Project Title: 14, Scope: cells, Biological Studies of Materials Arising from Energy Operations The Effects of Pollutants on Cells Studied in Tissue Cultures RX-03-06-(c) (Cont'd.) human alveolar macrophages, embryonic human tissue cultures, lymphocyte cultures, time, These tissue cultures would be measured for: 2) growth fraction, 3) and 1) doubling time parameters of che cell cycle utilizing the flow of tritiated thymidine labeled cells through mitosis, 4) the possible induction of somatic mutations (enzymatic changes), 5) changes in function, e.g,, effects on production of erythropoietin and other enzymes by cultures of human renal glomeruli, and 6) cytogenic effects. One theory on the development of emphysema suggests that a result of pulmonary irritants is the interstitial death of neutrophils and/or macro- phages and the concomitant release of potent destructive lysosomal enzymes that produce microscopic foci of necrosis which, over a prolonged period, produce emphysema (Janoff). However, the detection of such an effect is almost impossible by ordinary serial section of lungs because the sizes of the lesions are too small, It is therefore proposed that a tissue culture system be developed to assay such effects, The system would consist of three Layers of different cells in soft agar: the deepest layer consisting of an appropriate feeder system; the next layer consisting of human pulmonary epithelium obtained from viable fetuses; and when this culture is established, a third layer consisting of human alveolar macrophages and neutrophils exposed to particulate atmospheric and/or gaseous pollutants, The basis for this system is as follows: if the phagocytes ingest particles for which the q cell is not programmed to inactivate, there is a high probability that the result would be the death of the neutrophil or macrophage and the release of the lysosomal enzymes (hydrolytic and oxidative) thus leading to the death of underlying pulmonary epithelial cells, Im principle, this could be identified by the development of transparent plaques, If the tissue culture studies produce positive results, . chronic inhalation studies would be initiated using animals exposed to variable concentrations of the same materials used in the tissue culture studies. Lifetime observation of these animals would include: 1) the incidence of pulmonary disease, 2) somatic effects, 3) pulmonary function, 4) reproductive capability, 5) the induction of dominant lethals, 6) the effects On embryo in culture, and 7) carcinogenesis, The proposed studies would be in collaboration with the BNL Department of Applied Science where pollutants would be prepared for use in tissue ggnd in animal studies. Dr, Michael Bender will participate for the M@eic studies, and a scientist will be required for the study of the embryos, (See Continuation Sheet) RX-391 hb 149334 iAfb