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

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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)

We have used an alkaline phosphatase protection assay to investigate the interaction of the trp repressor with its operator sequence. The assay is based on the principle that the trp repressor will protect a terminally 5'-32P-labeled operator DNA fragment from attack by alkaline phosphatase. The optimal oligonucleotide for investigating the trp repressor/operator interaction extends two base pairs from each end of the genetically defined target sequence predicted by in vivo studies [Bass et al. (1987) Genes Dev. 1, 565-572]. The assay works well over a 10,000-fold range of protein/DNA affinity and is used to show that the corepressor, L-tryptophan, causes the liganded repressor to bind a 20 base pair trp operator duplex 6400 times more strongly than the unliganded aporepressor. The affinity of the trp repressor for operators containing symmetrical mutations was interpreted in terms of the trp repressor/operator crystal structure as follows: (1) Direct hydrogen bonds with the functional groups of G-9 of the trp operator and the side chain of Arg 69 of the trp repressor contribute to DNA-binding specificity. (2) G-6 of the trp operator is critical for DNA-binding specificity probably because of the two water-mediated hydrogen bonds between its functional groups and the N-terminus of the trp repressor's E-helix. (3) Sequence-dependent aspects of the trp operator's conformation help stabilize the trp repressor/operator complex.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:An alkaline phosphatase protection assay to investigate trp repressor/operator interactions. 198 82

Two alternative procedures are described for the quantitative determination of phosphatidylcholine in a flow-injection system utilizing immobilized enzymes. Phospholipase C from Bacillus cereus and phospholipase D from cabbage were covalently bound to the surface of controlled-pore glass beads and the enzyme-derivatized beads were packed in small columns. In the first procedure, the phospholipase C column was connected with a second column containing coimmobilized alkaline phosphatase and choline oxidase. In the alternative procedure, the column packed with immobilized phospholipase D was connected with a column packed with immobilized choline oxidase. The hydrogen peroxide produced through the action of choline oxidase in both flow-injection systems was detected amperometrically. Both procedures are suitable for an accurate and rapid quantitation of phosphatidylcholine. The sensitivity of the method based on phospholipase C and alkaline phosphatase is higher than that using phospholipase D. Quantitation of phosphatidylcholine at the nanomole level can be easily obtained using the first method.
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PMID:Determination of phosphatidylcholine in a flow injection system using immobilized enzyme reactors. 220 Mar 5

Cells of Cryptococcus neoformans grown on xanthine or urate as the sole sources of nitrogen produced numerous, single membrane-bound organelles, deemed to be microbodies. Electron images of these structures showed positive cytochemical staining for catalase and alpha-hydroxy acid oxidase, known marker enzyme activities for microbodies. Microbodies in xanthine and urate-grown cells were cytochemically reactive for the presence of the hydrogen peroxide-producing xanthine and urate oxidases. Molybdenum and phosphorus (elements associated with the cofactor common to nitrogen scavenging enzymes) were detected in the substrate-induced microbodies by X-ray dispersive microanalysis. The single limiting membrane of the substrate-induced microbody was stained by a modified Gomori reaction for the presence of alkaline phosphatase, thereby suggesting the participation of this enzymic activity in the events associated with microbody chemistry.
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PMID:Electron cytochemical studies of Cryptococcus neoformans grown on uric acid and related sources of nitrogen. 221 35

The purpose of this study was to investigate the adaptability of long bones of young adult rats to the stress of chronic aquatic exercise. Twenty-eight female Sabra rats (12 weeks old) were randomly assigned to two groups and treatments: exercise (14 rats) and sedentary control (14 rats) matched for age and weight. Exercised animals were trained to swim in a water bath (35 degrees +/- 1 degree C, 1 hour daily 5 times a week) for 12 weeks loaded with lead weights on their tails (2% of their body weight) (BW). At the end of the training period following blood sampling for alkaline phosphatase, all rats were sacrificed and the humeri and tibiae bones were removed for the following measurements: bone morphometry, bone water compartmentalization, bone density (BD), bone mineral content (BMC), and bone ions content (Ca, Pi, Mg, Zn). The results indicate that exercise did not significantly affect the animals' body weight, bone volume, or length and diameters. However, bone hydration properties, BD, bone mass, and mineralization revealed significant differences between swim-trained rats and controls (P less than 0.05). Longitudinal (R1) measurement was higher by 43% for both humerus and tibia, and Transverse (R2) relaxation rates of hydrogen proton were higher by 117 and 76% for humerus and tibia, respectively; fraction of bound water was higher by 36 and 46% for humerus and tibia, respectively. BD, bone weight, and ash were higher by 13%. BMC and bone ions content were higher by 10%, and alkaline phosphatase was higher by 67%.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:The effect of swimming on bone modeling and composition in young adult rats. 222 93

This prospective study was undertaken in patients scheduled for gastrectomy for peptic ulcer disease to determine the effect of partial gastrectomy with either Roux-en-Y (n = 11) or Billroth II anastomosis (n = 11) on the function of the small intestine. Patients were studied before and at 6 months (blood and small-intestinal function tests) and at 24 months (blood tests) postoperatively. Median postoperative body weights at 6 months (70.5 kg; p less than 0.01) and 12 months (70.3 kg; NS) were lower than preoperatively (73.0 kg). Haemoglobin concentrations at 6 months (8.9 mM; p less than 0.01) and at 24 months (9.1 mM; p less than 0.05) were also significantly reduced compared with the preoperative value (9.5 mM). However, neither at 6 nor at 24 months postoperatively were there significant changes for serum iron, iron saturation, folic acid, vitamin B12, protein, albumin, alkaline phosphatase, and calcium concentrations. Whereas no significant deterioration of the absorption of D-xylose and vitamin B12 or of faecal fat excretion was observed, the orocoecal transit time was significantly shortened from 98 to 50 min (p less than 0.01), the expiratory hydrogen excretion after a 50-g oral glucose load was significantly increased from 8 to 54 ppm (p less than 0.01), as was indicanuria from 257 to 368 mumol/24 h (p less than 0.01). Apart from a lower serum iron concentration and iron saturation index in the Roux-en-Y patients 6 months postoperatively (p less than 0.05), no significant differences between the two types of anastomosis were observed. It is therefore concluded that both in patients with Roux-en-Y and in those with Billroth II anastomosis most abnormalities observed after gastrectomy are secondary to an accelerated small-intestinal transit.
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PMID:Effect of gastrectomy with either Roux-en-Y or Billroth II anastomosis on small-intestinal function. 230 15

A highly sensitive chemiluminescent assay for NAD(P)H have been developed. The principle of the method is as follows; NAD(P)H reduces molecular oxygen to superoxide anion (O2-) and hydrogen peroxide (H2O2) in the presence of 1-methoxy-5-methylphenazinium methyl sulphate (1-MPMS) as electron mediator. The produced O2- and H2O2 can be measured by chemiluminescent reaction using isoluminol (IL) and microperoxidase (m-POD). A linear relationship between chemiluminescence intensity and NAD(P)H concentration (log/log) was obtained ranged from 10(-9) mol/l to 10(-5) mol/l. This chemiluminescent reaction has been coupled to the assay of glucose-6-phosphate dehydrogenase (G6PDH), beta-D-galactosidase (beta-Gal) and alkaline phosphatase (ALP). The detection limits of G6PDH, beta-Gal and ALP were 10(-18) mol, 10(-20) mol and 10(-18) mol per assay, respectively. The chemiluminescent assay of these enzymes applied to chemiluminescent enzyme immunoassay for 17 alpha-hydroxy-progesterone and DNA hybridization assay using these enzymes as label.
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PMID:Chemiluminescent assay of various enzyme activities and its application to enzyme immunoassays. 250 34

Immunoblotting techniques are widely used for detection of antigen immobilized on nitrocellulose membranes. There are many immunolabeling methods and staining methods available to disclose the presence of antigen in such techniques. Five common staining methods each for alkaline phosphatase and horseradish peroxidase were examined. The staining methods with the highest sensitivity and the lowest background were selected for studies comparing five immunological labeling methods using human IgG as a model antigen. Results were evaluated on the basis of the least amount of detectable antigen and background staining. The most sensitive dot-blot method was then tested for its applicability to Western blots. For both dot-blots and Western blots, the immunoalkaline phosphatase methods are more sensitive than the corresponding immunoperoxidase methods. The use of biotinylated secondary antibodies and an avidin-enzyme conjugate is recommended. Disclosure of alkaline phosphate is best achieved with naphthol AS phosphate as substrate and fast blue BB as chromogen. Peroxidase is best stained using H2O2 and diaminobenzidine (DAB). Potential endogenous enzyme activities are demonstrable by blotting methods but can be inhibited by including levamisole in the disclosure reaction medium for calf intestinal alkaline phosphatase indicators, or by incubation of blots with sodium azide and hydrogen peroxide before immunolabeling when using horseradish peroxidase indicators.
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PMID:Assessment of a method for immunochemical detection of antigen on nitrocellulose membranes. 253 57

The liver, heart, lungs, and stomach of rats exposed to hydrogen fluoride were studied. Histological examination showed partial liver necrosis and emphysema. Using histochemical methods the effect of fluorine ions was found in: a reduction of the activity of succinic and beta-hydroxybutyric dehydrogenases in the liver, heart muscle, superficial and glandular epithelium cells, and in lamina propria of the gastric mucosa; an increase in the activity of lactate dehydrogenase in liver cells; an increase in the activity of acid phosphatase in the liver, heart muscle, bronchus epithelium, bronchioli, and interalveolar septum cells; an increase in the activity of alkaline phosphatase in the liver, lungs, and heart muscle connective tissue, and in all gastric epithelium cells. The results obtained mainly point to the inhibition of oxidative metabolism by fluoride ions.
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PMID:Studies on the toxicology of fluorine compounds. I. Histological and histochemical investigations on the liver, heart, lungs, and stomach of rats exposed to hydrogen fluoride. 261 87

Since they are found to be increased in lesions of acute necrotic ulcerative gingivitis or marginal periodontitis, agents for these diseases. In the present study, 38 pure cultured strains were obtained as a result of isolation and culture of samples collected from lesions of marginal periodontitis (periodontal pokets), and the biological and biochemical characteristics of these strains were investigated. 1) Light microscopy (including dark-field microscopy) and transmission electron microscopy (negative staining) were used for observation of the morphology and cellular structure of the strains. The cells had a spiral shape, and showed active movement. Based on the above findings the cultured strains were all confirmed to be spirochetes of small to medium size, being 0.08-0.24 micron in width. 2) Growth and motility of the strains were investigated on various types of culture medium. Intense growth and movement were noted in strains cultured in bovine liver exudate medium containing horse serum (pH 7.2) at 37 degrees C under anaerobic conditions produced by the evacuation-replacement method (95% N2, 5% CO2) for 3-7 days after inoculation. 3) Thirty-five strains were positive for indole production and decomposition of urea, mucin, hippuric acid and esculin. Production of hydrogen sulfied was observed in 31 strains. In decomposition tests for 17 carbohydrates, 17 strains were positive for galactose and 14 strains were positive for glucose, while 11 strains were positive for dextrin and 10 strains for fructose upon decomposition of soluble starch. Other carbohydrates were also decomposed by a few strains. 4) In an investigation of the production of alcohol and lower fatty acids, among the metabolic products detected by gas chromatography, a large amount of acetic acid and small amounts of ethanol, lactic acid, propionic acid, pyruvic acid were observed. 5) The results of enzyme activity tests using an API ZYM system indicated relatively high activities of esterase, esterase-lipase, alpha-glucosidase, alkaline phosphatase, trypsin and acid phosphatase.
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PMID:[Biological and biochemical characteristics of the oral spirochetes isolated from the focus of marginal periodontitis]. 276 48

Neutrophils and band forms from patients with acute myeloid leukemia and myelodysplastic syndrome were stained for the presence of myeloperoxidase using a cytochemical method (diaminobenzidine/hydrogen peroxide) and the alkaline phosphatase--anti-alkaline phosphatase immunocytochemical procedure (using monoclonal anti-myeloperoxidase). Neutrophils and bands were also stained for elastase and lactoferrin using monoclonal and polyclonal antibodies, respectively. Subpopulations of neutrophils and bands from cases of acute myeloid leukemia and myelodysplasia exhibited a qualitative and/or quantitative deficiency in myeloperoxidase. In addition, a quantitative decrease in elastase and/or lactoferrin staining was detected. Thus, neutrophils and bands from patients with acute myeloid leukemia and myelodysplastic syndrome have a defect in one or more of the constituents of primary and/or secondary granules. These defects are consistent with the view that abnormal neutrophils and bands are derived from a malignant clone of myeloid precursor cells.
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PMID:Abnormal neutrophils in acute myeloid leukemia and myelodysplastic syndrome. 283 2


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