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Was performed, and thus depletion of the endogenous bile acid pool occurred for 1 h see Rahman & Coleman, 1986 ; , before the addition of CB or saline. Bile and perfusion-fluid samples were collected in preweighed tubes on ice. The volume of bile was determined gravimetrically, a density of 1 g being assumed. The health of the livers was monitored by analysing the extent of leakage into the perfusate of the cytosolic hepatocyte enzyme aspartate aminotransferase EC 2.6.1.1 ; . In no case was the leakage of this enzyme increased by any of the treatments performed. Specific determinations Aspartate aminotransferase was assayed using kits supplied by the Boehringer Corp., based on the method of Bergmeyer et al. 1978 ; . Bile acid concentrations were determined with hydroxysteroid dehydrogenase EC 1.1.1.50 ; as described by Coleman et al. 1979 ; . Phospholipid present in bile was determined by the method of Bartlett 1959 ; , after lipid extraction by the method of Bligh & Dyer 1959 ; . Rat serum albumin in the perfusion fluid and bovine serum albumin in bile were determined by quantitative radial immunodiffusion by the method of Mancini et al. 1965 ; with specific antisera. Authentic rat and bovine serum albumins were used for standardization. 5'-Nucleotidase EC 3.1.3.5 ; and alkaline phosphodiesterase I EC 3.1.4.1 ; activities in bile were assayed as described by Godfrey et al. 1981 ; . Cholesterol was analysed as trimethylsilyl ether derivatives, in order to prevent loss of parent compound in the column, by a slight modification Rahman & Coleman, 1986 ; of the method of Vanlerenberghe & Cassaigne 1968 ; . This method was used i ; because of the small quantity of cholesterol present in rat bile, and ii ; because cholesterol at low concentrations cannot be detected satisfactorily by other methods, owing to interference by other molecules, e.g. bilirubin. Table 3 ppi prophylaxis frequency and risk factors n 214, for instance, anafranil prozac.

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