Jacqueline F. Sinclair

In primary cultures of human hepatocytes prepared from three separate livers, ethanol increased both CYP3A and CYP2E1, as detected immunochemically. Isopentanol, the major higher chain alcohol in alcoholic beverages, also induced CYP3A and CYP2E1. Maximal increases in these P450s occurred at the lowest concentrations of isopentanol examined, 0.1 mM. Ethanol and isopentanol were each more potent and more effective at inducing CYP3A in the human hepatocytes than was previously shown in cultured rat hepatocytes. Steady-state levels of CYP3A3/4 mRNA were increased by both ethanol and isopentanol. Ethanol and isopentanol induced immunoreactive CYP3A to a greater extent than did phenobarbital. In all three cultures, the increases in CYP3A after treatment with ethanol were less than those observed after treatment with rifampicin, a highly effective inducer of CYP3A in human hepatocytes. In one human hepatocyte culture, the lowest concentration of isopentanol tested increased CYP3A protein to an amount similar to that increased by rifampicin. In another human hepatocyte culture, however, the amount of immunoreactive CYP3A increased by isopentanol was less than that increased by rifampicin. In this latter culture, the steady-state levels of CYP3A3/4 mRNA increased by 0.1 mM isopentanol and 1 microM rifampicin were similar. This is the first finding of induction of CYP3A in human hepatocytes by ethanol or isopentanol. The clinical significance of the findings is discussed.

We investigated the protein composition of the lipid-containing bacteriophage phi 6. We also studied the synthesis of phage-specific proteins in the host bacterium Pseudomonas phaseolicola HB10Y. The virion was found to contain 10 proteins of the following molecular weights: P1, 93,000; P2, 88,000; P3, 84,000; P4, 36,800; P5, 24,000; P6, 21,000; P7, 19,900; P8, 10,500; P9, 8,700; and P10, less than 6,000. Proteins P3, P9, and P10 were completely extracted from the virion with 1% Triton X-100. Protein P6 was partially extracted. Proteins P8 and P9 were purified by column chromatography. The amino acid composition of P9 was determined and was found to lack methionine. Labeling of viral proteins with [35S]methionine in infected cells indicated that proteins P5, P9, P10, and P11 lacked methionine. Treatment of host cells with UV light before infection allowed the synthesis of P1, P2, P4, and P7; however, the extent of viral protein synthesis fell off exponentially with increasing delay time between irradiation and infection. Treatment of host cells with rifampin during infection allowed preferential synthesis of viral proteins, but the extent of synthesis also fell off exponentially with increasing delay time between the addition of rifampin and the addition of radioactive amino acids. All of the virion proteins were seen in gels prepared from rifampin-treated infected cells. In addition, two proteins, P11 and P12, were observed; their molecular weights were 25,200 and 20,100, respectively. Proteins P1, P2, P4, and P7 were synthesized early, whereas the rest began to increase at 45 min post-infection.

Seizures may occur in acute intermittent porphyria or other hepatic porphyrias. Management is difficult, because barbiturates and hydantoins exacerbate the porphyric state. We studied one patient with major motor seizures and acute intermittent porphyria. The seizure disorder was exacerbated by phenytoin and did not respond to a high-carbohydrate diet or to intravenous hematin. Clonazepam was ineffective in treating the seizures and, in high doses, seemed to exacerbate the porphyria. Both clonazepam and valproate were porphyrinogenic in experimental test systems. Because both drugs may exacerbate the acute hepatic porphyrias, bromide remains the drug of choice to treat these seizures.