Creatine Kinase BB Activity Research Articles

Oxidative Stress and Serum Creatine Kinase BB Levels can Help Mark Severity and Stage of Brain Tumor

The exact cause of brain tumor is still unknown, and it is a multi-factorial disease. Brain tumor can originate from brain cells or secondary from other organs via metastasis. Generally, tumor cells show aberrant metabolism of free radicals. The current study aims at evaluating creatine kinase BB (CK-BB) and total oxidative stress (TOS) levels among brain tumor patients for diagnosis, prognosis, and treatment strategy. Hospital-based comparative cross-sectional study was conducted on a total of 90 participants (50 brain tumor patients and 40 healthy controls) from April 2018 to October 2018. Venous blood samples were collected for TOSand CK-BB activity measurement. Purposive sampling technique was implemented to select study participants in the hospital. Catalytic activity of CK-BB and TOS were significantly increased in the serum of brain tumor patients as compared to controls. TOS levels in patients with malignant brain tumor were significantly higher than those with benign tumor, but CK-BB levels were not significantly different between benign and malignant types of brain tumor. Both serum levels of TOS and CK-BB were significantly associated with brain tumor. The CK-BB activity was significantly associated with brain tumor and can be used as markers for diagnosis. Participants with increased TOS had significantly higher risk of brain tumor development compared to healthy controls. In conclusion, oxidative stress status and CK-BB level may be used as possible brain tumor markers.

The effects of estrogen receptors α‐ and β‐specific agonists and antagonists on cell proliferation and energy metabolism in human bone cell line

In cultured human osteoblasts estradiol-17β (E2) modulated DNA synthesis, the specific activity of creatine kinase BB (CK), 12 and 15 lipoxygenase (LO) mRNA expression and formation of 12- and 15-hydroxyeicosatetraenoic acid (HETE). We now investigate the response of human bone cell line (SaOS2) to phytoestrogens and estrogen receptors (ER)-specific agonists and antagonists. Treatment of SaSO2 with E2, 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN; ERβ-specific agonist), 4,4',4″-[4-propyl-(1H)-pyrazol-1,3,5-triyl] tris-phenol (PPT; ERα-specific agonist), biochainin A (BA), daidzein (D), genistein (G) and raloxifene (Ral) showed increased DNA synthesis and CK. Ral inhibited completely all stimulations except DPN and to some extent D. The ERα-specific antagonist methyl-piperidino-pyrazole (MPP) and the ERβ-specific antagonist 4-[2-phenyl-5,7-bis (tri-fluoro-methyl) pyrazolo [1,5-a]pyrimidin-3-yl] phenol (PTHPP) inhibited DNA synthesis, CK and reactive oxygen species (ROS) formation induced by estrogens according to their receptors affinity. The LO inhibitor baicaleine inhibited only E2, DPN and G's effects. E2 and Ral unlike all other compounds had no effect on ERα mRNA expression, while ERβ mRNA expression was stimulated by all compounds. All compounds modulated the expression of 12LO and 15LO mRNA, except E2, PPT and Ral for 12LO, and 12- and 15-HETE productions and stimulated ROS formation which was inhibited by NADPH oxidase inhibitors diphenyleneiodonium chloride (DPI) and N-acetyl cysteine and the estrogen inhibitor ICI. DPI did not affect hormonal-induced DNA and CK. In conclusion, we provide evidence for the separation of mediation via ERα and ERβ pathways in the effects of estrogenic compounds on osteoblasts, but the role of LO/HETE/ROS is unclear.

Evaluation of IGL-1 preservation solution using an orthotopic liver transplantation model

To compare, in a pig liver transplantation model, the protective effect of UW with that of IGL-1, a high-sodium preservation solution containing polyethylene glycol (PEG) as an oncotic supply. All livers were harvested and grafted orthotopically according to standard techniques. The livers were washed out and preserved for 7 h in IGL-1 (n = 6) or in UW solution (n = 7) at 4 degree centigrade. In a sham group (n = 4), the livers underwent a 60-min warm ischemia at 37 degree centigrade. The hepatocellular injury was assessed in organ preservation solution washed out from the graft at the end of ischemic storage (before revascularization), and in serum 2 h after reperfusion and daily for up to 6 d. Livers preserved in IGL-1 solution released markedly less AST than that preserved in the UW solution before and after revascularization (P < 0.05). Besides, the activity of creatine kinase-BB, a marker of sinusoidal lining cells injury, was higher in the UW group than in the IGL-1 group (P < 0.05). Histological results showed less necrotic regions in livers preserved in IGL-1 solution; however, no difference was observed for inflammation. IGL-1 liquid effectively protects parenchymal and non-parenchymal cells against preservation-reperfusion injuries.

Increased creatine kinase BB activity and CKB mRNA expression in patients with hematologic disorders: Relation to methylation status of the CKB promoter

Background We encountered 2 patients with increased activities of the creatine kinase (CK)-BB isoenzyme in their sera. Here we examined the relation among CK-BB activity, expression of CKB mRNA in peripheral blood, and hypermethylation of the CKB. Methods The 2 patients and other 26 patients with hematologic malignancies, and some cancer cell lines were subjected to measurement of serum CK activity, CK isoenzyme analysis, CKB mRNA expression analysis by RT–PCR, and methylation analysis of the CKB promoter region. Results CK-BB activity and proportion of leukemia blasts were correlated in the 2 patients. CKB mRNA was increased in peripheral blood during an increase in leukemia blast numbers. In contrast, none of the other 26 patients showed CK-BB activity or expression of CKB mRNA. In all of the patients with hematologic disorders, the analyzed region of CKB promoter was mostly unmethylated. However, some of cancer cell lines showed the methylated pattern. CKB mRNA was expressed at higher levels in cells with an unmethylated CKB promoter than in cells with a methylated promoter. Conclusions Expression of CKB mRNA and CK-B sometimes occurred in blastic transformation of the hematopoietic system. A relation between CKB mRNA expression and methylation of the CKB promoter was suggested.

Increased protein oxidation and decreased creatine kinase BB expression and activity after spinal cord contusion injury.

Traumatic injury to the spinal cord triggers several secondary effects, including oxidative stress and compromised energy metabolism, which play a major role in biochemical and pathological changes in spinal cord tissue. Free radical generation and lipid peroxidation have been shown to be early events subsequent to spinal cord injury. In the present study, we demonstrated that protein oxidation increases in rat spinal cord tissue after experimental injury. As early as h after injury, the level of protein carbonyls at the injury epicenter was significantly higher than in control (169%, p < 0.05) and increased gradually over the next 4 weeks to 1260% of control level. Both caudal and rostral parts of the injured spinal cord demonstrated a mild increase of protein carbonyls by 4 weeks postinjury (135-138%, p < 0.05). Immunocytochemical analysis of protein carbonyls in the spinal cord cross-sections showed increased protein carbonyl immunoreactivity in the epicenter section compared to rostral and caudal sections of the same animal or control laminectomy animals. Increased protein carbonyl formation in damaged spinal cord tissue was associated with changes in activity and expression of an oxidative sensitive enzyme, creatine kinase BB, which plays an important role in the maintenance of ATP level in the CNS tissue. Damage to CK function in the CNS may severely aggravate the impairment of energy metabolism. The results of our study indicate that events associated with oxidative damage are triggered immediately after spinal cord trauma but continue to occur over the subsequent 4 weeks. These results suggest that antioxidant therapeutic strategies may be beneficial to lessen the consequences of the injury and potentially improve the restoration of neurological function.

A non-calcemic analog of 1α,25 dihydroxy vitamin D 3 (JKF) upregulates the induction of creatine kinase B by 17β estradiol in osteoblast-like ROS 17/2.8 cells and in rat diaphysis

We have reported that multiple treatments with so-called ‘non-hypercalcemic’ analogs of 1α,25(OH) 2 vitamin D 3 (1,25(OH) 2D 3) stimulate the specific activity of creatine kinase BB (CK) in ROS 17/2.8 osteoblast-like cells, and that pretreatment with these analogs upregulates responsiveness and sensitivity to 17β estradiol (E 2) for the induction of CK. However, since the analogs showed toxicity in vivo, we have now studied the action of a demonstrably non-calcemic hybrid analog of vitamin D in ROS 17/2.8 cells, and prepubertal rats. The analog JKF was designed to separate its calcemic activity from other biological activities by combining a calcemic-lowering 1-hydroxymethyl group with a potentiating C, D-ring side chain modification including 24 difluoronation. Treatment with 1 pM JKF alone significantly stimulated CK specific activity at 4 h by 30±10%. However after three daily pretreatments, JKF upregulated the extent of induction by 30 nM E 2 by 33% at 1 pM and by 97% at 1 nM; the E 2 dose needed for a significant stimulation of CK activity was lowered to 30 pM. The action of the SERMS tamoxifen, tamoxifen methiodide and raloxifene, at 3 μM, was also upregulated by three daily pretreatments with 1 nM JKF; unexpectedly, this pretreatment prevented the inhibition of E 2 stimulation by the SERMS. Upregulation of E 2 action by 1 nM JKF was inhibited by 1 nM ZK159222, an inhibitor of the nuclear action of 1,25(OH) 2D 3. In vivo, three daily injections of 0.05 ng/g body weight of JKF augmented the response of prepubertal female rat diaphysis and epiphysis to E 2. Therefore, demonstrably non-calcemic analogs of 1,25(OH) 2D 3 may have potential for use in combination with estrogens or SERMS in the prevention and/or treatment of metabolic bone diseases such as postmenopausal osteoporosis.

Oxidative modification of creatine kinase BB in Alzheimer's disease brain.

Creatine kinase (CK) BB, a member of the CK gene family, is a predominantly cytosolic CK isoform in the brain and plays a key role in regulation of the ATP level in neural cells. CK BB levels are reduced in brain regions affected by neurodegeneration in Alzheimer's disease (AD), Pick's disease, and Lewy body dementia, and this reduction is not a result of decreased mRNA levels. This study demonstrates that posttranslational modification of CK BB plays a role in the decrease of CK activity in AD brain. The specific CK BB activity and protein carbonyl content were determined in brain extracts of six AD and six age-matched control subjects. CK BB activity per microgram of immunoreactive CK BB protein was lower in AD than in control brain extracts, indicating the presence of inactive CK BB molecules. The analysis of specific protein carbonyl levels in CK BB, performed by two-dimensional fingerprinting of oxidatively modified proteins, identified CK BB as one of the targets of protein oxidation in the AD brain. The increase of protein carbonyl content in CK BB provides evidence that oxidative posttranslational modification of CK BB plays a role in the loss of CK BB activity in AD.

Vitamin D analogs modulate the action of gonadal steroids in human vascular cells in vitro

We have previously reported that estradiol (E 2) and dihydrotestosterone (DHT) regulate cell growth in human umbilical arterial smooth muscle cells (SMC) and in an endothelial cell line (E304). In SMC both gonadal steroids stimulated DNA synthesis at low concentrations and suppressed 3[H] thymidine incorporation at high concentrations, whereas in E304 cells E 2 and DHT dose dependently enhanced DNA synthesis. In both cell types gonadal steroids also induced the specific activity of creatine kinase BB (CK). Previous evidence suggets that the in vitro and in vivo CK responses to gonadal steroids in bone cells are upregulated by pretreatment with vitamin D analogs due to increased level of cellular estrogen receptors (ER). Here we analyzed the interaction of the vitamin D analogs hexafluorovitamin D (FL), JK-1624 F 2-2 (JKF), and CB 1093 (CB) with gonadal steroids in regulating DNA synthesis and CK activity in human vascular cells in vitro. In E304 cells, daily treatment with FL, JKF, or CB (1 nmol/L for 3 days) increased DNA synthesis by 110 ± 11%, 65 ± 16%, and 88 ± 23% respectively. In contrast, the same analogs inhibited 3[H] thymidine incorporation by 52 ± 21%, 46 ± 19%, and 50 ± 10%, respectively, in SMC. In both cell types all three analogs increased CK by 25% to 75% and amplified the CK response to E 2 and to DHT by twofold to threefold. In E304 cells the vitamin D analogs also increased DNA response to gonadal steroids from 50% to 60% to 200% to 280%. In SMC these analogs did not modify the DNA synthetic response to a low E 2 concentration, but prevented the suppression of DNA synthesis exerted by high concentrations of E 2 and DHT. Vitamin D inhibitors known to block cellular calcium mobilization, had no effect on the proliferative activity induced by vitamin D analogs. However, the inhibitor of the nuclear effects of vitamin D, ZK 159222, blocked the stimulatory effects of CB on DNA synthesis in E304 cells. Finally, both 1,25(OH) 2 D 3, and JKF decreased the expression of ERβ proteins in SMC and increased the ERα isoform in E304 cells by 40% to 75%. The results indicate that vascular cells are targets for both vitamin D and gonadal steroid action and suggest a possible interaction between these hormones in the regulation of cell proliferation via modulation of vascular ER or interaction with proteins associated with ER.

Somatosensory potentials, CSF creatine kinase BB activity, and awakening after cardiac arrest.

To examine the utility of somatosensory evoked potential (SEP) peaks and CSF creatine kinase BB isoenzyme activity (CKBB) in predicting nonawakening from coma due to cardiac arrest. Accurate predictors of neurologic outcome in patients comatose after cardiac arrest are needed to improve medical decision making. A total of 72 comatose patients had bilateral median SEPs, and of these, 52 had CSF and CKBB. Awakening was defined as following commands or having comprehensible speech. Both short (N1) and long (N3) latency SEP peaks were analyzed. Nonparametric analyses were used. For patients who had both tests, CKBB > or = 205 U/L predicted nonawakening with a sensitivity of 49% and a specificity of 100%. Bilateral absence of the N1 peak predicted nonawakening with a sensitivity of 53% and a specificity of 100%. Using CKBB > or = 205 U/L, bilaterally absent SEP N1 peaks, or both predicted nonawakening with a sensitivity of 69% and a specificity of 100%. Using CKBB > or = 205 U/L, bilaterally absent N1 peaks, bilateral N3 > or = 176 msec or absent, or some combination predicted nonawakening with a sensitivity of 78% and a specificity of 100%. The combination of an absent N1 peak and elevated CKBB performs better than either alone in predicting nonawakening after cardiac arrest. Prolonged or absent N3 latency may increase sensitivity. These results should be interpreted with caution given the small number of patients and the possibility of a self-fulfilling prophecy.

“Non-hypercalcemic” analogs of 1α,25 dihydroxy vitamin D augment the induction of creatine kinase B by estrogen and selective estrogen receptor modulators (SERMS) in osteoblast-like cells and rat skeletal organs

We have demonstrated previously that daily treatments for 3 days with the so-called “non-hypercalcemic” analogs of 1α,25 dihydroxy vitamin D in ROS 17/2.8 osteoblast-like cells, stimulate the specific activity of creatine kinase BB (CK), and that such treatment with these analogs followed by a single treatment with gonadal steroids, upregulates responsiveness and sensitivity to estradiol 17β (E 2) for the induction of CK. This study was designed to determine if these same “non-hypercalcemic” vitamin D analogs could upregulate in vivo the response to E 2 and whether substitution of selective estrogen receptor modulators (SERMS) for E 2 would result in the same upregulation. We found that one week or 2 weeks pretreatment of prepubertal rats with vitamin D analogs led to increased induction of CK by E 2 and by the SERMS tamoxifen, tamoxifen methiodide and raloxifene, in epiphysis and diaphysis of the femur but not in the uterus. However, in contrast to their antiestrogenic activity in the uterus, there was no inhibition of E 2 action by the SERMS in skeletal tissues. The induction of mRNA for ckb in ROS 17/2.8 cells by E 2 or SERMS was demonstrated only after vitamin D pretreatment; there was no inhibition of E 2 induction by SERMS. Antagonists of vitamin D dependent calcium transport (transcaltachia) did not inhibit stimulation by vitamin D analogs. These results support the involvement of a nuclear mechanism in the upregulation of induction of CK by E 2, which may be due, in part, to the ability of vitamin D to increase estrogen receptor(s).

Amyloid beta-peptide (1-40)-mediated oxidative stress in cultured hippocampal neurons. Protein carbonyl formation, CK BB expression, and the level of Cu, Zn, and Mn SOD mRNA.

Mechanism of amyloid beta-peptide (A beta) toxicity in cultured neurons involves the development of oxidative stress in the affected cells. A significant increase in protein carbonyl formation was detected in cultured hippocampal neurons soon after the addition of preaggregated A beta(1-40), indicating oxidative damage of proteins. We report that neurons, subjected to A beta(1-40), respond to A beta oxidative impact by activation of antioxidant defense mechanisms and alternative ATP-regenerating pathway. The study demonstrates an increase of Mn SOD gene expression and the restoration of Cu, Zn SOD gene expression to a normal level after temporary suppression. Partial loss of creatine kinase (CK) BB activity, which is the key enzyme for functioning of the creatine/phosphocreatine shuttle, was compensated in neurons surviving the A beta oxidative attack by increased production of the enzyme. As soon as the oxidative attack triggered by the addition of preaggregated A beta (1-40) to rat hippocampal cell cultures has been extinguished, CK BB expression and SOD isoenzyme-specific mRNA levels in surviving neurons return to normal. We propose that the maintenance of a constant level of CK function by increased CK BB production together with the induction of antioxidant enzyme gene expression in A beta-treated hippocampal neurons accounts for at least part of their adaptation to A beta toxicity.

Nonhypercalcemic Analogs of Vitamin D Stimulate Creatine Kinase B Activity in Osteoblast-Like ROS 17/2.8 Cells and Up-regulate Their Responsiveness to Estrogens

We have reported that pretreatment with 1α, 25(OH) 2D 3(1,25) up-regulates responsiveness and sensitivity to 17β estradiol (E 2) in osteoblast-like cells, as measured by parallel stimulation of [ 3H]thymidine incorporation into DNA and the specific activity of creatine kinase BB (CK). Increased responsiveness was correlated with increased E 2 receptor concentration. In this study, we have extended these observations to new nonhypercalcemic analogs of 1,25. We compared the analogs hexafluoro vitamin D 3 (FL), and the side chain modified derivatives: EB 1089 (EB), CB 1093 (CB) and MC 1288 (MC) with 1,25 and 25 (OH)D 3(25 D 3). Treatment with 30 nM E 2 for 4 h stimulated CK activity in ROS 17/2-8 cells by 40%; there was no further increase after 3 daily additions of E 2. Treatment by 3 daily additions, at 1 nM, of all analogs except 25 D 3 caused a 2–3-fold increase in CK specific activity. This schedule of treatment also upregulated the response to 4 h exposure to 30 nM E 2 by 30–70% above the response to vitamin D analogs alone, and by up to 2 fold compared to E 2 without pretreatment. At 1 pM, the analogs doubled CK activity, and, except for 1,25, upregulated the response to E 2 to levels characteristic of each analog. Pretreatment with vitamin D analogs also increased the sensitivity to E 2 by lowering the dose for a comparable response to E 2 by one or two orders of magnitude. Stimulation of specific activity of CK by the analogs was paralleled by increases in the steady state level of mRNA for CKB, but not in its half life. Whereas pretreatment by vehicle followed by E 2 for 2 h was unable to increase CKB mRNA, pretreatment with the analogs made possible detection of mRNA responsiveness to E 2. These results add to the evidence for the interaction of estrogens and antiestrogens with vitamin D metabolites in regulation of bone growth in vitro. They also strengthen the potential for treatment of bone loss, as occurs in postmenopausal osteoporosis, by a combination of nonhypercalcemic vitamin D analogs and estrogens.

Effects of PTH-rP(107–111) and PTH-rP(7–34) on Adult Cardiomyocytes

We investigated whether parathyroid hormone-related peptide (PTH-rP), recently found expressed in the heart, exerts growth and contractile effects on adult cardiomyocytes from rat hearts. Synthetic PTH-rP peptides were used covering either a protein kinase C (PKC)-activating domain [PTH-rP(107–111)], or an adenylate cyclase activating domain [PTH-rP(1–34) and PTH-rP(7–34)]. PTH-rP(107–111) (1μm) increased creatine kinase BB activity (CK-BB), a CK isoform re-expressed during cardiac hypertrophy, within 24 h by 62±12%. This induction was abolished in the presence of the mitogen-activated-protein (MAP)-kinase-kinase inhibitor PD 98059. PTH-rP(107–111) activated p42-MAP-kinase within 15 min, increased protein synthesis (19± 4%), total protein mass (19±5%), cell volume (45±7%), and cross-sectional area (38±9%) of cardiomyocytes. Activation of p42-MAP-kinase and increase in protein synthesis were abolished in presence of bisindolylmaleimide, a PKC inhibitor. PTH- rP(107–111) did not directly influence contractile activity but reduced the contractile response to isoprenaline. In contrast, PTH-rP(1–34) and PTH-rP(7–34) induced spontaneous contractile activity in 3-day-old cultures. This induction was abolished in presence of Rp-cAMPS, a protein kinase A inhibitor, indicating an involvement of cAMP in this response. PTH-rP(1–34) also increased the cellular accumulation of cAMP. It is concluded that PTH-rP exert direct effects on adult cardiomyocytes by activating either PKC via a functional domain covered by amino acids 107–111 or by activation of cAMP-dependent protein kinase via a functional domain covered by amino acids 7–34. Since these parts of PTH-rP have either no homology [PTH-rP(107–111)] or only a limited structural similarity [PTH-rP(7–34)] to parathyroid hormone, these activities of PTH-rP have to be clearly distinguished from those described for parathyroid hormone.

Does a routine operation for intracranial aneurysm incur brain damage?

A biochemical marker of brain cell damage, the BB-isozyme of the intracellular enzyme Creatine Kinase (CK), was used to evaluate any possible injury to the brain, caused by an operation for a ruptured intracranial aneurysm (SAH). CSF-CK BB was assessed before and at intervals after operation in a series of 60 patients, aged 29-71 (mean 51 years) operated on for intracranial aneurysms, all but one after SAH. The m/f ratio was 18/42. 35 of the 60 patients were operated on acutely, i.e. within 72 hours after the SAH. CK BB was determined as CKB-activity after immunological inactivation of CKM. Normally there should be almost no detectable enzyme activity in the CSF. The pre-operative CK BB-activity was 0.01+ -0.01 mikrokatal in the patients in Hunt & Hess grade I who were operated on > 7 days after their SAH, and 0.05+ -0.04 in those operated on acutely, probably still reflecting the effects of the SAH on the brain. The mean per-operative CK BB increase was 0.11+ -0.17 for patients who had an uneventful postoperative course, compared to 0.39+ -0.49 for those showing some degree of immediate postoperative deterioration. This difference is significant at the 1% level. 52 of the 60 patients showed a rise of CK BB after operation. The mean increase for those patients operated upon in a good state and without any complication or postoperative deterioration was 0.02+ -0.03 mikrokatal, which could therefore be considered as a "normal" or acceptable elevation.(ABSTRACT TRUNCATED AT 250 WORDS)

Creatine kinase-BB: A marker of liver sinusoidal damage in ischemia-reperfusion

Cell damage within the sinusoidal lining of human liver grafts during transplantation is an early event that is critical in ischemia-reperfusion injury and probably plays a key role in primary liver dysfunction after transplantation. No simple biochemical marker for sinusoidal injury is currently available. Because creatine kinase activity has been described in heart endothelial cells, we hypothesized that release of this enzyme might serve as an index of sinusoidal injury. To test this hypothesis, we used several in vivo and in vitro experimental models. Occlusion of the rat hepatic pedicle in situ for 60 min (normothermic ischemia) induced a significant increase in serum creatine kinase levels relative to those in laparotomized controls (2,530 ± 530 vs. 389 ± 64 IU/L, mean ± SEM; p < 0.005). In the isolated perfused rat liver, 60-min ischemia induced early (≤ 3 min) creatine kinase and AST release (0.87 ± 0.14 vs. 0.08 ± 0.01 IU/min/gm liver, respectively). A similar phenomenon was observed after 24-hr or 48-hr hypothermic conservation in University of Wisconsin solution. Electrophoretic analysis and immunoinhibition studies showed that creatine kinase activity comprised creatine kinase-BB (≈ 50%) and mitochondrial creatine kinase. Trypan blue infusion showed a loss of viability in sinusoidal cells, whereas hepatocytes were relatively spared. Finally, murine sinusoidal cells were isolated, cultured and then lysed by a freeze-thaw cycle and sonication. Creatine kinase activity was found in endothelial cells (creatine kinase-BB), Kupffer cells (creatine kinase-BB) and Ito cells (creatine kinase-MM). Creatine kinase-BB was not found in hepatocytes, but mitochondrial creatine kinase was detected. These results indicate that liver ischemia-reperfusion injury induces a release of creatine kinase-BB from sinusoidal cells and of mitochondrial creatine kinase from hepatocytes, suggesting that creatine kinase-BB activity could provide a simple index of cell damage within the sinusoidal lining. (HEPATOLOGY 1993;17:423–428.)

Creatine Kinase-BB Activity in Malignant Tumors and in Sera from Patients with Malignant Diseases

Creatine kinase (CK EC 2.7.3.2) and CK-BB activity was analyzed in 41 malignant tumors of 6 different sites and different histological structures. The same analyses were done on 150 sera of patients with malignant diseases of various localizations. The rate of CK activity was determined kinetically, whereas tissue and serum CK-BB were separated chromatographically (Mercer). Insofar as malignant tumor tissues are concerned, the highest average rate of CK-BB activity was detected in tumors of the prostate (mean 1450 IU/g), and the lowest in tumors of the parotid gland (mean 5.2 IU/g). CK-BB was detected by the Mercer technique in 56 (37.3%) of 150 analyzed sera of patients with malignant diseases. The rate of CK activity in sera of patients with malignant diseases was 8 to 74 IU/I. In comparison with the site of the malignant process no significant CK serum activity differences were observed. T2-T3 tumors did not significantly influence the activity of either CK or CK-BB in the case of either tissues or sera (T1-T3). Enzyme activity was found to be much higher--both in tumoral tissue and in sera--with T4 tumors. The highest rate of CK-BB activity was found in sera of patients with malignant tumors of the stomach (mean 8.1 IU/I), and the lowest in malignant tumors of the rectum (mean 1.8 IU/l).

Improved outcome prediction based on CSF extrapolated creatine kinase BB isoenzyme activity and other risk factors in severe head injury.

The present study of 43 patients with severe head injury shows that outcome prediction can be markedly improved by combining an appropriate marker of the degree of initial brain damage and other risk factors. The patients were classified into three groups according to their actual outcome after 6 months: death (22 patients); persistent vegetative state or severe disability (eight patients); and moderate disability or good recovery (13 patients). By applying stepwise logistic discriminant analysis to the patients' data, five significant risk factors were selected: degree of neurological damage assessed by cerebrospinal fluid (CSF) extrapolated creatine kinase BB isoenzyme activity, Glasgow-Liège Coma Scale score, age, incidence of thoracic injury, and intracranial pressure (ICP). Extrapolated creatine kinase BB activity had the highest prognostic ability (67%). Uncontrollable elevated ICP proved to be systematically associated with death, whereas its absence was not necessarily indicative of a favorable outcome. The combination of the five variables yielded a total prognostic efficiency of 91%. The percentages of correctly predicted patients for the three outcome groups were, respectively, 100%, 50%, and 100%. Thus, half of the persistently vegetative and severely disabled patients were identified by the selected factors.

Creatine kinase BB activity in serum of patients with acute stroke: correlation with the severity of brain damage.

BB-CK activity was measured in 11 patients with stroke and in 10 controls. Blood samples were taken 36 hours after the clinical stroke onset in every patient. Sera were stored at -80 degrees and analyzed within two months. The creatine kinase isoenzymatic pattern was determined by ion-exchange column separation and gradient elution system. The mean BB-CK concentration in patients with stroke was significantly higher than in controls (p less than 0.01). In the group of "stroke" patients we found a correlation between severity of brain damage, as suggested by the clinical picture and CT scans, and serum values of BB-CK.

"Extrapolated" creatine kinase-BB isoenzyme activity in assessment of initial brain damage after severe head injury.

The severity of initial brain damage is an important risk factor in determining the prognosis of head trauma. It can be assessed by assigning neurological scores or by determining the cerebrospinal fluid (CSF) activity of the isoenzyme creatine kinase-BB (CK-BB). In 10 severely head-injured patients serial CSF samples were obtained during the first 24 hours after trauma, and exponential decay of CK-BB activity with an average half-life of 4.5 hours was demonstrated. This finding led the authors to propose an "extrapolated" CK-BB activity, which theoretically occurs immediately after injury and is calculated from a single CK-BB recording, as a new index for assessing the degree of initial brain damage. In 50 patients with severe head injury, the prognostic ability of "observed" and "extrapolated" CK-BB activity was compared with two clinical scoring systems that evaluate severity of head trauma (the Glasgow and the Glasgow-Liège Coma Scales). "Extrapolated" CK-BB activity proved to be the best prognostic factor. With a CK-BB cutoff point of 330 U/liter, a true-positive rate of 79% and a true-negative rate of 73% were obtained. These results suggest the usefulness of measuring CK-BB activity in CSF as soon as possible after hospital admission for head injury.

Regulation of creatine kinase activity in rat osteogenic sarcoma cell clones by parathyroid hormone, prostaglandin E2, and vitamin D metabolites.

We have previously shown that both parathyroid hormone (PTH) and prostaglandin E2 (PGE2) stimulate the activity of creatine kinase BB (CKBB) in rat bone cells in culture. Therefore, morphologically distinct rat osteogenic sarcoma cells in culture were tested for stimulation of CKBB activity by hormones that regulate skeletal tissues. PTH stimulated CKBB in the osteoblast-like clone ROS 17/2; 1 alpha,25(OH)2D3 inhibited this activity while PGE2, CT and 24R,25(OH)2D3 had no significant effect. PGE2 stimulated CKBB activity in the fibroblast-like clone ROS 24/1, which was unresponsive to PTH, CT and Vitamin D metabolites. 24R,25(OH)2D3 as well as PGE2 (but not PTH, CT or 1 alpha 25(OH)2D3) stimulated CKBB in clone ROS 25/1, suggesting that this fibroblast-like clone has some chondroblast-like character. Both PTH and PGE2 stimulated the brain type isoenzyme of CK (CKBB), although the osteogenic sarcoma cell clones contain a significant proportion of the muscle type of CK (CKMM). Thus, increased CKBB activity can serve as an additional characteristic marker for the action of steroid and polypeptide hormones and for prostaglandins.