Total Creatine Kinase Activity Research Articles

Profiles of creatine kinase isoenzyme compositions in single muscle fibres of different types

Creatine kinase (CK) isoenzyme compositions of different types of single muscle fibres dissected from soleus (SOL) and extensor digitorum longus (EDL) muscles from rats were examined. CK isoenzymes were separated into cytoplasmic (CK-MM, CK-MB, CK-BB) (muscle, brain and hybrid types, respectively) and mitochondrial (m-CK) isoenzymes. Total CK and CK-MM activities showed the highest activities in fast-twitch glycolytic fibres (FG), lower in fast-twitch oxidative glycolytic (FOG) and the lowest in slow-twitch oxidative (SO) fibres. Conversely, the activity of m-CK was highest in SO, lowest in FG and intermediate in FOG fibres. The activity of CK-MB was highest in SO and lower in FOG and FG fibres. However, the activities of total CK and CK isoenzymes in a single muscle fibre type were not distinguishable from those of another type, and the profiles of CK isoenzyme compositions from the same type of single muscle fibres overlapped over a considerable range. The relationships between the four CK isoenzymes activities in single muscle fibres of different types were not similar. These results suggest that CK isoenzymes of single muscle fibres of different types play different roles in intracellular energy metabolism. Therefore, it is supposed that the CK isoenzyme compositions of single muscle fibres are suitable for their contractive and metabolic properties.

Serum Creatine Kinase Activities in Dogs with Dirofilariasis.

Total serum creatine kinase (CK) and its isozyme activities were determined in dogs with dirofilariasis. Before heartworm removal, total CK and isozyme activities in dogs of the mild group were not different from those in dogs of the heartworm-free group. BB activity was higher in dogs of the hemoptysis group. Dogs of the ascites group displayed a mild increase in MM activity. In dogs of the caval syndrome (CS) group, total CK and MM activities were highest among the heartworm-free and heartworm-infected dogs, and MM isozyme accounted for most (75%) of total CK activity. MB and BB activities were also higher. However, there were no significant differences in CK activities between the surviving and non-surviving cases. In dogs with pulmonary heartworm disease (mild and ascites groups), MM activity correlated significantly with the number of heartworms (r = 0.45), hematocrit value (Ht, r = -0.40), serum alanine aminotransferase (ALT, r = 0.42) and lactate dehydrogenase (LDH, r = 0.46) activities, mean pulmonary arterial pressure (r = 0.64) and total pulmonary resistance (r = 0.50). In dogs with CS, MM activity did not correlate with any parameter, but BB activity correlated with the number of heartworms at the right atrium (r = 0.61), Ht (r = -0.53), ALT (r = 0.80), LDH (r = 0.73) and serum urea nitrogen (r = 0.47). At 1 week after heartworm removal, BB and MM activity decreased in dogs of the hemoptysis and ascites groups, respectively. In dogs of the CS group, total CK and MM isozyme activities decreased markedly (P less than 0.01) regardless of their prognosis.

Identification by isoelectric focusing of creatine kinase-MM isoforms in the plasma and striated muscle of pigs

Total creatine kinase (CK) and CK-MM isoforms were determined in plasma and longissimus dorsi muscle extracts from normal pigs. Based on their total CK activity, the pigs were divided into two groups. Pigs of group 1 (n = 16) had a mean plasma total CK of 298 +/- 16 U/L and the distribution of the CK-MM isoforms was 65.7 +/- 2.5% CK-MM3, 18.9 +/- 1.6% CK-MM2 and 15.3 +/- 1.5% CK-MM. In group 2 (n = 18; 826 +/- 75 U/L total CK) four isoforms were observed: 3.1 +/- 0.9% CK-MM, 67.9 +/- 3.0% CK-MM3, 21.5 +/- 2.3% CK-MM2 and 7.5 +/- 1.3% CK-MM. The differences between the two groups of pigs were significant (p less than 0.001) for CK-MM1 and the presence of CK-MM. Four CK-MM isoforms were also detected in longissimus dorsi muscle homogenates: 45.6 +/- 8.1% CK-MM, 32.6 +/- 11.7% CK-MM3, 16.6 +/- 2.3% CK-MM2 and 5.1 +/- 2.8% CK-MM1. The release of CK-MM isoforms from muscle into plasma seems to be unrelated to the concentration of these isoforms in striated muscle.

Creatine kinase-MB activity after implantation of a cardiac pacemaker

Identification of creatine kinase (CK) isoenzyme CK-MB has made an excellent improvement of the diagnosis of myocardial infarction. 1 Cardiac muscle contains relatively large amounts of CK-MB. CK-MB is also present in skeletal muscle. Injury to skeletal muscle may therefore result in elevation of “non-cardiac” CK-MB and cause difficulties in the diagnosis of myocardial infarction. 2,3 Elevation of CK-MB may also be expected in direct cardiac muscle injury, but previous studies reflecting this relation have shown only a slight increase in CK-MB isoenzyme activity. 4 This study evaluates total CK and isoenzyme CK-MB activity in patients undergoing cardiac pacemaker implantation. In this procedure there is injury to cardiac and skeletal muscle.

Morphological, biochemical, and molecular biological characterization of a rat rhabdomyosarcoma cell line during differentiation induction in vitro.

BA-HAN-1C is a clonal rat rhabdomyosarcoma cell line consisting of proliferating mononuclear tumor cells, some of which spontaneously fuse to form terminally differentiated postmitotic myotubelike giant cells. Exposure to retinoic acid resulted in an inhibition of proliferation and a marked increase in cellular differentiation. The number of myotubelike giant cells significantly increased, and about 30% of the mononuclear tumor cells exhibited morphological features of rhabdomyogenic differentiation which were not observed in the mononuclear cells of untreated cultures. Morphological differentiation was paralleled by an increase in total creatine kinase activity as a biochemical marker of differentiation. These effects of retinoic acid were preceded by an increased expression of proto-oncogene raf and transient expression of proto-oncogene fos. The maximum level of fos expression was observed at 15 min and of raf at 12 hr after exposure to retinoic acid. No expression of the proto-oncogenes src, myb, myc, ros, mos, erbA, and erbB was detected.

Morphological, Biochemical, and Molecular Biological Characterization of a Rat Rhabdomyosarcoma Cell Line during Differentiation Induction In Vitro

BA-HAN-1C is a clonal rat rhabdomyosarcoma cell line consisting of proliferating mononuclear tumor cells, some of which spontaneously fuse to form terminally differentiated postmitotic myotubelike giant cells. Exposure to retinoic acid resulted in an inhibition of proliferation and a marked increase in cellular differentiation. The number of myotubelike giant cells significantly increased, and about 30% of the mononuclear tumor cells exhibited morphological features of rhabdomyogenic differentiation which were not observed in the mononuclear cells of untreated cultures. Morphological differentiation was paralleled by an increase in total creatine kinase activity as a biochemical marker of differentiation. These effects of retinoic acid were preceded by an increased expression of proto-oncogene raf and transient expression of proto-oncogene fos. The maximum level of fos expression was observed at 15 min and of raf at 12 hr after exposure to retinoic acid. No expression of the proto-oncogenes src, myb, myc, ros, mos, erbA, and erbB was detected.

Isoenzyme profiles of creatine kinase, lactate dehydrogenase, and aspartate aminotransferase in the diabetic heart: comparison with hereditary and catecholamine cardiomyopathies

The aim was to investigate the redistribution of isoenzymes, clinically important markers of myocardial necrosis, in the diabetic heart and compare it with that investigated in other types of cardiomyopathies. Myocardial isoenzyme activity of creatine kinase (CK), lactate dehydrogenase (LD) and aspartate aminotransferase (AST) was measured in animals with diabetic, hereditary, and catecholamine cardiomyopathies. Diabetic rats (4 and 8 weeks after intravenous streptozotocin, n = 21), Bio 14.6 hamsters (30, 90, 160 and 240 days old, n = 29), and rats injected with isoprenaline (0.25, 0.5 and 1.0 mg.kg-1.d-1 for 3 weeks, n = 20) were used. Controls were age matched intact animals (n = 8-11). Total CK and CK MM activity decreased in all groups. CK MB and BB decreased by 62 and 52% in diabetic rats, but increased by 40 and 33% in Bio hamsters and by 9 and 96% in isoprenaline treated rats. Thus the CK-B subunit decreased by 61% in diabetics and increased by 33 and 38% in Bio and isoprenaline groups, while the CK-M subunit decreased in all groups. Mitochondrial CK decreased in diabetic and isoprenaline groups. Total LD activity increased in diabetics and decreased in Bio. LD-H subunit increased by 21% in diabetics and decreased by 19 and 18% in Bio and isoprenaline groups. Accordingly the proportion of LD-M subunit, an index of anaerobic metabolism, decreased in diabetics and increased in Bio and isoprenaline groups. Changes in CK-M and CK-B subunits and the LD-M proportion in diabetic heart were normalised by insulin. Total AST activity decreased in diabetics because of the reduction in mitochondrial AST. Increased LD-M proportion and CK-B observed in Bio and isoprenaline groups may be a metabolic "compensation" to decreased myocardial perfusion and substrate. Decreased LD-M proportion and CK-B in the diabetic heart was insulin dependent and may indicate either lack of "compensation" to myocardial ischaemia or absence of ischaemia per se. Decreased myocardial CK and CK MB activity possibly causes underestimation of enzymatically assessed infarct size in the diabetic heart.

Diagnosis of perioperative myocardial infarction by considering relationship of postoperative electrocardiogram changes and enzyme increases after coronary bypass operation

We report the results of enzyme determinations in sera from 88 patients, 65 of whom showed inconspicuous reconvalescence, 14 who had myocardial infarction within 24 h (MI 1) after bypass surgery, and nine with myocardial infarction between 24 and 48 h postoperatively (MI 2). We wanted to determine whether the consequent measurement of activities of total creatine kinase (CK), CK isoenzyme MB (CK-MB), lactate dehydrogenase, alpha-hydroxybutyrate dehydrogenase, and aspartate aminotransferase, conducted as a part of routine laboratory diagnostics, provided meaningful information for diagnosing infarcts besides that obtained from the electrocardiogram. The postoperative mean values of the enzyme activities in blood were significantly different among the three groups; however, only a combined evaluation of CK and CK-MB by means of a discriminant analysis allowed the prediction of MI (sensitivity: MI 1 = 98.5%, MI 2 = 95.4%; specificity: MI 1 = 71.4%, MI 2 = 81.8%). CK greater than 600 U/L or CK-MB greater than 45 U/L supports the diagnosis of acute MI.

Erythrocyte osmotic fragility, serum creatine kinase and its isoenzymes related to body weight in porcine malignant hyperthermia

Erythrocyte osmotic fragility, serum creatine kinase (CK) and its isoenzymes (CK-MM, CK-MB and CK-BB) were determined in pigs susceptible (SMH) and resistant (RMH) to malignant hyperthermia at three different body weights (Group 1, 10 to 25 kg; Group 2, 26 to 55 kg; Group 3, 56 to 95 kg). The halothane challenge test was used for determining susceptibility to malignant hyperthermia (MH). Significantly (P less than 0.05) greater erythrocyte haemolysis at saline concentrations of 80 to 100 mM NaCl occurred only between SMH and RMH pigs for body weight group 2. The determination of blood enzyme activities revealed also a significant (P less than 0.001) increase for total CK, CK-MM and CK-BB in SMH and RMH pigs of body weight Group 2. Furthermore in SMH pigs, total CK, CK-MM and CK-MB activities were significantly (P less than 0.05) greater in the animals of body weight Group 2. It was concluded that, in SMH pigs, weighing between 26 and 55 kg, there is a relationship between erythrocyte osmotic fragility, plasma CK, CK isoenzyme activities and muscle development.

Effect of isoenzyme composition on Kodak Ektachem test results for creatine kinase, lactate dehydrogenase, alkaline phosphatase, and amylase

Seventy-four patients' sera containing macroenzymes or other uncommon enzyme forms were analyzed for total creatine kinase, lactate dehydrogenase, alkaline phosphatase, or amylase activity on an Ektachem 700XR analyzer. The Ektachem test results correlated well with activities obtained by means of liquid reagents on RA-1000. For each enzyme tested, linear-regression analysis yielded data comparable with those for sera containing exclusively the more common isoenzymes. Increased concentrations of intestinal ALP, however, resulted in higher activities of total ALP being reported for the Ektachem.

Dantrolene sodium diminishes exercise-induced muscle damage in the rat

We tested the hypothesis that calcium from the sarcoplasmic reticulum contributes to exercise-induced muscle damage. Dantrolene sodium (Dantrium ®) is a muscle relaxant that affects the flux of calcium over the sarcoplasmic membrane. Rats were treated with dantrolene sodium for a week before a 2 h run on a treadmill. We measured the total creatine kinase activity and isoenzyme composition in plasma before and after exercise. We also prepared muscle sections to correlate the biochemical data with morphological evidence of muscle damage. The treated rats showed a marked decrease (34%) in exercise-induced enzyme efflux, caused by a decrease in the muscle specific isoenzyme: instead of a 13-fold increase found in control rats we observed only a 6.5-fold increase in treated animals. The latter showed significantly less muscle damage 48 h later. It is concluded that dantrolene sodium protects the muscle against exercise-induced damage. The possible mechanism of this protection, and its applications in neuromuscular disease are discussed.

Energetic correlates of cardiac failure: Changes in the creatine kinase system in the failing myocardium

To address the hypothesis that impaired ATP synthesis rates caused by changes in the creatine kinase system is an important mechanism underlying cardiac failure, we measured total creatine kinase activity, isoenzyme composition and creatine content in two animal models of hypertrophy with cardiac dysfunction, the spontaneously hypertensive rat in the transition to failure and the creatine-depleted hyperthyroid rat heart challenged by hypoxia. During the transition from stable compensated hypertrophy to failure characterized by decreased functional capacity, we found that total creatine kinase activity and particularly mitochondrial creatine kinase activity decreased. The decrease in functional capacity, the further increase in heart size and the derangements in the creatine kinase system did not occur if these animals were treated for 6 months with the antihypertensive agents, guanethidine or hydralazine. These results suggest that changes in the creatine kinase system occur coordinately with the transition to failure. To assess whether the changes in the creatine system may be causally linked to decreased functional capacity, we used 31P NMR spectroscopy of isolated perfused hearts to define the high energy phosphate content and cardiac performance of creatine-depleted (approximately 50%) hypertrophied hearts challenged by hypoxia. These hearts displayed greater susceptibility to hypoxic injury with regard to both systolic and diastolic function during and following hypoxia. We also measured total creatine kinase activity in right ventricular biopsy specimens from patients with various forms of cardiomyopathy and low ejection fractions, and found a positive correlation between total creatine kinase activity and ejection fraction. Taken together, these results support the hypothesis that decreasing the energy reserve for ATP synthesis renders the heart more susceptible to systolic and diastolic failure.

Diagnostic problem caused by an atypical creatine kinase isoenzyme in a patient with myocardial infarction

This report describes the diagnostic problem caused by an atypical immunoglobulin-bound creatine kinase isoenzyme in a patient who had a myocardial infarction. In the presence of this atypical isoenzyme, creatine kinase isoenzyme electrophoresis was of no help in determining whether myocardial infarction had occurred. A diagnosis of myocardial infarction was confirmed by carrying out lactate dehydrogenase isoenzyme electrophoresis and finding the characteristic increase in LD1/LD2 ratio and by following the total creatine kinase, aspartate aminotransferase and lactate dehydrogenase activities over a 5-day period. Further investigations were carried out which characterized the atypical isoenzyme as an uncommon type: creatine kinase-BB bound to immunoglobulin A lambda.

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.

Alterations in the creatine kinase system in the myocardium of cardiomyopathic hamsters

Immunochemical and biochemical methods were used to assess quantitatively the changes in the heart creatine kinase system in the myopathic Syrian hamsters, line CHF I46. Cardiomyopathy in I75-200 day old animals was characterized by decreased content of mitochondria and lower total creatine kinase activity. In isolated mitochondria only the creatine kinase activity was decreased while cytochromes aa3 content and respiration rate were unchanged. The share of mitochondrial creatine kinase in the total tissue enzyme activity was decreased from 33% to I8% and that of BB form was elevated from 5% in control to 20%, at unchanged relative level of MM. Immunoassay showed decreased amount of the mitochondrial creatine kinase in the tissue and its decreased ratio to cytochromes aa3. The results show altered expression of creatine kinase isoenzymes in cardiomyopathy.

At what level of serum total creatine kinase activity can measurement of serum creatine kinase MB isoenzyme activity be omitted in suspected myocardial infarction?

The purpose of this study was to establish a discriminatory limit for serum total creatine kinase activity (CK activity) below which CK isoenzyme fractionation is unnecessary.We looked at 2610 serum samples from 1077 consecutive patients with suspected acute myocardial infraction (AMI). The CK activity was determined according to the Scandinavian recommended method. Isoenzymes of CK were separated by agarose gel electrophoresis, followed by fluorometric scanning.When the threshold for CK activity was 150 U/l, none of the samples had a creatine kinase MB isoenzyme activity (CK-MB activity) equal to or higher than 30 U/l (the diagnostic level), which has been found to differentiate between patients with AMI and those without AMI. Only 14 patients (1.3% of all patients investigated) had CK-MB activity peaks between 10 U/l (detection limit) and 30 U/l. Of these, AMI was only diagnosed in one.We recommend that CK-MB activity should be measured only when CK activity is higher than 150 U/l. This would make about 50% of all CK-MB measurements unnecessary.

Diabetes decreases creatine kinase enzyme activity and mRNA level in the rat heart.

Several of the adenosinetriphosphatase enzymes that are responsible for cardiac muscle contraction rely on high-energy phosphates supplied by the creatine kinase (CK) system. Experimental diabetes mellitus has been shown to cause a decrease in the maximal contractile performance of the heart. We postulated that the decrease in contractile performance may be explained in part by a decrease in CK enzyme activity. To evaluate this possibility, we determined the level of CK activity and isoenzyme distribution in ventricular homogenates from normal, diabetic, and insulin-treated diabetic rats. We found that total CK activity was decreased by 35% in diabetic hearts and that a 66% reduction in the cardiac-specific MB isoenzyme occurs. Using a cDNA probe for CK-muscle (M) RNA in Northern blot analysis, we determined that a 61.1% decrease in CK-M mRNA occurs in diabetes. Chronic insulin therapy for 1 mo restores CK-M mRNA levels and enzyme activity. In conclusion, diabetes-induced CK enzyme decreases are mediated in part by a lower level of CK-M mRNA that codes for the major CK-M subunit protein. Decreased performance of the CK system may contribute to diabetic cardiomyopathy.

IgA-CK-BB complex with CK-MB electrophoretic mobility can lead to erroneous diagnosis of acute myocardial infarction.

This patient, on admission, presented with a tentative diagnosis of myocardial infarction: the electrocardiogram showed a nonspecific ST-segment and T-wave abnormalities, and total creatine kinase (CK; EC 2.7.3.2) activity was slightly increased (238 U/L). However, a high electrophoretic value for CK-MB (50% of total CK activity) and the electrophoretic pattern of lactate dehydrogenase (EC 1.1.1.27) isoenzymes ruled out myocardial infarction. The isoenzyme migrating as CK-MB was found later to contain no immunologically normal CK-M subunits, and it was bound to IgA. A mixture of the patient's serum and a human serum control containing all CK isoenzymes showed altered electrophoretic mobility only for CK-BB, indicating that the patient's serum contained antibodies to the B unit of CK. Elution from a Sephadex G-200 column showed that the peak at which most of the anodic CK was eluted corresponded to a molecular mass of approximately 200 kDa. Evidently this atypical isoenzyme was an IgA-CK-BB complex. Because this macro CK type 1 can mimic CK-MB, it may therefore be a source of confusion.

Re-evaluation of the diagnostic utility of serum total creatine kinase and creatine kinase-2 in myocardial infarction.

The diagnostic utility of total creatine kinase activity (I), creatine kinase-2 isoenzyme activity (II), and II as a percentage of I, was examined by receiver-operating characteristic curve and likelihood ratio (LR) analyses in 310 persons admitted to the Coronary Care Unit (151 proven cases of myocardial infarction and 159 non-myocardial infarction controls), from whom blood was sampled at 6-h intervals for 48 h after the onset of chest pain. I was ineffective either as a "rule-in" or as a "rule-out" test within the first 6 h of the onset of chest pain; thereafter, it was an effective test. II was the most effective test during the entire 48-h period. III was more effective than I in the first 24-h period, but was less effective than I during the next 24-h period. The decision threshold for high test sensitivities varies with time over the entire 48-h period, but remains constant for high test specificities. It is essential to tabulate the LR(+) and LR(-) values for both test sensitivity and specificity at constant values to determine the utility of each test at each time interval for respectively ruling out or ruling in a diagnosis of myocardial infarction.

Statistical interpretation of concentrations of magnesium, zinc, calcium, potassium, cholesterols, and creatine kinase isoenzymes in men at different stages of ischemic heart disease.

We present a statistical interpretation of plasma (Pl) and (or) erythrocyte (Erc) concentrations of magnesium, zinc, calcium, potassium, and total high-density lipoprotein (HDL) cholesterol, as well as of the activity of total creatine kinase (CK) and its CK-MB isoenzyme, in 26 men with pre-infarction syndrome (PIS) and 34 men with acute myocardial infarction (MI). Discriminant analysis allowed overall comparison of both groups and determination of the most significant variables: CK and Pl-Zn. By non-hierarchical cluster analysis we defined three homogeneous subgroups among MI men, with CK, CK-MB, and Pl-Zn differing significantly between the groups. In PIS men, Pl-Zn was correlated with Pl-Ca, whereas in MI men Pl-Zn was correlated with Pl-Mg. Stepwise regression indicated that Pl-Zn was the most significant regressor of CK in PIS men and of CK-MB in MI men. All these statistical interpretations support a special role of Pl-Zn in diagnosis and perhaps prognosis. After MI, interleukin-1 release may possibly mediate observed hypozincemia via formation of a heart Zn-metallothionein.