sample="quota" bates="MNAT00558527" isource="atc" decade="1960" class="ui" date="19660615" CONFIDENTIAL The Council For Tobacco Research, U.S.A. June 15, 1966 CTR GRANT No. 346 CONFIDENTIAL PROGRESS REPORT - 1965-1966 ANTHONY A. ALBANESE, PH.D. Director Nutrition and Metabolic Research Division The Winifred Masterson Burke Relief Foundation White Plains, New York EFFECTS OF NICOTINE ON PROTEIN AND AMINO ACID METABOLISM IN HUMANS Consideration of the available information has led to the speculation that a segment of the population may be hypersensitive to the products of tobacco smoke, and that the approach of mass abstention is unsound and not likely to succeed. On this premise, we set ourselves to the task of establishing criteria of such hypersensitivity which might enable physicians and health officers to make individual recommendations on smoking habits. Prior investigations in this laboratory supported by the Council for Tobacco Research - U.S.A., indicated that cigarette smoking alters the utilization of proteins primarily via the metabolism of the aromatic amino acids, and transaminase enzyme levels . Investigations were therefore initiated to ascertain differences in the utilization of the aromatic acids which might be associated with cigarette smoking. PROCEDURES AND METHODS In the screening studies (Phase I), spot and 24-hour urine collections were tested for indigo red by the method described by Albanese . In Phase II studies, 24-hour urine collections were obtained on 3 successive days from subjects on self-selected diets who had been given: Day I, no amino acid load; Day II, 1 gm, L-tryptophan; and Day III, 1 gm. L-phenylalanine. Suitable aliquots were removed from the collections for analyses of indole metabolites. Qualitative and quantitative determinations were made for indigo red content. Indole acetic and indole propionic acids were estimated by extraction and chromatographic procedures recently developed in this laboratory by Louise A. Orto . RESULTS Indigo red tests on morning urine specimens of adult male and female subjects on normal diets revealed no traceable correlations to smoking habits (Table I). (Insert Table I) In a subsequent assay, indigo red measurements were made on 24-hour urine samples collected on the control day and on the following day when 1 gm. of DL-tryptophan was given by mouth. It is apparent from the indigo red readings obtained (Table II, Column b-a) that prior or present cigarette smoking was associated with an increased quantitative production or excretion of indigo red. It appears from these (Insert Table II) results that the indigo red test on 24-hour urine collections may prove useful in assessing the relative effects of smoking on the metabolism of tryptophan. In the second phase of this study, we concerned ourselves with a chromatographic quantitation of the metabolites of L-tryptophan and L-phenylalanine. The reproducibility and accuracy of the procedure are indicated by the data collected in Table III. (Insert Table III) Increases in the indolepropionic acid area with administration of the amino acids were greater in the smokers, M.P. and R.W. than in M.H., a non-smoker, and B.J., the heavy smoker who stopped some 3 years ago. These data are collected in Table IV. (Insert Table IV) SUMMARY: Considerable time was spent in developing and validating the methods and procedures employed in this study. Preliminary results of Phase I suggest that the indigo red test of 24-hour urine collections may prove useful in screening metabolic abnormalities which are associated with smoking. It seems possible that this test may be helpful in detecting individuals who are hypersensitive to the products of tobacco smoke. The data obtained in Phase II encourages a continuation of our line of attack on the metabolic efforts of smoking. The need and desirability for extending these investigations is apparent. REFERENCES Albanese, A. A.: Confidential Report on the Effects of Nicotine on Protein and Amino Acid Metabolism in Humans. 1962-1965 Albanese, A. A., and Frankston, J. E.: A Difference in the Metabolism of L- and DL-tryptophan in the Human. J. Biol. Chem. 155: 101-108 (1944). Albanese, A. A., and Orto, L. A.: Unpublished Data.