CPK Activity Research Articles

Comparative assessment of the most important markers of equine myopathy in connection with creatine kinase activity

Purpose: to assess the activity of AST, LDH and HBDH enzymes in relation to different levels of total creatine phosphokinase activity to understand the degree of their diagnostic value in determining the severity of myopathy in horses.Materials and methods. For the study, statistical processing of a data array of the results of a biochemical study of blood serum from 247 horses was carried out. Taking into account the reference intervals of CPK activity, all results were ranked by increasing creatine phosphokinase activity and divided into four groups. In each group, the average values for AST, LDH and HBDH were calculated, and the degree of significance of intergroup differences was calculated using the Student's t test. The correlation between the activity of the enzymes AST, LDH and HBDH with the activity of CPK in each group was also assessed.Results. An increase in the activity of creatine phosphokinase in horses by 59,4—409,6 % is accompanied by an increase in the activity of the enzymes AST by 14,5—77,2 %, LDH by 17,8—33,0 % and HBDH by 17,1—27,1 %. When comparing Pearson's correlation coefficients across the entire study sample, a significant direct relationship is determined between CPK and AST and a moderate direct relationship when comparing CPK with LDH and HBDH. Calculation of the Pearson coefficient in each of the studied groups showed different degrees of correlation between the creatine phosphokinase indicator and other enzymes, but in all cases positive. A study of the percentage contribution of HBDH to the total lactate dehydrogenase activity showed an inversely proportional relationship with respect to the activity of creatine phosphokinase, which indicates massive muscle damage with switching energy metabolism in them.Conclusion. The conducted studies made it possible to more deeply understand the cause-and-effect relationships of changes in the activity of enzymes that are specific to muscle tissue in connection with the severity of myopathy. The data obtained can assist veterinary specialists in analyzing the results of biochemical blood tests in horses.

A Calcium-Dependent Protein Kinase Regulates the Defense Response in Citrus sinensis.

Citrus Huanglongbing (HLB), which is caused by 'Candidatus Liberibacter asiaticus' (CLas), is one of the most destructive citrus diseases worldwide, and defense-related Citrus sinensis gene resources remain largely unexplored. Calcium signaling plays an important role in diverse biological processes. In plants, a few calcium-dependent protein kinases (CDPKs/CPKs) have been shown to contribute to defense against pathogenic microbes. The genome of C. sinensis encodes dozens of CPKs. In this study, the role of C. sinensis calcium-dependent protein kinases (CsCPKs) in C. sinensis defense was investigated. Silencing of CsCPK6 compromised the induction of defense-related genes in C. sinensis. Expression of a constitutively active form of CsCPK6 (CsCPK6CA) triggered the activation of defense-related genes in C. sinensis. Complementation of CsCPK6 rescued the defense-related gene induction in an Arabidopsis thaliana cpk4/11 mutant, indicating that CsCPK6 carries CPK activity and is capable of functioning as a CPK in Arabidopsis. Moreover, an effector derived from CLas inhibits defense induced by the expression of CsCPK6CA and autophosphorylation of CsCPK6, which suggests the involvement of CsCPK6 and calcium signaling in defense. These results support a positive role for CsCPK6 in C. sinensis defense against CLas, and the autoinhibitory regulation of CsCPK6 provides a potential genome-editing target for improving C. sinensis defense. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Effects of Fluoride on Metabolic enzymes of brain and Gastrocnemius muscle of mice: protective role of selenium and alpha-tocopherol

The present study aimed to investigate the ameliorative effects of Selenium and Alpha-tocopherol on activities of enzymes involved in metabolic functions, energy production and transfer in the brain and gastrocnemius muscle of mice treated with sodium fluoride. Animals were separated into five groups and treated with Selenium (05 µg/kg BW), Alpha-tocopherol (2mg/kg BW) alone, and in combination for 15 days to evaluate their ameliorative effects against fluoride (20mg/kg BW) toxicity. The obtained results showed a statistically significant (p<0.05) decrease in the body weight, organo-somatic index, protein content, ALAT, AAT, CPK, acid phosphatase and alkaline phosphatase enzymes activities in the fluoride-treated group in comparison with control group indicates oxidative stress induced by fluoride. The effect of fluoride on enzymes of muscle was comparatively higher, which suggests the greater accumulation of fluoride in muscle than brain. These changes were reverted moderately in Selenium and alpha-tocopherol alone treated groups, and significantly (p<0.05) in Selenium + alpha-tocopherol treated group suggests the combinatorial effect of Selenium and alpha-tocopherol in the amelioration of fluoride-induced toxicity. In conclusion, the findings all pointed to the fact that Selenium and alpha-tocopherol showed combinatorial effects in the amelioration of oxidative stress induced by fluoride.

Cardiotoxic Effects of Hemiscorpius Lepturus Scorpion Venom Fractions in Rats

Background: Scorpion stings are responsible for many deaths in humans; however, the toxicity mechanisms of the venoms from many species are not well studied. We investigated the cardiotoxicity of the crude venom from H. lepturus scorpion and its isolated fractions, F-I to F-VI. Methods: The scorpion’s venom was extracted into six fractions by chromotagraphy. Healthy male Wistar rats (N=72) were equally divided into eight groups of nine: G1: Controls (0.5ml. normal saline), G2: Crude venom (1000µg/kg), G3: F-I (120µg/kg), G4: F-II (430µg/kg), G5: F-III (80 µg/kg), G6: F-IV (180µg/kg), G7: F-V (60µg/kg), and G8: F-VI (130µg/kg). Blood samples were obtained by cardiac puncture at 1, 3 and 24 hours after the venom injection. The serum levels of AST, LDH, CPK, CK-MB and troponin-I were determined. Upon euthanasia, the hearts were removed from the rats and examined microscopically for histopathology. Results: In groups receiving crude venom and F-VI, we observed multifocal fragmentation of myocardial fibers, hemorrhage, degeneration and disappearance of striations in cardiac muscles as compared to the controls. The findings showed that AST and LDH activity in groups 2, 4 and 8, CPK activity in groups 2, 4, 6 and 8 and CK-MB activity and troponin-I levels in groups 2 and 8 increased significantly compared to those in the control group. Conclusion: There was evidence of significant cardiotoxicity in the group receiving crude venom and F-VI. Although alterations in the enzymatic and troponin-I levels were observed in other groups, the greatest cardiotoxicity of H. lepturus venom was caused by fraction VI.

Кардіопротекторна дія індукторів синтезу білка KLOTHO за умов гострого експериментального інфаркту міокарда

Останніми десятиліттями в світі відзначається тенденція до неухильно зростаючого розповсюдження судинних захворювань, що зумовлено загальним старінням населення, збільшенням захворюваності на цукровий діабет, артеріальну гіпертензію й атеросклероз. Сьогодні в лікуванні захворювань серцево-судинної системи гiпоксичного генезу широко використовуються препарати, що відновлюють кровоток, а також препарати, які впливають на метаболізм клітини, а саме, на окремі ланки адаптаційних молекулярно-біохімічних реакцій. Відносно останніх препаратів накопичено багато експериментальних і клінічних даних щодо їхньої ефективності. Однак з точки зору доказової медицини ефекти даних препаратів суперечливі та сумнівні. У зв’язку з цим йде активний пошук препаратів метаболітотропної дії.Мета дослідження – встановлення кардіопротекторної дії індукторів експресії та синтезу білків Klotho (вітамін D, Necrostatin-1) в умовах моделювання експериментального інфаркту міокарда.Експериментальну частину роботи виконано на 40 статевозрілих щурах – самцях масою 170–210 г. Гострий інфаркт міокарда (ГІМ) моделювали шляхом введення впродовж 4 діб пітуїтрину (1 Од/кг підшкірно) та адреноміметика ізопреналіну (200 мг/кг внутрішньом’язово). Досліджувані препарати – Вітамін D (5000 МО/доба) (Гмбх Арцнайміттель, Німеччина) вводили внутрішньошлунково і Necrostatin-1 (Nec-1 25 mg, Sigma-Aldrich) – 1 мг/кг внутрішньоочеревинно через 20 хв після введення ізадрину впродовж 4 днів. Наявність ГІМ підтверджували шляхом визначення його маркерів у плазмі крові імунохемілюмінесцентним методом – Immulate 1000, тропоніну (Siemens), біохімічним методом – загальну активність креатинфосфокінази (КФК ) (Cormay) та імуноферментним методом – ST2 «The Pressage ST2 Assay» (Сritical Diagnostics, № кат. BC-1065E). Детекцію білка Klotho, HSP70 білків, 8-OHdG, ST2 здійснювали на імуноферментному комплекcі Immuno Chem-2100 (США).Моделювання ГІМ в експериментальних тварин призводило до збільшення маркерів пошкодження серцевого м’яза тропоніну, активності загальної КФК. Важливо зазначити, що дослідні маркери мали тенденцію до збільшення в тварин усіх експериментальних груп порівняно з інтактною. На тлівведення вітаміну D і Necrostatin-1 значення ST2 не досягали рівня 35 нг/мл. Крім того, експериментальна терапія вітаміном D і Necrostatin-1 призводила до статистично значущого збільшення вмісту в гомогенаті серця білка Klotho (відповідно на 76,3 % і 78,8 % щодо контролю). Застосування індукторів синтезу білка Klotho сприяло обмеженню оксидативного пошкодження нуклеїнових кислот, що відображалось значним порівняно до контролю зниженням концентрації 8-OHdG як на тлі вітаміну D (на 38,5 %), так і Necrostatin-1 (на 55,8 %).Отримані експериментальні дані зумовлюють перспективність подальших досліджень у цьому напрямі. Особливий інтерес викликає дослідження здатності вітаміну D і Necrostatin-1 впливати на процеси клітинної загибелі та її тип (некроз/апоптоз).

Integrated ecotoxicological assessment of the complex interactions between chlorpyrifos and glyphosate on a non-target species Cnesterodon decemmaculatus (Jenyns, 1842)

Pesticide mixtures are frequent in freshwaters systems around the world, threatening the biota exposed to these conditions. The aim of this study was to determine the single and joint effect of two widely used pesticides in southern South America on a widely distributed fish species. In a 96-h assay, individuals of Cnesterodon decemmaculatus were exposed to 0.84 nL/L and 8.4 nL/L of Clorfox and 0.2 mg/L and 2 mg/L of Roundup Max, commercial formulations of chlorpyrifos and glyphosate, respectively. Also, there were four mixture treatments with all the possible combinations of both pesticides. A multi-level approach was carried out to assess their effects covering the following relevant biomarkers: behavior (immobile time, line crossings and average speed), somatic conditions (Fulton condition factor and hepatosomatic index), serum parameters (cortisol levels, lactate dehydrogenase (LDH), and creatine phosphokinase activity (CPK)), brain and muscle acetylcholinesterase and cytological characteristics (micronuclei frequency and nuclear abnormalities in erythrocytes). Our results showed that Clorfox exposures affect behavioral parameters, serum cortisol, and nuclear characteristics of erythrocytes. Roundup Max affects only the cortisol levels whereas mixture treatments have an effect on behavioral parameters, cortisol levels, LDH and CPK activities, and nuclear characteristics of erythrocytes. Potentiation was the main interaction at the lowest concentrations of both pesticides whereas antagonism occurred at the highest concentrations of both pesticides. These results are highly significant since they arise from an integrated ecotoxicological assessment at several levels of biological organization but even more important is that the potentiated effects of the mixtures we registered are environmentally relevant concentrations.

Predictive Factors for Muscle Injury After Posterior Lumbar Spinal Surgery

The aims of this study were to evaluate the risk factors for muscle injury in patients undergoing posterior lumbar spinal surgery and the clearance of postoperative biochemical changes following lumbar fusion and secondarily to evaluate the timing for monitoring postoperative biochemical serum levels and potential clinical correlation. The study prospectively enrolled 39 patients with degenerative disease of the lumbar spine. Biochemical markers (creatine phosphokinase [CPK], creatinine, and hemoglobin) were analyzed in 5 predefined stages. All relevant clinical data were collected. Rhabdomyolysis (RML) was defined as a postoperative 5-fold increase of the baseline CPK value. Patients from the lumbar fusion group had the highest postoperative CPK ratio. Overall, the rate of RML was 43.6%. CPK and creatinine activity reached their maximum on the first postoperative day in 69.2% and 87.5% of patients, respectively. Lumbar fusion (P= 0.005), surgical time >270 minutes (P= 0.028), and fall in hemoglobin levels >3 g/dL (P= 0.034) were identified as independent factors associated with higher risk of RML. The risk of RML increases with prolonged and invasive surgery with higher bleeding potential. Knowing the clearance of postoperative biochemical changes permits a standardized strategy with measurements in precise intervals, thereby avoiding unnecessary costs. The clinical significance is still undetermined.

High altitude mediated skeletal muscle atrophy: Protective role of curcumin

Chronic hypobaric hypoxia induced muscle atrophy results in decreased physical performance at high altitude. Curcumin has been shown to have muscle sparing effects under cachectic conditions. However, the protective effects of curcumin under chronic hypobaric hypoxia have not been studied till now. Therefore, the present study aims at evaluating the effects of curcumin administration on muscle atrophy under chronic hypobaric hypoxia. Male Sprague Dawley rats were divided into four groups: Control (C)-normoxia exposed, Control Treated (CT)-normoxia exposed and administered with curcumin at a dose of 100 mg/kg body weight for 14 days, Hypoxia (H)-exposed to hypobaric hypoxia for 14 days and Hypoxia Treated (HT)-exposed to hypobaric hypoxia and administered with curcumin for 14 days. Oxidative stress, muscle protein degradation, proteolytic pathways, myosin heavy chain (MHC), CPK activity and muscle histology were performed in gastrocnemius muscle samples of the exposed rats. In addition, fatigue time on treadmill running was also evaluated to observe the effects of curcumin administration on physical performance of the rats. As previously shown, hypobaric hypoxia increased muscle protein degradation via upregulated calpain and ubiquitin-proteolytic pathways. An enhanced oxidative stress has been linked to upregulation of these pathways under hypoxic conditions. Curcumin administration resulted in reduced oxidative stress as well as reduced activity of the proteolytic pathways in HT group as compared to H group thereby resulting in reduced muscle protein degradation under hypobaric hypoxia. Histology of rat muscle revealed an increased number of muscle fibres in HT as compared to H group. Thus, increased number of muscle fibres and decreased muscle proteolysis following curcumin administration, lead to enhanced muscle mass under hypobaric hypoxia resulting in improved physical performance of the rats.

Aortic thromboembolism with hind limb paralysis in a Japanese Black calf

A 7-day-old Japanese Black calf was presented to a local veterinarian with chief complaints of acute astasia, posterior paresis, and anorexia. Traumatic spinal injury was initially suspected. However, physical examination revealed hind leg coldness, absence of bilateral femoral artery pulse and deep pain sensation, diminished spinal reflexes, and dry gangrene of hind legs, suggesting thromboembolism of external iliac arteries. Increased activities of AST, LDH, and CPK were also suggestive of necrosis. On necropsy, a thrombus was found at the abdominal aortic bifurcation to bilateral external iliac arteries. Septic bacterial infection was suspected as the cause of aortic thromboembolism because Pseudomonas aeruginosa and Alcaligenes spp. were isolated from the thrombus. Aortic thromboembolism should be included in the differential diagnosis of posterior paresis.

Effects of selenium (Sel-Plex) supplement on blood biochemical parameters of juvenile Siberian sturgeon (Acipenser baerii)

Selenium (Se) and many of its compounds are among the most toxic micronutrients. Therefore, determining the optimal level of selenium in the diet of fish is one of the main concerns of researchers. The present study was designed to investigate the effects of Sel-Plex® (Selenium-enriched Saccharomyces cerevisiae yeast) on blood biochemical parameters in juvenile Siberian sturgeon, Acipenser baerii, as clinical biomarkers to evaluate the effects of selenium on the health of fish. In this study, fish were fed diets supplemented with 0 (control), 5, 10 and 15 g Sel-Plex per Kg feed for 8 weeks. Although a significant (P < 0.05) decrease was observed in alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT) activity in fish fed with 15 g Sel-Plex supplement, plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were significantly higher (P < 0.05) than the control fish. The results showed that administration of 10 and 15 g Sel-Plex supplement in fish significantly (P < 0.05) increased LDH and CPK activities. Administration of Sel-Plex significantly (P < 0.05) decreased glucose, albumin, cholesterol and triglyceride concentrations. The results revealed that Sel-Plex significantly increased (P < 0.05) globulin concentrations in the supplemented groups compared with the control group. Decreased total protein concentrations were only observed in fish fed with 5 g Sel-Plex. The results of this study show that diets containing high concentrations of Sel-Plex (10 g and 15 g) produced serious toxic effects, including changes in blood biochemical parameters.

Autophosphorylation-based Calcium (Ca2+) Sensitivity Priming and Ca2+/Calmodulin Inhibition of Arabidopsis thaliana Ca2+-dependent Protein Kinase 28 (CPK28)

Plant calcium (Ca2+)-dependent protein kinases (CPKs) represent the primary Ca2+-dependent protein kinase activities in plant systems. CPKs are composed of a dual specificity (Ser/Thr and Tyr) kinase domain tethered to a calmodulin-like domain (CLD) via an autoinhibitory junction (J). Although regulation of CPKs by Ca2+ has been extensively studied, the contribution of autophosphorylation in controlling CPK activity is less well understood. Furthermore, whether calmodulin (CaM) contributes to CPK regulation, as is the case for Ca2+/CaM-dependent protein kinases outside the plant lineage, remains an open question. We therefore screened a subset of plant CPKs for CaM binding and found that CPK28 is a high affinity Ca2+/CaM-binding protein. Using synthetic peptides and native gel electrophoresis, we coarsely mapped the CaM-binding domain to a site within the CPK28 J domain that overlaps with the known site of intramolecular interaction between the J domain and the CLD. Peptide kinase activity of fully dephosphorylated CPK28 was Ca2+-responsive and was inhibited by Ca2+/CaM. Using in situ autophosphorylated protein, we expand on the known set of CPK28 autophosphorylation sites, and we demonstrate that, unexpectedly, autophosphorylated CPK28 had enhanced kinase activity at physiological concentrations of Ca2+ compared with the dephosphorylated protein, suggesting that autophosphorylation functions to prime CPK28 for Ca2+ activation and might also allow CPK28 to remain active when Ca2+ levels are low. Furthermore, CPK28 autophosphorylation substantially reduced sensitivity of the kinase to Ca2+/CaM inhibition. Overall, our analyses uncover new complexities in the control of CPK28 and provide mechanistic support for Ca2+ signaling specificity through Ca2+ sensor priming.

КЛІНІКО-ФУНКЦІОНАЛЬНІ ТА БІОХІМІЧНІ АСПЕКТИ ЕФЕКТИВНОСТІ ЗАСТОСУВАННЯ КІНЕЗІОТЕЙПУВАННЯ У ВІДНОВНОМУ ЛІКУВАННІ ІДІОПАТИЧНОГО СКОЛІОЗУ

Метою дослідження було порівняти ефективності стандартного відновного лікування і додаткового залучення кінезіотейпування у дітей старшого шкільного віку, хворих на ідіопатичний сколіоз 1–2 ступенів. Проводили зіставлення показників динаміки клінічних, біомеханічних та рентгенологічних досліджень, ультразвукової денситометрії та біохімічного дослідження маркерів стану опорно-рухового апарату. Дослідження встановило, що застосування кінезіотейпування призводить до більш значних зсувів показників дослідження, чим при стандартному відновлювальному лікуванні, на що вказує суттєве переважання позитивної динаміки: рентгенографічних даних – зсувів кута викривлення хребта – (-1,18±0,86)о (при стандартному лікуванні – (-0,61±1,02)о, р&lt;0,0016); оцінки болю за ВАШ – (-1,66±1,25) бала (при стандартному лікуванні – (-0,29±0,58) бала, р&lt;0,0001); біомеханічних – згинання хребта – (-0,91±0,84) см (при стандартному лікуванні – (-0,32±0,50) см, р&lt;0,0001) і розгинання хребта – (-1,41±1,78) (при стандартному лікуванні – (-0,24±1,28)о, р&lt;0,0001). Також при застосуванні кінезіотейпування значно переважають зсуви активності у крові ЛФ, вмісту в крові креатиніну, активності у сечі КФК та вмісту в сечі оксипроліну і кальцію. Отже, застосування кінезіотейпування дозволяє підвищити ефективність відновного лікування ідіопатичного сколіозу 1–2 ступенів, що обґрунтовує подальші дослідження у цьому напрямку.

The effects of mebudipine on myocardial arrhythmia induced by ischemia-reperfusion injury in isolated rat heart

Reperfusion of the heart after an ischemic insult may lead to potentially lethal arrhythmias and cardimyocyte cell death via apoptosis and necrosis. In addition, previous studies showed that calcium channel blockers may have a protective role in the myocardium against arrhythmia and irreversible tissue injury. Therefore, this study was aimed to investigate the effects of mebudipine on myocardial arrhythmias and tissue injury induced by ischemia/reperfusion injury in isolated rat hearts. Male Wistar rats (250‑300g) were randomly divided to Sham group (without ischemia), control group (ischemia without drug), drug group (ischemia with mebudipine 0.1nM) and vehicle group (ischemia with ethanol 0.01%). The hearts of anaesthetized rats were removed and mounted on Langendorff apparatus and perfused by Krebs‑Henseleit solution under constant pressure of 75 mmHg at 37°C. Impulsive heart rate was monitored with bipolar golden electrodes. The electrocardiographs were recorded throughout the experiment and interpreted using the Lambeth convention. LDH and CPK activities in coronary effluent were analyzed spectrophotometrically. Hematoxilin &amp; Eosin staining was performed for evaluation of microscopic architecture of the myocardium and tissue injury. Pretreatment with mebudipine significantly decreased the number of ventricular premature beats (VPB) as compared with control group. The similar findings were seen in the number of ventricular tachycardia (VT) and fibrillation (VF) among groups. In addition, mebudipine significantly reduced the severity of arrhythmias in comparison with control hearts. Moreover, the drug group demonstrated marked improvement in edema and infiltration of inflammatory cells especially with regard to the degree of myonecrosis and cell lysis.Mebudipine diminished the number and the incidence of myocardial arrhythmias induced by reperfusion injury and the severity of tissue injury.

Creatine kinase isoform B distribution in the brain in schizophrenia

To compare patterns of brain isoform creatine phosphokinase (CPK B) distributions in post-mortem brain from patients with schizophrenia (Sch) and patients with somatic diseases (controls). Extracts of readily soluble and membrane-associated proteins were prepared from post-mortem samples of prefrontal cortex (Brodmann area 10), anterior (area 24) and posterior (area 23) cingulate cortex, hippocampus and cerebellum cortex from patients with Sch and control group (the samples were matched by age and postmortem interval). CPK enzymatic activity was measured by determination of inorganic phosphate, amounts of immunoreative CPK В were estimated by ECL-Western blotting using monoclonal antibodies. Asignificant decrease in CPK activity and amounts of immunoreative CPK В was observed in fractions of readily soluble proteins in all studied brain structures of patients with Sch compared to controls (p<0.01). Significant differences in CPK activity were found in membrane-associated protein fractions from the hippocampus (p<0.01), but not from the cingulate cortex (areas 23 and 24), of Sch patients compared with controls, whereas no difference between groups was found in levels of immunoreactive CPK B in membrane-associated protein fractions from the cingulate cortex (areas 23 and 24) and hippocampus. The decrease in the amount of CPK B in the frontal cortex of patients with Sch was confirmed by purification of CPK B active dimer from brain samples of patients with Sch and controls. Changes in the levels of CPK brain isoform in the brain of patients with Sch (the decrease in CPK activity and amounts in various brain structures at different extents) lead to the substantial alteration of CPK distribution pattern among the brain areas studied, result in the disturbance of the brain energy metabolism and contribute to Sch pathogenesis.

Astragaloside IV protects cardiomyocytes from anoxia/reoxygenation injury by upregulating the expression of Hes1 protein.

Astragaloside IV (ASI), a traditional Chinese medicine, is a main active ingredient of Astragalus membranaceus. Many clinical studies have found that ASI protects cardiomyocytes in cardiovascular diseases, but the underlying mechanisms remain obscure. The aim of this study was to investigate the molecular mechanisms responsible for the protective effects of ASI in cardiomyocytes from anoxia/reoxygenation (A/R) injury. According to the previous studies, we hypothesized that the cardioprotective effects of ASI against A/R injury might be associated with Notch1/Hes1 signaling pathway. In this study, neonatal rat primary cardiomyocytes were preconditioned with ASI prior to A/R injury. Our results showed that ASI effectively increased the cell viability, decreased the content of MDA, decreased the activities of CPK and LDH, increased the activities of GSH-Px and SOD, and reduced the reactive oxygen species (ROS) generation and the loss of mitochondrial membrane potential (Δψm). ASI inhibited the mitochondrial permeability transition pore (mPTP) opening and activation of caspase-3, and finally decreased the cell apoptosis in cardiomyocytes. Furthermore, ASI upregulated Hes1 protein expression. However, pretreatment with DAPT, a Notch1 inhibitor, effectively attenuated the cardioprotective effects of ASI against A/R injury, except MDA, SOD, GSH-Px, and the ROS generation. Taken together, we demonstrated that ASI could protect against A/R injury via the Notch1/Hes1 signaling pathway.

An evaluation of some oxidative and enzymatic biomarkers in different stages of naturally occurring copper poisoning in sheep

BACKGROUND: The early stage of Copper poisoning is difficult to be clinically diagnosed in sheep and has not been documented clearly yet. OBJECTIVES: To assess biomarkers in predicting early Copper poisoning in sheep, blood samples were collected from several groups of animals from a naturally Copper poisoning occurrence in an industrial region. METHODS: Animals were divided into four experimental groups; Group A: far from polluted region (Control group), Group B: inside polluted region, apparently healthy and without any clinically jaundice manifestation, Group C: slightly with jaundice signs and hemolytic crisis phase, and Group D: with clear jaundice signs. After collecting blood samples from each group, the serum was analyzed for evaluation of liver enzymes and oxidative stress parameters in different stages of Copper poisoning. In each blood sample, CPK, GGT, AST, ALT, total thiol (T-SH) group, and total proteins were determined. The Copper concentration in the serum, liver, and kidney of the dead animals in group D were also determined. RESULTS: There were significant differences in the blood parameters in group C illustrated by elevated level of serum AST, CPK, and GGT activities and total thiol (as biomarker of oxidative stress) when compared to control groups. In group D, these enzymes, in addition to T-SH, and the total protein were significantly (p≤ 0.05) different from those of the control and the other groups. Measurement of Copper in serum, liver, and kidney of group D (at the end stage of hemolytic phase) confirmed Copper poisoning in these groups. CONCLUSIONS: Based on the findings of the present study, the measurement of the liver enzyme activities and total thiol just closed to critical hemolytic phase could be reliable biomarkers for predicting Copper poisoning in sheep.

Physiological and Histological Studies on the Heart of Male Albino Rats Exposed to Electromagnetic Field and the Protective Role of Silymarin and/Orvitamin E.

Aim of the work:This study aimed to determine the ameliorative effect of silymarin (SIL) and\or vitamin E (Vit.E) against changes induced by mobile phone radiation in the heart of male albino rats. Matrerial and methods:Total of 48 adult male albino rats were assigned for this study. The 1st group served as control (n=6), the 2nd group exposed to mobile phone generator radiation (900MHz) for 2hr/day 3days/week for two months, 3rd group (+ve control) supplemented with SIL, 4thgroup (+ve control) supplemented with Vit. E, 5th group (+ve control) supplemented with SIL and Vit.E, 6th group: exposed group supplemented with SIL, 7th group: exposed group supplemented with Vit.E and 8th group exposed group supplemented with SIL and Vit.E. Physiological ,histopathological and histochemical changes were studied.Results: Exposure to mobile phone causes increases in activities of CPK, CK-MB and LDH enzymes in serum and heart tissue and oxidative stress markers (MDA and H2O2),while antioxidants (CAT and GSH) were decreased in the heart tissue. Sodium (Na) and calcium (Ca) levels were decreased While, K level showed non-significant change in serum. Numerous histopathological changes were detected in the heart tissue of rats of the irradiated group with altered collagen fibres, polysaccharides in the cardiac muscle fibres of the exposed group. These changes manifested good amelioration in the exposed groups that supplemented with SIL and/or Vit.E. Conclusion:Treatment of rats with SIL and/or Vit.E ameliorated the dangerous effect of mobile phone radiation occurred in the cardiac muscle fibres.

Abstract 15175: Critical Role of Sodium-Glucose Cotransporter 1 in the Cardioprotection Through the Maintenance of Energy Production During Ischemia-Reperfusion Injury

Glucose becomes an important preferential substrate for cardiac metabolism and ATP generation under specific pathological conditions, such as ischemia. Glucose utilization is initiated by glucose uptake, which is mainly controlled by glucose transporters. Although the role of passive glucose transporters (GLUTs) has been investigated intensively, little is known about the functional significance of the other active transporter, sodium-glucose cotransporter 1 (SGLT1), in the heart. We herein hypothesized that cardiac SGLT1 plays a critical role for cardioprotection during ischemia-reperfusion injury (IRI), possibly through the enhanced glucose utilization for the energy production. To test this, we studied the effects of 10-4 M of phlorizin, SGLT1 inhibitor, on the baseline function and its response to global ischemia (20 minutes)-reperfusion (40 minutes) injury in mouse Langendorff perfusion model, where SGLT1, but not SGLT2, is highly expressed, as confirmed by immunoblotting assay. Although phlorizin did not affect baseline function, its administration during IRI significantly impaired recovery in left ventricular contraction (% recovery of baseline; 67.3±4.5 vs. 89.7±6.8%, n=5 each, P&lt;0.05) and rate pressure product (18600±1290 vs. 25100±1010 mmHg•bpm, n=5 each, P&lt;0.01), associated with increased infarct size demonstrated by TTC staining as well as CPK activity released into the perfusate (20.8±9.5 vs. 2.0±0.6 U•g, n≥3, P&lt;0.05). Of note, the onset of ischemic contracture, which is thought to be initiated by ATP depletion in cardiomyocytes, was earlier with phlorizin perfusion (288±21 vs. 364±27 sec, n≥8, P&lt;0.05). Consistent with this, the significant reduction of tissue ATP content was observed in phlorizin perfused heart (4.81±0.61 vs. 6.87±0.13 μmol/g tissue, n≥4, P&lt;0.05). In conclusion, these data demonstrate that SGLT1 represents an important cardioprotective mechanism against IRI possibly through the maintenance of ATP generation. Our findings shed new light on the essential role of SGLT1 for the optimization of cardiac energy metabolism by the enhanced glucose availability during ischemia, thus leading to substantial myocardial protection after severe ischemic insults.

Search of substances with the cardioprotective activity in the series of derivatives of halogenides 1-alkyl- and carboxyalkyl-4-ylideneamino-1,2,4-triazole

In spite of achievements in the sphere of highly effective drugs creation for treating cardiovascular diseases the problem is still urgent since mortality is high because of these diseases, and it takes the 2-nd – 3-d place among the population of industrially developed countries. In the Research and Production Association “Pharmatron” a new approach concerning creation of new effective anti-anginal drugs has been developed; it is a chemical modification of position 1 and 4 of 1,2,4-triazole molecule by introduction of the structural fragments of the most active medicines and structures imitating sites of adrenergic receptors. Bromides of 1-(alkyl) carboxyalkyl-4-ylideneamino-1,2,4-triazole have been synthesized at the Pharmaceutical Chemistry department of Zaporizhzhia State Medical University under the supervision of professor I.A.Mazur. The cardioprotective activity of 12 compounds – derivatives of 1-carboxyalkyl-4-ylideneamino-1,2,4-triazole bromides and 1-alkyl-4-ylidenamino-1,2,4-triazole bromides has been investigated. The experiments were conducted in 510 white outbred male rats weighing 120-130 g from the mouse bank of the Institute of Pharmacology and Toxicology of the AMS of Ukraine. Determination of acute toxicity of the compounds investigated was conducted according to Kerber method. It has been found that LD50 of these compounds when introduced intraabdominally to rats is within 29.5-295 mg/kg. It allows to refer the compounds under research to the III and IV classes of toxicity (moderate or low toxic). The model of acute myocardial infarction in rats was used for studying the cardioprotective activity of the compounds, the infarction was modeled by staged introduction of isadrin and pituitrin. It has been found that intraabdominal introduction of 8 compounds of 12 ones in the dose of 1/100 LD50 to rats with myocardial infarction leads to decrease of hyperenzymemia of cardiospecific isoenzymes of creatine phosphokinase (MB CPK) and lactate dehydrogenase (LDH-1), which play the role of biochemical markers of the myocardium damage, and decrease of electrophysiological marker ΣΔST during electrocardiography. The structural fragments of the molecule, which play a determinative role in manifestation of the cardioprotective effect of such series as β-phenylethyl, carboxy propyl, octyl in position 1 and the presence of amino- and n-methoxybensilidenamino-groups in position 4, have been revealed. It has been determined that compound MT significantly exceeds the therapeutical efficacy of metoprolol by decrease of MB CPK and LDH-1 activity in the blood serum of rats with myocardial infarction, as well as decrease of ECG ST amplitude.

Effect of different doses of uranyl acetate on some blood parameters and hepatic enzymesin females rats.

This study was carried out to investigate effects of oral administration of uranyl acetate on haematological parameters and liver enzyme. For this purpose, twenty females mature Albino rats were divided into four equal groups, control group G1 administrated distilled water, the other three groups administrated orally 50,75,100 mg/kg/b.w. /day of uranyl acetate. The route of administration was oral intubations for 10 weeks, for 14 days before mating with untreated males, as well as during pregnancy 3 weeks and lactation 5 weeks. At the end of the treatment blood samples were collected from mothers of rats. The results were obtained a significant P&lt; 0.05 increase in the total WBC counts in groups G2,G3,G4 compared with control .A significant P&lt;0.05 decrease in the lymphocytes and monocytes percentage in groups (G3,G4), while no significant decrease P&gt;0.05 in G2 in the lymphocytes and monocytes percentage compared with control. And the results show significant P&lt; 0.05 decrease in HB value in groups G3,G4, while no significant decrease P&gt;0.05 in G2 in HB value compared with control. A significant P&lt; 0.05 decrease was observed in total RBCs counts in groups G2,G3,G4 compared with control. No significant increase P&gt;0.05 in GOT and GPT in groups G2 and G3 and significant P&lt;0.05 increase in GOT and GPT activity in the serum in group G4 compared with control. No significant increase P&gt;0.05 in CPK activity in groups G2,G3,G4 compared with control.