Multiple cardiac contractile protein abnormalities in myopathic Syrian hamsters (BIO 53 : 58)

Hearts of genetically myopathic male hamsters (BIO 53 : 58) were studied at 1 month, 2 months, 3 months, 4 to 5 months and 7 months of age. The time course of alterations in the cardiac myofibrillar ATPase activity, the relationship of myofibrillar ATPase activity to free [Ca2+], myosin ATPase activity and the distribution of heavy chain myosin isoenzymes were evaluated. Mg2+-Ca2+ ATPase activity of cardiac myofibrils in myopathics was increased in 4 month and 7 month-old hamsters. Elevated Mg2+ ATPase activity was found as early as in 2-month-old hamster. However, there was no loss in the regulation of the myopathic myofibrillar assembly as measured by the PCa response (10(-7) M to 10(-4) M Ca2+). Scans of SDS electrophoresis slab gels of cardiac myofibrillar proteins from control (C) and myopathic animals (M) did not show any differences at any age group (1, 4 and 7 months). There was a significant decrease in myosin Ca2+ ATPase activity and actin activated Mg2+-ATPase activity at 4 to 5 months and 7 months of age in the myopathic hearts. At all ages in normal and myopathic animals cardiac myosin consisted of three isoenzymes, V1, V2 and V3. At all ages in controls and at 1 to 3 months in myopathics, V1 predominated and the isoenzyme distribution was V1 greater than V2 greater than V3. However, in myopathics at 4 to 5 months, the distribution was V1 = V3 greater than V2 and at 7 months was V3 greater than V2 greater than V1. Our experiments suggest alterations in different components of the contractile protein system that occur at different stages of myopathy.     

On the developmental properties and tissue interactions of hexokinase

The interactions of the isozymes of hexokinase with cellular structure have been studied in the major tissues of the mouse during development. Overall, these data provide a clear indication that interactions between hexokinase and cellular structure are appreciable in all major tissues and at all stages of development, and an analysis of the isozyme status of the enzyme in both soluble and bound compartments has been effected. Further evidence in support of the already well documented interaction of hexokinase I to subcellular material in adult brain and kidney tissues is provided and extended to show that such interactions are extensive in both these tissues throughout development. In addition, evidence is provided that considerable hexokinase II activity is present in mouse foetal tissues in both the soluble and bound fractions and this isozyme is also shown to be the predominant "bound" form of the enzyme in adult skeletal muscle. By contrast, hexokinase III and IV are shown to be largely located in the cytosolic fraction of liver. The metabolic implications of these enzyme-structural interactions during development are discussed, as is the possibility of a functional linkage between hexokinase, which is bound to the mitochondria, and other enzymic components of the glycolytic sequence.     

Variability in drug metabolism: importance of genetic constitution

In man wide variability exists in the rate of metabolism of drugs and among factors which contribute to this phenomenon genetic constitution is of major importance. The metabolism of a number of drugs is subject to polymorphism and the frequency distribution of particular pharmacokinetic parameters shows bimodality, with poor (PM) and extensive metabolizers (EM). Acetylation of a number of drugs is known to be polymorphic and the incidence of poor metabolizers varies markedly among different populations. Debrisoquine and sparteine are frequently applied model substrates for the characterization of a polymorphism in oxidative metabolism. Polymorphic drug oxidation may have important clinical implications, because when standard dosage regimens are applied plasma concentrations will reach far above the maximum acceptable in poor metabolizers and consequently side effects may arise. Regarding the multiplicity of the drug oxidizing enzyme system (cytochrome P-450) it could be of interest to combine model substrates in a cocktail to be able to characterize human subjects simultaneously for a number of independent polymorphisms.     

T-leukemia cell lines CCRF-CEM,HPB-ALL,JM and MOLT-4: Changes in isoenzyme profiles during induction of differentiation

Summary Biochemical analysis has been used to monitor the induction of differentiation in cultured human T-leukemia cell lines (CCRF-CEM, HPB-ALL, JM and MOLT-4) by the phorbolester 12-O-tetradecanoylphorbol 13-acetate (TPA). The isoenzymes of carboxylic esterase, acid phosphatase, hexosaminidase and lactate dehydrogenase were separated by isoelectric focusing on horizontal thin-layer polyacrylamide gels and stained by histo-cytochemical methods. TPA inhibited the proliferative activity in all four cell lines and led to aggregation of cells seen as floating clusters. TPA induced an increase in number and staining intensity of isoenzymes of all four enzymes in the cell lines studied. This corresponds to an induced isoenzymatic maturation as the progressive increase in number and staining intensity of the isoenzymes parallels the differentiation along the T-cell pathway. However, regardless of the initial stage of arrested differentiation, the cell lines could be induced only to differentiate to a certain more mature stage, but could not be triggered to differentiate terminally with regard to expression of isoenzyme patterns.     

Protein kinase C-β and -δ isoenzymes promote arachidonic acid production and proliferation of MonoMac-6 cells

In this study, we investigated the putative roles of certain protein kinase C (PKC) isoenzymes in the regulation of proliferation and arachidonic acid (AA) release in the human monocytoid MonoMac-6 cell line. Experiments employing specific PKC inhibitors and molecular biological methods (RNA-interference, recombinant overexpression) revealed that the two dominantly expressed isozymes, i.e., the "conventional" cPKCbeta and the "novel" nPKCdelta, promote AA production and cellular proliferation. In addition, using different phospholipase A(2) (PLA(2)) inhibitors, we were able to show that the calcium-independent iPLA(2) as well as diacylglycerol lipase (but not the cytosolic PLA(2)) function as "downstream" targets of cPKCbeta and nPKCdelta. In addition, we have also found that, among the other existing PKC isoforms, cPKCalpha plays a minor inhibitory role, whereas nPKCvarepsilon and aPKCzeta apparently do not regulate these cellular processes. In conclusion, in this paper we provide the first evidence that certain PKC isoforms play pivotal, specific, and (at least partly) antagonistic roles in the regulation of AA production and cellular proliferation of human monocytoid MonoMac-6 cells.     

First report of diffuse cutaneous leishmaniasis and Leishmania amazonensis infection in Rio de Janeiro State, Brazil

Diffuse cutaneous leishmaniasis (DCL) is characterised by multiple and progressive cutaneous lesions, resistance to chemotherapy and Leishmania-specific T-cell anergy. We report the first autochthonous DCL case and the first human infection with Leishmania amazonensis in Rio de Janeiro State, Brazil, where only L. braziliensis is considered to be the causative agent of cutaneous leishmaniasis. Leishmania amazonensis was identified by multilocus enzyme electrophoresis and PCR-RFLP. Our case was diagnosed as DCL according to clinical, parasitological, histopathological and immunological criteria. These observations indicate that L. amazonensis is increasing its geographical distribution in Brazil, accounting for unusual clinical presentations in new transmission areas.     

Characterization of Trypanosoma brucei s.l. subspecies by isoenzymes in domestic pigs from the Fontem sleeping sickness focus of Cameroon

Though it has been established that domestic animals (especially the pig) are potential reservoir hosts for Trypanosoma brucei gambiense in West Africa, there is little data to this effect concerning Central Africa. Instead, some previous authors report the absence of Trypanozoon type trypanosomes in domestic animals in Cameroon. Thirty-two domestic pigs were sampled by KIVI (kit for in vitro isolation) of trypanosomes in the northern region (Bechati) of the Fontem sleeping sickness focus of Cameroon. Twenty-one of these were found positive, from 15 of which 17 isolates were successfully obtained. Isoenzyme characterization revealed that isolates from 4 of the 15 pigs belonged to zymodemes associated with T. brucei gambiense group 1. The prevalence of this disease in the local human population is, however, very low. It is evident from this study that the domestic pig may be a potential reservoir host for T. brucei gambiense in the Fontem focus. There is, however, need for an extensive study on domestic animals in Cameroon and other neighbouring countries for a better comprehension of the epidemiology of sleeping sickness within the Central African region.     

35例高活性CK标本的CK-MB质量及活性检濒结果分析

目的探讨对高活性CK标本进行CK—MB质量及活性检测比较,为临床提供准确结果。方法将35例高活性cK标本分别在全自动化学发光仪及全自动生化分析仪上检测其CK—MB质量及活性。结果CK—MB活性的测定结果与实际值相比远远要高;CK—MB质量检测可有效减少检测干扰,有效保证可靠的检测结果。结论高活性CK标本严重影响其CK—MB活性检测,但对CK—MB质量检测影响不大。     

Significance of arginase determination in body fluids of patients with hepatocellular carcinoma and liver cirrhosis before and after surgical treatment

Objective

To assess the utility of arginase activity and expression in diagnosis of liver diseases.

Design and methods

Arginase activity, sensitivity and specificity were determined in serum of 140 patients including 50 with HCC, 60 with LC, 30 with choledocholithiasis (CDL) and 90 healthy controls. In HCC and LC arginase activity in serum was studied before and after tumor resection or liver transplantation. Arginase sensitivity in HCC was compared to that of alpha-fetoprotein (AFP) and aminotransferases (AST, ALT). In LC the activity was determined also in bile before and after transplantation. The expression of arginase isoenzymes in serum was studied by Western blotting.

Results

In HCC and LC the preoperative arginase activity was significantly higher compared to controls, and it decreased after surgery. The sensitivity of arginase in HCC was much higher than that of AFP, AST and ALT (96, 40, 20 and 18%, respectively). In HCC it was higher than in LC (93%) and CDL (33%). The specificity of arginase was above 80%. In bile of cirrhotic patients the highest activity was immediately after liver transplantation. It decreased with time but increased dramatically at the time of the graft rejection. Arginase AII was present in serum of HCC and LC but not the control cases.

Conclusions

The increase of arginase activity in serum accompanied by the presence of isoenzyme AII can be useful in HCC and LC diagnosis. The determination of arginase activity in bile may be helpful in monitoring liver graft recipients.