CPK Activity Research Articles

Influence of Prerigor Pressurization on Postmortem Beef Muscle Creatine Phosphokinase Activity and Degradation of Creatine Phosphate and Adenosine Triphosphate

ABSTRACTSemimembranosus muscle was excised immediately following slaughter and pressurized at 103.5 MN−2 for 2 min at 35°C. Creatine phosphate (CP), adenosine‐5′‐triphosphate (ATP) and R‐values (a measure of metabolic rate) were determined at 1, 2, 4, and 24 hr postmortem. Creatine phosphokinase activity (CPK) was measured at 1 and 24 hr postmortem. Pressurization accelerated catabolism of both CP and ATP which, in turn, resulted in a higher ATP turnover value than that of the control. R‐values were significantly correlated (r=−0.95) with degradation of ATP and indicate onset of rigor mortis in treated muscle immediately after pressurization. CPK activity was significantly higher (P<0.05) in the pressurized muscle than in the control at 1 hr postmortem. However, this difference did not exist at 24 hr postmortem.

黒毛和種子牛における白筋症の発生例

White muscle disease was observed in 11 calves 31 to 77 days old of the indigenous Japanese Black breed in Shimukappu, Kamikawa, Hokkaido over a period from April to June, 1983. Dysstasia, dysbasia, palpitation, tachypnea and stiffening of the trunk muscle were main clinical signs of the disease. A decrease was seen in α-tocopherol and seleniumconcentration and a remarkable increase in GOT, GPT, LDH (especially LDH4 and LDH5), and CPK activity in the serum. The affected calves showed a dramatic response to the oral administration of vitamin E and antibiotics and allopathic treatment. Of them, eight recovered and three died suddenly.Degeneration of the trunk and cardiac muscles was noticed. The calves and their mothers had mainly been fed hay and silage in which the selenium content was obviously low. It was assumed that the outbreak of white muscle disease might be due to a deficiency in vitamin E and selenium.

The effects of reperfusion of infarct-related coronary artery on serum creatine phosphokinase and left ventricular function.

In 31 patients with transmural myocardial infarction in whom coronary arteriography was performed within 8 hours after the onset of symptoms, we examined (1) the effect of restoration of coronary blood flow on serum CPK time-activity curve and (2) the relationship between cumulative CPKr and the left ventricular function in the chronic phase. We divided 31 patients into 2 groups: Group A consists of 19 patients in whom coronary reperfusion was established. Group B consists of 12 patients whose coronary artery remained occluded. In group A, the time required to reach peak serum CPK activity was significantly shorter than in group B. When comparing CPKr with percent abnormally contracting segment (%ACS) in 2 groups, correlation between CPKr and %ACS was not good, but it revealed linear relation in both group A and B. CPKr divided by %ACS (CPKr/%ACS) was significantly higher in group A than in group B. We conclude that reperfusion of infarct-related coronary artery changes serum CPK time-activity curve resulting in earlier appearance of peak serum CPK and that infarct size cannot be estimated by serum CPK level alone.

Bovine progressive degenerative myeloencephalopathy ('weaver') of Brown Swiss cattle II

An epidemiological survey of this nervous disorder of the Brown Swiss breed in the USA did not implicate any geographical or environmental factors. There was no sex predilection. The number of recorded cases increased from four in 1972-76 to 46 in 1977-81. There were 32 cases in 1982-83. Breeding records from over half the cases indicated a familial pattern involving five sires. Studies on 32 affected purebred Brown Swiss cattle obtained from dairy farms throughout the USA are described. Owners supplied histories and breeding records, and blood typing was carried out to confirm parentage. The criteria used to establish a clinical diagnosis were: onset of bilateral hind leg weakness and ataxia between 5 and 8 months of age; deficient proprioceptive reflexes, normal motor and sensory reflexes, with no other clinically detectable neurological abnormality; and absence of clinically significant skeletal or muscular abnormality. Familial relationship was also taken into account. Laboratory studies included at least one blood count, routine serum chemistry, serum enzyme levels, and analysis of CSF, and selected cattle were screened for toxicological abnormalities. Three sires were incriminated in 48.6% of the cases, and one was incriminated in 31.4%. Laboratory analyses revealed no abnormal haematological parameters. Although increased activities of certain serum enzymes were found, these were not considered specific. CSF samples from two cases showed CPK activity. Toxicological analyses of cholinesterase activity and blood Cu levels were not significant.

Bovine progressive degenerative myeloencephalopathy ('weaver') of Brown Swiss cattle I

An epidemiological survey of this nervous disorder of the Brown Swiss breed in the USA did not implicate any geographical or environmental factors. There was no sex predilection. The number of recorded cases increased from four in 1972-76 to 46 in 1977-81. There were 32 cases in 1982-83. Breeding records from over half the cases indicated a familial pattern involving five sires. Studies on 32 affected purebred Brown Swiss cattle obtained from dairy farms throughout the USA are described. Owners supplied histories and breeding records, and blood typing was carried out to confirm parentage. The criteria used to establish a clinical diagnosis were: onset of bilateral hind leg weakness and ataxia between 5 and 8 months of age; deficient proprioceptive reflexes, normal motor and sensory reflexes, with no other clinically detectable neurological abnormality; and absence of clinically significant skeletal or muscular abnormality. Familial relationship was also taken into account. Laboratory studies included at least one blood count, routine serum chemistry, serum enzyme levels, and analysis of CSF, and selected cattle were screened for toxicological abnormalities. Three sires were incriminated in 48.6% of the cases, and one was incriminated in 31.4%. Laboratory analyses revealed no abnormal haematological parameters. Although increased activities of certain serum enzymes were found, these were not considered specific. CSF samples from two cases showed CPK activity. Toxicological analyses of cholinesterase activity and blood Cu levels were not significant.

The suppression of myogenic functions in heterokaryons formed by fusing chick myocytes to diploid rat fibroblasts

Heterokaryons were formed by fusing differentiated chick skeletal myocytes to fibroblasts derived from skin, lung or heart cultures. The heterokaryons were analyzed for the synthesis of skeletal myosin light chains, acetylcholine receptor, total CPK activity and the ability to spontaneously fuse to form myotubes. Whereas all of the above myogenic functions were expressed in control heterokaryons formed between myocytes and myoblasts, all were extinguished in the crosses between myocytes and fibroblasts. These results confirm that the suppression of myogenic functions previously observed in cell hybrids involving fibroblastoid tumor cells also occurs in heterokaryons isolated using biochemical inhibitors between diploid fibroblasts and chick skeletal myocytes.

In vitro assessment of myocardial function using rabbit working heart: Myocardial protective effect of GIK during ischemic cardiac arrest.

The recent report on the usage of rat working heart for in vitro assessment of myocardial function has prompted us to modify rabbit Langendorff’s heart, which had been frequently used in the field of pharmacology, to the working preparation. A cannula was inserted into the left atrium of rabbit Langen-dorff’s heart. Oxygenated Krebs-Henseleit’s solution entered through the cannula to the left ventricle, from which it was spontaneously ejected through aortic cannula against a hydrostatic pressure of 100 cm H2O. Functional recoveries of aortic flow, aortic pressure, coronary flow and heart rate were observed after ischemic cardiac arrest induced by infusion of cold Young’s solution. Activities of CPK and LDH in the perfusate were also measured. Almost no recovery of aortic flow was obtained after ischemic arrest for 2 hr, while infusion of GIK at every half hour during arrest produced a significant improvement in the recovery of all functions. The most prominent effect was observed in the aortic flow. Infusion of cold Krebs-Henseleit’s solution during arrest was not able to produce such protective effect. Although higher activities of CPK and LDH appeared in the perfusate after reworking, no protective effect of GIK on spilling of these enzymes could be observed.

Delayed-onset muscular soreness and plasma CPK and LDH activities after downhill running

We tested the hypothesis that running down an incline, during which muscles primarily perform eccentric contractions, causes greater delayed-onset muscle soreness and greater increases in plasma enzyme activities than does running on the level, during which muscles perform similar amounts of concentric and eccentric contractions. Subjective sensations of muscular soreness and plasma activities of CPK and LDH were assessed in seven subjects at 0, 24, 48, and 72 h after 45 min of running (one time on the level and a second time down a 10% incline). Following downhill running (57% of VO2max), significant delayed-onset soreness was experienced in gluteal, quadricep, anterior leg, and posterior leg muscles, and plasma CPK (but not LDH) activity was significantly increased (351% at 24 h). In contrast, following level running (78% of VO2max), no statistically significant soreness occurred in any muscle group, and plasma CPK and LDH activities were not elevated. Thus, our results generally support the hypothesis. Secondarily, we investigated whether delayed-onset soreness with downhill running is accompanied by increases in peripheral white blood cell counts suggestive of inflammation. No such association was observed. We suggest that both delayed-onset muscular soreness and plasma enzyme activities are affected by structural changes in muscle tissue resulting from eccentric contractions.

Creatine kinase from the electric organ of Electrophorus electricus (L.)—isozyme analysis

1. 1. Creatine kinase (CPK) isozymes of extracts from the electric organ, dorsal muscle and brain of Electrophorus electricus (L.) were analysed with Cellogel electrophoresis. A single component corresponding to the MB-form was obtained for both electric organ and the dorsal muscle. The BB-form was present in the brain extract. 2. 2. Upon acetone fractionation of the aqueous extract of electric organ, the final fraction was submitted to gel filtration and presented a single peak of CPK activity. 3. 3. Characterization of this fraction by thin-layer gel filtration indicated an apparent molecular weight of 80,000 which corresponds to the enzyme dimeric structure. 4. 4. The implications of this finding with the muscular origin of the electric organ are discussed.

魚類の血液化学検査法に関する基礎的研究 ＶＩ コイ血しょう成分量に及ぼす試料保存時間の影響

As ?? a ?? fundamental step to settle the haemochemical assessment of health or disease in fish, the variation of plasma constituent levels on the storage time or temperature was investigated in carp Cyprinus carpio, of 500-1, 000g in weight. The blood samples were taken by heart puncture using sodium heparin as anticoagulant. After 10min centrifugation at 3, 000rpm immediately after sampling, plasma samples were extracted and divided into 11 or 12 small tubes and stored at 5°C or -20°C. One sample was pool plasma from 2 or 3 fishes. 6 Constituent levels and 7 enzyme activities were measured in plasma samples stored at 5°C or -20°C for 0, 1, 3, 7, 15, and 30 days. And further, 7 enzyne activities were measured in plasma samples stored at -20°C for 3 hours immediately after sampling. Activities fo GPT, CPK and LDH decreased quickly with the lapse fo stored time. GPT activity decreased to about 90% after 24 hours at 5°C, and to about 70% after only 3 hours at -20°C. CPK activity decreased to about 85% after 24 hours at both 5°C and -20°C. Therefore, it is desirable that these levels are mesured immediately after blood sampling. GOT and Al-P activities were stable for 7 days at 5°C, and for 30 days to -20°C. Acid P activity and levels of glucose and uric acid were stable for 15 days at 5°C. LAP activity and levels fo urea-N, total protein, ablumin and inorganic P were stable for 30 days at both 5°C and -20°C.

Retention of sarcoplasmic calcium inhibits development of the phencyclidine-restraint experimental myopathy

The myopathy induced in the rat by the central nervous system stimulant, phencyclidine (PCP), and restraint is characterized by extensive myofibrillar sarcomere disruption in hind limb muscles and massive increases in plasma creatine kinase (CPK) activity. The effects of dantrolene sodium on this myopathy were studied to determine if modulation of calcium release from the sarcoplasmic reticulum could alter the development of the myopathy. Dantrolene prevented both the sarcomere disruption and the increase in plasma CPK activity produced in the PCP-restraint model. The inhibitory effect was not due to a decrease in the locomotor activity produced by PCP. The findings are consistent with a role for excess sarcoplasmic calcium, originating from the sarcoplasmic reticulum, in the development of this myopathy.

31P NMR saturation transfer measurements of the steady state rates of creatine kinase and ATP synthetase in the rat brain

There are two theories concerning the function of phosphocreatine (PCr) in cellular energy metabolism. In one PCr is seen as a reservoir, functioning to maintain constant cytoplasmic ATP concentrations. The alternate theory postulates that PCr acts as an energy shuttle between the site of ATP production in the mitochondria and the site of utilization in the cytoplasm (reviews [ 1,2]). Both theories require that the exchange flux in the creatine kinase (CPK) reaction be considerably greater than the rate of ATP hydrolysis. This is necessary in order to stabilize ATP concentrations during transitions in workload, or during periods when ATP synthesis is impaired, such as ischaemia. Calculations of cytoplasmic phosphorylation potential [3] and measurements of intracellular pH [4] have been based on the assumption that the CPK reaction is near equilibrium. It is now possible to test this assumption in vivo using the technique of 31P NMR saturation transfer; the flux through the CPK reaction was 5 times the steady state rate of ATP turnover in the Langendorff perfused rat heart 151. The total extractable activities of CPK in the rat brain and heart are similar; 500 pmol . g-’ . min-’ in heart [6] and 200 pmol . g-’ min-’ in brain [7], measured at 25’C. The mitochondrial form of the enzyme accounts for 30-40% of the total activity in the heart [l] and 5% in brain [7]. However, mitochondrial CPK activity in both organs is very similar on a per gram mitochondrial protein basis [8]. Despite these similarities, the metabolic response to a decrease in the efficiency of cellular energy transduction,

Immune recognition in the streptococcal carditis of mice: the role of macrophages in the generation of heart-reactive lymphocytes.

The role of immune T cells and of streptococcus-pulsed M phi in the production of cardiac lesions, as well as the ability of M phi to present streptococcal antigens to mouse lymphocytes, was investigated. Cells of mice infected with extracts of group A streptococcus were able to induce the appearance of heart lesions when transferred to syngeneic receptors as well as to transfer DTH reactions to syngeneic heart extracts. Streptococcus-pulsed M phi were also able to induce heart lesions and an increase in the serum CPK activity when injected into syngeneic receptors. This last phenomenon was only observed in mice aged 5 mo or more. Furthermore, it was shown in an in vitro model of T cell proliferation that peritoneal M phi pulsed with group A streptococci are able to induce a specific response to syngeneic cardiac extracts. M phi pulsed with group G streptococcus failed to induce such a response. Those findings are discussed as part of a model for the induction of rheumatic cardiac lesions in which M phi display a central role by selecting antigenic determinants from the pathogenic organism for presentation to immunocompetent cells.

Beta-Adrenergic relaxation and cAMP kinase activation in coronary arterial smooth muscle.

If beta-adrenergic relaxation of smooth muscle is partly mediated by the adenosine 3',5'-cyclic monophosphate (cAMP) system, then beta-stimulation should be correlated to activation of cAMP-dependent protein kinase (cPK). Studies were performed with bovine coronary arterial strips to identify isozymic forms of cPK and to determine if beta-relaxation is correlated to activation of cPK (reflected by elevated ratios of cPK activity without cAMP to cPK activity with cAMP). Both ion exchange chromatography and a new electrophoretic technique revealed two cPK isozymes (types I and II). No change in cPK activity occurred in strips contracted with 30 mM KCl. In contrast, dose- and time-dependent relaxation during beta-stimulation with isoproterenol was highly correlated to parallel increases in cPK activity. Increased cPK activity was inhibited in assays performed with a specific inhibitor of cPK. Both relaxation and activation of cPK were abolished during beta-adrenergic blockade with propranolol. Relaxation by KCl removal or the ionophore R02-2985, unlike beta-mediated relaxation, did not increase cPK activity. These findings show that beta-mediated relaxation of isolated coronary arterial strips specifically activates cPK, and they support the hypothesis that beta-induced relaxation of vascular smooth muscle involves the cAMP system.

Activity of phosphorylcreatine shuttle enzymes in rat cardiac, fast-, and slow-twitch skeletal muscles

Energy producing and energy utilizing reactions in muscle are coherently linked by a phosphorylcreatine shuttle mechanism. Operational components of the shuttle were evaluated in muscles encompassing a wide metabolic and contractile spectrum. Consistent CPK/ATPase ratios suggest homogeneous function of the myofibrillar component of the shuttle. Mitochondrial CPK/ATPase ratios vary due to 4-fold differences in CPK activity with respect to oxidative phosphorylation. The phosphorylcreatine shuttle mechanism, and the mitochondrial component in particular, seems a relevant, physiological regulator of skeletal and cardiac muscle function.

筋ジストロフィー

The “membrane hypothesis” in the pathogenesis of Duchenne muscular dystrophy (DMD) implicates that primary abnormality resides in the membrane system of skeletal muscle and that the abnormality causes progressive breakdown of muscle cells. In DMD, CPK activity in serum is markedly elevated. Histolo gical studies showed that affected muscle contained many “opaque fibers”, or hypercontracted fibers. In these affected fibers, part of plasma membrane was disrupted and xtracellular space tracers invaded muscle cytoplasm. It is postulated that extracellular calcium also gets into sarcoplasm through disintegrated surface membrane and interferes various function of cells.However, the elevation of serum CPK in DMD is explained by the efflux from necrotic muscle cells rather than by the leakage from live cells. Focal loss of plasma membrane might be a reflection of ongoing cell necrosis. Moreover, there is no information about the nature of genetic defect, which is speculated to reside in muscle membrane.The validity of the membrane hypothesis is discussed and other theories on the pathogenesis are also enumerated.

Changes in the FDP (fibrin and fibrinogen degradation products) value under mental and physical stress. I. Changes in FDP and other indices of physio-biochemical effects of stress loaded on volleyball players during training.

It has been known that CA change during physical exercises and associate with dynamic aspects of the coagulation fibrinolysis system of the blood. Therefore, we studied fibrin and fibrinogen degradation products (FDP) under physical exercises together with LDH, CPK, LA, CRE, Na, K and P. 1) College students of Department of Education majoring in sciences of physical education significantly increased activities of CPK and LDH during the training for volleyball games compared with the pre-training values. This increment was remarkable for those who exercised hard, that is, who were loaded with large physical stress. 2) Changes in LA were included in the range of its normal values. Freshmen who had not been trained much and consequently whose adaptability for physical stress was insufficient increased the LA level significantly relative to the pre-training level. 3) Although changes in P were included in the range of its normal values, significant increase was observed by physical stress. Examinations by individual disclosed that a tendency of changes in P was similar to that in CPK and FDP. 4) The tendency of changes in FDP agreed with that in CPK and P and an increase was observed for those who were loaded with large physical stress. It was further observed that especially for those who were loaded with large mental stress together with physical stress the increase in FDP was evident. Therefore, it has been suggested that FDP can be used as an index of biotic effects of not only physical stress but also mental stress.

Pilot study of pirinixil (BR 931) in various forms of hyperlipoproteinemias

The lipid lowering activity of pirinixil (BR 931), a new agent chemically unrelated to clofibrate, was tested in 12 patients (3 M and 9 F), 6 with predominant plasma cholesterol elevation (types IIA and IIB), and 6 with triglyceride elevation (type IV). Two different daily doses were tested (125 mg b.i.d. and 250 mg b.i.d.), according to a cross-over double blind protocol with treatment sequences of 6 weeks each. Both plasma total cholesterol and triglycerides were lowered 14% with the higher drug dosage. The most significant effects were noted in type II hyperlipoproteinemias, all three type IIB patients exhibiting a highly significant cholesterol reduction. Conversely, in type IV hyperlipoproteinemia, a highly significant very low density (VLDL) cholesterol and triglyceride reduction did not correspond to marked changes in triglyceridemia. In some patients, an increase of high density lipoprotein cholesterol was noted. The drug was very well tolerated, with occasional transient rises of CPK activity.

Effects of bucumolol, a beta-adrenergic blocking agent, and its d-isomer on myocardial infarction produced by coronary artery ligation in rats.

Acute ligation of the left coronary artery of rats produced abnormal Q-wave in the electrocardiogram, tachycardia and frequent ventricular fibrillation. Serum CPK level was elevated, reaching a maximum at 3 to 5 hours after ligation and returning to near the pre-ligation level 24 hours later, when CPK activity in the left ventricle markedly decreased. Pretreatment with bucumolol at 2.5 mg/Kg s.c. and 5 mg/Kg s.c. lessened these changes and increased the survival rate in a dose related manner. d-Bucumolol at 5 mg/Kg, on the other hand, increased survival rate primarily by suppressing ventricular fibrillation without any significant effect on other parameters. These results suggest that the membrane stabilizing action does not contribute to protective actions of bucumolol against myocardial ischemia.

Estimation of creatine phosphokinase and hydroxyproline in the developing limb: its use in evaluating the effect of teratogens on myogenesis and chondrogenesis.

Forelimbs of mouse fetuses were examined for tissue-specific, drug-induced alterations in their biochemical composition. The activity of the enzyme creatine phospholinase (CPK; to estimate myogenesis) and the content of hydroxyproline (HP; to estimate chondrogenesis) were compared in homogenates of control and treated mouse-fetus forelimbs on day 14 of gestation. In addition, the content of DNA, RNA, and protein was also measured. Injection of 6-aminonicotinamide (6-AN) (15 mg/kg) on day 10 resulted in an overall growth retardation in day 14 fetuses and all biochemical parameters tested were reduced. The ratio of PH:CPK was unaffected by 6-AN treatment. Retinoic acid (vitamin A acid; 100 mg/kg), administered to pregnant female mice on day 10, produced severe forelimb defects and resulted in a signific 10 resulted in an overall growth retardation in day 14 fetuses and all biochemical parameters tested were reduced. The ratio of PH:CPK was unaffected by 6-AN treatment. Retinoic acid (vitamin A acid; 100 mg/kg), administered to pregnant female mice on day 10, produced severe forelimb defects and resulted in a signific 10 resulted in an overall growth retardation in day 14 fetuses and all biochemical parameters tested were reduced. The ratio of PH:CPK was unaffected by 6-AN treatment. Retinoic acid (vitamin A acid; 100 mg/kg), administered to pregnant female mice on day 10, produced severe forelimb defects and resulted in a significant reduction in day 14 forelimb HP and RNA content, without altering CPK, DNA, or protein; thus, the HP:CPK ratio was decreased. These results indicated that 1) 6-AN nonspecifically retards growth and cyto-differentiation in limbs; 2) retinoic acid inhibits synthesis of collagen and RNA; 3) retinoic acid has a differential effect upon chondrogenic and myogenic tissues of the limb, and 4) the comparison of HP content and CPK activity in tissue homogenates is an acceptable method of evaluating teratogenic compounds for selective effects.