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Elevated serum CK-MB and CK-BB isoenzymes in a patient with small cell lung cancer: a case report.

An increase in serum creatine kinase-MB (CK-MB) isoenzyme is regarded as a specific indication of acute myocardial infarction. Recently, increased concentrations of various types of creatine kinase isoenzymes (mostly CK-BB and macro CK type 2 isoenzymes) have been found in the blood of some patients with malignancies. However, highly elevated serum CK-MB isoenzyme associated with malignancy has rarely been reported previously. Herein a case of extensive small cell lung cancer is presented in which the elevated serum levels of CK were detected throughout the three weeks of hospitalization. Fractionation of its isoenzymes by gel electrophoresis displayed elevated CK-MB and CK-BB isoenzymes. The proportion of serum CK-MB and CK-BB isoenzyme activities was persistently above normal (CK-MB 50-53%, normal < 5%). Clinical examination showed no evidence of myocardial infarction or injury, or tumor involvement of the heart. The tumor was the probable source of most circulating CK-MB isoenzyme. In clinical practice, markedly elevated levels of CK-MB, or increased levels of CK-MB in combination with CK-BB may point away from a myocardial origin and toward the existence of a malignancy.

Aged↗

Enzymatic method for quantitating lactate dehydrogenase isoenzyme composition in cardiac tissue: utilization of the method to characterize newborn and adult rat heart.

Lactate dehydrogenase (LDH) is an enzyme of special interest due to its key position in anaerobic metabolic pathways. Work on the structure and developmental expression of LDH led to formulation of the isoenzyme concept as well as elucidation of the biological significance of isoenzymes. Tissue isoenzyme compositional changes are mediated by changes in pattern of gene expression for two primary LDH gene loci in vertebrates. Developmental shifts in LDH gene expression prompted other experimental interventions to alter gene expression schedule and enzyme synthesis in order to pinpoint underlying genetic and molecular control mechanisms. LDH isoenzymes in tissues can be separated by electrophoresis and then quantitated by scanning densitometry; however, these methods require specialized instruments. Enzyme specific activity can be assessed spectrophotometrically, but enzymatic activity determination alone does not provide quantitation of LDH isoenzyme(s). The present work was conducted to establish a spectrophotometric enzyme assay procedure based on differential substrate inhibition to quantitate tissue LDH isoenzymes. The procedure was then used to assess developmentally related alterations in LDH isoenzymes in cardiac tissue of newborn and young adult rats. Results show that heart tissue in 5-day old newborn rats possesses approximately equal proportions of muscle type (anaerobic) and cardiac type (aerobic) LDH isoforms. As normal development transpires, heart tissue LDH isoenzyme proportion shifts with substantial decrease in anaerobic form which is accompanied by marked augmentation in aerobic form.

Aerobiosis↗

Creatine kinase isoenzymes in baboon tissues and organs.

We determined the activity of the isoenzymes of creatine kinase in homogenates of fresh tissues from six baboons. Twenty-six tissues were removed from each baboon and the isoenzymes in them were separated by chromatography on diethylaminoethyl-Sephadex A-50. Total activity was greatest in skeletal muscle (2600 U/g wet weight) followed by myocardium (800 U/g), brain (180 U/g), and the bladder and tissues of the gastrointestinal tract (about 100 U/g). Ten tissues had total activities of less than 20 U/g: trachea, lung, kidney, thyroid, gall bladder, liver, spleen, pancreas, common bile duct, and adrenal. Skeletal muscle contained only the MM isoenzyme; myocardium contained 81% MM isoenzyme, 18% MB isoenzyme, and 1% BB isoenzyme. Except for thyroid and pancreas, all other tissues contained mainly the BB isoenzyme. Tissues of the gastrointestinal tract contained significant (7 to 17%) MB isoenzyme activity.

Animals↗

[Lactate dehydrogenase isoenzymes in the serum and bronchial aspirate of newborn infants with respiratory difficulty of different etiologies].

OBJECTIVE: To study the usefulness of lactate dehydrogenase isoenzymes serum determination as tissue injury markers in newborns with respiratory distress. DESIGN AND METHODS: Ninety four neonates were studied and classified in two groups: 64 suffering various types of respiratory problems, and 30 healthy newborns of a similar birth weight and gestational age. LDH activity and its isoenzymes was determined in the serum of all the infants and in 23 samples of the bronchial aspirate of infants who required ventilation support. The isoenzymes were separated by electrophoresis on agarose gel and their activity was expressed as percentage of the total LDH. RESULTS: LDH1 and LDH2 isoenzymes were decreased, and LDH4 and LDH5 isoenzymes were significantly increased (p > 0.001) in infants serum with respiratory distress, compared with controls. We compared LDH isoenzymes values found in bronchial aspirate with their values found in serum of ventilate infants, and we found a significant levels of LDH2 and LDH3 were lower, and those of LDH5 were higher (p < 0.001) in bronchial aspirate than in serum and a positive correlation (r = 0.47, p < 0.01) between LDH5 values in both samples. CONCLUSIONS: The study shows significantly differences in the LDH isoenzyme profiles of neonates with respiratory distress compared with controls. The increase in serum of LDH4 and particularly of LDH5 isoenzymes could be an effective marker of tissue damage in lung disease in the newborn.

Acute Disease↗

Creatine kinase and its isoenzymes in the serum of women during pregnancy and the peripartum period.

In serum obtained from 28 women before, during, and after normal labor and delivery, creatine kinase activity was seen to be distinctly elevated immediately after labor and 24 h later, but had returned to normal six weeks later. In most cases the increase was due to the MM isoenzyme and was attributed to skeletal-muscle damage associated with labor. In 15 cases, the BB isoenzyme was observed, and in three patients the MB isoenzyme. Cord blood, which contains all three isoenzymes, may be the source of the MB and BB isoenzymes. Uterine muscle contains exclusively BB isoenzyme, and therefore uterine muscle damage is a likely source of the serum BB isoenzyme. The relationship among these isoenzymes in serum after delivery must be recognized, to avoid misdiagnosing myocardial infarction at this critical time.

Creatine Kinase↗

New sandwich ELISA for human urinary N-acetyl-beta-D-glucosaminidase isoenzyme B as a useful clinical test.

We have developed a new ELISA for quantifying N-acetyl-beta-D-glucosaminidase (NAG) isoenzyme B in human urine after raising monoclonal antibodies against the isoenzyme from human placenta. Though the obtained antibodies reacted not only to isoenzyme B but also to A, we could detect isoenzyme B selectively by a two-step sandwich ELISA with a pair of selected antibodies at low pH in the first reaction. The detected limit was 0.5 microgram/L for a sample volume of 25 microL. Within-run CVs ranged from 2.5% to 5.4% and between-run CVs ranged from 6.2% to 9.1%. Recoveries of NAG isoenzyme B added to each of three urine samples ranged from 91% to 114%. The dilution curves of urine samples showed good linearity. The cross-reactivity of NAG isoenzyme A was practically negligible (2-3%). The mean value for NAG isoenzyme B in spot urines from healthy adults was 2.9 micrograms/g creatinine. This ELISA method is rapid and precise enough for routine determination of NAG isoenzyme B in human urine.

Acetylglucosaminidase↗

Simultaneous detection of all four alkaline phosphatase isoenzymes in human germ cell tumors using reverse transcription-PCR.

We have developed a reverse transcription-PCR method that clearly distinguishes between the RNA transcripts of all four alkaline phosphatase (AP) genes. If compared to the methods used up to the present, the main advantages of the reverse transcription-PCR method presented are its specificity and high sensitivity. The germ cell AP and the placental AP, which are the two most closely related AP isoenzymes (98% homology), can clearly be distinguished without any interference by other AP isoenzymes. An enhanced expression of AP isoenzymes has been reported for various tumors. The examination of the pattern of AP isoenzyme expression in a specific tumor and the corresponding tissue of origin enables discrimination between eutopically and ectopically expressed isoenzymes and thus represents an important tool in the elucidation of AP isoenzymes as potential tumor markers. The pattern of AP expression in 15 germ cell tumors, 2 germinal epithelia adjacent to seminoma, 2 cell lines of germ cell tumor origin (Tera-1 and BeWo), and 5 normal testes was studied. In comparison to normal testes, in all seminomatous germ cell tumors eutopic expression of germ cell AP and ectopic expression of tissue-nonspecific AP were demonstrated. In both samples of pure embryonal carcinoma and in the embryonal carcinoma cell line, the transcription of all four mRNAs was shown. These results indicate that the expression of the isoenzymes depends on the degree of differentiation of a tumor and that a simultaneous up-regulation of all AP isoenzymes in all types of germ cell tumors does not exist.

Alkaline Phosphatase↗

Promotion of indoleacetic Acid oxidase isoenzymes in tobacco callus cultures by indoleacetic Acid.

Indoleacetic acid oxidase in tobacco callus cultures (Nicotiana tabacum L., cv. White Gold) was composed of at least two groups of isoenzymes, which were distinctly different in electrophoretic mobilities and in responses to growth substances. Indoleacetic acid had dual effects; at low concentrations it promoted the development of two fast-migrating indoleacetic acid oxidase isoenzymes, but at high concentrations it increased the level of other indoleacetic acid oxidase isoenzymes with low and moderate electrophoretic mobilities. However, indoleacetic acid was not unique in such effects; 2,4-dichlorophenoxyacetic acid and 2,4,5-trichlorophenoxyacetic acid were effective at concentrations lower than that of indoleacetic acid.A correlation was apparent between the relative levels of the two groups of indoleacetic acid oxidase isoenzymes and the rate and type of growth, as affected by auxins. The development of the fast-migrating anionic indoleacetic acid oxidase isoenzymes was accompanied by a type of growth characterized by rapid growth rate, high water content, and friable tissues. On the other hand, a further increase in the isoenzymes of slower migrating rate was associated with growth retardation.The indoleacetic acid-mediated increase of the fast-migrating indoleacetic acid oxidase isoenzymes was dependent on the level of kinetin, suggesting a multiple control by different types of growth substances. The inhibition of the formation of the fast-migrating isoenzymes by actinomycin D and cycloheximide suggests a requirement for both RNA and protein synthesis.

Journal Article↗

Malate dehydrogenase isoenzymes from cotton leaves: molecular weights.

Malate dehydrogenase isolated from leaves of the cotton plant (Gossypium hirsutum L.) appears in the form of several isoenzymes. Four of the isoenzymes found in cotton leaf extracts appear to be charge isomers with a molecular weight of approximately 60,000. A fifth malate dehydrogenase isoenzyme found in leaf extracts has a molecular weight of approximately 500,000. Under appropriate conditions it is possible to form this high molecular weight isoenzyme from at least one of the smaller isoenzymes. In addition, malate dehydrogenase isoenzymes of approximately 700,000 and 130,000 molecular weight have been observed under some conditions, although these isoenzymes do not appear in the crude cotton leaf preparations. The relationship of this heterogeneity with respect to size and to the discrepancies in the number and size of malate dehydrogenase isoenzymes reported from plant tissues may be significant.

Journal Article↗

Development of (1-->3,1-->4)-beta-d-Glucan Endohydrolase Isoenzymes in Isolated Scutella and Aleurone Layers of Barley (Hordeum vulgare).

An immunological assay has been used to investigate the synthesis of (1-->3,1-->4)-beta-glucanase (EC 3.2.1.73) isoenzymes from isolated barley aleurone layers and scutella. Enzyme release from both tissues is enhanced by 1 micromolar gibberellic acid and 10 millimolar Ca(2+), although increases induced by gibberellic acid are observed only in the presence of Ca(2+). Isoenzyme I is synthesized predominantly in the scutellum, while isoenzyme II is synthesized exclusively in the aleurone. A third, putative isoenzyme III has been detected in significant proportions in scutellar secretions and may also be secreted from aleurone layers. Both gibberellic acid and Ca(2+) appear to preferentially enhance isoenzyme II secretion from the aleurone and isoenzyme III secretion from scutella. The patterns of isoenzyme secretion are suggestive of tissue-specific differences in expression of the genes which code for (1-->3,1-->4)-beta-glucanase isoenzymes. Qualitatively similar results were obtained with barley cultivars harvested in Australia and North America.

Journal Article↗

Plant NAD(H)-Glutamate Dehydrogenase Consists of Two Subunit Polypeptides and Their Participation in the Seven Isoenzymes Occurs in an Ordered Ratio.

The structure and function of NAD(H)-glutamate dehydrogenase in plants was studied by using grapevine (Vitis vinifera L. cv Sultanina) callus grown under different nitrogen sources. The enzyme consists of two subunit-polypeptides, alpha and beta, with similar antigenic properties but with different molecular mass and charge. The two polypeptides have molecular masses of 43.0 and 42.5 kilodaltons, respectively. The holoenzyme is hexameric and is resolved into seven isoenzymes by native gel electrophoresis. Two-dimensional native/SDS-PAGE revealed that the 1 and 7 isoenzymes are homohexamers and the isoenzymes 2 through 6 are hybrids of the two polypeptides following an ordered ratio. The total quantity of alpha- and beta-polypeptides and the isoenzymic pattern was altered by the exogenous nitrogen source. The sample derived from callus grown on nitrate or glutamic acid contained a slightly greater amount of beta-polypeptide and of the more cathodal isoenzymes, whereas alpha-polypeptide and the more anodal isoenzymes predominated in callus grown in the presence of either ammonium or glutamine. The anabolic reaction was correlated with the alpha- and the catabolic reaction with the beta-polypeptide; this could suggest that each isoenzyme exhibits anabolic and catabolic function of different magnitude. The isoenzymic patterns did not obey the expected binomial distribution proportions.

Journal Article↗

Lactic dehydrogenase isoenzyme electrophoretic patterns in the diagnosis of pleural effusion.

The relative value of each lactic dehydrogenase isoenzyme (iso LDH) was measured by electrophoretic separation in the serum and the pleural fluid of 100 patients. In each case, the cause of the pleural effusion was known. Two types of LDH isoenzyme pattern were found in the serum: a normal type with a low value of LDH 5 and an abnormal type with a high value of LDH 5. This high LDH-5 level is due to an impaired liver function. In the pleural fluid, the electrophoretic patterns of five LDH isoenzymes were found by computerized processing. During congestive heart failure (28 cases) the electrophoretic pattern of the LDH isoenzymes was always similar in the serum and in the pleural fluid (transudative pleural effusion). During thoracic empyema, the relative values of the isoenzymes in the pleural fluid were regularly increasing from LDH 1 to LDH 5. In this situation, the evaluation of LDH 5 appeared to emanate from the increased granulocytes in the pleural fluid. In 22 inflammatory pleural effusions, the relative values of the five isoenzymes were equal. During malignant effusions (35 cases) a high level of LDH 5 was found in 21 patients. LDH 5 is known to be secreted by malignant tissue, and the authors confirmed it by finding a high level of LDH 5 in biopsy specimens of patients with mesothelioma or epidermoid lung cancer (7 cases). Conversely, the level of LDH 5 was low in biopsy specimens from normal lung tissue or benign inflammatory pleuritis (6 cases). Among the 14 patients with low levels of LDH 5 in the pleural fluid during malignant pleural effusion, the authors found the malignant lymphomas (three cases) and the small cell lung carcinoma (five cases). In these cases, the low level of LDH 5 was in agreement with the result of a low level of LDH 5 found in the biopsy of a metastatic liver localization of a small cell lung carcinoma. So the electrophoretic determination of LDH isoenzymes pattern in pleural fluid is a sensitive tool for the management of pleural effusion.

Carcinoma, Squamous Cell↗

Monocyte-specific esterase isoenzyme demonstrated by isoelectric focusing.

Using horizontal thin-layer isoelectric focusing in polyacrylamide gels, we separated the isoenzymes of carboxylic esterase (EC 3.1.1.1) of cell extracts prepared from human hematopoietic cells. Isoenzyme bands were visualized by staining with naphthol ester as substrate and coupling to an azo dye. Staining intensities of isoenzymes were quantified by densitometric scanning. On isoelectric focusing in a pH 2-11 gradient, distinct esterase isoenzyme profiles could be discerned and correlated to various types of normal hematopoietic cells and their leukemic counterparts. One unique isoenzyme, termed monoband, could be clearly identified on the basis of its isoelectric point (pI 6.0), its strong expression by normal and malignant monocytes and its complete and selective inhibition by sodium fluoride. This band was only found in monocytes of either normal or leukemic origin, but not in lymphoid or myeloid cells. The monocyte esterase could be inhibited by sodium fluoride whereas other isoenzyme bands were resistant to this inhibition. However, the specificity of this inhibitory reaction was relative, depending on the concentration of sodium fluoride. Compared with normal monocytes, leukemic monocytes often showed an overexpression of the mono-bands. Dilution experiments established the distinct prominence of the mono-band which could be detected among the other isoenzymes even when only 1% of the total cell population consisted of monocytes. Immature myeloid, but mono-band negative leukemic cells whose arrest of differentiation can be overcome by in vitro 12-O-tetradecanoylphorbol 13-acetate-promoted differentiation to more mature cells, could be induced to express the mono-band which paralleled their maturation to monocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Carboxylesterase↗

Monoclonal antibody specific to acid phosphatase isoenzyme 4.

Prostatic acid phosphatase isoenzyme 4 was purified by ion exchange column chromatography, followed by high pressure liquid chromatography. The highly purified enzyme was used to produce monoclonal antibody from immunized BALB/c mice. The antibody was specific to isoenzyme 4, with negligible affinity to isoenzyme 2. The specificity of the monoclonal antibody was evaluated by Western blot analysis and by inhibition of radioimmunoassay. Immunohistochemistry method using the antibody to isoenzyme 2 showed heavy staining on the cell surface in contrast to the even staining throughout the cytoplasm when monoclonal anti-isoenzyme 4 was used. These results reflect the secretory nature of isoenzyme 2 and the non-secretory nature of isoenzyme 4.

Acid Phosphatase↗

Tungstate can substitute for molybdate in sustaining growth of Methanobacterium thermoautotrophicum. Identification and characterization of a tungsten isoenzyme of formylmethanofuran dehydrogenase.

Methanobacterium thermoautotrophicum (strain Marburg) was found to grow on media supplemented with tungstate rather than with molybdate. The Archaeon then synthesized a tungsten iron-sulfur isoenzyme of formylmethanofuran dehydrogenase. The isoenzyme was purified to apparent homogeneity and shown to be composed of four different subunits of apparent molecular masses 65 kDa, 53 kDa, 31 kDa, and 15 kDa and to contain per mol 0.4 mol tungsten, < 0.05 mol molybdenum, 8 mol non-heme iron, 8 mol acid-labile sulfur and molybdopterin guanine dinucleotide. Its molecular and catalytic properties were significantly different from those of the molybdenum isoenzyme characterized previously. The two isoenzymes also differed in their metal specificity: the active molybdenum isoenzyme was only synthesized when molybdenum was available during growth whereas the active tungsten isoenzyme was also generated during growth of the cells on molybdate medium. Under the latter conditions the tungsten isoenzyme was synthesized containing molybdenum rather than tungsten.

Aldehyde Oxidoreductases↗

Foetal and maternal rat brain acetylcholinesterase: isoenzymes changes following insecticidal carbamate derivatives poisoning.

Pregnant rats (18th day) were orally given insecticidal carbamates with anticholinesterasic properties: 50 mg/kg carbaryl, 0.1 mg/kg aldicarb, 2.5 mg/kg carbofuran and 20 mg/kg pirimicarb. The acetylcholinesterase (AChE) isoenzymes from the brain of mothers and their foetuses were separated by electrophoresis on polyacrylamide ge. The four carbamate derivatives caused a significant lowering of the percentage of the least mobile isoenzyme (isoenzyme 1) in mother. Aldicarb, carbaryl and pirimicarb also caused a significant decrease in the percentage of the first foetal isoenzyme. The second foetal isoenzyme underwent a decrease after treatment with aldicarb or carbofuran. The distribution of the foetal isoenzymes was seen to be different to that of the dams in both control and treated animals. The difference in sensitivity of the cerebral AChE to the insecticides under investigation is thought to depend on differences of the fixation of carbamate derivatives on the foetal and maternal isoenzymes.

Acetylcholinesterase↗

Ultrathin-layer microelectrophoretic determination of lactate dehydrogenase isoenzymes in corneal and conjunctival epithelium of the cow.

In order to evaluate the impact of tissue oxygenation on the distribution pattern of lactate dehydrogenase isoenzymes, activities of the isoenzymes were measured in microdissected samples of bovine tissue. A highly sensitive ultrathin-layer electrophoretic technique was used to determine the distribution pattern of lactate dehydrogenase isoenzymes in basal, intermediate and superficial layers of the epithelium of central and peripheral cornea and in the epithelium of the bulbar conjunctiva. Measurements revealed almost homogeneous intraepithelial distribution patterns of lactate dehydrogenase isoenzymes in both tissues. In the cornea the lactate dehydrogenase isoenzymes 4 and 5, which are regarded to be specialized for anaerobic glucose metabolism, were found to predominate. In the well-oxygenated conjunctival epithelium most of the activity could be ascribed to the lactate dehydrogenase isoenzyme 3. In contrast to the isoenzymatic activities, total activity of lactate dehydrogenase was inhomogeneously distributed; maximum activities were found in the basal layer of corneal epithelium and in the intermediate layer of conjunctival epithelium. The results indicate that oxygen supply is relevant rather for the intraepithelial distribution of total enzyme activity than for the expression of lactate dehydrogenase isoenzymes.

Animals↗

Additional lactate dehydrogenase (LDH) isoenzymes in normal testis and spermatozoa of adult man.

Adult human testicular tissue contains up to six previously undescribed lactate dehydrogenase (LDH) isoenzymes in addition to the five LDH isoenzymes normally found and the sixth found in spermatogenic cells and spermatozoa, LDH-X. Additional LDH isoenzymes were also found in spermatozoa but not in seminal fluid or in serum. After electrophoresis one additional LDH isoenzyme of testicular tissue was localized between LDH-1 and LDH-2, two between LDH-2 and LDH-3, two between LDH-3 and LDH-4, and two between LDH-4 and LDH-5. These localizations indicate that the additional LDH isoenzymes are tetramers combining the A and B subunits of the five normal LDH isoenzymes and the C subunit of LDH-X. The additional LDH isoenzymes may be important in the metabolism of spermatogenic germ cells and spermatozoa.

Adult↗