the 1 HOB/1- OB ratio has bccn uscd to determine the -iytramitochondrial free NAI)11/NAI) ratio in liver cells and pulmonary alveolar macropha8es (PAM). Mdoclusndriat oxidation of G I P and 3-11OB, therc(orc, may be physirdo8ically significant not only in the conservation of energy by ottidatrve phosphorylatron. Mst also in the reprlation of rnttaholac utdualion reslricted by inlrace11u1ar cornpars- rnenution This cnmmunrcation reports the relative rates of G-1-P and 1 11108 oxidation in lung mrtochondria. Aa judged from oaygen consumption, lung mHochondri oaidrred G-1-P at a ratt (30-35 nmoks 0:/min/mg prolein) comparable to that of 2aso81utarate. or SO to 60% that of succinate. The 3 1109 oxidation rate was slow (6 8 nmoks 0-/min/mg protein)- correspond- init to 20% that of 2 oaoglutarate or 10% that of succinate. In addition to pro- viding NAn. G 1 P oxidation in lung mitochondria may also incrcase the livadahilrty of phospholrprds for oxidation and coupled energy production. The -slow oxidation of 111(lB might be related either to lack of an operative en- zynse system or to rtt inhibition by fally acid oxidation Althtxr8h PAM mito- chondna manifested charactcristres srmilar to those of total lung mdochnrxhia, it is not certain whether asl lung cell types have the same potential lor oaidiz- in& G-1-P and 3 11011 Muslafa. M. 0 and Crorr, C. E. Amcrkan Review of Rtrpiretory nirtait 109:301-10). 1974 Oflrer .u pporl: ll. S. Public Health Service. From the [)epartn.cnts of Biological Chemistry. Internal Mcdicinc. and Iluman Physiolo8y, University of California School of Medicine and California Primate Research Center. I)avrs. TIiE (iRANUT AR 1-YPF 11 PNF.UMON(K'YlF AND I UN(i AN11OXII)AN1 [)liFENSE It has long been known that type 11 ctlls, which covcr areas of the alvtolar surface no/ lined by type I cclls, contain larncllar inclrnions rcptcstnunt syn- thcsis sitcs (ot pulmonary surfacunt, the phospholrpid rich suhstarxt primatdy rtsponuble for alveolar stability More recently, howcvcr- three other rclatcd cnnccpls concerning the type II pncumonocyle have also been suRRc•.Icd: ( I) it parlrcrpalts rn lhc ,rnlury anJ rtparr prrrrsses nl p,Jnu,nary partn.hyrnal Insut; ( 2) it may I~r the pro[rnitnr of the m„re numtr„us type I alvri,lcr rprlhchal rtll%, anJ 1 1) Ihc rr1,0•livc .Japlrve rc+(xrnst Jelcrnunes, to some tslcn/. the susccl•11l„I iy „I I1,,,g p.rrnihy,ua to ucrJ.nl slrc%s Ihis editorial sunuuantes 28 I these three mutually complementary concepas as lulhrw. ( 1) As a reparative ccll, various csperrments have shown that prolJerauon of type 11 cclls occurs early in the coune of repair of Irm` injrny causcJ by a nwlrrplrcdy of irri- lants; and recent uttrastrsatural and cytokinelic stuJics indreate that type 11 cell hyperplasu occurs in o.idant rnduced lung damage. Ihrs response sums to be well developed within 18 bours of o.idant tsposurc. (2) Sequential cyto- kinetic and mutpholo8ical observatKins of lung repair altcr oaK).nt damage havc shown type 11 cells - hul not type I cells - in mitosis; morcover, alrcr alvcolar injury, labeled thymidine is initially founJ in type (1 cclh, only later in Uansi- tional cells, nJ flnally in type I cells (3) Reparative type 11 ccll proliferation coincides with the temporal appearance of lung tolerance to o.iJants A possihk mechanism of such tokrance was sugScstcd to one investigator by the favorable influence of Hluuthione ((]SII) on the susccptrbrlcty of the lung to o.idants. Also, consrderabk increases in glucose-6-phosphate dthydrotenase ((i6P1)) occur in lung lissue a/ler low dose- tolerance prcK)ucmg e.hnures of animals to o.idants lhe au8mented (i6P1) aclivuy prrrl+ably reflects increased activity of the hesose-nutrwrphosphate shunt. Recent findings show that low- dcxe oaidant e.posure. (kvels that caused noticeable type 11 cell proliferation) result in increased levels of GSH. Blutathiorse reducuse- other drsutflde re- ductases, and Blut.thione perosidase in the lung 24 to 48 hours a/ter the initi.- twn of taposures. The close, temporal relalion of the occurrence of biochemical and morphological changes and the o.idant tolerance development is striking In summary, reparative type 11 cells may well he considercd a part of the alveolar antioxidant defense system. Crou, C. E. A nndf o/ Internal Mtdl. inr 80(1) :109-4I 1, 1974. Other .u pport r National Inslitules of Iltalth. From the lJnivenity of California School of Medicine, Davis EFFECf' OF OZONE EXPOSURE ON LUNG M11(>CIIUNURIAI. OXIDATIVE MI:TABOI.ISM Although morphologic chances resulting from ozone (U.) have bcen rc- ported in lung muoclsondtia, the functional eQects have not been adequately investigated Ilcre the authors attcmpt to determinc rl Oj inhibrts lung cellular function by interacting with mitnchonJria which contain a high de8ree of un saluralcd lipids in their Ihpoprotcm memhtancs anJ are rhus susccpubk lu struclural YINI /urM1N)naI alltratMms by osrJirint a8enls RruJrs inJrcatc tha Os c.pcnure rndtrJ affects stvtrcd basic mdochondrr.l hu.chuns. ( 1) the rate of sufn/ralt: uswlation; (2) Ihc'rhddy to per/urm o.HLlrve Phusphurylatwrn; arKl (1) cerlarn pcutxabilrty characterrstics lung m.uo.hr,nJna /IUm young heallhy rats e.poud to 4 plxn O, for (rrur houn, a hrgher level than cepecrcJ in ambient almospheres, were prcparcJ with the usc o( an rs„lomc huffercJ aucrusr mannunl mtdwm (-omparrJ to cuntrrd prtP.ralruns Ihc c.pmrJ mdrKhnnJrra nurult.NtJ a Urwer uityRtn cunaumpunn rare (.11 agnarnts uf the tcslru.tary charn htm8 .uutpnhlc Io l)r c.ir,.ure) anJ Jrnunrshcd cfh cicncy of coul,IcJ phusl,hr,ryl.hun Mcmhr,rne pcrnrcahrlrly rncrtacJ cunuJ 29

erahly. 7hrol ( Sit) Icvelt dn+pped ?OR, indicating oxidation of mtroihondrial protein -S/f 6roups, an mrprrrlant mechamsm of orid.nt damaRe I ipvl pero.r- datron was evMknt in rnr.r lbere seemed to he smular, d smaller. cl,+nRes at lower Os kvels (helow I ppm) Mn these were not cons.<fered sign.fic.rnt in the lesser numher of animats studied. Yet, indirect effects such as 1) rnduced release of lysosomal hyJrolases in the lung may also potentially contrrMue to lung mitocho.xlrial damaRe in riru. lhe authors are currently inve.tikatrnR the possibility of favorably modifying (lrinduced damage with various anliosi- danls protective against thiol oxidation awd lipid pero.idation in vitro (such agents have proven helpful 1o certain plaets against photochemical c.alants). Nevertheless, they do not fcel Iha1 at thia point in time the evidcncc that (>ti directly aRecls lung mitochondria /w vlvo is indubitable. Mustafa, M O., De Lucia, A. 1., Crou. C. E.. York, O. K. and 1)unRworth. D. t-. Clrerr 46(1) :16S I ttS, 1974. Other .urport: U S Public Heallh Scrvice-National Inqilutes of Health. From the Departments of BioloRical Chemistry. Internal Medicine and 1luman Physiology. School of Medicine- tkpartment of rathology. School of Veterinary Medicine, and California Primate Reaearch Center. University of California. Dw'n. FFFFCI S OF SIIOR T.l FRM OZONE EXPOSURE ON I.t/N(l MTT(X:FIONURIAL OXII)ATIVE ANE) ENERGY METABOI ISM This'sludy demonslrales that mitochondria in lung cells may be impor- tant targets for orone (O0 interaclions, and that interference of O, with lung mitochondrial o.idative and energy metabolism may significantly contribute to the overall lung damage that results from acute esposure of aninuls to this osidant Shorl4erm, high kvel Os eaposures (2 ppm (h for ta hours, or 4 ppm for 4 houn) deprest lung mitochondrial Os consumption, coupled pM)aphoryla- tion, and respiratory control in rats and monkeys. 1 ung mitochondria from control animals were relatively impermeable to added reduced nicotrnamide adenine dinuckotide (NADH), bu1 those from esposed animals shu..ed in- creased permeability as judged from NADII oxidation which was a threefokd increase over the conlrols. The depressed pulmonary milochominal functions observed in esposed animals may be related to atteratron o( membrane permea- hility due tn lipo) oxidation arKd to inhibition of respiratory entymcs (Aehydro- genases) due to oxidation of functional thiol group.. Mwtala. M. O and Cron. C. t:. Archivr, oJ Dlorheml,r.y and Riophyilct, 162: SlSS 594, 1974 Other .upport: O S Puhlic Health Service From Ihe Ikpartrnentr uf Ifittlusical ('Lemntry, (roern.l MeJw rra. .nJ Ilu.oan I'hynnltrRy. S.h-Md nf Mcd,cine. and ('aldornu Prinutc Rexauh l'cntcr, (Jni- vcnoty trf (-ah/urnu. I).vis )O I THE ROLE OF PULMONARY SURFACTANI IN EIEAt: fN AND DISkASI°_ In this carefully considered overview of the role and importance o(pul. monary surfactant. the author crrtically surveys Ihe historical h.ck~rousd, established measurement lechniques, data defkiencies, and pressing research needs of this intriguing lung subslance. It has long been kr.own that the ter- minal bronchioles and alveoli of the lung are lined with a substance, pul- monary surfaclant, that markedly reduces surface-active forces, stahdir.ea the dyeoli and prevents ate"ectasis. This lining compka consists predominantly of saturated pho.pholipds with dipalmitoy( lecithin (DPL) as the major active component. An adequate amounl of'active surfactant is essential for normal IunR funclion. EEowever. the role of surlacuM in pulmonary disease is diRicult to asaess because this substance cannot he quantitatively measured by direct means. Currently available techniques for measuring surfactant are lung oorw- pliance, surface activity of lung estracts and biochemical measurcmenu of lung photpholipids. (n the presence of patholosic changes, however, thex qualitative indirect measurements do not allow us to distinguish a primary de- fkiency in surfactanl from a secondary one, from inhibition of activity or from inadequate sampling. The lung is rich in DPI-, Mrl the /raction of total lung DPI. which is surfaclant is not known Although the large alveolar cell is probably the source of the lining compka, there are two other cells is the terminal airways which actively synthesize DPI., the Clara cell and the mono cyte-macrophate. Changes in metabolic activity and numbers of these cells in pathologic sutes also need to be known in order to evaluate properly allera• tions in lipid metabolism in disease. Therelore, the author concludes that the eaact rok of pulmonary wrlactanl in disease is, a1 present, essentially un- knowo and speculative. Escep Eor fetal immalurity, there is no known clinical condition in which a primary deficiency in surfactant has been cstablishcd lEowever, the author believes that a secondary deficiency in activity of the lining complea does play an important role in the pathoRenesis of many dr• ease slates, but in which ones and to what eatent is not known Similarly, a primary deficiency in surfaclaM activity will probably be uncovered in lime. In conclusion, a few eaampies are presented to denxtnslrale and emphasize the currenl pitfalls in interpretation of available data. There is an urgent need to develop direct quantitative measuremenls for pulmonary surlactant and mwe critically ctxrclate the palhobgic, functional and biochemical derangementa in the diseased lung. Only then will we know the proper role of pulmonary .ur- faclant in disease. Nidrn, AlDorrt N. In: lohnalon, R F(cJ ): Pufmonary ('are, New Yurk : (3rune and S/ratton. Inc., 1971, pp a)-101. • Ofher support: U. S Public Ileallh Service. From the t?epartment of Medicine, 1)rew Postgraduate Medreal Sch.al, t.o. Angeles 11 I

RFVFRSIBI E hAMAGE OF= RAT UPPFR RFSPIRATORY 1RACT ('Al1SFD BY C'I(;ARETTE SMOKE Because of the need to determine the specifk reaction of tlse masiltary gland and sinus to an etptrimenlally-induced environment, this pr,-liminary in- vesli8ation was designed to clarify: (I) the eRect of whole ci8aretle smoke on the masillary epithelium and gland; (2) Ihe progress of morphaL.BK changes in Ihe epithelium and Rland as a function of smoke dosa6e; and 11) the mor- phologic chantes in the epithelium sd gland during the perind of recovery. Adult SpraBue-Dawky rats were eapoeed to whole cigarette smot e for diRer- tnt periods of time and then saeriAeed at various slages of recover,,. As studied under light microscopy. histologic prep.rations of masillary sinuset and glands showed aeveral changes: (t) the masillary epithelium lost colursnar ciliated celh and hypertrophred: (2) Iht submucosa displayed evidence of an acute in- flammatory reaction: ()) in addition to other morphologic mod fkations oc- curring in the masdlary 81and, individud goblet cells containing ar id mucosub- stanccs diRerentiatcd within new maadlary epilhelium; (4) :arravauutar IympAocytes, polymwphors,rckar kutocytes, and macrophaRes migrated in great numbers from the subrmttosa to the sinus lumen by way of the epi- thelium; (S) some migrating cells invaded the lumina of eacrelory ducts; (6) the masillary sinus contained a massive amount of pus; and (7) rsicroahceyaes were scattered throughout the epithelium of animals esposed to smoke for • longer time. In spite of the morphologic changes induced by the caperiment, normal structure around the matillary sinus was retstabinhcd quickly after cessation of srnolin8 and throuBhouW roco.ery. Vidic, B, Rana, M W and eAarat. e f). ArrAlvtr o/ Orolwrntol,rty 99( 2) 110 I I l, 1974 OtArr support: National lnshtute of Menta) Itcalth and 1/. S Public Ilcalth Ser.Ke. From the Depariment of Anatomy, (leor8etown l/niversity School (if Medicine. Washington. 1). C, and the 1)epartmcnls of Anatomy and Physiology. Saint Louis University School of Medicine. St. Louis. CNANC)PS IN TRACNEOBRON(1f1AL CYTOLOOY NOTED DURINO ANFS111/3SIA (e this tracheobronchial cytologic study, snxan from 4,571 patients un- der8oin8 general endo/racheal anesthesia were screened for the early diagnosis of bronchopulmonary tumors In addrlion, one or more of the following meas- urements - to1.l nuclear score, percentage of goblet cclh, total cellular scrwe, percentage of muhinuckated eells - was made, accordin8 to the particular study in protress Results fcll either under the heading of changes in cytology caused by anesthesia or chanRes in cytology coincident to anesthcsia, and the following plserwmena were ducovered: (1 ) sieniflcant cytomorphologic chan8es occur in the epithchal cells of patients who inhak dry nesthclic gases for longer than one hewr; (2) the tracheerbronchial epithelium undtrjnes vanations rn morpholngy which resemble changes found in the endometrium during the memuual cykIr, (1) mcresttd numhen of muttusuclesteJ cJuteJ epitheGal cclls and a generally higher degree of multinrklc.twrn are found in patwnu with etlrathorac.c mali8nancres; (4) cytomorpholosrc damage in smolen prc- cedes reductions in lung function; and (5) tracheuhronchial washmp may be used to diagnose inhalauonal injuries sustained in fires and, of course, to dr- cover unsuspected mal 8nant condilions of the lung and bronchus. Clialon, l. New Yorl State lournal o/ Mrdlcine 74(12):2185-2189, 1974. Other supportt National Cancer Institute and Arnerican Cancer Society. From the Departmenl of AncslhesioloLy, Albert Einstein ('ollcite of Medicine of Veshiva llnivenity, 'The Brone, N. Y. TRACHI:oBRONCFIIAI. CYlOL(X:IC CHANGF:S FO1.1 OWIN(7 LOWER AIRWAY TIIERMAI. INJURY: A PRLLIMINARY REPORT Esfoliative cytology is a technique which might provide a relatively aon- invasive, atraumatic means of making a definitive duRnosis of the thermal damage to the lower airway caused by smoke inhalation. In this paper, t!e r.- wstigaton describe Iracheobronchiai cytology from nine patsenls with over- whelminR clinical evidence of inhalation injury sustained in flres Cytology was grossly abnormal in two palienb; there was moderate evidcnce of damage i. Icwr nwre cases: nd smean from the lau three patients were normal. Thcre was a positive cqrrelalion between cytologic damage noted in smears and scverity of clinical flndings during the first hospital day. Sequential scrial cy• tnlogy in two patients reflected the course of the ksion during therapy. These results appear very promising both from the diagnostic and prognostic points of view. lhe method is simple, praclical, and inespensivc; perhaps it should be evaluated at other huspitats. Ambiavatar, M., Chalon, /. and Zargham- 1. The lourna/ o/ Trauma 14(4):280-289, 1974. From the Ikpartment of Anesthesiolop, Albert Einstein (-olk8e of Maficiae of Yeshiva University. The Bront, N. V. 111. Ilrart and (art•nlafinn CORONARY BI l)nI) FI OW AS.SFSSMt:NT WIfH XF.NUN ANI) R1/811)IUM -Ihis paper presrrNs a clinical tcvicw of the mclh(wls uung rssXt and "Rb for the dclcrminjhun of cor4aury hkxr.l flow Alhh aiKh cnronuy Nood 8ow has heen mcasurcd in man for over 25 yean, an iJcat cl1mcally applicable 32 33

method has not been eslahlr.hcd yet The hes( meavue availahle mrw, bascd on the authors' tsperrentc. seems tn he the "Rts coincidence connrrng tech- ruque snsce it nuastures nutrrtUonal flow throuch the whok heart rcR.rrdlcss of changes in myocardul fluw JnrriMuion. ard since rt is a nonmv-rvrve technique. 1 he main linntauon rsf Ihis system lies in its inaccuracy (or al.,ulute quantufi- calwrn at high flow rates AlthouRh the determinatiun of nurrrtional hlrrwl 11ow by thc rubidium clear.rncc technique underestimates very high flow rates. it reflects an accrualt estrnrate of directional changes in myocardial blood nuw. In the atnon mcthod, the major problem arises when flow is not uniform throuRhoart the heart. as is certainly the case in coronary artery disene 'Ihen the calculatron of flrrw from mullieaponewlial decay curves is pruhicnutrc. Another Jiffscuhy is the dependence of the numerical flow value on the tissue- bhttxl partition cncfTicient and ils ponibk changes in a diseased state An addi- IN1naI dnadvant.Re is the requirement of catheterizatusn of the coronary artcr- ies; howevcr, thrs permits separation of Row through ritht and left coronary circulation and direct correlation of flow values with anatomrc varialions seen by coronary arteriography. Pachinger. 0 M.. Tillmanns, ll T. and Blng, R. !. Srnunars irs Nuclrar Mrdicinr 3(2):I31-131, 1973. Other support: Hoover Foundation and the Wright Foundation. From the HuntrnRton Memorial Ilospital, Pasadena, Cal , and the University of Southern California. I os Angeles Sflll)I!_S ON 111F (OR(1NARY MI('R(X'!R('l11 ATION BY 1)IRHCT VISl1A1 llAfION Since little had been published on phasic Row in the coronary microcircu- lation, these sturlres focused on (1) the anatomical pattern of the coronary mKroclrcula11tN1 and rts relationship to the direction of flow from one capillary to the other, and (2) phasic flow in the coronary rmcrocirculation To caplore these Ihinp, the coronary microcirculation of the cat was nburvcd by trans- illumination of the left auium. In most insaances, high speed cinematography was used; red cell velocity was determined by frame to frame analysis during both phases of the cardiac cycle Countercurrents and asymmetric capillary arrangemenl were found Ihrs has far reachinR importance for the oaygenarinn of the heart muscle and makes doubtful previous cakulations for the utyRcn transport from the capillary to the surrounding tissut Rccrurtment, dcnonnF an increase in the number of capillaries, was observed with a rise in pafusion pressure and following the admimstralw.n of nitroglycerin It can occur in the presence of sutore6ulalion l)cM1nite patterns lor red cell velocity in the capd- taries ol the left atrium emerttd Nicotine and nitroglycerin had lrllle eRecl nn rrd ttll vchxity Ilowtvtr, ftA{owinR hemorrhage alonc, anJ altrr Iht ad- nrrmrlr.rum rd rurrr,Rlyitrrn frdlr-rnR henurnh.Rt. m.rkeJ thanFr•. in the rrd rrI! ..I,.rty ..rrr vrn llrrrrnrrh.lt -meJ • m.rlcrl dmunulrrvr. whrn frJL L..r.l 1.~ n~r...rr„r,„ ,r rr.,rlr,A rrr . nurlrJ rn,rt~a rn rrJ crll vcl.xtly .tr,l.,r. . Lr.,.r...~ .~..,..,. ....... .r, I Binl, R. /. ez al. In: Maseri, A (ed ): Myocordia/ Blnod F'lor• in Man Afrrhu.lr and SitniF- contr rn Coronary 1)israrr. Turin: Minerva Medica, 1972- pp. 21 )3. Other aupport: American Medical Assrscialion, 17(vnvcr Founduion, Los Angeles County Heart Associalion, and the Norris Found.hon. From IlunlinRton Memorial /lospital and California Institute of lechnolo(ly. Puadcna, Cal , and the University of Southern Cah(ornia, I.o. AnRetcs. LIPID METAB(11.ISM IN HUMAN CORONARY ARTIiR11:S Mechanisms of the formation and.upsake of fipids into Ihe perfused hee- man cornnary artery are discussed in this paper. For the caperimentu per- formed, human plasma was used as a per(usate; '11 labcled cholesterol and s'C-acetale were added. Cholesterol was dispersed in the mcdium by sonica- lion. The results revealed lssl both alherosckrotic and normal coronary arteries of humans faikd to synthesize cholesterol and cholesterol esten. but thal they did incorporate "C-acetale into other lipds. Similar readts were obtained in human saphenous veim perfused at arterial pressures In conlrast, uptake of cholesterol from the perfusion fluid was demonslrated in atherosckrouc and normal human coronary arleries, as well as in human .aphenous veins In this study of lipid melabolnm, results illustrate that under the conditions of Ihese esperirnents, normal and atherosclerotic human coronary ancries, as well as human saphenous veins, synthesize free fally acids. IriRlycerides, and phcx- phtdipids Cholesterol and cholesterol esters are not synthesized, but enter the arterial wall by insudation or imbibilion only. Bint, R. J. N al. In: fMalla, N. S(ed ): Rerenr Advancer in Srrdie: on ('ordrac St.ucrurr and Mnebofizrn; Vol !- Myocardial Mrra6olir.n, Ballinwre: University Park Pres., 1973, pp. 33-Si. Other support: Noover Foundation and the Norris FourKlatwsn, From the Iluntinglon Institute of Applied Medical Researth, Hunlintton Me- morial Ilospiul, Pasadena, Cal., and the University of Southern ('alifornia, I us Angeles. 1,1141) MHfAB(111SM IN PI'RI:lISl1) ItItMAN NONA111FRO S('l 1iROI l(' C(1RONARY AR Il?RII S ANU SAPIII-NUUS VFINS 'Ihia study of Irpid synthesis .m/ cholesterol uptake in human nonathcro sclerotic corunary arteries and per/uscJ saphenous veins confirms previously eapressed opinions that there is no chuleslerol synthesis in human arteries Wtlh this series of espenmenls, the anrhurs dcnxinslrae that nenher normal nor atherosclerolie human coronary arteries can synlhtsize any cholesltrol from acetate, anJ that h.qh .rc capjbtc uf pnducmt only small answmts of ehoksteral esren (-MAcsterol uplalc, moreovtr. is identical in nornul and theruxkrutic arterncs, leading Lr the trrr)clusron thit the large amounu of 35 14