Introduction . The incidence of enterovirus meningitis in Ekaterinburg has long exceeded the average in Russia. As a result of sanitary, anti-epidemic and restrictive measures related to the COVID-19 pandemic, there was a decrease in morbidity, which led to a reduction in the circulation of non-polio enteroviruses and created risks after lifting of restrictions. The purpose of the study is to conduct a comparative analysis of non-polio enteroviruses isolated in Ekaterinburg from patients with enterovirus before and after the COVID-19 pandemic. Materials and methods . Genotyping positive samples by sequencing VP1 gene (cerebrospinal fluid, nasopharyngeal, faeces) from patients with aseptic meningitis was performed using the BLAST service. Multiple alignment and phylogenetic analysis were performed using the MEGA software. Results . In 2022, after lifting restrictive measures, the incidence of enteroviruses in Ekaterinburg increased 10 times compared to the previous year (3,390 per 100 thousand inhabitants), but remained 6.5 times lower than the long-term average (22,035 per 100 thousand population). 7 types of non-polio enteroviruses were detected: 2 species A (Coxsackievirus A4, Enterovirus A71) and 5 species B (Coxsackievirus A9 and B2, Echovirus 6, 7, 9). Coxsackievirus A9 dominated, followed by Echovirus 6. Discussion . In 2022, Coxsackievirus A9 prevailed as the predominant type in enterovirus, was not epidemiologically significant and was rarely found in the cerebrospinal fluid samples. Echovirus 6, the second most frequently detected, was characteristic of the Ural Federal District. The remaining viruses (Coxsackievirus A4 and B2, Echovirus 7 and 9, Enterovirus A71) were also detected in patients with enterovirus before the COVID-19 pandemic. Conclusion . After the cancellation of prophylactic measures after the COVID-19 pandemic, the incidence of enteroviruses in Ekaterinburg during 2022 increased due to the restoration of circulation of non-polio enteroviruses.

Introduction. Infant mortality is a universally recognized indicator for social well-being of the population. There is a need for an in-depth analysis of the causes and factors leading to the death of infants at home, including determining the etiology of infectious diseases potentially causing death.The purpose of the study — to determine the implication of herpesviruses in infant out-of-hospital death.Materials and methods. Samples from autopsy material of 23 infants (blood, small intestine, heart, lungs, liver, brain), who died at home from various causes, were analyzed by PCR for the presence of viral DNA and RNA from herpesviruses, orthomyxoviruses, paramyxoviruses, picornaviruses, adenoviruses, coronaviruses, parvoviruses, as well as intestinal viruses.Results and discussion. Positive PCR results were found in 11 cases (43.5 %), of which herpesviruses DNA was detected in 10 cases. In these samples, cytomegalovirus predominated, in association with Epstein — Barr virus and human herpesvirus type 6. This association was detected in blood and internal organs samples from four infants diagnosed with generalized viral infection, and from two infants with acute respiratory viral infection. This suggests the presence of a causal relationship between the lethal outcome and the detection of these viruses. Not all infants showed clinical signs characteristic of generalized herpes infection. In one infant with hypertrophic cardiomyopathy, coronavirus, enterovirus and rotavirus DNA was found in samples of internal organs.Conclusion. Results showed the dominant role of the herpesviruses in out-of-hospital infant death cases.

Relevance. The clinical aspects of tick­borne viral encephalitis (TBE) and Lyme borrelliosis (LB) as monoinfections are well known. At the same time, the issues of interaction of pathogens in tick­borne encephalitis – Lyme borrelliosis mixed infection (TBE – LB mixed infection), the effectiveness of specific prophylaxis, their effect on the incidence rate and clinical forms have not been sufficiently studied.Aims. The aim of this work was to study the clinical course of TBE and TBE ­ LB mixed infection in patients with specific prophylaxis of TBE, as well as in its absence.Materials & Methods. Medical history data from 454 patients patients with a confirmed diagnosis of TBE and TBE – LB mixed infection during 2010–2017 were analyzed.Results. The clinical course of the disease in TBE mono­- and mixed infection, as well as the connection of immunization with the clinical forms of the disease are reflected. The dynamics of the registration of clinical forms of TBE and TBE – LB mixed infection for 8 years was shown using regression models. Synchronization of the incidence of focal and meningeal forms in TBE and TBE – LB mixed infection with a statistically significant decrease in the frequency of their occurrence was noted. The incidence of febrile forms in TBE monoinfection, as well as in mixed infections, remained practically unchanged.Сonclusions. It was found that the incidence of severe focal and meningeal forms of the disease in patients with TBE – LB mixed infection was 1.5 times lower, and the proportion of patients with febrile form TBE was statistically significantly higher than in the group of patients with TBE monoinfection, which may indicate a more favorable during TBE ­ LB mixed infection, and overdiagnosis of febrile form TBE in TBE – LB mixed infection. The increase in the rate of vaccination against TBE of the population contributed to a decrease in the number of TBE и TBE – LB mixed infection cases due to severe clinical forms (focal and meningeal).

Treatment ofprognostically unfavorable forms ofacute leukemia isaserious problem ofhematology, since their frequency inthe group ofadult patients reaches 60%, and inelderly patients- 80-90%.Aim: toanalyze the case ofdiagnostics and treatment ofacute myelomonoblastic leukemia with anunfavorable genetic prognosis inayoung adult female using double haploidentical bone marrow transplantation.Samples ofbone marrow and peripheral blood ofpatient K., 35years old, who recived program chemotherapy and double haploidentical bone marrow transplantation inthe Sverdlovsk Regional Hematological Centre were examined.Atthe onset ofdisease was noted hyperleukocytosis, blastaemia and infiltration ofleukemic cells inthe bone marrow. Cytochemical reactions tolipids were positive in7,0% ofblasts, small-granular and large-granular glycogen was detected in24,0% ofblast cells, and diffuse- in22,0%. Immunophenotypically, 28,0% ofblast cells were characterized byexpression ofHLA-DR, CD13, CD33, CD34, CD38, CD117, MPO-cyt, 13,0%- CD33, CD11cyt, CD64, CD14. The cytogenetic study determined the karyotype 48, XX, +4, +21[2]/ 47, XX, +4[3]/ 46, XX[2]. Non-synonymous transversions с. 2447A>Тinс-KIT gene, с.215С>GinТР53, and transition с. 2644С>ТinDNMT3A were detected using direct sequencing method. Despite the detection ofoncogenic mutations, the tumor was chemosensitive topolychemotherapy (including anthracyclines with cytarabine). However, given the unfavorable genetic prognosis, itwas decided toperform abone marrow transplantation. Due tothe lack ofanHLA-matched unrelated donor, the first haploidentical transplantation was performed from the patient'seldest daughter. At+51day, transfusion ofdonor white blood cells with correction ofthe immunosuppression scheme was performed tocorrect persistent pancytopenia. At+ 71day after the first transplantation, taking into account the remaining pancytopenia, asecond haploidentic transplantation from the patient'syounger daughter was performed. The total duration ofhematological remission was 48months, including 36months after the second haploidentical bone marrow transplantation.

This work evaluates the results of long-term dynamic monitoring of non-polio enteroviruses (NPEV) circulation in the indicator subpopulation in Yekaterinburg (children aged 3 to 6 years old), isolation of this group of viruses from specimens of sewage and the spectrum of causative agents detected in the liquor of patients with enteroviral meningitis (EVM). We established a high comparability rate of Coxsackie B and ECHO virus serotypes etiologically significant in EVM and isolated from healthy carriers. NPEV detected in sewage waters over the observation period were presented mainly by Coxsackie B viruses (84.0 %), while ECHO viruses (75.6 %) dominated among the causative agents of EVM. A conclusion was made about low informative value of the sewage study results for evaluation and forecast of the EVM epidemic situation as well as about the rationale for including the screening studies of NPEV circulation in the indicator subpopulation into the system of virological and molecular genetic monitoring.

Aim: to analyze the case of immunological detection and treatment of young male with resistant acute myeloid leukemia in Sverdlovsk Regional Hematological Centre. Bone marrow and peripheral blood samples obtained from male, aged 38, treated using intensive chemotherapy programs and then obtained supportive care and palliative chemotherapy in municipal hospital. Immunohistochemical characterization of AML blast cells: medium and small cells with one or several nucleoli, MPO, CD34, CD117, CD68 positive. In cytochemistry lipids were positive in 29.0% of blasts, glycogen – 81.0%. Immunophenotype of blast cells was CD45, CD13, CD33, CD34, CD38, CD117, MPO-cyt (+). Karyotype was 47, XY, +13 and changed during AML clonal progression. In case, two non-synonymous substitution were co-existed in samples: NRAS gene с. 182 A>C and TP53 gene с. 215 С>G. The number of NRAS gene point mutations changed during AML clonal progression (additional c. 35 G>С substitution was found). Immunophenotype of blast cells not changed during progression. Overall time of patient observation was 42 week.

Sverdlovsk region is a zone with a strenuous natural-anthropogenic focus of virus tick-borne encephalitis (TBE). Incidence with the disease has decreased by 5 times over the last 20 years due to mass vaccination among population. Since 2015 incidence with TBE has remained steady at fewer than 3 cases per 100,000 people. However, over the last 10 years incidence with TBE has been decreasing at a significantly slower rate due to a considerable growth in number of immune people (from 68 % in 2007 to 84.99 % in 2018). Analysis revealed that probability of the disease after a person had been bitten by a tick was quite different on different territories in the region. Our research goal was to develop a procedure for ranking administrative territories as per risks of clinical TBE occurrence among people bitten by ticks. We took a number of people bitten by ticks per one TBE case as our risk parameter. Our analysis revealed that average regional risk reached its maximum values (1:40–1:50) in years prior to implementation of mass vaccination against TBE. As a number of immune people grew, risk fell by 6 times (just 1 TBE case per 319 bitten people in 2018). Average regional risk was taken as to be equal to 1. We ranked administrative territories as per their risk index values (a ratio of a territorial risk to average regional one). We showed that ranking of TBE-endemic territories as per their risk index allowed implementing a differentiated approach to planning and organizing efficient prevention.

The aim of the study was to assess pathogenetic significance of TP53 gene mutations in adult acute myeloid leukemia (AML) patients. Clinical observation was carried out on 114 AML patients at the Sverdlovsk Regional Clinical Hospital No. 1 (Ekaterinburg), including 56 males and 58 females. The average age of subjects was 53.3±2.8 years. Morphologically, AML was previously verified in all cases at specialized laboratories by using standard cytological, cytochemical, immunophenotypic, histological and immunohistochemical methods. The study included the following variants of AML: M0 – 5, M1 – 9, M2 – 47, M2baso – 3, M2eo – 2, M3 – 8, M4 – 25, M4eo – 3, M5 – 3, M6 – 4, M7 – 1, acute myelofibrosis – 1, blastic plasmacytoid dendritic cell neoplasm – 2. Samples of peripheral blood and bone marrow aspirates from patients were examined. Exons 4-11 within the TP53 gene were tested for molecular damage by using sequencing method. In addition, 81 samples, including 22 AML with normal and 23 with an unspecified karyotype were examined for gene mutations by using molecular genetic and immunohistochemical methods. cDNA sequencing was carried out on automatic genetic analyzer in forward and reverse sequences. The sequencing results were processed by using the MEGA X software and statistical hypothesis that they may be described by a binomial distribution. The statistical hypothesis was tested by using Fisher’s exact test and χ2 test. According to the results of cytogenetic and PCR studies, a favorable prognosis was determined in 25 cases (21.9%), intermediate – 24 (21.1%) and unfavorable – in 33 (28.9%). No genetic abnormalities could be detected in 32 samples (28.1%) with standard cytogenetics and real-time PCR, and prognosis option for such patients was not specified. TP53 missense mutations were revealed as C292T, A377G, A659G, C817T transitions (4 cases) and C569G, G733T, G841C transversions (3 cases); synonymous A639G substitutions were also determined (1.8% ) and C891T (0.9%), in codon position 3, providing no pathogenetic significance. In one sample (0.9%), a deletion of thymidine at position 645 of the coding sequence was determined, leading to produced shortened mutant protein. All the above mutations were localized in the region of the DNA-binding domain. Also, in one case (0.9%), a tandem duplication of 19 nucleotides at position 960 of the coding sequence of the NLS domain protein located in acetylation site. Non-synonymous C215G transversion, which is a polymorphic gene variant, was determined in 94 samples (82.5%). Clinically, all TP53-positive AML were characterized by unfavorable prognosis and primary resistance to standard chemotherapy. The average age of such patients was 63.0±5.4 years, with average follow-up reaching up to 3.1±0.9 months.

Extrapolation prediction of epidemic situation as per tick-borne viral encephalitis (TVE) on endemic territories that is based on analyzing time rows of morbidity is a promising approach to be applied in predictive medical-ecological and epidemiologic research. The authors examined long-term dynamics showing both number of people who suffered from tick bites and morbidity with tick-borne viral encephalitis (TVE) in 4 regions in the Ural Federal District over 2007–2017. We applied a sum of harmonic functions as a mathematic model; parameters of the functions were detected with Le-venberg–Marquardt procedure for non-linear estimates. The technique is flexible and it allows both to apply parameters of harmonic fluctuation that are common for all 4 regions and to estimate parameters that differ in various regions and are of special interest (average long-term values and other fluctuation parameters). One of the research goals was to estimate dynamics in number of people who suffered from tick bites and morbidity with TVE in the Ural Federal District regions over the examined period and to predict epidemiologic situation for the coming years. To do that, we built several harmonic regression models with different number of estimated parameters. To compare and rank the models, we applied Akaike consistent information criterion that determines optimality as a compromise between a model accuracy and complexity. Our analysis of morbidity with TVE over 2007–2017 in Sverdlovsk, Chelyabinsk, Tyumen, and Kurgan region allowed us to quantify discrepancies in average long-term parameters between these Ural Federal District regions. The highest average long-term morbidity was fixed in Kurgan region; the lowest one, in Sverdlovsk and Chelyabinsk region. But a number of people who suffered from tick bites was higher in Sverdlovsk, Chelyabinsk, and Tyumen region than in Kurgan region over the same period. We showed that long-term fluctuations in ticks activity in the Ural Federal District can be considered in-phase and it can possibly mean there is regional synchronization. We detected quasi-periods of cycles both for number of people bitten by ticks and morbidity with TVE and built a short-term prediction for epidemic situation as per TVE in the region on the basis of the proposed harmonic model for a period up to 2022; a probable TVE morbidity peak can be reached in 2020–2021.

The aim of this study was to determine the role of the human neonatal receptor for the Fc fragment of IgG (hFcRn) as a common uncoating cellular receptor for echoviruses and coxsackievirus A9 during infection of human rhabdomyosarcoma (RD) cells. Material and methods. The protective effect of the human serum albumin, purified from globulins, (HSA-GF) and antibodies to hFcRn was studied in RD cells infected with several strains and clones of species B enteroviruses possessing different receptor specificity (echoviruses 3, 9, 11, 30 and coxsackieviruses A9, B4, B5). Results. It was shown that HSA-GF at concentrations of 4% or less protected RD cells from infection with echoviruses 3, 9, 11 and coxsackievirus A9. The antibodies to hFcRn at concentrations of 2.5 ug/mL or less demonstrated the similar spectrum of protective activity in RD cells against infection with echoviruses 3, 9, 11, 30 and coxsackievirus A9. The protective effect of HSA-GF or the antibodies to hFcRn was not observed in RD cells infected with coxsackieviruses B4 and B5 that need coxsackievirus-adenovirus receptor for uncoating. Discussion. The usage of the previously characterized echovirus 11 clonal variants with different receptor specificity allowed us to define the function of hFcRn as a canyon-binding uncoating receptor in RD cells. The kinetics and magnitude of the observed protective effects correlated with receptor specificity of the enteroviruses used in this work supporting the two-step interaction of DAF-dependent echoviruses with the cellular receptors. Conclusions. In this study, the function of hFcRn was defined in RD cells as a canyon-binding and uncoating receptor for echoviruses and coxsackievirus A9. The two-step interaction of DAF-dependent echoviruses during entry into the cells was confirmed: initially with the binding receptor DAF and subsequently with the uncoating receptor hFcRn.

Tick-borne encephalitis (TBE) as an infectious disease remains a significant issue in the Urals Federal District (the UFD). To correctly describe impacts exerted by risk factors on TBE epidemic process, it is necessary to analyze both common and individual peculiarities related to how TBE epidemic process develops on endemic administrative territories. We assessed impacts exerted by biological and social factors on morbidity with TBE in four endemic regions in the UFD (Sverdlovsk, Chelyabinsk, Tyumen, and Kurgan regions) over 2007–2017. To quantitatively assess contributions made by specific factors into morbidity with TBE, we calculated chances for people who suffered tick bites to fall ill with TBE; it allowed us to apply standard procedures within generalized linear models theory (GLM), namely logistic regression. Our analysis included aggregated data on quantity of people who were bitten by ticks and fell will with TBE in all the examined regions. We also assessed data for each specific region as all these endemic territories had both common and specific regularities related to TBE endemic process development. We showed statistically significant impacts exerted by specific manageable risk factors (vaccination, immune prevention, and acaricide treatment performed on a territory) on a possibility of TBE occurrence among people bitten by ticks on endemic territories. The examined UFD regions differ as per effects produced by natural and social risk factors on TBE development. Mass vaccine prevention is a key factor in the control over morbidity with the disease. Risk-oriented approach provides significant additional data for analyzing an epidemiological situation and planning efficient preventive activities in TBE natural foci.

V.A. Mishchenko1,2, O.V. Ladygin1, I.P. Bykov1, J.A. Zakharova1, A.G. Sergeev1,3, I.A. Kshnyasev2 Rospotrebnadzor's Yekaterinburg Research Institute of Viral Infections, 23 Letnyaya Str., Yekaterinburg, 620030, Russian Federation Institute of Plant and Animal Ecology of the Urals Department of Russian Academy of Science, 202 8 Marta Str., URAN, Yekaterinburg, 620144, Russian Federation Ural State Medical University of the RF Public healthcare Ministry, 3 Repina Str., 630028, Yekaterinburg, Russian Federation

The molecular genetic landscape of acute myeloid leukemia (AML) have specific features in elderly patients, and these features correlates with hematopoietic progenitor cells senescence. Aim: to estimate the frequency of mutations in DNMT3A, FLT3, KIT, NPM1, NRAS, TP53 and WT1 genes in AML patients in elderly. Bone marrow and peripheral blood samples obtained from 54 AML patients aged over 60 years old. Distribution of the patients according to FAB-classification was as follows: AML M0 - 2, M1 - 6, M2 - 27, М3 - 2, M4 - 11, M5 - 1, M6 - 3, acute myelofibrosis - 1, blastic plasmacytoid dendritic cell neoplasm - 1. Detection of mutations in DNMT3A, FLT3, KIT, NPM1, NRAS, TP53 and WT1 genes performed by automatic direct sequencing technique. In the study group were more common patients with unfavorable (33.3%, n=18) and unspecified (42.6%, n=23) cytogenetics. The average frequency of functionally significant mutations in all investigated genes among the treated AML patients was 38.9% (n=21), including 3 cases (27.3%) with normal karyotype, 11 cases (61.1%) with unfavorable cytogenetics, 7 cases (30.4%) with unspecified karyotype. Average frequency of mutations in TP53 gene exons 4-11 was 20.0%, FLT3 gene exons 12-15 and 19-21 18.4%, DNMT3A exons 18-26 - 9.1%, NRAS gene exons 1-4 - 7.7%, KIT gene exons 7-12 and 16-19 - 5.9%, NPM1 gene exons 9-12 - 5.4% (n=2), DNMT3A - 9.1% (n=1). Multiple point mutations in investigated genes were detected in 11.1% AML specimens (n=6, usually FLT3 gene mutations, including FLT3 ITD in 4 cases). Additional gene mutations detection using direct sequencing allowed to clarify the prognostic stratification of AML from groups of unspecified and intermediate prognosis in 35.9% (n=10). In all cases, they were associated with an unfavorable prognosis. Thus, using of cytogenetic and additional molecular genetic research, a favorable prognosis of overall survival was established in 2 cases (3.7%), intermediate - in 9 cases (16.7%), unfavorable - in 27 cases (50.0%), and unspecified - in 16 (29.6%).

The frequency of acute myeloid leukemia (AML) increases with age, respectively, the range of identified gene mutations and the pathways involved carcinogenesis can vary and affect the prognosis of treatment. The aim of the study was to estimate the frequency of mutations in DNMT3A, FLT3, KIT, NPM1, NRAS, TP53 and WT1 genes in acute myeloid leukemia (AML) patients (pts) aged 15-45 years old using direct automatic sequencing technique. Bone marrow and peripheral blood samples obtained from 36 AML pts aged 15 to 45. Distribution of the pts according to FAB-classification was as follows: AML M0 - 2, M1 - 1, M2 - 15, М3 - 2, M4 - 11, M4eo - 2, M5 - 2, blastic plasmacytoid dendritic cell neoplasm-1. Detection of mutations in ASXL1, DNMT3A, FLT3, KIT, NPM1, NRAS, TP53 and WT1 genes performed by automatic direct sequencing technique. The average frequency of functionally significant mutations in all investigated genes among the treated AML pts was 41,7 % (n=15), including 6 cases (40,0 %) with unfavorable cytogenetics, 6 cases (54,5 %) with normal karyotype, 3 cases (37,5 %) with favorable cytogenetics. These data correspond to the average frequency of point mutation in AML with normal and abnormal karyotype. Average frequency of mutations in FLT3 gene exons 12-15 and 19-21 - 21,9 %, NRAS gene exons 1-4 - 13,0 %, WT1 gene exons 6-9 - 11,1%, NPM1 gene exons 9-12 was 10,7 %, KIT gene exons 7-12 and 16-19 - 10,0 %, DNMT3A exons 18-26 - 7,1 %, TP53 gene exons 4-11 - 0,0 %. Multiple point mutations in investigated genes detected in 13.9 % AML specimens (usually KIT gene non-synonymous substitution c. 1621 А>С). Cryptic gene mutations detection using direct sequencing technique allowed to clarify the prognostic stratification of AML from groups of favorable and intermediate prognosis in 36,8 % (n=7). Thus, using of cytogenetic and additional molecular genetic research, a favorable prognosis of overall survival was established in 6 cases (16,7 %), intermediate - in 10 cases (27,8 %), adverse - in 18 cases (50,0 %), and unspecified - in 2 (5,6 %).

Экспериментально установлено влияние роданина на форму графика инфекционной активности в одиночном цикле репродукции эховируса 11 серотипа в культуре клеток RD. Выявлено более выраженное ингибирующее действие роданина на процесс депротеинизации гемагглютинирующего клона эховируса 11 по сравнению с негемагглютинирующим клоном. Представлена динамическая модель ин- терпретации графиков одиночного цикла репродукции эховирусов, позволяющая проводить сравни- тельный анализ задержки интернализации, интенсивности депротеинизации и эффективности синтеза новых вирионов.