Structure and Functions of the Mediator Complex.

Mediator is a key factor in the regulation of expression of RNA polymerase II-transcribed genes. Recent studies have shown that Mediator acts as a coordinator of transcription activation and participates in maintaining chromatin architecture in the cell nucleus. In this review, we present current concepts on the structure and functions of Mediator.

Randomised phase III trial of vinflunine plus capecitabine versus capecitabine alone in patients with advanced breast cancer previously treated with an anthracycline and resistant to taxane.

Background: Capecitabine is an approved standard therapy for anthracycline- and taxane-pretreated locally advanced or metastatic breast cancer (BC). Vinflunine has demonstrated single-agent activity in phase II studies in this setting and activity and tolerability when combined with capecitabine. We compared the combination of vinflunine plus capecitabine (VC) with single-agent capecitabine. Patients and methods: Patients with locally recurrent/metastatic BC previously treated or resistant to an anthracycline and resistant to taxane therapy were randomly assigned to either vinflunine (280 mg/m2, day 1) plus oral capecitabine [825 mg/m2 twice daily (b.i.d.), days 1-14] every 3 weeks (q3w) or single-agent oral capecitabine (1250 mg/m2 b.i.d., days 1-14) q3w. The primary end point was progression-free survival (PFS) assessed by an independent review committee. The study had 90% power to detect a 30% improvement in PFS. Results: Overall, 770 patients were randomised. PFS was significantly longer with VC than with capecitabine alone [hazard ratio, 0.84, 95% confidence interval (CI), 0.71-0.99; log-rank P = 0.043; median 5.6 versus 4.3 months, respectively]. Median overall survival was 13.9 versus 11.7 months with VC versus capecitabine alone, respectively (hazard ratio, 0.98; 95% CI, 0.83-1.15; log-rank P = 0.77). No difference in quality of life was observed between the two treatment arms. The most common adverse events (NCI CTCAE version 3.0) in the combination arm were haematological and gastrointestinal. Grade 4 neutropenia was more frequent with VC (12% versus 1% with capecitabine alone); febrile neutropenia occurred in 2% versus 0.5%, respectively. Hand-foot syndrome was less frequent with VC (grade 3: 4% versus 19% for capecitabine alone). Peripheral neuropathy was uncommon in both arms (grade 3: 1% versus 0.3%). Conclusions: Vinflunine combined with capecitabine demonstrated a modest improvement in PFS and an acceptable safety profile compared with capecitabine alone in patients with anthracycline- and taxane-pretreated locally recurrent/metastatic BC. ClinicalTrials.gov: NCT01095003.

[The laryngeal condition in the patients presenting with euthyroid multinodular goiter as evaluated by direct laryngoscopy]. / Sostoianie gortani u bol'nykh éutireoidnym mnogouzlovym zobom po dannym priamoi laringoskopii.

The objective of the present study was to improve diagnostics of disturbances in the laryngeal condition of the patients presenting with euthyroid multinodular goiter with special reference to the condition of the upper respiratory passages. The preoperative characteristic of 398 patients included the description of the predominant clinical syndrome (the neoplastic and compressive variants) and specification of the morphological type of the disease to promote the decrease of operative activity with respect to colloidal proliferative goiter. Direct laryngoscopy was employed in addition to the traditional diagnostic techniques during both the surgical operation and the follow-up observation. Hoarseness developed in the postoperative period in 13.4% to 16.9% of the patients. Postoperative transient and persistent laryngeal paresis was documented in 1.5% and from 1.5% to 4.2% of the cases respectively. Direct laryngoscopy revealed the symptoms of laryngitis and laryngotracheitis as well as foci of leukoplakia, laryngeal cysts and nodules, besides disturbances in the mobility of the vocal chordae. It is concluded that the above pathological changes in the laryngeal structures should be identified during the preoperative examination of the patients presenting with euthyroid multinodular goiter while direct laryngoscopy must be included in the program of postoperative supervision.

Super-Resolution Microscopy in Studying the Structure and Function of the Cell Nucleus.

In recent decades, novel microscopic methods commonly referred to as super- resolution microscopy have been developed. These methods enable the visualization of a cell with a resolution of up to 10 nm. The application of these methods is of great interest in studying the structure and function of the cell nucleus. The review describes the main achievements in this field.

[The results of monitoring of antigen types of rotaviruses of group A on the territory of the Russian Federation in 2011-2015.]

The data of seasonal monitoring are presented concerning antigen types of rotaviruses group A circulating on the territory of the Russian Federation in the periods of seasonal uprising of morbidity in 2011-2015. Annually, the study included from 10 to 12 subjects of the Russian Federation with annual testing from 444 to 728 samples from children aged younger than 5 years with acute infection diarrhea. In the seasons of 2011-2012, 2012-2013, 2013-2014, 2014-2015 the most prevalent [P] G types of rotaviruses correspondingly made up to: G4[P]8 -50.2%-36.5%-43.8%-1.6%; G1[P]8 - 26.6%-14.3%-27.3%-22.5%; G3[P]8 - 4.4%-23.7%-4.2%-2.0%; G9[P]8 - 4.3%-5.3%-10.1%-7.1%; G2[P]4 - 7.7%-7.9%-9.0%-3.1%. The expressed territorial irregularity of prevalence of antigen types of retroviruses was observed.

[RESULTS OF USAGE OF ETHANOL DESTRUCTION AND MAGNETIC IRRADIATION IN PATIENTS WITH BENIGN NODULAR THYROID TUMORS].

The article presents an experience of treatment of 67 patients with benign thyroid tumors with application of ethanol destruction using guidance of ultrasonic imaging. It was stated, that complementary application of therapeutic laser and magnetic actions allowed shortening the terms of reduction of nodular tumors and smoothing out the ethanol effects, decreasing the possibility of specific complications, shortening the terms of general treatment of the patients and minimizing general costs.

[Monitoring of influenza and other respiratory diseases causative agents in children, hospitalized with community-acquired pneumonia in 2012-2013 epidemic season].

AIM: Study the circulation of respiratory viruses in children with community-acquired pneumonia (CAP) during the period from October 2012 to May 2013. MATERIALS AND METHODS: 136 children with CAP aged from 3 months to 16 years with ARI symptoms at the disease debut were studied. RNA/DNA of influenza A, B, parainfluenza (PI); adeno-, rhino-, RS-viruses, corona-, metapneumo- (MPV) and bocaviruses were detected in nasopharynx smears by PCR with hybridization-fluorescent detection in real time. Antibodies against influenza viruses A/H1N1/pdm09 California/07/09, epidemic reference strains of influenza viruses A/H1N1/Brisbane/59/07, A/ H3N2/Victoria/361/201 1, B/Wisconsin/1/10, against PI viruses type 1, 2, 3 were determined in paired sera by HAI. RESULTS: In February-March 2013 the number of children protected by antibodies against influenza decreased, and circulation of influenza viruses A/H3N2 and A/H1N1/ pdm09 was detected. Rhinoviruses and PI viruses were determined throughout the epidemic season, bocavirus and adenoviruses--during the autumn-winter period, RS-virus and MPV--during winter-spring. Coronaviruses were not detected. The peak of virus detection was established in February when the threshold of influenza and ARI morbidity was exceeded. The main pathogens of children of the first 3 years of life are rhinoviruses, RS-virus, PI viruses and bocavirus. RS-virus infection at the debut of CAP in children younger than 3 years in 55.5% of cases is associated with the development of broncho-obstructive syndrome. Bocavirus infection in 50% of cases progresses with laryngo-tracheitis and bronchiolitis. CONCLUSION: The fraction of viruses in etiologic structure ofARI in children varies depending on immune layer, season and age of children. Etiology of viral infection at the debut of CAP could only be proven using specialized laboratory studies.

[Medical-epidemiological and methodological approaches to forecasting indicators of adolescent health in the modern period].

Data on features of formation of health of different groups of the population in the development process of the country and its regions are an important information base for the development and implementation of measures for the maintenance and strengthening ofpublic health. The aim of the study was to determine and predict trends in indicators of primary disease incidence and prevalence among adolescent population over the next 10 years. Materials provided to analyze the data of the consolidated annual report forms .o 12 "The number of registered diseases in patients living in the service area of the medical establishment"for showing trends over the 10-year period (2001-2010). Calculation and analysis of dynamics were integral coefficients, determined the level of adolescent health violations from "low" to "high". Incidence rate of incidence and prevalence of disease were evaluated by calculating the dynamic coefficient of determination R approximation. The article presents results of forecasting over the next 10 years in the adolescent population (15-17years) changes the primary indicators incidence and prevalence of diseases. This technique was used by the authors. In view of the projected increase in detected trends data set of indicators at 40, 7-50,2%. Sex predicted dynamics of increase in juniors at 42,1-58.9%, the girls - the 31,2-49,6%. These data characterize negatively predicted for the near future a regional trend in incidence rates in the analyzed populations. They must be taken into account when deciding on the preservation and strengthening of health of the rising generation, including - in shaping regional social programs and health care programs.

[Regional ecological and social and economic aspects of morbidity of the teenage population residing in industrial centres].

It is presented the characteristic changes of atmospheric air pollution and morbidity of adolescents aged 15-17years living in urban environments with different specificity of industrial enterprises for the 10-year period. We investigated the relationship dynamics of the indicators of the health of modern teenagers with socio-economic indicators of regional development. A predictive model for the regression rate of absolutely healthy adolescents (group health I) is worked out and set the share of influence on this indicator factors included in the regression model.

[Molecular components of JAK/STAT signaling pathway and its connection with transcription machinery].

JAK/STAT is one of the conservative signaling pathways in higher eukaryotes which plays a critical role in different ontogenesis processes. This article gives a review of the pathway structure at the molecular level, mainly in a model organism Drosophila. There are data about relationship of the signaling pathway and transcription machinery of higher eukaryotes.

[Activation of JAK/STAT signaling pathway in S2 Drosophila melanogaster cell culture].

JAK/STAT signaling pathway plays a critical role in different ontogenesis processes of higher eukaryotes. Fruit fly drosophila is a handy model system used to study this pathway since major components of the pathway are represented by unique factors. This article describes the usage of Drosophila melanogaster S2 cells in studies of the pathway's target genes activation. We showed that S2 cells contain plenty of STAT protein which migrates into nucleus under cells treatment with pervanadate. Then we demonstrated that under pervanadate action STAT protein along with other transcription factors is recruited onto regulatory sequences of target genes and activates their transcription.

[Biological functions of Jak/Stat signaling pathway in Drosophila].

The basic biological processes under the control of the Jak/Stat signaling pathway in Drosophila are reviewed. As shown, the fruit fly Drosophila melanogaster is a very convenient model organism for investigation of Jak/Stat functions in various aspects of ontogenesis.

[Drosophila melanogaster imaginal discs mitotic anomalies induced by tumor-supressor dlg silencing construction].

Mitosis, cytokinesis and nuclear texture of wing imaginal discs cells silenced by UAS-RNAi-dlg construct induced by 1096-Ga 14 driver were studied. The silencing construct contains coding region of dlg gene and the complementary region. Further, this RNA hairpin (Dietzl et al., 2007) is processed by endogenous protein Dicer and the resulting RNA fragments silence mRNA dlg. Tumor suppressor gene dlg is encoding for 21 transcripts. The construct UAS-RNAi-dlg inactivates 14 transcripts--RE, RH, RQ, RS, RG, RD, RL, RB, RK, RR, RT, RN, RA, RP--and does not silenced the other 7 (RO, RF, RI, RU, RJ, RC, RM). This permits to study functions of proteins containg guanilate-kinase domain IPR008145 at C-end of the protein. The most important consequences of the silencing are abnormal mitotic exit and the formation of binuclear cells. Quantitative fluorescence measurements of anti-H3-p histone and DAPI signals showed phase-specific changes in nuclear texture. The inactivation of cellular cortex polarization is the most likely target of dlg inactivation in mitosis.

[Immunogenetic predisposition to chronic nonspecific inflammatory bowel disease].

Immunology has grown beyond the classic doctrine of immunity to infectious diseases, and gradually covered the problems of general physiology and pathology, genetics, embryology, transplantation, oncology and many other disciplines. A new direction has appeared--immunogenetics, which should help to answer questions the disposition and/or resistance to disease, as well as influence of environmental factors on implementation of predisposition to the development of pathology. Much attention is paid to investigation of HLA in IBD. These data indicate a significant polymorphism of major histocompatibility complex antigens in this disease in different countries. The aim of our study was to investigate the immunogenetic susceptibility and resistance to the development of ulcerative colitis (UC), and Crohn's disease (CD), the character of their flow, as well as the associated extraintestinal manifestations, in particular predisposition development of bronchial obstruction (BO) in patients with inflammatory bowel disease (IBD). A study of DNA frozen blood samples of 75 patients with IBD of both sexes has been conducted. The obtained results have been compared with the results of the study 1700 of samples of umbilical cord blood of newborns (apparently healthy children), born at 37-41 weeks' gestation in Moscow (control). The group of patients with UC revealed a positive association of HLA specificities-B*38, HLA-Cw*12 and HLA-DRV1*15, which can be regarded as potentially high risk of developing the disease. The presence of the specificity of HLA-DQV1*02 can be considered as a factor in resistance to the development of UC. High risk of developing Crohn's disease among residents of Moscow associated with groups of alleles of HLA B*41, HLA-B*56, HLA-Cw*05, HLA-Cw*08, HLA-DRV1*01, HLA-DRV1*11, HLA-DQV1*04, and the presence of specificity of HLA-DQV1*05 can be considered as a factor of resistance to the development of BC. High risk of developing BO in patients with IBD is associated with specificities HLA-DQB1*02, DQB1*03, DRB1*15.

[Importance of the immunogenetic factors in the development of glutensensitive celiac disease in adults of Moscow region].

The study examined the genetic predisposition to the development of glutensensitive celiac disease (CD). Were analyzed samples of peripheral blood of 17 adult patients diagnosed with CD, as well as samples of umbilical cord blood of 1700 newborns of healthy children. HLA-typing was performed using two basic methods of molecular typing--SSO and SSP. The studies revealed the specificity of HLA-B*08; HLA-DRB1*2003; HLA-DRB1*07 and HLA-DQB1*02, which are the genetic markers of CD. Was identified a combination of HLA-markers of CD, each of which is a genetic marker for CD: HLA-B*08; HLA-DRB1*2003; HLA-DQB1*02 and HLA-DRB1*2007; HLA-DQB1*02/HLADQB1*03. In addition, negative associations were identified with CD specificity of HLA-DQB1*05. Thus, the results indicate the possibility of individual risk predicting of CD developing in a healthy population and in families of patients with CD.

[Fluorometric identification of chromatin packing level laying out of the light microscopy resolution].

Confocal microscopy permits to perform quantitative analysis of the fluorescent signals of the nuclei. We determined the level of DAPI and phosphorylated histone-H3 fluorescence, the volume of DAPI and phosphorylated histone-H3 fluorescence in normal (Hikone AW) and colchicine treated third instar Drosophila melanogaster wing imaginal disc mitotic cells. Our analysis permitted to indentify two unknown levels of chromatin package; one in the prometaphase and another at the end of metaphase. These levels disappeared in colchicine treated mitoses. We conclude that quantitative analysis of confocal images is able to detect the differences in chromatin package laying over the resolution of light microscopy.

[Nuclear texture in mitotic cells].

Changes of nuclear texture in mitotic cells of Drosophila melanogaster imaginal discs were studied. The distribution of voxels DAPI fluorescence intensities was used as the quantitative measure of the nuclear texture. The integral characteristics such as the portion of voxels with a given fluorescent signal level and autocorrelation of pixel intensities were used. We showed the nuclear texture has specific changes at different mitotic stages and this can be used for more precise staging of mitosis. Colchicines treatment pathologies, connected to abnormal mitoses, by nuclear-texture approach.

[Transplantation of the bone marrow from a HLA-compatible unrelated donor after immunoablative conditioning in children with acquired aplastic anemia unresponsive to combined immunosuppressive therapy: preliminary results].

AIM: To analyze the efficiency of transplantation of the bone marrow from a HLA-compatible unrelated donor and continued immunosuppressive therapy (IST) in children with aplastic anemia (AA) unresponsive to 2 courses of IST. SUBJECTS AND METHODS: The study enrolled 14 children aged 2-16 years (median 9 years). A control group comprised 26 patients in whom IST was continued. The median interval between the diagnosis of AA and transplantation was 26 months (9-156 months). The conditioning regimen consisted of thoracoabdominal irradiation in a dose of 2 Gy, fludarabin (Flu) 100-150 mg/m2, cyclophosphamide (Cy) 100-200 mg/kg, antithymocyte globulin (ATG) in 11 patients and Flu, Cy, and ATG in 3. A graft-versus-host reaction was prevented with mycophenolate mefetil in all the patients, tacrolimus in 11, and cyclosporin A in 3. Donors were compatible for high-resolution typing of 10/10 and 9/10 alleles in 8 and 6 patients, respectively; the source of a transplant was bone marrow in 13 patients and granulocyte colony-stimulating factor-mobilized peripheral blood precursors in one case. RESULTS: Thirteen patients achieved primary engraftment after single transplantation; one patient did after repeat transplantation. Grades I to II graft-versus-host reaction (GVHR) developed in 9 patients; postengraftment life-threatening infections in 3, extensive chronic GVHR in 2, circumscribed GVHR in 7. All fourteen hemopoietic cell transplant recipients followed for a median 17.5 months (range 1-71 months) were survivors. CONCLUSION: The likelihood of good survival after unrelated transplantations in AA is much higher than that after continued IST: 100% versus 15 +/- 11%.

[Dynamics of the spatial organization of the chromosome set in cells of Drosophila melanogaster imaginal disks normally and under the action of the tumor-inducing mutation Merlin].

Fluorescence of H3-p histone and DAPI was studied at different stages of interphase and mitosis in cells of imaginal disks of third-instar Drosophila melanogaster larvae. Three stages differing in the spatial organization of the chromosome set in mitosis were revealed. At the first stage (prophase, prometaphase), the histone 3 phosphorylation level rises, and the volume occupied by the chromosome set in the nucleus increases. The distinctive features of the second stage (metaphase) are a gradual decrease in the histone 3 phosphorylation (the density ofphosphorylation remaining constant) and a reduction of the volume occupied by the chromosome set. At the third stage (anaphase, telophase), the intensity and density of the signal from H3-p histone decrease, and the volume occupied by the chromosome set reduces. At this stage, in Mer4 larvae, in contrast to the control strain, the cells prematurely pass from anaphase into telophase. In addition, a subpopulation of cells with an abnormally large volume of nuclear DNA during the G1 period was revealed in Mer4 larvae. The cells of this subpopulation do not enter into the DNA synthesis and quit the cycle.

[Specifics of anaphase chromatid disjunction in Drosophila melanogaster mitotic mutants].

Anaphase chromatid behavior defects (CBDs) were quantitatively and qualitatively studied in nerve ganglion cells of third-instar larvae of several control wild-type Drosophila melanogaster strains and four strains with mutations of the aar(v158), ff3, mast(v40), and CycB(2g) cell-cycle genes. A linear specificity was observed for the CBD frequency, type, determination, and correction probability. The probability of anaphase CBD correction was close to unity in the control strains and lower in the mutant strains. The lower correction probability in the mutant strains was explained in the context of two findings, that the mutations induced the CBDs that were atypical of the wild-type strains and were potentially uncorrectable in anaphase and that the mutations negatively affected the relative anaphase time in mitosis.