PARP1 is a Versatile Factor in the Regulation of mRNA Stability and Decay

PARP1 is an abundant nuclear protein with many pleiotropic functions involved in epigenetic and transcriptional controls. Abundance of mRNA depends on the balance between synthesis and decay of a particular transcript. PARP1 binds RNA and its depletion results in increased expression of genes involved in nonsense-mediated decay, suggesting that PARP1 might be involved in mRNA stability. This is of interest considering RNA binding proteins play key roles in post-transcriptional processes in all eukaryotes. We tested the direct impact of PARP1 and PARylation on mRNA stability and decay. By measuring the half-lives of two PARP1-mRNA targets we found that the half-lives were significantly decreased in PARP1-depleted cells. PARP1 depletion impacted both the synthesis of nascent mRNA and the stability of mature mRNAs. PARylation impacted the production of nascent mRNA and the stability of mature mRNA, albeit to a lesser extent than PARP1 KD. PARylation enhanced the impact of PARP1 depletion. These studies provide the first direct comparative role of PARP1 and PARylation in RNA stability and decay, adding a new dimension as to how PARP1 regulates gene expression. These studies present a platform to begin to tease out the influence of PARP1 at each step of RNA biogenesis and decay to fine-tune gene expression

Linking Kindling to Increased Glutamate Release in the Dentate Gyrus of the Hippocampus Through the STXBP5/tomosyn-1 Gene

Introduction: In kindling, repeated electrical stimulation of certain brain areas causes progressive and permanent intensification of epileptiform activity resulting in generalized seizures. We focused on the role(s) of glutamate and a negative regulator of glutamate release, STXBP5/tomosyn-1, in kindling. Methods: Stimulating electrodes were implanted in the amygdala and progression to two successive Racine stage 5 seizures was measured in wild-type and STXBP5/tomosyn-1−/− (Tom−/−) animals. Glutamate release measurements were performed in distinct brain regions using a glutamate-selective microelectrode array (MEA). Results: Naïve Tom−/− mice had significant increases in KCl-evoked glutamate release compared to naïve wild type as measured by MEA of presynaptic release in the hippocampal dentate gyrus (DG). Kindling progression was considerably accelerated in Tom−/− mice, requiring fewer stimuli to reach a fully kindled state. Following full kindling, MEA measurements of both kindled Tom+/+ and Tom−/− mice showed significant increases in KCl-evoked and spontaneous glutamate release in the DG, indicating a correlation with the fully kindled state independent of genotype. Resting glutamate levels in all hippocampal subregions were significantly lower in the kindled Tom−/−mice, suggesting possible changes in basal control of glutamate circuitry in the kindled Tom−/−mice. Conclusions: Our studies demonstrate that increased glutamate release in the hippocampal DG correlates with acceleration of the kindling process. Although STXBP5/tomosyn-1 loss increased evoked glutamate release in naïve animals contributing to their prokindling phenotype, the kindling process can override any attenuating effect of STXBP5/tomosyn-1. Loss of this “braking” effect of STXBP5/tomosyn-1 on kindling progression may set in motion an alternative but ultimately equally ineffective compensatory response, detected here as reduced basal glutamate release

STUDENT SOCIO-PSYCHOLOGICAL EXPECTATIONS FROM STUDYING IN HIGHER EDUCATION INSTITUTION: SATISFACTION DEGREE AND IMPLEMENTATION EVALUATION

O objetivo do estudo é analisar a implementação das expectativas dos alunos de estudar na Universidade. O método principal para o estudo desse problema é o método do questionário, que permite o estudo mais qualitativo e aprofundado das expectativas dos jovens em relação ao ensino superior. É revelado que os problemas existentes têm um impacto negativo na atitude dos alunos em relação à sua instituição educacional e, consequentemente, reduzir sua imagem aos olhos dos futuros alunos e do público em geral. Os dados obtidos neste trabalho podem ser utilizados nas áreas de psicologia da educação, psicologia da idade, psicologia social, pedagogia, sociologia, bem como para o desenvolvimento teórico adicional desta questão.El objetivo del estudio es analizar la implementación de las expectativas de los estudiantes al estudiar en la Universidad. El método principal para el estudio de este problema es el método del cuestionario, que permite el estudio más cualitativo y profundo de las expectativas de los jóvenes en relación con la educación superior. Se revela que los problemas existentes tienen un impacto negativo. sobre la actitud de los estudiantes hacia su institución educativa y, en consecuencia, reducir su imagen ante los futuros estudiantes y el público en general. Los datos obtenidos en este trabajo pueden utilizarse en psicología de la educación, psicología de la edad, psicología social, pedagogía, sociología, así como para un mayor desarrollo teórico de este tema.The aim of the study is to analyze the implementation of students' expectations from studying at the University. The leading method for the study of this problem is the method of the questionnaire, which allows the most qualitative and in-depth study of the expectations of young people in relation to higher education.. It is revealed that the existing problems have a negative impact on the attitude of students to their educational institution and, consequently, reduce its image in the eyes of future students and the public as a whole. The data obtained in this work can be used in the psychology of education, age psychology, social psychology, pedagogy, sociology, as well as for further theoretical development of this issue

Personalizing the Way of Teaching LSP

Introduction. The problem of effective teaching language for special purposes (LSP) to the students of non-linguistic professions has been discussed for a long time and in full details by academicians both in Russia and foreign countries. As the result of multiple and thorough surveys general problems of the given situation have been revealed, the way out being defined as special training/retraining of LSP teachers. But in language education publications it is hardly possible to find the description of the program aiming at the training of the above mentioned teachers. The aim of the article is to present effective approaches to teaching LSP and offer the developed master program “Methods of teaching LSP” as a means to solve the problems mentioned above. Materials and Methods. The research methods used are a combination of analytical review of studies by Russian and foreign researchers and interpretation of the results obtained by interviewing university teachers specializing in teaching foreign languages to students completing Bachelorʼs or Masterʼs degree programs. The survey was conducted to assess the level of readiness of professional educators to design the structure and content of programs for teaching LSP, and actually conduct classes within the specified language-teaching area, 81 teachers being responders. Results. As a result of the undertaken review of theoretical works and analysis of the conducted survey a master program “Methods of teaching LSP” was designed for training graduates who will then teach English or Russian as a foreign language at industry-specific higher schools. The developed program focuses on instructing future LSP educators to work out a personalized educational route adjusted to the level of foreign language skills of a student and their prospective professional activities. Discussion and Conclusion. The master program developed by the authors as well as the content of the subjects suggested as its integral part are expected to become a productive way of training LSP educators who are supposed to acquire a number of special skills and competences described in the article. Distant learning and e-learning technologies are also engaged in the program to facilitate learners’ more efficient independent activities. The master program is seen as one of the means of modernizing LSP teachers’ professional training which meets the long-term needs of the Russian education system

Comparison of American mink embryonic stem and induced pluripotent stem cell transcriptomes

BACKGROUND: Recently fibroblasts of many mammalian species have been reprogrammed to pluripotent state using overexpression of several transcription factors. This technology allows production of induced pluripotent stem (iPS) cells with properties similar to embryonic stem (ES) cells. The completeness of reprogramming process is well studied in such species as mouse and human but there is not enough data on other species. We produced American mink (Neovison vison) ES and iPS cells and compared these cells using transcriptome analysis. RESULTS: We report the generation of 10 mink ES and 22 iPS cell lines. The majority of the analyzed cell lines had normal diploid chromosome number. The only ES cell line with XX chromosome set had both X-chromosomes in active state that is characteristic of pluripotent cells. The pluripotency of ES and iPS cell lines was confirmed by formation of teratomas with cell types representing all three germ layers. Transcriptome analysis of mink embryonic fibroblasts (EF), two ES and two iPS cell lines allowed us to identify 11831 assembled contigs which were annotated. These led to a number of 6891 unique genes. Of these 3201 were differentially expressed between mink EF and ES cells. We analyzed expression levels of these genes in iPS cell lines. This allowed us to show that 80% of genes were correctly reprogrammed in iPS cells, whereas approximately 6% had an intermediate expression pattern, about 7% were not reprogrammed and about 5% had a "novel" expression pattern. We observed expression of pluripotency marker genes such as Oct4, Sox2 and Rex1 in ES and iPS cell lines with notable exception of Nanog. CONCLUSIONS: We had produced and characterized American mink ES and iPS cells. These cells were pluripotent by a number of criteria and iPS cells exhibited effective reprogramming. Interestingly, we had showed lack of Nanog expression and consider it as a species-specific feature

Lignin-containing feedstock hydrogenolysis for biofuel component production

In this paper, the commercial 5%Pd/C and 5%Pt/C catalysts and synthesized 5%Pt/MN-270 and 5%Pd/MN-270 were used in the hydrogenolysis of lignocellulosic material (softwood sawdust) to obtain liquid fuels in the form of hydrocarbons. As lignin has a very complex structure, anisole was used as a model compound. It was found that the use Pt-containing catalysts based on hypercrosslinked polystyrene in both processes of anisole and lignin-containing feedstock conversion allowed obtaining the highest yield of oxygen-free hydrocarbons (up to 96 wt. %). Besides, the polymer based catalysts showed high stability in hydrogenolysis process in comparison with the commercial carbon based catalysts

PARP1′s Involvement in RNA Polymerase II Elongation: Pausing and Releasing Regulation through the Integrator and Super Elongation Complex

RNA polymerase elongation along the gene body is tightly regulated to ensure proper transcription and alternative splicing events. Understanding the mechanism and factors critical in regulating the rate of RNA polymerase II elongation and processivity is clearly important. Recently we showed that PARP1, a well-known DNA repair protein, when bound to chromatin, regulates RNA polymerase II elongation. However, the mechanism by which it does so is not known. In the current study, we aimed to tease out how PARP1 regulates RNAPII elongation. We show, both in vivo and in vitro, that PARP1 binds directly to the Integrator subunit 3 (IntS3), a member of the elongation Integrator complex. The association between the two proteins is mediated via the C-terminal domain of PARP1 to the C-terminal domain of IntS3. Interestingly, the occupancy of IntS3 along two PARP1 target genes mimicked that of PARP1, suggesting a role in its recruitment/assembly of elongation factors. Indeed, the knockdown of PARP1 resulted in differential chromatin association and gene occupancy of IntS3 and other key elongation factors. Most of these PARP1-mediated effects were due to the physical presence of PARP1 rather than its PARylation activity. These studies argue that PARP1 controls the progressive RNAPII elongation complexes. In summary, we present a platform to begin to decipher PARP1′s role in recruiting/scaffolding elongation factors along the gene body regions during RNA polymerase II elongation and gene regulation

Vimentin is involved in regulation of mitochondrial motility and membrane potential by Rac1

In this study we show that binding of mitochondria to vimentin intermediate filaments (VIF) is regulated by GTPase Rac1. The activation of Rac1 leads to a redoubling of mitochondrial motility in murine fibroblasts. Using double-mutants Rac1(G12V, F37L) and Rac1(G12V, Y40H) that are capable to activate different effectors of Rac1, we show that mitochondrial movements are regulated through PAK1 kinase. The involvement of PAK1 kinase is also confirmed by the fact that expression of its auto inhibitory domain (PID) blocks the effect of activated Rac1 on mitochondrial motility. The observed effect of Rac1 and PAK1 kinase on mitochondria depends on phosphorylation of the Ser-55 of vimentin. Besides the effect on motility Rac1 activation also decreases the mitochondrial membrane potential (MMP) which is detected by ∼20% drop of the fluorescence intensity of mitochondria stained with the potential sensitive dye TMRM. One of important consequences of the discovered regulation of MMP by Rac1 and PAK1 is a spatial differentiation of mitochondria in polarized fibroblasts: at the front of the cell they are less energized (by ∼25%) than at the rear part

Project Work Management Addressing the Needs of BVI Learners of EFL

Introduction: it is known that inclusive education that opens access to education and facilitates students with special educational needs to socialize successfully is concentrated in special schools whilst mainstream teachers generally lack the methodology of teaching disabled students inclusively. The aim of the article is searching the best ways of educating students with special needs and providing specific conditions of the educational process in mainstream schools. Materials and Methods: the research is based on the results of fulfillment of a number of projects that deal with teaching and educating blind and visually impaired learners and in which all the authors of the article were involved. The problematic approach, the logical method and observation were applied to the research; analysis, comparison, generalization being the most important components of the study. The quantitative data were collected, analyzed and resumed as well. Results: the impact of project work implementation on academic and social skills development of blind and visually impaired students is shown and various types of projects that can be made by visually impaired learners are presented. The authors give their comments when analyzing different ways of adapting research activities to the needs of BVI learners. When speaking in detail on the role of web tools as maximizers of students’ potential the authors refer to their own experience in implementing web tools in project work with blind and visually impaired learners. Discussion and Conclusions: university teacher-training programs should be revised and improved by offering additional courses aimed at educating impaired students through doing research or project work. Special attention should be paid to developing the instructors’ project management skills in terms of teaching English as a foreign language to learners with special needs. It should be noted that different methods and various technological tools are to be used in order to develop research and creative skills of blind and visually impaired learners in an EFL classroom. The article itself, its references and links can be used within the course of lectures and seminars on Methodology of Teaching Foreign Languages, and can also be applied when developing a course of re-training of in-service teachers with particular focus on teaching foreign languages to students (pupils) with special educational needs