Increased Creatine Kinase Activity Research Articles

Effects of Ashwagandha (Withania Somnifera) Supplementation on Body Composition and Blood Health Indices in Professional Wrestlers

Abstract Introduction. The purpose of this randomized, double-blind, placebo-controlled study was to evaluate the effect of ashwagandha supplementation on the body composition, blood health and recovery indices, as well as the safety of ashwagandha supplementation in wrestling. Material and Methods. To investigate this, body composition (using dual energy X-ray absorptiometry; DEXA) and blood parameters as indicators of athletes’ health status (i.e., hematology, liver, kidney and muscle tissue function, lipid profile) were measured before and after 8-week supplementation with ashwagandha extract (600 mg/day) in professional national team wrestlers following an intensive training plan (as part of an annual training cycle). Results. At the end of the study, there was a significant increase in fat-free mass and muscle mass, with a significant decrease in fat mass (both in absolute and relative terms) in both ashwagandha and placebo groups (main effect of time, p < 0.05). However, ashwagandha supplementation did not affect any of the body composition parameters (main effect of group or interaction of time and group, p > 0.05). Moreover, intense wrestling training induced an increase in blood creatine kinase activity as an indicator of muscle tissue damage (main effect of time, p < 0.05). Ashwagandha supplementation did not significantly affect any hematological or biochemical blood parameters (p > 0.05 for main effect of group and interaction of time and group). However, the increase in creatine kinase activity during the study seemed to be more pronounced in the placebo group (post-hoc; p < 0.05) than in the ashwagandha group (post-hoc; p > 0.05). Conclusions. In conclusion, ashwagandha supplementation at 600 mg daily for 8 weeks is well tolerated and safe for professional wrestlers. This strategy does not appear to offer additional body composition benefits beyond the effect of intense wrestling training. On the other hand, our results indicate some potential for ashwagandha in improving muscle tissue recovery, which needs to be confirmed on a larger population of athletes.

Medical causes of poor performance and their associations with fitness in Standardbred racehorses.

Poor performance is a multifactorial syndrome of racehorses, commonly associated with subclinical disorders, which can be diagnosed by exercise testing. Describe the prevalence of medical causes of poor performance in Standardbreds unassociated with lameness, and evaluate their relationships with fitness variables measured by exercise treadmill test. Hospital population of 259 nonlame Standardbred trotters referred for poor performance. The horses' medical records were retrospectively reviewed. Horses underwent a diagnostic protocol including resting examination, plasma lactate concentration, treadmill test with continuous ECG and assessment of fitness variables, creatine kinase activity, treadmill endoscopy, postexercise tracheobronchoscopy, bronchoalveolar lavage (BAL), and gastroscopy. The prevalence of different disorders was evaluated, including cardiac arrhythmias, exertional myopathies, dynamic upper airway obstructions (DUAOs), exercise-induced pulmonary hemorrhage (EIPH), moderate equine asthma (MEA), and gastric ulcers (EGUS). The associations of these disorders with fitness variables were investigated individually and using multivariable models. Moderate equine asthma and EGUS were the most common disorders, followed by EIPH, DUAOs, cardiac arrhythmias, and exertional myopathies. Hemosiderin score was positively correlated with BAL neutrophils, eosinophils, and mast cells; increased creatine kinase activity was associated with BAL neutrophilia, DUAOs, premature complexes, and squamous gastric disease. Treadmill velocity at a plasma lactate concentration of 4 mmol/L and at heart rate of 200 beats per minute was negatively affected by BAL neutrophilia, multiple DUAOs, exertional myopathies, and squamous gastric disease. The multifactorial nature of poor performance was confirmed, with MEA, DUAOs, myopathies and EGUS representing the main diseases involved in fitness impairment.

Changes in Skeletal Muscle Troponin T and Vitamin D Binding Protein (DBP) Concentrations in the Blood of Male Amateur Athletes Participating in a Marathon and 100 km Adventure Race

This study assessed changes in creatine kinase (CK) activity and skeletal muscle troponin T (sTnT) concentrations in the blood, to estimate the degree of muscle degradation after exercise. In addition, the concentration of vitamin D binding protein (DBP) in the blood was assessed. DBP concentrations were measured in blood as a marker for plasma load by monomeric actin. The study included marathon (MR) participants and 100 km adventure race (AR) participants, who were examined before and after the race. There was a significant (16-fold) increase in CK activity among AR participants, and a significant increase in sTnT concentration-127% in the MR group and 113% in the AR group, while there was a statistically significant decrease in DBP concentration by 14% in the AR group. In addition, it was observed that the initial concentration of DBP in both groups was in a normal range, but was lower than the average population, and the DBP concentration in the AR group was lower than in the MR group. It was concluded that exhausting physical effort such as a marathon or adventure races causes muscle damage with a far stronger influence on sarcoplasm than on filaments. The short-term and slight reduction in the concentration of DBP in blood after such efforts may be due to the appearance of monomeric actin in plasma.

Clinical manifestations, laboratory findings, treatment and outcome of acute organophosphate or carbamate intoxication in 39 cats.

Organophosphates and carbamates are important sources of intoxication for humans and animals. However, large-scale studies of these intoxications in cats are unavailable. The medical records of 39 cats presented to a veterinary teaching hospital with acute organophosphate or carbamate intoxication were reviewed retrospectively. Mortality in intoxicated cats was 15%. Low respiratory rate and low rectal temperature at presentation were associated with death. Other common clinical signs included weakness, ataxia, apathy, recumbency, anorexia and bradycardia, but these were unassociated with the outcome. The common biochemical abnormalities included decreased serum butyryl-choline esterase activity, acidaemia, hypercarbaemia and total hypocalcaemia, and increased creatine kinase activity and total plasma protein concentration. There were no significant differences in haematological, biochemical and blood gas analytes between survivors and non-survivors. Common medications and treatments included 2-pyridine aldoxime methyl-chloride-pralidoxime (2-PAM) (74%), metoclopramide (64%), antibiotics (64%), diphenhydramine (59%) and atropine sulphate (54%). There were no significant drug and treatment differences between survivors and non-survivors. The secondary complications of the intoxication included pneumonia (10%), acute kidney injury (10%) and pancreatitis (8%). Acute cholinergic crisis due to organophosphate or carbamate intoxication has a fair prognosis in cats. Low respiratory rate and low rectal temperature at presentation were associated with death. The most commonly used specific medications in this study included 2-PAM, diphenhydramine and atropine sulphate.

Pre-race and race management impacts serum muscle enzyme activity in Australian endurance horses.

BackgroundMarked increases in serum muscle enzyme activity can occur in endurance horses but the diagnostic certainty in predicting cases of myopathy is unclear. Improved understanding of horse management effects on serum muscle enzyme activity as markers of muscle health would assist interpretation of serum muscle enzyme activity and guide management to reduce myopathy risk.ObjectivesTo investigate associations between serum muscle enzyme activity and management factors in endurance horses.Study designCross‐sectional study.MethodsOne hundred endurance horses competing in four endurance events (offering distances of 20‐120 km) in south‐eastern Australia were observed. Data were collected from official horse logbooks, pre‐and post‐race serum samples, an owner questionnaire of pre‐race and race management of horses and the Australian Endurance Riders Association results database. Multivariable linear regression modelling tested associations between management factors and serum muscle enzyme activity.ResultsFirst leg speed, distance raced, number of rest days pre‐race, and pre‐race activity of aspartate aminotransferase (AST) and creatine kinase (CK) explained 47.3% of the variance in post‐race CK. As first leg speed increased by 1 km/h, CK activity increased by 25.8% (95% CI 11%‐35%). Race distances >80 km increased post‐race CK activity by 124% (95% CI 116%‐145%). Each additional pre‐race rest day increased post‐race CK activity by 30.5% (95% CI 11%‐42%). Modelling a 10% increase in pre‐race CK and pre‐race AST activity was associated with post‐race CK activity increasing by 7.3% (95% CI 3%‐14.4%) and 8.5% (95% CI 0.3%‐14.2%) respectively. Horses experiencing training distances >40 km and a greater number of rest days prior to race day developed increased pre‐race AST and CK activity respectively.Main limitationsOwner questionnaires may be subject to bias. Limited data were available to model ride terrain, horse fitness, ration detail and myopathy. Muscle biopsies were not used to confirm myopathy.ConclusionsNearly half of the variation in post‐race CK activity observed can be attributed to management factors unrelated to myopathy, suggesting increased CK activity may not be pathognomonic for myopathy. We advise caution in relying solely on serum muscle enzyme activity for diagnosis of myopathy until the strength of association between CK and myopathy is further ascertained in future studies.

The Effect of Grape Seed Extract Supplementation on Delayed Onset Muscle Soreness (DOMS) in Young Healthy Female Students

Background: Delayed onset muscle soreness (DOMS) is a common experience after unusual activities particularly eccentric exercises. The aim of the present study was to investigate the effect of grape seed extract (GSE) on some factors of DOMS on non-athletic female students. Method: Twenty young healthy female students were randomly allocated into two experimental (GSE=10n) and placebo (PLC=10n) groups. T produce DOMS, a forty-five repetition of eccentric elbow curl in three sets was performed. Venous blood samples were obtained before and after supplementation, immediately, 24h, and 48h after exercise for analysis of creatine kinase (CK), lactate dehydrogenase (LDH). The pain was assessed by visual analogue scale (VAS). Results: There was a significant increase in CK activity after exercise compared with before exercise in both groups (P<0.05)‚ but no significant differences between two groups at any time (P>0.05). Furthermore, The VAS scores were significantly lower in GSE compared with PLC group 24h and 48h after exercise (P<0.05). Conclusion: Although GSE supplementation has not been able to affect the CK and LDH activity, it seems that its anti-inflammatory or analgesic effects have been reduced the pain caused by DOMS.

Vasoplegia induces high energy demand and depletes cardiac energetics: a new insight into physiological concepts of cardiac work and a model for acute heart failure?

Abstract Funding Acknowledgements Type of funding sources: Other. Main funding source(s): British Heart Foundation Background / Introduction Hearts subjected to constant high workload will, over time, fail.  We hypothesized vasoplegia would create a high cardiac energy demand and deplete cardiac energy reserves, a mechanism causing failure. With 31Phosphorus Magnetic Resonance Spectroscopy, we can measure energy: ATP usage (via creatine kinase flux) and reserves of phosphocreatine (PCr/ATP ratio). Methods: Ten healthy volunteers were recruited mean age 37, mean BMI 22.4, 5 male, 5 female.  A nitrate vasodilator (Glyceryl trinitrate, GTN) was used to induce vasoplegia, with MRI and MRS measurements taken before and during. Results There was reduced preload (falls in end diastolic volume and right atrial area) and afterload (mean blood pressure fall 14mmHg, p <0.0001).  Subjects mounted reflex tachycardia (mean heart rate rise 9bpm, p < 0.0001) and inotropy (mean baseline ejection fraction increase 5%, p = 0.0002) hence cardiac output and rate pressure product were maintained (baseline CO 6.24 ± 1.45 L/min vs GTN 6.49 ± 1.43 L/min, p = 0.37; baseline RPP = 6929 ± 976 mmHg.bpm vs GTN 7214 ± 1051 mmHg.bpm, p = 0.06). There was a 58% increase in Creatine Kinase activity (kf) (baseline 0.158 s-1 vs GTN 0.249 s-1, p = 0.006) and CK flux (baseline 1.79 ± 0.78 umol/g/s vs GTN 2.59 ± 1.07 umol/g/s, p = 0.03), demonstrating increased ATP transfer.  PCr/ATP fell (from 2.17 ± 0.2 to 1.99 ± 0.22, p = 0.027).  Baseline PCr/ATP positively correlated with the change in stroke volume (R2 = 0.66, p = 0.005) during administration of GTN. In addition, whilst at rest, CK kf correlated with rate pressure product (R2 = 0.56, p = 0.03), there was no correlation seen during GTN.   Conclusions  We have shown vasoplegia increases energy demand upon the heart, despite unchanged cardiac output and rate pressure product.  Further, we have shown a fall in Phosphocreatine/ATP ratio, suggesting this ATP transfer outstripped oxidative phosphorylation and the phosphocreatine pool was depleted in order to maintain ATP delivery.  We speculate that progressive depletion of energetics in the context of high energy demand drives hearts under constant stress into failure. In addition, we demonstrate that the magnitude of the resting phosphocreatine pool relates to ability to augment stroke volume during stress and suggest that this may be protective against a decline into failure.  Further work is needed, but PCr/ATP may be an important biomarker for treatments that target myocardial energetics to protect against failure. Abstract Figure 1

The phosphoenolpyruvate carboxykinase (PEPCK) inhibitor, 3-mercaptopicolinic acid (3-MPA), induces myogenic differentiation in C2C12 cells

Phosphoenolpyruvate carboxykinase (PEPCK) is a gluconeogenic enzyme with a cytosolic (Pck1/PEPCK-C) and mitochondrial (Pck2/PEPCK-M) isoform. Here we investigate the effect of 3-mercaptopicolinic acid (3-MPA), a PEPCK inhibitor, on C2C12 muscle cells. We report that Pck2 mRNA is 50–5000-fold higher than Pck1 during C2C12 myogenesis, indicating Pck2 is the predominant PEPCK isoform. C2C12 cell proliferation was inhibited in a dose-dependent manner following 48 h 3-MPA treatment (0.01–1 mM). C2C12 myogenic differentiation was significantly induced following 3-MPA treatment (0.25, 0.5, 1 mM) from day 0 of differentiation, demonstrated by increased creatine kinase activity, fusion index and myotube diameter; likewise, the myosin heavy chain (MyHC)-IIB isoform (encoded by Myh4) is an indicator of hypertrophy, and both porcine MYH4-promoter activity and endogenous Myh4 mRNA were also significantly induced. High doses (0.5 and/or 1 mM) of 3-MPA reduced mRNA expression of Pck2 and genes associated with serine biosynthesis (Phosphoglycerate dehydrogenase, Phgdh; phosphoserine aminotransferase-1, Psat1) following treatment from days 0 and 4. To conclude, as Pck2/PEPCK-M is the predominant isoform in C2C12 cells, we postulate that 3-MPA promoted myogenic differentiation through the inhibition of PEPCK-M. However, we were unable to confirm that 3-MPA inhibited PEPCK-M enzyme activity as 3-MPA interfered with the PEPCK enzyme assay, particularly at 0.5 and 1 mM.

ENZYME DIAGNOSTICS IN SPORTS PRACTICE (LITERATURE REVIEW AND PERSONAL EXPERIENCE)

Aim. The paper aims to analyze and summarize the data about enzyme diagnostics in athletes of different sports. Materials and methods. Creatine kinase (CK) activity was measured in 20 highly skilled male biathletes aged from 18 to 23 years. Fasted blood samples were taken in the morning before exercise and the next day after a 90-minute ski run by a uniform method. Biochemical analysis was performed by using the Architect c4000 and i1000SR (Abbott, USA) automated analyzers with reagents and control materials from the equipment manufacturer. Results. An increase in CK activity in biathletes is observed at the end of the preparatory period, and normalization occurs by the middle of the competitive period. When studying CK activity in 10 male handball players aged from 18 to 21 years the day after training, it was found that the increase in enzyme activity depended on the playing position. Goalkeepers exceeded the norm many times compared to field players. The highest serum CK activity after exercise was observed after prolonged exercise such as extra-long marathon and uphill running, which involved eccentric muscle contractions. In triathletes after competitions (full distance), CK activity increased to 1200 U/L, after 48 hours of recovery it dropped to 400 U/L. Conclusion. Thus, enzyme diagnostics today is a relevant and objective method for assessing the state of health and the muscular system, including organs such as the heart, liver, and brain. Therefore, control over the level of basic enzymes should be carried out regularly in the preparatory period both as part of an in-depth medical examination and if an athlete is suspected of being overtrained or injured.

Effects of photobiomodulation therapy on the local experimental envenoming by Bothrops leucurus snake

Bothrops leucurus is the major causative agent of snakebites in Brazil's Northeast. The systemic effects of its venom are effectively neutralized by antivenom therapy, preventing bitten patients' death. However, antivenom fails in neutralizing local effects that include intense pain, edema, bleeding, and myonecrosis. Such effects can lead to irreversible sequels, representing a clinically relevant issue for which there is no current effective treatment. Herein, the effects of photobiomodulation therapy (PBMT) were tested in the local actions induced by B. leucurus venom (BLV) in mice (n = 123 animals in 20 experimental groups). A continuous emission AlGaAs semiconductor diode laser was used in two wavelengths (660 or 780 nm). Mechanical nociceptive thresholds were assessed with the electronic von Frey apparatus. Local edema was determined by measuring the increase in paw thickness. Hemorrhage was quantified by digital measurement of the bleeding area. Myotoxicity was evaluated by serum creatine kinase (CK) activity and histopathological analysis. PBMT promoted anti-hypernociception in BLV-injected mice; irradiation with the 660 nm laser resulted in faster effect onset than the 780 nm laser. Both laser protocols reduced paw edema formation, whether irradiation was performed immediately or half an hour after venom injection. BLV-induced hemorrhage was not altered by PBMT. Laser irradiation delayed, but did not prevent myotoxicity caused by BLV, as shown by a late increase in CK activity and histopathological alterations. PBMT was effective in the control of some of the major local effects of BLV refractory to antivenom. It is a potential complementary therapy that could be used in bothropic envenoming, minimizing the morbidity of these snakebite accidents.

Protective effects of diet containing rutin against trichlorfon-induced muscle bioenergetics disruption and impairment on fatty acid profile of silver catfish Rhamdia quelen

Trichlorfon is an organophosphate insecticide that is widely used on fish farms to control parasitic infections. It has been detected in freshwater ecosystems as well as in fishery products. There is a growing body of evidence to suggest that certain feed additives may reduce or prevent pesticide-induced toxicity in fish. The aim of the present study was to determine whether acute exposure to trichlorfon would alter bioenergetic homeostasis and alter fatty acid profiles in muscles of silver catfish (Rhamdia quelen). We also sought to determine whether rutin prevents or reduces these effects. Cytosolic and mitochondrial creatine kinase (CK) and activities of complexes II-III and IV in muscle were significantly inhibited by exposure to 11 mg/L trichlorfon for 48 h compared to effects in the unexposed group. Total content of polyunsaturated fatty acids (omega-3 and omega-6) were significantly lower in muscle of silver catfish exposed to 11 mg/L trichlorfon for 48 h than in the unexposed group. Addition of 3 mg rutin/kg feed increased CK activity and prevented inhibition of complex IV activity, as well as preventing all alterations of muscle fatty acid profiles elicited by exposure to trichlorfon. No significant differences were observed between groups with respect to muscle adenylate kinase or pyruvate kinase activities, as well as total content of saturated and monounsaturated fatty acids. Our findings suggest that exposure (48 h) to 11 mg trichlorfon/L water inhibits cytosolic and mitochondrial CK activity in muscle. Trichlorfon also affects activities of complexes II-III and IV in respiratory chain, with important consequences for adenosine triphosphate production. The pesticide alters fatty acid profiles in the fish and endangers human consumers of the product. The most important finding of the present study is that inclusion of rutin improves bioenergetic homeostasis and muscle fatty acid profiles, suggesting that it reduces trichlorfon-induced muscle damage.

Snake venomics, experimental toxic activities and clinical characteristics of human envenomation by Bothrocophias myersi (Serpentes: Viperidae) from Colombia

Venoms of the viperid genus Bothrocophias, restricted to Colombia and Ecuador, are poorly known. Only a proteomic analysis of B. campbelli venom has been described. In this work we present a proteomic study of B. myersi venom, its biological activities, and describe the clinical characteristics of a patient bitten by this species. B. myersi venom mainly consists of phospholipases A2 (54.0%) and metalloproteinases (21.5%), among proteins of twelve different families. This venom exhibited proteolytic, phospholipase A2, myotoxic, edema-forming, and lethal activities. Enzymatic activities did not show statistically significant differences in comparison to Bothrops asper venom, but B. myersi venom displayed weaker hemorrhagic and coagulant activities. Polyvalent Viperidae antivenoms produced in Costa Rica and Colombia cross-recognized B. myersi venom by ELISA, however only the latter neutralized its lethal activity in mice when tested at a ratio of 3 mg venom/mL antivenom, suggesting it should be useful to treat envenomings inflicted by this species. A patient bitten by B. myersi developed edema and myotoxicity, evidenced by an increased creatine kinase activity in plasma. A good correlation was found between experimental biological activities of Bothrocophias myersi venom and the clinical features of an envenoming provoked by this species. SignificanceThe proteomic characterization, toxicity, immunorecognition and neutralization of Bothrocophias myersi venom have been determined for the first time. The distribution of this pit viper is restricted to Colombia and Ecuador, and its venom contains a high proportion of phospholipases A2 and metalloproteinases. The polyvalent antivenom produced in Colombia neutralized the lethal activity of this venom in vivo, and therefore should be effective in the treatment of envenomings by this snake.

Creatine kinase/α-crystallin interaction functions in cataract development

Creatine kinase (CK) is an energy storage enzyme that plays an important role in energy metabolism. CK/phosphocreatine functions as an energy buffer and links ATP production sites with ATP utilization sites. Several key mutations in the αA-crystallin (cryaa) and αB-crystallin (cryab) genes have been linked with autosomal-dominant, hereditary human cataracts. The cryaa-R49C mutation was identified in a four-generation Caucasian family. We previously identified an increase in the quantity of CK complexed with α-crystallin in the lenses of knock-in mice expressing the cryaa-R49C mutation using proteomic analyses. Increased levels of CK in postnatal cataractous lenses may indicate increased ATP requirements during early cataract development. To gain a further understanding of the relationship between CK and α-crystallin, we investigated whether α-crystallin interacts with and forms complexes with CK, in vitro. Isothermal titration calorimetry (ITC) showed that each CK dimer bound to 28 α-crystallin subunits, with a Kd of 3.3 × 10−7 M, and that the interaction between α-crystallin and CK was endothermic, thermodynamically favorable, and entropy-driven. High-salt concentrations did not affect the interaction between CK and α-crystallin, suggesting that the interaction between CK and α-crystallin is primarily hydrophobic. Gel permeation chromatography (GPC) detected water-soluble α-crystallin and CK complexes, as determined by increased light scattering after complex formation. In addition, CK and α-crystallin formed partially-water-insoluble, high-molecular-mass complexes. Enzyme-linked immunosorbent assay (ELISA)-based enzymatic activity analyses of lens homogenates showed a 17-fold increase in CK activity in the postnatal lenses of cryaa-R49C knock-in mice. These studies indicate that the interaction between α-crystallin and CK is functionally important and that increased CK levels may be necessary to meet the increased ATP demands of ATP-dependent functions in cataractous lenses.

FORECAST OF THE OUTCOME OF SURGICAL TREATMENT OF PATIENTS WITH AORTIC ANEURISM TO CHANGE THE CONCENTRATION OF VESSEL ENDOTHELIUM GROWTH FACTOR (VEGF) AND CONTENT OF IMMUNOLOGICAL MARKERS

Summary. Despite all the successes of surgical treatment of patients with aortic aneurysm, there are a number of questions, the search for answers to which remains relevant now. It is important to have prognostic markers for assessing postoperative complications.
 The aim. To determine the diagnostic and prognostic significance of indicators characterizing immunoresistance, the degree of adaptive reactions and the regenerative ability of immunophysiological factors affecting the outcome of the disease.
 Materials and methods. The study examined 23 patients with vascular pathology (13 patients with aortic aneurysm (chest and abdominal) and 10 patients with coronary heart disease (CHD)).
 Results and discussion. The determination of significant biomarkers in patients with aortic aneurysms revealed violations of the barrier function of neutrophil phagocytosis. An increase in the oxidative activity of SP, % in the spontaneous NBT-test to (20,2 ± 2,2)% at a reference value of (10 ± 1,1)% and a 1,2-fold increase in the number of cells that absorbed the dye after stimulation with zymosan (ST). In all examined patients with aortic aneurysm, CD2+ and CD3+ stimulation was observed. The decrease in CD4+ and CD8+ in 1,7 and 1,6 times, respectively, in all terms of the survey. A significant increase in cytotoxic factors opsonized by the C3 complement factor by 1,5 times was revealed. In the preoperative period revealed a multiple increase in the concentration of IL-6 (in 60 times) and IL-10 (in 50 times) in all patients with aortic aneurysm. In 3 patients with negative dynamics of the course of the disease (exitus), an extremely low level of VEGF concentration was detected. In all patients with stratified aortic aneurysms, preoperatively reduced AChE activity was observed, an increase in creatinekinase activity to 410 U/l at a reference value of (85 ± 54) U/l, and a decrease in ceruloplasmin content to 214.4 mg/l compared to the reference group (315.0 ± 45.2) mg/l.
 Conclusion. As significant biomarkers characterizing the severity of the condition of patients with aortic aneurysm, the level of vascular endothelial growth factor (VEGF), cytokines, as well as changes in the activity of the enzyme acetylcholinesterase (AChE) can be used.

Semen Creatine and Creatine Kinase Activity as an Indicator of Sperm Quality.

The role of creatine (Cr) and creatine kinase (CK) in sperm function remains unclear. The study aimed to assess Cr and CK in seminal plasma and test their association with sperm characteristics. The study included 62 males with couple's infertility and 26 males who have already fathered children. Semen Cr and CK were assessed by GC-MS and spectrophotometry, respectively. Seminogram parameters were analyzed using conventional methods. Cytomorphologic analysis of sperm showed normozoospermia in 53 men (NS) and an asthenozoospermia (AS) in 35 men. Semen Cr was high with no significant difference between the two groups (791 ± 342 and 744 ± 422 µmol/L, respectively). However, semen CK activity was higher in AS group (1,360 ± 1,050 vs. 830 ± 580 U/L, p = 0.013). Semen Cr was positively related to progressive motility (r = 0.284; p = 0.010). Semen CK was negatively correlated with sperm concentration (r = -0.29; p = 0.01), progressive motility (r = -0.26; p = 0.03), and the percentage of abnormal spermatozoa (r = -0.28; p = 0.02). Semen contains high amounts of Cr and increased CK activity. Low semen Cr is associated with reduced sperm motility while high CK activity is associated with poor sperm quality. The findings suggest that Cr is of importance for sperm metabolism and that Cr supplementation could be useful in males with poor quality sperm.

Investigation into the Possible Risk Effects Associated with Drug and Alcohol-induced Gastrointestinal Ulcer in Wistar Albino Rats

Aim: The study investigated the possible risks associated with gastrointestinal ulcer disease by evaluating the biochemical response of three body organs; heart, kidney and liver, in gastric ulcerated rats.
 Methodology: Twenty male wistar albino rats were used in the study. Gastric ulcer was induced in rats with single oral dose of 400 mg/kg body weight (b.w.) aspirin, 80 mg/kg b.w. indomethacin and 5 ml/kg b.w. acidified ethanol (40:60 v/v). Blood samples were collected into heparinized bottle and centrifuged at 4000 rpm for 10 mins to obtain the plasma. Gastric tissue, liver, kidney and heart were also collected.
 Results: Oral administration of 400 mg/kg b.w. aspirin, 80 mg/kg b.w. indomethacin and 5 ml/kg b.w. acidified ethanol caused a remarkable increase in ulcer index. There was observed a significant (p<0.05) reduction in AST and ALT activities in gastric ulceration caused by aspirin (Asp), with no significant (p<0.05) change in total protein (TP) concentration, lactate dehydrogenase and creatine kinase activity. However, there was increase in creatinine and urea concentration. Acidified ethanol and Indomethacin-induced ulcerated rats showed significant (p<0.05) reduction in all other parameters except ALT and lactate dehydrogenase activities which did not show any significant (p<0.05) change. There was also observed a significant (p<0.05) increase in creatine kinase activity in indomethacin-induced ulcerated rats.
 Conclusion: Overall, the result indicates a link between gastric ulcer and organ toxicity. The use of NSAIDs above the therapeutic doses in the treatment of pains and related illness as well as excess consumption of alcohol is shown to negatively impact the stomach and cause serious damage to different body organs of wistar rats.

Effects of Selenium Supplementation in Obese Selenocysteine Lyase Knockout Mouse (P24-022-19)

Abstract Objectives We previously demonstrated that disruption of selenocysteine lyase (Scly) in mice leads to the development of obesity and metabolic syndrome under selenium deficiency, with effects on glucose homeostasis in the liver. It is unknown whether the physiology of skeletal muscle, a key tissue in glucose disposal, is also affected by the disruption of Scly. Here, we investigated the effects of a high-fat diet supplemented with selenium in the muscle physiology of Scly knockout (KO) mice Methods Age-matched homozygous male and female Scly KO and wild type (WT) from C57BL/6 J background were group-caged and fed for three months to a diet containing 45% kcal fat and 0.25 ppm (adequate) or 1 ppm (high) of selenium as sodium selenite. Serum, liver, soleus, and brown adipose tissue (BAT) were collected and assessed for total glutathione peroxidase (GPX) activity and creatine kinase (CK). Total RNA was extracted from soleus and liver, reverse-transcribed, and used in qPCR. Results Following an adequate selenium intake, total GPX activity in serum, mRNA of GPX1 and selenoprotein P in the liver, and mRNA of selenoprotein N in soleus were unchanged of Scly KO compared to WT mice, both in males and females. We observed five-fold increase of CK activity in serum and a one-fold decrease in soleus of Scly KO male mice when compared with WT. Increased CK levels in serum have been related with muscle impairment. Interestingly, when male Scly KO mice were fed a high selenium diet, we observed the opposite effect, with serum CK activity significantly decreased, while in soleus CK activity increased in Scly KO mice. CK activity levels on liver or BAT were unchanged. CK activity in serum was unchanged in female mice, corroborating to the previously described sexual dimorphism for these animals. These results suggest an improvement in Scly KO muscle health after selenium supplementation. Conclusions Adequate selenium levels maintain selenium homeostasis and ensure the maintenance of selenoprotein levels, but high selenium intake may be necessary to preserve muscle health. Funding Sources FAPESP/Brazil, NIDDK-NIH/USA, NIMHD-NIH/USA.

Trimetazidine combined with exercise improves exercise capacity and anti-fatal stress ability through enhancing mitochondrial quality control

AimsTo explore the effects of trimetazidine combined with exercise on EC and anti-fatal stress ability, and illustrate the underlying mechanism. MethodsC57BL/6 mice were randomly assigned to four groups (n = 11 in each group): the control, exercise, trimetazidine and trimetazidine + exercise (TE) groups. Mice were accordingly given saline (ig), Aerobic exercise (AE), trimetazidine (ig), or a combination of trimetazidine (ig) and AE for five weeks. After the intervention, each group was randomly subdivided into rest and exhaustive exercise (EE) subgroups. The mice in the control-EE and TE-EE subgroups underwent fatal stress experiments. EC and anti-fatal stress ability were assessed respectively. Mitochondrial quality control (MQC) in skeletal muscle were measured at the protein level and the organelle level. Key findingsA significantly increased exhaustive swimming time was observed in exercise (39.10 ± 12.58 min vs 14.18 ± 4.37 min), trimetazidine (33.73 ± 8.45 min vs 14.18 ± 4.37 min) and TE groups (73.78 ± 18.95 min vs 14.18 ± 4.37 min) compared with that in the control group, and a synergistic effect was detected (P < 0.05). Fatal stress experiments successfully induced skeletal muscle damage, including increased creatine kinase activity, myofibrosis, and impaired antioxidative enzyme system, all those were significantly alleviated by trimetazidine supplementation combined with AE precondition (P < 0.05). Meanwhile, AE and trimetazidine alone or combined, significantly enhanced the MQC in normal mice by activating mitochondrial biogenesis, dynamics and mitophagy, and that in mice underwent fatal stress stimulus (P < 0.05). SignificanceThis study for the first time found that trimetazidine and AE have synergistic effects on improving EC. Moreover, the combination of both interventions enhances anti-fatal stress ability. Enhancing MQC may be a key mechanism of AE combined with trimetazidine that improves EC and anti-fatal stress ability.

Can exercise-induced muscle damage be related to changes in skin temperature?

Objective: Measurement of skin temperature using infrared thermography has become popular in sports, and has been proposed as an indicator of exercise-induced muscle damage after exercise. However, the relationship between skin temperature and exercise-induced muscle damage is still unclear. Here we set out to investigate the relationship between skin temperature and exercise-induced muscle damage. Approach: Twenty untrained participants completed a protocol of exercise for calf muscles. Before and after exercise blood samples were collected to determine creatine kinase and acetylcholinesterase activity. Thermal images were recorded from the exercised muscles to determine skin temperature. Delayed onset muscle soreness was quantified. Correlations between skin temperature and exercise-induced muscle damage were analyzed considering thermal data, creatine kinase and acetylcholinesterase activity at different time moments. Main results: We found delayed onset muscle soreness and an increased creatine kinase activity 48 h after exercise (P  <  0.01). Skin temperature parameters (average, maximal, amplitude and difference pre- and post-exercise, immediately after and 48 h after) did not correlate with the creatine kinase responses (P  >  0.05). Acetylcholinesterase activity remained stable (P  =  0.59). Significance: We recommend caution when considering changes in skin temperature as dependent on the level of localized and symmetric muscle damage considering calf muscles in untrained participants.

Aldosterone‐producing adrenocortical carcinoma with myxoid differentiation in a cat

A 10-year-old male neutered Persian cat was presented with an abdominal mass and history of weakness. Blood smear examination found marked elliptocytosis, and serum biochemical analysis revealed hypokalemia, hypochloremia, increased creatine kinase activity, and a high aldosterone concentration. Cytologic examination of the mass revealed neoplastic endocrine cells with moderate criteria of malignancy, favoring adrenocortical neoplasia. The adrenal mass was surgically excised and histologically characterized by lobules of mildly pleomorphic, polygonal neoplastic cells with moderate to abundant, occasionally granular, eosinophilic cytoplasm. Lobules were separated by fine fibrovascular trabeculae, and numerous cystic cavities containing amorphous eosinophilic material that stained positive with Alcian blue and periodic acid-Schiff were seen. Neoplastic cells were multifocally positive for cytochrome P450 aldosterone synthase. Based on clinicopathologic and immunohistochemical findings the present case was diagnosed as an aldosterone-producing adrenocortical carcinoma with myxoid differentiation. While this entity has not been reported in cats, myxoid differentiation of adrenocortical carcinomas has been found in other species and can pose a major diagnostic challenge on microscopic examination.