[Vitamin D3 availability and functional activity of peripheral blood phagocytes in experimental type 1 diabetes].

The study was devoted to identifying the relation between vitamin D3 availability (assessed by the level of circulatory 25OHD3), content of vitamin D3 25-hydroxylase isozymes CYP27A1 and CYP2R1 in hepatic tissue and functional activity of peripheral blood phagocytes in mice with experimental type 1 diabetes. It has been shown that diabetes is accompanied by the development of vitamin D3-deficiency which is characterized by decreased 25OHD3 content in blood serum and determined by changes in tissue expression of the major isoforms of vitamin D3 25-hydroxylase. The level of hepatic CYP27A1 was revealed to be markedly reduced with a concurrent significant augmentation of CYP2R1. Cholecalciferol administration resulted in normalization of tissue levels of both isoforms of vitamin D3 25-hydroxylase and blood serum 25OHD3 content. Diabetes-associated vitamin D3 deficiency correlated with a decrease in phagocytic activity of granulocytes and monocytes, and their ability to produce antibacterial biooxidants such as reactive oxygen and nitrogen forms. Vitamin D3 efficacy to attenuate these abnormalities of immune function was established, indicating an important immunoregulatory role of cholecalciferol in the phagocytic mechanism of antigens elimination implemented by granulocytes and monocytes.

[The ROS-generating and antioxidant systems in the liver of rats treated with prednisolone and vitamin D3].

The mechanisms of glucocorticoid-induced disturbances of liver function is currently not fully clarified. Vitamin D3 was previously shown to play an important role in the regulation of impaired oxidative metabolism and detoxification function of the liver associated with the effects of hepatotoxic compounds. The study was undertaken to define the intensity of oxidative metabolism in the rat liver and survival of hepatocytes after prolonged prednisolone administration and to assess whether vitamin D3 is capable to counter glucocorticoid-induced changes. It has been shown that prednisolone (0.5 mg per animal for 30 days) leads to 1.6-fold increase in the percentage of necrotic cells among isolated hepatocytes as compared with the control. The glucocorticoid-induced impairment of hepatocellular function was accompanied by enhanced generation of reactive oxygen species (ROS), accumulation of TBA-active products and carbonylated proteins but reduced levels of free SH-groups of low molecular weight compounds. It was demonstrated a decrease in the activities of key enzymes of antioxidant system (SOD, catalase, glutathione peroxidase), whereas the activities of pro-oxidant enzymes NAD(P)H-quinone oxidoreductase and semicarbazide-sensitive amine oxidase were shown to be increased. Vitamin D3 (and to greater extent in combination with α-tocopherol) administration (100 IU) on the background of glucocorticoid therapy caused normalizing effects on the level of ROS formation, oxidative modification of biomolecules and activity of antioxidant enzymes resulting in better survival of hepatocytes. These data suggest a potential role of vitamin D3 in the regulation of oxidative metabolism alterations related to hepatotoxic action of glucocorticoids.

[Immunomodulatory effects of vitamin D3 and bisphosphonates in nutritional osteoporosis in rats].

The aim of this study was to determine the effectiveness of separate and combined administration of vitamin D3 and different forms of bisphosphonate (disodium salt of methylenbisphosphonic acid dihydrate and alendronate) on the function of immune cells in rats with nutritional osteoporosis. It was shown that D-hypovitaminosis leads to reduced 25OHD3, which is a biomarker for vitamin D3 and disturbances of metabolic processes in bone tissue that correlated with osteoporosis manifestation. Immunologic disorders related to nutritional osteoporosis were accompanied by the decrease in phagocytic activity of granulocytes and impaired ability to produce bactericidal oxidants. Inhibition of B-cell immunity also occurred in patholgy. Thus, the present study revealed more pronounced immunomodulatory effects of vitamin D3 on phagocytic immunity, whereas bisphosphonates were effective in improving the humoral immune protection.

[Role of PARP and protein poly-ADP-ribosylation process in regulation of cell functions].

This review focuses on the biological role of enzymes involved in posttranslational modification of proteins by their poly-ADP-ribosylation, a NAD-consuming process with an emerging key role in providing fundamental cell functions. To this end, detailed analysis of structural organization in relation to basic functions of the poly(ADP-ribose) polymerase-1 (PARP-1), the founding member of the PARP family, and other poly(ADP-ribose) polymerase isoforms is presented here. These include the current views on the role of PARP family enzymes and processes of poly-ADP-ribosylation of proteins in chromatin structure remodeling, DNA damage repair, regulation of gene expression, and integration of cellular signaling pathways. Considerable attention is paid to the involvement of PARP in cellular functions, particularly in cell division, intracellular transport of macromolcules, proteasomal protein degradation, immune response and caspase-independent necrotic pathways defined as necroptosis (programmed necrosis). In the light of the remarkable successes that have been reported for treating inflammatory disorders and cancer with different classes of PARPs inhibitors, we discuss the prospects of targeting PARPs with therapeutic purposes.

[Features of cholecalciferol hydroxylation in the liver of rats in conditions of D-hypervitaminosis and activity of alpha-tocopherol].

It is shown, that hepatocytes contain two (microsomal and mitochondrial) vitamin D3 25-hydroxylase enzymes, which differ as to their activity and function with maximal activity at different concentrations to substrate, namely at 15 microM and 100 microM of vitamin D3, accordingly. Activity of vitamin D3 25-hydroxylase enzymes of hepatocytes is regulated by cholecalciferol and alpha-tocopherol. The general and microsomal vitamin D3 25-hydroxylase enzymes activity of hepatocytes is lowered, but mitochondrial isoform is increased under D-hypervitaminosis conditions. Vitamin E increases microsomal vitamin D3 25-hydroxylase activity and decreases mitochondrial isoform activity of rats hepatocytes under D-hypervitaminosis conditions. It is established that D-hypervitaminosis is accompanied by expressed hypercalcemia and hyperphosphatemia, by decreased contents of mineral components in the bone tissue and high activity of alkaline phosphatase in the blood serum. The physiological doses of vitamin E under these conditions normalized the mineral metabolism, contents of calcium, phosphates and activity of alkaline phosphatase isoform in the blood serum.

[Role of vitamin E in regulation of cholecalciferol hydroxylation in hypovitaminosis D and hypervitaminosis D].

It is established, that dose-dependent influence of vitamin E on vitamin D3 metabolism, is conditioned by degree of cholecalciferol sufficiency. Under a condition of D-hypovitaminosis, contents of 25OHD3 in blood serum is 2-fold reduced and vitamin D3 25-hydroxylase enzymes activity increased in rat hepatocytes. Vitamin E (0.726-7.26 IU) significantly stimulated the effect of vitamin D3 (40 IU) in animals with D-hypovitaminosis and led to further increase of 25OHD3 content in the blood serum and activity of vitamin D3 25-hydroxylase enzymes in hepatocytes. In D-hypervitaminosis the contents of 25OHD3 in blood serum was more than 3-fold increased and vitamin D3 25-hydroxylase enzymes activity was inhibited. Vitamin E (0,726-7,26 IU) lowered the vitamin D toxicity, decreased contents of 25OHD3 in blood serum and activity of vitamin D3 25-hydroxylase enzymes in hepatocytes. High doses of vitamin E (36.3 IU) under these conditions demonstrated negative effect on vitamin D3 metabolism. The mechanism of vitaminE participation in the vitamin D3 metabolism under D-hypovitaminosis and D-hypervitaminosis may be its influence on the activity of different vitamin D3 25-hydroxylase systems of hepatocytes.

[Adaptogenic effect of the vitamin D3 containing supplement "videchol" on glucose-6-phosphate dehydrogenase activity in erythrone of irradiated rats].

Two fast migrating, major, multiple molecular forms (MMF) of glucose-6-phosphate dehydrogenase [EC:1.1.1.49]: G-6-PDH-1 and G-6-PDH-2, and two minor forms: G-6-PDH-3 and G-6-PDH-4 were revealed in the electrophoregrams of both erythrocytes haemolisates as well in the homogenates of bone marrow cellular lines of rats at control conditions. Daily 1 cGy irradiation of rats up to a cumulative dose of 20 cGy led to a drop of G-6-PDH total activity and it caused a redistribution of the MMF of the enzyme in bone marrow cellular populations. However, G-6-PDH activity in erythrocytes exceeded the control means in all the experimental terms. The calculation of the local redistribution coefficient (l(G-6-FDH-i)) showed that these changes are mainly determined by the increase of the activity of the isoform G-6-PDH-3. Vitamin D3 administration to rats generated a correction of G-6-PDH activity in all studied cellular populations. Meanwhile, the MMF profiles were characterized by multidirectional rearrangements in the bone marrow erythroid and granulocyte-monocyte cells and in erythrocytes. The specificity of changes in the distribution of the MMF of G-6-PDH in the three studied cellular populations depends on the particularities of their energetic metabolism at irradiation conditions and on the modifying action of the natural adaptogen 1,25-dihydroxicholecalciferol.

[Effect of nicotinamide on the reactions of peroxide oxidation of biomolecules in the rat brain and erythrocytes in 1,2-dichloroethane toxic injury].

It was established that acute poisoning of rats by 1,2-dichloroethane induced considerable changes in lipid peroxidation indices, glutathione content and activity of antioxidant enzymes--superoxidase, catalase, glutathione peroxidase in the brain tissue, erythrocytes and blood plasma. It was shown that nicotinamide in the dose of 200 mg/kg prevented considerable degree of the intoxication caused by 1,2-dichloroethane as well as activation of lipid peroxidation and inhibition of antioxidant defens enzyme activities in tissue of experimental animals.

[Na+, K+-ATPase activity and serotonin transport in the rat brain synaptosomes fractions after acute 1,2-dichloroethane intoxication and nicotinamide administration]. / Na+, K+-ATR-azna aktyvnist' ta transportuvannia serotoninu u fraktsiï synaptosom holovnoho mozku shchuriv za hostroï intoksykatsiï 1,2-dykhloretanom ta vvedennia nikotynamidu.

Alterations of Na+,K+-ATPase activity and serotoninergic system functioning were investigated in brain synaptosomes fractions of rats under experimental acute 1,2-dichloroethane (DChE) intoxication. It was shown that Na+,K+-ATPase activity was markedly increased (by 41,8%) in a period of 24 h after DChE intoxication and decreased (by 27%) after 48 h intoxication. The level of [2-14C]-serotonin uptake by synaptosomes was progressively diminished after 24 and 48 h after DChE injection whereas the activity of monoamine uptake proved to be unchanged. Nicotinamide (200 mg/kg of body weight) was administered to rats subjected to DChE 1, 24 and 36 h after poisoning. The treatment of rats with nicotinamide resulted in some normalization of brain synaptosomal Na+, K+-ATPase activity and serotonin uptake controlled at 48 h after DChE intoxication.

[Role of phosphatidylinositol 3-kinase in platelet aggregation in type 1 diabetes mellitus]. / Rol' fosfatydylinozytol-3-kinazy v rehuliatsiï ahrehatsiinoï zdatnosti trombotsytiv za tsukrovoho diabetu 1-ho typu.

It was found that incubation of platelet rich plasma with wortmannin, an irreversible selective inhibitor of phosphoinositide 3-kinase (PI3K), leads to sharp drop in platelet aggregation ability in healthy donors, whereas in type 1 diabetes mellitus patients this effect was less manifested or not quite determined. Translocation dynamics of PI3K regulatory subunit into cytoskeleton fraction under induction of platelet aggregation by various ADP concentrations and after wortmannin treatment was studied. Reciprocal interaction of endothelial constitutive NO synthase with PI3K in mechanisms of platelet functional state regulation under studied pathological and normal conditions have been analyzed.

[Catalase activity as affected by carnosine and sodium nitrite in vitro]. / Aktyvnist' katalazy za diï nitrytu natriiu ta karnozynu in vitro.

It was shown in vitro that the activity of catalase Penicillium vitale decreases when treated by sodium nitrite to the greater extent than when affected by dipeptide carnosine (beta-alanyl-L-histidine). At alternating introduction of carnosine and NaNO2 that component affected the catalase activity which was the first to be introduced. The entering of the next metabolite into the medium did not change the enzyme activity. It is shown that carnosine binds with a molecule of catalase. Carnosine may be considered one of natural regulators of catalase activity.

[Activity of antioxidant enzymes in enterocytes of the small intestine and blood cells after ionizing irradiation and administration of vitamin E in rats]. / Aktyvnist' fermentiv antyoksydantnoho zakhystu v enterotsytakh tonkoho kyshechnyka ta klitynakh krovi za ionizuiuchoho oprominennia ta vvedennia shchuram vitaminu E.

The long-term influence of low X-ray irradiation increases lipid peroxidation (LP) in radiosensitive (bone marrow, enterocytes of small intenstine) and in relatively radioresistant blood cells (erythrocytes). The activation of antioxidant system enzymes in observed cells does not decrease LP intensity. We concluded that additional administration of alpha-tocopherol provided the decrease of the first and end products of LP in the observed tissues mostly in the beginning of the experiment. Antioxidant effect of the preparation is more significant in cells with high proliferative activity but normal activity of enzymes was not determined.

[Oxidative stress correction by nicotinamide and nicotynol-GABA in diabetic neuropathy]. / Korektsiia nikotynamidom ta nikotynoïl-HAMK oksydatyvnoho stresu za diabetychnoï neiropatiï.

Concentration of lipid peroxidation products and antioxidant enzyme activities in rat brain and erythrocytes and the effects of nicotinamide and nicotinoyl-GABA administration on these parameters were estimated on 21st day of streptozotocin-induced diabetes. It was demonstrated more then two-fold diabetes-induced accumulation of conjugated dienes and malondialdehyde in tissues studied. Superoxide dismutase and glutathione reductase activities of both brain homogenate and erythrocytes as well as catalase and glutathione peroxidase activities of brain homogenate were shown to decrease significantly in diabetic rats, meanwhile, catalase activity of erythrocytes was increased and glutathione peroxidase unchanged. So the correlation between changes in enzymatic antioxidant system in brain and erythocytes failed to be found. Alterations observed were virtually prevented by the course of nicotinamide and nicotinoyl-GABA treatment. The results suggested that the suppression of antioxidant system could be primary biochemical disturbance in diabetic neuropathy progression. It was shown that the antioxidant efficacy of nicotinoyl-GABA is lower than that of nicotinamide. It was suggested that the mechanism of antioxidant action of nicotinamide and its structural analogue consists of both scavenging of lipid peroxides and NAD biosynthesis that leads to activation and normalization of altered energy and lipid metabolism.

[Scientific-Pedagogic School of Biological and Medical Chemistry of the O. O. Bogomolets National Medical University (on the 160th year of its founding)]. / Naukovo-pedahohichna shkola biolohichnoï ta medychnoï khimiï Natsional'noho medychnoho universytetu im. O. O. Bohomol'tsia (do 160-richchia zasnuvannia).

In this work the most important stages of the scientific-pedagogic school of biologic and medical chemistry formation in Bogomolets National Medical University starting from the period of foundation as early as in 1863 till nowadays the Chair of Medical Chemistry and Physics as a part of Medical Faculty of Saint Volodymyr Emperor University in the city of Kyiv have been estimated and generalized. The especial attention is attracted to the fact, that it was Kyiv University where firstly the Chair of Biochemistry was created in order of stuyding the regularities of biochemical processes running in the human organism and metabolism disturbances inducing the pathologic processes at some diseases. The scientific and scientific-pedagogical trends of the chair work in different periods of its development are presented, simltneously the leading role of famous Ukrainian scientists--biochemicians in foundation and development of biologic and medical chemistry scientific school in the University are emphasized. Nowadays the Chair is the educational and scientific center supporting and developing the best traditions on training the specialists of different qualification levels: physicians Masters of Science, Philosophy Doctors and Doctors of Science in Medicine and Biology. The Chair is considered to be a basic one among the Ukraine higher medic and pharmaceutic educational institutions having the III-IV accreditation rate on the problems of teaching-organizational, educational-methodical and scientific work. On the Chair base there is functioning the Scientific Problem-Solving Commission of Ministry of health Protections of Academy of Medical Sciences of Ukraine "Biological and medical Chemistry" (the chairman is the Corresponding-Member of Academy of Medical Sciences of Ukraine, Prof. Yu.I. Gubsky. The Chair personnel compiled and issued the contemporary manuals in Ukraine language on Biologic and Bioorganic Chemistry.

[Diabetic neuropathies. Metabolism of sorbitol in sciatic nerve tissue in streptozotocin diabetes]. / Diabetychna neiropatiia. Metabolizm sorbitolu u tkanyni sidnychnoho nerva za streptozototsynovoho diabetu.

The increase of sorbitol and fructose levels caused by aldose reductase activation and sorbitol dehydrogenase inhibition were observed in sciatic nerve of streptozotocin-diabetic rats. Elevated polyol pathway activity has been implicated in the development of diabetic complications such as neuropathy. The regulation of polyol pathway enzymes is based on the changes of redox state of free nicotinamide nucleotides. The decrease of the NADP+/NADPH ratio in cytosolic compartment of sciatic nerve cells activated aldose reductase and the decrease of the NAD+/NADH ratio inhibited sorbitol dehydrogenase. Nicotinamide as a precursor of NAD+ biosynthesis increased the free NADP+/NADPH and NAD+/NADH ratios and inhibited the activity of polyol pathway. The sorbitol level decreased in sciatic nerve of nicotinamide-treated streptozotocin-diabetic rats as compared to non-treated ones. Thus, the data provide evidence for important role of nicotinamide, as an antidiabetic drug, in prevention or correction of diabetic neuropathy.

[Features of fibronectin-dependent activation of ribosomal protein S6 kinase (S6K1 and S6K2)]. / Osoblyvosti fibronektynzalezhnoï aktivatsiï kinaz rybosomnoho bilka S6 (S6K1 i S6K2).

Integrin family of adhesion receptors play an important role in organizing the actin cytoskeleton and in signal transduction from the extracellular matrix. The previous studies have shown that exposure of fibroblast cells to extracellular matrix proteins activates ribosomal S6 kinase 1 (S6K1) pathway in a ligand dependent manner. Recently, a new, highly homologous ribosomal S6 kinase, termed S6K2, was identified. It has 70% amino acid identity in the overall sequence with S6K1, and the potential phosphorylation sites of S6K1 are conserved in S6K2. However, the N- and C-terminal domains of S6K2 are quite different from those of S6K1. In this study we have examined dynamics of fibronectin-induced activation of these two kinases, transiently expressed in human HEK 293 cells. Differences between profiles of activation of S6K1 and S6K2 were observed in the early period of fibronectin stimulation. Fibronectin-induced changes in S6K2 activity were closely correlated with phosphorylation at Ser423, which is homologues to Ser 434 of S6K1. Although we didn't observe considerable changes in phosphorylation of S6K1 at Ser434, suggesting potential differences in the regulation of these homologous kinases upon fibronectin stimulation.

[Features of the inhibition of catalase activity by 3-amino-1,2,4-triazole in erythrocytes and liver of rats with streptozotocin diabetes]. / Osoblyvosti hal'muvannia aktyvnosti katalazy 3-amino-1,2,4-triazolom v erytrotsytakh i pechinsti shchuriv zi streptozototsynovym diabetom.

Lipid peroxidation was intensified by streptozotocin induced diabetes mellitus in erythrocytes and liver. Activity of antioxidant enzyme superoxide-dismutase was decreased, activity of catalase was increased. Concentration of lipid peroxidation products was decreased after nicotinamide injections. It was investigated liver- and erythrocyte catalase inhibition in the presence of 3-amino-1,2,4-triazole. Effective inhibitor concentration for liver catalase by streptozotocin induced diabetes mellitus was 10 mM, by control-20 mM. Ascorbic acid induced catalase inhibition in the erythrocytes by diabetes mellitus increased by ascorbic acid concentration from 25 to 150 mM. [DHAA]/[AA]-ratio increased from 0.26 by control to 1.6 by diabetes mellitus and decreased to 0.44 after nicotinamide injections.

[State of antioxidant and oxygen transport system of the blood in the process of adaptation of the body to hypoxic hypoxia]. / Stan antyoksydantnoï ta kysnevo-transportnoï system krovi v protsesi adaptatsiï orhanizmu do hipoksychnoï gipoksiï.

This article shows that in oxygen transported and antioxidative systems considerable changes take place during dynamic of adaptation to oxygen deficiency of inhaled air (every day 4 hours stay of animals in barochamber under pO2 7.45 kPa during 12 days). Single influence of hypoxic factor induced in erythrocytes peripheral blood increasing of haemoglobin affinity to oxygen, decreasing SOD and catalase activity on the background of increasing of malon aldehyde level. On 12-th day of adaptation of animals contents of TBA-active products in erythrocytes is saved on high level, activity of antioxidative enzymes is approached to original meanings, haemoglobin affinity to oxygen is decreased in comparison with control. Detected increasing content of creatine in erythrocytes during dynamic of rat adaptation to hypoxic hypoxia witnessed about erythropoiesis activation and quickened exit out of marrow young forms of erythrocytes.

[Physico-chemical characteristics and functional properties of membrane-bound hemoglobin]. / Fizyko-khimichna kharakterystyka i funktsional'ni vlastyvosti membranozv'iazanoho hemohlobinu.

It was shown that fetal form of haemoglobin (Hb) is mainly connected with membrane of peripheral red blood corpuscles. Membrane-bound haemoglobin has relatively high affinity to O2, low Hill's coefficient, much higher peroxidase activity in comparison with cytosolic Hb.

[The effect of nicotinamide on the enzymatic activity of the antioxidant defense in experimental diabetes]. / Vplyv nikotynamidu na aktyvnist' fermentiv antyoksydantnoho zakhystu pry eksperymental'nomu diabeti.

Experiments on rats with experimental streptosotocin-induced diabetes have shown intensification of the lipid peroxidation processes and reduction of activity of antioxidant defensive enzymes. The content of G-SH and glutathione peroxidase activity has decreased in comparison with the normal rate by 69% and 28%, respectively. Glutathione reductase activity has risen by 20%. Activity of the antioxidant enzymes (superoxide dismutase, catalase) has reduced and the amount of the final product of lipid peroxidation, MDA has increased. Injection of nicotinamide to diabetic rats (200 mg/1 kg of weight) for 14 days normalized activity of the antioxidant enzyme system and the content of the lipid peroxidation products.