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)

SURVIVAL OF BURKHOLDERIA PSEUDOMALLEI IN CELLS OF TETRAHYMENA PYRIFORMIS CILIATE INFUZORIAN: EFFECT ON TETRAHYMENA ENCYSTMENT ACTIVITY

Objective of the study was to model the interaction of Burkholderia pseudomallei with Tetrahymena pyriformis in vitro and investigate the changes in the population composition of the protozoa when co-cultured with a microorganism.Materials and methods. B. pseudomallei 110, C141, 57576, 107 strains differing in virulence for BALB/c mice were used. The axenic culture of T. pyriformis was incubated with microorganisms in 100 to 1 ratio, at 28 °C, in LB. Samples of co-cultures were examined using light microscopy, by counting the number of trophozoites and cysts in the population. Dynamics of multiplication of B. pseudomallei cultures associated with T. pyriformis was determined through seeding bacteria on a dense nutrient medium to count the grown colonies.Results and conclusions. B. pseudomallei in association with T. pyriformis is ingested by protozoan cells; it multiplies in them and stimulates protozoa encystment. Hereby virulent strain B. pseudomallei 110 induces encystment of T. pyriformis on days 2–4 and complete cell destruction within 7–8 days. Avirulent strain, B. pseudomallei 107, induces full encystment on day 7; significant part of the cysts remains intact on day10. Dynamics of B. pseudomallei growth, co-cultured with T. pyriformis is characterized on day 1 by distinct decrease in the number of viable bacterial cells and increase in it within following 24 hours. Bacteria concentration curves depend on the virulence of the strain: maximum level of B. pseudomallei 110 replication is observed after 48 hours, while that of B. pseudomallei 107 – not less than after 7–8 days

Phenotypic diversity of chickpea (Cicer arietinum L.) landraces accumulated in the Vavilov collection from the centers of the crop origin

Chickpea (Cicer arietinum L.) is the second grain legume for the area of cultivation in the world, and the third for the production. However, modern cultivars of chickpea are typically susceptible to a variety of diseases, and have modest drought tolerance. The improvement of the crop for adaptability сould be carried out via introgression of valuable traits genes from old landraces collected in the centers of chickpea origin and diversity: the primary – Turkey and secondary – Ethiopia. The N.I. Vavilov All-Russian Institute of Plant Genetic Resources (thereafter VIR) preserves 3380 chickpea accessions, with landraces representing over a half of them. Here, the results of analysis of variability of 11 biological, morphological and economic-valuable traits in 1082 chickpea landraces descended from 60 countries are briefly driven. More in detail the sample of 75 landraces from Turkey and 24 landraces from Ethiopia (centres of chickpea origin) which had been sampled there 90 years ago have been studied. We analyzed the phenotypic variability with a treatment of 15 traits. The traits were studied using component analyses. Geographic regularities of certain traits in the studied accessions have been revealed. Ethiopian landraces are relatively homogeneous, belonging mostly to desi-type, and having fewer small, dark, and angular seeds, a short maturation period. They belong to the Abyssinian eco-geographical group, absolutely unique and endemic to Ethiopia. Turkish landraces are characterized by much higher diversity for the majority of phenotypes, covering almost the entire range of traits specified in chickpea descriptors. In this region, together with landraces typical for Turkey, there are those from the western Mediterranean and from the areas bordering with Turkey to the east. Landraces from primary and secondary centers of origin differed on the range of variability of the traits studied. The smaller degree of variation and primitiveness of the most traits and lower seed productivity in Ethiopian landraces in comparison with Turkish ones indicates a greater breeding advancement of the latter. Useful traits for breeding are present in the landraces from both centers of origin and diversity

Burkholderia pseudomallei Morphotypes that Form in vitro under Stress Conditions

Objective of the study was to determine diversity of the morphotypes formed in vitro from the initial morphological variant of B. pseudomallei 110 under stressful conditions and to study some phenotypic characteristics of them. Materials and methods. Virulent strain Burkholderia pseudomallei 110 of Australian serotype was used. Burkholderia cultures were added to the axenic culture of Tetrahymena pyriformis in LB broth and sterile river water in the ratio of 100: 1 and incubated at 28 °C; the passage of monocultures and cultures in protozoa cells was repeated at intervals of 3–4 days. Morphotypes were identified on Ashdown’s medium after cultivation for 3–4 days at 32 °C, photographs were analyzed based on classification of Chantratita et al. In all morphotypes the activity of extracellular enzymes and virulence were determined on the model of golden hamsters. Results and conclusions. Seven B. pseudomallei 110 colony morphotypes were identified. Four of them with characteristics of I, III, IV and VII morphotypes, described by Chantratita et al., were named Chl (Chantratita like variant). The study of morphotypes in different samples revealed a variation in them, depending on the culture medium (LB broth or water), and their different ratios in individual samples. The greatest number of morphological variants (4 out of 7) was formed during the passage of the monocultures of B. pseudomallei 110 in LB broth; in water the initial culture was almost entirely (95 %) transformed into morphotype I Chl. Under other conditions of cultivation the dominant V morphotype was formed, and in the presence of protozoa it was combined predominantly with I Chl. Morphotypes differed in the production of extracellular enzymes, motility and reduced virulence

Current State of the Development of Therapies for Emergency Prophylaxis and Treatment of Ebola Virus Disease

Nowadays vaccination of the population living in the endemic regions and widespread implementation of the potent therapies for the emergency prophylaxis and treatment into the clinical practice are regarded as the basic efficient and cost-effective measures for Ebola epidemic spread control. Objective of the review is to analyze current state of the development of aids for the immediate prophylaxis and treatment of Ebola fever. Focus area of the activities is the construction of drugs on the basis of virus-specific anti-bodies (including monoclonal), small interfering RNA, and anti-sense phosphordiamidate morpholine oligomers and interferons. The paper discusses the most significant achievements in this sphere

Positive and negative properties of four endodontic sealant groups: a systematic review

Background. The choice of sealant is an important dentist’s decision with a long-term influence on treatment. Knowledge of the properties and characters of each material is key to the optimal endosealer selection on individual basis.Objectives. A comparison of endodontic sealants based on epoxy resins, calcium hydroxide, zinc oxide eugenol and bioceramics.Methods. Publications were mined in the PubMed and Google Scholar electronic databases to cover the four sealer groups (epoxy, calcium hydroxide, zinc oxide eugenol and bioceramics) over years 2014-2021, including selected relevant sources within 2002-2013; 73 articles were considered for review, regardless of the study design or language. Content and descriptive analyses were used as research tools. Meta-analysis was not used due to a high evidence heterogeneity.Results. The dental market currently offers a variety of sealant groups. Endodontic sealants based on epoxy resins, zinc oxide eugenol, calcium hydroxide and bioceramics are the most common. The review identifies no ideal material for the root canal filling.Conclusion. Manifold studies demonstrate the pros and cons in each endodontic sealant group with respect to variant criteria. Depending on clinical situation, the practitioner can opt for the material property to identify the endosealer

Future Developments and Applications of the Vaccines against Dangerous Viral Infections, RNA-Replicon-Based, Obtained from the Venezuelan Equine Encephalomyelitis Virus

The members of the Filoviridae (Marburg and Ebola viruses) and Arenaviridae (Lassa, Lujo, Machupo, Junin, Guanarito, Sabia viruses) families are the etiological agents of particularly dangerous viral hemorrhagic fevers. These agents pose a potential threat to public health care in view of the possibility of their unintended import into the non-endemic regions, and thus construction of specific medical protectors as regards induced by them diseases is a pressing issue. According to leading experts, vaccination of the cohorts that fall in the risk groups is the most effective and least expensive method to prevent the development of epidemics. The review contains information on a new prospective line of protective preparations development as regards particularly dangerous viral infections - construction of alphavirus-replicon-based vaccine. Elaboration of recombinant replicons does not require cultivation of pathogenic microorganisms. RNA-replicons are distinguished by their incapacity to produce infective progeny, which is of a great importance for the development of vaccines against particularly dangerous viral hemorrhagic fevers. Advantages of alphaviral replicons over other RNA-replicons are as follows: high levels of heterologous gene expression and resistance to anti-vector immunity. RNA-replicons of alphaviruses combine the safety of inactivated, and immunogenicity of live attenuated vaccines. Alphaviruses-based replicons are suitable for express vaccine development with the purpose of specific prophylaxis of viral infectious diseases

The Molecular Genetic Peculiarities of Genomic Structure of Members of the <i>Ebolavirus</i> Genus

The molecular genetic peculiarities of genomic structure of the Ebolavirus genus members are viewed in the review. The Ebola virus disease outbreaks in West African countries constitute a threat not only for Africa, but for the whole world in view of possible introduction of the agent in non-endemic regions. The members of the Ebolavirus genus have different pathogenicity for humans, thus differ severity and mortality of the disease they cause. There is a significant genetic divergence among members of the Ebolavirus genus. The differences of pathogenic potential of members of the Ebolavirus genus may be explained as the result of mutations in the genes of virus structural proteins. It is possible, that some of these mutations may affect virulence of strains within one virus species. So far as most effective modern medicines for specific prophylaxis and treatment of Ebola fever are target-oriented, genotyping of the agent will promote elaboration of strategy of such preparations development

The comparative analysis of systemic inflammatory reaction in ankylosing spondylitis and rheumatoid arthritis

Rheumatoid Arthritis (RA) and ankylosing spondylitis (AS) are chronic inflammatory joint diseases. It was shown, that autoimmune diseases pathogenesis can be considered as typical pathological process chronic systemic inflammation, where systemic inflammatory reaction (SIR) plays the main role. The SIR pathological importance in RA is obvious. SIR in AS is less observed. It determines the expediency of systemic inflammation markers comparative analysis in this diseases. The research objective is to compare SIR expressiveness in AS and RA using integral characteristics. 25 AS patients and 26 RA patients were examined. The control group included 50 healthy donors. Such SIR markers were measured in blood plasma using immunochemilumeniscention method: C-reactive protein (CRP), interleukine (IL)-6, IL-8, IL-10, tumor necrosis factor (TNF) a, mioglobin, troponin I, The integral SIR characteristic reactivity coefficiency (RC) was calculated. The significant elevation of CRP, IL-6, IL-8, TNFa, mioglobin levels was detected in RA and AS groups. The significant IL-6 level elevation was detected in RA group by comparison RA and AS groups. In investigated groups were not found significant distinctions in RC values and elevation frequency. In AS and RA patients SIR is shown in form of acute phase response and hypercytokinemia. The systemic alteration signs are detected only in certain cases. On SIR expressiveness and systemic alteration probability RA and AS can be carried to one autoimmune diseases type.Ревматоидный артрит (PA) и анкилоэрующий спондилит (АС) являются тяжёлыми воспалительными заболеваниями суставов. Показано, что патогенез аутоиммунных заболеваний можно рассматривать с точки зрения типового патологического процесса хронического системного воспаления, ведущую роль в котором играет системная воспалительная реакция (СВР). Патогенетическое значение СВР при РА в настоящее время очевидна. При АС роль СВР расшифрована в меньшей степени, что определяет целесообразность сравнительного анализа проявлений отдельных критериев системного воспаления при этих нозологиях. Цель исследования - сравнить выраженность СВР при АС и РА, используя интегральные показатели. Материалы и методы: Обследовано 25 больных АС и 26 больных РА. Контрольная группа - 50 здоровых доноров. В плазме крови определялись показатели: С-реактивного белка (CRP), интерлейкина (И)-6, IL-8, IL-10, фактора некроза опухоли-а (TNFa), тропонина I, миоглобина методом иммунохемолюменисценции. Рассчитывался интегральный показатель СВР — коэффициент реактивности (КР). Результаты и обсуждение: У больных АС и РА выявлено достоверное увеличение содержания CRP, IL-6, TNFa, IL-8, IL-10, миоглобина относительно лиц контрольной группы. При сравнении показателей СВР в группах РА и АС получены достоверные различия только по уровню IL-6, содержание которого выше в группе РА. В исследуемых группах не найдено достоверных различий по значению и частоте повышения КР. У больных РА и АС СВР проявляется в виде острофазного ответа и гиперцитокинемии. Признаки системной альтерации регистрируются в отдельных случаях. Таким образом, по выраженности СВР и вероятности проявления системной альтерации РА и АС можно отнести к одному классу аутоиммунных заболеваний

Translocation t(1;11)(p32;q23) with MLL-EPS15 fusion gene formation in acute leukemias: a review and 6 new case reports. Approaches to minimal residual disease monitoring

We performed clinical and laboratory characterization of patients with rare translocation t(1;11)(p32;q23) leading to MLL-EPS15 fusion gene formation. Study cohort consisted of 33 primary acute leukemia (AL) cases including 6 newly diagnosed and 27 patients previously described in literature. Among study group patients t(1;11)(p32;q23) was found most frequently in infant AL cases (median age 8 months). In acute lymphoblastic leukemia (ALL) male/female ratio was 1:3, in acute myeloid leukemia (AML) it was 1:1. Additional cytogenetic aberrations in 38 % of patients were revealed. The most frequent breakpoint position in EPS15 gene was intron 1. Four different types of MLLEPS15 fusion gene transcripts were detected. Primers-probe-plasmid combination for MLL-EPS15 fusion gene transcript monitoring by realtime quantitative polymerase chain reaction (RQ-PCR) was developed and successfully applied. In 3 patients RQ-PCR was done on genomic DNA for absolute quantification of MLL-EPS15 fusion gene. High qualitative concordance rate (92 %) was noted between minimal residual disease data obtained in cDNA and genomic DNA for MLL-EPS15 fusion detection.</p