The phenobarbital-type induction of rat liver microsomal monooxygenases by perfluorodecalin

Induction of perfluorodecalin (PFD) of the liver microsomal system of metabolism of xenobiotics has been studied and compared with the inductions by phenobarbital (PB) and 3-methylcholanthrene (MC). It has been shown that PFD increases the content of cytochrome P-450, NADPH-cytochrome c reductase activity. Like PB, PFD induces the activities of benzphetamine-N-demethylase, aldrine-epoxidase, 16 beta-androstendion-hydroxylase. Using specific antibodies against cytochromes P-450b and P-450c (which are the main isoenzymes of cytochrome P-450 in the PB- and MC-microsomes respectively), an immunological identity of the cytochrome P-450 isoforms during PFD and PB induction has been found. According to the rocket immunoelectrophoresis the content of cytochrome P-450 in PFD-microsomes, which is immunologically indistinguishable from P-450b, was approximately 70% of the total cytochrome P-450. Two forms of cytochrome P-450 were isolated from the liver microsomes of PFD-induced rats and purified to homogeneity. A comparison of these forms with cytochromes P-450b and P-450e obtained from the PB-induced rat liver microsomes revealed their similarity in a number of properties, e.g., chromotographic behavior on DEAE-Sephacel column, molecular weight determined by sodium dodecyl sulphate (SDS) electrophoresis in polyacrylamide gel, immunoreactivity, peptide mapping, catalytic activity. The data presented demonstrate that PFD induced in rat liver microsomes the cytochrome P-450 forms whose immunological properties and substrate specificity correspond to those of the PB-type cytochrome P-450. These findings suggest that PFD and PB, which differ in their chemical structure, induce in the rat liver microsomes identical forms of cytochrome P-450.     

In vitro cellular effects of perfluorochemicals correlate with their lipid solubility

Preclinical studies comparing perflubron partial liquid ventilation with conventional mechanical ventilation have indicated that perflubron partial liquid ventilation may exert some anti-inflammatory effects. To assess whether these effects were related to the lipid solubility properties of perflubron rather than to nonspecific biophysical properties of the perfluorocarbon (PFC) liquid phase, we studied the effects of PFCs with varying lipid solubilities on the platelet aggregation response to various procoagulants and the erythrocyte hemolytic response to osmotic stress. In both cases, the degree of the response was directly related to the lipid solubility of the PFC. All the perflubron content of erythrocytes was found to be associated with the membrane compartment. The time to reach a maximum effect on hemolysis with perflubron was relatively slow (2-4 h), which paralleled the time for perflubron to accumulate in erythrocyte membranes. The rate and extent of perflubron partitioning into lecithin liposomes were similar to those of erythrocyte membranes, supporting the hypothesis that perflubron was partitioning into the lipid component of the membranes. Thus some of the potential modulatory effects of perflubron on excessive inflammatory responses that occur during acute lung injury and acute respiratory distress syndrome may be influenced in part by the extent of PFC partitioning into the lipid bilayers of cellular membranes.     

Influence of protein molecule conformation on the character of its interaction with a phospholipid bilayer

The adsorption of lysozyme on mixed phosphatidyl choline-cardiolipin vesicles was studied at pH 4.0 and 6.0. The binding constants at both pH were determined at 0 and 22 degrees C. The presence of maximum on the adsorption isotherm at pH 6.0 was interpreted as an indication of the formation of two types of the protein-lipid complexes. This interpretation was confirmed by electron-microscopic observations. On the other hand, at pH 4.0 only one type of the protein-lipid complex was formed. The lysozyme conformation in solution and in the protein-lipid complexes was studied by circular dichroism. It was found that at acidic pH the lysozyme molecule contains a higher per cent of alpha-helix segments than at neutral pH. As follows from the measurements of lysozyme distribution in two phase systems the increase in alpha-helicity results in the formation of hydrophobic patches on the surface of the protein molecule. The results of the present work and of the previous studies of the interaction of red- and oxy- form of cytochrome C with phospholipid allow the conclusion that for peripheral proteins the nature of protein-lipid interactions is determined by the protein alpha-helix content and by hydrophobic pattern of the protein molecule surface.     

Prediction of the three-dimensional structure of aflatoxin of Aspergillus flavus by homology modelling

Aflatoxins are polyketide-derived secondary metabolites produced by Aspergillus spp. The toxic effects of aflatoxins have adverse consequences for human health and agricultural economics. The aflR gene, a regulatory gene for aflatoxin biosynthesis, encodes a protein containing a zinc-finger DNA-binding motif. AFLR-Protein three-dimensional model was generated using Robetta server. The modeled AFLR-Protein was further optimization and validation using Rampage. In the simulations, we monitored the backbone atoms and the C-α-helix of the modeled protein. The low RMSD and the simulation time indicate that, as expected, the 3D structural model of AFLR-protein represents a stable folding conformation. This study paves the way for generating computer molecular models for proteins whose crystal structures are not available and which would aid in detailed molecular mechanism of inhibition of aflatoxin.     

Solution of a set of Maxwell-Lorentz equations for a ring relativistic electron beam

Nonlinear solutions to a set of Maxwell’s equations and the relativistic equations of electron motion are obtained that describe the equilibrium of a high-power ring relativistic electron beam against the background of immobile ions. By transforming the basic equations, a set of equations for a three-component vortex vector field is derived that describes ring beam configurations for plasma confinement. An example of a numerical calculation of the steady state of a compact beam torus of immobile ions and relativistic electrons is presented.     

Induction of microsomal cytochromes in the rat liver after intravenous administration of emulsions of perfluoro-organic compounds

Intravenous injections of perfluoroorganic emulsions to rats in a dose of 3 ml/kg led to changes in the composition and activity of enzymes of the liver microsomal membrane monooxygenase system. At the peak of induction, i. e., on the 3rd post-injection day, the levels of microsomal cytochromes P-450 and b5 increased 2.8- and 1.9-fold, respectively, as compared to the control. Simultaneously, the rate of NADPH oxidation in the microsomes and the rate of hydroxylation of substrates I and II showed an increase. Conversely, the rate of NADPH-dependent peroxidation of microsomal lipids on the 2nd-4th post-injection days reached its minimal values. These injections stimulated the detoxicating function of rat liver as evidenced from the duration of the hexenal sleep of the animals. All the changes in the monooxygenase system parameters were temporary and reached the control level on the 10th-14th days after injection. It was demonstrated that the main component of the perfluoroorganic emulsions, perfluorodecalin, was responsible for the induction of the monooxygenase system enzymes.