Phosphocreatine Research Articles

Impaired Skeletal Muscle Mitochondrial Efficiency in Smokers

Introduction Cigarette smoke can directly contribute to the development of skeletal muscle dysfunction. Mitochondrial oxidative capacity, a product of energy production efficiency and oxygen consumption, is important for skeletal muscle health. Whether mitochondrial energy production efficiency and muscle oxygen consumption play a role into skeletal muscle dysfunction in smokers has yet to be investigated. Thus, this study sought to test the hypothesis that mitochondrial oxidative capacity and mitochondrial efficiency are reduced in smokers compared to non-smokers. Methods 30 active smokers (Age: 33 ± 4 yrs, BMI: 30.8 ± 11.7 kg/m2) and 30 demographically matched healthy controls (Age: 33 ± 3 yrs, BMI: 32.5 ± 9.1 kg/m2) participated in this study. Participants self-reported the number of cigarettes smoked per day in the past 30 days. Near infrared spectroscopy (NIRS) was used to assess changes in oxygenated (O2Hb) and deoxygenated (HHb) hemoglobin/myoglobin of the gastrocnemius following 15s of electrical stimulation and multiple rapid arterial occlusions. The phosphocreatine (PCr) recovery rate constant (kPCr), an index of mitochondrial oxidative capacity, was calculated by fitting post-stimulation measurements to a monoexponential curve. Muscle oxygen consumption (mVO2 = abs[([ΔO2Hb – Δ HHb]/2] × 60)/(10 × 1.04) x4] × 22.4/1000) was calculated for each occlusion and the slopes of the total hemoglobin (tHb = O2Hb+HHb) signal following each occlusion were converted to muscle blood flow [mBF = 1/hemoglobin (mmol/L) × ΔtHb/Δtime]. Mitochondrial efficiency (MEFF) was calculated by the dividing the estimated mVO2 needed to achieve kPcr at 100% (mVO2estimated) efficiency by the observed mVO2 [Meff = (mVO2estimated/mVO2observed) × 100]. Results The average cigarettes smoked was 5 ± 3 cigarettes per day. kPcr was significantly (p = 0.022) reduced in smokers (1.99 ± 0.19 min-1) compared to non-smokers (2.54 ± 0.19 min-1). Compared to non-smokers (157.2 ± 45.5%), MEFF was significantly (p = 0.025) attenuated in smokers (46.5 ± 10.1%) and significantly associated with mVO2 (r = -0.409, p = 0.009) and kPcr (r = 0.461, p =0.003). Average mVO2 (p = 0.231) and mBF (p = 0.993)were not significantly different in smokers compared to non-smokers. Conclusion Muscle mitochondrial oxidative capacity and mitochondrial efficiency are reduced in smokers compared to non-smokers. However, skeletal muscle blood flow and oxygen consumption were not different between groups. Taken together, mitochondrial oxidative capacity may be reduced prior to alterations in blood flow and oxygen consumption or delivery in young smokers.

Investigating the effect of trigger delay on cardiac 31P MRS signals

The heart’s geometry and its metabolic activity vary over the cardiac cycle. The effect of these fluctuations on phosphorus (31P) magnetic resonance spectroscopy (MRS) data quality and metabolite ratios was investigated. 12 healthy volunteers were measured using a 7 T MR scanner and a cardiac 31P-1H loop coil. 31P chemical shift imaging data were acquired untriggered and at four different times during the cardiac cycle using acoustic triggering. Signals of adenosine-triphosphate (ATP), phosphocreatine (PCr), inorganic phosphate (Pi) and 2,3-diphosphoglycerate (2,3-DPG) and their fit quality as Cramér-Rao lower bounds (CRLB) were quantified including corrections for contamination by 31P signals from blood, flip angle, saturation and total acquisition time. The myocardial filling factor was estimated from cine short axis views. The corrected signals of PCr and gamma-ATP were higher during end-systole and lower during diastasis than in untriggered acquisitions (P<0.05). Signal intensities of untriggered scans were between those with triggering to end-systole and diastasis. Fit quality of PCr and gamma-ATP peaks was best during end-systole when blood contamination of ATP and Pi signals was lowest. While metabolite ratios and pH remained stable over the cardiac cycle, signal amplitudes correlated strongly with myocardial voxel filling. Triggering of cardiac 31P MRS acquisitions improves signal amplitudes and fit quality if the trigger delay is set to end-systole. We conclude that triggering to end-systole is superior to triggering to diastasis.

Blood lipid profile as a diagnostic marker of acute pancreatitis in dogs

Acute pancreatitis is a common non-communicable pathology in dogs, the untimely diagnosis and treatment of which is lethal (27-58% of cases). One of the most common medical tests for acute pancreatitis is to determine the concentration of triglycerides and other fractions of blood lipids. The purpose of the study was to investigate changes in lipid fractions and other biochemical parameters in dogs with acute pancreatitis. For the experiment, two groups of dogs of 10 individuals each were formed. The control group included healthy animals, the experimental group – dogs with acute pancreatitis. It was found that in dogs with acute pancreatitis, the concentration of triglycerides increased by 67% compared with healthy individuals. Cholesterol and phospholipid concentrations also increased by 23 and 9%, respectively. Inflammatory processes in the pancreas led to the development of cytolytic processes in its acini, as well as hepatocytes of the liver, as indicated by increased activity of indicator enzymes ALT, AST, LF and bilirubin. In addition, in sick dogs, the protein-synthesising function of the liver was impaired: a reduced proportion of albumin was registered against the background of increased total serum protein. Acute pancreatitis leads to a decrease in insulin synthesis, which is manifested by hyperglycaemia – in sick dogs, glucose levels are probably twice as high as in healthy animals. In pancreatitis, the violation of digestion and absorption of nutrients caused by intensive use of high-energy compounds (in particular, creatine phosphate) and an increase in creatinine (3.8 times). At the same time, the catabolism of proteins increased and the products of their intermediate and final metabolism – ammonia and urea – accumulated (by 60 and 57%, respectively), which is a sign of reduced filtration capacity of the kidneys. In addition, in acute pancreatitis within the physiological norm, the concentration of calcium decreased, and inorganic phosphorus – increased

Insight into type 2 diabetes impaired exercising mitochondrial oxidative flux: is it blood flow, mitochondria, or neither?

Insight into type 2 diabetes impaired exercising mitochondrial oxidative flux: is it blood flow, mitochondria, or neither?

Molecular docking and DFT studies of a series of tetrazole derivatives and computational studies of Fe3O4@Phenyl phosphate creatine

The synthesis of tetrazoles by Fe3O4@Phenyl phosphate creatine as a catalyst has been reported previously. In this article, the optimized geometry of the tetrazole derivatives and Fe3O4@Phenyl phosphate creatine have been computed with the help of density functional theory (DFT). The detailed interpretations of HOMO–LUMO and molecular electrostatic potential surfaces on the tetrazole derivatives and Fe3O4@Phenyl phosphate creatine were carried out at the B3LYP/6–31+G(d) level of theory that are useful tools for molecular stability prediction. Finally, molecular docking was used as an effective method to describe the biological activity of tetrazole derivatives.

MUSCLE MASS AND ESTIMATED RENAL FUNCTION IN ATHLETES

Objective: Serum creatinine level is the most common marker used to assess renal function. Because creatinine is a derivative of creatine phosphate, an energy source present in the skeletal muscles, serum creatinine levels are influenced by muscle mass. Elderly people with less muscle mass have low creatinine levels, which may lead to an overestimation of their true renal function. Creatinine levels also tend to be lower in females than males, as females tend to have less muscle mass. Serum cystatin C, a cysteine proteinase inhibitor, is another renal function marker that is produced by every nucleated cell in the body, and thus, unlike creatinine, is thought to be little influenced by muscle mass. Cystatin C level is a sensitive marker of reduction in glomerular filtration rate and suggested to be a better predictive factor for cardiovascular events and death. Design and method: Study subjects were 130 male athletes belonging to the National Institute of Fitness and Sports in Kanoya, Japan. A Smedley-type Hand Dynamometer was used to assess grip strength. Serum creatinine and cystatin C levels were measured by enzyme and latex coagulation methods, respectively. Results: Muscle mass was significantly correlated with both right and left grip strength, indicating that muscle mass measured in the present study represents skeletal muscle mass. Serum creatinine levels and estimated glomerular filtration rates calculated from creatinine (eGFRcre) showed significant correlations with muscle mass, suggesting that creatinine is influenced by muscle mass and that true renal function may be underestimated in athletes. Cystatin C and estimated glomerular filtration rates calculated from cystatin C (eGFRcys) were also significantly correlated with muscle mass. Serum creatinine levels were significantly correlated with serum cystatin C levels. Consistent with this, eGFRcre was significantly correlated with eGFRcys. This suggests that muscle mass influences the assessment of renal function in athletes when cystatin C is used as a marker. Conclusions: The findings in this study suggest that muscle mass of the subject must be considered prior to assessment of renal function using cystatin C in athletes.

Metabolic therapy in cardiology from the perspective of evidence-based medicine

Over the past few decades, various applications of the metabolic drugs have been extensively tested. Most of them affect oxygen-dependent processes, improving cellular metabolism and increasing tissue resistance to hypoxia and ischemia. The most promising candidates include components of the respiratory chain, purine nucleosides, and creatine phosphate which affect glucose oxidation and fatty acid metabolism in the Krebs cycle. This review critically evaluates the most popular drugs of this group (adenosine triphosphate, adenosine-5-monophosphate, creatine phosphate, coenzyme Q10, cytochrome C, adenosine, glucose-insulin-potassium solution, L-carnitine, mildronate, and trimetazidine), which are widely represented on the pharmaceutical market. Of all metabolic drugs, only trimetazidine was included in the European and Russian recommendations for the second-line treatment of stable angina. In most clinical studies, the therapeutic efficacy of metabolic drugs has been evaluated using the surrogate endpoints. Despite being actively advertised and widely used in the clinical practice, metabolic drugs currently do not have a convincing evidence base for affecting prognosis (mortality and/or major adverse cardiovascular events). Further studies in large-scale randomised trials are needed to confirm the beneficial effects of the metabolic drugs in cardiovascular medicine.

Utilizing Dynamic Phosphorous-31 Magnetic Resonance Spectroscopy for the Early Detection of Acute Compartment Syndrome: A Pilot Study on Rats.

Introduction: Disasters, including terrorism and earthquakes, are significant threats to people and may lead to many people requiring rescue. The longer the rescue takes, the higher the chances of an individual contracting acute compartment syndrome (ACS). ACS is fatal if diagnosed too late, and early diagnosis and treatment are essential. Objective: To assess the ability of dynamic phosphorus magnetic resonance spectroscopy (31P-MRS) in the early detection of muscular damage in ACS. Materials and Methods: Six ACS model rats were used for serial 31P-MRS scanning (9.4 Tesla). Skeletal muscle metabolism, represented by the levels of phosphocreatine (PCr), inorganic phosphate (Pi), and adenosine triphosphate (ATP), was assessed. The PCr/(Pi + PCr) ratio, which decreases with ischemia, was compared with simultaneously sampled plasma creatine phosphokinase (CPK), a muscle damage marker. Results: The PCr/(Pi + PCr) ratio significantly decreased after inducing ischemia (from 0.86 ± 0.10 to 0.18 ± 0.06; p < 0.05), while CPK did not change significantly (from 89 ± 29.46 to 241.50 ± 113.28; p > 0.05). The intracellular and arterial pH index decreased over time, revealing significant differences at 120 min post-ischemia (from 7.09 ± 0.01 to 6.43 ± 0.13, and from 7.47 ± 0.03 to 7.39 ± 0.04, respectively). In the reperfusion state, the spectra and pH did not return to the original values. Conclusions: The dynamic 31P-MRS technique can rapidly detect changes in muscle bioenergetics. This technique is a promising non-invasive method for determining early muscular damage in ACS.

Development of a novel biosensor for Creatine Kinase (CK-MB) using Surface Plasmon Resonance (SPR)

In this paper, we report a new biosensor for detection of the cardiac biomarker Creatine Kinase (CK-MB) using Surface Plasmon Surface (SPR) technique, with the purpose of the diagnosis of Acute Myocardial Infarction (AMI). For this, we employed gold nanoparticles (AuNP) stabilized with Cysteamine (Cys), and functionalized with Creatine Phosphate (Pcrea), which promote an increase in the sensitivity and accuracy for creatine kinase (CK-MB) detection in saliva and urine samples. The characteristics of the electronic transfer of the sensor could be observed by Electrochemical Impedance Spectroscopy (EIS) and Cyclic Voltammetry (CV), that showed significant redox peaks on electrode surface evidencing the high electron exchange process and low resistance charge transfer between the electrolyte solution and the modified surface. The morphological characteristics were confirmed by Atomic Force Microscopy (AFM), micro-Raman and Contact Angle (CA), as well as the adsorption and interaction processes between the materials, which could be observed by Surface Plasmon Resonance (SPR). The SPR analytical curve, ranged from 5.0 ng mL−1 to 100.0 ng mL−1, provided some important parameters, such as limit of detection (LOD) of 0.209 ng mL−1, limit of quantification (LOQ) of 0.696 ng mL−1, and sensitivity of 8.67 × 10–3 ± 0.17 × 10–3 (°) mL ng−1. The specific interaction between functionalized substrate and CK-MB demonstrates a high affinity of 15.14 ng mL−1 based on the Michaelis-Menten fitting.

Efeitos da atividade física sobre a doença pulmonar obstrutiva crônica: uma revisão de literatura

Introdução: Pacientes com Doença Pulmonar Obstrutiva Crônica (DPOC) apresentam aumento da secreção de muco nas vias aéreas e hipertrofia das células produtoras de muco, levando à obstrução das vias aéreas, aumento da resistência das vias aéreas, limitação ao fluxo aéreo, aprisionamento aéreo e aumento do volume residual, diminuindo a eficiência do diafragma e reduzindo a capacidade para o exercício. Objetivo: O presente estudo tem por objetivo apresentar os benefícios de diferentes tipos de treinamento físico para pacientes com DPOC, através de uma revisão bibliográfica. Métodos e resultados: Para atingir este objetivo, este estudo utilizou trinta de nove artigos e dois livros. As pesquisas foram realizadas na Biblioteca Virtual em Saúde (BVS) com as bases de dados SciELO, PUBMED, e LILACS. As palavras-chaves utilizadas para estas pesquisas, foram: DPOC, Manifestações Sistêmicas, Exacerbações, Fisioterapia, Treinamento Físico Aeróbico e Resistido. Os critérios de inclusão foram artigos publicados a partir do ano de 2006 e com o tema pesquisado, os de exclusão foram artigos publicados com período igual e/ou superior a quinze anos. Os artigos escolhidos foram na língua portuguesa e inglesa. Conclusão: Dessa forma respeitando os princípios do treinamento é possível realizar um condicionamento físico e cardiorrespiratório para os pacientes com DPOC através do treinamento físico resistido e aeróbico.

Color Attributes and Glycolytic Energy Metabolites of Meat From Light and Heavy Bovine Feedlot Carcasses Stimulated With Low-Voltage Electricity

This study investigates the influence of timing and duration of low-voltage electrical stimulation (ES) as well as carcass weight, which are important factors affecting muscle glycolysis and meat color attributes. Longissimus samples from 147 cattle, representative of typical South African feedlot cattle, were assigned to 10 treatment groups according to the combination of carcass weight (small or heavy), time of ES (early [7 min] or late [45 min] postmortem), and duration of ES (30 or 60 s). Early ES and longer ES duration (60 s) produced the highest (P &amp;lt; 0.05) meat lightness (L*). The combination of early ES and heavier carcasses produced the highest (P &amp;lt; 0.05) chroma (C*) 2 d postmortem (butchery period). These results were corroborated by the energy metabolites, such that early stimulation produced the highest amount (P &amp;lt; 0.05) of muscle lactate, glucose, and glucose-6-phosphate, as well as the lowest glycogen, creatine phosphate, and adenosine triphosphate early postmortem. This shows that the application of low-voltage ES on heavier carcass is advantageous in terms of meat color, but the advantages diminish as ageing progressed from 2 to 14 d postmortem.

Effects of acute heat stress on salivary metabolites in growing pigs: an analysis using nuclear magnetic resonance-based metabolomics profiling.

Heat stress (HS) causes adverse impacts on pig production and health. A potential biomarker of HS is required to predict its occurrence and thereby better manage pigs under HS. Information about the saliva metabolome in heat-stressed pigs is limited. Therefore, this study was aimed to investigate the effects of acute HS on the saliva metabolome and identify metabolites that could be used as potential biomarkers. Growing pigs (n = 6, 3 boars, and 3 gilts) were raised in a thermal neutral (TN; 25°C) environment for a 5-d adaptation period (CON). After adaptation, the pigs were first exposed to HS (30°C; HS30) and then exposed to higher HS (33°C; HS33) for 24 h. Saliva was collected after adaptation, first HS, and second HS, respectively, for metabolomic analysis using 1H-nuclear magnetic resonance spectroscopy. Four metabolites had significantly variable importance in the projection (VIP > 1; p < 0.05) different levels in TN compared to HS groups from all genders (boars and gilts). However, sex-specific characteristics affected metabolites (glutamate and leucine) by showing the opposite results, indicating that HS was less severe in females than in males. A decrease in creatine levels in males and an increase in creatine phosphate levels in females would have contributed to a protective effect from protein degradation by muscle damage. The results showed that HS led to an alteration in metabolites related to energy and protein. Protection from muscle damage may be attributed to the alteration in protein-related metabolites. However, energy-related metabolites showed opposing results according to sex-specific characteristics, such as sex hormone levels and subcutaneous fat layer. This study had shown that saliva samples could be used as a noninvasive method to evaluate heat-stressed pigs. And the results in this study could be contributed to the development of a diagnostic tool as a noninvasive biomarker for managing heat-stressed pigs.

Особливості психофізіологічних характеристик у кваліфікованих боксерів

Purpose: to study the features of psychophysiological characteristics in qualified boxers with different levels of performance. Material and methods. The study used the following methods: analysis of scientific and methodological information, Internet sources and generalization of practical experience, psychophysiological research methods, methods of mathematical statistics. A dynamometer «Spuderg» designed by M.P. Savchyn was used to assess the special performance. The index of creatine phosphate working capacity of boxers was determined (according to the test of the boxer's strikes fulfillment for 8 seconds at maximum speed). We examined 26 qualified boxers, members of the national team of Kiev at the age of 18-24 years. According to the creatine phosphate working capacity index, all boxers were divided into two conditional groups. The first group with a high level of working capacity (from 200 and more USD) and the group with a low level of special working capacity (below 200 USD). The first group included 12 people, the second - 14 athletes. Results: based on the analysis of scientific and methodological literature, it was revealed that a high level of special performance in qualified boxers is provided by a high lability of the nervous system and the speed of processing of visual information with simultaneous control over spontaneous actions under conditions of motor response. During the study, an assessment was made of: complex sensorimotor reactions (functional mobility of nervous processes; reaction to a moving object), specific perceptions (reaction rate to non-verbal stimuli). Conclusions. The study made it possible to establish that boxers with a high level of special working capacity have a balance between the processes of arousal and inhibition, while boxers with a reduced level of working capacity have a predominance of arousal processes and high values of impulsivity, which indicate the presence of spontaneous, unprepared actions when responding to external irritants. The connection between the quality and speed of perception of visual information and special performance in qualified boxers was also revealed. Keywords: qualified boxers, psychophysiological characteristics, special working capacity, types of control

A Prospective Study: Highlights of Hippocampal Spectroscopy in Cognitive Impairment in Patients with Type 1 and Type 2 Diabetes.

Diabetes mellitus type 1 and 2 is associated with cognitive impairment. Previous studies have reported a relationship between changes in cerebral metabolite levels and the variability of glycemia. However, the specific risk factors that affect the metabolic changes associated with type 1 and type 2 diabetes in cognitive dysfunction remain uncertain. The aim of the study was to evaluate the specificity of hippocampal spectroscopy in type 1 and type 2 diabetes and cognitive dysfunction. Materials and methods: 65 patients with type 1 diabetes with cognitive deficits and 20 patients without, 75 patients with type 2 diabetes with cognitive deficits and 20 patients without have participated in the study. The general clinical analysis and evaluation of risk factors of cognitive impairment were carried out. Neuropsychological testing included the Montreal Scale of Cognitive Dysfunction Assessment (MoCA test). Magnetic resonance spectroscopy (MRS) was performed in the hippocampal area, with the assessment of N-acetylaspartate (NAA), choline (Cho), creatine (Cr), and phosphocreatine (PCr) levels. Statistical processing was performed using the commercially available IBM SPSS software. Results: Changes in the content of NAA, choline Cho, phosphocreatine Cr2 and their ratios were observed in type 1 diabetes. More pronounced changes in hippocampal metabolism were observed in type 2 diabetes for all of the studied metabolites. Primary risk factors of neurometabolic changes in patients with type 1 diabetes were episodes of severe hypoglycemia in the history of the disease, diabetic ketoacidosis (DKA), chronic hyperglycemia, and increased body mass index (BMI). In type 2 diabetes, arterial hypertension (AH), BMI, and patient’s age are of greater importance, while the level of glycated hemoglobin (HbA1c), duration of the disease, level of education and insulin therapy are of lesser importance. Conclusion: Patients with diabetes have altered hippocampal metabolism, which may serve as an early predictive marker. The main modifiable factors have been identified, correction of which may slow down the progression of cognitive dysfunction.

Rapid, -insensitive, dual-band quasi-adiabatic saturation transfer with optimal control for complete quantification of myocardial ATP flux.

PurposePhosphorus saturation‐transfer experiments can quantify metabolic fluxes noninvasively. Typically, the forward flux through the creatine kinase reaction is investigated by observing the decrease in phosphocreatine (PCr) after saturation of γ‐ATP. The quantification of total ATP utilization is currently underexplored, as it requires simultaneous saturation of inorganic phosphate (Pi) and PCr. This is challenging, as currently available saturation pulses reduce the already‐low γ‐ATP signal present.MethodsUsing a hybrid optimal‐control and Shinnar‐Le Roux method, a quasi‐adiabatic RF pulse was designed for the dual saturation of PCr and Pi to enable determination of total ATP utilization. The pulses were evaluated in Bloch equation simulations, compared with a conventional hard‐cosine DANTE saturation sequence, before being applied to perfused rat hearts at 11.7 T.ResultsThe quasi‐adiabatic pulse was insensitive to a >2.5‐fold variation in B1, producing equivalent saturation with a 53% reduction in delivered pulse power and a 33‐fold reduction in spillover at the minimum effective B1. This enabled the complete quantification of the synthesis and degradation fluxes for ATP in 30‐45 minutes in the perfused rat heart. While the net synthesis flux (4.24 ± 0.8 mM/s, SEM) was not significantly different from degradation flux (6.88 ± 2 mM/s, P = .06) and both measures are consistent with prior work, nonlinear error analysis highlights uncertainties in the Pi‐to‐ATP measurement that may explain a trend suggesting a possible imbalance.ConclusionsThis work demonstrates a novel quasi‐adiabatic dual‐saturation RF pulse with significantly improved performance that can be used to measure ATP turnover in the heart in vivo.

The Protective Effect of Cx43 Protein-Mediated Phosphocreatine on Myocardial Ischemia/Reperfusion Injury.

Objectives To verify the protective effect of phosphocreatine on myocardium in an ischemic model and the possible mechanism of action. Methods The model of myocardial ischemia/reperfusion (I/R) was established by the ligation balloon method. 30 SD rats were randomly divided into three groups, n = 10 in each group. Sham operation group: the coronary artery was not blocked and observed for 120 minutes. The ischemia/reperfusion (I/R) group was given ischemia for 30 minutes and ischemia reperfusion for 90 minutes. Phosphocreatine (PCr) group: after 30 minutes of ischemia, the rats were intraperitoneally injected with PCr (200 mg/kg) for 90 minutes. The animal groups of myocardial ischemia/reperfusion model in vitro were the same as those in vivo. The heart was removed by thoracotomy and washed immediately in H-K buffer solution. Then, the heart was installed on the Langendorff instrument. The concentration of PCr perfusion fluid in the PCr group was 10 mmol/L. The changes in coronary blood flow in isolated myocardium were recorded. The heart rate and electrocardiogram were recorded by RM6240BT. At the end of the experiment, myocardial pathological sections and Cx43 immunofluorescence staining were made, and the contents of malondialdehyde (MDA) in myocardial tissue were detected. Results Phosphocreatinine treatment improved the myocardial ischemia model, performance in electrocardiogram (ECG) changes (ST segment apparent), and histological changes (decrease in necrotic myocardial cells, inflammatory cell infiltration, and a reduction in myocardial edema). At the same time, MDA decreased, while coronary blood flow and Cx43 expression significantly improved. Conclusions Phosphocreatine can improve the electrocardiogram and restore histologic changes in ischemic myocardium and coronary blood flow. The postulated mechanism is by inhibiting the generation of free oxygen radicals and restoring the expression of Cx43 protein.

Kinetic analysis of the transphosphorylation with creatine kinase by pressure-assisted capillary electrophoresis/dynamic frontal analysis.

Kinetic reactions of the transphosphorylation with creatine kinase (CK) were individually investigated between creatine (Cr) and creatine phosphate (CrP) by pressure-assisted capillary electrophoresis/dynamic frontal analysis (pCE/DFA). The transphosphorylations are reversible between Cr and CrP, and reverse reactions inevitably accompany in general batch analyses. In pCE/DFA, the kinetic reaction proceeds in a separation capillary and the product is continuously resolved from the substrate zone. Therefore, the formation rate is kept constant at the substrate zone without the reverse reaction, and the product is detected as a plateau signal. This study demonstrates the direct and individual analyses of both the forward and the backward kinetic reactions with CK by pCE/DFA. A plateau signal was detected in the pCE/DFA with ADP or ATP as one of the products on either the forward or the backward reactions. The Michaelis-Menten constants of Km,ATP (from Cr to CrP) and Km,ADP (from CrP to Cr) were successfully determined through the plateau signal. Determined values of Km,ATP and Km,ADP by pCE/DFA were smaller than the ones obtained by the pre-capillary batch analyses. The results agree with the fact that the reverse reaction is excluded in the analysis of the kinetic reactions. The proposed pCE/DFA is useful on individual analyses of both forward and backward kinetic reactions without any interference from the reverse reaction.

To the Mechanism of Adrenaline Damage to the Heart Tissue and the Mechanism of Cardioprotection by Neonatal, Xenogenic, Cardiac Cells. Dynamics of Creatine Phosphate, Lactate and Malondialdehyde

The development of disturbances in energy processes and damaging free radical reactions in various pathological processes, including cardiovascular diseases, are interrelated and lead to a significant deterioration in the course of diseases. Aim of the study . Research of the dynamics of malondialdehyde, creatine phosphate and lactate in the cardiac tissue of rats under experimental adrenaline stress and during its correction with neonatal, xenogenic, cardiac cells. Methods . The experiment was carried out on outbred male rats. Adrenaline damage to the heart was simulated by a single subcutaneous injection of 0.1% adrenaline solution at a dose of 0.5 mg per 100 g of body weight. The first group (37 rats) was injected subcutaneously with adrenaline, the second group (41 rats) – adrenaline and isolated heart cells of a newborn rabbit at a dose of 500 000. The third group included 6 healthy rats. Results . It was found that a spike in the level of malondialdehyde and, accordingly, the activation of free radical processes in adrenaline damage to the heart, occurred during the restructuring of the energy of the heart cell from intense glycolysis to the restoration of active mitochondrial ATP synthesis (which corresponded to the end of the depletion of lactate and creatine phosphate, and the beginning of the restoration of their content in heart cells). The dynamics of MDA sensitively reflected both the activity and the inhibition of oxidative processes in mitochondria, which manifested itself, respectively, both in the form of peaks and in a low level of MDA and corresponded to the interpretation of the dynamics of lactate and creatine phosphate. In the cardiac tissue of rats with transplantation of neonatal, xenogenic cardiac cells, the accumulation of lactate in the early stages of the experiment decreased, the subsequent depletion of the cellular reserves of creatine phosphate and lactate was inhibited, the period of inhibition (non-increase) in MDA was shorter, the subsequent increase in MDA was more moderate than in control animals. Conclusion . The data obtained indicate that neonatal cardiac cells are able to limit the disturbance of aerobic and of anaerobic processes in the heart tissue and promote the restoration of mitochondrial energy processes. Moreover, they contribute to a more efficient restoration of mitochondrial ATP synthesis, accompanied by a smaller burst of damaging free radical processes.

Comparison of Metabolic Alterations in Serum and Milk Whey Between Inactive Ovaries and Estrus Dairy Cows.

Inactive ovaries (IOs) affect the estrus cycle and timed artificial insemination (TAI) efficiency in dairy cows during early lactation. The objective of the experiment was to determine metabolic changes in the serum and milk whey of dairy cows with IO and estrus. Twenty-eight healthy postpartum Holstein cows in similar age, milk production, and body condition were selected at 30 days postpartum for tracking to 70 days postpartum, and estrus performance was recorded through Afi Farm® software. The ovarian status and follicular diameter of dairy cows were examined by an experienced breeder through B-ultrasound and rectal examination. Fourteen normal estrus cows were allocated to control group A and 14 cows with IO to group B, all at 30–70 days postpartum. The serum and milk whey in the two groups of cows at 70 days postpartum were used for non-targeted nuclear magnetic resonance (1H-NMR) analysis to measure the different metabolites of cows with IO. In group B compared with group A at 70 days postpartum, there was an increase in the milk whey of six different metabolites including succinate, creatine phosphate, glycine, myo-inositol, glycolate, and orotate and a decrease in the milk whey of seven metabolites, including alanine, creatinine, o-phosphorylcholine, lactose, taurine, galactose, and glucose-1-phosphate. There was an increase in the serum of group B cows of four differential metabolites, including 3-hydroxybutyrate, acetate, glutamine, and glycine and a decrease in the serum of nine differential metabolites, including alanine, succinate, citrate, creatinine, o-phosphocholine, glucose, myo-inositol, tyrosine, and histidine compared with group A. Group B cows with IO had decreased glucose metabolism and impaired tricarboxylic acid cycle, increased lipid mobilization, and abnormal amino acid metabolism. The study provides a potential prevention strategy for IO in dairy cows in future.

Protective Effect of Xinmailong Injection on Rats With Myocardial Infarction.

This study aimed to investigate the protective effect of Xinmailong injection on rats with myocardial infarction. Thirty-six rats were induced myocardial infarction by operation, and six underwent sham operation. The myocardial infarction rats were randomly divided into three groups, 12 in each, and administered intraperitoneal injection of Xinmailong 5 mg/(kg·d), sodium creatine phosphate 80 mg/(kg·d), or normal saline as control respectively for 14 days. When the treatments were completed, the hemodynamic parameters of the rats were observed, and blood samples were taken to examine blood routine, blood coagulation index, liver and kidney function, inflammatory index, myocardial marker, thrombo-elastography, and other indicators. The morphology of cardiomyocytes was observed through light microscopy, and the microstructure of the myocardial cells was observed under electron microscope. No significant difference was found in blood routine, liver and kidney function, and blood coagulation index between the Xinmailong and sodium creatine phosphate groups compared with the saline control group. However, the inflammatory index and levels of myocardial markers were significantly decreased, and cardiac function was significantly improved. In terms of the morphology of myocardial cells, the Xinmailong group was similar to the sodium creatine phosphate group, the myocardial cell membrane was protected, and myocardial cell damage was reduced. In conclusion, Xinmailong is safe and had anti-inflammatory, heart-improving, and myocardial-protective effects. Its effectiveness is not inferior to that of sodium creatine phosphate.