Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: KEGG:D02003 (NBT)
1,323 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Iron deficiency anaemia was induced in rabbits by repeated bleeding. The leucocyte alkaline phosphatase (LAP) of 26 +/- 28 units was significantly reduced compared with control values of 233 +/- 35 units (P less than 0.001). Leucocyte NBT reduction was also diminished, both in Hanks solution (P less than 0.01) and in autologous serum (P less than 0.001). After administration of iron, these values returned to normal. The results suggest that reduced LAP may reflect a deficiency of iron dependent constituents which are necessary for the integrity of normal granulocyte metabolism.
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PMID:Reduced leucocyte alkaline phosphatase activity and decreased NBT reduction test in induced iron deficiency anaemia in rabbits. 38 63

In 51 untreated cases of ulcerative colitis and Crohn's disease some cellular (neutrophil alkaline phosphatase activity, neutrophil NBT reducing capacity, and neutrophil and plasma lysozyme activities) and humoral (serum orosomucoid and serum haptoglobin) indices of disease activity were quantitated. The most pronounced signs of disease activity, thus, were found in severe cases of ulcerative colitis. Combining lysozyme activities with other disease activity indices seems to facilitate the distinction between severe cases of Crohn's disease and ulcerative colitis. Beyond this the addition of the humoral indices seemed not to offer substantial help.
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PMID:Cellular and humoral indices of disease activity in inflammatory bowel disease. 68 Apr 16

The NBT reduction test and determination of alkaline phosphatase activity in the peripheral blood granulocytes (FAG) were done in 94 subjects including 30 blood donors donating blood for the first time and 64 cases of various haematological syndromes. Raised proportion of formazan granulocytes was found in patients with pancytopenia, acute myeloid leukaemia, chronic myeloid leukaemia during blastic exacerbation, Hodgkin's disease during exacerbation and lymphosarcoma. These results correlated with increased FAG activity. Lower proportions of formazan granulocytes capable of spontaneous reduction of NBT were found in patients with chronic myeloid leukaemia, in immunohaemolytic anaemias and in plasmocytoma. Of all the above syndromes only in chronic myeloid leukaemia impaired ability of formazan cell formation parallelled decreased FAG activity. In the remaining syndromes FAG activity in the granulocytes was normal or raised. In the remissions of Hodgkin's disease a fall was observed in the proportion of formazan granulocytes to values of FAG. In chronic myeloid leukaemia the proportion of formazan cells showed considerable fluctuations and no correlation was observed between the proportion of formazan cells and FAG activity.
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PMID:[Spontaneous nitroblue terazolium reduction test (NBT) by peripheral blood granulocytes in healthy subjects and in some hematologic syndromes]. 105 43

The dose-dependent effect of L-asparaginase (Crasnitin, Bayer) on the serum IgG, IgA and IgM content was studied in 14 children with acute lymphoblastic leukemia. This effect was less evident in the intracellular metabolism of peripheral blood granulocytes (studied by the NBT test), in the myeloperoxidase and alkaline phosphatase activities and in the serum glycogen and lipid content.
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PMID:Immunoglobulin and granulocyte cytochemical reactions in L-asparaginase treated children with acute lymphoblastic leukemia. 107 77

We examined alkaline phosphatase (ALP) activity in the HL-60 cell induced by retinoic acid (RA) and recombinant human granulocyte colony-stimulating factor (rhG-CSF). rhG-CSF induced a small but significant increase of NBT-reducing ability and ALP activity of the HL-60 cells. Among various inducers of cell differentiation, 1,25(OH)2D3 and dimethylsulfoxide (DMSO) caused the increase of the NBT-reducing ability and the suppression of ALP activity induced by rhG-CSF, while RA enhanced both of them. Protein kinase C inhibitors (H-7 and staurosporine) but not a protein kinase A inhibitor (HA1004) significantly suppressed the ALP activity induced by the simultaneous treatment with RA and rhG-CSF.
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PMID:[The effects of retinoic acid and recombinant human granulocyte colony-stimulating factor on alkaline phosphatase activity of HL-60 cells]. 128 12

A new luminescent marker for the immunochemical detection of proteins and nucleic acids on filters is reported. The label consists of inorganic crystals, generally called phosphors, with a particle size of 0.1-0.3 microns, stabilized in suspension with polycarboxylic acids and subsequently conjugated to immunoreactive macromolecules. Immunophosphor conjugates exhibit slowly decaying fluorescence that is strong and practically nonfading and not sensitive to quenching by water molecules. They are therefore suited for conventional fluorescence detection as well as for time-resolved detection. The lifetime of the phosphors was in the micro/milliseconds range upon excitation with ultraviolet light. Proteins or nucleic acids immobilized on nitrocellulose filters were detected immunochemically or by hybridization, using haptenized nucleic acid probes followed by immunochemical detection, respectively. The ultimate detection limit of proteins, using phosphor-labeled macromolecules including an immunochemical amplification step, was found to be 10 fg. The detection limit of nucleic acids was 300 fg for demonstration of hapten-labeled probes and 10 pg in hybridization formats with hapten-labeled probes. The sensitivity of methods using phosphor-labeled macromolecules was in all cases as good as or better than that of methods using alkaline phosphatase developed to NBT/BCIP. The use of immunophosphors for detection of proteins and nucleic acids on Western and Southern blots is demonstrated. Finally, the use of multiple phosphors with different kinetic and spectral characteristics for multiparameter studies is indicated.
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PMID:Immunochemical detection of proteins and nucleic acids on filters using small luminescent inorganic crystals as markers. 141 29

We produced monoclonal antibodies against the major allergen of Japanese cedar (Cryptomeria japonica) pollen, Cry j I. KW-S10 antibody reacted only with Japanese cedar pollen and KW-S91 antibody reacted to most angiospermae pollens as well as Japanese cedar pollen. Using these antibodies, we devised a new counting method of Japanese cedar pollen allergen particles by an immunoblotting technique. Airborne pollen allergens were collected on vaseline coated glass slides or Burkard's sampling tape and were transferred onto polyvinylidene difluoride (PVDF) membranes. The membranes were treated with anti-Cry j I monoclonal antibody conjugated with alkaline phosphatase. Pollen allergens were detected as spots on the membranes after staining with phosphatase substrate (BCIP/NBT). This method using KW-S10 antibody measured only the amounts of allergen from Japanese cedar pollen, while with KW-S91 antibody, the method measured the amounts of pollens which have antigenicity in common with Japanese cedar pollen.
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PMID:[A new counting method of airborne Japanese cedar (Cryptomeria japonica) pollen allergens by immunoblotting using anti-Cry j I monoclonal antibody]. 144 17

To avoid extensive manipulation for the purification of RNA from cells, several methods were evaluated for the direct release of RNA from influenza virus infected cells and supernatants using slot blot hybridization and non-radioactive probes. Treatment with an equal volume of 10 M aqueous guanidine hydrochloride produced the best hybridization signal. Less, but significant amounts of RNA were also released using the following treatments: dilute alkali (final concentration of 0.16 M NaOH) or 100 degrees C/5 min or RNA sample buffer containing formamide/formaldehyde, then heating at 65 degrees C/10 min. Despite the presence of large amounts of cell debris, RNA from guanidine hydrochloride treated whole cell extracts bound quantitatively to the positively charged nylon membranes. The sensitivity of RNA detection when whole cell extracts treated with guanidine hydrochloride were probed with a digoxigenin labelled cDNA probe was similar to the detection of RNA in highly purified, protein free samples. Three positively charged membranes were tested (from Amersham, ICN and Boehringer Mannheim) using two alkaline phosphatase substrates, NBT-X phos, and a chemiluminescent substrate, 3-(2'-spiroadamantane)-4-methoxy-4-(3'-phosphoyloxy)-phenyl- 1-1,2-dioextane (AMPPD) and a peroxidase substrate, tetramethylbenzidine (TMB). The Boehringer Mannheim membrane had the highest sensitivity for the alkaline phosphatase substrates, but the peroxidase reaction with the TMB substrate was the most consistently sensitive, irrespective of which membrane was used. The ability to quantitatively detect RNA from whole cells without any purification will allow the rapid screening of large numbers of samples for specific RNA species in research or diagnostic laboratories.
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PMID:Rapid treatment of whole cells and RNA viruses for analysis of RNA by slot blot hybridization. 162 16

A rapid method for qualitative and quantitative detection of specific oral microorganisms from subgingival dental plaque is described. Plaque samples were suspended in phosphate-buffered saline containing protease inhibitors and 0.5% formaldehyde, briefly sonicated to disperse bacterial aggregates, and applied to nitrocellulose membranes in a slot blot manifold. Subsequent incubations with species-specific rabbit antibody and anti-rabbit antibody-alkaline phosphatase conjugate and development with BCIP-NBT substrate resulted in an easily discernible, permanent stain being deposited at the sample application site. Comparison with known concentrations of pure, cultured microorganisms applied to the same membranes permitted qualitative or semiquantitative plaque characterization by visual inspection. Analysis of the blots with a computer-linked flatbed scanner provided quantitative data on microbial content. The reproducibility of the results (standard error of the mean, less than 10%) obtained with slot immunoblotting greatly exceeded that of the results obtained with immunofluorescence analysis (standard error of the mean, greater than 57%). Because it is versatile, rapid, sensitive, reproducible, permanent, and relatively inexpensive, slot immunoblotting lends itself to use in large-scale investigations for the detection and quantitation of specific microbial species.
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PMID:Slot immunoblot assay for detection and quantitation of periodontal disease-associated microorganisms in dental plaque. 166 11

We describe a new, simple, rapid, and sensitive nonradioactive technique for the analysis of genetic variations. Genomic DNA was amplified using polymerase chain reaction and amplified DNA was hybridized, with digoxigenin (DIG)-labeled sequence-specific oligonucleotides. High specificity and sensitivity was achieved when labeling the sequence-specific oligonucleotide at the 3' end with only one DIG using digoxigenin-11-2',3'-dideoxy-uridine-5'-triphosphate and DNA deoxynucleotidylexotransferase. The hybridized probes were detected using antidigoxigenin alkaline phosphatase, fab fragments, and X-phosphate/NBT for visualization. This method was applied to the analysis of HLA-DR4-DRB1 alleles in polymerase chain reaction-amplified genomic DNA and resulted in highly specific and sensitive hybridization signals discriminating even in cases of a one-base-pair mismatch. This technique is particularly suited for HLA oligotyping because it allows the use of tetramethylammonium chloride for the simplification of hybridization and washing conditions.
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PMID:Nonradioactive HLA class II typing using polymerase chain reaction and digoxigenin-11-2'-3'-dideoxy-uridinetriphosphate-labeled oligonucleotide probes. 167 54


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