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)

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

Lysed granule fraction was prepared from splenic adherent cells from 0-day-old to 2-month-old chickens and was examined antibacterial activity to Salmonella pullorum and Salmonella sentfenberg. The activity of lysed granule fraction from 2-month-old chickens was exhibited at pH 5.0 but not exhibited at pH 6.0 or 7.0 to both serotypes of Salmonella. At pH 5.0, lysed granule fraction from 0-day-old chickens revealed little antibacterial activity to S. pullorum but the activity increased with increasing age. At 240 min of incubation, 60% and 95% of S. pullorum were inactivated in lysed granule fractions from 7-day-old and 2-month-old chickens, respectively. High antibacterial activity to S. senftenberg was observed in lysed granule fraction from 2-month-old chickens in which 99.6% of the bacteria was inactivated during 240 min; In lysed granule fractions from 0-day old and 7-day-old chickens, 90% of S. senftenberg was inactivated during 60 min but the number of bacteria increased conversely after incubation for 240 min. Changes in enzymatic activities of 4 enzymes in the lysed granule fractions from various ages of chickens were examined. beta-Glucronidase increased statistically significantly to 2.4 and 7.6 times from 0-day-old to 7-day-old and 2-month-old chickens, respectively. Acid phosphatase and alkaline phosphatase increased slightly but not significantly different, and enzymatic activity of myeloperoxidase was scarcely detected.
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PMID:Age-dependent resistance of chicken of Salmonella in vitro: antibacterial activity of lysed granule fraction of splenic adherent cells. 1 89

Isopycnic sedimentation has been used to separate granulocytes of varying stages of maturity from the bone marrows of normal rabbits and rabbits stimulated to undergo an intense inflammatory response. The separated cell populations were in turn utilized to study the specific activities of six intracellular enzymes. The study revealed an increase with cell maturation in the specific activities of myeloperoxidase, NADPH oxidase, alkaline phosphatase and acid phosphatase in normal animals; in stimulated animals only myeloperoxidase and NADPH oxidase increased significantly with cell maturation. Glucose-6-phosphate dehydrogenase showed no change in specific activity in all animals studied. Malate dehydrogenase tended to show a specific activity decrease in the maturing cells of normal but not in those of stimulated animals.
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PMID:Characterization of marrow granulocyte development: enzyme-specific activity profiles in response to inflammatory reactions. 2 66

A transplantable myelogenous leukemia of an inbred Wistar/Furth rat has been established in tissue culture and cloned. The resulting transplantable leukemia line demonstrates in vitro doubling time of 20 hr, colony-forming efficiency of 5% in liquid and methylcellulos-containing medium, and a saturation density of 3.0 x 106 cells/sq cm in liquid medium. Following intraperitoneal inoculation, newborn rats developed solid tumors, ascities, and leukemia with ld50 of5 x 103 cells and mean latency of 60 days. The tumor cell morphology was consistent with that of acute myelogenous leukemia. Histochemical staining for myeloid enzymes revealed no evidence of myeloperoxidase, esterase, or leukocyte alkaline phosphatase; however, fluorescent antibody staining for lysozyme was markedly positive. Serum, urine, and ascitic fluid from rats with transplanted leukemia also contained elevated levels of lysozyme. There was no detectable type-CRNA virus production by this cell line after as long as 100 days in vitro. This inbred rat myelogenous leukemia should provide a useful model for studies of chemotherapy and immunoltherapy of human acute myelogenous leukemia.
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PMID:Acute myelogenous leukemia of the Wistar/Furth rat: establishment of a continuous tissue culture line producing lysozyme in vitro and in vivo. 4 87

The haematopoietic tissue in the supraneural organ of the freshwater river lamprey (Lampetra fluviatilis L. Gray) was studied in sexually immature animals. Besides erythro- and granulopoietic elements, macrophages, reticular cells, fibroblasts and glycogen-rich fat cells were seen. Developing granulocytes of the lamprey contain one type of azurophil granules originating from small cytoplasmic (Golgi) vesicles. The lamprey's azurophil granulocytes seem to be homologous with those of fishes. However, the granulocytes of fishes, studied thus far, show granules with only one type of inclusion, whereas in lamprey the granulocyte inclusions are variable in size and shape. Thus, lamprey granulocytes are, in this respect, reminiscent of similar cells of higher vertebrates. The PAS and alkaline phosphatase reactions, common markers of vertebrate neutrophil leucocytes, are very weak in the haematopoietic tissue granulocytes of the lamprey, and intense in the blood cells of the same animal. Lamprey granulocytes, similarly to the granulocytes of Chondrostei and Elasmobranchiata, do not stain with peroxidase, naphthol-AS-D-chloroacetate esterase and sudan black B. The haematopoietic tissue contains a relatively high number of degenerated granulocytes.
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PMID:The haematopoietic supraneural organ of adult, sexually immature river lampreys (Lampetra fluviatilis [L.] Gray) with particular reference to azurophil leucocytes. 6 2

Four cytochemical reactions widely used for hematological diagnosis (Myeloperoxydase, leukocyte alkaline phosphatase, PAS and Sudan Black) were carried out in 46 infants admitted into a neonatology unit (14 preterm babies, 19 small for date newborn infants (SDB) and 13 preterm with retarded intra-uterine growth). The MPO, polymorphonuclear PAS and Sudan Black levels are comparable in the three groups of infants and the same as in normal full-term newborn babies. On the other hand, the LAP level, identical in preterms and SDB, is lower than in full-term infants but higher than in adults. Lymphocyte PAS value increases progressively with age. Better interpretation of results would be gained with knowledge of normal ranges for these four parameters in children. The authors suggest that these reactions should be used in the field of general metabolic diseases, susceptibility to infection and genetic diseases with or without chromosal abnomaly.
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PMID:Leukocyte cytochemical reactions in preterm and small for date babies. 6 99

Eosinophilic leukocytes may accompany a great variety of disorders and different types of acute leukemias. The most striking morphologic feature of eosinophils is their specific granules, but morphology alone often is insufficient to differentiate normal from abnormal eosinophils. Cytochemically, the eosinophils were considered "normal" when they did not contain alkaline phosphatase, chloroacetate esterase, toluidine blue metachromasia, Astra blue positivity, and specific PAS-positive granules, but did have peroxidase and cyanide-resistant peroxidase activities, Sudan black positivity and moderate naphthol-AS esterase or alpha-naphthyl esterase and acid phosphatase positivities. In seven cases of acute leukemias (two acute myeloblastic and five myelomonocytic), in contrast with their normal behaviour, the eosinophils show "abnormal" cytochemical positivities consisting of chloroesterase activity, PAS and Astra blue positivities of the specific granules, toluidine blue metachromasia, and cyanide-resistant peroxidase of a few specific granules. Cytochemical investigations may provide additional criteria for evaluating the abnormality of the eosinophilic cell in leukemias.
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PMID:Cytochemical "normal" and "abnormal" eosinophils in acute leukemias. 7 Jan 68

The use of alkaline phosphatase in an immunoenzymatic procedure for the localisation of antigens in paraffin sections or cell smears is described. The results of this method, when applied to the detection of immunoglobulins, lysozyme, or lactoferrin, were comparable in intensity and clarity to those obtained with the PAP immunoperoxidase procedure. Furthermore, double immunoenzymatic labelling (with alkaline phosphatase and peroxidase) of two cellular constituents in a tissue section is possible, the brown peroxidase reaction product contrasting well with the blue alkaline phosphatase product. Since the two antibody 'sandwiches' are applied simultaneously rather than sequentially the total duration of this double immunostaining procedure is only a few minutes longer than that required for detection of a single antigen. It was also found that the unlabelled antibody immunohistological procedure (whether used in conjunction with alkaline phosphatase or peroxidase) can be shortened without loss of sensitivity by carrying out two of the incubation steps simultaneously.
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PMID:Alkaline phosphatase and peroxidase for double immunoenzymatic labelling of cellular constituents. 7 79

The aim of the present investigation was to test a procedure useful for the electron-microscopic in situ identification of the presumptive cariogenic microorganism Streptococcus mutans (serotype d) growing in the human dental plaque. For this purpose, different parameters of an indirect immunohistochemical method were tested in three sets of experiments. In experiment set I, all serological and histochemical procedures were performed en bloc on specimens fixed only with glutaraldehyde before embedding in Vestopal W. Different marking substances, such as ferritin, alkaline phosphatase and horseradish peroxidase (HRP) were tested. The en bloc method using HRP-labelled antibodies was found useful for staining of in vitro-grown bacteria but failed when applied to dental plaque. In experiment set II, ultrathin sections of glutaraldehyde-fixed and glycol methacrylate-embedded in vitro-grown bacteria were section-stained. The bacteria became outlined with a highly electron-dense HRP reaction product which accumulated on top of the cell envelope. The same type of HRP reaction product was found on some bacteria in the 2-day-old dental plaque after section-staining. This system was further tested in a series of controls also using consecutive serial sections. In exp. set III, a number of different stationary and transient oral bacteria were immunohistochemically section-stained. A cross-reaction with bacteria belonging to S.salivarius was discovered and removed by absorbing the anti-S.mutans serotype d serum with S.salivarius (NCTC 8618).
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PMID:Some immunohistochemical experiments aiming at the electron-microscopic in situ identification of a dental plaque microorganism--Streptococcus mutans. 8 15

It was found that neutrophils in untreated uraemic patients as well as in subjects on regular dialysis treatment displayed higher activity of acid phosphatase, alkaline phosphatase and peroxidase. Spontaneous reduction of nitro blue tetrazolium (NBT) by granulocytes was also higher in both groups in comparison to controls. Stimulation with latex particles gave similar results of NBT reduction in investigated patients and controls. Lymphocytes also showed an increase in acid phosphatase activity if compared to healthy persons. It seems possible that granulocytes which take part in unspecific defense mechanisms are more active in uraemic patients due perhaps to subclinical infections.
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PMID:The enzymatic activities and NBT reduction test of granulocytes in untreated and dialysed uraemic patients. 10 90


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