Risks in Activities of Educational Institutions: Organizational and Legal Aspects

The article analyses national standards of legal regulation and quality standards in the field of education. The features of using the category of “risk” in relation to the activities of educational institutions in the Russian Federation are covered. Attention is drawn to the most in-demand ways of using the risk-based approach when carrying out state control in various sectors of the economy. The importance of using such methods in education is stressed. Based on the analysis of the dynamics of lawmaking in the Russian Federation the forecast for the development of Supervisory activities in the field of education is made. It is assumed that in the field of education quality management at the state and institutional levels assessment methods and risk management will be needed. It is concluded that the application of a risk-based approach in modern conditions becomes necessary in the activities of the internal system of assurance of higher education institutions of the Russian Federation quality. Scientific novelty and practical significance of the research is seen in what the authors described the basic requirements for the application of risk management in the educational organization. The need for the speedy formation of scientifically grounded methodological basis of risk assessment and management and regulatory underpinning its key provisions are noted

Imitation of optical coherence tomography images by wave Monte Carlo-based approach implemented with the Leontovich–Fock equation

We present a computational modeling approach for imitation of the time-domain optical coherence tomography (OCT) images of biotissues. The developed modeling technique is based on the implementation of the Leontovich–Fock equation into the wave Monte Carlo (MC) method. We discuss the benefits of the developed computational model in comparison to the conventional MC method based on the modeling of OCT images of a nevus. The developed model takes into account diffraction on bulk-absorbing microstructures and allows consideration of the influence of the amplitude–phase profile of the wave beam on the quality of the OCT images. The selection of optical parameters of modeling medium, used for simulation of optical radiation propagation in biotissues, is based on the results obtained experimentally by OCT. The developed computational model can be used for imitation of the light waves propagation both in time-domain and spectral-domain OCT approaches

EFFECT OF DNA CONSTRUCTIONS ELECTROPORATION ON DENDRITIC CELLS

Today transfection of mammalian cell with DNA or RNA construction is the only method for delivering programmed information into the cell nucleus. Electroporation is most commonly used method of transfection in experiments with dendritic cell. The aim of electroporation is to permeabilize the membrane by passing electric impulse through the cell. Due to the increase permeability of the membrane chance DNA or RNA construction getting inside into the cell is increased, wherein survival of the cells is decreased.In the study male mice C57Bl/6 line 2-4 months old were used. From femur bones was isolated 20 × 106 bone marrow cells, which were cultured in 20 mL of complete RPMI-1640 for 7 days. To generate dendritic cells from BM cells, 100 ng/mL of rmFlt3-L was added to culture media at day 0. After 7 days of cultivation, the cell cultures were electroporated with control noncoding plasmids p5 (EP P5) or pmaxCCR9 encoding mouse chemokine receptor CCR9 (EP CCR9). The controls were cell cultures electroporated without any plasmids (mock EP) and cell cultures without electroporation (none EP). 5 × 105 cells were electroporated and resting for 10 minutes. After 10 minutes, cells were harvested and seeded into 24-well plates in 1 mL of culture medium and conditioning medium (1:1). Then, 50 ng/mL of Flt3-L was added to each well. The next day, transfected cells were collected and used for flow cytometry, qRT-PCR analysis.It was found that after electroporation in the groups mock EP, EP P5, EP CCR9 relative amount of live CD11c+ dendritic cells was significantly less than in the non EP group. Moreover, in the EP P5 and EP CCR9 groups the frequency of live CD11c+ dendritic cells was significantly less than in the mock EP group. Expression of the CD86 marker in the EP P5 and EP CCR9 groups was significantly higher than in the non EP and mock EP groups. Expression of the I-AB(MHCII) in the EP CCR9 group on cDC2s was significantly higher than in the non EP group. On plasmacytoid DCs (pDCs) and conventional type 2 DCs (cDC2s) in the EP CCR9 group expression of CCR9 was significantly higher than in the non EP group. Therefore, in this study, we demonstrated the effectiveness of electroporation, accompanied by the decrease in the survival rate and maturation of DCs

Adenine and guanine recognition of stop codon is mediated by different N domain conformations of translation termination factor eRF1

Positioning of release factor eRF1 toward adenines and the ribose-phosphate backbone of the UAAA stop signal in the ribosomal decoding site was studied using messenger RNA (mRNA) analogs containing stop signal UAA/UAAA and a photoactivatable cross-linker at definite locations. The human eRF1 peptides cross-linked to these analogs were identified. Cross-linkers on the adenines at the 2nd, 3rd or 4th position modified eRF1 near the conserved YxCxxxF loop (positions 125–131 in the N domain), but cross-linker at the 4th position mainly modified the tripeptide 26-AAR-28. This tripeptide cross-linked also with derivatized 3′-phosphate of UAA, while the same cross-linker at the 3′-phosphate of UAAA modified both the 26–28 and 67–73 fragments. A comparison of the results with those obtained earlier with mRNA analogs bearing a similar cross-linker at the guanines indicates that positioning of eRF1 toward adenines and guanines of stop signals in the 80S termination complex is different. Molecular modeling of eRF1 in the 80S termination complex showed that eRF1 fragments neighboring guanines and adenines of stop signals are compatible with different N domain conformations of eRF1. These conformations vary by positioning of stop signal purines toward the universally conserved dipeptide 31-GT-32, which neighbors guanines but is oriented more distantly from adenines

МЕТАБОЛИЧЕСКИЕ ПРОЦЕССЫ В ЛИМФОЦИТАХ БОЛЬНЫХ ПРИ ВЕТРЯНОЙ ОСПЕ

Measurement of metabolic processes in lymphocytes in Varicella zoster infection showed highly increased intercellular glycolisys activity with functional cellular overload. Same time, we discovered decreased level of intensivity of first stages of TCC, that rules to lower cycle energetic efficiency and intense metabolitsaminoacides intake for TCC, guiding to high aminoacides transport to lymphocytes. Usage of succinic acid and its salts gives more substrates for TCC, increasing its energetic efficiency and lymphocytes functional activity.Оценка характера и интенсивности метаболических процессов в лимфоцитах больных ветряной оспой по изменениям активности внутриклеточных ферментов показала, что в разгар заболевания отмечена интенсификация реакций гликолиза при значительном повышении функциональной нагрузки на клетки, происходит значительное снижение интенсивности реакций начального этапа ЦТК, что должно уменьшать энергетическую эффективность цикла, а также интенсивное поступление метаболитов на снабжение ЦТК субстратами с аминокислотного обмена, обеспечивая повышенный транспорт аминокислот в лимфоциты. Применение препаратов янтарной кислоты или ее солей дает дополнительное субстратное насыщение ЦТК, повышая энергоэффективность цикла и функциональные возможности лимфоцитов

Statistical Analysis of Readthrough Levels for Nonsense Mutations in Mammalian Cells Reveals a Major Determinant of Response to Gentamicin

The efficiency of translation termination depends on the nature of the stop codon and the surrounding nucleotides. Some molecules, such as aminoglycoside antibiotics (gentamicin), decrease termination efficiency and are currently being evaluated for diseases caused by premature termination codons. However, the readthrough response to treatment is highly variable and little is known about the rules governing readthrough level and response to aminoglycosides. In this study, we carried out in-depth statistical analysis on a very large set of nonsense mutations to decipher the elements of nucleotide context responsible for modulating readthrough levels and gentamicin response. We quantified readthrough for 66 sequences containing a stop codon, in the presence and absence of gentamicin, in cultured mammalian cells. We demonstrated that the efficiency of readthrough after treatment is determined by the complex interplay between the stop codon and a larger sequence context. There was a strong positive correlation between basal and induced readthrough levels, and a weak negative correlation between basal readthrough level and gentamicin response (i.e. the factor of increase from basal to induced readthrough levels). The identity of the stop codon did not affect the response to gentamicin treatment. In agreement with a previous report, we confirm that the presence of a cytosine in +4 position promotes higher basal and gentamicin-induced readthrough than other nucleotides. We highlight for the first time that the presence of a uracil residue immediately upstream from the stop codon is a major determinant of the response to gentamicin. Moreover, this effect was mediated by the nucleotide itself, rather than by the amino-acid or tRNA corresponding to the −1 codon. Finally, we point out that a uracil at this position associated with a cytosine at +4 results in an optimal gentamicin-induced readthrough, which is the therapeutically relevant variable

Far upstream element binding protein 1 binds the internal ribosomal entry site of enterovirus 71 and enhances viral translation and viral growth

Enterovirus 71 (EV71) is associated with severe neurological disorders in children, and has been implicated as the infectious agent in several large-scale outbreaks with mortalities. Upon infection, the viral RNA is translated in a cap-independent manner to yield a large polyprotein precursor. This mechanism relies on the presence of an internal ribosome entry site (IRES) element within the 5′-untranslated region. Virus–host interactions in EV71-infected cells are crucial in assisting this process. We identified a novel positive IRES trans-acting factor, far upstream element binding protein 1 (FBP1). Using binding assays, we mapped the RNA determinants within the EV71 IRES responsible for FBP1 binding and mapped the protein domains involved in this interaction. We also demonstrated that during EV71 infection, the nuclear protein FBP1 is enriched in cytoplasm where viral replication occurs. Moreover, we showed that FBP1 acts as a positive regulator of EV71 replication by competing with negative ITAF for EV71 IRES binding. These new findings may provide a route to new anti-viral therapy