Clinical and Genetic Characteristics of Preeclampsia.

Preeclampsia (PE) is a severe complication of pregnancy accompanied by arterial hypertension, edema, or proteinuria with impaired functioning of various organs and systems. It is also an important medical and social problem, which has been one of the leading causes of maternal and perinatal mortality and morbidity worldwide. Despite the achievements of modern medicine, the etiology of this pathology is still unknown. Recently, many scientists have especially focused on the study of genetic factors underlying the etiopathogenesis of PE, namely, the contribution of individual polymorphic loci of various candidate genes. The current study aimed to investigate the clinical characteristics of PE and the contribution of the polymorphic loci rs1042838 of Progesterone Receptor (PGR) gene and rs8068318 of the T-Box Transcription Factor 2 (TBX2) gene to the development of PE. The study was conducted on 219 women with PE with the mean±SD age of 26.52±5.51 years and 329 women with the physiological course of pregnancy as the control group with the mean±SD age of 26.27±4.88 years. In total, 64.20%, 68.29%, 16.44%, 98.63%, and 35.48% of women with PE had increased systolic and normal diastolic blood pressure (SBP and DBP) values, proteinuria, edema, and overweight (BMI≥25), respectively. In the control group, 100%, 1.53%, 1.12%, and 35.48% of cases had normal SBP values with no proteinuria, DBP>90 mm Hg, edema, and overweight (BMI≥25), respectively. An association was observed between the CC genotype of the rs8068318 polymorphism of the TBX2 gene with the risk of developing PE in women with PE (OR=2.12, 95%CI: 1.14-3.92, P=0.02). In addition, there was an association between the rs8068318 TBX2 polymorphic locus with lower SBP (Me=140, Q25 - Q75 130 - 142.5, P=0.01) and PBP (Me=50, Q25 - Q75 40 - 55, P<0.01). According to the GeneCards database, the TBX2 gene, a member of a phylogenetically conserved gene family, is located on the long arm of chromosome 17 and encodes the TBX2 T-box transcription factor protein, which is a regulator of the transcriptional activity of various genes (i.e., it suppresses the expression of CDKN2A (p19/ARF), inhibits cyclin-dependent kinase p21 Cip1 (CDKN1A), and affects the expression of MYC, RAS, BRCA1, and BRCA2 genes).

Risk Factors and Methods for Predicting Ovarian Hyperstimulation Syndrome (OHSS) in the in vitro Fertilization.

Ovarian hyperstimulation syndrome (OHSS) is the most severe and life-threatening complication of assisted reproductive technologies (ART). OHSS is based on an excessive ovarian response to ovarian stimulation; however, the pathogenesis has not been fully understood yet. The most serious complications of OHSS are thromboembolic complications and ovarian torsion. The current study describes the risk factors for the development of ovarian hyperstimulation syndrome and proposes a method for specific prediction of this syndrome. This study was designed to investigate 671 therapeutic cycles in the IVF program during 2009-2018. All patients were divided into two groups. Group one (n=56) included women who developed OHSS during the IVF procedure. Group two (n=615) consisted of women who did not have this complication during the IVF procedure. All the observation and examination outcomes were entered into a specially developed questionnaire, and then into a Microsoft Excel spreadsheet. The data were processed by variable statistics using Statistica 10.0. Analyzing of the recorded data revealed that the rate of OHSS was higher in the group of younger women, aged 30.76±3.67 years, in comparison with those aged 32.78±4.40 years in the group of patients without OHSS (p<0.05). The analysis of the initial phase of the reproductive system has confirmed that the group of patients with OHSS had a higher level of prolactin, 462.84±191.56 mIU/L in comparison with 363.43±187.84 mIU/L, which corresponded to the group of women without OHSS (p<0.05). In our results, 7.15±1.04% of cases with OHSS had obesity, while of the patients from the group without OHSS suffered from it (p<0.05). OHSS is the most severe iatrogenic complication of ART, therefore it is extremely important to consider its risk factors and take timely preventive measures. This study has established a high relationship between the studied risk factors and ovarian hyperstimulation syndrome and proposed a model for predicting this syndrome.

[Induction of apoptotic endonuclease EndoG with DNA-damaging agents initiates alternative splicing of telomerase catalytic subunit hTERT and inhibition of telomerase activity hTERT in human CD4+ and CD8+ T-lymphocytes]. / Induktsiia apoptoticheskoi éndonukleazy EndoG povrezhdaiushchimi DNK agentami vyzyvaet al'ternativnyi splaising kataliticheskoi sub"edinitsy telomerazy hTERT i ingibirovanie aktivnosti telomerazy v CD4+ I CD8+ T-limfotsitakh cheloveka.

Activity of telomerase catalytic subunit hTERT (human Telomerase Reverse Transcriptase) can be regulated by alternative splicing of its mRNA. At present time exact mechanism of hTERT splicing is not fully understood. Apoptotic endonuclease EndoG is known to participate this process. EndoG expression is induced by DNA damages. The aim of this work was to investigate the ability of DNA-damaging agents with different mechanism of action to induce EndoG expression and inhibit telomerase activity due to the activation of hTERT alternative splicing in normal activated human CD4+ and CD8+ T-lymphocytes. All investigated DNA-damaging agents were able to induce EndoG expression. Cisplatin, a therapeutic compound, producing DNA cross-links induced the highest level of DNA damages and EndoG expression. Incubation of CD4+ and CD8+ T-cells with cisplatin caused the changes in proportion of hTERT splice variants and inhibition of telomerase activity.

Intracellular Localization of Apoptotic Endonuclease EndoG and Splice-Variants of Telomerase Catalytic Subunit hTERT.

The activity of telomerase catalytic subunit hTERT (human telomerase reverse transcriptase) can be regulated by alternative splicing of its mRNA. The mechanism of hTERT splicing is not understood in detail. Apoptotic endonuclease EndoG is known to participate in this process. In the present work, the intracellular colocalization and mRNA levels of EndoG and hTERT splice-variants in normal and apoptotic cancer cells were studied. We found that the development of apoptosis increased the expression of EndoG and changed the ratio of hTERT splice-variants, which decreased the telomerase activity in the cells. The development of apoptosis was accompanied by changes in the amount of mRNA and in the localization of EndoG and hTERT splice-variants in the cytoplasm, nuclei, and mitochondria of the cells. The suppression of EndoG expression using RNA interference prevented induction of the α+ß- splice-variant of hTERT and inhibition of the telomerase activity. A high degree of the intracellular colocalization of EndoG and hTERT was shown. The changes in the expression and localization of EndoG corresponded with changes in the amount and localization of hTERT splice-variants. These data confirm the participation of EndoG in the alternative splicing of mRNA of the telomerase catalytic subunit and in regulation of the telomerase activity.

[Cisplatin-induced apoptotic endonuclease EndoG inhibits telomerase activity and causes malignant transformation of human CD4+ T lymphocytes]. / Indutsirovannaia tsisplatinom ékspressiia apoptoticheskoi éndonukleazy EndoG vyzyvaet ingibirovanie aktivnosti telomerazy i zlokachestvennuiu transformatsiiu CD4+ T-limfotsitov cheloveka.

Alternative splicing of telomerase catalytic subunit hTERT pre-mRNA (human Telomerase Reverse Transcriptase) regulates telomerase activity. Increased expression of non-active splice variant hTERT results in inhibition of telomerase. Apoptotic endonuclease EndoG is known to participate in hTERT alternative splicing. Expression of EndoG can be induced in response to DNA damages. The aim of this study was to determine the ability of a DNA-damaging compound, cisplatin, to induce EndoG and its influence on alternative splicing of hTERT and telomerase activity in human CD4+ Т lymphocytes. Overexpression of EndoG in CD4+ T cells downregulated the expression of active full-length hTERT variant and upregulated its non-active spliced variant. Reduction of full-length hTERT caused downregulation of telomerase activity, shortening of telomeres length during cell divisions, converting cells to the replicative senescence state, activation of apoptosis and finally cell death. Few cells survived and underwent malignant transformation. Transformed cells have increased telomerase activity and proliferative potential compare to initial CD4+ T cells. These cells have phenotype of T lymphoblastic leukemic cells and are able to form tumors and cause death in experimental mice.

Apoptotic Endonuclease EndoG Inhibits Telomerase Activity and Induces Malignant Transformation of Human CD4+ T Cells.

Telomerase activity is regulated by an alternative splicing of mRNA of the telomerase catalytic subunit hTERT (human telomerase reverse transcriptase). Increased expression of the inactive spliced hTERT results in inhibition of telomerase activity. Little is known about the mechanism of hTERT mRNA alternative splicing. This study was aimed at determining the effect of an apoptotic endonuclease G (EndoG) on alternative splicing of hTERT and telomerase activity in CD4+ human T lymphocytes. Overexpression of EndoG in CD4+ T cells downregulated the expression of the active full-length hTERT variant and upregulated the inactive alternatively spliced variant. Reduction of full-length hTERT levels caused downregulation of the telomerase activity, critical telomere shortening during cell division that converted cells into the replicative senescence state, activation of apoptosis, and finally cell death. Some cells survive and undergo a malignant transformation. Transformed cells feature increased telomerase activity and proliferative potential compared to the original CD4+ T cells. These cells have phenotype of T lymphoblastic leukemia cells and can form tumors and cause death in experimental mice.

The Lactobacillus rhamnosus and Lactobacillus fermentum strains from human biotopes characterized with MLST and toxin-antitoxin gene polymorphism.

The diversity of Lb. rhamnosus and Lb. fermentum strains isolated from feces, saliva, and the vaginal cavity of 18-22-year-old healthy women residing in central regions of the Russian Federation has been characterized. The results obtained using multilocus sequence typing were identical to those obtained with the analysis of genetic and genomic polymorphism in TA systems. Different as well as identical Lb. rhamnosus and Lb. fermentum sequence types (ST) were isolated from various parts of the body of the same person. Identical ST were also isolated from different women, suggesting that such strains belong to a common pool of strains circulating among the population members. Our results demonstrate that TAs are suitable for characterizing intra-specific diversity of Lb. rhamnosus and Lb. fermentum strains. The advantage of using polymorphisms in TA systems for genotyping is based on the weak number of genes used, and consequently, less time is required for the analysis.

[Apoptotic endonuclease EndoG regulates alternative splicing of human telomerase catalytic subunit hTERT]. / Reguliatsiia al'ternativnogo splaisinga kataliticheskoi sub"edinitsy telomerazy hTERT apoptoticheskoi éndonukleazoi EndoG.

Human telomerase catalytic subunit hTERT is subjected to alternative splicing results in loss of its function and leads to decrease of telomerase activity. However, very little is known about the mechanism of hTERT pre-mRNA alternative splicing. Apoptotic endonuclease EndoG is known to participate this process. The aim of this study was to determine the role of EndoG in regulation of hTERT alternative splicing. Increased expression of b-deletion splice variant was determined during EndoG over-expression in CaCo-2 cell line, after EndoG treatment of cell cytoplasm and nuclei and after nuclei incubation with EndoG digested cell RNA. hTERT alternative splicing was induced by 47-mer RNA oligonucleotide in naked nuclei and in cells after transfection. Identified long non-coding RNA, that is the precursor of 47-mer RNA oligonucleotide. Its size is 1754 nucleotides. Based on the results the following mechanism was proposed. hTERT pre-mRNA is transcribed from coding DNA strand while long non-coding RNA is transcribed from template strand of hTERT gene. EndoG digests long non-coding RNA and produces 47-mer RNA oligonucleotide complementary to hTERT pre-mRNA exon 8 and intron 8 junction place. Interaction of 47-mer RNA oligonucleotide and hTERT pre-mRNA causes alternative splicing.

GABA production and structure of gadB/gadC genes in Lactobacillus and Bifidobacterium strains from human microbiota.

Gamma-amino butyric acid (GABA) is an active biogenic substance synthesized in plants, fungi, vertebrate animals and bacteria. Lactic acid bacteria are considered the main producers of GABA among bacteria. GABA-producing lactobacilli are isolated from food products such as cheese, yogurt, sourdough, etc. and are the source of bioactive properties assigned to those foods. The ability of human-derived lactobacilli and bifidobacteria to synthesize GABA remains poorly characterized. In this paper, we screened our collection of 135 human-derived Lactobacillus and Bifidobacterium strains for their ability to produce GABA from its precursor monosodium glutamate. Fifty eight strains were able to produce GABA. The most efficient GABA-producers were Bifidobacterium strains (up to 6 g/L). Time profiles of cell growth and GABA production as well as the influence of pyridoxal phosphate on GABA production were studied for L. plantarum 90sk, L. brevis 15f, B. adolescentis 150 and B. angulatum GT102. DNA of these strains was sequenced; the gadB and gadC genes were identified. The presence of these genes was analyzed in 14 metagenomes of healthy individuals. The genes were found in the following genera of bacteria: Bacteroidetes (Bacteroides, Parabacteroides, Alistipes, Odoribacter, Prevotella), Proteobacterium (Esherichia), Firmicutes (Enterococcus), Actinobacteria (Bifidobacterium). These data indicate that gad genes as well as the ability to produce GABA are widely distributed among lactobacilli and bifidobacteria (mainly in L. plantarum, L. brevis, B. adolescentis, B. angulatum, B. dentium) and other gut-derived bacterial species. Perhaps, GABA is involved in the interaction of gut microbiota with the macroorganism and the ability to synthesize GABA may be an important feature in the selection of bacterial strains - psychobiotics.

[Apoptotic endonuclease EndoG induces alternative splicing of telomerase catalytic subunit hTERT and death of tumor cells]. / Éndonukleaza EndoG indutsiruet al'ternativnyi splaising kataliticheskoi sub"edinitsy telomerazy hTERT i gibel' opukholevykh kletok.

Telomerase activity is known to be regulated by alternative splicing of its catalytic subunit hTERT (human Telomerase Reverse Transcriptase) mRNA. Induction of non-active spliced hTERT leads to inhibition of telomerase activity. However, very little is known about the mechanism of hTERT mRNA alternative splicing. The aim of this study was to determine the role of apoptotic endonuclease EndoG in alternative splicing of hTERT and telomerase activity. Strong correlation was found between expression of EndoG and hTERT splice-variants in 12 colon cancer cell lines. Overexpression of EndoG in СаСо-2 cells downregulated the expression of active full-length hTERT variant and upregulated non-active spliced variant. Reduction of full-length hTERT caused downregulation of telomerase activity, dramatically shortening of telomeres length during cell divisions, converting cells to the replicative senescence state, activation of apoptosis and finally cell death. These data indicated the participation of EndoG in alternative splicing of mRNA of telomerase catalytic subunit, regulation of telomerase activity and cell fate.

[EFFECTS OF HYPOXIA AND GROWTH FACTORS ON THE ANGIOGENIC ACTIVITY OF MULTIPOTENT MESENCHYMAL STROMAL CELLS].

The effects of fetal calf serum (FCS) growth factor concentration and cell growth phase on production of angiogenic mediators by mesenchymal stromal cells (MSCs) at different O2 levels (20 and 5%) was studied. For this purpose vascular endothelial growth factor (VEGF-A) production was measured in MSC-conditioned medium (CM); besides, branching vessels as well as vessel end points (ramification) in the chorioallantoic membrane of Japanese quail eggs (Coturnix coturnix japonica) were counted following MSC-CM application. During the standard cultivation (20% O2; 10% FCS) the total number of vessels was 1.6 times higher comparing with hypoxic condition (5% O2; 10% FCS) due to increase in ramification, the number of branching vessels did not change. Maximal (double) increase in the total vessel number was observed when CM from MSCs after hypoxia plus serum deprivation was added. VEGF-A synthesis linearly increased with FCS concentration both at 20% and 5% O2. In all cases VEGF-A level was higher at hypoxia. No direct correlation between the VEGF-A concentration and total number of vessels was noted indicating that hypoxia possibly stimulates synthesis of additional angiogenic factors to enhance vascular growth despite the drastic serum deprivation. At 20% oxygen, exponentially growing MSCs showed the highest angiogenic activity and the ramification increased in 1.6 times. Depending on O2, MSCs produced angiogenic factors required at different stages of vascularization. Specifically, mediators of ramification were accumulated in the standard conditions (20% O2) and factors stimulating growth of branching vessels--in hypoxia.

[Antitumor enzyme L-lyzine-alpha-oxidase from Trichoderma harzianum Rifai F-180 and investigation of its action on L-lysine oxidation by capillar electrophoresis].

Trichoderma harzianum Rifai F-180, an organism producing the antitumor enzyme L-lysine-alpha-oxidase was cultivated and the enzyme was isolated and purified under the manufacturing conditions. The effect of L-lysine-alpha-oxidase on oxidation of L-lysine was investigated for the first time by capillary electrophoresis and the procedure conditions were developed. The reaction of L-lysine oxidative deamination is described and location of the reaction components picks on the elecrophoregrams was identified. The average rate of the catalytic reaction of L-lysine oxidation equal to 0.46 RU/min (7.7 x 10(-3) RU/sec) was determined. The use of the antitumor enzyme L-lysine-alpha-oxidase is recommended as a drug for the treatment of superficial tumors and tissue relative oxygen excess.

[Comparative evaluation of the effects of hypoxic gaseous mixtures varying in oxygen concentration on Japanese quail embryogenesis].

The paper presents the results of comparative characterization of the effects of low oxygen levels (10 ± 0.5 and 14.5 ± 0.5%) on developing organism. Four-day old embryos of the Japanese quail (Coturnix coturnix japonica) were chosen for the object of investigation as this is the age when avians acquire their organs and systems. Acute hypoxia (10 ± 0.5% oxygen) caused a general death of the embryos, and serious abnormalities of the eye and brain, and ectopy. Embryos that developed in the low-oxygen atmosphere (14.5 ± 0.5%) did not exhibit many of these morphological abnormalities and yet their growth was retarded apparently. Such abnormalities in acute hypoxia are ascribed to disturbance in development of extra-embryonic membranes, amnion in particular, desynchronization of morphogenetic processes and movement of embryo's tissue layers.

[Effect of ultrasonic treatment on the intensification of processes for obtaining amino acid mixtures from ethanol-assimilating yeasts]. / Vliianie ul'trazvukovoi obrabotki na intensifikatsiiu protsessa polucheniia aminokislotnykh smesei iz assimiliruiushchikh étanol drozhzhei.

We studied the effects of ultrasound treatment on an increase of the yield of amino acid mixtures from autolysates of Saccharomyces cerevisiae VKM-Y-2465 grown on ethanol. Adding 2% chloroform (activating agent), autolysis for 5-10 h, and ultrasound treatment of autolysates allowed us to obtain mixtures balanced in lysine, methionine, and tryptophan and containing up to 90% free amino acids.

[Effect of L-lysine-alpha-oxidase on skin carcinoma induced by methylcholanthrene in mice]. / Vozdeisvie L-lizin-al'fa-oksidazy na kartsinomu kozhi myshei, indutsirovannuiu metilkholantrenom.

Effect of enzyme L-lisyne-a-oxidase in complex with gel (Sakap) on carcinoma of the skin of mice induced by methylcholantrene was studied. Gel application once daily during 3 weeks decreased carcinoma dimensions on 40 per cent.

[Component activity of the isoenzyme spectra of Saccharomyces cerevisiae, Saccharomyces carlsbergensis and their hybrids]. / Aktivnost' komponentov izozimnykh spektrov fermentov u Saccharomyces cerevisiae, Saccharomyces carlsbergensis i poluchennykh gibridov.

The object of this work was to study the activity and the isozyme spectra of hexokinase (the triggering enzyme of glycolysis), glucose-6-phosphate dehydrogenase (the key enzyme of the pentose-phosphate shunt), malate dehydrogenase and isocitrate dehydrogenase (the enzymes of the citric acid cycle) and alcohol dehydrogenase (the enzyme involved in the first steps of ethanol oxidation) in Saccharomyces cerevisiae, race Ya, S. carlsbergensis, race 4228, and their hybrid 67. The parent organisms and their hybrid were shown to differ from one another in the qualitative composition and the activity of the isozyme spectra of the above enzymes.

[Comparative study of the spectra and activities of isoenzymes in haploid and autodiploid strains of Pichia pinus]. / Sravnitel'noe izuchenie spektrov i aktivnosti izozimov fermentov u gaploidnogo i avtodiploidnogo shtammov Pichia pinus.

The spectra and activities of isozymes of hexokinase and alcohol dehydrogenase (enzymes that catalyse glucose and ethanol assimilation), glucose-6-phosphate dehydrogenase (the key enzyme of the pentose phosphate shunt) and malate dehydrogenase (an enzyme of the tricarboxylic acid cycle) were comparatively studied in Pichia pinus haploid (MH4) and autodiploid (D4) strains. Differences in the qualitative composition of the isoenzyme spectra and activities suggest that the intensity and the role of the studied metabolic pathways and cycles differ between the haploid and autodiploid strains of Pichia pinus.