Ethanol is remarkable for the wide range of structural and functional changes induced in the liver. The metabolic changes are manifest as fatty infiltration, alcoholic hepatitis, cirrhosis, and fibrosis. These are attributable in part to the direct effects of ethanol upon the redox state of the cytosolic and mitochondrial components which in turn can impact upon major pathways of metabolism. Of great interest is the effect of ethanol upon the phosphorylation potential, a topic which can be investigated in great detail in this project because of the availability of transfected creatine kinase in the liver of the mice under study. Thus, this transfection has made it possible to compare metabolic control by inorganic phosphate (Pi) with control by adenosine diphosphate (ADP). A comparison of the response of these different controls to imposed stresses offer new opportunities to evaluate the metabolic effects of ethanol, free radicals and metabolic overload. At the same time, we will explore metabolic control in a system in which the phosphate potential and hence the thermodynamic properties have been altered ten fold by the transfection. A series of stresses are described, particularly that of the redox state by ethanol. A phosphorus nuclear magnetic resonance spectroscopy (31P NMR) and analytical biochemistry study of the PCr/Pi ratio and the pH values of perfused mouse liver over a range of metabolic and redox stresses will be conducted to evaluate the range of ADP control. Steady state and equilibrium values of substrate-couples, especially ethanol/aldehyde that control PP will be evaluated by 31P NMR, analytical biochemistry and freeze-trap redox studies of the flavoprotein/NADH (FP/NADH) ratio, especially ethanol. Natural or enhanced Chemiluminescence (CL) of radical oxidizing species (ROS) will be correlated with the redox state of the liver, and perturbed by ornithine stress. T-butyl-peroxide stress and alcohol stresses will be evaluated by 3-D freeze-trapped redox scanning. Resistance to stress will be evaluated by ROS and histopathological measurements.
