Putative role of ischemic postconditioning in a rat model of limb ischemia and reperfusion: involvement of hypoxia-inducible factor-1? expression

Hypoxia-inducible factor-1α (HIF-1α) is one of the most potent angiogenic growth factors. It improves angiogenesis and tissue perfusion in ischemic skeletal muscle. In the present study, we tested the hypothesis that ischemic postconditioning is effective for salvaging ischemic skeletal muscle resulting from limb ischemia-reperfusion injury, and that the mechanism involves expression of HIF-1α. Wistar rats were randomly divided into three groups (n=36 each): sham-operated (group S), hindlimb ischemia-reperfusion (group IR), and ischemic postconditioning (group IPO). Each group was divided into subgroups (n=6) according to reperfusion time: immediate (0 h, T0), 1 h (T1), 3 h (T3), 6 h (T6), 12 h (T12), and 24 h (T24). In the IPO group, three cycles of 30-s reperfusion and 30-s femoral aortic reocclusion were carried out before reperfusion. At all reperfusion times (T0-T24), serum creatine kinase (CK) and lactate dehydrogenase (LDH) activities, as well as interleukin (IL)-6, IL-10, and tumor necrosis factor-α (TNF-α) concentrations, were measured in rats after they were killed. Histological and immunohistochemical methods were used to assess the skeletal muscle damage and HIF-1α expression in skeletal muscle ischemia. In groups IR and IPO, serum LDH and CK activities and TNF-α, IL-6, and IL-10 concentrations were all significantly increased compared to group S, and HIF-1α expression was up-regulated (P<0.05 or P<0.01). In group IPO, serum LDH and CK activities and TNF-α and IL-6 concentrations were significantly decreased, IL-10 concentration was increased, HlF-1α expression was down-regulated (P<0.05 or P<0.01), and the pathological changes were reduced compared to group IR. The present study suggests that ischemic postconditioning can reduce skeletal muscle damage caused by limb ischemia-reperfusion and that its mechanisms may be related to the involvement of HlF-1α in the limb ischemia-reperfusion injury-triggered inflammatory response.

Role of the phosphocreatine system on energetic homeostasis in skeletal and cardiac muscles / Papel do sistema da fosfocreatina na homeostase energética das musculaturas esquelética e cardíaca

Adenosine triphosphate is the present energy currency in the body, and is used in various cellular and indispensable processes for the maintenance of cell homeostasis. The regeneration mechanisms of adenosine triphosphate, from the product of its hydrolysis – adenosine diphosphate – are therefore necessary. Phosphocreatine is known as its quickest form of regeneration, by means of the enzyme creatine kinase. Thus, the primary function of this system is to act as a temporal energy buffer. Nevertheless, over the years, several other functions were attributed to phosphocreatine. This occurs as various isoforms of creatine kinase isoforms have been identified with a distinct subcellular location and functionally coupled with the sites that generate and use energy, in the mitochondria and cytosol, respectively. The present study discussed the central and complex role that the phosphocreatine system performs in energy homeostasis in muscle cells, as well as its alterations in pathological conditions.

A adenosina trifosfato é a moeda corrente de energia no organismo, sendo utilizada em diversos processos celulares e indispensável para a manutenção da homeostase celular. Mecanismos de regeneração da adenosina trifosfato, a partir de seu produto de hidrólise – a adenosina difosfato – são, dessa forma, necessários. A fosfocreatina é conhecidamente sua fonte mais rápida de regeneração, por meio da enzima creatina quinase. Assim, a principal função desse sistema é atuar como um tampão temporal de energia. Entretanto, ao longo dos anos, diversas outras funções foram atribuídas à fosfocreatina. Isso ocorreu à medida que foram identificadas diversas isoformas da creatina quinase com localização subcelular distinta e acopladas de forma funcional aos sítios geradores e utilizadores de energia, na mitocôndria e citosol, respectivamente. O presente trabalho discutiu o papel central e complexo que o sistema da fosfocreatina desempenha na homeostase energética nas células musculares, bem como suas alterações em quadros patológicos.

Acute and chronic administration of cannabidiol increases mitochondrial complex and creatine kinase activity in the rat brain

Objective: To investigate the effects of cannabidiol (CBD) on mitochondrial complex and creatine kinase (CK) activity in the rat brain using spectrophotometry. Method: Male adult Wistar rats were given intraperitoneal injections of vehicle or CBD (15, 30, or 60 mg/kg) in an acute (single dose) or chronic (once daily for 14 consecutive days) regimen. The activities of mitochondrial complexes and CK were measured in the hippocampus, striatum, and prefrontal cortex. Results: Both acute and chronic injection of CBD increased the activity of the mitochondrial complexes (I, II, II-III, and IV) and CK in the rat brain. Conclusions: Considering that metabolism impairment is certainly involved in the pathophysiology of mood disorders, the modulation of energy metabolism (e.g., by increased mitochondrial complex and CK activity) by CBD could be an important mechanism implicated in the action of CBD. .

Intravenous human umbilical cord blood improves elctrophysiological and metabolic properties in ISO induced myocardial necrosis in rats.

Rats treated with isoproterenol (ISO, 85 mg/kg, sc, twice at an interval of 24 h) showed a significant increase in heart rate, mean arterial blood pressure, pressure rate index, ST elevation on ECG, and a significant increase in the levels of cardiac marker enzymes- lactate dehydrogenase, and creatine kinase in serum and a significant reduction in superoxide dismutase, and catalase and increase in thiobarbituric acid reactive substance activity in heart tissue. Treatment with Human umbilical cord blood (hUCBC; 500 and 1000 µL, iv, via the tail vein; 2 h after the second dose of ISO) significantly restored back to normal levels and showed a lesser degree of cellular infiltration and infarct size in histopathological and planimetry studies respectively. Thus, hUCBC ameliorates cardiotoxic effects of isoproterenol and may be of value in the treatment of myocardial infarction.

Pulsed ultrasound therapy accelerates the recovery of skeletal muscle damage induced by Bothrops jararacussu venom

We studied the effect of pulsed ultrasound therapy (UST) and antibothropic polyvalent antivenom (PAV) on the regeneration of mouse extensor digitorum longus muscle following damage by Bothrops jararacussu venom. Animals (Swiss male and female mice weighing 25.0 ± 5.0 g; 5 animals per group) received a perimuscular injection of venom (1 mg/kg) and treatment with UST was started 1 h later (1 min/day, 3 MHz, 0.3 W/cm², pulsed mode). Three and 28 days after injection, muscles were dissected and processed for light microscopy. The venom caused complete degeneration of muscle fibers. UST alone and combined with PAV (1.0 mL/kg) partially protected these fibers, whereas muscles receiving no treatment showed disorganized fascicules and fibers with reduced diameter. Treatment with UST and PAV decreased the effects of the venom on creatine kinase content and motor activity (approximately 75 and 48%, respectively). Sonication of the venom solution immediately before application decreased the in vivo and ex vivo myotoxic activities (approximately 60 and 50%, respectively). The present data show that UST counteracts some effects of B. jararacussu venom, causing structural and functional improvement of the regenerated muscle after venom injury.

Effect of acute administration of ketamine and imipramine on creatine kinase activity in the brain of rats / Efeito da administração aguda da cetamina e imipramina sobre a atividade da creatina quinase no encéfalo de ratos

OBJECTIVE: Clinical findings suggest that ketamine may be used for the treatment of major depression. The present study aimed to compare behavioral effects and brain Creatine kinase activity in specific brain regions after administration of ketamine and imipramine in rats. METHOD: Rats were acutely given ketamine or imipramine and antidepressant-like activity was assessed by the forced swimming test; Creatine kinase activity was measured in different regions of the brain. RESULTS: The results showed that ketamine (10 and 15mg/kg) and imipramine (20 and 30mg/kg) reduced immobility time when compared to saline group. We also observed that ketamine (10 and 15mg/kg) and imipramine (20 and 30mg/kg) increased Creatine kinase activity in striatum and cerebral cortex. Ketamine at the highest dose (15mg/kg) and imipramine (20 and 30mg/kg) increased Creatine kinase activity in cerebellum and prefrontal cortex. On the other hand, hippocampus was not affected. CONCLUSION: Considering that metabolism impairment is probably involved in the pathophysiology of depressive disorders, the modulation of energy metabolism (like increase in Creatine kinase activity) by antidepressants could be an important mechanism of action of these drugs.

OBJETIVO: Vários achados clínicos sugerem que a cetamina apresenta efeito antidepressivo. O presente estudo tem como objetivo comparar efeitos comportamentais e a atividade da creatina quinase em regiões específicas do encéfalo após a administração de cetamina e imipramina em ratos. MÉTODO: Ratos Wistar receberam uma administração aguda de cetamina ou imipramina e a atividade antidepressiva foi avaliada pelo teste de nado forçado; a atividade da creatina quinase foi medida em diferentes regiões encefálicas. RESULTADOS: Os resultados mostraram que a cetamina (10 e 15mg/kg) e a imipramina (20 e 30mg/kg) diminuíram o tempo de imobilidade quando comparados ao grupo salina. Também foi observado que a cetamina (10 e 15mg/kg) e a imipramina (20 e 30mg/kg) aumentaram a atividade da creatina quinase no estriado e córtex cerebral. A dose mais alta de cetamina (15mg/kg) e a imipramina (20 e 30mg/kg) aumentaram a atividade da creatina quinase no cerebelo e córtex pré-frontal. Por outro lado, o hipocampo não foi alterado. CONCLUSÃO: Considerando que a diminuição no metabolismo provavelmente está envolvida na fisiopatologia da depressão, a modulação do metabolismo energético (como um aumento na atividade da creatina quinase) por antidepressivos pode ser um importante mecanismo de ação destes fármacos.

Effect of very early atorvastatin initiation for acute myocardial infarction on creatine kinase release

It have been suggested that pre-treatment with a statin agent prior to myocardial infarction limits myocardial creatine kinase release, and thus may act to limit myocardial infarct size in humans. To examine the effect of very early statin initiation for AMI, to the extent of myonecrosis as manifested by peak serum creatine kinase levels. Patients with AMI admitted to Al-Kindy teaching hospital cardiac care unit from February 2007 through February 2008, who fulfilled the inclusion criteria cited in the present study, were randomly assigned into two study groups. The statin group patients have received a single oral dose of 40 mg atorvastatin at time of admission and repeated for the next days until discharge, patients not receiving statin serves as controls. Blood samples were obtained upon admission and every 8 h for another three consecutive samples to identify peak creatine kinase levels. Patients who had statin therapy initiated immediately after hospital admission have similar peak creatine kinase concentrations as compared to those not receiving statin therapy [1020 +/- 621 IU/L vs. 911 +/- 591 IU/L,P= 0.332]. Statin initiation in AMI patients fails to show any observable effect on creatine kinase release, which can be attributed to the need of an extended period for the statin agent to achieve the predictable outcome and suggesting the necessity of statin pretreatment in patients at high risk for AMI

Monitoring of cellular enzymes in the serum of electroplating workers at Coimbatore.

Chromium compounds are potent toxic and carcinogenic substances. With respect to toxicity, hepatic and renal toxicity have been reported both in workers and in animals exposed to chromium (VI). Chromium (VI) compounds induces DNA damage in vivo and in cultured cells as well as the cytotoxicity evaluated by the leakage of lactate dehydrogenase. The present study reports the cytotoxicity of chrome platers who are employed from 8 to 25 years in electroplating industries at Coimbatore, Tamilnadu. Blood samples were collected and estimated for glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatine phosphokinase (CPK) and total protein in the serum. The study revealed that there is a significant elevation in the level of LDH, ALP, CPK and transaminases and a decrease in total protein in serum. The results of the study suggests that chromium (VI), a hepatotoxic chemical may perhaps damage the plasma membrane resulting in leakage of enzymes in to the serum of chromeplaters.

Effects of metallothionein on isolated rat heart / 华中科技大学学报（医学）（英德文版）

To investigate the effects of metallothionein (MT) on isolated rat heart, 16 Wistar rats were randomly divided into 2 groups. In control group (group C), distilled water was injected intraperitoneally and 24 h later isolated hearts were perfused with Langendorff and stored at 4 degrees C for 3 h with histidine-tryptophan-ketoglutarate (HTK) solutions, and then isolated hearts were perfused for 2 h by Langendorff. In experimental group (group E), 3.6% ZnSO(4) was injected intraperitoneally, 24 h later isolated hearts were perfused by Langendorff and stored at 4 degrees C for 3 h with HTK solutions, and then the isolated hearts were perfused for 2 h with Langendorff. MT content, the recovery of hemodynamics, myocardial water content (MWC), lactate dehydrogenase (LDH) and creatine kinase (CK) leakage, adenosine triphosphate (ATP) and malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, myocardial cell Ca(2+) content, Ca(2+)-ATPase activity of mitochondria ([Ca(2+)-ATPase](m)) and its Ca(2+) content ([Ca(2+)](m)), synthesizing ATP activity of mitochondria ([ATP](m)), and the ultrastructure of cells were examined. There were a significant increase in group E in hemodynamic recovery, ATP content, SOD activity, [Ca(2+)-ATPase](m) activity, [ATP](m) activity, and substantial reduction in MWC, LDH and CK leakage, MDA content, myocardial cell Ca(2+) content, [Ca(2+)](m) content, and the ultrastructural injury were obviously milder than that of group C. This study demonstrated that MT has protective effects on isolated rat heart.

Avaliação clínica e da função muscular em pacientes com hipotireoidismo subclínico / Clinical and muscular evaluation in patients with subclinical hypothyroidism

Alguns sintomas e sinais de hipotireoidismo, bem como alterações laboratoriais, podem estar presentes no hipotireoidismo subclínico (HS). Este trabalho avalia a prevalência de sintomas e sinais de hipotireoidismo e alterações músculo-esqueléticas em pacientes com HS (n= 57) comparado a um grupo controle sem disfunção tireoideana (n= 37). Baseado na presença de sintomas e sinais de hipotireoidismo, os participantes receberam pontuação específica (escore clínico). A força muscular foi aferida pelo teste muscular manual e por dinamômetro de cadeira e a força inspiratória por manovacuômetro. Os níveis de hormônios tireoideanos e enzimas musculares foram dosados. O grupo HS apresentou escore mais elevado (p< 0,01), maior freqüência das queixas de mialgia e fraqueza (p< 0,05) e redução de força das cinturas escapular e pélvica (p< 0,05). Os níveis médios de T4 livre foram menores no HS (p< 0,001). Os resultados sugerem que a presença de sintomas e sinais de disfunção tireoideana e níveis de T4 livre na faixa inferior da normalidade no HS possam estar relacionados e devem ser valorizados na decisão de iniciar LT4.

Evaluation of enzymes in serum and cerebrospinal fluid in cases of stroke.

Levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and creatine kinase (CK) were estimated in serum and cerebrospinal fluid of 25 patients of stroke, and were correlated with severity of disease. 21 (84%) patients had ischemic stroke and four (16%) had hemorrhagic stroke. Serum and CSF AST levels were significantly elevated in the study group. The rise in CSF AST was more in the hemorrhagic subtype than in the ischemic subtype. Serum ALT and CSF LDH levels were also significantly elevated in patients with ischemic stroke. None of the enzyme levels were related to the severity of disease as assessed by the Glasgow coma scale.

Serum muscle enzymes, muscle pathology and clinical muscle weakness: correlation in Thai patients with polymyositis/dermatomyositis.

The clinical correlation between serum muscle enzymes, muscle pathology and muscle weakness was studied in 100 Thai patients (22 males and 78 females) with polymyositis (PM) and dermatomyositis (DM). Their mean +/- SD age and duration of disease were 45.0 +/- 13.9 years and 6.3 + 13.4 months, respectively. There was idiopathic PM in 37 cases, idiopathic DM in 13, PM/DM associated with malignancy in 5 and PM associated with connective tissue disease in 45. Serum muscle enzymes including creatine phosphokinase, lactate dehydrogenase and aspartate aminotransferase were elevated in 87 per cent, 92 per cent, and 82 per cent of cases, respectively. Abnormal electromyographic findings that were compatible with inflammatory myopathy were found in 76 per cent of cases. Seventy-seven per cent had an abnormal muscle biopsy that was consistent with polymyositis. There was a significant correlation between serum muscle enzymes and muscle pathology (p < 0.01). The degree of muscle weakness correlated better with the degree of muscle destruction (p = 0.01) than the degree of muscle inflammation (p = 0.03). The erythrocyte sedimentation rate showed no correlation with serum muscle enzymes, muscle pathology or muscle weakness.

New cardiac markers: selecting the best tools for reaching an accurate diagnosis and determining the patient's prognosis.

Many new cardiac markers have evolved over the past one decade. The role of existing cardiac markers is being redefined. Cardiac markers of late have become not only an important diagnostic tool, but also their prognostic significance is slowly being recognised. They have also become a deciding factor on the need of instituting aggressive management protocols like percutaneous coronary interventions. This article takes a latest updated look on these aspects of serum cardiac markers.

Influência do pré-condicionamento isquêmico na proteçäo miocárdica em revascularizaçäo do miocárdio com pinçamento intermitente da aorta / Influence of the ischemic preconditioning in the myocardial protection in myocardial revascularization with intermittent aortic cross-clamping

Objetivo: Este estudo testa a hipótese de que curtos períodos de isquemia podem aumentar a proteçäo obtida pelo pinçamento intermitente da aorta. Métodos: No grupo controle (18), a operaçäo foi realizada com hipotermia sistêmica a 32 ºC com pinçamento intermitente da aorta e uso de circulaçäo extracorpórea (CEC). No segundo grupo, denominado de pré-condicionamento (17), foram acrescidos dois pinçamentos de 3 minutos da aorta com intervalo de 2 minutos de reperfusäo entre eles, previamente ao pinçamento intermitente da forma convencional. CK-MB, troponina I, adenosina e lactato foram obtidos do seio ocoronário no início da circulaçäo extracorpórea (1), ao final da segunda anastomose (2) e ao final da CEC (3). Resultados: Os níveis de CK-MB e troponina I apresentaram uma leve tendência a aumentar ao final da CEC no grupo controle, enquanto os de adenosina e lactato näo apresentaram diferença. (Ver tabela). Conclusäo: Concluímos que o pré-condicionamento isquêmico näo promoveu melhora significante na proteçäo miocárdica

Intoxicaçäo experimental por monensina em equinos / Experimental monensin poisoning in horses

Sete eqüinos foram tratados experimentalmente com monensina sódica. Dois desses animais receberem 3-4 kg/eqüino/dia de uma ração comercial sabidamente implicada em surtos naturais da intoxicação por monensina em eqüinos e que continha 180 ppm±20 da droga. Um eqüino recebeu uma única dose de 5 mg/kg e um outro recebeu 4 doses diárias de 1 mg/kg de monensina sódica originária de um premix. Esses quatro eqüinos morreram ou foram sacrificados in extremis, 3-8 dias após o início da administração da droga. Um quinto eqüino recebeu dose única de 5 mg/kg de monensina, ficou levemente doente e se recuperou. Dois eqüinos não desenvolveram sinais da intoxicação. Um desses eqüinos tinha recebido 40 doses diárias de 0,5 mg/kg de monensina e o outro recebeu 3 kg/dia de uma ração da mesma marca que a usada nas fazendas onde surtos de intoxicação por monensina foram detectados (mas de uma outra partida, mais tarde determinada como contendo menos de 5 ppm de monensina). O aparecimento dos sinais clínicos ocorreu de 2 a 5 dias após a administração da droga e a duração do quadro clínico variou de 24 a 76 horas. Os sinais clínicos incluíam taquicardia, arritmia, gemidos, incoordenação, sudorese, decúbito esternal, decúbito lateral, pedaleios e morte. Em cinco dos eqüinos intoxicados observaram-se marcadas elevações da atividade plasmática de creatina fosfoquinase e, em um eqüino, houve leve aumento da atividade plasmática de aspartato aminotransferase. Os principais achados de necropsia consistiram em áreas brancas ou amarelas, focais ou focalmente extensas e bilateralmente simétricas nos músculos esqueléticos...

Lipid peroxidation and antioxidant enzymes in isoproterenol induced oxidative stress in rat erythrocytes.

Isoproterenol, upon oxidation, produces quinones which react with oxygen to produce superoxide anions (O2.-) and H2O2. In the present study, isoproterenol was administered to rats in two doses so as to evaluate its beta adrenergic and toxicological action in terms of lipid peroxidation (LPO) and antioxidant enzymes in erythrocytes. Isoproterenol (30 mg/100 g body wt.) was administered to rats and the animals were followed up to 7 days after administration. Some of these animals were treated with a second dose of isoproterenol 24 h after the first dose and the animals were followed up to 12 h. The result showed increased lipid peroxidation (LPO) and superoxide dismutase (SOD) activity in erythrocytes in response to isoproterenol. Catalase (CAT) activity in erythrocytes decreased with isoproterenol between day 2-7 as compared to control. The second injection of isoproterenol showed increased CAT activity in erythrocytes which decreased at 12 h as compared to control. The erythrocyte GSH content and glutathione-S-transferase (GST) activity decreased with isoproterenol treatment as compared to control. However, erythrocyte GSH content as well as GST activity both recovered towards control with time. Elevated serum lactate dehydrogenase (LDH), creatine phosphokinase (CPK) and glutamate oxaloacetate transaminase (GOT) activity was observed after isoproterenol treatment. The results show increased LPO and altered antioxidant system in erythrocytes in response to isoproterenol induced oxidative stress.

Efecto protector del dantrolene en las lesiones miocárdicas producidas por la reperfusión post-isquémica / Protective effect of dantrolene in post-ischemic reperfusion myocardial damage

Este estudio muestra que la hidantoína, dantrolene, protege al miocardio de las lesiones ocasionadas por la reperfusión post-isquémica. Este efecto, fue analizado después de ocluir durante 5 minutos la arteria coronaria izquierda de corazones de ratas Wistar. Los resultados evidencian que el dantrolene, a dosis de 1 mg/Kg, es efectivo para proteger al miocardio de las arritmias ventriculares y la disfunción contráctil ocasionadas por la isquemia-reperfusión. Además, disminuye la liberación de las enzimas creatina cinasa y lactato deshidrogenasa al plasma y protege al miocardio del daño estructural. Proponemos que esta acción protectora, puede deberse a un efecto bloqueador del canal liberador de calcio (canal de rianodina) del retículo sarcoplásmico, disminuyendo de este modo la sobrecarga de calcio sarcoplasmico, característico de la lesión por reperfusión

Correlation between creatine kinase activity, lipid-peroxidation and water test in male infertility.

New approaches need to be pursued towards the assessment of sperm quality using biochemical markers. In order to help develop a good biochemical marker to assess sperm-membrane integrity, the enzyme creatine kinase (CK) was studied in semen of normal, oligospermic and azoospermic samples and correlated with sperm concentration, lipid-peroxidation (LP) and water test. Presence of isoforms of creatine kinase (CK-MB) was also seen. An inverse correlation was observed between CK activity and sperm concentration (p<0.001). Water test was seen to be inversely correlated with CK activity (p<0. 001). Lipid peroxidation showed positive correlation with CK activity (p<0.001). A significant correlation between loss of sperm function meditated by induction of peroxidative damage to sperm plasma membrane is indicated. Enzymes like CK can serve as good biochemical marker along with lipid peroxidation to confirm loss of sperm membrane integrity. The water test can be used as a preliminary screening test for sperm membrane integrity.

Effects of vanadate, insulin and fenugreek (Trigonella foenum graecum) on creatine kinase levels in tissues of diabetic rat.

The in vivo effects of insulin, and other insulino mimetic agents like vanadate and fenugreek (T. foenum graecum) were followed on the changes in the activities of creatine kinase in heart, skeletal muscle and liver of experimental diabetic rats. As compared to control rats, creatine kinase activities were found to decrease significantly in the tissues during experimental diabetes. All the antidiabetic compounds used namely, insulin, vanadate and Fenugreek seed powder normalised the decreased activities to almost control values. The effects of insulin and vanadate were comparable in restoring normoglycemia and the creatine kinase activities.