Structural characterization of platelets and platelet microvesicles

© 2016, Pleiades Publishing, Ltd.Platelets are blood cells without nuclei, which, in conjunction with fibrin, cause bleeding to stop (hemostasis). Cellular microvesicles are microscopic particles released into extracellular space under activation and/or apoptosis of cells of different types. Platelet microvesicles form the main population of blood circulating through microvesicles and play an important role in the reactions of hemostasis, thrombosis, and many other (patho)physiological processes. Despite the large number of studies that have been devoted to the function of platelet microvesicles, the mechanisms of their formation and structural details remain poorly understood. The ultrastructure of the initial platelets and microvesicles formed in vitro from resting cells and platelets activated by arachidonic acid, ADP, thrombin, and calcium ionophore A23187 is investigated in this study. The intracellular origin, stages of formation, structural diversity, and size of microvesicles were analyzed according to the results of transmission electron microscopy of human platelets and isolated microvesicles. It was shown that thrombin, unlike other activators, not only stimulates microvesiculation of the plasma membrane, but also causes decomposition of cells with the formation of subcellular particles that have sizes comparable with the size of the microvesicles from the outer membrane of the cells. Some of these microparticles are cellular organelles surrounded by a thin membrane. The size of isolated microvesicles ranges from 30 to 500 nm, but their size distribution depends on the nature of the activating stimulus. The obtained results contain new data on the formation of platelet microvesicles and their structural diversity, which are important for understanding of their multiple functions in health and disease

Effects of succinate on the growth rates of potato, Rauwolfia, and ginseng in vitro

Effects of succinate on in vitro potato reproduction, callus cultures of Rauwolfia serpentina, and Panax ginseng roots were studied. The optimum succinate concentration giving the maximum the growth rate of potato shoots and callus mass, was found to be 10-5 M. Succinate treatment increased the absolute yield of the alkaloid ajmaline in Rauwolfia callus culture, in spite of a decrease in its specific content

Effects of Succinate on the Growth Rates of Potato, Rauwolfia, and Ginseng in vitro

Effects of succinate on in vitro potato reproduction, callus cultures of Rauwolfia serpentina, and Panax ginseng roots was studied. The optimum succinate concentration giving the maximum the growth rate of potato shoots and callus mass, was found to be 10-5 M. Succinate treatment increased the absolute yield of the alkaloid ajmaline in Rauwolfia callus culture, in spite of a decrease in its specific content

Analysis of the experience of implementing urban redevelopment projects in Russia

The relevance of the study is determined by the identified shortage of the urban environment in Russia, namely those types of buildings that meet modern requirements. In large cities, the only way to change this situation is redevelopment. This article considers options for stimulating the development of urban areas and optimization of their use through redevelopment. The definitions of the concepts of redevelopment, renovation, gentrification, revitalization are delimited. The classification of types of programs according to the level of management is performed. The advantages and disadvantages of the redevelopment process are presented. Based on the analysis of the regulatory and legal framework, the authors revealed that the concept of “redevelopment” is not defined in any way in it. The practical experience of implementing redevelopment projects and programs in Russia, including development projects of the main industrial zones of the city of Moscow, which are inextricably linked with the strategic development of the constituent entities of the Russian Federation and the implementation of national projects, is analysed. Special attention is paid to the redevelopment project management issues, proposals are presented to increase the efficiency of program management based on the Japanese P2M project management standard. At the end of the study, the authors revealed the results achieved in the implementation of redevelopment projects and evaluated their effectiveness in terms of various indicators

Intracellular origin and ultrastructure of platelet-derived microparticles

© 2017 International Society on Thrombosis and Haemostasis Essentials Platelet microparticles play a major role in pathologies, including hemostasis and thrombosis. Platelet microparticles have been analyzed and classified based on their ultrastructure. The structure and intracellular origin of microparticles depend on the cell-activating stimulus. Thrombin-treated platelets fall apart and form microparticles that contain cellular organelles. Summary: Background Platelet-derived microparticles comprise the major population of circulating blood microparticles that play an important role in hemostasis and thrombosis. Despite numerous studies on the (patho)physiological roles of platelet-derived microparticles, mechanisms of their formation and structural details remain largely unknown. Objectives Here we studied the formation, ultrastructure and composition of platelet-derived microparticles from isolated human platelets, either quiescent or stimulated with one of the following activators: arachidonic acid, ADP, collagen, thrombin or calcium ionophore A23187. Methods Using flow cytometry, transmission and scanning electron microscopy, we analyzed the intracellular origin, structural diversity and size distributions of the subcellular particles released from platelets. Results The structure, dimensions and intracellular origin of microparticles depend on the cell-activating stimulus. The main structural groups include a vesicle surrounded by one thin membrane or multivesicular structures. Thrombin, unlike other stimuli, induced formation of microparticles not only from the platelet plasma membrane and cytoplasm but also from intracellular structures. A fraction of these vesicular particles having an intracellular origin contained organelles, such as mitochondria, glycogen granules and vacuoles. The size of platelet-derived microparticles depended on the nature of the cell-activating stimulus. Conclusion The results obtained provide a structural basis for the qualitative differences of various platelet activators, for specific physiological and pathological effects of microparticles, and for development of advanced assays

Применение ингибиторов фактора некроза опухоли α у женщин с анкилозирующим спондилитом во время беременности

Objective: to present our own experience of tumor necrosis factor α (TNFα) inhibitors (iTNFα) usage during pregnancy in women with ankylosing spondylitis (AS), to assess AS activity and outcomes of gestation.Patients and methods. A prospective observation of 55 pregnant women with AS who met the modified New York criteria of 1984. Fifty-six pregnancies were followed. The average age of the patients was 31.7±4.7 years, the duration of the disease was 132.2±85.4 months. The median BASDAI for pregnancy trimesters was 2.4 [1.2; 4.4], 2.7 [1.4; 4.2] and 2.2 [1.5; 4.0], respectively. 14 women received iTNFα 3 months before pregnancy.Results and discussion. In the first trimester, TNFα was used in 9 (16.1%) patients, in the second – in 9 (16.1%) and in the third – in 5 (9.3%); the median BASDAI for trimesters was 2.3 [1.0; 3.7], 3.4 [1.2; 3.5], 3.0 [0.8; 3.4], respectively. All patients who discontinued iTNFα just before or in early pregnancy had indications for resuming therapy in the second half of gestation. Cancellation of iTNFα at the end of the second trimester was not a risk factor for high activity in the third trimester. There was 1 adverse pregnancy outcome. In other cases, childbirth occurred at 38.9±1.4 weeks, newborns' body weight was 3273.1±435.6 g.Conclusion. Women with AS who plan a pregnancy should be prescribed drugs with the maximum allowed duration of use during gestation. Cancellation of iTNFα before and in early pregnancy is a risk factor for high AS activity, while renewal of iTNFα therapy during pregnancy is not always effective.Цель исследования – представить собственный опыт применения ингибиторов фактора некроза опухоли (иФНОα) во время беременности у женщин с анкилозирующим спондилитом (АС), оценить активность АС и исходы гестации.Пациенты и методы. Проведено проспективное наблюдение 55 беременных с АС, соответствовавших модифицированным Нью-Йоркским критериям 1984 г. Прослежено 56 беременностей. Средний возраст пациенток составлял 31,7±4,7 года, продолжительность болезни – 132,2±85,4 мес. Медиана BASDAI по триместрам беременности – 2,4 [1,2; 4,4], 2,7 [1,4; 4,2] и 2,2 [1,5; 4,0] соответственно. ИФНОα за 3 мес до наступления беременности получали 14 женщин.Результаты и обсуждение. В I триместре иФНОα применяли 9 (16,1%) пациенток, во II – 9 (16,1%) и в III – 5 (9,3%); медиана BASDAI по триместрам составляла 2,3 [1,0; 3,7], 3,4 [1,2; 3,5], 3,0 [0,8; 3,4] соответственно. Все пациентки, отменившие иФНОα накануне или на ранних сроках беременности, имели показания к возобновлению терапии во второй половине гестации. Отмена иФНОα в конце II триместра не являлась фактором риска высокой активности в III триместре. Имел место 1 неблагоприятный исход беременности. В остальных случаях роды произошли на сроке 38,9±1,4 нед, масса тела новорожденных – 3273,1±435,6 г.Заключение. Женщинам с АС, планирующим беременность, необходимо назначать препараты с максимально разрешенным сроком применения во время гестации. Отмена иФНОα перед беременностью и на ее ранних сроках – фактор риска высокой активности АС, при этом возобновление терапии иФНОα во время беременности не всегда эффективно

MODY caused by a mutation in the insulin gene

MODY10 is a rare subtype of MODY diabetes, which caused by heterozygous mutations in the insulin gene INS. There are single descriptions of families with MODY-INS or MODY10 in the literature, its clinical course is not well understood. We present a case of MODY10 in a boy with a history of diabetes mellitus (DM) in three generations (father and paternal grandmother). Proband was diagnosed with diabetes mellitus at the age of 7 years. The glycaemia at the onset of the diabetes was 10.2 mmol/l, HbA1c — 7.6%, islet cell autoantibodies (ICA), insulin autoantibodies (IAA), glutamic acid decarboxylase antibodies (GADA) and islet tyrosine phosphatase 2 (IA2) antibodies (IA2) were not detected. According to the results of the oral glucose tolerance test, fasting blood glucose was 6.5 mmol/l, in 120 minutes 10.3 mmol/l, which corresponded to the diagnosis of impaired glucose tolerance. Diet with restriction of easily digestible carbohydrates was recommended, than gliclazide was added to the therapy, which the proband received for 3 years. At the age of 10, a deterioration in the parameters of carbohydrate metabolism was noted, which insulin therapy was added. Examination at the age of 12 revealed a decrease in C-peptide secretion. The child’s father has a similar phenotype — slowly progressive disorders of carbohydrate metabolism from 6 years old, from 10 years old — insulin therapy. A genetic test was provided, in the child and his father was detected a previously undescribed heterozygous mutation in the INS p.C31W. Thus, in our clinical case, MODY10 was characterized by a milder course than T1DM, but eventually leading to the development of insulin demand, which distinguishes it from the most common forms of MODY. Currently, there is no specific therapy, and the detection of a mutation in the INS gene does not affect therapeutic tactics, however, a correct genetic diagnosis makes it possible to predict the course of diabetes and provide genetic counseling to the family

Detection of macro-thyrotropinaemia in patients with Hashimotos thyroiditis and subclinical hypothyroidism

The level of thyroid stimulating hormone is one of the diagnostic indicators of thyroid function. In subclinical hypothyroidism, its concentration in the blood serum increases, while the level of thyroid hormones remains normal. One of the reasons for this is the phenomenon of macrotyrotropinemia, in which the macro isoforms of thyrotropin (a complex of thyrotropic hormone with immunoglobulin) are present in the blood. It is assumed that the biological activity of macrotyrotropin is low, and may accumulate in the circulation, causing a falsely elevated level of thyroid-stimulating hormone in serum. The aim of this study is to identify the nature and prevalence of the macrothyrotropinemia phenomenon among patients with subclinical hypothyroidism in presence of autoimmune thyroiditis and a group of healthy donors. Materials and methods: Fifty serum samples of venous blood served as the material for the study: 30 patients with subclinical hypothyroidism in presence of autoimmune thyroiditis; 10, with manifesting hypothyroidism, 10 conditionally healthy donors without thyroid gland pathology (control group). The group was derived from results of the clinical laboratory at the Clinical Hospital at the Kazan station railway. Patients’ blood serum was screened for the presence of macrotrorotropin by polyethylene glycol precipitation method, followed by analysis by gel filtration chromatography. Results of this study were as follows: screening of blood sera was performed by gel filtration and affinity chromatography. Polyethylene glycol was shown to precipitate 50 to 100% serum thyrotropin, of which true macrotrothropin makes 56-98%. In the patients with subclinical hypothyroidism with a thyroid-stimulating hormone level of more than 10 pIU/ml, a trend towards an increase in the level of macrothyrotrophinaemia has been shown. The content of macrotyrotropin complex in patients with subclinical hypothyroidism, in whom the level of antibodies to thyroperoxidase is > 500 U/L, is significantly higher if compared to the patients with manifesting hypothyroidism. Elevated levels of antibodies to thyroperoxidase can lead to the generation of macrotyropin. Our findings have shown that the phenomenon of macrothyrotropinemia is quite common in patients with subclinical and manifesting hypothyroidism with Hashimoto thyroiditis (53.3%) and in control group (25%). Macrotyrotropin complex probably consists of thyrotropin and IgG. Patients with a thyroid-stimulating hormone level of > 10 pIU/ml are candidates for screening for the presence of the macrotyrotropin complex.The activity of the autoimmune process may correlate with the phenomenon of macrothyrotropinemia. The results can be used to develop an additional tool when choosing therapy in clinical practice

Боль в спине и оценка активности анкилозирующего спондилита на фоне гестации: симптомы, отражающие обострение заболевания, и беременность

Objective: to study the dynamics of back pain severity and the frequency of its individual components, characterizing the inflammatory and mechanical rhythm, on the background of gestation in women with ankylosing spondylitis (AS) and compare them with manifestations of dorsopathy in healthy pregnant women; to determine the information value of BASDAI during pregnancy, i.e. to identify its components with the optimal sensitivity and specificity ratio for assessing AS activity during gestation.Patients and methods. The main group consisted of 49 pregnant women with AS who met the modified New York criteria of 1984. The average age of patients was 31.7±4.9 years, the average duration of the disease was 134.4±85.8 months. BASDAI for trimesters of pregnancy was: 2.3 [1.2; 4.4], 2.8 [1.4; 4.2] and 2.2 [1.6; 4.0]. The control group included 51 pregnant women with back pain associated with gestation, without rheumatic diseases. The average age was 28.0±4.4 years. The intensity of back pain was assessed on a numerical rating scale. ROC-analysis was performed with calculation of the area under the curve (AUC) of each component of BASDAI in each pregnancy trimester.Results and discussion. More than 80% of pregnant women with AS experienced back pain, while the intensity of general back pain did not differ from that in the control group. Night back pain with improvement on awakening in the main group was present in 70%, 58% and 68.8% of women, respectively, in the I-III trimesters; its intensity was 3 [1; 5], 3.5 [3; 6] and 3.4 [2; 5] respectively; the frequency and severity of night pain were higher than in the control group. The number of women with improvement after exercise in the II and III trimesters did not differ in both groups. In the second half of pregnancy, 40% of AS patients noted improvement during rest, 52.1% – increased pain after exercise; the frequency of mechanical rhythm pain elements remained lower than in the control group. The AUC value for BASDAI in the first trimester was 0.74; AUC of all BASDAI components was >0.5. In the II and III trimesters, the AUC values for fatigue and back pain were 0.8.Conclusion. During pregnancy the vast majority of women with AS experience back pain, the nature of which changes in the second half of gestation. Night pain that improves on awakening reflects AS activity and is not related to pregnancy. The BASDAI components of severity and duration of morning stiffness have the highest classification value during pregnancy. Цель исследования – изучить динамику выраженности боли в спине и частоту ее отдельных компонентов, характеризующих воспалительный и механический ритм, на фоне гестации у женщин с анкилозирующим спондилитом (АС) и сравнить их с проявлениями дорсопатии у здоровых беременных; определить информативность BASDAI во время беременности, т. е. выявить его компоненты с оптимальным соотношением чувствительности и специфичности для оценки активности АС при гестации.Пациенты и методы. Основную группу составили 49 беременных с АС, соответствовавших модифицированным Нью-Йоркским критериям 1984 г. Средний возраст пациенток – 31,7±4,9 года, средняя продолжительность болезни – 134,4±85,8 мес. BASDAI по триместрам беременности равнялся: 2,3 [1,2; 4,4], 2,8 [1,4; 4,2] и 2,2 [1,6; 4,0]. В контрольную группу вошла 51 беременная с болью в спине, связанной с гестацией, без ревматических заболеваний. Средний возраст – 28,0±4,4 года. Интенсивность боли в спине оценивали по числовой рейтинговой шкале. Выполнен ROC-анализ с вычислением площади под кривой (AUC) каждого компонента BASDAI в каждом триместре беременности.Результаты и обсуждение. Более 80% беременных с АС испытывали боль в спине, при этом интенсивность общей боли в спине не отличалась от таковой в контрольной группе. Ночная боль в спине с улучшением при пробуждении в основной группе имелась у 70, 58 и 68,8% женщин соответственно в I–III триместрах; ее интенсивность составила 3 [1; 5], 3,5 [3; 6] и 3,4 [2; 5] соответственно; частота и выраженность ночной боли были выше, чем в контрольной группе. Количество женщин с улучшением после выполнения физических упражнений во II и III триместрах не различалось в обеих группах. Во второй половине беременности 40% больных АС отметили улучшение во время отдыха, 52,1% – усиление боли после физических упражнений; частота элементов боли механического ритма оставалась ниже, чем в контрольной группе. Значение AUC для BASDAI в I триместре составляло 0,74; AUC всех компонентов BASDAI была >0,5. Во II и III триместрах значения AUC для утомляемости и боли в спине оказались 0,8.Заключение. Подавляющее большинство женщин с АС во время беременности испытывают боль в спине, характер которой меняется во второй половине гестации. Ночная боль с улучшением при пробуждении отражает активность АС и не связана с беременностью. Наиболее высокой классификационной ценностью при беременности обладают такие компоненты BASDAI, как выраженность и длительность утренней скованности

Изменения гемоциркуляторных и респираторных функций при бронхиальной астме у детей

One hundred and fifty bronchial asthma patients aged 6 to 14 yrs were complexly examined for haemodynamic parameters in correlation with the lung function and the diaphragm activity in different stages of the disease. The respiratory dysfunction based on the bronchial obstruction, irregular lung ventilation and reduction in the diaphragm functional activity was shown to be accompanied by changes in the regional and central blood circulation. A consistency of these disorders depended on the basic therapyУ 150 больных бронхиальной астмой в возрасте от 6 до 14 лет проведено комплексное исследование параметров гемодинамики во взаимосвязи с оценкой функции внешнего дыхания и активности диафрагмы в различные периоды заболевания. Показано, что респираторная дисфункция, в основе которой лежат бронхиальная обструкция, неравномерная вентиляция легких и снижение функциональной активности диафрагмы, сочетается с изменениями регионарной и центральной гемоциркуляции. Причем стойкость отмеченных отклонений зависит от применения базисной терапии