new aspects in molecular biotechnology and biochemistryaab.sci.am/documents/theses.pdf ·...

New Aspects in Molecular Biotechnology and Biochemistry 2013

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Young Scientists' Conference

NNEEWW AASSPPEECCTTSS

IINN MMOOLLEECCUULLAARR BBIIOOTTEECCHHNNOOLLOOGGYY

AANNDD BBIIOOCCHHEEMMIISSTTRRYY

27-28 June, 2013

H. Buniatian Institute of Biochemistry NAS RA

Yerevan, Republic of Armenia

ABSTRACTS


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Contents:

RGANIZING COMMITTEE

5

PREFACE

6

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.

,

Davtyan T.K.

7

GENDER DIFFERENCES OF PARP-1 ACTIVITY IN LIVER

Asatryan A.L., Artsruni I.G., Matinyan K.S., Margaryan A. V., Gevorgyan E.S.

8

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.

9

STUDY OF MUSHROOMS` INTRACELLULAR EXTRACTS ANTI-

INFLAMMATORY AND ANTICANCER ACTIVITY

Avagyan I.A. , Minasbekyan L.A., Nanagulyan S.G.

10

CHANGES IN APOPTOTIC RATE AND SYNAPTIC PLASTICITY IN PATIENTS

WITH POSTTRAUMATIC STRESS DISORDER

Avetyan D.

11

INVESTIGATION OF THE ANTIBACTERIAL ACTIVITY OF PROLINE-RICH

POLYPEPTIDES AGAINST BACILLUS ANTHRACIS

Badalyan A.M., Badalyan Kh. V.

12

REGENERATION OF BURN INJURY UNDER THE INFLUENCE OF OINTMENTS

FROM THE EXTRACTS OF MULBERRY LEAVES AND SILVER BERRY SEEDS

Balasanyan M.G., Soghbatyan L.T., Grigoryan D.S.

13

SOME PROPERTIES OF ANTIBACTERIAL COMPONENT OF LACTIC ACID

BACTERIA ISOLATED FROM ARMENIAN DAIRY PRODUCTS

Bazukyan I., Babayan A., Trchounian A.

14

DO WE HAVE A CHANCE TO TEST ADEQUATELY NEWLY DEVELOPED

DRUGS IN ANIMAL MODELS OF FOCAL ISCHEMIA?

Danielyan K.E.

15

NEW METHOD FOR PURIFICATION OF XANTHINE DEHYDROGENASE

Feresheryan K, Manucharyan T. G, Gyongyan S.A., Danielyan K.E.

16


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QUANTUM CHEMICAL STUDY OF DIMETHYL-AND DIETHYLSULFONES

Gabrielyan L.S., Mkhitaryan A.S., Markarian S.A.

17

ELECTROPHYSIOLOGICAL STUDIES OF LATERAL VESTIBULAR NUCLEUS

AFTER NUCLEOTIDE THERAPY OF DAMAGED SCIATIC NERVE

Gevorgyan L.R., Chavushyan V.A., Simonyan K.V.

18

THE INTERACTION BETWEEN GANGLERON AND HUMAN SERUM ALBUMIN:

FLUORESCENCE STUDIES

Ghazaryan A. G., K. R. Grigoryan

19

ANNEXIN 11 EXPRESSION PATTERN IN SCHIZOPHRENIA

Ghazaryan H.

20

XO REGULATES PURINE CATABOLISM BY FEEDBACK MECHANISM

Gyongyan S.A., Manucharyan T. G, Danielyan K.E.

21

XANTHINE OXIDOREDUCTASE IS A KEY ENZYME OF PURINE CATABOLISM

REGULATION

Gyongyan S.A., Manucharyan T. G, Danielyan K.E, Kevorkyan G.A., Chailyan S.G.

22

PRODUCTION OF CELLULASE BY THE HALOALKALIPHILIC STRAINS OF

STREPTOMYCES ISOLATED FROM SALINE-ALKALINE SOILS OF ARARAT

PLAIN, ARMENIA

Hakobyan A., Panosyan H., Trchounian A.

23

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.

24

TO METABOLISM OF TRANSMITTER AMINO ACIDS IN RAT BRAIN

Khachatryan N. Kh., Vardanyan A.G., Kamalyan R.G.

25

NEW METHOD FOR PURIFICATION OF XANTHINE OXIDASE

Manucharyan T. G., Gyongyan S.A., Danielyan K.E.

26

CATALYTIC ACTIVITY OF Mg²⁺- AND Ca²⁺-DEPENDENT ATPases IN

MITOCHONDRIAL TISSUES OF SOME SEVAN FISHES

Margaryan A.S., Badalyan R.B., Simonyan A.A.

27

MEFV GENE EXPRESSION DURING MACROPHAGE ACTIVATION

Nersisyan L.R., Arakelyan A.A.

28


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FTIR STUDY OF THE REDOX PROPERTIES OF CYSTEINE IN THE PRESENCE OF

DIMETHYLSULFOXIDE

Papanyan Z., Markarian S.

29

VOLUMETRIC PROPERTIES OF AOT/N-HEPTANE/DMSO-WATER REVERSE

MICELLAR SYSTEMS

Shahinyan G.A., Sargsyan H.R., Markarian S.A.

30

PRODUCTION OF THERMOSTABLE ALPHA-AMYALSE BY BACILLUS SP.

IRANIAN S2 USING SOLID STATE FERMENTATION

Sharifi Alghabpoor S., Panosyan H., Popov Yu., Trchounian A.

31

EXCITATION - EMISSION MATRIX FLUORESCENCE SPECTROSCOPY STUDIES

OF DIMETHYLSULFOXIDE EFFECT ON HUMAN SERUM ALBUMIN STABILITY

Shilajyanand H. A., Grigoryan K. R.

32

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.,

Alexanyan K.A., Galstyan H.M., Tadevosyan Y.V.

33

OXIDATIVE STRESS AND PATHOMECHANISMS OF ISCHEMIC STROKE

Tsakanova G.V., Ayvazyan V.A., Boyajyan A.S.

34

STUDY AND ASSESSMENT OF MICROBIAL COMMUNITIES IN NATURAL AND

COMMERCIAL BIOLEACHING PROCESSES

Vardanyan A.K., Khachatryan A.N., Stepanyan S.Kh.

35

FRET MICROSCOPY FOR REAL-TIME MONITORING OF cGMP INDUCED BY

PRP-1

Yeranosyan L.A.

36

NEUROTROPHIN FAMILY GENE AS POTENTIAL TARGET

FOR SCHIZOPHRENIA

Zakharyan R. V., Boyajyan A. S.

37

AB INITIO AND DFT THEORETICAL STUDY OF THE INTERACTION OF L-

AA/DESO

Zatikyan A.L., Markarian S.A.

38

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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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.

http://aab.sci.am/

http://aab.sci.am/

http://aab.sci.am/

http://aab.sci.am/

http://aab.sci.am/

http://aab.sci.am/

http://www.molbiol.sci.am/

http://aab.sci.am/

http://aab.sci.am/

http://www.ysmu.am/

http://aab.sci.am/

http://rau.am/faculties.php?l=0&f_id=2

http://www.ysu.am/site/index.php?lang=1&page=3&p_num=&id=29

http://aab.sci.am/




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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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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.

mailto:[email protected]


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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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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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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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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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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 i n v i 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 w h i t e mi ce . All the mice were infected i.p. with anthrax strain

N55 vaccine (1x107/m ice ) . Th e co n t ro l gr o up r ece iv ed v eh ic l e ( 0 . 4 m l o f 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 th e r a t e o f 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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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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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.



- 15

-

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.



- 16

-

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.



- 17

-

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).



- 18

-

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

[email protected]

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.



- 19

-

THE INTERACTION BETWEEN GANGLERON AND HUMAN SERUM

ALBUMIN: FLUORESCENCE STUDIES

Ghazaryan A. G., Grigoryan K. R.


[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

T, K KSV

(x103, M

-1)

kq

(x1011

, M-1.

s-1

)

Ea

(kJ . mol

-1)

298 5.4 5.4

49.64 303 7.3 7.3

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.



- 20

-

ANNEXIN 11 EXPRESSION PATTERN IN SCHIZOPHRENIA

Ghazaryan H.


[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.



- 21

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XO REGULATES PURINE CATABOLISM BY FEEDBACK MECHANISM

1Gyongyan S.A.,


2Danielyan K.E.

1Yerevan State University, Department of Biochemistry, Yerevan, Armenia

2H. Buniatian Institute of Biochemistry NAS RA, Yerevan, Armenia


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.



- 22

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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


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.



- 23

-

PRODUCTION OF CELLULASE BY THE HALOALKALIPHILIC

STRAINS OF STREPTOMYCES ISOLATED FROM SALINE-ALKALINE

SOILS OF ARARAT PLAIN, ARMENIA

Hakobyan A., Panosyan H., Trchounian A.


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.



- 24

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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.



- 25

-

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.



- 26

-

NEW METHOD FOR PURIFICATION OF XANTHINE OXIDASE


1Gyongyan S.A.,

2Danielyan K.E.

1Department of Biochemistry, Yerevan State University, Armenia

2H. Buniatian Institute of Biochemistry, NAS, Armenia


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.



- 27

-

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.



- 28

-

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.



- 29

-

FTIR STUDY OF THE REDOX PROPERTIES OF CYSTEINE IN THE

PRESENCE OF DIMETHYLSULFOXIDE

Papanyan Z., Markarian S.


[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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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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PRODUCTION OF THERMOSTABLE ALPHA-AMYALSE BY BACILLUS

SP. IRANIAN S2 USING SOLID STATE FERMENTATION

Sharifi Alghabpoor S., Panosyan H., Popov Yu., Trchounian A.


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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EXCITATION - EMISSION MATRIX FLUORESCENCE

SPECTROSCOPY STUDIES OF DIMETHYLSULFOXIDE EFFECT ON

HUMAN SERUM ALBUMIN STABILITY

Shilajyanand H. A., Grigoryan K. R.


[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

t, min peak I peak II peak I peak II peak I peak II

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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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 [14

C]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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OXIDATIVE STRESS AND PATHOMECHANISMS OF ISCHEMIC

STROKE

Tsakanova G.V., Ayvazyan V.A., Boyajyan A.S.


[email protected]

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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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.

http://www.sci.am/resorgs.php?oid=43&langid=1





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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)


http://europepmc.org/abstract/MED/20603173/?whatizit_url_gene_protein=http://www.uniprot.org/uniprot/?query=cyclase&sort=score


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NEUROTROPHIN FAMILY GENE AS POTENTIAL TARGET

FOR SCHIZOPHRENIA

Zakharyan R. V., Boyajyan A. S.


[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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AB INITIO AND DFT THEORETICAL STUDY OF THE

INTERACTION OF L-AA/DESO

Zatikyan A.L., Markarian S.A.


[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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