Brain-type isoenzyme of creatine kinase was measured serially in 45 healthy and 22 severely asphyxiated term infants. The enzyme was measured in cord blood and in venous, capillary, or arterial blood at six to eight hours, 24 to 30 hours, and 72 to 80 hours after birth. In the healthy infants a brief rise of CK-BB occurred at six to eight hours; CK-BB activities were greater than 2.5 log-transformed standard deviations above the mean of the control values in ten of the asphyxiated infants and in none of the control infants. When normal CK-BB activity was used as a predictor of good neurologic outcome and elevated CK-BB as a predictor of subsequent neurologic abnormality, the outcome was predictable from the CK-BB activity in 17 of 22 cases (77%) and in 11 of the 12 survivors (92%). Eight of the 12 surviving infants had neonatal seizures and outcome was predictable from CK-BB activity in all cases. We conclude that serum CK-BB activity, especially when measured in cord blood and at six to 12 hours of life, correlates with neurologic outcome after severe asphyxia, and that measurement of CK-BB compares favorably with radionuclide and computerized tomographic scanning as a method of predicting neurologic outcome after asphyxia.

The concentration of creatine kinase BB isoenzyme (CK BB) was measured by radioimmunoassay in CSF from 306 patients with various neurologic disorders. Levels above 2.0 ng/mL were not found in patients without neurologic disease. Whereas mean CSF CK BB level was less than 2.0 ng/mL in groups of patients with systemic malignant neoplasms, latent syphilis, peripheral neuropathy, disk disease, polyradiculopathy, myelopathy, multiple sclerosis, neurodegenerative disease, encephalopathy, and hydrocephalus, it was elevated in groups of patients with convulsive disorder, CNS neoplasm, cerebrovascular disease, vasculitis, and meningoencephalitis.

The BB isozyme of creatine kinase is consistently present in cultured human fibroblasts and shows great variation in activity in long-term lymphoid lines. One mouse line tested, PG 19, had strong activity, but all other rodent lines tested did not express CK BB. Human and mouse CK BB can be expressed independently of each other in human--rodent somatic cell hybrids. There is some evidence that the structural locus for CK BB may be on chromosome 14, but the involvement of other chromosomes, expecially no.17, cannot be excluded. The MM and MB isozymes of creatine kinase were not seen in any human cultured cells.

An accurate and close follow-up of serum levels of thyroid hormones and various muscle markers (myoglobin, creatine kinase and its isoenzymes CK-MB and CK-BB, lactic dehydrogenase, alpha-hydroxybutyric dehydrogenase, etc.) was carried out in 10 hypothyroid patients on replacement therapy. The main muscle markers, creatine kinase and myoglobin, were elevated respectively in 9 and 6 subjects. In 1 male significant CK-MB traces were measured, while in 1 woman significant CK-BB amounts were assayed. Significant correlations between patients' thyroid hormones and the levels of the muscle parameters were found. The rate of normalization of thyroid hormones and muscle markers in relation to replacement therapy was also studied. Myoglobin and creatine kinase have proved to be the best indicators of the hypothyroid myopathy, since they are sensitive for the early detection of muscle involvement due to the metabolic disorder and are closely correlated to the metabolic conditions of patients.

This pilot study sought to determine if creatine kinase isoenzyme BB (CK-BB) levels might serve as a biological marker of injury severity in mild TBI (mTBI). This was a retrospective study of 64 soldiers with suspected mTBI seen in a combat support hospital in Mosul between March and August of 2007. Four of the 64 total samples were positive for CK-BB. One major trauma patient had a negative CK-BB. This yields a sensitivity of 11% and a specificity of 97%. Military Acute Concussion Evaluation (MACE) scores collected also did not appear to reflect extent of injury. Although the low sensitivity of CK-BB from this study does not support its use as an early marker of suspected mTBI, the result is not conclusive given the small sample and the possibility of isoenzyme degradation during transport. Although limited, the data collected on MACE scores warrant additional evaluation of whether this measure is clinically relevant.

Monoclonal antibodies have been raised against human heart- and brain-type creatine kinase (CK-MB and CK-BB). We used a low-affinity monoclonal antibody to develop a simple two-step immunoaffinity purification procedure for native CK-MB. Antibodies of higher affinity were used to construct specific two-site immunoradiometric assays for CK-MB and CK-BB. In the assay for CK-MB we used an 125I-labeled B subunit-specific antibody with an immobilized anti-M subunit antibody--adding either simultaneously, for a 1-h assay in which between 5 and 1000 ng of CK-MB per milliliter could be measured with an intra-assay CV of 4% to 20%, or sequentially, for a 2-h magnetic separation assay in which between 0.5 and 1000 ng/mL could be measured with an intra-assay CV of 14 to 19%. In both versions CK-BB up to 100 ng/mL and CK-MM up to 5000 ng/mL did not interfere. In the assay for CK-BB we used an 125I-labeled, reduced, and alkylated monoclonal antibody specific for the B subunit of CK-BB, and removed bound isoenzyme with a second immobilized monoclonal antibody specific for a different epitope on the B subunit. Total incubation time for this assay was 5 h. Intra-assay CV was 7.5 to 20% between 0.1 and 1000 ng/mL. CK-MB up to 1000 ng/mL and CK-MM up to 100 000 ng/mL did not interfere. Inter-assay CVs in all three assays varied from 9 to 21%.

Fusion of splenocytes from A/J mice immunized by creatine kinase (EC 2.7.3.2)-MB isoenzyme (CK-MB) with SP2/0-Ag14 myeloma cell line generated hybridomas producing monoclonal antibodies specific to CK-MB. One of these monoclonal antibodies ("Conan-MB") was used to develop a direct assay for CK-MB activity. In the assay, serum is incubated for 30 min at room temperature with "Conan-MB" immobilized on latex beads. The beads are then washed, and CK-MB activity bound to the antibody is measured after incubation with CK enzyme reagent for 30 min at 37 degrees C. Results with the assay correlated well (r = 0.997) with those for CK-MB concentration as measured by a two-site immunoassay. Neither CK-MM, CK-BB, mitochondrial CK, nor a hemolysate of erythrocytes interfered. Use of this unique monoclonal antibody allows rapid, precise, and direct determination of CK-MB activity.

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.

Isoenzymes of creatine kinase (ATP:creatine phosphotransferase; EC 2.7.3.2; CK) were measured by electrophoresis in serum from cord blood and skin-puncture blood taken from 45 healthy full-term infants during the first three postnatal days. Mean total CK activities (in U/L at 30 degrees C) were 185 in cord samples, 536 in samples taken between 5--8 h postnatally, 494 between 24--33 h, and 288 in the 72-100 h samples. Values for all three isoenzymes increased to a peak over this period, with the highest values generally being found in the samples taken 5--33 h after birth; the subsequent decline was most rapid for CK-BB. Serum CK isoenzymes in cord samples and those taken at 72--100 h in the 11 babies delivered by cesarian section did not differ significantly from those of babies delivered vaginally. However the postnatal increases in total CK, CK-MM, and CK-MB (but not in CK-BB) were significantly greater in those patients born by vaginal delivery. The reasons for the increases in CK isoenzymes after birth are not clear, but our results and reported studies on the ontogeny of CK suggest that CK-MB cannot be regarded as a "cardiac-specific" isoenzyme in the neonatal period.

We have demonstrated previously that daily treatments for 3 days with the so-called "non-hypercalcemic" analogs of 1alpha,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 17beta (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).

We have previously reported that estradiol (E2) 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 E2 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 F2-2 (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 E2 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 E2 concentration, but prevented the suppression of DNA synthesis exerted by high concentrations of E2 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 D3, and JKF decreased the expression of ERbeta proteins in SMC and increased the ERalpha 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.

OBJECTIVE

The development of secondary brain injury after trauma is known to involve in many cellular mediators. The aim of the study was to evaluate and compare the effects of the use of both methylprednisolone and montelukast on serum and tissue concentrations of NO, malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity, and tissue glutathione peroxidase (GSH-Px) activity in rats with spinal cord injury (SCI).

METHODS

SCI was induced in Wistar albino rats by dropping a 10 g rod from a 5.0 cm height at T9-10. The 28 rats were randomly divided into four equal groups: montelukast, methylprednisolone, non-treatment and sham groups. Rats were neurologically tested at 24 hours after trauma and spinal cord tissue levels of MDA, SOD, GSH-PX, CAT levels and blood CK, CK-BB, LDH levels were measured. In addition, histopathological changes were also examined.

RESULTS

There was a significant improvement in Tarlov scores in methylprednisolone and montelukast administered group compared to the trauma group (p = 0.001). When compared to trauma group, methylprednisolone and montelukast groups had significant differences in MDA (p < 0.05), SOD (p < 0.001), CK-BB (p < 0.001) and LDH (p < 0.05) levels. Histopathologically, no significant changes were observed.

CONCLUSIONS

The present study shows effects of montelukast with biochemical and histopathological parameters and compares its effects with those of methylprednisolone for the first time. Our research has shown that montelukast and methylprednisolone have a neuroprotective effect on spinal cord injury.

We examined the biologic properties of a small-cell-lung-carcinoma (SCLC) cell line (designated MN-1112) established from a patient with SCLC who showed paraneoplastic retinopathy syndrome. Morphologic and immunocytochemical analyses showed that MN-1112 cells possess features of the classic type of SCLC. MN-1112 cells grew in suspension forming relatively large clumps of cells with a doubling time of 72 hr. By light-microscope examination, the cells were relatively small and had scanty cytoplasm. The cells produced NSE, ACTH and CK (BB isozyme); they also expressed recoverin, a novel photoreceptor protein, detected by Northern-blot and Western-immunoblot analysis using human-recoverin-specific DNA probe and anti-bovine-recoverin polyclonal antibody. This report shows that human recoverin is expressed in cultured SCLC cells. Our results support the hypothesis that, in cancer-associated retinopathy (CAR) patients, auto-immune antibody targeting for ectopic recoverin in SCLC is initially produced and cross-reacts with the retinal protein, resulting in the retinal degeneration that occurs in CAR patients.

Creatine kinase (CK), isocitrate (ICDH) and glucose-6-phosphate dehydrogenase (G-6-PD) activities and cytosol oestrogen (REc) and progestin (RPc) receptors have been measured in twenty post-menopausal subjects with endometrial carcinoma. Diagnostic curettage and hysterectomy specimens were obtained from each patient with intervening medroxyprogesterone acetate therapy (MPA). In RPc rich tumours the activity of creatine kinase was significantly increased following MPA and was attributed to an increase in the BB-isoform. There was a similar though less significant increase in ICDH activity. Neither CK nor ICDH were significantly increased following MPA therapy in progesterone receptor poor specimens. There was a small but significant increase in G-6-PD activity following MPA regardless of RPc content. REc was decreased in some but not all RPc rich tumours following MPA and the trend was significant. The highly significant increase in CK activity confirms the apparent response of this enzyme to progesterone in normal human endometrium during the menstrual cycle.

OBJECTIVE

To develop a highly sensitive immunofluorometric procedure for creatine kinase BB isoenzyme and use it to measure CK-BB in tumor cytosolic extracts and serum of cancer patients and healthy volunteers.

METHODS

For assay development, we used two monoclonal antibodies in combination with time-resolved fluorometry and the biotin-avidin system. We measured CK-BB in breast tumor cytosols and studied its association with steroid hormone receptors. We also measured CK-BB in the serum of healthy subjects and patients with prostate cancer. We have examined the molecular weight of CK-BB in serum using high performance liquid chromatography.

RESULTS

The evaluation of the method revealed good precision and accuracy. Study of 336 breast tumor cytosols and 9 normal breast cytosols has shown that CK-BB is overexpressed by 95% of breast tumors and that CK-BB is present in its 80 kDa form. A close association between CK-BB and estrogen but not progesterone receptors was found, suggesting that CK-BB overexpression is another marker of estrogen sensitivity of these tumors. Previous studies, using CK-BB radioimmunoassay could not detect CK-BB in the serum of about 50% of healthy subjects. We have assessed CK-BB levels in 80 male volunteers, detected CK-BB in all sera and provided a detailed distribution of values. We further demonstrated that 30% of prostate cancer patients in remission (PSA < 0.4 microgram/L) post radical prostatectomy and 50% of patients with active prostate cancer (PSA>> 20 microgram/L) have elevated serum CK-BB levels. The patients with highly elevated CK-BB also had highly elevated serum PSA. We have demonstrated that some patients who have elevated serum CK-BB also have macromolecular CK complexes in their serum with molecular weights of 700 and 350 kDa as well as the 80 kDa CK-BB isoenzyme. Only the latter was recognized by the assay developed.

CONCLUSIONS

CK-BB is a marker of estrogen sensitivity in breast cancer; Patients with prostate cancer have elevated CK-BB in their serum; The new highly specific and sensitive assay may be further used to study the role of CK-BB in various malignancies.

Postmortem biochemical indices may provide a useful adjunct to morphological studies in the identification of antemortem brain insult. We studied 34 routine medico-legal cases categorising them into one of four diagnostic groups. There were 11 cases of head trauma, 7 of 'hypoxia' (3 hangings and 4 carbon monoxide or drug poisonings), 7 sudden cardiac deaths and 9 miscellaneous cases. Survival time and postmortem interval was known for each case. The degree of cranio-cerebral trauma was graded. Cerebro-spinal fluid (CSF) and vitreous humour were analysed for calcium, glucose, total proteins, aldolase, aspartate transaminase (AST), alanine transaminase (ALT), gamma glutamyltransferase (GGT), lactate dehydrogenase (LDH), creatine kinase (CK) and creatine kinase BB isoenzyme (CK-BB). CK-BB was also measured in superior vena cava serum. In CSF there was a significant correlation between the severity of cranio-cerebral trauma and levels of aldolase, CK-BB, AST, ALT and total proteins. CSF CK-BB, median units/l (range), for the groupings of head trauma, hypoxia, sudden cardiac death and miscellaneous were respectively 823 (2-3431); 96 (2-187); 4 (2-25); 5 (1-69). Corresponding serum CK-BB levels were 240 (28-322); 390 (26-411); 180 (20-482); 79 (18-530).

We present analytical and clinical studies of three commercial immunoassays for the determination of the creatine kinase MB isoenzyme (CK-MB). All methods are compared with the presently-accepted reference method, agarose gel electrophoresis. We find that each of the immunoassay methods offers diagnostic sensitivity and specificity similar to those of electrophoresis, while none experiences detectable interference from high concentrations of the creatine kinase BB isoenzyme (CK-BB) or the so-called "atypical" bands. The presence of atypical bands can cause electrophoresis to yield inaccurate quantification or even misidentification of CK-MB. Because the immunoassay approaches are more rapid than electrophoresis and require less technical expertise, it is possible to evaluate patients in a more timely and cost-effective manner. Further, since immunoassay results can be available in approximately 1/2 hour, rapid real-time decisions can be made regarding the desirability of initiating or continuing such valuable but dangerous procedures as thrombolytic therapy.

The early detection of acute myocardial infarction (AMI) upon the onset of chest pain symptoms is crucial for patient survival. However, this detection is challenging, particularly without a persistent elevation of ST-segment reflected in an electrocardiogram or in blood tests. A majority of the available point-of-care testing devices allow accurate and rapid diagnosis of AMI. However, AMI diagnosis is reliable only at intermediate and later stages, with myocardial injury (> 6 h) and MI, based on the expression of specific cardiac biomarkers including troponin I or T (cTnI or cTnT), creatine kinase-MB (CK-MB), and myoglobin. Diagnosis at the early myocardial ischemia stage is not possible. To overcome this limitation, a sensitive and rapid microfluidic paper-based device (µPAD) was developed for the simultaneous detection of multiple cardiac biomarkers for the early and late diagnosis of AMI. The glycogen phosphorylase isoenzyme BB (GPBB) was detected during early (within first 4 h) ischemic myocardial injury. On the same µPAD platform, detection of prolonged elevation of levels of cTnT and CK-MB, which are only produced 6 h after the onset of chest pain in human serum, was possible. Sandwich immunoassay performed on the µPAD achieved reproducibility (RSD approximately 10% and intra-and inter-day precision (CV 10-20%, 99th percentile), as well as consistently stable test results for 28 days, with strong correlation (r²= 0.962), using the standard Siemens Centaur XPT Immunoassay system. The present findings indicate the potential of the µPAD platform as a point-of-care device for the early diagnosis and prognosis of AMI.

Creatine kinase (CK; EC 2.7.3.2) isoenzymes and their substrates have an important function in cellular energy generation and utilization. The brain isoform (CK-BB) has been implicated in cellular transformation processes involving the oncogenic products of the Ela virus and the p53 tumor suppressor gene. Cyclocreatine, an analogue of creatine, has been previously shown to inhibit the growth of a broad spectrum of cancer cells derived from solid tumors. Results reported herein indicate an increased level of creatine kinase activity in human prostate carcinoma cell lines and inhibitory effects of cyclocreatine alone and in combination with adriamycin on the growth of these cells in vitro and in vivo, in immune-deprived mice. Our results suggest the possible use of cyclocreatine in the treatment of prostatic carcinoma.

Twenty-two patients with subarachnoid haemorrhage were investigated for changes in myoglobin, total CK, CK-MB and CK-BB in serum and for the incidence of ECG abnormalities. Serial ECG's showed abnormalities in 20 patients; 15 of these had T wave changes, 15 Q-Tc prolongation, ten had S-T depression and nine U waves and in seven cases arrhythmias were found. The purpose of the study was to find out whether a relationship could be established between the ECG abnormalities and changes in serum myoglobin and enzymes. However, in no patient could myoglobin or enzyme patterns consistent with acute myocardial or cerebral damage be observed and therefore the ECG abnormalities do not seem to be related to detectable myocardial damage.

The aim of this study was to assess the effects of minocycline on cerebral ischemia-reperfusion (I/R) injury in rats. The study was carried out on 24 male Wistar albino rats, weighing 200-250 g, which were divided into three groups: (i) control (n = 8), (ii) I/R (n = 8) and (iii) I/R + minocycline (n = 8). Minocycline was administrated at a dose of 90 mg/kg p.o. to the I/R group 48, 24 and 1 h before ischemia. Following bilateral exposure of the common carotid arteries by anterior cervical dissection and separation of the vagus nerve, I/R injury was performed by occlusion. Following reperfusion, malondialdehyde (MDA), superoxide dismutase, glutathione peroxidase and catalase levels in the blood and brain tissue, and creatine kinase (CK), CK-BB, lactate dehydrogenase (LDH), neuron-specific enolase (NSE) and protein S100β levels in the blood were measured and the histopathological changes were monitored. Regarding histopathological evaluation, symptoms of degeneration were significantly improved in the I/R + minocycline group compared to the I/R-only group. Statistical analysis of the biochemical parameters revealed significant differences in MDA (p < 0.001), nitric oxide (p < 0.05), CK (p < 0.05) and CK-MB (p < 0.05) levels between the I/R + minocycline group and the I/R group. According to the literature, the effect of minocycline is firstly assessed by LDH, CK-MB, NSE and S-100β analysis in addition to antioxidant status and histopathological analysis.

Medulloblastoma cell line TE671 was characterized by morphologic, cytochemical, neurochemical, and growth criteria. In contrast to the uniform, in vivo histopathologic appearance of the tumor, TE671 in vitro exhibits six morphologic subtypes (Types I-VI) in varying percentages over 14 days in culture. TE671 grows as a monolayer by the merging of separate foci. Cells were positive for Periodic acid Schiff (PAS) and reticulum, and negative for the glial marker, glial fibrillary acid protein (GFAP). Receptors for human C3b (EAC) were present on 19% of the cells. Neural associated isoenzymes, neuron specific enolase (NSE) and creatine kinase (CK-BB) were demonstrated in TE671. Progeny of a single clonogenic cell manifested the morphologic heterogeneity of cell types (I-VI). The absence of markers specific for glial cells suggests that TE671 is an early (less differentiated) precursor. TE671, the only continuous human medulloblastoma cell line, provides an experimental model with which to compare and identify the subpopulation of neoplastic cells in medulloblastoma ex vivo.

Creatine kinase (CK) plays a crucial role in cardiac energy transduction. During chronic cardiac stress conditions leading to hypertrophy and/or heart failure, the profile of CK isoenzyme activities changes towards a fetal pattern with increases of BB- and MB-CK and decreases of MM-CK and mito-CK. Changes of myocardial CK gene expression are only indirectly reflected by measurements of CK activities. The purpose of this work was, therefore, to determine myocardial expression of B-, M- and sarcomeric mito-CK genes in an animal model of heart failure where hemodynamic alterations and CK system changes are well defined, that is, in the rat heart post-myocardial infarction. Intact residual left ventricular myocardium was harvested 2 months following infarction (MI; n = 7) or sham operation (sham; n = 6) after in vivo left-ventricular end-diastolic pressure (LVEDP) was recorded. Total CK activity was measured spectrophotometrically, CK isoenzyme distribution with agarose gel electrophoresis. Steady state mRNA levels coding for B-, M- and mito-CK genes were measured with quantitative PCR and were normalized for GAPDH expression. Total CK activity tended to be reduced in MI (5.51 +/- 0.62 IU/mg protein) compared to sham (6.77 +/- 0.24; P = 0.55). CK isoenzyme distribution showed an increase of fetal BB- + MB-CK (MI 22.0 +/- 3.1%, sham 15.1 +/- 1.0%; P < 0.05), no change of MM-CK and a decrease of mito-CK (27.0 +/- 1.5% sham, 20.8 +/- 2.0% MI: P < 0.05). Relative B-CK mRNA levels increased (sham 0.46 +/- 0.06, MI 1.03 +/- 0.09; P < 0.05) and M-CK mRNA levels decreased (sham 1.06 +/- 0.08. MI 0.66 +/- 0.09; P < 0.05) significantly post-MI. The increase of B-CK mRNA (r = 0.72; P = 0.009) and the decrease of M-CK mRNA (r = 0.76; P = 0.003) correlated significantly with in vivo LVEDP. Mito-CK mRNA levels remained unchanged after MI (sham 0.94 +/- 0.16, MI 0.98 +/- 0.09). Intact residual left-ventricular myocardium post-MI is characterized by increased B-CK-mRNA and reduced M-CK-mRNA expression.

Carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and the BB isoenzyme of creatine kinase (CK-BB) were evaluated before therapy in the sera of 195 patients with histologically confirmed small cell lung cancer (SCLC) in a prospective multicenter trial. Forty-four percent (84 of 193) of all patients had CEA levels higher than 5 ng/ml, 66% (111 of 168) had NSE levels higher than 12.5 ng/ml, and 32% (40 of 123) had CK-BB levels higher than 10 ng/ml. Clear pathologic levels were less frequently observed. Significantly higher pretreatment titers for CEA, NSE, and CK-BB were found in patients with bone marrow and/or liver metastases. The most elevated marker levels were observed in the group of nonresponding patients with bone marrow and/or liver metastases. Only a slight correlation between the pretreatment CEA level and survival time could be observed. Patients with pathologic NSE (greater than or equal to 30 ng/ml) levels and, in particular, those with pathologic CK-BB (greater than or equal to 25 ng/ml) levels had a significantly shorter median survival than those with normal or elevated levels. In addition, a positive linear correlation between pretreatment NSE and CK-BB (n = 116, r = 0.54) levels was found, but CEA levels did not correlate with other marker levels. From these data it is concluded that pretreatment CEA, NSE, and CK-BB levels are helpful in the clinical management of a subset of patients with SCLC, i.e., those with bone marrow and/or liver metastases.