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Tliir. COUNCIL Ton ToBAcco RLSCnHCH-U.S.A., INC. July 25, 1973 MEMORANDUM FOR THE FILE CC s RCi9 FWN HK11140013 SUBJECT: Grant 573-BR2 Clayton G. Loosli, M.D., Ph.D. "Effects of Fresh Cigarette Smoke Inhalation the Respiratory Tract of Mice." Site Visit: July 16, 1973. I spent an entire day with Dr. Loosli and his group after I found that Dr. O'Brien was not in town. Included were Dr. K-S Hung, Ph.D., an anatomist trained with Dr. Chapman at Kansas, Mr. Shermans, M.S., a biochemistry graduate student, Mrs. M. S. Hertewach, M.S., and Helgard Niewisch, D.V.M., pathologist. In addition 1 met about 5 technicians and assistants. At this time the survivors of the Sendai infection are being smoked. The new Walton machines were operating, two at a time. With the CD1 mice were on 2 cigarettes, 22 seconds on and 38 off; purging was very rapid, with some smoke coming out past the sponge rubber. Exhaust aperature may not have been large enough. There was no gross mixing failure or deficit. C57 mice can take 3 cigarettes instead of 2. The rooms were clean. In the environmental health unit the caging was not too good, rooms clean but hard to maintain except in gas controlled areas. He is not going to get more Snell mice. He will substitute dba-2 or C3H mice and get advice on AHH levels. He will vaccinate against Sendai for long-term experiments. He will challenge some animals in good isolation units to get a repeat on the first accidental experiment. He reaffirmed his report to Vince last month and reported in the letter to RCH. Dr. Loosli showed me two manuscripts that are published in a monograph and reprints will not be available. They were presented in (1) Utrecht, "Symposium on Aerobiology" and (2) "Germfree Research, Biological Effect of Gnolobiotic Environments." (1) was on "vitamin A intake and pathology of airborne influenza A virus infection." The lesions were similar in mice on the 3 levels of A vitamin when given influenza A virus in aerosol. The epithelial cells of the alveoli showed squamous metaplasia, some residual metaplasia in the A low animals. Viral counts and antibody response were similar in all groups. (2) The second paper described the appearance of the lungs of germ free and con•.rentional mice after exposure to inhalation of an influenza virus. He also described the repair processes. Another manuscript on NO2, S02 and 03 exposed mice subsequently exposed to clouds of lethal or sub-lethal clouds of influenza virus. Viral growth was similar in all groups. Male mice more susceptible than females. Inactive influenza virus vaccinated mice effectively for 4 months.

- Z - NKO1 140014 I saw the sections of the Sendai infected lungs. They were like the slides and photos seen earlier and as reported by SCS. Loosli said that he had seen Dr. Furst's material a few days earlier and was not too certain that they might n'ot be Sondai. Dr. Hung spent some time showing his em. studies of lungs, hyperactive macrophages and lysosomes and the localization of ferritin. The granules rrere r located in pinocytotic vesicles of both type 1 and type 2 alveolar in small numbers but in large amounts in macrophages. The Fe was removed by the macrophages and detectable by Prussian blue by light microscopy. These macrophages persisted for 8 weeks. Mr. Sherman has been doing the vitamin A determinations and the determinations of surfactin. He seemed quite competent. Mrs. Herteweck also works on em. and has been following age changes in smoking mice. The appearance of multiple micro crystals in macrophages of mice showed an increase with age. Their nature unknown. Some seemed to have a membrane around them. She thought they might be protein crystals. All in all, this seems like a good unit with breadth and dedication. Loosli seems to look at every slide. I spoke with Loosli about his letter of resignation from SAB while at luncheon. He means it because he feels that he is a large grantee, has been on a long time and wants to devote more time to his own research. He would enjoy being asked to become a consultant. I told him that I would have been happy to have had a chance to propose this myself. I think it should be formalized. i r WOG: ek