Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The objective of the present study was to characterize the interaction between human platelets and vaccinia virus and to examine possible impairment of platelet functions. The vaccinia virus was selected for our model system because it lacks detectable neuraminidase activity. Platelets were incubated with purified viral particles labeled with 3H-thymidine and binding parameters were analyzed. Binding reached saturation with an average of 5 particles/platelet. It was not affected by the plasma but was sensitive to temperature and to metabolic inhibitors. 3H-thymidine-labeled vaccinia virus and formaldehyde-fixed platelets were used to measure viral adsorption. The adsorption was temperature-independent but was affected by ionic strength, indicating electrostatic interactions. Treatment of the fixed platelets with neuraminidase or with alkaline phosphatase reduced viral adsorption, indicating that sialate and phosphate residues on the platelet surface may be involved in the adsorption. Platelet activities were markedly affected by vaccinia virus. The virus caused a dramatic 14C-serotonin release with no added inducer. The release was inhibited by aspirin, a known inhibitor of serotonin release related to prostaglandin synthesis. Furthermore, the virus inhibited platelet aggregation, induced by either ADP, collagen, or thrombin. This study demonstrates that although vaccinia virus lacks neuraminidase activity, it does bind to platelets and affects their function.
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PMID:Interaction between vaccinia virus and human blood platelets. 705 66

Segmental and complete hepatic artery embolization with Ivalon (polyvinyl alcohol) particles (0.25 to 1 mm) was performed in 12 dogs to evaluate hepatic function alterations and histopathological changes. In dogs undergoing segmental embolization, liver function alterations were minimal and the liver was normal, both grossly and microscopically, at autopsy. In dogs undergoing complete hepatic embolization, only two had significant elevation of SGOT and alkaline phosphatase levels, which normalized in 2 and 4 weeks; one of these two dogs also had an elevated bilirubin level. A focal hepatic infarct was observed both grossly and microscopically in one dog and only microscopically in four dogs. Two dogs died of pancreatic abscess due to unintentional pancreatic embolization. The study suggested that segmental embolization with Ivalon particles was well tolerated by dogs, and complete hepatic embolization resulted in hepatic function changes and focal infarction comparable with Gelfoam embolization.
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PMID:Experimental canine hepatic artery embolization with polyvinyl alcohol foam particles. 712 82

In situ hybridization (ISH) of somatostatin (SOM) mRNA was carried out on sections of rat brain using an alkaline phosphatase (AP) coupled oligonucleotide probe. Different hybridization and AP development conditions were tested for qualitative and quantitative detection of target mRNA on sections of unfixed tissue. Hybridization signal intensities after 24 h of hybridization were high. Comparison with adjacent formaldehyde-fixed tissue sections and hybridization for various lengths of time (2-42 h) indicated that in unfixed tissue retention of SOM mRNA was at least as high as after fixation, and that the mRNA was not degraded during hybridization. The use of tetranitroblue instead of nitroblue tetrazolium chloride in the AP detection medium provided a superior signal-to-noise ratio, and medium stability was improved for quantitative studies on unfixed sections by adding 10% polyvinyl alcohol at pH 8.5. Microphotometric measurements of mean optical densities (MOD) of the formazan reaction product in a defined area within individual neurons of the lateral central amygdaloid nucleus showed a linear increase over the first 23 h of AP reaction time. The mean MOD values per neuron were comparably high in various equally thick sections of the nucleus and increased with section thickness in a linear manner. The findings indicate that the ISH and detection reagents penetrate the entire section and that there is a linear relationship between the amount of AP reaction product measured and the amount of mRNA present in the measured area. Thus, ISH using an AP-coupled oligonucleotide on sections of unfixed tissue appears suitable for quantitative mRNA detection.
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PMID:Qualitative and quantitative detection of alkaline phosphatase coupled to an oligonucleotide probe for somatostatin mRNA after in situ hybridization using unfixed rat brain tissue. 758 53

A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection. Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.
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PMID:Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19. 763 29

NADPH diaphorase is a histochemical activity which, in formaldehyde-fixed tissue, is rather specific for nitric oxide synthase. Recently, it was shown that NADPH diaphorase activity is inhibited by ethylenediaminetetraacetic acid (EDTA) in neurons but not in the choroid plexus epithelium. The present study, while confirming these results, demonstrates that the apparent sensitivity of NADPH diaphorase for EDTA reflects only the dependence of malic enzyme, which is used as the source of reduced cofactor, on Mg2+ or Mn2+ ions. Furthermore, evidence is provided that the apparent EDTA-insensitive NADPH diaphorase activity in the choroid plexus reflects the activity of alkaline phosphatase in conjunction with NADH diaphorase. Apart from these pitfalls, the use of the indirect, malic enzyme based method for NADPH diaphorase was found to cause much higher background staining compared to the direct method using NADPH, and is therefore proposed to be abandoned.
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PMID:NADPH diaphorase is not inhibited by ethylenediaminetetraacetic acid and is not specific for nitric oxide synthase in the choroid plexus of rat and mouse. 773 45

The sensitivity and specificity of 3 avidin-biotin complex (ABC) immunostaining systems were compared on paraffin-embedded tissues from African swine fever virus (ASFV)-infected pigs. Results were also compared with immunofluorescent detection on cryosections of the same tissue for optimal detection of ASFV antigen. The ABC-alkaline phosphatase (ABC-AP) and ABC-peroxidase (ABC-PO) systems were at least as sensitive as direct fluorescent antibody (FA) and 10-fold more sensitive than the ABC-glucose oxidase system. Three ABC-AP and 2 ABC-PO chromagens with different counterstains were compared. In addition, 2 fixatives, 2 biotinylation procedures, 7 endogenous peroxidase blocking regimes, 6 tissue adhesives, and 3 mounting media were compared. The ABC-AP system with a red chromagen and hematoxylin counterstaining was preferred and most closely approximated routinely stained pathologic sections. Fixation in paraformalde-hydelysine-periodate fixative preserved ASFV antigen for research studies for at least 3 years. Formalin-fixed tissues retained some staining for up to 10 years.
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PMID:A comparison of three avidin-biotin complex immunoenzyme systems for detection of African swine fever virus antigen in paraffin-embedded tissues. 777 59

Cytospins of a human breast cancer cell line (MCF-7) were studied for the expression of PCNA, a cell cycle-related protein, using a variety of fixation and immunostaining procedures. The best fixative for PCNA was found to be buffered formaldehyde solution at 4 degrees C followed by methanol at 20 degrees C, whereas alcoholic fixatives decreased greatly the PCNA immunoreactivity. Air-drying procedures of cytospins prior to and after fixation greatly undermined the PCNA immunostaining. A modified immunoperoxidase method provided a stronger staining of the PCNA-reactive cells than the alkaline phosphatase anti-alkaline phosphatase (APAAP) technique. PCNA immunoreactivity could be maintained up to 2 mo, putting slides in methanol at -20 degrees C. In conclusion, our report indicates that PCNA is a labile antigen, which may critically be affected by temperature and air-drying procedures.
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PMID:Methodological aspects of the immunostaining of proliferating cell nuclear antigen (PCNA) in cytospin preparations of MCF-7 cell line. 791 57

We tested several modifications of paraffin embedding technique when examining bone marrow biopsies in order to achieve an optimal combination of morphology and antigen preservation. Short pre-fixation in formalin followed by mixture of formalin and 10% formic acid were found optimal for quick tissue processing where electron microscopy was not planned. Formalin pre-fixation prevents the occurrence of imperfect stainability of superficial parts of trephine cylinders caused by an effect of formic acid on non-fixed bone marrow tissues. Decalcification in mixture of formol and EDTA is recommended if there is time enough. The supersensitive streptavidin-biotin detection system with alkaline phosphatase was found to provide the best results.
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PMID:[Fixation and decalcification in trephination biopsy]. 792 18

We describe a fast light microscopic procedure for the simultaneous enzyme cytochemical detection of three different DNA target sequences in contrasting colors in both interphase and metaphase cell preparations. Chromosome-specific DNA probes labeled with either biotin, digoxygenin, or fluorescein were hybridized as a mixture and detected clearly and accurately by precipitates of the horseradish peroxidase-diaminobenzidine (PO-DAB, brown color), alkaline phosphatase-Fast Red (APase-Fast Red, red color), or horseradish peroxidase-tetramethylbenzidine (PO-TMB, green color) reaction, respectively. The PO-TMB reaction product was stabilized effectively by the addition of sodium tungstate to the reaction mixture, thus making the PO-TMB reaction now generally applicable to in situ hybridization (ISH). To avoid mixing of the precipitates of the two PO reactions used in the triple-color ISH method, the first detected PO activity was always completely inactivated by a mild acid treatment before the second one was applied. Finally, the cell preparations were embedded in a thin protein layer cross-linked by formaldehyde to ensure permanent stabilization of the enzyme reaction products and optimal visualization of color contrast. The triple-color ISH detection procedure could be combined with beta-galactosidase-5-bromo-4-chloro-3-indolyl-beta- D-galactoside (beta-Gal-BCIG) immunocytochemistry (ICC), leading to the simultaneous localization of multiple DNA targets and a protein target in the same cell. The described procedure may therefore be a valuable tool in the areas of cytogenetics, cell biology, and molecular pathology.
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PMID:A novel triple-color detection procedure for brightfield microscopy, combining in situ hybridization with immunocytochemistry. 793 May 13

The lungs of rats exposed to formaldehyde vapor, for 6 hr/day over 4 consecutive days, were examined for signs of injury and for changes in the level, or activity, of cytochrome P450. The animals were supplied with 10 ppm formaldehyde vapor generated, in two separate experiments, either from an aqueous solution of formaldehyde or from heated paraformaldehyde. All rats were exposed for 6 hr, on each of 4 consecutive days, and killed 1 day after the onset of the fourth period of exposure. The lung weights and gains in body weight of exposed animals were indistinguishable from those of their controls. Lungs from the formaldehyde-exposed animals did not show any signs of injury, even at the ultrastructural level. Bronchoalveolar lavage samples from exposed animals showed no increase in alkaline phosphatase or gamma-glutamyl transpeptidase activity. The total concentration of cytochrome P450 in the lungs of exposed animals was similar to that found in their controls. The P450 activity of pulmonary microsomes from exposed animals was not significantly different from that obtained with samples from the control animals. These results indicate that repeated exposure to 10 ppm formaldehyde vapor does not injure the deep lung of rats and has no effect on the level of lung P450 or on its activity against substrates for the most common pulmonary forms of this enzyme.
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PMID:Pulmonary cytochrome P450 in rats exposed to formaldehyde vapor. 810 Jul 69


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