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Groups of male Fischer-344 rats were fed either a choline-supplemented or a choline-devoid (CD) diet, for up to 14 months. In rats fed the CD diet, hepatic lesions developed and progressed through two distinct stages, the first characterized by severe steatosis and an increase in cell turnover and the second by gradual clearance of the deposited fat, fibrosis and parenchymal nodularity. Large hepatocellular carcinomas were found in rats killed at 14 months. DNA was purified from the livers of all groups of rats and from the tumors, and its level of methylation was analyzed using the restriction endonucleases HpaII and MspI. DNA undermethylation was detected only in the livers of rats fed the CD diet for 14 months, whether bearing tumors or not, and in three of four hepatocellular carcinomas. Undermethylation of liver total DNA is therefore a late effect of dietary choline deficiency in the rat.

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A commercially available partially hydrogenated fat was analyzed for fatty acids containing conjugated dienes. The fatty acids were isolated by high-performance liquid chromatography (HPLC), and analyzed with a photodiode array detector and an atmospheric-pressure ionization mass spectrometer. Conventional and second-derivative ultraviolet (UV) spectra of the peaks eluting from the HPLC were recorded with the photodiode array detector, and peaks displaying second-derivative UV spectra characteristic of the conjugated diene chromophore were analyzed by mass spectrometry. The UV and mass spectra of the fatty acids with conjugated dienes, present in the partially hydrogenated fat, were identical to those of reference pre~ parations of linoleic acid isomers with conjugated dienes. The results obtained emphasize that care must be exercised in the interpretation of clinical and experimental data concerning the detection of conjugated dienes in tissues or body fluids of humans and experimental animals. The conjugated dienes may not reflect an ongoing process of lipid peroxidation, but may be of dietary origin.