Methods of Synthesis and Antiviral Activity of New 4-Alkyl-3-Nitro-1,4-Dihydroazolo[5,1-c][1,2,4]Triazin-4-ols

[Figure not available: see fulltext.] An azo coupling reaction of α-nitro ketones with 5-diazoazoles was used to obtain 4-alkyl-3-nitro-1,4-dihydroazolo[5,1-с][1,2,4]triazines, which were characterized with respect to their antiviral activity against influenza and Coxsackie B3 viruses. © 2021, Springer Science+Business Media, LLC, part of Springer Nature.This study was funded by the Russian Science Foundation (project No. 20-13-00142)

Проблема химиорезистентности PRAME-экспрессирующей клетки меланомы и способ ее преодоления c помощью бортезомиба

Background. PRAME gene spontaneous expression is frequently observed in a cancer cell. The protein encoded by this gene increases  the viability of tumour cell. NF-κB signalling pathway takes part in PRAME upregulation. It proposes, that stress conditions may increase the expression level of PRAME in the tumour cell and increase cell’s viability after it. We hypothesized that this phenomenon determines chemoresistance of PRAME-expressing cell, which can be overcome by NF-κB inhibitors, such as bortezomib.Materials and methods. We incubated A875 melanoma cells with cisplatin, bortezomib and dexamethasone, as well as with a mixture  of cisplatin with bortezomib and cisplatin with dexamethasone within 24 hours. To assess the cytotoxicity of these combinations MTT-test was used. For evaluation of PRAME expression level, real-time polymerase chain reaction was used. All data were analyzed with Wilcoxon test for coupled samples.Results. It was found that cisplatin and dexamethasone increased an expression level of PRAME compared to control (p <0.03). The addition of dexamethasone to cisplatin reduced cytotoxic effect of cisplatin. Bortezomib has a cytotoxic effect, but it did not increase the activity  of PRAME gene (p = 0.12). PRAME gene activity in cells incubated with a mixture of cisplatin and bortezomib was observed at a lower level in comparison with cells incubated with cisplatin (p = 0.0277).Conclusion. The results of experiments show that an increase of PRAME expression level reduces the sensitivity of melanoma cells to the cytotoxic effect of cisplatin. PRAME activity increases under stress conditions. Using of bortezomib can inhibit the growth of PRAME expression and makes the tumour cell more vulnerable to cytotoxic agents. On the other hand, dexamethasone may increase a resistance  of PRAME-expressing cell to cytotoxic effects.Введение. В настоящее время показано, что активность раково-тестикулярного гена PRAME, характерная только для опухолевой клетки, может контролироваться сигнальным каскадом NF-κB. Белок PRAME увеличивает жизнеспособность опухолевой клетки. Отсюда следует, что стрессовые условия могут повышать уровень экспрессии PRAME и увеличивать жизнеспособность опухолевой клетки. Мы предположили, что данный феномен определяет химиорезистентность PRAME-экспрессирующей клетки. Эту резистентность можно преодолеть ингибиторами NF-κB-пути, такими как бортезомиб.Материалы и методы: инкубирование в течение суток клеток меланомы линии A875 с цисплатином, бортезомибом и дексаметазоном, смесью цисплатина и бортезомиба, а также со смесью цисплатина и дексаметазона. Для оценки цитотоксичности применяемых препаратов использовали МТТ-тест, уровня экспрессии гена PRAME – полимеразную цепную реакцию в реальном времени. Анализ данных проводили с помощью критерия Вилкоксона для связанных выборок.Результаты. Установлено, что цисплатин и дексаметазон увеличивают уровень экспрессии PRAME по сравнению с клетками меланомы линии A875, не подвергнутыми действию экспериментальных веществ (p <0,03). Добавление дексаметазона к цисплатину снижает цитотоксический эффект последнего. Бортезомиб обладает цитотоксическим действием, но практически не увеличивал активность гена PRAME (p = 0,12). В клетках, инкубированных со смесью цисплатина и бортезомиба, активность гена PRAME находилась на более низком уровне по сравнению с клетками, инкубированными с цисплатином (p = 0,0277).Заключение. Результаты экспериментов показывают, что увеличение уровня экспрессии гена PRAME снижает чувствительность клеток к цитотоксическому действию цисплатина. Активность PRAME увеличивается в условиях стресса. Применение бортезомиба препятствует росту уровня экспрессии PRAME и делает опухолевую клетку более уязвимой к цитотоксическим агентам. С другой стороны, дексаметазон может увеличить резистентность PRAME-экспрессирующей клетки к цитотоксическому воздействию цисплатина

Сравнение молекулярно-генетических методов выявления мутаций в гене CALR при миелопролиферативных заболеваниях

Molecular genetic detection of CALR gene somatic mutations is required for myeloproliferative neoplasms diagnosis and treatment according to the novel WHO clinical recommendations. CALR mutations are found in approximately 25–35 % cases of essential thrombocythemia and primary myelofibrosis and they are associated with benign clinical outcome. In this study we have compared sensitivity and selectivity of seve ral different options of CALR mutation molecular genetic detection in blood samples of 379 CMD patients and 17 healthy donors. Among methods compared in our study there have been conventional polymerase chain reaction with electrophoretic detection, real-time quantitative polymerase chain reaction, direct Sanger sequencing of polymerase chain reaction fragments and polymerase chain reaction high resolution melting curve analysis. By means of melting curve analysis CALR mutations have been found in 97 (25.5 %) patients, whereas in the cases of Sanger sequencing and polymerase chain reaction there have been 87 (23.0 %) and 84 (22.1 %) CALR mutation positive patients respectively.Молекулярно-генетические исследования для определения соматических мутаций в гене кальретикулина (CALR) включены в клинические рекомендации Всемирной организации здравоохранения в качестве одних из основных диагностических критериев миелопролиферативных заболеваний. Примерно в 25–35 % случаев эссенциальной тромбоцитемии и первичного миелофиброза бывают выявлены мутации в гене CALR, наличие которых ассоциировано с благоприятным прогнозом течения заболевания. В нашем исследовании выполнено сравнение результатов молекулярно-генетических методов для определения мутаций в гене CALR. Проведен анализ образцов периферической крови 379 пациентов с хроническими миелопролиферативными заболеваниями и 17 образцов крови здоровых доноров. Наличие мутаций в гене CALR определяли методом полимеразной цепной реакции с электрофоретической детекцией и количественной полимеразной цепной реакции в реальном времени, методом секвенирования по Сэнгеру и анализом кривых плавления. Мутации в гене CALR определены у 97 (25,5 %) пациентов методом анализа кривых плавления. Из них у 87 (23,0 %) пациентов мутации в гене найдены методом секвенирования по Сэнгеру. С помощью полимеразной цепной реакции мутации в гене CALR были обнаружены у 84 (22,1 %) пациентов

Clinical and biochemical features of some intravenous iron complexes

Most anemia cases associated with iron deficiency. There are various therapeutic approaches to compensate iron deficiency. In some cases,a rapid restore of body iron is required, which is only possible with intravenous administration. Now a number of intravenous iron preparations are available, and each of them has not only advantages. Considering the drugs side effects, there was a need for drugs with high efficiency, low immunogenicity, and minimal toxicity. One of the decisions was to create preparations based on maltose and isomaltose. Such new intravenous iron preparations are ferric carboxymaltose and iron isomaltoside. Currently, there are no available clinical data that isomaltose and maltose preparations differ significantly with respect to adverse reactions associated with their immunogenicity. Based on study results isomaltose preparations in patients with dextran sensibilization should be used with caution. This is not completely exclude the possibility that both of these drugs can be an immune response trigger with a different specificity than the one on dextran develops. Preparations based on maltose, sucrose and gluconate were neutral in immunoprecipitation assay with dextran-reactive antibodies that determines their preference for patients with dextran sensibilisation. Other important properties of ferric carboxymaltose are: convenience of application and lack of oxidative stress that are determined by the slow iron release

Clinical and biochemical features of some intravenous iron complexes

Most anemia cases associated with iron deficiency. There are various therapeutic approaches to compensate iron deficiency. In some cases,a rapid restore of body iron is required, which is only possible with intravenous administration. Now a number of intravenous iron preparations are available, and each of them has not only advantages. Considering the drugs side effects, there was a need for drugs with high efficiency, low immunogenicity, and minimal toxicity. One of the decisions was to create preparations based on maltose and isomaltose. Such new intravenous iron preparations are ferric carboxymaltose and iron isomaltoside. Currently, there are no available clinical data that isomaltose and maltose preparations differ significantly with respect to adverse reactions associated with their immunogenicity. Based on study results isomaltose preparations in patients with dextran sensibilization should be used with caution. This is not completely exclude the possibility that both of these drugs can be an immune response trigger with a different specificity than the one on dextran develops. Preparations based on maltose, sucrose and gluconate were neutral in immunoprecipitation assay with dextran-reactive antibodies that determines their preference for patients with dextran sensibilisation. Other important properties of ferric carboxymaltose are: convenience of application and lack of oxidative stress that are determined by the slow iron release.</p

Acellbia® and Mabtera® are recognize CD20-positive cells with equal efficiency

In present study were compared characteristics of rituximab produced by Hoffmann–La Roche (Mabtera®) and first domestic biosimilar Acellbia® from Biocad Company. Concentration of protein was measured using Bradford, s method. According to our results, protein concentration in formulations was the same. We electrophoresed formulations in denaturing conditions. Protein from formulations was denatured into fragments. Heavy and light chains of immunoglobulin were observed in gel. Finally, we performed flow cytometry where rituximab was used as primary antibody to detect CD20-positive B-cells of patients with B-cell chronic lymphocytic leukemia. Both Mabtera® and Acellbia® recognized the same number of cells. Thus, assays performed in vitro submitted identity of Mabtera® and Acellbia® characteristics

Microsatellite instability (MSI, EMAST) in the pathogenesis of follicular lymphoma

Background. Genetic instability, an important phenomenon involved in oncogenic transformation and tumor progression, is associated with the insufficiency of the multicomponent DNA repair complex, in particular, the nucleotide mismatch repair (MMR) system. The MMR defect manifests itself as abnormalities in DNA microsatellite repeats, or microsatellite instability (MSI). In the studies of colorectal cancer, the role of MSI in prognostication of the disease, and defining the choice of specific therapy with immune checkpoint inhibitors has been proven.However, in lymphatic system tumors, the significance of this phenomenon is poorly understood. Determination of genetic instability in the onset of follicular lymphoma, a disease characterized by a heterogeneous course, may have prognostic value.Objective: to determine the genetic instability at the onset of follicular lymphoma.Materials and methods. Here we report an analysis of 24 microsatellite repeats and amelogenin loci in tumor cells of 46 follicular lymphoma patients.Results. In the studied cohort, lesions in microsatellite repeats were presented by MSI in 9 cases (19.6 %) and the loss of heterozygosity (LOH) in 19 cases (41.3 %). Most frequent lesions were found for the SE33 marker located at the q14 locus of chromosome 6. A significant association was shown between MSI and the double-hit follicular lymphoma group with rearrangements of the MYC and BCL2/BCL6 genes.Conclusion. Thus, our data indicate that the MSI phenomenon might be involved in the pathogenesis of the lymphatic tumors and particularly follicular lymphoma. However further studies on the expanded cohorts of patients are required to define the possible prognostic value of MSI in lymphatic tumors