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TIII: COUNCIL FORTOBACCO RI:SI•:ARCII-U.S.A., IhC. July 22, 1975 MEMURANDUM TOs Drs. Bing, Feldman, Hockett, Sommers and Stoaie FROM: W. U. Gardner ~;E~~1( ~, l~11l43 SUBJECT: Application 814-AR1. Una S. Ryan, Ph.D., Senior Scientist, Papanicolaou Cancer Research Institute, Miami, Florida. "Endocrine Functions of the Lungs." It is difficult to determine just what will be the major activity during the coming year. In general it will be to continue the localization of the site of action of the angiotensin converting enzyme (ACE) in cells and cell organelles. They have found that the same enzyme that acts on angeotensin I to make angiotensin II destroys bradykinin. It seems to be located in the pulmonary endothelial cells and to be associated with the luminal plasma membrane of these cells. Methods have been worked out to obtain and grow endothelial cells in cultures that can be maintained for up to 24 days and ce11s grown in FCS free media ban be followed for secretions into the media. A sensitive assay has been worked out using bradykinin but it is too tedious for practical use. Attempts will be made to improve assay methods. Antibodias to ACE have been prepared and partially purified. Prelirnitr:ary studies of the localization of antibody to antigensin in tissue and cultured cells have been done but questions of artifact remain. They will continue to attempt to improve protocols and reagents for T more precise localization and quantitation. They list some resources that they have available that might be made available to others. Budget. Salary item includes part salaries of three professionals and three tethnicians, one full-time. A smaller amount is for expendable supplies and approximately $4,000 for permanent equipment: camera and still. They list approximately $57,000 available from other sources (NHLI and American Heart). W.U.G. WUG:ek