Draft Genome Sequence of Lactobacillus salivarius KZ-NCB, Isolated from Chicken Cecum

Here, we report the draft genome sequence of Lactobacillus salivarius strain KZ-NCB, which was isolated from the cecum of a healthy chicken from a poultry farm in Kazakhstan.Peer reviewe

MACROPOROUS CELL-LADEN GELATIN/HYALURONIC ACID/CHONDROITIN SULFATE CRYOGELS FOR ENGINEERED TISSUE CONSTRUCTS

Cryogels are a unique macroporous material for tissue engineering. In this work, we study the effect of hyaluronic acid on the physicochemical properties of cryogel as well as on the proliferation of a 3D model of mesenchymal stem cells. The functional groups of the synthesized cryogels were identified using Fourier transform infrared spectroscopy. With an increase in the content of hyaluronic acid in the composition of the cryogel, an increase in porosity, gel content and swelling behavior was observed. As the hyaluronic acid content increased, the average pore size increased and more open pores were formed. Degradation studies have shown that all cryogels were resistant to PBS solution for 8 weeks. Cytotoxicity assays demonstrated no toxic effect on viability of rat adipose-derived mesenchymal stem cells (ADMSCs) cultured on cryogels. ADMSC spheroids were proliferated on scaffolds and showed the ability of the cryogels to orient cell differentiation into chondrogenic lineage even in the absence of inductive agents. Thus, our results demonstrate an effective resemblance to extracellular matrix structures specific to cartilage-like microenvironments by cryogels and their further perspective application as potential biomaterials

Genetic diversity of Francisella tularensis subsp. holarctica in Kazakhstan

Tularemia is a highly dangerous zoonotic infection due to the bacteria Francisella tularensis. Low genetic diversity promoted the use of polymorphic tandem repeats (MLVA) as first-line assay for genetic description. Whole genome sequencing (WGS) is becoming increasingly accessible, opening the perspective of a time when WGS might become the universal genotyping assay. The main goal of this study was to describe F. tularensis strains circulating in Kazakhstan based on WGS data and develop a MLVA assay compatible with in vitro and in silico analysis. In vitro MLVA genotyping and WGS were performed for the vaccine strain and for 38 strains isolated in Kazakhstan from natural water bodies, ticks, rodents, carnivores, and from one migratory bird, an Isabellina wheatear captured in a rodent burrow. The two genotyping approaches were congruent and allowed to attribute all strains to two F. tularensis holarctica lineages, B.4 and B.12. The seven tandem repeats polymorphic in the investigated strain collection could be typed in a single multiplex PCR assay. Identical MLVA genotypes were produced by in vitro and in silico analysis, demonstrating full compatibility between the two approaches. The strains from Kazakhstan were compared to all publicly available WGS data of worldwide origin by whole genome SNP (wgSNP) analysis. Genotypes differing at a single SNP position were collected within a time interval of more than fifty years, from locations separated from each other by more than one thousand kilometers, supporting a role for migratory birds in the worldwide spread of the bacteria.Peer reviewe

GENETIC POLYMORPHISM OF 27 Y-STR LOCI IN THEWESTERN KAZAKH TRIBES FROM KAZAKHSTAN AND KARAKALPAKSTAN, UZBEKISTAN

Data on the genetic polymorphism of 27 Y-STR in Kazakhs of the Junior Zhuz has been presented and analyzed in relation to forensic features. A total of 464 representatives of the Western Kazakh tribes of Kazakhstan (Western Kazakhs, n = 405) and Uzbekistan (Karakalpakstan Kazakhs, n = 59) were examined by the Yfiler Plus set. The data are available in the YHRD under accession numbers YA006010 and YA006009. Genetic analysis (AMOVA and MDS) did not show significant differences between the two groups (Kazakhstan and Karakalpakstan Kazakhs) in terms of Y-chromosome diversity. Both groups are characterized by haplogroup C2a1a2 as a founder effect, which dominated two of the three tribes: Alimuly (67%), Baiuly (74.6%), and Zhetiru (25.8%). At the same time, the phylogenetic network for each tribe found its own clusters within C2a1a2. Western Kazakhs and Karakalpakstan Kazakhs present high values of unique haplotypes (84.44% and 96.61%), discrimination capacity (90.37% and 98.30%), and haplotype diversity (0.9991 and 0.9994). A set of 27 Y-STR loci distinguishes closely related individuals within the Western Kazakh tribes quite well. It is suitable for forensic application, and is also optimal for population genetics studies

Heparin-Conjugated Fibrin Hydrogel with Chondroinductive Growth Factors and Human Synovium-Derived Mesenchymal Stem Cells for the Treatment of Articular Cartilage Defects: Evaluation of Clinical Safety

The purpose of this study was to evaluate the safety of an injectable heparin-conjugated fibrin (HCF) hydrogel containing human synovium-derived mesenchymal stem cells (SDMSCs), TGF-β1, and BMP-4 after implantation into articular cartilage defect in patients with osteoarthritis (OA). The study included 15 OA patients with a mean age of 44.2±18.0 years. The median articular cartilage defect size was 4.9±2.0 cm. HCF hydrogel, containing SDMSCs and growth factors (TGF-β1 and BMP-4), was implanted into the cartilage defect using DUPLOJECT two-syringe device connected with the DUPLOTIP dual lumen cannula. Clinical and radiological outcomes were evaluated with VAS, WOMAC, KOOS, and MOCART. The clinical study results showed that implantation of HCF hydrogel with autologous SDMSCs, TGF-β1, and BMP-4 appeared to be safe and did not show severe adverse events in OA patients. The assessment of clinical outcomes after 6 months showed improvement in VAS, WOMAC, and KOOS scores in all patients. The MOCART evaluation demonstrated an enhancement of cartilage tissue repair in 73.3% of OA patients at 6 months after surgery. Thus, implantation of HCF hydrogel with SDMSCs, TGF-β1, and BMP-4 was safe and demonstrated signs of improvement in articular cartilage repair. The evaluation of the long-term safety and therapeutic efficacy of HCF hydrogel is required in a further clinical study using a larger number of OA patients

Generation of Peptides for Highly Efficient Proximity Utilizing Site-Specific Biotinylation in Cells

Protein tags are peptide sequences genetically embedded into a recombinant protein for various purposes, such as affinity purification, Western blotting, and immunofluorescence. Another recent application of peptide tags is in vivo labeling and analysis of protein–protein interactions (PPI) by proteomics methods. One of the common workflows involves site-specific in vivo biotinylation of an AviTag-fused protein in the presence of the biotin ligase BirA. However, due to the rapid kinetics of labeling, this tag is not ideal for analysis of PPI. Here we describe the rationale, design, and protocol for the new biotin acceptor peptides BAP1070 and BAP1108 using modular assembling of biotin acceptor fragments, DNA sequencing, transient expression of proteins in cells, and Western blotting methods. These tags were used in the Proximity Utilizing Biotinylation (PUB) method, which is based on coexpression of BAP-X and BirA-Y in mammalian cells, where X or Y are candidate interacting proteins of interest. By changing the sequence of these peptides, a low level of background biotinylation is achieved, which occurs due to random collisions of proteins in cells. Over 100 plasmid constructs, containing genes of transcription factors, histones, gene repressors, and other nuclear proteins were obtained during implementation of projects related to this method

Comparative Characterization and Identification of Poly-3-hydroxybutyrate Producing Bacteria with Subsequent Optimization of Polymer Yield

In this work, the strains Bacillus megaterium RAZ 3, Azotobacter chrocococcum Az 3, Bacillus araybhattay RA 5 were used as an effective producer of poly-3-hydroxybutyrate P(3HB). The purpose of the study was to isolate and obtain an effective producer of P(3HB) isolated from regional chestnut soils of northern Kazakhstan. This study demonstrates the possibility of combining the protective system of cells to physical stress as a way to optimize the synthesis of PHA by strains. Molecular identification of strains and amplification of the phbC gene, transmission electron microscope (TEM), extracted and dried PHB were subjected to Fourier infrared transmission spectroscopy (FTIR). The melting point of the isolated P(3HB) was determined. The optimal concentration of bean broth for the synthesis of P(3HB) for the modified type of Bacillus megaterium RAZ 3 was 20 g/L, at which the dry weight of cells was 25.7 g/L−1 and P(3HB) yield of 13.83 g/L−1, while the percentage yield of P(3HB) was 53.75%. The FTIR spectra of the extracted polymer showed noticeable peaks at long wavelengths. Based on a proof of concept, this study demonstrates encouraging results

Draft Genome Sequences of Three Pasteurella multocida Strains Isolated from Domestic Animals in Kazakhstan

We report here the draft genome sequences of three strains of Pasteurella multocida isolated in Kazakhstan from domestic animals that died due to hemorrhagic septicemia

CYTOPROTECTIVE ACTIVITY OF NEWLY SYNTHESIZED 3-(ARYLMETHYLAMINO)-6-METHYL-4-PHENYLPYRIDIN-2(1H)-ONES DERIVATIVES

Currently, studies are being conducted on the possible role of the cytoprotective effect of biologically active substances in conditions of cerebral hypoxia or cardiomyopathies. At the same time, oxidative stress is considered one of the important mechanisms of cellular cytotoxicity and a target for the action of cytoprotectors. The aim of this study is to search for derivatives of 3-(arylmethylamino)-6-methyl-4-phenylpyridin-2(1H)-ones. The probability of cytoprotective action was assessed by measuring cell viability using two tests (with neutral red dye and MTT test). It was found that some derivatives of 3-(arylmethylamino)-6-methyl-4-phenylpyridin-2(1H)-ones under the conditions of our experiment had a pronounced cytoprotective activity, providing better cell survival in vitro, including the MTT test and conditions of blood hyperviscosity. To correlate the obtained results in vitro, molecular docking of the synthesized derivatives was also carried out. The standard drug omeprazole (co-crystallized with the enzyme) was used as a standard. It was shown that all synthesized derivatives of 3-(arylmethylamino)-6-methyl-4-phenylpyridin-2(1H)-ones had higher affinity for the selected protein than the standard gastro-cytoprotector omeprazole. The studied derivatives of 3-(arylmethylamino)-6-methyl-4-phenylpyridin-2(1H)-ones also fully satisfy Lipinski’s rule of five (RO5), which increases their chances for possible use as orally active drugs with good absorption ability and moderate lipophilicity. Thus, the results obtained make it possible to evaluate derivatives of 3-(arylmethylamino)-6-methyl-4-phenylpyridin-2(1H)-ones as having a relatively high cytoprotective potential

Draft Genome Sequence of Moraxella bovoculi Strain KZ-1, Isolated from Cattle in North Kazakhstan

Place: Washington Publisher: Amer Soc Microbiology WOS:000572063900018Moraxella bovoculi strain KZ-1 was isolated from cattle that had symptoms of infectious bovine keratoconjunctivitis (IBK) in northern Kazakhstan. Here, we report the draft genome sequence of this strain