Plant pathogens present a major challenge to crop production, leading to decreased yield and quality during growth and storage. During long-term storage, healthy onions can develop diseases from latent pathogen infections. This poses a challenge for onion growers because infected bulbs without visible symptoms can lead to significant crop losses during the growing season. In this study, we aimed to isolate and identify Fusarium species from yellow onion bulbs (Allium cepa L.) that developed disease symptoms during storage. The aggressiveness of these strains against onion bulbs and seedlings was also evaluated. The isolated strains were further subjected to morphological and molecular differentiation. The results revealed that all 16 isolated strains belonged to the Fusarium complex species incarnatum-equiseti and Fusarium fujikuroi, namely, F. proliferatum (98%), F. oxysporum (1%), and Fusarium sp. (1%). Koch’s postulate analysis of isolated strains revealed varying aggressiveness on onion bulbs and plants depending on fungal species. Disease symptoms developed more slowly on plants than on onion bulb plants according to Koch’s postulates. Moreover, the results revealed that Fusarium strains that can infect onion plants were less pathogenic to onion bulbs and vice versa. In addition, three isolates were found to be non-pathogenic to onions. Furthermore, the in vitro control of Fusarium species through Bacillus velezensis KS04-AU and Streptomyces albidoflavus MGMM6 showed high potential for controlling the growth of these pathogenic fungi. These results may contribute to the development of environmentally friendly approaches for controlling onion spoilage caused by pathogens during storage.

The first Russian settlers developed relatively large cities and a new phenomenon called 'urban habitat' to the north of Western Siberia. This article presents the results of a study on soil samples taken from the cultural layer of Berezov city during excavations of the city's kremlin, or civic centre, posad, or town settlement area, and the farmstead within the posad. The samples are dated from the XVI to the end of the XVIII century. Examination of the samples revealed 12 types of parasite eggs from both humans and animals. The city's cultural layer was found to be relatively highly contaminated with parasite eggs. The species composition of parasite eggs in samples collected from the cultural layer varied according to land use, i.e. whether public, communal, or livestock, or sanitary condition in Berezov city. An analysis of historical and archaeological data enabled the reconstruction of certain nutritional aspects of the population in Berezov city at the time. Additionally, certain cultural and abiotic factors contributing to the high parasite infestation of humans and domestic animals were identified, and similarities can be found in the research of other Russian cities of Siberia in the XVI-XIX centuries.

Systemic lupus erythematosus (SLE) is characterized by a systemic dysfunction of both the innate and adaptive immune systems, leading to an attack on healthy tissues of the body. During the development of SLE, pathogenic features, such as the formation of autoantibodies against self-nuclear antigens, cause tissue damage including necrosis and fibrosis, with increased expression levels of the type Ⅰ interferon-regulated genes. Standard treatments for lupus with immunosuppressants and glucocorticoids are not effective enough but cause side effects. As an alternative, more effective immunotherapies have been developed, including monoclonal and bispecific antibodies that target B cells, T cells, co-stimulatory molecules, cytokines or their receptors, and signaling molecules. Encouraging results have been observed in clinical trials with some of these therapies. Furthermore, a chimeric antigen receptor T cell therapy has emerged as the most effective, safe, and promising treatment option for SLE, as demonstrated by successful pilot studies. Additionally, some emerging evidence suggests that gut microbiota dysbiosis may significantly contribute to the severity of SLE, and the normalization of the gut microbiota through methods such as fecal microbiota transplantation presents new opportunities for effective treatment of SLE.

The efficacy of chimeric antigen receptor (CAR)-T therapy may not match initial expectations due to the influence of multiple circumstances, some of which cannot be predicted. CAR-T treatment groups include high-risk patients, particularly those with TP53 mutations. A significant body of research has demonstrated that mutations in the TP53 gene play a pivotal role in cancer development and progression. Any aberration in the TP53 gene in cancer is invariably associated with complications and a poor prognosis. Moreover, mutations in the TP53 gene have been observed to correlate with resistance to conventional chemotherapy, prompting the use of alternative therapeutic approaches, including CAR-T therapy. However, there is a possibility that abnormalities in the TP53 gene may affect patients after CAR-T cell administration reducing the efficacy of therapy. This review examines the link between TP53 mutations in cancer and the efficacy of CAR-T therapy, as well as the potential implications of this aspect in therapeutic planning.

Болезнь Вильсона-Коновалова - редкое аутосомно-рецессивное заболевание, которое характеризуется патологическим накоплением меди в печени, головном мозге и других тканях. Дифференциальная диагностика болезни Вильсона-Коновалова представляет собой сложную задачу вследствие выраженной гетерогенности клинических проявлений. Это подчеркивает важность разработки как новых методов диагностики, так и усовершенствования существующих. В рамках настоящего исследования было проведено сравнение клинической диагностики заболевания с результатами молекулярно-генетических исследований. Проанализировано 42 пациента с подозрением на болезнь Вильсона-Коновалова. Произведена оценка значения биохимических показателей метаболизма меди (концентрация церулоплазмина, щелочной фосфатазы, общего билирубина, АСТ, АЛТ сыворотки крови, содержание меди в печени, экскреция меди с мочой) согласно Лейпцигской количественной шкале. Для молекулярно-генетического анализа использовали геномную ДНК. Обогащение интересуемых регионов генома проводилось с помощью ПЦР длинных фрагментов. Для подготовки ДНК библиотек был использован набор Nextera DNA Flex (Illumina, США). Секвенирование проводилось на приборе Illumina MiSeq (Illumina, США). В результате исследования в 62,5% случаев у пациентов, направленных на подтверждение диагноза (по Лейпцигской количественной шкале), были найдены мутации в гене ATP7B, что подтверждает ценность комплексной диагностики по Лейпцигской количественной шкале с учетом клинической симптоматики и лабораторных показателей метаболизма меди. Wilson’s disease is a rare autosomal recessive disorder characterized by abnormal accumulation of copper in the liver, brain, and other tissues. Wilson’s disease differential diagnosis is a difficult task due to the pronounced clinical heterogeneity. This emphasizes the importance of developing both new diagnostic methods and improving existing ones. As part of this study, we compared clinical diagnostics with the results of molecular genetic studies. We analyzed 42 patients with suspected Wilson’s disease. The biochemical parameters copper metabolism values were assessed (serum ceruloplasmin concentration, liver copper content, urinary copper excretion, alkaline phosphatase, total bilirubin, AST, ALT) according to the Leipzig quantitative scale. We used genomic DNA for molecular genetic analysis. Regions of interest in the genome was enriched using long-range PCR. The Nextera DNA Flex kit (Illumina, USA) was used to prepare DNA libraries. Sequencing was performed on an Illumina MiSeq device (Illumina, USA). As a result of the study, in 62.5% of cases in patients aimed at confirming the diagnosis (according to the Leipzig quantitative scale), we found mutations in the ATP7B gene, which confirms the value of a comprehensive diagnosis according to the Leipzig quantitative scale, taking into account the clinical symptoms and copper metabolism laboratory parameters.

The nuclease from S.marcescens heads a family of homological nonspecific nucleases. Its cultivation features, structure, mechanism of action, some physical, chemical and biochemical properties are well studied. It is presented by two isoforms, differing by N-terminal fragment and some properties. The nuclease demonstrated a cancer suppressing effect in mice. It is a key component of the anti-rabies composition which increases survival of the infected mice. It is similar by its efficiency to Pulmozyme® that is used for airway cleansing during the treatment of pulmonary diseases. The aim of the study was to analyze a cytotoxic effect of S. marcescens nuclease and its separate isoforms at different concentrations. To determine the cytotoxicity we used MTT assay and human cell cultures of colorectal cancer -HCT 116, breast cancer MCF-7 and normal skin fibroblasts. Results show that the nuclease demonstrates a weak cytotoxic effect at the amounts of 0.2 – 25 µg per 1 ml of the cell culture medium that is close to its therapeutic doses. This effect is not connected with the loss of the enzymatic activity. The isoforms are similar by their IC50.

Using massively parallel sequencing, the COL1A1 and COL1A2 genes were analyzed in 16 patients from 10 families with osteogenesis imperfecta types I, III, and IV. To analyze the mutations in these genes, a panel of primers was developed for sequencing the complete gene sequence. As a result of the work, 10 mutations were revealed: six of them are in the COL1A1 gene, four mutations in the COL1A2 gene. All mutations, except one, were previously described in the literature and were found in patients with various types of osteogenesis imperfecta. Missense mutations were identified in five families, nonsense mutations in two families, splice site mutations in two cases, and a frameshift mutation in one patient. Unique mutations that were not repeated in unrelated patients were found in all families. The revealed high heterogeneity of the spectrum of mutations in the COL1A1 and COL1A2 genes in osteogenesis imperfecta indicates the effectiveness of using MPS-based methods for the diagnosis of this pathology.

Болезнь Вильсона-Коновалова (БВК) - аутосомно-рецессивное заболевание, развивающееся вследствие накопления меди в организме при повреждениях гена АТР7В. В настоящем исследовании проводился поиск мутаций в этом гене методом массового параллельного секвенирования у больных с БВК. Для целевого обогащения интересуемых регионов была разработана панель праймеров для ПЦР длинных фрагментов. У 6 пациентов из 12 проанализированных выявлены патогенные и вероятно патогенные варианты нуклеотидной последовательности гена АТР7В. Полученные результаты указывают на то, что разработанный метод таргетного массового параллельного секвенирования позволяет эффективно выявлять мутации в гене ATP7B. Wilson’s disease is an autosomal recessive disease that develops as a result of the accumulation of copper in the organism when the ATP7B gene is damaged. The present study searched for mutations in this gene using massively parallel sequencing in patients with Wilson’s disease. For targeted enrichment of the regions of interest, a primer panel for PCR of long fragments was developed. In 6 patients out of 12 analyzed, pathogenic and probably pathogenic variants of the nucleotide sequence of the ATP7B gene were identified. The obtained results indicate that the developed method of targeted massively parallel sequencing allows efficient detection of mutations in the ATP7B gene.

Детская клиническая больница № 1, ярославль 150003, Российская Федерация 36 Клиническая больница № 2, ярославль 150010, Российская Федерация Цель исследования.Изучить особенности и разнообразие спектра патогенных генетических вариантов гена CFTR (ABCC7) у российских пациентов с МВ, представленных в Регистре больных муковисцидозом (МВ) Российской Федерации (РФ) 2017г.Материал и методы.Проанализированы результаты генотипирования, включавшего анализ частых патогенных генетических вариантов, секвенирование кодирующей последовательности, поиск генных перестроек гена CFTR, 3096 больных из 81 региона-субъекта Российской Федерации, представленных в Регистре больных МВ в РФ 2017 г.Результаты.Выявлено 196 патогенных генетических вариантов гена CFTR.Суммарная доля 11 генетических вариантов c.1521_1523delCTT (F508del), c.54-5940_273+10250del21kb (CFTRdele2,3), c.274G>A (E92K), c.2012delT (2143delT), c.3718-2477C>T (3849+10kbC->T), c.3846G>A (W1282X), c.2052_2053insA (2184insA), c.1545_1546delTA (1677delTA), c.3909C>G (N1303K), c.1624G>T (G542X), c.413_415dupTAC (L138ins) составляет 75,6 %. 102 редких вариантов обнаружены однократно, 29 -дважды.Как в спектре, так и по частоте у пациентов в РФ преобладают варианты, приводящие к серьезным нарушениям функции белка CFTR (I, II, III классы).44 генетических варианта не внесены в базы CFTR1 и CFTR2.Заключение.На основании данных Регистра 2017 года определены спектр и относительные частоты патогенных вариантов последовательности гена CFTR у российских больных МВ; описано их распределение в зависимости от класса и типа.Выявлены генетические варианты, ранее не описанные в базах CFTR1 и CFTR2.Полученные результаты могут использоваться для оптимизации генетического консультирования и клинической работы с семьями, отягощенными МВ, а также для дальнейших исследований патогенетической значимости ранее не описанных генетических вариантов гена CFTR.

The studies have been carried out on the Afrikanda (the Murmansk region) titanium-containing minerals segregation by magnetic separation. The technological scheme of ore preparation and further enrichment is developed taking into account the dissemination and disclosure of a specific mineral for obtaining both titanium-magnetite and perovskite concentrate. Mineralogical characteristics of the crushed ore of the deposit have shown a high degree of titanium-magnetite disclosure. The first stage of magnetic separation of crushed ore is an effective method for isolating titanium-magnetite. The magnetic separation of crushed ore excludes the re-grinding of titanium-magnetite, since fine-grained perovskite perforation provides for finer further grinding, and also reduces the amount of ore mass entering the grinding process in the perovskite release cycle. The nonmagnetic fraction is represented by olivine, perovskite and also slightly calcite. The analysis of the change in the granulometric composition of the ore as a function of the grinding time has been performed. At various fineness of grinding by magnetic separation, conditioned titanium-magnetite concentrates have been obtained. The yield of the concentrate from the ore is 10.9–10.5 %, with Feобщ content equal to 60.6–62.0 %. Perovskite has a weakly expressed magnetic susceptibility with respect to titanium-magnetite. Studies have been carried out on the preparation of perovskite concentrate by magnetic separation. The influence of the degree of grinding of the separation feed, desliming and preliminary classification on the separation of perovskite by magnetic separation has been shown. Perovskite concentrates with the content of 44–46 % TiO2 have been obtained. The indexes of enrichment of titanium-magnetite-perovskite ore by magnetic separation have been presented to obtain titanium-magnetite and perovskite concentrate. An important role in obtaining quality concentrates is associated with the preliminary preparation of ore – grinding, desliming and classifying

One of the approaches to effective airway cleansing is the degradation of DNA into smaller fragments. For this purpose Pulmozyme® is used with high efficacy because it contains recombinant DNase I as its active component. The aim of the study was to comparatively analyze DNase activity of Pulmozyme® and the nuclease from gram-negative bacteria Serratia marcescens, because at optimal conditions the catalytic efficiency of the nuclease is much higher than the efficiency of DNase I. Highly polymerized DNA and purulent-mucous sputum were used as substrates. The examination showed that both S. marcescens nuclease and Pulmozyme® hydrolyzed DNA in sputum. Also S. marcescens nuclease was found capable of hydrolyzing DNA in conditions that are standard for Pulmozyme® and suitable for its therapeutic application. For manifesting the similar hydrolytic activity the nuclease amount in the assay mixture containing highly polymerized DNA or the sonicated sputum and NaCl together with calcium- or magnesium- cations can be about 10- time lower than that of the recombinant DNase I. In the presence of magnesium cations the DNase activity of both S. marcescens nuclease and Pulmozyme® was higher than in the presence of calcium cations.

The main goal of the investigation was to analyze the mechanism of Mg2+ action on the digestive activity of Serratia marcescens nuclease towards RNA due to the potential application of the nuclease as a reagent for removing nucleic acids from biomedical samples as well as an antiviral factor. Examination of the mechanism revealed that it was similar to the mechanism of the metals actions on the DNase activity. The optimal Mg2+ amount was linked with the changing secondary structure of RNA—substrates within A-helix. Addition of Mg2+ was found to affect both the rates of products dissociations from the enzyme-substrate complexes and the enzyme associations with the substrates, that was supported by strong increase in the Kcat values and change in the Km values. Comparing the mechanisms of Mg2+ action on RNase and DNase activities in S. marcescens nuclease, we identified a preference for DNA in the absence of Mg2+ that was supported with the Kcat values.

The article presents data on separation of minerals of eudialyte ore with low magnetic susceptibility using a high-intensity wet magnetic separator and pulp pulsation. The optimal separator variables of the magnetic field inductance, pulp pulsation and matrix filler diameter to ensure maximum efficiency of processing are determined.

Cowden syndrome is a rare genetic disorder caused by the abnormal development of all three germ layers. Taking into consideration the mesodermal origin of mesenchymal tissue that gives rise to the vascular system, the pathology of cerebral blood vessels can occur in conjunction with this disease and cause the development of cerebrovascular disease in such patients.