Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A method using p-benzoquinone for coupling antigens and antibodies to enzymes and erythrocytes is described. The method involves the treatment of proteins (or polysaccharides) at pH 6 or 7 with an excess of p-benzoquinone. After removal of the unreacted reagent by gel filtration, the "activated" proteins were coupled at pH 8-9 with enzymes or erythrocytes. Biological activities of the proteins were not substantially modified by this treatment since 80-100% of the antigen binding capacity was found to be preserved in p-benzoquinone treated antibodies or Fab fragments. Anti-Ig antibodies (or Fab) were coupled by this procedure to peroxidase, alkaline phosphatase, lactoperoxidase, glucose oxidase and beta-galactosidase, and the conjugates obtained were found to be highly effective in detecting intracellular Ig by immunohistochemical techniques. Erythrocytes coated with sheep anti-mouse Ig antibody or Fab were used to titrate by passive hemagglutination serum Ig. The same erythrocytes were employed to detect by plaque assay mouse Ig secreting cells. Erythrocytes coated with peroxidase, alkaline phosphatase, bovine serum albumin, ribonuclease, Salmonella polysaccharide (B 27 +) and pneumoccocal polysaccharide SIII were employed to titrate serum antibody by passive hemagglutination and hemolysis and to detect mouse antibody secreting cells by plaque assay. All the antigens and antibodies coated erythrocytes prepared gave highly satisfactory and reproducible results.
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PMID:A new method using p-benzoquinone for coupling antigens and antibodies to marker substances. 0 79

A new method for quantifying class-specific antibodies is presented. The method has been named Diffusion-In-Gel-Enzyme-Linked-ImmunoSorbentAssay (DIG-ELISA), and is briefly as follows. Antiserum ia allowed to diffuse from wells in a gel layered over an antigen-coated plastic surface. The gel is then removed and the preparation is incubated with enzyme-conjugated anti-immunoglobulin. The enzyme is then visualised in situ by a colour reaction produced by pouring a substrate-containing gel over the plastic surface. Bovine serum albumin and rabbit-anti-BSA were used as a model system, and horseradish peroxidase or alkaline phosphatase as enzymes for visualization.
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PMID:Diffusion in gel-enzyme linked immunosorbent assay (DIG-ELISA): a simple method for quantitation of class-specific antibodies. 11 55

Biochemical changes were monitored during fat infusion as part of an evaluation of the toxicity that accompanies the intravenous administration of a new soybean oil emulsion in dogs. Beagles were given FE-S15 at 9 and 4 g/kg/day for 28 days via a central venous catheter. The total serum lipid, triglyceride, and phospholipid concentrations of the animals receiving fat in high doses increased 3 to 4 times in comparison to that of the control group; cholesterol increased 5 times. The rise of serum lipid fractions was proportionally smaller in animals receiving the low dose fat infusion. All values returned to the control range 2 wks after termination of the infusion. The serum protein level fell from 6.5 to 5.1 g/dl in animals given 9 g/kg/day fat while animals receiving 4 g/kg/day had a significant increase to 8.4 g/dl (day 27). In comparison with the control groups, the animals receiving low dose fat infusion also had a significant rise in serum albumin and, to a lesser degree, in alpha and beta globulins. A rise in alkaline phosphatase activity was observed in all dogs receiving fat infusion, but was significantly different from the controls only in animals on high dose fat infusion. None of the other monitored biochemical serum values were affected by the administered fat and there was no evidence of disturbance in liver function.
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PMID:Studies of the toxicity of an intravenous fat emulsion. II. Blood chemical changes after administration of a soybean oil (FE-S15) in beagles. 11 24

This paper describes an Enzyme Linked Immunosorbent Assay (ELISA) for detecting IgG sensitized erythrocytes utilizing a commercially available anti-human IgG conjugated with alkaline phosphatase. Erythrocyte hemolysis in the assay was minimized by dissolving the p-nitrophenyl phosphate substrate in a carbonate-bicarbonate buffer. Nonspecific absorption of the enzyme conjugate to erythrocytes and glassware was reduced by adding 1% bovine serum albumin to wash solutions. Assay sensitivity was increased with greater concentrations of enzyme conjugate and erythrocytes in the incubation stage. The sensitivity of the described ELISA procedure is approximately equal to that of the standard antiglobulin test. Some possible future applications of ELISA in the blood bank are discussed.
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PMID:An enzyme linked immunosorbent assay (ELISA) for detecting IgG sensitized erythrocytes. 11 58

The catalytic subunit of rabbit skeletal muscle cyclic AMP-dependent protein kinase I can catalyze self-phosphorylation. The autophosphorylation reaction uses ATP as the phosphoryl donor, requires Mg2+, and is inhibited by polyarginine. Prior treatment of the catalytic subunit with Escherichia coli alkaline phosphatase in the presence of bovine serum albumin greatly enhances the autophosphorylation of the subunit. The protein-bound phosphate is stable in acid but labile in base. Incubation of the 32P-labeled phosphoenzyme with histones led neither to the phosphorylation of histones nor to a loss of radioactivity from the phosphoenzyme. The results suggest that the phosphoenzyme does not represent an intermediate of the phosphotransferase reaction.
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PMID:Autophosphorylation of rabbit skeletal muscle cyclic AMP-dependent protein kinase I catalytic subunit. 21 18

Calcium and vitamin D metabolism were evaluated in 5 adult epileptic patients before and during treatment with phenytoin. Significant decreases occurred in serum concentrations of calcium, albumin, and 25-hydroxy-cholecalciferol. The decreases in serum calcium paralleled those in serum albumin. Significant increases occurred in serum alkaline phosphatase and 1 alpha, 25-dihydroxycholecalciferol, in urinary hydroxyproline, and in the fractional gastrointestinal absorption of calcium. Urinary cyclic adenosine monophosphate and serum parathyroid hormone did not change. The results suggest that the bone disease resulting from phenytoin therapy may be associated with a deficiency of 25-hydroxycholecalciferol and not of 1 alpha, 25-dihydroxycholecalciferol, and that reduced gastrointestinal absorption of calcium or changes in parathyroid function may not be necessary for the development of bone disease.
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PMID:Effect of phenytoin on bone and vitamin D metabolism. 22 Sep 3

Two subpopulations (F4 and F6) of guinea-pig thymocytes were separated by using bovine serum albumin gradient centrifugation. The majority of F4 thymocytes were weakly alkaline phosphatase (AP) positive cells, while most of F6 thymocytes were strongly AP positive. The significant difference between their AP activities was confirmed biochemically. In the ultracytochemical study the majority of unfractionated and F6 thymocytes were light and AP positive, whereas F4 thymocytes were mostly dark, weakly AP positive cells. F4 thymocytes responded well to PHA and Con A, while F6 thymocytes failed to respond to these mitogens. The subpopulations did not differ in their homing properties. These findings indicate that strongly AP-positive cells are immature thymocytes and weakly AP-positive cells represent a more mature subpopulation of thymocytes. A hypothetical scheme for differentiation of guinea-pig thymocytes is presented.
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PMID:Two subpopulations of guinea-pig thymocytes with different maturation stages. 30 22

The levels of membrane alkaline phosphatase have been measured on different lymphocyte fractions from human peripheral blood separated on bovine serum albumin discontinuous gradients. The peak in enzyme activity was observed in a non-T-, non-B-cell fraction, rich in "null" lymphocytes; the lowest values were found in the fraction with the highest proportion of T-cells.
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PMID:Alkaline phosphatase in human lymphocyte subpopulations. 31 Dec 97

As part of a double-blind, randomized, controlled trial to evaluate the effect of colchicine on liver cirrhosis, 43 cirrhotic patients were assigned to either a placebo (20 patients) or a colchicine (23 patients) treatment group. Colchicine 1 mg and an indistinguishable placebo were administered orally on a daily dose 5 days a week. In the colchicine group, 12 were males and 11 females, while in the control group 13 were males and 7 females. The time elapsed between diagnosis and inclusion in the study was 14.1 mo for the controls and 14.5 mo for the patients on colchicine. Mortality related to the liver disease occurred in 4 patients on colchicine and 8 patients on placebo. Although the probability of surviving in the colchicine group was greater than that of the placebo, the difference did not reach statistically significant levels. Of the colchicine-treated patients, in three a remarkable decrease in liver fibrosis was observed in serial biopsies. In two other patients, carcinoma of the liver developed. Six of the survivors on colchicine have improved clinically, noticing disappearance of ascites and edema, as well as a decrease in the size of the spleen. All the survivors on placebo continue to show clinical deterioration. In contrast to the usual drop of serum albumin seen in the cirrhotic patients, those receiving colchicine increased and maintained their serum albumin levels throughout the study. Serum proline values were elevated only in the alcohol cirrhotic patients. Serum alkaline phosphatase increased only in those patients receiving colchicine. The results indicate that in some cases, liver fibrosis could be modified by treatment with antifibrotic drugs. The use of colchicine at present should remain within controlled studies.
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PMID:Treatment of cirrhosis with colchicine. A double-blind randomized trial. 37 54

The overall transport of bile salts across the hepatocyte is characterized as a carrier-mediated process whose rate-limiting step is biliary secretion. Specific bile salt binding proteins have been identified in liver surface membrane fractions and were postulated to represent the initial interaction in bile salt translocation across both the sinusoidal and canalicular membranes. To test this hypothesis, cycloheximide was administered to rats to inhibit hepatic protein synthesis. 16 h after cycloheximide administration [14C]leucine incorporation into hepatic protein was inhibited by 93% at 1 h and 47% at 12 h. However, values of liver function tests were not increased, although serum albumin, serum alanine amino-transferase, and alkaline phosphatase were significantly decreased. Light and electron microscopy did not demonstrate necrosis or fat accumulation. The latter demonstrated minimal disorganization of rough endoplasmic reticulum and occasional lamellar whorls. 16 h after cycloheximide administration bile salt independent bile flow, basal bile salt excretion, and basal bile flow were unaltered, but the maximum bile salt transport capacity was reduced to 62% of control and 24 h later to 38%. Decreased bile salt transport was reversible, for it returned to control values after 48 h, when hepatic protein synthesis was also normal. Maximum bromosulfophthalein (BSP) transport, on the other hand, was reduced after 16 h to only 85% of control. Both bile salt and BPS maximum transport capacities decreased with time during inhibition of protein synthesis, apparently following first order kinetics. It was estimated that their half-lives are 20 h for bile salt transport and 55 h for BSP transport. These different turnover rates suggest that cycloheximide does not decrease active transport through generalized hepatic dysfunction or alteration of high energy sources possibly required for transport. The maximum number of [14C]cholic acid binding sites in liver surface membrane fractions was determined by an ultrafiltration assay. They were reduced to 68% of control after 16 h of cycloheximide and to 25% after 24 h. This reduction in the number of binding sites is apparently selective, for the activities of the liver surface membrane enzymes (Na+-K+)ATPase, Mg++-ATPase, and 5'-nucleotidase were not significantly changed. The associated alterations in bile salt transport and the maximum number of binding sites after cycloheximide administration suggests that these receptors may be the bile salt carriers.
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PMID:Regulation of hepatic transport of bile salt. Effect of protein synthesis inhibition on excretion of bile salts and their binding to liver surface membrane fractions. 43 30


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