Differentiation between <I>Yersinia pestis</I> Strains of Altaic-Hissar Group of Non-Main Subspecies by Means of PCR

On the basis of comparison between the genome-wide sequences of Altaic and Hissar strains of non-main subspecies and strains from Talas high-mountain plague focus, identified have been two novel DNA-targets, the usage of which provides for easy rapid subdivision of these closely-related strains of non-main subspecies that fall into separate phylogenetic brunches of plague agent evolution, applying PCR assay. One of the targets is allocated to intergenic region between YPO3333 and YPO3332 genes. In strains of altaica ssp. it contains 122 bps deletion. The other target is YPO2412 gene region which contains 72 bps deletion in Talas strains. Special primers for DNA-targets have been designed. Established have also been test specifications. Efficacy of the method for differentiation between the strains of Altaic-Hissar group of non-main subspecies is validated on 97 Y. pestis strains of various ssp

Development of a Set of Primers for Drug-Resistance Genes Detection in the Agents of Dangerous Bacterial Infections as Exemplified by <I>Yersinia pestis</I>, <I>Vibrio cholerae</I>, <I>Escherichia coli</I> Strains

A set of primers for detection of genes encoding resistance to streptomycin ( strA, strB ), tetracyclin ( tetA, tetR ), chloramphenicol ( catА ), kanamycin ( npt , aphA ), vankomycin ( sanA ), polymyxin ( pmrD ) has been developed with the aim of rapid and effective detection of drug-resistant strains of dangerous bacterial infections agents. Efficacy of constructed primers has been confirmed against a panel of 40 Yersinia pestis, 49 Vibrio cholerae, and 2 Escherichia coli strains from the State collection of pathogenic bacteria of the RAPI “Microbe”. Drug-resistance genes ntp and catA have been detected in plague agent strains , strA, strB , npt , aphA , tetA and tetR - in cholera agent; strA , tetR , ntp and aphA - in pathogenic strain E. coli О157:H7. Determined is universal character of the designed primers for drug-resistance genes detection in these pathogenic bacteria species

BTK, NuTM2A, and PRPF19 are Novel KMT2A Partner Genes in Childhood Acute Leukemia

Chromosomal rearrangements of the human KMT2A/MLL gene are associated with acute leukemias, especially in infants. KMT2A is rearranged with a big variety of partner genes and in multiple breakpoint locations. Detection of all types of KMT2A rearrangements is an essential part of acute leukemia initial diagnostics and follow-up, as it has a strong impact on the patients’ outcome. Due to their high heterogeneity, KMT2A rearrangements are most effectively uncovered by next-generation sequencing (NGS), which, however, requires a thorough prescreening by cytogenetics. Here, we aimed to characterize uncommon KMT2A rearrangements in childhood acute leukemia by conventional karyotyping, FISH, and targeted NGS on both DNA and RNA level with subse-quent validation. As a result of this comprehensive approach, three novel KMT2A rearrangements were discovered: ins(X;11)(q26;q13q25)/KMT2A-BTK, t(10;11)(q22;q23.3)/KMT2A-NUTM2A, and inv(11)(q12.2q23.3)/KMT2A-PRPF19. These novel KMT2A-chimeric genes expand our knowledge of the mechanisms of KMT2A-associated leukemogenesis and allow tracing the dynamics of minimal residual disease in the given patients. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Funding: KMT2A rearrangement assessment was supported by the Russian Science Foundation (grant no. 19-75-10056). Quantitative RT-PCR for MRD monitoring was supported by Russian Presidential (grant no. MK-1645.2020.7)

Identification and Intraspecific Typing of Plague Microbe Strains with Their Potential Virulence Determination Using PCR

Developed is the method of identification and intraspecific typing of plague microbe strains along with their potential virulence determination. Intraspecific differentiation and focal attribution of the examined plague microbe strains can be determined by monolocus VNTR-PCR, and main virulence determinants (chromosomal pigmentation region and calcium-dependence plasmid genes) - by multiplex PCR

Exploratory Study of the Long-Term Persistence of <i>Yersinia pestis</i> in the Cells of Soil-Inhabiting Ameba - <i>Acanthamoeba Sp.</i>

Objective of the study is to explore the feasibility of the long-term persistence of Yersinia pestis strains in association with ameba - Acanthamoeba sp. Materials and methods. Investigated has been interaction of ameba - Acanthamoeba sp., isolated from rodent burrows in the Pre-Caspian sandy, Volga-Ural steppe, and Pre-Caspian North-Western steppe natural foci, with 4 strains of Y. pestis of the main subspecies, 1 strain of caucasica and 1 strain of altai subspecies. Results and discussion. It is established that the strains of the main subspecies survive in the cells of ameba at 26 °C and 20 % humidity (modeling of the drought conditions in the natural plague foci) within 2-4 months of experiment, and 10-20 times longer that in pure culture. Two strains of the non-main ssp. have not demonstrated an increase in persistency in association with Acanthamoeba sp., which may occur due to degraded resistance to phagocytosis in the ameba of this specie. Using fluorescent and transmission electronic microscopy, it is determined that the cells of plague agent persist in ameba cells in individual vacuoles, enclosed in endoplasmic reticulum. The data obtained may testify to the possible involvement of ameba Acanthamoeba sp. . into sustainment of Y. pestis in soil biocoenoses of natural plague foci

Hematopoietic stem cell transplantation with alpha/beta T-lymphocyte depletion and short course of eculizumab in adolescents and young adults with paroxysmal nocturnal hemoglobinuria

The main goal is to optimize hematopoietic stem cell transplantation (HSCT) approach among adolescents and young adults with paroxysmal nocturnal hemoglobinuria (PNH) by means of Graft-versus-host disease (GVHD) and post-transplant complications risk lowering. Materials and methods. We report our experience of HSCT from HLA-matched unrelated donors using TCR alfa/beta and CD19 depletion in 5 pts (1M/4F) with PNH, developed after successful immunosuppressive therapy (IST) of acquired aplastic anemia (AA). Median age of pts at the moment of transplantation was 17,8 years (range 14,5-22,7), median interval from IST to PNH was 4 years (5mo - 6,5 y). In all patients non-severe pancytopenia was present: granulocytes 0,8х109/l (0,8-1,8 х109/l) platelets 106 х109/l (27-143 х109/l) and Hb -78 g/l, median PNH clone size in granulocytes was 94 (range 75-99)%. One pts previously developed sinus thrombosis. Conditioning consisted of thoraco-abdominal irradiation 4-6 Gy, cyclophosphamide 100 mg/kg, fludarabine 150 mg/m2 and anti-thymocyte globulin (ATG) or alemtuzumab. Eculizumab was given from day (-7) till day (+14) (every 7 days, only 4 times). GVHD prophylaxis was tacrolimus ± methotrexate. Results. Infusedgraft characteristics were: CD34+ - 8,1х106/kg, CD3TCRab·150х103/kg, CD3gd+ - 7,3х106/kg, СD19+ - 221х103/kg, NK -6,4х108/kg. Engraftment was achieved in all 5 pts with a median of 15(12-18) и 13(10-18) days for granulocytes and platelets, respectively. Skin acute GVHD grade I developed in only 1 pt, and subsided with short course of glucocorticoids. CMV reactivation occurred in 1 pt; there were no episodes of Epstein-Barr Virus (EBV) o rAdenovirus (AdV) reactivation. Full donor myeloid chimerism was established in all pts by day +30. Immune reconstitution was delayed until 6 months after transplant but no severe infections occurred. All pts are alive 1,7-5,5 years (med 4 years) after HSCT with normal hematopoiesis and immune function, full donor chimerism and no late sequelae. Conclusions. Transplantation of TCRalfa/beta and CD19 depleted hematopoietic cells from matched unrelated donor after immunoablative conditioning and supported with short course of eculizumab is perfectly safe and efficient technology leading to cure in young patients with PNH

An environment for relation mining over richly annotated corpora: the case of GENIA

BACKGROUND: The biomedical domain is witnessing a rapid growth of the amount of published scientific results, which makes it increasingly difficult to filter the core information. There is a real need for support tools that 'digest' the published results and extract the most important information. RESULTS: We describe and evaluate an environment supporting the extraction of domain-specific relations, such as protein-protein interactions, from a richly-annotated corpus. We use full, deep-linguistic parsing and manually created, versatile patterns, expressing a large set of syntactic alternations, plus semantic ontology information. CONCLUSION: The experiments show that our approach described is capable of delivering high-precision results, while maintaining sufficient levels of recall. The high level of abstraction of the rules used by the system, which are considerably more powerful and versatile than finite-state approaches, allows speedy interactive development and validation

The role of nelarabine in the treatment of T-cell acute lymphoblastic leukemia: literature review and own experience

Aim. The analysis of experience of nelarabine use in refractory/relapsed T-cell acute lymphoblastic leukemia (T-ALL) depending on the immunophenotype and the line of therapy. Materials and methods. All the patients with relapsed or refractory T-ALL aged from 0 to 18 years who received treatment with nelarabine as a part of the therapeutic element R6 were included in the study. For all patients a detailed immunological analysis of leukemia cells with discrimination of immunological variants TI, TII, TIII or TIV was performed. Patients administered with nelarabine as a first therapeutic element were referred to the first-line therapy group, other patients were referred to the second-line therapy group. Nelarabine was administered as intravenous infusion at a dose of 650 mg/m2, on days 1-5. Allogeneic hematopoietic stem cells transplantation (allo-HSCT) was considered for all patients. Results. From 2009 to 2017, 54 patients with refractory/relapsed T-ALL were treated with nelarabine. Five-year event-free survival (EFS) and overall survival (OS) was 28% for all patients, cumulative risk of relapse (CIR) was 27%. EFS was significantly higher in nelarabine first-line therapy group in comparison with second-line therapy group (34±8% vs 8±8%, p=0,05). In patients after allo-HSCT EFS, OS and CIR were 51±10%, 50±10% and 39,1±9,5% accordingly. The best results were achieved in patients with TI immunophenotype. No toxicity-related mortality as well as severe neurologic complications or discontinuation of therapy associated with use of nelarabine were reported. Conclusion. The use of nelarabine is an effective strategy for the treatment of relapsed and refractory T-ALL. The best treatment outcomes were obtained in patients with TI immunophenotype and in the first-line therapy group. Optimal dosage regimens can be established during controlled clinical trials

Studies of Biofilm Formation in Non-Pigmented and Plasmid-Deprived Mutants of <I>Yersinia pestis</I> on Biotic Surfaces, <I>in vivo</I> and <I>in vitro</I> Conditions

In non-pigmented and plasmid-deprived mutants – isogenic variants of highly virulent Yersinia pestis 231 strain – studied is the mechanism of biofilm formation on biotic surfaces, both in vitro (on the laboratory model of nematode Caenorhabdiitis elegans) and in vivo (inside the alimentary tract of Nosopsyllus laeviceps flea). It is determined that spontaneous loss of ability to form biofilms and generate pigmented colonies in the mutants is probably caused not only by the deletion of the whole chromosome pigmentation fragment, but also by a point(single base) mutation in structural hms operon. It is demonstrated that the absence of pCad, pFra or pPst plasmids does not have an impact on the ability of plasmid-deprived mutants to form biofilm on the cuticle of nematode C. elegans

Особенности использования мультимедийных презентаций и электронных учебников при обучении иностранных граждан в российских вузах

The article raises the problem of training cadets of the CIS countries who receive higher education in military universities of our country. Russian language proficiency is the main problem, as the training is exclusively in Russian, and as part of a Russian group. Language acquisition occurs gradually during interpersonal communication, and most of all, at classes. One of the first subjects that cadets start studying is mathematics. The main feature of mathematics is the universality of formulas for communication in its language both in the CIS countries and in the world. It greatly simplifies the process of transmitting information to students, allows the teacher to minimize the number of Russian words at the first stages of training (first year of training) gradually raising the volume of definitions and theorems. The great majority of students from the CIS countries freely perceive the lecture and fully assimilate the information even in such a section of «Higher mathematics»as probability theory by the end of the second year. The article shows the transition from the overwhelming number of formulas in the lectures to the prevalence of definitions and theorems. The number of Russian words also increases at practical classes. Their introduction is based on drawings (topic «Analytical geometry»), graphs («Introduction to mathematical analysis», «Definite integral »). The laboratory works are of the greatest interest, of course, where students must draw conclusions about the suitability of the formula obtained as the experiment result. The article details the methodological features of organizing various types of classes using multimedia tools and electronic textbooks in groups with cadets from the CIS countries.В данной статье поднимается проблема обучения курсантов стран СНГ, которые получают высшее образование в военных вузах нашей страны. Следует отметить, что проблема связана, в первую очередь, с владением русским языком, так как обучение осуществляется исключительно на русском языке и в составе русской группы. Овладение языком происходит постепенно в ходе межличностного общения, а больше всего, во время занятий. Одним из первых предметов, которые начинают изучать курсанты, является математика. Основной особенностью математики можно считать универсальность формул для общения на ее языке не только в странах СНГ, но и во всем мире, что значительно упрощает процесс передачи информации обучающимся, позволяет преподавателю минимизировать количество русских слов на первых этапах обучения (первый год обучения), постепенно увеличивая объем определений и теорем. К концу второго курса курсанты из стран СНГ, в подавляющем большинстве, свободно воспринимают лекцию и полностью усваивают информацию даже по такому разделу дисциплины «Высшая математика», как теория вероятностей. В статье на примерах показан переход от подавляющего количества формул на лекции к превалированию определений и теорем. На практических занятиях также происходит увеличение количества русских слов. Их введение опирается на чертежи (тема «Аналитическая геометрия»), графики («Введение в математический анализ», «Определенный интеграл»). Но, безусловно, наибольший интерес вызывают лабораторные работы, в ходе выполнения которых обучаемые должны сделать выводы о пригодности полученной в результате эксперимента формулы. В статье подробно описаны методические особенности организации различных видов занятий в группах, где присутствуют курсанты стран СНГ, с использованием мультимедийных средств и электронных учебников