<em>Galega officinalis</em> L. and Immunological Status in Diabetes Mellitus

Under diabetes mellitus, the administration of Galega officinalis promotes restoration of leukocyte precursors’ bone marrow pool and normalizes their proliferative activity. This plant protects the functional state of leukocytes by modulating actin cytoskeleton formation and through quantitative redistribution of leukocyte membrane glycoconjugates. Galega officinalis prevents the development of diabetes-associated oxidative stress which results in antiapoptotic activity. The normalization of leukocytes’ proliferative and functional capacity by Galega officinalis, along with its antiapoptotic and hypoglycemic effects, can improve the course of the disease and may prevent the development of complications of diabetes

Functional activities of neutrophils in diabetic rats are changed by yacon extracts

Introduction. Characterization of the polymorphonuclear neutrophils functional activity during the diabetes is of outmost importance for the understanding of the immunological processes involved in disease pathogenesis. The search for effective drugs that would suppress and/or modulate the immune response, preferably without negative effects, is a promising area of modern research. Smallanthus sonchifolius Poepp. et Endl., which has long been used in folk medicine to treat diabetes, was proposed as a potential treatment for immune disorders. The study aimed to search for the effective drug which would be able to regulate neutrophils functional activity in experimentally induced type 1 diabetes mellitus. Material and methods. The experiments were conducted on male Wistar rats. Diabetes was induced by intraabdominal injection of streptozotocin. For the evaluation of phagocytic activity and the content of cationic proteins and myeloperoxidase in neutrophils cytochemical studies were performed. Results. Diabetes development was accompanied by reduced phagocytic activity of granulocytes and increased activity of myeloperoxidase. All used extracts intensified phagocytic activity of neutrophil in diabetes, with the yacon root tubers extract demonstrating the strongest effect, which was reflected by rapid degradation of particles engulfed by phagocytes. Conclusions. It was established that yacon extracts inhibit myeloperoxidase activity in diabetes. Therefore all investigated extracts have a pronounced immunocorrective effect and can become the basis for creating a new generation of antidiabetic drugs

The Structure of Leukocyte Sialic Acid-Containing Membrane Glycoconjugates is a Differential Indicator of the Development of Diabetic Complications

Glycans, as potential prognostic biomarkers, deserve attention in clinical glycomics for diseases diagnosis. The variety of glycan chains, attached to proteins and lipids, makes it possible to form unique glycoconjugates with a wide range of cellular functions. Under leukocyte-endothelial interaction, not only the availability of glycoconjugates with sialic acids at the terminal position of glycans are informative, but also the type of glycosidic bond by which sialic acids links to subterminal carbohydrates in structure of glycans. The process of sialylation of leukocyte glycoconjugates undergoes considerable changes in type 1 diabetes mellitus. At early stage of disease without diabetic complications, the pathology is accompanied by the increase of α2,6-linked sialic acids. The quantity of sialic acid-containing glycoconjugates on leukocytes surface increases in condition of disease duration up to five years. However, the quantity of sialic acids linked by α2,6-glycosidic bonds decreases in patients with the disease duration over ten years. Therefore, sialoglycans as marker molecules determine the leukocyte function in patients with type 1 diabetes mellitus, depending on the disease duration. Changes in the glycans structure of membrane glycoconjugates of leukocytes allow understanding the mechanism of diabetic complications development

Red Wine and Yacon as a Source of Bioactive Compounds with Antidiabetic and Antioxidant Potential

Phytochemicals derived from different plants are promising therapeutic agents. Herbal compounds can be used under diseases, etiological causes of which are alterations of carbohydrate, protein, and lipid metabolisms, along with increased oxidative stress and chronic low-grade inflammation. Potential sources of biologically active substances may be grape wine, rich in phenolic compounds. Well-studied examples of polyphenols are phenolic acids, catechins, anthocyanins, and flavonoids, etc. Another source of biologically active compounds is yacon (Smallanthus sonchifolius Poepp. & Endl.). The aboveground part of yacon is rich in phenolic compounds and terpenes. Main biologically active substances from tuberous roots of yacon are fructooligosaccharides and phenolic compounds. The section will be devoted to the analysis of hypoglycemic and antioxidant effects, and molecular targets of the complex of biologically active substances derived from red wine and yacon

The dynamics of actin filament polymerization in activated leukocytes under experimental diabetes mellitus against the background of agmatine administration

Aim. To research the impact of agmatine on the redistribution of actin fractions, which are repre-sented by cytoskeletal actin filaments, short actin filaments of the plasma membrane skeleton and actin monomers (G-actin) in rat leukocytes under experimental diabetes mellitus (EDM). Methods. Leukocytes were lyzed in Triton X-100 and subjected to centrifugation to obtain cytoskeletal actin filaments, short actin filaments and actin monomers, which were separated in SDS-PAAG, followed by the immunoblot analysis of anti-actin antibodies. Results. Under EDM, an intensifi-cation of the process of short actin filament depolymerization and an increase in the G-actin con-tent were observed in leukocytes activated by sialospecific wheat germ lectin (WGA). Against a background of agmatine administration, the response dynamics to the WGA-stimulating effect was characterized by an increase in actin polymerization in the fraction of cytoskeletal filaments already after 0.5 min exposure to lectin, an exceptionally rapid depolymerization process after 1 min of the lectin treatment, and return to the initial indices after 3 min exposure to lectin. Conclusions. In leukocytes of animals with EDM against a background of agmatine administration, the transduction of WGA-induced signal through sialoglycoconjugates causes the actin redistribution. It indicates that this polyamine helps to restore and maintain a functional response of leukocytes to the activation signals.Мета. Дослідити вплив агматину на перерозподіл фракцій актину, які представлені актиновими філаментами цито-скелету, короткими актиновими філаментами скелету плазматичної мембрани і мономерами актину (G-актин), у лейкоцитах щурів з експериментальним цукровим діабетом (ЕЦД) Методи. Лейкоцити лізували в Тритоні Х-100 і піддавали центрифугуванню, у результаті якого було отримано актинові філаменти цитоскелету, короткі актинові філаменти і мономери актину, які розділяли в SDS-ПААГ, після чого проводили імуноблот аналіз із використан-ням анти-актинових антитіл. Результати. За умов ЕЦД у лейкоцитах, активованих сіалоспецифічним лектином зародків пшениці (WGA), спостерігається інтенсифікування процесу деполімеризації коротких актинових філаме-нтів і збільшення вмісту G-актину, а на фоні введення агматину динаміка формування відповіді на WGA-стимулювальний вплив характеризувалася посиленням полімеризації актину у фракції філаментів цитоскелету вже на 0,5 хв впливу лектину, дуже стрімким процесом деполімеризації на 1 хв після дії лектину та поверненням пока-зників до рівня у вихідному стані у разі дії лектину впродовж 3 хв. Висновки. У лейкоцитах тварин з ЕЦД на фоні введення агматину трансдукція WGA-індукованого сигналу через сіалоглікокон’югати зумовлює перерозподіл вмісту актину, вказуючи на те, що даний поліамін сприяє відновленню і підтриманню функціональної відповіді лейкоцитів на активаційні сигнали.Цель. Исследовать влияние агматина на перераспределение фракций актина, которые представлены актиновыми филаментами цитоскелета, короткими актиновыми филаментами скелета плазматической мембраны и мономера-ми актина (G-актин), в лейкоцитах крыс с экспериментальным сахарным диабетом (ЭСД). Методы. Лейкоциты лизировали в Тритоне Х-100 и подвергали центрифугированию, в результате которого было получено актиновые филаменты цитоскелета, короткие актиновые филаменты и мономеры актина, которые разделяли в SDS-ПААГ, после чего проводили иммуноблот анализ с использованием антиактиновых антител. Результаты. В условиях ЭСД в лейкоцитах, активированных сиалоспецыфическим лектином зародышей пшеницы (WGA), наблюдается интен-сификация процесса деполимеризации коротких актиновых филаментов и увеличение содержания G-актина, а на фоне введения агматина динамика формирования ответа на WGA-стимулирующее влияние характеризировалась усилением полимеризации актина во фракции филаментов цитоскелета уже на 0,5 мин влияния лектина, очень стремительным процессом деполимеризации на 1 мин после действия лектина и возвращением показателей к уро-вню в исходном состоянии в случае действия лектина в течение 3 мин. Выводы. В лейкоцитах животных с ЭСД на фоне введения агматина трансдукция WGA-индуцированого сигнала через сиалогликоконьюгаты приводит к пе-рераспределению содержания актина, указывая на то, что данный полиамин способствует восстановлению и по-ддержанию функционального ответа лейкоцитов на активационные сигналы

Shewanella oneidensis: a new and efficient System for Expression and Maturation of heterologous [Fe-Fe] Hydrogenase from Chlamydomonas reinhardtii

<p>Abstract</p> <p>Background</p> <p>The eukaryotic green alga, <it>Chlamydomonas reinhardtii</it>, produces H₂ under anaerobic conditions, in a reaction catalysed by a [Fe-Fe] hydrogenase HydA1. For further biochemical and biophysical studies a suitable expression system of this enzyme should be found to overcome its weak expression in the host organism. Two heterologous expression systems used up to now have several advantages. However they are not free from some drawbacks. In this work we use bacterium <it>Shewanella oneidensis</it> as a new and efficient system for expression and maturation of HydA1 from <it>Chlamydomonas reinhardtii</it>.</p> <p>Results</p> <p>Based on codon usage bias and hydrogenase maturation ability, the bacterium <it>S. oneidensis</it>, which possesses putative [Fe-Fe] and [Ni-Fe] hydrogenase operons, was selected as the best potential host for <it>C. reinhardtii </it>[Fe-Fe] hydrogenase expression. Hydrogen formation by <it>S. oneidensis </it>strain AS52 (Δ<it>hydA</it>Δ<it>hyaB</it>) transformed with a plasmid bearing <it>Cr</it>HydA1 and grown in the presence of six different substrates for anaerobic respiration was determined. A significant increase in hydrogen evolution was observed for cells grown in the presence of trimethylamine oxide, dimethylsulfoxide and disodium thiosulfate, showing that the system of <it>S. oneidensis </it>is efficient for heterologous expression of algal [Fe-Fe] hydrogenase.</p> <p>Conclusion</p> <p>In the present work a new efficient system for heterologous expression and maturation of <it>C. reinhardtii </it>hydrogenase has been developed. HydA1 of <it>C. reinhardtii </it>was purified and shown to contain 6 Fe atoms/molecule of protein, as expected. Using DMSO, TMAO or thiosulfate as substrates for anaerobic respiration during the cell growth, 0.4 – 0.5 mg l^-1(OD₆₀₀= 1) of catalytically active HydA1 was obtained with hydrogen evolution rate of ~700 μmol H₂mg^-1min^-1.</p

The structure and function of mitochondria-associated endoplasmic reticulum membranes and their role in pancreatic β-cells dysregulation

Membrane trafficking and organelle contact sites are important for regulating cell metabolism and survival. The highly specialized regions of close contacts between mitochondria and endoplasmic reticulum (ER), called mitochondria associated membranes (MAMs), are crucial signaling hubs for the lipid and calcium homeostasis, reactive oxygen species delivery, regulation of autophagy and mitochondrial dynamics. In recent years, MAMs have been the focus of multiple studies for identifying the MAMs proteins and defining their signaling mechanisms. Many studies have proved the importance of MAMs in maintaining the normal function of both organelles. Excessive MAM formation is known to trigger the cascade of pathological events, such as mitochondria calcium overload, aberrant lipid levels, autophagosome formation, and eventually, cell apoptosis. In this article, we focus on the composition and function of MAMs, more specifically, the role of MAMs in Ca2+ uptake, ER stress, mitochondrial fusion and fission and autophagy. The altered interaction between ER and mitochondria results in the amendment of pancreatic tissues, revealing the role of MAMs in glucose homeostasis and the development of diabetes. The development of mitochondrial dysfunction, ER stress and oxidative stress are co-related with β-cell dysfunction. MAMs are likely to play an important role of the functional state regulation in pancreatic cells under pathologies by regulating the signaling of the two organelles and the crosstalk of the two pathological events. It was found that under streptozotocin-induced diabetes, the increased level of mitophagy in pancreatic tissue is connected with tight junctions of MAMs

Diabetes-correcting and antioxidant effects of grape pomace extract rich in natural complex of polyphenols

Background. The positive health effects of polyphenols have led to an increased scientific interest in these natural compounds over the past decade. Many studies confirm the effectiveness of polyphenols as additional therapy in diabetes, especially due to the sugar-lowering effect of polyphenols. The aim of the research was to investigate the morphological and functional state of peripheral blood erythrocytes and the indices of oxidative stress in the liver of rats with experimental diabetes and after the administration of grape pomace extract rich in natural complex of polyphenols. Materials and Methods. We obtained grape pomace extract, which contains a variety of polyphenolic compounds. Rats of the following groups were used in the experiments: control animals, animals treated with grape pomace extract rich in natural complex of polyphenols for 14 days, animals with streptozotocin-induced diabetes mellitus, and animals with streptozotocin-induced diabetes mellitus treated with grape pomace extract rich in natural complex of polyphenols for 14 days. The number of erythrocytes and reticulocytes, the concentration of hemoglobin and glycated hemoglobin were determined in the peripheral blood of rats. The activities of catalase, superoxide dismutase and the content of thiobarbituric acid-reactive substance and carbonyl groups of proteins were determined in the liver tissues of rats. Results. The study has shown an increase in the number of erythrocytes and the level of hemoglobin, a decrease in the level of glycated hemoglobin and the number of reticulocytes in the peripheral blood of rats after administration of grape pomace extract rich in natural complex of polyphenols to rats with experimental diabetes. A decrease in the content of thiobarbituric acid-reactive substance and the content of carbonyl groups of proteins of neutral and basic character and an increase in the activity of catalase and superoxide dismutase in liver tissues were found under the same conditions. Conclusions. The results indicate that the extract of the natural complex of polyphenols is capable of correcting the morphological and functional state of erythrocytes, as well as improving the activity of antioxidant enzymes and the content of marker molecules of oxidative stress in hepatocytes of rats under experimental diabetes mellitus

Testing the role of SOX15 in human primordial germ cell fate.

Background: Potentially novel regulators of early human germline development have been identified recently, including SOX15 and SOX17, both of which show specific expression in human primordial germ cells. SOX17 is now known to be a critical specifier of human germ cell identity. There have been suggestions, as yet without evidence, that SOX15 might also play a prominent role. The early human germline is inaccessible for direct study, but an in vitro model of human primordial germ cell-like cell (hPGCLC) specification from human embryonic stem cells (hESCs) has been developed. This enables mechanistic study of human germ cell specification using genetic tools to manipulate the levels of SOX15 and SOX17 proteins to explore their roles in hPGCLC specification. Methods: SOX15 and SOX17 proteins were depleted during hPGCLC specification from hESCs using the auxin-inducible degron system, combined with a fluorescent reporter for tracking protein levels. Additionally, SOX15 protein was overexpressed using the ProteoTuner system. Protein-level expression changes were confirmed by immunofluorescence. The impact on hPGCLC specification efficiency was determined by flow cytometry at various time points. qPCR experiments were performed to determine some transcriptional effects of SOX15 perturbations. Results: We observed specific SOX15 expression in hPGCLCs by using immunofluorescence and flow cytometry analysis. Depletion of SOX15 had no significant effect on hPGCLC specification efficiency on day 4 after induction, but there was a significant and progressive decrease in hPGCLCs on days 6 and 8. By contrast, depletion of SOX17 completely abrogated hPGCLC specification. Furthermore, SOX15 overexpression resulted in a significant increase in hPGCLC fraction on day 8. qPCR analysis revealed a possible role for the germ cell and pluripotency regulator PRDM14 in compensating for changes to SOX15 protein levels. Conclusions: SOX17 is essential for hPGCLC specification, yet SOX15 is dispensable. However, SOX15 may have a role in maintaining germ cell identity