[Carbohydrate and nitrogenous metabolism condition in the rat tissue under experimental rhabdomyolysis].

Some effects of glycerol injection on indices of the condition of the thiol-disulfide system as well as carbohydrate and nitrogen metabolism in rats in vivo were studied. A decrease was revealed in levels of non-protein SH-groups in the liver, kidney and heart, as well as of protein SH-groups in the kidney and heart of rats following glycerol injection. That might be connected with SH-group oxidation under the excessive arrival of free haem into tissues under rhabdomyolysis. A decrease in glycogen and increase in tyrosine aminotransferase activity in the liver were observed. Activation of nitrogenous metabolism following glycerol injection is indicated by the increase of aminotransferase activity in organs, and concentration of blood urea. High concentration of creatinine in the rat serum can reflect malfiltration in kidneys.

[Antioxidant effects of L-arginine in the rat heart in experimental rhabdomyolysis].

The glycerol administration in a dose of 1 ml of 50% water solution/100 g b. w. was found to cause considerable accumulation of the total heme in the rat blood serum that is accompanied by an increase of TBA-reactive products and protein carbonyl derivates contents and by changes of protein level. Heme entering in the heart tissue is observed in the first hours after glycerol injection. The breaches of heart antioxidant-prooxidant balance are noted in twenty-four hours: TBA-reactive products and protein carbonyl derivates accumulation, heme oxygenase and catalase activation, superoxide dismutase activity lowering and reduction of glutathione content elevation. Pretreatment by L-arginine (0.5 h before glycerol administration) almost did not affect the blood serum changes caused by glycerol injection. However in the rat heart L-arginine administration prevents from TBA-reactive products and protein carbonyl derivates accumulation and the breaches of superoxide dismutase and catalase activities. Besides L-arginine causes the ealier heme oxygenase induction. Possible mechanisms of L-arginine protective action in the rat heart under experimental rhabdomyolysis are discussed.

[Effect of L-arginine on pro- and antioxidant status of the rat vessels and lungs in experimental rhabdomyolysis].

The glycerol administration was found to cause accumulation of the total heme in rat blood serum, vessels and lungs that are accompanied by increase of TBA-reactive products and protein carbonyl derivates contents. A decrease of superoxide dismutase activity and an increase of reduced glutathione in lung were observed. Heme entering the vessels and lungs is accompanied by elevation in heme oxygenase activity. Pretreatment by L-arginine (0.5 h before glycerol administration) didn't affect blood serum and vessels changes caused by glycerol injection. However, in lungs, L-arginine prevents TBA-reactive products and protein carbonyl derivates accumulation, the decrease ofsuperoxide dismutase activity and causes the ealier heme oxygenase induction. Prooxidant effects of heme in tissues studied and possible mechanisms of L-arginine protective action in lung under experimental rhabdomyolysis are discussed.

[Antioxidative status changes in golden syrian hamsters with experimental metabolic syndrome].

Some indices of the antioxidant status (content of the alpha-tocopherol, reduced glutathione and ascorbic acid, activity of the glutathione reductase and aryl-esterase) and lipid peroxidation processes in the liver, blood serum, and some blood serum lipoprotein fractions of the Golden Syrian hamsters of different sex and age status under high-caloric diet were investigated. It has been shown that the hypercaloric diet leads to a decreaseng of reduced glutathione content and increase of the level of lipid peroxidation products in the liver of experimental animals. The ascorbic acids content in male liver is decreased and in female liver is increased. In the blood serum under hypercaloric nutrition the accumulation of lipid peroxidation products and alpha-tocopherol content a decrease in ApoB-lipoproteins and HDL is observed. Simultaneously the ascorbic acid content is increased in the blood serum of all experimental animals. Activation of free-radical oxidation both in the liver, and blood serum is more significant in males compared with females. The data obtained allow to suppose that atherosclerotic complications of metabolic syndrome development may be connected to the lipoprotein oxidant status infringement.

[Heme oxygenase activity in the tissues of the vessels and heart of rats under co-administration of NO-synthase inhibitor and hemin chloride].

The administration of hemin chloride in a dose of 1.5 mg/100 g of the body weight was found to cause accumulation of the total heme and TBA-reactive products in the rat blood serum and vessels. Pretreatment by N(omega)-nitro-L-arginine (0.5 h before hemin chloride administration) did not affect the dynamics of the total heme and TBA-reacting products accumulation. The increase of heme oxygenase activity was observed in the vessels after hemin chloride administration. This effect was strengthened by N(omega)-nitro-L-arginine pretreatment. The changes of heme oxygenase activity and the total heme level in heart were not observed at any periods studied. The increase of the TBA-reactive products level in the heart after exogenous hemin injection was accompanied by an increase of nitrites content and blocked by pretreatment of NOS inhibitor. The N(omega)-nitro-L-arginine alone caused the accumulation of the total heme, TBA-reacting products and the increase of heme oxygenase activity in the vessels. The role of heme and NO in regulation of the heme oxygenase activity is discussed.

[Effect of heavy metals on activity of key enzymes of glyoxylate cycle and content of thiobarbituric acid reactive substances in the germinating soybean Glicine max L.seeds].

The influence of CoCl2 and CdCl2 on the activity of isocytrate lyase, malate synthase and NAD-malate dehydrogenase in the seed lobes and the composition of malondialdehyde products at early stages of germinating of soybean seeds: after first 24-hours, 72 hours and 96 hours are investigated. It is shown that when germinating in the medium containing no metal salts, isocytrate lyase activity is greatly increased during 96 h and malate synthase is increased after 72 h and is decreased after 96 h of germination period. CoCl2 activated isocytrate lyase activity after 72 hours and decreased malate synthase activity after 96 hours. The lengthening of the primary root under such conditions is noted. CdCl2 inhibited isocytrate lyase activity during first 24 hours and suppressed malate synthase activity after 96 hours. During this process the germ growth is suppressed. CoCl2 increased the composition of malondialdehyde products during each period of germination, and CdCl2 increased malondialdehyde content after 72 and 96 hours. The role of glyoxylate cycle enzymes in transforming fatty acids into carbohydrates and in forming the primary root under the process of germination of seed lobes of soybean is discussed.

[Effect of chronic social stress on lipid metabolism in golden Syrian hamsters].

The changes of total lipids, lipoproteins and their fractions, free fatty acids, triacylglyceroles, free and esterified cholesterol levels and parameters of its metabolism in the blood serum and liver, glucose-6-phosphate dehydrogenase and lysosomal lipase activity in the liver, and also post-heparin lipases activity in blood of hamsters with chronic social stress are investigated. Is has been shown, that in stressed animals the prevalence in early terms of chronic stress lipolysis above lipogenesis is observed. In later terms of chronic stress the lipogenesis activation is also observed which, alongside with active lipolysis, can cause hyperlipidemia in blood. The latter phenomenon is obviously more characteristic of males, while in females the main source of fatty acids in blood is probably lipolysis in the liver. Proatherogenic redistribution of lipoprotein fractions, which was observed at chronic stress, becomes complicated by changes of their transformations processes under blood lipases action, in particular, lipases disbalance: by increasing of hepatic lipase activity without lipoprotein lipase activity increase. The increase of CETP activity in HDL, which is observed at stress, can be accompanied by atherogenic LDLB accumulation in the blood plasma. The chronic social stress is proatherogenic owing to lipid and lipoprotein metabolism changes, which lead to the shift of balance during lipids transport and their use by tissues.

[V.N. Karazin Kharkov National University as the foundation of biochemistry in Ukraine].

The paper deals with the data on foundation and development of physiological chemistry (biochemistry) as independent science and education subject in the V. N. Karazin Kharkov National University before and after the organization of the Department of Physiological Chemistry. Studying the chemistry of natural compounds, their qualitative and quantitative content and transformations in living organisms both by foreign and home researchers made the basis for the appearance of physiological chemistry as static biochemistry. The improvement of the investigation methods and further discoveries caused the appearing of new branches--dynamic and functional biochemistry. The attention is paid to the fact that biochemistry arised at the Kharkov University as the education subject (A. I. Khodnev) and then developed as independent science due to efforts of A. Ya. Danilevskiy as well as biochemical school created by him. The Kazan' and Kharkov periods of scientific activity of A.Ya. Danilevskiy are described. The leading role of A. Ya. Danilevskiy in development of the home school of biochemistry is considered. Important role of A. V. Palladin in the foundation of Kharkov biochemists' school and organization of the Scientific-research Institute of Biochemistry in Kharkov is considered as well. It is stated that the Institute of Biochemistry after its arrival to Kiev and joining the Academy of Sciences became the center of Ukrainian biochemistry. The role of A. V. Nagorny and I. N. Bulankin in further development of biochemistry and foundation of a new scientific branch--age-related physiology and biochemistry--at the Kharkov University after its re-organization is discussed.

Affinity chromatography of GroEL chaperonin based on denatured proteins: role of electrostatic interactions in regulation of GroEL affinity for protein substrates.

The chaperonin GroEL of the heat shock protein family from Escherichia coli cells can bind various polypeptides lacking rigid tertiary structure and thus prevent their nonspecific association and provide for acquisition of native conformation. In the present work we studied the interaction of GroEL with six denatured proteins (alpha-lactalbumin, ribonuclease A, egg lysozyme in the presence of dithiothreitol, pepsin, beta-casein, and apocytochrome c) possessing negative or positive total charge at neutral pH values and different in hydrophobicity (affinity for a hydrophobic probe ANS). To prevent the influence of nonspecific association of non-native proteins on their interaction with GroEL and make easier the recording of the complexing, the proteins were covalently attached to BrCN-activated Sepharose. At low ionic strength (lower than 60 mM), tight binding of the negatively charged denatured proteins with GroEL (which is also negatively charged) needed relatively low concentrations (approximately 10 mM) of bivalent cations Mg2+ or Ca2+. At the high ionic strength (approximately 600 mM), a tight complex was produced also in the absence of bivalent cations. In contrast, positively charged denatured proteins tightly interacted with GroEL irrespectively of the presence of bivalent cations and ionic strength of the solution (from 20 to 600 mM). These features of GroEL interaction with positively and negatively charged denatured proteins were confirmed by polarized fluorescence (fluorescence anisotropy). The findings suggest that the affinity of GroEL for denatured proteins can be determined by the balance of hydrophobic and electrostatic interactions.

[Glucose-fatty acids cycle in cobalt chloride-induced oxidative stress in rats].

It was found that the glucose-fatty acids cycle functioned under the oxidative stress, caused by injection of cobalt chloride solution in albino rats. This cycle promoted the adaptation of metabolism and rehabilitated the homeostasis under extreme conditions. Its functioning was regulated by prolonged (during 2-24 hours) rise in activity of amino acids catabolism enzymes (e.g. tyrosine aminotransferase, arginase) and activation of glyconeogenesis after the mobilisation of liver glycogen. This contributed to increase in glucose and free fatty acids contents in blood. The latter is additionally provided by lipid mobilisation under stress. Tyrosine aminotransferase activation occurred both on the transcription level and by enabling of other mechanisms, which probably concerned the stabilisation of this enzyme. Preliminary injection of alpha-tocopherol in vivo significantly decreased the rise in tyrosine aminotransferase and arginase activities and the rate of erythrocyte hemolysis but did not disable them in full. This made evident that in regulation of the glucose-fatty acids cycle not only active metabolites of oxygen but also Co ions were directly enabled.

[Heme oxygenase induction in rat heart and vessels and peroxidative resistance of erythrocytes during hemolytic anemia development].

The hemolytic anemia development caused by phenylhydrazine injection (7 mg/100 g b.w.) was shown to be caused by the decreasing of both catalase activity and glutathione content in erythrocytes, and by the increasing of spontaneouse hemolysis level of these cells in blood stream. The increasing of heme oxygenase activity and TBA-active products in rat heart and vessels were revealed 24 hrs after phenylhydrazine injection. Possible mechanisms of heme oxygenase-1 induction under hypoxia as response to the hemolytic anemia development and it's role in defense of the cells from damage are discussed.

Free heme pool and activity of key enzyme of heme synthesis in the rat liver under action of agents affecting reduced glutathione level.

The decrease of GSH level in the rat liver was found to be accompanied by an increase of tryptophan 2,3-dioxygenase (TDO) heme saturation during first hours after HgCl2, phenylhydrazine (Ph) injection or rhabdomyolysis (the coefficient of correlation -0.978). The activity of the key enzyme of heme synthesis--5-aminolevulinate synthase (ALAS) was 2.5-fold increased in the first hours after Ph injection and rhabdomyolysis. Glutathione injection in vivo as well as CdCl2 caused the increase of GSH content and the inhibition of ALAS. The coefficient of correlation for GSH content and ALAS activity under the action of agents altering both these parameters (CdCl2, Ph, GSH injection and rhabdomyolysis) is 0.938. Taking into account the presence of heme regulatory motif with conserved cystein in many proteins, including ALAS and TDO (accession number in SwissProt database AAH61793 and P21643, respectively), the link between alterations of GSH content, ALAS activity and heme saturation of TDO in the rat liver could be proposed. The further experiments should be performed in order to elucidate the mechanisms of GSH level influence on free heme pool formation in the liver cells.

[Influence of cadmium chloride and hydrogen peroxide on the content of phosphoinositides in isolated hepatocytes of rats]. / Vliianie khlorida kadmiia i peroksida vodoroda na soderzhanie polifosfoinozitidov v isolirovannykh gepatotsitakh krys.

Influence of cadmium chloride and hydrogen peroxide on the processes of lipid peroxidation and contents of polyphosphoinositides in isolated hepatocytes of rats was studied. It is shown that incubation of the cells with cadmium chloride or hydrogen peroxide already in 15 min results in the reinforcement of lipid peroxidation and increase of contents of phosphatidylinositol and phosphatidylinositoldiphosphate. Variations of the basal level of the latter, as is well known, plays an important messenger role. In case of more long incubation (60 min) one can observe the significant accumulation of intermediate and final products of lipid peroxidation, while the content of phosphatidylinositol and phosphatidylinositoldiphosphate did not differ from the control, but the content of phosphatidylinositolphosphate was increased. Changes in the content of separate fraction of polyphosphoinositides in the presence of cadmium chloride and hydrogen peroxide were similar. Allowing for all that it is, possible to draw a conclusion that the mechanism of cadmium influence on the content of polyphosphoinositides is based on its ability to cause development of the oxidative stress.

[The effect of cadmium chloride and hydrogen peroxide on the lipid peroxidation and fractional composition of lipids in hepatocytes of rats]. / Vplyv khlorydu kadmiiu ta peroksydu vodniu na protsesy peroksydnoho okyslennia i fraktsiinyi sklad lipidiv u hepatotsytakh shchuriv.

The isolated hepatocytes were incubated in the medium, containing cadmium chloride or hydrogen peroxide. Influence of the latter on the intensity of lipid peroxidation and contents of some lipids fractions, as well as viability of hepatocytes in these conditions has been studied. It is shown that under such cultivation conditions the activation of lipid peroxidation in the hepatocytes takes place. Its activation in presence of cadmium chloride was one of the factors of the membranes damage. The changes in the content of some fractions of lipids were similar both under the incubations of the cells with cadmium chloride and hydrogen peroxide. This allows one to suppose that cadmium chloride causes changes in the lipid composition of membranes as a result of intensification of lipid peroxidation.

[Effect of cadmium chloride on polyphosphoinositides content in the rat liver and kidneys]. / Vliianie khlorida kadmiia na soderzhanie polifosfoinozitidov v pecheni i pochkakh krys.

The influence of cadmium chloride on the content of some fractions of polyphosphoinositides in the liver and kidneys of rats has been investigated in the work. We have reported that a single administration of sublethal dose of cadmium chloride leads to the long-term elevation of the content of diacylglycerol, which is responsible for the activation of protein kinase C. The increase of triphosphoinositides fraction content may be connected with activation of phosphoinositid-3-kinase and with accumulation of phosphatidylinositol-3,4-diphosphate and phosphatidilinositol-3,4,5-triphosphate, which are known as activators of some protein kinase C isoforms and also play an important role in the mitogen-activated protein kinase signaling pathway. The lipids fractions content changes were similar in the liver and kidneys, but had different time of response.

[Activity of key enzymes of heme metabolism and cytochrome P-450 content in the rat liver in experimental rhabdomyolysis and hemolytic anemia]. / Aktivnost' kliuchevykh fermentov metabolizma gema i soderzhanie tsitokhroma P-450 v pecheni krys pri éksperimental'nykh rabdomiolize i gemoliticheskoi anemii.

The 5-aminolevulinate synthase, heme oxygenase, tryptophan-2,3-dioxygenase activities, the content of total heme and cytochrome P-450 content in the rat liver and absorption spectrum of blood serum in Soret region under glycerol model of rhabdomiolisis and hemolytic anemia caused by single phenylhydrazine injection have been investigated. The glycerol injection caused a considerable accumulation of heme-containing products in the serum and the increase of the total heme content, holoenzyme, total activity and heme saturation of tryptophan-2,3-dioxygenase, as well as the increase of the 5-aminolevulinate synthase and heme oxygenase activities in the liver during the first hours of its action and the decrease of cytochrome P-450 content in 24 h. Administration of phenylhydrazine lead to the increasing of hemolysis products content in blood serum too, although it was less expressed. The phenylhydrazine injection caused the increase of activities of 5-aminolevulinate synthase, holoenzyme, total activity and heme saturation of tryptophan-2,3-dioxygenase, as well as decrease of cytochrome P-450 content in the rat liver in 2 h. The increase of the total heme content and heme oxygenase activity has been observed in 24 h. The effect of heme arrival from the blood stream, as well as a direct influence of glycerol and phenylhydrazine on the investigated parameters are discussed.

[Effect of ADP and GroES on interaction of molecular chaperonin GroEL with non-native lysozyme]. / Vliianie ADP i GroES na vzaimodeistvie molekuliarnogo shaperonina GroEL s nenativnym lizotsimom.

The interaction of the molecular chaperonin GroEL with fluorescein-labeled lysozyme in the presence of high concentrations of thiol reagent--dithiothreitol (DTT) has been studied. In case of high concentrations of DTT lysozyme loses the native conformation due to the disruption of the intramolecular disulfide bonds stabilizing its structure and effectively aggregates. It has been shown that in the presence of high concentrations of DTT and two-fold molar excess of GroEL the lysozyme tightly interacts with GroEL that essentially decreases the efficiency of its aggregation. The addition of ADP to the complex of GroEL with nonnative lysozyme noticeably decreases the interaction of the chaperonin with nonnative protein target resulting in some increase of the efficiency of its aggregation. However, the addition of the co-chaperonin GroES together with ADP (i.e. the formation of the complex of GroEL with GroES) leads to drastic weakness of the interaction of GroEL with nonnative lysozyme and the efficiency of its aggregation becomes comparable with that in the absence of GroEL.

[Activity of 5-aminolevulinate synthase in rat liver during degradation of cytochrome P-450 caused by administration of cadmium chloride]. / Aktivnost' 5-aminolevulinat-sintazy v pecheni krys v usloviiakh degradatsii tsitokhroma P-450 pri vvedenii khlorida kadmiia.

The 5-aminolevulinate synthase, tryptophan-2,3-dioxygenase activities and cytochrome P-450 content in the rat liver was studied in different terms after CdCl2 administration and after administration of metal salt against a background of 2-hours action of alpha-tocopherol. The lowering of activity of 5-aminolevulinate synthase in 2 h with the consequent increase of the enzyme activity in 6 h and 24 h was detected. The holoenzyme activity and heme saturation of tryptophan-2,3-dioxygenase increased 6 h after CdCl2 administration. The holoenzyme activity and the total activity of tryptophan-2,3-dioxygenase rised in 24 h. The level of cytochrome P-450 lowered. Preliminary administration of alpha-tocopherol prevented changes of studied parameters 24 h after CdCl2 administration. The relationship between decrease of cytochrome P-450 level and 5-aminolevulinate synthase activation are discussed.

[Activity of Ca(2+)-dependent neutral proteinases in rat organs under cobalt and mercury chloride injection]. / Aktivnost' Ca(2+)-zavisimykh neitral'nykh proteinaz v organakh krys pri vvedenii im khloridov kobal'ta i rtuti.

The activity of Ca(2+)-dependent neutral proteinases in rats under cobalt and mercury chloride injection was investigated. The calpains activity increase in the lungs, heart, liver and kidneys was revealed after 2 h cobalt chloride action. The mercury chloride gives a reliable increase of calcium-dependent neutral proteinases only in the kidneys.

[Metabolism of heme and hemeproteins and some indices of the antioxidant system in rat erythrocytes and tissues under anemia caused by phenylhydrazine]. / Metabolizm hemu ta hemoproteïniv i deiaki pokaznyky antioksydantnoï sistemy v erytrotsytakh i tkanynakh shchuriv pry fenilhidrazynovii anemiï.

The decrease of activity of several antioxidant enzymes in erythrocytes in the first hours after injection of phenylhydrazine to rats (7 mg per 100 g body weight) was found to be accompanied by accumulation of heme-containing compounds in rat serum and appearance of free heme in liver and decrease of cytochrome P450 content. Tissue-specific features of dynamics of activity of enzymes studied and reduced glutathione content were revealed, that might be caused by differences in total and free heme content in these organs. The role of key enzymes of heme biosynthesis and degradation in adaptation of metabolism under phenylhydrazine action is discussed.