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)

Recombinant human granulocyte colony-stimulating factor (rhG-CSF) not only enhanced the growth of HL-60 cells, but also significantly increased NBT-reducing ability and alkaline phosphatase (ALP) activity of the cells, which were enhanced by the treatment with retinoic acid (RA). Protein kinase C inhibitors (H-7 and staurosporine) significantly suppressed this induction of ALP. The pretreatment with RA followed by rhG-CSF treatment showed almost the same degree of ALP activity as that induced by the simultaneous treatment with RA and rhG-CSF. This study suggests that RA and rhG-CSF are the potent inducers of ALP activity of HL-60 cells and protein kinase C is supposed to have a role in this induction of ALP.
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PMID:Alkaline phosphatase activity in the human promyelocytic leukemia cell line, HL-60, induced by retinoic acid and recombinant human granulocyte colony-stimulating factor. 768 28

The breakthrough of chemiluminescence in the field of solution immunoassays and transfer membranes prompted us to explore whether a light-based detection system could provide a gain in sensitivity over chromogenic and FITC markers for nucleic acid and protein detection on histological preparations. A Hamamatsu device and an enhanced chemiluminescence (ECL) luminol substrate of the peroxidase were used to detect epithelial and endothelial components by immunohistochemistry (IHC) and for in situ hybridization (ISH) of papilloma virus DNA. The accuracy of the signal was compared to that obtained with DAB-peroxidase, silver-enhanced DAB-peroxidase, NBT-BCIP-alkaline phosphatase, and FITC. Our results demonstrated the feasibility and high sensitivity of luminescence detection for histological preparations. In part due to the ultrasensitive videocamera and photon-counting imaging, interpretable and reproducible results were obtained within counting times shorter than 5 min, and with dilutions of the primary antibodies 100- to 10,000-fold greater than those used for chromogenic and FITC reactions. As for ISH, with identical concentrations of the HPV 18 DNA probe on HeLa cells, labeling was apparent by luminescence but undetectable with the chromogen. The morphological resolution allowed a discriminatory analysis of the signal. Therefore, at the light microscopic level, enhanced chemiluminescence offers an appealing alternative to FITC and chromogenic markers for detection and quantification of low-concentration molecules.
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PMID:Enhanced chemiluminescence: a high-sensitivity detection system for in situ hybridization and immunohistochemistry. 769 29

In 29 infants with II and III degree of undernutrition, the studies were performed of the granulocyte system including: the number of granulocytes in the bone marrow reserve pool, circulating pool, parietal and total peripheral blood, phagocytosis evaluation, NBT test and the activity of granulocytic enzymes-alkaline phosphatase and peroxidase. The following deviations were found in comparison to children of the same age with normal body weight: 1. Significant decrease of the reserve granulocyte pool in the bone marrow in children with considerable undernutrition, reaching the value of bone marrow reserve in newborn. 2. Increase of phagocytic activity of granulocyte in children with undernutrition, and increased readiness of these cells to intracellular killing. 3. Normal activity of alkaline phosphatase in granulocytes, increased peroxidase activity during the period of considerable undernutrition.
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PMID:[Evaluation of granulocytes in children with malnutrition]. 771 31

Using a digoxygenin-labelled DNA probe derived from the porcine repeat element PRE-1, we have developed a protocol for the detection of transplanted porcine islets and hepatocytes against a background of murine host tissue. Analysis of this probe by Southern blotting indicated that PRE-1 hybridizes to pig genomic DNA but not to human or mouse DNA. On tissue sections, hybridizing probe was detected using alkaline phosphatase-conjugated antidigoxygenin antibody visualized with 5-bromo-4-chloro-3-indolyl-phosphate/4-nitro-blue tetrazolium chloride (BCIP/NBT) substrate. We have demonstrated sensitive and highly specific staining of porcine nuclei in fixed, paraffin embedded tissue sections, and have applied the technique to detect porcine pancreatic islets and hepatocytes transplanted into murine kidney and spleen. Application of this technique include detection of transplanted cells or organs across the variety of xenogeneic barriers.
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PMID:Porcine repeat element DNA: in situ detection of xenotransplanted cells. 777 59

Studied at the cell level of integration were the following indices for nonspecific reactivity: phagocytic activity of leucocytes (PhAL), a parameter characterizing damage to neutrophils (PDN), nitroblue tetrasolium test (NBT-test), the activity of alkaline phosphatase (AlPhA) of neutrophils, and the activity of acid phosphatase (AcPhA) of lymphocytes in EAA (exogenous allergic alveolitis) workers with reference to the service duration. PDN was shown to be raised in EAA workmen who had worked at the integrated poultry farm for 6-10 and 16-20 years. The parameter characterizing damage to neutrophils did not differ from control values in the workmen with 1-5 and 11-15 yr service duration. It was found that the activity of AlPh of neutrophils was decreased in EAA workers with 1-5 and 11-15 yr service duration. The activity of AlPh did not change with 6-10- and 16-20 yr service duration. The results from NBT-test, determinations of PhAL, and the activity of AcPh of lymphocytes showed their elevation in EAA workmen with the service duration 1-20 years. In summary, study at the cell level of integration into the indices of nonspecific reactivity in workmen with 1-2-yr length of service showed a decrease in the activity of AlPh neutrophils and an increase in the activity of AcPh of lymphocytes, PhAL, PDN, NBT-test, suggesting some disturbance in metabolism in leucocytes and involvement of nonspecific mechanisms of protection of organism in EAA.
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PMID:[The cellular level of integration in the body of the workers at a poultry plant with extrinsic allergic alveolitis]. 783 10

Previous evidence has shown that rats with spontaneous hypertension have on average about twice as many circulating leukocytes in comparison with their normotensive counterparts, the Wistar-Kyoto rats. Since such high levels of leukocytes may increase the risk for vascular complications for hypertensive animals, it is useful to ascertain whether a comparable derangement is present in other forms of hypertension. The present study deals with the properties of the circulating leukocytes in rats exhibiting another form of experimental hypertension; Dahl salt-sensitive (Dahl-S) hypertensive rats were compared with Dahl salt-resistant (Dahl-R) control rats. Measurements were performed to determine the following: circulating hematocrit levels, leukocyte counts, differential counts, number of activated leukocytes (by means of nitro blue tetrazolium [NBT] reduction), leukocyte adhesion in vitro and neutrophil CD-18 expression, alkaline phosphatase activity in individual neutrophils and in the plasma, and myeloperoxidase activity in neutrophils. The experimental cohort consisted of Dahl-S and Dahl-R rats maintained for a 6-week period on a 6% NaCl diet. The results show a highly significant elevation in the number of total leukocytes, neutrophil and monocyte counts, and NBT-positive neutrophils and monocytes in Dahl-S but not Dahl-R rats. There was a significant loss of alkaline phosphatase and myeloperoxidase activity in the neutrophils of the salt-treated Dahl-S rats but not in the neutrophils of the untreated Dahl-S or Dahl-R rats. No significant differences were found in neutrophil adhesion under in vitro test conditions between the two strains maintained on the salt diet.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Circulating leukocyte counts, activation, and degranulation in Dahl hypertensive rats. 783 39

In 20 patients with active, endoscopically confirmed duodenal ulcer, serum lysozyme activity, metabolic and phagocytic activity of neutrophils using the NBT spontaneous and stimulated test and stimulated test, and cytoenzymatic reaction for alkaline phosphatase were evaluated before treatment and after two weeks of treatment with ranitidine 150 mg every 12 hours. In some of the studies carried out the differences were obtained between the mean results which turned out to be statistically significant. In the group of patients with duodenal ulcer after two weeks of treatment with ranitidine, a statistically significant increase was found of diluted serum lysozyme activity as compared to the activity of this enzyme before the treatment. After the treatment with ranitidine, a statistically highly significant increase was noted of alkaline phosphatase activity in the neutrophils as compared to the activity of this enzyme assessed in patients with untreated duodenal ulcer. On the basis of the obtained results of studies the conclusions were formulated: 1) The increase of diluted serum lysozyme activity in the patients with duodenal ulcer after two weeks of treatment with ranitidine may evidence the presence of inhibitors of the enzyme in the serum of the studied group. 2) In patients with duodenal ulcer after two weeks of treatment with ranitidine the activity increases of cytoenzymatically determined alkaline phosphatase in the neutrophils, while the cytoenzymatically determined values of the markers in the test with NBT remain unchanged.
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PMID:[Activity of lysozyme in serum and selected granulocytic cytoenzymatic markers evaluated in patients with duodenal ulcer treated with ranitidine]. 797 47

In 21 patients with active, endoscopically confirmed untreated duodenal ulcer and in a control group of 20 persons, the studies were carried out of peripheral blood neutrophils which included: 1) test with the assessment of neutrophil phagocytic activity against live bacterial cells of the Staphylococcus aureus Oxford 209P standard strain and against latex particles, 2) test of nitroblue tetrazolium reduction (NBT test) by neutrophils, 3) evaluation of alkaline phosphatase activity in the neutrophils by a cytoenzymatic technique. In the group of patients as compared with the control group a decrease was observed of neutrophil phagocytic activity against live bacterial cells, and absence of similar differences was noted when latex particles were the stimulating factor. In the studied group an increase of the value of the index of spontaneous NBT reduction by neutrophils was observed as well as a decrease of the value of the index of latex-stimulated reduction of this dye. Besides that, higher alkaline phosphatase activity was observed in the neutrophils in patients as compared to the activity of this enzyme in the neutrophils of healthy persons. It may be supposed that in patients with duodenal ulcer, peripheral blood neutrophils reveal changes in the function of membranous structures of these cells, and disturbances of redox processes evaluated in the test with NBT.
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PMID:[Phagocytosis and NBT test and phosphatase activity of peripheral blood neutrophils in patients with untreated duodenal ulcer]. 797 48

The purpose of this study was to compare, using cell blot analysis, the association of gingival tissue mononuclear cells (GTMC) isolated from lesions displaying histories of early-onset periodontitis (EOP; typically B-lymphocyte dominated) and gingivitis (typically T-lymphocyte dominated) with the B-cell stimulating cytokine, interleukin (IL)-4, and the T-cell stimulating cytokine, IL-2. Eleven EOP patients and 11 age- and gender-similar gingivitis control (GC) subjects participated. Gingival tissue adjacent to the alveolar crest normally removed during surgery was digested in collagenase-containing media and GTMC were isolated by density gradient centrifugation. Cells were separated into four aliquots. One was left unstimulated; the remainder were stimulated for 2 hours with Porphyromonas gingivalis outer membrane protein, mitogen Concanavalin A, or common antigen tetanus toxoid. Cells then were centrifuged onto transfer membranes and incubated in RPMI 1640 media for 6 hours to allow absorption of secreted cytokine. Membranes were treated with monoclonal anti-IL-2 or anti-IL-4, followed by a biotin-conjugated second layer, streptavidin-alkaline phosphatase and nitro blue tetrazolium/5-bromo-4-chloro-indolyl-phosphate (NBT/BCIP) color development. A higher percentage of GTMC from EOP patients were IL-2+ when stimulated with P. gingivalis compared with GTMC from GC patients (20 +/- 2% vs. 12 +/- 2%, P < 0.003). A higher percentage of non-stimulated GTMC from EOP patients produced IL-4 than from GC (22 +/- 4% vs. 6 +/- 3%, P < 0.00007), as well as when stimulated with P. gingivalis (22 +/- 3% vs. 13 +/- 2%, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Gingival cell IL-2 and IL-4 in early-onset periodontitis. 799 15

We describe here a simple method for combining non-radioactive and radioactive in situ hybridization and immunohistochemistry on the same brain tissue section. This approach was first developed on the well-characterized hypothalamo-neurohypophyseal system, facilitating the optimization of the triple-labeling procedure and the verification of labeling specificity. We report the simultaneous detection of vasopressin (VP) mRNA with a digoxigenin-labeled oligonucleotide, oxytocin (OT) mRNA with a 35S-labeled oligonucleotide, and OT peptide in the same 12-microns cryostat section. This was performed on floating sections as follows: first, the two probes were hybridized simultaneously; second, the peptide was detected with an immunoperoxidase-DAB procedure; third, the digoxigenin-labeled probe was detected with an alkaline phosphatase-NBT/BCIP technique; and finally, the 35S-labeled probe was detected by histological autoradiography. We also demonstrate that this approach is suitable for the simultaneous detection of tyrosine hydroxylase and two less abundant mRNAs, vasoactive intestinal peptide and vasopressin mRNAs, in the suprachiasmatic nucleus. The combination of the three techniques did not significantly diminish their specificity or sensitivity. In conclusion, this new method, permitting the simultaneous detection of three different products of gene expression in the same section, could be useful for further analysis of the phenotypic organization and its plasticity in endocrine or neural tissues.
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PMID:Combination of non-radioactive and radioactive in situ hybridization with immunohistochemistry: a new method allowing the simultaneous detection of two mRNAs and one antigen in the same brain tissue section. 809 8


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