Quantitative analysis of neurological effects after cranio-caudal gamma-irradiation of rats

A study was made of the frequency and duration of neurological disorders after cranio-caudal irradiation of rats with doses of 7.5-500 Gy delivered to the head. As the mean effective dose increased the neurological disorders occurred in the following order: tremor, rotatory movements hyperkinesia, opisthotonos, and convulsions. The neurophysiological disorders observed were subjected to a pathophysiological analysis.     

ATP synthesis in the membrane fragments of Staphylococcus aureus cells induced by a sudden increase in pH

Membrane fragments of St. aureus are able to synthesize up to 7 nmol/mg protein of ATP after a jump-like increase of pH value. The presence of respiratory substrates is obligatory. The effect is realized only during the jump, if pH passes of 7.4 value. ADP phosphorylation induced by pH-jump is completely inhibited by DCCD.     

Interaction of flavin mononucleotide with dimeric and tetrameric forms of muscle phosphorylase beta.

Interaction of flavin mononucleotide (FMN) with dimeric and tetrameric forms of rabbit muscle glycogen phosphorylase beta has been studied under the conditions when allosteric activator binding sites are saturated by AMP (1 mM AMP; pH 6.8; 17 degrees C). Simultaneous use of schlieren optical system and photoelectric scanning absorption optical system of analytical ultracentrifuge Spinco, model E, makes it possible to register the oligomeric state of the enzyme and calculate the degree of saturation of individual oligomeric enzyme forms by FMN. The apparent association constant for the equilibrium dimer in equilibrium with tetramer decreased with increasing FMN concentration. The microscopic dissociation constants for the complexes of dimeric and tetrameric forms of glycogen phosphorylase beta with FMN have been found to be equal to 10 and 79 microM, respectively.     

Comparative study of the reaction kinetics of cytochrome P-450 reduction by NADPH-cytochrome P-450 reductase and dithionite

The reactions of NADPH- or dithionite-dependent reduction of cytochrome P-450 were studied using a stopped flow technique. It was found that the kinetic curves for both reactions may be fitted by a sum of the two exponents. The arrhenius plots for the fast phase rate constants are linear for both reactions. On the contrary, the breaks on the corresponding plots for the slow phase rate constants are observed at 22 and 33 degrees C for cytochrome P-450 reduction by dithionite and at 31 degrees C for NADPH-dependent reduction of cytochrome P-450. The coincidence of the values of the rate constants and activation energy (56 +/- 5 kJ/mol) for the fast phase of NADPH-dependent reduction of cytochrome P-450 with values of catalytic constants and activation energy for demethylation of tertiary amines suggests that the first electron transfer process from NADPH-cytochrome P-450 reductase to cytochrome P-450 may be the rate-limiting step. A diverse character of the kinetic parameters for the two cytochrome P-450 reduction reactions is indicative of different nature of biphasity of these processes.     

Mechanism of interactions of alpha-naphthoflavone with cytochrome P450 3A4 explored with an engineered enzyme bearing a fluorescent probe

Design of a partially cysteine-depleted C98S/C239S/C377S/C468A cytochrome P450 3A4 mutant designated CYP3A4(C58,C64) allowed site-directed incorporation of thiol-reactive fluorescent probes into alpha-helix A. The site of modification was identified as Cys-64 with the help of CYP3A4(C58) and CYP3A4(C64), each bearing only one accessible cysteine. Changes in the fluorescence of CYP3A4(C58,C64) labeled with 6-(bromoacetyl)-2-(dimethylamino)naphthalene (BADAN), 7-(diethylamino)-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM), or monobromobimane (mBBr) were used to study the interactions with bromocriptine (BCT), 1-pyrenebutanol (1-PB), testosterone (TST), and alpha-naphthoflavone (ANF). Of these substrates only ANF has a specific effect, causing a considerable decrease in fluorescence intensity of BADAN and CPM and increasing the fluorescence of mBBr. This ANF-binding event in the case of the BADAN-modified enzyme is characterized by an S50 of 18.2 +/- 0.7, compared with the value of 2.2 +/- 0.3 for the ANF-induced spin transition, thus revealing an additional low-affinity binding site. Studies of the effect of TST, 1-PB, and BCT on the interactions of ANF monitored by changes in fluorescence of CYP3A4(C58,C64)-BADAN or by the ANF-induced spin transition revealed no competition by these substrates. Investigation of the kinetics of fluorescence increase upon H2O2-dependent heme depletion suggests that labeled CYP3A4(C58,C64) is represented by two conformers, one of which has the fluorescence of the BADAN and CPM labels completely quenched, presumably by photoinduced electron transfer from the neighboring Trp-72 and/or Tyr-68 residues. The binding of ANF to the newly discovered binding site appears to affect the interactions of the label with the above residue(s), thus modulating the fraction of the fluorescent conformer.     

Changes in synapses after gamma-irradiation of the rat head

High reactivity and, at the same time, flexibility of interneuronal contacts were observed after exposure of rat head to 2-100 Gy radiation. At high doses (200-400 Gy) radiation-induced changes played a major role in the development of the cerebral form of radiation sickness. A complete asynapsis is probably one of the causes of the animals death "under the ray" (irradiation of the head with a dose of 1000 Gy).     

The Change in the Linear Energy Transfer of a Clinical Proton Beam in the Presence of Gold Nanoparticles

Biophysics - Gold nanoparticles are promising radiosensitizers for proton radiotherapy. However, the physical mechanisms of gold nanoparticles radiosensitization remain unclear. In the present...     

Molecular organization of the microsomal oxidative system: a new connotation for an old term

The central role that cytochromes P450 play in the metabolism of drugs and other xenobiotics makes these enzymes a major subject for studies of drug disposition, adverse drug effects and drug-drug interactions. Despite tremendous success in elucidating structures and mechanisms of cytochrome P450 function, the concept of the drug-metabolizing ensemble as a functionally integrated system remains undeveloped. However, eukaryotic cells typically possess a multitude of different cytochromes P450 that are co-localized in the membrane of endoplasmic reticulum (ER); they interact with each other through the formation of dynamic heteromeric complexes (mixed oligomers). There has been growing appreciation of the importance of developing an approach to study the ensemble of cytochromes P450 as an integral system inspired growing interest of researchers to the principles of molecular organization of the microsomal monooxygenase system. Academician Archakov and his colleagues made important contributions to this field during the initial period of studies. Subsequent exploration of the molecular organization of the microsomal monooxygenase system as an integral multienzyme and multifunctional system have had an essential impact on our understanding of the key factors that determine the changes in human drug metabolism and other cytochrome P450-related functions in development and aging, as well as under the influence of various pathologies and environmental factors.