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)

Intravenous infusion of zymosan-activated plasma (ZAP) in sheep results in acute lung injury mediated in part by free radical release from stimulated neutrophils that are retained in increased numbers in the pulmonary microcirculation. ZAP infusion is also associated with long-term reduction in elicited superoxide anion generation from segmented neutrophils in the circulating and bone marrow pools for at least 24 h after the infusion. The purpose of our study was to investigate the effect of ZAP infusion on leukocyte counts and neutrophil granule-associated enzyme activity in the circulating and bone marrow pools. Cytochemical methods were used to characterize enzyme activity in primary granules (acid phosphatase and myeloperoxidase) and secondary granules (alkaline phosphatase). During the infusion period, neutrophil differential counts decreased less in venous blood than in matched arterial blood. Release from bone marrow stores was evident as increased numbers of circulating band neutrophils during and after the infusion. Neutrophils in venous and arterial blood smears were degranulated acutely during and 1-3 h after the infusion was stopped. Band and segmented neutrophils in bone marrow also appeared slightly degranulated 1-2 h after the infusion. In vitro experiments showed that band and segmented neutrophils in bone marrow degranulated in response to ZAP incubation. Immature polymorphonuclear leukocytes did not degranulate. Five to 24 h after ZAP infusion, enzyme activity in circulating and bone marrow neutrophils was at baseline levels, suggesting that new cells were being released into the circulating pool because degranulated neutrophils do not synthesize new granules. Another indication of a long-term effect in bone marrow were slight decreases in the percentage of immature polymorphonuclear leukocytes and band neutrophils and a significant decrease in the percentage of eosinophils, all of which coincided with a significant increase in the percentage of segmented neutrophils. Our results demonstrate that circulating complement anaphylatoxins affect neutrophils in bone marrow as well as the vascular space.
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PMID:Infusion of zymosan-activated plasma affects neutrophils in peripheral blood and bone marrow in sheep. 143 61

Four children with the acute leukemia are presented. Their blasts shown the presence of 2 cellular lines markers. Coexistence of markers in the blasts was detected with the technique of double staining the blasts from the bone marrow with: alkaline phosphatase-anti-alkaline phosphatase, and peroxidase with the use of monoclonal antibodies series. Analysis of blasts phenotype with monoclonal antibodies confirm the occurrence of leukemias different from the normally programmed cellular line. Deviations of leukemic cells phenotype may be explained with the fact that leukemogenesis is not an absolute block of cells differentiation but combines maturation disorders and proliferation enabling expression normally absent antigens. It confirms the concept of line preservation and presentation of "earlier frozen" phenotype, and explains the occurrence of leukemias in which blasts present phenotype of one line which does not comply with cell differentiation pattern. Further genotypic studies are necessary to clarify pathogenesis and origin of such blasts. Consequently examination of the larger group of patients with hybrid leukemias will enable conclusions concerning prognostic value of such findings and necessity of introduction of the special therapies.
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PMID:[Hybrid leukemia among acute childhood leukemias]. 143 51

The highest prevalence of testicular cancer occurs in young men with high androgen activity. The presence and distribution of androgen receptors (ARs) was therefore investigated in germ cell neoplasia, using two specific monoclonal antibodies. Tissue samples from 18 patients with seminoma and/or carcinoma-in-situ (CIS) of the testis were examined. An indirect immunohistochemical method with a biotin-streptavidin-peroxidase or an alkaline phosphatase detection system was used. 45% of seminoma samples and 42% of CIS samples were AR-positive with antibody AN 1-15. The values obtained using antibody F 39.4.1 were 44 and 40% respectively. Some differences in specificity between the two antibodies were observed. Unusual granular staining of germ cells in normal testes, also present in malignant germ cells, was noted when antibody F39.4.1 was used. The presence of AR protein immunoreactivity in neoplastic germ cells suggests that androgens may be involved in the pathogenesis of the disease.
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PMID:Immunohistochemical identification of androgen receptors in germ cell neoplasia. 147 24

A direct, double- and triple-staining immunoenzymatic method detected and differentiated sporozoites by color in Anopheles stephensi salivary glands and in mixed sporozoite slide preparations. A double-staining method used beta-galactosidase- and alkaline phosphatase-labeled monoclonal antibodies to the circumsporozoite (CS) proteins of Plasmodium berghei and P. falciparum in mosquito salivary glands. The CS proteins were distinguished clearly by the blue-green and red substrate products of beta-galactosidase and alkaline phosphatase, respectively. A triple-staining method differentiated by color among a mixture of P. falciparum and two strains of P. vivax sporozoites. Monoclonal antibodies to the CS proteins conjugated to beta-galactosidase (P. falciparum), alkaline phosphatase (P. vivax variant), and horseradish peroxidase (P. vivax predominant) readily color differentiated sporozoites by the blue-green, purple-blue, and orange-brown substrate products, respectively. This assay may have potential use in malaria transmission studies, genetic crosses of variant strains of plasmodia to determine assortment of CS antigen alleles, and as a technique to determine the fate of the CS antigen in infected mosquitoes.
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PMID:Immunoenzymatic labeling of multiple plasmodial salivary gland sporozoites in a single test. 155 71

A rapid, sensitive, non-isotopic in situ hybridization histochemistry protocol is presented to study the expression of mRNA at the single cell level in anatomically complex structures of the mammalian central nervous system. The protocol uses digoxigenin-UTP-labeled riboprobes, freefloating sections, and alkaline phosphatase and horseradish peroxidase detection. Modifications have been introduced which preserve the integrity of marker molecules, and as such enable the simultaneous identification and characterization of CNS cell types by tract tracing, histochemical, and immunocytochemical detection of intra- and extracellular markers. All pretreatments that enhance probe penetration have been omitted without substantial loss in sensitivity. The protocol has been successfully extended to vibratome sections with subsequent plastic-embedding and semithin sectioning, which considerably broadens the general applicability of this fast and easy ISHH method.
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PMID:A method of in situ hybridization combined with immunocytochemistry, histochemistry, and tract tracing to characterize the mRNA expressing cell types in heterogeneous neuronal populations. 156 50

Parvalbumin and calbindin, two calcium binding proteins in the nervous system, are present in certain neuronal subpopulations. In the present study a method for a simultaneous demonstration of the both antigens was developed, which labels parvalbumin- and calbindin-containing structures in contrasting colours. A horseradish peroxidase-conjugated second antibody was used for the visualization of the monoclonal anticalbindin antibody, whereas the biotinylated anti-parvalbumin antibody was demonstrated by means of a biotin-streptavidine-alkaline phosphatase system. The method may be useful to classify neuronal populations and to study their morphological relationship.
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PMID:Parvalbumin and calbindin immunoreactivity in the rat brain: a double-immunolabelling method. 158 78

Immunophenotypic analysis of acute leukemias is time consuming and often requires flow cytometric analysis. A 1-hour alkaline phosphatase-labeled streptavidin-biotin immunocytochemical procedure was evaluated as an alternative. Seventeen cases of acute leukemia, including 10 acute lymphocytic (ALL) and 7 acute nonlymphocytic, were phenotyped by the rapid immunocytochemical procedure and the results were compared with standard analyses. In all 17 cases, the diagnoses made using standard cytochemical and immunologic methods were the same as obtained in blinded reviews by rapid immunocytochemical analysis. Nine cases of precursor B-cell ALL were positive for CD19 and/or CD22. Five CD19 + cases of ALL reacted with anti-myeloperoxidase, with one case also positive for CD15. CD15 positivity was confirmed on repeated study as well as with plastic section immunoperoxidase staining. Nine cases of ALL were positive for CD10 and eight were positive for terminal deoxynucleotidyl transferase. One case of ALL marked as T-cell ALL with CD1, CD2, CD3, and CD7. All cases of acute nonlymphocytic leukemia were positive for CD15, CD13, and/or CD33; anti-myeloperoxidase was positive in all but one case of monocytic leukemia. All cases of acute nonlymphocytic leukemia were negative for CD10 and one was positive for terminal deoxynucleotidyl transferase. Acute leukemias apparently may be phenotyped easily and accurately in 1 hour with this immunocytochemical technique, and slides may be stored permanently for review. There was in these 17 cases high correlation of the diagnoses with standard flow cytometric and cytochemical results. This rapid method allows a coordinated evaluation of morphologic features and immunophenotype; the latter features facilitated confirmation of unexpected reactivity of myeloid markers CD15 and MPO-7 in some cases of ALL.
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PMID:Rapid immunocytochemical analysis of acute leukemias. 159 10

Nonradioactive in situ hybridization techniques are becoming increasingly important tools for rapid analysis of the topological organization of DNA and RNA sequences within cells. Prerequisite for further advances with these techniques are multiple labeling and detection systems for different probes. Here we summarize our results with a recently developed labeling and detection system. The DNA probe for in situ hybridization is modified with digoxigenin-labeled deoxyuridine-triphosphate. Digoxigenin is linked to dUTP via an 11-atom linear spacer (Dig-[11]-dUTP). Labeled DNA probes were hybridized in situ to chromosome preparations. The hybridization signal was detected using digoxigenin-specific antibodies covalently coupled to enzyme markers (alkaline phosphatase or peroxidase) or to fluorescent dyes. Color reactions catalyzed by the enzymes resulted in precipitates located on the chromosomes at the site of probe hybridization. This was verified by hybridizing DNA probes of known chromosomal origin. The signals were analyzed by bright field, reflection contrast and fluorescence microscopy. The results indicate that the new technique gives strong signals and can also be used in combination with other systems (e.g., biotin) to detect differently labeled DNA probes on the same metaphase plate.
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PMID:Nonradioactive in situ hybridization with digoxigenin labeled DNA probes. 159 73

In spite of the great variety of enzyme immunoassays (EIA) they can be classified into two groups 'analyte-observed' and 'reagent-observed' assays, depending on their reaction principle. The latter are favored by use of monoclonal antibodies and are characterized by a greater sensitivity, a larger measuring range, a lower susceptibility to disturbing influences. They can be used only for detection of macromolecules. For heterogeneous EIAs to be used on laboratory scale, simple adsorption of antigens and antibodies is still recommendable though affinity constants decrease by at least one order of magnitude and antibody density at the solid phase and analyte binding capacity are not parallel due to increasing steric hindrance. For this reason, the antibody with the higher affinity constant should therefore always be used as solid-phase antibody. Microparticles used as solid phase for heterogeneous assays, due to their very high binding capacity for the analyte and extremely short diffusion distances, guarantee 'one step' assays of only a few minutes. Of the limited number of enzymes suitable as markers in immunoassays, horseradish peroxidase is the enzyme of choice followed by alkaline phosphatase. Although enzyme and enzyme-labelled reagents are detectable by fluorogenic product measuring with a sensitivity, which is 10-1000 times higher than using chromogenic substrates, the sensitivity of the assays can be increased only by factor 2-10. Labelling enzymes cannot only be covalently bound to the antibody, but also via anti-enzyme antibodies. Pros and cons of the different methods of coupling the enzyme/anti-enzyme complex to analyte-containing immune complexes are discussed. Different EIA variants to detect specific antibodies are reviewed. Among them only capture EIAs permit precise isotype analysis of antibodies of a distinct idiotype. Homogeneous EIAs are widely spread for hapten determination but even variants based on proximal linkage are no alternatives to heterogeneous EIAs for determination of macromolecules. Different parameters are defined which permit to assess the quality of an immunoassay and which should be used in routine assays as internal controls in the laboratory.
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PMID:Enzyme immunoassay techniques. An overview. 161 58

We have developed a whole-mount histochemical method to monitor the distribution of expressed genes within the intact, developing vertebrate embryo. Background problems that result from alkaline phosphatase- or horseradish peroxidase-based stains have been minimized, enabling both early and late stages of Xenopus embryogenesis to be monitored. The feasibility and utility of this non-isotopic method has been demonstrated by using a specific DNA probe to localize Xenopus laevis Type II collagen mRNA expression to areas surrounding the vacuoles of the notochord in Stage 30 embryos. Expression expands by Stage 41/42 to form a visually striking distribution pattern that includes a variety of chondrogenic tissues such as the vertebrae, otocysts, mandible, and periocular region. Although these experiments focused on expression of a structural gene, the high resolution and sensitivity of the method should allow it also to monitor expression of less abundant mRNA products of non-structural genes such as transcription factors, cytoplasmic regulators, and growth factors. In addition, this approach should be a successful tool to probe expression in normal and perturbed embryos not only of amphibians but also of other vertebrates, including avians and mammals.
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PMID:Distribution of type II collagen mRNA in Xenopus embryos visualized by whole-mount in situ hybridization. 161 77


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