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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)
An immunobinding assay capable of distinguishing between Mycoplasma felis and Mycoplasma gateae was developed. Nitrocellulose was used as the solid support. Polyclonal rabbit antiserum against M. felis or M. gateae was used in the assay. Binding of the specific rabbit antiserum was detected by
alkaline phosphatase
-conjugated goat anti-rabbit immunoglobulin and an
alkaline phosphatase
substrate (
Nitro Blue Tetrazolium
-5-bromo-4-chloro-3-indolyl phosphate). The immunobinding assay was effective in the detection of feline mycoplasmas on agar plates, on primary isolation plates, in broth cultures, and in mixed cultures. No cross-reactions were observed with other related mycoplasmal species. The assay was cheap to perform and easy to interpret, and it required little technical time.
...
PMID:Identification of Mycoplasma felis and Mycoplasma gateae by an immunobinding assay. 239 8
Recent findings that retinoic acid (RA) induces terminal granulocytic differentiation of the human promyelocytic leukemia cell line HL-60 in vitro and blast cell maturation in patients suffering from acute non-lymphocytic leukemia (ANLL) prompted an investigation on the ability of this agent to induce terminal maturation in blast cells from ANLL patients in vitro. We tested the ability of RA at 3 x 10(-6) M, 3 x 10(-7) M and 3 x 10(8-) M concentrations to induce differentiation in blastoid cells from 16 patients with ANLL using cytochemical and cytologic parameters, in addition to cytofluorometric methods. Leukemic cells in primary culture from all the patients underwent cytochemical and biochemical changes after treatment with RA. However, the extent of differentiation-positive cell clones (D+ clones) varied from patient to patient. Morphologic maturation was observed in a significant number of bone marrow samples. Leukocyte
alkaline phosphatase
and
NBT
reduction ability of cells, which are biochemical markers of granulocytic differentiation, were also significantly increased with a simultaneous decrease in DNA and RNA synthesis (which was estimated using a Phywe ICP-11 impulse flow cytometer).
...
PMID:Multiparametric evaluation of retinoic acid-induced terminal differentiation of blastoid cells from acute non-lymphocytic leukemia patients in vitro. 248 50
Cytochemistries were made in 55 patients with meningitides of different etiology to study the functional metabolic activity of neutrophil leukocytes. At the same time consistent and different changes were discovered in the content of glycogen and cationic protein, in the activity of acid and
alkaline phosphatase
and myeloperoxidase as well as in the indicators of the
NBT
test, depending on the etiology, stage, gravity and the character of complications. In patients afflicted with meningitides, different degree of the activity of intracellular components and of the indicators of the
NBT
test characterizes nonspecific responsiveness and is of differential-diagnostic importance.
...
PMID:[Intracellular metabolism and leukocyte functional activity in patients with meningitis of various etiologies]. 263 83
We described rapid methods to detect Y-specific repeated DNA sequences in cytological preparations using in situ hybridization. A human Y chromosome specific DNA probe with an insert equivalent to that in pHY2.1 was labelled with [alpha-32P]dCTP or photobiotin, and hybridized to chromosome preparations. Signals were visualized specifically on Y chromosomes after 1 day's autoradiography or a couple of hours treatment with streptavidin
alkaline phosphatase
/BCIP/
NBT
. These methods are useful for molecular confirmation of Y-autosomal translocations.
...
PMID:Rapid methods to visualize Y-specific repeated DNA sequences in cytological preparations. 265 88
We compared the sensitivity of a chemiluminescent substrate 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy)phenyl- 1,2-dioxetane (AMPPD) and a chromogenic substrate 5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium (BCIP/
NBT
) for detection of an
alkaline phosphatase
label in a hepatitis B virus "core antigen" DNA (HBVc) probe hybridization assay. Chemiluminescent signal obtained from AMPPD hydrolysis by
alkaline phosphatase
was detected with Polaroid Instant Black and White Type 612 film. The chemiluminescent assay detected 1.18 x 10(6) copies of HBVc plasmid DNA in 30 min. By comparison, 9.8 x 10(7) copies of DNA could be measured using chromogenic BCIP/
NBT
substrate within the same incubation time. After further development, the chemiluminescent endpoint permitted detection of 4.39 x 10(4) copies of HBVc plasmid DNA in 2 h.
...
PMID:A comparison of chemiluminescent and colorimetric substrates in a hepatitis B virus DNA hybridization assay. 281 49
A number of immunocytochemical detection systems for determining the chromosomal localization of specific nucleic acid sequences by non-radioactive in situ hybridization have been compared. The procedures were: 1. the peroxidase/diaminobenzidine (PO/DAB) combination, either or not gold/silver intensificated; 2.
alkaline phosphatase
marking using the nitro-blue tetrazolium plus bromochloro-indolyl phosphate substrate combination (AP/
NBT
+ BCIP); and 3. immunogold with or without silver enhancement. The procedures were first tested and optimized in dot blot experiments and then applied to in situ hybridization. As hybridization probes, both a middle-repetitive and a unique sequence (modified with 2-acetylaminofluorene (AAF] were used. The advantages and disadvantages of the various methods for reflection contrast (RC) or transmission electron microscopic (TEM) visualization of hybrids are discussed.
...
PMID:Non-radioactive in situ hybridization. A comparison of several immunocytochemical detection systems using reflection-contrast and electron microscopy. 361 Jun 73
The leukocytes of patients with chronic granulomatous disease (CGD) may be identified by their failure to reduce
Nitro Blue Tetrazolium
(
NBT
) during phagocytosis. This reaction, normally detected in the phagocytic vacuole, is absent or delayed in CGD monocytes and eosinophils as well as in neutrophils, even though sonicates of normal and CGD leukocytes contain equal activities of a cyanide insensitive enzyme system capable of reduction of
NBT
in the presence of pyridine nucleotide. Enlargement of CGD phagocytic vacuoles appears to be inhibited. Histochemical estimates of the rate of release of
alkaline phosphatase
are normal in CGD cells. Peroxidase activity is released from CGD cells, but the rate appears to be somewhat slower than normal in some cases. The latter observation may be explained by the increased intensity of the peroxidase stain in resting and phagocytizing CGD cells. The severity of the defect in
NBT
reduction within the phagocytic vacuoles of the leukocytes of patients and carriers is more variable than was previously appreciated. Some female carriers have profoundly reduced dye reduction and others are nearly indistinguishable from normal. Three brothers with CGD demonstrated significant, albeit delayed,
NBT
reduction in phagocytic vacuoles during prolonged incubation of their leukocytes. No obvious relationship exists, however, between the rate of reduction of
NBT
in vacuoles and the clinical severity of the disease.
...
PMID:Failure of nitro blue tetrazolium reduction in the phagocytic vacuoles of leukocytes in chronic granulomatous disease. 538 30
In 106 workers (47 women and 59 men) being in professional contact with organic solvents containing benzene and its homologues during 1 to 122 months the cytochemical examination of peripheral blood neutrophils has been performed. The patterns of neutrophil functional activation have been noted expressed in increased activities of acid phosphatase and beta-glucuronidase, increased
NBT
reduction and diminished glycogen reserves. Those changes were accompanied by diminished peroxidase and
alkaline phosphatase
activities. The stimulated
NBT
reduction, elevated in majority of workers, exhibited negative correlation with the exposure time what indicates the practical value of that test monitoring the biological effects of professional contact with the solvents.
...
PMID:Cytoenzymatic studies on neutrophils in workers having contact with organic solvents containing benzene, toluene and xylene. 616 42
The group of aged subjects being 66 to 97 years old was compared with the middle-age group with regard to various immunological and cytochemical indices related to lymphocytes and neutrophils. The aged showed a lowered count and percentage of T cells, increased count and percentage of "non-B, non-T" lymphocytes, increased percentage of B cells. These alterations in the composition of lymphocyte subpopulation were associated with characteristic patterns of damage affecting the enzyme-positive lysosomal apparatus of lymphocytes with regard to acid phosphatase, beta-glucuronidase and N-acetyl-beta-D-glucosaminidase. There was a hundredfold smaller number of cells having intact enzyme-positive lysosomes in the aged than in the group of comparison. The changes mentioned above were also associated with the intracellular accumulation of glycogen in lymphocytes, decreased concentration of IgG and IgM in the serum and various changes in IgA concentration. Neutrophils of the aged were fewer in the blood of the aged than in younger subjects. However, an increased activity of myeloperoxidase,
alkaline phosphatase
, N-acetyl-beta-D-glucosaminidase, and an increased content of glycogen and lipids could be found in these cells.
NBT
-positive neutrophil numbers in the aged were lowered if the stimulated test was used and if there were no changes of the spontaneous test.
...
PMID:Immunological and cytochemical indices of white blood cells in old age. 619 50
A new method is described which is suitable for reliably analysing apoptotic fragmentation in small amounts of DNA. After isolation, DNA was labelled with biotin-4-dUTP using Klenow polymerase. Then DNA was size-separated by agarose gel electrophoresis, blot transferred and subsequently visualized by the streptavidin
alkaline phosphatase
-BCIP/
NBT
procedure. This non-radioactive method was used to detect apoptotic DNA in rat pheochromocytoma PC12 cells, treated with tributyltin (1 nM). While only 30 ng of DNA is required for analysis of apoptotic DNA using the new blot technique, 100-fold more material is needed to identify the fragmentation of DNA after separation by agarose gel electrophoresis and direct staining with ethidium bromide. In a further set of experiments, rat cortical cells were incubated with human immunodeficiency virus type 1 viral glycoprotein of M(r) of 120 kDa (gp120) to induce apoptosis. More than 0.3 ng of gp 120/ml are required to detect apoptotic DNA by the direct procedure; only 0.1 ng gp120/ml or less were sufficient to document clear DNA fragmentation using the non-radioactive blotting technique described here. These results demonstrate that the new procedure can be used to analyse very small amounts of apoptotic DNA and shows that gp120-induced apoptosis can be measured at low concentrations of the viral protein.
...
PMID:A non-radioactive, sensitive method for the detection of DNA fragmentation in apoptotic cells (rat pheochromocytoma PC12 and rat cortical cells). 752 53
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