Influence of the Cultivation Conditions of the Glioblastoma Neurosphere on the Expression of MALAT1 and LINCROR Long Non-coding RNA Genes.

Glioblastoma multiforme (GBM) is the most aggressive malignant brain tumor. One of the reasons for the resistance of GBM to treatment is the extreme heterogeneity of the tumor and, in particular, the presence of cancer stem cells (CSCs) in the population of glioblastoma cells. In this work, we investigated the effect of conditions that reduce the proportion of CSCs in the GBM cell population on the levels of long noncoding RNAs (lincROR and MALAT1) involved in the formation of the phenotype of glioblastoma cancer stem cells. We have shown that culturing under conditions that cause a decrease in cell stemness (when fetal bovine serum is added to the culture medium) affected the content of these transcripts: in the cells of most of the analyzed lines, a decrease in the level of the positive stemness regulator lincROR and an increase in the content of MALAT1 were noted.

The Prognostic Significance of Spliceosomal Proteins for Patients with Glioblastoma.

Glioblastoma (GBM) is considered one of the most aggressive human cancers. Earlier, our group have demonstrated that alternative RNA splicing plays an important role in the regulation of the GBM phenotype. To continue this study, we analyzed the type of RNA splicing and the expression levels of the spliceosomal genes in a large number of tumor tissue samples and patient-derived GBM sphere lines. We demonstrated that the expression level of splicing factors allows dividing GBM patients into groups with different survival prognosis and also reflects the phenotype of the tumor. In addition, we identified the alternative splicing events that may regulate the GBM phenotype. Finally, we for the first time compared the expression profiles of the spliceosomal genes in different regions of the same tumor and identified splicing factors whose expression most significantly correlates with GBM patients' survival. Aforementioned data emphasize the important role of pre-mRNA splicing in GBM progression.

[Twin gestation with regressive partial hydatidiform mole and coexisting live fetus]. / Nerazvivayushchiisya chastichnyi puzyrnyi zanos v slitnoi dikhorial'noi platsente s normal'nym razvitiem vtorogo bliznetsa.

The case of dichorionic twin pregnancy is described, with a fused placenta, one part of which is represented by a tissue of partial hydatidiform mole (PHM) with signs of regression, the second part is a placenta of a common structure with a normal development of the second twin. The delivery took place at the term of 38 weeks with a live healthy girl weighing 3250 g. A single placental disc consisted of two fused placentas with a clear boundary between them. The placenta of a live-born girl was mature, with focal chorangiosis, the second part of the disc was represented by the PHM tissue with avascular giant bizarre villi, some of them with central cisterns, with stromal fibrosis, low proliferative activity of the villous trophoblast and a significant narrowing of the intervillous space. A genetic study was carried out on the material of paraffin blocks from two parts of the placental disc containing the tissue of the villous chorion, and the blood of the parents. Comparative analysis of DNA isolated from the paraffin block of PHM with the DNA of the parents revealed the presence of diandric dispermic triploidy. No chromosomal pathology was found in the placenta of a living girl. For hydatidiform mole in the case of multiple pregnancy, an increase in the volume of the affected placenta is characteristic compared to the normal placenta of the twin. In our observation, the presence in the placenta with PHM signs characteristic of placentas with antenatal fetal death, stromal fibrosis of the villi and low proliferative activity of the trophoblast suggests a regression of PHM.

THE INFLUENCE OF CONTRACEPTION ON VAGINAL MICROBIOCENOSIS CONDITION.

Aim - in complex to evaluate different types of female vaginal biocenosis on the background of contraceptives usage in order to detect implant-associated infection. Clinical status, features of vaginal microflora and formation of biofilms were assessed in 64 women of reproductive age, who had inert intrauterine device (IUD) of the second generation for a period of from 5 months to 8 years. pH of vaginal secretion was measured by the test strips CITOLAB (Farmasko, Ukraine). The condition of vaginal biocenosis was assessed according to «Femoflor-Screen¼, polymerase chain reaction in real time. The formation of biofilms was explored with the study of the ability of pure culture Gardnerellaе to adhesion on the surface of microplates and the addition of crystal violet indicator. Optical density was measured with the photometer «Multiskan EX-355¼ (Labsystems, China). These vaginal micribiota of the patients in control group were inconsistent with microbiocenosis of I and II type (normocenosis, intermediate type). II type of the vagina normocenosis was found in 29,4 % patients with long usage of contraception up to 12 month. III type of vaginal microbiocenosis was observed in 55, 8 % of examined patients within the duration of IUD use from 2 till 5 years. Microbiocenosis of women with IV type of smear was inconsistent with non-specific or hybrid vaginitis (IUD duration more than 5 years). Changes of total bacterial exudates, detection of opportunistic or pathogenic flora, especially for mixed bacterial associations for women with intra-uterine devices gives rise to vaginal biofilm formation. The type of vaginal biocenosis depends on the duration of IUC use. рН level of vaginal secretion, which conforms to the quantitative evaluation of microbiocenosis and the availability of lactobacteria in it can be used as an indicator for condition of vaginal biocenosis. pН level of vaginal secretion can be a screen control of vaginal biocenosis condition during intrauterine devices stay for the control of its durability in the uterus.

[CHARACTERISTICS OF LEVOFLOXACIN AND ITS CLINICAL APPLICATION (REVIEW)].

We analyzed of the effectiveness of levofloxacin in various diseases, the sensitivity of microorganisms to it and side effects. LF does not show carcinogenic, mutagenic and teratogenic activity. It is used effectively in the treatment of infections of the respiratory and genitourinary tract, skin and soft tissues, for the prevention of complications in surgical practice, as well as infections of many other localizations. Levofloxacin has a wide spectrum of action, including most gram-positive and gram-negative pathogens, including intracellularly located, unlike other fluoroquinolones, highly active against gram-positive cocci, including pneumococci which are resistant to penicillin, erythromycin. In addition, levofloxacin is more active against atypical pathogens. Doctors should clearly justife the appointment of LF in an adequate dose in each case, in order to avoid or reduce the occurrence of resistance of pathogens to it.

[Effect of arterial hypertension and hyperlipidemia on remodeling of articular cartilage and the development of osteoarthritis (experimental study).]

Osteoarthritis (OA) is a degenerative-inflammatory disease of the synovial joints associated with age, cardiovascular comorbidity, and other factors, based on cartilage (AC) and subchondral bone (SCB) damage. Recent studies have shown that age-related changes, cardiovascular diseases and OA may have a number of common molecular mechanisms. At the same time, the conditions and the degree of influence of arterial hypertension (AH) and hyperlipidemia (HL) on the tissues of the joints remain unclear. The purpose of the study is to study the effect of arterial hypertension and hyperlipidemia on the processes of cellular stress, remodeling of AC and the development of OA. An experimental study was carried out on 18 adult males of purebred guinea pigs (28-30 weeks old, weight 750-900 g). The 1st group (model AH) - 6 individuals, the 2nd (model with HL) - 6 individuals, the 3rd group (control) - 6 individuals. The results of the study allowed to establish that AH and HL have a direct effect on the tissues of the joints, causing cellular stress, manifested in changes in the morphofunctional characteristics of chondrocytes. Changes in the phenotype of cells leads to degradation of AC and SCB, ectopic angioproliferation. However, cardiometabolic factors influence AC remodeling processes in different ways. Thus, with isolated hypertension, hypertrophic differentiation of chondrocytes, destruction of articular cartilage, loss of cambial cells are observed. In HL, cell death processes, pathological mineralization of articular cartilage and enhanced pathological angiogenesis are observed. The greatest changes in articular cartilage are caused by the combination of AH and HL. With a combination of cardiometabolic factors, necrotic destruction of AC and replacement of SCB with osteopod-like matrix is observed.

[Placental mesenchymal dysplasia]. / Mezenkhimal'naia displaziia platsenty.

OBJECTIVE: To carry out a clinical and morphological analysis of 6 cases of placental mesenchymal dysplasia (PMD) that is not associated with Beckwith-Wiedemann syndrome. MATERIAL AND METHODS: Medical records, placental macroscopic and microscopic changes, histochemical (MSB staining) and immunohistochemical studies of placental tissue with antibodies against p57, CD34, smooth muscle actin, desmin, and Ki-67 were analyzed. RESULTS: Vascular anomalies in the chorionic plate and stem villi, the increased size and edema of the stem villi during normal formation of the terminal branches of the villous tree, the lack of proliferation of villous trophoblast were the typical signs of PMD and were noted in all cases. Comparison of the results of ultrasonography with the morphological pattern of the disease suggested that there were ultrasound signs that were typical of PMD. The characteristics of the course and outcomes of pregnancy in PMD were given. The features of morphological changes in the presence of PMD concurrent with preeclampsia were found. Significant variability in p57 expression in PMD was shown and variants of changes given. There were no substantial features of the expression of desmin and smooth muscle actin in PMD. CONCLUSION: MDP has typical morphological and ultrasound signs. The significant variability in the levels of chorionic gonadotropin and alpha-fetoprotein and in the expression of p57 does not allow their use in the differential diagnosis of PMD. The high incidence of thrombotic events in the intervillous space and fetal vessels, as well as intrauterine growth restriction, intrauterine hypoxia, and an impaired neonatal adaptation period in PMD should be taken into account when determining the management tactics for female patients and newborns.

Pseudogenes as Functionally Significant Elements of the Genome.

Pseudogene is a gene copy that has lost its original function. For a long time, pseudogenes have been considered as "junk DNA" that inevitably arises as a result of ongoing evolutionary process. However, experimental data obtained during recent years indicate this understanding of the nature of pseudogenes is not entirely correct, and many pseudogenes perform important genetic functions. In the review, we have addressed classification of pseudogenes, methods of their detection in the genome, and the problem of their evolutionary conservatism and prevalence among species belonging to different taxonomic groups in the light of modern data. The mechanisms of gene expression regulation by pseudogenes and the role of pseudogenes in pathogenesis of various human diseases are discussed.

[The prospects for the further development of the Belokurikha spa and health resort territory in the Altai Krai (region)].

We undertook a balneological survey of the Belokurikha spa and health resort territory with the purpose of distinguishing and identifying the potential health-improvement areas most promising for the extension and optimization of the therapeutic, tourist and recreational activities. The assessment was focused on the characteristic of the landscape and climatic conditions of the territory, the possibilities for the development of the existing resources of mineral waters and therapeutic muds as well as for the discovery of the potential new ones. The recommendations are proposed to promote the development of different forms of tourism with special reference to its medical and health-improvement aspects. It is suggested that the territory of the «Belokurikha¼ resort¼, «Belokurikha-2¼ and «Belokurikha-3¼ health-improvement areas should be integrated into a single spa-and-health resort district of federal importance.

Functions of noncoding sequences in mammalian genomes.

Most of the mammalian genome consists of nucleotide sequences not coding for proteins. Exons of genes make up only 3% of the human genome, while the significance of most other sequences remains unknown. Recent genome studies with high-throughput methods demonstrate that the so-called noncoding part of the genome may perform important functions. This hypothesis is supported by three groups of experimental data: 1) approximately 10% of the sequences, most of which are located in noncoding parts of the genome, is evolutionarily conserved and thus can be of functional importance; 2) up to 99% of the mammalian genome is being transcribed forming short and long noncoding RNAs in addition to common mRNA; and 3) mutations in noncoding parts of the genome can be accompanied by progression of pathological states of the organism. In the light of these data, in the review we consider the functional role of numerous known sequences of noncoding parts of the genome including introns, DNA methylation regions, enhancers and locus control regions, insulators, S/MAR sequences, pseudogenes, and genes of noncoding RNAs, as well as transposons and simple repeats of centromeric and telomeric regions of chromosomes. The assumption is made that the intergenic noncoding sequences without definite/clear functions can be involved in spatial organization of genetic loci in interphase nuclei.

[Possibilities and effects of telomerase activation].

In this review we briefly describe recent knowledge of telomerase (predominately human telomerase) activity regulation mechanisms. We also point telomerase complex components localization in cells and discuss the enzyme activities that are independent of telomere elongation. The paper includes the overview of human diseases correlating with reduced telomerase activity, short telomeres and rapid telomeres shortening. We describe in details the possibilities of exogenous hTERTgene transcription activation by different natural and synthetic compounds as well as hTERTgene transfection effects. Such exogenous activation cause increasing proliferative potential of the cells and might be used in cell therapy. It must be noticed that elevated hTERT gene expression, especially in the case of hTERTgene transfection, might be the cause of cell malignesation. In this regard strict constraining criteria in medical application of different methods of telomerase activation must be developed.

[Pseudogene PTENP1 5'-region methylation in endometrial cancer and hyperplasias].

Genetic mutations in tumor suppressor gene PTEN are often detected in malignant human cells and these genomic changes are especially characteristic ofendometrial cancer. In our previous researches we assumed that alternative epigenetic mechanism of PTEN inactivation trough promoter methylation may exist in endometrial cancer. Moreover, PTENP1 pseudogene has recently been shown to play a role in positive regulation of PTENgene expression. Taking into account these facts, we analyzed PTEN and PTENP1 methylation status in endometrial hyperplasia and cancer. It was demonstrated that PTENgene promoter was not methylated but PTENP1 was methylated in 11 of 18 endometrial cancers and in 5 of9 endometrial hyperplasias. We can assume that PTENP1 methylation may inhibit transcription of this gene and also PTEN gene transcription trough RNA interference in accordance with ceRNA theory. Thus, aberrant suppression of PTENP1 transcription can play a role in endometrial cancer pathogenesis.

Temperature dependence of growth-sector-dependent Raman spectra of boron-doped diamonds synthesized at high-pressure high-temperature.

Single crystals of boron-doped diamond (BDD) were synthesized by the temperature gradient method in high-pressure and high-temperature conditions in the Fe-Al-B-C system, and multisectoral diamond plates were extracted. Temperature-dependent (77-600 K) high-resolution Raman spectroscopic studies have been carried out to investigate the behavior of anharmonic phonon decay in the {001}, {113}, and {111} growth sectors of multisectoral diamond plates with different content of boron impurities (⩽80 ppm) and compare with the data for undoped IIa diamond. Micro-Fourier transform infrared spectroscopy was used to estimate the spatial distribution of uncompensated boron impurity[Na-Nd]in BDD plates by analyzing boron-related absorption peaks. The plates were shown to have non-uniform growth-sector-dependent content of uncompensated boron impurity in the range from 1.1 × 1018to 1.4 × 1019cm-3. The effects of anharmonic decay (damping) of optical phonons in BDD are studied by modeling the temperature dependence of phonon frequency and linewidth of the diamond's F2gand boron-induced vibrational modes. The extrapolated zero-temperature optical phonon linewidth and frequency and the anharmonic nature of their linear relationship are determined as a function of the growth sector and boron doping. The predominant mechanisms and parameters of the anharmonic decay of optical phonons are determined, which is of fundamental importance for the thermal conductivity of semiconductor materials. The anharmonic phonon decay remained the predominant process at higher temperatures, irrespective of the doping level.

[Polymorphism of 5,10-methylenetetrahydropholate reductase, prothrombin, and coagulation factor V genes in young patients with ischemic stroke].

The study included 142 patients (87 women, 55 men) (mean age 36.2 +/- 8.3 yr) after ischemic stroke caused by dissection of cerebral arteries (D) (n = 37), anti-phospholipid syndrome (APS) (n = 55) or cardiogenic embolism (CE) (n = 11). Stroke of unknown origin (cryptogenic) was diagnosed in 39 patients. Mutations of 5,10-methylenetetrahydropholate reductase (MTGPR), prothrombin, and coagulation factor V genes were documented by PCR in 38, 0, 3% of D cases, 55.9, 9, 13% of APS cases, 73, 9, 0 CE cases, 57, 5, 0% of cases with cryptogenic stroke compared with 43, 0, 0% in controls. Mutations in MTGPR gene in CE cases, prothrombin gene in APS and CE cases, coagulation factor V gene in APS cases occurred more frequently than in control (p < 0.05). They, were more frequent in APS/CE than in D (p < 0.05). Mutation rate in cryptogenic stroke was not significantly different from that in control (p < 0.05). It is concluded that the above mutations are not involved in pathogenesis of cryptogenic stroke, whereas those of prothrombin and coagulation factor V genes may enhance the thrombogenic potential in APS and CE patients. The role of MTGPR gene mutation in pathogenesis of cardiogenic stroke needs clarification.

[A new view on the problem of nonneoplastic stenosis of the major duodenal papilla. Results of a prospective investigation].

The article contains results of a continuous prospective investigation of the normal course of the major duodenal papilla stenosis in 167 patients. It was revealed that the course of the disease was benign, pain syndrome became inconsiderable in the course of time, the degree of changes in biochemical analysis of blood was not significant. The development of complications (choledocholithiasis, acute pancreatitis, jaundice) was noted in 6-14% of the patients, depending on the presence or absence of GID. The data obtained allow suggestion of an algorithm of managing the patients.

[Comparative characteristics of antioxidant status in women with diabetes type 2 of different age groups].

The purpose of this study was to evaluate the metabolic processes in women with diabetes mellitus type 2 of different age groups. It is established that hyperglycemia in aged women is characterized by the development of pronounced oxidative stress, which is the result of changes in the primary structure of protein molecules due to non enzymatic glycosylation of amino acid residues in the active sites. It is known that observed depletion of reduced glutathione pool is associated with high risk of genotoxicity, because it correlates with activation of mitochondrial, chromatin dysfunction and fragmentation of the DNA. In addition, hydroperoxides of polyunsaturated fatty acids formation leads to necrosis and apoptosis. It can be assumed that the diabetes mellitus type 2 triggers processes of apoptosis, which leads to the activation of aging programs and increase the mortality of patients. Obviously, the change in the concentration of thiol antioxidants, as well as the change in concentration of LPO molecular products may be one of the criteria for evaluation of aging and the efficiency of the treatment of patients.

[Pregnancy in women with heart defects: outlook on the problem].

Modern approach to management of pregnancy and delivery in women with heart defects is presented in this paper. The 3-level system of observation of this contingent of patients is based on stratification of risk of development of cardiological complications which allows to form an algorithm of physicians actions, and to optimize tactics of ambulatory and hospital stages of treatment. Stratification of risk is supplemented with quantitative echocardiographic parameters which reflect hemodynamic overload of the myocardium and facilitate work of a practical physician. Special consideration is given to pregnant women subjected to palliative cardiac surgery. If contractile function is preserved these women have good prognosis relative to prolongation of pregnancy and unassisted delivery.

[Gastric carcinoid--modern approaches to diagnosis and treatment].

Gastric carcinoid tumors are rare, but are increasing in incidence. This article reviews what is known about the classification, epidemiology, tumor pathogenesis and management of patients. Additionally we present the results of our own 10-years observation after endoscopic and surgery treatment of this tumors.

[Peculiarities of metabolic rearrangement of gas-transporting function of the blood in the pregnant women under the influence of medical ozone applied in obstetrical practice].

This clinical and laboratory study was designed to evaluate the influence of an ozone-oxygen mixture on the characteristics of blood oxygen supply in pregnant women presenting with chronic placental insufficiency. The study has demonstrate that dosed ozone application modulates the oxygen-transporting function of the blood and promotes adaptive capacities of the organism by activation of aerobic metabolism and stabilization of cellular membranes. It is concluded that the proposed method can be recommended for the treatment of women with the complicated course of pregnancy in order to improve their endurance of and resistibility to the action of hypoxemia.

[Oxygen-dependent processes in the blood cells from children with arterial hypertension concomitant with biliary dyskinesia].

Age individuality is characterized by an imbalance of the molecular mechanisms of antioxidant defense in adolescents with arterial hypertension and biliary dyskinesia, as documented by an enzyme imbalance of the first line of antioxidant defense and H2O, accumulation, by a substantial increase in glutathione peroxidase activity, and by inhibition of the activity of glutathione-dependent enzymes. The considerable rise of 2,3-diphosphoglycerate suggests tissue hypoxia. With this, enhanced neutrophil elastase activity causes damage to the structural components of vascular wall connective tissue, resulting in the development of endothelial dysfunction.