High Creatine Kinase Activity Research Articles

Creatine kinase activity and isoenzymes in lung, colon and liver carcinomas.

We have compared the levels of creatine kinase (CK) activity and the distribution of CK isoenzymes determined by agarose gel electrophoresis in normal colon, liver and lung tissues, and in colon, liver and lung adenocarcinomas, lung squamous cell carcinomas and lung carcinoids. Colon and lung adenocarcinomas, and squamous cell carcinomas presented lower CK activity than the normal tissues and no differences were found between hepatocarcinoma and normal liver tissue. In contrast, lung carcinoids had higher CK activity than normal lung tissue. Type BB-CK was the predominant isoenzyme in normal lung, colon and liver tissues. Type MM isoenzyme was detected in normal lung and type MB-CK was found in normal colon. In most lung tumours the CK isoenzyme electrophoretic pattern did not change. However, no type BB-CK was detected in some hepatocarcinomas, type MM-CK decreased in lung carcinoids and type MB isoenzyme was not observed in colon adenocarcinomas. It is concluded that in most tumours there is a decrease in the expression of type B- and type M-CK subunits, whereas in lung carcinoid the expression of type B-CK activity increases. Thus, the increase in type BB-CK observed in the serum of patients with lung and colon adenocarcinomas is probably due mainly to enhanced enzyme release as a result of tumour cell necrosis.

Serum biochemical values in sled dogs before and after competing in long-distance races

Objective— To measure and compare blood values in sled dogs before and after long-distance racing. Design— Prospective study. Animals— 17 adult sled dogs in the 1991 Iditarod Trail Sled Dog Race and 21 in a simulated sled dog race. Procedure— Blood samples were obtained from 17 dogs 7 days before they began and after they finished (finisher group) or were eliminated from (nonfinisher group) the Iditarod Trail Sled Dog Race. Blood samples were also obtained from 21 dogs before and after a simulated sled dog race. Results— In finisher-group dogs, BUN and uric acid (UA) concentrations were increased after racing; nonfinisher-group dogs had significantly lower postrace BUN and UA concentrations. Significant increases in creatine kinase (CK) and aspartate transferase (AST) activities were detected in all dogs after racing, and postrace values were higher in nonfinisher-group dogs, compared with finisher-group dogs. Mean alkaline phosphate activities were significantly increased after racing in nonfinisher-group dogs only. In dogs that ran the simulated race, postrace values for serum albumin, total protein, calcium, and potassium concentrations, as well as Hct, hemoglobin concentration, and RBC count, were significantly lower than prerace values. Postrace values for alkaline phosphate, alanine transaminase, AST, lactate dehydrogenase, CK, BUN, and UA were significantly higher than prerace values. Clinical Implications— High CK activities are indicative of severe muscle degeneration and, in sled dogs, may represent a degree of muscle breakdown beyond which a dog cannot continue to work. Markedly high CK, and possibly AST, serum activities may be indicators of performance failure in sled dogs competing in long-distance races. (J Am Vet Med Assoc 1997; 211:175–179)

Creatine kinase in trout male germ cells: purification, gene expression, and localization in the testis.

High creatine kinase (CK) activity (16.5 +/- 7.6 IU/mg) is present in trout spermatozoa. In order to partly characterize the CK isozyme predominantly present in sperm and to study the expression of this protein in spermatogenesis, we purified to homogeneity a CK (s-CK) from trout sperm, by nitrogen cavitation followed by two chromatography steps (DEAE-Trisacryl and Blue Sepharose). Specific antisera to 5-CK were developed. A cDNA encoding for a CK named TCK1, and whose transcript shows enhanced testicular expression, was previously isolated from trout testis (Garber et al., 1990: Biochim Biophys Acta 1087:256-258). A CK subunit expressed in vitro by this cDNA cross-reacts with anti-s-CK. A 21-amino-acid residue sequence near the N-terminus of s-CK is identical to the cDNA-derived sequence of TCK1, which is unlike any previously reported CK sequence. Using in situ hybridization, the TCK1 mRNA was detectable in primary and secondary spermatocytes and in early spermatids. Immunohistochemical staining of testis and various organs revealed that s-CK was confined to testis and, in this organ, to late spermatids and spermatozoa. In gill, some cells exhibited a positive signal, but another study rules out the presence of s-CK in this organ (Garber et al., 1990: Biochim Biophys Acta 1087:256-258). These results demonstrate that s-CK/TCK1 is a germ cell-specific protein, the transcription of which starts in meiotic germ cells, while translation starts in late spermatids.

Differential effects of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors on the development of myopathy in young rats.

HMG-CoA reductase inhibitors (statins), cholesterol-lowering drugs that have not been approved for use in children and adolescents, may cause myopathy as a side effect. We compared the effects of three statins (simva-, prava- and lovastatin) in young rats to determine whether skeletal muscle of young animals is more susceptible than that of adults. We also evaluated whether the type of statin (lipophilic versus hydrophilic) determines the degree of muscle damage. Administration via chow of simvastatin (15 mg/kg of body weight/d) and lovastatin (43-55 mg/kg of body weight/d), both lipophilic, caused stunted growth, high creatine kinase (CK) activity in plasma, and severe myopathy. Statin doses that caused damage were much lower for young rats than for adults. Pravastatin (8-55 mg/kg of body weight/d), a hydrophilic drug, caused none of these symptoms. Histologic analysis of hind paw muscles of simvastatin-and lovastatin-treated rats showed abundant signs of damage (hypercontraction, fiber necrosis) in the extensor digitorum longus, correlating with the symptoms noted above. No cellular infiltrates were seen at the onset, pointing to a noninflammatory myopathy. Pravastatin-treated rats never showed signs of myopathy. Impaired DNA synthesis may explain why muscle toxicity is seen at lower doses in young, rapidly developing rats than in adult animals. The differences in muscle damage between the statins may be attributed to differences in lipophilicity and thus in tissue selectivity. Our results can be important when considering drug therapy in young patients with inherited lipoprotein disorders.

Cardiac troponin T in patients with high creatinine concentration but normal creatine kinase activity in serum

Cardiac troponin T in patients with high creatinine concentration but normal creatine kinase activity in serum

In vitro determination of creatine kinase substrate fluxes using 31P-nuclear magnetic resonance

Forward ( k f) and reverse ( k r rate constants and the corresponding flux rates of the creatine kinase catalysed reaction between creatine phosphate (CrP) and adenosine triphosphate (ATP); CrP + ADP ▪ ATP + Cr were measured in vitro at 295 K. Both rate constants were determined using magnetic resonance saturation transfer techniques. To study the dependence of k r and k f on the creatine kinase concentration, the ceatine kinase activity was varied from 2400 to 75 U · ml −1. At equilibrium and high creatine kinase activities, the forward to reverse flux rate ratios are close to 1. A dispersion in the reaction rate constants was observed at activities ≤ 600 U · ml −1. We measured k r > k f for all enzyme activities below 600 U · ml −1. This observation could partially be explained by the presence of ATPase contamination in the enzyme. These findings are relevant for the in vivo studies of creatine kinase activity in the presence of multi-site phosphate exchange in cellular ATP-pools. As mitochondrial creatine kinase is not in equilibrium these results are also of interest in this area.

Influence of mitochondrial creatine kinase on the mitochondrial/extramitochondrial distribution of high energy phosphates in muscle tissue: evidence for a leak in the creatine shuttle.

The influence of mitochondrial creatine kinase on subcellular high energy systems has been investigated using isolated rat heart mitochondria, mitoplasts and intact heart and skeletal muscle tissue. In isolated mitochondria, the creatine kinase is functionally coupled to oxidative phosphorylation at active respiratory chain, so that it catalyses the formation of creatine phosphate against its thermodynamic equilibrium. Therefore the mass action ratio is shifted from the equilibrium ratio to lower values. At inhibited respiration, it is close to the equilibrium value, irrespective of the mechanism of the inhibition. The same results were obtained for mitoplasts under conditions where the mitochondrial creatine kinase is still associated with the inner membrane. In intact tissue increasing amounts of creatine phosphate are found in the mitochondrial compartment when respiration and/or muscle work are increased. It is suggested that at high rates of oxidative phosphorylation creatine phosphate is accumulated in the intermembrane space due to the high activity of mitochondrial creatine kinase and the restricted permeability of reactants into the extramitochondrial space. A certain amount of this creatine phosphate 'leaks' into the mitochondrial matrix. This leak is confirmed in isolated rat heart mitochondria where creatine phosphate is taken up when it is generated by the mitochondrial creatine kinase reaction. At inhibited creatine kinase, external creatine phosphate is not taken up. Likewise, mitoplasts only take up creatine phosphate when creatine kinase is still associated with the inner membrane. Both findings indicate that uptake is dependent on the functional active creatine kinase coupled to oxidative phosphorylation. Creatine phosphate uptake into mitochondria is inhibited with carboxyatractyloside.(ABSTRACT TRUNCATED AT 250 WORDS)

Kinetic evaluation of muscle damage during exercise by calculation of amount of creatine kinase released.

To quantify the extent of muscle alteration during prolonged exercise, the release rate of creatine kinase (CK) from striated muscle was measured in six horses during a rest period (6 h) and during three exercise tests (15, 30, and 60 km) at a constant speed of 200 m/min. CK clearance was measured after intravenous bolus administration (150 U/kg) of a CK solution obtained from horse muscle. The CK steady-state volume of distribution was 0.059 +/- 0.0215 l/kg, the terminal half-life was 123 +/- 28 min, and the plasma clearance was 0.36 +/- 0.10 ml.kg-1 x min-1. After an intramuscular CK administration, the CK systemic availability was 74.1 +/- 21.2% and the half time of absorption was 9.4 +/- 5.7 h, indicating a slow process for CK transit through the lymphatic system. The CK release rate was only significantly increased during the 60-km exercise test. The increase of CK plasma activity was observed after a delay of approximately 5 h and peaked after the end of the race; the estimated CK release rate was 9.92 +/- 2.62 U.kg-1 x h-1 over a mean duration period of 65.8 +/- 15.8 h. With the CK activity of horse striated muscle taken into account, a 60-km race released a quantity of CK corresponding to an equivalent of 18.8 +/- 4.3 g striated muscle. It is concluded that the equivalent amount of damaged muscle may be considered as negligible for a 60-km test and that only very high plasma CK activity levels (at least higher than 10,000 U/l) may provide some evidence of a myolysis.

Muscle histopathology and plasma aspartate aminotransferase, creatine kinase and myoglobin changes with exercise in horses with recurrent exertional rhabdomyolysis.

Six horses with a history of recurrent exertional rhabdomyolysis (RER) (Horses A-F) and 7 control horses performed a submaximal and later a near-maximal treadmill exercise test. Blood samples were obtained before, during and after exercise and muscle biopsies were taken before and after exercise. At rest, plasma aspartate aminotransferase (AST) activities in horses with RER were above 95% confidence intervals for control horses. During submaximal exercise, 3 horses with RER (A, B and C) had much greater increases in plasma AST, creatine kinase (CK) and myoglobin concentrations than did Horses D, E and F and control horses. Clinical signs of muscle stiffness and pain were only obvious in Horse A. During near-maximal exercise, only Horse C showed a substantial increase in CK activity and myoglobin concentrations without any associated clinical signs of rhabdomyolysis. Muscle biopsies from Horses A, B and C contained necrotic type II fibres which, on electron microscopic examination, contained disrupted myofibrils and swollen mitochondria. These results suggest that, in RER, subclinical episodes of muscle fibre necrosis and associated increases in plasma AST, CK and myoglobin occur with exercise more frequently than could be detected clinically. Furthermore, the pattern of increase in muscle enzymes and myoglobin concentrations in the 6 horses with RER suggested that the high plasma AST and CK activities commonly observed at rest in symptom-free Standardbred horses are probably a result of repeated subclinical episodes of rhabdomyolysis after exercise, rather than leakage due to abnormal sarcolemmal permeability.

Haematological and clinicochemical blood profiles in slaughter pigs.

In a cohort study, 40 pig finishing herds were selected: twenty pig herds with a low and twenty pig herds with a high prevalence of several pathological lesions recorded at slaughter in a six-month period before the start of the study. Blood samples were taken from 20 pigs per herd at the end of the finishing period to investigate haematological and clinicochemical profiles. There was only a significant difference in serum albumin concentration between the low and high lesion prevalence groups. There were distinct differences in blood profiles between pig herds, but also between batches of pigs within a herd, housed in different compartments. Differences between castrated males and gilts were also demonstrated with respect to mean values of the blood variables haemoglobin, iron, copper, beta-globulin, eosinophils and segmented neutrophils. However, the differences were not of a biologically important magnitude. The mean values of the blood variables pepsinogen and lymphocytes differed significantly between pig herds from the two participating integration groups. Pigs with a higher albumin concentration, a lower gamma-globulin concentration, a higher copper concentration and a higher creatine kinase activity in serum showed a higher daily weight gain.

Myopathie de l'adulte par déficit en maltase acide. Essai de traitement par régime hyperprotidique

A 21-year old women with rhizomelic muscular deficit and signs of hypercapnia developed acute respiratory failure. Laboratory tests revealed high creatine kinase activity, and electromyograms showed myogenic patterns with a few myotonic discharges. Biopsy of the quadriceps muscle elicited major vacuolar myopathy with glycogen overload. Acid maltase activity was undetectable in muscular tissue. After 7 months on high-protein diet (1540 calories, 37% proteins) there was no clinical or biochemical improvement. The other published cases of acid maltase deficiency treated with high-protein diet are discussed.

Creatine kinase activity in patients with brittle asthma treated with long term subcutaneous terbutaline.

Infused beta 2 agonists have been shown to cause focal myocardial necrosis. Serum creatine kinase activity was compared in 13 patients with brittle asthma currently being treated with subcutaneous terbutaline and an age and sex matched control group of patients with moderate asthma having inhaled treatment only. The median serum total creatine kinase activity for patients receiving subcutaneous terbutaline (211 units/l) was greater than that for the control group (120 units/l). The cardiac specific isoenzyme component of creatine kinase was not raised in either group, and the electrocardiograms and serum aspartate aminotransferase activity were normal. Electromyograms in five patients receiving subcutaneous terbutaline with high creatine kinase activity showed changes consistent with myositis in two, one of whom was subsequently shown to have a metabolic myopathy, which is thought to be long standing. No pathological changes were seen in the myocardium at necropsy in a patient who died from an acute attack of asthma while taking subcutaneous terbutaline. These results suggest that the raised creatine kinase activity seen in patients receiving this treatment is unlikely to be myocardial in origin.

High myocardial mitochondrial creatine kinase activity during ischemia is combined with impaired recovery of contractility after reperfusion

High myocardial mitochondrial creatine kinase activity during ischemia is combined with impaired recovery of contractility after reperfusion

Automated quantification of creatine kinase MB isoenzyme in serum by radial partition immunoassay, with use of the Stratus analyzer

We evaluated the analytical and clinical performances of a new radial partition immunoassay for measuring the mass concentration of creatine kinase (CK)-MB in serum. All pipetting, washes, incubations and data reduction were performed in 8 min by the Stratus (Dade) fluorometric analyzer. Within-assay and between-assay CVs were respectively 5.5% and 8.4% at 21 micrograms/L, and 4.2% and 3.4% at 48 micrograms/L. Assaying serial dilutions of serum samples with high CK-MB concentrations demonstrated excellent linearity. Results of the Stratus technique correlated well (n = 115, r = 0.98) with those of the Tandem-E CKMB II assay. There was no interference from hemolysis, bilirubin, rheumatoid factor, or added CK-MM (up to 3500 U/L); consequently, CK-MB can be determined in undiluted serum, even in the presence of high total CK activity. The mean CK-MB concentration in 105 blood donors was 1.9 (SD 1.3) micrograms/L. For seven myocardial infarction patients who received prompt fibrinolytic therapy, the mean CK-MB concentration was 4.5 (SD 1.8) micrograms/L at admission, and maximum concentrations, 119 (SD 94) micrograms/L, were recorded 16 h later. CK-MB returned to concentrations less than 10 micrograms/L within 72 h.

Energy transport and cell polarity: Relationship of phosphagen kinase activity to sperm function

The energy required for motility of sea urchin sperm is transported from the mitochondrion to the flagellum by a phosphocreatine shuttle involving diffusion of phosphocreatine (PCr) between isozymes of creatine kinase (CrK) localized at the two sites (Tombes and Shapiro, Cell, 41:325, '85; Tombes et al., Biophys. J., 52:75, '87). The present studies demonstrate that high sperm CrK (various echinoderms; sea squirt, bristle worm, salmon) or arginine kinase (molusc, barnacle, moth) activity is seen in several species with sperm of a primitive morphology (mitochondrion at the base of the head, relatively long flagellum). In contrast, CrK activity is 10-100-fold less abundant in sperm of other species (frog, mouse, rooster, rabbit, bull, and human) that either possess a modified morphology (mitochondria that extend along the flagellum) and/or utilize glycolytic metabolism. We interpret these findings as support for the use of phosphagen kinase-dependent energy transport in cells in which the production of adenosine triphosphate (ATP) by the mitochondrion is distant from its utilization, leading to a form of metabolic polarization. Two other cell types, frog photoreceptors and rabbit oviduct cells, whose morphology and function also suggest that they exhibit metabolic polarization, contain relatively high CrK activity. The presence of high phosphagen kinase activity in metabolically polarized gametes and somatic cells further substantiates the role of such enzymes in facilitating energy transport.

Reversible MM-creatine kinase binding to cardiac myofibrils.

Skinned rat papillary muscles and purified preparations of rat cardiac myofibrils were used to study the nature of the interaction of creatine kinase with cardiac myofibrils. High activity of creatine kinase (2 IU/mg protein in fibers and 0.9 IU/mg in purified myofibrils) was due mostly to reversibly bound enzyme. This activity could be removed and rebound. The process of creatine kinase rebinding was characterized by apparent Km value of 0.14 mg/ml (approximately equal to 2 X 10(6) M). Rebinding of creatine kinase to cardiac myofibrils restored the phenomenon of functional compartmentation of adenine nucleotides in myofibrillar space and restored the ability of phosphocreatine to decrease the rigor tension in the presence of MgADP. The physiological experiments with quick length changes showed that rebinding of creatine kinase to skinned papillary muscle also restored Ca sensitivity, increased maximal tension development, decreased stiffness, and restored the tension recovery after quick length changes in muscle under condition of inhibition of endogenous creatine kinase by 1-fluoro-2,4-dinitrobenzene. It is concluded that creatine kinase reversibly bound to cardiac myofibrils is involved in the energy supply for cardiac contraction.

An examination of some factors influencing creatine kinase in the blood of patients with muscular dystrophy

The natural variability of plasma creatine kinase activity has been examined in patients suffering from muscular dystrophy and in normal subjects. The coefficient of variation of the plasma creatine kinase activities was found to be large (approximately 35%) in both patients with Duchenne muscular dystrophy and normal control subjects. A comparison of the plasma activities of creatine kinase with other muscle-derived enzymes suggests that the cause of this variability is changes in the release of enzymes from muscle. Data obtained concerning the effect of physical activity on plasma creatine kinase activity are contradictory, but several young patients with Duchenne muscular dystrophy and a very high creatine kinase activity (greater than 5000 IU/liter) showed a decreased activity following admission to hospital. An estimate of the rate of efflux of certain kinase from muscle has been made, indicating that young ambulant patients with Duchenne muscular dystrophy have a grossly elevated muscle creatine kinase efflux (495.0 +/- 61.3 IU/kg muscle/hr) compared to control subjects (1.4 +/- 0.5 IU/kg muscle/hr).

Effect of the organophosphorus compounds leptophos and TOCP on creatine kinase activity in hens.

The time trend of plasma Creatine Kinase (CK) activity in hens after a single intravenous injection (iv) of orgaonphosphorus compounds (OPs), such as leptophos, TOCP (tri-ortho-cresyl phosphate) or parathion were measured. CK activities in muscles and nervous tissues of hens on day 21 after iv injection of OPs were also measured.In all hens injected with 30mg/kg body weight of leptophos or 40mg/kg body weight of TOCP, the onset of neurotoxic signs was delayed 8 to 10 days after each treatment, while hens injected with 15mg/ kg body weight of leptophos showed no neurotoxic signs. Hens which were injected intravenously with sublethal doses (2mg/kg body weight) or lethal doses (3mg/kg body weight, saved by atropine) of parathion did not show any abnormalities after recovering from acute toxicity.Plasma CK activity in all hens except hens injected with 2mg/kg body weight of parathion rose significantly on the 1st day after OPs injection. A few days later, plasma CK activity recovered to normal levels in hens injected with 15mg/kg body weight of leptophos or 3mg/kg body weight of parathion. However, the high activity levels were prolonged until the 9th to 11th day in hens injected with 30mg/ kg body weight of leptophos or 40mg/kg body weight of TOCP. After developing delayed neurotoxic signs, plasma CK activities rose again in paralitic hens, though ataxic hens maintamed a normal CK level.On the 21st day after OPs injection, CK activities in some parts of the leg muscles and nervous tissues of hens were relatively higher than those of control hens. It is assumed that there was some correlation between the changes of CK activity in muscles and the severity of delayed neurotoxicity in those hens.It is suggested that plasma CK activity is useful for evaluating the severity of delayed neurotoxic damage, and that there are associations between the prolonging of high plasma CK activity in the several days after OPs exposure and the development of delayed neurotoxic symptoms.

CHANGES OF PURINE METABOLISM DURING DIFFERENTIATION OF RAT HEART MYOBLASTS: 138

Rat heart myoblasts (H9c2) were cloned and cultivated in Eagle's minimum essential medium supplemented with 10% FCS. Clones exhibiting high creatine kinase (CK) activity were investigated. Reduction of FCS to 2% for one week resulted in a differentiation of myoblasts towards myocytes. Surface morphology and ultrastructural changes of myoblasts and differentiated myocytes (transverse banding pattern) were investigated by means of electron and light microscopy.Following enzyme activities (nmoles/mg prot./hr) were measured in myoblasts and myocytes:

Creatine kinase of rat heart mitochondria. The demonstration of functional coupling to oxidative phosphorylation in an inner membrane-matrix preparation.

To define more clearly the interactions between mitochondrial creatine kinase and the adenine nucleotide translocase, the outer membrane of rat heart mitochondria was removed by digitonin, producing an inner membrane-matrix (mitoplast) preparation. This mitoplast fracton was well-coupled and contained a high specific activity of mitochondrial creatine kinase. Outer membrane permeabilization was documented by the loss of adenylate kinase, a soluble intermembrane enzyme, and by direct antibody inhibition of mitochondrial creatine kinase activity. With this preparation, we documented four important aspects of functional coupling. Kinetic studies showed that oxidative phosphorylation decreased the value of the ternary enzyme-substrate complex dissociation constant for MgATP from 140 to 16 microM. Two approaches were used to document the adenine nucleotide translocase specificity for ADP generated by mitochondrial creatine kinase. Exogenous pyruvate kinase (20 IU/ml) could not readily phosphorylate ADP produced by creatine kinase, since added pyruvate kinase did not markedly inhibit creatine + ATP-stimulated respiration. Additionally, when ADP was produced by mitochondrial creatine kinase, the inhibition of the translocase required 2 nmol of atractyloside/mg of mitoplast protein, while only 1 nmol/mg was necessary when exogenous ADP was added. Finally, the mass action ratio of the mitochondrial creatine kinase reaction exceeded the apparent equilibrium constant when ATP was supplied to the creatine kinase reaction by oxidative phosphorylation. Overall, these results are consistent with much data from intact rat heart mitochondria, and suggest that the outer membrane plays a minor role in the compartmentation of adenine nucleotides. Furthermore, since the removal of the outer membrane does not alter the unique coupling between oxidative phosphorylation and mitochondrial creatine kinase, we suggest that this cooperation is the result of protein-protein proximity at the inner membrane surface.