New Aspects in Molecular Biotechnology and Biochemistry 2013
New Aspects in Molecular Biotechnology and Biochemistry 2013
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New Aspects in Molecular Biotechnology and Biochemistry 2013
Young Scientists' Conference
NEW ASPECTS IN MOLECULAR BIOTECHNOLOGY AND BIOCHEMISTRY
27-28 June, 2013
H. Buniatian Institute of Biochemistry NAS RA
Yerevan, Republic of Armenia
ABSTRACTS
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New Aspects in Molecular Biotechnology and Biochemistry 2013
Contents:
RGANIZING COMMITTEE 5
PREFACE 6
DETECTION AND QUANTIFICATION OF PRP-1 IN BIOLOGICAL FLUIDS BY 7 USING ANTI-PRP-1 POLYCLONAL ANTISERUM Abrahamyan S.S., Khachatryan A.R., Tumasyan N.V., Sahakyan I.K. , Harutyunyan H. , Davtyan T.K.
GENDER DIFFERENCES OF PARP-1 ACTIVITY IN LIVER 8 Asatryan A.L., Artsruni I.G., Matinyan K.S., Margaryan A. V., Gevorgyan E.S.
CHANGES IN THE ARGINASE ACTIVITY IN MICE ASCITES FLUID AND BLOOD 9 IN EHRLICH ASCITES CARCINOMA FOLLOWING TREATMENT WITH NON- PATHOGENIC STRAINS OF ESCHERICHIA COLI Avagyan H.Kh.
STUDY OF MUSHROOMS` INTRACELLULAR EXTRACTS ANTI- 10 INFLAMMATORY AND ANTICANCER ACTIVITY Avagyan I.A. , Minasbekyan L.A., Nanagulyan S.G.
CHANGES IN APOPTOTIC RATE AND SYNAPTIC PLASTICITY IN PATIENTS 11 WITH POSTTRAUMATIC STRESS DISORDER Avetyan D.
INVESTIGATION OF THE ANTIBACTERIAL ACTIVITY OF PROLINE-RICH 12 POLYPEPTIDES AGAINST BACILLUS ANTHRACIS Badalyan A.M., Badalyan Kh. V.
REGENERATION OF BURN INJURY UNDER THE INFLUENCE OF OINTMENTS 13 FROM THE EXTRACTS OF MULBERRY LEAVES AND SILVER BERRY SEEDS Balasanyan M.G., Soghbatyan L.T., Grigoryan D.S.
SOME PROPERTIES OF ANTIBACTERIAL COMPONENT OF LACTIC ACID 14 BACTERIA ISOLATED FROM ARMENIAN DAIRY PRODUCTS Bazukyan I., Babayan A., Trchounian A.
DO WE HAVE A CHANCE TO TEST ADEQUATELY NEWLY DEVELOPED 15 DRUGS IN ANIMAL MODELS OF FOCAL ISCHEMIA? Danielyan K.E.
NEW METHOD FOR PURIFICATION OF XANTHINE DEHYDROGENASE 16 Feresheryan K, Manucharyan T. G, Gyongyan S.A., Danielyan K.E.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
17 QUANTUM CHEMICAL STUDY OF DIMETHYL-AND DIETHYLSULFONES Gabrielyan L.S., Mkhitaryan A.S., Markarian S.A.
ELECTROPHYSIOLOGICAL STUDIES OF LATERAL VESTIBULAR NUCLEUS 18 AFTER NUCLEOTIDE THERAPY OF DAMAGED SCIATIC NERVE Gevorgyan L.R., Chavushyan V.A., Simonyan K.V.
THE INTERACTION BETWEEN GANGLERON AND HUMAN SERUM ALBUMIN: 19 FLUORESCENCE STUDIES Ghazaryan A. G., K. R. Grigoryan
ANNEXIN 11 EXPRESSION PATTERN IN SCHIZOPHRENIA 20 Ghazaryan H.
XO REGULATES PURINE CATABOLISM BY FEEDBACK MECHANISM 21 Gyongyan S.A., Manucharyan T. G, Danielyan K.E.
XANTHINE OXIDOREDUCTASE IS A KEY ENZYME OF PURINE CATABOLISM 22 REGULATION Gyongyan S.A., Manucharyan T. G, Danielyan K.E, Kevorkyan G.A., Chailyan S.G.
PRODUCTION OF CELLULASE BY THE HALOALKALIPHILIC STRAINS OF 23 STREPTOMYCES ISOLATED FROM SALINE-ALKALINE SOILS OF ARARAT PLAIN, ARMENIA Hakobyan A., Panosyan H., Trchounian A.
CREATINE/CREATINE KINASE SYSTEM IN THE CELLULAR MECHANISMS OF 24 EHRLICH ASCITES CARCINOMA AND THERAPY WITH NON-PATHOGENIC STRAINS OF ESCHERICHIA COLI Hovhannisyan M.R., Avagyan H.Kh., Movsesyan H.A., Alchujyan N.Kh., Movsesyan N.A.
TO METABOLISM OF TRANSMITTER AMINO ACIDS IN RAT BRAIN 25 Khachatryan N. Kh., Vardanyan A.G., Kamalyan R.G.
NEW METHOD FOR PURIFICATION OF XANTHINE OXIDASE 26 Manucharyan T. G., Gyongyan S.A., Danielyan K.E.
CATALYTIC ACTIVITY OF Mg²⁺- AND Ca²⁺-DEPENDENT ATPases IN 27 MITOCHONDRIAL TISSUES OF SOME SEVAN FISHES Margaryan A.S., Badalyan R.B., Simonyan A.A.
MEFV GENE EXPRESSION DURING MACROPHAGE ACTIVATION 28 Nersisyan L.R., Arakelyan A.A.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
FTIR STUDY OF THE REDOX PROPERTIES OF CYSTEINE IN THE PRESENCE OF 29 DIMETHYLSULFOXIDE Papanyan Z., Markarian S.
VOLUMETRIC PROPERTIES OF AOT/N-HEPTANE/DMSO-WATER REVERSE 30 MICELLAR SYSTEMS Shahinyan G.A., Sargsyan H.R., Markarian S.A.
PRODUCTION OF THERMOSTABLE ALPHA-AMYALSE BY BACILLUS SP. 31 IRANIAN S2 USING SOLID STATE FERMENTATION Sharifi Alghabpoor S., Panosyan H., Popov Yu., Trchounian A.
EXCITATION - EMISSION MATRIX FLUORESCENCE SPECTROSCOPY STUDIES 32 OF DIMETHYLSULFOXIDE EFFECT ON HUMAN SERUM ALBUMIN STABILITY Shilajyanand H. A., Grigoryan K. R.
ALTERATIONS OF LIPID MODIFICATION PROCESSES IN THE PERIPHERAL 33 BLOOD MONONUCLEAR CELLS IN MALIGNANCY Torgomyan T.R., Lazyan M.P., Hakobyan G.V., Batikyan T.B., Ghazaryan R.A., Alexanyan K.A., Galstyan H.M., Tadevosyan Y.V.
OXIDATIVE STRESS AND PATHOMECHANISMS OF ISCHEMIC STROKE 34 Tsakanova G.V., Ayvazyan V.A., Boyajyan A.S.
STUDY AND ASSESSMENT OF MICROBIAL COMMUNITIES IN NATURAL AND 35 COMMERCIAL BIOLEACHING PROCESSES Vardanyan A.K., Khachatryan A.N., Stepanyan S.Kh.
FRET MICROSCOPY FOR REAL-TIME MONITORING OF cGMP INDUCED BY 36 PRP-1 Yeranosyan L.A.
NEUROTROPHIN FAMILY GENE AS POTENTIAL TARGET 37 FOR SCHIZOPHRENIA Zakharyan R. V., Boyajyan A. S.
AB INITIO AND DFT THEORETICAL STUDY OF THE INTERACTION OF L- 38 AA/DESO Zatikyan A.L., Markarian S.A.
The abstracts are published in Autor's version. The Autors are the only who responsible for all the information published in articles.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
Organizing Committee
Samvel Chailyan Ph.D., D. Sci., Director of the H. Buniatian Institute of Biochemistry, Head of the Department of the Biochemistry of Neurohormones / URL: http://aab.sci.am Anahit Margaryan Ph.D Senior researcher, H. Buniatian Institute of Biochemistry NAS RA / URL: http://aab.sci.am Torgom Seferyan Ph.D Researcher, H. Buniatian Institute of Biochemistry NAS RA / URL: http://aab.sci.am
Flora Saruchanyan Ph.D Senior researcher, H. Buniatian Institute of Biochemistry NAS RA / URL: http://aab.sci.am Ovsanna Hunanyan Ph.D Researcher, H. Buniatian Institute of Biochemistry NAS RA / URL: http://aab.sci.am
Inessa Sahakyan Ph.D Researcher, H. Buniatian Institute of Biochemistry NAS RA / URL: http://aab.sci.am
Roksana Zakharyan Ph.D, Researcher, Institute of Molecular Biology NAS RA / URL: http://www.molbiol.sci.am/ Kristine Danielyan Ph.D Senior researcher, H. Buniatian Institute of Biochemistry NAS RA / URL: http://aab.sci.am
Scientific Program Committee: Guevork Kevorkian Professor, Head of the Department of Pathological Biochemistry of H. Buniatian Institute of Biochemistry NAS RA / URL: http://aab.sci.am Mikhayil Aghajanov Professor, Head of the Biochemistry Dept of Yerevan State Medical University/ URL: www.ysmu.am/ Naira Zakaryan Ph.D., Senior Reasercher at the Laboratory of Neuropeptides Biochemistry, H. Buniatian Institute of Biochemistry NAS RA / URL: http://aab.sci.am Hrachya Vardapetyan Professor, Doctor of biological sciences, Dean of the department of Medicine and Biology of Russian- armenian (Slavonic) University / URL: www.rau.am Emil Gevorgyan Doctor of Biological Sciences, Professor, NAS RA Associate member, Dean of the department of Biology / URL: http://www.ysu.am Hripsime Hayrapetyan Ph.D., Associated Professor, Senior Scientific Researcher / Secretary of the Special Scientific Council H. Buniatian Instituteof Biochemistry NAS RA / URL: http://aab.sci.am Zaven Karalyan Doctor of Biological Sciences, Professor, Head of Lab, Institute of Molecular Biology NAS RA / URL: www.molbiol.sci.am/
Technical Support Committee (H. Buniatian Institute of Biochemistry): Qeshishyan Z., Gevorgyan K., Margaryan A., Seferyan T.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
PREFACE
In 2013 the National Academy of Sciences of RA celebrates its 70th glorious anniversary. The Academy was established in 1943, when there was life and death battle during the Great Patriotic War II. About half million soldiers of Armenian descent were fighting in the battlefields. Hovsep Orbeli, the famous historian-archeologist arrived to Armenia and started the formation of the newly founded academy, becoming the first president of this institution. First it was a two rooms area, where two people were working: the president himself and a young scientist invited from Leningrad Hrant Batikyan, who later became the rector of the State University and the dean of Biological Faculty. By their efforts, step by step, in a very short period of time famous scientists were united and started to develop the institution, founding scientific research institutes within the Academy. Biochemical research in Armenia began in the department of biochemistry functioning in the structure of the Institute of Physiology, on the basis of which the Institute of Biochemistry was founded in 1961 by the efforts of Academician Hrachya Buniatian, who became the first director of the institute from the day of its foundation until his death. Two years later the Association of Armenian Biochemists was formed, which included all the scientists in charge of biochemical research in Armenia, and already in 2004 it became the plenipotentiary member of FEBS, in 2010 it was already in the structure of IUBMB. Today you, the young scientists, must continue through better scientific achievements the honor of rich biochemical school that was inherited by you from the famous Armenian scientists. I wish you meaningful reports and fruitful discussions. Armenian science of tomorrow is yours.
Guevork A. Kevorkian, Professor The President of Armenian Association of Biochemists Academician of the European Academy of Sciences and Art and Russian Medical and Technical Academy
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New Aspects in Molecular Biotechnology and Biochemistry 2013
DETECTION AND QUANTIFICATION OF PRP-1 IN BIOLOGICAL FLUIDS BY USING ANTI-PRP-1 POLYCLONAL ANTISERUM
Abrahamyan S.S., Khachatryan A.R., Tumasyan N.V., Sahakyan I.K. , Harutyunyan H. , 1Davtyan T.K.
H.Buniatian Institute of Biochemistry of NAS RA, Yerevan, Republic of Armenia 1Laboratory of Immunology and Virology, Armenicum Research Center,JSC Armenicum, Yerevan, Republic of Armenia [email protected]
The objective of the present work was to develop a method enough sensitive for the detection and quantification of the proline rich polypeptide (PRP-1 called galarmin) in rat and human biological fluids, including serum, plasma and cerebrospinal fluid. Galarmin consisting of 15 amino acid residues (AGAPEPAEPAQPGVY) is one of the new types of cytokines of the neurosecretory hypothalamus, the proline rich peptides, isolated by prof. Galoyan and coworkers from bovine neurohypophysis neurosecretory granules and synthesized in the form of a common precursor protein (neurophysin- vasopressin associated glycoprotein). At present, a wide spectrum of the galarmin biological activity has been revealed. Up to now not any method has been used by us for PRPs quantification. An alternative solid phase readout system for the detection of antigen–antibody reactions is the ELISA assay. In this study, using an anti-rabbit primary antibody against the synthetic galarmin we developed an enzyme linked immunosorbent assay (ELISA) for detection of PRP-1 and structurally similar compounds named d-15; d-Gx-NH2; Gx, as well as the competitive ELISA for quantification of galarmin in biological fluids. Data indicated that d-Gx-NH2 and Gx do not affect the detection and quantification of PRP-1 by this ELISA method. This observation and the crossreactivity percentage obtained that the antiserum recognises only d-15 among the galarmin related peptides. According to the analysis of the competitive ELISA the minimum detectable amount of galarmin in the fluid was 1.5 ng/ml at the appropriate condition chosen to be the best one for galarmin detection: quantity of the immobilized galarmin (25 ng/ml); anti-rabbit primary antibody against the galarmin (1:5000); anti-rabbit secondary antibody conjugated to peroxidase (1:10000), and extravidin (1:10000). Thus, an ELISA system has been developed using polyclonal antibody raised against the synthetic galarmin that demonstrated the sensitivity and the specificity of the method.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
GENDER DIFFERENCES OF PARP-1 ACTIVITY IN LIVER
Asatryan A.L., Artsruni I.G., Matinyan K.S., Margaryan A. V., Gevorgyan E.S.
Yerevan State University, Faculty of Biology, Yerevan, Armenia [email protected]
Poly(ADP-ribose)polymerase-1 (PARP-1) is abundant nuclear enzyme and the most elaborated member of PARP enzymes superfamily. The enzyme is implicated in regulation of chromatin structure and in a wide range of chromatin-associated processes, e.g. DNA replication, reparation, transcription [3] and PARP-1 activity significantly affects cellular responses to survival/death signals. The data revealed that PARP inhibition protects against injury in ischemia–reperfusion-associated and cardiovascular diseases, diabetes, stroke, etc [1,4]. Pharmacological inhibition of PARP-1 potentiates the effect of alkylating agents and ionizing radiation, and sensitizes cancer cells to chemo- and radiotherapy. Treatment with PARP-1 inhibitors down regulates inflammatory mediator production and reduced mortality in male, but not female rats [2]. While majority of investigations is focused on sexual dimorphism of PARP inhibition displayed by neuronal cells, we didn't succeed in revealing data relevant to gender differences in PARP-1 inhibition in different organs or cells. In present study we attempt to determine whether there are gender differences in PARP-1 activity in rat liver cells. Our experimental data indicate, that PARP-1 activity in liver nuclei of male rats is higher than that in female ones (nearly by 37%). To diminish the effects of blood-circulating sex hormones we have investigated ATP and bezamide (Bam) effect on PARP-1 activity in naked liver nuclei of rats of both sexes. Data accumulated show dramatic reduction of enzyme activity which was displayed by nuclei, and nearly a complete inhibition of PARP-1 was apparent at 5mM ATP for all investigated probes. Inhibitory effect of ATP didn’t exhibit sex-specific differences. The effect of the first generation of PARP-1 inhibitor Bam was also examined in rat liver nuclei. Results show that Bam inhibition displays significant gender differences in rat liver nuclei: it dramatically decreases enzyme activity in male, and by nearly 50% in female organisms.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
CHANGES IN THE ARGINASE ACTIVITY IN MICE ASCITES FLUID AND BLOOD IN EHRLICH ASCITES CARCINOMA FOLLOWING TREATMENT WITH NON-PATHOGENIC STRAINS OF ESCHERICHIA COLI
Avagyan H.Kh.
H.Buniatian Institute of Biochemistry NAS RA,Yerevan, Armenia [email protected]
The accumulated findings indicate the promising potential of non-virulent bacteria as cancer immunotherapeutic agents. Arginase has been shown to be either responsible for or to participate in tumor immune escape. We have studied the arginase involvement in the intracellular mechanisms of bacterial oncotherapy. On the 11-th day of Ehrlich ascites carcinoma (EAC) development the activity of the arginase isoforms (types I and II) increased for 7,7, and 12,3 times in cytoplasm and mitochondria of blood leukocyte, respectively, compared to control, whereas of about a two-fold increase in total arginase activity was observed in both leukocyte homogenate and plasma. At the less extent arginase I and II were activated to 5,2-, and 3 times, respectively in peritoneal leukocyte. Two days after i.p. injection with EAC cells, a single intraocular treatment by live non-pathogenic clinical strains of Escherichia coli exhibited a remarkable antitumor activity in EAC-bearing mice causing a three-fold decrease in the ascites fluid volume and of about 75 % prolongation the life span. At the same time, E coli-treatment increased thrice the arginase I activity, while that of arginase II diminished 2,5 times in blood leukocyte, but total arginase activity was not significantly changed in leukocyte homogenate and plasma. Conversely, arginase I activity decreased 1,4 times and arginase II was normalized in peritoneal leukocyte following E. coli-treatment. Moreover, arginase activity dropped 4,7 times in ascites fluid. It is of importance, because overexpressing either arginase I or II in cells may not only reduce NO synthesis but also can enhance polyamine synthesis and cell proliferation. Taken together our results suggest the modulatory effects of avirulent strains of E. coli on the arginase isoforms in immune cells from blood and peritoneal cavity, as well as down-regulation the arginase in tumor cells that should be taken into account in E. coli application in cancer therapy.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
STUDY OF MUSHROOMS` INTRACELLULAR EXTRACTS ANTI- INFLAMMATORY AND ANTICANCER ACTIVITY
Avagyan I.A. , Minasbekyan L.A., Nanagulyan S.G.
Faculty of Biology, Yerevan State University, Yerevan, Armenia, [email protected]
Numerous bioactive polysaccharides or polysaccharide-protein complexes from medicinal mushrooms are described that appear to enhance innate and cell-mediated immune responses and exhibit antitumor activities in animals and humans. Stimulation of host immune defense systems by bioactive polymers from medicinal mushrooms has significant effects on the maturation, differentiation, and proliferation of many kinds of immune cells in the host. Many of these mushroom polymers were reported previously to have immunotherapeutic properties by facilitating growth inhibition and destruction of tumor cells. While the mechanism of their antitumor actions is still not completely understood, stimulation and modulation of key host immune responses by these mushroom polymers appears central [1]. We modulate growth conditions of mushroom culture, which lead to the sharp increasing of peroxidase activity of up to 300% and betta–glucosidase up to 200% at the some frequencies, as well as obtained increasing of protein content in extracts [2, 3]. The purpose of this work is to determine the anti-inflammatory activity of three wood- decaying mushrooms ‘cultures extracts on a widely used model of rat ear acute inflammation, induced by xylol. Intraperitoneal injection of an extracts from the irradiated by mm-waves cultures of the mushrooms are suppress an acute inflammation by 85 %. Moreover we have possibility investigated and anticancer activity of extracts on the different carcinoma tissues cells in vitro. As evidence our results extracts from culture of some wood-decaying mushrooms possessive by antiproliferative activity, by suppressing mitotic activity of cells of some carcinoma tissues. On the base of obtained results we suggest that immune reply of the body (rat or else) at the treatment of inflammation and cancer by the mushrooms extracts has a same mechanism.
1. Wasser SP Current findings, future trends, and unsolved problems in studies of medicinal mushrooms.//Appl Microbiol Biotechnol. 2011 Mar; 89(5):1323-1332. 2. Minasbekyan L A, Nanagyulyan S G, Avagyan I A (2009) Increase of betta-glycosidase activity of P. ostreatus culture in response to the stress. Immunopathology, allergology, infectology, 1, 26-27. 3. Avagyan I.A., Nerkararyan A.V., Minasbekyan L.A., Nanagulyan S.G. (2011) Influence of mm-waves on growth and fermentative activity of Pleurotus ostreatus mushroom culture. Micologiya i Phytopatologiya, 6, 77- 83.
Acknowledgments. This work was made possible be research grants: from the Armenian National Science and Education Fund (ANSEF) based in New York USA, plant-3261 and National State Committee on Science of the Ministry of Education and Science of the Republic of Armenia (13A-1g29).
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New Aspects in Molecular Biotechnology and Biochemistry 2013
CHANGES IN APOPTOTIC RATE AND SYNAPTIC PLASTICITY IN PATIENTS WITH POSTTRAUMATIC STRESS DISORDER
Avetyan D.
Institute of Molecular Biology NAS RA, Yerevan, Armenia [email protected]
Posttraumatic stress disorder (PTSD) is severe polygenic psychiatric disease characterized by cognitive impartment, which may result from apoptotic and synaptic plasticity (SP) dysfunction. In the present study blood levels of annexin-a5 and complexin-2 proteins involved in apoptosis and SP were determined and functional single nucleotide polymorphisms (SNPs) of genes encoding these proteins (ANXA5 and CPLX-2) were evaluated in patients with PTSD (DSM-IV-TR code: 309.81) in comparison to healthy subjects (HS). In total, 100 patients with PTSD (Karabakh combat veterans) and 100 HS were involved in this study. The study was approved by the Ethical Committee of the Institute of Molecular Biology NAS RA (IRB #00004079). The experiments were performed using blood serum/plasma and genomic DNA samples of study subjects. Methodological design was based on the enzyme-linked immunosorbent assay and polymerase chain reaction with sequence- specific primers. Data were evaluated using Hardy-Weinberg equilibrium, Pearson’s Chi-square test, Bonferroni multiple correction approach, Mann-Whitney U test, Kruskal-Wallis H-test, Dunn's multiple comparison test and Spearman correlation analysis. The obtained results indicate that: (1) PTSD is characterized by hypoactivity of apoptosis manifested by decreased blood levels of annexin-a5; (2) PTSD is characterized by decreased SP manifested by decreased plasma levels of complexin-2; (3) alterations in apoptosis rate and SP in PTSD are interrelated; (4) PTSD is associated with the SNP rs1366116 of CPLX-2; (5) the rs1366116*T mutant allele of CPLX-2 represents risk factor for PTSD; (6) decreased blood levels of complexin-2 protein in PTSD result from a prevalence of the rs11575945*T mutant allele of CPLX-2 in PTSD-affected subjects. Alterations in apoptotic rate and SP are involved in pathogenesis of PTSD. Annexin-a5 and complexin-2 proteins blood levels may be considered as molecular markers of altered apoptosis and SP in PTSD. CPLX-2 may be nominated as a candidate gene for PTSD.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
INVESTIGATION OF THE ANTIBACTERIAL ACTIVITY OF PROLINE- RICH POLYPEPTIDES AGAINST BACILLUS ANTHRACIS
Badalyan A.M., Badalyan Kh. V.
H. Buniatian Institute of Biochemistry NAS RA,, Department of Neurohormones Biochemistry, Laboratory of neurochemistry and neuroimmunology of special dangerous infectious diseases, Yerevan, Armenia [email protected]
Anthrax induces acute infectious disease. Currently, treatment for anthrax infection involves the use of several different antibiotics. But now there are any strains of bacillus anthracis, which are resistant to currently used antibiotics. The experiments were carried out in vi vo on mi ce model to show the protective and antibacterial effects of new hypothalamic proline-rich polypeptides (PRPs) against anthrax . Prior to testing the influence of galarmin, it was necessary to determine the minimal lethal dose of Bacillus anthracis in whit e mi ce . All the mice were infected i.p. with anthrax strain N55 vaccine (1x107/mice). The cont rol group received vehicl e (0. 4 ml of 9% NaCl, i.m.); experimental animals received different concentrations of galarmin (single administration of galarmin is 16mkg on the infection day, and repeated in 24-72h, i.m.). The mortality and rate of survival was visually observed during the 10th day. The experiments showed that after infectioning by the most virulent bacilli, the control animals died within 3-6 days with development of all typical symptoms of the disease. In the group of galarmin-treated mice the rate of survival increased by 60% compared to non-treated (vehicle control) mice. Pathological, anatomic and microbiological analysis showed that the animals died because of anthrax. Usually, after vaccination of animals the organisms vaccine bacteria continue to develop in the organism and produce an environment during about 100 days. The results of experiments show that under the influence of galarmin the elimination period of bacilli is significantly reduced from the organism. The results of experiments show that the hypothalamic neurosecretoiry cytokines consisting of 15 amino acid residues manifest a strong prophylactic and therapeutic effect towards animals infected by anthrax.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
REGENERATION OF BURN INJURY UNDER THE INFLUENCE OF OINTMENTS FROM THE EXTRACTS OF MULBERRY LEAVES AND SILVER BERRY SEEDS
Balasanyan M.G., 1Soghbatyan L.T., 2Grigoryan D.S.
YSMU after Mkhitar Heratsi, Department of Pharmacology, Yerevan, Armenia YSMU after Mkhitar Heratsi, Department of Pharmacy, Yerevan, Armenia 2 Scientific Centre of Radiation Medicine and Burns, Yerevan, Armenia [email protected] , 1 [email protected], 2 [email protected]
It is well known that thermal trauma leads to formation of free radicals. Ascorbic acid and flavonoids having free hydroxyl groups in their molecules easily oxidize after interaction with free radicals, thus showing high scavenging activity and appear significant antioxidant activity. That is why high tendency of treatment of burn injury with preparations of plant origin is noted recently due to high concentration of flavonoids, phenolic compounds, ascorbic acid and saponins. Literature data and our latest experiments show that mulberry leaves and silver berry seeds have rich content of flavonoids, ascorbic acid, saponins and phenolic compounds. Taking into account all the mentioned facts, the regeneration ability of the ointments from the extract of mulberry (Morus alba) leaves (I ointment) and from the combined extract of the latter with the silver berry (Elaeagnus angustifolia) seeds (II ointment) was studied under the conditions of thermal burn. Experiments were done on albino imbread rats, weighing 180-200g. Estimation of the ointments burn injury regeneration activity was carried out by the evaluation of burn surface area changes in the control and experimental groups by the modified method Garros et al. The carried out experiments evident that the area of burn surface was decreased on the 3rd , 7th, 14th days after inducing thermal burn by 6,65% and 5,73%, 8,07% and 7,43%, 41,75% and 6,4% consequently applying I and II type of ointments, compared with control group. It has been found out that the ointments reduce pH of burn surface with significant decrease in microbial and fungi flora. Obtained data evident that pharmaceuticals based on the extracts of mulberry leaves and silverberry seeds could be useful for treatment of several type tissue injuries.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
SOME PROPERTIES OF ANTIBACTERIAL COMPONENT OF LACTIC ACID BACTERIA ISOLATED FROM ARMENIAN DAIRY PRODUCTS
Bazukyan I., Babayan A., Trchounian A.
Yerevan State University, Faculty of Biology, Department of Microbiology and Biotechnology of Plants and Microorganisms , Yerevan, Armenia [email protected]
The five LAB strains with high antibacterial activity have been isolated from matsoun and cheeses of Armenia. The all strains demonstrated the cytostatic antibacterial activity against both G+ and G- bacteria (diameter of test-microorganisms growth suppression zones 14-30 mm). The synthesis of antibacterial components started in the first part of idiophase and the maximal synthesis of antibacterial component observed after 33 hours of cultivation in MRS for INRA- 2010-5.2 and INRA-2010-21.2 and after 40 hours for the other strains. The minimal inhibitory concentrations of cultural liquid were 1:2.5, 1:1, 1:1, 1:1.5 and 1:1.5 for the strains INRA-2010- 4.2, INA-5.1, INA-21.1, INRA-2010-5.2 and INRA-2010-21.2, correspondingly. The inhibitory effect of pH on the antibacterial activity of INA-5.1 and INA-21.1 strains was observed at pH 6.5, for INRA-2010-4.2 strain at pH 8.0; the antibacterial activity of the strains INRA-2010-5.2 and INRA-2010-21.2 were stable at pH 8.0. Although antibacterial components showed high stability after treatment at 85oC against Salmonella typhimurium MDS-1754, they were not stable after treatment at 60-85oC against Streptococcus aureus MDS-5233. It was revealed the inhibitory effect of oxygen. The cultural liquids of INA-5.1, INRA-2010-5.2 and INRA-2010- 21.1 lost antibacterial activity after ultrasound treatment. The treatment with proteinase K partially decreased the antibacterial activity of all strains except INRA-2010-21.2, which completely lost activity, so the antibacterial components could be protein like. All strains were resistant to 0.3% phenol, but the most of them lost the antibacterial activity after treatment. Thus, all the studied strains suggested can be used as starters for production of functional food, as well as probiotics or preserving strain-producers.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
DO WE HAVE A CHANCE TO TEST ADEQUATELY NEWLY DEVELOPED DRUGS IN ANIMAL MODELS OF FOCAL ISCHEMIA?
Danielyan K.E.
H. Buniatian Institute of Biochemistry NAS RA, Yerevan, Yerevan, Armenia [email protected]
Animal models of ischemic stroke, hemorrhagic transformation certainly has vivid importance for stroke research, development of thrombolytic as well as neuroprotective drugs. Clear understanding of techniques for every type of the stroke models highlights naturally impossible adverse effects of the surgery, which might greatly influence on interpretation of final experimental results. There is not any stroke model, which will fully reflect human disease. Infarcts are relatively larger in experimental animals than in humans with strokes. The models are more analogous to massive hemispheric infarcts than to localized strokes such as those in the internal capsule. Every type of the animals stroke models is a partial hallmark of clinical picture. Thus, knowledge of variety of stroke models allows choosing the system, which will allow adequately testing drugs or compound, predicting effective doses, and evaluating possible adverse effects, pharmacokinetic.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
NEW METHOD FOR PURIFICATION OF XANTHINE DEHYDROGENASE
3Feresheryan K., 1, 2Manucharyan T. G., 1Gyongyan S.A., 2Danielyan K.E.
1Department of Biochemistry, Yerevan State University, Armenia 2H. Buniatian Institute of Biochemistry, NAS, Armenia 3Slavonic University, Yerevan, Armenia Contact person: [email protected]
Lyon E.S. with colleages purified the XO/XDH (Xanthine Oxidase; EC 1.3.22 and Xanthine Dehydrogenase; EC 1.17.1.4) from the Neurospora crassa by the utility of immunoadsorbing methods. Mono specific antibodies were conjugated with the Sepharose 6B. Electrphoresis has visualized just one band of the proteins possessing with XDH activity1. Rajagopalan K.V. has purified XDH from the liver of the chickens by the utility of protein denaturation methods with further application of the DEAE- sephacell and G-200 chromatography 2. We are presenting here the new method of XDH purification with the utility of preparative electrophoresis. Activity of XDH was evaluated based on the method introduced by Litwack et all3. It was developed the new method based on the existing one 4, for the purification of XDH from the liver of rats with the final utility of the preparative electrophoresis with further extraction of the proteins from the gel and evaluation of XDH activity. For the calculation of the statistic we have used ONE-WAY-ANOVA (results were considered significant, when p<0.05) There were visualized by phoresis two bands possessing with the XDH activity. One of the bands had an approximate molecular weith equal to 300 kDa, the other one had approximately 40 kDa molecular weight. We have delineated also the activity of the XO in the pure fractions. Activity was detected in the first as well as 5th fractions (0.5156±0.0356- control, 0.9333±0.0889 -xanthine, 0.6904±0.0448-allopurinol p<0.01 for first fraction and 1.2125 ±0.5718 1.5229 ±0.1468, 1.0088±0.2134 for 5th fraction). We have developed new method of XDH purification and found out the XDH activity in 2 different fractions with 10 times difference in molecular weights.
References: 1. Lyon, E. S.; Garrett, R. H., Regulation, purification, and properties of xanthine dehydrogenase in Neurospora crassa. J Biol Chem 1978, 253(8), 2604-14. 2. Rajagopalan, K. V.; Handler, P., Purification and properties of chicken liver xanthine dehydrogenase. J Biol Chem 1967 242(18), 4097-107. 3. Litwack, G.; Bothwell, J. W.; Williams, J. J.; Elvehjem, C. A., A colorimetric assay for xanthine oxidase in rat liver homogenates. J Biol Chem 1953, 200, (1), 303-310. 4. Engerson, T. D.; McKelvey, T. G.; Rhyne, D. B.; Boggio, E. B.; Snyder, S. J.; Jones, H. P., Conversion of xanthine dehydrogenase to oxidase in ischemic rat tissues. J Clin Invest 1987, 79(6), 1564-70.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
QUANTUM CHEMICAL STUDY OF DIMETHYL- AND DIETHYLSULFONES
Gabrielyan L.S., Mkhitaryan A.S., Markarian S.A.
Yerevan State University, Department of Chemistry, Yerevan, Armenia [email protected]
Dimethyl sulfone (DMSO2) is an organic sulfur-containing compound that occurs naturally in a variety of fruits, vegetables, grains, and animals including humans. It has been suggested, that DMSO2 has an anti-inflammatory, antioxidant and chemopreventive mechanisms of action. The scientific interest has not only DMSO2 but also its nearest homologues diethyl sulfone (DESO2). Theoretical studies can help to develop an understanding of the mechanisms of their biomedical actions. Nowadays, quantum chemical calculations are able to provide very accurate predictions of molecular properties and energetics. In this work ab initio (HF, MP2) and density functional theory (DFT) methods with various basis sets are used to estimate thermodynamic properties of stable structures of DMSO2 and DESO2, and to predict the IR and Raman spectra of these molecules. Detailed vibrational assignments for DMSO2 and DESO2 have been done. It was shown, that with the basis sets of the 6-31+G(d) quality, the DFT calculated bond parameters and harmonic vibrations are in a very good agreement with experimental data. The calculations were carried out by using Gaussian 03 quantum chemistry program package. This work was carried out thanks to a research grant (PS-Cheminorg-3179) granted by the Armenian National Science and Education Fund (ANSEF).
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New Aspects in Molecular Biotechnology and Biochemistry 2013
ELECTROPHYSIOLOGICAL STUDIES OF LATERAL VESTIBULAR NUCLEUS AFTER NUCLEOTIDE THERAPY OF DAMAGED SCIATIC NERVE
Gevorgyan L.R., Chavushyan V.A., Simonyan K.V.
Orbeli Institute of Physiology NAS RA, Yerevan Armenia
Cell membrane damages and disorders of synthesis of membrane phosphatides play an important role in pathophysiology of peripheral nerves. Studies have shown that there is an increase in the needs of pyrimidine nucleotides after nerves injury. Alteration in phosphatidylcholine synthesis have been recognized as one of the mechanisms promoting the signaling cascade of apoptosis. The aim of the study was to estimate the neuroprotective effectiveness of Nucleo CMP (contains cytidine monophosphate and uridine triphosphate nucleotides) after unilateral compression of rat’s sciatic nerve (SN). We have recorded extracellular spike activity of single neuron of lateral vestibular nucleus (LVN) by stimulation of distal portion of the compressed SN. In Nucleo CMP group dominate neuronal units with inhibitory responses in the neurons of LVN. The comparative analysis of functional indices in intact and injured lower extremities revealed the recovery of motor and sensory function on the 30 day under action of Nucleo CMP. We have recorded the dynamics of spiking activity in single neurons (n=6) of LVN on intact rats before and after i/m administration a therapeutic doses of Nucleo CMP. Averaged data showed the changes of frequency of spike activity in neurons of the LVN neurons by stimulation of SN. Slowing of background and an increase of post-stimulus spiking in neurons of the LVN neurons was recorded within 15-80 minutes of Nucleo CMP action. Evidently, enhances the release of neurotransmitters, influencing on modulation of ion channels, sensitization/desensitization of glutamate and GABA postsynaptic receptors. Thus, the stimulating effect of Nucleo in excitatory and inhibitory neurotransmitters’ system is an important component of neuronal plasticity and protection.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
THE INTERACTION BETWEEN GANGLERON AND HUMAN SERUM ALBUMIN: FLUORESCENCE STUDIES
Ghazaryan A. G., Grigoryan K. R.
Yerevan State University, Department of Chemistry, Yerevan, Armenia [email protected]
The methods of fluorescence spectroscopy combined with UV/vis and other spectroscopic methods are widely used for monitoring molecular interactions involving proteins [1, 2]. We present the results of the studies on Gangleron (spasmolytic and anaesthetizing drug) interaction with human serum albumin (HSA). HSA fluorescence intensities at the quenching were determined taking into account the inner filter effect detected in this system due to fluorescence properties of HSA and Gangleron. Dynamic quenching mechanism of HSA fluorescence was established based on the studies carried out at different temperatures (298, 303 and 309K). This mechanism was confirmed by UV/vis absorption spectroscopy method. Stern-Volmer constant
(KSV), quenching rate constant (kq) and activation energy of bimolecular quenching (Еа) are presented in table.
Table. Stern-Volmer quenching constants and activation energy for the interaction of HSA and Gangleron
K k E T, K SV q a (x103, M-1) (x1011, M-1.s-1) (kJ . mol-1) 298 5.4 5.4 303 7.3 7.3 49.64 309 11.0 11.0
The results have showed that the values of KSV increased with increasing temperature, which indicates that the quenching mechanism of Gangleron-HSA interaction was initiated by dynamic collision. ______
[1] Jing Zhang, Hui-Hui Sun, Ye-Zhong Zhang et al., Interaction of Human Serum Albumin with Indomethacin: Spectroscopic and Molecular Modeling Study, J.Solun Chem., 2012, 4 : 422. [2] K. R. Grigoryan and A. G. Ghazaryan, Characterization of the platinum (II) dimethylsulfoxide complex binding to bovine serum albumin by fluorescence spectroscopy method, Global Journal of Analytical Chem., 2012, 3 : 1.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
ANNEXIN 11 EXPRESSION PATTERN IN SCHIZOPHRENIA
Ghazaryan H.
Institute of Molecular Biology NAS RA, Yerevan, Armenia [email protected]
Schizophrenia is a complex severe psychiatric disorder of polygenic nature. Identification of a whole complex of schizophrenia related candidate genes may in a sufficient degree improve early diagnostics of this disorder. It is known that apoptotic processes may alter the neuronal network and are involved in the pathogenesis of several psychiatric diseases, such as schizophrenia. Annexin 11 (ANXA11) has complex and essential functions in several biological pathways, including apoptosis and proliferation. The aim of this study was to investigate mRNA expression of the ANXA11 gene in schizophrenia patients in comparison with healthy subjects (controls). Total RNA was isolated from peripheral blood of 66 schizophrenia patients and 99 healthy subjects of Armenian population. The mRNA expression was determined by quantitative real-time polymerase chain reaction (RT-PCR) using PSMB2 as housekeeping gene. Data analysis was based on Students’ t-test. The results obtained indicated that ANXA11 mRNA expression was upregulated in peripheral blood of patients in comparison with controls (patients vs. controls, mean±SEM: 0.92±0.16 vs. 0.44±0.09, p=0.0051). In conclusion, our findings suggest that over- expression of the ANXA11 gene is involved in apoptotic alterations in schizophrenia and contribute to pathomechanisms of this disorder. Further investigations are required to extend this observation and find relation to the disease clinical phenotypes.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
XO REGULATES PURINE CATABOLISM BY FEEDBACK MECHANISM
1Gyongyan S.A., 1, 2Manucharyan T. G., 2Danielyan K.E.
1Yerevan State University, Department of Biochemistry, Yerevan, Armenia 2H. Buniatian Institute of Biochemistry NAS RA, Yerevan, Armenia Contact person: [email protected]
There are numerous publications, evidencing about the primer, regulating role of the hypoxanthine/xanthine existence and its catabolism, proving the existence of the feedback mechanism, where the regulative enzyme stands the XOR in the row of the purines metabolic pathway. For instance, Edwards NL et. al have performed the small clinical trial with the infusion of the radiolabeld [8-(14)C] adenine to four patients with gout as well as to the patients suffering from Lesch-Nyhan syndrome. Five days after infusion it became clear that the mean cumulative excretion of radioactivity after adenine administration to patients not receiving and receiving (off and on) allopurinol therapy was 6.1% and 3.6% of infused radioactivity for gouty subjects and 15.9% and 20.8% for the Lesch-Nyhan patients (Edwards, Recker et al. 1981). Bleisch et all have shown that allopurinol, besides inhibiting uric acid synthesis, reduced the rate of degradation of AMP (Bleisch, Sillero et al. 1994). In our current investigation we have analyzed whether the utility of all compounds entering into the purine catabolizing pathway might be regulated by XO. Activity of XO was evaluated based on the method introduced by Litwack et all (Litwack, Bothwell et al. 1953). Protein quantity was calculated based on the Bradford methos. Human brain derived cell culture was grown in accordance of Mattason’s method (Mattson and Ruchlik 1990). We have used Student t-test as well as ONE-WAY-ANOVA. The activity of XO in the rat brain in the presence of different substrates was the following in comparison with the control (1.2104±0.0000, p<0.05): for histidine 2.2190±0.4707, in the presence of allopurinol-1.6138±0.2690, for riboflavin 1.6138±0.1345 (with allopurinol-1.4651±0.8773), for adenosine-1.6138±0.2017 (with allopurinol-1.2776±0.1345), for desoxyadenosine-2.2997±0.5245, (with allopurinol-1.2507±0.0672). XO activity in the human brain derived cell culture was equal for control to 2.4498e-3±1.6352e-4, in the presence of xanthine to 3.3159e-3±4.5807e-4 and with allopurinol to 2.9447e-3±1.0203e-4: for riboflavin 2.2766 e3±3.7474e-4 (with allopurinol 4.0088e-3±3.5803e-4), for adenosine- 3.1675e-3±6.7497e-4 (with allopurinol 2.5736e-3±6.8458e-4), for desoxyadenosine- 3.0190e- 3±7.3017e-4, (with allopurinol -2.3508e-3±1.0995e-3). We concluded that regulation of XO by feedback mechanism might suppress the catabolism of purines and serve as a basis for the initiation of cells proliferative processes.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
XANTHINE OXIDOREDUCTASE IS A KEY ENZYME OF PURINE CATABOLISM REGULATION
1Gyongyan S.A., 1, 2Manucharyan T. G, 2Danielyan K.E, 2Kevorkyan G.A., 2 Chailyan S.G.
1Yerevan State University, Department of Biochemistry, Yerevan, Armenia 2H. Buniatian Institute of Biochemistry NAS RA, Yerevan, Armenia Contact person: [email protected]
Xanthine Oxidase (XO; EC 1.3.22) as well as Xanthine Dehydrogenase (XDH; EC 1.17.1.4) are two enzymes responsible for the last steps of purine metabolism, hydroxylation of a wide variety pyrimidine, pterin, and aldehyde. There are numerous publications evidencing not directly about regulating role of the hypoxanthine/xanthine existence and its catabolism in the row of the purine metabolic pathway. Activities of XO were evaluated based on the method introduced by Litwack et all [1]. Protein quantity was calculated based on the Bradford method. We have used Student t-test as well as ONE-WAY-ANOVA. We have chosen the best condition for evaluation of XO activity in the homogenate. The activity of XO in the rat brain in the presence of different substrates was the following in comparison with the control (1.2104±0.0000, p<0.05): for histidine 2.2190±0.4707, in the presence of allopurinol1.6138±0.2690, for riboflavin 1.6138±0.1345 (with allopurinol- 1.4651±0.8773), for adenosine-1.6138±0.2017 (with allopurinol-1.2776±0.1345), for desoxyadenosine-2.2997±0.5245, (with allopurinol-1.2507±0.0672). We concluded, that the regulation of Xanthineoxidoreductase by feedback mechanism might suppress the catabolism of purines and serve as a basis for the initiation of cells proliferative processes.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
PRODUCTION OF CELLULASE BY THE HALOALKALIPHILIC STRAINS OF STREPTOMYCES ISOLATED FROM SALINE-ALKALINE SOILS OF ARARAT PLAIN, ARMENIA
Hakobyan A., Panosyan H., Trchounian A.
Yerevan State University, Faculty of Biology, Department of Microbiology and Biotechnology of Plants and Microorganisms ,Yerevan, Armenia [email protected]
Cellulase is an industrially important enzyme, which is extensively used in food, textile and paper industry. A potential challenging area, where cellulases would have a central role, is the bioconversion of cellulosic biomass for bioethanol production. The ability to hydrolyze cellulose is widely distributed among many genera of the domain of Bacteria. Representatives of the genus Streptomyces are attractive industrial organisms due to high growth rate, extracellular secretion of cellulases and biosafety capacity. Although the alkaliphilic microorganisms that can produce cellulases have been studied widely, limited reports are available about haloalkaliphilic producers of cellulases. In this study, cellulolytic enzyme activity of 5 moderately haloalkaliphilic Streptomyces strains isolated from saline-alkaline soils of Ararat Plain (Armenia) was examined. Production of cellulolytic enzyme by isolates was detected on carboxymethylcellulose (CMC) containing medium after 4 days of incubation at 37 °C. Two haloalkaliphilic streptomyces strains phenotypically identified as Streptomyces roseosporus A3 and S. griseus A5 were selected as active producers of cellulase. The effect of NaCl concentration on enzyme activity and pH stability was also tested on CMC-agar plate containing from 0 to 25 % NaCl, pH 5-11. The highest crude enzyme activity (3000-3300 U/mg) was observed at pH 8 and 37 °C in a medium that was supplemented with 2% and 5% NaCl, respectively for S. roseosporus A3 and S. griseus A5. Further optimization of the reaction medium condition will provide an opportunity to increase the enzyme production of the isolates.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
CREATINE/CREATINE KINASE SYSTEM IN THE CELLULAR MECHANISMS OF EHRLICH ASCITES CARCINOMA AND THERAPY WITH NON-PATHOGENIC STRAINS OF ESCHERICHIA COLI
Hovhannisyan M.R., Avagyan H.Kh., Movsesyan H.A., Alchujyan N.Kh., Movsesyan N.A.
H.Buniatian Institute of Biochemistry NAS RA,Yerevan, Armenia [email protected]
Currently bacteria have shown promising and significant potency in eradicating established tumors found in pre-clinical mouse tumor models. The creatine/creatine kinase (CK) system plays a key role in cellular energy buffering and transport. The new data suggest the role of the creatine/CK system in cancer cell survival and tumor progression. We studied metabolic pattern of creatine in malignancy and following bacterial therapy. Ehrlich ascites carcinoma (EAC) cells were injected intraperitoneally to develop tumor in two-month-old wild mice and 11 days later, both the intracellular CK activity and the creatine content were analyzed in peritoneal cavity and blood. The CK activity increased in the peritoneal leukocyte 5 times , and in homogenates and cytoplasm 2,9 times, whereas in mitochondria it dropped approximately 5 times compared to control, respectively. Simultaneously, the CK activity increased dramatically 6, 21, and 27 times, in the blood leukocyte homogenates, cytoplasm and mitochondria, respectively, and in plasma 2.4 times. Treatment of EAC-bearing mice by live non-pathogenic clinical strains of Escherichia coli prolonged mice survival up to 75% and inhibited tumor growth with subsequent decrease in the ascites fluid volume. E. coli-therapy completely canceled the activation of the cytosolic CK and restored to control levels the mitochondrial CK activity in peritoneal leukocytes of mice with EAC, as well as decreased of about 5 times the level of creatine in the ascites. Bacterial treatment also tended to normalize the CK activity in blood leucocyte and decreased twice the creatine content in plasma interfering with tumor growth. We have shown for the first time the diverse effects of live non-pathogenic clinical strains of E. coli on the creatine/CK system in ascites and blood that might be involved in the intracellular mechanisms of E. coli-therapy in cancer.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
TO METABOLISM OF TRANSMITTER AMINO ACIDS IN RAT BRAIN
Khachatryan N. Kh., Vardanyan A.G., Kamalyan R.G.
H.Buniatian Institute of Biochemistry NAS RA, Yerevan, Armenia [email protected]
Glutamine is the main source of brain glutamate and GABA and its additional administration in organism corrects balance of these transmitter amino acids with contrast action on the neuronal activity what is important in the brain function disturbance. In the present work it is studied the influence of intraperitoneal glutamine and GABA-T inhibitor ethanolamine-O- sulphate (EOS) administration on the concentration of amino acids and ammonia in rat brain. It was shown that glutamine administration led to insignificant brain GABA level increase without changes of glutamate and ammonia content. EOS administration leads to significant increase of GABA that considerably increases in common glutamine and EOS administration. The lesser output is obtained when asparagine and EOS were administered in spite of glutamine level increasing. However, the study of possible glutamine formation from asparagine in brain homogenates did not give a uniquely answer to the evidence of the similar process. The asparagine incubation with brain homogenates leads to some output of glutamine and bicarbonate amino acids but not GABA. Ketoglutarate stimulates formation of those amino acids and inhibits added glutamate utilization in ATP presence and absence. The analogical canvas is notes in brain mitochondria. Asparagine promotes glutamate utilization and ammonia formation from the latter. ATP and NAD promote glutamate using by mitochondria. The first one increases aspartate, glutamine and ammonia output from endogenous glutamate, while the second one increases only ammonia level. Ketoglutarate moves amino acids metabolism to side of glutamate synthesis. The endogenous and added ketoglutarate is useed intensly in mitochondria, particularly the latter in the presence of ATP and pyridoxalphosphate. However, there are some indications to glutamine synthesis from asparagine. This question demands additional investigations with use of label asparagine.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
NEW METHOD FOR PURIFICATION OF XANTHINE OXIDASE
1, 2Manucharyan T. G., 1Gyongyan S.A., 2Danielyan K.E.
1Department of Biochemistry, Yerevan State University, Armenia 2H. Buniatian Institute of Biochemistry, NAS, Armenia Contact person: [email protected]
McManaman J.L. with colleages have purified from the liver of the rats Xanthine Oxidase (XO; EC 1.3.22) as well as Xanthine Dehydrogenase (XDH; EC 1.17.1.4). During the experiments were used benzamidine-Sepharose based affinity chromatography as well as different types of electrophoreses 1. The results of isoelectric focusing are evidencing about the proteins with different values of pI: 6,13 and 6,23. Also, there was found out one more minor fraction with pI 6,07. However, it is necessarily to mention that all purified proteins were presented as an isoforms of XO. During further presentations authors do not mention about the possible existence of XD isoforms 1. We are presenting here the new method of XO purification with the utility preparative electrophoresis. Activity of XO was evaluated based on the method introduced by Litwack et all2. It was developed the new method based on the existing one 3, for the purification of XO from the liver of rats with the final utility of the preparative electrophoresis with further extraction of the proteins from the gel and evaluation of XO activity. For the calculation of the statistic we have used ONE-WAY-ANOVA (results were considered significant, when p<0.05) There were visualized by phoresis two bands possessing with the XO activity. One of the bands had an approximate molecular weith equal to 300 kDa, the other one had approximately 40 kDa molecular weight. We have delineated also the activity of the XO in the pure fractions. Activity was detected in the first as well as 5th fractions (0.2317±0.0133-control, 0.3008±8.9803e-3-xanthine, 0.2155±0.0135-allopurinol in low concentration, 0.2326±0.0117- allopurinol in high concentration, p<0.01 for first fraction and 0.2004±0.0544, 0.2298±0.0353, 0.2188±0.0393, 0.2249±0.0272 for 5th fraction). We have developed new method of XO purification and found out the XO activity in 2 different fractions with 10 times difference in molecular weights.
References: 1. McManaman, J. L.; Shellman, V.; Wright, R. M.; Repine, J. E., Purification of rat liver xanthine oxidase and xanthine dehydrogenase by affinity chromatography on benzamidine-sepharose. Arch Biochem Biophys 1996 332(1), 135-41. 2. Litwack, G.; Bothwell, J. W.; Williams, J. J.; Elvehjem, C. A., A colorimetric assay for xanthine oxidase in rat liver homogenates. J Biol Chem 1953, 200, (1), 303-310. 3. Engerson, T. D.; McKelvey, T. G.; Rhyne, D. B.; Boggio, E. B.; Snyder, S. J.; Jones, H. P., Conversion of xanthine dehydrogenase to oxidase in ischemic rat tissues. J Clin Invest 1987, 79(6), 1564-70.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
CATALYTIC ACTIVITY OF Mg²⁺- AND Ca²⁺-DEPENDENT ATPases IN MITOCHONDRIAL TISSUES OF SOME SEVAN FISHES
Margaryan A.S., Badalyan R.B., Simonyan A.A.
H.Buniatian Institute of Biochemistry of NAS RA, Yerevan, Republic of Armenia [email protected]
In the presented work activities of Mg²⁺- and Ca²⁺-ATPases in the isolated mitochondria of brain, liver and skeletal muscle of Sevan fishes - crucian, khramulya and barbel were studied. Comparing the obtained data it is possible to make the following conclusions.
The enzyme acivities of Mg²⁺- and Ca²⁺-dependent ATPases in brain, liver and skeletal muscle mitochondria of investigated fishes tissues are higher than general activities of ATPases. Particularly, it was observed in case Mg²⁺-dependent ATPase, in all of investigated tissues mitochondria. Such elevation of the enzyme activity was also observed especially in mitochondria of skeletal muscles. It is obvious that there are different pathways of activation of ATPase fishes tissues. High activities of those ATPase in mitochondria of skeletal muscle were observed. The enzyme acivities of Mg²⁺- and Ca²⁺-dependent ATPases in brain, liver and skeletal muscle mitochondria of Sevan Crucian is higher than in Sevan Khramulya and Sevan Barbel. Taking into account the obtained results, we can conclude that the activities of the enzymes were characterized by tipe and tissue-specifics.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
MEFV GENE EXPRESSION DURING MACROPHAGE ACTIVATION
Nersisyan L.R., Arakelyan A.A.
Institute of Molecular Biology of the NAS RA, Yerevan, Armenia [email protected]
Familial Mediterranean fever (FMF, MIM 249100) is a prototypical recessively inherited autoinflammatory disease most commonly affecting the ethnic groups originating from around the Mediterranean Sea. In 1997, the gene (MEFV) responsible for FMF was identified on the chromosome 16p13.3. To date, more than 100 FMF-associated mutations have been detected and characterized. MEFV has been shown to be expressed in neutrophils, macrophages, and lymphocytes, however, the exact biological functions of pyrin are yet to be elucidated: it is not known what role MEFV plays in immunity, particularly in inflammation. Considering the crucial role of macrophages in development of inflammatory responses during FMF, the aim of this study was to assess MEFV gene expression during classical and alternative macrophage activation. Using publicly available RNA-sequencing data, we evaluated differential expression of MEFV gene using negative binomial distribution model, as well as constructed protein-protein interaction networks of MEFV gene for the cases of classical and alternative activation of macrophages. The results of our study showed that the expression of MEFV is upregulated in alternatively activated macrophages (p < 0.05) compared to classically activated macrophages. Analysis of protein-protein interaction networks has shown that MEFV overexpression may be involved in inhibition of IL-1beta production during alternative activation of macrophages. In conclusion, our data suggest that MEFV gene may have its specific role in activation of macrophages during FMF inflammatory response.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
FTIR STUDY OF THE REDOX PROPERTIES OF CYSTEINE IN THE PRESENCE OF DIMETHYLSULFOXIDE
Papanyan Z., Markarian S.
Yerevan State University, Department of Chemistry, Yerevan, Armenia [email protected]
Cysteine, cystine and methionine are natural sulfur containing amino acids that are important constituents of proteins like ceratins, many enzymes, and are also found in free state. The diverse redox chemistry expressed by these molecules can be explained by the ability of sulfur atom to exist in different oxidation states and take part in many reactions. Another important molecule involved in the biological cycle of sulfur is dimethylsulfoxide (DMSO), which has many technical and biomedical applications and is of significant interest. It is important to investigate possible interactions between DMSO and cysteine and such reports published in the literature by the date are numerous. However there seem to be discrepancies on the exact mechanism of the reaction and forming products. In this work the reaction of L- cysteine with DMSO in aqueous solutions is studied by FTIR spectroscopy under mild conditions. On the basis of obtained IR spectroscopic data it is found that an oxidative conversion of L-cysteine to L-cystine takes place, DMSO is reduced to dimethylsulfide and the water is split off. The reaction can be represented as follows:
A schematic representation of the reaction between cysteine and DMSO: the reagents and the products
Other products, such as cysteic acid, may be obtained under more severe conditions (acidic medium, heating), which agrees with the data presented in literature.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
VOLUMETRIC PROPERTIES OF AOT/N-HEPTANE/DMSO-WATER REVERSE MICELLAR SYSTEMS
Shahinyan G.A., Sargsyan H.R., Markarian S.A.
Yerevan State University, Department of ChemistryYerevan, Armenia [email protected]
Volumetric properties of sodium bis(2-ethylhexyl) sulfosuccinate (AOT)/n-heptane/ dimethylsulfoxide (DMSO) - water reverse micellar systems have been investigated by the method of densitometry. The investigation of such systems is important, because they are considered as a models of membranes due to some structural similarities with biological membranes and the change of the ratio of the components will change the distribution of vitamin E and thus to control its movement inside these systems [1]. The purpose of this work is to find out how the apparent molar volume of polar phase changes depending on the degree of hydration when the water is replaced by the mixed solvent DMSO/water with different volume ratios. By measuring the values of density of systems AOT/heptane and AOT/heptane/water- DMSO in different ratios of mixed solvent the values of apparent molar volume of polar phase have been determined. It can be assumed from the results that the presence and the increase of the concentration of DMSO in the polar phase lead to the growth of density and apparent molar volume of polar phase. It can be explained by that fact, that the increase of concentration of DMSO strengthens the interaction between DMSO and water. In addition the polarity of the polar phase decreases and the vital interaction between molecules of polar phase and AOT decays. Thus, the presence and the increase of the amount of DMSO in the polar phase in AOT/n- heptane/DMSO-water system produce the rise of polar phase apparent molar volume, which is observed by increasing both degree of hydration and temperature.
Referenses:
1 S.A. Markarian, J.D. Grigoryan, H.R. Sargsyan, Int. J. Pharm., 353, 2008, 52–55.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
PRODUCTION OF THERMOSTABLE ALPHA-AMYALSE BY BACILLUS SP. IRANIAN S2 USING SOLID STATE FERMENTATION
Sharifi Alghabpoor S., Panosyan H., Popov Yu., Trchounian A.
Yerevan State University, Faculty of Biology, Department of Microbiology and Biotechnology of Plants and Microorganisms, Yerevan, Armenia [email protected]
Representatives of Bacillus and related genera have been found to be the best candidate for commercial production of thermostable α-amylases. The production of α-amylases has traditionally been carried out using submerged fermentation. Nowadays solid state fermentation system appears as a promising alternative technology because of simple technique, low capital investment, lower levels of catabolite repression and end product inhibition, low waste water output, better product recovery, and high quality production.Application of agro-industrial residues (such as wheat bran) in solid state fermentation bioprocesses solves pollution problems as well. The object of study was bacilli strain Bacillus sp. IranianS2 isolated from geothermal soil samples collected from Gandom-Beryan in Lut desert, Iran. Production of α-amylase under solid-state fermentation by Bacillus sp.IranianS2 has been studied using wheat bran producing waste as substrates.The influences of incubation time, inoculum size, incubation temperature and pH, additional carbon and nitrogen sources on the production of α-amylase were investigated. The highest enzyme production expressed as units per mass of dry substrate (96±3 U/g) was observed after 72 h incubation. The optimum temperature for α-amylase production was observed at 55°C and pH 5.5. Production parameters were optimized as inoculums size 10 % (volume per mass) and substrate:moisture ratio 1:1. Among the defined carbohydrates, the addition of glucose (0.05 g/g dry substrate) has significantly improved the production of α- amylase (128±5 U/g). Supplementation of different nitrogen sources (0.02 g/g) showed decline in enzyme production. Since the enzymes of these strain has a broad pH range of activity, moderate thermostability, and appropriate temperature profile and could be produced on cheap substrates, therefore, it can be suitable candidate to be used as an additive for starch, biofuel and detergent industries.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
EXCITATION - EMISSION MATRIX FLUORESCENCE SPECTROSCOPY STUDIES OF DIMETHYLSULFOXIDE EFFECT ON HUMAN SERUM ALBUMIN STABILITY
Shilajyanand H. A., Grigoryan K. R.
Yerevan State University, Department of Chemistry, Yerevan, Armenia [email protected]
Fluorescence spectroscopy has become a dominating technique in the areas of biochemistry and molecular genetics. When the emission spectrum is obtained for all excitation wavelengths of a sample a data matrix of fluorescence intensities is combined as en excitation emission spectrum. We present the results of excitation-emission matrix (EEM) fluorescence studies of dimethylsulfoxide (DMSO) effect on human serum albumin (HSA). EEM fluorescence spectra of HSA are measured at the presence of DMSO at 250C. Results are presented in the table.
Table: 3D fluorescence spectral characteristics of HSA, HSA-DMSO systems
System HSA HSA - 5% DMSO HSA - 20% DMSO peak I peak II peak I peak II peak I peak II t, min F, a.u. 0 336.7 326.0 161.1 396.3 - 371.3 90 329.0 328.2 161.5 390.6 - 368.7 270 329.0 328.2 98.1 336.9 - 308.0
Two typical fluorescence peaks (peak 1 and peak 2) can be observed in 3D fluorescence spectrum: peak 1(λex / λem = 230/338 nm/nm) and peak 2 (λex / λem = 280/344 nm/nm). Peak 1 reveals fluorescence spectral behavior of Trp and Tyr residue and peak 2 may mainly represent the fluorescence characteristics of the polypeptide backbone structures [1, 2]. Fluorescence characteristics of peak 1 and peak 2 demonstrate that the presence of DMSO affects not only on the polarity of the microenvironment of Trp and Tyr residues but at higher concentrations it causes structural changes in polypeptide backbone structure of protein as well. ______
[1] F.L. Cui, J. L. Wang, Y. R. Cui, J. P. Li., Fluorescent investigation of the interactions between N-(p- chlorophenyl)-N′-(1-naphthyl) thiourea and serum albumin: Synchronous fluorescence determination of serum albumin. Anal. Chim. Acta, 2006, V. 571, P. 175-183. [2] К.Р. Григорян, А.А. Шиладжян, Молекулярные взаимодействия при низких температурах в систах САЧ- диэтилсульфоксид(дипропилсульфоксид)-вода по данным тушения фляоресценции, Журнал физической химнн, 2013, T. 87, №5, C. 800-803.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
ALTERATIONS OF LIPID MODIFICATION PROCESSES IN THE PERIPHERAL BLOOD MONONUCLEAR CELLS IN MALIGNANCY
Torgomyan T.R., Lazyan M.P., Hakobyan G.V., Batikyan T.B., Ghazaryan R.A., 1Alexanyan K.A., 1Galstyan H.M., Tadevosyan Y.V.
Laboratory of Regulation of Cellular Activity, Institute of Molecular Biology, NAS RA, Yerevan, Armenia 1 National Center of Oncology after V.Fanarjyan, MH RA, Yerevan, Armenia [email protected]
The involvement of cell plasma membrane (PM) lipids in the regulatory mechanisms of various important PM-associated processes is well documented. These compounds by quick and reversible changes in their composition and structure (microdomain) organization respond rapidly to different environmental perturbations, especially leading to the pathologies. The present study aimed to clarify the regularities of membrane neutral lipids (NL) early (5 sec) and long-term (60 min) acylation by exogenous [14C]arachidonic acid (AA) in human peripheral blood crude mononuclear cells (MNC) at norm and in ovarian (OC) or breast (BC) cancers. The data obtained indicate that in MNC mechanisms of NLs fatty acid content quick and sustained modification by exogenous AA were significantly altered in OC and BC compared to norm and were also distinctly individual for each patient. It’s important that disturbances revealed in MNC obtained from patients with OC and BC were identical with those observed earlier in 3 different forms of leukemia. Thus, we conclude that the regular alterations revealed are common for diverse forms of malignancy studied, and can be used as additional testing parameters for cancer definition, assessment of the depth of pathological state as well as for discovery of new personalized modes for cancer treatment.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
OXIDATIVE STRESS AND PATHOMECHANISMS OF ISCHEMIC STROKE
Tsakanova G.V., Ayvazyan V.A., Boyajyan A.S.
Institute of Molecular Biology NAS RA, Yerevan, Armenia
Oxidative stress (OS) is a key pathogenic factor leading to uncontrolled cell damage and death, which badly influence ischemic stroke (IS) progression and outcome. Molecular mechanisms involved in development of these processes are not clear yet, which limits the identification of therapeutic targets for IS. The majority of data in this field were obtained in animal models of stroke, which not adequately reflect the pathogenesis of stroke in humans. Important indicators of development of OS are elevated levels of lipid-, protein-, and DNA- derived oxidized products and reduced antioxidant capacity of organism. The aim of current study is to reveal molecular mechanisms responsible for development of OS in human IS on systemic level. To achieve this goal, by using blood serum samples of IS-affected and healthy subjects, we performed: 1) assessment of peculiarities in development of systemic OS in IS by measuring blood levels of oxidized derivatives of lipids, proteins and DNA, including lipid hydroperoxides, lipofuscin, 3-nitrotyrosine, 8-isoprostaglandine-F2, matrix metalloproteinases- 9, and 8-hydroxi-2’-desoxiguanosine; (2) evaluation of the functional state of antioxidant system in IS at the systemic level by determining the total capacity of low-molecular non-enzymatic water-soluble antioxidants (TAC), ferroxidase activity of ceruloplasmin (FAC) and the activities of enzymes superoxide dismutase and catalase in the blood. Series of methods, including photochemiluminescent and colorimetric assays, fluorescent analysis, and enzyme-linked immunosorbent assay were performed. The results obtained demonstrated that IS is characterized by dysfunction of adaptive reactions of organism, which is reflected by unremarkable increase in TAC and FAC in response to OS at day 1 of IS-onset. Furthermore, it was shown that the action of systemic OS in IS at the 1-st day of IS onset appears on the level of oxidative damage of lipids. Damaging effects of OS in IS on the first day of stroke onset are manifested by lipid oxidative modification reactions, and by dysfunction of free-radical scavenger system.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
STUDY AND ASSESSMENT OF MICROBIAL COMMUNITIES IN NATURAL AND COMMERCIAL BIOLEACHING PROCESSES
1Vardanyan A.K., 2Khachatryan A.N., 3Stepanyan S.Kh.
Institute of Microbiology, SPC “Armbiotechnology”, NAS RA , Yerevan, Armenia [email protected], 2 [email protected], 3 [email protected]
The progress in biohydrometallurgy requires multilateral study of communities of chemolithotrophic bacteria in natural ecosystems characterized with extremely low pH values and high concentration of metals, elucidating physiological peculiarities of dominating species as well as the evaluation of their potential for the application in metal recovery. The exceptional diversity of ecogeographical conditions of Armenia and the richness of non-ferrous and rare metals represent a great and valuable potential for investigation of biodiversity of acidophilic chemolithotrophic sulphur and iron oxidizing bacteria as well as for isolation of new highly active strains and characterization of their microbial communities for application in biohydrometallurgical processes in order to inhance their efficiency. The study of biodiversity of chemolithotrophic bacteria has both commercial and environmental importance. Studies of physiological and biotechnological properties of these relevant strains from the bioleaching communities will allow to achieve comprehensive insight of interspecies relationships as a substantial prerequisite for enhancement of the efficiency of metal bioleaching. The determination and description of sulphur and iron oxidizing bacteria included in the bioleaching processes are very essential for performing their monitoring in natural ecosystems, as well as for the improvement of existing technologies of metal recovery and development of new technologies.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
FRET MICROSCOPY FOR REAL-TIME MONITORING OF cGMP INDUCED BY PRP-1
Yeranosyan L.A.
H. Buniatian Institute of Biochemistry NAS RA, Yerevan, Yerevan, Armenia [email protected]
Discovery of cardioactive neurohormones (produced by the NSO and NPV) and atrium natriuretic factors from the atria [2], points to the existence of the functional regulatory system referred to as “neuroendocrine hypothalamus— endocrine heart” [1].Now we hypothesize that this system plays an exclusively important role in the general adaptation of the organism, and especially under cardiac stress. Of current interest is to identify functionally relevant biochemical mechanisms of interconnection between cardioactive signalling molecules from the hypothalamus (proline rich peptide-1 – PRP‐1) and cardiac second-messenger cascades particularly the cGMP dependent protein kinase signal transduction pathway. PRP-1 (discovered by Prof. A. Galoyan and coworkers) [3] is currently under investigation as drug candidates for the treatment of myocardial infarction and other cardiac diseases. To address this issue we have studied the effect of PRP -1 on cGMP in ventricular cardiomyocytes since cGMP- dependent pathwayis a common target in the combined pharmacological treatment of heart failure (cGMP-elevating drugs). We employed living cells isolated from transgenic cGMP– sensor expressing mice used as a model for real-time FRET imaging. Recently constructed cGMP sensor named red cGES-DE5 was used in our experiments. In this construct cGMP binding domain from a phosphodiesterase (PDE5) was sandwiched between T-Sapphire and dimer2 [4]. Upon addition of cGMP, red cGES-DE5 exhibited a decrease in FRET. FRET acceptor / donor fluorescence emission ratio (FL590/515) was measured in the cells stimulated with a different concentration of PRP-1, IBMX (non-specific PDE inhibitor ) and CNP (increase cGMP production through guanylylcyclase-B). The FRET signals at the cardiomyocytes, as indicated by the ratio of GFP to RFPfluorescence for red cGES-DE5, showed that bath application of PRP-1 (1µM) induced an insignificant elevation of the cGMP level.
Referenses: 1. GaloyanAA(2010), “Concepts of Neuroendocrine Cardiology and Neuroendocrine Immunology, Chemistry and Biology of Signal Molecules”; Neurochem Res 35:2001–2017. 2. Cole BR, Currie MG, Geller DM, Michener ML, Saper CB,Schwartz D, Standaert DG (1985) Atriopeptins as cardiac hormones hypertension 7(4):469–482 3. Galoyan AA (2004) Brain Neurochemistry Cytokines: Immune Response and Neuronal Survival. Kluwer Academic/Plenum Publishers, New York 4. NikolaevVO, Gambaryan S and Lohse MJ(2006) Fluorescent sensors for rapid monitoring of 5. intracellular cGMP. Nature Methods , 3 (1)
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New Aspects in Molecular Biotechnology and Biochemistry 2013
NEUROTROPHIN FAMILY GENE AS POTENTIAL TARGET FOR SCHIZOPHRENIA
Zakharyan R. V., Boyajyan A. S.
Institute of Molecular Biology NAS RA, Yerevan, Armenia [email protected]
Alterations in neurodevelopment are thought to contribute to the etiology of schizophrenia (SCZ), a complex mental disorder. Genes encoding synaptic plasticity regulatory proteins might be considered as candidate genes for this disorder. In order to clarify the role of netrin G1 (NTNG1) and brain derived neurotrophic factor (BDNF) proteins in SCZ their genetic variants and blood levels in disease-affected and healthy subjects were studied. Genotyping for NTNG1 gene rs628117 and BDNF rs6265 polymorphisms were performed using PCR-SSP and blood plasma levels were assessed by ELISA. The NTNG1 rs628117 genotypes were equally distributed in the groups whereas the carriers of minor rs6265*A allele of the BDNF gene were overrepresented among SCZ patients with compared to controls (pcorrected=0.006). Furthermore, the AA genotype correlated with the earlier onset of the disease (p=0.024). Also, we found decreased BDNF plasma levels both in treated and non- treated patients compared to controls. Comparative analysis of BDNF blood levels regarding rs6265 genotypes indicated that, compared to individuals homozygous for the standard rs6265*A carriers had decreased BDNF levels. Thus, the pathogenesis of SCZ is characterized by genetically predetermined decreased blood BDNF levels and the rs6265*A minor allele can be considered as a risk factor for SCZ in Armenian population.
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New Aspects in Molecular Biotechnology and Biochemistry 2013
AB INITIO AND DFT THEORETICAL STUDY OF THE INTERACTION OF L-AA/DESO
Zatikyan A.L., Markarian S.A.
Yerevan State University, Department of Chemistry, Yerevan, Armenia [email protected]
To determine the structure, charges and energies of stable conformers of various types L- AA/DESO complexes in gas phase and solution, Ab initio Hartree-Fock (HF) and DFT methods were used with the GAUSSIAN 03 software package. The optimized geometric parameters and interaction energies for various complexes at different theories have been estimated. The self- consistent reaction field (SCRF) was used to calculate the effect of DESO as a solvent on the geometry, energy, dipole enhancement and vibrational frequencies of interacting complexes. The obtained data on the basis of Raman and FT IR studies of L-AA/sulfoxide mixtures show that very strong interactions take place between L-AA and DESO [1]. The solvent effect has been studied using the Onsager models. The results indicate that the polarity of the solvent has played an important role on the structures and relative stabilities of different complexes. The results obtained show that there is a satisfactory correlation between experimental and theoretical predictions, that is to say, for L- AA/DESO systems in gas phase all three types of the complexes are possible, while in condensed phase only one type of complex is predominate: complex with double-hydrogen bonded structure (fig.).
[1] A.L. Zatikyan, E.A. Kazoyan, S. Bonora, S.A. Markaryan. J. Appl. Spectrosc. 2008, 75, 664-668
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New Aspects in Molecular Biotechnology and Biochemistry 2013
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