Strategies to enhance stress tolerance in lactic acid bacteria across diverse stress conditions.

Lactic acid bacteria (LAB) hold significant importance in diverse fields, including food technology, industrial biotechnology, and medicine. As basic components of starter cultures, probiotics, immunomodulators, and live vaccines, LAB cells resist a variety of stressors, including temperature fluctuations, osmotic and pH shocks, exposure to oxidants and ultraviolet radiation, substrate deprivation, mechanical damage, and more. To stay alive in these adversities, LAB employ a wide range of stress response strategies supported by various mechanisms, for example rearrangement of metabolism, expression of specialized biomolecules (e.g., chaperones and antioxidants), exopolysaccharide synthesis, and complex repair and regulatory systems. LAB can coordinate responses to various stressors using global regulators. In this review, we summarize current knowledge about stress response strategies used by LAB and consider mechanisms of response to specific stressful factors, supported by illustrative examples. In addition, we discuss technical approaches to increase the stress resistance of LAB, including pre-adaptation, genetic modification of strains, and adjustment of cultivation conditions. A critical analysis of the recent findings in this field augments comprehension of stress tolerance mechanisms in LAB, paving the way for prospective research directions with implications in fundamental and practical areas.

[Hypermethylation of miR-107, miR-130b, miR-203a, miR-1258 Genes Associated with Ovarian Cancer Development and Metastasis].

It is known that microRNAs (miRNAs) are able to dynamically regulate gene expression. At the same time, methylation can reduce expression of miRNA encoding genes and, therefore, reduce their inhibitory effects on mRNAs of target genes, including those of oncogenes, that promoting the development of tumors of different localization. The role of miRNA hypermethylation in the pathogenesis of ovarian cancer is not completely understood; so we conducted a search for new hypermethylated and potentially suppressor miRNA genes in ovarian tumors. Four new miRNA genes (MIR-107, MIR-130b, MIR-203a, MIR-1258) commonly hypermethylated (28-52%) in tumor tissues vs 4-7% in paired histologically normal tissues, p < 0.01, were identified in a representative set of 54 ovarian cancer samples using methylation-specific PCR. It was shown that hypermethylation of MIR-130b, MIR-203a, and MIR-1258 genes is significantly (p < 0.05) associated with metastasis of ovarian cancer. These results suggest the involvement of four miRNAs (miR-107, miR-130b, miR-203a, and miR-1258) and hypermethylation of their encoding genes in the pathogenesis of ovarian cancer.

[CHARACTERISTICS OF FUNGAL STRAINS PRODUCING THERMOSTABLE XYLOGLUCANASES FROM THE RUSSIAN NATIONAL COLLECTION OF INDUSTRIAL MICROORGANISMS.]

Fungal strains degrading plant biomass available from the Russian National Collection of Industrial Microorganisms (VKPM) have been screened for the xyloglucanase activity. Under conditions of submerged cultivation, the thermophilic strains Sporotrichum thermophile VKPM F-972, Myceliophthora thermophila VKPM F-244, and Sporotrichum pruinosum VKPM F-235 produced extracellular xyloglucanases with optimal activity at 60°C, pH 5.0. 88-100% of the initial enzyme activity was retained after l-h incubation at 50°C; 79-84% of the activity was retained after l-h incubation at 60°C. S. thermophile VKPM F-972, M. thermophila VKPM F-244, and S. pruinosum VKPM F-235 strains may be used as the gene sources for construction of highly active producers of the thermostable xyloglucanases.

Upregulation of RHOA and NKIRAS1 genes in lung tumors is associated with loss of their methylation as well as with methylation of regulatory miRNA genes.

Methylation of CpG-islands in promoter regions as well as interaction of miRNAs with messenger RNAs of target genes are related to multilayer mechanisms regulating gene expression. The goal of this study was to assess a possibility for miRNA gene methylation to influence indirectly activation of their target genes in lung tumors. By using a unified collection of samples of non-small cell lung cancer, it was demonstrated that elevated levels of mRNA for RHOA and NKIRAS1 genes were significantly (Spearman rank correlation, P < 10(-11)) associated both with loss of methylation in their CpG-islands and methylation in a number of miRNA genes, which, according to the miRWalk database, were predicted to possess regulatory functions. Novel potential regulatory miRNAs for RHOA (miR-9-1/-3, -34b/c, -129-2, -125b-1, -375, -1258) and NKIRAS1 (miR-34b/c, -129-2, -125b-1, -193a, -124a-1/-2/-3, -212, -132) genes in lung cancer were identified.

Methylation of miRNA genes and oncogenesis.

Interaction between microRNA (miRNA) and messenger RNA of target genes at the posttranscriptional level provides fine-tuned dynamic regulation of cell signaling pathways. Each miRNA can be involved in regulating hundreds of protein-coding genes, and, conversely, a number of different miRNAs usually target a structural gene. Epigenetic gene inactivation associated with methylation of promoter CpG-islands is common to both protein-coding genes and miRNA genes. Here, data on functions of miRNAs in development of tumor-cell phenotype are reviewed. Genomic organization of promoter CpG-islands of the miRNA genes located in inter- and intragenic areas is discussed. The literature and our own results on frequency of CpG-island methylation in miRNA genes from tumors are summarized, and data regarding a link between such modification and changed activity of miRNA genes and, consequently, protein-coding target genes are presented. Moreover, the impact of miRNA gene methylation on key oncogenetic processes as well as affected signaling pathways is discussed.

[The effectiveness of comprehensive rehabilitation of the patients presenting with coronary heart disease with the application of underwater shower massage and electrical stimulation].

The underwater shower massage and electrical stimulation were applied to treat 120 patients presenting with coronary heart disease. The estimation of the effectiveness of this treatment with the use of functional diagnostic methods revealed its psychocorrective effect manifested in the form of reduced frequency and duration of painful and painless angina seizures, restricted sympathetic influences on the heart as well as normalized sympathovagal balance, lipid status, antioxidative system, central and peripheral hemodynamics, improved systolic and diastolic function of the myocardium, enhanced tolerability of mental, emotional, and physical workload. The study allowed the predictors of the effectiveness of therapeutic and health-promoting measures to be identified.

[Methylation profile of group of miRNA genes in clear cell renal cell carcinoma; involvement in cancer progression].

MicroRNA regulates gene expression, is involved in many cellular processes, and plays an important role in the development of cancer. The regulation of the expression of miRNA genes can be achieved by methylating their CpG islands, which is shown in different types of tumors. The methylation of miRNA genes in clear cell renal cell carcinoma (CCRCC) has mainly been studied for the miR-9 and miR-34 families. The methylation of six miRNA genes (miR-124a-2, -124a-3, -9-1, -9-3, -34b/c, -129-2) was investigated with the use of representative set of CCRCC samples (46 cases). Methylation of three genes miR-124a-2, -124a-3, and -129-2 was studied in kidney tumors for the first time. Methylation analysis was performed using methyl specific PCR. It is shown that the frequency of methylation of six genes (miR-124a-2, -124a-3, -9-1, -9-3, -34b/c and -129-2) was significantly higher in tumor samples than in samples of histologically normal tissue (P < 3 x 10(-5) by Fisher's exact test). These results suggest the properties of tumor suppressors for the six miRNA genes indicated in CCRCC. We also found correlations between the methylation frequency of some miRNA genes and signs of the progression of CCRCC (tumor size, clinical stage, loss of differentiation, and metastasis).

[Modern combined technologies for the rehabilitation of the patients presenting with arterial hypertension and its prevention].

The present review describes the modern methods for the combined treatment and rehabilitation of the patients presenting with arterial hypertension and their influence on the key pathogenetic components of this condition in accordance with the currently accepted theories of its origin and evolution. The discovery of specific molecular and cellular mechanisms of the therapeutic action of certain factors that reflect peculiar and unique character of their inherent physical energy provided a basis for the development of highly effective methods for the treatment and prevention of arterial hypertension and rehabilitation of the patients suffering this disease. The most comprehensive clinical effect is produced by the application of scientifically sound therapeutic systems; exclusion of certain therapeutic physical factors from these modalities results in s significant reduction of their curative effectiveness.

[Novel miRNA genes methylated in lung tumors].

MicroRNAs play an important role in the regulation of expression of many genes and are involved in carcinogenesis. The regulation of miRNA gene expression can involve the methylation of promoter CpG islands. In this work, the methylation of six miRNA genes (mir-107, mir-125b-1, mir-130b, mir-137, mir-375, and mir-1258) in non-small-cell lung cancer (NSCLC) was studied for the first time by methylation-specific PCR using a representative set of specimens (39 cases). Four new genes (mir-125b-1, mir-137, mir-375, and mir-1258) methylated in primary NSCLC tumors were identified with frequencies of 56, 31, 56, and 36%, respectively. The frequencies of miRNA promoter methylation in DNA of tumors and histologically normal tissues differed significantly (P < or = 0.05 by Fisher's test). In lung tissues of 20 donors without a history of cancer, these genes were only methylated in a few cases. It was also shown that the previously unstudied promoter CpG islands of mir-107 and mir-130b were not methylated in NSCLC. The frequencies of mir-125b-1 and mir-137 methylation were shown for the first time to correlate with NSCLC progression (clinical stage and metastasis).

[Methylation of some miRNA genes is involved in the regulation of their target genes RAR-beta2 and NKIRAS1 expression in lung cancer].

To date, there are more than two thousand human miRNAs, each of them may be involved in the regulation of hundreds of protein coding target genes. Methylation of CpG-islands, in turn, affects miRNAs gene expression. Our aim was to evaluate the role of methylation in the regulation of miRNA gene expression and, consequently, in the regulation of expression of target genes in primary lung tumors. Using a common collection of non-small cell lung cancer samples we performed a comprehensive study, including analysis of the methylation status and expression levels of some miRNA genes and their potential target genes on chromosome 3: RAR-beta2 and NKIRAS1. Increased frequency of methylation in lung tumors compared to histologically normal tissue was revealed for miR-9-1 and miR-34b/c genes with significant statistics (P < or = 0.05 by Fisher exact test) and for miR-9-3 and miR-193a was marginally significant (P < or = 0.1). Significant correlation was revealed between alterations of methylation and expression level of miR-9-1 gene (P = 5 x 10(-12) by Spearman) in the lung tumors, this suggests the role of methylation in the regulation of expression of this miRNA genes. Besides, a statistically significant negative correlation (P = 3 x 10(-12)-5 x 10(-13) by Spearman) was found between alterations of expression levels of miR-9-1 and miR-17and RAR-beta2 target gene and also between expression level alterations of miR-17 and NKIRAS1 that was not previously analyzed. The inverse relationship between expression levels of miRNA genes and their target genes is consistent with the known mechanism of suppression of protein coding genes expression under the action of miRNAs. For the first time significant correlations (P = 3 x 10(-10)-4 x 10(-13) by Spearman) were shown between alterations of methylation status of miRNA genes (miR-9-1, miR-9-3, miR-34b/c, miR-193a) and the expression level of RAR-beta2 target gene and between alterations of methylation status of miR-34b/c, and miR-193a and the expression level of NKIRAS1 target gene in the primary lung tumors, which suggests the possibility of indirect effects of methylation of miRNA genes on expression level of target genes.

[Potentials of combined non-medication therapy of arterial hypertension associated with ischemic heart disease].

The objective of the present work was to develop methods and a scientifically sound rationale for the optimization of clinical results of the treatment of patients presenting with arterial hypertension (AH) associated with coronary heart disease (CHD) using red and infra-red (IR) low-intensity laser irradiation. The study included a total of 90 patients of working age (62.2% of men and 37.8% of women) having AH associated with coronary heart disease and abnormal blood lipid spectrum. They were allocated to three groups each comprising 30 patients matched for age, sex, severity of clinical conditions, and character of maintenance medicamental therapy. Patients of group 1 were treated by intravenous blood irradiation with laser light; those in group 2 were given similar treatment in combination with external low-intensity IR laser therapy; patients of group 3 received basal medicamental therapy. The study allowed the clinical value of different variants of low-intensity laser therapy to be assessed as regards efficiency of correction of hypersympaticotony, disturbances of lipid metabolism, and refractoriness to pharmaceutical products as well as improved tolerance of physical exercises, anti-hypertensive, anti-ischemic, and anti-anginal effects, diastolic function, and quality of life as targets of the comprehensive rehabilitative program designed for patients with AH associated with coronary heart disease. An optimized therapeutic modality is proposed for patients presenting with these pathologies based on the application of the low-intensity laser irradiation regime characterized by mutual amplification and complementary action of red and infrared lights.

[Analysis of free-radical products of near-UV irradiation at 77 K of frozen adenine and adenosine diphosphate solutions containing inorganic phosphate].

Irradiation by near-UV light at 77 K of aqueous solutions of inorganic phosphate in weakly acidic conditions in the presence of the photosensitizers adenine and adenosine diphosphate results in the formation of free radicals of these compounds, photosensitized free radicals of phosphate itself, and H* and OH* radicals. The relative concentrations of free radical products were estimated by the analysis of total ESR signals registered in the region of g = 2.00 in the photosystems Ade + Pi and Adphi + Pi using the original computer program of ESR spectra simulation.

[Free radicals photosensitized by near-UV radiation in aqueous adenine solutions in the presence of inorganic phosphate at 77 K].

It was shown that irradiation of aqueous adenine solutions in the presence of inorganic phosphate by near UV at 77 K results in the production of OH* free radicals catalyzed by phosphate and photosensitized formation of phosphate radicals and atoms H*.

Estimation of liposome integrity by 1H-NMR spectroscopy.

A fast and simple method of 1H-NMR spectroscopic control of liposome membrane integrity is suggested. The method is based on the redistribution of intensities between two singlet 1H-NMR signals--from intraliposomal marker compound (nitrilotriacetic acid sodium salt, 1H-NMR signal at 4 p.p.m.) and from its complex with Eu3+ added to the external medium (NMR signal of the complex at - 1 p.p.m.). The method permits registration of the kinetics of liposome destruction under the action of detergent or serum. It is shown that the presence of cholesterol in the membrane makes it more stable.