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.k SCIF.NTIFIC ADVISORY BOARD to The Council for Tobacco Research-11.S.A., Inc. as of June )U, 1971 SIIFLIx)N C. SOMMERS. M.D.. Chairman Rrirarch Dirrctor, The Council for Tobacco Research-U S A, Inc. Director of Laharatorits, l.enox Hill Hospital Clinical PmJtswr of Pathology Columbia University College of Physicians and Surgeons New York. New York NOWARD B. ANDERVONT. Sc.D. Scientific F.ditor (retind), The Journal of the National Cancer Institute Bethesda, Maryland RICIIARD M. BINO, M.D. Director of Cardiology and lntrmnwal Mrdit inr Hunlington Memorial Flodpital, Paaadena, California Pro/rtsor of Mtdkine University of Southern California School of Medicine Los Angeles, California McKEEN CATTELL, PH.D., M.D. Professor Emcrltru oJ Pharmacology Cornell University Medical Colkge, New York. New York ROBERT J. IIUEBNER, M.D. Chic/, Viral Car+rinogenesis Branch National Cancer Institute Bethesda. Maryland I-EON O. JACOBSON. M.D. Dean of the Divirion of BioroRiral Sciences Rrgrnstein Professor of Biological Sciences University of Cfiiugo, Illinois CLARENCE COOK LITTLE, Sc.D.. LL.D.. LtrT.D. Scientific Director. The Council for Tobacco Research-U S.A., Inc. Director Enuritus, Roseoe B. Jackson Memorial Latxxatory Bar Ifarbor, Maine CLAYTON (;. LOOSLI. Pst.D., M.D. Hastints Professor of Medicine and Patholoty University of Southern California School of Medicine Los Angeles, California KENNETH MERRILL LYNCH, M D., Sc.D., LL.D. Chancellor and Professor En.rri(us of Pathology Medical Colkge of South Carolina, Charleston, South Carolina WILLIAM F. RIENNOFF, ltt., M.D. Profrssor Emeritus of Surgery lohns flopkins University Sdx)ol of Medicine. Baltimore. Maryland N~Y Q, ROBf?RT C. IIOCKFTT, Ptr.D. Acting Scientific Dbrctor m 1. MORRISON BRADY, M D. JOHN H. KREISIiF?R. Prt.D. 0 Asscxiatc Sci.nto/4c f)irerk+r Associatr Scientific )hrrrtor m VIN('FM F. LISANTI, D.M.D. tn rj ID Sc{rwN/Sc Associate I . (:LAREr1CE C(lOK L17TLE, Sc.D., I,I,.II., l.iu.l). 1888-1971 Dr. Clarence Cook Little, internationally known geneticist and cancer researcher, died December 22. 1971. lle had been Scientific I)irector of The Council for Tobacco Raearch-U.S.A., Inc. and its predecessor since shortly after the latter's establishment in 1954. 11e also served for many years as Chairman of the Scientific Ad- visory Board, an elective position for which only Board members vote. Former president of both the University of Maine (1922- 1925) and the University of Michigan (1926-1929), "Pete" or "Prexy," as he was known to colleagues and studenls, was a tower- ing figure as a man and as a scientist. Warm, dynamic, an en6agins personality with a sharp wit, Dr. Little was one of the fint scientists to discover evidence of a viral agent in eancer, an observation that led eventually to the generalized virua theory of cancer. He also did pioneering work in immunology and was one of the early re- searchers into the genetic eRects of radiation. Dr. Little founded the workf-famed Jackson Laboratory in Bar Harbor, Maine, in 1929. He served as its director until 1956 and as director emeritus until his death. lie 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 numerous other scientific societies. He uthortd many books and articles and his 1939 book, "Civilization Against Cancer," is still considered a classic for the layman. For 17 years Dr. Little gave The Council inspired scientific leadership and guidance. Ilis ccxHributions were many and important. He will be deeply missed.

Iritroduction CONTENTS •he Cancer Progam . . lCardiovaseutar Rcsearch I('hronic Respiratnry I)iseascs INeurophysiotogy. Psychc.physioloRy nd Pharmacology . Other Studic-t Abstracts of Reports . . . . . /Psychophysiological Studies JCarcinogencsis Studics . .C'ardiovascutar System . lRespiratory System . f'issue and OrRan Cul(ure . 6 . II 14 . 16 . IR 20 20 23 14 42 . . . . . . . . . . . 55 Studies at the Cellular Level . . . . . . . . . . 56 JPharmacoloRy and Psychopharmacology . . . . . . . 59 J Metabolic Studies . . . . . . • • • • • • • • 69 .rhcmistry and Biochemistry . . . . . . . . . . . 71 JEpidcmiolo6,y Active Projccts . . . Completed Projccts . 76 79 . 89 I Previous Annual Reports have stated the general research policics of The Council for Tobacco Researeh-U.S.A., Inc. as conceived by the Scientific Advisory Board. These policies, which continue to guide the program, emphasizc study of the etiology of those diseases that arc reputed to be related statistically in their incidence to cigarette smoking. Such diseases, especially eaneer, cardiovascular ailments and chronic respiratory afllictions, are not only leading causes of morbibity and death since the conquest of major infectious diseases but are universally recog- nized to be of multifactorial origin and to be strongly inNuenced by con- genital predispositions. The role of tobacco use in their etioko6y, if such exists, can therefore be defined or measured only within the context of a growing comprehension of the total etiologicd picture, invols•inR knowledge of the crxnple>< interactions between endoaenous and eaoRenous factors. As new concepts develop with respect to etiokgical mechanisms that may be involved in the genesis of these constitutional diseases, these con- cepts in turn suggest new biob6ical test systems tor determining whether chronic exposure to cigarette smoke or some ingredient thereof can con- trib ite 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 etiological understandin6 advances, to supersede the crude and inadequate methoda of the past. All investigators in the Ikld are weU aware that scientific progress takes place "in slow motion." The description of progress in a singk year is akin to attempting to depict the progress of a baseball game by describ- 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 atudks throughout the scientific world, the report of any one program can hardly hope to present anything resembling an integrated picture of the status of knowbd6e in any particular field or area of medical science. ~ Recognizing these limitatiom, we have attempted only to meotioo in this report a few sekcted Bndinas that have emerged in published form in the year ended June 1971. These selections do not in any way minimize the potential importance of many contributions by other investi6aton that do not receive particular mention here. Since all these contributions have entered the "public dcxnain" of acknoe, they will br assimilated into the body of total knowledge and comprehension that occurs through the process of "diRestion" and anabolism by the great confratcrnity of investi- 6ators in the many disciplines involved. Roaear t:' Ikx Kc r r rn ~ Acting Scientific INrector m 5 ~ m cv w

I .- ri'he Cancer Program Viruses and Cancer Viral aspects of nncer, inciudina the human disease, are being in- tensively studied in many labor.tories throughout the world. Rcports of advances in this active field seem to appear alrm»t weekly as scientists strive to pin down esact evidence that human nncrn, or at least some of them, are caused by viral factors. Perhaps of greatest prominence in this field is the general vinl 6enome concept. This postulates that c C-type RNA viral Rcnomc exists in .11 mammalian cells but remains "switched oft" until, for some reascn or reasons still largely unknown, it becomes activated or "derepressed" and triggers the development of cancer in forms determined by genetic programming of the strain or specics. I)eoelopnsent o/ a Test System A Gouncii-sponsorcd study has been aimed at development of super- sensitive, virus-primed tissue-culture test systems for measuring ahc relative potentials of various chemicals, in collaboration with the virus, to produce "malignant transformation" of celis. For exampie, rat embryo cells infected with Rauscher leukemia virw have been treated with various 3ows of 7/12-D+methylhen:(a)-anthracene (DMBA). Morphologic traisforma- tions indicative of noopfasia were seen. On the other hand, there was no transformation in uninfeeted rat embryo cells similarly treated or in infected cells not treated with tha DMBA. When transplanted into newborn rats, the transformed cells produced subcutaneoui sarrnmas, but no tumors resuited from like treatment with the infected or DMBA-treated, untrarrsformed cclls. The work showed that in this nodel, at least, both chemical and virus were necessary for cell transformation. Aceordina to the irwestiRator. the findinRs suggest that the C-type RNA viral genome of the Rauscher leu- kemia virus provickd specific oncogene information for the malignant trsnsformation. 6 I I It must he recoanixed that this model does not duplicate real life situations. Rather it exaggerates certain faetors and eliminates others for the sake of supersensitivity. The effects of chemicals on cell transformation are 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- pected to show positive effects by chemical agents which, on the basis of earlier 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 rrew tool that provides comparative ea- perimental values on a single suk. From many such evaluations a relative "activity scale" or act of scales may develop that will permit more direct and more nearly quantitative comparisons of very feebly active compounds nd mixtures with thosc 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 speeies, under defined, controlled and closely anal- agous conditions. A more distant goal is development of a method for assessing the relative degrees of cancer susceptibility in human patients. Another virus-related 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. The scientist who carried out the study reported that while C-type viruses in an active state have been found in the domestic cat, there is no evidence that they spread to doas or to man or even from cat to cat. It appean, under natural corrditions, that feline C-type viruses are inherited as indigenous latent Senes which may activate in the fetus or later in life. The investigator cautioned that susceptibility of certain human cells to in virro infection with these C-type viruses does not imply infection in the living organism. The same scientist, reporting on another facet of the study, disclosed indications that feline sarcoma virus can transform human osteosarcoma cells. This example of cell transformation by a non-host virus seems of considerable significance in virus-cancer research. Findings gained by this investigator in the (•nuncil-supp)rtcd project were applied to later research, funded by the National ('ancer Institute, in which human cancer cells were injected into the tetuees of preRnant cats Some of the cat fetuses developed tumors of human cells in which were 7

found particles similar to the known C-type cancer virus particles of several animal species. Present immunolo6ical evidence suggests that these may be the long sousht human cancer virus particles. If this finding is eonfirmed, it should greatly facilitate testing in man the many implications of the viral genome ccx+cept of cancer etiology that appear to present a rather wcU- integrated picture in several animals. It might also make possibk the addi- tiext of virus-primed human cell cultures to the in vitro test system that has been described. Other Ylrasal'..r+cer StrdlR. Lymphcxarcoma in r>,bbits, a rsre tumor In that particular animal, has been the subject of another Councif-sponsorid project. A rcxarcher studied one strain of nbbits amons..hich a number had developed lympho- saroomas. Ffe .nalyxed the pedigrees of the affected animals and found that inheritance indicated an autoaomal rectasive Sene conferring suscepti- bility to lymphosarooma. The flndin6s, the investigator rated, were con- ceptually compatible both with genetic susceptibility to malignant lymphoma and with vertical transmiaion of a virus. The neoplastic invofvement of various orRans in the rabbits resembled in many ways visceral lyrnphoaareomatosis of cats which has been proved to be caused by feline leukemia virus. According to the investi6ator, his findings on lymphosarcoma and its hereditary basis pmvidc a new and important model for studies of the pathoseneais of neoplasia. Still other studies being supported by The Council in the virvs area include: oncogeny and the antiviral action of interferon, eRects of respira- tory and oncogenk viruses in organ cultures of human respiratory tract tissue, and mechanisms o[ suppression of cellular immunity by carcinogenic hydrbcvbons. Smoke E:poaure Deolte. In its search for mexe realistic bioassay systems. The Council har for many years supported studies involving eiaarette smoke inhalation by animals. A major aspect of this undertakina is, of eourse, the drsi6n and production of a"smoke exposure device" that permits exposure of experimental animals to smoke inhalation in a manner and under amdi- tions that resemble human smokint sy eb.ely as possible. Previous Annual Reports of 7?w Councit have outlined severtl strict criteria for the function of such devices and these have subsequently been extended. The Council has developed a machine which has been undergoing tests in actual projects. While it 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 ditTicult problem is to measure how much of the particulate phase of the smoke actually reaches the lung surfaces of animals that normally breathe through their noses. Such measurements are being carried out with tagged smokes. The machine now being use-tested by several Council 6rantees has provision for insertion of a Cambridge filter that removes particulate material and thus permits comparison of the effects of whole smoke inhala- tion with those of inhaling the Ps-vapor phase akone. Handling and St.nd.rdisstlow of Anian.[. It is important to bear in mind that handling animals and confining them in a smoke exposure device ia very stressful to them. Hence "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 unhandled "caae controfs." Comparisons of these two sets of controls can be expected to disclose sonx of the biobycal effects of these particular stresses. These, at leaat, can then be taken into account in the interpretation of smoke inhalation experiments in which handling and eon6nement stresses cannot be separated directly from smoke effects. Sitnificant biologicd effects of stress, originally observed in such "machine oontrols," have now become a subject of investigation in their own right as part of The Council's program. In connection with smoke inhalation as well as other kinds of studies. The Council has sought improvement in the standardization of animals. Animals used in most Council-supported research are delivered by cesarean section and are substantialy free from patho6ens. Alw, their virus profiles are determined as fully as possible aa well as the status of the C-type RNA viral genome expression. Many cancer researchera uae inbred strains of mice because of the great variety of strains with known congenital tumor susceptibilities. Ho.vever, because thert sometimes are advantages in using hamsters, which have larVr funas and greater blood volume, studies have been started with this species. Some inbred hamsters strains re now available. 8 9

Expo.ure of (;ultr.res to Srwo4e In one smoke study, three types of primary cultures from Snell mice- kidney tissue, cmbryonic lung organ and lung explants-and one establ•shed cell line from Swiss mice were used to assay the biological eRects, in virro, of the gas phase from charcoal-filtcred frxsh cigarette smoke. Repcated exposure to puffs of the smoke did not produce any significant chang:s in the Snell mouse cultures. The exposed Swisa mouse cells showed art en- hancemcnt of 6rowth, mitotic index, and cellular atypism. No morpholcgical cell transformation was observed In any of the exposed cultures. Such in vitro studies are mainly useful as aids to the interpretation of whole animal studies. Being by necessity "unphysio{o6ical," they cannot be extrapolated easily or directly to rsormal life conditions. The researcher who performcd this study under a Council grant had previously reported that Snell mice, with viral genome expressed, exposed chrbnically to inhalation of either whole smoke or its gas-vapor phase (devoid of the particulates or "tan") developed lung adenomas and adenocarcinomas of the same kind that they develop spontaneously. None of the animals developed squamoua cell lung tumors. Mice of another strain, lackins viral genome espmsion, showed a dramatically lower incidence of adenoma and ade+xxarcinoma under the same conditions. In a study of thc incidence snd appearance of lung tumors and pneumonitis in mice, a Council-supported scientist immunized mice with an influenr.a virus and then exposed them and control mice to sub-lethal and highly lethal nebulized clouds of fresh virus. The controls succumbed quickly to the lethal cloud of virus and had extensive and persisting pultrronup changes following exposure to sub-kthal clouds. Tlse immun- ixcd mice becsme 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 exposed to airborne influenza virus. It is known that many mouse stnins develop adenomas in the bron- chiolo-alveolar regions regardless of what Is, or ia not, done to them. Adenomas do rsot seem to arise in the large bronchi or in the trachess of these animals. Is there a difference in susceptibility of these tissues to chemical agents? Or is it that these Inciting atcnts, when used cxperi- mentally, do not reach theae different tissues in equal amounts? In n effort to learn some rnwen, The Council has supported work in which fragments of tiarue from lungs, large bronchi nd tracheas, re- spectivcly, of HAI 8/c/Cb/Se mice were impregnated with methyl:ho- lanthrene (MC) alone or mixed with Wc powder in crxtcentrations of i:10 i and 1:100. The fragments were then transplanted into the subcutaneous fat pads of syngeneic hosts- Rcsults indicated that the sensitivity of bronchiolo-alveolar epithelia to MC was higher than that of tracheo-bronchial epithelia. However, the latter 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 ltscheal, laryngeal, bronchial and lung tissues, followed by a single intraperitoneal injection o( urethane, produced adenomas only in the grafted lung tissue, showing a tissue specific cflect of thia particular agent and suggesting lhat the agent was Iransported by systemic routes to the locus of action. Other Cancer Stud:e. In other work related to caneer, The Council is continuing support of eRorts to develop "positive" rnodels for the production of squamous cell lung cancer in animals. Guide lines are few in this arca, but if some simple, easily reproducible method could be devised, it would be very helpful in many research areas. If lung cancer of the type reported preva- lent in man could be produced repetitively, at perhaps the 20 percent level, in an easily handled animal, it would then be possible to undertake studies to see whether many fadora-age, sex, hormones, irritants, diet, atrcu, infeclion, and others - might influence the kvel and rate of incidence of the tumor. Also being continued is a project in which a concentrated stream o( fresh cigarette smoke is being blown directly on the skins of mice of a strain previously used in a skin-painting test with tobacco-smoke con- densate. Among other ongoing projects being supported are: hetero-trans- plantation studin with human lung ancer, the pathogenesis of urethane- induced lung adenomas in mice, and host factors in lung cancer and other lung ailments. Cardiovascular Research Ensynae ReIatkd to Atlrtrosr[ero.t. A study involvinR an enzyme has turned up findings that may hc significant in regard to atherosclerosis. 7he Council-suppcxted project used II 10

segments of hunian iliac arteries incubated with human serum £amples were heated to inactivate the enzyme lecithinrholoternl-acyl-trar sferase (1-CAT). It was found that when LCAT enzyme was present in the incubation rmdium, virtually only free cholesterol left the arteri:l wall. However, when LCAT enryme was absent, only cholesterol ester teft the wall. The rcsults suggcst that a decrrase in serum free cholesterol, caused by a serum cholesterol esterifying ertzyme, possibly LCAT, might be important in promoting the removal of free cholesterol from the arterial wall. According to the investigators who performed the work, the results are consistent with findings that there is a tendency for LCAT act.vity in the serum to be higher In persons with hypercholcsterolemia Rreatcr than 300 mR/ 100 ml, which may reflect a nxchsnism for prntccting'thc arterial wall from exccssivc free cholesterol. The scientists said their findings may be particularly important because they have found that cholesterol esterification is sip,niixantly impaired in persons with acute myocardial infarction and chronic coronary artery disease when compared with age-matched oontrols. Thus, they said, a deficiency of cholesterol nterifying enzyme, possibly I-CAT, may be important in the etiology or rate of devclopment of atherosclerosis. Coronary Mitrocircrlation 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 capillaries. Ne has described apillary blood flow as thal 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 eirculation. . To measure the velocity o( capillary red cetb, the investigator devel- oped a method showing the frame-to-frame pr)grr:s of individual red cells in moving pictures of the capillaries. The technique also permits measure- ment of the diameter of capillaries u it changes from frame to frame. Data from one part of the project iodicated that norcpioephrine re- suited in opening of capillaries, that nitrogfycerine slowed the flow in the capillaries, and that methacholine resulted in the complete arrest and even retrograde flow. While interpretation of theae findings will depend on con- tinuing study, the rtsardKr believes the work will provide new information bout the coronary microeulatbn. Another arpeet of the study wu a further elaboration of previously teported findings on the measurement of total and nutritional coronary I flow by means nf a coincidence counting system using rubidium-84. Workin8 with this system, the grantee found that norepineprine caused a dispropor- tionatcly greater increase in total as compared with effective flow, and the same result also occurred with nicotine. Isoproterenol, on the other hand, Icd to a proportionate increase in both nutritional and effective coronary flows. In another Council-supported study, mongrel dogs were cxposed to c~garette smoke inhalation and also to injection of various pharmacolosic agents. A reduction in aortic blood pressure during smoking by the anes- thctized animals was seen. This was attributed in part to the effects of nicotine on the central nervous system and in part to vauxfilation caused by the acticxt of rcleased catecholamines on certain receptors in the pcri- phcral vascular system. Kinina and /nJ[amnuttory Reactiona Another investigator has been studying kinins, polypeplides 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 propertics of the components o( the kinin-forming system in rabbits re 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. Framing/aam Heart Study The Council has made a ttant related to the Framingham fleart Study, effective July 1, 1971. This study was to have been ended for lack of federal funding, but its long-time director and several associates sought and obtained outside support to keep it going. 'iTre Council's grant is for an investigation of smoking elasses, risk factors and cardiovascular disease in the population being followed by the project. Other studies being supported include: nicotine and various aspects of cardiovascular function, carboxyhemogbbin and cardio-respirstory funetion, smoking and air pollution in human myocardial metafxolism, in- fluence of smoking and nicotine on lower limb cilculation, sensitivity of vascular tissue to nieotine, and the effect of smoking on rcRi(wul ccrcbral blood flow in smokers and nonsmokers. 12 13

Chronic Respiratory Diseases flyaline Membrane Dfuase A sianificant finding in regard to hyaline membrane disease (ItAf)) has conx from a Council-supported project that encompassed the anrlysis of 387 autopsies on newborn and stillborn infants. It was found that the adrenal glands were 19 percent liahter in infants with HMD than in those free of the disease. TTrose without the disease had a greater numb-r of adrenal cortical cells. Also, a positive correlation was found between the presence of infection arising before birth and the absence of IIMD. In connection with surfactant, which appears to play an important role in this disease, it is Interesting that anencephalic neonates who had little or no adrenal fetal cortical zorxs and half-siz.ed adult zones, were found to have only 45 percent the mass of osmiophilic granules in pul- monary type It alveolar cells found In non-anencephalic control inlants. These granuks reportedly are the anatomic representation of sur/a<tant. The investigators who eonductcd the study noted that the corticcnt,roid mechanism "oflers the ttractive possibility of relatively simple prophylaxis against hyalinc membrane disease in certain high risk gestations." But, they caution, "This temptatiixt to treat should be strongly resisted until the mechanisms in qucstion have been explored in the good experimental models of hyaline membrane disease now available. The need for such, caution is also supported by the observation that the doscs of cortico- steroids and ACTFf used to increase surtactant levels in the lungs rrf fetal lambs have induced toxic changes in other organs." AppaiIatAian Coa! Miners A quantitative morphdoye atudy was undertaten of lung and heart structure in 322 Appalachian minen who were classified by age, duration of mining exposure, rank of coal mined and smokin6 habits. Right ventric- ular hypertrophy as evidence of cor pufmonak was common after age 50. The volume of macular dust lesions, includin6 asxsciated silica crystals, fibrosis and focal emphysema, increased with age but these dust lesions were not deemed to be solely responsible for dyspnta since miners with dyspnea also had a gcneralired form of emphysema. Smoking was associated with a modest increase in bronchitis and emphysema in bituminous miners but showed littk or no relat;on to the extent o( such lung conditions in anthra- cite workers, and ninunioltins miners also developed these disorders. Ah)ve-ground air pollution may play a role since the wives of sonx bi- tuminous miners seem to be nearly as much*aReeted as their husbands by cough, phlegm, wheezing and breathlessness. The association of prxu- moconiosis with coal rank was found to be fortuitous. Microcirceelation o/ the Hurnan Lung During the year a Council grantee published a report describin6 the blood and lymphatic microcirculation of the human lung that provides a clear, concise summation of major findings in this particular field. The investigrlor, long involved in lung research, gave the results of correlated macro.arpic and microscopic studies o( the lungs o( humans, beagle dogs and mongrel dog.. The relxri was primarily concerned with angio-arehi- tectural relationships between the bronchial and pulmonary vascular sys- tems, and the author noteQ that the relationahips are distinct in each. The bronchial arteries are dirtxtly connected to each other, establishing a "parallel" vascular supply. 'I"he bronchial veins also form the peribronchial ven ws plexus. However, the pulmonary cireulation, the pulmonary arteries and veins form terminal, Intralobular branches. The bronchial and pulmonary arteries also communicate in the bronchioles through a common capillary network. The grantee reported he has been unable to observe broncho-pul- monary arterioarterial or arteriovenous 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. Ile said they probably vanish in child- hood but reappear in the adult lung with the onset of even minimal pul- monary inflammatory reactions or processes. The scientist also published another report in which he described cells with the ultrastructural characteristics of neurosecretory cells in the lung lobules of prematurely born infants. He had previously suggested the existence of these cells on the basis of light and fluorescence microscopy and noted that analogous celis had been reported by othen. Ife said the possibility exists that these cells may be involved in the regulation of lobular growth and in modulation of normal perinatal cardiopulmonary adaptation, and that they may play a role in respiratory distress syndrome. Pulnsonory Al orofor M.croplsa6es What is the origin of the pulmonary alveolar macrophaRrs? "fhcre has been considcrable controversy as to the cellular precursors of these cclls, which comprise a major defense mechanism of the lung. A Council-sup- 14 15

ported study has elicited evidence to indicate that at least some of the macrophsges originate from bone marrow. The study used genetically related mice and substrains to investigate the possibility that macrophages originate in marrow stem cells. Success of the experiment rested on the fact that the genetically re- lated mice carry a nonspecific, readily identifiable pre-albumin esterase marker. Lethal x-irradiation and injections of bone marrow from the mice led to findings confirming that at kast some macrophages originate in bone marrow. This particular work waa part of a project that is moving into the biochemistry of macrophagea. Another Council gantte provided a deacriptive report of the appear- ance under the ekctroo mkroaeope of alveolar macrvphagiea obtained by endobronchial lavage from smokers and eoeumoken. He said that in yeneral matrvphages from smokers contained larger and more heterogeneanly strucfured Inclusions than those lrom noo- smoken. However, there wu considerable variation in the number, size, and heterogenicity of indusions within individuaf macrophiges from smoken, suggesting that various stagra of phagocytosis were represented in lavage samples. Further, the researcher said, sorne substano-s that are phagocytized but not digcsted probably aecvmulated in the larger hetero- geneous inclusions of smokers. Other studies o/ lung macrophages have shown that a greater number are recovered by lavage from smokers than from nonsmoken, nd that the macrophages from smokers show unimpaired phagocytizing capacity in vitro. , Neurophysiology, Psychophysiology, and Pharmacology Nicotine and Memory Conao/1d.tlors For several years The C,oaroeil has been supporting research into the effects and .ction of niootine on eremorY eonsolidation. One researcher has found that rats put through a training regime (mase sdving and kvcr pressing) learn at a meuurr+ble rate. Fk+wever, if the animals are given an electric shock' after a training session, they tend to "unlearn" everything they had previously acquired. Apparently the shock caused an amnesiac condition and prevented consolidation of the karning process. When the animals were treated with nicotine prior to a training session, it was fuund that the subsequent electric shock did not affect memory conu.lidation; the animals largely retained what they had learned, apparently because the nicotine blocked any effect 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 suggests that the laltcr substance may be the active agent. Other parts of the project included testing the role of the serotonin precursor. S-hydroxytrypk>phan; age diffcr- cnces of the animals; Jifferential housing; reduced brain amine levels; and amine changes at the cellular level. In work in a related area, a Council-supported researcher found that chronic treatment with nicotine induced an increa-ud turnover rate and utilization of noradrenaline in the brains of rats. Pretreatment with nicotine did not affect the action of sedative, antidepressant and central stimulant drugs in the animali brains. In short, the chronic administration of nicotine appeared to stimulate 7he brains of the rats, making the animals function more effectively. Sex Ilorrnone.a and Emplryaeina A potentially significant report during the year came from a Council grantee who has been working on the experimental induction of emphysema in rats. 1fe has sought to determine whether such induced disease would kad to pulmonary hypertension and cor pulmonale as in man. It was found that daily injections of progesterone (the female sex hormone) and medroxyprogesterone (in a dose 1/50th that of progcsterone ) prevented experimental induction of emphysema in the rats by a method that other- wise was successful. Another grantee, treating rats daily with nicotine injections, found an elevation in urinary excretion of catecholamines that peaked after three days. After seven days of continuous treatment, there was a marked fall-oft in the urinary Ievels, and by 14 days the ealecholamine excretion was essentially normal. TAis suggrsts a physiological accommodation or adjust- ment over a pericxl or time to chronic administration c>< nicotine. The scientist explored three poasibk mechanisms that might be reslxxr- sible for the return to normal of the ekvated urinary cateclVamines. He concluded that tolerance to the nicotine-indueed elevations of urinary 17 16

catecholamines resulted from increased metabolic enzyme activity that in turn led to faster metabolism of the catecholamines released from the adrenal medulla and adrenergic nerve terminals. Other Studies r.on6-Tern. Strdy of veter.ns The Council haa been participating In a kmg-term study that en- compasses periodic cxaminatiorn o( more than 2,0(X) male veterans. Pur- pcne of the projeet, which is supported mainly by the Veterans Administra- ticm, is to investigate a broad range of changes that occur in initially healthy persons as they age. One ultimate purpose of the investigation is to karn what factors may be predictive of longevity or conversely, of early decease. In this study the investigatcxs became aware of "secularity effects" as widely encountered, complicating factors in studies of human populations that extend over relatively Mng periods of time. While in well-designed anlmal experiments environments can be kept quite constant except for the independent variable to be studied, changing cultural and technological aspects of human Gfe introduce time-dependent alterations that are often not taken into account. In a book published in 1971. the two investigiton conducting the project noted that when they learned substantial numbers o( centenarians were heavy smoken, they gave serious thought to the possibility that smok- ing was secular, rumely, "that the earlier death of those that smoked more and those that smoked cigarettes was due to the fact that they were born tater and lived at a time when there was more smoking, rather than because smoking was hfe-ahortenin8." Their findings, they said, sugaest that the correlation seen between smoking and age-at-death may at kast partially be explained by the secular increase in smoking. They further twted that in a study such as thcin, "in which all the individrab die essentially at a single point in time, one can- not precisely rressure the effect of smoking on longevity since the prob- ability of a given individual being a cigarette smoker or nonsmoker Is re- lated to the birth year." Secularity eReets, the authors suggest, rsay influ- cnce the results of many longitudinal human population studits. Riocfremicof E/Jects of Stre» In a continuing study supported by The Council of certain biochemical effects of chronic smoke inhalation by'guinea pigs, a researcher was able to detect effects of stress and smoke inhalation on lung mitochundrial phosphorylation. l hc nostrils of the animals were occludcd with warm, molten paraffin or a quick-setting, liquid silicone rubber to insure mouth breathing. Sham-exposed animals were subjected to nose plugging, con- finement and machine noise without smoke. The stresses of forced mouth breathing, handling, confinement and noise resulted in losses in oxidative phosphorylation efficiency in the guinea pig mitochondria. ffowever, such loss of efficiency was considerably re- duced by cigarette smoke inhalation. Nicotine is not the active agent in this effcct. Twin Studies With financial aid from The Council, an international symposium on twin studies was held in San luan, Puerto Rico, December 1-4, 1969. A report on the symposium, titled "Twin registries in the study of chronic disease with particular reference to cardiovascular and pulmonary disease," was published in Acta Medics Scandinavica, Supplementum 523. 1971. The report discussed research into smoking that involved twin regis- tries in Sweden and the United States. It said the clinical twin studies "do indicate a presence of genetic factor in coronary heart disease and in variables which have been found related to it. For instance, blood prrssure, serum cholesterol, and triglycerides tend to be correlated between the members of pain, and in addition post-exercise ST-dcpressions of the ECG were the same in the twin pairs regardless of smoking." The report further said: "A statistically significantly higher mortality among smokers was shown in dizygotic male pairs, born in 1901-1925, but not in monozygotic male pain, or in femaks of either zygosity. No In- creased mortality was apparent among smokers in the older age-groups (1889-1900)." . Also in 1971, a sKcond suppkment to the 1961 monograph, "Tobacco - Experinxntal nd Clinieal Studies," was published. This was an update of the colkction, abstracting„ and analysis of the world literature on tobacco and includes published reports on both experimental and clinical studies. T.,e Council has supported this project throughout. 19 18

Abstracts of Reports Following are ahstracts, apDroved by the authors, of reports on new expcrimental research aeknowledging support from Thc Council that have appeared in scientific journals since publication of the 1969-711 Report. 7 he name of the recipient is in italics. The abstracts are grouped under these headings: 1. Psychophysiologi- cal Studies, I1. C'arcinogenesis Studiea, 111. Cardiovascular Systern, IV. Rcspiratory System, V. Tissue and Organ Culture, V1. Studies at the Cellular Level, VII. Pharmacology and Psychopharmacology, VIFf. Me- tabolic Studics, IX. Chemistry and Bioehemntry, and X. Epidemiology. 1. Psychophy.toloRtcol Studies PSYCHOLOGICAL ASPECTS OF CIGARETTE SMOKING IN MF:N: A CLINICAL EVALUATION Certain psychological factors influence smoking behavior patterns; psychological factors, the sanx or others, are also involved in the discon- tmuance of the habit. In this study, heavy cigarette smoking in men is theonlud to be associated with an oral-impulsive penonalily constellation. Clinical interviews with 130 subjects, representing non-, former, mild, and heavy amokers, supported previcws questionnaire findings that thc last named group is characterized by features of: defiance, rmpulsiverxss, danger-seeking, neurotic labaity, and oral preoccupation. In (his sampIc, heavy smokcrs' relationships with their fathen were more disturbed than were the other groups' paternal relalionships. Intensive follow-up interviews with a small selection of subjects indicated factors associated with starting and stopping smoking and an alternate personality pattern in heavy smokers previously seen as false negatives on the questionnaires. This subsample handled underlyin6 oral conflict with traits of obsessiveness, guardedness, aqd cautiousness. Former smoken seem able to discontinue the habi: when pcrsonally nxaningful, prn itive events occur and/or when their health is actually threatened. Bolh life events and factors of personal resoutcefulness are relevant in understanding why some men can give up the habit and others cannot. Jacobs, M. A., Knapp. P. l/., Rosenthal. S, and Fiaskcll, 1). P.tychotonfaric Medicinc 32 ( 5):469-485, 1970. From the F-Svision of Psychiatry. Boston University School of Medicine, Boston. SOME VARIABLEiS CONTRIBUTING 1-0 TIiF ONSE?T Ul' ('IGA- RCTTE SMOKING AMONG )UNIOR HIGH S(-FI(X)1. STttl)FN1S By analyring tlutknt resporues to  detailed questionnaire. thir dudy systematically expforcd a subject's perception of the conditions and forces present when hc smoked for the first time. The sample consisted of a total of 2,729 junior high scho students., According to their answcrs, rc- spondcnts werc ranked as nonsmokenol ertPerimental smokers, or regular smokcrs. Results showed that smoking by junior high school students was partly a function of smoking incidenee by their prevailing models since one of the bcst predictors for children smoking is slill whether or not their parents snoke. Regular smoking adolescents appcared to have thc highest proportion of parents and friends who smoked regularly. The sociaF nature of smoking became quite obvious when the data were analyzed. Boys were generally either encouraged to smoke or actually smoked with other boys whik girls maintained the same relationship with other girls. Neilhcr parents nor siblings appeared to assume any direct role in the instigation of smoking. The age at which the first smoking experience occurred varicd somewhat for boys and girls. By age 12 most boys who were going to try to smoke had already done so while most 4ir1s had al least tried to smoke by the age of 14. Girls re/rained from smoking because of the possibility of parental disapproval while boys were afraid of its potential effect on their athletic skills. Changea in students' smoking status occurred in spite of their Enowkdge bout potential harmful effects and their assessment of parental disapproval. Thus the meaning ascribed to smoking bX the culture and its place in the adolescent subculture was of maximum rmportancc to the sludents. Pafiner, A. B. Social Scirnce and Medicine 4:359-366, 1970. From the Department of Psychology, University of Toledo, Tokdo, O. VISUAL DEPRIVATION AND PAROTID RESPONSE TO CIGARETiE SMOKING Smoking while blindfolded slgniBcaAlly and consistently reduces sa- livary flow rates compared to flow rates associated with normal smoking. In this experiment, cigarette smoking signifkantly increased parotid salivary secrelion in eight smokers and eight nonsmokers. Obstructing vision by wearing opaque gogglca consistently reduced the salivary response to smok- ing in all 16 subjects. Unstimulalcd resting levels were lowcr when vision was blocked, suggesting a more general phenomenon than merely removal of the sight of the stimulus-objecl. Smoking cigarettes through a Cambridge filter assembly, which trapped the particulate matter and rendered the exhaled smoke invisible, had no effect on salivary secretion. This indicates that the salivary stimulation derives from irritating materials in Ihe gas phase of the smoke. PanirAorn, R. M. and Sharon, 1. M. Phytio7ojry and Behavior 6(5):559-561, 1971. From the Fkpartnunt of Food Science and Technology, llniversity of California. I)avis, and the University of the Pacific Schtwd i f Ikntiary, San Francisco. 20 21

SOME CNARA(TFRISTICS RELATED TO TIEE CAREER STATUS OF WOMEN PHYSICIANS Between the years 194R and 195R, 81 women graduated fmni ihe Johns Hopkins Medical School, often with grade records surpassing those of their male classmates. Of these 81 women, 69 were in active prac ice in 1967. Over one-quarter had ttained specialty certification and 219 held professional appointments; these two criteria were taken as a crude m.:asure of career success. Two factors related to career status were marital status and family size. BeinR married, especially at an early age, was associated with a less successful career, as was increasin8 family size. Althourh nearly 78% of these graduates had married, the most successful women, the "pro- fessors" 8roup, were most likely to be single. They were also more likely to be regular cigarette smokers and to have lower scores on "habits ol nervous tension" than their colleagues. In these two respects the) were similar to their mak medical school co(lea(Ives. Daughters of managers and proprietors had the best arades and most Qrominent careers; daugh.ers of physicians were rnost likely to be In practrce. It is suggested that f,art of the reason women medical graduates do not realize their full potential in later careers lics in the views and practices of society. Westlin8-Wikstrand. EI., Monk, M. A. and Tho.nas, C. B. The Johns IropAint Mrdk-al Journal 127( 5):273-286, 1970. From the Department of Environmental Medicine and Community Efealth, State University of New Yor1t, New York CSty, and the Department of Medicine, The Johns E(opkins University School of Medicine. Baltimore. PERSONALITY CHARACTERISTICS OF MEDICAL STUDENTS AS REFLECTED BY T11E STRONG VOCATIONAL INTEREST TEST WITEI SPECIAL REFERENCE TO SMOKING EIABITS The Strong Vocational Interest Blank (SVIB), a psychological mcas- urrment toof, was administered to Johns Nopkirsa medical students in seven consecutive classes graduating in the period 1958-t964. A total of 383 white male students with known snwkin8 habits at a mean age of 23.2 years completed the SVIB. Using the SVIB and smoking mcasurements, discrim- inant function analysis for two criterion poups was carried out by means of a multivariate program. Results showed that smokers and nonsmokers had significantly different personality characteristics as measured by the SVIB; this overall diRerence was highly significant. When single SVIB variabks were compared, light ciprctte smokers shared some of the main occupational interests of the heavy smokers. but had distinctive differences as well. Pipe smoken were significantly different from nonsmokers in regard to another set of single ocupational variables. In 1968 the smoking habits of the test group were rechecked and it was found that less than a third of the original group of heavy cigarette smokers had stopped smoking. 'Elsc group of physicians who had stopped smoking more closely resembled non- smokers in their vocational interests while in medical school than did those who were continuing to smoke. Thomas, C. B., FarBo, R. and Enslein, K. The Johni flop4ins Medical Journof 127(6):323-335, 1970. From the Department of IHedicine. The Johns Hopkins University School of Medicine. Baltimore, and The Genesoe Computing Center, Inc., Rochester, N. Y. CFfARACTERISTICS OF MALE MEDICAL STUDENTS RELATED TO THEIR SUBSEQUENT CAREERS This study of 724 1948-1958 male paduatca from Johns Ifopkin. Medical School shows that academic standing and age at 8raduation were closely asuxiated with professional careers. The t~re of practice, the attainment of specialty boards, and to aome extent the academic position of those on medical school faculties were all related, sometimes in a com- plex way, to class standing and a8e. Other factors such as tathcr's oceup.- tion, ethnic background, type of college attended, and son,c selected psychob8jcal and physical measures were not signiBcantly related to later careers. Marital status during medical school seemed to have sonre rela- tirxrship to later achievement reprdless of academie standing and age. In this study medical school grades were definitely related to later achievement for graduates. It is not clear whether or how grades reflect later ability but they do seem to influence careers. That grades do influence opportunities for graduate training and careers suggests that thest grades should be based on courses and training adequate to the variety of activities which physi- cians are now expected to perform. Monk, M. A. and Thomar, C. B. The Johns Hopkin.r Medical Jounwl 127 ( S):254-272, 1970. From the Department of Environmental Medicine and Community Health, State University of New York, New York City, and the Department of Medicine. The Johns Hopkins University School of Medicinc. Baltimore. 11. CarcinoRene.la Studies T()MORI POLMONARI DA IDRAZINA SOLFATO A DOS1 RIDOTTE IN TOPI BALB/c/Cb/Se OONADECTOMIZZATI Administration of hydrazine sulfate Increased the percentage of pulmonary tumon in Conadectomized, intact virpn, and breeding female mice. Tumors were Induced by hydrazine sulfate in the same rrequency in 8onadectomiz.ed mice and in intact virgins. Endo~e n ovarian stimulation associated with hydrazine sulfate induced in breedcrt pulmonary tumors more malignant than those seen in intact virgins. Without hydrazine sul. 23 22

fate, the incidence of pulmonary tumors in gonadectomhed 13Ai_fi/c/ C'b/Se mice was as low as it was in the controls. Bioncifiori, C. Lav. Anat. Pat. PeruRia 30(2):113-119, 1970. Other support: Anna Fulkr Fund. From the Division of Cancer Research. University of Perugia, Italy. T(1MOR1 POLMONARI ED EPATICI DA IDRAZINA SOLFATO A DOS1 RIIX)TTF. IN TOPI BALB/c/Cb/Se In these investigations, the incidence of spontaneous pulrionary tumors was low (24% in males, 4% in femaks) whilc that of liver ~unrors was urn. Ifydrarinc sulfate administered in reduced drnes increased the frequency of pulmonary tumors as follows: 0 56 mg (total mR R t), 65 % in males and 76% in fcmaks; 0.2e mg (total mg 42). 62% and 89%; 0.14 mg (total mg 21) 54% and 32%; 1.13 mg (total mg 32). RS 76 and 75'f6, and induced liver tumors: 0.56 m6, 7% in males and g% in fr males; 0.29 mg. R% in maks. When the average induction time of pulrnonary tumors was longcr, liver tumom were also observed. MorpholoFicr Ily the pulmonary tunxxs were adenornas and/or carcint.mas; thc liver :umors were highly vasculariud hePatocarcinomas. The possibility of ir ducing F ulmona7 nd liver tumors in mice with per-kilo dosts of hydrarine sul- ate, equivalent to thosc pcr-kilo derived from the adminisaration of isoniarid in man, is important for its applicability to the use af isomiarid in human pathology Biancifiori, C. L.ov, Anat. Pat. PeruRia 30( 2): E9-99, 1970. Other arpport: Anna Fuller Fund. From the Division of Cancer Research, University of Perugia. Italy. OVARIAN INFLIIENCE ON PULMONARY CARCINOGENESIS BY NYDRAZINE SULFATE IN BALB/e/Cb/Se MICE Alteration of the hormonal state of mice influenced the incidence of pulmonary tumors induced b7 hydravne sulfate (IfS). Daily administra- tion of IfS to BALB/c/Cb/Se female mice in various hormonal states varied the incidence and the average number of pulmonary tumors per tumor-bearing mouse as follows: in intact vir6ins, 90.0% and 3; in breedcrs, 1(x)% and 14; in gonadectomizrd mice. 60.0% and 5. Ftistologically, in intact virgins. 3.3% were carcinomas and 96.6% were adenomas; in brceden, 47.2% were carcinomas and 52.7% were adcrximas; in Rona- dectomired mice, 4% were carcinomas and 96% were adenomas. Many pulmcx+ary carcinomas induced in breeders infiltrated the thoracic wall and the mcdiastinal organa and rnetastasized to the adrenal glands nd myo- cardium. '1 he irx•reaud biologic and rntxpholoRic malignancy of pulmonary tumors seems to demonstrate that a greater ovarian hormone production. as occurred in breeders, accentuated the already existent susceptibility to pulmonary tumor induction in BALB/c mice. Whether this is asscxiatcd with an inhibition of the immunologic factors remains to be dctcrmined. 8iancifiori. C. Journal o/ the National Cancer Inttitute 45( 5):965-970, 1970. Other support: Anna Fuller Fund. From the Division of Cancer Research, University of Perugia, Italy. CANCER(x1ENFS1 DA IDRAZINA SOLFATO IN TRAPIAN"1I ISOrENI('I TRACFIEO-BRONCOPOLMONARI IN TOPI BALB/c/Cb/Sc Carcinogenic activity of hyorazine sulfate on the trachca, bronchus, and lung fragments of ncaborn BALB/c/(b/Se mice, transplantcd to the same strain, was studied ia this e><perinxnt. The lung fragments by them- selves showed 35% adencmas and 13% eareinomas. When the lung frag- ments were transplanted, )ydratine sulfate induced tumors, which showed greater malignancy, in a s7wrter time in the newborn lung fragments than it did in the lungs of the adult hosts. It would seem that this was caused by the marked susceptrbiiity of newborn mouse tissue to the carcinogen. Explanation of the findir.gs with regard to the divergent frequencies at which the trachea, bronc:ii, and lung parenchyma developed tunuors re- quires further research. Biancifiori, C. Lav. Anar. Par. Prrugia 30(3):137-146, 1970. From the Division of Cancer Research, University of Perugia, Italy. TRANSFORMATION AND PRODUCTIVE INFECTION OF IIUMAN OSTEOSARCOMA CELLS BY A FELINE SARCOMA VIRUS The studies reported here indicate that feline sarcoma virus ( FSV ) can transform human osteosarcoma cells; two lines (MT-1'SV and MT- FSV, subline 2) of transformed cells now have been established. When compared with the parent MT cell line, the FSV-transformed lines have, in early passage, an altered morphob~y increased sataration density, in- creased growth rate in 2% serum, and increased plating effxicncy in agar. The transformed cell lines contain C-type virus particks and feline C-type virus antigens not detected in the M'T line. Particles released from thc transformed lincs have a hoat range (do; and human cella) similar to that of the parent FSV. •Cclls of aubline 2 in seventh passage induced fibro- sarcomas in prenatally inoeulated kittens; the tumor cells, adjacent muscle cells, and submaxillary gland cells shed C-type particles. In a second set of experiments, the cells of the osteosarcorna cell Pne in passage 7 have been transformed by each of two stocks of FSV (the stock used in the original study and one derived from a line of FSV-transformed beagle cells). llrcx transformed cclls, as those in the first experiment, contained fciine ('-typc 25 24

virus antigens, shed particles with focus-forming activity on hcaRle and human cclls, and exhibited C-type vinis morphology and a density of ap- proximately 1.16 g/cc. McAllister. R. M, Filbert, J. E., Nicolson, M. O, Rongey. R. W. Gordnrr, M. R., Gilden. R. V. and Fluebner. R. J. Norurr New Biology 230(17 ):279-2g2, 1971. (hh.r.rpport: National Cancer Institute. From the Departments of Pediattica and Pathology. University of California School of Medicine, Lo. Angeles. REDUCED LYMPHATIC DRAFNAGE FROM HAMSTER CFI[iFiK Pfll>CH: A PROPOSED EXPLANATION OF ITS IMMUNOF-(X;ICAI.LY PRIVILEGED CFIARACtER To answer the question of whether or rsot a direct route exists Ixtween the hamster's check pouch and its regional lymph nodcs, the kft check M hes of eight golden hamsters were everted and injected with 0.1 ml of in. Serial sacrifices were performed at intervals of 2, 4, 6, and 24 hours post injection and the kft cheek pouches, kft and right su xnental lymph nodes, and livers were removed and histochemically stained. .\nother group of eight hamsten, serving as controls, received 0.1 ml of the ferritin solution intracutancously to the left part of their upper lips. These .nimals were sacrificed and their tissues processed as in the cheek pouch group. Histochemically demonstrable iron was prexnt only in a.few toc of the ipsilateral nodes in the group sacrificed at 24 hours post injection of ferritin in the cheek pouch. In eomParison, the same nodes already contained a greater amount and distribution of stainable iron at two hours after injec- tion to the lip. These results Indicate that while there is a direct route from cheek to lymph node, drainage via this route is reduced and delayed. Im- munolosically, this situation of  reduced and delayed drainage from the hamster eheck pouch affords foreign grafts in this site a decisive advantage for prolonged survival. Gofdrnbert. D. M. Exlxritnrla 26( 8 ) :907-90R. 1970. From the Department of Pathoiogy. University of Pittsburgh Sc',wool of Medicine, Pittsburgh. IIISTOGENETIC RELATIONSNIP BETWEEN CARCINOIDS AND MUCIN-SECRETING CARCINOMAS OF COLON AS REVEALED BY FIETEROTRANSPLANTATION A human colonic eeoplaun with morphologic characteristics of a carcinoid was successfully transplanted to cheek p~iuches of adult golden hamsters after a short ao}ewrn in cell-impermeable chambers in rats Al- though no mucin-secreting cella were detected in (he dimor carcinoid, the check pouch trsmplants exclusively exhibited mucin-seereting tumor cells of signet-ring type consistent with adenocarcinoma. This transplantable turnor, designated GW-77, has retained this appearance as well as expan- sive growth characteristics in xenogeneic hoats for a period of four years. The findings presented here represent strong biological evidence consonant with views advocating a hislogenetic relationship between colonic carcinoid and adenocarcinoma. GofdenberR, f). M, and Fisher, E. R. British /ournal oJ Cancn 24( 3):610-614,'1970. Other.rrportr U. S. Public Health Service. From the Department of Pathology. University of Pittsburgh School uf Medicine, and the Veterans Administration Hospital, Pittsburgh. RE[)UCE[) I.YMPHATIC DRAINAGE FROM HAMSi'ER ('FiEEK POUCH A transparent is+olation chamber permitting continuous observation of the hamster cheek pouch wu used to study the movement of injected India ink from the cheek pouch wall to the regional cervical ipailateral or contra- lateral lymph nodes. Th,e isolation chamber was constructed specifically to obviate the experimenta.^ error of dye leaking out of the cheek pouch proper to adjacent tissues. A brownish-black coloration of only the ipsilateral sub- mental lymph was first observed 20 hours after Injection; the coloration was definitely localized ¢ven though distinct lymphatic channels in the pouch could not be sa:n. Thest finding>t knd support to the thesis that there are lymphatics In the hamster cheek pouch. Compared to the time rsecet<sary for egression of dye from other areas of the hamster head, it appears that there is a delayed lymphatic drainage from the cheek pouch, by a factor of ten. In light of these observations, It is suggested that the so-called "immunolosically privileged" character of the bamster cheek pouch is related to a delayed and reduced lymphatic drainage from this site. f;oldenbtra, D. M., and Steinborn, W. Proceedings oJ the Society /or EsprrLnenta( Biology and Medicine 13 S( 3): 724-726, 1970. From the Department of Pathofogy, University of Pittsburgh School of Medicine, Pittsburgh. RETENTION OF HUMAN PROPERTIES BY A XENOGRAFTED IIUMAN COLONIC TUMOR, GW-77, PROPAGATED IN UNCONDITIONED HAMSTERS GW-77, a newly established transplantable neoplasm of human colonic origin, is capable of expansive growth In uneonditicxted, adult golden hamsters. l?tis unusual 6trowth-behavror of GW-77 raises a question as to the lurmx's spocics-idenldy. In the present study, this question was studied by immunofluorescent and biochemical means. Both the direct fluorescent antibody test and its lactate dehydrogenase isexnzymonwbJity 27 26

pattern rcvealed the human specics-specific character of GW-77. Thc rc- tcn/ion of its original sprcics•identity is consistent with GW-77's rclalivcly Icss malignant, only locally expansive, growth-behavior in various sitcs of the hamster. Goldrnhrrg, I). Af., Bhan, R. D. and Pavia, R. A. I'rocrrdinRs of the Scx rrly for Experimental Biology and Medicinr 135 ( 3): 657-659,1970. Other support: National Cancer institute. From the Department of Pathokgy. University of Pittsburgh School of Medicine, Pittsburgh. STATHMOKINF.TI(' EFFECT OF COLCEMID ON A PRF_SI1MPfIVF. IIl1MAN IIAMSTER IIYBRID TIJMOR, GW-47R -The action of Cokemid on the aacitcs variant of the (7W-47R tumor, originally a human stem-cell lymphoma propagated in hamster chcek pouches, was compared to its action on Fortner's hamster amclanotic melanoma A. Mel No 3. The two tumor types exhibited strikingly drflerent slathmokinetic responses to the alkaloid Colccmid. T-he percentage of un- trcaled sscitcs tumor cells in metaphase wa.s found to be 0.98±0 I' and 0.95!0.23 for GW-47RA and A. Mel. No. 3, respectively. The a-.oRee of inetaphasc arrest in the A, Mel. No. 3scites tumor cells occurrcJ at a dose of 60 mg/kg Colcemid, whik the pogee for GW-478A was at 20 mg/kg, These results, which were quite uncxpected, indicate that (;NI-478 ascites tumor cells resfu,nd to Colccmki more as human than as hamster cells. It appean, thercfore, that althcwgh GW-478, a highly malignant trans- plantable tumor in unconditioned hamstcn, predominantly has hamster-like properties, it lacks the S7rian hamster's genetically controlled resistance to this coichicine derivative. This further supports the hybrid character of this neoplasm and the usc of this trait as a genetic marker in hybridiza- tion experiments with Syrian hamster and other xenogencic cells. Coldrn6rra, D. Af. Experimental and Molrcular parhology 14(1 ):134-137, 1971. Other support: National Cancer Institute. From the Depa rtment nf Pathology. Temple University Health Scicnccs Center, Philadelphia. INNIBITTON OF MURINE SUBCUTANEOUS AND INTRAVENOUS BENZO(RST)PENTAPHENE CARCIN(X;ENFSIS BY SWEET ORANGE OI1,S AND d-LIMONENE Orange oils and d-limonene slow the rate of tumor formation following subcutaneous injection of benzo(nt)penlaphene (DBP) in C57B1./(i lax mice. 'fhes,e anticardnrrgenic rtia of the terpenes were discuvercd during the course r+f studies siEned to evaluate the pcnsiblc cocarcino- genicity of natural lerpcncs (citrus oils) and of pure d-limoncnc in sub- cutaneous injccticrn sites and in the lungs of mice. In the studies reported here, injections of orange oils with low and high lerpene content, d-limo- ncne, and a mixture of d-limonene and 6% of its hydropcroxidc were made into the samc site of C578L/6 lax mice 24 hours aflcr subcutaneous in- jections of 25 r.g DBP in tricaprylin. This procedure reduced the tumor incidence to Icss than 50% after 30 weeks (compared with 80% in con- trols receiving the carcinogen alone or the carcinogen followed by tti- caprylin). 'Ebe time of latency was markedly prolonged. In A/lax mice with spontaneous adenomas and adenomas induced with subcutaneous carcinogen, orange oils, as well as limonene, reduced the incidence of lung adcnnmas when given weekly for from 13 to 16 weeks. The accclcration of tumor growth usually observed following the transfer of multiple prxrlcd carcinogen injectirm silcs into new hosts was inhibited by orange oils IlomAurRrr, F., T reger, A. and Bogcr, Ei. Oncology 2 5:1-1 U, 19 71. Other support: American Cancer Society and U. S. Public Ifcalth Service. From the Bio-Rescarch Inslitute, Cambridge, Maas. I EINFLUSS CIIRONISCHER INHALATIONEN VON FRISCEfEIEM ZIGARETTENRAl1CII IJND DESSEN GASPIIASE Al1F DIE ENTWICKLUNG VON LUNGENTUMOREN BEI SNE'sE.L'S MAUSEN Chronic inhalation of tresh cigarette smoke and of the Fas phase alone contribute to the carcinogenesis of glandular lung tumon in Snell mice. In experiments reported here, one group of mice was exposed chronically, i.e., over the longest possibk time span, to total cigarette smokc, and another group for an equal kngth of time to only the gas phase of the smoke; a third group of Snell mice served as a control. Two t~pes of glandular lung tumon, adcnomas and adenocarcinomas, were observed in these animals. In the controls, the mak animals exhibited twice as many spcx~taneous lung tumors as the females. After inhalation of cigarette smoke the number of lung tumors was increased; this increased frcquency of tumors was particularly noticeable after inhalation of the gas phase. ll+e effect was more marked in mak than in female mice. It is interesting to note that in the controls, most of the tumors were observed only in mice which were nwrc than two years old, whereas in the majority of the ex- perimental mice turnon were found at a much earlier age. BrorKhogenk carcinoma, such as that which frequently occan in human cigarette smok- ers, was never observed in the mice. Since adenomas and adenocarcinomas were found in controls as well as in experimental mice, it seems reasonable to suggest that inhalation of cigarette smoke by mice enhances more or less celrular abnormalities which are already present in the lung of the mice. Leuchtrn6eran, ( : and Leuchtenberger, R. T.riuchri/t fur PravrnrlvmrdrLln 17:457-462, 1970. From the Swiss Institute for Experimental Cancer Research, I.ausannc. 29 28

7C. ~ ...r. ~ .... EFFECTS OF CHRONIC INHALATION OF WHOLE FRESI( CIGARETTE SMOKE AND OF ITS GAS PHASE ON PULMONARY TUMORIGENESIS IN SNELL'S MICE Does chronic inhalation of fresh whoie cigarette smoke conrainin6 particulate matter have the same or a different biological effect than that of inhalation of the gas phase of fresh cigarette smoke? To answer this question. 467 inbred Snell's mice were exposed to chronic inhalatton of puRs of whole frssh cigarette amoke or of its gas phase alone for ont year or longer. The smoking machine used for this experiment permitted ex- posure of individual mice to inhalations of pufls o( cigarette smoke alter- nating with air. Exposure to both whole ciaarette smoke and its gas phase a/one resulted in an earlier occurrence and a higher frequency of gla,.dular pulmonary tumors when compared with those of controls. The enhance- ment of pulmonary tunwri8enesis was particularly striking after exposure to the gas phase alone and was more pronounced in males than In fr males. Males exposed to the gas phase disclosed not only a significant shifr from pulmonary adenomas to ~nocarciramas, but the frequency of adc#xrcar- cinomas was 23%, that is, five times 6reater than that of controls, which was 4 7%. No bronchogenic carcinoma was observed in controls or in mice exposed to whole fresh cigarette smoke ot its gas phase. Leuchttnberan. C. and Leuchtenber8er, R. In Nettesheim, P., Nanna, M. G., )r., and Deatheragc, ). W., Jr. (cds.): Morpholoay of f:xptrinwntol Rtspirorory Corcinotencsis, Oak Ridge. Tenn.: U. S. Atomic Energy Commission, 1970, pp 329-346. From the Swiss Institute for Etperimental Cancer Research, Lausanne. EINFLUSS VON FRISCHEM UGARETTENRAUCH AUF DIE ENTWICKLUNG VON LUNGENTUMOREN UND AUF LUNGENKULTUREN BEI DER SNELL-MAUS TTThis report deals with the efiect of chronic inhalation of cigarette smoke on the developnxnt of lunt tmsors in SoeB's mice, and the influence of citarette smoke on alveolar gn growing out from lung explant cultures of Snell's mice. Results of these e:periments showed that chronic inhalation of whole fresh daarette stnoke, or of ita aas phase alone, occa- sioned Ure earlier occvrreoce and higher frequency of lung tumors in Snell's mice. This enhaneement of pulmonary adenomas and adenocarcinomu was particularly striking after espawre to the 8as phase alone and more pronounced in rnaks than In femaka; no broncho8enic carcinoma was ob- served. Lung explant cultures, which were exposed to whole charcoal- filtered ci8arette smoke fran which cytoloxic factors were nearly all eliminated, showed a aclet.tive damaee to the alveolar macropha8es. After exposure of lung esplant cultures to the 6as phase from ehareo.l-filtered cigarette seake, the alvedar macroPhases showed a aekctive atimulation o( DNA synthcsis. These results indreatt, on one hand, the importance of particulate matter for the Inhibition of cell metabolism, and on the other 30 hand the importance of the gas phase for the stimulat on of cell metabolism of alveolar macrophages. i.euchtenfrerger. C. and Leuchtenberger, R. SchwtiLtrischt MediZinischt Wochtn.rchriJt 101(38):1374-1381, 1971. Other.upport: Association Suisse des Fabricants de Cigarettes. From the Swiss Institute for Experimental Cancer Research, Lausanne. LYMPHOSARCOMA IN THE RABBIT: GENETICS ANI) PATHOLOGY • Lymphosarcomas are rare in rabbits. However, since 1962, 29 rabbits of strain WH at 1 he Jackson Laboratory have exhibited lymphmarcoma. Because of this unusual aggre6ation of lumors, the pedi8rees of the affected animals were analyzed. 'I he inheritance indicated an sutosomal recessive gene conferring susceptibility to lymphosarcoma; this sene was designated Is. The data were compatible with both concepts of genetic susceptibility to lymphosarcoma and vertical transmission of a virus. The neoplastic in- volvement of Iymphoreticular and other orsans, especially kidneys, cor- responded to a pattern observed in lymphosarcoma of other domestic animals. Specificallr, it resembled in many ways visceral lymphos.rtomato- sis of cats which has been proved unequivocally to be caused by feline leukemia virus. Because oC the vast amount of rnorpholo6ic, physiologic, and biochemical data available on rabbits, the simplicity of rabbit care nd breedina, and their large sixe, rabbits are of considerable value in biomedical research. The finding of 1ymphosarcoma and its hereditary basis provides a new nd important model for studies of the pathogenesis of neoplasia. Fox, R. R., Meier, fl., Crary, D. D., Myen, D. D., Norber6, R. F. and Laird, C. W. Journal of the Not{onal Cancer Institute 45(4):719-729, 1970. Ot14er.rpportr U. S. Public Health Service and Hycet, Inc., Houston. From The Jackson Laboratory, Bar Harbor, Me. MALIGNANT TRANSFORMATION INDUCED BY 7,12- DIMETIiIYLBENZ(a)ANTHRACENE IN RAT EMBRYO CELLS INFECTED WfTH RAUSCHER LEUKEMIA VIRUS Morphological alterations of cells and an abnormal growth pattern were noted 42-45 days after trestment of Rauscher kukemia virus ( R1.V ) infected rat embryo ( RE ) cells with various levels of 7,12-dimcthylbcnt- (a)anthraeene (UMBA) for seven days. Uninfected cells treated with DMBA and RI.V-infected RE cells untreated with DMBA failed to show any evidence of transformation. When stained with Giemsa, the foci of transformed cells consisted of randomly oriented eriss-cn.csinR spindle- shaped cells having much more rapid replication rates than the untreated and untransformed RE cells. The transformed cells were more resistant 31

m ~ r m to the toxicity of DMBA than were the untreated RE cells and aily the trancformcd cells were capable of producing local subcutancous sa •cornas when transplanted into newborn rats. When re-cstahlished in tissuc (ulture, cells derived from the tumors, like Ihe tumor tissue itself, contained group- specific complement-fixing antigens characteristic of the murine Icutcmia- sarcoma virus complex nd the C-type RNA particles. These results, which showed that both chemical and virus were required for transforriation, suggest that the C-type RNA viral genorne of RLV provided !pccific oncogene information for the malignant transformation. Rhim, J. S., Vass, W., Cho. I{. Y. nd Nucbner, R. 1. (Microhiologicof .1 rsuciarcr. Inc.) International lournal of Cancer 7:65-74, 1971. Ofhrr anpporf r National Cancer Institute. From the Department of Virus Rcsearch, MicrobiolrsRical Assr.ciata, Inc., and Viral CarcirnoRcrsesis Branch, National Canccr Institute, Bcthc.d.i, Md. POLMONITE INTERSTIZIALE CRONICA IN CORSO DI SARCOMA MURINO DA METILCOLANTRENE Ilistological examination of the lungs of 147 mice with mcthylcho- lan(hrcne-induced sarcomas or transplants of the same tunxxs revealed 24.5% chronic interstrtial pncumonia. Mice used for this study were of the C3H,B/Cb/Se. R111/I)m/Se, CBA/Cb/Sc, nd FI(C3f1.8/(_'h/Sc X RI11/Dm/Sc) strairn The pneumonia seen in these mice differed from bronchopncumonia, which is frequently scen in old micc, by reason of the age of onset, the ahscnce of alveolar exudatc, and the intense proliferation of septal cells. Malt7efl, N. (Severi. 1, / Lav. Anar. Pat. Prrugia 30( 1):1 S-2S, 1970. From the Division of Cancer Research, University of Pcrugia. Italy. I TUMORI I)El- POLMONE. DEL RETICOLO E DEI.LA MAM- MELLA NEI TOPI BALB/c/Cb/Se DISCENDENTI DA GENITORI TRATTATI CON URETANO NEL PERIOE>D NEONATALE: Two groups of BALB/c mice were treated as newborns with one and five milligrams of urethane respectively, and then mated. Offspring of these treated mice were followed in a study of the incidence of spontancous tumors. Descendants showed an increase of lung and reticular tunxirs, in male and female, virgin and mated mice. A discussion of the mcchanism through which urethane, inoculated into newborn mice, enhances spon- taneous tumorigencsis in the next generation is rcpcxtcd. Ribacchi, R. (Severi. L.) lav, Anor. Par. Prrulia 30( I):27-39, 1970. Fmm tlrc INvitii<,n 44 ('ancer Rexarch, University of I'crugia, Italy. 32 DF.TEC-IION OF A LUNG TUMOR AGENT IN IIALB/c/(b/Se IRRADIA'f E:D MICE In this inve-,tigation lung tissue from newborn mice was graflcd into adult syngcncic recipients previously panirradiated in an attempt to meet the most suitable cxpcrimcntal conditions for the detection of cndogenous lung tumor agents. Of the 29 recipients irradiated before grafting. 21 had recoverable lung grafts and five of thcm, or 23.8"6, showed alveolar adcnomas in their lung grafts. The number of lung grafts recovered from irradiated recipients was 44. No tumors were observed in the 43 grafts of newborn lungs recovered from 23 non-irradiated control recipients. A strong association was obscrved between leukemia in irradiated recipients and adenomas in the lung &rafts. Although four of the irradiated recipients died spontaneously of thymic leukemia and the recovery of lung grafts was not possible, four other mice were kukemic when killcd and three of these had recoverable lung grafts showing alveolar adcnomas. 'llasc results suggest that whole txxiy irradiation activatcs a lung tumor agent in BALB/c mice. Squartini. F. and Bolis, G. B. (Severi. L.) Lav. Anar. Par. Prrugia 30( 3):125-128, 1970. From the Division of Cancer Research. University of Perugia. Italy. DIVERSA SUSCF.'fTIBILITA DEGLI EPITELI A(-VEOI.ARE E TRACNEOBRONCEf1ALE DEL TOPO BALB/c/Cb/Se ALLA TUMORIGENESI DA URETANO NE:1 TRAPIANTI ISOGENICI Data relevant to the susceptibility of alveolar and tracheobronchial mouse epithclium to urethane tumorigencsis are reported. Grafts of trachea together with the lower section of larynx, of main bronchi, and of peripheral lung tissue of newborn BALB/c mice were subcutaneously impIanted in 2-moxtth-old isogenic mice. Five days after grafting, one group of mice was treated intraperitoncally with a single injection of urethane at the drne of I mg/g of body weight. Another group of untreated mice was used as thc control. Six months latcr, 86.4% of femaks and 96.1 % of males developed lung tumors of alveolar origin in the grafted lungs. No hyperplaslic, prcncoplastic, and/or neoplastic Icsion caused by the carcinogen was observed in the trachcal, laryngeal, and bronchial grafts No tunnors were obscrvcd in the controls. Ribacchi. R. (Severi. 1..) Lav. Anat. Pat. Prrumia 30(3):147-160, 1970. From the Division of ('anccr Rescarch, University of 1'crugia, Italy. 33 U CA A r

n ....rl 76, ...3. ~ 111. Crrrdiorasr•ulrrr System TH1: EVALUATION AND TREAI MF.NT OF C'ARDI(Xil'.NIC SHOCK This thorcwgh review paper discusses the clinical assessmcnt and therapy of cardiogenic shock and presents new information which coutd mcxhly n overall approach to the use of vasopressor agents in shocF. For diagnostic purpincs, shock is conventionally classificd into hypovolcmic (hcmorrhagic), scptic, and cardioglt:nic types. In each of these shock classifications, early diagnosis and institution of treatment are the l cy to improved survival. An cight-point, annotated schedule for Ihe assessment of the shock state is presented here. In cardiogenic shock, the goal of therapy is the restoration of myocardial function to the point where it can adequately provide the contractik force necesttary for the perfusion -4 the systemic circulatirx+. 'il+e use, as well as choice, cd vasoactive aRcrts for maintenance of satisfactory arterial prcuure in cardiogenic .:hock is u very controversial subject right now. Vasoactive agents discussed in dctai, here include I-norepinephrinc, isoproterenol, chlorprnmazine, mcterarninol, mctham hctamme, and dopamine; steroids and antibiotics also arc con- sidered ~rKny. Mucllcr, I(. S., Ayrri, S. M., Gregory, 1. 1., Giannclli, S., Ir., and (',race. W. 1. Mrdicol Timrt 98(7):137-152, 1970. Oth.r support: (1. S Public Hcalth Service. From the Departments of Medicine and Surgery. St. Vincent's Ilospitr:l and New York University School of Medicine. New York City. THE LUNG IN SHOCK: ALVEOLAR-CAPILI-ARY GAS EXCHANGE IN THE SNOCK SYNDROME In an attempt to delineate the effects of circulatory collapse on pulfnonary ventilation, this paper reviews certain physiologic considerations relevant to abnormal alveolar gas exchange in the patient with shock. Alveolar gas exchange in eight patients with shock revealed marked reduc- tion in arterial oxygen tension (PaO2) and increase in the alveolar-artcrial oxygen difference while they were breathing room air. Failure to adequately increase PaOt while breathrn6 100% oxygen indicated significant increases in venoarterial shunting nd demonstrated persistent perfusion of non- ventilated lung units. An Increase in the alveolar-arterial carbon dioxide difference indicated increascd alveolar dead space, suggesting that some alveolar units were undet'ptrfused. Continuous pressure breathing improved PaOn in six of seven patients but tendcd to incrcasc venous prescurc and reduce aortic pressurc. These obserraticxn suggest that alterations in lung functicxr are part of the generalized metabolic dccnmpencatiim seen in shixk I)ccrcascd pulmonary nutriticxr after decreased hkKK1 flow, incrcascd 34 Icft atrial preccurc, and pulmonary venoconstriction arc all implicatcd. Progressive pulmonary changes may preclude revcrsihility t+r thc shock state. Ayres. S. M., Mucllcr, 11., Giannelli, S., Jr., Fleming. P. and Ciracc. W. 1. The .1 mcricon lnurnul of Cardiology 26( 6): 588-594, a 970. Other support: U. S. Public Health Service and New York Ilcart Association. From the Departments of Medicine and Surgery, St. Vincent's liospital and Medical Center of Ncw York and Ncw York University School of Medicine, New York City. MYCX'ARI)IAL AND SYSTEMIC RESPONSES TO CARBOXYIII:M(XiLOBIN ' This paper reviews ceblain theoretical aipecls of systemic and myo- cardial oxygen exchange and summarizes a series of experimental atudies of the physiological eRects of relatively small amounts of carboxyhcmo- globin (COIIB). Measurements of alveolar-capillary gas exchangc, sys- temic hemodynamics, and myocardial metabolism were made in a series of h.tman and canine studies before and after the administration of amounts of either 5% or ().I % carbon monoxide sufficient to raise COILB satura- tion to between S and 25%. Arterial and mixed vcnous oxygen tcnsirms were decreased by administration of either concentration and changes could be identified with COHB saturations below 5%. Cardiac output and minute ventilation increased when COHB was elevated by breathing the higher concentration. The coronary arteriovenous oxygen difference was uniformly decreased and coronary blood flow increased when COIIB was raised to between 5 and 10% saturation with either gas mixturc in the human studies. Certain patients with coronary artery disease developed altered lactate and pyrvvate melabolism following elevation of COIIB suggesting myocardial hypoxia. Significant myocardial changes were seen in patients with ckvation of CO11B above 6% COHB. The caninc prcpara- tion, in contrast, was notably moro rrsistant to COHB and significant al- terations were not observed until COIIB saturation exceeded 25%. Ayrrs, S. M., Giannelli, S., Jr., and Mueller, H. Annals of the New York Academy of Sciences 174 ( 1): 263-293, 1970. Other .rpport: National Air Pollution Control Administration and Na- tional Heart Institute. From the Dcpartments of Medicine and Surgery, St. Vincent's 1{ospital and Medical Center, New York City. FIEMODYNAMICS, CORONARY BLOOD F1AW, ANI) MYOCARDIAL ME:TABOI-ISM IN CORONARY SLI(X'K; RESPONSE TO /-NOREPINEPHRINE AND ISl)PROTI:RI:NO1. This paper prescnls myocardial mctalxrlism and systemic hcrrw>dy- 35

namic data in IR patients with severe coronary shock. Cardiac index (CI) and stroke index were markedly reduced in all patients. Mean arterial pres- sure ranr.ed from 40 to 65 mm 1(g while s~stemic vascular resistancc varied widcly. ('oronary blocxl flow (C'BF) was decreased in all but three paticnts. Whitc myocardial oxygen consumption (MV„) was normal or incrcased, myocardial oxygen extraction was above 70% and coronary sinus oxygen tension was below 22 mm Hg in most of the patients. The detection of the abnormal oxygen pattern in tpite of sampling of mixed coronary venous blood indicates the severity of myocard'ul hypoxia. In 15 patients myo- cardial lactate producticxt was demonstrated; in the remaining three lactate extraction was below 10%. The response to f-norepinephrinc was studied in seven cases and the response to isolxotercnd in four. Considcrition of both hemodynamic and lactate flux data suggests that I-norepinephrine is supericx to isoproterznof In corortarr sfack. F.atly administration o' dilute u~ution of f~ncxepirxphrine, carcfully regulated hr intra•artcrial pressure monitoring to prevent excessive increases In ventricular work, rnig it well preserve the integrity of noninfarcted rcgsorts o! thc myocardium and prevent the devcbpment of circulatory collapse. Mueller. Ff., Ayrrs, S. M., Grrgory,l. I., Giannclli, S., 1r., and Gracc, W. 1. The Journal o/ Clirrko! Invrstlrarion 49(10):1885-1902, 1970. Other rrpportt National Heart Institute. Fmm the Departments of Medicine and Surgery. St. Vincent's Ftospital and Medical Center, New York City. ALTERATIONS IN 2-3 DIPHOSPFIOGLYCERATE AND 0. HEMOGLOBIN AFFtNITY IN PATIENTS UNDERGOING OPEN-HEART SURGERY Since it has been shown that hemogiobin affinity for oxygen is inversely related to the kvel of red blood cell (RHC) 2-3 d'iphosphoglycerate (2-3 DPG), both of thesepa nmeten were measured in 14 patients undergoing heart-lung bypass to define the influence o( this procedure on 0. deliver7 in patients undergoing open-heart surgery. RBC 2-3 DPG decreased with institution of bypass and continued to decrease during bypass. The day following heart-lung bypast RBC 2-3 DPO kvels returned to normal, but were lower than preopcrative control values. In vitro studies denxmstrated that a portion of the ItBC 2-3 DPG loa was due to circulation of blood through the oxygenator itsclf, but other factors also appear to be involved. Hcmo- globin affinity for oxy¢n increased durfng heart-lung bypass in most patients. However, It was found that the incrcased affinity was less than would be expected relative to the demonstrated RBC 2-3 DP(7 decrease, and that a progressive decrease In RBC pH partially corrected the Itera- tion in 0, aflinity. The data suggest that other factors such as carbon nx,n- oxick levels nd rate of RB(' destruction by the reticuloendothclial system are involved in the demonstrated alteration in hemoglobin affinity for 0,. ~ 36 1 r a, Bordiuk, 1. M., McKcnna. P. 1., Giannclli, S., Jr., and Ayres, S. M. Supplement I to Circulariun XLIII and XLIV:1-141-1-146, 1971. Othrr.upport: U. S. Public Hcalth Service. From the lkpartmcnts of Pcdiatrits, Pathology, Surgery and Medicine, St. Vincent's )Iospit:d and Medical Center, and New York University School of Mcdicinc, Ncw York City. TOTAL AND NUTRITIONAL CORONARY FLOW The first part of this methodology paper describes a new technique for determining the velocity of red cells in the capillaries. Such rrxasure- mcnt is accomplished by r:etermining the frame to frame progress of iodi- vidual red cells in movinj ~turn of the capillaries. So far data have denwmstratcd that rnrrcpincpbrine kads to the opening o( capillaries, that nitroglycerine slows the flow in the capillarks, and that methachotinc re- sults in the complete arrest and even retrograde flow. The remainder of the report is concerned with the measurement of total and nutritional coronary ftow by means of coincidence counting with a positron emitter, rubidium-84. Nutritional as well as totaL coronary flow can be determined by means of the ckarance or the Fick 2rincipk, respectively. Results of this technique demonstrate a dichotomy of the coronarr crrculation and illustrate that the action of drugs on the coronary circulation can vary with respect to nutri- tional and total flow. Norepinephrine causes a disproportionately greater increase in total as compared with effective Oow, and lhe same is true for nicotine. Isoprotercnol, on the other hand, causes a proportionate increase in both the nutritional and effective coronary flows. Tillich, G., Mendon, L. and Bing, R. J. Circulotion Rtsrarch XXVIII and XXIX (suppi 1):1-148-1-153, 1971. Other .upport: U. S. Public Health Service and Hartford Foundation. Frc,m the Huntington Memorial Hospital, Pasadena, Cal., and the Uni- versity of Southern California School of Medicine, Los Angeles. THE EFFECT OF INTRAVENOUS ETHYL ALCOHOL ON THE CORONARY CIRCULATION AND MYOCARDIAL CONTRA(TILITY OF THE HUMAN AND CANINE IIEART In this attempt to examine closely an area of some experimcntal con- troveny, the effects of intravenously administered ethyl alcohol on coronary blood flow, contractility, and hernodrnamks were investigated in 10 pa- ticnts and in 23 anesthetized dogs. Results obtained In man with a dose of alcohol comParabk to that contained In two to three ounces of whiskey showed a consistent but not significant increase In effective coronary flow accompanied by a fall in coronary vascular resistance which was also not significant. There was no significant change in cardiac index, heart rate, or peripheral vascular resistance. Likewise, the studies conducted with dogs showed that when small amounts of alcohol were infused the effective 37

coronary flow rate diminished slightly but not sipiificantly, without signifi- cant changes in coronary vascular resistance. Ffowever, when the dcnaRe levels were increased, a significant increase in effective coronary flow was noted along with a significant fall in coronary vascular resistance At a mean alcohol blood level of 195 mg/I()0 cc of bkood, there was a marked diminuticxt in the velocity of left ventricular contraction; the reasons for this diminished myocardial contractility are not clear. Mendoza, L. C., Ifellberg, K., Rickart, A., Tillich, G. and HinR. R. l. Journal of Clinical Pharmacology 11( 3):165-176, 1971. Ot/4rr sopprt: U. S. Public Health Service. F rom the Huntington Memorial Hospital, Pasadena, Cal , and the (1ni- vcrsity of Southern California School of Medicine, I,os Angeles. FFFE:CT OF NITRCX7LYCERIN ON TOTAL AND Rf?(il()NA1. ('ORONARY Bl.(X)D FLOW IN THE NORMAL ANI) IS('IiE?MIC CANINE MYOCARDIUM In experiments rcported here, fractional extraction of Rb-86 was used to estimate the regional distribution of coronary flow in adult mongrcl dogs. With this method, the effect of nitroglycerin on total coronary fbw and its regional distribution was determined in normal conditions and in myo- cardial ischemia. In normal hearts, capillary flow was 9.5% higher in the inner than in the outer myocardial half. When partial occlusion of coronary arteries was followed by infusion of noradrenalinc, subendocardial flow was less than capillary fbw in subepicardial layers. In these conditions the administration of nitroglycerin resulted in a redistribution of capillary flow towards normal. Mathes. P. and Rival, 1. (Ring, R. !.) Cardiovascr.lar Research S( I):54-61, 1971. Oth.r arppori: Michigan Heart Association, American Medical Associa- tion - Education and Research Foundation, and Detroit General Ilospital ReWarch Corporation. From the Wayne State University School of Medicine, Detroit. TIIE ESTIMATION OF BLOOD PLATELET SURVIVAL: II. THE MULTIPLE HIT METHOD The controversy over the pattern of platekt destruction has not yet been resolved. Some contend that it is time-hornogcrxous and inctiscrim- inate. However, a great many studies have bcen published in which the pattern of decayboth in man and in experimental anrmals, conforms well to "linear" decay pattern over a period of 10 days or so. This paper develops four mcukls Eor estimation of blood platckt survival. llrey are presented not as compktely rcalistk but as at kast first order approxi- mations which shxwld at once providc a systematic basis for estimntion of mean platelet survival and the standard error of the estimate. 'They will at the same time circumvent the subjective clement in analyzing the data with the attendant biases and also provide a unifying theory for the several different models which various investigators have more or less explicitly used in the past. 7 he models may perhaps be thought of as smrwrthing functions to iron out the high experimental error inherent in the study of platelet survival. Primarily, an attempt has been made here to construct the model on the basis of present knowledge of the economy of the platelet and the factors influencing its survival. Murphy, E. A. and Francis, M. E. Thrombost,r et Diatherit Iltmorrhoaicn 2S(1) : 53-80, 1971. Othrr artport: U. S. Public Health Service. From the Departments of Medicine and Biostatistics, The luhns Ilupkins University Schcx>f of Medicine, B.Itimore. ARRHYTHMIAS FOLLOWING INFUSIONS OF FATTY ACII>S In experiments reported here, 10m1/k& of a I in 1.000 w/v suspen- sion were infused into dop over a five-minute period. The acids infused were palmitic, stearic, okx, linofenk, and combinations of oleic-palmitic and okic-stearic. A total of 178 dogs were used in these studies. A char- acteristic sequence of ekctrocardiographic abnotrnalitiea occurred after the rapid infusion of long-chained saturated fatty acids (stearic, palmitic). In addition to an injury current, elcctrocardioBaphic changes were sinus bradycardia, supraventricu:ar bradycardia, varying degrees of intraventric- ular block, sinus standstill, and complete standstill. Similar infusions o( monounsaturated (oleic) and polyunsaturated (Iinoleic) acids and a slower infusion of large quantities of long-chain saturated fatty acida given for hours day after day produced no untoward clinical or electrocardiographic effects. The differences in effects between the saturated and unsaturated fatty acids and the differences in effects of rapid and slow infusion of saturated fatty acids might be due to differences in the rate of binding of these acids to albumin. Solo#, L. A. America Hrarr Journal 80 ( S): 67 I-674, 1970. OtlLer arrport: U. S. Public Health Service. From the Division of Cardiology. Temple University Health Sciences Center. Philadelphia. I SIMULATION OF "LEFT ATRIAL RNYTHM" BY RIGIIT ATRIAL PACING This case history report acxrcerns  patient with ( I ) the criteria for the diagnosis of so-called kft atrial rhythm and (2) a similar pattern produced by a catheter pacemaker stimulating the wall of the mid-right strium, low right strium, and coronary sinus. Results rwNed, ahM,uRh not negating a left atrial site for pacemaker activity, suggest a mcrrc likely .ite 39 39

i in the nodal or A-V I'unctional area. The findings presented here serve as evidence for the need of increased caution before assuming that a left to right pattern of atrial depolarization, as interpreted in leads I, V„ and V, of the scaler electrocardiogram, can pinpoint the anatomic origin of the pacemaker. Rutenberg, If. L. and Solo#, L. A. The American Journal of Cardiology 26( 4):427-431, 1970. (htirr arpport: U. S. Public Health Service. From the Division of Cardiology, Temple University Health Sciences Cen- ter. Philadelphia. INFt.UENC'F. c)F ANTIMACiULANTS ANt) AtrrONOMIC DRUGS ON PLASMA FREE FATTY AC'lM ANf) (:LU(Y)Sf: The initial level of free fatty .cid ( FFA ) in the dog may be a critical factor in determining whether or not anticoagulants will csert significant lipol~tic .ctivit~. in csperiments rcported here, anesthetized dogs, which had been hsted overnight, received knrn quantities of heparin or sodium polyanhydromannuronic acid sulfate (Manuronate) in dilute solutions. In animals so fasted, initial pre-drug plasma FFA kvets were low and quite comparable and no significant changes in FFA or glucose occurred after admmistration o( hcparin or Manuronate alone. In other groups similarly fasted and receiving hep.rin, the intravenous infusion of nicotirx, epi- nephrine, or isoprotercnol induced clearly significant elevations in plasma FFA akmg with increases in plasma glucose. Non-fasted dogs, cxr the other hand, showed high initial kvcls o( plasma FFA that were further increased with great variability by hcparin a(one while glucose kvels were rclatively unaffected. Tlxse results indicate that, under the experimental conditions described, valid experiments can be performed in dogs in the presence of heparin or Manuronate while studying the influence on plasma FFA of certain other compounds such as autonomic agents. PaPaeostas, C. A., Weiss, L. M. and Soloff, L. A. Archives /nrrrnarionalrt de Pharmocodynomit et de Thfrapir 187( 1): 46-51, 1970. Other arpport: U. S. Public Health Service nd National Ifeart Institute. From the Departments of Pharmacology and Cardiology, Temple Univcr- sity Medical Center. Philadelphia. POSSIBLE MECNANISM OF EGRF.SS OF FREE CHOLESTEROL FROM THF: ARTERIAL WALL If n equilibrium enists betwettt free choksterol of the serum and of the arterial trs.ue, the rate of esterifkatioo of free chc>tcsterul mirht he an important factcx in alk>..ing more cholesterol to leave the arterial wall. To test those hypothesis, segments of human iliac arteries (treated and un- treated) were incubated with human serum (treated and untreated). In the ca%e of both arteries and serum. treatment consisted of heating the sample at S6' C for 3t/ minutes to inactivate the enzyme lecithin-choiesterol-acyl- transfcrase (LCAT). When LCAT enzyme was present in the incubation medium, virtually only free cholesterol kft the arterial wall, whereas when LCAT enzyme was absent, only cholesterol ester left the wall. These re- sutas suggest that a decrease in serum frtt choksterof, caused by a serum cholesterol estcrifymg enzyme, pouibfy LCAT, might be important in promoting the removal of free cholesterol from the arterial wall. Since cholesterol esterifxation has been shown recently to be significantly im- paired in subjects with acute myocardial infaretion and chronic coronary arterr disease. it appean that a deficiency of cholesterol esterifying enryme, possiMy LCAT, may be important in the etiology or rate of development of athero.clerowis. Rutenberg, It. I.. and Soloff, L. A. Nature 230: 123-123, 1971. Other auppert: U. S. Public Health Service. From the Department of Medicine, Temple University Flealth Sciences Center, Philadelphia. STUDIES ON FIGURE DRAWINGS: BIOLOGICAL IMPLICATIONS OF STRUCTURAL AND GRAPHIC CNARACTERISTICS The Figure-Drawing Test has been used since 1951 in the Study of the Precursors of Hypertension and Coronary Diaease to see whether figures drawn in young adulthood by subjects who develop hypertension or cor- onary disease in middk life are significantly different from those of subjects who remain free from these disorders, and to identify the nature of the differences. Figure drawings by 837 lohns Hopkins medical students in the classes of 1952-1964 are included in the present analysis. The bulk of the drawings were by 725 white male students in the classes of 1952-1963; drawings by 50 white women students in these classes were used for the sex comparison. Drawings by 62 white male subjecU in the class of 1964 were added to the original male group for comparisons involving physic~ logical and metabolic variables. The results of this study indicate that the structural and graphic characteristics of figure drawings may indeed have biological corrclates, so that individuals of different makeup tend to draw different kinds of figures. For exampte, men draw the male figure first more often than do women and thinptop k draw figurrs with bclts more often than do fat people. In view of the fairly extensive literature on per- sonality differences between smokers and rronsmoken, it is purzling that their figure drawings showed ao few differences. The final significance of the study's findings remains to be ascertained in view rH' the hyputhesis-sctking nature of this investigation. 40 41

Thonras, C. R., lones, L. W. and Ross, f). C. Psychiorric Quartrrly Supplrmrnt 42( 2):223-251, 196R. Other .rpport: National Neart lnstitute, National Institutes of llcaltti and International Business Machines Corporation. from the Departnxnt of Mcdicine. The Johns Hopkins University School of Mcdicine, HalNmore. IV. Respiratory System EFFECT OF CADMIUM TON ON RESPIRATION AND Al Pase A('TlVITY OF THE PULMONARY ALVEOLAR MA('ROPIIi%GF?: A MODEL FOR TIIE STUDY OF F.NVIRONMF.NTA1. IN f!?RFERl:N('!: WITH PULMONARY CF.1.L F(1N(-I ION I'ulmonary alverlar mactophages (PAM cells) were isolated from sheep and respiration studies were conducted at two levels of organiz.tion: intact PAM cells and iaolated mitochondria. App roximately SmM "d=# inhibited cndogenc.us respiration in PAM cells and 0.5 mM Cd24 inhbited respiration in the isolated mitochondria. Tetramethyl-p-phenylenediamine, an artificial electron carrier, restored the Cd=4 -inhibited respiration in both cells and mitnchondria, indicating that Cd24 presumably bound certain respiratory chain comporrcn,s. Cadmium ion also inhibited the Na t-K r, Mg=t ATPasc syttcm of PAM cells and cellular membranes. The results of this investigation have quantitated in virro the inhibitory effects of Cd' t, a constituent of both urban cnvironments and cigarette smoke, on cellular and subcellular biochemKal pmcesses of the PAM cell. Although separate studies are necessary to correlate these observations with Cd=+-induced pulmonary diseascs, the model presented here affords an opportunity (or studying in virro an isolated and important lung cell system and assessing the effects of environmental agents potentially hazardous to pulmonary tissue. Crost, C. E.. Ibrahim, A. B., Ahmed, M. and Mustafa, M. G. Environnunrol Rr.rrorch 3(S-6):512-520, 1970. Other .upprt: Tuberculosis and Respintory Disease Association of California. From the Department of Internal Medicine, University of California School of Medicine, Davis. INTERFERENCE OF CADMIUM iON WITFI OXIDATIVE METABOLISM OF ALVEOLAR MACROPIIAGES Most of the enerEy-requirlna processes of pulmonary alveplar macro- phages are dependent upon beirohic metabolism. !n vitro studies show that these cells consume 0.15 b 0.2 rmolar 0s/mg protein/sec. Mito- chondria from alvrdar macrophages manifest a respiratory rate of 0.5 to 06 rmolar 0./m` protcin/aec for succinate as a substrate. Mitochondrial oxidation is coupled to phosphorylation; adenosine diphosphate (ADP): oxygen ratios are approximately 2 for flavin-linked and 3 for pyridine nuclcotidc-linked substrates. Cadmium ion, a common ir pollutant and a reported component of cigarette smoke, adversely affects respiration of alveolar macrc.phages. It completely inhibits macrophage mitochondrial oxygen uptake at 50 rmolar concentrations and uncouples oxidative phos- phorylation at 5 Nmolar concentrations. C::d+ +nd several other divalent cations, like Cu r+, Hg 4 +, Sri+ } and Zn++, also inhibit adenosine tri- phcnphatase ( ATPase ) activity of alveolar macrophages. Since metal ~umes and oxides are common air pollutanls, a study of this sort might provide information on mechanisms at the biochemical kvcl as to how toxic inhalants initiate pulmonary pathology. Mustafa, M. G., ('rotr, C. E. and Tyler. W. S. ArchivtJ of Inrrrnuf AlydkJnt 127(6):1050-10SS, 1971. Other support: (l. S. Public Ilealth Service and California Tubcrcuk»is nd Respiratory Discasc Association. From the Departments of Biologjcal Chemistry. lluman Physiology, ln- tcrnal Medicine, and Anatomy. University of California School of Medicine and Veterinary Medicine, Lhvis. TlIE ORIGIN OF PFJLMONARY ALVEOLAR MACROPHAGFS: STUDIES OF STliM CELLS USING THE Es-2 MARKER OF MICE Although it is well known that pulmonary alveolar macrophages (PAMs) constitute a major lung cellular defense mechanism. there is at this time a considerable controversy concerning the cellular precursun of the resident PAM population. In this attempt to identify the prccurxrr, genetically related IRF/Al(+) and RF/AI(-)) substrains of mice were used to investigate the possibility o( PAM origin from marrow stem cells. The RF/Al (-4- ) mice carry a notn(xci8c readily identifiable prealbumin esterase marker, Es-2. Reciprvcal chtmerism was produced by administer- ing a lethal dosc of a-irradiation to (() RF/AI(-) mice and subse- quently injecting them with RF/AI( +) bone marrvw, and (2) RF/AI (-}- ) mice and subsequently injecting them with RF/AI(-) bone marrow. Eight weeks after irradiation pulmonary macrophagra were obtained by bronchial lavage. PAMs from RF/Al(-) animals transplanted with RF/AI( -{- ) bone marrow showed the prealbumin esterase marker. Es-2, while PAMs frnm the RF/Al(+) animals given RF/AI(-) bone marrow had all but lost their Es-2 marker band, which oonfirau the marrow origin of PAMs. Brunstetter, M-A., llard;e, J. A., SchiR, R., Lcwls, 1. P. and Cross, C. F_. Archives of Internal Mrdicint 127(6):1064-196g, 1971. Other .upportr (l. S. Public Health Service and Atomic FnerRy ('(+rn- mission. From the Department of Medicine. University of California Schrwrl (d Medicine. Davis, and the Department of Anatomy, lbfts University Schtrc)l of Medicine, Boston. 42 43

PULMONARY ALVEOLAR MACROPHAGE: MEMBRANE ASSOCIATED SODIUM ION, POTASSIUM ION, AND MAGNESIUM ION ADENOSINE TRIPHOSPHATASF. SYSTFM This paper prescnts one type of cellular ATPase system, a system which appears to be located prrmarily on the plasma membranc, to be activated by Na+ nd K+ and specifically inhibited by the cardiac gly- coside, ouabain. According)y, this enzyme srstem is presumably a part of a transport "pump mechanism of the pulmonary alveolar macrophage (PAM) cell memkrnne. The characteristxs of this ATPasc system were investigated in subcellulv fractions and in intact cells derived from rabbits and sheep. Equimodar amounts of M6++ and ATP were needed for rnaxi- mal activation by Nat or K+. Whik Na+ or K+ alone had a Iittk cflect on ATPase, simultaneous additioe of both catiorn produced a 50% atimufation of basal activity. The Nar-K• M4+4ATPue activity was inhihited by ousbain but was inscrritive to ohjomycin and dinitrophenol. I:Lrthcr studies revealed that this Al Paae activity was located predominantly in the plasma membrane and ecnerated 20rmoh Inorganic phosphate per milli- sram of protein per hour at 30' C. This membrane-associated Al'Pase may be interrelated with cellular enersy-rtquiring processes, which oier a biochemical measurement suitable for study of the interaction of air pol- lutants with the cell membrane of alveolar macrophages. Cross. C. F.. Mustafa, M. G., Peterson, P. and Hardie, J. A. Arrhivts of Internal Medicine 127 ( 6): 10fi9-1077, 1971. Other arpportr California Tuberculosis and Respiratory Disease Asscxia- tion. From the Departments of Internal Medicine and Biological Chemistry, University of California School of Medicine, Davis. LOCALIZATION OF NA +-K+ M¢+ + ADENOSINE TRIPIIOSPIIATASE ACTIVITY IN PULMONARY ALVEOLAR MACROPHAGE SUBCELLULAR FRACTIONS •Pulrnonary alveolar macrvphayes possess an enzyme system. Na+-K+ Mgt + ATPase, that may be related to actrve cation transport and inward bulk flow mechanisrro. For the present studies, pulmonary alveolar macro- phages from sheep were collected, fractionated, and analyzed for protein nd ATPase activity in the various cell fRactions. One membrane fraction, namely the middle layer from a density gradient spin, contained most of the macrophage cellular plasma membrane and exhibited the highest ATPase activity compared to other membrane fractions. Ouabain inhibited ap- proximately 30% of Na+-K• Mgf *ATPase activity in both intact cells and membrane preparationa. This inhibition by ouabain implies that this ATPase wbserves active cation transport across the alveolar macrophage plasma membrane. Since Na r-K+,Mg* 4 ATPase was sensitive to ouabain and p-chloromercuribenzoate but insensitive to oligomycin, it seems that this enzyme system nray be related to energy-dependent mcmbranc-linkcd proccucs o( alvcolar macrophages. Mustafa, M. G., Cro5s, C. E. and Hardie, J. A. Life Sciences 9( 1):947-954, 1970. Other aupport: Tuberculosis and Respiratory Disease Association of California. From the Departments of 13iological Chemistry and Internal Medicine, Uni- versity of California School of Medicine, Davis. EFFECTS OF DIVALENT METAL IONS ON ALVEOLAR MACROPHAGE MEMBRANE ADENOSINE TRIPHOSPIIATASE ACTI V ITY This study of the basic mechanisms of patholo~ic change in the lung shows that a number of divalent metal ions exert inhibitory effects on the adenosine triphcnphatase (f,TPase) activity ofpu Imonary alveolar macro- phage ( PAM ) cells. "f he o•,crall survey showed Ihat the inhibitrxy effects of metal ions were variable. Cd+ +, Co++, CuI +, Fe4 4, 1f6t +, Pb4 + Sn++, Sr++, and 2nF + in'tibited the ATPase system by 30% or higher t 0.5 to 2.0 mM coneentra(ions. Other ions such as Ba+ +, Ca+ +, Mn ++ and Ni++ were less inhibitory. Since cadmium (which is both a common air Pollutant and a component of cigarette smoke) can induce both pu1- monary emphysema and Mlmonary fibrosis In man, the effects of Cd ++ were studied in greater detail. The endogenous oxygen uptake of PAMs was almost completely inhibited in the presence of 5 mM Cd r+. A 50 MM concentration of Cd+ • completely blocked the oxidation of succinate nd a-oxoglutarate, and a 5 rM concentration of Cd++ abolished the phcn- phorylation and respiratory control in PAM mitochoadria. On the grounds of these results, a relationship between divaknt cation-induced injury of PAM biochemical machinery and the deleterious effects of certain divalent cations on pulmonary tissue is suggested. Mustafa, M. G., Cross, C. E., Munn, R.1. and Hardie, J. A. The Journal of Laboratory and Clinical Medicine 77 ( 4): 563-571, 1971. Other arpport: Tubercuknis and Respiratory Disease Association of California and U. S. Public Health Service. From the Departments of Biological Cfxmistr7 and Internal Medicinc, tlni- versity of California School o( Mcdicine, Davis. I DETERMINATION OF ALVEOLAR SURFACE AREA ANl) TENSION FROM IN SITU PRESSURE-VOLUME DATA This investigation set out first to determine the total alveolar surface area of the cat lung from the In situ pressure-volume ( PV ) curves for both deflation and inflation, and secondly to determine the alveolar surface ten- sion as a function of the alveolar surface area for the complete cycle from the same in situ gas and liquid PV data. To accomplish these ends cats were ncsthetized and placed, with the chestopc ned, inside a whok•body pkthysmograph. In recording gas and liquid PV eurves, 100% oxyFcn and Ringer-Lcxke solution were used. fias and liquid flows were varied from 44 45

~ ~~ f~J ...1. . 2.9•6.0 ml/sec and 0.S-S.0 mI/sec respectively, with no change in either the hysteresis nr the shape of the PV curve. A constant, K. was determined by comparing the gas and liquid inflation curves at maximal lung volumes and using the equation K- 3PVV5/2y, where y was assumed to bc 50 dyne/cm. The mean value for K in eight experiments was 1225 -!- 100 SE:M. At a functional residual capacity of 90 ml the alveolar surface arca of the cat/kR body mass was 1.05 t 0.09 m2. Surface tension was calcu- lated during inflation and deflation. The results indicate that surface tension approaches zero at low lung volumes or arras and is always lower during deflation than during inflation at a given volume or area. Fi.rhtr. M. l., Wilson, M. F. and Weber, K. C. (Student Fellowship Re- cipienl ) Respiration Physiology 10( 2):159-171, 1970. Otlirr .r'porrr West Virginia }leart Association. From the Department of Health. Education and Welfare, U. S. Public Iicalth Service, Consumer Protection and Environmental Efealth Servicc, Environmental Control Administration. Appalachian Laboratory fo• Occu- pational Respiratory Diseases, and Department of Physiology ard Bio- physics, West Virginia University Medical Center, Morgantown. INFLUENCE OF CIGARETTE SMOKE ON GUINEA PIGS: EFFECT ON PULMONARY CELLS AND SERUM ANTITRYPSIN LEVELS A total o( 50 mak guinea pip was exposed to the smoke from two packs o( cigarettes per da~ for two weeks; this exposure was then reduced to one pack per day for the remaining eight weeks of the exposure period. A correspb++dma group of 50 animals served as control. Measurenxnts were made of the celli recoverable from the lung and of the serum anti- trypsin kvels in both groups of guinea pip. Repeated exposure to ciRa- rette smoke caused a significant increase in the total number of cells recoverable frmm the lung. DiRerential cell counts showed also that animals exposed to ciaarette smoke had an inereased number of po1ymbrphoauclear kukocytes (PMNs) in the fluid obtainod by lava6e of their lunas. This increase was accompanied by no change In serum antitrypsin kvets over the ten-week exposure period. The findins that inhalation of c:6arette smoke results in a sigtifiunt increase in the number o( PMNs recoverable from the lung may be of significance in the pathogenesis of chronic pul- monary emphysema in man. Ffinr, G. [.., Maxwell. K. W. and Renzetti, A. D.. Jr. (Student Fellowship Recipient ) Archivrr of Environmenraf Ntdrh 22(3):366-369, 1971. Otbrr .uProrrr National Institutes of Health. From the Mkrobiok." Rcsearch l.abcxatory, Department r-4 Medicine. Veterans Administntqa ltospital, and University o( Utah College of Medicine, Salt Lake City. 46 THE ULTRASTRUCTURE OF ALVEOLAR MACROPIIAGES FROM 1(UMAN CIGARETTE SMOKERS AND NONSMOKERS Alveolar macropha6es of ciprette smoken and nonsmoken, obtained by endobronchial lavabe, were examined with the ekctron microscope. The macrophages are characterized by surface processes, polymorphic nuclei, an extensive Golgi pparatus, cytoplasmic filanxnts, and various inclusions. These inclusions are membrane-bouoded and contain acid phonphatase. Thus they are probably lysoaonrcs and their derivatives, or6anetles which function in the digestion and storage of pha6ocytosed substances. In Reneral, alveolar macrophages from smokers contain larger and more heterogeneously structured inclusions than alveolar macrophases from nonsmokers. There is, however, considerable variation in the number, size, and heteropenicity of inclusions within individual macnipha6es from smokers, suggesting that various stages of phagicytosis are represented in lava6e samples. Furthermore, sxrx substances which are phaRocytcncd but not digested probably accumulate in the larger heterogeneous inclusions of smokers. Pratt, S. A., Smith, M. EI., Ladnraw, A. l. and Finley. T. N. Laboratory 1 nvtsriaorion 24 ( S):331-33a, 1971, Other aurpsrtr National Inatitutes of Health. From the Departments of Anatomy and Medicine, The University of New Mexico School of Medicine, Albuquerque. THE JUXTA-ALVEOLAR LYMPNATICS IN THE IIUMAN ADULT LUNG: HISTOLOGIC STUDIES IN 15 CASES OF DROWNING A clear definition o( alveolar lymphatica is vital to the understanding of pulmonary clearance of intra-alveolar fluid and particulate matter. In order to study closely the Irmphatics of the adult human IunR, extensive tissue sections were taken (rorn the lungs of 15 persons who died from drowning. These lunga exhibited excellent demarcation and often dilatation of the I7mphatic capillaries. Two main sets of lymphatics were distinguished here: the superficial or pleural set and the deep or p.renchymatoua (peri- bronchovascular) lymphatic piexua. Regarding the disputed occurrrrrce of Iveolar lymphatics, a sharp distinction was made in this study between the air-blood barrier, the interalveolar aeptum an d , d the alveolar wall. No lymphatic capillaries were observed at the kvel of ttie air-blood barrier or the interalveolar septum, but characteristic lymphatic capillaries were present between the alveolar walls nd the Interkrbular, pleural, peri- bronchial, or perivascutar connective sheeta. These were defined as "juxta- alveolar" lymphatic capillaries because o( their cbae topographic and probable functional relationship to the alveolar lumins without being a part of the interalveolar septa themselves. The occurence of these lymphatics might hclp to explain the clearance of intra-alveoiar transudates, eRudates, or inhalcd materials via the lung lymphatics that eventually drain the bhK)d stream. 47

1', Louwtrynt. 1. M, Americon Review of Respiratory Disease 102:877-8R5, 1970. From the Department of Patholo8y. University of Leuven School of Medi- cinc, Leuven, Belgium. ARGYROPHII., FLUORESCENT AND GRANULATED (PEPTIDE AND AMINE PRODUCING?) AFG CELLS IN HUMAN INFANT BRONCHIAL EPITHELIUM. LIGIIT AND ELECTRON MICROSCOPIC STUDIES l.i6ht optical, histochemical and eloclron microscopical studies of 22 newborn human infants have shown that the epithelium of the bronchi con- tains an Impressive number of distinctly ar8yrophil, fluorescent ( aftcr freeze-drying and formaldehyde vapcx treatment) nd ultfastructurally granulated oetls. These AFO cells contain numerous round clcctrlM- opaque dense<orcd vesicks. Intramucoaal nerve endings are slu, present, suggesting a "direct contact" with the AFG cells. It is proposed that these cells in the human lung are related to the growing list of presumably peptide and amine secreting "argyrophil" or APUD. and other cclis which have been reported in different other tissues and organs. A chemoreccptor- like function also may be possible. In addition, it might be that t tie AFG cells could intertcre with various pulmonary areas, such as bronchial smooth muscle tone, pulmonary vasomotion, the circulatory adaptrtion (or failure) of the newborn at birth, and pulmonary oncology (carcinoid and oat cell turnors). Louwrryns, 1. M., Pcuslens, 1. C. and Cokelaere. M. l.ile Scftnces 9, pt 1( 24 ):1417-14 29, 1970. Other arPportt Belgian National Foods voor Wetenschappelijk Onder- zoek. From the Experimental Laboratory of Cardiopulmonary and Genital Pathology. University of Leuven School of Medicrne, Lcuven, Belgium. L'U1.TRASTRt1CTU~ E DE LA,CLOISON INTERALVEOI-AIRE DU POUMON APRE.S CRYODECAPAGE The recently devdoped frccze-etch technique was applied to an elec- tron micro.wQical Investigation of the interalveolar septum and the air- blood barrier of lun6s of adult mice, Suinea pigs, and rats. This bxhnique is different from traditional ekctron microacopy because it avoids some chemical interactions in the t3ssues, exposes tissue strudures in rclicf, and allows the observatioe of membrane surfaces en face. Moat importantly, this technique hero revealod new srorphoio8ic data concerning the lameltar inclusions of the larye alveolar epithelid celh, favoring furthermore the lipoprotcinic nature of tlseae {nc{uaions and their relaticxnhip with the "atveolar anti-atekctatk factor." The arfaoes of Qw lamcllae of these inclusions are studded with small pl.ques of 170 A, which are not ob- served with traditional eleetron m~ro.eopy. The distance between two 48 I 0 I successive lamellae is remarkably constant. Their configuration strikingly resembles the frceze-etch ultrastructure of synthetic lipoprotein miccltes (and of lipid inclusions of yeast cells), suggesting that the small plaques could represent the protein fraction of the lameltar inclusions, while the lamcllae themselves could contain lipids. Lauweryns, 1. and (',ombeer-Desmecht. M. Journal de Microsropie 10( 2):1 J9-148, 1971. From the Dcpartment of Patholo6y. University of Leuven School of Medi- cine, Leuven, Belgium. L't)LTRASTRUClURE DE L'EPITIIELIUM BRONCIIIQUE 1T IfRON('11101.A1RE 1)E LA SOIIRIS 'l hc ultrastructurc of the bronchial and brvnchiolar epithelium and especially of the Clara cells of adult mice has been investigated with the light and the electron microscope (after fixation with osmium tetroxyde). The ultrastructure of the ciliated celh conforms to the classical data. The Clara cells arc characteriud by an extremely abundant smooth endoplaunic reticulum. the absence of cilia and the occurrence of numerous peculiar mitochondria. Some cisternae of the snrooth endoplasmic reticutum flatten out around the mitochondria. The Golgi zoae seems to form some secretory vacuoks. As discussed, these ultrastructural data suggest a secretory function for the Clara cells, but do not allow an identification of the nature of this presumed secretion. Moreover, some aspects of the ultrastructurc of the Clara cells are variable, three different types being distinguishable and dcscribcd, i.e., the "usual type," the "clear type," and the "adult 1ype." It is postulatcd that these types probably represent different functional phases of the presumed secretory cycle, i.e., a young form (the clear type), an adult form (the usual type) and an involutionary phase (the dense type ) • L,ouweryna, 1. M., Cokelaere, M. and Boussauw, L Bulletin de CAssoclallon dtt Anatomittef 146:548-StS0, 1971. From the Department of Patholoay. University of Lcuvcn School of Medi- cine, Leuven, Belgium. THE BLOOD AND LYMPHATIC MICROCIRCULATION OF THE LUNG This cs.oy syathewnes the results obtained while studying one of the most intriguing and perpkxin8 problerro of the human lun6, i.e., its Wood and lymphatic microcircutation. One thing that Is essential and preliminary to an understanding of the microdreulation is some knowledge of the microarchitecture of the lung. This is presented In this paper for both the adult and the prernature lung. Results of correlated macroscopic and micro- scopic studies of the lungs of humans, beagle doR.s, and monsrct do6s are 49

presented. Primarily, this study is concerned with the angioarchitcctural relationships between the bronchial and the pulmonary vascular s)stcmc. The relationships arc distinct in each circulahxy network. The bronchial arteries are directly caxtnected to each rNher, establishing a"parallc'" type of vascular supply. The bronchial veins also form thepc ribrunchial venous plcxus. In the pulmonary circulation, however, the pulmonary arteries and veins form terminal intralobutar branches. The bronchial and pulmonary arteries also communicate in the broachioles by means of a common capillary network. As extensively discuased, the authors were unable to observe bronchopulmonary artenoarterial or arteriovenous anastomrrscs in entirely normal adult IunBs of beagles, but they seem to be normally present in the lung of the human fetus and infant. Probably they disappear during childhood, to reappear easily in the adult lung in the presence o: even minimal pulmonary inflammatory reactions or processes. (x thc lynphatic microeireulation, many problems are presented and care(ully consodcred hut few re answered definitely. Lauweryro, l. M. In Sornmen, S. C. (cd.): Patholoty Annual 1971, New York: Appleton- Century-Crofts, 1971, pp 365-415. Oth.r support: U. S. Public Health Service and the Bclgian National Fonds voor Wetenschappelijk Ondcrzoek. From the Department of Pathotogy, University of Lcuven School of Medi- cine, Leuven, Belgium. AIRBORNE INFLl1ENZA PR8-A VIRUS INFECTIONS IN ACTIVELY IMMUNIZED MICE Circulating antibody in aetively immunized animals can be effective in preventing death, clinical illnen, and residual pathology. In this study of the production of airborne pulmonary virus infections, young adult white mice, immunized subcutaneously with formalized mouse-lung suspensions of influenza PR8-A virus (test mice), along with corresponding unim- munized (control) mice, were exposed to sublethal and highly lethal clouds of freshly harvested homologous virus. The control mice developed rapid and fatal pulmonary infections following exposure to the lethal cloud of virus and extensive andpe rsistin8 pulmonary ehangea following exposure to subkthal viral clouds. 'i~he test mice became only mildly ill and survived the lethal airborne infections, although the virus grew to high titcr in the lungs. Pneumonic involvement was focal and confined to the bronchial air passages and adjacent alveoli. Resolution occurred and residual lung scar- ring was not seen in recovered mice. Thus, while not preventing infection, immunization affected in a higfily favorabk and significant way the clinical course and lung pathdo6ical findings of the mice exposed to airborne in- fluenza virus. Liroifi, C. G., f lertweck, M S. and Hoekwatd, R S. .I rchrves of F.nvirrrnmrnrol llraflh 21(3):333-346, 1970. I Otlh.r support: U. S. Army Medical Research and Development Com- mand, U. S. Public Health Service, Hughes Employees' Give Once Club, and Hastings Foundation Fund. From the Departments of Medicine and Pathology. University of Southern California School of Medicine, Los Angeks, and University of Chicago School of Medicine, Chicago. GLYCOPROTEIN SYNTHESIS BY THE TRACHEA Because glycoproteins make up a mapr portion of the solids of trachcobronchial secretions, their synthesis by slices of rat trache was studied by using radioactively labeled glucosamine as the substrate. Upon incubation the radioactivity was incorporated into protein, that is, material insoluble in cold ctharnd, cold and hot triehloro.cctic acid, and not ex- tracted by lirmd solvents. Incorporation was dose-dependent, occurred at an optimal p11 of 8, and was decrrased in the presence of glucose. A small amount of U-glucosamine-r'C was metabolized to r'CO7. Compared to L-kucine-r,C, the curve of glucosamine incorporation rose less steeply initiall~, and the ascendin~ Iimb assumed a si8rnoid shape. Puromycin rn- hibited incorporation of radioactivity from glucosamine by 44-7296 and from leucine by more than 95%. The microsomal fraction of tracheal homogcnate had the highest specifie activity of the subcellular fractions throughout  four-hour incubation. The akohol-insolubk radioactive product was mostly resistant to hyaluronidase but was made soluble by Pronase. When tracheal slices were incubated with D-Slucosamine-6-aH and were examined by radioautography, the dreatest density of Srairu was found in the epithclium. Yeagcr, if., )r., Massaro, O. and Macro.o, D. American Review of RespLatory Disease 103(2) :188-197, 1971. Other a.pport: American Thoracic Society. From the Pulmonary Division, Veterans Administration - George Wash- ington University Medical Center, Washin8ton, D.C. PULMONARY ARTERY LIGATION: EFFECT ON IN VITRO PROTEIN SYNTHESIS Left pulmonary rtcry ligation was performed in adult mongrel dogs of both sexes. The doEs were killed at various intervah following ligation; lung siices from both the (ipted and non-ligated sides were prepared and incubated and cell-free protein synthesis was studied. Results showed that one week following puu m.nary rtery li6ation in vitro protein synthesis was higher in nonhemorrha8ic tissue from the li6tted side than it was in tissue from the non-li~ ted side. The physiological significance of this in- crease in protein synthevs and the mechanism or stimulus by which it is brought about are not elear. Mauiaro, /).. Wcis, Ff. and White, O. Archives of Intrrnal Medicine 127(S):861-862, 1971. SI 50

Other support: American Thoracic Society. From the Pulmonary Division. Veterans Administration - George W ash- ington University Medical Center, Washington, D.C. DIFFERENTIATING TYPE I AND TYPE 11 ALVEOLAR Cf:L LS IN RAT LUNG BY (hO.-Nal STAINING A histochemical method offering a usable differential stain for the type 11 alveolar epithelial cell is presented here. The fixing-staining mixture used in this investigation consisted of oneQart of 2% aqueous (hO, and three parts o( 3% Nal in distilkd water. Fresh nt lungs were cut into 2 mm slices and immersed in this solution for 24 hours at room tempcra'ure. Controls were fised in buffered OsO, alone. Selective slainin of tyfe II alveolar cells was shown by the (hO,-N.I mialure but was a~+xn1 in Ihe controls No additional staining d the sections was rtyuired, and the sckc- livity was readily observable in either paraffin or Araldite sections by light microscopy and in Araldite sections by electron microscopy. MrNary, W. F.. !r. nd fa-Bermani, A-W. Stain Technology 4 5( 5) 215-219, 1970. Other .upport: Boston University School of Medicine. From Boston University School of Mcdicine, Boston. CHRONIC COR Pl1LMONAI.E: ITS PATHOGENESIS IN APPALACf1IAN BITUMINOUS COAL WORKERS This study presents a quantitalive, postmortem analysis of cardiac and pulmonary vascular structure in 178 Appalachian soft coal workers. A total of 58% of these older bituminous miners with coal worker's pneu- moconiosis had anatomic evidences of moderate or severe chronic cor pulmonale at death. The degree of cor pulmonale correlated best with thepe rcent of lung comprised of abnormal air space which is a measure of emphYsema; the observed emphysema was of both centrilobular and more generalized types. There was no apparent eorrelation between degree of cor pulmonak and the volume o( dust macttks, or the content of bire- fringent silica crystals and collagen In the dust macules. Also, in 10 younger miners medial muscle mass of small pulnxxur7 arteries inside dust macules was 2.3 tinxs thcne outside the macuks. This was due mainly to hyper- trophy of individual muscle fibers. This vascular abnormality helps explain the mild pulmonary arterial hypertension and cor pulmonalc of many younger miners. Nacyr. R. L. and laqueur, W. A. Archives of ParholoRy 90:487-493. 1970. Ott:er.rpportr U. S. Public Ffealth Service. From the f)epartmcnt of Pathology, Pennsylvania State Univcrsity ('ullcge of Medieine, f Iershcy EFFECTS OF SMOKING ON LUNG STRUCTURE OF APPALACIf1AN COAL WORKERS Long-term cigarette smoking alters some structural features of pncu- moconiosis in soft coal workers but has littk effect on the disease process in anthracite miners. lhis appraisal is based on a quantilative, morphologic study of the postmortem ~eart and lung structure of 144 Appalachian unrcrground coal workers. Of these minen, 32 had a lifetime history of nonsmoking. 24 smoked a pipe or cigan, and 88 smoked cigarettes in various amounts. Results s.tiowed that there was no influence of smoking on the primary coal dust maeuk and its direct eompllcations, but the bituminous-coal miners who smoked had a 17% to 33% greater degree of cor Pulmonale, 9% to 25% mor emphyaema, and 56% to 68% mare bronchadar goblct cells as evidence of chronic bronchiolitis than non- smoking bituminr,us-coal miners. No significant effect of snwking on lung and heart struclurc was found In anthracite minen. Notyt, R. L.. Mah<x+, J. K. and Dellinger, W. S. Archives of Environnunral Htalth 22:190-193, 1971. Other .u pport: U. S. Public Health Service. From the Department of Pathology. Pennsylvania State University College of Medicine. !lcrshcy. RANK OF COAL AND COAL WORKERS PNEUMOCONIOSIS This study presents evidence that the much-reported association be- tween coal rank and pneumoconicxis is probably fortuitous. Regional differences in the prevalence and severity of coal workenpne umoconiosis often have been attributed to the rank of coal mined; the higher the rank of the coal, the greater the prevaknce of the pulmonary disease. Here a quantilative, morphologic stud~ was undertaken of lung and heart structure of 312 Appalachian miners who were classified by rank of coal mined. A total of 390 corresponding controls was studied also. Among the miners chronic cor pulmonak was common in all groups. Pulmonary fibrosis in- creased with the number of parenchymal silica crystals and with bronchiolar goblet cell hyperplasia, an evidence of chronic bronchitis. The actual volume of coal dust retained in the lungs, excluding its silica conlent, seemed to have only a small eRect on the development of eor pulmonale and symp- toms of pulnxrnary insufficiency; the prevaknce and severity of pneu- mocomosrs are more likely related to chronic bronchitis and exposure to silica than to rank of coal mined. Noeye, R. L.. Mahon, l. K. and Dellinger. W. S. ,1rnerirnn Review of Rrspirarory Disease 103(3):350-355, 1971. Other tupportr U. S. Public llealth Service. From the Department of Pathokrgy, Pennsylvania State l)nivcrfily ('ollege of Mcdicine. Hershcy. 52 53

ADRENAL GLAND STRUCTURE AND THE DEVELOPMENT OI: IIYALINE ME:MBRANE DISEASE There is a demonstrable relationship between adrenal gland structurc and the development of hyaline membrane disease in newborn infants. Analysis of 3R7 consccutive autopsies on human neonates showed that adrenal glands were 19% lighter in infants with hyaline membrane disease than in those without the disorder owing to a greater number of adrenal cortical cells in the latter infants. A positive correlation was found between the presence of infection arising before birth and the absence of hyaline membrane d'neasc, the infected infants havin4 larger adrcnal glands. Since surfactunt appears to play an imPortant role in hyaline membrane disease, it is of interest that anencephalic neonates who had little or no adrenal fetal cortical zones and halt-siud adult zona had 45% the mass (A osmiophilic granules inpu Imonary type 11 alveolar cells as did non-ancnco phalic control infants. llre oaniophilic granules are reporledly the anatomic representation of surfactant. Naeye, R. L., Eiarcke, It. T., Ir., and Blane, W. A. Pediarrict 47 ( 4) 650-657, 1971. Ot/1rr support: U. S. Public Nealth Service and Health Research Council of the City of New York. From the Department of Pathok.gy, Pennsylvania State University College of Medicine, 1-fenhey, and Department of Pathology. Columbia University College of Physicians and Surgeons. New York City. ACQUIRED ATTENUATION OF CHEMORECEPTOR FUNCTION IN CHRONICALLY IIYPOXIC MAN AT IiIGI-1 ALTITUDE Chronic exposure to h) po xia during adulthood produces alterations in a person's control of breathing. Measurements of the resting ventilatory response to hypoxia in eight persom native to high altitude. 10 long-term noo-natjve high altitude residents, and 10 low altitude controls show th.t high altitude living results in a diminsbad ventilatory response to hypoxia. The degree of attenuation is related to the length of time spent at high hitude. Similarly, hypercapnk ventilatory drive is reduced in non-native high altitude residents as It Is b a peater degree in high altitude natives. In contrast with these findings at nat, induction of hypoxia during exercise produces in both groups o( highlanden an increase in ventilation com- parabk to that in the controls. This susgesls that there is a marked dis- crepancy between hypoxk response during exercise and hypoxic drive at rest, which is presumably mediated by peripheral chemoreceptors. The observation here that hypoxic ventilatory drive is almoat completely absent while hypcrcapnic drive is only partiall~ reduced parallels closely the gcnerally accepted view that the pctipheral cErcetoroceptors are resdortsibte tcx virtually .11 of the resting hypnxk drive nd roughly half of the hyper- capnic drive in man. This su~s Is that the alterations In ventilatory control at high altitudes are due to tailure of peripheral chemoreceptor function. 54 Wril, I. V., Byrne-Ouinn, E., Sodal, 1. E., Filley, G. F. and Grover, R. F. The Journal o/ Clinical Investigation 50( I):186-195, 1971. Ot6.r .rpport: U. S. Army, Department of Nealth. Education, and Wel- fare, U. S. Public Itcalth Service, and American Thoracic Society. From the department of Mcdicine. University of Colorado Medical ('cnter, Denver. V. Tissue and Organ Culture DIFFERENTIAI. CYTOLOGICAL AND CYTOCNEMICAL RESPONSES OF VARIOUS CULTURES FROM MOUSE TISSUES TO RE:PEATEI) EXPOSl1RF-S TO PUFFS FROM THE GAS PIIASIi 0F CIIARC-OAI. PILTERED FRESH CIGARE:TfE SMOKE A total of three types of primary cultures, namely kidney tissue, em onic lung organ, and lung explants from Snell mice, and one estab- lis cell line, 3T3 cells from Swiss mice, were used to assay the biological effects o' the gas phase from charcoal-filtered fresh eibarette smoke. Re- pe.ated exposure to puffs of the g.n phase of charooal-filtered fresh cigarette snioke did not evoke a significant alteration of growth in primary kidney tissue and embryonic lung organ cultures from Snell's mice. There was also no effect on Snell primary lung explanb, and on their outgrowing macro- phages and epithelioid cells. There was, however, an effect on 3T3 cells, in that exposed 3T3 cultures showed an enhancement of gowth, of mitotic index, and of cellular atypism, including variations in DNA content. In none of the exposed cultures was a morphological cell transformation ob- served. Lruchfenberatr. C. and Leuchtenberger, R. Experimental Cell Research 62(1):161-172, 1970. Oth.r.rpport: Association Sui..e des Fabricants de Cigarettes. From the Department of Cytochemistry, Swiss Institute for Experimental Cancer Research. Lausanne. TISSUE CULTURES In this first of a series of monographs on athetoaclerosis, the field of tissue culture is examined from its inception through its differentiations to its future possibilities. There are seven sections of this book as follows: 1. Background; 11. Vascular organ cultures; 111. Non-vascular tissue cul- tures; IV. Vascular tissue cultures; V. Cell aggregation; VI. In vivo vas- cular tissue cultures; and VII. Review. Section IV, the longest division of the book, considers such aspects of this discipline as history, methex1oloRy, quantitative aspects of cell cultures, mixed cell populations, single cell pop- ulations, lipoid transport by cells, metabolism, eytophannacology, cytotuxi- cology, cylogenetics, and atherogenesis and in vitro cultures. SununaririnR original findings and surveying the work reported in the literature, this 55 I 65100 6855

informative, succinct book incorporates all the features of an easily read tissuc culture textbook. PnJlek, O. 1. TIJSur CufrurfJ: MonoRrnphs on Athtroxttrosir, Basel: S. Karger, and Baltimore: Williams & Wilkins Co., 1969, vol. 1, pp 1-143. OrA.r .rpport: 1)elaware Fleart Asaociation, Dover Medical Research ('enter, Lilly Research Laboratories, and U. S. Public Hcalth Service. From thc Dover Medical Research C.eater, Inc., Dover, Det. V1. Stttdlt•s at the CeUcetnrLeorl STREPTOVARICINS INEEIBIT RNA DEPENDENT DNA POLYMERASE PRESENT IN AN ONCOGENIC RNA VIRUS The strcp tovarkin ~pkx is an extremely potent inhibitor of the reaction by which DNA ia transcribcd from the RNA template reside nt in purified murine leukemia virions. Strrptovaricin C and rifamycin SV are also quite active, while stttpldydipn and ritamp~cin are relatively poor inhibitors. In experiments reportod here, the M.Eoney strain of mnrinc leukemia virus was grown in JLS-V9 cells and purified using a duublc sucrose gradient centrifugation. Electron microscopy of the purified lrac- tions revealed 203 X 1011 p.rticka/ml.; the protein concentration was 0.5-0.75 m6/ml. The results o( inhibition studies showed that the strr.pto- varicin compkx, a mixture of seven macrolides (plus undetermined ;om- ponents), is a potent inhibitor of the MLV polymerase reaction. More than 75% inhibition Is obtained with eoncentratioru as low as 40 r.p/ml. Complete inhibition is not obtaired, however, even with concYntrations of 400 ra/mI. Slreptovaricin C, a pure macrolide, was also quite active but less so (on a weight basis) than the complex. Rifamycin SV gave an inhibi- tion similar in magnitude to that of /treptovaricin C. Broekman, W. W., Carter, W. A., U, Li-El., Reusser, F. and Nichol, F. R. Norure 230( 3291) :249-250, 1971. OtA.r arpport: U. S. Public Nealth Service and American Cancer Srxiety. From the Departments of Medicine and Microbiology. Tfie Johns I1opkins University School of Medieine, Baltimore. COMPARATIVE PRODUCTION OF INTERFERON BY NUMAN FE:T'AL, NEONATAL, AND MATERNAL CELLS This report compares interferon (IF) production in nine primary human fetal cell lines with their rng maternal cell lines. Ran- domly selected neonatal tisaue, obtai txtween the fint and third days rr/ extrauterine life. wat studied alao. IF production was measured after exprnure to two inducers, virus (Ne..cast disease virus) and synthetic, double-stranded ribopolynuckotide (poly r1:tC). Results suggest that fetal cells display, in tissue culture, increased responsiveness to both inducers. Fetal cells released 300-fold more IF than maternal cells on exposure to poly rl:rC. This enhanced capacity for IF production was consistent in cultures developed from fetal skin obtained between the 10th and 20th gestational wcck. The response was relatively stable, persisting in cells cultured for 18 generations (about 14 weeks). On infection with New- castle disease virus, fetal cells produced, on the averase, 4 to 6.5 times more IF than maternal or neonatal cells. The virus was adsorbed with equal efficiency br each type of cell. Increased production is apparently independent of the rates of overall protcin synthesis, since fetal and maternal cells have very similar rates of total protein synthesis. Carrer, W. A., Irarxk, K. R., Essien, B., Prochownik, E. and Kaback, M. M. Infection and Immunity 3(S):671-677, 1971. Other .wpportr U. S. Public Health Service, American Cancer Socicty, Maryland Division, and National Institutes of Health. From the Departments of Medicine. Microbiology and Pediatrics. The Johns Hopkins University School of Medicine, Baltimore. STRUCTURAL REOUIRPMENTS OF RIBOPOLYMERS FOR INDUCTION OF HUMAN INTERFERON: EVIDENCE FOR INTERFERON SUBUNITS The binding of aN labeled poly I:C-DEAE-dextran complexes to cells has been measured. Low temperatures were used to separate the binding reaction from later steps involved in cell uptake. About 0.5% of uncomplexed poly IC is bound at 2' after one hour; 20 times more of the macromokcular aggregate is bound when DEAE-deatran is present. Thus, this enhancer potentiates human interferon (IF) release by delivering more of the active poly IC complex to the triggerin site. The IF proteins induced by both virus and synthetic ribopolymen tave been purified and their apparent subunit structure detected. As an initial step in the characteriza- licx+ of IF's, the IF'a have been extensively puriBed by means of the new technique of electrofocusina in polyacrylamide gels. In gel chromatography of unpurified IF's, mouse IF behaved as  single molecular species with a MW of 38,0(X). Human IF cluted predominantly in a position correspond- in6 to a MW of 24.000. Isoelectric focusing of mouse and human IF was performed on the IFs obtained from gel chromatography. Data from thesc studies strongly suggest that IF exists as a dimer of two similar, perhaps identical, subunits. The monomeric unit of human IF has a MW of 12,000. The different isoeleetrie points for the dimer and monomer suggest that certain surface charges must rearrange during oligonier fornra- tion, but without influencing activrty, since there does not seem to be  measurable cooperative effect between the subunits in the dimer frxm. f he mechanism of thc conversion is currently under study. 56 57

Carter, R'. A. and Pitha, P. M. In Been, R. F.. Ir., and Braun, W. (cds ): Biol:,gical F.B«-rs of f's,ly- nuclmridrs, New York: Springcr-Vcrlag, 1971, pp R9-105. Othrr.rpport: U. S. Public Nealth Service and American Cancer Society. Maryland IKvision. From the Departments of Medicine and Microbiology. The lohns Hopkins l)nivcrsity School of Mcdicine, Baltimore. MEMBRANE-ASSOCIATED DNA IN THE CYTOPLASM OF DIPLOII) HUMAN LYMPNOCYTES To determine whether nonmitochcxdrial, cytoplasmic DNA exists in human lymphocytet and to investitNe its subcchular hocalizalion, cellular DNA was uni(ormt~ labeled with (2-"C)thymidinc, and cells wrrc lyscd with the rxsnionic dctergent NP-40. This detergent lyses the plasma mem- brane, but not thc nuclear membrane, as shown by phase-contrast micro- scopy; nuclei can be quantitatively removed /rom lysates by low -spced centrifugation. The lysate obtained after removal of nuclei was separated into several fracti<>.n. Virtually all cytoplasmic DNA was found in a frac- tion sedimcnting at 13.000 X` and of this less than 10% was surercoikd. This cytoplasmic DNA.  sptcies that is apparently associated with the plasma membrane of diploid human lymphocytes that are continu(ously growing, is described. This DNA differs from nuclear or mitochc.ndrial DNA by virtue of its location in the cell, time of synthesis in the cell cycle, nd various physical propcrties. At present, the role of this DNA is un- known. Lerner, R. A., Meinkt, W. and Goldstein, D. A. Prarrcdints of the National Acodrmy of Sciences 69 (6):1212-1216, 1971. Other .rp'ortr National Institutes of Health, National Cancer Institute nd American Cancer Society. From the Departments of Experimental Pathology and Microbiology, Sciipps Clinic and Research Foundation, La lolla, Cal. OXYGEN TOXICTfY IN TItE NEWBORN GUINEA PIG LUNG: TIiE INCORPORATION OF TRITIATED URIDINE INTO MONORI BOSOMES WeprvQose that oxygen interferrs in the ecology of cellular ropula- tions of t eh lung and have designed a variety of approaches to this thesis. In this experiment, newborn guinea pigs were poisoned in 100% oxygen for four dars. When monorihosomes prepared from their lung homofenates were examined, using a pulsed T•uridtne label in vivu and sucrose giadicnt ultracentrifugation technique, the results sug~csted that rilxru+me synthesis per surviving cell was probably increased. It ia the fourth day alur which niarks the dcvchipnxnt of bronchiolar mucocal necrosis, thc bcpinninR of I bronchiolar metaplasia, the appearance of anomalous ribosnmal protein, and a severe depression in DNA synthesis. Thus, the emergence of a new cell population is tied to certain molecular changes apparently induced by a toxic environment, and is followed by restructuring of the lung in a permanently distorted pattcrn, which includes emphysema. The observations raise fundamental questions about the nature of regeneration in the lung ksbuk, particularly Ihe role of cell populations, and suggest a need for improved experimental models based on molecular techniques. An incidental and surprisinE discovery was the sharp absolute reduction in ribosomc synthesis o( controls and experiments by the fourth day of life. Bicber, M. M., ('ogan, M. G., Durbridge, T. C., and Rosan, R. C. BiuloRia Neonatorum 17( I-2):35-43, 1971. Oth.r ..rpportr ('alifornia Tuberculosis and Nealth Association. From the Ikpartmcnt of Pathok.gy, Stanford University Medical ('enter, Palo Alto, Cal. VII. Plnarmacu[osy and Ptlychoplwrrnacotogy ANTIASTIIMATIC ACTION OF SYMPATHOMIMETICS: A REVIEW OF TFIE LITERATURE ON TNEIR BRONCHOPULMONARY EFFECTS In this review paper, sympathomimetic bronchodilators are grouped into specific categorics based upon the extent of their activity on adrencraic receptors. In the bronchopulmonary system, there are two types of adrener- gic receptors: the alpha receptors and the beta receptors. These receptors can be activated either directly or indirectly by the release of catechola- mines. The end result of stimulation, either direct or indirect, of both alpha and beta receptors is a reduction in airway resistance, which is the basis of the antiasthmatic effect. On the grounds of their stimulatory actions. the sympathomimetic brnnchodilaton can be divided into four classes: direct stimulants of alpha and beta tcceptors, Indirect stimulants of alpha and beta recepton, direct stimulants affecting predominantly olpha recepton, and direct stimulants affecting predominantly beta rcccpton. Among the 16 sympathomimetic bronchodilators which are useful in the treatment of bronchial asthma are: (I) Direct alpha-beta stimulant: epinephrine; (2) Indirect alpha-beta stimulants: ephedrine, phenylpropanolaminc; (3) Direct alpha stimulant: phcnykphrine; (4) Direct beta stimulants: II drugs in- cluding isoproterenol, dioxethedrine, mctaproterenol, soterenol. and sal- butamol. A vJado, D. M. Journal of (7inic al Phannacoloay and the Journal of New l)rugi 10(4): 217-221,1970. From the Ikpartmrnt of Pharmarnlugy, University of Pennsylvania Sch(Krl of Mcdicine. Philadelphia. 59 5R

0 HORMONES AND PULMONARY EFFECTS OF TOBACCO: CORTICOSTEROIDS IN ANESTHETIZED DOGS Pulmonary compliance and resistance were measured in 36 anes- thetized dogs in this study of the interaction between corticosteroids arxf cigarette smoke. A single in~ection of either hydroccxtisone or dexametha- some caused a sli6ht but dcfinite fall in pulmonary resistance to low-r levels. These corticosteroids probably exert their influence by local relax+- tion of smooth muscles of the airways. Relaxation of the do6 s bronchial smooth muscles by corticoateroida could not be attributed to potentiation of the sympathetic bronchodilator mechanism or to inhibition of bronch.r constrictor mechanisms. Changes In the mechanical properties of the lung were eticitcd in response to isoproterenol, compound 4R-R0 (a histamine liberator), serotonin, histamirse, Inhalation of 5% oxygen in nitrogen for Iwo minules, and inhalation of eiprelte smoke. Inhalation of SiRaret)c smoke, alorx, increased pulmonary resistance In anesthetized dogs. i lrc respemac of the dogs to cortietxteroida tiven before exposurc to ciRarct:e smoke was so variaMe that it was Impouible to determine the relationsh~p between cigarette smoke and airway sensitivity after corticcnicroid therapy. Carrilto, L. R. and Aviodo, D. M. Archivrr of F-nvir,nrvruntot f/colrh 21(2 ):149-153, 1970. From the Department of Pharmacolo6y. University of Pennsylvania School of Medicine, Philadelphia. CARDIOPULMONARY EFFECTS OF PROGf:STATIONAI, AGENTS IN EMPIIYSF.MATOUS RATS Prosesterone and mcdroxyproRrsteronc (in a dose 1/50 that of progesterone) prevent the development of experimental emphysema in the rat. They do not, however, increase the rate of survival from the initial experimental procedures of tracheal liption and intratrachcal injection of phytohemagglutinin. The studies reported here show that in the rat with ligation of the trachea, which received intratracheal injections of phyto- hemaoulinin but no progestational aeent, the following effects were mr served after six weeks: (a) high functhmal residual capacity; (b) high percentage of air sQaces in histological sections of the lung; (c) cardiac output; (d) normal pulmonary arterial pressure and pulmonary vascular resistance; (e) extreme ekvalion of pulmonary arterial pressure in rc- sponse to rapid infusion of saline; and (f) normal electrocardiographic pattern and normal Ihietness of veetricular waW examined at postmortem. The concurrent administration of ptvr sterone or medroxyproResterone prevented (a) and (b) but did not affect the other factors. Rats with tracheal occlusion alone had abnormally high levels of pulmonary rtcrisl T ressure associated with hypvala and relieved by inhalation of high oxygen. hesc rals had not progressed to the stage of chronic cor pulrrxrnale, although those under (e) above indicatcd scxne change in distensibilily of the pulmonary blood vessels. 60 Inoh, T. and Avrodu, A Af. Clrcsl 59 ( 6 ) :659-666, 1971. From the Ekpartmcnt of Pharmacolo6Y. University of Pennsylvania School of Medicine, Philadclphia. INCREASED RETE?NTION OF NOREPINEPNRINE-aH IN VAS DEFI:RENS DURING NERVE STIMULATION This study represents an attempt to determine quantitatively the im- pr~rtance of increased uptake during nerve stimulation of the isolated hypo6aslrie nerve vas defercns preparation by studZins the action of drugs known to block the resting uptake of catecholamines. Retention was in- crcascd when isolated vas deferens of guinea pigs were exposed to norepi- ncphrine-'If during nerve stimulation of the hypo6sstric nerve at a rate of SO shrKks/sec; 1.5 mscc pulse duration; 4 see/min for 60 minutes. 'This increase in rctention of norcpinephrine-'ll remained unaltered when the program of stimulatiom was prolonged to 10 sec/min or decreased to 2 sec/min. It was inhibited when cocaine, desmethylimipramine, or phenoxy- bcnzamine was added to the orsan bath 10 minutes after norepinephrine-'E1. These results indicate that uptake is increased in order to meet the need for replacement of catecholamine lost during high-impulse activity. This increase was not obscrvcd when monoamine oxidase inhibitor was present in the organ bath. BhoRc1, B. and Zcidman, N. American Iourncl of Physiology 219(3):691-696, 1970. From the Department of Physiology, St. Louis University School of Medi- cine, St. Louis. INFLUENCE OF CIfRONIC ADMINISTRATION OF NICOTINE ON TFIE TURNOVER AND METABOLISM OF NORADRENALINE IN THE RAT BRAIN Chronic administration of nicotine accelerates the rate of disappear- ancc of intraventricularly administered 'H-noradrenaline from the rat brain. '1Ttis is associated with normal levels of 'H-normetanephrine sug- gesting an increase in intraneuronal deamination. The slope of noradrena- line decline in nocotine pretreated rats is sigtifkantly greater than that of control animals, while steady state levels of brain noradrenaline are about equal in both groups of rats. Tlth sugges ts an inereased turnover rate and utilization of noradrenaline induced by nicotine treatment. Also, the actions of sedative, antidepressant, and central stimulant drugs in the ral brain are not affected b~ nicotine pretreatment. Amphetamirx, reserpine, and acetylcholirx sig++rficantly decrease the endogenous catechotamines in the rat brain, but dccrease in .mine level is the same in nicotine treated and control animals. The results of these experiments indicate that chronic administration of nicoline increasn noradrenatine turnover in the brain and possibly may increase Ihe deaminalion of this amine. 61

Bhagor, B. Psychopharmacoloyia 18(4):325-332, 1970. Orh.r srpport: National institute of Mental Ifealth. From the Department of PhysioloFy. St Louis University School of Mcdi- cine, St. I.cwn. EFFECTS OF CfIRONIC ADMINISTRATION OF NICOTINF ON DRUGINDUCEh IfYPNOSIS IN MICE The present study was undertaken to determine the effect of rhronic administration of nicotine on the ph+rmaoological responsivily of mice to selected central nervous system depressants. Barbital, which is excreted mainly unaltered, was chosen as an indicator for alteration of CNS sensi- tivity on the rate of drug entry into the brain. Ifcr<obarbital, pentub.uhital, zoxazolaminc and ethanol were used to test the alteraticm in the drug metabolizing enzymes. Chronic administration of niastine in micc for six weeks did not affect the duration of hypnosis induced by pentobarbital, hexobarbital, barbital or z.oxaxofamine. The onset of barbital hypnosis remained unaffected. These results suggest that chronic administra ron of nicMine did ncx affect the drug metabolizing activity of hepatic microsomes. While psychotropic drugs significantly potentiated the hypnosis due to Pcntobarbital and hexobarbital, the degree of potentiation was the same in both chronically nicotine-prttrealed mice and controls. Nicotine (+re- treatment significantly increased the duration of sleeping time caused by ethanol alone Howevcr, when treatment of nicotine continued for a longer period of time, the duration of hypnosis induced by ethanol returned to control level. Bhaaar, 8., Bayer, T. and Lind, C. PsychophormacolnRia (Brrt.) 21( 3 ) :287-293, 1971. Ot14rr anprort: U. S. Public Health Service. From the I)epartment of Physiology. St. Louis University School of Medi- cine, St. Louis. SOME NEUROCNEMICAL CORRELATES OF ALTERED MEMORY CONSOLIDATION In this roundup of memory consolidation experiments, several ncurcr- chemical changes attcnding dtered retrograde mrxsia in response to electroconvulsive shock (ECS) art explored and related to modifications o( the memory consolidation procesa. An initial series of experiments was concerned with the extent to which changes in brain serotonin levcl, as approximating those produced by ECS. would affect the incidence of cnn- ditioncd response acquisition. Other experiments tested the role of I) the serotonin precur.or, S-hydroxytryptophan; 2) al;e diRcrencc%; 3) difTercn- tial howsinR; 4) rrduccd brain amine kveli; and 5) aminc chanRrs al the ccllular level thc studies summarized here tauggC1t an imlwntant role for I S-hydroxytryptaminc in the mediation of at least one phase of thc mcmory amsolidation prrKCss. The relationships between changes in this amine in whulc brain, in regional sites within the brain, and on a cellular Ievcl, nd certain other cvcntc implicated in otherphases of the memory consolidation pnxcss (RNA synthcsis, protein synthesis, and their inter~.)cpendency) have been shown, under several experimental conditions. These rclalicxr- ships may constitute a modcl within the framework of which serotonin may play an important role in normal memory consolidation and in those conditions wherein the memory consolidation process in impaired. E.«mon, W. B. Transacrions of the New York Academy of Sciences, Serirs ll, 3 2( 8): 948-973, 1970. OrArr support: National Institutes of Health. Frum the Ikpartmcnt of PsychobRy, Oueens C<>Ilcgc, City Elnivcrsily of Ncw York, I Iushing. TIIE ABILITY OF VARIOUS NICOTINE AGENTS TO REt.E:ASE ACETYLCHOLINE FROM SYNAPTIC VESICLES Nicotinic agents apparently penetrate the intraccllular site of thc nerve ending causing the release of endofenous acetykholine (ACh); non- nicotine agcnts do not. Evidence to thrs effect is presented in this study of the release of cndogcnous ACh from synaptic vesrcles, the bixly's primary storehouse of cndogcnous ACh. In these experimcnts, mixtures of rat cerebral cortex vcsiclcs and nicotinic and non-nicotinic drugs were incu- bated at S" C for five minutes to study the release of ACh by these drugs. Nicotinic agents such as nicotine, tetramethylammonium, cholinc, rd carbachol released ACh effiiciently. Non-nicotinic agents such as tricthyl- choline bromide (TEC), hemicholinium bromide (IiC-3), and DMAE among others failed to show the sarne effect. The ACh-relcasing effect of nicotinic agents was not blocked by TEC. HC-3, or DMAE. 'fhcsc results suggest that the nicotine agents are taken to an intracellular site of the nerve tissues in order to cause rekase of endogenous ACh from synaptic vesicks. The antiniccHinic agents bfock the uptake of niaNinic agents at the membrane site but not at the intracellular site of the nerve tissucs. This is demonstrated by the fact that TEC has been shown to block the nicotinic effect at the ncuromuscular junction and autonomic funglion in intact nerve terminats, but not the rtkase of ACh from the synaptic vesicles localized within the nerve tissues. Chiou. C. Y., l.rrnR, 1. P., Potrepka, R. and Spratt, 1. L. Archivrs lntrrnariunoltt de Pharmocod ynamit tt dt I hfrapir 187 ( 1): 88-96, 1970. Otlh.r support: 11. S. Public Health Service. From the 1)rpartmcnt of Pharmacology. University of Iowa ('ullcFc of Mcdicinc, Iowa City. 63 62

.1 ~ a~ tn .- m m STUDIES OF DMAE-INDUCED POTENTIATION OF TIIE PRFSSOR AND ('ONTRACTILE RESPONSES 10 EX(X;L.NOEIS NORE?PINEPIIRINE IN NORMAL AND RESERPINE-TREATFI) ANIMALS While it has been reporled that DMAE, a hemicholinium derivarive, enhances the pressor and contractile responses to exogenously administered and endogcrK,usly released norcp nephnne, the mechanism nf thcx actions has not been clearly defined. This paper presents evidence that DMAE:- induced potentiation of the responses to electrical stimulation of adrentrgic nerves and direct-acting amines is mediated by an increased release of neurotransmittcr from adrrnergic netvt endings. EJsing control and rtser- pinc-trcated cats, blockade of the adrenergic ncuronal uptake nxcha.ism on DMAE-induccd pMentialion of the conaractik and presxx respt nses of the cat nictitating membrane to eto~t ncwsly administcrcd and cnJcgen- ously released ncxcpinephrine was evaiuated. Cocaine pretreatment autag- onired the ability of DMAE to enhanct the responses to exogenous r..tc- pinephrinc. DMAF-induccd potentiation of the responses to ncrvc stimula- tion was unaltered after cocaine administration. These results suggest that DMAFE releases rxxepinephrine taken up by the adrenergic nerve terminal after the administration c.f norcpincphrinc. Grccnbcrg. S. and l.ong. l. P. Arc-hivrt Inrrrnarinnolrr de PharmocoQvnomic et de Thfrulric 141 ( 2): 365-375, 1971. From the Department <.f Pharmacology. l)niversity of Iowa Collcge (rf Medicine. Iowa City. EFFECTS OF NICOTINE ON SELF-STIMULATION IN RAT'S Nicotine facilitates self-stimulation in rats. In experiments reportcd here, rats were trained to press a lever to receive intracranial electric stimulation through a stereotasically implanted bipolar electrode in two different areas of the hypothalamus. Nicotine was given i.p. at several dosc levels. The effects of nicotine were found to be rate-dependcnt. Facilitatitm of responding for self-stimutation was observed at 37.5 rg/kg and higher doses of nicotine in rats with low response rates. In rats with high rctponse rates, nicotine had very little effect or even caused depression. Nicotine had no appreciable effect or unreinforced responses and it counteracted the depres%ant effect of barbituratcs on sdf-slimulation. The facilitatory effect of nicotine on self-stimulation resembled that of amphetamine in many respecls. Facilitation by nicotine was reduced or blocked by mecamylamine, a tertiary Fanglion blockinq agent, and could ncN be produced in anintals treated with rescrpine 24 EKwrs earlkr. It Is propcncd that nicotine, by acting on a central nicotinictExrlinergic rrceplor, may indirectly cause release of norepincphrine which. in turn, produces the facilitatory effcct on sclf-stimulati<m. 64 Prodhan, S. N. and E3owling, C. Thr lournal of Pharmacofogy and Experbntntal Therapeutics 176( 1):229- 243, 1971. ' Other .u pport: U. S. Public Ifealth Service. From the Department of Pharmacology, Howard University College of Medicine, Washington, D.C. EFFECT OF COLD STORAGE ON TEIE RESPONSE OF GUINEA-PIG TAENIA COLI TO CERTAIN CATECNOLAMINFS AND OTHE:R A(;ENTS Cold storage affects smnoth muscle responses to sympathomimctic amines and other active agents in several complex ways. In the prescnt csperimcnls, rclaxatirm proluccd by phcnykQhrrne, epinephrirx, nrxcpine- phrinc, and isc.prtHcrcnc>t on guinea-pig taema coli was not modified after cold storage of up to six days. The responses of the cold stored strip to the first three of these drugs were prevented by pretreatment with alpha-bkxk- ing agents but not by treatment with beta-blocking agents; the opposite was obs,rved for the response loesopr otercnol. On the other hand, the con- traction produced by nxthacholine, physostigmine, KCI, nd Ba was dccrcased after three to six days cold storage. Similarly, the relaxation produced by ATP and 3',S'-cyclic AMP was also decreased after cold storage. Incubation in low Ca medium had no effect on the inhibitory action of catecholamine used in cold stored strips but suppressed the re- sponse to ATP and 3',S'-AMP. It can be seen that in this experiment the cold stored strip still responded well to catecholamincs but not to ATP and 3',S'-AMP; moreover, alpha- and beta-adrcncrgic receptor blocking agents did not modify the response to e><ogerrous ATP and 3',S'-AMP while such blocking agents prevented catecholamine response. These data suggest that the relaxation mechanisms of catecholamincs may be different from that of exogenous high energy substances. Shibara, S., Hattori, K. and Timmerman, D. European lournalo/ Pharnracwlogy 11:321-331,1970. Oth.r support: U. S. Public Health Service and ltawaii Eicart Associa- tion. From the Department of Pharmacology, University of Hawaii, Nonolulu. ADRENERGIC INNERVATION AND COCAINE-INDUCED POTENTIATION OF ADRENERGIC RESPONSES OF AORTIC STRIPS FROM YOUNG AND OLD RABBITS The use of the rabbit aortie strip as a pharmacotogic tool has become very popular in studying adrenergic mechanisms. In this histological study, a calecholaminc-spcci6c fluorescence was observed in the tunica nxdi of the aortae of ytwng rabbits but not of old rabbits. After five days nf codd storage at 2' C or reserpine treatmcnt, these specific fluorescent sut*- 65

stances disappeared from all layers of the tissue; such treatment decreased the tissue norepinephrine (NE) content in both young and old rabbit aortae. Checking the NE content of the tissues by a spedrophotolluoro- metric method, the aortae of young rabbits were found to contain twice as much NE as those of old rabbits. Cold storage blocked the contractile response to nicotine and tyramine. The sensitivity of the young rabbit aortae to nicotine and tyramine was significantly greater than that of old rabbit aortae. Despite these diRerences. NE-induced contractions were potentiated by cocaine rn strips of both types. Cocaine-induced potcntiation could still be demonsirated after 5 days of cold storage. No cocaine po- tentiation of NE-induced contraction eould be seen after incubation for 20 minutes in a Cs {4- -lree medium and treatment with MN r+ anwj Co+ +. Cocaine potentiated the Ca+ 4 -induced contraction in a Car r-free mcdium with a high potassium content. These result suggest Ihat adrenerRic nerve fibers penetrate into the smooth muscle layer of the rabbit uxtac, but that the caxaine-indueedpotentiatiexr may not be dependent on these drenerRic nerve terminals in the aortic media. External Ca +4 may possibly play a significant role in the development of cocaine-induced potenliation. Shihara. S., Hattori, K., Sakurai. 1., Mori, l. and Fujiwara, M. Journal of Pharnronrloty and Experimental Therapeutics 177( 3):621-632, 1971. Other .rPrort: U. S. Public flealth Service, Elawaii Heart Association, and American Medical Asaociation Education and Research Foundation. From the Department of Pharmacology and Pathology, Univc rsity of Hawaii Sehooi of Medicine, Honolulu. TEEE EFFECT OF COCAINE ON CATECHOL-0- METHYj.TRANSFERASE AND ON THE RESPONSE TO NOREPINEPIIRINE OF RABBIT AORTIC STRIPS Although the oooeentrations of IUa M and 10• M cocaine caused the potentiation on the norepinephrine-induced contraction of rabt+t aortic st~pa. such concentrations did not inhibit ntechol-0-methyltrsnsferase (COMT) from rat liver. The concentrations of 10 • M and 10 6 M pyro- gallol which potentiated the norepuxphrine-induced contraction si6nifi- cantly decreased COMT activity. Cocaine, but not pyrosallol, po.entiated the phenylephrine-indueed contraction of fresh strips and norepinephrine- induced contraction of 4 days cold stored strips. The data indicate no positive evidence to sn the theory that eocaine-induced potentiation is mediated by the inhi 'tory action of cocaine on the COMT enzyme system. Hattori, K. and Shlbatt, S. lapantsr /ovrnal o( PArr+.avlosy 21:SS9-362, 1971. Ot14.r.rPprtr U. S. Public HeaEtlt Sertlce and Hawau Ifcart Association. From the Ekp.rtnxnt of Phhrm.ooEopr, University of lfawaii Sch<«d of Mcdicine, Hooolulu. THE RELATIONSNIP BETWEEN EXTERNAL CALCIUM CONCEMfRATION AND THE RECOVERY RATE OF AORTIC STRIPS FROM NICOTINE TACHYPHYLAXIS The depletion of endo6enous eateeholamine stores by agents which act through catecholamine-rekase mechanisms is considered to be the mechan- ism responsible for the development of lachyphylaxis. (;nfortunalely, the nature of nicotine-induced tachyphylaxis is not clear. In rabbit aortic strips a repeated exposure to the same eoncentralion of nicotine produced tachyphylaxis. Complete recovery occurred SO-60 min following the wash- out of the first application of nicotine. Incubation in medium containing norcpinephrinc ( 10 9 M) or dopamine (10 t M) did not modify the time course of recovery. . High Ca }+-medium halved the recovery time and low Ca4 +-medium inhibited this process. Thus nicotine tachyphylaxis does not appear to be due to catecholamine depletion but rather to be a blockade of the adrener6ic nerve effeetor site by depolariz.tion which prevents rK)re- pinephrine release. Shibata, S., Elattori, K. and Sanders, B. European Journal o/ Pharn.aeoloay 16:109-112, 1971. From the Department of PharmacoloEy. University of Hawaii School of Medicine, Honolulu. CNS EFFECTS OF THE ALDEHYDE PRODUCTS OF BRAIN MONOAMINES Theprrsent series of experiments show that S-hydroxyindoleacetalde- hyde (S-EITA) and indokacetaldehyde (tryptaidehyde) are as quick actin9 and as potent as aerotonin and tryptamine in inducing electrophysioloaical and behavioral effects in experimentd animals. Other related compounds are also pharmacologically active but higher doses are rrquired to demon- strate their effects. In these studies, indokacetaldehydes and indokethyla mines induced sleep in newly hatched chicks, tryptophol depressed mouse exploratory behavior, and high doses of phenylacetaldehyde induced CNS depression in mice and chicks. As reEards rabbit photic evoked potentials. S-EITA first enhanced and later depressed the slow negativity and en- hanced the fasl positive compkx (similar effect to serotonin). Hiah doses of S-hydroxytryptophol. 5-hydroxyindoleacetic acid, and serotonin de- the slow negativity. In contrast to hydrox~mdoks, tryptamine and indoleacetaldehyde augmtn ted slo.r rseptivity. The monoamine oxidase inhibitor, par6yline, reduced the eReeh of setotonin and 3-HTA on photic resportses, and phenylaoetaldehyde depression of rnouse exploration. Further experimentation will be required to confirm or rule out a significant role for the cndoRerrously formed aldehyde derivatives of brain amines in behavior and in the action of monoamine oxidaae inhibitors. Sabclli, 1E. C. .nd lliardina, W. l. (Grantee: !. E. P. Tornun) Biological Piychiarry 2:119-139, 1970. m 67 66 a> m ~..

Other aupport: l1. S. Public Health Service. From the Fkpartmenl of Pharmacology, Chicago Medical School. Chicago. THE INFL(1ENCE OF INHIBITORS OF AMINE METABOLISM ON TFFE EFFEC IS OF SEROTONIN AND ITS METABOLIIES ON PIIOTIC EVOKED POTENTIAL IN RABBITS Non-arxtthetired rabbits were used to test the effect of intraventricular injection of scrotonin (SHT), 5-hydtoxytryptophol (5HTOL), and 5- hydrosyindoleacetic acid (SHIAA) on the photic evoked potential. Fast potentials were depressed by the three 5-hydroxyindolcs tested at all dose levels; this depression was more pronounced with SIiTOE- than with SFiIAA or SHT. 51tT reduced the slow negative wave (SNW) amplitude at all doses tested. Tbe onset of the SNW depression occurred 15-20 minutes after injccticm of 0.03 mg of SIIT, and it was preccded ,1y a transient enhancement of the SNW. The pharmacobgical activity o( 511TOL and SFIIAA, together with the delay in the onset of the SNW depression induced by low doses of SHT, suggests a possible participation of 5117 metabolites in SHTs central action. The dose response curves for the three hydro.yindoles showed that 511TOL and 51iIAA were as potent as 511T. In addition, pretreatment with two different inhibitors of aminc metabolism, N-bcnzyl-N-nxthylprop-2-ynylaminc (pargylinc) and tetra- ethylthiuram disulfxk (1 FTD), markedly altered the cftects of 5111' with- out inffucncing the effects of its metabolites. Sabelli, H. C. and Giardina. W. 1. (Grantee: I. E. P. Tvman) Eiptrirntia 27 ( 1 ) :64-65, 1971. Othrr.rpprt: U. S. Public Ffealth Service. From the Department of Pharmacology. Chicago Medical School, Chicago. CHRONICALLY ADMINISTERED NICOTINE AND TItE BLCOD PRESSURE OF NORMOTENSIVE AND RENAL IFYPERTENSIVE RATS Chronic administration of nkotirx in the drinking water of rats, in amounts roughly correspond mg to the e9uivaknt of one or two packs of cigarettes dail~, ekvated the mean systolic pressure. When this trcatment was prolonged, however, the pressor effect was reversed and a depr-ssor effect was ultimatel~ obtained. With large doses (three or four packs of cigarettes) only a depressrx effect wa observed. Withdrawal of nic.itine during the depressor phase produced a marked, fluctuating, and prolonged pressor response. In renal hypertensive rats, administration o( cither thc "low" or the "high" doses o( nicotine Fowcred the systolic pressurc below control levels. This effect was not observed until the neurogcnically main- tained phase of renal hypertensbn was cstablished. An observation of potenual rclevancc to the rok of .moking in human hypcrtensi,m was the marked pressor rcb„und and the prolonged period of recovery altcr with- drawal of the highcst dose of nicotine. With such a pressor responsc, it is likely that the heavy smoker who attempts to break his habit may undergo a prolonged pcricxF of elevated pressure. Wrnzrl, !). G. and Azmeh, N. Archivrs Inrrrnariunalrs de Pharmocodynamir rr de Tlrrrul,ir I 97( 2): 367-376, 1970. From the Department of Pharmacology and Toxicology. The lJnivcrsity of Kansas School of Pharmacy, Lawrencc. A STIIF)ENT EXERCISE IN CLINICAL PHARMACOL(X;Y: RENAL EFFECTS OF DIURETICS A group of 122 medical students tested the renal effects of either a placebo, hydroehlor(ithiazide In four doscs, acetaiolamidc, triamterenc, or the combination of hydr<xhlonNhiazkk and triamterene. The student cxcr- cise was designed so that the following information would t.c provided: a) a comparative effectiveness of the various diuretics, b) a dose-response relationship, c) information concerning different sitcs of action, and d) the efficacy of a drug combination. Results showed that there were no apparent diffcrences with respect to the incidence of side effects between the placebo and the various medications. Also, there was no apparent relationship be- tween the response of the smoker vs. nonsmoker within any drug group. Each of the medications was associated with an increase in urine volume. In general there was a dose response relationship with hydrochlorothiazidc with the 6, 12, and 25 mg doscs. Howevcr, the 50 mg dose of hydrochloro- thiazidc did not produce an increase in urine flow greater than the 25 mg dose. No advantage of the combination of hydrochlorothiazide and tri- amterenc was found. Gilmore, 1. P., O'Brien, W. M., Brand, E. D., Peach, M. 1. and Wesr/all, T. C. Current Therapruric Research 12( I 1):759-769, 1970 Other support: U. S. Public Health Service. From the DeQartmcnts of Pharmacology and Preventive Medicine, Univer- sity of Virginia School of Medicine. Charlottesvilk. i V111. Metabolic Studies THE KININ-FORMING SYSTEM IN PLASMA Since there is existing evidence of vascular permeability occurring ind,pendrntly of known mcdiation >:ystems, it has become esscntial to search for other ryns,ble mechanisms that induce vascular injury. Amung the humoral mechanisms that must he considered in the mediation of such injury is the kinin forming syitenr. In this study, a rwmtxr (if agents were uscd to activate dic kinin system. 68 69

m ~ m B Three components of the kinin forming system have been purified in precursor form. l he first, Hageman Factor, is a molecule of 90,(XX) daltons which upon activation cleaves into subunits of 30,0(1() which activate the second comporxnt, prckallikrein. The active llagcman Factnr acqtires enzymatic activity. Prekallikrein is activated by limited protedysic, being ckaved into three and two subunits, respcctively, for human and rabbit prekallikrcin. Upon activation, kallikrein cleaves the vasoactive nona- peplide, bradykinin, from purified kinirqgen. Activation of llagcmrn Factor was achieved on vascular basement membranes by enzymes cleaving arginine esterase activity and by negatively charged particies. Cochrarsr, C. G. and W uepper, K. D. Insnwnopotholoty o/ /n/larnn.etlort, Exaryta Medics International Con- `ress Series No. 229, pp 137-145. April 1971. Oth.r ..Pprrr U. S. Public Health Service, National Muttipk Sckrnsis Society and American lleart Aa,ociation. From the Department o( Experimental Pathology, Scripps ('linic and Re- search Fovndation, le )olla, Cal. METABOLIC FACTORS AFFECTING PROTEIN SYNTIIESIS BY LUNG IN VITRO The addition of glucose to lung slices incubated with Ieucinc-"C increases the protein specific activity of the acid-insoiuble material present in the reaction mixture. Glucose causes no increased degradation of acid- insoluble radioactivity nor dexs it cause an increase in the tissue acid- soluble radio.ctivepcol when kucine-"C or a-aminoisobutyric-I-"C acid are the substrates. Ilus effect of glucose on kucine-"C incorporation into protein is relatively greater under anaerobic conditions although total in- corporation is markedly decreased. Galactose, fructosc, and ribose do not augment amino trcid Incorporation into protein as handily as does glucose. Neither palmitate nor carnitine added to the reaction mixture causes an Increase in the amino acid incorporation into protein, even though camitine doa'signifieantly augment the oxidation of palmitic acid-"C to "CO_. The effect of malonic acid on protein s nthesis with and without glucose is of intereat as it may reffoct on the mechanism by which glucose augments ptotein synthesis. Makxtk acid markedly depresses formation of "CO_ when gJucoae-6-"C is the substrate but causes only a mcxiest depression of ' 4COi ptoductirxt when 6luonse-l-' IC is the substrate. This and several related observations suggest that the glucose effect is mcdiatal through the pentose phosphate pathway rather than the tricarboxylic acid cycle. Massaro, D., Simcxr, M. R. and Steinkamp, 14. lournol of Applied Physloloty 30(l ):1-6, 197 1. Oth.r .rrr.rt: American Thoracic Society. Frnm the Pulmnrury I)ivisinn, Veterans Adnrinistration -(:corgc N'a.h- ington University Medical Center, Washington. D.C. 70 I I I STRESS AND CIGARETTE SMOKE EFFECTS ON LUNG MITOC1fONDR1AL PI-IOSPHORYLATION Male guinea pigs were exposed to cigarette smoke on a 2'h day acute exposure regimen. To insure mouth breathins, the nares of these animals were occluded with warm molten paraffin or with  quick-setting liquid silicone rubber. Sham-exposcd animals were subjected to the same condi- tions of nose plugging, confinement, and machine noise without cigarette smoke. The stress of forced mouth breathing and the superimposition of cigarette smoke on this stress caused losses in oxidative phosphoryl.tion etTiciency in the guinea pig lung mitochondria In vivo. Nowever, bss of efficiency was greater in the mouth-breathina, air-inhalin6 animals than in cigarette smokers under the same conditions. The "tishtness" of r hosphorylation coupling at mitochondrial sitp 2(cytochrome rcductase vcl) and 3(cytrxhrome osidaae level) was measured In these experi- ments, using artifxial electron generators or acceptors. The ntouth-brcath- ing nonsmoking animals lost eff'iciency at both sites 2 and 3. Mouth- breathing smokers lost efficienc~ onlr at phosplwrylation transfer site 3. These stress and cigarette smoke eRects at sites 2 and 3, respectively, suggest that the losses of oaidative phosphorylation efficiency are due to different mechanisms acting at specific loc- i within the mitochondrion. Kyk, 1. L. and Riesen, W. f!. Archives of Environmenro! llealth 21( 4):492-497, 1970. From the Life Sciences Research Division, IIT Research Institute, Chicago. IX. CheniLtry and Biochemiu" GEL PERMEATION ANALYSIS OF MACROMOLECULAR ASSOCIATION BY AN EQUILIBRIUM METHOD Macromokcular association by gel permeation can be determined by an equilibrium method. In this paper, a thermodynamic description of the simplest case, dimerization, is presented in terms of the equilibrium con- stant for association nd equilibrium partition coeflicients for specia present. The theoretical analrsia yields an equation relating the equivaknts of monomer in the external phase to total tquivalents, from which the ectuilibrium constant for asaociation ma~ be obtained. Those parameters, critical in experimental design. gel swelling time, and centrifugal collapse point are determined for Sephadex gels of porosity G-2S to G-200. Appli- cation of the method to human hemoafobin gives a value of Kh - S X I(1' for dissociation of oxyhemoglobin at pls 7.0 in 0.1 M buffer. CanKron, B. F., Sklar, 1.., Grcenficld, V. and Ad1cr, A. D. Separation Scirnce 6( 2): 217-22d, 1971. Other.rpport: Connecticut Ncart Association. 71 m DD m (.J

From the New England Institute, Ridgefield, Conn.; Department of Internal Medicinc. Univcnity of Miami School of Medicine and the Lahuratories for 1lcrnalological Rescarch, Iloward Hughcs Mcdical Institutc, Miami, Fla. BIPHASIC BL(x7D OXYGEN DISSOCIATION CURVES IN 1(EMOGLOBIN S FIEMOGLOBINOPATHIES. SICKLE CELL HETEROZYGOTE:S The purpose of this study was to evaluate blood oxygenation in hcmo- globin S heterozygotes (A-S and S-C) and, by comparison with hlo%xl-hank bkiods, to evaluate the effect of 2,3-diphosphoglycerate (2,3-1)P(;) dcplc- tion on oxygenation. In order to do this, the oxygen hindina cr.rves of whole cells rd hctcrozygrws sickle cell hcmog)obinr>frathies wcr! nwasured. Undcr the conditions of mcawrement (T=25', p11=6.4, pCO,--0) the curves are biphasic. Samples In which 2,3-DPG is very low or ab.icnl are nxm(phasic, suggesting that there is a diffcrential binding of 2,3-f.)P(i to S and non-S hemoglobin in cells of hcterozygotes. The biphasic curvc has been confirmed in a hemoglobin SC patient, under physiologic conditioms (T-37', pEt-7 4, pCOY-_ 44 mm). Camrron, B. F. Bio<hrnrical and Binphysi(a! Research Communicorions 43(4):888-892, 1971. From the Dcpartment of Internal Medicine. University of Miami Schrn)l of Medicine and thc Liboratorics for Flematologic Rcsearch, Howard FluFhcs Medical Institute, Miami. FIa. PHOTO-OXIDATION OF BOVINE OXYMYOGLOBIN IN FROZEN SOLUTIONS. THE EFFECT OF REDOX ACTIVE INORGANIC ELEMENTS IN MUSCLE EXTRACTS ' The problem of oxidation of myoglobin in aqueous muscle extracts is cornplex. This work, done on bovine mtack extracts, emphasizes the effect of redox active substances in muscle on the photocatalyud autoxida- tion reaction, primarily at acid pH where the conversion is from the red MbO. to the brown MMb. In these studks, photocatalyzcd autoxidation of MbOr ~ MMb in frotcn extracts was found to obey first order kinetics as found for Mbn1 0( sperm whale and horse. Formation of MMb-OII in crystallized beef MMb was studied. Its pK of ionization 8.90 is closel~ similar to that of sperm whale and horse and diflcrs from that rs( camcl. Among the unbound elements examined in beef muscle extracts, iron was mrnt responsible for photo-oxidation of MbO_. EDTA (_ iron cauud further increase in ox0ation; EDTA akxse, however, caused protcction In a related experiment, a metmyogk.bin cationic reducing agent wac isolated from dcprorcinircd horse and bovine muscle extracts by ion exchange chromatography Assaf, S. A., Bratzler, 1., Cameron, B. F. and Yunis, A. A. Comparative Biochemistry and Physiology 39B( 2): 395-407, 1971. Other support: U. S. Public Heallh Servrce and The Michigan Agriculture Experiment Station, East Lansing. From the Departments of Medicine arid Biochemistry. University of Miami School of Medicine, and the Laboratories for Hematological Research, E'oward Hughes Medical Institute, Miami, Fla. INTERFERON: EVIDENCE FOR SUBUNIT STRUCTURE Mouse interferon was purified over 500-fold by differential precipita- tion, centrifugation, gel chromatography, and a new technique of elcctro- Irxusing in ~yacrylamide gets; human interferon, subjected to the same cxpcnmental procedures, was purified about 15(X)-fok1. Ekrth interfcrons, although initially homogenous on gel chromalography, displayed two molecular forms (A and B) on ekctrophoreais in pll gradients. In mouse interferon, Form B(isoekctricpa nt (p1) 7.35) had a molecular weight of about 38,000 and Form A( p1 7.15 ) which was equally active, a molecular weight of 19,000. Purified Form B was dissociable into Form A, but the reverse reaction occurred to a much ksscr extent. In human interferon, Form B(pt 5.60) had a molecular weight of about 24,0(IV and Form A ( p1 5.35) which may contain up to 85% of the total activity, a molecular weight of 12,000. Both forms appear to be equall~ active. The dissocation of both human and mouse (nter(erorss into subunits appears to take place during dialysis vs. low salt. The data are consistent with the idea that the native molecule exists as  dimer of similar or identical subunits. Dimer Eormation, which probably occurs within the cells, does not seem to lead to a measurable cooperative effect between the subunits. Carter. W. A. Proceedings of the Natlonal Academy oJ Sciences 67 ( 2):620-628, 1970. Other support: U. S. Public Health Service and American Cancer Society, Maryland Division. From the Departments of Medicine and Microbiology, The Johns Hopkins University School of Medicine, Baltimore. PURIFICATION OF MOUSE AND HUMAN INTERFERONS: DETECTION OF SUBUNIT STRUCTURE Mouse and human interferons, virnl-induced, initially behaved as single molecular species in the studies reported here. Mouse intcrfcron was purified over 500-fold by differential precipitation, centrifugation, gel chromatography, and isoekctric focusing; human Interferon was purifkd about 15(X)-told. At one step during purification (Istxkctric focusing), two forms of each protein were detected. These two forms were characterized then as to molecular weight. Form A(mouse) has a molecular weight of 19,000 and Form A (human) a molecular weight oE 12.(XX). 1'urm B 73 72

(mouse) ha.s a mokcular wei~fi t of 38.000 and Form B(human)  mokcular weight of 24,000. 'T')rese data strongly suggest that interferon exists as a dinxr of two similar, perhaps identical, subunits. Thc convenion of Form B into A is promoted by the exposure to low salt. In the presence of 0.05 M NaCt, little evidence for two molecular forms exists. Cartrr, W. A. Preparative Biochrmirrry 1( 1):SS-75, 1971. t?rt;.r .upp rr: U. S. Public Health Service and American Cancer Srxiety, Maryland Division. From the Departments of Medicine and Microbiology, The lohns Ifupkins University School of Medicine, Baltimore. SYSTEMATIC STUDIES ON TFTfi BREAKDOWN OF p,d-I)17T IN TOBACCV SMOKES Although DDT was rtmovcd fe 1968 frvm the list of recommended compounds for use on lobacco, these studies on the pyrolysis of DDT are rrkvant' today beause they throw light on the mechanisms of the reactions involved in the degradation of Qesticides in tobacco smokes. This papcr is divided into two aections. Section I deals with the pyrolysis of p,d.Dl)T in a nitrogen atmosphere at 900' C and subsequent isolation and identifica- tion of its degradation products. Mechanisms for the formation of these compounds are proposed and on that basis prediction is made on the formation of DDT degradation products when DfYT-treated tobacco is smoked. Section 2 deals with pyrolytic degradation of DDT iocorporaled in tobacco. Methyl chloride, bis-(pchlorophcnyl) methane, pjS-dichlorobenzo- phenone, p,0-DDM, trans-p."ichk+rontilbene, p.1S-DDE, p,d-TDE and p.d-DDT were idcntifkd as the pyrolysis products. Based on the type and nature of pyrofysis obt.ined, it was corxluded, amcx+g other things, that hydrogenation is one of the most important reactions taking place :n the smoking of tobacco, presumabl~ in the burning zone itself where large amounts of hydrogen are availabk. CJropra, N. M., Dornanski, ). J. and Osborne, N. B. Br("te zur TabakJorschrort S( 4):167-174, 1970. From the Department of Clxmistry, North Carolina Agricultural and Tech- eical State Univenity, Grtensboro. •Tln ~wr...'. 9a0w1. d a. .~.+. .... nr.~a .. .u .r, .w. u. ..e.. ....1ooe. a• r ef OOr. CMr."adr. rY p.MCY. Y.Mll Iw ..r. SYSTEMATIC STUDIES ON THE BREAKDOWN OF p.ff-DDT IN TOBACCO SMOKES: 11. ISOLATION AND IDENTIFICATION OF DEGRADATION PRODUCTS FROM THE PYROLYSIS OF p,{S-Df7T IN A NITROGEN ATMOSPHERE This first systematic investigation into the breakdown uf 17.0 -1)1)T in cigarette main-stream and sidostream smokes was started in the fall of 1967. Two resultant papers already have reported the results and mechan- isms of the breakdown of p.d-DDT in a nitrogen atmosphere, nd in p,p-DDT treated tobacco smokes; this paper presents the methods em- ployed in the pyrolysis of d-DDT and the isolation and identification of its pyrolysis products. p,dDDT was pyrolyzed at 900' C in a nitrogen atmosphere and the pyrolysis Qroducts were collected in pentane at -80' C and isolated by fractional distillation and chromatography on alumina and Florisil columns. The products isolated were: p.d-DDT, p,d-DDE, p.d-TDE, bis (p-chforophcnyl)chkxomethane, bis-(pi.idoroptxnyl)meth- ane, p.d-dichlorobiphenyl, a,p-0ichbrotoluene, heaachloroelhane, chloro- benzcne, tetrachloroethykne, trichloroethykne, carbon tetrachloride, chlor- oform, and dichloromethane. The solid (first eight) pyrolyais products were identified by gas chromatography and infrarrd tromctry, and the liquid (the last sia ) pyrolysis products were identi ed by gas chroma- tography nd colkximetric tests. Chopra, N. A4. and Otborne, N. B. A nalyricaf Chrmisrry 43 ( 7): 849-853, 1971. From the Department of Chemistry. North Carolina Agricultural and Tech- ni •al State University, Greensboro. I EXCRETION OF NICKEL COMPOUNDS BY THE RAT: BLOOD AND URINE LEVELS Nickel and many compounds have been shown to be carcinogenic in animals. Since the route of administration does not seem to be important, this study was inaugurated to see whether nickel solubilizes in tissues and, if it does, what constitutes the route and rate of excretion from the experimental animal. For this investigation, male Fischer-344 rats weighing approximately 260 g were divided into groups of three each. The treated rats were injected intramuscularly in both hind kgs with 30 mg of nickel- ocenc, nickel acetate or nickel powder, all suspended in 0.2 ml of tri- octanoin. Control animals were injected with an equal volume of the vehicle only. All three - nickel powder, the acetate, and nickelocene - were rapidly solubilized. Within 24 hours nickel ions were detected in moderate concentration in the urine and also in the blood during this period of time. The serum proteins were separated ckctrophoretically and nickel was found to be bound to the albumin fraction. For detection and analysis of nickel in biological material, a coforimclric mcahod was em- ployed using quinoxaline-2,3-dithio) as the reagent. Chen, 1. K. M., Tfaro, R. T. and FuiJr, A. The Wasmann lournal of Bioloay 29( I): I-1 S, 1971. OtAer.uppert: 11. S. Public flcalth Service, Carric-Baum-BrowninR'frust Fund and the de Martini Estate. From the Institute of Chemical BicAoRy, University of San Franci.cu. San hrancisco. ° 74 75 a a. u~

X. Epidemiology PREDICTING I.ONGEVITY: METIIODOLOGY AND CRITIOUE This book has two main purposes: first, to advance the state ri( knowl- edge of the methodology of longevity research and second, to increase the understanding of multivariale data analysis methods for social research in general. Aging is a process that interests all of us, but the technology and methodology necessary to adequately study this subjcct have been woefully lacking for years. In this book about longevity, chapters are devoted to Review of thc Literature; Generating the Data; Univariate Procedures; Linear Discriminant and Regression Proeedures; Factor Analysis; and Nonlinear Procedures. The research reported here was undertaken in con- neetion with the Normative Aging Study of the Veterans Administration in order to irkntrfy prcdictcxs of aging and kx+gevity which could bc built into the prenpective design of Ihe study. The results showed substantial change in aignificance of independent variables as one went from univarrale to multivariate analysis, and from linear to nonlinear analysis. The substan- tive findings suggest that physiul predictors are more important than social predictors to account for advanced kmgevity, while the contrary is true lor moderate kmgevity. A major probkm is the control of the secular effcct, and a secular-free, yet "one shot" design, is formulated. Rose, C. L.. and Brff, B. Predicting Longevity: MrrhodoloRr and Crinqur, Luington, Mass.: L). C. Heath and Company. 1971, pp 1-265. (?tlirr sapp rt: National Institute of (hild Llcalth and Human facvclup- nxnt tsd Normalive Aging Study. Boston. From the Veterans Administration Outpatient Clinic, Boston. COMPUTER BASED DATA ANALYSIS IN LONGEVITY RESEARCH Tha paper is primarily concertxd with the application of multiple procedures for data nalysis to a given data set in order to masimizc the surch for meaning." The data studied consisted of the life hishsry in- fcxmation gathered from next of kin listed on death certificates of 500 white male deaths in Boston during 1965. A total of 70 variables was analyzed by zero-onkr correlation, multiple regression, linear diuriminant analysis, factor analysis, nonlinear discriminant analysis, and nonlinear clustering. The results show a substantial shift in significance of independent variables as one procceds from univariate to muhivariate and from linear to nonlinear analysis. Various perturbations in such studies are outlined, such as the effects of secularity and bimodal distributions. Although social variables are more vulnerable to secularity than physical variables, thcrc was some evidcnce of their being more important than physical v;rri:rthlcs in accounting for mexfente longevity. Enslcin, K. and Rrnc, C. L. (Grantee: B. Befl ) ('omputns and lirumcdkaf Research 3(4):289-329, 1970. Oth.r anpport: National Institute of Child Health and Iluman Dcvelop- ment. From the Genesce Computer Cenler, Inc., Rochester, N.Y., and Normative Aging Study, Veterans Adminislration Outpatient Clinic, &ntun. PET ASSOCIATION WITH SELECTED HUMAN CANCERS: A FIOUSE1fO1-D QUESTIONNAIRE SURVEY Two questionnaire studies, one done in early and one in late 1969, surveyed the dcgrcc of famil exprnurc to domestic cats, dogs, and para- kccts in 530 selected "index' eancer households and 530 control house- holds. Bccausc lympfKxna, kukentia, and sarcrxna re{+rescnt the tylies of cancer in animals known to be caused by the C-type RNA tunKU vrruscs, "index" cancer cases were seketed from these diagnostic categories. In the early survey, failure to match sets for duration of residence led to a bias which accounted for an apparent excessive exposure to cats in families with lymphoma. This sampling bias was corrected in the second survey. Overall, no difference in exposure to eats, do6s, or parakeets was found between the index patient households and matched control hcwscholds. No statistically significant difference existed in the number of families with these pets, the average number of pets owned over the past decade, the average number of years that pets had been owned, the degree of handling of pets, or types of illness in household pets. No differences were found in the following condiliorts of cats: abscesscs, fever, fur balb, lumps, bum , malignancy, worms, and anemia. Also, no differences in similar co itions were found in do¢s. Of the family members with cancer, 70-RO% did not own cats. It is therefore unlikely that cats nre a causal `actor in the generality of human carxers. Ilanes, B., Gardner, M. B., Loosli, C. G., Ifeidbreder, G., Kogan, B., Marylandcr, Ift and liuebner R. J. lournal of the National Cancer I nstitute 45 ( 6):1 155-1 162, 1970. Other support: National Cancer Institute. From the fXpartment of Pathology, University of Southern California School of Medicine, L,os Angeks. CURRENT INFORMATION ON FELINE AND CANINE CANCERS AND RELATIONSHIP OF LACK OF RF.LATIONSFIIP TO HUMAN CANCER It is most unlikely that cancer in cats causes cancer in man. While we know that the domestic cat has a high natural prevalence of lymphom and of dctrclahlc ('-type virus expression in the fornr 44 antigen and ('-tyPc patticlcs, cpidcmirdugic nd limited immunologic studies to datc fad to 76 77

suppnrt spread of thcse feline C-typc tunxx viruscs to dog or man. Whcther these viruses spread even from cat to cat under natural conditions is not establishcd. It appears that, under natural conditions, feline C-type viruses are inherited as indigenous latent genes which may activate in the fctus rir later in life. The susceptibility of certain human cells to in vitro infcction with these viruses does not imply infection in viva However, this possibility cannot be denied until immunologic studies are done with serok.Eic reagents of greater sensitivity and sperificity. The domestic cat may prove useful for study of the effects of clxmicals, including those in the environment. The newly found RNA-dcpendent DNA polrmerase may aid investigations in vitro of products of the feline C-type viral genome activation and its governing molecular mechanisms. cardnrr. M. e. lournd of the Narkrnal Cancer I nitfrurr 16( 2): 2g 1-29(), 1971. Othrr.rpporf: Natiortal Cancer Institute. From the fkpartment of Patho{ogy. University of Southern California School of Medicine. Los Angeles. RELATION OF POVERTY AND RACE TO ANTENATAL INFECTION More children of poor (amilics die during the few weeks surrounding birth than do children of the ncx+-pcor. While undcrnutrition is a demon- strated component of this exceuive perinatal mortality, the present study shows that antenatal infection is a factor also. Analysis of 1,(MI consecu- tive autopsies on stillborn and ncwborn in(anta in New York City showed that 27% of these babies had congenital pneumonia or sepsis (or both), primarily related to the aspiration of infected amniotic fluid. Infants from the poorest families had the hi~at percentage of chorioamnionitis, ptrcu- monia, and sptenilis. With escjt improvement in economic status the rate of inflammatron decreased. Rates of inflammation were highest in blacks and io:vcst in whitq; Puerto Ricans had rates just above those observed in whites. Infants with anatomic and cultural evidence of infection had a mean gestational age o/ 29 .vecks, and those without infedion an agr of 32 weeks. The infected infanta were not retarded in growth, but they dk1 exhibit adrenal-gfand alxtortnalitia: adrenal glands were 19% heavter in the infected infants, owing b itsereased cytoplaamk mass of individual cells in the permanent none d the aland. Noryt, R. L. and Blanc, W. A. The New Enjfand lovrnof o/ Mrdk•tr.e 283( t 1) :SSS-560, 1970. Other .rpporfr U. S. Public Health Service. From the fkparlment of Patlatoa, Pennsylvania State University ('ollege of Medicine, Iiershey, and the Dcpartrnent of Pathokigy. Columbia llni- venity Colkge of Physkians and Surgeons, New York City. i Active Projects Following is a list of the principal investigators or institutions of projects under way or activated during the year ended June 30. 1971, together with the respective project tiltes. Completed projects are listed in a later sectinn. t'RINCIPAL INVF_9TICATOR OR INSTtTlfftoN DOMINGO M. AVIADO. M.D., rro- Jriwr of lharmarnloty. University of Pennsyl..nu School of Medicine, Phil- .delphu. PRO)EIT TITLE ERect+ of nkdinc and cisarene .rrwkins on ncuroacnic mecA.ni.ms in the luns Toierance of lunp to tob.cco amoke with .pecial reference to pulmonary empbr. .ema and r..cul.r occlusive k.k.n+ Bro.cbv..cular eflecte of cis.ueue unoke S7EPHEN M. AYRPS M D., Dirrcror• Measurcrned of alveolf,r-arttrial nituoscn Cardropulmonary rebor.tary, St. Vin- difference by gas chromatopaphy cent'. Hospiral, New York City. OSCAR /. BALCHUM, Pw D. Hurinp lroJruor o/ Mrdirinr. University of Southern California School of Medi- cine, Loe Anaeter. FREDERIK B. BANO, M.D. rro/rasor ant CAairman. Drpartrntnr of rarAo- Siolog'• The )ohnm Hopl rnt Univetdty School of Hygiene and Public Hcaltb, Baltimore. BENJAMIN BELL, M.D. Dlrrrror of the Normative Aging Study. VA OW- p.tknt Clinic. Bo.ton. (Initiated rnder C. L. Row.) SAMUEL BELL ET. M.D., Dirreror. DI- risiow of Cardiology. PhU.dclptri. Oew- tr.l Hospit.l, Pbii.delptua. The diffusing capacity of the alveolar membrane in pulmonary emphysema Relation of air pollution to development of chronic pulmonary disease TAe eRects of measured small amounts of carbo.yhe"lobin on cardiweapiray tory function in man ERect of tobacco nnokina and community air lution on human myocardial mcta inn Dettrminantf of the course of emphy- .ema• tNonchit if Interaction of viruses with muco.s of the reapirNory tract, and an andysis of the eRects of tnvironmeM.l variations on Ise function of the mucocrli.ry .yacm Social predictors of longevity T1a eR.ct of nicotine on cardiac Irrita- tion in the pexnce of rc.erpine. and tle e/feA of nicotine on coronary blood tbo, in dosp with coronary insuAkitncy EReds of aiedla on the murphnloay of ooevw.ry arterks and aort.: librrn>tytic sRectft of niootine on human and ani- wd dawn.; enectr of cessation of .mokjna on .erum che.kuerol k.ctr of elronie rrnoker.; the e11ec1s of cla.reue .noke on /rce lelty acid krrls of wb ieets with n+yocardr.l inf.rctionr TDe effect of nirorine on vumw2 p.r.m- etees of cardiovascular function 79 78

PRIN(:1 f•At. INV F:STI(:AT(yR OR tWSTITUTII/N R(1DHD1 V PHA(7AT, Pw [l . Prolrjun of Phrrwudncf, St I oun lin.versily SchMd of Medicine. St 1 o.ris. RICIIARD 1. RIN(i, M D. Prr.Jruor of Mrdrerwr, l)ni.ersity of Snulhern Ca1i- fornia School of Medicine. I us Angeks; Rrrrrrh Auoci.tr, ('alifornia lnvirWe of Technoloty; ()irrr ror uf C'rdrolory .nd Inrr.mrrd Medicine. Fluntington Memorial Hospital. Pasadena, Cal. DIO-RESFARCFI CONSl1T.lANTS. INC.. Cambridge. Mau. RIO-RESFARCN INSTTTUTF_ INC_ Cambridge. Mass RAReARA R. BROWN, PIr.D.. Chir/, t rl.rrrnvnr.i PsirMorrr, Veterans Ad- minrslralron lloaptal. Sepulveda. Cal. SRl1CF F CAMF.RON, M D_ PH D_ Hor.rd Hrlrhrs fnvurrr, Uni.ersir~ o/ Mrami School of Medicine. Miami. WII I IAM M VIN (-ARTFR, M D., Arr/u.nr Pr.rf1sror ul M.dK/nr .nd Mwuhr.d"ty. ll.r lohrn Hoplins lJni. versrry School uf Medicrne. ~•Itimpe. PROJECT TITI.F. The methanism of nicolrnc inalucrJ rr kasc of catecl.olammea FRccl of cigarette %molinR and nicrdine on thc disposition of catecholamrnra in esperinxnul coronary rtery rnsufhcrenc) The effect of smoking on the coronary blood Bow and certain phases of myu cardial mtubolism in patrentb with arterio.ckrWic or hypcrtenarve cardirr- vasculv di.caae Studies in cellular physiology of hrart muscle MeawantM of coronarr bl«.J flow by meanr of r.dio.ctr.e .I(wmrn lAe effecl of smoking on crxonary birx+d flow in pNients with artericsckrwrc heart disease and the enect of nicotine on sloragc of amincs in heart musck Mcasuremenl of eaonary blood flow with  sywem using coinciJence eewntmR; the eRect of nicotine nd change in heart r.le on cardiac mclahdism and related subjeets Studies on cardiac metaholivn with spe cul refrrrnce to myocarJut anu+u TTK influence of mcolrne on lipid compoutron of vascular wall uf the coronary vesaels; its eflect on Ji.trrhW.on of c.,r onary llow Fvatualion of the go1.kn Syrian hamster as a model animal for tobacco smoke inhalation studies Comparison of direct mouse skin espc- sure to whok, fresh cigarette smule and mouse skin painting with smoke condcrnaletl Nicoline and smoking elTecrs on elcctro. encepha"ram eurrelates of behavior. emotionat responsiveness and visual pcrceptnrn in cal. Eaplor.tion of Ihe JiRerences in FFG pattcrn - aubicclivc slate awrclatcs in /mo\er and nonsmoker suhjccts Spectroplo(ometric assay of carlwm mon osrde anJ nitric oxrde hemoglobin (1'ne ~' ny and Ihe anmiviral acliun ,+f in Ie.f crarn 80 I rRINCIrAL InVESTICATOR OR INSTITUTION JACK CHALON, M D, Auurur Pro(ra- aoc of Anrarhraiology, Albert Eirurcin College of Medicine. New York City. SANFORD CHODOSH, h/ D.- Aasiu.nl Pro/raaor of Medicine. Tufts University School of Medicine. doslon. (Initialed under Maurioe Segal, M.D ) NAITER M. CIIOPRA. 1'+r D_ Pro/ra- aor of CArnrbrry, North Carolina Alpri- cuMwal and lecAnical State l)ni.ersdy, (ireensboro. CHARLES O. COCHRANE. M.D., ScriPPs Cf/n/c and R.rraircA Forwd.- /ion. La Jolla. Cal. ALLEN S. COHEN. M.D_ Aaa1»ant Professor o/ Mrdiciwr, Uni.ettNy oI Califo.nia~, Sa Franci.oo. 'IOHN E. CRAIGHEAD. M.D.. Aaaorl.rr Pro/raaor of Porholory. Univenity of Vennonl Medical School. Burlington. T. TIMOTHY CROCKER. M.D.. Pro- /rasor of Mrdreinr. University of Cal4 fornia School of Medicine. San Fran• cisco. CARROLL E. CROtS.S, M.D. Aaa/Hwf ProJrsaor of lnrrrn.l Mrdkinr. Uw'nrer- sily of C.Iifornia School of Mcdicine, Davis. JOHN P. DELANEY. M.D, Pf.D. Aa- aor{arr Pro/rasor of Srf^y. University of Minnesola, Minneapolis. EDWARD F. DOMINO. M.D. Pwlr} aor of Ph.rm.coloty. University of Michigan. Ann Arbor. SP.RTRAM EICHE-t. e3.. D.D3.. DW- rrrro., Institute of Srows.rolork.f )tr- arrrh, Science Resources Foundalioq Walellown, MaM. PROIECT T1T1.E RelaliontN of smolinL ucroids and sncrnlrYScycle so pulmonary c~tonwr- pAoloq awd mucopolysaccharidn CLnges in cytology of IAe tracAcobron• cbial Iree Clsronic bronchitis entNica The dcgr.daMioia, of DDT. TDE and Dicl- dris in sIM cigarette mainstream and aideMreaw rwokes Tlbt anrdi.liow of InRr..rat«T injury of liw. Rok of tRe Mrrwan al.eolar mneropl\aRW in the Erodtrdiot of emphysema SioloRy usd erlopatsie eRects of respir.- Iory awd oneo/enie •irurs in ortlan cultures of Aunman tcspr.o-y Iract lifaw Epilhelial oeR Ir.ndorwaliow and crd- nana Indueiioc of Caype RNA viruses Biologic activity of tof7.cco srnohe co.- demNea ora respraory nwcosa of ro- deMf, enwines and primates in or~an culture; a histologic and autor.dw- ratihic study Pre.eMiow or re.ersal o( rrbnoresal alaln of ret*irNory epitlaelium produocd by ciprettt aeole cddensate and benao- (. ) rynen. ERects of e1lar tt smoke on Use polmo- narF IynspbORtKUIar aysltm TDe In/l,aewoea of amoling and nieotino on circulatiow in the lowcr limb Effects of toD.eeo smoke and nicotine on the central nervous system Metabolic Lterrtlariorsalaipr between to- b.cco aw.ole and the human n.oulA Metabolic and eytophy.iolotic.l chemical Inlerrelallonthips belween tobacco tmoie and the human mouth 8) 65100 G8G8

PRINCIPAI. INVE.STIGATOR OR IntiTtTVf'H)N CARI.TON K. F.RICKSON, P„ D., As- sirranr rrofrasor of 1 her.n.cofnry .n/ Toricology, The University of Kanus School of Pharmacy. Lawrence. WAI.TF.R B. FSSMAN, hr D_ lro/rr- sor of rayrAdoty. Queens Colkge of The City of Ncw York. Flusaing. T1IFODORE N. FINLEY, M D. Drrc- ror of Prl.nonery Rear.rrh LAor.rory, Mount Zson Hotpiral. San Francisco. (Formerty .t The University of New rROlf.(T T1T1.E Mechanism of learning facilitation hy nicotine Studin or nicotine action upon merrory corudidation See I.adrnan, A. ). PRINCIPAI. INV F_STIGATOR OR tNSTITIrfION IIT RFSEARCH INSTITUTF Chicaso. SHIRLEY l.. KAUFFMAN. M.D.. Asso- r..rr rrofrsxrr of rotholot Down. state Medical Cemer, State Lnivertuty ol New York. Brooklyn. Meaieo School of Medicine, Albu4uer- pe.) OARY D. FRIEDMAN. M D. Srn/or 1lrrnlolo F isr Kaiset F ed ti R ("AaraclerisMio ol .mokers and nonvnok- , * ou on p a e- search Instnute, O.kla.d, Cal. ers AARON 1. LADMAN, Pw.D., lroJraso+ .nI Chairman. Drpnrrmrnr of Anuo- H. IIUGH FUDENBFRO, M.D. rro- Iruo. of Afrdklwr. Uwivera~ of Cali- Cdla,ee antibodies in relation to the eti- oloRy of emplry.ema my. The llnivertity ot New Melieo School of Medicine. Albuquerque. (lni- tiated under Theodore N. Fmkr, M.D.) fornia School of Medkine, San Fran- cisco. PAUL S. LARSON, Pw.D, H..t holrs- MURRAY B. GARDNER. M D, Aaao- An epidemiolotical research program on sor of r/bnnacolop, Medical Colkse of Virginia, Richmwrd. cl.rr F•ro/rssor of r.rAoloIty. University of Southern California School ol Medi- eine, 14e Angeles. Ihe etr<.lot y of human cancer DAVID M. OOI.DFNBf'RO, Sc.D. M.D.. Anod.+r rroJrs.or o( l.rAol- Heterotransplanlalion ttudies with human lung cancer Chlr/ GUSTAVE A LAIIREN27 M D or1, Tem'le University Health Sci- ences Center. PbilsdefpAia. . , , . of MrJicinr, Saint ViaorM Ha.pital, Worcester, Mas. (Now N Newtow- N Welksky Hospital, ew1o4 Ma..) LF-ONIDE GOLDSTEIN. D3e. Re- A study of biaekdrie differences between sr.rch Scieniiar, err.r of Rrsrarrh In NrrolorT and rsycA..rrT. New ler- aer. Neurop.yrcAiatric (rutitrNe, Prinoe- lon. 10SEPl1 1. GUARNERI. Pn.D., DYrc- tor, Pr/mon.ry Arrodiolon liborts- tor), Drp.rrwrrnt of Mrlklne, Saint Vincent Hospital. Worcester. Mau. (Iniatcd under Gustave A. laurtnai, MD.) NORMAN W. HEIMSTRA. NM.D. As- s«1.rr rro/raaor of r+7cAolon: DMrc- ro., 1)rirre RrAario. l.il•oe.rory, Uni- veruly of Sour6 Dakola. Verrnillion. nied'me-habituated and non-habituaied orpuisms by use of high energy rbos. phate compounds The eReH of clK.I'rr' emoke on the im- muno"iul nd mctabolic function of alveolar macrophages Effects of smoking on wstained perform- .nee In a .irnulatcd driving test An invesliplion of the relationship be- Iween snwking deprivation and Uress FFecta of smoking on peripheral viwal acuhy RelNlonship between s;+nmaneous anok- rng and mduced mood change I JOSEPH M. LAUWERYNS, M.D.. Pte.D, rrofrswr Or/Inriau fw Mkrauro/Ic Anerom); Chairman. Es/rrLnrnr.f libor.ro.y of C.rflorrlnson.ry and Grnire/ l.rholoty, University of l.eu- ven, Leuven, Belsiurn. RICHARD A. LFRNER, M D., Scripps Cfink and Research Forn/.rlon, La )olla, Cal. CECII.E LFl1CHTENBERGER, Ph.D.. Head. Department of C7mcArmisrry, Swiss Institute for Eaperunental Can- cer Research. lausanne, Swilncrtand. 92 PROJECT TITLE Bioenergelie and endocrine eRects of atresa and their interrelationshrps with The efTeas of tob.cco smoke inhalation EReeta of tobaoeo smoke inhalation on alveolar macrophages and lung sur- faetant in animal models To inveslip te The ptboRenesis of ura I~ane-induc;d lunsadenurnas in mice; .pecilkally tAe idc.lification of the cells Aich give rise to the adcnomas, and The flne slruetural changes in these cells during neoplasia Carernotiren-induced alteralHM/ in the cell cyck of the Type 2.t.colar epabcli.l cell Effect of ciprelte snwkinR on lipids .nd morphology of alveolar lining material and macrop6ages Prep.ration for publication of a book on the biologic atpccts of tobacco and trrwltiwg The possible eAeet of tobacco smoke and wieocine on tuKabk acid rnctabolisrn Subsidy for publkalion of a supplement to the morroRraph, "Tobacco' See Ouarnni, 1. 1. Tbe IyrnpAatics of the lung; their role in ()uid Irantport nd clearance of air- borne particulate matter Studin on penhtenm viral infection A eorreiated histological. cytolo1lical aad c~toeRemica~l study of the tracheoMon- c~ial trp of mice eapo.ed to cigarette arnok. The interrelation between influenfa virus Iwfeetiowa, eapueure to cigarette .n.ote a.d orher faetor• in the dr.rkprnent of Pulmona.y and MonaAral lesions in mtoe 83

PRINCIPAI. INVF4TICATOR (/R INSTITtrfluN 1. P. L,ONO. Pst D. rrohnor of tA.r- n..coloYy, State University of lo.. College of Medicine. lo.• City. Ct-AYTON G. L.OOSLI, Pw.D, M D, HUrlnlr rro/ryrpr of MrIK/nr awJ r.rAoloty. University of SoWhCrn C.1i- forn:a School of Medicine. Loe Angeles KENNNTlI MP.RRIIL LYNCH, MD. SC.O_ I.I.I), CAantrflor and rro/rr r.w f nrrrlrrr of rarAolo*y. Medical Col- kge of South ('voluu4 ( 1.rle.ron. (lw arociaion.ittt Forde A. Mclver, M D.) INES MANDL. Mt.D, A..lw.nr rrv/rr- or of SwrArnrrsrr). Colleqn of Pbys4 ciam and Surwoni Colu..bi. Uriver- uty, Ne. Yor~ (aty. JOHN It. MANHOI D. 11 , D.M D. rro- Jruor .n1 Drrrctor of rarAology an/ Or.f Dwlnorir, New Jersey Colktt of Medicine and Denrerry. Jersey City. MASON RESP.ARCH 1NSTITUTE, Worcesler. M.a DONALD 1. MASSARO. M.D., Ar.ariare rrofruor of Mrlirinr, Owri e Wu*- Inpon Universitr School of Medieix, Washington. D. C. HANS ME1ER. D.V M. St.e Srkwrir, t6e laclarn I d+oraory. B.r Ilubor. Me. MI(.ktrulal(/OICAL ASSOCIATES, INC_ Setleesda. Md. RICHARD l.. NAEYP M D. rro/rl.or .n/ CA.Irman, Drparrnr.nr of r.rAo!- ofy. Pcnnsyl.eni. Srae l)niversity ('oa- kge of MedKine, Hersbey. PROIECT TITLE Comp.rative cytoclremiul, cytological and hislologK11 uudres of early cFlccts of ciftuette smoke (whnle, gas phase, conNduentH) in mice and in I.+suc and organ cultures from mice and humanr Cardiovascvlar effects of nicotine Furtber studies concerning symparbomi- nMic actions of nicotine L.~ lisaue rr.ction. to airborne cbcmi- cal •nd bolotKal agents Fwvko.meaal faclos and putrrwrna.y d.s e..c: I. A.bcJos duM; 11 Arl.rstrn .nd eoureino*en@, viral .nd ebensKnl T!e rok of hereditary clasea.e inl4l+ilor deAekney in the etiology of pulmonary e.npsyse.u Ela.ldytie brvldo.n in the etioloty of pulmonary empaysema Study of the purported relalion+Aip he- l.nn teno\ing and ebanges in human oral lissue !n riro by routine n,icros- copy, drRerenlial paining, and micro- resPirorneter melbds, and furtbrr sta. tiMleJl elaminallorl of t0'o serK1 of data Pretaly In the principal investi- ptor 1 poscseion Mcah.ninl and biological evaluation of mechanical devien for the monitored c~p~u re of eaPecimenl.l animals to lob~oco terwtae rnhalation Compralive racial prevaknce of chronic broechilis Alveolar cells: protein and glycopodein biwynthesi. Pro(dn synthesi. .nd uecretion by tr.- creal muooaa OncoRerelia In the nbbit: genctie +us- ceptibilitr. .er,ical u.narnissbn of vir.e and envirorwneMrA Infiuences fn rMv an/ be riro studies of Ihe influ- enoe of elogenou. rn.terials on activa- tiow of INesN C-type RNA viruses Smoking .nd the pulmonary bt,%od ves- sels: a puamdative, morpMrlurK rtudy 94 PRINCIPAL. tnVF4i1cATOR OR 1NV.TITUTION ALBERT H. NIDFN. M D, rro/raror of Mrdw inr; Dirrcrur, rrln.orr.ry Diu.u Sr, n n. Department of Medicine. Tem- p1 Ilniversity School of Medicine. Pbrladelphia. JOHN W. PARKER. M.D. Aaaoriarr Professor of rerhology. Universky of Southern C.lifwnia School of Mcdi- cine, Los Angeles. MARY STEARNS PARSIIL.EY, Pw.D, Arr/rtan/ rrofru.» of Analow.y /n Ob- rrrrrw and (:ynrcology. College of Physicians and Sur~eon1, Columbia Llnrvcrsily, New Yurt ('/Iy. S. N, PRADIIAN. M D, !M D. rroJrr- aor of rAarnr.colory. Howard Univer- rity College of Medicine, WsAinpon, D. C. i PROCESS AND INSTRUMENTS COR- PORATION, Brooblm N.Y. WILLIAM REOELSON, M.D. rro/rsror and CA.Lrnan. Droarrn.rnr of Mrlkvl OneoloRy, Medical College of Virginia, Ric~mord. ROBERT C. ROSAN, M.D_ Ar.orlur rro/rt.or of Pathology anI rrli.rrict, St. L.oui. University Medical Seboi, St. Louis- CHARLFS L. ROSP, A.M, Srnlor Diac- ror, Veter.nl Admini.tr.tion Outp.tient Clinic. Boaton. IOHN R. ROWI.ANDS, Pa.D., Sa/ Scientist. Southwest Research Inatilute, San Antonio. 7e1. ULRICH H. SCHAP.PP1, M D. DfrMOr of NrasopAarmsroloty, Mrora Ra rearch Institute. Worcater, Maa. PROIECi T1TlE EffecU of cigarette rnoke, drugs and nol- ior» fumes on the terminal airar.ys with special reference to Ihe terminal bronc6ioks Rlech.niuns of suppression of Cclluler im- munuy by carcinogenic Eydroc.rbonr ERect of oorulkueMs of /ob.cco smoke on wormd and malignant bum.n rrs PiraMOry epithelium In vilro Flfect of Iob.oco wnote on normal mwsc lung tiw. Elfect of nicotine on behavior Elfect of nkotine on behavior and its In- ler.ctiow with drup Conrlrvttion of an elperimem.l device for rwo.Moced eapo.ure of rn.ll el- perirneMal animals to tobacco rnoke inhdalioa RES functior4 tumor induction and growth Broeciop.Mnocury dysplasia See Beil, B. A detaikd Ia.eatiRNIon of the natur, of t!e re.ctbn bet.reew bioiogical materi- nl...d .trrwrqlikeerk contamman(ti u.iry ek¢vo. pu.maRnelk reson.nces a.d o*tkal swec(ro.copie (echnirlues ta..Mlptlow of the dlrect rimulatlon of prrys.'aretk nerve termin.ls by nicotin. Nlootiwe r.beWaratlon to ro.tral are.t of tb oe betla: e8ects upon FEU ud .wo.orwk system CTs.rye~ h EPA .nd behavior Induoed .the FroUsIe(ed intrevenrws admin- INratioe of rrnall do.es of nKUtrne in unreslr.incd cats S5

rRlnclrAL InVESTCATOR OR 1N4TITVTNN'( MAURICE S SEOAL, M D, Cl/nk.f hofru.r of MeLclwr, Trfls University School of Medicine: Diircro.. Drparr- wvne of /nA.4rnon TArr.~r• Roston City Ho.piW, ffowon. LUC1O SEVERI. MD, Dii.rcror, hui- rrlr of Ma.bil An.rorwy and Nurol- D..irion of Cancrr Rrsr.rcA, Uni.ersilr of Prrvgia, Perutiss. Italy. 311011 SHISATA, M D_ P+r.D. Aus ci.rr Pro/ruor of PA.rn..coibr~ Ual- versltT d Hawaii School of 1ljedkinr~ Honolulu. ERIK SKINHO/, M.D_ Chief. Drprt- nrrnf of Nrrology. ffitpebicrt Ho.pi- tal, Copenhagen. Denm..tt. GENE M. SMITH. Pw D_ Atalar.wr Pnv- /ruoe of PsTcholory. Hanud Mediasl School, Massacbusctts Oencral Horqi- lal, flawon. LOUI3 A. SOIOFP, M D, Pro/r+sor of Chnkef Mrlicin..nl Chief. Diriilon of CrdioJon. Tcmpirt lJnive.sity Health Scknees Ccnter. fil.delphia. SHELDON C. SOMMERS, M.D. Clini- c,.l Pro(rsnor of PrAo/eRT. CoBeye o1 'PATricjar e.d Swveortrl Cohrmbi. Ueta>rsitr, New York Cit7. SOUTHWEST RESPARCII INSTI- 7lITY, San Anl.,. ... f tL DAV1D M. SP,tIt., 64 0, , Dt.*rW, D+- trtnwr+Y of PMA.+b(7. The dr'ookd.M HspW Center. Rroodlyn, N. Y. CAROLINE eFDFLL T110MA3, M D, ProJrasor t'w.rruau of Mrdiclwr. TM lo\n. Ho}~ins l/.iversNy School of MedKine, ~•IIi...cxt. P*O1ECl' TITt See C6ododt S. Ae aPqro.ch to Ibe study of internal fac- ton in lung carcinorennir influence of \ormones fllood-borwe urcinotcns in moust lung tunqrigenesit 3tudT of .ensiti.ity of vascular liuue to wicoline The .cwle effed of vnokina upon regional cerebral biood fio.w in smokcrs and twasoten; The effect of sonx pbytio• k+rical Miawli upon cerebral bltwd flow i. rwten and nonanwt en T1U retations between smoking and pcr- .orulitf T1a eRed of lobaaco smo\inr on wr- faetruK pecifie fatty aeida, cardiac per- fonnanse and InHabolism of adipose tisaw SqeciRc elfecis of smoking on mTocardial rnecbanie& rnyoeardial metabolism and on entrmca and formed cknKnls of the bloo0 Hod facton in chronic pdesonar) inflam- .ution, empbyscrna and lunr cancer Spetro.ooqk study of effects of tobacco tlnoke constlluenls on rnammalian (is- asn Pwinp.r7 parcnclymal .herationt in an wAop.ffid "nornul' populalion as rc- IMed lo age, aea and environmental factors Tv dpificawe of diRerenl Individual pel- lerlls of circulatory res/fon+c to claar rMle nnokinr 3c.dks or renclie diRerencc+ betwcen osoat:rt and nowrwlen 86 PR I N(:I PA 1. I NI VUSTIC ATOR OR INSTITUTION IAMFS E. P. TOMAN. ht.D., ProJr»a and CAnrm.n of Pl1arrn.coloty. CM- caro Medical School. Institute for Medi- ul Research, Chicago (deceutd). (Con- tinued under Hector C. Sabelli. M.D.) UNIVF.RSITY OF SAN FRANCISCO. San Francisco ELLIOT S. VFSEI L, M.D. Pro/rtsor and Chdrns.n. Drrarrwwnl oJ Pl1wnw- colnn. Pennsrlvanla State ~Jnivertity Collere of Medicine. Hershey. ROMEO A. VIDONE, M.D. Aawclarr Pro/rssor of P.rAo1o{T, Yak Univcr- sily School of Medicine. New Haven. Cono. PETER K. VOOT, PM.D., Pro/ruor of MLroaiolon. University of Washing- lon School of Mcdicine, Sealtk. JOHN V. WFII , M D.. A1al./.M Pro/rt- aor of Mrdi.rnr. University of Colo- r.do Medical ('enter. Denver. SIMON H, WhNDER, PMD., Reararc6 Pro/rrwr of EirxArmirlrf. University of Oklahoma. Norman. T"OMAS C. w'FSTPAI.I. PM.D. Am aori.u Pro/easoe of Plwrnacdop. Uni- verfsty of Vrrrinia Schtol of Medicine. (-harlotlervdte. PROJECT T177.E Studia of P/~c6olo~iul dif/ertncxt be- Iwet~ amoken and nonwnokcn aa aAow~ by comparison of figure draw- iwp P.rchological cAaracteristics of healthy youn4 adults and Ib,eir bioiofical im- PIinllon; a continuinR uudy in depth. wi/h special refercnce to the precvrsors of lypertewsioe and coronary heart di.- eest and to aeoliry habits A study of tbe precvrsors of hrperttrrios and owanary diu..e Cipretle .no\ing p.tttrm over time: ProMn of former a.w\ers eomp.red wit\ continuing sexrt<ers and non- .nokers Medaoiwns of the Psycholropic eRecis of .icotine TV study of actions of nicolinc upon eetMr>rlq n.Pscs and its correlation with the wsecsamsm of actron of behavio.- Nly neti.e drup Induction of srauarnous or11 carcinoma of the lung iw mice Radioirnmuno.aay for nicotine HisloArrnirrry of epithelial muciro in carcinoma of the lung Avian tumor virusts in mammalian bps ERocts o1 cip relte.nolinr and of chronic airway obstruction on hyposic venti- latory drive in man Effecl of cigarette smolc and ils cornpo- nenla on free proline in animal tissue cultures Action of nkdine on aubcellular dislri- bution of ealtcholamines and .erodonrn in brain and heart Influence of nicotine and relaled drup on Iba uplake, MtxaK, rrtcsse and lurno.er of catecMd.mnes in central and peripheral tissue 87

rRINt:IrAL 1NVt<1TtcATOR OR tN3TtTlfrtcN/ rltO)ECi TITLE DANIEL H WISEMAN, M D. Aubr.wr renJnent Pulmon.ry dysrunction follow- lrn/ruor of lrlurrkr. Uni.trutr of inR specdk, lower rtspuao.Y dw.xf SoWhern California. Chilbrw's p/rl- durina csildhc.od .1on, Los Angeles Courwy Gcnerat Hos- pptat, Los Angeks ' Completed Projects Following is a list ot the principal investigators or institutions of c jccb that have been completed prior to the period covered in this port. Several of the individuals named are deceased. The titles and affitiations listed are those in ellect at the tirne the work was completed. tisual• R. FREDERICK DECKER. hID. Aaso- ci.rr rro/taaw of Anuowry .wf Drrc- ror, [iAor.rory of rrrlw.r.l Sclrwcr, Duke Universily Medical Ce+teer, Dw- Aarn, N. C. RALIH S. DECKER, Pit D., hoJt.aor of Che/nWry. Universitr of Houston. Houuon. JOHN A. SEVAN, M.D0. Ro/ruor of rh.n...colol f. llniverdt y of Calitornia School of Medkine, Lo. Awgeks. CI'ZARP. etANCIFIORL M D., D/rldow of Centrr Rruwrh, U.i.ersity of reruaia, rerugia. lialy. CLARENCE M. AGRESS. M.D, Auo- cwr C/iek.l rrofri.or of Mrlklwr. Universiy of Cali(ornJa Medical Ceo- ler, Lo. Angeks. ANTHONY A. At sANFSE, rw.D., DI- rrcro., Nrrrlrbw .wI AIN.AoIk Rt- swth DJr/riow, TM Sutls ReVEitit.- lion CesMer, W6ih rl.in., N. Y. ANTHONY P. AMAROSE, rw.D. Iw- mrnor. Drpiarrnvw/ of Olsrenkt .wf Gyrroologl, TDe Alb.ay Medie~l Cd- kle of Union Uni.erily, Albany. N.Y. E. T. ANOELAKOS, M.D, rr.D. rro- (tuor of rh)dolop, Boston U.i.enMr School of Medicine. Boston. D. MURRAY ANOEVINQ M.D. U.i- .enitr of Wisooesis School ot Medi- ei.e, M.disow. !lROOA A. BARNES. N D., rt D. ho- /ruor (./iflut) of IA'.lofory. Coto- r.do Stale Uei.ersily, Foet Coltiea. FREDERICK W. !lARNL'S, la, M.D. AuocWr rroJtuw of Metklwr. Th. loAn. Hophin. Uui.er.iry School of Medicine. haltinwrc. T. C. BARNES. D.Sc. Rt.r+ec~ Sc1rr,Hu. rbil.6elp6ia St.u Ho.plal, rt+il.del- HYLAN A. ts/CKERMAN, M.D., AuW- .wr Iroj.saor of Afr/krwr, and AL V AN L. !•ARACH, M.D. Cowsr/r.nt !n Afrl- 1Nwr. Cotws.4ia U.i.ersi/y Cdkp of rfrieires awd Swaeon.• Goldw.ler Mesnoeid Ho.Oit.l, New York City. FRED O. !lOCK, rte.D., Aswrl.rr C.ncrr RrarticeA Seirwel.r, S.olork.f Sr.r/on, Roswell Park Memorial Inslilule, S*rielR.itle, N. Y. HERMAN V. /OENIG, rw.D.. Hrd, CArwr/M.y .wf IbrMnrirrrY Drp.rt- n.rwr. SN~elo' Reae.rcb Center, l.et- 1eRso.. ~(r. JAMES F. DONNER, ra.D., t'ro/ruor of •1ologr. Calitor*Ia lnMitule of Tee\- .olorr. ra..dew.. WALTER M. sOOKER, rw.D., rroJer wr and He.f, Drp+rrmrnr of lhrn..- colon, Howard University. W.Ji.a- loR D. C. TOM O. BOWERY. rM.D., Itrkilr Rt.- IIw L.Ionvory, Chtmbrry DrNt- twert, Norti Carotiea Sule Collqc, Ra{dd. OEOFFREY L. SRfNKMAN, M D. As- wci.M rro/ta.or o/ Alrliclnr, Wa)^ne StW U.ivetsity School of Medrcra, DevoN. RAYMOND R. SROWN, rw.D. l.eJt} •or o/ Cl/nkrt Oncdo~~ Uai.mMy of Wi.ooria Medical Scbol, M.diso.. )06P.P aRO7P.K, M.D., Iro/ruw .inI Ch.iws.rr, Dtp.rM.rwr of lncholon. LcKiall Uni.et.ily. RcthkYem. Pa. SUE !lUCKINOHAM. M.D. A.dn.w "eser reli.rrkr, Columbia l)d- twdlf of PAysician. and Srtr- Reowti New Yotk c'ity. sICN)AMIN 0t1RROWS. Ii1 f), A-l.rr rroJrtior of Mrllc/nr, ltnivenuy of Ctacam Chicago. 89 1 89

E M. BUTT, M D. Chief Perhologlir, 1 aa Angeks County General Hosprtal, l oe Angeks. RICHARD U. BYERRIIM, Pst 0_ Pro- /rsur of CArn.irtry, Mk-hip. Stala Unr.enity, Fasl lAnsing. SISTER M. EMILY CAHILL, rwD., Chalrw.en, CArw.lsrry Drpairtn.rnr, Regis College. WeMon. Mm. WTLLtAM ll. CARNPS, MD. U.iv.r- sity of Ula! Colkgs of Medicina. Sak L.k. CNy. MARCUS N. CARROLL, l.. P1rD. CA/rl. D/rldow of P/1orw..cololly. TM arookd.k lloepit.l (-enaee. Idoollyr< N. Y. LF.OPOID R. CERECEDO. Psw D_ Pir.- tasvr of Sloehrrwlstr7 and NMrltb~, niver.ity of Puerb Rioo Sclool of Mcdicine. San l.u. NrILIJAM O. CLARK. PwD. Di-.ctor, Prychopllr+wocvfM RrsercA L.Iarr- tory, Veterans AMnL.istration Horpital. Sepul.eda. Cal. HANS T. CURKP D.Sc ProJruar of /bchrn.lstry, Coluritbia 0.lversity Cof- kAe of Physiciw and Surteoea. New York City. JAY D. COFFMAN, M D. Srction Hr+d, PrrifArrrf Vasrdat, DrPrrn+rwt. Uni- ver.rty Ho.pilal. Bowo.. DANIEI. COHEN. D.V.M. M.P.H_ Ar- slsr.nr ProJroow of Yeteriw.ry E/Idrrn- lofon and P.AIk HwdrA, Uni.adty of Perwyf.ania Sc6ooi a( Veleti..r7 modiclse, P1H.ddPii.. JULIUS H. COMROQ la~ M.D. D1r+c` tor, C.rdlo.+arrfar Rrx.rcA lnn4rtr, UniverutT of Califorala Medic.l Cew- ter, San Franci.co. DEAN M. CONNORS, MD Araorlur Dirrcro., De~ rn.rwr of Llor.roe7 Mrdklnr, S(. A/ary's Hoqittill. Mndi.Lon, W it PIHIIP C(X)PER, MD. (*+/rd hvftr- 0" sor of SrenP .wd Dlrecsae, Surrkd tn Laboratory of (-rRJ.r PAyry/oP), .~ Albert Plndeln C'otte*e d M.dld.e; B Chief. S.rgkel Srr.kr. VMerssts Ad- m minixration Iloqital, Broaa, N.Y. ROBERT l.. CRAIN, PN D. As3r.ront ProJrssar of Socu.loll, l/m.ersity of Ctficago, Chicago. CEC1L E. CR(nS, Research Department. SI losepA flospild, Burbank. Cd. ALBERT DAMON. Pb.D_ MD.. Lec- rrer on Anrhrnpnloly; RefrarcA Auo- ciurlw Mrdkal Anrhropolo[y, Peabody Mueemn, Harvard University, Cam- bridfae, Mau. R. F. DAWSON. Pn D.. ProJrsinc ol Rnr- ry, Columbia University. Nc.+ ~ ~ City. ANDREW S. DIBNP-R, PH D., F.rrrNrr, PrycAsRrxrrA, T1e Age ('emer of New E.Wland, laic., Boro.. RALPH L DORFMAN. PM.D. Dlrrcror of L.Aoraorks, Nroroeter Foundation for Eaperimental Biolop, Sbrewsburr, M ar. JAMES 1. DYAR. Pn.D., Asrist.nt Pro- trs..o. oJ Qlolog), Bellarrnine College. Loui.vilk, Ky. RICHARD H. EARLE, M.D. Chle/, Pal- ..onry Fr+vrran L.Dr+.rory; Auisrant Profraaor of Mrdrelnr. University of C1McaRO. Clitaeo. IOHN M. ECKSTP.IN, M.D.. Aarlsrant Prolrs.or of Internal Aledic~nr, Stde Umver.ily of Iowa College of Medicine. Iowa City. HYMAN ENOFIBERO. M.D, Atrrnd- Inr PAyskien, Cedars of Lebanon Hos- pilal, Lo. Angeka. HENRY 1. ESBER, Pn D.. Reuarch Inr a.rnolor/sr, Maaow Researcb Institute. Woroester, Maa. HANS L FALK. Prn.D. AlJrnct Arsorl- rr Iro/esror. Drp..tmrnt of PatAolotf. U.i.ersily of Southern C.lifornia Sclool of Medicine. Los Angeles. DANA L. FARNSWORTN, M.D_ Hrnry K. Ol(rw Prolessor of Hyr4nr .nd Di- rector o/ Unirrrs/y Health Services, Hrvarrl Univenhy. Cambridte. Ma». FRANK C. FP.ROUSON, l._ M D.. CAalrn.an. Department of PAr+..col- o~ Tkr Albany Medical ('ollrAe of U.~on Unfvrrsity, Albany. N Y. c WI1.1-IAM 1. FISIIBf:IN, M.D.. Chief o/ t.prdemr,dory.('bicatto Bo.rdof Heal/s, Chicago. RUS.SFLI. S. FISHER. M.D_ Univertity of Maryland School of Medicine, Balti- rnore. B. L. FRFEDLANDF.R, M.D. Dirrctor of Cancer Reuarch, Mount jion Hos- Pilal and Medical Center. $~an Fran- cisco. FREDFRIC A. FRENCH. A.S., DYector of Cancer Chrn.orArr.p) RrxarcA, MouM lxrn Ho+pital and Medical Ccn- /cr, San FFranci.co. JACK FRP.l1ND, M,D_ Assistant ProJrs- sor of Ph.rn..refary, Medkal College of Virginia, Rrchmo.d. GRBERT H. FRIEDELI, M.D_ Chief oJ Pathology, St. Vincent Ho.Pi1.t, Worcesler, Mw. ARTHUR FURST, Pw.D_ Director. lnsri- trtr of Chrnncal sio{ory, University of San Francisco. San Fr.ncLco. GEORGE O. GEY, M.D. DDrctor, Frnnry-Howrp Cancer RrsrrrcA Liili- or.rnry; Assoriar ProJrnor of Sr- itrry. The Johns Hoqkins univer.ity School of Medicine, Baltimore (de- ceased ). THOMAS M. OOCKE, M.D., Assorlur Pro/essw of Prrrrntlrr Mrdkinr .nd Cornnrrnlty Health. New Jersey State College of Medicine and DeNisuy, Jersey City. PAUL CrOl.D11ABER, DDS. Ataoc/atr Pro/rssor of Prrlodonrolop, Har.ard Sclool of Dental Medidse, Boston. IRA GORE. M.D., ProJrsaor of r.rhol- op. Boston Univenil~ Stbd of Medi- crne; CAirJ of l.rAo.atory Srrrkv, VNetana Adminjslralkwr Ho.*ital, West Rorbury, Mass. GERTRUDE Y. OOTTSCHALL, PwD. Aulst.wr Professor of lbchrrw/srry, Columbia University Colkp of Myr- cians asd Surfeo.a, New York CMy. A. CI.ARK GRIFFIN. Pti(.D_ Hrwl of 1llorhre.4ny Department. M. D. An- derson Hospital and Tumor Institute. llniversMy of 7 eau Medkal Center. Houston. ARTHUR L. GROSS. M S.. Srnior sio- chrrnia, SourAweu ResearcA Institute, San AsMon.o. 7ee. MORTON 1. GROSSMAN. PH D., M D., Asaxi.rr Clinical ProJrssor of Mrdl- ciwr. University of California Medical Cenler, Los Aneeks. CARL C. ORUHZJT, PM.D., M D_ Auo- cl.tr /n PAyslolory .nd Ph.rm.coloty, University of PennsyIvania Graduate Sc6ool of Medicine. Pfiladelpbia. FRANK e. OUTHRIF, PN D., Pro(n.or, .nd ERNEST HOIX3SON, Pw D_ As- siuaw/ RrswcA Profrtr,n, Drperrmrnr of Ewtonwlojf, No.t6 Carolina Staa Cofkm Rakigl. H. I!. HAAG. M.D., Pro/rssor of PAar- Rk ~7. Medical College of VirAinia. F. J. HADDY, M.D., PN D., Pro/euor .wd CAdrn.an, DrParrn.rnr of Physio!- ot7. University of Oklahoma Medical Center, Oklahoma City. JOSEPH H. HAFKENSCHIEI_ M.D, Mrdk.f Drp.rtn.rnr, Sandoz P6arma- eeuliealR San Francisco. RICHARD 1. HAVEI_ M.D., A,rluanr Pro/ruor of Mrdkinr. Univenity of California Medical Scboof, San Fran- cisco. HERBERT R. HAWTIIORNE, M.D. CArrn..n, Drprtnrrnr of Srrtrry, Uniretsily of /'ewrnylvania Graduate SeAooi of Medieine, PAiladclp6ia. CLARK Nt. HEATH, M.D.. Professor of Mediclne and D4rcror aJ Health Srrr- kYS, Twfts Univenity, h/cdfo.d. Mu. PAULINB HEIZER. PM D. Research As- •ot/ar in Cytology and Cytochrrninry. So. FrRrcisco Institute of Medical Sci- e.oeti San Frvrciaoo. LAWRENCE L HESTP.R. )a _ M.D. Pro- leaser dwf CA.irw.ew. Dtp.rrnwnt of O1qrNlci wd Gynrcol.yy, Medical Colkp of Soutt, Carolina, ('farkMow. EasE Cl1RT1S HOFF. Pw 1), M D, Pro- /rmr .w/ CAdrrw.n, D/rldon of Pry- cAi.mk Research. Medical ('olkge of VirglniR Richmond. m 90 91 CD J IJ

RUS,SEI.I. L. HOLMAN, M D. Louisian. State UniverLty School o/ Medreinc, New Orkans. OLE A. HOLTERMANN, M.D., Rr- srrcA Srirnrur, Lohws/ l~Aor.rry , Univenily of Nodre t)sme, Notre Dame, Ind. FREDDY HOMBtJRGFR, M.D, Prrsl- /rnr .nI Dbrcror, BioResearcii lsti- tute, Inc, Camhrd(te, Mar. ROBERT W. HULt., PtM D.. Professor of diolotkal Sciences. Fio.ida State Unl- versity. T.Ilabs.ee. GEORGE JACOBSON. M.D. Pno/rs.or onf Hr.I. DrP.rrn.rnl of Ra/Iolor) Uni.er.rly of Southerw C.Ilfornia School of Medicine. Los An~ele~ JERRY HART JACOB.SON, MD. D/rr.r ro., Division of f lKrropAysiotory, Ncw York Eye and Ear Inbn.ary, New Yort City. JULIUS H. JACOBSON Il, M D., Aw- ci.rr Professor of Srjr~ .nI Drrrrar of Srrtkd Rr.wrA, V.i.cairy o( Ver- rrroeM Colkge of Medicine. Burlingtow. MURRAY E. IARVIK. PrD. Associate rroJrssor of Pharn.rolon, Alhe.t Ein Nei. Co1kRe of Medicine. Broa>. N. Y. OSWALD R. IONF,, MD. St. LuLe^s Hoqital, New Yort C'it7. ANDREW A. KANDUTXH. PwD. Su/ ScirnrW. TU J.cltao. LaEaraor7. !.r H.rbor, Me. ARNOLD R. KAPI.AN, Ptt.D. Drrctor, 1,.Iar.roq of Alrlkrl Grnrrks, Cle~..~- lasd PrychiMric Inritute and Ha.prtal, Ckveland. HRATCH KASPARIAN. MD.. Assisl.wr Dirrcro., Carllov.ucrl.r 1..1or.torly: lnrrrrcro. In Mrlrcinr, llal.nnann Medial Colkge and Houpital, Phil.- deiphia. ELIHU KATI; ht D, Associate lti'o(rasar of SorwloPy. University of Ctrcyo. C3icap. ANCEL KEYS, PrMD. D/rrc+r, 1.Aor.- ro+y of rhydological Univer- sMy of Minnesore Sclsoel d Publie Health, Minneapolrs. IOSFPH B. KIRSNER. M D, Pro/rreor of IOHN P. MANOS. M D., lnsrrrcror in BERNARD 1 M(LI.ER M D Assistant Mrdkinr. University of Chicatto tich(wA YonL.py and Oorrrrrolory, Medical . , . . Irolrswr, The Daniel Rn&,Rh Insrrrrrr of Medicine. Chicago. College of South Carolina. Chark>ton. of AnrrowrJ, leflerson Medical Colkge, PETER H. KNAPP. M D. Rnrarch lro- CHRISTOPHER M MARTIN. M.D0. As- Philadelphia. /rua of Pr7chi.rry, Boston Univer•uty siaranr Prolrssor of MrArrinr anI Dirrc- . JAMES O MIt.LER PND M.D Pro- School of Medicine. Botton. ror Di.ision of In/ecrio.u Disraus, . , ., , fessor of hychisrry and Rsycholofl: KENNETH P KNUDTSON M Uni- D New lersey State ('olkge of Medicine, Dirrclor Mrnrd Health Rrsrrrh In- . , . , verooty of Washington Medical tichooi. ler•ey City. , srirrrr, University of Michigan. Ann Snttk. CHARLES C. MccARTHUR, P.r.D_ lsy- Arbor. ALVIN 1. KOSAK. Prr D.. Arsociarr Iro- choloRrsr to the Unrrrrsiry Health Srr.- HUGH MONT()OMF RY As.ori- M D /irssor of Chrmisrry, New York l/nirer- krr, Harvard Unwcrsity, Cambridge. . , . .. «r Professor of Melkinr Universit .i(y, New Yort City. Mass. . y of Penrnriv.nia School of Medicine. ROBERT A. KUHN M.D. Associate rro- CHARLFS B. MrCANTS, Prr.D., A1o- P1il.ddP , lrur D/riaion of Nrrosw erjr New rl.u lro/rasor of Sodr, North Carolina , g , /ersey Su1e Cdkp of Mcdrerne lersey Srate College School of Apiculture. P. O7. MONTGOMERY. 1.., M D., rro- , City. R driN. /rs+or of P.rlblogr. University of ' retr Soutt.we.lern Medical ScbooL MAR VIN KII"HNFR, M t) , New York IIENRY C. Mc(Nf I, 1.., M.D., Acting [Hllas, Unr.vsi(y Medical Center. New York Hral, Drprrrnrnr of Pathology. Lori- City. tian. State University School of Medi- Pn.D. M D., DI- GEORGE E. MOORE eine, New Orkans. ~ rrrro., Ro.well Par Mcrnorial Inali- CHARLES W. L..BELLE, PM.D., Artlsr- Iwe, wf.lo, N. Y. anw Iro/rssor of Enrlronnvnrrl H)- HENRY D. McINTOSH. M.D., Professor eIrnr, Dep.rtmt.Y of Preventive Medi- of Mrfkinr and Dwcror, Crliora} KENNETH M. MOSER. M D., Assisranr doc, Jefferson Medical College. Phila- crlr 1LAorrrory, Duke University Professor of Alrlkinr, Geor!<Nown delpisia. Medical Center. Durhum, N.C. Uwl.etsi(y Medical School. Washintt. too D C THOMAS C. LAIPPLY, M D, Professor EDWARD MtKEE, M D.. rro/raror r.f , . . of IrrAolo!<y, Northwestcrn Unirerury Acting CA.irnran, Department of l.- HURLEY LEE MOTI tro/rs- EY M D Medical Sehooi. Chicago. rAoloRy, Medical College of Sowtt Cu- . , . . sor of Afrllclnr and Director Carlio- olins, Charleston. , Rrspirrrory Labrara~ Uni.ersitr o( ROGER K. LARSON. M.D. Chief of Southern California Sdool of Mcdi- Mrlklwr, Fresno County Hospital, KELLY T. McKEE, MD, Associate ho- ciwe Lo. Angeles Pre.nq Cal. /rssor of Melkinr, Medical College of , . South Cardina Cbarkytoa AVERILL A. LIEBOW, M.D.. Chairman. , EDMOND ANTHONY MURPHY. M.D. Drr.rnsrnr of rarholory Yak Uni- HERBERT McKENN1S, )a, Pw.D. r.o- Se.D., Associur rrolessor of Alosr.rls- , New Haven werrt7 School of Medicine Jrssor of rhrnrarolory, Medical Coi- rks raf Mrfkinr. The lohrw HoP~irw , , Corr kge of Virsinia, Richniond. Uni.etsily Sehooi of Medicine, 9ah1- . .nort. (Foenserly at Univcnity of Coto- PSTEN O. LINDSETH, MD.. P>,.D. St. VICTOR A. McKUSICK. MD. holrs- r.do Medical Center. Denver.) loaeyi'. Ho*ital Re.e.rch Laboratory. soe o/ Medicine. The /okwa irr 3t r.r( Misr. University School of Medkine, WILLIAM 3. MURRAY. Sc.D., Srn/or . , wwre. Sras Scknrlu, The Jackson l aboratory, ROBERT H. LINNELL, Pw.D. Aaocl.ur Bar Huhoc, Me. h-olydsor oJ Chrrnlrtry. University of ROSS L. McLEAN. M D, Associate Pro- Ven.o.t, Burlinston. /ruar of Mrlinn., Emory Univeraity DONALD M. PACE, Prr.D., Professor of School of Medicine. Atlanta. t'hyslolop an/ Director. 1nsNrrrr for HP.RBERT L. LOMBARD. M.D., M P.H., Cellular RrsrarcA, University of Ne- Aoiliur, Cancer Re.ench Institwe, WILLIAM F. MtNARY, 1n., Pw.D. As- hr.atR Llncoln. New ErtRland Deaconern Hospital. Boe sori.rr Professor of .(rwon.f, Boston /os. University School of Medicine. Boston. ALBERT B. PALMER. PHD, Assistant Professor. Dr/+.lnrrnr o/ lrycholofy. DONALD B. LOURIA, M D, Atu+clar NEAt. t.. McNIVEN. Pw.D., The Wor- University of Tokdo, Tokdo, O. rro/es.ar of Mr/hrinr, Cornell Univer- ' eeslrr Foundation for F.perimental Bi- ar. .Ny Medied Colkge, New Yurt ( olo*y, Shrew.pury, Mass. ROSE MARIE PANOBORN, BS, MS, Ar.ori.rr )'ro- Ptr D DAVID E MANN Au4r.nr Food Trchnolr.(rsr an/ l~ ., . . lem~' t/ni fruor of IArw~.r.rin~~ 11/1 lA MEYER. hr f)., A r.nrl.rr rro/rs- rrn. Dirparrw.rnr of Fo.r.l Srlrwrr .n/ , Phdadel vetwy School ol Pharmacy u.r of Or•l r.rh„6.ry. University of Trchnolr.er, (lnlversity of Cahfornia. . Pb- Illinois College of txntistry. Chicago. Davis. 92 93

• Ac I FDWARD W. PEIIKAN, MD. CA.1r. JOSEPH H. ROGF.RS, M D., ffoly Name ERNFST SONDHF.IMER, PN.D. Auo- E. D. WARNER, M.D_ ProJr»or of r.- men. Drp.rrmtnr of rhs.w..rology and F.Drrimrnr.l TArr~~rurirs, Bopon Uni- ver.ity Sctrool of r./cdrerne, Boron of lews Hospiral, Gad.den, At.. BENJAMIN A. Rl1BIN, Ptr D, A„Frrmnr r l rietr lro/rssor, oof Slochrml.ny. Cdktte of Foresrry, Sute University of New Yort, Syracuse. rAoto~y S1ae University of Iowa Col- k~e o[ Medicine, Iowa City. SHIELDS OTAKAR 1. POI.LAK, M D.. P11.D. roJeuor of rre rr Hrolrh, Baylor Univeryity College of Medreine, Hous- T. M. SONNEBORN, PM D. DGrfn- WARREN, M.D. DOrctor of l.lo..rarirs, Cancer Research /ruHrrrr, Errcutlrr Drrrrro., Dover Medical Re- eeercA Center, Inc. Dover, Del. lon HEN Y R rr(,hrf Service rro/rssor of ZoolotT, Indi.n. Univer.ity, Bloomington. New Esyl.nd Deaconess Ho.pital, Baa toa. MORRIS POt I.ARD. Psr.D DLvrtor, Lo6rnl L.Aor.tory, Uni.er.lly of Not» Drune Notre D m Ind R 1. USSEK. M D.. F A C.P., rrrs(lrnr, The Runel F'oundati~.n, Inc., Staten Island, N. Y. SAM SOROF, Pn.D_ Hr.I, DePrtmrnt of M.rromolrcrla Chemistry. The In- aitute (or Cancer Rese.rcl, P6N.dcl- BARBARA K. WATSON, PN.D. Asdn- w LrterbloRiar, M.r.dueerte Ots- d H i d• R h . l li , . e, . M. POMFRAT, PwD_ Dlnrser of lofofk+.f Rrsr.rrA, P...den. Foard.- ow (or Mrdic.l Rexvck Prs.dcna, W. C. RUSSELL, M.D., University of Texa. Medical Center, Houston. PETFR F. SAI.ISBURY, M 1), PN 1). phis. ALEXANDER SPOCK, M.D. Andstanr rro/tssor of Pr/iarrirs, Duke Univertlty Medkal CeMCr, Durbenr, N. C. ee o.p t Associ.re, De- esearc /r/n.rN of 1a[YnloJot~ rnI lmmrnol- op, Har..rd Medkal Schooav fto.w., JOHN S. WAUGH, PIS.D. Professor of C .t. HeeI, Iwrrns/rr Trr.tmrnrCrnvr, Saint CArwstury, Mn...clu.Nle Inslitute of H Dow.nl R PRATT-THOMA3 M D Joseph Hrwptd, Burh.nft. ('.1, FRP.DRICK I. STARE, M D_ lro/ruor of Nanlriow Harvard Univ it S h l TecleoloRy, Cambridge. . . , . Professor of LrAolot., MeAiol Cod- PAUL SALTMAN, Pa.D, Aunr.u.r lro- , ers y c oo of Public Hnlth, Bowo.. RICHARD L. WECliTLER, M.D., CNwI- kle of South Cuolin., C1.rkaow. MARTIN S. PROTTI'1., SS_ D.DS_ Chief. Drrarrmrwt ()..f Pathology. /rasor, DrP.rrmrnr of elorArmivey and NawiNoR, Univer:ty of Soutbern Cali- [orsi. School of Medicine, Los Antetes. C. HAROLD STEPFEE, M.D_ Dlrector of l.rbor.ro.lrs, Met6odi.t HoryN.l, Memphis. rr rAy.bfoRfit, Moaltfipe Ho.Piul Institute of Researc4 Pituburgl. RUSSELL W. WE11.ER. M.D., rrholo- Newark Crty HospittaL Newrt, N. /. WALTER REDISCH. M D A»or/rr rro/rssor of Ctlnk.l Mrilkiwr, Ncw Yort University School of Med{dne, atad NYU Researtll Servioe, Goldwaler Memorial Hospital. New York City. HOBART A. REIMANN. M D. ProJes- sor of Mrbrlnr, HaArwn.nn Medicil College and Hoapits4, PfiladelpAla. ROLLAND C. REYNOIDS, M D. As- slsr.nr lro/rssor of r.rAolory. Uwivcr- .it~ of Tetas Sorrtb..eNerw Medical ScAool, D.tt.s. VICTOR RICHARDI, MD. Chief of Srpte.y, Preshyteria. Medical Cent!er. San Fr.ndioco. WILLIS H. RIESEN, Prt.D. Srn/or e/o- rhrmisr, Ll/r Sclrncrs DIrl,Jow, IIT Research Institule. C1inp. (InAialed under A. WeinMocl, Pf,D.) R. H. RIODON, M D_ lro/essor of r.- rholoty. University of Tea.. Medica! srnnclC Galveaon. SYDNEY C. RITTENSERO P+t D_ h+- rssor of s•crrrkr/ory. Jsiiverwy of tberw C.Irfo.wiK ln. Aftele.. BENSON B. ROP M.D_ Assxbtr M1o- )ruo, of Srtrry; CAM/, C.rllac Sr- =tr_ry. l)niveruty of California Scha>a of MedKlne. San f-r.nclKo. )OROEN U. SCHLEGFL. M D., Psr D. JACK P ST1tONO M D Associate tro- rW, Memorial Hospital of CAeaer fro)rsso. and Charrmee, .lrrrion of . . _ i lessor of ratAo1oPy, Loui.iena State Cou.1T, West Clwter, P.. Urolo~~ Drparrmrnr o/ Sr•t( ry Tu- University School of K},dicine New , l.ne U.ivcruty Schoot of Mrdrernc, , Orleans. I DUANE O. WENZEL, Pn D. Professor New Orleans o PArw.rcolq 7. School of Pfarmacy, . MARION B. SUI?.BEROER, M.D. rro- niversit7 of K..r.s, Lawrcnce. ALV1N R 1)t r SCHMIDT P)r D /rssor .n/ Ch.lrns.m, DrPrtmrN of . . , rrc or . of Cowsrfin Med- Tufts University Drrw..rolory and S)/Ailofo~y New FREDERICK E. WHISKIN, MD. C.M_ r, , ford Mass York Univenlty-Beflevue MedicJ Ccn- D/rector, D/rlsdow of HerlrA .nrf rrr- , . Ier New York Cirf. sow.liry Eqdfi6riaw., Tle AR. CeMer , of New EsK1.nq, Ine_ Boston. ISAAC SCHOUR, D.D.S., P)li)., DSc., RENATO TAOIURI, PwD., Associate Dr.w, Collrtr of Drnrisrry, 11nivenity Iro/essor of lsycAofop, Orsd.Me ROOER J. WILLIAMS, PnD. Professor of I llinoiti C.'AicaRo. School of bu.i.ea. AAwi@i.tralow. CArrw{stry: Dlrrcror, Cl.yrow F'o.w- Harvard Uwi.er.ity, borlo§L sbcArn.ici.! lnarlrrrr, The Uni- Research SELTZER Pw D CARL C .enity of Tes.e. Au.tin. . . ., . ology H.r- sic.l ArvlMo Frllor in fA JEROME F. TTIOMAS, PwD_ lro/rssor , D I Maw: Dr- .ard Ut.ivervt CamDridlIe of San/rarT EngUrnistY, Uei.er.ilf o( 1. EDWIN WOOD, M.D. /nqr.rrorr /. y, , H.r.ud School P.rewrrnr o~ Nrrrlrlon California. Berkeley. Mrrlfc/nr, SoMo. UniverrNy School of , of Public F1ealts, Boston. JANET TRAVELL, MD. Asiotirw ho- MeEkitse, losto.. /rswr of Cun(t.l r/1Wns.nofop, Cor• SUMNER WOOD. l.. M.D., Ads+.wr CHARLES E SHERWOOD. M.D., As- nell University Medical CoikIle, New noJrawr of r.tholory, T1e )obne Hop- sw.nr holessor of R.Aiolotr. Univer- York City !fr Uniwsky School of Medlcine, .ity of Roehester School of Medicine . S.Itlnorr. .nd DenliAry, Rochester. N. Y. LIE SHA TSAI, Pn.D_ Rr.rdircA Associ- .rr, Department of r.rholop, YrJe JOHN P. WYATT, M.D. lro/rnor of DAV1D L. S1MON, M.D_ In,nrcro.. Dr- Univertily Scbool of Medicine, New rarAoloil, St. Loui. University Sclool ParrrnxM of )nrrrnrl Mrllcinr, Cincin- Haven. Conn. of Medicia., St. Louis. t..U Oener.l Hoqit.l. Clndnnati. OEOROE W. SMETTQRS, M.D., A,uxl- .rr /n r.rholooly, Northwestern tlnrver- sity Mediul Scfwd, CAicaso Lt1C1t F-SMfIlf. PH D, f•rn/rrr..r of Rlo- rArmistry, Dartmuwh MedK.l tiahoul, H.nover. N. 11. 94 95