Hepatocytes isolated from fed and fasted rats have been used to study the rate of ethanol elimination at different incubation temperatures. In the presence of exogenous pyruvate, hepatocytes from fed and 42 hr fasted rats, eliminated ethanol at 37 degrees by a rate of 11.6 +/- 3.4 and 6.4 +/- 0.8 nmol/min./mg of cell protein (+/- S.D.; n = 5), respectively, which are comparable to the rates obtained in vivo. The ethanol oxidation rate in cells from rats of both nutritional states correlated linearily to the incubation temperature (t = 24-37 degrees) with a temperature coefficient (Q10) of 1.8-2.3. (Q10, (= temperature coefficient) is the factor by which the enzyme activity is increased on raising the temperature 10 degrees). These findings indicate that the oxidation is not controlled by processes which involve membrane transitions in the temperature range 24-37 degrees. Our results indicate that a hypothermic individual with a body temperature of 27 degrees would have a 40-50 per cent inhibition of the ethanol elimination rate. Thus, the observed dependency of the ethanol oxidation on the body temperature has to be regarded in back-calculations of blood ethanol concentrations in forensic toxicology.

Male Wistar rats were given ethanol (approximately 25% of total caloric intake), while two different control groups were pair‐fed isocaloric amounts of lipids or sucrose. After 7‐10 weeks the following organs were studied: liver, cerebrum, heart, diaphragm, kidneys and testes. In fasted, ethanol treated rats there was a reduction in the hepatic concentration of RNA and the cerebral RNA/DNA ratio, when compared to both control groups, while no effects were found with respect to organ weight and amounts of protein, RNA or DNA in heart, diaphragm, kidneys and testes. When fed, ethanol treated animals were compared to both control groups, no effects on organ weight and composition were found in any tissue studied. Several significant differences were registered in the ethanol group as compared to one control group only, as well as between the two control groups. The consumption of ethanol (25% of total calories) thus caused only minor alterations in gross organ composition. These results also indicate the importance of interpreting with care any apparent effect of ethanol ingestion, unless at least two different control groups have been employed.

Hepatocytes were isolated from livers of fasted rats by a two-step Ca+ +-free/collagenase perfusion method. Suspensions of parenchymal liver cells were incubated in the absence and presence of three different anaesthetics, diethyl ether, pentobarbital and fentapyl at various concentrations. Their influence on the hepatocytes was monitored by measuring protein synthesis as the incorporation of L-(UJ4C) valine (50 mCi/ mol, 4.2 mM) into liver proteins. Diethyl ether representing anaesthetics mainly affecting cellular membranes unspecifically, inhibited protein synthesis markedly, concentrations of approximately 10, 20 and 30 mM caused 27, 50 and 74 per cent inhibition respectively, of cellular protein synthesis. The rate of synthesis of medium proteins was somewhat more reduced indicating a greater susceptibility of the synthetic process of these proteins or that ether also inhibited protein secretion from cells to media. The effect of diethyl ether was completely reversible when the anaesthetic was removed by changing the medium. Pentobarbital representing barbiturate anaesthetics, reduced the synthesis of cell and medium proteins very little, while the opiate anaesthetic fentanyl had no inhibitory effect. These results demonstrate a potential hepatotoxic mechanism for membrane active drugs like diethyl ether. They also indicate that special precautions should be taken when this type of anaesthesia is used during the study of hepatic protein synthesis.