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 recycling assay for alkaline phosphatase, based on its ability to hydrolyse NADP to NAD+, is presented. The product NAD+ is recycled in a coupled assay consisting of NADH regeneration and reduction of a nitroblue tetrazolium salt. This assay is 10-12 times more sensitive than the conventional assay. We demonstrate the role of energy poisons in transport of this protein into the periplasm by combining the improved detection with phase separation of the periplasmic and cytoplasmic alkaline phosphatase pools.
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PMID:A recycling assay for alkaline phosphatase applied to studies on its transport in E. coli K12. 269 14

We have developed a three-stage avidin-biotin alkaline phosphatase system (ABAP) for immunophenotyping acute and chronic leukaemias. Air-dried blood and bone marrow films or cytospins were fixed in acetone, methanol-acetone, or methanol-acetone-formalin. A primary monoclonal antibody layer was employed with a corresponding biotinylated anti-species second layer. The third stage was avidin-conjugated alkaline phosphatase and visualisation was achieved with Fast Red TR as diazo salt. This was compared using a limited range of primary antibodies with the alkaline phosphatase anti-alkaline phosphatase (APAAP) method, routinely performed as a five-stage technique. The ABAP technique provided a 2.5 h, reliable, three-stage immunophenotyping procedure without the need for amplification steps. A wider range of first layer monoclonals of any species could be utilised by using the appropriate secondary antibody. Preservation of cell morphology was very good. Smaller amounts of expensive monoclonal antibodies could be used compared to the quantity required with other slide techniques. This approach to immunophenotyping can consequently be employed by any general haematology laboratory to provide a sensitive and inexpensive service. The technique has been in routine diagnostic use for the last 2 years, and consistently good results have so far been achieved in a national external immunophenotyping quality assurance scheme.
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PMID:Rapid diagnosis of leukaemia: a three-stage immunophenotyping technique. 209 56

This paper reports on the use of alkaline phosphatase cytochemistry and combined conventional and confocal reflection and fluorescence scanning light microscopic modes in the study of human marrow stroma. It was found that the end product of the enzyme reaction using Napthol AS phosphate as substrate and Fast Blue BB as coupler reflected the 633 nm (red) light from a Helium-Neon laser. Serial optical sections suitable for 3-D reconstruction and selectively depicting the marrow reticulum cells could be obtained from thick glycol methacrylate sections reacted for Alkaline phosphatase. Furthermore, the yellow background of uncoupled diazonium salt over cytochemically unreactive structures in the same specimens and fields was used for imaging haemopoietic cell mass by operating the microscope at 488 nm (argon ion laser, blue-green). These methods may offer advantages in the investigation of the bone marrow stroma and its interplay with haemopoiesis and osteogenesis in normal and disease conditions.
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PMID:Alkaline phosphatase cytochemistry in confocal scanning light microscopy for imaging the bone marrow stroma. 273 May 14

Cytochemical methods for the demonstration of enzyme activities in blood and bone marrow cells were systematically improved by the addition of an inert polymer, polyvinyl alcohol (PVA), to the incubation medium and by using optimized reaction media. The methods investigated were tetrazolium salt methods for lactate, glucose-6-phosphate, succinate and glutamate dehydrogenase, the indoxyl-tetrazolium salt method for alkaline phosphatase, the diaminobenzidine method for peroxidase, and diazonium salt methods for chloroacetate esterase, beta-glucosaminidase, beta-glucuronidase, acid phosphatase, and dipeptidylpeptidase II and IV. PVA in the media preserved the morphology of cells very well and prevented leakage of large molecules such as enzymes from the cells. Therefore, fixation or long periods of air-drying prior to incubation leading to substantial loss of enzyme activity could be avoided. A brief period of drying (2 min at 37 degrees C) of the cell preparations just before the incubation was sufficient for making the cells permeable. Localization of enzyme activities was very precise and precipitation of the final reaction product was confined to sites which are known to contain the enzyme under study (granules, mitochondria, lysosomes). These advantages advocate the use of PVA in haematological enzyme cytochemistry and especially for diagnosis of leukemia.
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PMID:Enzyme cytochemistry of unfixed leukocytes and bone marrow cells using polyvinyl alcohol for the diagnosis of leukemia. 280 89

Sodium-potassium adenosinetriphosphatase (Na+-K+-ATPase) is modulated by functional demands. We determine whether Na+-K+-ATPase specific activity was changed by oral administration of different bile salts and whether upregulation in the liver is due to increased numbers of catalytic units. In rats after bile duct drainage for 18 h, Na+-K+-ATPase activity was reduced to 50% of control in liver and ileum but unchanged in jejunum and kidney. Increased Na+-K+-ATPase activity after short-term feeding of bile salts was noted only following trihydroxy bile salts, i.e., taurocholate (100 mg/100 g body wt) increased hepatic Na+-K+-ATPase 143% and ileum 138% above control, whereas jejunum and kidney were unchanged. Chronic feeding of trihydroxy bile salts for 4 days increased hepatic Na+-K+-ATPase (214-260%) and alkaline phosphatase (189-274%), whereas 5'-nucleotidase and Mg2+-ATPase activities were unchanged from control. Plasma membrane Na+-K+-ATPase activity significantly increased as early as 4 h after taurocholate administration, whereas homogenate activity did not rise until 16 h; both reached a new steady state between 24 and 48 h. Sixteen hours after bile salt feeding, increased Na+-K+-ATPase activity was blocked by cycloheximide, and in the liver increased enzyme activity (179%) was associated with a comparable change in sodium-dependent [gamma-32P]ATP binding (162%) to liver plasma membrane fractions. These studies show Na+-K+-ATPase activity adapts selectively in liver and ileum following administration of trihydroxy bile salts, and the process involves increased density of Na+-K+ pump sites on the liver plasma membrane.
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PMID:Selective modulation of hepatic and ileal Na+-K+-ATPase by bile salts in the rat. 283 64

This paper describes in vitro studies on the effects of environmental pollutants (SO2/NOx) in biological systems. Basic physical, chemical and biochemical parameters were analyzed to establish the rate of SO2/NOx absorption by the culture medium. It was shown that the pH remains constant for 24 h of exposure to gas concentrations up to 50 p.p.m. The concentration of ions resulting from absorption of each pollutant in the liquid phase is dependent on their concentration in the gas phase and on exposure time. Short exposure times and high gas dosages resulted in similar doses in the medium as long exposure periods and low gas dosages. The activities of a human serum standard (alkaline phosphatase, ALP; aspartate amino transferase, AST; alanine amino transferase, ALT; gamma-glutamyltransferase, gamma-GT; lactate dehydrogenase, LDH) were determined after gaseous exposure to SO2 and NOx. The results revealed a distinct decrease in the activity of LDH after 1, 3 and 5 h exposure to 200 p.p.m. SO2. The effects of the pollutants were assayed in vitro using fetal hamster lung cells (FHLC), rat hepatocytes and the cell line CO60. For the determination of toxic effects, it was shown that the plating efficiency was a more sensitive parameter than the assay for trypan blue exclusion. Toxicity indicated as an increase of LDH leakage was not observed from FHLC in culture. Instead, a decrease of LDH was found following SO2 exposition. This decrease was similar to that observed for the human serum standard. The induction of DNA single-strand breaks was determined as a measure of genotoxic effects. SO2 application decreased the rate of DNA single-strand breaks induced by N-nitroso-acetoxymethyl-methylamine in both FHLC and in rat hepatocytes. SO2 or NOx treatment of CO60 cells for 1 h did not result in the induction of DNA amplification. HSO3- added directly to the medium as the sodium salt, however, distinctly induced the amplification of SV40 DNA. The amplification rates induced by benzo[a]pyrene or dimethylbenzanthracene were neither influenced by SO2, NOx nor HSO3-. An additive effect of HSO3- with either benzo[a]pyrene or dimethylbenzanthracene for this biological parameter was therefore not observed.
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PMID:Effects of SO2 or NOx on toxic and genotoxic properties of chemical carcinogens. I. In vitro studies. 283 97

Effects of non lethal concentrations of hexavalent chromium on intestinal enzymology of Salmo gairdneri and Dicentrarchus labrax (Pisces). The effects of an exposure to potassium dichromate on intestinal enzyme activities (Alkaline phosphatase, maltase, leucine amino peptidase and ATPases) have been studied on a fresh water fish (Salmo gairdneri) and a salt water fish (Dicentrarchus labrax). Fish were exposed at seasonal temperatures (13 or 21 degrees C) to toxic concentrations equal to 1/10 of the 24 h-LC 50 (i.e. 18 mg/l Cr for trout and 5 mg/l Cr for bass) during respectively 13 and 21 days. Intoxicated trout stopped feeding and showed a decrease in their intestinal weight at the end of the experiments. A decrease of brush border membrane activities (Alkaline phosphatase, maltase and leucine amino peptidase) were also observed. These alterations have been interpreted as the consequence of the chromium induces fasting. Intoxicated bass showed no alterations of their feeding habits. Two specific effects of chromium on enzyme activities have been found: a severe decrease of the alkaline phosphatase activity and an increase of the Na/K ATPase activity. These enzyme activities could be useful indicators of chromium intoxication in marine fish.
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PMID:[Effects of hexavalent chromium at non-lethal concentrations on the enzymology of the intestine of Salmo gairdneri and Dicentrarchus labrax (Pisces)]. 297 85

Activation of the 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] receptor-hormone complex was studied in-vitro using cytosolic preparations of rat osteogenic sarcoma cell ROS 17/2-8 and a DNA-cellulose assay. We found that salt was required for extraction of the unoccupied receptor indicating its possible nuclear localization. The 1,25(OH)2D3 receptor underwent an activation process similar to other steroid hormones which could be stimulated by heat and salt. At physiological ionic strength 100% of the complexes were, however, activated at 2 degrees C, indicating that the activation process is not absolutely temperature dependent. In contrast to other steroid hormones, 30-50% of the complexes were in an activated state in the absence of heat and salt moreover, alkaline phosphatase and ammonium sulphate had no effect on activation. Activation was also stimulated by ATP and ATP plus 8BrcAMP indicating the possible role of phosphorylation in the activation process; however, further work is required to clarify this point.
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PMID:Activation of the 1,25-dihydroxyvitamin D3 receptor in cultured rat osteogenic sarcoma cells. 299 3

Bone density and related biochemical parameters were investigated in institutionalised children and adults with severe handicaps, who were classified according to the degree of limited mobility (group 1, bed-ridden; group 2, capable of crawling; group 3, capable of walking) and according to whether or not they were receiving anticonvulsants. As determined by microdensitometric analysis of radiograms of the second metacarpal bone, bone width (D), bone pattern area (sigma GS) and bone salt density (sigma GS/D) were decreased in the patients, the decreases being most prominent in group 1, followed by groups 2 and 3, in that order. Significant decreases of sigma GS and sigma GS/D, but not of D, were found in patients on anticonvulsant treatment in comparison to patients without therapy. Serum alkaline phosphatase (Al-p) and parathyroid hormone (iPTH) as well as urinary calcium and cyclic adenosine-3',5'-monophosphate (cAMP) excretion were significantly increased in group 1. In comparison to patients without therapy, anticonvulsant-treated children showed significantly decreased levels of serum calcium (Ca), ionised Ca (Ca2+), 25-hydroxy vitamin D3 and urinary phosphate (PO4) excretion, and elevated levels of Al-p, iPTH and calcitonin (iCT). It is suggested that limited physical activity results in a mild hyperparathyroid state, which is aggravated in patients on anticonvulsant treatment.
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PMID:Decreased bone density in severely handicapped children and adults, with reference to the influence of limited mobility and anticonvulsant medication. 300 52

The use of alkaline phosphatase immunocytochemical staining was explored for the rapid diagnosis of poliovirus, adenovirus, herpes simplex virus and cytomegalovirus infections in cell cultures. In this test, viral antigens treated with their relative antibody were incubated with alkaline phosphatase-labelled antisera. The enzyme label was developed with a naphthol salt in the presence of a diazonium salt (Fast Blue) in order to obtain a blue coloured precipitate at the site of the enzyme. It is suggested that this immunocytochemical technique is valuable in the detection of viral infections and would be an appropriate test to use when rapid diagnosis is required.
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PMID:Rapid diagnosis of viral infections by an alkaline phosphatase immunocytochemical method. 300 28


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