1/2017

Author

page: - pp
DOI: 10.11134/btp.1.2017.1
Aitkulova A.M.1, Zholdybayeva E.V.2, Shashkin C.S.3, Amanbayeva D.3, Aitasheva Z.G.1
1Al-Farabi Kazakh National University, 
71, al-Farabi ave., Almaty, 050040, Kazakhstan
2National Center for Biotechnology, 
13/5, Korgalzhyn road, Astana,010000, Kazakhstan
3National Neurosurgeon Center, 
34/1, Turan ave., Astana,010000, Kazakhstan

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Abstract

Parkinson’s disease (PD) was first described as a shaking palsy syndrome by James Parkinson in 1817. PD is a chronic progressive disease found in all populations of the world and is the second most common neurodegenerative disease. The major pathological feature of PD is progressive degeneration of the nigrostriatal system, which leads to the loss of dopaminergic neurons in the substantia nigra pars compacta. The degeneration of the nigrostriatal system and subsequent loss of striatal dopamine contribute to the cardinal clinical motor symptoms of PD: tremor, rigidity, bradykinesia, and postural instability. The most common symptoms of PD only become apparent when 50–80% of dopaminergic neurons of the substantia nigra have already degenerated. The mechanism underlying the loss of dopaminergic neurons during PD development remains unclear. Currently, there are no simple and reliable diagnostic tests for PD. Current research efforts are increasingly focused on elucidating the molecular basis of the disease. Studies have revealed the relationship between PD and environmental risk factors and genetic predisposition. Despite the identification of genes and loci involved in PD development, the molecular mechanisms that promote disease onset and progression are not fully understood.

Keywords: Parkinson’s disease, dopaminergic neurons, neurodegeneration, genes

 

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Author

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DOI: 10.11134/btp.1.2017.2
Mektepbayeva D.1, Janzakova D.2, Shaimerdenova M.1, Karapina O.3, Mu X.4, Kenneth A.5, Akilbekova D.1
1National Laboratory Astana, Nazarbayev University,
KabanbayBatyr Ave, 53, Astana, 010000, Kazakhstan
2Nazarbayev University,
KabanbayBatyr Ave, 53, Astana, 010000, Kazakhstan
3Nazarbayev University Research and Innovation System, 
KabanbayBatyr Ave, 53, Astana, 010000, Kazakhstan
4University of Texas Health Science Center,
3SCR6 #3706 1881 East Road, Houston, TX 77054, USA
5Alibek Kenneth, Locus Solutions, LLC, 30500 Aurora Rd Ste 180 Solon, 
OH 44139-2776, USA

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Abstract

Recent studies have demonstrated that common factors present in aggressive cancer cells and embryonic progenitors influence cancer aggressiveness. This review will summarize the cancer-related properties of various embryonic microenvironments. The anticancer effects of embryonic microenvironments on various types of cancer cells and tumors are associated with epigenetic alterations and down regulation of tumor-suppressor genes and the Nodal signaling pathway. The ability to reprogram the tumorigenic phenotype of cancer cells depends on the stage and type of embryonic microenvironment, and reprogramming capacity significantly decreases after organogenesis. Understanding the mechanisms underlying the molecular reprogramming of cancer cells in various embryonic models will help identify potential targets for cancer therapy. 
Keywords: Cancer, cell reprogramming, epigenetics, embryonic microenvironments, differentiation

 

 

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Author

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DOI: 10.11134/btp.1.2017.3
Sikhayeva N.S.1,2,Dzharmukhanov Zh.M.1, Nakysh A.T.3, Smagulova S.K.4, Shaimerdenov S.A.4, Baidurin S.A.3, Zholdybayeva E.V.1, Ramankulov E.M.1,2
1National center for biotechnology, 
Korgaldzyn av., 13/5, Astana, 010000, Kazakhstan
2L.N. Gumilyov Eurasian National University, 
Munaitpasova str., 5, Astana, 010000, Kazakhstan
3Astana Medical University, 
Koshkarbayeva str., 66, Astana, 010000, Kazakhstan
4City polyclinic №4,
Shevchenko str., 4, Astana, 010000, Kazakhstan

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Abstract

Diabetes and obesity are multifactorial diseases that are major global health problems. The pathogenic mechanisms of these diseases include several pathways operating under the influence of the interaction of genetic and environmental factors. According to numerous studies, there are multiple loci that contribute to the risk of multifactorial (polygenic) diseases. It was previously demonstrated that polymorphisms in the GCK, YKT6and INSIG2 genes are associated with the risk of type 2 diabetes and obesity in different populations. This study aimed to investigate the association of GCK, YKT6 and INSIG2 allelic polymorphisms and risk of type 2 diabetes and obesity in a Kazakh population. The study included patients diagnosed with type 2 diabetes. The control group consisted of healthy volunteers who were invited to participate in the study during a routine health examination. A total of 662 participants of Kazakh nationality were genotyped for two single nucleotide polymorphisms: rs4607517 and rs7566605. The results of genetic analysis revealed a statistically significant association between polymorphisms inGCK, YKT6 and INSIG2 and risk of type 2 diabetes (rs4607517: OR = 0.70, CI = 0.51–0.95, p = 0.024 in the additive model) and obesity (rs4607517: OR = 0.20, CI = 0.03–0.82, p = 0.045; rs7566605: OR = 1.82, CI = 1.11–3.02, p = 0.018 in the recessive model). In addition, these polymorphisms were associated with different blood parameters, including hemoglobin, erythrocytes, and mean platelet volume. In conclusion, we observed a statistically significant association between GCK, YKT6 and INSIG2 polymorphisms and risk of developing type 2 diabetes and obesity in a Kazakh population.

Keywords: Single nucleotide polymorphism, type 2 diabetes, obesity, Kazakh population

 

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Author

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DOI: 10.11134/btp.1.2017.4
Bokebayev Zh.T.1, Kukiev I.S.1, Krivoruchko T.N.2, Sergazy Sh.D.2, Shulgau Z.T.2, Tolmacheva O.V.2, Abuova G.T.3, Zhaugasheva S.K.4, Gulyayev A.E.2
1AO «Medical University Astana»,
Beibitshilik Ave., 49A, Astana, 010000, Kazakhstan
2RSE«National Center for Biotechnology»,
Kurgalzhinskoe highway, 13/5, Astana, 010000, Kazakhstan
3South-Kazakhstan State Pharmaceutical Academy,
Al-Farabi, 1, Shymkent, 160019, Kazakhstan
4Karagandy State Medical University,Gogolya str., 40, Karaganda, 100000, Kazakhstan
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Abstract

The aim of the study was to compare the antimicrobial, anti-inflammatory and wound healing effects of an angiogenin gel with those of a modified version of the gel containing the antimicrobial drug dioxidine. Modifications of the angiogenin gel by the addition of dioxidine resulted in enhanced antimicrobial action against the main pathogens of wound purulent-septic infections: Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, and Escherichia coli. The presence of dioxidine in the angiogenin gel did not modify the anti-inflammatory effect, which was similar to that of the standard angiogenin gel. The wound healing effect of the angiogenin gel containing dioxidine was enhanced in a burn wound model, as accelerated epithelization was observed, potentially due to the contribution of the antimicrobial component to the healing process.

Keywords: Angiogenin, dioxidine, antimicrobial, anti-inflammatory, wound healing

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Author

page: - pp
DOI: 10.11134/btp.1.2017.5
IssabekovaA.S.1, ZhunusovaM.S.2, RamanculovE.M.1, KulyyassovA.T.1
1Republican State Enterprise «National Center for Biotechnology» under the Science Committee of Ministry of Education and Science of the Republic of Kazakhstan,
Astana, 010000, 13/5, Kurgalzhynskoye road,
2L.N.Gumilyov Eurasian National University,
5, Munaitpasov str.,010000, Astana, Kazakhstan
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Abstract

The expression of transgenes and control of gene expression in mammalian cells is an active area of biomedical research. Therefore, the improvement of methods for effective gene transfer into eukaryotic cells is of great interest and remains are search priority. We compared different methods of transient transfection of HEK293T cells, namely calcium phosphate with different pH values of the HBS buffer solution,6.90, 6.95, 7.00, 7.05 and 7.10, as well ascommercial sets of FuGENE and Lipofectamine. Results were analyzed using fluorescence microscopy with green fluorescent protein GFP as a reporter gene. We determined that the optimal pH of the HBS phosphate buffer for transient transfection was 6.95. A small pH deviation in the range of 6.90–7.10 did not significantly modify the expression efficiency of a protein of interest. Among the methods studied, Lipofectamine showed the highest level of transfection efficiency based on expression of the GFP protein, but exceeding the recommended dose of this reagent exerted a toxic effect on the viability of HEK293T cells.

Keywords: Plasmid, DNA, transient transfection, HEK293T

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Author

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DOI: 10.11134/btp.1.2017.6
Li P., Amenov A., Kalendar R., Abeldenov S., Khassenov B.
RSE «National Center for Biotechnology» under the Science Committee, Ministry of Education and Science of the Republic of Kazakhstan,
Astana, Kazakhstan
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Abstract

The invention of loop-mediated isothermal amplification (LAMP) opened new research avenues in the field in diagnostics. LAMP reactions have important advantages as a diagnostic tool.  These advantages include a constant reaction temperature that only requires a simple thermostat instead of a thermocycler and a greater reaction speed from 15 to 30 minutes. The method is based on a specific thermostable polymerase with helicase activity and a set of different primers. We obtained a large fragment of recombinant polymerase I from Geobacillus stearothermophilus expressed in Escherichia coli. Original strain (ATCC 12980) cells were cultivated in nutrient broth to extract genomic DNA and amplify the target gene. After its cloning and expression, the polymerase was purified in an amount of 1.5 mg from 1 L of induction culture of recombinant cells. The purified polymerase was tested and displayed polymerase and helicase activities with no exonuclease activity. These activities were demonstrated in successful LAMP reactions with the amplification of transgenic elements of genetically modified Arabidopsis thaliana using a LAMP primer set for detection of nopaline synthase terminator (T-nos) and another set for detection of the Cauliflower Mosaic Virus 35S promoter. We showed that LAMP reaction products could be detected using an agarose gel and fluorescent dyes.

Keywords: LAMP, Bst, isothermal amplification, Geobacillus stearothermophilus, recombinant protein, diagnostics

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Author

page: - pp
DOI: 10.11134/btp.1.2017.7
DzhakashevaМ.А.1, Lieberzeit Р.А.2
1Ministry of education and science Republic of Kazakhstan,
M. Auezov South-Kazakhstan state university, Taukekhanavenue, 5, Shymkent, 160000, Kazakhstan
2University of Vienna, Univeristatsring 1, 1010 Vienna, Austria
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Abstract

We studied an immobilized biocatalyst process based on a multi-enzymic complex consisting of pectinase, cellulase, hemicellulase and β-glucanase, in whicha polyvinyl alcohol cryogel was used as a carrier. The polyvinyl alcohol cryogel is a viscoelastic nonshattering material, which is unabraded, has favorable operational properties for a long duration, is able to take any granule shape, and is appropriate for different reactors with different operating modes. To form cross-linked enzymatic aggregates, a multi-enzymic complex was precipitated by isopropyl alcohol with the simultaneous addition of a glutaric aldehyde solution as the linking agent and reduced cured grape polyuria as the water-retaining agent and filler. The process was studied to include amulti-enzymic complex to the polyvinyl alcohol cryogel matrix. The reduced grape polyuria stabilized the catalytic exoenzyme activity retention at their cross-linking by glutaric aldehyde and negatively influenced enzymes activity in the complex cross-linked enzymatic aggregates content. The gained high-active biocatalyst immobilized to the polyvinyl alcohol cryogel displayed pH stability and high catalytic stability for a long time without any disturbance of the operating regime, and the process has multiple potential applications in winemaking. Due to aporous structure, the matrix of this carrier did not modify reactions catalyzed by the enzyme and resulted in a quality finished product (ruddy wine).

Keywords: Enzyme immobilization, cross-linked aggregates, pectolytic enzymes, multi-enzymic complex, polyvinyl alcohol

 

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Author

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DOI: 10.11134/btp.1.2017.8
Yagofarova A.Y., Kurmanbaev A.A., Berdimuratov K.T., Ahmetollaev I.A.
Republican State Enterprise «National Center for Biotechnology» under the Science Committee of Ministry of Education and Science of the Republic of Kazakhstan,
13/5, Kurgalzhynskoye road, Astana, 010000, Kazakhstan
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Abstract

We investigated the simultaneous production of electrical power and wastewater sludge recycling by microbial fuel cells (MFC). The constructed MFC were comprised of two chambers, each with an approximate volume of 100ml, which were connected by salt bridge or proton exchange membrane (PEM). The graphitic anode was placed in one chamber, which  was filled with wastewater sludge from a sewage treatment plant of Astana or a medium with acetate and electrogenic strain; a graphite cathode was placed in the other chamber and filled with an electrolyte. Several prototypes were created and tested in our laboratory. The most important parameters such as voltage and amperage were measured during 9 days; the maximum values obtained were 0.6V and 0.7mA. Moreover, the power generated by the MFC made in our laboratory was 0.42mW. Glass wool was selected as the material for PEM. The electrogenic properties of the selected bacterial strains BacillusamylofaciensU15, EnterobacterPs7, and LactobacillusfermentumTB4 were studied. EnterobacterPs7 was more active than the others. The fuel cell blocks were designed to accommodate a voltage increase. Thus, the eight MFC were connected successively. The maximum voltage was 2.6V and amperage 30μA. The maximum power of the electric current was 78.0 μW.

Keywords: Microbial fuel cells, recycling of organic waste, electricity, electrogenic bacteria

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Author

page: - pp
DOI: 10.11134/btp.1.2017.9
Chervyakova O.V., Strochkov V.M., Tailakova E.T., Sultankulova K.T., Samdybayev N.T.
pgt. Gvardeiskiy, Korday District, Zhambyl region, 080409, Kazakhstan
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Abstract

The ability of capripoxviruses to elicit cellular and humoral immune responses, incorporate extended DNA sequences into host genomes, and display a safety profile are important considerations in the development of vector vaccines for animals and humans. The aim of our study was to design and construct recombinant plasmid DNAs for the recombination of capripoxvirus genomes. Two genetic constructs were developed, pINT-TKsppv and pINT-RRsppv, which contained sequences of sheeppox virus genome DNA insufficientfor virus replication (thymidine kinase gene and small ribonucleotidereductase gene) as well as a cassette for target gene expression and selective marker (gpt). Control of the expression of target genes will be driven by a synthetic early/late vaccinia virus promoter. These constructs will be used in the generation of recombinant sheeppox viruses expressing target antigens.

Keywords: recombinant plasmids, sheeppox virus, directional mutagenesis, dominant selective marker

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