The development of adequate finite element models of load-bearing structures coating “Vnukovo-1 airport complex” to solve the problems of monitoring their technical condition

This article considers the issues of designing adequate finite element models of weight-bearing structures based on the roof structures of the Vnukovo-1 airport, which is made in the form of a spatial core structure. Additional actions on the covering structure model in terms of the strength checking calculation procedure are presented, its purpose is to make the adequate model for its further use in the design of structural health monitoring. The article presents the results of modeling, comparative research with the data of nature experiment

ОСОБЕННОСТИ ПРОДУКЦИИ ВНЕКЛЕТОЧНЫХ ВЕЗИКУЛ КЛЕТКАМИ ЛИНИИ THP-1 ПРИ СТИМУЛЯЦИИ IN VITRO

HighlightsExtracellular vesicles (EVs) are membrane objects with inner and surface molecular cargo performing various biological functions associated with intercellular connection. Therefore, they are a promising object for diagnostic and therapeutic purposes.THP-1 is a human leukemia monocytic cell line and a classic model for monocyte/macrophage function and differentiation. Spectrum of THP-1-derived extracellular vesicles are heterogenic and are not to date totally clarified.The comparison of EVs produced by stimulated THP-1 cells is performed in the study. Aim. Quantitatively and qualitatively characterize THP-1-derived extracellular vesicles in concordance on type and dose of used stimulators.Methods. THP-1 cells were stimulated by PMA (4-phorbol 12-myristate 13-acetate), LPS (lipopolysaccharide) and TNF (tumor necrosis factor). Cell activation and viability were assessed by the expression of CD54 and DAPI (4′,6-diamidino-2-phenylindole) staining respectively. Size and quantitative characteristics were assessed using nanoparticle tracking analysis (NTA). Qualitative assessment of surface molecular markers was performed by high-sensitivity multicolor flow cytometry (hs-FCM).Results. TNF and PMA stimulation in medium and high doses led to an increase in quantity of small EVs (30–150 nm) that express tetraspanins CD9 and CD63. Low doses of TNF and PMA did not lead to a significant increase of EVs compared to control. There was a decrease in amount of small EVs (30–150 nm) in LPS-stimulated probes. However, medium and high doses of LPS stimulated selective CD63-positive EVs secretion with absence of CD9-positive events. Cell activation assessed by CD54 expression have been dose-dependent and was most prominent under PMA stimulation.Conclusion. Every type of THP-1 cell activators promotes changes in the functional activity of cells towards production of extracellular vesicles. Moreover, we demonstrate quantitative and qualitative difference in THP-1-derived vesicles under different stimulation conditions. Medium and high doses of PMA and TNF were the most relevant in stimulation of vesiculation. LPS-stimulation could be used to selectively obtain CD63-positive fraction of EV.Основные положенияВнеклеточные везикулы (ВВ) – это мембранные объекты c молекулярным грузом внутри себя и на поверхности, выполняющие разнообразные биологические функции, связанные с межклеточным взаимодействием, поэтому являются перспективным объектом для диагностических и терапевтических целей.Культура клеток острого миелоидного лейкоза человека – THP-1 – классическая модельная моноцитарно-макрофагальная система. Спектр секретируемых этой культурой ВВ гетерогенен и недостаточно охарактеризован к настоящему времени.В работе проводится сравнение продуцируемых клетками THP-1 ВВ при воздействии различных стимуляторов. Цель. Дать количественную и качественную характеристику внеклеточных везикул, продуцируемых клетками линии THP-1, в зависимости от использованного стимулятора и его дозы.Материалы и методы. В исследовании были использованы клетки культуры THP-1. Для стимуляции клеток использовались: PMA (4-форбол-12-миристат-13-ацетат) в дозах 5, 16 и 50 нг/мл, ЛПС (липополисахарид) в дозах 0,1 нг/мл, 0,3 нг/мл, 1,0 нг/мл и ФНО (фактор некроза опухоли) в дозах 5, 10 и 20 нг/мл. Активация клеток оценивалась изменением уровня экспрессии CD54, а наличие апоптоза/некроза – посредством подсчета DAPI (4′,6-диамидино-2-фенилиндол)-позитивных клеток с помощью высокочувствительной проточной цитометрии (hs-FCM). Размерность продуцируемых везикул и их количественная характеристика была дана с использованием анализа траектории наночастиц (NTA). Качественная характеристика экспрессируемых поверхностных молекулярных маркеров была дана с помощью высокочувствительной многоцветной проточной цитометрии (hs-FCM).      Результаты. Стимуляция ФНО и PMA в средних и высоких дозировках приводит к нарастанию секретируемой фракции малых (30–150 нм) внеклеточных везикул, экспрессирующих тетраспаниновые маркеры (CD9, CD63). Малые дозы ФНО и PMA не приводили к достоверному приросту количества ВВ. При использовании всех дозировок ЛПС отмечалось некоторое количественное снижение ВВ в диапазоне 30-150 нм (малые ВВ). Однако средние и высокие дозы ЛПС вызывали достоверную селективную секрецию CD63-положительных ВВ при полном отсутствии динамики изменения CD9-позитивных везикул. Активация клеток, оцениваемая по экспрессии CD54, носила дозозависимый характер и была наиболее выражена при стимуляции PMA.Заключение. Каждый вид использованной стимуляции клеток THP-1 провоцирует изменение функциональной активности клеток в отношении продуцируемых ими внеклеточных везикул. При этом может меняться не только количественный, но и качественный состав продуцируемых объектов. Наиболее предпочтительными в стимуляции везикуляции оказались средние и высокие дозы ФНО и PMA. Стимуляция ЛПС в средних и высоких дозах может быть использована для получения изолированной CD63-положительной фракции ВВ

The development of adequate finite element models of load-bearing structures coating “Vnukovo-1 airport complex” to solve the problems of monitoring their technical condition

This article considers the issues of designing adequate finite element models of weight-bearing structures based on the roof structures of the Vnukovo-1 airport, which is made in the form of a spatial core structure. Additional actions on the covering structure model in terms of the strength checking calculation procedure are presented, its purpose is to make the adequate model for its further use in the design of structural health monitoring. The article presents the results of modeling, comparative research with the data of nature experiment

Deep Immunophenotyping of Circulating T and B Cells in Relapsing Adult-Onset Still’s Disease

Adult-onset Still’s disease (AOSD) is a complex systemic inflammatory disorder, categorized as an ‘IL-1 driven’ inflammasomapathy. Despite this, the interaction between T and B cells remains poorly understood. We conducted a study, enrolling 7 patients with relapsing AOSD and 15 healthy control subjects, utilizing deep flow cytometry analysis to examine peripheral blood T- and B-cell subsets. T-cell and B-cell subsets were significantly altered in patients with AOSD. Within CD4+ T cells, Th2 cells were decreased. Additionally, Th17 cell and follicular Th cell subsets were altered within CD45RA–CD62L+ and CD45RA–CD62L– Th cells in patients with AOSD compared to healthy controls. We identified changes in CD8+ T cell maturation and ‘polarization’ in AOSD patients, with an elevated presence of the TEMRA CD8+ T cell subset. Furthermore, the percentage of Tc1 cells was decreased, while the frequency of CCR6–CXCR3– Tc2 cells was elevated. Finally, we determined that the frequency of CD5+CD27– B cells was dramatically decreased in patients with AOSD compared to healthy controls. Further investigations on a large group of patients with AOSD are required to evaluate these adaptive immunity cells in the disease pathogenesis

Dysregulated Immune Responses in SARS-CoV-2-Infected Patients: A Comprehensive Overview

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first detected in humans more than two years ago and caused an unprecedented socio-economic burden on all countries around the world. Since then, numerous studies have attempted to identify various mechanisms involved in the alterations of innate and adaptive immunity in COVID-19 patients, with the ultimate goal of finding ways to correct pathological changes and improve disease outcomes. State-of-the-art research methods made it possible to establish precise molecular mechanisms which the new virus uses to trigger multisystem inflammatory syndrome and evade host antiviral immune responses. In this review, we present a comprehensive analysis of published data that provide insight into pathological changes in T and B cell subsets and their phenotypes, accompanying the acute phase of the SARS-CoV-2 infection. This knowledge might help reveal new biomarkers that can be utilized to recognize case severity early as well as to provide additional objective information on the effective formation of SARS-CoV-2-specific immunity and predict long-term complications of COVID-19, including a large variety of symptoms termed the ‘post-COVID-19 syndrome’