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Council for Tobacco Research

1977 Report of the Council for Tobacco Research-U.S.A., Inc. [St]

Date: 1977
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G~2 ~ o0 1977 KI:POR•r t3 of TIIE COUNCIL FOR TOI3ACCO IIUSEARCII-U.S.A., Inc. TIiE COUtiC1L FOR TOBACCO i(!'.SEAIt/:1t-U.S.A., Inc. 110 Eue[ 59t1a Strcct, New Yurk, N.Y. 100'l'L
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. SCIENTIFIC ADVISORY BOARD HANS MEIER, D.V.M., Dr. Mcd. Vct., M.R.S.H. M ~ Senior StaB Scierttist 00 to Tbc Council for Tobacco Rcscarch-U.S.A., Inc. a: of Decembcr 31,1977 SHELDON C. SOMMERS, M.D., Chairman Director of l.aboratories. Lenox Hill Hoapital i Clinlcal ProJasor o/ Pathology Collegc of Physicians A Surgeons of Columbia University New Yoct, Ncw Yocf Thc Jackson Laboratory Bar Harbor, Maine LEE W. WATTENBERG, M.D. Professor of Pathology DePartmcnt of Laboratory Medicine and Pathology Uruvcrsity of Minnesota Medical School Minneapolis, Minnesota ~ RICHARD M. BINO, M.D. Director of Cardiology and Intramural Medicine Huntington Memorial Hospital, Pasadena, California ProJessor o/ Mediclne University of Soutbcrn California School of Mcdicinc Los Angeles, California JQHN P. WYATT, M.D. Director Tobacco and I icalth Research Inrtitute University of Kentucky Lexington, Kentucky F-- U JOSEPH D. FELDMAN, M.D. Head, De nt of Immunopathology Scripps inic and Research Foundation Scientific Staff of The Council La JolIa, California WILUAM U. GARDNFR, Px.D. Scientific Director, The Council for Tobacco Rcscarch-U.S.A., Inc. E. K. Hunt Professor oJ.tnatomy (emeritus) Yak University School of Medicine New Havcn, Connecficut WILLIAM U. GARDNER, PH.D. Scientific Director ROBERT C. HOCKETT, PH.D. Research Director DONALD I i. FORD, Pu.D. VINCENT F. LISANTI, D.M.D. ROBERT J. HUEBNER, M.D. Associate Research Director Associate Research Director ChieJ, Laboratory of RNA Tumor Viruses National Cancer Institute JOHN H. KREISIIER, Px.D. DAVID STONE, Ptt.D. Bcihcsda, Maryland Associate Research Director .itssociatt Research Director LEON O. JACOBSON, M.D. Director, The Franklin McLean Memorial Research Institute Regenstein ProJe.uor o/ Biological Sciences Univcraity of Chicago ChicYgo, Illinois HENRY T. LYNCH, M.D. Professor and Chairman Departaoent of Preventive Medicine and Public Health Creighton Universiiy School of Medicine Omaha, Nebraska
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r , Introduction coNMErrrs Introduttioo . . . . . . . . . . . . . . . . . . 5 Abstracts of Reports . . . . . . . . . . . .` . . - . 6 Canccr-Rclatod Studies . . . . . . . . . . . 6 Tbc Rcsp'tratocp System . . . . . . . . . . . . 22 Ncart and CLrculatioo . . . . . . . . . . . . . 34 Ncuropharmscoioyy and Pbysiology . . . . . . . . 44 Phuusaco~ and Biochcmistry . . . . . . . . . 49 Immuno1ogy and Adaptive Mcchrnistns . . . . . . . 54 P,pidcmioloa . . . . . . . . . . . . . . . 59 Mis<xllanoaus . . . . . . . . . . . . . . . 63 Active I'ro}ccts . . . . . . . . . . . . . . . . . 64 Compkted Projiects . . . . . . . . . . . . . . . . 73 Indcz o( Principal Investigators . . . . . . . . . . . . 83 Iasdcx of Senior Autbors . . . . . . . . . . . . . . 84 As its quartcr-century mark approacbcu, The Council for Tobacco Ro- xarch reviews its history with a scn.e of jrati&atioa over progrtss in several dircctions, sonic dcgroe of Irustration in othus, and a considcrabk mcastut of optimism for the future. In the past 24 years. Tbc Council bas provided substantial support for what may well be the world's most eztctuive non-governmcntal rescarch pro- gram conccrning tobacco use and health. Tbe basic policy adopted at the outsct has bccn maintained, that is, to platx tcsponsibility lor the direction and guid- ance of Ihe rcxarch c/fort in the hands o/ a Scientibc Advisory Board, and to havc thc rescarch conducted by indcpcndcnt invatigaton in their own insti- tutions. 2icnce, whatever contributions to acicntiBc progress may be attributable to The Council's program are due very largcly to the distinguished men who have served on this Board. With major emphasis upon cancer, heart diseases and chronic lung ailrncnts, ttx Board has consistently counselled sirong attention to the multiple steps and stages in the pathogcncsis of thcse aging-associated "constitutional discases." Basis for this emphasis was a conviction that rapidly developing new techniques and concepts in biochemistry should provide powerful tools for elucidating Ihex multiple steps and staga, with a reasonabk hope of short-circuiting or blocking one or more of them and thus prcventing the discase or delaying its onxt. By the end of 1977, this program had produced more than 1,500 fcports and articles in accredited acientific journafa by scientists acknowledging Council support. Thcir ultimate sijrtilicance in relation to the goals cannot be fully appraised at this lime. Tbe mosaic is complex and emcrgin= pictures arc still far from clear. It is frequently stated that, historically, many of the most important ad- vanccs in mcdicine have been made by unprsdicubk empirical discovery. Yet, in thcse days, such discoveries arc most likely to be made by akrt individuals in the course of systematic studies juwSed by a good rationak. Thc maintenance of scientific cnvironmenu •cooducive to such discoveries is, thcrefore, an im- portant responsibility ol any jranting orjanization. The prescnt report contains abstracts of manuscripts publishcd in 1977 that acknowlcdgcd Council sponsorship. Thc abstracts speak for themselves. They illustrate Council policy that grantocs alone shal/ be responsible for dis- ciosing their rescarch findings in atxcpted medical and acienti6c journals or before medical and scientific organiutans. The Council is continuing its program of support to indepcndent research- ers in bio-mcdicine with its original goals. 5
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. r Abstracts of Reports Following arc abtracd, approved by the autbors, of reports on new rescarch acknowlodtiag w+ppoit frorq The Council that have appcarcd in scien- tiBc journals ainoc pubiicatioo of tttc 1976 Report. The name of the recipient ib in italics. The abstracts aro srouped under the.c headings: 1. Cutcer-Related Studies, 11. The Respiratory System, 111. Heart and Cinculation, IV. Ncuropharmacology aad Physiology, V. Pharuucology and Biochemistry. VI. Immunology and Adaptive Mochaniuns, VII. P.pidemiolo=y, VIII. Misccllaneotu. I. Catscet+Related Sttrdiei REPRESSIBLE AND INDUCIBLE FORMS OF DIM kT.T'HYLNITROSA1vtiNF.DEMETHYLASE I`(ak SprasuoDawky rsts and inbred strains of male mice were used in this study of dimothylaittnsamioo (DMN)-0omcthylasc with respect to kinctic cbarsdaitttia and rsspoara to Inducer pretreatment; atrain-0ependencc of rc- pratubility and iaducibility was also iavcstlgated in the mice. Results show that two cnsyme forms, with diffcrcat kinetic cbaracteristica and opposite responses to iw vlro enzyme inducer pretreatment, underlie hepatic DMN-demothylase ac- ti.ity. In kinetic uudia, detcnoinatioo of the Hotstce plot of DMN-demcthyl- arro using a DMN substrate cooocntration range of 0.5 to 200 mM yields three Intcrsotiing line segments from which widely different K. and V_ may be ca1cvlated. Identically p.ucrncd Hofsteo plots arc obtained with rat and tttouw postmitocLondrial wpcrrutant frauions, as well as with the isolated microsorrwt The lorv-sub.usia-ranye line sejment (0.4 mM) and the high- substrate-ranga se$nxat (50-200 mM) corrapond, in both the rat and the toouao, to two ditfcrcot entymatic fonns of DMN-demethylase (DMN-dc- mothylaao I aod U, rtspoctivcly) wbich have the following diffcrent regulatory charactcrutica: (1) Pretreatment of rau and tnicc with the polychlorinatcd bipbcnyl. Aracloc 1254, bciap about repression of DMN-dcnxthylase 1 and induction of DMN-daaethylue 11; both responses arc stronger in the rat than in the mouse. (2) Pretreatment with phcnobarbital rcpresscs enzyme I in both spocics; bowcver, it induces enzyme 11 only in tho rat. (3) Prctreatmcnt with 3- mcthykho7anthreao wbstarstially rcprcsscs DMNdcmcthylasc I in the rat only; it is itscIIective in sitni8cantly iaducocins any other enzymatic activity. Impor- tantly, the acnetically distincx nature of the two forms of hcpatic DM N-Jcmcthyl- aso is suggested by the substantial diAcrences in the urain-depcnckncc of the repressibility of enzymo I and iadueibility of enzyme 11 in a series of nine in- bred strains of mice.'IDe mouse lung eontaias only the induciblc-type of DMN- demctDylasc; howevcr, the rankint of inducibuitics is diSercnt from that of bepat:c enzyme II. Arcar, J. C. cr al. Zciuehrifr /ur Krcds/orchwny wtd 1l'linixlu Oniologie 89:181-199, 1977. 6 Otlser srrpportf Natiooal Caacer Itratiuu.. u From the Se.mcn's Mcmorial Research Iaboratory. U. S. Public Health Scrvic. V Ho.pital, New Orlcans, and tLe Dop.rUatat of >•tediciot, Tulane Univreraity p( Medical Center, New Orleans. ONCO-DEVELOPMENTAL Af.KAI1NE PHOSPIfATASE ISOENZYMES Previous studies havo shown that tumor alkaline phosphatases have their counterparts in relatively early stagcs of human dovclopmcnt. For ezampk, the chorionic phase I isocnzyme in 6-]0 week tropboblast microvillar tissue coa- sisu of two heat-scnsitive, f.-homoarynino-inlhibited bands. The fastcr ono, 9F band A. exhibits none of the antijenic dcterminanes known for various alkaline lX phosphatase isoeazymcs and the slower one shares determinants of the liver F.,_ type. After ten weeks, alkaline phosphataae of the term placental type appcars V in syncytiotrophoblast ulls, and the chorionic type becomes much less promr- rxnt. Further evidence presented here indicates that the chorionic (6-10 week) tissue isocnzyme lacks the antigenic determinants of term placental isoenzyme. Still more evidcnce suggests that the counterpart of chorionic band A is to be found in the tatis, leading tbe authors to question whether this band is pro- duccd by teratocarcinoma of the testis or by canccr of the lung. In the present study of ncoplastic transformation in relation to oncodeveloprriental gCae ex- pression, the lracheobronchial tree of humans with cancer of the lung was chosen as the model, since all transformational stages from basal cell hypcr- plasia to carcinoma in siru can be expected to be found in the nontumor arr.as. Significant levels of urciroembryonic antigen (CEA) and human chorionic gonadotropin (HCG) were found in extracts of bronchial epithdial cells from non-ncoplastic as well as neoplastic areas. It may be, therefore, that the onco- dcvclopmental gerxs for CEA and HCG arc turned on early in transforming cells and penist in malignancy. Fithman. W. 11. rr al. In: Fishman, W. H. and Scll, S. (cds.): Onco-devclopmanrol Grnr Erpreuion, New York: Academic Press, Inc., 1976, pp. 165-176. Other support: National Cancer Institute. frrom thc Tufts Canccr Research Centcr, Tufts University School of Medicine, lknton. I't1ENOTYI'1C ALTERATION OF ISOENZYME PROFILES OF ALKALINE PIIUSPHATASE IN HeLa TCRC-1 CELLS GROWING IN IMMUNOSUPPRESSED RATS Whcn grown in culture, the human cell line, HcLa TCRC-l, produces the hcat-slrblc carcinoplaccntal Regan isocnzyme of alkaline phosphatasc esclu- sivcly. ]fowcvcr, as this paper describes for the first time, a profound altcratioa takcs place in the isoenzyrne profile of alkaline phosphatase when HcLa TCRC- I is grown as a solid tumor in immunoauppresscd rats. Acrylamide gcl clectro- phoresis rcveals a decreaae in the Regan isoenzyme with a simultaneous appear- ance and increase in new fa.st-moving isoeozyme bands. After 30 days of 7
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growth ln vivo, no Retaa i+oeazymo )s visiblo and a siagle faat-moving iso- enzyme, band is the dorttinant iaocasymc. This new cnzymc form ia idcntillcd as the oncountsionlc (FL) isocnxyrrse Rnt characterizcd in the FL amnion celi lirse and iater in a hcpntaaa paticnt Upon tramfcrring the tumors back to cul- turr, the cells rcjain ilrek orisinal Regan phenotype aftcr one to two weeks. Nevcrthc1ess, cvca though the exprewioes of the Regan isocnzymc is rcatored, thc HeLa TCRC-1 celts' lsoaszymc expreuion has been changed, since this ccll line alter being paucd tbrough an immutsosuppressed rat is able to rc-express the otscoarrsniotic isocazymc at ccrtain cell dcnsi:ies in culture. Such growth of hurnan canccr ce/ls in itutnunasupptessod animal hosts may provide a modcl systtm for the study o[ carcinoXcnic expraaioa. Sia.-er, R. M. and Fishrrsan, W. H. In: Fishman, W. H. and Sell, S(eds.): Onco-devcfopmenraf Grnc Eaprctrion, New York: Academic Press, Inc.. 1976, pp. 177-184. Other sapportt National Cancer Institute. From the Tufts Canccr Research Ccnter, Tufts University School of Medicine, Boswn. CARCINOPLACENTAL ALKALINE PHOSPHATASE: BASE LEVEL AND HORMONE-INDUCED ACTIVITY ASSOCIATED WITH EVENTS IN THE CELL CYCLE The Regan isocnzyme (a carcinoplaccntal gcnc product first discovered in a bronchogenic cancer and subsequently found in a variety of other human carscus) is a placental alkaline phoaphatasc isocnzymc which is also produced by the HcLa cell model system. HeLa TCRC-1, a particular cell line which is rnonophenotypic with respect to the Regan isocnzyme, is cxamined in this study of prodniaoloaa rcgufation of alkaline phosphatase as a function of evenu in the cell cycle. Results show that DNA synthesis is not required for hormone induction of the Rcjan lsocnzyrne since induction occurs in the presence of hydroxyurca, a specific inhibitor of DNA synthesis. Additionally, when partial- ly synchrooizcd cells are allowed to leave the S period prior to hormone treat- ment, and hydroxyunea is added to prevent cells from enicring the next S period, hormone induction of the Rcgan isocnzyme is still otncrvcd. This indi- utes that initiation of expreuion of hormone-induced carcinoplaccntal alkaline phosphatase occurs prior to the DNA synthetic phase of ttic ccll cycle. -lhis finding can be harmonircd with previous uudics on other ccll lines by hypothc- sizins a two-step mcehaniun of hormone action in which the first induction step requires DNA synthpis, while the secorKd step, which is more closely rclatcd to transeription and translation aequencea in the Gt period, does not. It is this aeeatd atep whieh is functional in the TCRC-1 cells. Singtr, R. M. and Firhman. W. 11. Di/Jerenriarion 5:127-132, 1976. Other support: National Cancer Institute. From the Tufts Cancer Rescarch Center, Boston. DEVELOPMECI-(-AL PHASE-SPECIFIC ALKALINE PHOSPHATASE 1SOENZYMES OF HUMAN PLACENTA AND THEIR OCCURRENCE IN HUMAN CANCER The appcannce in tumors of a dcvelopmcntal protein antigen such as Regan isocnzymc, a placcatal-typc alkaline pbosphatasc, may indicate the to- exprcasion of a gcne set characteristic of a particular event in early dcvelop- ment. Information of this sort may help dassity stages in neoplastic protrtis- sion. This preliminary communiutloo describes alkaline phoaphataac ckctro- phorctic patterns charactcristic of thrce phases in early human trophoblastic developrnent. Thc Phase 1(6 to 10 weeks) pattern consists entirely of two heat-senaitivc. Lhomoarsioine-inhibitcd bands, the slower one possessing anti- gcnic detcrminants of the liver-bono type of alkaline phosphatase and the fastcr one lacking any of the known alkaiinc ptsosplutase antigcoic determi- nants. Tlsc Phase 2(11 to 13 weeks) pattern shows a mixture of Phase I and Phase 3 (14 to 16 weeks) isocnzynx components, the latter of which has two isocnzyme bands with the characteristics of term placental alkaline ptwsphatasc. These three patterns of devclopmental phase-specibc placental alkaline phos- phatasc correspond, in order of appearance, to non-Rejan isoenzymc, a mixture of non-Rcgan and Regan isoenzymet and Regan isoenzynx as are found in a variety of human cancer tissues. Examples of each phase's occurrence in human cancers arc given, arx] in this way a guide is provided for interpreting tumor alkaline phosphatasa as retkctions of the activation of phase-specific tropbo- blastic gcoes. Fishman, L., Miyayama, H., Driscoll, S. G. and Firhman, W. H. Canccr Rctearcb 36:2268-2273, 1976. Other support: National Cancer Institute. From the Tufts Research Center and the Department of Pathology, Tufts Uni- versity School of Medicine; the Department of Pathology, Harvard Medical School, and Boston Hospital for Womea, Boston. R1:GULATORY CON7'ROLS OF ONCOTROPHOBLAST PROTEINS AND DI;VI:LOPMIiNTAL ALKALINE PtfOSPHATASES IN CANCER CELLS Regan isoenzymc (placental-typo alkaline phosphatase) and human chori- onic gonadotropin (1(CG) arc two oncotrophobiast proteins which arc onco- dcvclopmental gcnc products readily studied in human cancer patients and in several cxpcrimcntal systems. Three such mockls are: ( I) IieLa sublines TCRC-1 and TCRC-2, which produce Regan and non-Regan isoenzymes, (2) liep-2 and FL amnion cell lines acting as models for the reciprocal expression of dcvclopmcntal genea, and (3) in vivo modulation of developmental tcne ex- pression in licLa cclls. In this last instance, for examplc, Hel.a TCRC-l celts grow in immunosuppresscd rats, forming a tumor nodule which expresses a new oncoamnion (FL) isoenzyme, while the Regan isoenzyme disappears. Whcn thcse tumor cells arc returned to culture medium, however, the FL 8 9
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spodes disappean and the Repn isocnzyme reappears. This panicular modcl is ycpocted to pra,ve very uscful in intcrpretinj tbe various data obtained with oell cvItures and lurtson from catscer patients. A chronology of early devclop- mwt has already beca most helpful in tbis regard, since the counterparts of a tsumber of tumor ptotciru appaar as early as gametogcncsis and as late as ten .vects of nstation. FLshrrwn, W. H. and Singer, R. M. Cancer Rexarclr 36:4256-4261, 1976. OlJser aupporlt National Cancer Institute. From the Tufts Cancer Reuoarch Center, Tufts University School of Medicine. Boston, and the Depariment o( Auatomy, Fairkigh Dickinson University School o[ Dcatiatry, HackcnsacY. N. J. EVIDENCE FOR ALTERED GENE REGULATION iN HcLa CELLS RETURNED TO CULTURE AFTER GROWING IN IIrtJ.IUNOSUPPRESSED RATS A profound change of phersorypic exprcttan occurs when HcLa cells cloned for the exclusive production of the Rcgan isocnzyme of alkalinc phos- phataso am grown in immunosupptrsscd rats. In the course of a sequential al- tualioo in the isocnzynso axpt+wion during tumor growth, the Regan isoco- zymo disappears and an coca+moiao (FL) is+oeazymo becomes the dominant lorut. Tho presutt report dtscriba the isocnzymc regulation after these cells are tzturnod to culture for at least thrca months. These cells in culture dcmon- strato a dcasity-depcgsdcat alteratioa in isoenzyme profiles. The Regan iso- enzyme is ahe dominant isocnzynsa form in new, sparsely populated cultures. Whila the oocoauwion Loeazymo prcdomiaut.cs in the later hiyh-dcnsity stages of growth. Such a cell Iim which can be made to produce the Regan :soen- zytne by wlWting cclls In low density and which wiL retxpress the oncoam- nion (FL) isocstzymc at hith-ce11 density might be very useful in the study of isocazymo rc:ulation in human cancer cells. The eSects of prcdnisolone on tbcso cdls ia culture anc variable, depending on the time of its addition. A marked increase in cuzyme kvels is soen when the hormone is added early in tho growth ryck. Enzyme induction is restricted to the Regan iaocnzymc, while honswwo-mediated diminution in enzyme activity is confincd to the oncoamnion (FL) isocnzymc. Singer, R. M. (Fuiunan„ W. fl.) Cancer Rcsearch 36:4262-4265, 1976. OlJrer support: National Cancer Institutc. From the Department of Anstomy, Fairkigh Dickinson University School of Dcntistry, Hacktnsack, N. J. • IDENTIFICATION OF A IIUTANOL-EXTRACTABLE HUMAN PLACENTA-SPECIFIC ANTIGEN WITH ALKALINE PIfOSPHATASE ACTIVITY This study focuses on the placental rnembrane-associatcd antigcns «lcased by a-butanol, an organic solvcnt, which "ubilizes a number of cell membrane componentt. Most of the tissue lipid is extracted into the alcohoi traction kav- ing the glycoprotcins, polyuccharidcs and other water-solubk components in thc aqueous phase. In this series of experiments. bomolpenized whofe-term pla- ccntae from diffcrcnt individuals were treated with n-butanol and the immuno- gcns-containing aqueous fractions were used to raise bcterologous hyperimmunc scra in rabbits. Immunoclecarophoresis of the anti-placeota antiscra revealed at least siz antigcnic components in the placental extracts even after they had been completely absorbed with pooled mak serum prote'ms. However, the anti- scra so absorbed, designated (-PMS), still reacted strongly with extracta of normal adult intestinc and kidney. Similar reaction patterns were also obtained with individual butanol extracts of I1 various other rarmal adult tissues and with a composite extract containing equal amounts of each of the 13 individual extracts. Of the six antigcnic components in the placental extracts reacting with the (-PMS) antiscra, only the one associated with alkaline phosphatase activity retained its reactivity. Homogcnoous placental alkaline phoaphatasc confirmed this immunologic and enzynologic identity. Extracta from each of three pla- ccntae injected into three pairs of rabbits all produced an identical antibody reaction with the unique dcterminant(s) of placental alkaline phosphatasc. Ex- tracts of 14 other placentae reacted with each of these antisera to form a singk precipitin line of identity. Thcse results firmly establish that placental alkaline phosphatase is a characteristic placenta-specific fetal protein. Chang. C-H. and Angctlia, D. (Fisbman, W. N.) The Journal o/ Immunololy 117( I): 91-96, 1976. Other support; National Cancer Institute. From the Tufts Research Center and the Department of Pathology, Tufts Uni- vetsity School of Medicine, Boston. DIRECT IMMUNOPEROXIDASE STAINING FOR REGAN ISOENZYME OF ALKALINE PIIOSPIIATASE IN HUMAN TUMOR TISSUES Immunohistochcmical localization of Rcgan isoenzymc by a direct pcroxi- dasc-labcling technique was obscrvcd in certain ,ckcted cancer tissue cells by both light and electron microscopy. Among five turrwrs selectcd for cxamina- tuon, two cases (I 6c 2) that were histoclxmically confirmed by their estrcme sensitivity to L-phcnylalaninc (LPA) also ahowcd a spocific immunopcroxidase rcaction for Rcgan isocnzymc and did not show any difference between the histochcmical localization of Rcgan isocnzymc and the immunopcroxidase-stain- ing one at thc light microscopy kvcl. Thc Regan isocnzymc in these cancer cclls was scen prcdominantly in the cytopiasm containing diffuse and fine granu- lar «action producu, but it was also noted in the cell border area. Immuno- cytochcrnical studics at the electron microscopic level were confined to the tumor tissue from case I where Regan iaocnzymc could be specifically dcmon- stratcd on Ne plasma mcmbranc. On the other hand, cancer cells from case 3 10 11
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-17 oootainiai Lhornoar=inlno-.cnsiiivo alkaliao pbosphatase isocnzymc and from t.ases 4 and 5, ac3oGiod becauao ot tLa abscaco of alkaline phosphatasc activity, werc apparently untwctivo to the apcciAc unmunoperoxidasc staining. Based on thcao fiadlnaa, posoocidaae-labelcd antibody speciLc for human placenta has bceu suc,canfully used for dotcctin= Rogan Isoenzymc in cancer cclis, because ths identity ot piacental alkaline phosphatasc with Rcgan isocnzymc has been well establlsbod. Tha authors fcol that these LPA-acnsitive tumors rcprescnt a clao of caaccrs that sbouM be investigated from the point of view of cxpres- sion of embryoaic ycaca. ]•(iyayama, Ii., Doclltast, G. J., Memoli, V.. Gandbhir, L. and Fishmun. W. !f. Cancer 38(3):1237-11.46, 1976. OtJsar anpportr Nationaf Caocer Institute. From the Tufts Cancer Reseanch Ccnter and the Dcpartmcnt of Pathology. Tufts Univcrsity School of Mcdicinc. Boston; and the Departmcnt of Pathology. Poodvilk Hospital, Walpole, Mass. ~ TRACHEOBRONCHIAL EPIT}fELIAL MULTINUCLEATION, VIRAL INCLUSION BODIES AND MALIGNANT DISEASE This cpidemioio=iul study was undertaken to dctcrmine any possibk re- lationslups bctween lrschoobroochial cpithclial multinuclcation, viral inclusion bodia, and malignant disease. (II is already known that patients with malig- nant discase have aa increased incidctsce of multinuclcation and that thcre is a.casonal variatioa in the number of viral inclusion bodies in the ciliatcd epitbelium.) For this study, srncars from 4,150 patients - of whom 1.024 (yasp A) had various malignancies while the other 3.126 (groups B and C) were sulicrias from many types of nonmalignant condit'ans - were examined for the prcaeaoc ol viral iociusioos and multinuckated ciliatcd epithelial cells. Low dcyres of multiouclutlon were observed most frcqucntly in the summer in patknts both with and without known malignancy. Also, cytoplasmic inclu- siou bodies were seen least frequently in the summer and autumn in all patients. When group A and group C were oompuod. signifscantJy more patients with cancer bad viral luchraion bodies. The seasonal incidence of high percentagcs of awltiaucleatad colh in patkats with viral inclusions but without known cancer (gToup E) followed the Dusaal pattern of being lowest in the summer; but in tboso paticats with cancer (group D), perccntares of multinuckated cells were hijhe.r In the summu nwnths ihaa in any of the other scasona. These obscrva- tioos have led the authors to conclude that trachcobronchial cpichelial multi- nucleation seems to be tr{jyetsd (1) by malignant disease anywhere in the body and (2) by the acawnal prevalence of respiratory viruscs. Most important- ly, this work indicatos that respiratory viruscs may havc a apccillc effcct on the ciliatod cpithcGum of cancer paticnu. Cfsafow, J. st al. The Jownal of Rcprodt.ulvs Mcdiciuu 19(3) :141-146, 1977. From the Dcpanmeat ot Aoestlscsiolojy, New York University Medical Ccnter, Pfow Yort. 12 ENDOGENOUS GTYPE VIRUSES: DOUBLE AOENTS IN NATURAL LIFE PROCESSES About 20 d'Jfercnt vcrtebrato apoci•u arc now known to harbor C-typo RNA viruscs. In man, the presence o[ these particks is suggested by electron microscopic, biochemical and acro(ogic siudia, altDoujh a definite replicating human virus has yet to be isolatcd. Thcse cadojcnous viruses aro inbcritod through the germ cell and their production is regulated by tenetic information carricd in the hoat celL Two daascs of cndo^„-cnan C4ypo virvscs, ocotropic aad xeswtropic, which may have their counterp.rta in man, have been recognized in certain animals, particularly the ttaassc. Ecotsopic viruacs sprcad through the host, can be easily transmitted to cclla of ihe same species and can induce malignancy. Xcnotropic vinnes caanot infect cells from their host spocia but arc infectious for cells of hctcroloyotn apctics. The interaction between tbcso two types of viruses could result In the traasfcr among spccia of genetic in- formation rclating to normal life processcs and malignancy. The author coo- cludcs that cndogcnous C-type viruses may be positive rcgulaton of embryo- genesis, dillcrcntiation and normal cell development, as well as of the extrcascs of thcse processes, cell agioj, auroim.muoo diacase and cancer. Levy, J. A. BiomeJicinc 24(2):84-93, 1976. Other aupport: National Cancer Instituta. From the Department of Medicine and the Cancer Rescarch Institute, Univer- sity of California School of Medicine, San Francisco. ENDOGENOUS C-TYPE VIRUSES IN NORMAL AND "ABNORMAL" CELL DEVELOPMENT In this comprchcnsivic examinatioa of endo=enous C-type virusea in nor- mal and "abnormal" cell devclopmeat, the viruses' potential rofe in the proo- csses of normal maturation (embryo9eocsis aDd differentiation) aad aging (au- toimmunity and cancer) arc discussed. These C-type viruses, which are budding RNA viruses with lipoprotcin coats, art endogenous, /i.. inherited in the genes of many spccics, and have been implicated in cancer in aevcral speciq of animals. Study of the role of tlxsc viruses in nature has been aided greatly by the mouse rnodcl system, since mice develop diseases simifar to those of humans. Three classes of murinc C-lypc viruscs have been idcntiflcd by host range and xro- lugical studies. One of these classes, exemplified by the xcnotropic virus, is prescnt in early embryos, in cells undergoing normal differentiation and in cancer cells. The other clruses, ecouvpic and amphotropic, arc found primarily in animals with pathological cooditioos. All thcse C-typc viruscs are expressed spontaneously by cclls at certain frequencies and titans during the lifetimc of the mouse. A nonimmunoglobulin factor has becn identificd in normal mouso scra which specifically neutralius the xcnotropsc virus class of cadogcnous G typc viruses. Virus expression, thcrcfore, appears to be regulated by intracellu- lar as well as humoral factors. These observations suggest that C-type virusca play a role in natural processes, particularly normal maturation and aging. It 13
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is also susgutad that these muriae virwes may have counterparts in humans in whose tiusru eado8t:noto visira particks havts boen detectcd. Levy. J. d. Catscer Rsssarck 37 ( g):2957-29t18, 1977. OtJssr aapportt National Cancu' Institute. From the Cancu Rcseanch Instituta and the DcparUneot of Medicine, Univer- siry of Californla Modical Conter, San Fraacisco. ANfIOEN-SPECIFIC NONIMMUNOOIABULIN FACTOR TIiAT NEUTRALIZES XENOTROPIC VIRUS IS ASSOCIATED WIT/f MOUSE SERUM LIPOPROTEINS (MURINE C-TYPE VIRUS/ ULTRACEAfTRIFUaAL SF1'ARATION) Mouse acnt spoct>,cally neutralize xutotropie but not ecotropic viruses, this aeuttalizaelon being attributable to some soluble serum factor and not ctTccted by iatmunogiobtrliw. Because both senotropic virus and the solubk neutralizing factor (NF) are preseatio all house mice, the authors have suggested that NF plays a role in the regulation o( endogenous xepotropic viruscs. Intcraction of NF with the virus at the eell surface may change the internal mileu and thero- by a>Toct normal life prooeases. This serum factor is further characterized in this n:port. It is stable at pH 2-7 and resists ether extraction, freezing or brief heatirtg to f00' C. Ultratxntrifugal acparation of immunoglobulins and lipo- proteim dertwwastrates the association of NF with the serum lipoprotcins. Gel panseation chromato;raphy localius it in the region of lipoprotcins and IgM expiaiainj the previous eooclusion of others that it is an immutwglobulin. Vari- ous procedures aLo show Ihat It is twt the 70,000 molecular wcight glycopro- tcia(s) circulating in mouse acrutst in high titer. Ultracentrifugal fractionation of mouse lipoproteins indicates that most of the NF activity is concentrated in the deosity intervai 1.019-1.175 j/cros. Additional studies should determine whether NF is associatcd with one of the major classes of lipoprotcins or with a speciAc atiaor lipoprotein specics within this density interval. Preliminary evideaoo suggests that a pnotein moiety of NF may be important for neutraiiza- tioa. Low dernity lipoproteins in human serum have been reported to have a regulatory elYoci on ceU-nxdiated immunity, blocking thymus-derivcd (T) celi aetivatina and mixed lympbocyte reactions. The recognition, reported here, of a faUor associated with mouse serum fipoprotcins which spccitkally inactivata a virus represents a nooirrununoglobulin ncutraliwng rcaclion to an cndoa-cnoua virus by a boat. Loony J. C., Kane, J. P., Okszko, O. aat L.avy, J. A. Procecdinai o/ the Natlo+rof .tcadcmy o/ Sckncct o/ the Unitcd S/arca a/ .(rncr- k. 74:276-280, 1977. Other asrpportt National Canocr Itutitute and National Ifcart and Lung Irnlitute- From the Cancer Research Institute, the Cardiovascular Rcscarch )nstitutc and the Depanment of Medicine, University of California, San Francisco. INDUCTION BY CIGARETTE SMOKE OF ARYL HYDROCARBON HYDROXYLASE ACTiVITY IN TIiE RAT KIDNEY AND LUNG Previous invatigators have reported a relatioaship between the inducibiliry of aryl hydrocarbon hydroxylaso (AHH) activity and suaccptibility to the do- vciopmcnt of skin tumon in mice and pouibly lung cancer in man. This study of cigancltc-smoke-induood AHH aUivity represertts an attempt to understand the biochemical reasons for such a linkage. A IS-minutc exposure to a 1/15 dilution of cigarette smoke induced AHH activity in lungs and kidneys ftom Sprague-Dawky rau. In both lung and kidney thcre was a 2-bour lag bttwem smoke inhalation and the enzyme irsducilon onaot. Tborcafter, the AIiH activity increased very rapidly to peak about two hours later, reaching a maximal ao- tivity which corresporxf(d to three or four times that o( normal rats. Up to four successive inhalatiorn administered at 2-hour intervals induced both lung and kidney hydroxylase activities additively. Tbe maximal etIect corresponded to about 10 times the control value. Compared to the kidncy enzyme, the lung AHH activity was about three or four times more sensitive to small concentra- tions of cigarcttc smoke. Measurement of RNA and protein synthesis rr.quire- mcnu showed that protein synthesis was continuously required for the atab- lishment of the AHH induction but RNA synthesis was only necessary in the initial period of the phenomenon. To further characterize the lung and kidncy AHH activity, the half-life of the eigarette-smoke-induccd AHH activity was compared to mcthylcholanthrnnc-induced AHH activity. Ovcrall, the authors feel that their results support the hypothesis that the inducing agents in cisa- «lte smoke might not be polycyclic hydrocarborts. Van Cant(ort, J. and Gickn, 1. Inlernarlonal Journal of Cancer 19:538-545, 1977. Other support: Fond National dc la Recherche ScientJque. From the Laboratoire dc Chimie MEdicak, ltutitut de Pathologic, Li2ge, Bel- gium. ARYL-HYDROCARBON IIYDROXYLASE ACTIVITY IN LYMPHOCYTES FROM LUNG CANCER PATIENTS AND NORMAL CONTROLS The aryl-hydrocarbon hydroxylase (AHH) systcm, which metabolizes and is induced by a wide varicty of compounds, has been shown to bc genetically dctermined in certain strains of mice. It has also been suggested that the kvel of cncymc inducibility in human lymphocytes is gcrxtically regulated. In this study, a new Icchniquc involving radiomctric aasay was used to compare kvcls of AUIt activity in lymphocytcs of lung cancer patients and normal controls. Subjects included Il malc paticnts with histologically verified lung cancer (nine squamous cell carcinomas and two adertocarcinomas) arxf 1l age- and sex-matchcd controls. The lung cancer patients exhibited consi.krably more variation in AHH activity than the controls. In addition, the mcan AHH ac- tivity of the lung cancer patients was nearly four times that of the controls. Similar rrsults were obtaincd from comparisons made between the eight eiga- rctte sniokers in the control group and the agc-matched lung cancer patients. The cancer patients exhibited much more variability and a significantly higher 14 15
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N rtwa lcvel of AIiH tsctivity than the control group smokuz. While discussing tbo implieitiom of this work for lung cancer epidemiology, the authors suggest that AHH, by virttts of its possibk genetic polymorphic status couplcd with its ability to ttxtaboiiTe ptrUin chemical urcinogcns, may be of extreme value as a marker for dotecti4a of eancer suaceptibility. Guirgis, H. A., LyAcIY, N. T.. Mate, T., Hartis, R. E., Wclls, I., Caba, L., An- dcrsoq, J., Maloney, K., and Raakia, L Oncoloyy 33:105-109, 1976. OtJser asrpportr Natiooal Institutes of Health. From the Dcpartment of Preventive Medicirx, Crcighton Univcrsity, Omaha. ARYL HYDROCARBON HYDROXYLASE AC(7V1TY IN I'ULMONARY MACROPHAGES AND LYMPHOCYTES FROM LUNG CANCER AND NONCANCER PATIENTS. Previous rcporis have suggested that aryl hydrocarbon hydroxylasc (AHH) inducibility may be associated with the propensity to develop lung cancer. In order to study this hypothesized relationship, AHH activity was measured in pulmonary alveolar nuerophages (PAM) and peripheral blood lymphocytes from 47 paticnts with primary lung cancer and 56 paticnts bronchoscopcd for a variety of other medical probkms. In PAM from nonsmoking patients with- out eaaxr, the AHH level was 16s2 milliunits/ 106 cclls, whcreas nonsmokers with primary lung t:ancer had values of 24s3 milliunits/ 10' cells. Cigarette smolters eomistently had higher PAM enzyme values than twnsmokcn, al- though smokers without cancer and smokers with lung cancer had similar en- zyme ievela. Inductioa of AHH in eultured lymphocytes from nonsmokers was signiHcantly lower tor lung cancer patients than for rancancer patients. Enzyme induction was similar in lymphocytes from smokers whether or not they had lung canoer. Impottantly, with eells from individual patients without lung caaecr, a positivo correlation betwoen inducibility of AHH in cultured lympho- eyies and AHH activity in PAM was noted. On the other hand, with cclls front patients with primary lung eancer. PAM and lymphocyte vaiucs were not posi- tively oorrelated. Further investigations will be necessary to delineate the mecha- »istm rapoasible for the dissociation between AHII values in PAM and those in lymphocytes froca lung cancer patients. However, for whatever reasons this phenoenetson eaists, it may be of diagnostic value in the early detection of luns cancer. McLemore, T. L, Alartin, R. R.. Busbee, D. L, Richie, R. C., Springcr, R. R., Toppell, K. L., and Cantreil, E. T. CancerRcsearrh 37(4):1175-1181, 1977. Ot/ser aupport: National Institutes of Health, National Cancer lnstitutc and the American Canccr Society. From the Department of Medicioe, Baylor Collegc of Medicine, Houston. and Departments of Biological SCKncYa and Basic Health Scicncn, North Tcaas State University and the Texas College of Ostcopathic Medicine, Denton. IN VITRO R:I)UCTIOti OF ARYL HYDROCARBON HYDROXYI.ASE IN HUMAN PULMONARY ALVEOLAA MACROPHAGES BY BENZANTIiRACENE The polycyclic hydrocarbon, beaxanthtsceno (BA), is capable of inducing aryl hydrocarbon hydroxylaso (AHH) in human pulmonary alveolar macro- phages (PAMs). In the study presented here, time and dosarcaporue curves for maximum in v(rro induction of tho AI{H enzyme system in PAMs were established. In addition, a positive correlation was demonstrated between the level of Altli activity in PAMs frulily lavaged from the lung and tLe degree of induction of AHf( in cultured PAMs from individual snwkers and non- smokers. When PAMs from either smoken or nonsmokers were cultured for 24 hours in the presencY of 10 µM of BA ptr vial, significant induction of Alili occurrcd. Induced values of AIiH were over thrcc times those of noo- induccd valucs for PAMs from sunokers, while induced AHH values in PAMs from twnsmokcrs wcrc over four tima greater than the noninduced values for thc saox individuals, indicating that BA is a good inducer of the AHH system in cultured PAMs. Measurements such as these of the inducibiliry of AHII in cultured human PAMs could provide an experimental system suitable for study- ing the mechanisms responsible for the initiation of pulmonary earcinogcnesis. McLcmore, T. L. and Martirt, R. R. Canccr Lctlcrs 2:327-333, 1977. Other aupport: National Institutes of Health. From the Dcpartmcnts of Medicine, and Microbiology and Immunology, Baylor College of Medicine. Houston. RELATIONSIiIP BETWEEN LEVELS OF ARYL HYDROCARBON HYDROXYLASE AC17V1TY AND SUSCEPTIBILTiY TO 3-MLiTHYLCHOLANTHRENE AND BENZO(a)PYRENF.INDUCED CANCERS IN INBRED STRAINS OF MICE Susceptibility to 3-mcthykholanthrene (MCA)- and benzo(a)pyrcrx (BP)- induccd,fibrosarcomas (and carcinomas) is linked to naturally occurring differ- enccs in aryl hydrocarbon hydroxylase (AHH) kvels, that is, where AHII in- ductibility segregates as a codominant gene, or where noninducibility is domi- nant. Moreover, under conditions where AHH levels arc artificially altercd, as occurs aftcr treatment with the chemical defoliant 2,3,7,8-tetrachlorodibenxofaJ dioxin (TCDD), the inherent resistance of AI1H nonrrsponsive mice to MCA- induced tumors can be overcome. Mouse genetic model systems show that when Alllf responsivcncss to MCA treatment segregates as a single autoeomal dominant gcne, eitlxr a single autosomal codominant gene or noninducibility can be dominant depending on the strains of mice employed. In each of the three genetic regulatory systcros described here, the kvel of AIfH irnducibility is corrclatcJ (or linked) to susceplibility to'tumor induction by MCA. Where AHIi inducibility segregates as a single dominant gcne, susceptibility to MCA- induced fihrosarcoma is correlated with expression of that gcne function. Susccptibility to BP also correlates with high levels of AHII inducibility. High doses of TCDD given.48 hours prior to MCA result in tumor incidences simi- 16 17

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