МОРФОФУНКЦИОНАЛЬНЫЕ ОСОБЕННОСТИ АОРТАЛЬНОГО ГРАФТА ПОСЛЕ ДЕЦЕЛЛЮЛЯРИЗАЦИИ

Purpose. The aim is to analyze the features of aortic valve allografts decellularized with trypsin solution or a combination of the sodium dodecyl sulfate/deoxycholate solution.Materials and methods. We performed histological, bacteriological and mechanical testing of graft.Results. It was found that after the removal of cellular elements with trypsin the strength of allograft significantly decreased compared with the control group and the samples exposed to detergent treatment. Histological analysis of aortic allograft specimens showed signifi cantly impaired spatial organization of the connective tissue framework of the valve after the enzymatic treatment.Conclusion. Detergent decellularization is the preferred method for obtaining cell-free aortic allografts because it can effectively remove the donor cells and maintain the microstructure and biomechanical properties of the valve graft. Цель. Анализ особенности аллографтов аортального клапана, децеллюляризованных с помощью раствора трипсина или комбинированного действия растворов додецилсульфата и дезоксихолата натрия.Материалы и методы. Проводили гистологические, бактериологические и мехнические испытания графта.Результаты. После удаления клеточных элементов трипсином достоверно снижалась прочность аллографта по сравнению с группой контроля и образцов после детергентной обработки. Гистологический анализ образцов аортальных аллографтов достоверно показал выраженное нарушение пространственной организации соединительнотканного каркаса клапана после ферментной обработки.Заключение. Детергентная децеллюляризация является предпочтительным методом получения бесклеточного аортального аллографта, т. к. позволяет эффективно удалить клетки донора и сохранить микроструктуру и биомеханические свойства графта.

APPLICATION OF SIX-COLOR FLOW CYTOMETRIC ANALYSIS FOR IMMUNE PROFILE MONITORING

The article deals with applications of six-color flow analysis for studying the immune state parameters by means of flow cytometry. Whole blood from healthy donors was stained with combination of monoclonal antibodies, i.e., HLA-DR-FITC, CD16+56-PE, CD4-ECD, CD19-ECD, CD8-PC5.5, CD3-PC7, CD45-APC (Beckman Coulter, USA) using a “no-wash” technology. To adjust the photomultiplier (PMT) voltage, we used the tubes with each of the tested monoclonal antibodies, PMT voltage was considered optimal when the negative population was located in the middle of the first decade at the logarithmic scale. The compensatory adjustment was performed in automatic mode, using Navios software (Beckman Coulter, USA). We discuss an optimal gating strategy in order to assess the populations of interest: total T cells (CD3+CD19-), T helper cells (CD3+CD4+), cytotoxic T lymphocytes (CD3+CD8+), B cells (CD3-CD19+), NK cells (CD3-CD16+CD56+), NKT cells (CD3+CD16+CD56+), activated T lymphocytes (CD3+HLA-DR+)

PLASMINOGEN ACTIVATOR INHIBITOR GENE POLYMORPHISM IN EVALUATION OF THE VARIOUS LOCALIZATION THROMBOSIS RISKS (PILOT STUDY)

Aim. Assessment of the impact of genetic polymorphism of rs1799889 gene of PAI-1 inhypofibrinolytic state development, with the inclusion of the protein product amount shifts, in patients with thrombosis of different localizations.Material and methods. Totally 50 patients studied with thrombosis of different localization and etiology in anamnesis, and 25 controls — almost healthy donors, living on theterritoryofNovosibirskcity and the region. All patients underwent genotyping of polymorphism -675 4G/5G of the PAI-1 gene, and concentration of plasminogen activator inhibitor (PAI1) in blood plasma measurement.Results. In the groups of patients the prevalence studied of the gene PAI-1 variants -675 4G/5G, and the level of PAI1 measured, the influence of genotypes on the level of protein assessed. Assessment of allele variant 4G and high level of PAI1 is highly informative for the estimation of thrombosis risk.Conclusion. Genotypes 4G/4G and 5G/4G of gene PAI1, together with increased level of PAI1 are diagnostically important markers of endothelium dysfunction, hypo fibrinolysis condition and hence are predictors of clotting

THE “PROTOCOL” STUDY: PRELIMINARY RESULTS

Aim. To reveal the association of gene polymorphism CYP2С19*2 and recurrent early stent thrombosis in coronary vessels, and paradoxal response to clopidogrel intake in patients — the inhabitants of Siberian Region — after acute coronary syndrome.Material and methods. Totally 105 patients studied, hospitalized for stenting of coronary arteries in acute coronary syndrome, never received clopidogrel previously. The polymorphism of studied СУР2С19: *2, *3, *17 alleles, as assessment of platelet aggregation with ADP before and after clopidogrel intake, as of endpoints on safety and efficacy during 30 days (thrombotic complications, bleeding).Results. Within the selected patients there was no any significant association of any CYP2С19*2 and/or CYP2С19*3 alleles and paradoxic laboratory reaction. There was significant association of the CYP2C19*17 gene carriage and bleedings. When comparing the groups of patients having or not having complications related to clopidogrel (thrombotic or bleedings), there was significant difference in residual platelet aggregation.Conclusion. The results of the study might be strongly significant for the decisions making on double antiplatelet therapy and on the tactics of drugs preference