Medical Genetic Research Center

Moscow, Russia

Medical Genetic Research Center

Moscow, Russia
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Dadkhah E.,Mashhad University of Medical Sciences | Dadkhah E.,George Mason University | Naseh H.,Omid Hospital | Farshchian M.,Mashhad University of Medical Sciences | And 10 more authors.
Archives of Iranian Medicine | Year: 2013

Background: Esophageal squamous cell carcinoma (ESCC) is the second-most frequently diagnosed cancer in Northeast Iran, often diagnosed in advanced stages. No standard early diagnostic guideline has been proposed to date and current therapeutic modalities are not effective. Detection of tumor-specific biomarkers, which is the goal of this study, could prove useful in the diagnosis of ESCC. Methods: To better understand the gene expression profile of ESCC, we analyzed tumor samples and corresponding adjacent normal tissue from ESCC patients by Chemiluminescent Human Cancer GEArrays. Candidate genes were verified by real-time PCR. Results: Out of 440 cancer-related genes included in the array, 71 were overexpressed compared to normal tissue, with significant differences in 11 genes. There were 108 genes underexpressed, with significant differences in 5 genes. Until now, the AP2M1, FTL, UBE2L6, HLA-C, and HSPA8 overexpressed genes and XRCC5, TP53I3 and RAP1A underexpressed genes were not reported in ESCC. We chose the MMP2, HLA-G, and XRCC5 markers from 58 Iranian ESCC patients to verify the expression validity by real-time PCR. The microarray results were confirmed with two-tailed significance levels of P = 0.003 (MMP2), P = 0.000 (HLA-G) and P = 0.002(XRCC5). Analysis performed for the candidate genes using GNCpro online software highlighted two pathways, an immuno-modulatory response and DNA replication and repair. We successfully performed and validated Chemiluminescent GEArray gene expression profiling in ESCC. Several biomarkers that might be related to tumorigenesis in ESCC were identified. Conclusion: Immuno-modulatory and DNA repair pathways could be used as targets to locate specific diagnostic, prognostic, and therapeutic biomarkers for ESCC.


PubMed | Russian National Research Medical University, Research Institute of Human Morphology and Medical Genetic Research Center
Type: Journal Article | Journal: Bulletin of experimental biology and medicine | Year: 2016

Short-term cell culturing on basement membrane matrix is a common and very convenient in vitro model of angiogenesis. We studied the possibility of interaction of multipotent stromal cells from the umbilical cord and Ea.hy926 endothelial cells on this model at the early and late periods of the experiment. Multipotent stromal cells alone and in combination with endothelial cells formed an unstable tubular network. Clusters formed after its disassembling later became the sprouting centers in co-culture of the two cell types, but not in pure culture of multipotent stromal cells. Multipotent stromal cells with CD31+ phenotype constitute the structural basis of newly formed stable 3D capillary-like network. Prolongation of the time of culturing and combination of the two in vitro models of angiogenesis (tubulogenesis and sprouting) allowed more complete assessment of the angiogenic potential of multipotent stromal cells.


Kostyuk S.V.,Medical Genetic Research Center | Smirnova T.D.,Medical Genetic Research Center | Efremova L.V.,Medical Genetic Research Center | Konkova M.S.,Medical Genetic Research Center | And 3 more authors.
Bulletin of Experimental Biology and Medicine | Year: 2010

NO synthesis by endothelial cells plays an important role in normal function of the cardiovascular system. This work was designed to evaluate the role of variations in properties of extracellular DNA in the regulation of NO synthesis. We studied the effect of four DNA samples with various base sequences (50 ng/ml) on functional activity of endothelial cells HUVEC during 24-h culturing. Human DNA fragments with high content of CG repeats increased intracellular content of NO and its metabolites (nitrites and nitrates) and accelerated oxidation of nitrites to nitrates. Changes in the content of NO metabolites after 24-h culturing was shown to depend on the expression of gene for inducible, but not for endothelial NO synthase. Increased expression of inducible NO synthase positively correlated with an increase in the content of mRNA for the adapter protein MyD88, but did not depend on TLR9 gene expression that encodes protein receptor for CG-DNA recognition. The intracellular concentration of MyD88 mRNA did not depend on the content of TLR9 mRNA. The existence of a variety of DNA-binding receptors apart from TLR9 receptor on the surface of endothelial cells was hypothesized. Activation of these receptors by extracellular DNA fragments stimulates expression of the adapter protein MyD88. © 2010 Springer Science+Business Media, Inc.


Stepanova A.A.,Medical Genetic Research Center | Krasovsky S.A.,Research Institute of Pulmonology | Polyakov A.V.,Medical Genetic Research Center
Russian Journal of Genetics | Year: 2016

A study of Russian cystic fibrosis (CF) patient DNA was conducted to assess the incidence frequency of 19 mutations, namely CFTRdele2,3(21kb), F508del, I507del, 1677delTA, 2143delT, 2184insA, 394delTT, 3821delT, L138ins, 604insA, 3944delGT, G542X, W1282X, N1303K, R334W, and 3849 + 10kbC > T, S1196X, 621 + 1g > t, and E92K of the CFTR gene. We also sought to determine the estimated CF frequency in Russian Federation. In addition, we determined the total information content of the approach for 19 common mutations registration in the CFTR gene, 84.6%, and the allelic frequencies of the examined mutations: three mutations were observed with a frequency exceeding 5% (F508del, 53.98%, E92K, 6.47%, CFTRdele2,3(21kb), 5.35%); other mutations were observed with frequencies ranging from 0.13 to 3.0%. The CF population carrier frequency was 1 in 38 subjects, while the predicted CF frequency was 1 in 5776 newborns. © 2016, Pleiades Publishing, Inc.


Baskova I.P.,Moscow State University | Alekseeva A.Yu.,Moscow State University | Kostyuk S.V.,Medical Genetic Research Center | Neverova M.E.,Medical Genetic Research Center | And 2 more authors.
Biomeditsinskaya Khimiya | Year: 2012

The medicinal leech salivary cell secretion (SCS) may stimulate NO-production in cultures of human endothelium cells (HUVEC) and rat cardiomiocytes (RCM). This effect was detected using a NO specific reagent, - the complex Cu2+ with a fluorescein derivative (Cu-F1). NO had also been detected in the cells by fluorescent electronic microscopy and determined quantitatively in the cells and in culture fluid by the fluorescence method. SCS stimulated NO synthesis in HUVEC cells (but not in RCM) is accompanied by NO release into intercellular space. Localization of NO synthesis centers is presented and it is shown that the increase in NO levels during the SCS action on HUVEC and RCM is associated with the increase in the activity of eNOS/nNOS, but not iNOS. In endothelial cells SCS activates nitrosylation processes, assessed by the increase of nitrite-ions in the culture medium. It is therefore important to use Cu-F1, other than Griss-reagent, during the first hour of analysis of NO synthesis. The NO-depended mechanism of SCS action on endothelial cells might be a factor in providing of its positive action in hirudotheraphy.


Nakhaei Moghaddam M.,Islamic Azad University at Mashhad | Hashemi Beidokhti M.,Islamic Azad University at Science and Research of Fars | Amel Jamehdar S.,Mashhad University of Medical Sciences | Ghahraman M.,Medical Genetic Research Center
Iranian Journal of Basic Medical Sciences | Year: 2014

Objective(s): blaCTX-M and blaPER are two genes that encode class A extended-spectrum β-lactamases (ESBLs) and can be responsible for therapeutic problems. This study was carried out to evaluate the molecular properties of these genes in clinical isolates of Enterobacteriaceae by polymerase chain reaction (PCR), restriction digestion and sequencing. Materials and Methods: During six months, starting from January 2012, one hundred clinical isolates of Enterobacteriaceae were collected from urinary samples. The ESBL-producing isolates were detected by phenotypic confirmation test. After plasmid extraction, blaPER and blaCTX-M genes were detected using PCR by specific primers. The blaCTX-M PCR products were digested with Taq1, and two of the blaCTX-M genes were sequenced. Results: Phenotypic tests showed that 27 (27%) isolates were ESBL producers with the highest frequency for Klebsiella pneumoniae (47.4%) and Escherichia coli (17.9%). Twenty six (26%) of Enterobacteriaceae isolates harbored the blaCTX-M gene, and none of them had blaPER. The restriction analysis of PCR products showed that all blaCTX-M amplified products had the same patterns. Both sequenced bacteria were CTX-M-15 type ESBL carriers. Conclusion: The results of this study showed the blaCTX-M-15 gene in Enterobacteriaceae isolates for the first time in Mashhad, Iran. High degrees of associated resistance to co-trimoxazole and gentamicin were found in ESBL producers. Therefore, an integrated and regular management of antibiotic prescription need to be trained in our society.


Baskova I.P.,Moscow State University | Alekseeva A.Y.,Moscow State University | Kostyuk S.V.,Medical Genetic Research Center | Neverova M.E.,Medical Genetic Research Center | And 2 more authors.
Biochemistry (Moscow) Supplement Series B: Biomedical Chemistry | Year: 2013

Using a novel NO-specific reagent, the complex of Cu2+ with a fluorescein derivative (Cu-FL), stimulation of NO production by the medicinal leech salivary cell secretion (SCS) has been demonstrated for the first time in cultures of human endothelial cells (HUVEC) and rat cardiomyocytes (RCM). NO was detected in the cells by fluorescent electronic microscopy and determined quantitatively in the cells and in the culture fluid by the fluorescence method. SCSstimulated NO synthesis in HUVEC but not in RCM cells was accompanied by NO release into the intercellular space thus determining its subsequent distribution. Localization of intracellular NO synthesis centers is presented and it is shown that the increase in NO levels during the SCS action on HUVEC and RCM is associated with the increase in the activity of eNOS/nNOS, but not iNOS. In the endothelial cells SCS-activated nitrosylation processes, estimated by the increase of nitrite-ion content in the culture medium. It is therefore important to use Cu-FL, rather than Griss-reagent, during the first hour of analysis of NO synthesis. It is possible that the NO-depended mechanism of the SCS action on endothelial cells may be a factor responsible for the positive effect of SCS during hirudotheraphy. © 2013 Pleiades Publishing, Ltd.


PubMed | Islamic Azad University at Science and Research of Fars, Mashhad University of Medical Sciences, Medical Genetic Research Center and Islamic Azad University at Mashhad
Type: Journal Article | Journal: Iranian journal of basic medical sciences | Year: 2014

blaCTX-M and blaPER are two genes that encode class A extended-spectrum -lactamases (ESBLs) and can be responsible for therapeutic problems. This study was carried out to evaluate the molecular properties of these genes in clinical isolates of Enterobacteriaceae by polymerase chain reaction (PCR), restriction digestion and sequencing.During six months, starting from January 2012, one hundred clinical isolates of Enterobacteriaceae were collected from urinary samples. The ESBL-producing isolates were detected by phenotypic confirmation test. After plasmid extraction, blaPER and blaCTX-M genes were detected using PCR by specific primers. The blaCTX-M PCR products were digested with Taq1, and two of the blaCTX-M genes were sequenced.Phenotypic tests showed that 27 (27%) isolates were ESBL producers with the highest frequency for Klebsiella pneumoniae (47.4%) and Escherichia coli (17.9%). Twenty six (26%) of Enterobacteriaceae isolates harbored the blaCTX-M gene, and none of them had blaPER . The restriction analysis of PCR products showed that all blaCTX-M amplified products had the same patterns. Both sequenced bacteria were CTX-M-15 type ESBL carriers.The results of this study showed the blaCTX-M-15 gene in Enterobacteriaceae isolates for the first time in Mashhad, Iran. High degrees of associated resistance to co-trimoxazole and gentamicin were found in ESBL producers. Therefore, an integrated and regular management of antibiotic prescription need to be trained in our society.

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