Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have developed an enzyme immunoassay (ELISA) for the quantification of the schistosome circulating cathodic antigen (CCA), a glycoprotein associated with the syncitium lining the gut of the parasite. A mouse monoclonal antibody of IgG3 isotype was used as coating (antigen-capture) antibody, while a biotinylated mouse monoclonal IgM was used as second (antigen-detecting) antibody. Streptavidin-alkaline phosphatase was used as enzyme label. The lower detection limit of the assay was 1.0 ng of the trichloroacetic acid soluble fraction of adult worm antigen (AWA-TCA) per ml, which corresponds to approximately 0.2 ng CCA per ml. The ELISA showed a linear range from 1.0 to 62.5 ng AWA-TCA per ml. Serum and urine samples of 16 individuals infected with Schistosoma mansoni (egg counts ranging from 5 to 4820 eggs per gram of faeces) were tested in the assay. Antigen titres ranged from less than 4-8192. This assay represents a considerable advantage in diagnosis of Schistosoma infections as it allows the detection and quantification of CCA in serum and urine in even lightly infected individuals.
...
PMID:Detection of the schistosome circulating cathodic antigen by enzyme immunoassay using biotinylated monoclonal antibodies. 212 84

We report the development of a time-resolved immunofluorometric assay (TR-IFMA) for the quantitative determination of the schistosome circulating anodic antigen (CAA). A mouse monoclonal antibody (line 120-1B10-A), recognizing a repetitive epitope on CAA, was used as both antigen-capture antibody and as Europium-labelled antigen-detecting antibody. The lower detection limit of the assay was 20 pg of trichloroacetic acid soluble fraction of adult worm antigen (AWA-TCA) per ml, with a nearly linear measuring range from 20 pg to 130 ng AWA-TCA per ml. The TR-IFMA was compared with an enzyme-linked immunosorbent assay (ELISA), in which the same monoclonal antibody was used as antigen-capture antibody and as alkaline phosphatase-conjugated antibody. The lower detection limit of the TR-IFMA was tenfold lower than that of the ELISA, while the linear range of the TR-IFMA exceeded that of the ELISA one hundred-fold. Serum samples of 80 Burundese individuals infected with Schistosoma mansoni (egg counts ranging from 4 to 2583 eggs per gram of faeces) were tested in both assays. Antigen concentrations in serum of individuals infected with S. mansoni ranged from 0-500 ng AWA-TCA per ml. The correlation between antigen levels measured by TR-IFMA and ELISA was good: Spearman's p = 0.92. Whereas in the ELISA the samples had to be titrated, the wide linear range of the TR-IFMA allowed the assay to be performed at a single serum dilution, at which an exact estimation of the antigen concentration was possible.
...
PMID:Time-resolved immunofluorometric assay (TR-IFMA) for the detection of the schistosome circulating anodic antigen. 251 32

A patient with pyoderma gangrenosum without associated disease was studied. Routine investigations showed several abnormalities. High ESR, high alkaline phosphatase and glutamyl transferase (gamma-GT) levels, low iron and iron binding capacity, altered protein spectrum, presence of Staphylococcus aureus and group G hemolytic streptococci in ulcer culture, higher than normal antistreptolysin titers in the serum, and perivascular infiltration in the skin. Biochemical investigations aimed at finding any excessive hydrolytic activity did not reveal the presence of neutral proteases in circulation leaked out from PMN-leukocytes or elsewhere. Lysozyme levels were higher than normal, amylase and lipase levels were normal and 5' nucleotidase levels were below normal range. TCA-soluble polypeptides were present in the serum at levels two times higher than those in normal individuals. Immunochemical investigations showed the absence of immune complexes in the serum but presence of high amounts of C-reactive protein. Total complement activity was higher than normal and so was C3c level. Clq, C4, and C3d levels were within normal range. Biologic studies showed the presence of a factor in patient serum that made guinea pig skin hard, painful, erythematous, and eventually hairless, but not necrotic. A similar factor was either absent in normal serum or present in very low concentration. After salazopyrine treatment, all the above mentioned abnormalities corrected except that 5' nucleotidase activity remained slightly lower than normal, alkaline phosphatase levels remained slightly higher than normal, and C-reactive protein levels remained very high, though lower than those during intense disease activity.
...
PMID:Immunologic and biochemical studies on a patient with pyoderma gangrenosum. 614 8

The effects of l and d-, p-bromotetramisole (pBTM) on alkaline phosphatase were studied in relation to 45Ca2+, 32phosphate and 3H-thymidine uptakes in non-mineralizing second (M2) and mineralizing first (M1) maxillary hamster molar tooth germs under the conditions of organ culture. At the concentration used in culture (10(-3)M), l-pBTM completely inhibited alkaline phosphatase activity in tooth germ homogenates. About 30% of the enzyme activity was inhibited by d-pBTM at the same concentration. In culture, there were no significant differences between the effects of l and d-pBTM isomers on all the parameters measured. In the non-mineralized M2 molars, l and d-pBTM significantly reduced both TCA-soluble and TCA-insoluble 32phosphate uptakes but not 45Ca2+. However, 3H-thymidine uptake was also significantly decreased. In M1 molars, the pBTM isomers significantly reduced the uptake of TCA-soluble 32phosphate and 45Ca2+ but not TCA-insoluble 32phosphate. Ouabain, a specific inhibitor of Na+-K+-ATPase (but not alkaline phosphatase), also significantly reduced 3H-thymidine uptake to the same extent as the pBTM isomers in the non-mineralizing M2 molars, but it did not significantly affect either 32phosphate (TCA-soluble and TCA-insoluble) or 45Ca2+ uptake. Although this inhibitor significantly reduced both 45Ca2+ and TCA-soluble 32phosphate uptake in the mineralizing M1 molars, this effect was much less dramatic than was the case with the pBTM isomers. The reduced 45Ca2+ uptake in the M1 molars is probably a consequence of reduced mineralization since in the non-mineralizing M2 molars calcium uptake was not significantly affected by the pBTM isomers.(ABSTRACT TRUNCATED AT 250 WORDS)
...
PMID:Studies of alkaline phosphatase inhibition by p-bromotetramisole in non-mineralizing and mineralizing neonatal hamster tooth germs in vitro. 658 Nov 67

Phosphate binding by rat renal brush border membranes occurs on a single protein, as visualized by SDS polyacrylamide gel electrophoresis. The same protein can also be specifically labelled by gamma-32P ATP at 0 degree C or in the absence of magnesium. The phosphate binding protein co-migrates with monomers of two alkaline phosphatase activity bands previously localized on acrylamide gel. Measurement of binding by TCA precipitation, ion-exchange chromatography and dialysis gave an average of 31.1 +/- 5.7 pmol phosphate bound/mg protein. Alkaline phosphatase would then represent 0.23% of total brush border membrane protein. Maximal binding activity is obtained at pH 6.5, but when membranes are phosphorylated at pH 6.5 and the pH increased to 9.4, 50% of the bound radioactivity is released. The binding of phosphate to this protein presents two different apparent Km: one at 40 microM for low and one at 390 microM for high substrate concentrations. The membrane bound phosphate is readily exchangeable with phosphate in the medium. Phosphate binding and phosphate release are complete within 5 s. Alkaline phosphatase substrates and EDTA are potent inhibitors of phosphate binding and produce over 90% inhibition. Characteristics of phosphate binding for kidney membrane bound alkaline phosphatase seem very similar to the soluble form of the enzyme from various sources.
...
PMID:Characterization of phosphate binding by alkaline phosphatase in rat kidney brush border membrane. 663 81

Chicken skeletal alkaline phosphatase is subject to competitive inhibitions by vanadate (Ki = 0.38 mM in carbonate, Ki = 0.08 mM in Tris, both at pH 7.4) and phenylphosphonate (Ki = 15 mM in carbonate, Ki = 1.3 mM in Tris, both at pH 7.4), and uncompetitive inhibition by levamisole (Ki = 0.08 mM in carbonate, Ki = 0.10 mM in Tris, both at pH 7.4). The competitive inhibitors were more effective in Tris buffer because nonreactive ternary complexes were formed between alkaline phosphatase, the inhibitor and Tris. The effects of vanadate, phenylphosphonate and levamisole on the proliferation of embryonic chick calvarial cells in vitro were biphasic. Low doses of each agent stimulated 3H-thymidine incorporation into TCA-insoluble material; higher doses were inhibitory. Neither effect could be attributed to inhibition of alkaline phosphatase activity (e.g. 20 microM vanadate should inhibit alkaline phosphatase by 3% but stimulated cell proliferation by 187%; 50 microM vanadate should inhibit alkaline phosphatase by 7% but inhibited 3H-thymidine incorporation by 90%). None of the alkaline phosphatase inhibitors tested affected the cellular concentration of the enzyme during the 24-hour incubation. These studies indicate that alkaline phosphatase inhibitors can have nonspecific effects on bone cells in culture, and that for cells in the osteoblast cell line, an inhibition of alkaline phosphatase activity is not consistently related to a decrease in cell proliferation.
...
PMID:Effect of skeletal alkaline phosphatase inhibitors on bone cell proliferation in vitro. 692 63

With Hyroxylapatite purified preparations and BACH (biotin aminocapryl hydrazide) biotinylated McAbs, 274-2H10 and 273-2H1, recognizing different egg-associated epitopes, biotin-avidin (BA) involved alkaline phosphatase (AP) ELISA with detecting sensitivities reaching nanogram levels (10(-9), were set up. The detectable limit for crude preparations of Schistosoma japonicum SEAJ-TCA in 2H10-ELISA achieved 1. 0.3. 2 ng/ml, in which only S. japonicum specific egg antigens were efficiently detected, whereas with 2H1-ELISA, which could detect SEA-TCA of both S. japonicum and S. mansoni species, an end point of detecting 3.2 ng/ml was obtained. Repeated tests with human serum groups revealed very significant differences of extinction OD readings between patients and normal individuals. For detection combinations, a previously established anti-CAA homologous AP-ELISA system was parallelly used for gut-associated antigenemia determinations. Taking the mean extinction OD reading of a parallel normal serum group plus 3 SD as corresponding cut off values, 3 patient groups (n = 82, 52, 39) from different areas of transmission intensity were subjected to accumulating determinations for egg- and gut-associated antigenemia. Improved detectabilities to variable extent were achieved in either of the 2 or 3 combinations. The study thus demonstrated that the diagnostic efficiency for human schistosomiasis could be improved by multi-epitope detections for more than one target molecule using corresponding McAbs, especially in areas where the transmission intensity of the disease is comparatively lower.
...
PMID:[A preliminary report on diagnostic complementarity of gut-associated and egg-associated antigenemia in schistosomiasis japonica]. 754 May 18

The effect of DL alpha-lipoic acid on the nephrotoxic potential of gentamicin was examined. Intraperitoneal injection of gentamicin (100 mg/kg/day) to rats resulted in decreased activity of the glycolytic enzymes-hexokinase, phosphoglucoisomerase, aldolase and lactate dehydrogenase. The two gluconeogenic enzymes--glucose-6-phosphatase and fructose-1,6-diphosphatase, the transmembrane enzymes namely the Na+, K(+)-ATPase, Ca(2+)-ATPase, Mg(2+)-ATPase and the brushborder enzyme alkaline phosphatase, also showed decreased activities. This decrease in the activities of ATPases and alkaline phosphatase suggests basolateral and brush border membrane damage. Decreased activity of the TCA cycle enzymes isocitrate dehydrogenase (ICDH), succinate dehydrogenase (SDH) and malate dehydrogenase (MDH), suggests a loss in mitochondrial integrity. These biochemical disturbances were effectively counteracted by lipoic acid administration. Lipoic acid administration by gastric intubation at two different concentrations (10 mg and 25 mg/kg/day) brought about an increase in the activity of the glycolytic enzymes, ATPases and the TCA cycle enzymes. The gluconeogenic enzymes however showed a further decrease in their activities at both the concentrations of lipoic acid administered. These observations shed light on the nephroprotective action of lipoic acid against experimental aminoglycoside toxicity and the protection afforded at 25 mg/kg/day of lipoic acid was noted to be higher than that at 10 mg level.
...
PMID:Role of DL alpha-lipoic acid in gentamicin induced nephrotoxicity. 765 73

There is no satisfactory explanation for the cholinergic deficit characteristic of Alzheimer's disease. We have performed a series of experiments which demonstrate that (a) an inhibitor of cytosolic human brain choline acetyltransferase is present in the cytosol of Alzheimer brain tissue, (b) human brain cytosolic choline acetyltransferase activity is inhibited by phospho-L-serine in a competitive manner. Cytosol was prepared from human forebrain or amygdala and the Km for choline and acetyl CoA of the choline acetyltransferase were 750 microM and 12.5 microM, respectively. Phospho-L-serine was found to be a competitive inhibitor of this enzyme with respect to choline but not with respect to acetyl CoA with a Ki of 750 microM for the human forebrain and 3 mM for human amygdala. These concentrations of phospho-L-serine are present in brain tissue at early stages of Alzheimer's disease. Several other phosphomonoesters and phosphodiesters that are increased in Alzheimer's disease were either less inhibitory or without effect. The addition of heat denatured and non-heat denatured cytosol from Alzheimers forebrain inhibited the choline acetyltransferase activity present in control human brain cytosol. The inhibitory activity of the Alzheimers cytosol was retained in TCA deproteinized samples and removed by dialysis or by alkaline phosphatase treatment. Dialysis of the cytosol increased the choline acetyltransferase activity of 5 of 8 Alzheimer's disease samples from 21 to 118% with p values of < 0.025 or < 0.001, respectively. These observations provide evidence that an endogenous non-proteinaceous, dialyzable, phosphomonoester, present in Alzheimers brain inhibits the choline acetyltransferase of both control and Alzheimers brain.(ABSTRACT TRUNCATED AT 250 WORDS)
...
PMID:Inhibition of cytosolic human forebrain choline acetyltransferase activity by phospho-L-serine: a phosphomonoester that accumulates during early stages of Alzheimer's disease. 836 18

The effects of 1,25-dihydroxyvitamin D3 [1,25(OH)2,D3], 1,25-dihydroxy-16ene-23yne-vitamin D3[1,25(OH)2-16ene-23yne-D3] (a synthetic analog) and retinoic acid on proliferation, protein synthesis, and alkaline phosphatase activity and mRNA were compared in two late-passage (P > 70) clonal rat osteoblastic cell lines (G2 and C12) in order to characterize variations in the basal and hormonally-regulated phenotypes. All agents inhibited proliferation (measured as cell number after 3 days of treatment) in late-passage (P > 70) G2 and C12 cells without inhibiting the rate of protein synthesis ([3H]leucine incorporation into TCA-precipitable protein) during the last 18 h of incubation. Basal and hormone-treated alkaline phosphatase activities were lower in late-passage G2 and C12 cells than those previously reported for early-passage G2 and C12 cells. 1,25(OH)2D3 and 1,25(OH)2-16ene-23yne-D3, up-regulated alkaline phosphatase activity in late-passage C12 cells and down-regulated it in late-passage G2 cells. The direction of these regulatory changes in late-passage cells was opposite to that reported for early passage of these clones, and changes were related to the levels of tissue-unspecific alkaline phosphatase mRNA normalized for actin mRNA. Effects of 1,25(OH)2D3 or 1,25(OH)2-16ene-23yne-D3 and retinoic acid were not additive, suggesting a competitive mechanism of action. It appears that increased sensitivity to the antiproliferative effects of regulatory hormones and defects in proliferation and specialization of the osteoblast are observed with increasing passage number in vitro in two model osteoblastic cell lines (G2 and C12).
...
PMID:Variation in 1,25-dihydroxyvitamin D3 regulation of proliferation and alkaline phosphatase activity in late-passage rat osteoblastic cell lines. 866 71


1 2 Next >>

---