Activation Of Anaerobic Glycolysis Research Articles

Exercise Training Decreases Nitrite Concentration in the Heart and Locomotory Muscles of Rats Without Changing the Muscle Nitrate Content.

Skeletal muscles are postulated to be a potent regulator of systemic nitric oxide homeostasis. In this study, we aimed to evaluate the impact of physical training on the heart and skeletal muscle nitric oxide bioavailability (judged on the basis of intramuscular nitrite and nitrate) in rats. Rats were trained on a treadmill for 8 weeks, performing mainly endurance running sessions with some sprinting runs. Muscle nitrite (NO2-) and nitrate (NO3-) concentrations were measured using a high-performance liquid chromatography-based method, while amino acids, pyruvate, lactate, and reduced and oxidized glutathione were determined using a liquid chromatography coupled with tandem mass spectrometry technique. The content of muscle nitrite reductases (electron transport chain proteins, myoglobin, and xanthine oxidase) was assessed by western immunoblotting. We found that 8 weeks of endurance training decreased basal NO2- in the locomotory muscles and in the heart, without changes in the basal NO3-. In the slow-twitch oxidative soleus muscle, the decrease in NO2- was already present after the first week of training, and the content of nitrite reductases remained unchanged throughout the entire period of training, except for the electron transport chain protein content, which increased no sooner than after 8 weeks of training. Muscle NO2- level, opposed to NO3-, decreases in the time course of training. This effect is rapid and already visible in the slow-oxidative soleus after the first week of training. The underlying mechanisms of training-induced muscle NO2- decrease may involve an increase in the oxidative stress, as well as metabolite changes related to an increased muscle anaerobic glycolytic activity contributing to (1) direct chemical reduction of NO2- or (2) activation of muscle nitrite reductases.

Features of the respiratory burst state of neutrophils and the activity of NAD(P)-dependent dehydrogenases in patients with widespread purulent peritonitis in the prognosis of the development of sepsis

Aim: Studying of the respiratory burst state and NAD(P)-dependent dehydrogenases activities in blood neutrophils features in the forecast of the development of abdominal sepsis in patients with widespread purulent peritonitis (WPP).
 Materials and methods. The study involved 50 patients with WPP in the preoperative period. Abdominal sepsis was developed by 35 patients (70.0%) from 5 to 10 days postoperative period, 15 patients (30.0%) hadn’t complications. The respiratory burst condition of blood neutrophils was examined using a chemiluminescent assay. Intracellular activity of the NAD- and NADP-dependent dehydrogenases was researched with using bioluminescent methods.
 Results. It was found that patients with WPP whose dynamics of the preoperative period will develop sepsis the chemiluminescent activity of blood neutrophils was characterized by a reduced level of spontaneous synthesis of the primary reactive oxygen species (ROS) and elevated levels of spontaneous synthesis of secondary ROS relative of the indicators identified in patients without subsequent complications. The feature of neutrophil metabolism in WPP patients without subsequent development of sepsis was high activity of the anaerobic lactate dehydrogenase reaction and decrease in activity of the NADP-dependent decarboxylating malate dehydrogenase. In patients with WPP and the subsequent development of sepsis was found high level of NAD-dependent substrates outflow citric acid cycle in the reaction of amino acid metabolism via glutamate dehydrogenase that may affect the activity of aerobic respiration in the neutrophils. Using correlation analysis was found that the intensity of the neutrophils respiratory burst in patients with no subsequent complications depends on the activity of anaerobic glycolysis.
 Conclusion. The established differences in the state of the respiratory burst and the activity of enzymes in neutrophils in patients with WPP, in depending on the subsequent development of the sepsis, determine the possibility of developing the method of forecasting complications and developing immunoactive therapy in the postoperative period of WPP.

Integrated mRNA-Seq and miRNA-Seq analysis reveals the microRNAs-transcription factor-mRNA regulatory network of pearl gentian grouper under acute hypoxia

Pearl gentian grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀) is a hybrid fish with high commercial value and it has the highest yield among groupers farmed along the southeastern coast of China. However, hypoxia is not only a key factor to limit the healthy growth and survival of the grouper under high-density and intensive farming, but also hinders the rapid development of its aquaculture industry. In this study, we analyzed the transcriptomic characteristics of pearl gentian grouper under acute hypoxic stress (DO 0.5 mg/L) for 1 h, 3 h, 6 h, and 9 h and normoxic condition (DO 5.5 mg/L) to determine the biological processes and molecular regulatory mechanisms underlying the response to hypoxia. By an integrated analysis of mRNA and miRNA sequencing data, we constructed a microRNA-transcription factor-mRNA regulatory network containing 52 miRNAs, 17 transcription factors, and 72 genes. During acute hypoxic stress, these key molecules were involved in several important biological processes. The activation of anaerobic glycolysis and fatty acid metabolism, cell cycle arrest contributed to grouper survival under hypoxia. Based on analyses of gene expression profiles, hypoxic stress for 3–6 h and 9 h were identified as two critical periods with significantly altered physiologic and metabolic responses in pearl gentian grouper (e.g., the transformation of energy metabolism patterns and severe oxidative stress). These results indicate that intervention measures should be implemented within 6 h of acute hypoxia in high-density and intensive farming of the grouper. Thus, our study contributes to a deeper understanding of the molecular regulatory mechanism of pearl gentian grouper adaption to acute hypoxia and provide an important scientific basis for the management of hypoxia in high-density and intensive farming practices.

Functional Magnetic Resonance Spectroscopy of Lactate in Alzheimer Disease: A Comprehensive Review of Alzheimer Disease Pathology and the Role of Lactate.

Functional 1H magnetic resonance spectroscopy (fMRS) is a derivative of dynamic MRS imaging. This modality links physiologic metabolic responses with available activity and measures absolute or relative concentrations of various metabolites. According to clinical evidence, the mitochondrial glycolysis pathway is disrupted in many nervous system disorders, especially Alzheimer disease, resulting in the activation of anaerobic glycolysis and an increased rate of lactate production. Our study evaluates fMRS with J-editing as a cutting-edge technique to detect lactate in Alzheimer disease. In this modality, functional activation is highlighted by signal subtractions of lipids and macromolecules, which yields a much higher signal-to-noise ratio and enables better detection of trace levels of lactate compared with other modalities. However, until now, clinical evidence is not conclusive regarding the widespread use of this diagnostic method. The complex machinery of cellular and noncellular modulators in lactate metabolism has obscured the potential roles fMRS imaging can have in dementia diagnosis. Recent developments in MRI imaging such as the advent of 7 Tesla machines and new image reconstruction methods, coupled with a renewed interest in the molecular and cellular basis of Alzheimer disease, have reinvigorated the drive to establish new clinical options for the early detection of Alzheimer disease. Based on the latter, lactate has the potential to be investigated as a novel diagnostic and prognostic marker for Alzheimer disease.

Glycolysis process activation in preserved red blood cells by nanotechnological treatment of resuspending solutions

Currently, the use of nanotechnology opens up new opportunities to influence the processes of anaerobic glycolysis and the activity of hexose monophosphate reactions in preserved erythrocytes. Components containing donor red blood cells on CPDA-1 preservative were examined. Modified solutions of 0.9% NaCl and with 5% glucose were used as resuspending solutions. The solutions were treated with magnetite nanoparticles (ICNB brand) by the Belousov method. The amounts of 2,3-DPG, ATP, reduced glutathione, and glutathione peroxidase were determined by spectrophotometry. This study opens up new possibilities for increasing the shelf life and functional activity of preserved erythrocytes. The study showed a reliable increase in ATP and reduced glutathione, a decrease in 2,3-DPG and glutathione peroxidase. It was found that the activation of anaerobic glycolysis was less pronounced in tests with modified physiological saline than in tests with glucose solution. On the contrary, the pentose glucose oxidation cycle prevailed. A comprehensive analysis of the data obtained indicates the membrane-protective effect of the modified resuspending solutions. The membrane-protective effect is due to an increase in ATP and reduced glutathione, which ensures the redox potential of the cell in an equilibrium state. Magnetite nanoparticles (ICNB) change the mobility and orientation of hydrogen protons in resuspending solutions. This polarizes the aqueous sector of the erythrocyte microenvironment due to van der Waals forces, which is the main reason for activation of ATP phosphate residue hydrolysis and switching of intracellular enzymes regulating anaerobic glycolysis and pentose phosphate cycle into the active state. As a result, transmembrane metabolism and metabolism change, the energy state of erythrocytes changes, and enzymes are activated. All this has a significant impact on the energy supply of preserved red blood cells and preservation of their functional activity under storage conditions at 2 to 6 ºС.

Physiological response and biomarkers in kickboxing - systematic review

Kickboxing is a relatively young sport with growing popularity across the World. There is a rapidly growing need for clear and valid detailed specific information about the effects of kickboxing as an activity. Therefore, this research aimed to collect all legitimate evidence about the acute and chronic physiological response, as well as monitoring and utilization of specific biomarkers in kickboxing. For this systematic review, the authors conducted research in the three relevant scientific bases and one search engine for this specific topic, PubMed, Web of Science, Scopus, and Google Scholar. After title and abstract screening, and review of full-text manuscripts regarding inclusion criteria, 16 studies were included in the review. Studies were sorted according to the type of research methods used. Results revealed a positive chronic impact of kickboxing on the overall athletes' health, speaking of hormonal, blood, and immune parameters. Although, the acute reaction of athletes' organisms includes the increase in levels of cortisol, growth hormone, testosterone, glucose, indicators of lipid peroxidation activity, H+, BEecf, lactic acid, and pyruvic acid, while the levels of myostatin and irisin tend to decrease. Such results confirm kickboxing as a high-demand anaerobic glycolytic activity. Secondly, the currently available analyses of the greatest value for monitoring in competitive kickboxing are hormonal, especially for testosterone & cortisol, immunoassays & saliva assays for immunity tracking, and genetic analyses as auxiliary tools. Further investigations should include females, kickboxing discipline differentiation, weight and age stratification, and differentiation.

The Significance of COVID-19 Diseases in Lipid Metabolism Pregnancy Women and Newborns

Coronavirus disease (COVID-19) is an infectious disease caused by SARS-CoV-2. Elderly people, people with immunodeficiency, autoimmune and malignant diseases, as well as people with chronic diseases have a higher risk of developing more severe forms of the disease. Pregnant women and children can becomesick, although more often they are only the carriers of the virus. Recent studies have indicated that infants can also be infected by SARS-CoV-2 and develop a severe form of the disease with a fatal outcome. Acute Respiratory Distress Syndrome (ARDS) ina pregnant woman can affect the supply of oxygen to the fetus and initiate the mechanism of metabolic disorders of the fetus and newborn caused by asphyxia. The initial metabolic response of the newborn to the lack of oxygen in the tissues is the activation of anaerobic glycolysis in the tissues and an increase in the concentration of lactate and ketones. Lipid peroxidation, especially in nerve cells, is catalyzed by iron released from hemoglobin, transferrin and ferritin, whose release is induced by tissue acidosis and free oxygen radicals. Ferroptosis-inducing factors can directly or indirectly affect glutathione peroxidase through various pathways, resulting in a decrease in the antioxidant capacity and accumulation of lipid reactive oxygen species (ROS) in the cells, ultimately leading to oxidative cell stress, and finally, death. Conclusion: damage to the mitochondria as a result of lipid peroxidation caused by the COVID-19 disease can cause the death of a newborn and pregnant women as well as short time and long-time sequelae.

Mitochondrial Adaptations of Obese Adult Rat Visceral Adipocytes which Differ in Nutritional Status during Childhood

Mitochondria are highly dynamic organelles that can adapt to different environmental stimuli. This study aims to analyze differences in the function and biogenesis of mitochondria and anaerobic glycolysis activity between obese adult rats that were undernourished in childhood (LCD28) and those which were normal (SD28) or already fat (HFD28). Sprague-dawley rats were assigned groups and given diet treatments from December 2017 until August 2018 in the Laboratory Animal Management Unit, Faculty of Veterinary Medicine, Bogor Agricultural Institute. To observe mitochondrial function and biogenesis, MnSOD activity and PGC1α levels were analyzed using ELISA. To investigate the existence of anaerobic glycolysis, LDH activity was analyzed using a spectrophotometry method. The results showed that MnSOD activity and PGC1α levels of LCD28 increased compared to the other groups, signifying higher mitochondrial function and biogenesis. Meanwhile, no significant difference in LDH activity was found, signifying similar levels of anaerobic glycolysis.

Дикарбонільний стрес: гіпотеза клітинного пошкодження в умовах гіпоксії. Пусковий механізм розвитку мультиорганної дисфункції

Актуальність. Різноманітні критичні стани організму часто асоційовані з розвитком гіпоксії, у результаті чого активуються механізми гліколізу, під впливом звільнення стресових гормонів розвивається гіперглікемія. Показано, що в умовах анаеробного гліколізу на тлі гіперглікемії в клітинах продукуються токсичні речовини, що викликають гліколізування протеїнів та нуклеїнових кислот. Разом із порушенням функції клітинних білків виникає мітохондріальна дисфункція, що призводить до енергетичного дефіциту та порушення функціонування органів. Мета дослідження: виявити провідні механізми дикарбонільного стресу, їх значення для формування критичних станів організму та визначити найбільш перспективні способи корекції для фахівців у галузі інтенсивної терапії. Матеріали та методи. Детальне вивчення результатів сучасних наукових досліджень процесів вуглеводного обміну при патологічних станах на підставі інформації, поданої в Інтернеті. Результати. Провідна роль у пошкодженні клітинних структур організму при дикарбонільному стресі належить гліоксалю та метилгліоксалю. Ці хімічні сполуки утворюються як побічні продукти анаеробного гліколізу. Збільшенню їх синтезу сприяють активація анаеробного гліколізу та гіперглікемія. Дикарбонільні сполуки вступають у хімічні реакції з амінними групами протеїнів, нуклеїнових кислот та інших біологічно активних сполук, порушуючи їх функціонування. Природна детоксикація здійснюється гліоксалазною системою за участю відновленого глутатіону, що є основним компонентом антиоксидантної системи. Прогресування окислювального стресу та поява антиоксидантного дефіциту зумовлюють збільшення тяжкості пошкоджень, що викликає підвищене утворення гліоксалю та метилгліоксалю. Профілактика дикарбонільного стресу здійснюється за допомогою збільшення потужності антиоксидантної системи, насамперед за рахунок зростання утворення відновленого глутатіону. Для нейтралізації токсичних дикарбонільних метаболітів можуть бути використані препарати, що виконують функцію «пасток». Перспективне застосування терапії, що спрямована на усунення мітохондріальної дисфункції. Висновки. Нова проблема пошкодження організму в умовах дикарбонільного стресу диктує необхідність аналізу та переоцінки численної кількості заходів інтенсивної терапії. Докладне вивчення особливостей вуглеводного обміну при різних критичних станах, включаючи визначення концентрації гліоксалю та метилгліоксалю, моніторинг рівня глікемії та кліренсу лактату в поєднанні з поверненням до оцінювання стану компенсації функції прооксидантної/антиоксидантної системи організму, є одним із перспективних напрямів наукових досліджень у клініці, що має розпочатися. Фахівці з інтенсивної терапії щоденно стикаються із ситуаціями, коли дикарбонільний стрес може виступати як один із дійсних механізмів формування органної та мультиорганної дисфункції й визначає розвиток декомпенсації життєво важливих функцій. Навчитися протистояти йому є актуальною найближчою задачею.

Identification of Candidate Genes Associated With Hypoxia Tolerance in Trachinotus blochii Using Bulked Segregant Analysis and RNA-Seq.

Golden Pompano (Trachinotus blochii) has rapidly developed into the one of the main valuable fish species in Chinese marine aquaculture. Due to its rapid growth, active metabolism, and high oxygen consumption, hypoxia will increase its mortality and cause serious economic losses. We constructed two experimental groups of fish with different degrees of tolerance to hypoxia, used BSR-Seq analysis based on genome and genetic linkage groups to locate SNPs and genes that were related to the differences in hypoxia tolerance. The results showed that hypoxia tolerance SNPs of golden pompano may be jointly determined by multiple linkage groups, especially linkage groups 18 and 22. There were 768 and 348 candidate genes located in the candidate regions of the brain and liver, respectively. These genes were mainly involved in anaerobic energy metabolism, stress response, immune response, waste discharge, and cell death. The prostaglandin-endoperoxide synthase 2 (PTGS2) on LG8, which is involved in the metabolism of arachidonic acid, has a G/A nonsynonymous mutation at position 20641628, and the encoded amino acid was changed from hydrophobic aspartic acid to asparaginate. The specific pathway of the RIG-I-like receptor signaling pathway in the liver may mediate the metabolic system and the immune system, linking glucose metabolism with immune regulation. The death of the hypoxia-intolerant group may be due to the accumulation of lactic acid caused by the activation of anaerobic glycolysis during the early stage of hypoxia stress, and the activation of type I interferon was inhibited, which resulted in decreased immunity. Among the genes involved in the RIG-I-like receptor signaling pathway, the CYLD Lysine 63 Deubiquitinase (CYLD) located on LG16 had a G/T nonsynonymous mutation at position 13629651, and the encoded amino acid was changed from alanine acid to valine. The interferon induced with helicase C domain 1 (Ifih1) located on LG18 has a G/C nonsynonymous mutation at position 16153700, and the encoded hydrophilic glycine was changed to hydrophobic alanine. Our findings suggest these SNPs may assist in the molecular breeding of hypoxia-tolerant golden pompano, and speculate that the balance of glucose and lipid metabolism plays a key role in Trachinotus blochii under acute hypoxia.

Mechanisms of glycolysis and bioenergy state of cells at the stages of cardiosurgical interventions in patients with hypoxic — ischemic impairments of the brain

Objective — to study the features of bioenergetic provision of oxidative homeostasis (OH) in patients with hypoxic‑ischemic brain lesions (HIBL) before and after cardiac surgery (CS) using artificial circulation (AC).
 Methods and subjects. Clinical and biochemical studies were performed in 38 patients, including 14 with ischemic stroke, 15 with encephalopathy, and 9 with severe cognitive dysfunction.
 Results. Analysis of metabolic indicators of glycolysis activity and energy homeostasis of cells before and after CS revealed the patterns of changes in the disorganization of glycolysis mechanisms, intensification of anaerobic mechanisms while limiting the energy supply of cells. The obtained data confirm the formation of specific postoperative metabolic provision of bioenergy in patients with CS, which should be considered as one of the triggers of HIBL and individualization of antioxidant cerebroprotection in the preoperative period, taking into account the state of bioenergetic metabolism of cells and the dominant mechanisms of glycolysis.
 Conclusions. Preoperative antioxidant cerebroprotection as a means of prevention of hypoxic‑ischemic brain lesions during cardiac surgery using artificial circulation should be based on the determination of bioenergetic and metabolic reserves, the depletion of which by antioxidant drugs suppression should not be considered, as activation of anaerobic glycolysis at simultaneous metabolic suppression of mitochondrial bioenergetics is a factor of formation or aggravation of ischemic lesions of brain.

Effect of Type 2 Diabetes Mellitus on the Hypoxia-Inducible Factor 1-Alpha Expression. Is There a Relationship with the Clock Genes?

Limited reports exist on the relationships between regulation of oxygen homeostasis and circadian clock genes in type 2 diabetes. We examined whether the expression of Hypoxia-Inducible Factor-1α (HIF-1α) and HIF-2α relates to changes in the expression of clock genes (Period homolog proteins (PER)1, PER2, PER3, Retinoid-related orphan receptor alpha (RORA), Aryl hydrocarbon receptor nuclear translocator-like protein 1 (ARNTL), Circadian locomotor output cycles kaput (CLOCK), and Cryptochrome proteins (CRY) 1 and CRY2) in patients with type 2 diabetes. A total of 129 subjects were evaluated in this cross-sectional study (48% with diabetes). The gene expression was measured by polymerase chain reaction. The lactate and pyruvate levels were used as surrogate of the hypoxia induced anaerobic glycolysis activity. Patients with diabetes showed an increased plasma concentration of both lactate (2102.1 ± 688.2 vs. 1730.4 ± 694.4 uM/L, p = 0.013) and pyruvate (61.9 ± 25.6 vs. 50.3 ± 23.1 uM/L, p = 0.026) in comparison to controls. However, this finding was accompanied by a blunted HIF-1α expression (1.1 (0.2 to 5.0) vs. 1.7 (0.4 to 9.2) arbitrary units (AU), p ≤ 0.001). Patients with diabetes also showed a significant reduction of all assessed clock genes’ expression. Univariate analysis showed that HIF-1α and almost all clock genes were significantly and negatively correlated with HbA1c concentration. In addition, positive correlations between HIF-1α and the clock genes were observed. The stepwise multivariate regression analysis showed that HbA1c and clock genes independently predicted the expression of HIF-1α. Type 2 diabetes modifies the expression of HIF-1α and clock genes, which correlates with the degree of metabolic control.

Dexmedetomidine alleviated neuropathic pain in dorsal root ganglion neurons by inhibition of anaerobic glycolysis activity and enhancement of ROS tolerance.

Neuropathic pain is a kind of chronic pain that is triggered or caused primarily by damage to the nervous system and neurological dysfunction. It’s known that dexmedetomidine is a new type of highly selective alpha2-adrenoceptor agonist with sedation, anti-anxiety, analgesic and other effects. However, the function and mechanism of dexmedetomidine on neuropathic pain are not clear. Rat DRG neurons were isolated and identified using immunofluorescence assay. Following treatment with H2O2, dexmedetomidine or ROS inhibitor (NAC), the apoptosis and ROS levels were examined by flow cytometery; apoptosis- and anaerobic glycolysis-related proteins were determined by Western blot assay; glucose consumption, pyruvic acid, lactic acid and ATP/ADP ratios were also measured. The results revealed that dexmedetomidine inhibited H2O2-induced apoptosis and reactive oxygen species (ROS) in rat DRG neurons and in addition, dexmedetomidine down-regulated the expression levels of anaerobic glycolysis-related proteins, significantly reduced glucose, pyruvic acid and lactic acid levels. It also increased the ATP/ADP ratio in H2O2-treated rat dorsal root ganglion (DRG) neurons. Moreover, we also demonstrated that ROS inhibitor (NAC) also inhibited H2O2-induced apoptosis and anaerobic glycolysis in rat DRG neurons. In conclusion, dexmedetomidine suppressed H2O2-induced apoptosis and anaerobic glycolysis activity by inhibiting ROS, in rat DRG neurons. Therefore, dexmedetomidine might play a pivotal role in neuropathic pain by the inhibition of ROS.

A transgenic mouse expressing miR-210 in proximal tubule cells shows mitochondrial alteration: possible association of miR-210 with a shift in energy metabolism.

Previously we reported that the microRNA miR-210 is aberrantly upregulated in clear cell renal cell carcinoma (ccRCC) via deregulation of the VHL-HIF pathway. In the present study, to investigate the biological impact of miR-210 in ccRCC tumorigenesis, we developed a transgenic mouse line expressing miR-210 in proximal tubule cells under control of the mouse SGLT2/Slc5a2 promoter. Light microscopy revealed desquamation of the tubule cells and regeneration of the proximal tubule, suggesting that miR-210 expression led to damage of the proximal tubule cells. Electron microscopy revealed alterations to the mitochondria in proximal tubule cells, with marked reduction of the mitochondrial inner membrane, which is the main site of ATP production via oxidative phosphorylation (OxPhos). An additional in vitro study revealed that this loss of the inner membrane was associated with downregulation of Iscu and Ndufa4, the target genes of miR-210, suggesting that the miR-210-ISCU/NDUFA4 axis may affect mitochondrial energy metabolism. Furthermore, metabolome analysis revealed activation of anaerobic glycolysis in miR-210-transfected cells, and consistent with this the secretion of lactate, the final metabolite of anaerobic glycolysis, was significantly increased. Lactate concentration was higher in the kidney cortex of transgenic mice relative to wild-type mice, although the difference was not significant (p = 0.070). On the basis of these findings, we propose that miR-210 may induce a shift of energy metabolism from OxPhos to glycolysis by acting on the mitochondrial inner membrane. In addition to activation of glycolysis, we observed activation of the pentose phosphate pathway (PPP) and an increase in the total amount of amino acids in miR-210-transfected cells. This may help cells synthesize nucleotides and proteins for building new cells. These results suggest that miR-210 may be involved in the metabolic changes in the early stage of ccRCC development, helping the cancer cells to acquire growth and survival advantages. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

EFFECTIVENESS OF CORRECTION OF DISCUSSION OF CARBOHYDRATE METABOLISM IN RATS WITH CRANIOCEREBRAL INJURY

The development and introduction into practice of emergency neurology of new drugs capable of influencing secondary neuronal damage in patients with traumatic brain injury (TBI) has made a significant impact on the recovery of such patients. Aim of the work was to evaluate the magnitude of cerebroprotective action of amantadine sulfate in terms of impaired carbohydrate metabolism of the brain against the background of experimental TBI. Materials and methods. The experimental model of severe TBI was caused by the action of a carbon dioxide flow under pressure, which was created using a gas balloon of pneumatic gun. The therapeutic effect of amantadine sulfate on TBI model was evaluated with a 5 mg/kg dose. The pseudo-operated animals and control group received 0.9 % NaCl solution at a dose of 2 ml/kg with IV administration. Results. The obtained results confirm the formation of a hypo-energetic state in the brain cells of rats on the background of TBI, which is probably a consequence of the suppression of the processes of oxidative phosphorylation and its conjugation with tissue respiration. TBI is accompanied by activation of anaerobic glycolysis and inhibition of aerobic glucose oxidation, proving that there is an imbalance between the levels of lactate and pyruvate intermediates. 8-day infusion of rats with traumatic brain injury of amantadine sulfate, inhibited anaerobic glycolysis hyperactivation, stimulated tissue respiration processes, reduced signs of lactic acidosis and development of secondary alteration of brain cells by non-oxidized products. Conclusions. In the course of the experiment, it was found that the treatment of rats, with TBI model, with 0.9 % NaCl solution did not significantly contribute to the restoration of impaired energy metabolism (p<0.05) and did not reduce the manifestation of metabolic acidosis in the damaged brain (p<0.05). The use of amantadine sulphate in TBI rats significantly corrects carbohydrate metabolism disorders than 0.9 % NaCl solution and contributes to the reduction of metabolic acidosis in the damaged brain (p<0.05).

Hypoxic syndrome in hereditary diseases of connective tissue

Objective. To analyze the influence of clinical and biochemical markers on the diagnostic algorithm for hypoxic syndrome in children with genetic diseases of connective tissue.Characteristics of children and methods. There were examined 60 children were examined: 50 children with Ehlers–Danlossyndrome and 10 children with Marfan syndrome.Results. Constant fatigue, fast fatigue, low tolerance to physical exercises, pale skin, muscle hypotonia and headache are the most frequent symptoms of a hypoxic state in children with connective tissue dysfunction. At the biochemical level, hypoxic syndrome was characterized by signs of oxidative stress. The authors noted an decrease in pO2 , an increase in pCO2 in the circulating blood, an increase in the activity of anaerobic glycolysis with accumulation of lactate and pyruvate, a decrease in the synthesis of macroergs (ATP, ADP, AMP) and the activity of the mitochondrial respiratory enzymes, the emersion of mitochondrial insufficiency in the form of ripped red fibers (RRF) in myocytes , conglomerates of lipids, calcium, glycogen. There was an increased content of peroxidation products in the blood, urine and saliva.Conclusion. Periodic use of antihypoxants, drugs with detoxifying and immunomodulatory effect is justified for the treatment of children.

Upregulation of Krebs cycle and anaerobic glycolysis activity early after onset of liver ischemia.

The liver is a highly vascularized organ receiving a dual input of oxygenated blood from the hepatic artery and portal vein. The impact of decreased blood flow on glucose metabolism and how hepatocytes could adapt to this restrictive environment are still unclear. Using the left portal vein ligation (LPVL) rat model, we found that cellular injury was delayed after the onset of liver ischemia. We hypothesized that a metabolic adaptation by hepatocytes to maintain energy homeostasis could account for this lag phase. Liver glucose metabolism was characterized by 13C- and 1H-NMR spectroscopy and analysis of high-energy metabolites. ALT levels and caspase 3 activity in LPVL animals remained normal during the first 12 h following surgery (P<0.05). Ischemia rapidly led to decreased intrahepatic tissue oxygen tension and blood flow (P<0.05) and increased expression of Hypoxia-inducible factor 1-alpha. Intrahepatic glucose uptake, ATP/ADP ratio and energy charge level remained stable for up to 12 h after ligation. Entry of glucose in the Krebs cycle was impaired with lowered incorporation of 13C from [U-13C]glucose into glutamate and succinate from 0.25 to 12 h after LPVL. However, total hepatic succinate and glutamate increased 6 and 12 h after ischemia (P<0.05). Glycolysis was initially reduced (P<0.05) but reached maximum 13C-lactate (P<0.001) and 13C-alanine (P<0.01) enrichments 12 h after LPVL. In conclusion, early liver homeostasis stems from an inherent ability of ischemic hepatocytes to metabolically adapt through increased Krebs cycle and glycolysis activity to preserve bioenergetics and cell viability. This metabolic plasticity of hepatocytes could be harnessed to develop novel metabolic strategies to prevent ischemic liver damage.

THE PHENOTYPE AND METABOLISM RELATIONSHIP OF BLOOD NEUTROPHILS IN PATIENTS WITH WIDESPREAD PURULENT PERITONITIS IN THE POSTOPERATIVE PERIOD DYNAMICS

The aim of the study was to examine the relationship of the phenotype and metabolism of neutrophils in patients with widespread purulent peritonitis (WPP) in the dynamics of the postoperative period. The study involved 27 patients with acute surgical diseases and injuries of abdominal organs complicated by WPP. Blood sampling was performed prior to surgery (pre-operative period) and at 7, 14 and 24 day post-operative period. As controls 67 respect healthy people were examined. Research blood neutrophilic granulocytes phenotype was performed by f low cytometry using a direct immunof luorescence whole peripheral blood. The levels of surface receptor expression was assessed by the mean f luorescence intensity. The NADand NADP-dependent dehydrogenases activity in the blood neutrophils studied using bioluminescence method. It was found that in patients with WPP in the preoperative period in the peripheral blood increased content of CD62L+-, HLA-DR+and CD64+-neutrophils. High levels of CD62L+-cells stored within 24 postoperative days, whereas the amount of HLA-DR+and CD64+-neutrophils on 24 postoperative day is reduced to the level of controls. The dynamics of changes in the content of CD64+-cells in the peripheral blood of patients with WPP corresponds to the expression level of CD64-receptor on the membrane of neutrophilic granulocytes. The metabolism of blood neutrophils in patients with WPP in the preand postoperative period is characterized by high intensity of the substrate stream on the citric acid cycle, low activity of NADP-dependent glutamate dehydrogenase and aerobic reaction of lactate dehydrogenase. In the preoperative period and within 14 days of the postoperative period in neutrophil granulocytes of the patients revealed high activity of anaerobic lactate dehydrogenase reaction characterized by increased activity of anaerobic glycolysis. In the late postoperative period the intensity of anaerobic energy in the neutrophils of patients with WPP was reduced to the control level. The final stage of post-operative treatment the metabolism in the blood neutrophils of patients with WPP is also characterized by the activation of the pentose phosphate cycle, low activity of malate-aspartate shunt mitochondria and high intensity of the substrate interaction between citric acid cycle and reactions of amino acid metabolism. Using correlation analysis set dysregulation between phenotype and the system of intracellular metabolism of neutrophils which can be determined by the migration of activated cells in the inf lammatory focus as well as changes in the activity of intracellular enzymes under different regulatory factors and including postoperative therapy methods peritonitis.

METABOLITHOTROPIC ASPECTS OF CARDIOPROTECTIVE ACTION OF NEW COMBINED MEDICINE BASED ON L-ARGININE AND THIOTRIAZOLIN AT MODELING OF MYOCARDIAL INFARCTION

Objectives: The actual problem of modern medicine is the development of medications in treatment of cardiovascular system diseases. Different combinations of L-arginine with thiotriazolin in acute myocardial ischemia were examined as part of study and the optimal combination has been established - 4:1, named “Argitryl.” The aim is to study the antioxidant, energotropic, and nitric oxide (NO)-modulating mechanisms of the cardioprotective effects of L-arginine with thiotriazolin (4:1) (Argitryl) drug candidate compared to 4.2% solution for injections “Tivortin” (TOV “Yuriya-Farm,” Ukraine) on the model of pituitrin-isadrin infarction of myocardium in laboratory rats.Methods: The study was conducted in accordance with Guidelines of State Expert Center of the Ministry of Health of Ukraine. All manipulations were carried out in accordance with the “European Convention for the protection of vertebrate animals used for experimental and other scientific purposes.” The experiments were performed on 40 white outbred rats weighing 170–180 g. The antioxidant activity of the drugs was assessed by reduction of biochemical marker oxidative stress and an increase in the activity of the antioxidant system of the myocardium.Results: The study of energy metabolism has shown that the modeling of myocardial infarction leads to typical ischemic injuries - decrease in ATP, discoordination of the Krebs cycle, activation of anaerobic glycolysis in the cytosolic fraction of the heart of rats from the control group. Administration of Tivortin to animals does not lead to a significant decrease in ischemic injuries in the energy metabolism of the myocardium. The brightest anti-ischemic effect was observed with the appointment of “Argitryl.”Conclusion: The ascertained braking reaction of oxidative stress compared to the control group was found as a result of administration of Argitryl and Tivortin to animals with pituitrin-isadrin myocardial infarction. It was also found that the administration of Argitryl and Tivortin has a positive effect on the indices of the energy metabolism of the ischemic myocardium and leads to the normalization in conjugated NO system/renewed thiols. Argitryl significantly exceeds Tivortin on the influence of antioxidant, energy-tropic, and NO-modulating mechanisms of cardioprotective and anti-ischemic action on the studied indicators

East Urals Radioactive Trace: Dose-dependent functional-metabolic effects in the myocardium of the pygmy wood mouse (Apodemus uralensis) taking into account population size

The population dynamics, radiometric data and biochemical parameters (concentrations of total lipids, proteins, DNA and RNA, activity of succinate dehydrogenase, glucose phosphate isomerase and catalase, as well as lipid peroxidation level) in the myocardium of the pygmy wood mouse (Apodemus uralensis Pall., 1811) inhabiting the area of the East Urals Radioactive Trace (EURT) were analyzed. The functional-metabolic radiation effects as a result external and internal exposure to 137Cs and 90Sr (unweighted total dose rate 0.04–0.5 mGy/day) are characterized by a reduction in lipid catabolism, mitochondrial oxidation and antioxidant defense, as well as the activation of anaerobic glycolysis as well as the protein-synthesizing and genetic apparatus. The data indicate the low efficiency of cell energy production and allow us to state that compensatory myocardial hypertrophy can improve myocardial contractile function. The level of the functional-metabolic response in pygmy wood mice in the EURT area increased with increasing whole-body radiation dose rate and was more pronounced with a large pygmy wood mouse population size. The harmful effects (cardiac decompensation stage) of synergies resulting from non-radiation and radiation factors manifest after population abundance above 30 ind./100 trap-day and a radiation burden above 0.1 mGy/day.