Effects of chronic neuroticization on the monoaminergic systems of different structures in the brains of rats with different typological characteristics.

The levels of monoamines and their metabolites were studied by HPLC with electrochemical detection in homogenates of hypothalamus, hippocampus, prefrontal cortex, and amygdala in intact and neuroticized Wistar rats with different types of behavior in the open field and forced swimming tests. Intact rats with intermediate levels of activity and depressivity had higher serotonin concentrations in the hypothalamus and lower noradrenaline and hydroxyindoleacetic acid levels in the hippocampus than rats characterized by low activity and high depressivity. In neuroticization, the levels of study monoamines and their metabolites decreased in all the brain structures investigated with the exceptions of an increase in the dopamine concentration in the hippocampus and the dihydroxyphenylacetic acid concentration in the prefrontal cortex. The effect of neuroticization on the neurotransmitter systems in all study structures except the hypothalamus depended on the typological characteristics of the rats. This was most marked in rats with the extreme types of behavior--active and passive--in which changes in monoamine and metabolite contents were seen in all brain structures studied. Rats of the intermediate type showed no changes in any of the substances studied in the hippocampus.

[The influence of chronic neurotization on the monoaminergic systems of various brain structures in rats with various typological characteristics].

Typological behavioral features of Wistar rats were tested in the open field and in Porsolt test. Rats were assigned to groups with high (HAct), medium (MAct), and low (LAct) behavioral activities. The same rats were assigned to high (HDep), medium (MDep) and low depressive (LDep) groups. The release of norepinephrine, dopamine, serotonin and their metabolites in homogenates obtained from the hypothalamus, hippocampus, frontal cortex and amygdala was assessed by microdialysis and HPLC. In these groups, the monoamine concentrations were different: the level of serotonin was higher in the hypothalamus and norepinephrine and 5-HIAA levels were lower in the hippocampus of MAct - MDep rats as compared to LAct - HDep. Chronic neurotization caused changes in monoamine concentrations in the hypothalamus and amygdala in rats of all groups, whereas in the hippocampus and frontal cortex monoamine changes were observed in HAct - LDep and LAct -HDep rats. The most prominent changes in monoamines levels in neurotized rats with different types of behavior were found in the frontal cortex, amygdala and hippocampus. The results show a correlation between the typological of behavioral characteristics and the reaction to stress of monoaminergic systems of the hypothalamus, hippocampus, frontal cortex and amygdala.

[Characteristics of hybrid genes coding functionally-active secretory metalloproteinases from bacilli]. / Kharakteristika gibridnykh genov, kodiruiushchikh funktsional'no- aktivnye sekretornye metalloproteinazy batsill.

A set of different hybrid genes encoding functionally active enzymes was obtained by homologous recombination between the fragments of related Bacillus amyloliquefaciens and Bacillus brevis metalloprotease genes cloned in plasmid vector in tandem orientation. The nucleotide sequences of hybrid genes were analyzed. It was demonstrated that the presence even of short homologous regions is sufficient for effective recombination in Bacillus cells.

[Effect of nucleotide anti-aggregants (NAD, AMP) and ischemia on the tissue blood coagulation factors]. / O deistvii nukleotidnykh antiagregantov (NAD, AMP) i ishemii na tkanevye faktory gemokoaguliatsii.

It was shown in experiments on white rats that combined administration of nucleotide antiaggregants NAD+ and AMP, and NAD+ decomposition inhibitor--nicotinamide, decreased the increment of procoagulant (thromboplastic) activity of membrane particles in the ischemized kidneys and reduced thromboplastic activity of membrane particles in intact kidneys. When nucleotide antiaggregants were added to rabbit citrate plasma, the inhibition by membrane activation of recalcified plasma coagulation was more pronounced with the use of tissue thromboplastin of the kidneys and brain--proteophospholipid liposomes, than with the use of phospholipid liposomes obtained from thromboplastin. The data presented have evidenced an indirect anticoagulant effect of nucleotide aggregants at the level of tissue coagulation factors, and their direct anticoagulant action at the level of membrane activation of plasma factors.

[Effect of nucleotide aggregants and anti-aggregants on the coagulation potential of blood plasma]. / O vliianii nukleotidnykh agregantov i antiagregantov na koaguliatsionnyi potentsial plazmy krovi.

The authors studied the influence of adenine and pyridine nucleotides on platelet aggregation and on the time of calcium-induced coagulation of citrate plasma, rich or poor in platelets, with deficiency of membrane fragments and its compensation with exogenous small-dispersed tissue thromboplastin. Calcium ions stimulated the conversion of small-dispersed inactive thromboplastin into a larger and more active substance. The antiaggregation agents AMP and NAD+ inhibited inclusion of phospholipid membrane fragments into the process of recalcified plasma coagulation, i.e. expressed an anticoagulant effect, while the aggregation agents (ADP, NADH) intensified their procoagulant action.

[Early reversible changes in the glycolytic system of rat tissues in ischemia]. / Rannie obratimye izmeneniia glikoliticheskoi sistemy tkanei krys pri ishemii.

It is shown that the glycolytic system obtained from the ischemically damaged tissues of rats in the process of the long-term functioning in vitro: partial--after long-term (1.5-2 h) ischemia and completely--after short-term (15-30 min) ischemia. Detection of reversible changes in the glycolytic system under ischemia, besides determination of its activity with the short-term functioning is promoted by isolation of the glycolytic system from tissues as well as prevention of the damage in vitro.

[Glycolysis in the rat kidney shortly after ischemia and administration of calmodulin inhibitors, AMP and NAD]. / Glikoliz v pochkakh krys v rannii period posle ishemii i pri vvedenii ingibitorov kal'modulina, AMP i NAD.

It is shown in experiments on rats that the early postischemic period after 1- and 1.5-hour ischemia of kidneys is characterized by a decrease in the damage of the glycolytic system site which induces glucose-6-phosphate transformation into lactate and by an increase in the inhibition intensity of the initial hexokinase reaction of glycolysis. In the postischemic period after more prolonged (2-, 3-hour) ischemia the damage of the glycolytic system develops also at the site of glucose-6-phosphate transformation into lactate. Administration either of the nucleotide complex (NAD and AMP) or calmodulin inhibitors (aminazine and zinc sulphate) to rats prior to two-hour occlusion of kidneys vessels promotes a decrease in the inhibition of the glycolytic system activity in the postischemic period. At the same time the separate and combined application of zinc sulphate and triftazin (the most intensive calmodulin inhibitor) is not efficient. The positive effect of NAD, AMP and aminazine on the state of the glycolytic kidney system in the postischemic period correlates with the improvement of the blood microcirculation processes in them.

[Changes in and pharmacological correction of oxidative phosphorylation in regional kidney ischemia]. / Izmeneniia i farmakologicheskaia korrektsiia okislitel'nogo fosforilirovaniia pri regionarnoi ishemii pochek.

Respiratory and phosphorylating capacities of kidney mitochondria were distinctly decreased within early (1.5 hr) and late (1 day) postischemic periods after long-term (2-3 hrs) ischemia of rat kidney. Preadministration of adenine (ADP, AMP) and pyridine (NAD) nucleotides into the animals prevented the decreases, while vitamin E, heparin, trifluoroperazine or aminazine proved to be ineffective. Depletion of mitochondrial nucleotide pool, which occurred during long-term ischemia of kidney and were maintained within the post-ischemic period, appears to be responsible for impairment of oxidative phosphorylation.

[Effect of NAD and ADP on glycolysis in the kidneys and liver of rats during ischemia]. / Vliianie NAD i ADP na glikoliz v pochkakh i pecheni krys pri ishemii.

Experiments on albino rats have shown that kidney ischemia and its simulation by the anaerobic incubation of postmitochondrial kidney homogenate fraction without a substrate induce a considerable damage of the glycolytic system at the stage of the glucoso-6-phosphate transformation into fructoso-1.6-diphosphate and a less pronounced damage in the fructoso-1.6-diphosphate transformation into lactate. Administration of adenosine diphosphate (ADP) and nicotinamide adenine dinucleotide (NAD) to rats before kidney vessel occlusion or their addition to the postmitochondrial fraction before the anaerobic incubation without a substrate decreased a degree of the glycolytic system damage. The damage of the glycolytic system and protective action of NAD are also detected under simulation of liver ischemia. Possible mechanisms of the ischemic damage in the glycolytic liver and kidney tissue system are discussed.

[Oxidative phosphorylation in pulmonary mitochondria after massive blood loss and restitution by autologous transfusion]. / Okislitel'noe fosforilirovanie v mitokhondriiakh legkikh pri massivnoi krovopotere i vospolnenii ee autokrov'iu.

A procedure is described for isolation and polarographic study of rabbit lung mitochondria exhibiting high respiratory control as well as coupling of oxidation and phosphorylation approaching the theoretical value. The procedure enabled to detect a reversible impairment of respiratory and phosphorylating functions of lung mitochondria under conditions of haemorrhagic shock, which was the most distinct within the first hours after blood reinfusion. Correlation was established between dynamics of alterations in mitochondrial energy production in lung tissue within posthaemorrhagic period and structure-functional characteristics of the lung surface system.

[Comparative properties of mitochondrial and microsomal NAD(P)H-dependent lipid peroxidation]. / Sravnitel'naia kharakteristika mitokhondrial'nogo i mikrosomnogo NAD(P) H-zavisimogo pereokisleniia lipidov.

It was shown that rat liver mitochondria and microsomes contain NADPH- and NADH-dependent systems of enzymatic lipid peroxidation. Activation of the NAD(P)H-dependent process in mitochondria occurs due to inhibition of membrane binding of Fe3+ and Fe2+ by chelators, since the membrane-bound ions of Fe cannot effectively participate in peroxidation. The activity of enzymatic mitochondrial lipid peroxidation decreases at alkaline values of pH. The Michaelis constant for chelated Fe3+ ions under NAD(P)H-dependent mitochondrial peroxidation is by 1,5 order of magnitude more than for microsomal peroxidation. In terms of the chelators' effect on Fe3+ binding by microsomes and peroxidation, as well as pH and Km for Fe3+ the microsomal NADHL-dependent system of lipid peroxidation is similar to mitochondrial NADPH-systems; however, it differs from the latter in the type of dependence of peroxidation on the amount of membranes. Based on the ability of the NADPH-system of microsomes to produce the superoxide radical which participates in initiation of peroxidation together with Fe ions, it is assumed that the decrease of Km for Fe3+ at NAD(P)H-dependent lipid peroxidation in mitochondria and the NADH-process in microsomes can occur in the presence of exogenous sources of active oxygen production.

[Zinc- and copper-containing superoxide dismutase activity in rat tissues normally and in hypoxia]. / Aktivnost' tsink- i med'soderzhashchei superoksiddismutazy v tkaniakh krys v norme i pri gipoksii.

Acute hypoxic hypoxia caused a distinct decrease in activity of Zn2+, Cu2+-containing superoxide dismutase in cytosols of rat liver and brain tissues and less distinct--in liver mitochondria and microsomes, brain mitochondria and blood plasma. On the basis of the data published in the literature, hypoxic inhibition of the enzyme appears to be the significant reason for the posthypoxic activation of synglete-oxygen pathway of lipid peroxidation in tissues, enriched with catalase [liver, kidney, blood ] as well as of hydroxyl-radical pathway of peroxidation in tissues with the low catalase activity [brain, heart, lungs].

[Quantitative method of determining the activity of zinc- and copper-dependent superoxide dismutase in biological material]. / Kolichestvennyi metod opredeleniia aktivnosti tsink-, med'zavissimoi superoksiddismutazy v biologicheskom materiale.

The activity of superhydroxide dismutase was estimated by following rate of inhibition of non-enzymatic reduction of tetrazolium salt in reaction with superhydroxide ions, which were continuously generated in a photochemical system. The quantitative estimations were achieved by using para-nitrotetrazolium chloride, which was reduced to formazanes soluble in acetone. Formation of formazanes by the photochemical system (riboflavin + tetramethylethylene diamine + O2 + para-nitrotetrazolium chloride + light) in presence of biological material and cyanide was compared with formation of formazanes in absence of cyanide for specific estimation of Zn, Cu-dependent forms of superhydroxide dismutase. The rate of enzymatic cyanide sensitive inhibition of the photochemical formation of formazanes from paranitrotetrazolium chloride was determined by the activity of Zn, Cu-dependent superhydroxide dismutase in biological material and might be expressed in units of the activity -- un. ac. = 10/(0,026.% inhibition--1,3).

[Respiratory phosphorylation in the liver mitochondria of rats with transplantable sarcomas]. / Dykhatel'noe fosforilirovanie v mitokhondriiakh pecheni krys s perevivaemymi sarkomami

The method of polarography was used to study respiration and phosphorylation capacity of the hepatic mitochondria in rats with transplantable sarcoma 45 and Walker carcinosarcoma in different terms after transplantation, and in spontaneous regression of the tumors involved, as well. In the hepatic mitochondria of tumor-bearing animals a marked dissociation of respiration and phosphorylation is observed, being mostly manifested in oxidation of glutamate and endogenic NAD-dependent substrates. Non-phosphorylating oxidation suffers to a less extent. Experiments with the use of rhothenon indicated that an impaired rate of the coupling of oxidation and phosphorylation was also related with a destroyed succinate site in the respiratory chain. In the presence of serum albumin an up to normal values restoration of the rate of respiration and phosphorylation coupling is observed in the hepatic mitochondria of tumor-bearing animals. The degree of interference in the respiratory phosphorylation process is mostly significant at late stages of tumors development and sharply declines in their spontaneous regression.