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Council for Tobacco Research

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

Date: 30 Jun 1971
Length: 49 pages
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ANNUAL REPORT
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SCIENTIFIC ADVISORY BOARD to The Council for Tobacco Research-U.S.A., Inc. as of June 30, 1971 SHELDON C. SOMMERS, M.D., Chairman Research Director, The Council for Tobacco Rcscarch-U.S.A., Inc. Director of Laboratories, Lenox Hill Hospital Clinical Professor of Pathology Columbia University Collcge of Physicians and Surgcons New York, New York HOWARD B. ANDERVONT, Sc.D. Scientific Editor (retired), The Journal of the National Canccr Institute Bethcsda, Maryland RICHARD M. BING, M.D. Director of Cardiology and Intramural Medicine Huntington Memorial Hospital, Pasadena, California Professor of Medicine University of Southern California School of Medicine Los Angeles, California McKEEN CATTELL, PH.D., M.D. Professor Emeritus of Pharmacology Cornell University Medical College, New York, Ncw York ROBERT J. HUEBNER, M.D. Chief. Viral Cardnogenesis Branch National Cancer Institute Bethesda, Maryland LEON O. JACOBSON, M.D. Dean of the Divuion of Biological Sciences Regenstein Professor of Biological Sciences Univcraity of Chicago, Illinois CLARENCE COOK LITTLE, Sc.D., LL.D.. Ltrr.D. Scientific Director, The Council for Tobacco Rcsearch-U.S.A., Inc. Director Emeritus, Roscoe B. Jackson Memorial Laboratory Bar Harbor, Maine CLAYTON G. LOOSLI, Ptt.D., M.D. 1/astings Professor of Medicine and Pathology University of Southern California School of Medicine Los Angela, California KENNETH MERRILL LYNCH, M.D., Sc.D., LL.D. Chancellor and Professor Emeritus of Pathology Medical Colleae of South Carolina, Charleston, South Carolina WILLIAM F. RIENHOFF, Jtt., M.D. Professor Errreritus of Surgery Johns Hopkins University School of Medicine, Baltimore, Maryland ROBERT C. HOCKETT, Ptt.D. Acting Scientific Director J. MORRISON BRADY, M.D. JOHN H. KREISHER, Ptt.D. Assaciate Scientific Director Associate Scienuf;c Dirrctor VINCENT F. LISANTI, D.M.D. Scientific Associate CLARENCE COOK LITTLE, Sc.D., LL.D., Litt. D. 1II88-1971 Dr. Clarence Cook Little, internationally known geneticist and cancer «searcher, dicd December 22, 1971. He had been Scientific Director of The Council for Tobacco Researeh-U.S.A., Inc. and its predecessor since shortly after the latter's establishment in 1954. I(c also served (or many years as Chairman of the Scientific Ad- visory Board, an elective position for which only Board members votc. Former president of both the University of Maine (1922- 1925) and thc University of Michigan (1926-1929), "Pete" or "Prczy," as he was known to colleagues and itudcnts, was a tower- ing figurc as a man and as a scientist. Warm, dynamic, an engaging personality with a sharp wit, Dr. Little was one of the first scientists to discover evidencc ol a viral agent in cancer, an observation that led eventually to the generalized virus theory of cancer. He also did pioneering work in immunology and was one of the early re- searchers into the genetic effects of radiation. Dr. Little founded the world-famed Jackson Laboratory in Bar Harbor, Maine, in 1929. He served as its director until 1956 and as director emeritus until his death. He formerly was Managing Director (1929-1945) of the American Association for the Control of Cancer, which is now the American Cancer Society, and was a Fellow of the Na- tional Academy of Sciences and a member of nurnerous other scientific societies. He authored many books and articles and his 1939 book, "Civilization Againsl Cancer," is still considered a classic for the layman. For 17 years Dr. Little gave The Council inspired scientific leadership and guidance. His contributions were many and important. He will be deeply missed.
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CONTENTS Introduction 5 The Cancer Program . . . . . . . . . . . . . . . 6 Cardiovascular Research . . . . . . . . . . . . . . 1 1 Chronic Rcspiratory Discascs .. . . . . . . . . . . . 14 Ncurophysiology, Psychophysiology and Pharmacology ..... 16 Other Studics . . . . . . . . . . . . . . . . . 18 Abstracts of Reports . . . . . . . . . . . . . . . . 20 Psychophysiological Studics . . . . . . . . . . • 20 Carcinogcncsis Studies . . . . . . . . . . . . . 23 Cardiovascular System . . . . . . . . . . . . . 34 Respiratory Systcm . . . . . . . . . . . . . . 42 Tissue and Organ Culture . . . . . . . . . . . . 55 Studies at the Cellular Level . . . . . . . . . . 56 Pharmacology and Psychopharmacology . . . . . . . 59 Metabolic Studies . . . . . . . . . . . . . . 69 Chemistry and Biochemistry . . . . . . . . . . . 71 Epidemiology . . . . . . . . . . . . . . . 76 Active Pro jccts . . . . . . . . . . . . . . . . . 79 Completed Projects . . . . . . . . . . . . • . . . 89 I I Introduction Previous Annual Reports have stated the general raearch policies of The Council for Tobacco Rescarch-U.S.A., Inc. as conceived by the Scientific Advisory Board. These policies, which continue to guide the program, emphasize study of the etiology of those diseases that are reputed to be related statistically in their incidence to cigarette smoking. Such diseases, especially cancer, cardiovascular ailments and chronirc respiratory afflictions, arc not only leading causes of tnorbibity and death since the conquest of major infectious diseases but are universally rowg- nizcd to be of multifactorial ocigin and to be strongJy influenced by con- genital predispositions. The role of tobacco use in their etiology, if such exists, can therefore be defined or measured only within the context of a growing comprehension of the total etiological picture, involving knowledge of the complex interactions between endogenous and exogenous factors. As new concepts develop with respect to etiological mechanisms that may be involved in the genesis of these constitutional diseascs, these con- ccpts in turn suggcst new biological test systems for determining whether chronic exposure to cigarette smoke or some ingredient thereof can con- tribute to the operation of these mechanisms. In this way it is to be hoped that a series of progressively more meaningful test systems will evolve as ctiological understanding advances, to supersede the crude and inadequate mcthods of the past. All investigators in the field are well aware that scientific progress takes place "in slow motion." The description of progress in a single year is akin to attempting to depict the progress of a baseball gaine by desaib- ing one inning. Moreover, since the progress of studies in one single pro- gram such as that of The Council is interrelated in a very complex manner with the progress of many other studies throughout the scientific world, the report of any one program can hardly hope to present anything resembling an integrated picture of the status of knowledge In any particular beld or arca of medical science. Recognizing these limitatiam, we have attempted only to mention in this report a few selected findings that have emerged in published form in the year ended June 1971. These aclectiorts do not in any way minimize the potential importance of many contributions by other investigators that do not receive particular mention here. S1nae all these contributions have entered the "public domain" of science, they will be assimilated into the body of total knowledge and comprehension that occurs through the process of "digestion" and anabolism by the great eoafratcrniry of investi- gators in the many disciplines involved. Roat:ar C. Hocuerr Acting Scientific Director 5 C~ H U .y.. :~... e
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w The Cancer Program Viruses and Cancer Viral aspects of cancer, including the human disease, are being in- tcnsivcly studied in many laboratories throughout the world. Reports of advances in this active field seem to appear almost wcckly as scicntists strive to pin down exact evidence that human cancers, or at lcast some of them, are caused by viral factors. Perhaps of greatest prominence in this field is the general viral gcnomc concept. This postulates that a C-type RNA viral genome exists in all mammalian cclls but remains "switched off" until, for some rcason or reasons still largcly unknown, it becomes activated or "derepressed" and triggers the development of cancer in forms dctermined by genetic programming of the strain or species. Deoelopmenl of a Tesl System A Council-sponsored study has been aimed at development of super- sensitive, virus-primcd tissue-culture test systems for measuring the relative potentials of various clxmiuls, in collaboration with the virus, to produce "malignant transformation" of cells. For example, rat embryo cells infected with Rauschcr leukemia virus have been trtated with various doses of 7/12-Dimcthylbcnz(a)-anthraccnc (DMBA). Morphologic transforma- tions indicative of neoplasia were san. On the other hand, there was no transformation in uninfocted rat embryo cells similarly treated or in infected cells rot treated with tho DMBA. When transplanted into newborn rats, the transformed cells produced subcutaneous sarcomas, but no turnors resulted from like treatment with the infected or DMBA-treatcd, untransformed Cells. The work showed that itt this rtsodd, at leasi, both chemical and vinu wert necessary fa cdi tramdotmition. According to the investigator, the findings suggest that tbe C-type RNA viral Dcaome of the Rauschcr lcu- kemia virus provided specific oacoSene information for the malignant transformation. I It must tx rccogniud that this model doea not duplicate real life situations. Rather it exaggerates certain factors and eliminatcs others for the sakc of supcrscnsitivity. The effects of chemiuls on cell transformation arc greatly enhanced by priming with virus, while the protective effects of the in- tact animal's immunity system are eliminated. Hence the model can be ex- pectcd to show positive effects by chemical agents which, on the basis of earlicr in vivo test systems, have been considered totally inactive as "carcinogens." Such indications from the new supersensitive systems therefore do not simply or automatically imply that such substances are either "dangerous" or "safe." The model is only one new tool that provides comparative ex- perimental values on a single scale. From many such evaluations a relative "activity scale" or set of scales may develop that will permit more direct and morc ncarly quantitative comparisons of very fcebly active compounds and mixtures with those of high activity. Any attempt to extrapolate the findings from such a model to real life situations, especially the human, will require even greater care and discretion than heretofore. A hopeful potential for the model lies in the ultimate possibility of including human tissue cultures in the system for comparison and contrast with those from other species, under defined, controllcd and closely anal- agous conditions. A rnorc distant goal is development of a method (or assessing the relative degrees of cancer suseeptibility in human patients. Another virus-relatcd study of The Council, completed during the year, sought to determine whether a relationship could exist between the occurence of cancers in humans and the presence of cancer virus in house- hold animals such as cats and dogs. The work concluded that no such relationship exists. 'ihe scientist who carried out the study reported that whilc C-typc viruses in an active state have been found in the domcstic cat, thcrc is no evidence that they spread to dogs or to man or even from cat to cat. It appean, under natural conditions, that feline C-type viruses are inherited as indigenous latent genes which may activate in the fetus or later in life. Thc investigator cautioned that susceptibility of certain human cells to in vitro infection with these C-type viruses does not imply infection in the living organism. The samc scientist, reporting on another facet of the study, disclosed indications that fclinc sarcoma virus can transform human osteosarcoma cells. 71tis example of cell trarnfoematlon by a non-host virus seems of cornidcrable signifkaaoe in rirtsoprear >F F Ilk : Findings aainod by..tfilr hiratlStlot yj Ilk 6nndl-supportcd project were applied to later nactrch, lunded by the Nulonal Cancer Institute, in which human cancer cells were Injoctod into Ifie fctusa of pregnant cats. Some of the cat fetusrs developed tumors of human cells in which wu,e ~ ~ t n ,-• 4....t 6 7
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found particlcs similar to the known C-type eancer virus particles of several animal species. Praent immunological evidence suggests that thcse may be the long sought human cancer virus particles. If this finding is eonfirmed, it should grcaUy facilitate testing in man the many implications of the viral gcnomc concept of cancer etiology that appear to present a rathcr wcll- intcgratcd picture in several animals. It might also makc possible the addi- tion of virus-primed human cell cultura to the in vitro test systcm that has been described. Ot1ur Virr.s-f.ancer Slruliea Lymphosarcoma in rabbits, a rare tumor fn that particular animal, has been the subject of another Council-sponsored project. A researcher studicd one strain of rabbits among which a number had dcvcloped lympho- sarcomas. He analyzed the pedigrea of the affected animals and found that inhcritancc indicated an auto.omal recessive gene conferring susecpti- bility to lymphosareoma. The findings, the investigator noted, were con- ceptually compatible both with genetic susceptibility to malignant lymphoma and with vertical transmission of a virus. The neoplastic involvement of various organs in the rabbits rescmblcd in many ways visceral lymphosaroomatosis of cats which has been proved to be caused by fcline leukemia virus. According to the investigator, his findings on lymphosarooma and its hereditary basis provide a new and important model for studics of the pathogenais of neoplasia. Still othcr studies being supported by 'Iln Council in the virus arca include: oneogeny and the antiviral action of interferon, effects of respira- tory and oncogatic viruses In organ cultures of human respiratory tract tiuuc, and mechanisms of suppression of cellular immunity by carcinogenic hydrbcarbons. Srwoke Exposure Devkea In its search for more realistic bioassay systems, The Council has (or many years supported studia involving cigarette smoke inhalation by animals. A major aspect of this undertaking is, of course, the design and production of a "smoke exposure device" that permits exposure of cxperimcntal animals to smoke inhalation in a manncr and updcr condi- tions that resemble human smoking as closely as possible. Previous Annual Reports ot'Rte Council have outlined.everal strict criteria for the functiop of such devices and these have subsequently been extended. The Council has developed a machine which has been undergoing tests in actual projects. While !t has already demonstrated its value in several researches, a number of small but important improvements are being made to refine and define its operational characteristics. In using such a device, perhaps the most difficult problem is to measure how much of the particulate phase of the amoke actually reaches the lung surfaccs of animals that normally breathe through their noses. Such mcasurements arc being carried out with tagged smokes. The machine now being usc-tated by several Council grantcts has provision for inscrtion of a Cambridge filter that tr.rnovcs particulate material and thus permits comparison of the effects of whole smoke inhala- tion with those of inhaling the gas-vapor phase alone. Ilandlina and Standardiaation oJ Anirxeli It is important to bear in mind that handling animals and confining them in a smoke exposure device is very stressful to them. Hcnce "machine controls" that are subjected to the same handling and confinement in a functioning device, but without actual smoke exposure, must be included in experiments as well as unhandlod "cage controls." Comparisons of thae two acts of controls can be expectcd to disclose some of the biological cffects of these particular stresses. Thcac, at kast, can then be taken into account in the interpretation of smoke inhalation experiments in which handling and confinement stresses cannot be sep.rated directly from smoke effects. Significant biological effecta of atras, otiginany observed in such "machinc eontrols," have now become a subject o( invatigation in their own right as part of The Council's pro=rua. In connection with artwke inhalation as weA as other kinda of studia, The Council has sought improvement in the staadardizatbn of anlmah. Animals used in most Council-suppotted ttacarrh are delivered by oaarcart section and arc substantialy free from pstbogens. Alro, their virus pro6la arc determined as fully as pottsible as well as the atatut of the C-type RNA viral genome expr>~uion. Many eanonc researcbers use inbred strains ol mice because of the great variety o( strains with known congenital tumor susceptibilitia. However, because there sometimes arc advantagu in using hamstcn, which have larEer lungs and greater blood volume, studies have been started with this species. Some inbred hamsters strains am now available. 8 9
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Exposure oJ Cultures to Sr..okt In onc smoke study, three types of primary cultures from Sncll micc- kidney tissue, embryonic lung organ and lung explants-and one established cell line from Swiss mice were used to asuy the biological cQccts, in virro, of the gas phase from charcoal-filtered fresh cigarette smokc. Rcpcatcd exposure to puffs of the smoke did not produce any significant changes in thc Snell mouse cultures. The exposed Swiss mouse cells showed an cn- hancemeat of growth, mitotic index, and cellular atypism. No morphological ccll transformation was observed in any of the exposed cultures. Such in vitro studies arc mainly useful as aids to the interprctation of whole animal studies. Being by necessity "unphys'alogical;" they cannot bc extrapolated easily or directly to normal life conditions. The researcher who performed this study under a Council grant had previously reported that Sncll mice, with viral genomc expressed, exposcd chronically to inhalation of either whole smoke or its gas-vapor phase (devoid of the particulates or "tars") developed lung adcnomas and adenocarcinomas of the same kind that they develop spontaneously. None of the animals developed squamous cell lung tumors. Mice of another strain, lacking viral gcnomc expression, showed a dramatically lowcr incidence of adenoma and adenoearcinoma under the same conditions. In a study of the incidence and appearance of lung tumors and pneumonitis in mice, a Council-supported scientist immunized mice with an influenza virus and then exposed them and control mice to sub-Icthal and highly lethal nebulized clouds of fresh virus. The controls succumbcd quickly to the lethal cloud of virus and had extensive and persisting pulmonary changes following exposure to sub•kthal clouds. The immun- izcd mice became only mildly ill and survived the lethal clouds, though large amounts of virus were found in their lungs, which soon returned to nornial. The researcher noted that immunization, while not preventing infection, was beneficial to the animals exposod to airborne influenza virus. It is known that many mouse stnins develop adcnomas in the bron- chiolo-alveolar regions «gardlcu of what is, or is not, done to them. Adenomas do not seem to arise in the large bronchi or in the trachcaa of thcse animals. Is there a difference in susceptibility of these tissucs to chemical agents? Or is it that these inciting agents, when used cxperi- mentally, do not reach these different tissues in equal amounts? In an effort to learn some answers, The Council has supported work in which fragments of tissue from lungs, large bronchi and trachcas, rc- spectivcly, of BALB/e/Cb/Se mice were impregnated with methylcho- lanthrcnc (MC) alone or mixed with tak powder in concentrations of 1:10 i and 1:100. Thc fragments were then transplantod into the subcutaneous tat pads of syngencic hosts. Results indicaicd that the sensitivity of bronchiolo-alvcolsr cpithclia to MC was higher than that of trachoo-btonchial epithelia. However, the lattcr were not wholly resistant to the MC since there was almost constant tumor growth after impregnation with the carcinogen. Thus, there does appear to be some difference in tissue susceptibility. A similar experiment with subcutaneous grafting of tracheal, laryngeal, bronchial and lung tissues, followed by a single intraperitoncal injection of urcthane, produced adenomas only in the grafted lung tfuue, showing a tissue specific cffcct of this particular agent and suggesting that the agent was transportcd by systemic routes to the locus of action. Other Cancer Studies In other work related to cancer, The Council is continuing support of cfiorts to develop "positive" models for the production of squamous cell lung cancer in animals. Guide lines arc few In this area, but i( sonx simplc, easily reproducibic method could be devised, it would be very helpful in many research areas. Il lung cancer of the type reported preva- lcnt in man could be produced repetitively, at perhaps the 20 perrsat kvel, in an easily handled animal, it would then be possible to undertake studies to sce whether many facton-age, sex, horrnoncs, irritants, diet, stress, infcction, and othcrs - mightin8uence the level and rate of incidence of thc tumor. Also being continued is a project in which a concentrated stream of fresh cigare(tc smoke is being blown directly on the skins of mice of a strain previously used in a skin-painting test with tobaceo-smoke eon- dcnsatc. Among other ongoing projects being supported are: hctcro-trans- plantation studies with human lung uneer, the pathogenesis of urethane- induced lung adcnomas in mice, and host factors in lung cancer and other lung ailments. Cardiovascular Research Ensyme Related to .ltherosclerosiu A study involving an enzyme has turned up findings that may be significant in regard to atherosclerosis. The Council-supported project used 10 11
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scgmcnts of human iliac arterica incubated with human serum. Samples were hcated to inactivate the enzyme Iccithin-cholcstcrol-acyl-transferaae (LCAT). It was found that when LCAT enzyme was present in the ineubation medium, virtually only free cholesterol left the arterial wall. However, whcn LCAT enzyme was absent, only cholesterol ester lcft the wall. The results suggest that a decrcase in setvm free cholcstcrol, caused by a serum cholesterol csterifying enzymc, possibly LCAT, might bc important in promoting the removal of free cholcstcrol from the artcrial wall. According to the investigators who performed the work, thc results arc consistent with findings that there is a tendency for LCAT activity in the serum to be higher in persons with hypercholcstcrolcmia greater than 300 mg/l0l) ml, which may reflect a mechanism for protecting the arterial wall from excessive frce cholcsterol. The scientists said their findings may be particularly important because they have found that cholcsterol estcrification is significantly impaired in persons with acute myocardial infarction and chronic coronary artery disease when compared with age-matched controls. Thus, they said, a deficiency of cholesterol esterifying enryme, possibly LCAT, may be important in the etiology or rate of development of atherosclerosis. Coronary Mi:crocirculation Continuing his work on total and nutritional coronary flow, another grantee has reported a new technique for determining the velocity of red cells in the capillarics. He has described capillary blood flow as that por- tion of the total coronary flow involved in the nutritional function of heart muscle, while total coronary flow represents all the blood entering the heart circulation. . To measure the velocity of capillary red cells, the investigator dcvel- opcd a method showing the framc-to-frame progrcas of individual red cclls in moving pictures of the capillaries. The technique also permits measurc- ment of the diameter of eapillaria as it changes from framc to frame. Data from one part of the project indicated that norcpincphrine re- sulted in opening of capillaries, that nitroglycerine slowed the flow in the capillaries, and that methachoilne resulted in the complete arrest and even retrograde flow. While intcrpretation of these findings will depend on con- tinuing study, the researcher believes the work will provide new information about the coronary microculatlon. Another aspect of the sttady was a further elaboration of previously reported findings on the measurematt of total and nutritional coronary flow by means of a coincidence counting system using rubidium-84. Working C" with this system, the grantee found that norepineprine caused a dispropor- tT tionatcly greater increase in total as compared with efiective flow, and the lf: same result also occurrcd with nicotine. Isoprotercnol, on the other hand, led to a proportionate increase in both nutritional and effective coronary flows. In another Council-supported study, mongrel dogs were exposed to cigarette smoke inhalation and also to injection of various pharmacologic agents. A reduction in aortic blood pressure during smoking by the anes- thctizcd animals was seen. This was attributed in part to the effcets of nicotine on the central nervous system and in part to vasodilation caused by the action of released catccholamincs on certain receptors in the peri- phcral vascular system. Kinina and 1nflammatory Reactions Another investigator has been studying kinins, polypcptidcs that are involved in inflammatory reactions. There is evidence of vascular permea- bility occurring independently of known mediation systems, and the kinin- forming system is being investigated as a possible humoral mechanism in the induction of vascular injury. Physical properties of the components of the kinin-forming system fn rabbits arc being studied. The characteristics of human components are comparable. It is hoped that this work will indicate the substance or sub- stances that may affect inflammatory reactions in the lung, particularly in hypersensitivity states. Framin6ham Ileart Study The Council has made a grant related to the Framingham Heart Study, effective July l, 1971. This study was to have been ended for lack of federal funding, but its long-time director and several associates sought and obtaincd outsidc support to keep it going. The Council's grant is for an investigation of smoking elassca, risk faetors and cardiovascular disease in thc population being followed by the project. Other studies being supported include: nicotine and various aspects of cardiovascular function, carboxyhcmoglobin and cardio-respiratory function, smoking and air pollution in human myocardial metabolism, in- flucnce of smoking and nicotine on lower limb circulation, sensitivity of vascular tissue to nicotine, and the effect of smoking on regional cerebral blood flow in smokers and nonsmokers. ' L>i . ~ F-- C.~ 12 13
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Chronic Respiratory Diseases Hyaline Membrane Disease A significant finding in regard to hyalinc membrane discasc (I1MD) has come from a Council-supported project that encompassed the analysis of 387 autopsies on newborn and stilaborn infants. It was found that the adrenal glands were 19 percent lighter in infants with HMD than in those free of the disease. Those without the disease had a grcatcr number of adrenal cortical cells. Also, a positive eorrrlation was found betwecn the presence of infeetion arising before birth and the absence of HMD. In connection with surfactant, which appears to play an ilnportant role in this disease, it is interesting that anencephalic neonates who had little or no adrenal fetat cortical zones and half-sized adult zones, were found to have only 45 percent the mass of onmiophilic granules in pul- monary type 11 alveolar cells found in non-ancnccphalic control infants. These granules reportedly arc the anatomic representation of surfactant. The investigators who conducted the study noted that the corticosteroid mechanism "oficrs the attractive possibility of relatively simple prophylaxis against hyalinc membrane disease in certain high risk gestations." But, they caution, "This temptation to treat should be strongly resisted until the mechanisms in question have been explored in the good experimental modcls of hyalinc membrane disease now available. The need for such. caution is also supported by the observation that the doses of cortico- steroids and ACTH used to increase surfactant levels in the lungs of fetal lambs have induced toxic changes in other organs." .! ppqlaclsian Coal Minera A quantitative morphologic study was undcrtaken of lung and hcart structurc in 322 Appalachian miners who were classified by age, duration ot mining exposure, rank of coal mined and smoking habits. Right vcntric- ular hypcrtrophy as evidcnce of cor pulanonale was common aftcr age 50. The volume of macular dust lesions, induding associated silica crystals, fibrosis and focal emphysema, increased with age but these dust lesions were not deemed to be solely rrsponsible for dyspnca since mincrs with dyspnca also had a generalized form of emphysema. Smoking was associated with a modest increase in broochitis and emphysema in bituminous mincrs but showed little or no rrlation to the extent of such lung conditions in anthra- cite workers, and nonsmoking tninus also developed these disorders. , Above-ground air pollution may play a role since the wives of some bi- tuminous miners seem to be nearly as much affected as their husbands by cough, phlcgm, wheezing and breathlessness. The association of pncu- moconiosis with coal rank was found to be fortuitous. Microcirculation of the Human Lun6 During the year a Council grantee published a report dcscribing the blood and lymphatic microcirculation of thc human lung that provides a clear, concise summation of major findings in this particular field. The investigator, long involved in lung research, gave the results of corrclatod macroscopic and microscopic studics of the lungs of humans, beagle dogs and mongrcl dogs. The report was primarily concerned with angio-archi- tectural relationships between the bronchial and pulmonary vascular sys- tcros, and the author noted that the relationships arc distinct in each. The bronchial arteries arc directly connected to each other, establishing a "parallel" vascular supply. The bronchial veins also form the peribronchial vcnous plexus. Howcver, the pulmonary eireulation, the pulmonary arteria and veins form terminal, intralobular branches. The bronchial and pulmonary artcries also communicate in the bronchioles through a common capillary network. The grantee reported he has been unable to observe broncho-pul- monary artcrioarterial or artcriovenous anastomoses in entirely normal adult lungs of beagles, but said they seem to be normally present in the lung of the human fetus and infant. He said they probably vanish in child- hood but reappear in the adult lung with the onset of even minimal pul- monary inflammatory rcactions or procaacs. The acicntist also published anotber report in which he described oells with the ultrastructural characteristics of neurosectetory cells in the lung lobuics of prematurely born infants. He had previously suggested tlhe existence of these cells on the basis of light and Auotrsocnce microscopy and noted that analogous cella had bcen reported by others. He said the possibility exists that thex cells may be involved in the regulation ot lobular growth and in modulation oi normal periaatal cardiopulmonary adaptation, and that they may play a role in respiratory d'utress syndrome. Pul monary .!l eeolar Macroplsaaea What is thc origin of the pulmonary alveolar macrophages? There has been considerable controversy as to the cellular precursors of ttxse cells, which comprise a major defense mechanism of the lung. A Council-sup- I 14 15
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ported study has elicited evideoce to indicate that at lcast some of the macrophaga originate from bone marrow. The study used gcnetically related mice and substrains to invcstigate the possibility that macrophagcs originate in marrow atcm cclls. Succcss of the expcrirnent rested on the fact that the gcnctically rc- lated mice carry a nonspecific, readily identiftable prc-albumin cstcrasc marker. Lethal x-irradiation and injections of bone marrow from the mice 1ed to findings confirming that at least some macrophagcs originate in bone marrow. This particular work was part of a project that is moving into the biochemistry of macrophagcs. Another Council grant.te provided a descriptive report of the appear- ance under the electron microscope of alveolar macrophages obtained by endobronchial lavagc from smokers and nonsmokers. He said that in general maaophages from smokers contained larger and more heteroEcrxously structured inclusions than those from non- smokers. Howcvcr, there was considerable variation in the number, sizc, and hctcrogcnicity of inclusions within individual macrophagcs from smokers, suggesting that various staga of phagocytosis were represented in lavage samples. Further, the researcher said, some substanccs that arc phagocytizcd but not digested probably accumulated in thc larger cetero- gcncous inclusions of smokers. Other studies of lung macrophaga have shown that a greater number arc recovered by lavagc from smokers than from nonsmokers, and that the macrophagcs from smokers show unimpaired phagocytizing capacity in vitro. Neurophysiology, Psychophysiology, and Pharmacology Nicotine and Memory Consolidation For several years The Coundl has been supporting research into the cftcets and action of niootine on memory consolidation. One researcher has found that rats put through a training regime (maze solving and lever pressing) karn at a measurable rate. Howcvrr, if the animals are given an ~ ~ elcctric shock' after a training session, they tend to "unlearn" everything ~ thcy had previously acquired. Apparently the shock caused an amnesiac condition and prcvcntcd consolidation of the learning process. Whcn the animals were treated with nicotine prior to a training scssion, it was found that the subsequent electric shock did not affect mcniory consolidation; the animals largely retained what they had lcarned, apparcntly because the nicotine blocked any efiect of the shock. The time of nicotine treatment was important in this experimental procedure, the effect being maximal some time after administration. Cotinine, a nicotine mctabolite, also was effective immediately, which suggcsts that the lattcr substance may be the active agent. Other parts of the project included tcsting the rolc of the scrotonin precursor, 5-hydroxytryptophan; age difier- cnccs of the animals; differential housing; reduced brain amine levels; and aminc changes at the cellular level. In work in a related area, a Council-supported researcher found that chronic treatment with nicotine induced an increased turnover rate and utilization of noradrcnalinc in thc brains of rats. Pretreatment with nicotine did not affcct ttic action of sedative, antidepressant and central stimulant drugs in the animals' brains. In short, the chronic administration of nicotine appeared to stimulate the brains of the rats, making the animals function more effectively. Sex /lorrrsonea and Emphysema A potentially significant report during the year came from a Council grantcc who has been working on the experimental induction of emphysema in rats. He has sought to determine whether such induced disease would lead to pulmonary hypertension and cor pulmonale as in man. It was found that daily injections of progeatcrone (the female sex hormone) and mcdroxyprogcstcrone (in a dose 1/50th that of progcsteronc) prevented experimental induction of emphysema in the rats by a method that other- wisc was successful. Another grantee, treating rats daily with nlcotine injections, found an elevation in urinary excretion of eatecholamina that peaked after three days. After seven days of continuous trutment, there was a marked fall-off in the urinary levels, and by 14 days the catecholaminc excretion was essentially normal. This suggests a physiobgieal accommodation or adjust- mcnt over a period of time to chronic administntion of nicotine. The scientist explored three possible mcchanisms that might be rapon- siblc for the return to normal of the elevatcd urinary catechulamincs. He concluded that tolerance to the nicotine-induced elevations of urinary 16 17

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