Phenotypic traits differentiating the genetic resources of pea (Pisum sativum L.) by the type of use.

The paper presents an analysis of the data obtained for pea accessions from the VIR collection studied at the Adler Experiment Station in the setting of the Krasnodar Territory in 2017-2019. It was for the f irst time that these accessions were studied for a set of phenotypic traits. The object of the study was a sample of 494 pea accessions originated from 43 countries and 18 regions and territories of the Russian Federation. The work was carried out in compliance with the methodological guidelines developed at VIR. Statistica 13.3 software was employed for statistical data processing. An assessment of four qualitative, 10 quantitative and four phenological traits in the accessions made it possible to differentiate them by the type of use, that is, as dry, forage and garden peas. The varieties differing in the type of use signif icantly differed by the values of such traits as stem length, number of pods per plant, number of nodes to the f irst f lower, number of f lowers in the inf lorescence, the maximum number of seeds per pod, pod length, and a narrower pod of forage pea compared to that of dry and garden peas. The average values of these traits were recorded for the peas with different types of use. The maximum difference was noted between garden and forage pea varieties. Dry pea varieties occupied an intermediate position. The complex of phenotypic traits identif ied determines the differences between three types of pea use, which is important when selecting the initial material for breeding appropriate varieties.

[Herd immunity to SARS-CoV-2 in the Novosibirsk Region population amid the COVID-19 pandemic].

AIM: To determine the level of SARS-CoV-2 seroprevalence among the Novosibirsk Region population against the background of the COVID-19 pandemic. MATERIAL AND METHODS: The work was carried out in 2 phases: 1) a cross-sectional cohort study performed 28.06- 15.07.2020; 2) longitudinal cohort 3-stage seromonitoring: 1st stage 28.06-15.07.2020; 2nd 14.09-04.10.2020; 3rd 10-30.12.2020 The work was carried out according to a unified methodology developed by Rospotrebnadzor with the participation of St-Petersburg Pasteur Institute, taking into account the recommendations of the WHO. IgG antibodies to the SARS-CoV-2 nucleocapsid protein were detected by ELISA using a kit of reagents produced by the SRCMSB (Obolensk) according to the manufacturer's instructions. Statistical analysis was performed using Microsoft Excel 2010 and other programs. RESULTS: The seroprevalence in the region's population was 9.1% (95% CI 8.0-10.2): maximum in children 14-17 years old (17.6%, 95% CI 12.3-23.9) and persons over 75 years (14.8%, 95% CI 11.4-18.8), minimum among persons 30-39 years old (4.9%, 95% CI 3.0-8.0). Increased rate was noted among the unemployed (15.4%, 95% CI 9.9-17.1) and other individuals (13.0%, 95% CI 8.6-18.5). Seroprevalence was 33.3% (95% CI 16.3-59.0) in COVID-19 convalescents and 19.0% (95% CI 13.9-25.0) in contact persons. More than 94.7% (95% CI 91.2-97.2) of seropositive individuals were asymptomatic. During the serological monitoring, seroprevalence increased from 7.4% (95% CI 6.2-8.9) at 1st stage 1 to 12.4% (95% CI 10.6-14.3) at 2nd , and 31% (95% CI 28.8-33.3) at 3rd stage. CONCLUSION: SARS-CoV-2 herd immunity has not reached the threshold level, this does not exclude exacerbation of the epidemic process.

[Modified method for determining the normal values of the aortic root diameter in the focus of aortic pathology in individuals with congenital connective tissue disorders].

INTRODUCTION: Enlargement of the aortic root of occurs in many cases with cardiovascular disease, including congenital connective tissue disorders (CCTD), especially its differentiated and undifferentiated forms (UCCTD). A common deficiency of the generally accepted methods for diagnosing an aortic root dilation may be that persons with CCTD and cardiovascular diseases of potentially having a broader aortic root diameter could be included in the reference groups. The purpose of our study was to develop a modified method for determining the normal aortic root diameter in group of individuals without CCTD and cardiovascular diseases. MATERIALS AND METHODS: The study included 464 apparently healthy people, men and women aged 15 to 65 years. All patients underwent general clinical examination, echocardiography. On the basis of external and internal signs of CCTD, patients with UCCTD were identified - the UCCTD group (n = 208) and without this pathology - the main group (n = 256). The calculation of the normal aortic root diameter (NARD) was made in accordance with the methods of Roman M. J. et al. (1989) and Devereux R. B. et al. (2012). The upper limit values of NARD (UL NARD) were calculated according to the algorithms of Roman M. J. et al. (1989), Devereux R. B. et al. (2012), Campens L. et al. (2014). RESULTS: The data obtained in the main group was used to develop a modified method for the determination of NARD. The mean values by echocardiography and calculated values of the aortic root by modified method of this study were practically the same in the main group, whereas the mean value of NARD calculated by the methods of Roman M. J. et al. and Devereux R. B. et al. in this sample were significantly higher in relation to the listed values. As the NARD values, the UL NARD were significantly higher for all evaluated algorithms in comparison with the modified method. In the group of patients with UCCTD, 13 cases of aortic root dilation were found according to the method of Roman M. J., compared to 19 cases by the modified method. At the same time, 3 patients with aortic root enlargement by the mew method had 7 points of systemic involvement, thus corresponding to the Ghent criteria of Marfan syndrome. The methods of Campens L. and Devereux R. B. were less sensitive, revealing only 5 and 1 patients with aortic root dilatation, respectively. CONCLUSIONS: The results of the study demonstrate that, in order to obtain more reliable information on the condition of the root of the aorta and its proper values, the modified method obtained in the course of the study can be used. This method is more sensitive in detecting the enlargement of the aortic root in CCTD and in diagnosing syndromic CCTD.

THE ANALYSIS OF LGI1 GENE EPIGENETIC ALTERATIONS BY MEANS OF POSTTRANSLATIONAL H3 HISTONE MODIFICATIONS IN MALIGNANT GLIOMAS.

Although there is a progress in understanding the causes and consequences of genetic and epigenetic changes in glioma malignant transformation, many details remain obscure and need further investigation. It is known that process of malignant transformation of gliomas is accompanied by gradual loss of LGI1 gene expression. However, genetic defects causing LGI1 inactivation have not been revealed. In this paper, we have analyzed the LGI1 gene expression in primary cultures of malignant gliomas, and compared these data with epigenetic indicators of transcriptional activity ­ posttranslational H3 histone modifications. We have show the presence of an epigenetic marker of gene repression H3K9me3 near the site of LGI1 transcription initiation in most (5 from 6) studied gliomas. There was not LGI1 expression in these gliomas. Only one glioma showed LGI1 expression, and in this glioma there was no association of LGI1gene with H3K9me3 modification. Thus, we are the first to show a correlation between LGI1 gene expression and the epigenetic indicator H3K9met3 in malignant gliomas. Marker of actively transcribed chromatin Í3K4àñ have not been found in this area of the genome. The data obtained strongly suggest the possibility of gene LGL1 inactivation by epigenetic mechanism: modified «histone code¼.

TOWARD THE OPTIMAL SNP RESEARCH PANEL IN IRRITABLE BOWEL SYNDROME.

The genetic component of multifactorial diseases which include irritable bowel syndrome (1rS) is provided by single nucgetie polmporphism (oNf mliastousual. it is essential to detect the associations of polymorphisms with various patholo- nucleotide are not as us. It cs esed for diagnostic purposes and the development of drug therapies. Analyses of individual polymorphisms have not found their unique relationship with susceptibility to IS. Possibly, several genetic risk factors, together with the influence of the environment cause a synergistic effect leading to the appearance of IgS certain phenotype. This paper presents a research panel of five SNP, which are hereditary factors for violations processes of innate immunity: CD14-159 C> T (rs2569190); TNFa -308 G> A (rs1800629); IL17A -197 G> A (rs2275913); TLR2 Arg753GIn G> A (rs5743708); TLR4 Asp299Gly A> G (rs4986790). Results indicate genetically determined predisposition to l13S as the number of "rare" alleles in the test regions of genes CD14, TNF- and TLR4. Carriage heterozygous genotype GA polymorphism IL17A -197 G>A is also a risk factor for IS in the population studied. On the contrary, carriage of "rare" allele polymorphism Arg753GIn G>A TLR2 gene has to be protective, but normal - predictive properties in the context of the development of lBS.

[Aberrant expression of sox2 gene in malignant gliomas].

Both genetic and epigenetic changes underlite the mechanisms of tumor initiation and progression. In the present study we analyze sox2 gene expression and its epigenetic regulation in primary cultures of malignant gliomas. The sox2 expression was detected in the vast majority (74%) of the investigated gliomas and absent in morphologically normal brain tissue. This indicates the process of glioma malignant transformation. We have also shown that the association of different areas of the sox2 gene with important epigenetic markers, posttranslational modifications of H3 histone H3K4ac and H3K9met3, does not correlate with the sox2 expression. However, this may indicate the stochastic nature of the regulation of sox2 gene expression in malignant gliomas.

[Allogeneic hematopoietic stem cell transplantation for acute myeloblastic leukemia in first remission].

AIM: To evaluate the efficiency of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with acute myeloblastic leukemia in first remission depending on the regimens of conditioning, the source of a graft, and the characteristics of a donor and a recipient. SUBJECTS AND METHODS: In 66 treated patients, including from partially HLA-mismatched relatives (n=4), the efficiency of allo-HSCT from related donors (n=26) and unrelated donors (n=40), were compared. According to cytogenetic findings, 7 (11%), 31 (47%), and 10 (15%) patients belonged to low-, intermediate-, and high-risk groups, respectively. RESULTS: Five-year overall survival (OS) and mortality associated with transplantation were 56 and 22% for allo-HSCT from related donors, 68 and 23% for that from HLA-matched donors, and 71 and 25% for that from partially HLA-mismatched donors, respectively (p=0.8 and p=0.7). The relapse risk after allo-HSCT from unrelated donors was significantly lower than after that from related donors (13 and 35%, respectively; p=0.8). Univariate analysis showed that the OS rates depended on the cytogenetic risk group (OS was 24 and 64% in the high- and intermediate-risk groups, respectively (p=0.027). The relapse risk in chronic graft-versus-host reaction (GVHR) and in grade 3 acute GVHR (p=0.01) was shown to be less than that in grades 1-2 acute GVHR (p=0.06). CONCLUSION: OS rates after allo-HSCT from related and unrelated donors were comparable and unrelated to the source of a graft, the regimen of conditioning, and other characteristics of a donor and a recipient.

[Efficacy of donor lymphocyte infusion in patients after different types of allogeneic hematopoietic stem cell transplantation].

AIM: To evaluate the efficacy of donor lymphocyte infusion (DLI) to prevent and treat recurrences in patients after different types of allogeneic hematopoietic stem cell transplantation (allo-HSCT). SUBJECTS AND METHODS: Data from 118 patients with malignant blood diseases were analyzed. Allo-HSCTs from HLA-matched related donors (n=49), HLA-matched unrelated donors (n=50), partially HLA-matched unrelated donors (n=2), and haploidentical donors (n=24) were performed. The indications for DLI were underlying disease relapse (59 DLIs), resistant disease course (n=40), minimal residual disease (n=1 6), falling donor chimerism (n=1 5), and recurrence prevention (n=1 3). RESULTS: Therapy response was obtained after 57 (44%) DLls. There were 36 (25%) and 30 (21%) cases of acute and chronic graft-versus-host reactions (GVHR), respectively. The use of DLI from HLA-matched donors, its performance in the periods of D+100 to one year after allo-HSCT, a donor chimerism level of over 90% at the moment of DLI, the administration of the initial DLI dose of below 1.10(6) CD3+/kg, and the development of chronic GVHR after DLI were associated with the highest rate of therapy responses. The overall survival rates of patients with DLI were significantly influenced by factors, such as DLI periods, donor chimerism levels at DLI, and the development of chronic CVHR after DLI. CONCLUSION: The choice of the optimal dose of cells, the periods of DLI and its preventive administration improve prognosis in patients after allo-HSCT. The occurrence of acute GVHR is affected by the degree of HLA matching and the type of a donor. The development of chronic GVHR after DLI is associated with the highest rate of responses to DLI and higher survival rates.

[Extracorporeal photopheresis in the treatment of patients with refractory chronic graft-versus-host disease after allogeneic bone marrow transplantation].

AIM: To evaluate the efficiency of extracorporeal photopheresis (ECP) in the treatment of patients with refractory chronic graft-versus-host disease (cGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). SUBJECTS AND METHODS: The study included 49 patients aged 2 to 55 years. Allo-HSCT was carried out in 38 (79%) patients with acute leukemias, 5 (10%) with chronic leukemias, 4 (8%) with myelodysplastic syndrome/myeloproliferative disease, and 2 (3%) with other hematologic diseases. The patients included in the study had glucocorticosteroid (GCS)-refractory disseminated cGVHD or a history of severe complications from GCS therapy. RESULTS: When evaluating the efficiency of therapy, its response was recorded in 37 (77%) cases; the best results were obtained in patients with hepatic (82%), mucosal (76%), and skin (74%) lesions. The mean severity according to the cGVHD Working Group, National Institutes of Health, and a platelet level of more than 100.10(9)/1 were defined as factors improving a therapy response. In the patients receiving ECP, the overall survival was 70%. The latter was higher in the group of patients who had responded to ECP therapy without involving the gastrointestinal tract in the cGVHD process and in those receiving a combination of ECP and other immunosuppressive drugs. CONCLUSION: ECP is an effective treatment for patients with refractory cGVHD, it may be used in those with a history of severe complications from GCS therapy. ECP allows the dose of GCS to be reduced to the point of complete discontinuation.

[Genetic and epigenetic markers of gliomas].

Malignant gliomas, aggressive and highly invasive tumors with a great number of genetic and epigenetic alterations in genes involved in the cell cycle regulation, the apoptotic pathways, cell invasion ability, and angiogenesis, are considered to be among the deadliest of human cancers. The role of epigenetic mechanisms in the pathogenesis of malignant transformation despite recent progress is not yet clear elucidated and remains under intensive study. This review describes the mechanisms of epigenetic regulation of gene expression, including post-translational modification of histones, DNA methylation in the promoter regions, and microRNA regulation. The genetic and epigenetic factors driving the pathogenesis of gliomas in their possible mutual influence and the potential epigenetic targets that can be used in the development of diagnostics and new therapeutic approaches are also discussed.

[Complex chromosome damages in patients with recurrent acute leukemias after allogeneic hematopoietic stem cell transplantations].

AIM: To study the pattern of complex chromosome damages (CCD) in acute leukemias (AL) and their place in the development of post-transplant recurrences (PTR) of AL. MATERIALS AND METHODS: Cytogenetic and partially molecular biological studies of bone marrow cells were conducted in 10 patients with PTR. Of them, 6 patients were diagnosed as having acute lymphoblastic leukemia (ALL), including T-ALL and Ph-positive ALL in 2 and 4 patients, respectively; and 4 patients had acute non-lymphoblastic leukemia (ANLL), including one case secondarily induced by previous polychemotherapy (PCT) and irradiation. The standard G-band staining technique complemented by multicolor fluorescence in situ hybridization in one of the cases was used. RESULTS: It was shown that CCD had the similar pattern in 4 patients before transplantation and in PTR, progressed in 4 more patients, was absent or unnoticed in the early stage of the disease. The other recurrent chromosomal abnormalities that are worthy of notice are as follows: a) the presence of two Ph chromosomes in the cells of two of the 4 patients with Ph+ ALL; b) the frequent involvement of chromosome pairs 9, 19, 5, and 7 into the numerical and structural rearrangements. CONCLUSION: The important feature of PTR of AL is cellular CCDs, a portion of which is clearly related to previous PCT and may be of pathogenetic value for the development of recurrences.

[Study of chromatin decondensation factors in human spermatozoids by flow cytometry].

To date, the mechanisms responsible for radical change of chromatin structure in male germ cells during fertilization are unclear. Evidence suggesting the existence of proteolytic nuclear enzymes in mature human spermatozoids are presented in this work. The possible role of these previously unknown proteases in decondensation of chromatin of spermatozoids in a fertilized ovum is discussed. Application of the flow cytometry technique has shown that treatment of human spermatozoid nuclei with SH-reagents leads not only to destruction of disulfide bonds between protamine molecules that is necessary for their effective utilization but also induces specific endogenous proteolytic activity that consequently results in rather fast decondensation of chromatin followed by proteolytic cleavage of nuclear proteins. A chromatin decondensation process can be almost totally blocked by serine protease inhibitors and components of seminal fluid. An original cytochemical approach of binding of fluorescently labeled protease inhibitor to the target of investigation has been used in order to visualize the localization of proteases in male germ cell nuclei. The results of our study suggest that one of the factors of chromatin reorganization involved in the formation of male pronucleus is endogenous nuclear protease of spermatozoids, which is activated by glutathione or other SH-components of ovum cytoplasm.

[Immunological similarity of diphtheria toxin and EGF receptor].

Cardiomyopathy and neuropathy are the two commonly observed complications in diphtheria patients and in, some instances, individuals vaccinated against diphtheria. The nature of these complications remains not well understood. It was suggested that autoimmunity may play a role in the development of these afflictions. Based on functional similarities between diphtheria toxin (DT) and epidermal growth factor receptor (EGFR), which both can bind to the heparin-binding EGF-like growth factor (HB-EGF) precursors, we suggested that antibodies developed against DT can cross react with EGFR. Here, using serum from healthy donors (n = 10) and diphtheria patients (n = 15), we demonstrated that B-subunit of DT has the antigenic epitopes similar to those of EGFR. Diphtheria toxin as well as EGFR could be recognized by antibodies raised against EGFR and by serum antibodies from diphtheria patients. Moreover serum of diphtheria patients competitively inhibits binding of anti-EGFR antibodies to the receptor. The truncated diphtheria toxin without B-subunit could be detected by serum antibodies of diphtheria patients, but not by anti-EGFR antibodies. Collectively, these studies demonstrate cross-reactivity of antibodies raised against B-subunit of DT and extracellular domain of EGFR and suggest that clinically observed post-diphtheria complications may result from autoimmune inhibition of EGFR function and possible destruction of receptor-positive tissues.

Detection of inflammatory processes during various diseases by the method of flow cytofluorometry.

Oxidative (respiratory) burst is an important manifestation of inflammation. Precise quantitative assessment of this reaction by flow cytometry made it possible to record and evaluate the severity of the inflammatory processes in a wide spectrum of diseases including diphtheria, hepatitis, pneumonia, bronchial asthma, arthritis, vasculitis, postoperative complications, tuberculosis, psoriasis, rheumatoid arthritis, systemic lupus erythematosus, and myocardial infarction. This approach can be employed as a highly sensitive method of detection of inflammatory reactions and monitoring of their course in various pathological processes.

[Analysis of the alpha-synuclein gene dosage variation associated with autosomal dominant form of ParkinsonTs disease].

Fifty-two patients that had ParkinsonTs disease with autosomal dominant type of inheritance were analyzed for the presence of duplications and triplications in exons 4--6 of alpha-synuclein gene using real-time PCR with Taq-Man probes. No mutations involving the examined exons dosage were revealed in alpha-synuclein gene. Thus, mutations modifying copy number of alpha-synuclein gene do not significantly affect the pathogenesis of the autosomal dominant form of ParkinsonTs disease in patients from Russia.

[Possibilities of pharmacological hepatopathy and correction of fat metabolism violations in diabetes mellitus type 1].

The purpose of the research has been the study of possibilities for the pharmacological correction of the morphological changes in liver and lipid metabolism disorders in experimental type 1 diabetes. Simvastatin, new compound from the group of antioxidants--3-hydroxipyridine derivate under the code HP-5, and also the combination of simvastatin and HP-5 have been used for the pharmacological correction in this research. It was revealed that the using of HP-5 and the combination of simvastatin and HP-5 attenuate the order of expression of diabetic hepatopathia as the result of the histological study and hinder from the development of dislipidemia in rates with type 1 diabetes which is more expressed in comparison with the monotherapy by simvastatin. These changes also are followed with the diminishing of the free radical lipid oxidation activity by the specific antioxidant effect of the investigated 3-hydroxipyridine derivate.

[Human RAD51 recombinase: the role in the cell cycle checkpoint and cellular survival].

The RAD51 protein, an eukaryotic homologue of Escherichia coli RecA, plays a central role in both mitotic and meiotic homologous recombination. Here, we demonstrate that short-term silencing of Rad51 gene by specific small interfering (si) RNA induces cell death of the most part of investigated cancer cell lines and normal fibroblasts. Disruption of the Rad51 gene in these cells results in S or (and) G2 cell cycle arrest leading to apoptosis. But some human cancer cell lines demonstrate abolishment of pre-mitotic checkpoint and are not sensitive to siRNA silencing of RAD51 recombinase. Recent experiments show that normal functioning of the recombination repair system is essential for maintenance of genome stability, proliferation of vertebrate cells and, finally, for prevention of dramatic cell death.

[Study of hepatoprotective activity of ethylmethylhydroxypyridine fumarate and lipoate in the experimental toxic hepatitis].

The authors have studied hepatoprotective actions of new derivatives of 3-hydroxipyridine on an experimental model of toxic hepatitis (140 white mice). Mexidol and berlithion are choosed as the preparations of comparison. The method of light microscopy is used for the exploration of morphological changes. The cytolytic contents activity, catalase activity and the level of MDA have been determined in blood serum. The antitoxic effect is valued by the survival of the animals. It is found that all examined bonds is corrected morphological changes in toxic hepatitis and increased the index of animals survival, which is more expressed than the preparations of comparison.

[Analysis of the possible involvement of the glutamate transporter gene EAAT2 and the glutamate receptor genes GRIA1 and GRIA2 in pathogenesis of motor neuron disease in the Russian population].

Polymorphisms of the genes of the glutamatergic system EAAT2, GRIA1, and GRIA2 have been analyzed in patients with sporadic motor neuron disease (MND) from Russia. The disease is not associated with polymorphic alleles of the genes studied, which indicates that EAAT2, GRIA1, and GRIA2 play an insignificant role in the pathogenesis of sporadic MND.

Successful non-myeloablative allogeneic transplantation for treatment of idiopathic hypereosinophilic syndrome.

In this report, we describe two patients with idiopathic hypereosinophilic syndrome (HES) who received a non-myeloablative allogeneic transplantation following a reduced-intensity preparative regimen of melphalan and fludarabine. In both cases, complete donor chimaerism and remission were achieved, and have lasted for more than 10 months. This report provides proof of principle for the feasibility of non-myeloablative transplantation for patients with idiopathic HES, who can show co-morbidity due to eosinophilic infiltration of their organs.