Slightly depressed growth and fatty livers have been reported in rats fed moderate amounts of nicotinamide, together with a low protein diet (Handler & Danm, 1942; Handler & Bemheimn, 1943). The fatty livers were susceptible to the action of choline, while methionine was found to alleviate the growth inhibition. Since excess of nicotinamide is detoxicated and eliminated as urinary N1-methylnicotinamide, the observed toxicity of nicotinamide at unphysiological levels of feeding was attributed to forced methylation and consequent depletion of labile methyl reserves (Handler, 1944). In agreement with this, Handler observed no signs of toxicity as a result of ingestion of large quantities of nicotinamide by young rabbits or guinea pigs, neither of which species excrete NL-methylnicotinamide (Handler, 1944). Production of fatty livers by feeding excessive amounts of glycocyamine, cystine or homocystine have also been reported (du Vigneaud, Chandler, Moyer & Keppel, 1939; du Vigneaud, Chandler, Cohn & Brown, 1940; Stetten & Grail, 1942). More recently, it has been shown that, in the pyridine-fed rat, there is a decreased urinary excretion of creatine owing to a reduction of available methyl groups, pyridine being detoxi-cated as N-methylpyridinium hydroxide (Dinning, Keith, Parsons & Day, 1950). The known mediation of pteroylglutamic acid in biological methylations including the methylation of nicotinamide (Fatterpaker, Marfatia & Sreenivasan, 1951, 1952a, 1954; Dietrich, Monson & Elvehjem, 1952) suggested the present studies on its influence on the extent of depletion of methyl reserves in rat livers consequent on ingestion of excessive amounts of nicotinamide. The studies included observations on a number of other substances and enzymes expected to be influenced by changes in pteroylglutamic acid or available labile methyl compounds. Pteroylglutamic acid deficiency was induced in the first set of experiments by dietary means, while in the second series aminopterin feeding was used. In the first series, additional groups with choline supplementation were included. EXPERIMENTAL Methods Urinary excretions of creatine (Clark & Thompson, 1949), Nl-methylnicotinamide (Huff & Perlzweig, 1947) and total nicotinic acid (Swaminathan, 1946), liver N (Johnson, 1941), This work was aided by a grant from the Indian Council of Medical Research. Our thanks are due to Lederle Laboratories Inc. (New York), Chas. Pfizer and Co. Inc. (New York), Glaxo Laboratories Ltd. (Greenford), and Boots Pure Drug Co. Ltd. (Nottingham) for valuable gifts of chemicals.