Creatine Phosphokinase Activity Research Articles

Influence of 19-Nortestosterone on the Content of Steroid Hormones, Hematological Indicators and Individual Links of Metabolism in Tissues of Carp (Cyprinus carpio)

The influence of different concentrations of the synthetic steroid 19-nortestosterone on the body mass and indexes of internal organs, hematological parameters, testosterone, progesterone and cortisol content, certain links of metabolic processes and enzymatic activity in the tissues of carp (<i>Cyprinus carpio</i> L.) was studied. Under the short-term impact of 50 μg/dm<sup>3</sup>, 19-nortestosterone increased the total protein and albumins content, creatine phosphokinase and aspartate aminotransferase activity, decreased the inorganic phosphorus and iron content, alkaline phosphatase and alanine aminotransferase activity in the blood plasma and did not affect the respiratory rate, external body features, the mass of hepatopancreas and spleen and their indexes, hematological indicators, the steroid hormones concentration in blood plasma and hepatopancreas, content of glucose, triglycerides, creatinine, calcium, inorganic phosphorus in blood plasma. Under 200 μg/dm<sup>3</sup>, the respiratory rate and ESR increased, as well as the cortisol and testosterone content in blood plasma and hepatopancreas, whereas progesterone in blood plasma decreased and in hepatopancreas increased. High concentration of 19-nortestosterone caused elevated protein and albumin content, creatine phosphokinase and aspartate aminotransferase in the blood plasma activity, whereas glucose, inorganic phosphorus, iron, alkaline phosphatase activity was lower. The 19-nortestosterone did not affect the mass and index of hepatopancreas and spleen, hemoglobin concentration, the erythrocytes, leukocytes and lymphocytes number and ratio of eosinophils, neutrophils, lymphocytes and monocytes, as well as triglycerides, creatinine, calcium content, lactate dehydrogenase and alanine aminotransferase activity in blood plasma. The conclusion was made about the carp's ability to adapt to low concentrations of the synthetic steroid 19-nortestosterone under its short-term impact by modification of the certain links of metabolism and enzymatic activity. High concentration of 19-nortestosterone significantly affected the hormonal status and metabolism of carp.

Pathophysiological significance of creatinekinase and lactatedehydrogenase in the mechanisms of adaptation of muscle tissue of descendants

Over the past half century, the human population has seen a significant increase in the proportion of individuals exposed to ionizing radiation. One of the urgent problems of radiobiology is the establishment of the natural radiosensitivity of the organism, which provides the possibility of predicting the remote consequences of radiation exposure. The general condition of animals after ionizing radiation and the changes caused by this radiation largely determine the functioning of muscle tissue, which plays an important role in ensuring the vital activity of the body. The leading pathogenetic factor of radiation damage to the body is a violation of the metabolism and structural organization of enzymatic systems, due to which a sequence of biochemical processes and compartmentalization of enzyme systems change, and their solubilization increases. Additionally, it is extremely important to understand the detailed mechanisms of energy supply for the functioning of muscle tissue in the body of intact animals. The purpose of the work is to determine the pathophysiological significance of creatinekinase and lactatedehydrogenase in the mechanisms of adaptation of the body of the descendants of intact rats to the influence of a stress factor by studying changes in the activity of creatinekinase and lactatedehydrogenase in the blood and muscle system. A characteristic feature of creatine metabolism in 1-month-old pup rats was shown to be the low content of creatine and creatinine in skeletal and cardiac muscles against the background of low activity of the creatine phosphokinase enzyme in these muscles and its sharp increase in blood. Expressed hypercreatinuria and hypocreatininuria are registered in the blood of 1-month-old pup rats, which can be realized due to the strengthening of biosynthetic processes against the background of the inability of muscle tissue to fix these metabolites. In the skeletal and cardiac muscles of intact 1-month-old pup rats, the activity of creatine phosphokinase is decreased due to the decrease in the activity of the MM -isoform of the enzyme. The investigated metabolic processes in the muscle system of intact animals were shown to be extremely sensitive to the altering influence of ionizing radiation. The authors stressed that understanding the pathophysiological mechanisms of adaptation of the muscular system to the probable influence of a stressful radiation factor, we can draw a conclusion about the depth and severity of the pathological process based on the state of the enzyme systems, and we consider the determination of enzymatic tests in the blood to be important in diagnostic and prognostic aspects.

THE EFFECT OF DRY WHITE MISTLETOE EXTRACT ON THE CHANGE IN THE RATIO OF MITOCHONDRIAL SUBPOPULATIONS IN THE MYOCARDIUM IN ANIMALS WITH ALCOHOLIC CARDIOMYOPATHY

The aim of the research was to study the cardioprotective effect of dry white mistletoe extract in the context of assessing its effect on changes in the activity of interfibrillary and subsarcolemmal mitochondrial subpopulations of the myocardium in experimental alcoholic cardiomyopathy. Materials and methods. Alcoholic cardiomyopathy was modeled in female Wistar rats by course administration of ethanol at the rate of 3 g of absolute ethanol per 1 kg of animal body weight. The studied extract of white mistletoe leaves and the comparison drug – Trimetazidine – were administered orally at doses of 100 mg /kg and 35 mg/kg, respectively. Changes in troponin I concentration and serum creatine phosphokinase activity, citrate synthase activity in mitochondrial subpopulations, and the content of apoptosis markers – apoptosis-inducing factor and caspase 3 in myocardial tissue homogenate were determined during the work. Results and their discussion. The analysis of the results revealed that in rats with alcoholic cardiomyopathy, but without treatment, there is an increase in the concentration of troponin I and creatine phosphokinase activity, accompanied by an increase in the intensity of apoptosis reactions and citrate synthase activity of interfibrillary mitochondria. The use of Trimetazidine and the extract under analysis contributed to a decrease in troponin I content by 19.1% (p < 0.05) and 24.4%, respectively, and creatine phosphokinase activity by 19.8% (p < 0.05) and 28.4% (p < 0.05). It was also established that the activity of subsarcolemmal mitochondria in animals treated with Trimetazidine and white mistletoe extract increased by 121.9% (p < 0.05) and 306.3% (p < 0.05) with a decrease in the activity of citrate synthase of interfibrillary mitochondria by 27.6% (p < 0.05) and 41.4% (p < 0.05). It should be noted that administration of the comparative drug and the studied extract to rats resulted in a decrease in cardiomyocytes' apoptosis, expressed in a decrease in the concentration of apoptosis-inducing factor and caspase 3. Conclusions. In conditions of alcoholic cardiomyopathy, the negative interfibrillary phenotype of mitochondria prevails, which can contribute to the development of energy deficiency in the heart muscle, increasing the degree of myocardial damage. Based on the data obtained, it can be assumed that the extract of white mistletoe contains an active substance of cardioprotective activity associated with restoring the energy metabolism and suppression of apoptosis in the myocardium.

Evaluating Silymarin Extract as a Potent Antioxidant Supplement in Diazinon-Exposed Rainbow Trout: Oxidative Stress and Biochemical Parameter Analysis.

This study aimed to investigate the effects of diazinon on fish, focusing on hepatotoxic biomarkers and the potential protective effects of silymarin supplementation. One hundred eighty rainbow trout were randomly assigned to four groups: control, diazinon exposed (0.1 mg L-1), silymarin supplemented (400 mg kg-1), and diazinon + silymarin. Blood samples and liver tissue were collected after 7, 14, and 21 days of exposure to analyze biochemical parameters and oxidative biomarkers. Diazinon exposure in fish resulted in liver damage, as indicated by increased antioxidant enzyme activities in the hepatocytes. Silymarin showed the potential to mitigate this damage by reducing oxidative stress and restoring enzyme activities. Nevertheless, diazinon increased creatine phosphokinase activity, which may not be normalized by silymarin. Exposure to diazinon increased glucose, triglyceride, and cholesterol levels, whereas total protein, albumin, and globulin levels were significantly decreased in fish. However, silymarin controlled and maintained these levels within the normal range. Diazinon increased creatinine, urea, uric acid, and ammonia contents. Silymarin could regulate creatinine, urea, and uric acid levels while having limited effectiveness on ammonia excretion. Furthermore, diazinon increased malondialdehyde in hepatocytes, whereas administration of silymarin could restore normal malondialdehyde levels. Overall, silymarin showed potential as a therapeutic treatment for mitigating oxidative damage induced by diazinon in fish, but its effectiveness on creatine phosphokinase, glutathione reductase, and ammonia may be limited.

Candidate Genes for Prediction of Efficacy and Safety of Statin Therapy in the Kazakh Population.

The purpose of this research was to determine the frequency of mutation of the cytochrome CYP3A5 genes and transport proteins SLCO1B1 and MDR1 in patients with coronary heart disease in the Kazakh nation. A prospective cohort clinical and genetic study was conducted. The study was conducted in 2017-2019. Medical records containing information about drug prescription conducted in hospitals and outpatient departments were carefully analyzed. In the examined group of 178 patients treated with statins, a significant frequency of genetic variants that determine the increased risk of complications of statin use was revealed. There was a tendency toward an increase in the activity of creatine phosphokinase (CPK) in the blood upon detection of the A6986G mutation of the cytochrome gene and SLCO1B1 (c.521T>C) gene of the transport protein OATP1B1. In the studied Kazakh population, the presence of a homozygous mutant SLCO1B1 gene of the transport protein can be recommended as a genetic marker for the undesirability of using antihypercholesterolemic therapy with statins, which simultaneously leads to a decrease in the effectiveness of treatment and an increase in the risk of side effects.

Effect of testosterone in young ice-hockey players on hematological, biochemical parameters and the level of physical performance

Ice-hockey combines intense physical activity at high speed with elements of power struggle. Testosterone is the main anabolic hormone, an increase in which during adolescence is associated with faster growth, increased muscle mass, physical strength, and increased overall and aerobic performance. Thus, it is interesting to study the effect of different testosterone levels on metabolic parameters and physical performance of young ice-hockey players.Objective: to compare hematological and biochemical parameters, as well as exercise performance in young ice-hockey players, depending on the testosterone level.Materials and methods: the study included 100 young ice-hockey players aged 15–17 years (average age 15.3 ± 1.1 years). The study group included 25 young ice-hockey players with an increased level of total blood testosterone (> 27.5 nmol/l). The control group included 75 young athletes with normal testosterone levels. All athletes included in the study underwent the PWC 170 test to assess physical performance. Assessment of hematological parameters included hemoglobin (HGB), hematocrit, red blood cell, mean corpuscular volume (MCV), mean concentration hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC). β-crosslaps, myoglobin and creatine phosphokinase (CPK) activity were measured to assess the functional state of muscle and bone tissue.Results: A comparative assessment of the CBC in young ice-hockey players did not reveal significant differences depending on the level of serum testosterone level. Tolerance to physical activity in the study groups also did not differ. Young ice-hockey players with increased testosterone levels showed lower β-crosslaps values compared to the control group. The values of other biomarkers of the functional activity of muscle tissue did not depend on the level of testosterone levels.Conclusion: Increased serum testosterone level in the range from 27.5 to 40 nmol/l, detected in young ice-hockey players aged 15–17 years, is not associated with a change in hematological and biochemical parameters. High total testosterone levels do not improve physical performance. It is necessary to continue studies to assess the dynamics of the serum testosterone in young ice-hockey players to clarify the long-term effects of androgens on the metabolic and functional indicators of young athletes. Increasing serum testosterone levels in athletes is subject to wide discussion in the community of sports physicians and endocrinologists.

Analysis of the Serie of Cases of Neuroleptic Malignant Syndrome (Cross-Sectional Observational Study)

Background: neuroleptic malignant syndrome (NMS) is the most dangerous complication of therapy with neuroleptics with high mortality. The publications on MNS are the reviews articles or case reports.The aim of study: an identification of the frequency of occurrence of NMS harm in patients of the city psychiatric hospital over a 4-year period, a clarification of the clinical manifestations of this complication, an analysis of the effectiveness of the treatment and rate of deaths.Patients and methods: 19 cases with a confirmed diagnosis of NMS who were treated for 4 years in one of the clinical psychiatric hospitals in Moscow with 1158 beds. All patients met the criteria for the diagnosis of NMS according to DSM-5 (333.92). The severity of extrapyramidal disorders was assessed on the Simpson–Angus Rating Scale (SAS). The Bush–Francis Catatonia Rating Scale (BFCRS) was used to assess the features and severity of catatonic symptoms. All patients underwent a study of the general clinical analysis and biochemical analysis of blood with the determination of the number of leukocytes and the activity of the enzyme creatine phosphokinase (CPK).Results: the incidence of NMS was 0.035% of all treated patients, mortality— 10.5%. Most often, NMS develops during treatment with haloperidol and much less frequently with the appointment of atypical antipsychotics risperidone and clozapine in patients with schizophrenia, acute polymorphic mental disorder, organic mental disorders, including dementia. The influence of such factors as gender, age, type of disease on the development of NMS has not been established. The age of patients with a fatal outcome was higher than the average for the group. The clinical picture of NMS was characterized by a combination of catatonic, extrapyramidal and somato-vegetative disorders with hyperthermia, leukocytosis and increased activity of CPK.Conclusion: today NMS remains a relatively rare, but extremely dangerous complication of antipsychotic therapy, with a high risk of death and can develop both in the treatment with first and second generation antipsychotics in patients with schizophrenia and acute polymorphic mental disorder and in organic mental disorders, including dementia.

Biochemical parameters of blood and tissue of the gastrocnemius muscle in chronically alcoholized rats under oral administration of C(60) fullerene aqueous solution

Biochemical indices of blood and tissue of the gastrocnemius muscle chronically alcoholized (for 3, 6 and 9 months) rats were studied. С60 fullerene aqueous solution (C60FAS) was administered orally as a pharmacological agent at a dose of 1 mg/kg daily throughout the experiment in a three routes: 1 h before alcohol intake (preventive regimen), together with alcohol (therapeutic regimen I) and 1 h after alcohol intake (therapeutic regimen II). Creatine phosphokinase (CPK), lactate dehydrogenase (LDH), catalase, superoxide dismutase, glutathione peroxidase (GPx) activity and the level of creatinine, lactate, hydrogen peroxide, reduced glutathione were estimated with clinical diagnostic kits. A pronounced upward trend in creatinine and lactate content, CPK and LDH activity with increasing degree of alcoholic myopathy during experiment was detected. Administration of C60FAS was shown to reduce the biochemical indices of muscle injury and to reduce oxidative processes by maintaining the balance between pro-oxidant and antioxidant systems. The maximum positive effect was observed when C60FAS was administered together with alcohol (therapeutic regimen I). The results indicate on C60 fullerene ability to correct the pathological condition of the muscular system arising from alcohol intoxication. Keywords: alcohol intoxication, antioxidant system, C60 fullerene, creatine phosphokinase, gastrocnemius muscle, lactate dehydrogenase

Biochemical indicators of sperm plasma and morphofunctional features of spermatozoa in HIV-infected men

BACKGROUND: The problem of male infertility particularly affects men with HIV infections, as semen volume and sperm motility are reduced by highly active antiretroviral therapy.
 AIM: To analyze the metabolic parameters of spermatozoa, spermograms and spermatozoa morphology in HIV-infected men.
 MATERIALS AND METODS: The study included 47 patients (aged 2546 years) who were under observation at the clinic of Professor M.A. Florova (Samara). 2 groups were formed: main (n=22) HIV-infected patients who want to have children (with an undetectable viral load (4050 copies/ml) and taking antiretroviral therapy); control (n=25) clinically healthy men with one child. The material was taken and the ejaculate was examined according to standardized methods proposed by WHO experts.
 RESULTS: In HIV-infected men, spermatozoa motility was low (the number of progressively motile spermatozoa was 28.503.72%), the spermatozoa concentration was two times lower compared to the control group of men, the morphological characteristics of spermatozoa were significantly worse than in the control group: most often pathology of the head (21.751.10%) and neck of the spermatozoon (22.301.18%) was detected, which negatively affects the fertilizing ability.
 In HIV-infected men, changes in metabolic metabolism were noted: the activity of creatine phosphokinase in both sperm plasma (661.951.08 U/l) and blood plasma (76.901.09 U/l) was lower, than in the control group (844.250.13 and 79.501.37 U/l, respectively), the glucose concentration (8.071.14 mmol/l) was 2 times higher than in the control group (3.061.09 mmol/l), the concentration of calcium 6.530.01 and sodium (119.201.23 mol/l) slightly exceeded those in the group of healthy patients (5.550.08 and 116.850.01 mol/l, respectively). Electron microscopic analysis revealed fragmentation of sperm DNA: the highest percentage of sperm with fragmented DNA was found in men with HIV infection (more than 23%).
 CONCLUSION: It was revealed that in vivo fertilization in HIV-infected men is impossible in most cases. The study forms the basis for a future comprehensive assessment of the state of reproductive function in HIV-infected men to assess their fertility and the need for assisted reproductive technologies.

Effect of Graded Levels of Dried Orange (Citrus sinensis) Byproducts on Production Efficiency, Blood Parameters and Antioxidant Status of Broiler Chickens

Background: Orange wastes are bioactive compounds produced during the industrial transformation.The experiment was conducted to evaluate the effects of feeding dried sweet orange byproducts (DOP) on broiler growth performance, serum metabolites and antioxidant status. Methods: A total of 200 of 20-day experimental broiler chicks were distributed into a completely randomized design which included 4 dietary treatments with 5 replicates of 10 birds fed each. The dietary treatments included a control diet with no corn substitution and three diets containing 5, 10 and 15 % Dried Orange Pulp. Feed intake (FI), body weight gain (BWG), feed conversion ratio (FCR), carcass traits, serum components, and blood antioxidant status were measured. Result: FCR and BWG of chickens were improved when using graded levels of DOP compared to the control group (P less than 0.05). Serum total protein and glucose were significantly increased (P less than 0.0001) in DOP groups compared to the control. Serum Cholesterol and triglyceride decreased significantly (P less than 0.0001) in the DOP groups. Higher Aspartate amino transferase, Lactate Dehydrogenase and Creatine Phospho Kinase activities were observed in Control group compared to the other treatments.

Влияние гексанового экстракта Serenoa repens на функцию мочевого пузыря при частичной инфравезикальной обструкции.

Introduction. It is believed that the «target» of the hexane extract of Serenoa repens (HESR) is the prostate gland itself. However, there is evidence in the literature that the therapeutic effect of HESR can be carried out at the level of the bladder. We hypothesized about the direct effect of HESR on the obstructive bladder, preventing the development of its dysfunction. Material and methods. The study was conducted on 30 white mongrel female rats. Bladder outlet obstruction (BOO) was modeled by applying a ligature to the urethra on 18G catheter. The animals were divided into groups: experimental – HESR (n = 10); sham/placebo (n = 15) and control (n = 5). From the day following the operation, rats of the experimental series daily received aqueous alcohol suspension of HESR (27.6 mg/kg) intragastrically through a probe, and the sham group received the same volume of 3.5% alcohol solution. After 2 weeks, the survival rate of the animals, the functional state of the bladder, daily diuresis, biochemical changes in blood and urine characterizing the functional state of the kidneys were evaluated. Bladder function was assessed according to infusion cystometry. After examination, the animals were euthanized and the bladders was removed to determine its mass and histological examination. Results. Within 2 weeks in the sham group 10 (67%) rats died due to renal failure, while in the HESR group – only 3 (30%). An obvious overfilling of the bladder in the main and sham groups was revealed. Bladder volume was larger in the HESR group compared to the sham (p<0.05) one. The daily diuresis of the animals of the main group was also higher compared to the sham group, and the decrease in diuresis in the sham group was lower compared to the control (p=0.045). The functional state of the bladder was assessed by filling cystometry. In the sham group, an increase in intravesical pressure occurred immediately after the start of fluid infusion and its growth was accompanied by irregular fluctuations, which is a manifestation of secondary detrusor hyperactivity, while in the HESR group, a gradual increase in pressure without fluctuations up to large volumes of fluid was observed. The dynamics of the volume/pressure index in the main group was close to normal, which indicates better compliance of the wall of the hypertrophied bladder during HESR therapy. The use of HESR contributed to better preservation of kidney function. In terms of daily diuresis, glomerular filtration rate, tubular reabsorption of sodium, daily excretion of urea and creatinine in the urine, statistically significant differences were established between the main and sham groups. In the sham group, a more pronounced activity of creatine phosphokinase was noted compared to the HESR group (p<0.05), which indicates a significant damage to kidney and bladder cells (cytolysis) during BOO. Animals of both groups showed pronounced detrusor hypertrophy. If in the sham group the mass of the bladder exceeded the average value of the control group by 5.7 times, then in the HESR group it was 10.5 times. Histological studies confirmed the development of bladder hypertrophy, which was statistically significantly more pronounced in the HESR group: an increase in the thickness of the detrusor (p<0.001) and bladder wall (p<0.05). At the same time, the area of perimuscular sclerosis foci was significantly larger in the sham group (p<0.05), which indicates bladder decompensation. Thickening of the bladder wall also occurred due to the submucosal layer, which was more noticeable in the sham series (p<0.05) and was associated with the presence of edema, inflammatory infiltrates, and sclerosis in this area. In half of the cases of the sham series, de-epithelialized areas of the bladder mucosa were observed, while in the HESR group they were absent. In the experimental series, in all samples, a pronounced expansion of the vessels of the submucosal layer was noted. At the same time, if in the experimental group inflammatory infiltration was detected only within the submucosal layer, then in half of the cases of the sham series, diffuse infiltration of the entire bladder wall was detected. Conclusion. In BOO detrusor hypertrophy, the use of HESR contributes to the better preservation of the functional parameters of the bladder (compliance and evacuation ability), reduces the degree of inflammation and sclerosis of the bladder wall, prevents the development of decompensation of the bladder and, thereby, contributes to the preservation of kidney function. A significant decrease in the frequency of development and the degree of detrusor hyperactivity against HESR makes it possible to think about it’s direct effect on the smooth muscle cells of the bladder wall and/or on the way of regulating their contractile activity. The mechanisms of the HESR effects on the BOO bladder are currently unclear. This is probably of a complex nature, where the anti-inflammatory effects of Serenoa Repens are expected to play a primary role.

Downer cows: a reanalysis of an old data set

ABSTRACT Aims To compare the performance of two predictive models for the survival of downer cows. Methods The first model had been developed in 1987 using a dataset containing missing values, while the second, new model was developed on the same dataset but using modern data imputation and analytical methods. Missing data were imputed using multiple imputation by chained equations and a logistic regression model fitted to the imputed data, with survival or not as the outcome variable. The predictive ability of the model built on the imputed data was contrasted with the original prognostic model by testing them both on a second smaller but complete data set, collected contemporaneously with the development of the original model but from a different region of New Zealand. Sensitivity, specificity, accuracy, and cut point for the two models were calculated. Results The original 1987 model had a slightly higher accuracy than that of the new one with a sensitivity of 0.85 (95% CI = 0.72–0.94) and a specificity of 0.82 (95% CI = 0.7–0.91), using a cut point for the probability of survival = 0.313. Conclusions The original prognostic formula published by Clark et al. in 1987 performed as well as a modern model built on an imputed data set. Clinical relevance The use of a prognostic test based on the Clark model should remain an important part of the clinical examination of downer cows by New Zealand veterinarians. Abbreviations: AUC: Area under the curve; AST: Aspartate transaminase activity; CK: Creatine phosphokinase activity; GAM: Generalised additive model; NSAID: Non-steroidal-anti-inflammatory drugs; PCV: Packed cell volume

Anti-fatigue activity of dripped spent hens chicken essence in ICR mice.

Chicken essence and branched chain amino acid (BCAA) supplementation has been recognized to significantly relieve fatigue. To obtain chicken essence with high amounts of BCAA, spent hens herein was used to prepare dripped chicken essence (SCE) and compared with commercial dripped chicken essence (CCE) for in vivo anti-fatigue effect. To determine the effect on anti-fatigue by dripped chicken essence, the exhaustive swimming was performed. Thirty-two 7-week ICR mice were divided into four groups, which included the control group (CG), CCE, SCE-1X and SCE-2X. The mice were given daily oral administration (0.012 mL/g body weight/d). The fatigue index analysis was conducted weekly. The results showed that SCE had a higher BCAA level as expected, and mice treated with dripped chicken essence (CCE and SCE) could significantly improve exercise performance. The lower blood lactate level, blood urea nitrogen level and creatine phosphokinase activity were found in the supplement of SCE group compared with the CCE group, which suggested that the SCE possessed strong anti-fatigue ability. This could possibly be due to the higher content of BCAA. In this study, SCE promoted recovery from physical fatigue in mice and elevated endurance ability. Among them, the double dose (SCE-2X) showed the strongest anti-fatigue ability. Taken together, spent chickens could be a good source of chicken essence to improve the effect of anti-fatigue.

ENZYME ACTIVITY OF CARBOHYDRATE AND AMINOACID METABOLISM, LIPID PEROXIDATION IN CARP TISSUES UNDER THE EFFECT OF 19-NORTESTOSTERONE

The aim of the study was to investigate the activity of a number of enzymes of carbohydrate metabolism, individual amino acids, creatine phosphate, inorganic phosphorus and lipid peroxidation (LPO) in the hepatopancreas and blood plasma of carp (Cyprinus carpio L.) under the influence of the synthetic steroid 19-norethisterone. The experiments were conducted on two-year-old carps, which were kept for 24 hours in aquariums with different concentrations of 19-nortestosterone in water at optimal temperature, oxygen content and pH. The activity of lactate dehydrogenase (LDH), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine phosphokinase (CPK) in the supernatant fraction of hepatopancreatic homogenate was monitored by UV-kinetic and colourimetric methods. The activity of isocitrate dehydrogenase (ICDH) was determined spectrophotometrically by the accumulation of NADPH, and lactate dehydrogenase (LDH), glucose-6-phosphate dehydrogenase (Gl-6-PDH) and glucose-6-phosphatase (Gl-6-Phase) - by the accumulation of NADH in the reactions of conversion of the corresponding substrates. Conventional biochemical methods were used to determine the concentration of diene conjugates, malondialdehyde (MDA), lipid hydroperoxides, catalase (CAT) and superoxide dismutase (SOD) activity in blood plasma and supernatant fraction of hepatopancreatic homogenate. Conventional biochemical methods were used to determine the concentration of diene conjugates, malondialdehyde (MDA), lipid hydroperoxides, catalase (CAT) and superoxide dismutase (SOD) activity in blood plasma and the supernatant fraction of hepatopancreatic homogenate. It was found that fish exposed to water with a concentration of synthetic steroid 19-nortestosterone of 50 µg/dm3 for 24 hours had higher levels of MDA and diene conjugates, increased activity of CAT and SOD in blood plasma, increased activity of ALP, ALT, AST, CK, CAT and SOD in the hepatopancreatic supernatant fraction, and LDH remained unchanged, as well as the level of lipid hydroperoxides in blood plasma. Under the influence of 19-nortestosterone, the activity of Gl-6-FDG and IDDG in the mitochondrial fraction of carp hepatopancreas increases, and that of Gl-6-phase in the supernatant. At a concentration of 19-nortestosterone in water of 200 μg/dm3, the content of lipid hydroperoxides in the blood plasma of carp increases, and in the hepatopancreatic supernatant fraction - diene conjugates and SOD activity, while ALP and CPK decrease relative to their values in fish with a low level of xenobiotic in water. The content of MDA, diene conjugates, as well as the activity of CAT, SOD, ALP, CPK and LDH in blood plasma, IDH and Gl-6-PDH in the mitochondrial fraction and Gl-6-Phase and LDH in the hepatopancreatic supernatant did not change, but remained higher than in the control. It was concluded that 19-nortestosterone stimulates carbohydrate metabolism, amino acids alanine and aspartate transamination, creatine phosphate conversion, antioxidant enzyme activity and lipid peroxidation in the hepatopancreas of two-year-old carp. The obtained results deepen the understanding of the mechanism of action of synthetic steroids in bony fish, and the differences in enzyme activity and lipid peroxidation can be important criteria for ecotoxicological assessment of water bodies contaminated with hormonal compounds.

Adult spinal muscular atrophy: problems of early diagnosis

Background. Spinal muscular atrophy (SMA) associated with chromosome 5q is an autosomal recessive neuromuscular disease caused by a mutation in the SMN1 gene. Adult forms of SMA are relatively rarer and associated with significant diagnostic difficulties and pronounced delay in diagnosis. The available pathogenetic therapy for SMA has the greatest effect with an earlier start of treatment, so timely diagnosis of the disease significantly improves the overall prognosis.Aim. To evaluate the features of diagnosis of the adult SMA and summarize the first experience of such diagnosis in Russian patients.Materials and methods. We analyzed the archived medical records of 50 adult patients with SMA consulted at the Research Center of Neurology (Moscow).Results. The data of patients with SMA type 2 (n = 26), SMA type 3 (n = 21), SMA type 4 (n = 3) were analyzed. The delay time for diagnosis in SMA type 2 is 35 (0–270) months, with SMA types 3 and 4 – 108 (0–408) months. The diagnosis of SMA was the first diagnosis in SMA type 2 in 69 % of cases, in SMA types 3 and 4 in 29 % of cases. The most common first diagnosis in patients with SMA is myopathy, accounting for 52 % of all misdiagnosed cases. A small percentage of the use of needle electromyography in the diagnostic process was noted (1/3 of cases); in cases of its use, this did not accelerate the correct diagnosis. Creatine phosphokinase activity is often elevated in patients with SMA types 3 and 4 compared with SMA type 2 (p <0.05). Conclusions. To reduce the delay in the correct diagnosis of SMA and earlier initiation of pathogenetic therapy, it is necessary to increase the awareness of specialists about the features of diagnosis the disease and focus on alternative erroneous diagnoses, among which adult patients with SMA may “hide”. The key to confirming the diagnosis is DNA testing.

CMV-associated carditis in a 5-month-old child

The article presents a case of cytomegalovirus-associated carditis in a 5-month-old child. Carditis proceeded in an arrhythmic form and was manifested by frequent ventricular extrasystoles. An increase in the activity of creatine-phosphokinase was noted in the blood. The isolation of cytomegalovirus DNA from the blood, as well as IgM and IgG to cytomegalovirus, made it possible to associate the development of carditis with IgG to cytomegalovirus. Carditis in a child proceeded without signs of heart failure. After diagnostic and therapeutic procedures, the child was discharged home. This case shows that one of the possible causes of carditis in children is cytomegalovirus, which necessitates testing for markers of active cytomegalovirus infection. In this age group, it is possible to register both congenital and acquired variants of infection with heart damage, having different treatment regimens and prognosis.

Use of branched-chain amino acids for reducing exercise-caused skeletal muscle damage

Introduction: Skeletal muscles damage (direct and vicarious) slows down the recovery processes in patients with injuries of the musculoskeletal system. It occurs in the early postoperative period as well. An increase in the rigidity of the skeletal muscle extracellular matrix can reduce pain, tissue swelling, and accelerate the recovery of contractility.Objective: The analyses of the effect of branched-chain amino acids (BCAAs) intake on the expression of IGF1 genes, type 1, 3 and 5 collagen, which are crucial in the composition of the skeletal muscle extracellular matrix, as well as on the muscle membrane damage against the background of chronic damage to skeletal muscles.Material and methods: 12 young healthy male subjects, skiers aged 19 (18; 22) received a placebo treatment (maltodextrin, 100 mg/kg body weight/day; n = 6) or a mixture of amino acids (leucine, isoleucine, valine – 50:25:20 mg/kg body weight/day respectively; n = 6). The treatment was received daily against the background of a large amount of aerobic high-intensity training (up to 22 hours per week). Before and after the amino acids intake a biopsy of the musculus vastus lateralis was performed, and venous blood samples were taken during the experiment.Results: The intake of leucine against the background of training led not only to a pronounced increase in the level of IGF1 protein in blood by 1.5 times (which corresponds to the literature data), but also to a trend towards an increase in the expression of IGF1Ea mRNA by 1.8 times in the skeletal muscle, and a decrease in the level of markers of muscle membranes damage – creatine phosphokinase (CPK) activity and myoglobin. In addition, changes in the IGF1-dependent collagen genes expression strongly correlated with changes in IGF1Ea expression, but not with IGF1 protein in blood (pooled group, n = 12). Thus, the intake of leucine as a part of the essential amino acids can reduce damage to skeletal muscles caused by excessive physical activity, lack of physical activity, or direct trauma.Conclusion: A 10-week BCAAs intake by individuals with documented chronic muscle membrane damage caused an increase of basal levels of IGF1 in blood and a trend towards increased IGF1Ea mRNA expression in skeletal muscle, and also caused a modest reduction in damage of skeletal muscle membrane.

Ischemic preconditioning/ischemic postconditioning alleviates anoxia/reoxygenation injury via the Notch1/Hes1/VDAC1 axis.

Ischemic preconditioning (IPC), and ischemic postconditioning (IPost) have a significant protective effect on myocardial ischemia/reperfusion (MI/R) injury by alleviating oxidative stress and mitochondrial disturbances, although the underlying molecular mechanisms are unclear. The study was to demonstrate that cardioprotection against anoxia/reoxygenation (A/R) injury is transduced via the Notch1/Hes1/VDAC1 signaling pathway. Using mass spectrometry and tandem affinity purification (TAP), to screen for differentially expressed proteins associated with Hes1, followed by standard bioinformatics analysis. The co-immunoprecipitation (Co-IP) assay confirmed an interaction between Hes1 and VDAC1 proteins. H9c2 cells were transfected with Hes1 adenoviral N-terminal TAP vector (AD-NTAP/Hes1) and Hes1-short hairpin RNA adenoviral vector (AD-Hes1-shRNA) to establish A/R injury, IPC, and IPost models, respectively. The expression of Hes1 and VDAC1 proteins were measured by western blot analysis, while the levels of reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm), and apoptosis were evaluated by flow cytometry. AD-NTAP/Hes1 can activate the exogenous protein expression of Hes1, thus decreasing creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) activity and promoting cell viability. The study found that VDAC1 was a potential target protein for Hes1 and the overexpression of Hes1 protein expression downregulated protein expression levels of VDAC1, reduced ROS production, stabilized ΔΨm, and inhibited apoptosis in H9c2 cells. Additionally, downregulation of Hes1 protein expression also upregulated VDAC1 protein expression, increased ROS production, imbalanced ΔΨm, promoted cell apoptosis, and attenuated the cardioprotection afforded by IPC and IPost. The Notch1/Hes1 signaling pathway activated by IPC/IPost can directly downregulate the protein expression of VDAC1 and consequently relieve A/R injury.

Toxic effects of polyethylene microplastics on transcriptional changes, biochemical response, and oxidative stress in common carp (Cyprinus carpio)

Aquatic ecosystems have become a place for accumulating microplastics (MPs). MPs can directly or indirectly damage organisms. Although studies of the toxicity of MPs, there are insufficient literature reports on the effects of MPs on freshwater aquatic life. Therefore, this study aimed to evaluate the effect of MPs toxicity on Cyprinus carpio. In this study, biochemical parameters, oxidative biomarkers, and gene expression were assayed in fish exposed to 0, 175, 350, 700, and 1400 μg L−1 of MPs for 30 days. MPs were detected in the liver and intestine of fish using FTIR-analysis. Mt1, Ces2, and P450 mRNA expression were enhanced in the hepatocytes of fish exposed to MPs, while Mt2 gene expression was significantly decreased. After exposure to MPs, MDA and carbonyl protein levels were higher than those of the reference group. The antioxidant capacity and glycogen contents in the hepatocytes significantly declined. MPs significantly inhibited glutathione reductase (GR), glucose 6-phosphate dehydrogenase (G6PDH), and catalase (CAT) activities. However, superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities increased. MPs decreased the total protein, globulin levels, and butyrylcholinesterase (BChE) activity in blood. In contrast, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), and creatine phosphokinase (CPK) activities increased in treated-fish with MPs. Glucose, creatinine, cholesterol and triglyceride concentrations in fish exposed to MPs were significantly higher than that of the reference group. Consequently, MPs exposure could disrupt biochemical homeostasis, oxidative stress and alter the expression of genes involved in detoxification.

Differential diagnosis of myopathy and multiple epiphysal dysplasia caused by mutations in the <i>COMP</i> gene in children

Background. Multiple epiphysal dysplasia (MED) type 1 (OMIM: 132400) is one of 7 genetic variants of this group of skeletal dysplasias described to date. The disease is caused by mutations in the COMP gene located on chromosome 19p13.1. The presence of muscle hypotonia and ligamentous laxity, as well as a moderate increase in the level of creatinephosphokinase activity, can lead to misdiagnosis of myopathy.Objective: to analyze the clinical and genetic characteristics of type 1 MED caused by mutations in the COMP gene in a series of Russian patients. Differential diagnosis was focused on the distinctive features of the disorder and hereditary myopathies.Materials and methods. We observed 8 patients from 7 families aged 7 to 15 years with MED type 1 caused by heterozygous mutations in the COMP gene. To confirm the diagnosis, the following methods were used: genealogical analysis, clinical examination, neurological examination with psycho-emotional testing, radiography and targeted sequencing of a panel consisting of 166 genes responsible for the development of inherited skeletal pathology.Results. Case history, clinical, radiological and genetic characteristics of 8 patients with MED type 1 caused by mutations in the COMP gene were analyzed. The first clinical manifestations of the disease were recorded from the age of 2–3 years and were characterized by gait disturbances, muscle weakness, difficulties with climbing stairs, frequent falls when walking, the inability to get up from the floor and from a squatting position and hypermobility of the joints. Electroneuromyographic study did not reveal the signs of miopathy. In two patients, a moderate increase in the creatinekinase level of up to 250–360 u / l was found. All patients were surveyed by neurologists for several years with a clinical diagnosis of congenital myopathy. At the age of 5–6 years patients COMPlained knee and ankle pain, which was assumed as rheumatic arthropathy. X-ray examination revealed typical signs of deficient ossification of the epiphyses. The next-generation sequencing analysis revealed seven single nucleotide variants in the COMP gene that lead to MED type 1. Three of the found variants here identified for the first time. As previously described, the majority of nucleotide variants (six out of seven) were localized in the 8–14 exons of the COMP gene and led to amino acid substitutions in calmodulin-like protein domain repeats, and only one substitution was localized in the C-terminal region of the protein molecule.Conclusion. In most patients with MED caused by mutations in the COMP gene, the first symptoms of the disease are gait disturbance, muscle weakness, and Gowers» maneuvers. The presence of these symptoms, along with a moderate increase in the level of creatinephosphokinase activity, often precedes the onset of clinical manifestations of skeletal dysplasia, leading to a misdiagnosis with myopathies. Accession of expressive arthralgias to these symptoms was mistakenly identified as reactive arthritis. X-ray examination of patients’ long bones helps to suspect the presence of MED. This X-ray imaging shows specific signs of epiphyses damage. A molecular-genetic analysis needs to be done to diagnose the genetic variant, caused by mutations in gene COMP.