Hemolysis and ATP release from human and rat erythrocytes under conditions of hypoxia: a comparative study

Red blood cells are involved not only in transportation of oxygen and carbon dioxide but also in autoregulation of vascular tone by ATP release in hypoxic conditions. Molecular mechanisms of the ATP release from red blood cells in response to a decrease in partial oxygen pressure still remain to be elucidated. In this work we have studied effects of hypoxia on red blood cell hemolysis in humans and rats and compared the effects of inhibitors of ecto-ATPase and pannexin on the release of ATP and hemoglobin from rat erythrocytes. The 20-min hypoxia at 37°C increased hemolysis of red blood cells in humans and rats 1.5- and 2.5-fold, respectively. In rat erythrocytes a significant increase in hypoxia-induced extracellular ATP level was found only in the presence of ecto-ATPase inhibitor ARL 67156. In these conditions we observed a positive correlation (R2 = 0.5003) between the increase in free hemoglobin concentration and the ATP release. Neither carbenoxolon nor probenecid, the inhibitors of low-selectivity pannexin channels, altered the hypoxia-induced ATP release from rat erythrocytes. The obtained results indicate a key role of hemolysis in the ATP release from red blood cells

Deoxygenation affects composition of membrane-bound proteins in human erythrocytes

Background/Aims: ATP release from erythrocyte plays a key role in hypoxia-induced elevation of blood flow in systematic circulation. We have previously shown that hemolysis contributes to erythrocyte ATP release triggered by several stimuli, including hypoxia, but the molecular mechanisms of hypoxia-increased membrane fragility remain unknown. Methods: In this study, we compared the action of hypoxia on hemolysis, ATP release and the composition of membrane-bound proteins in human erythrocytes. Results: Twenty minutes incubation of human erythrocytes in the oxygen-free environment increased the content of extracellular hemoglobin by ∼1.5 fold. Paired measurements of hemoglobin and ATP content in the same samples, showed a positive correlation between hemolysis and ATP release. Comparative analysis of SDS-PAGE electrophoresis of erythrocyte ghosts obtained under control and deoxygenated conditions revealed a ∼2-fold elevation of the content of membrane-bound protein with Mr of ∼60 kDa. Conclusion: Deoxygenation of human erythrocytes affects composition of membrane-bound proteins. Additional experiments should be performed to identify the molecular origin of 60 kDa protein and its role in the attenuation of erythrocyte integrity and ATP release in hypoxic conditions

Опыт вакцинации 13-валентной конъюгированной пневмококковой вакциной пациентки с ювенильным идиопатическим артритом с частыми респираторными инфекциями на фоне терапии метотрексатом

The article presents the experience of vaccination with a pneumococcal 13-valent conjugate vaccine (PCV13) of a patient aged 5 years with oligoarticular juvenile idiopathic arthritis (JIA) receiving methotrexate at a dose of 15 mg/m2 per week subcutaneously. Treatment with methotrexate provided a remission of JIA, but was accompanied by frequent respiratory infections — up to 8 times a year. During infection progression, methotrexate injections were omitted. Gaps in the treatment with methotrexate were accompanied by an exacerbation of the underlying condition. Vaccination of the patient with PCV13 reduced the frequency of respiratory infections to 2 times a year, which was accompanied by the development of persistent remission of the disease. Adverse events and exacerbation of JIA in a child after vaccination with PCV13 were not registered.Представлен опыт вакцинации 13-валентной конъюгированной пневмококковой вакциной (13ПКВ) пациентки в возрасте 5 лет с олигоартикулярным ювенильным идиопатическим артритом (ЮИА), получавшей метотрексат в дозе 15 мг/м2 в неделю подкожно. Лечение метотрексатом обеспечило ремиссию ЮИА, но сопровождалось частыми респираторными инфекциями — до 8 раз в год. При развитии инфекций инъекции метотрексата пропускались. Перерывы в лечении метотрексатом сопровождались обострением основного заболевания. Вакцинация пациентки 13ПКВ обеспечила снижение частоты респираторных инфекций до 2 раз в год, что сопровождалось развитием стойкой ремиссии заболевания. Нежелательных явлений и обострения ЮИА у ребенка на фоне вакцинации 13ПКВ не зарегистрировано

Nature-aligned Education Technologies as the Basis of Forming an Individual Noospheric Consciousness

With the aim of identifying the mechanisms of the emergence of positive changes in the formation of students’ consciousness, factors determining natural conformity of perceiving reality are considered. A number of features of shifting from sinistrocerebral to holistic thinking are considered. It is shown that the introduction of noospheric consciousness into the realization of individual basic functions displays his/her ability to self-awareness of an integrated system of structural-functional dynamics of the psyche. The problem of bringing the education system into line with the major trends in the development of scientific research is set up. In this regard, a mental image in the shape of a holographic form is considered as a holistic holographic image of an event, phenomenon, and subject filled with information and perceived by all senses individually. It is shown that consideration of dynamics of forming mental images in educational processes is a factor of ensuring psychological health. It is shown that nature-aligned consciousness is emergent in the organization of cognition and comprehension, when it unexpectedly arises from the gross properties of consciousness and is projected onto each individual consciousness able to engage in a global neural network – the noosphere, the information layer, the collective unconscious and superconscious. It is shown that instead of a trained sinistrocerebral, clip, terminal way of thinking, when human capabilities are limited and health is exhausted, a nature-aligned, holistic, neural network, emergent way of thinking is possible that opens up new possibilities of the pupil, and, most importantly, saves health. Such feature of the brain as its permanent activity is considered, which is presented by specific cerebral oscillations of the brain rhythms: delta rhythms (0.5 to 4 Hz), theta rhythms (4–8 Hz), alpha rhythms (8–13 Hz), beta rhythms (14–40 Hz) and gamma rhythms (over 40 Hz) and sigma-rhythms (10–16 Hz from 13 to 14 cycles per second). Each of the rhythms is a symptom of a certain state of consciousness. The importance of changing the mode of operation of the brain and nervous system is shown: activity – relaxation in the learning process. It is discussed that the modern system of education and personal development faces the task, the solution of which noospheric psycho-pedagogy offers: how to shape nature-aligned consciousness of the individual, which allows discovering unlimited resources of a person’s alpha theta, sigma and gamma states. It is argued that in the light of the latest scientific discoveries on the modern stage of society development the teacher should know and use in their activities to nature-aligned educational technologies, aimed at forming nature-aligned noospheric consciousness of the individual. A teacher can and should contribute to the harmonious work of a brain of the student in the learning process. This is possible if the teacher applies bioadequate methods of teaching and uses bioadequate textbooks. It is recommended for the teacher to take a refresher course “Holistic thinking” and “Nature-aligned methods of teaching academic disciplines.

Flora and Vegetation of the Aleksandrovskiy Wildlife Area (Tyumen Oblast, Vikulovo District)

Участок «Александровский», зарезервированный под организацию заказника, расположен в подтайге в восточной части Викуловского района Тюменской области и занимает площадь 95830,2 га. Флористические исследования на данной территории в связи с ее труднодоступностью – высоким уровнем заболоченности и отсутствием дорог – носят фрагментарный характер. Цель работы – получение актуальных данных о состоянии флоры и растительности, наличии видов растений и растительных сообществ, нуждающихся в охране. В статье описываются основные типы фитоценозов участка «Александровский»: мелколиственные, светлохвойные и широколиственные леса, ивняки, злаково-разнотравные суходольные луга и осоково-тростниковые займища в сочетании с галофитными сообществами, сосново-кустарничково- сфагновые болота – рямы, а также прибрежно-водная растительность. Преобладают на обследованной территории березовые, осиновые и осиново-березовые орляковые, разнотравные и злаково-разнотравные леса. Флористическое разнообразие исследуемого участка представлено 410 видами сосудистых растений, в числе которых 80 отмечены впервые для Викуловского района. Выявлены новые местонахождения 10 видов сосудистых растений, по одному виду мхов и лишайников, включенных в Красную книгу Тюменской области, в том числе Cypripedium calceolus L., Lobaria pulmonaria (L.) Hoffm., занесенных в Красную книгу Российской Федерации. Отмечены 14 видов, встречающихся относительно редко на территории Тюменской области. Для каждого вида указываются координаты, географические пункты сбора, местообитания, даты сборов, для «краснокнижных» видов – категория редкости и состояние ценопопуляций. Полученные данные позволяют уточнить местонахождение видов и границы их ареалов на территории ЗауральяThe Aleksandrovskiy wildlife area (95830.2 ha) reserved for a sanctuary is located in sub-taiga of the western part of Vikulovo District, Tyumen Oblast. A complete floral survey of the area is unattainable at present because it is a vast wetland with very few roads. The study is aimed at evaluating the current state of local flora and vegetation, and identifying plants and plant communities that require protection. Main types of phytocenoses in the Aleksandrovskiy wildlife area were described, which include small-leaved, light-coniferous and broad-leaved forests, willow coppices, cereal-forb dry meadows, sedge-reed floodplain meadows mixed with halophyte communities, pine-subshrub-sphagnum raised bogs (riams), and semi-aquatic vegetation. Birch, aspen and aspen-birch bracken, forb and cereal-forb meadows predominate in the surveyed area. The floral diversity of the site amounts to 410 species of vascular plants, with 80 found in Vikulovo District for the first time. New sites for 10 vascular plants, one moss and one lichen species included in Red Data Book of Tyumen Oblast were found, among them Cypripedium calceolus L., Lobaria pulmonaria (L.) Hoffm. included in Red Data Book of the Russian Federation. Fourteen species which are relatively rare in Tyumen Oblast were registered. All found species were described with a reference to their habitats, geographical coordinates of the collection sites and the date of collection. Additionally, the Red Data Book species description included their rarity status and the condition of the coenopopulations. The results allow to define the species distribution areas in Transurali

Proteomics-based identification of hypoxia-sensitive membrane-bound proteins in rat erythrocytes

This study examines the action of hypoxia on integrity, fluidity and protein composition of red blood cell (RBC) membrane. Twenty-min exposure to oxygen-free environment decreases rat RBC integrity documented by 3-fold elevation of hemoglobin release without any action on the membrane fluidity estimated by electron magnetic resonance spectroscopy of spin-labeled stearic acid analogues. The proteomics technology in combination with relative label free quantification analysis revealed a dozen of membrane-bound proteins, including elevated content of hemoglobin, reproducibly affected by hypoxia. Mapping the identified proteins in the KEGG pathway database we found that the proteins of multi subunit Cullin-Rbx E3 ubiquitin ligase complex are presented in normoxic RBC ghosts but not in the hypoxic samples. Our results suggest that Cullin-Rbx E3 complex, associated with RBC membrane in normoxia, provides detection and deletion of membrane proteins damaged by reactive oxygen species. In hypoxic conditions, deoxy-Hb binds to band 3 protein, resulting in dissociation of Cullin-Rbx E3 complex from RBC membrane and impaired clearance of damaged cytoskeleton proteins. These rearrangements of membrane proteins might be involved in attenuated membrane integrity revealed in hypoxic RBC

Proteomics-based identification of hypoxia-sensitive membrane-bound proteins in rat erythrocytes

This study examines the action of hypoxia on integrity, fluidity and protein composition of red blood cell (RBC) membrane. Twenty-min exposure to oxygen-free environment decreases rat RBC integrity documented by 3-fold elevation of hemoglobin release without any action on the membrane fluidity estimated by electron magnetic resonance spectroscopy of spin-labeled stearic acid analogues. The proteomics technology in combination with relative label free quantification analysis revealed a dozen of membrane-bound proteins, including elevated content of hemoglobin, reproducibly affected by hypoxia. Mapping the identified proteins in the KEGG pathway database we found that the proteins of multi subunit Cullin-Rbx E3 ubiquitin ligase complex are presented in normoxic RBC ghosts but not in the hypoxic samples. Our results suggest that Cullin-Rbx E3 complex, associated with RBC membrane in normoxia, provides detection and deletion of membrane proteins damaged by reactive oxygen species. In hypoxic conditions, deoxy-Hb binds to band 3 protein, resulting in dissociation of Cullin-Rbx E3 complex from RBC membrane and impaired clearance of damaged cytoskeleton proteins. These rearrangements of membrane proteins might be involved in attenuated membrane integrity revealed in hypoxic RBC

Hemolysis and ATP release from human and rat erythrocytes under conditions of hypoxia: a comparative study

Red blood cells are involved not only in transportation of oxygen and carbon dioxide but also in autoregulation of vascular tone by ATP release in hypoxic conditions. Molecular mechanisms of the ATP release from red blood cells in response to a decrease in partial oxygen pressure still remain to be elucidated. In this work we have studied effects of hypoxia on red blood cell hemolysis in humans and rats and compared the effects of inhibitors of ecto-ATPase and pannexin on the release of ATP and hemoglobin from rat erythrocytes. The 20-min hypoxia at 37°C increased hemolysis of red blood cells in humans and rats 1.5- and 2.5-fold, respectively. In rat erythrocytes a significant increase in hypoxia-induced extracellular ATP level was found only in the presence of ecto-ATPase inhibitor ARL 67156. In these conditions we observed a positive correlation (R2 = 0.5003) between the increase in free hemoglobin concentration and the ATP release. Neither carbenoxolon nor probenecid, the inhibitors of low-selectivity pannexin channels, altered the hypoxia-induced ATP release from rat erythrocytes. The obtained results indicate a key role of hemolysis in the ATP release from red blood cells

Deoxygenation affects composition of membrane-bound proteins in human erythrocytes

Background/Aims: ATP release from erythrocyte plays a key role in hypoxia-induced elevation of blood flow in systematic circulation. We have previously shown that hemolysis contributes to erythrocyte ATP release triggered by several stimuli, including hypoxia, but the molecular mechanisms of hypoxia-increased membrane fragility remain unknown. Methods: In this study, we compared the action of hypoxia on hemolysis, ATP release and the composition of membrane-bound proteins in human erythrocytes. Results: Twenty minutes incubation of human erythrocytes in the oxygen-free environment increased the content of extracellular hemoglobin by ∼1.5 fold. Paired measurements of hemoglobin and ATP content in the same samples, showed a positive correlation between hemolysis and ATP release. Comparative analysis of SDS-PAGE electrophoresis of erythrocyte ghosts obtained under control and deoxygenated conditions revealed a ∼2-fold elevation of the content of membrane-bound protein with Mr of ∼60 kDa. Conclusion: Deoxygenation of human erythrocytes affects composition of membrane-bound proteins. Additional experiments should be performed to identify the molecular origin of 60 kDa protein and its role in the attenuation of erythrocyte integrity and ATP release in hypoxic conditions

Pneumococcal Vaccine in Patients with Systemic Juvenile Idiopathic Arthritis Receiving Biologic Therapy: International Practice Review

International practice of immunization against pneumococcus in patients with systemic juvenile idiopathic arthritis (SJIA) receiving biological therapy is generalized in this review. High efficiency and safety of pneumococcal vaccines in children with SJIA is presented. Numerous researches show the adequate immune response after vaccination as well as alongside with genetically engineered biologic drugs therapy. Prevention of pneumococcal disease in patients with SJIA reduces the risk of development of pneumococcal diseases severe complications