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Adenosine deaminase activity was shown to decrease in each skeletal muscle type (the slow-twitch oxydative, fast-twitch oxydative - glycolytic and fast-twitch glycolytic) at the beginning of exercise of moderate intensity and to return to the control when exercise was continued till exhaustion. 5 min occlusion of the femoral artery had no effect on the enzyme activity in either muscle. The reduction of the enzyme activity at the onset of exercise could result in reduction of adenosine breakdown and thus contribute to vasodilation at this stage of increased contractile activity of the muscles.
Adenosine deaminase (ADA) activity was estimated in mucosal specimens obtained endoscopically from the duodenal bulb. Three groups of subjects were studied: 1. 9 patients with achlorhydria, 2. 12 subjects with normal gastric acid secretion, 3. 5 patients with hypersecretion. Enzyme activity was measured by determination of ammonia liberated from the substrate according to the Chaney and Marbach method. In patients with hypersecretion the ADA activity was lower than in those with achlorhydria (p < 0.001) and normal acid secretion (p < 0.02). A significant negative correlation between ADA activity in the duodenal bulb mucosa and basal and maximal gastric acid outputs was found. The present study seems to indicate a possible relationship between gastric acid secretion and duodenal ADA activity.
Seven healthy dogs were administered with methylprednisolone orally (2 mg/kg body weight/d) at the beginning of the study (between 0-10 d), then the dosing was decreased at 10 d intervals throughout 10 to 60 d in an alternate-day therapy programme. Drug administration to the dogs was stopped on the 60 d. Total leukocyte count; polymorphonuclear leukocytes, and neutrophil ratios increased and eosinophil, mononuclear leukocytes, lymphocyte, and monocyte ratios were decrease when these values were compared to changes between days. Furthermore, in lymphocyte subsets, B lymphocytes and CD 8 T lymphocytes ratios were found to decrease. In addition, CD 3 T lymphocyte ratios decreased on the 61 d and CD 4 T lymphocyte ratios increased on the 21, 41, 51 and 61 d. IgM and IgG levels decreased between the days. Serum total ADA and ADA 1 activities decreased during the application, but ADA 2 activity did not change significantly. As a result, after immunosuppressive dose of methylprednisolone application, reduction in the ADA and ADA 1 activity can be used as determinant of the suppression of immune system components.
Kinetic and thermodynamic studies were made on the effect of caffeine on the activ­ity of adenosine deaminase in 50 mM sodium phosphate buffer, pH 7.5, using UV spectrophotometry and isothermal titration calorimetry (ITC). An uncompetitive inhi­bition was observed for caffeine. A graphical fitting method was used for determina­tion of binding constant and enthalpy of inhibitor binding by using isothermal titra­tion microcalorimetry data. The dissociation-binding constant is equal to 350 uM by the microcalorimetry method, which agrees well with the value of 342 uM for the inhi­bition constant that was obtained from the spectroscopy method. Positive dependence of caffeine binding on temperature indicates a hydrophobic interaction.
Adenosine 5'-triphosphate (ATP) and adenosine are the crucial endogenous signaling molecules in immunity and inflammation. In this study we identified the source of extracellular adenosine in human B lymphoblasts, and evaluate the ATP release and metabolism. We observed that the B cells continuously released substantial quantities of ATP (35 pmol/106 cells) when subjected to slow motion in the incubation medium. The adenosine level in the B cell incubation medium was very low, and increased (5-fold) upon inhibition of adenosine deaminase activity with 10 µM of 2-deoxycoformycin (DCF). Inclusion of an inhibitor of equilibrate nucleoside transport (nitrobenzylthioinosine) in the incubation medium in the presence of DCF resulted in the elevation of adenosine level by 9-fold. Inhibition of ecto-ATPase activity with 100 µM of ARL67156 was associated with a 2-fold increase of the extracellular ATP level and a 3-fold decrease of adenosine concentration in the cell culture media. Inclusion of ,ß-methyleneadenosine 5'-diphosphate, a selective inhibitor of ecto-5'-nucleotidase in the incubation medium resulted in a significant decrease (7-fold) the adenosine concentration. In conclusion, our results indicate that ATP released from the B cell is the primary source of peripheral adenosine, and that the activities of ecto enzymes and the efficiency of Ado uptake through the nucleoside transporters determine the Ado level on the B cell surface.
The aim of the study was to investigate the importance of ADA serum and its isoenzyme activity in diagnosing white muscle disease in lambs. The animal material used in the study was forty seven Akkaraman lambs - twenty seven of which had clinical signs of white muscle disease (white muscle disease group) and twenty healthy lambs (control group). Blood samples were taken and Se levels, CK, LDH, AST, ADA and ADA izoenzyme activity were determined. Following this, the animals in the white muscle disease group received 1 mg sodium selenit + 60 mg vitamin E intramuscularly as a single dose. Fourteen days following sodyum selenit + vitamin E application blood samples were again taken and the same analysis repeated. Apart from ADA2 activity, other enzyme activities were significantly high (p<0.001), and serum Se concentrations were significantly low (p<0.001) in animals of the white muscle disease group compared to the control group. When the parameters obtained from the white muscle disease group were compared before and after treatment it was indicated that all enzyme parameters decreased significantly (p<0.001) after treatment. On the other hand, serum Se concentration increased (p<0.01) after treatment in the diseased group. The results confirm that determining ADA serum and its isoenzymes, together with CK, LDH and AST values seems to be useful in diagnosing white muscle disease.
In diabetes several aspects of immunity are altered, including the immunomodulatory action of adenosine. Our study was undertaken to investigate the effect of different glucose and insulin concentrations on activities of adenosine metabolizing enzymes in human B lymphocytes line SKW 6.4. The activity of adenosine deaminase in the cytosolic fraction was very low and was not affected by different glucose concentration, but in the membrane fraction of cells cultured with 25 mM glucose it was decreased by about 35% comparing to the activity in cells maintained in 5 mM glucose, irrespective of insulin concentration. The activities of 5'-nucleotidase (5'-NT) and ecto-5'-NT in SKW 6.4 cells depended on insulin concentration, but not on glucose. Cells cultured with 10-8 M insulin displayed an about 60% lower activity of cytosolic 5'-NT comparing to cells maintained at 10-11 M insulin. The activity of ecto-5'-NT was decreased by about 70% in cells cultured with 10-8 M insulin comparing to cells grown in 10-11 M insulin. Neither insulin nor glucose had an effect on adenosine kinase (AK) activity in SKW 6.4 cells or in human B cells isolated from peripheral blood. The extracellular level of adenosine and inosine during accelerated catabolism of cellular ATP depended on glucose, but not on insulin concentration. Concluding, our study demonstrates that glucose and insulin differentially affect the activities of adenosine metabolizing enzymes in human B lymphocytes, but changes in those activities do not correlate with the adenosine level in cell media during accelerated ATP catabolism, implying that nucleoside transport is the primary factor determining the extracellular level of adenosine.
The activity of adenosine deaminases (EC.3.5.4.4) in granulocytes and lymphocytes of patients with stable angina pectoris was lower by about 27% and 24%, respectively as compared with control group, whereas these values in erythrocytes and blood plasma were at the normal level.
The conversion of right-handed dsDNA and dsRNA to the left-handed Z-con - formation in volves a re or ga ni za tion of the nu cleo tides rel a tive to each other. This con- ver sion can be fa cil i tated by the tight bind ing of a Z-conformation-specific pro tein do­main from the ed it ing en zyme dsRNA adenosine deaminase. This may in flu ence the mod i fi ca tion of both pre-mRNAs as well as some replicating RNA viruses.
Adenosine deaminase (ADA) was partially purified 486- and 994- -fold from rat liver mitochondria and cytosol, respectively. Relative molecular mass of the enzymes from both fractions was 34 (KM). Km for adenosine and 2'-deoxy-adenosine were 3.08 x 10~5 M and 3.03 x 10 ~5 M for mitochondrial ADA and 3.12 x 10"5 M and 2.87 x 10~5 M for cytosolic ADA. The enzyme from both subcellular fractions had the maximum activity at pH 7.5 - 8.0, and pi 5.2 and 4.2 for mitochondrial and cytosolic enzyme, respectively. The enzyme was inhibited by erythro-9-(2-hydroxy-3-nonvl)adenine and 2'-deoxycoformycin with A'i 4.4 X 10~7 M and 3.2 x 10 M for mitochondrial ADA and 4.9 X I0~7 M 2.8 x 10"7 M for cytosolic ADA. Among the natural nucleoside and deoxynucleotide derivatives tested, deoxyGTPand UTP inhibited only cytosolic adenosine deaminase by 60% and 40%, respectively.
This aim of the study was to determine the activity of ADA serum and isoenzyme in feline retroviral infections. The study involved 6 FeLV seropositive, 4 FIV seropositive and 10 healthy seronegative cats aged between 1-8 years. Haematological, serum enzyme acivity (AST; ALT; ALP; GGT) as well as ADA serum and isoenzyme activity were determined in all the cats. Haematological parameters were within the normal range except for the platelet count in FIV seropositive cats (p<0.05). Serum enzyme activity was unchanged except for the AST concentration in FeLV and FIV seropositive cats compared to the healthy subjects. ADA serum and ADA1 concentrations were lower in the seropositive group than in the seronegative group. However, the decrease in ADA serum and ADA1 concentrations in FIV seropositive cats (p<0.01) was more significant than that of FeLV seropositive cats (p<0.05). In conclusion, decreased ADA and ADA1 activity in feline retrovirus infections may be significant.
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