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)

A method using p-benzoquinone for coupling antigens and antibodies to enzymes and erythrocytes is described. The method involves the treatment of proteins (or polysaccharides) at pH 6 or 7 with an excess of p-benzoquinone. After removal of the unreacted reagent by gel filtration, the "activated" proteins were coupled at pH 8-9 with enzymes or erythrocytes. Biological activities of the proteins were not substantially modified by this treatment since 80-100% of the antigen binding capacity was found to be preserved in p-benzoquinone treated antibodies or Fab fragments. Anti-Ig antibodies (or Fab) were coupled by this procedure to peroxidase, alkaline phosphatase, lactoperoxidase, glucose oxidase and beta-galactosidase, and the conjugates obtained were found to be highly effective in detecting intracellular Ig by immunohistochemical techniques. Erythrocytes coated with sheep anti-mouse Ig antibody or Fab were used to titrate by passive hemagglutination serum Ig. The same erythrocytes were employed to detect by plaque assay mouse Ig secreting cells. Erythrocytes coated with peroxidase, alkaline phosphatase, bovine serum albumin, ribonuclease, Salmonella polysaccharide (B 27 +) and pneumoccocal polysaccharide SIII were employed to titrate serum antibody by passive hemagglutination and hemolysis and to detect mouse antibody secreting cells by plaque assay. All the antigens and antibodies coated erythrocytes prepared gave highly satisfactory and reproducible results.
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PMID:A new method using p-benzoquinone for coupling antigens and antibodies to marker substances. 0 79

The aim of the present investigation was to test a procedure useful for the electron-microscopic in situ identification of the presumptive cariogenic microorganism Streptococcus mutans (serotype d) growing in the human dental plaque. For this purpose, different parameters of an indirect immunohistochemical method were tested in three sets of experiments. In experiment set I, all serological and histochemical procedures were performed en bloc on specimens fixed only with glutaraldehyde before embedding in Vestopal W. Different marking substances, such as ferritin, alkaline phosphatase and horseradish peroxidase (HRP) were tested. The en bloc method using HRP-labelled antibodies was found useful for staining of in vitro-grown bacteria but failed when applied to dental plaque. In experiment set II, ultrathin sections of glutaraldehyde-fixed and glycol methacrylate-embedded in vitro-grown bacteria were section-stained. The bacteria became outlined with a highly electron-dense HRP reaction product which accumulated on top of the cell envelope. The same type of HRP reaction product was found on some bacteria in the 2-day-old dental plaque after section-staining. This system was further tested in a series of controls also using consecutive serial sections. In exp. set III, a number of different stationary and transient oral bacteria were immunohistochemically section-stained. A cross-reaction with bacteria belonging to S.salivarius was discovered and removed by absorbing the anti-S.mutans serotype d serum with S.salivarius (NCTC 8618).
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PMID:Some immunohistochemical experiments aiming at the electron-microscopic in situ identification of a dental plaque microorganism--Streptococcus mutans. 8 15

In the course of serial passages for several years a line of uncloned HeLa cells (A line) showed a gradual decrease in plaquing efficiency by coxsackievirus A9 (CA9 virus), while subcultures prepared from the same line kept frozen at an early passage level (A original line) did not show any change. However, it was observed later that the plaque-forming ability of the A original line (A orig. line) also decreased after serial passages as was observed with the A line. Comparing the characteristics of the same cell line at two different passage levels, it was found that the efficiency of adsorption of virus to cells was nearly the same, while virus yield at 8 hours after infection was different. The activity of alkaline phosphatase of cells was also different between these two passage levels, suggesting that a high enzymatic activity is associated with the susceptibility of cell cultures to CA9 virus. Magnesium chloride at 25 mM enhanced plaque formation by CA9 virus in highly passaged less susceptible cell cultures, and a possible role of the chemical as a stabilizer of alkaline phosphatase was discussed.
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PMID:Variation in susceptibility of HeLa cell lines to coxsackievirus A9. 22 35

The carotid artery of the rabbit is a suitable blood vessel to study radiation induced atheromatosis in hypercholesteremic animals, because no plaque formation occurs within two months after the start of a 0.5% cholesterol diet. Cholesterol contents as high as 2% however, do give atheromatous plaques in the carotid artery without prior irradiation. As early as five hours after local irradiation of the carotid artery activation of the plasma membrane-bound enzyme alkaline phosphatase could be observed in some intimal cells. Two to three days after irradiation the activity disappeared. This phenomenon was observed in normo-and hypercholesteremic irradiated arteries. Depending on the lipid content of the blood, infiltration of lipids was observed at one day after the irradiation or later, accompanied by activation of beta-glucuronidase in the innermost layer of medial cells. Hereafter plaque formation started and cell proliferation could be found in the subendothelial space. It is assumed that because of the irradiation, the endothelial cells of the carotid artery are damaged in such a way that they do not function properly as a barrier against lipoprotein entrance from the blood into the arterial wall. The lipid infiltration caused lysosomal activation and probably cellular proliferation.
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PMID:Initial events in radiation-induced atheromatosis. II. Damage to intimal cells. 71 13

Peroxidase (PO), alkaline phosphatase, and glucose oxidase, as well as Fab anti-PO, were coupled with varying molar ratios of trinitrophenyl (TNP) hapten. These reagents were evaluated for their ability to detect anti-TNP antibodies in the lymph node cells of Balb/c mice immunized with heavily-substituted TNP45 alkaline phosphate, which gave rise only to anti-hapten antibody-forming cells (AFC). The best results were obtained with lightly-substituted TNP-Fab anti-PO plus PO, TNP-alkaline phosphatase, and TNP-glucose oxidase. These reagents gave strong, specific staining of AFC, and negative background staining. Anti-hapten and anti-carrier AFC could be stained in contrasting colors on the same slide, when immunization was performed with lightly-substituted TNP5.9 alkaline phosphatase. Anti-hapten AFC were detected with TNP-Fab anti-PO or TNP-glucose oxidase, and unsubstituted alkaline phosphatase was used to reveal anti-carrier AFC. The number of AFC detected with these reagents was compared with the number of direct and indirect anti-TNP plaque-forming cells (PFC). At three and five weeks after primary immunization, 40 and 70% more AFC than PFC were detected. These methods can be employed alone, to enumerate anti-TNP AFC and, if desired, anti-carrier AFC; they can also be used in parallel with anti-TNP PFC assays, to determine the fractions of AFC that are not actively involved in antibody secretion.
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PMID:Detection of anti-TNP antibody-forming cells (AFC) with TNP-enzyme and TNP-Fab anti-enzyme conjugates. 77 69

Thirty five patients with psoriasis (plaque type 26, guttate 3, pustular 4, and erythrodermic 2) were treated with oral mycophenolic acid for a period ranging from 52 to 104 weeks. The average follow-up was 89 weeks, and the dose schedule ranged from 2,400 to 7,200 mg daily. Excellent response was noted in 20 patients, good in 13 patients, and poor in 2. The most common clinical side effects were in the gastrointestinal tract, namely, diarrhea, nausea, abdominal cramps, and soft stools. A high incidence of herpes simplex, herpes zoster, and a flu-like syndrome was noted. Laboratory abnormalities consisted of mild blood hemoglobin reduction, one case of leukopenia (3,9000 WBCs per cubic millimeter), two cases with thrombocytopenia and mild elevation of alkaline phosphatase. Mycophenolic acid appears as a promising drug for the treatment of severe psoriasis.
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PMID:Mycophenolic acid in the treatment of psoriasis: long-term administration. 87 14

Helicobacter pylori (H.p.) is a microorganism involved in peptic ulcer disease. To clarify the role of human dental plaque as a reservoir of H.p. and to compare different methods of investigation the authors studied 20 patients, 17 males main age 56 +/- 12 and 3 females 52 +/- 7, gastro-duodenal H.p. positive. The trial was carried out by cultural, biochemical and microscopical plaque analysis. Cultural and microscopical method were H.p. positive in 80% patients, urease in 100%, alkaline phosphatase in 80%, gamma glutamyltransferase in 70%, nitrate in 70%. To minimize the possibility of false results in H.p. plaque analysis it is necessary to use the three methods simultaneously. Further trials both on human plaque and on food and beverages will be useful to clarify the role of H.p. in human pathology.
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PMID:Comparison of three different methods for evaluation of Helicobacter pylori (H.P.) in human dental plaque. 130 23

The relationship between the level of urokinase-type plasminogen activator (uPA) and pelvic lymph node metastasis was investigated in 20 patients with invasive cervical cancer of the uterus. Frozen sections from all surgical specimens were stained immunohistochemically by alkaline phosphatase anti-alkaline phosphatase method to detect uPA in cancer tissue. The concentration of uPA, determined immunologically, and the fibrinolytic activity were also examined in supernatants of homogenates of some cancer tissues. uPA was detected immunohistochemically in cancer cells of all specimens. A significant correlation was found between the extent of immunohistochemical staining for uPA and the concentration of uPA determined immunologically in cancer tissues (P less than 0.05). A positive correlation was also found between semiquantitative values determined by immunohistochemical staining for uPA and lymph node metastasis (P less than 0.05). Fibrinolytic activity in cancer tissues was confirmed by casein-plaque assay. These findings indicate that the pro-uPA/uPA contents in cervical cancer tissues are clinically useful for predicting the metastatic potential of these cancers to the pelvic lymph nodes.
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PMID:Clinical significance of urokinase-type plasminogen activator (uPA) in invasive cervical cancer of the uterus. 152 11

This study evaluated the feasibility of using a colony lift method and DNA probes to enumerate bacterial species cultured on primary isolation plates. Fourteen digoxigenin-labeled whole chromosomal DNA probes representing 12 subgingival species were validated by hybridization with colony lifts prepared from 249 reference strains of 51 species grown on Trypticase soy agar plates supplemented with 5% sheep blood. Colonies of reference strains were lifted onto Nytran filters from plates and treated to lyse cells, remove cellular proteins, denature and fix microbial DNA to the filters. Positive reactions were detected with an anti-digoxigenin antibody conjugated to alkaline phosphatase and revealed by bromo-chloro-indolyl phosphate and nitroblue tetrazolium. Cross-reactions were not observed for 13/14 probes, but 2 strains of Streptococcus mitis reacted with the probe to Streptococcus sanguis II. Subgingival plaque samples were taken by means of a sterile curette from mesiobuccal surfaces of teeth present in each of 26 subjects with differing periodontal disease states. Samples were dispersed, diluted, plated and incubated anaerobically for 7 d at 35 degrees C. Colonies were lifted as described above. Filters were cut into sections and hybridized with the 14 digoxigenin-labeled DNA probes. The probes were used to enumerate the test species and the total number of isolates was determined in 711 plaque samples. The colony lift method and DNA probes provided a sensitive, economical and quantitative method for enumerating cultivable microbial species in subgingival plaque samples. In addition, the amplification provided by growing the organisms on agar plates facilitated determination of numbers of organisms in small plaque samples, such as those from healthy sites.
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PMID:Enumeration of subgingival species on primary isolation plates using colony lifts. 152 19

Nonisotopic, whole-genomic DNA probes, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), biochemical tests in microtiter trays and cellular fatty acid (CFA) analysis were compared for the identification of 5 oral Selenomonas species. DNA probes were prepared by biotin-labeling DNA extracted from the type strains of Selenomonas noxia, Selenomonas flueggei, Selenomonas artemidis, Selenomonas infelix and Selenomonas sputigena. The probes were hybridized with DNA from 21 reference strains, 18 fresh isolates of Selenomonas species, and 21 strains of other oral gram-negative species. Target DNAs were obtained by in situ extraction of colonies blotted onto filter paper. Streptavidin-linked alkaline phosphatase was used to detect homologous reactions of probe and target DNA. Each Selenomonas species DNA probe reacted with reference strains of only that species. All Selenomonas strains that reacted with the DNA probe for a particular species gave similar biochemical test results, SDS-PAGE protein profiles, and CFA profiles to those of the type strain of the corresponding species. All the methods tested were useful for identifying the species, and all yielded similar identifications of the fresh isolates. The DNA probes, however, had the potential for identifying Selenomonas species directly from primary isolation plates or plaque samples.
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PMID:Identification of Selenomonas species by whole-genomic DNA probes, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, biochemical tests and cellular fatty acid analysis. 152 28


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