EGCG as an anti-SARS-CoV-2 agent: Preventive versus therapeutic potential against original and mutant virus
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01.12.2021 |
Tsvetkov V.
Varizhuk A.
Kozlovskaya L.
Shtro A.
Lebedeva O.
Komissarov A.
Vedekhina T.
Manuvera V.
Zubkova O.
Eremeev A.
Shustova E.
Pozmogova G.
Lioznov D.
Ismukhametov A.
Lazarev V.
Lagarkova M.
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Biochimie |
10.1016/j.biochi.2021.08.003 |
0 |
Ссылка
In the search for anti-SARS-CoV-2 drugs, much attention is given to safe and widely available native compounds. The green tea component epigallocatechin 3 gallate (EGCG) is particularly promising because it reportedly inhibits viral replication and viral entry in vitro. However, conclusive evidence for its predominant activity is needed. We tested EGCG effects on the native virus isolated from COVID-19 patients in two independent series of experiments using VERO cells and two different treatment schemes in each series. The results confirmed modest cytotoxicity of EGCG and its substantial antiviral activity. The preincubation scheme aimed at infection prevention has proven particularly beneficial. We complemented that finding with a detailed investigation of EGCG interactions with viral S-protein subunits, including S2, RBD, and the RBD mutant harboring the N501Y mutation. Molecular modeling experiments revealed N501Y-specific stacking interactions in the RBD-ACE2 complex and provided insight into EGCG interference with the complex formation. Together, these findings provide a molecular basis for the observed EGCG effects and reinforce its prospects in COVID-19 prevention therapy.
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Alterations in Blood Metabolic Parameters of Immature Mice After Subchronic Exposure to Cobalt Chloride
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01.02.2021 |
Vladov I.
Petrova E.
Pavlova E.
Tinkov A.A.
Ajsuvakova O.P.
Skalny A.V.
Gluhcheva Y.
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Biological Trace Element Research |
10.1007/s12011-020-02161-4 |
0 |
Ссылка
© 2020, Springer Science+Business Media, LLC, part of Springer Nature. The wide use of cobalt (Co) in food, industry, and medical devices requires full elucidation of its biological effects on tissues and organs. The aim was to assess serum metabolic alterations in immature mice after subchronic exposure to CoCl2. Pregnant ICR mice were subjected to a daily dose of 75 mg cobalt chloride/kg body weight (CoCl2x6H2O) 2–3 days before they gave birth, and treatment continued until days 25 and 30 after delivery. The compound was dissolved in and obtained with regular tap water. ICP-DRC-MS analysis showed significantly elevated serum Co2+ and diverse alterations in metabolic parameters of 25- and 30-day-old pups after exposure to CoCl2. Cholesterol and urea levels were significantly elevated in day 25 mice while HDL-C and LDL-C were reduced. In day 30, Co-exposed mice LDL-C and triglycerides were significantly increased while the total cholesterol level remained unchanged. Alkaline phosphatase was significantly reduced in day 25 Co-exposed mice. Blood glucose level of Co-exposed mice remained close to the untreated controls. Total protein content was slightly increased in day 30 mice. Co-exposure reduced albumin content and albumin/globulin ratio but increased significantly globulin content. Co administration showed strong correlation with cholesterol, urea, and HDL-C in both day 25 and 30 mice. Inverse correlation was found with alkaline phosphatase and albumin for day 25 and with triglycerides, globulin, and total protein content in day 30 Co-exposed mice. Subchronic CoCl2 exposure of immature mice induced significant changes in key metabolic parameters suggesting possible further disturbances in energy metabolism, osteogenesis, and reproduction.
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Alterations in Blood Metabolic Parameters of Immature Mice After Subchronic Exposure to Cobalt Chloride
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01.02.2021 |
Vladov I.
Petrova E.
Pavlova E.
Tinkov A.A.
Ajsuvakova O.P.
Skalny A.V.
Gluhcheva Y.
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Biological Trace Element Research |
10.1007/s12011-020-02161-4 |
0 |
Ссылка
© 2020, Springer Science+Business Media, LLC, part of Springer Nature. The wide use of cobalt (Co) in food, industry, and medical devices requires full elucidation of its biological effects on tissues and organs. The aim was to assess serum metabolic alterations in immature mice after subchronic exposure to CoCl2. Pregnant ICR mice were subjected to a daily dose of 75 mg cobalt chloride/kg body weight (CoCl2x6H2O) 2–3 days before they gave birth, and treatment continued until days 25 and 30 after delivery. The compound was dissolved in and obtained with regular tap water. ICP-DRC-MS analysis showed significantly elevated serum Co2+ and diverse alterations in metabolic parameters of 25- and 30-day-old pups after exposure to CoCl2. Cholesterol and urea levels were significantly elevated in day 25 mice while HDL-C and LDL-C were reduced. In day 30, Co-exposed mice LDL-C and triglycerides were significantly increased while the total cholesterol level remained unchanged. Alkaline phosphatase was significantly reduced in day 25 Co-exposed mice. Blood glucose level of Co-exposed mice remained close to the untreated controls. Total protein content was slightly increased in day 30 mice. Co-exposure reduced albumin content and albumin/globulin ratio but increased significantly globulin content. Co administration showed strong correlation with cholesterol, urea, and HDL-C in both day 25 and 30 mice. Inverse correlation was found with alkaline phosphatase and albumin for day 25 and with triglycerides, globulin, and total protein content in day 30 Co-exposed mice. Subchronic CoCl2 exposure of immature mice induced significant changes in key metabolic parameters suggesting possible further disturbances in energy metabolism, osteogenesis, and reproduction.
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Novel applications of modification of thiol enzymes and redox-regulated proteins using S-methyl methanethiosulfonate (MMTS)
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01.11.2019 |
Makarov V.
Tikhomirova N.
Savvateeva L.
Petushkova A.
Serebryakova M.
Baksheeva V.
Gorokhovets N.
Zernii E.
Zamyatnin A.
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Biochimica et Biophysica Acta - Proteins and Proteomics |
10.1016/j.bbapap.2019.07.012 |
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© 2019 Elsevier B.V. S-Methyl methanethiosulfonate (MMTS) is used in experimental biochemistry for alkylating thiol groups of protein cysteines. Its applications include mainly trapping of natural thiol-disulfide states of redox-sensitive proteins and proteins which have undergone S-nitrosylation. The reagent can also be employed as an inhibitor of enzymatic activity, since nucleophilic cysteine thiolates are commonly present at active sites of various enzymes. The advantage of using MMTS for this purpose is the reversibility of the formation of methylthio mixed disulfides, compared to irreversible alkylation using conventional agents. Additional benefits include good accessibility of MMTS to buried protein cysteines due to its small size and the simplicity of the protection and deprotection procedures. In this study we report examples of MMTS application in experiments involving oxidoreductase (glyceraldehyde-3-phosphate dehydrogenase, GAPDH), redox-regulated protein (recoverin) and cysteine protease (triticain-α). We demonstrate that on the one hand MMTS can modify functional cysteines in the thiol enzyme GAPDH, thereby preventing thiol oxidation and reversibly inhibiting the enzyme, while on the other hand it can protect the redox-sensitive thiol group of recoverin from oxidation and such modification produces no impact on the activity of the protein. Furthermore, using the example of the papain-like enzyme triticain-α, we report a novel application of MMTS as a protector of the primary structure of active cysteine protease during long-term purification and refolding procedures. Based on the data, we propose new lines of MMTS employment in research, pharmaceuticals and biotechnology for reversible switching off of undesirable activity and antioxidant protection of proteins with functional thiol groups.
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Proteome Integral Solubility Alteration: A High-Throughput Proteomics Assay for Target Deconvolution
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01.11.2019 |
Gaetani M.
Sabatier P.
Saei A.
Beusch C.
Yang Z.
Lundström S.
Zubarev R.
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Journal of Proteome Research |
10.1021/acs.jproteome.9b00500 |
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© 2019 American Chemical Society. Various agents, including drugs as well as nonmolecular stimuli, induce alterations in the physicochemical properties of proteins in cell lysates, living cells, and organisms. These alterations can be probed by applying a stability- and solubility-modifying factor, such as elevated temperature, to a varying degree. As a second dimension of variation, drug concentration or agent intensity/concentration can be used. Compared to standard approaches where curves are fitted to protein solubility data acquired at different temperatures and drug concentrations, Proteome Integral Solubility Alteration (PISA) assay increases the analysis throughput by 1 to 2 orders of magnitude for an unlimited number of factor variation points in such a scheme. The consumption of the compound and biological material decreases in PISA by the same factor. We envision widespread use of the PISA approach in chemical biology and drug development.
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Structural Transition States Explored With Minimalist Coarse Grained Models: Applications to Calmodulin
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15.10.2019 |
Delfino F.
Porozov Y.
Stepanov E.
Tamazian G.
Tozzini V.
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Frontiers in Molecular Biosciences |
10.3389/fmolb.2019.00104 |
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© Copyright © 2019 Delfino, Porozov, Stepanov, Tamazian and Tozzini. Transitions between different conformational states are ubiquitous in proteins, being involved in signaling, catalysis, and other fundamental activities in cells. However, modeling those processes is extremely difficult, due to the need of efficiently exploring a vast conformational space in order to seek for the actual transition path for systems whose complexity is already high in the stable states. Here we report a strategy that simplifies this task attacking the complexity on several sides. We first apply a minimalist coarse-grained model to Calmodulin, based on an empirical force field with a partial structural bias, to explore the transition paths between the apo-closed state and the Ca-bound open state of the protein. We then select representative structures along the trajectory based on a structural clustering algorithm and build a cleaned-up trajectory with them. We finally compare this trajectory with that produced by the online tool MinActionPath, by minimizing the action integral using a harmonic network model, and with that obtained by the PROMPT morphing method, based on an optimal mass transportation-type approach including physical constraints. The comparison is performed both on the structural and energetic level, using the coarse-grained and the atomistic force fields upon reconstruction. Our analysis indicates that this method returns trajectories capable of exploring intermediate states with physical meaning, retaining a very low computational cost, which can allow systematic and extensive exploration of the multi-stable proteins transition pathways.
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Role of heterotrimeric G proteins in platelet activation and clot formation in platelets treated with integrin αIIbβ3 inhibitor
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03.04.2018 |
Budnik I.
Shenkman B.
Hauschner H.
Zilinsky I.
Savion N.
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Platelets |
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2 |
Ссылка
© 2018 Taylor & Francis. Mechanisms of platelet activation are triggered by thrombin, adenosine diphosphate (ADP), epinephrine, thromboxane A2, and other soluble agonists which induce signaling via heterotrimeric Gαq, Gαi, and Gα12/13 proteins. We have undertaken a study addressing the contribution of these G proteins to platelet activation and clot formation in the presence of eptifibatide, thus excluding outside-in signaling provided by integrin αIIbβ3–fibrinogen engagement. Selective and combined activation of the G proteins was achieved by using combinations of platelet agonists and inhibitors. Platelet activation in platelet-rich plasma was evaluated by P-selectin expression using flow cytometry. Contribution of platelets to whole blood clotting was assessed by rotation thromboelastometry (ROTEM). Selective signaling of Gαq or Gαi but not Gα12/13 promoted P-selectin expression. Further enhancement of P-selectin expression was achieved by ADP-induced combined signaling of Gαq and Gαi, and to more extent by U46619 at high concentration (1.5 μM) induced combined signaling of Gαq and Gα12/13 while maximal P-selectin expression was achieved by thrombin receptor-activating peptide (TRAP)-induced combined signaling of Gαq, Gαi, and Gα12/13. In ROTEM, selective activation of Gαq, Gαi, or Gα12/13 failed to affect blood clotting. Combined signaling of Gαq and Gαi or Gαq and Gα12/13 or all three G proteins shortened the clotting time and stimulated clot strength. Pretreatment of platelets with acetylsalicylic acid did not change the effect of ADP but inhibited the effect of TRAP. Signaling of Gαq and Gα12/13 triggered by U46619 also stimulated clot formation. Combined signaling of either Gαq and Gαi or Gαq and Gα12/13 is sufficient to stimulate maximal platelet activation and enhanced clot formation in platelets treated with inhibitor of integrin αIIbβ3. It could be suggested that outside-in signaling is not necessarily required to fulfill these platelet functions.
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Effects of laser radiation on mitochondria and mitochondrial proteins subjected to nitric oxide
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01.04.2018 |
Osipov A.
Machneva T.
Buravlev E.
Vladimirov Y.
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Frontiers in Medicine |
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0 |
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© 2018 Osipov, Machneva, Buravlev and Vladimirov. The biological roles of heme and nonheme nitrosyl complexes in physiological and pathophysiological conditions as metabolic key players are considered in this study. Two main physiological functions of protein nitrosyl complexes are discussed-(1) a depot and potential source of free nitric oxide (NO) and (2) a controller of crucial metabolic processes. The first function is realized through the photolysis of nitrosyl complexes (of hemoglobin, cytochrome c, or mitochondrial iron-sulfur proteins). This reaction produces free NO and subsequent events are due to the NO physiological functions. The second function is implemented by the possibility of NO to bind heme and nonheme proteins and produce corresponding nitrosyl complexes. Enzyme nitrosyl complex formation usually results in the inhibition (or enhancement in the case of guanylate cyclase) of its enzymatic activity. Photolysis of protein nitrosyl complexes, in this case, will restore the original enzymatic activity. Thus, cytochrome c acquires peroxidase activity in the presence of anionic phospholipids, and this phenomenon can be assumed as a key step in the programmed cell death. Addition of NO induces the formation of cytochrome c nitrosyl complexes, inhibits its peroxidase activity, and hinders apoptotic reactions. In this case, photolysis of cytochrome c nitrosyl complexes will reactivate cytochrome c peroxidase activity and speed up apoptosis. Control of mitochondrial respiration by NO by formation or photolytic decay of iron-sulfur protein nitrosyl complexes is an effective instrument to modulate mitochondrial metabolism. These questions are under discussion in this study.
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Effects of laser radiation on mitochondria and mitochondrial proteins subjected to nitric oxide
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01.04.2018 |
Osipov A.
Machneva T.
Buravlev E.
Vladimirov Y.
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Frontiers in Medicine |
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1 |
Ссылка
©2018 Osipov, Machneva, Buravlev and Vladimirov. The biological roles of heme and nonheme nitrosyl complexes in physiological and pathophysiological conditions as metabolic key players are considered in this study. Two main physiological functions of protein nitrosyl complexes are discussed-(1) a depot and potential source of free nitric oxide (NO) and (2) a controller of crucial metabolic processes. The first function is realized through the photolysis of nitrosyl complexes (of hemoglobin, cytochrome c, or mitochondrial iron-sulfur proteins). This reaction produces free NO and subsequent events are due to the NO physiological functions. The second function is implemented by the possibility of NO to bind heme and nonheme proteins and produce corresponding nitrosyl complexes. Enzyme nitrosyl complex formation usually results in the inhibition (or enhancement in the case of guanylate cyclase) of its enzymatic activity. Photolysis of protein nitrosyl complexes, in this case, will restore the original enzymatic activity. Thus, cytochrome c acquires peroxidase activity in the presence of anionic phospholipids, and this phenomenon can be assumed as a key step in the programmed cell death. Addition of NO induces the formation of cytochrome c nitrosyl complexes, inhibits its peroxidase activity, and hinders apoptotic reactions. In this case, photolysis of cytochrome c nitrosyl complexes will reactivate cytochrome c peroxidase activity and speed up apoptosis. Control of mitochondrial respiration by NO by formation or photolytic decay of iron-sulfur protein nitrosyl complexes is an effective instrument to modulate mitochondrial metabolism. These questions are under discussion in this study.
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Cervical intraepithelial neoplasia associated with papillomavirus infection: Pathogenetic rationale of treating patients during the reproductive period
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01.01.2018 |
Davydov A.
Shakhlamova M.
Lebedev V.
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Voprosy Ginekologii, Akusherstva i Perinatologii |
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3 |
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© 2018 Dynasty Publishing House. All rights reserved. The lecture deals with treatment of patients with CIN associated with papillomavirus infection (HPVI). The issues of epidemiology, diagnosis and treatment of CIN are discussed. Emphasis is put on a complex approach to treating patients with HPVI-associated CIN. Special attention is paid to studying the mechanisms of dysregulation of immune response during the period of human papillomavirus virus (HPV) persistence, which promotes immune suppression and is considered to be a necessary precondition for progression of HPV-associated cancer. Based on analysis of literature sources, the authors show that high oncogenic risk E6 and E7 HPV types interact with key proteins of interferon signalling pathway, inhibiting its production. This accounts for insufficient effectiveness of preparations of pure interferons and their inductors for treatment of HPV infection. From the pathogenetic positions, the use of inosine pranobex as a medication in complex therapy would be appropriate, since it ensures complete elimination of HPV and reduces the frequency of HPV infection recurrences.
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Correction of anxiety disorders: Focus on a comorbid patient
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01.01.2018 |
Shavlovskaya O.
Kuznetsov S.
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Terapevticheskii Arkhiv |
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0 |
Ссылка
© 2018 Media Sphera Publishing Group. All rights reserved. The exact cause of the development of anxiety disorders (AD) in present time has not been fully established and is a subject of debate in many countries. Interest in studying the mechanisms of action of proteins of S100 group, in particular, neurospecific protein S100b, is caused by its participation in processes of integrative activity of brain/neuron and development of diseases of nervous system. The functions of S100 proteins determine their influence on synaptic plasticity and participation in the regulation of stress-realizing and stress-limiting systems, the imbalance of which (primarily, the insufficiency of the GABA-ergic system) is the neurobiological basis of the majority of anxiety-depressive pathologies. Preparations regulating the activity of S100 protein have a distinct clinical anti-anxiety effect and additionally contribute to the restoration of neuronal plasticity processes.
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Vibrational spectroscopy of tissue-engineered structures based on proteins, chitosan, and carbon nanotube conjugates
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01.01.2018 |
Polokhin A.
Fedorova Y.
Gerasimenko A.
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Proceedings of SPIE - The International Society for Optical Engineering |
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1 |
Ссылка
© 2018 SPIE. In this work, tissue-engineered structures based on a matrix of protein conjugates, chitosan and carbon nanotubes were prepared and studied. Bovine serum albumin (BSA), bovine collagen (BCrossed D sign¡) were used. Two types of single-walled carbon nanotubes (SWCNTs) were used to form a strong internal scaffold in a protein-chitosan matrix under the action of laser radiation. Tissue-engineered structures were created by means of layered deposition and laser evaporation of the initial aqueous dispersion from SWCNT, BSA, BC and chitosan succinate. As sources of laser radiation, a continuous diode laser with a wavelength of 810 nm and a pulsed fiber laser with a wavelength of 1064 nm and frequency of 80 kHz were used. Studies of tissue-engineered structures were carried out using vibrational spectroscopy methods (IR and Raman). The changes in the frequencies and intensities of the corresponding absorption bands and Raman lines of the amide group oscillations were analyzed. IR spectra of tissue-engineered structures demonstrated a high degree of binding of organic (protein, chitosan) and inorganic (SWCNT) components. The structure and defectiveness of the carbon nanotube scaffold were investigated in the Raman spectra.
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Effect of Chronic Alcohol Abuse on Anabolic and Catabolic Signaling Pathways in Human Skeletal Muscle
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01.01.2018 |
Shenkman B.
Belova S.
Zinovyeva O.
Samkhaeva N.
Mirzoev T.
Vilchinskaya N.
Altaeva E.
Turtikova O.
Kostrominova T.
Nemirovskaya T.
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Alcoholism: Clinical and Experimental Research |
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3 |
Ссылка
Copyright © 2017 by the Research Society on Alcoholism Background: Animal studies showed that alcoholic myopathy is characterized by the reduction in myofiber cross-sectional area (CSA) and by impaired anabolic signaling. The goal of this study was to compare changes in CSA and fiber type composition with modifications in anabolic and catabolic signaling pathways at the early stages of alcohol misuse in humans. Methods: Skeletal muscle samples from 7 male patients with chronic alcohol abuse (AL; 47.7 ± 2.0 years old; alcohol misuse duration 7.7 ± 0.6 years) were compared with muscle from a control group of 7 healthy men (C; 39.7 ± 5.0 years old). Biopsies from vastus lateralis muscles were taken and analyzed for the changes in fiber type composition, fiber CSA, and for the alterations in anabolic and catabolic signaling pathways. Results: AL patients did not have detectable clinical myopathy symptoms or muscle fiber atrophy, but the relative proportion of fast fibers was increased. There was a significant decrease in IGF-1 in plasma and IRS-1 protein content in muscle of AL group. Levels of total and phosphorylated p70S6K1, GSK3β, and p90RSK1 were not different between AL and C groups. Muscle of AL patients had increased mRNA expression of HSP70 and HSP90. A marker of anabolic pathway p-4E-BP1 was decreased, while catabolic markers (MuRF-1, MAFbx, ubiquitinated proteins) were increased in AL patients when compared with C group. Conclusions: At the early stages of alcohol misuse in humans, changes in the regulation of anabolic and catabolic signaling pathways precede the development of skeletal muscle atrophy and manifestation of clinical symptoms of alcoholic myopathy.
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The dopaminergic dysfunction and altered working memory performance of aging mice lacking gamma-synuclein gene
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01.01.2018 |
Kokhan V.
Kokhan T.
Samsonova A.
Fisenko V.
Ustyugov A.
Aliev G.
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CNS and Neurological Disorders - Drug Targets |
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0 |
Ссылка
© 2018 Bentham Science Publishers. Background: It was previously shown that inactivation of gamma-synuclein which is a small soluble neuronal protein affects psycho-emotional status and cognitive abilities in knock-out mice. Objective: Determine the role of gamma-synuclein inactivation on memory performance in aging animals. Method: We used the passive avoidance test and acute amphetamine administration in aging gammasynuclein knock-out mice. Results: As a result, we found moderate aging-unlinked deficit of dopaminergic neurotransmitter system of gamma-synuclein knock-out mice. At the same time, the evidence of progressive synaptic vesicle trafficking machinery impairment was obtained. Conclusion: Therefore most likely these dysfunctions are associated with a reduction in the highefficient learning performance in tests that require intact working memory.
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Molecular and cell mechanisms of the stress-protective activity of adaptogenic phytopreparation kardecaim on the background of emotional stress
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01.01.2018 |
Alekseeva E.
Malyshev I.
Shantanova L.
Nikolaev S.
Kareva E.
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Eksperimental'naya i Klinicheskaya Farmakologiya |
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0 |
Ссылка
© 2018 Izdatel'stvo Meditsina. All rights reserved. We have studied the adaptogenic properties of Kardecaim phytopreparation based on dry extracts derived from Inula helenium L., Zingiber officinale Roscoe, Elletaria cardamomum (L.) Maton., and Caragaha spinose (L.) Wall, ex Homem, which contain phenolic compounds and terpenoids. A course of preventive administration of Kardecaim to white rats in a dose of 100 mg/kg for 7 days before the induction of acute emotional stress produced stress-protective effect by preventing the development of catabolic signs of Selye's triad in animal inner organs, including the thymus involution (40% lower than in control); degree of hemorrhagic and ulcerative defects in the stomach mucosa (5 times less than reference gastroprotector effect), and somewhat less pronounced effect on the stress-induced hypertrophy of adrenal glands, yet comparable to the effect of reference preparation (eleutherococcus extract). It was shown that the increase in the resistance to stress under the action of Kardecaim was due to the activation of synthesis of heat shock proteins (Hsp). The Hsp-70 content in the thymus of white rats undergoing emotional stress was 56% higher than in the control. The introduction of Kardecaim to intact animals had no effect on the basal level of Hsp-70. It was established that the expression of Hsp under the drug action was not connected with NO system, since the administration of Kardecaim on the background of stress impact was followed by a decrease in the NO metabolite yield.
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Analysis of the expression levels of genes that encode cytoskeletal proteins in Drosophila melanogaster larvae during micro- and hypergravity effect simulations of different durations
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Огнева И. В.
Свистунов А.А
Несвижский Юрий Владимирович
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Biophysics (Russian Federation) |
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The goal of this study was to find genes that encode cytoskeletal proteins that are potential candidates for the role of triggers in cell mechanosensitivity in the fruit fly. Centrifugation was used to simulate the hypergravity effects (2g group); the constantly changing orientation of the larvae in the gravity field was performed in order to simulate the effects of microgravity (0g group) for 1.5, 6, 12 and 24 h. mRNA levels of different genes that encode the components of both tubulin and actin cytoskeleton were assessed by qRT-PCR. In the 0g group the mRNA levels of beta-tubulin and Msps were reduced after 1.5 h of the exposure and remained unchanged until 12 h, while they exceeded the control level after 24 h. The mRNA level of chaperonin containing T-complex 1 polypeptide subunits recovered earlier: after 6 and 12 h of the microgravity exposure. At the same time, the hypergravity effect led to more significant changes in the mRNA level of TCP1 complex components compared with those of tubulin and Msps. The mRNA level of beta-actin isoforms under micro- and hypergravity was decreased up to 12 h of the exposure, however, it remained reduced under microgravity conditions, while it recovered (Act87E) and even exceeded (Act57B) the reference level under hypergravity conditions. The mRNA level of supervillin was almost unchanged. Under microgravity conditions the mRNA level of fimbrin was decreased (it recovered by the 24 h time point), while the mRNA level of alpha-actinin was significantly increased by the 12 h time point of the exposure and after 24 h it was reduced to the control level. In contrast, under hypergravity conditions the mRNA level of fimbrin initially increased, and after 24 h it dropped below the control, while the mRNA level of alpha-actinin was significantly reduced, and after 24 h it was higher than the reference level. Similar results were obtained earlier in the experiments in rodents, but similar dynamics were observed for alpha-actinin isoforms 1 and 4, although no changes were observed for fimbrin. Since Drosophila melanogaster has no alpha-actinin isoform 4, it is hypothesized that its role in the cell is played by fimbrin.
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Публикация |
Analysis of the expression levels of genes that encode cytoskeletal proteins in Drosophila melanogaster larvae during micro- and hypergravity effect simulations of different durations
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Огнева И. В. (Профессор)
Свистунов А.А (Первый проректор)
Несвижский Юрий Владимирович (Профессор)
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Biophysics (Russian Federation) |
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The goal of this study was to find genes that encode cytoskeletal proteins that are potential candidates for the role of triggers in cell mechanosensitivity in the fruit fly. Centrifugation was used to simulate the hypergravity effects (2g group); the constantly changing orientation of the larvae in the gravity field was performed in order to simulate the effects of microgravity (0g group) for 1.5, 6, 12 and 24 h. mRNA levels of different genes that encode the components of both tubulin and actin cytoskeleton were assessed by qRT-PCR. In the 0g group the mRNA levels of beta-tubulin and Msps were reduced after 1.5 h of the exposure and remained unchanged until 12 h, while they exceeded the control level after 24 h. The mRNA level of chaperonin containing T-complex 1 polypeptide subunits recovered earlier: after 6 and 12 h of the microgravity exposure. At the same time, the hypergravity effect led to more significant changes in the mRNA level of TCP1 complex components compared with those of tubulin and Msps. The mRNA level of beta-actin isoforms under micro- and hypergravity was decreased up to 12 h of the exposure, however, it remained reduced under microgravity conditions, while it recovered (Act87E) and even exceeded (Act57B) the reference level under hypergravity conditions. The mRNA level of supervillin was almost unchanged. Under microgravity conditions the mRNA level of fimbrin was decreased (it recovered by the 24 h time point), while the mRNA level of alpha-actinin was significantly increased by the 12 h time point of the exposure and after 24 h it was reduced to the control level. In contrast, under hypergravity conditions the mRNA level of fimbrin initially increased, and after 24 h it dropped below the control, while the mRNA level of alpha-actinin was significantly reduced, and after 24 h it was higher than the reference level. Similar results were obtained earlier in the experiments in rodents, but similar dynamics were observed for alpha-actinin isoforms 1 and 4, although no changes were observed for fimbrin. Since Drosophila melanogaster has no alpha-actinin isoform 4, it is hypothesized that its role in the cell is played by fimbrin.
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Публикация |