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Showing papers in "Ukraïns'kyĭ biokhimichnyĭ zhurnal in 2010"


Journal Article
TL;DR: Purine-purine mispairs of DNA leading to pyrimidine-purines "transversions"-type point mutations were revealed and characterized at the MP2/6-311++G(2df,pd)//B3LYP(d,p) level of theory in vacuum approach adequately modeling hydrophobic environment of the active centre of high-fidelity replicative DNA-polymerases.
Abstract: Purine-purine mispairs of DNA (thus involving template base in anti-conformation along the glycosidic bond and base of the incoming nucleotide - in syn-conformation) leading to pyrimidine-purine "transversions"-type point mutations were revealed and characterized at the MP2/6-311++G(2df,pd)//B3LYP/6-311++G(d,p) level of theory in vacuum approach adequately modeling hydrophobic environment of the active centre of high-fidelity replicative DNA-polymerases.

24 citations


Journal Article
TL;DR: A novel physico-chemical mechanism of the Watson-Crick DNA base pair Gua.Cyt tautomers of bases are revealed and realized in a pathway of single proton transfer through two mutual isoenergetic transition states with Gibbs free energy of activation.
Abstract: A novel physico-chemical mechanism of the Watson-Crick DNA base pair Gua.Cyt tautomerization Gua.Cyt* Gua.Cyt Gua*.Cyt (mutagenic tautomers of bases are marked by asterisks) have been revealed and realized in a pathway of single proton transfer through two mutual isoenergetic transition states with Gibbs free energy of activation 30.4 and 30.6 kcal/mol and they are ion pairs stabilized by three (N2H...N3, N1H...N4- and O6+H...N4-) and five (N2H...O2, N1H...O2, N1H...N3, O6+H...N4- and 06+H...N4-) H-bonds accordingly. Stable base pairs Gua-Cyt* and Gua*.Cyt which dissociate comparably easy into monomers have acceptable relative Gibbs energies--12.9 and 14.3 kcal/mol--for the explanation of the nature of the spontaneous transitions of DNA replication. Results are obtained at the MP2/6-311++G(2df,pd)//B3LYP/6-31 1++G(d,p) level of theory in vacuum approach.

24 citations


Journal Article
TL;DR: The viability of normal (Wistar rat thymocytes) and transformed T cells after UV/Vis irradiation in the presence of pristine C60 fullerene was studied and may be useful for development of photosensitizers for photodynamic therapy with selective action on leukemia cells.
Abstract: The viability of normal (Wistar rat thymocytes) and transformed (human leukemia Jurkat cells) T cells after UV/Vis irradiation in the presence of pristine C60 fullerene was studied. The data obtained have shown that C60 fullerene exhibits cytotoxic effect against transformed T lymphocytes when combined with UV/Vis irradiation using mercury-vapor lamp (320-600 nm). C60 fullerene photocytotoxicity was not detected in thymocytes. C60-dependent photoinduced apoptosis of Jurkat cells was confirmed by DNA fragmentation and caspase-3 activation. No substantial increase of caspase-3 activation was observed in thymocytes treated with C60 fullerene plus irradiation, while antileukemic agent cytosine arabinoside was shown to induce caspase-3 activation both in Jurkat cells and thymocytes. The data obtained may be useful for development of photosensitizers for photodynamic therapy with selective action on leukemia cells.

11 citations


Journal Article
TL;DR: The obtained data indicate that the active sites on SPM responsible for thiamine binding and ThTPase activity have different sensitivity toThiamine antagonists, which allow us to suppose that different active protein sites are responsible for the specific binding and for th Liamine phosphates hydrolysis by TBP of synaptic membranes.
Abstract: The current work is aimed at understanding the structure and functionality of thiamine binding protein (TBP) in neural cells plasma membranes. The influence of thiamine triphosphate on thiamine binding by TBP in synaptic plasma membranes (SPM) isolated from the rat brain was investigated. It was shown that thiamine triphosphate inhibits thiamine binding activity of SPM in concurrent manner (K(i) = 1.0 +/- 0.3 microM). At the same time thiamine had no effect on thiamine triphosphatase (ThTPase) activity at the concentration range 0.5-20 microM. Otherwise, ThTPase activation with the maximum at the concentration about 2.5 microM was observed. Further, the influence of classic thiamine antagonists (amprolium, oxythiamine and pyrithiamine) on both biological activities of TBP in SPM was studied. The IC50 value for inhibition of thiamine binding on SPM by amprolium comprised 50 +/- 4.0 microM. Still, this antagonist had no effect on ThTPase activity. For the oxythiamine inhibition of both TBP activities was detected. The values of IC50 were 125 +/- 28 and 1000 +/- 95 microM for thiamine binding and ThTPase activity, respectively. The values of IC50 for thiamine binding and ThTPase activity inhibition differed by more than one order of magnitude and comprised 2.2 +/- 0.2 and 43 +/- 9 microM, respectively. The obtained data indicate that the active sites on SPM responsible for thiamine binding and ThTPase activity have different sensitivity to thiamine antagonists. Our results allow us to suppose that different active protein sites are responsible for the specific binding and for thiamine phosphates hydrolysis by TBP of synaptic membranes.

8 citations


Journal Article
TL;DR: In this paper, it was shown that in addition to traditional incorporational errors caused by facilitated (compared with the canonical DNA bases cytosine (Cyt)) tautomerization of 6-(2-deoxy-beta-D-ribofuranosyl)-3,4-dihydro-6H,8H-pyrimido[4,5-c][1,2]oxazin-7-one (DCyt), this mutagen causes the replication errors, increasing one million times the population of mispair Gua.
Abstract: Using the simplest molecular models at the MP2/6-311++G(2df,pd)//B3LYP/6-311++G(d,p) level of the theory it has been shown for the first time that in addition to traditional incorporational errors caused by facilitated (compared with the canonical DNA bases cytosine (Cyt)) tautomerization of 6-(2-deoxy-beta-D-ribofuranosyl)-3,4-dihydro-6H,8H-pyrimido[4,5-c][1,2]oxazin-7-one (DCyt), this mutagen causes the replication errors, increasing one million times the population of mispair Gua.DCyt* (asterisk marked mutagenic tautomer) as compared with mispair Gua.Cyt*. It is also proved that DCyt in addition to traditional incorporational errors also induces similar errors by an additional mechanism - due to a facilitated tautomerization of the wobble base pair Ade.DCyt (compared to the same pair Ade.Cyt) to a mispair Ade.DCyt* which is quasirisomorphic Watson-Crick base pair. Moreover, the obtained results allowed interpreting non-inconsistently the existing experimental NMR data.

7 citations


Journal Article
TL;DR: The alpha-domain is responsible for the increased release of metals from injured MTs in frogs, whereas extremely high oxidizability of beta-domain makes its participation in the exchange of metals elusive and provokes the aggregation of MTs.
Abstract: The metal-buffering and stress proteins metallothioneins (MTs) of frog are characterised by unusually high content of copper as for vertebrate animals and instability that was shown in our previous studies. They easily lost copper and especially zinc under unfavourable conditions. The aim of this study was to examine the reactivity of SH groups in the MTs from the liver of frog Rana ridibunda after the effect of Cu2+ (0.01 mg/l) and Zn2+ (0.1 mg/l) ions on the organism during 14 days. The alpha- and beta-domains of MTs with molecular weights of about 4 kDa were separated by the size-exclusion chromatography on Sephadex G-50. Unlike higher vertebrates, frogs demonstrated higher reactivity of alpha-domain than beta-domain with the Ellman's reagent (DTNB). The signs of partial oxidations in beta-domain included the creation of by-products with molecular weight about 12 kDa, low reactivity of SH-groups, and typical of -S-S-bonds peculiarities of UV-spectra. The effect of both metal ions on frog provoked the elevation of SH-groups reactivity in a-domain with the appearance of by-product with molecular weight of 16 kDa and its reduction in beta-domain. The incubation of MTs of control animals with 0.5 and 5.0 mM of H2O2 did not affect its chromatographic characteristics. In the frogs loaded by Cu2+ and Zn2+ the effect of 5.0 mM H2O2 on MTs provoked the release of 4 kDa product. So the alpha-domain is responsible for the increased release of metals from injured MTs in frogs, whereas extremely high oxidizability of beta-domain makes its participation in the exchange of metals elusive and provokes the aggregation of MTs.

6 citations


Journal Article
TL;DR: It was shown that oligomycin significantly increases mitochondria granularity, but Mg2+ and RuR have no influence on this parameter.
Abstract: The influence of modulators of calcium exchange in mitochondria--oligomycin, Mg2+ and ruthenium red (RuR)--on the myometrium mitochondria size and granularity was studied in the work. The study of the mitochondria size was carried out using the photon correlation spectroscopy. It was shown that the average hydrodynamic diameter was 655 +/- 14 nm (n = 5; control). The addition of oligomycin (1 microg/ml)--the inhibitor of ATP-synthase F0-component, increases the mitochondria average hydrodynamic diameter to 913 +/- 75 nm (n = 5), that is by 39% more than the control. In the presence of RuR (10 microM) (Ca2(+)-uniporter inhibitor) and Mg2+ (7 mM) the mitochondria average hydrodynamic diameter increases to 788 +/- 28 and 788 +/- 38 nm (n = 5) respectively, that is by 17% more than the control. Using flow cytometry it was shown, that oligomycin (1 microg/ml) causes the increase of side scattering of the mitochondria. Addition of RuR (10 microM) and Mg2+ (7 mM) does not lead to significant changes in side scattering of the mitochondria. So it was shown that oligomycin significantly increases mitochondria granularity, but Mg2+ and RuR have no influence on this parameter

5 citations


Journal Article
TL;DR: In this article, a conformational analysis of the DNA structural unit was carried out with the applied quantum mechanics methods at the MP2/6-311++G(d,p) // B3LYP/6 -31G (d, p) theory level and 660 conformations with relative Gibbs energies from 0 to 11.1 kcal/mole have been found.
Abstract: The conformational analysis of the DNA structural unit--the nucleotide with thymine base and electroneutral phosphate group at 5'-position-has been carried out with the applied quantum mechanics methods at the MP2/6-311++G(d,p) // B3LYP/6-31G(d,p) theory level. As many as 660 conformations with relative Gibbs energies under standard conditions from 0 to 11.1 kcal/mole have been found. Among them, six conformations are similar to the structure of the nucleotide of AI-DNA, one--to AII- and seven--to the DNA in BI-form. The lowest Gibbs energy among the DNA-like conformations (deltaG = 2.7 kcal/mole) belongs to BI-DNA-like structure. It is shown that the glycoside chemical bond is the most labile one. The role of intramolecular CH...O hydrogen bonds in formation of the 5'-thymidilic acid molecule structure is demonstrated.

5 citations


Journal Article
TL;DR: Purified human arginase I preparations homogeneous in SDS-PAAG test were obtained by the affinity chromatography on the synthesized sorbent L-arginine-macroporous glass by studying the influence of some bivalent metal ions and other additives on enzymatic activity for stabilization of the enzyme and optimization of its storage conditions.
Abstract: Purified human arginase I preparations homogeneous in SDS-PAAG test were obtained by the affinity chromatography on the synthesized sorbent L-arginine-macroporous glass. Some physico-chemical characteristics of the isolated arginase preparation have been estimated: thermo- and pH-stability, temperature- and pH-optima of the enzyme. The influence of some bivalent metal ions and other additives on enzymatic activity for stabilization of the enzyme and optimization of its storage conditions was studied.

5 citations


Journal Article
TL;DR: It was shown that EGFP-SbB could interact with cell surface of both toxin-sensitive monkey cells (Vero cell line) and toxin-resistant mouse cells (3T3 cell line), and was able to interact with receptors of both cell lines and to internalize into these cells.
Abstract: The recombinant fluorescent derivative of diphtheria toxin (EGFP-SbB) obtained by the replacement of toxin A subunit by enhanced green fluorescent protein (EGFP) has been used for visualization of the interaction of diphtheria toxin (DT) with sensitive and insensitive cells. It was shown that EGFP-SbB could interact with cell surface of both toxin-sensitive monkey cells (Vero cell line) and toxin-resistant mouse cells (3T3 cell line). The affinity of this protein for receptors of Vero cells was three times higher as compared with 3T3 cells. It was demonstrated that fluorescent derivate was able to interact with receptors of both cell lines and to internalize into these cells. Internalization of EGFP-SbB into the cells was inhibited by endocytosis inhibitor phenyl arsine oxide. We suppose that diverse sensitivity to DT of monkey and mouse cells can be explained not only by differences in their receptor affinity for DT but also by the processes that occur after internalization of the toxin into the cells.

5 citations


Journal Article
TL;DR: Glycosylated flavonoids: rutin, naringin, baikalin and methylhesperidin were shown to inhibit furin at pH 7.2 reversibly and competitively with Ki- 80-200 microM, indicating a new class of small non-peptide inhibitors of furin.
Abstract: Furin, a human subtilisin-related proprotein convertase, is the most important pharmaceutical target because it plays a vital role in development of numerous disease processes. To identify a new class of small non-peptide inhibitors of furin we performed a study of several flavonoids and some natural products. Glycosylated flavonoids: rutin, naringin, baikalin and methylhesperidin were shown to inhibit furin at pH 7.2 reversibly and competitively with Ki- 80-200 microM. The Ki values were derived from Dixon and/or Eadie-Hofstee plots using fluorogenic substrate Boc-Arg-Val-Arg-Arg-AMC. Although studied flavonoids display only a temperate furin inhibition, they may serve as a great potential for the future development of more potent non-peptide inhibitors against furin.

Journal Article
TL;DR: The analysis of 1H NMR spectra revealed the overall balance in favor of apoptosis, namely the increase in the content of NMR-visible mobile lipid domains and the decreased intensity of choline-containing metabolites.
Abstract: This work was partly supported by the grant from the research program ‘‘Fundamentals of Genomics and Proteomics’’ No. 107U002244 of National Academy of Sciences of Ukraine. We would like to thank Dr. N. Khranovskaya for her help with flow cytometry.

Journal Article
TL;DR: In vitro tests revealed that derivatives of 2-thioxo-thiazolidin-4-one exhibited inhibitory activity against ASK1, and binding mode for inhibitors of this class with ASK 1 ATP-binding site was proposed, which can be used for further optimization and developing more potent and selective inhibitors of ASK2.
Abstract: Protein kinase ASK1 (Apoptosis signal-regulating kinase 1) plays a key role in cell differentiation, aging and apoptosis. High activity of the kinase is associated with several pathologies. The ASK1 inhibitors might be therapeutic for patients with neurodegenerative, cardiovascular diseases and fibrous histiocytoma. In this work the identification of ASK1 inhibitors was performed by the methods of computer modeling and biochemical testing in vitro. The virtual screening experiments were carried out targeting the ATP binding site of ASK1 by browsing the database which contained 164 840 compounds of diverse chemical classes. The best-scored 300 ligands have been taken for the kinase assay analysis. In vitro tests revealed that derivatives of 2-thioxo-thiazolidin-4-one exhibited inhibitory activity against ASK1. The most active compound was 5-bromo-3-(4-oxo-2-thioxo-thiazolidin-5-ylidene)-1,3-dihydro-indol-2-one (IC50 = 2 microM). Binding mode for inhibitors of this class with ASK1 ATP-binding site was proposed. Our results can be used for further optimization and developing more potent and selective inhibitors of ASK1.

Journal Article
TL;DR: It was established that PA changes essentially the 5-LO thermostability, namely activation energy Ea of enzyme denaturation increases several times in this phospholipid presence, which can evidence that PA interaction with 5- LO leads to conformational changes of enzyme probably due to hydrophobic interactions.
Abstract: The investigation aim was to establish the structural-functional relations of individual lipid metabolism components: enzymes--lipoxygenases and membrane phospholipids. Influence of phosphatidic acid (PA)--allosteric activator of potato tuber 5-lipoxygenase (5-LO)--on thermoinactivation thermodynamic parameters of enzyme was studied. It was established that PA changes essentially the 5-LO thermostability, namely activation energy Ea of enzyme denaturation increases several times in this phospholipid presence. Such changes can evidence that PA interaction with 5-LO leads to conformational changes of enzyme probably due to hydrophobic interactions. Obtained results give a possibility to interpret the action mechanism of PA as 5-LO allosteric activator, which can displace the substrate molecules in binding sites and increase the level of formation of specific products of linoleic acid oxygenation by 5-LO.

Journal Article
TL;DR: It is concluded that Src plays a key role in the mechanisms of increasing the colonic VP during experimental UC and protein-protein interaction between beta-arrestine2 and VE-cadherine was enhanced, that might be a reason of colonic endothelium barrier disruption.
Abstract: We have shown the increase of Src(Tyr416) phosphorylation in rat colonic mucosa at early stages of 6% iodoacetamide-induced ulcerative colitis (UC), while the level of Src protein expression was not changed. Pretreatment of rats with Src inhibitor PP1 (0.2 mg/100 g, subcutaneously) decreased the colonic vascular permeability (VP) (P < or = 0.001) and pSrc(Tyr416) level during iodoacetamide-UC. Iodoacetamide-induced autophosphorylation and upregulation of VEGFR-2 was associated with Src activation in colonic mucosa of rats. Sequentially, protein-protein interaction between beta-arrestine2 and VE-cadherine was enhanced, that might be a reason of colonic endothelium barrier disruption. We concluded that Src plays a key role in the mechanisms of increasing the colonic VP during experimental UC.

Journal Article
TL;DR: It was shown that fibronectin-1 mRNA expression in lymphocytes is increased in patients with erythremia as compared to healthy donors and is accompanied by a decrease of fibronECTin plasma level.
Abstract: Expression of fibronectin-1 mRNA in lymphocytes in erythremia patients and healthy donors as well as fibronectin-1 concentration in plasma and its heparin-binding activity have been studied. Moreover, we also investigated the expression of fibronectin protein in lymphocytes and cell surface in erythremia disease as compared to healthy donors. It was shown that fibronectin-1 mRNA expression in lymphocytes is increased in patients with erythremia as compared to healthy donors. The decrease of plasma fibronectin concentration and its heparin-binding activity as well as the increase of lymphocyte content with surface-associated and intracellular fibronectin were revealed in erythremia disease in comparison with healthy donors. Positive correlation between plasma fibronectin level and its heparin-binding activity and negative correlation between plasma fibronectin level and quantity of lymphocytes which express fibronectin inside the cell and on cell surface was detected. Results of this investigation demonstrate that fibronectin-1 mRNA expression in lymphocytes is disturbed in erythremia disease and is accompanied by a decrease of fibronectin plasma level.

Journal Article
TL;DR: The paper is focused on the primary analysis of the potential genes of primary response to IFNalpha, predicted by the program COTRASIF elaborated in the laboratory, and the enriched category "Immune response" was detected with Mbl1 gene, which is reckoned among "unknown" genes.
Abstract: The paper is focused on the primary analysis of the potential genes of primary response to IFNalpha, predicted by the program COTRASIF elaborated in our laboratory. Two web-instruments FatiGO and GOTM were applied for this purpose; the literature search was carried out using IHOP; genes were classified according to the conceptual diagram "Gene Ontology" (GO) and were ranked according to their first priority for the experimental validation. On the basis of conducted analysis 162 genes of potential primary response to IFNalpha were subdivided into two groups--61 genes, for which the experimental data of their responsibility to IFNalpha were found and 101 genes for which such data are absent. We have performed the functional analysis of the "unknown" genes according to GO. The enriched category "Synapse part" encountering three genes of Rattus norvegicus: PRKCA-binding of protein, NMDAR-L and GABAA--receptor subunit alpha-2 were revealed. Among 162 genes the enriched category "Immune response" was detected with Mbl1 gene, which is reckoned among "unknown" genes. The four designated genes are chosen as the candidates for the prior validation.

Journal Article
TL;DR: It has been shown that calix[4]arene C-99 inhibited myosin subfragment-1 ATPase of myometrium, and this compound reduces the seeming enzymatic hydrolysis maximum rate of nucleoside triphosphate with respect to ATP and Mg2+.
Abstract: It has been shown that calix[4]arene C-99 inhibited myosin subfragment-1 ATPase of myometrium. This inhibition is noncompetitive as to ATP and Mg2+. At the same time, this compound reduces the seeming enzymatic hydrolysis maximum rate of nucleoside triphosphate with respect to ATP and Mg2+. With the help of computer design the interaction of mentioned calix[4]arene with myosin subfragment-1 of myometrium has been investigated. Several mechanisms involved in the calix[4]arene C-99 inhibition of myosin head ATPase were supposed and participation of hydrogen, hydrophobic and electrostatic interactions in these mechanisms was discussed.

Journal Article
TL;DR: The content of vitamins E, A1, A2 and carotenoids in the liver of fishes of different species at the beginning of winter and spring was significantly lower than at the start of summer and autumn and species differences in antioxidant enzymes activity are insignificant.
Abstract: The content of lipid peroxidation products--diene conjugates, lipid hydroperoxides, thiobarbituric acid reactive substances (TBARS), vitamins A, E and carotenoids and the activity of antioxidant enzymes--superoxide dismutase, glutathione peroxidase and catalase in the liver of freshwater fishes of different species (silver carp, grass carp and common carp) in different seasons have been studied. It was established the activity of antioxidant defence system in the liver of fish depends significantly on the season and fish species. In particular, the content of lipid peroxidation products in the liver of freshwater fishes at the beginning of winter and spring was significantly higher compared to their content at the beginning of summer and autumn. The superoxide dismutase and glutathione peroxidase activities in the liver of these fish species at the beginning of winter and spring were significantly lower than at the beginning of summer and autumn while the seasonal changes of catalase activity in the liver of fish are expressed insignificantly. The content of vitamins E, A1, A2 and carotenoids in the liver of fishes of different species at the beginning of winter and spring was significantly lower than at the beginning of summer and autumn. The content of lipid peroxidation products and vitamins E, A1 and A2 in the liver of common carp is significantly lower than in the liver of silver carp and grass carp and species differences in antioxidant enzymes activity are insignificant.

Journal Article
TL;DR: It was shown that two small RNAs about 65 and 55 nucleotides long included in NPV B form with polypeptides p29 and p14 specific RNP-complexes with molecular weights of 50 and 31 kDa, respectively.
Abstract: It was shown that two small RNAs about 65 and 55 nucleotides long included in NPV B. mori polyhedra form with polypeptides p29 and p14 specific RNP-complexes with molecular weights of 50 and 31 kDa, respectively. Both complexes form high-molecular weight complex with polyhedrin. Origin and nature of p29 and p14 polypeptides are discussed.

Journal Article
TL;DR: The maleimide derivative prevents development of oxidation stress and partially reduce them to control level in liver antioxidant system and level of matrix metalloproteinase-2 in intestinal mucosa after chronic treatment with 1.2-dimethylhydrazine-induced colon cancer in rats.
Abstract: The maleimide derivative--1-(4-Cl-benzyl)-3-Cl-4-(CF3-phenylamino)-1H-pyrrol-2.5-dione (MI-1) with cytostatic activity did not cause substantial changes of liver antioxidant system and level of matrix metalloproteinase-2 in intestinal mucosa after chronic treatment (for 20 weeks). MI-1 did not cause significant changes in the content of thiobarbituric-active products and plasma membrane protein carbonyl groups in the rat liver. However activities of superoxide dismutase, glutathione peroxidase, and content of reduced glutathione were decreased in both doses--0.027 and 2.7 mg/kg. The level of matrix metalloproteinase-2 in intestinal mucosa was decreased just in maximum dose--2.7 mg/kg. The contents of thiobarbituric-active products, protein carbonyl groups, reduced glutathione, matrix metalloproteinase-2, activities of glutathione peroxidase and glutathione-S-transferase in the liver cells have increased in 1.2-dimethylhydrazine-induced colon cancer in rats. The activities of enzymes of the first line of antioxidant defense--superoxide dismutase and catalase were decreased to 40%. The maleimide derivative prevents development of oxidation stress and partially reduce them to control level.

Journal Article
TL;DR: Data can evidence for the presence of correlation between the age of patients, latency period and induction of RET/PTC3 oncogenes.
Abstract: A comparative analysis of the expression of both, RET/PTC1 and RET/PTC3 oncogenes in papillary thyroid carcinomas (PTC) of patients from different age groups was carried out. Those were the following groups: children (mean age - 13 years, mean latency period - 13 years), young adults (mean age - 24 years, mean latency period - 14 years), adults (mean age - 38 years, mean latency period - 22 years). The presence of RET/PTC oncogenes was detected using polymerase chain reaction. In all cases the samples of both tumor and normal thyroid tissue were studied. It was established that induction of both, RET/PTC1 and RET/PTC3 rearrangements was present only in carcinoma samples. In PTCs the percentage of RET/PTC-positive tumors with increasing the age of patients has been decreasing. It should be noted that the part of carcinomas with induction of RET/PTC1 did not change with increasing the age of patients. At the same time the frequency of RET/PTC3 rearrangements with the increasing both the latency period and age of patients, significantly decreased. In conclusion, our data can evidence for the presence of correlation between the age of patients, latency period and induction of RET/PTC3 oncogenes.

Journal Article
TL;DR: The obtained results have proved that the model calculated and optimized by us can be used for studing the mechanisms of interaction of the active center of the enzyme with substrates and inhibitors.
Abstract: A comparison has been made between spatial structures of human CYP2E1 obtained experimentally and those calculated using the computer methods. The structures were characterized by such parameters as total energy, protein pocket volume and total volume of molecules as well as the analysis of spatial geometry. The obtained results have proved that the model calculated and optimized by us can be used for studing the mechanisms of interaction of the active center of the enzyme with substrates and inhibitors. An assumption was made in the course of the research that one of the possible mechanisms of inactivation of the enzyme is the reduction of protein pocket volume, which prevents substrate access to the active center.

Journal Article
TL;DR: It is demonstrated clearly that silver nanoparticles significantly affect the expression of PFKFB-2 mRNA on the alternative splicing level in different vital organs and show their effect on the important mechanisms of metabolism regulation in the cells on the level of key enzyme gene expression.
Abstract: Bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-2 (PFKFB-2) is represented by several alternative splice variants and plays a significant role in the glycolysis regulation in the brain, lung, testis and heart cells. The expression of PFKFB-2 mRNA and its alternative splice variants in these rat vital organs after single intratracheal injection of silver nanoparticles was studied. It was shown that the expression of PFKFB-2 mRNA is significantly changed in different rat tissues under silver nanoparticles action. The effect of silver nanoparticles on the expression of PFKFB-2 mRNA was observed one day after its injection to animals. In 3 and 14 days the effect of silver nanoparticles was increased (in testes) or kept on the approximately same level (in other investigated tissues). The expression of PFKFB-2 mRNA in most tissues is returned to its control levels one year after the injection of silver nanoparticles to the rats. It was also shown that the expression of alternative splice variants of PFKFB-2 mRNA without functional activity of 6-phosphofructo-2-kinase is significantly increased in different tissues 1, 3 and 14 days after single injection of silver nanoparticles. The results of this investigation demonstrate clearly that silver nanoparticles significantly affect the expression of PFKFB-2 mRNA on the alternative splicing level in different vital organs and show their effect on the important mechanisms of metabolism regulation in the cells on the level of key enzyme gene expression.

Journal Article
TL;DR: In this article, it was shown that activation of mitoKATP from the rat myometrium causes the increase of the hydrodynamic diameter of organelles and partial depolarization of the inner membrane.
Abstract: Mitochondrial ATP-sensitive potassium channel (mitoKATP) is a main factor of regulation of K+ exchange in mitochondria. Using photon correlation spectroscopy we have shown diazoxide-induced increase of hydrodynamic diameter of mitochondrial particles from the rat myometrium. Selective channel blocker glybenclamide partially eliminates this effect. By means of Rhodamine-123 fluorescence it was shown that activation of ATP-sensitive K(+)-channel in mitochondria caused partial depolarization of the mitochondrial membrane. This effect was absolutely blocked by glybenclamide. In the presence of valinomycine and diazoxide together, depolarization also was detected, but in this case glybenclamide failed to restore mitochondrial potential. Thus, activation of mitoKATP from the rat myometrium causes the increase of the hydrodynamic diameter of organelles and partial depolarization of the inner membrane.

Journal Article
TL;DR: The conclusion was made that oxytocin does not influence the passive Ca2+ efflux and it was proved that hTrpC channels take part in the regulation of free Ca ions concentration in the myometrium cells.
Abstract: The work deals with the study of molecular and membrane mechanisms of uterotonic peptide hormone oxytocin action on Ca ions homeostasis in the uterus myocites and activity of passive and active transport systems of this cation, localized in the myometrium subcellular structures. Biochemical mechanisms of additional Ca2+ influx into the myometrium cells from extracellular environment activated by oxytocin and thapsigargin were determined. Such Ca2+ influx has got the name of capacitative cation entry (CCE), or which is activated by intracellular store depletion (store-operated calcium entry). It was shown that cells from the nonpregnant human myometrium and PHM1-41 cells (immortalized pregnant human myometrial cells) expressed endogenous hTrpC1, 3, 4, 6 and 7 mRNA. The membrane-permeable derivative of diacylglycerol (OAG) stimulated an increase in oscillations of intracellular free Ca2+ concentration in PHM1-41 and myometrium cells. OAG-induced Ca2+ -oscillations were not affected by inhibition of PKC. It was proved that hTrpC channels take part in the regulation of free Ca ions concentration in the myometrium cells. On the basis of results of experiments, conducted on myometrium plasma membrane fraction, the conclusion was made that oxytocin does not influence the passive Ca2+ efflux. It was shown that peptyde hormone partially inhibited Ca2+ accumulation in plasma membrane fraction. Oxytocin also partially inhibited endoplasmic reticulum calcium pump activity of the myometrium cells. The conceptual pattern of myometrial Ca2+ exchange regulation by oxytocin is offered.

Journal Article
TL;DR: The survey considers the involvement of hormones, cytokines and growth factors in HIF stimulation under normoxia and special attention is given to the role of phosphorylation and other post-translational modifications in the regulation of HIF expression and activity.
Abstract: The review is devoted to the role of hypoxia-inducible factors (HIF) in the regulation of oxygen-dependent gene signalling. Structural features of HIF alpha and beta subunits as well as involvement of hydroxylation in the regulation of HIF stability and activity are described. Special attention is given to the role of phosphorylation and other post-translational modifications in the regulation of HIF expression and activity. The survey considers the involvement of hormones, cytokines and growth factors in HIF stimulation under normoxia. HIF target genes and promotor/enhancer sequences, responsible for oxygen-dependent gene regulation, are also described.

Journal Article
TL;DR: In this review a short history of these enzymes investigation, classification, structure, and functional significance of ecto-nucleoside triphosphate diphosphohydrolases (E-NTPDase) has been presented.
Abstract: Ecto-nucleotidases are enzymes of hydrolase class. They split extracellular nucleoside tri- and diphosphate. In this review a short history of these enzymes investigation, classification, structure, and functional significance of ecto-nucleoside triphosphate diphosphohydrolases (E-NTPDase) has been presented. These enzymes are glycoproteins anchored in membranes. They do not form phosphorylated enzyme's form during catalytic circle, and (by analogy with membrane-bound ATPases) form homooligomeric ensembles. Activity of these enzymes depends on bivalent ions, in particular Ca2+ and Mg2+. E-NTPDases function in the composition of ecto-nucleotidase cascade that contains other nucleotide-hydrolyzing enzymes. They regulate P2-receptors by hydrolyzing its ligand specifically ATP. Both modern information and results of our investigation about influence of different endo- and exogenous factors on activity of these enzymes has been presented.

Journal Article
TL;DR: The available data, including own researches indicate the Na+,K(+)-ATPase involvement in certain pathologies that develop with aberration of polarized epithelial phenotype, malignant growth in particular.
Abstract: The novel views on fundamental role of Na+,K(+)-ATPase in regulation of the polarized morphology of the epithelial cells, enzyme involvement in signal transduction, assembly of intermolecular signaling complexes have been reviewed. The modern state of researches are considered in detail, including: sorting specificity in polarized cells, Na+,K(+)-ATPase involvement in epithelial cell interactions, enzyme arrangement in combined structural and functional ensemble with adhesion and cytoskeleton proteins, their coordinated exp ression. The mechanisms of the Na+,K(+)-ATPase association with its molecular partners and the role of specific domains, motives and binding sites are analysed. The available data, including own researches indicate the Na+,K(+)-ATPase involvement in certain pathologies that develop with aberration of polarized epithelial phenotype, malignant growth in particular.

Journal Article
TL;DR: Studies of differential expression and activity of PKD1, PKD2 and PKD3 in the context of tumor spreading and prognosis could be the perspective subject of translational research in oncology.
Abstract: Recent scientific research demonstrates that protein kinases of PKD family affect biological features of normal and malignant cells. Differential expression of PKD genes was found in tumors of different histogenesis. These protein kinases could be activated by growth factors, antigen stimulation, and oxidative stress, the processes that usually can be observed during tumor progression. PKD regulate cell-cell contacts by affecting cell adhesion. PKD are involved in the regulation of cell proliferation and apoptosis, and also participate in epigenetic regulation of gene expression. That is why studies of differential expression and activity of PKD1, PKD2 and PKD3 in the context of tumor spreading and prognosis could be the perspective subject of translational research in oncology. This will contribute to the development of new approaches to differential diagnostics of tumor and target therapy, and also reveal prognostic factors for the prediction of clinical outcome.