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)

The 10 S DNA polymerase alpha from calf thymus (Masaki, S., and Yoshida, S. (1978) Biochim. Biophys. Acta 521, 74-88) has been purified to near homogeneity. The most purified fraction obtained by repeated sucrose rate-zonal centrifugation contained three large polypeptides of 150,000, 145,000, and 140,000 daltons and three to four smaller polypeptides ranging from 43,000 to 50,000 daltons. A good resolution of these polypeptides was achieved on a sodium dodecyl sulfate-polyacrylamide linear gradient gel (5-20%) which was stained by the silver stain method. The three large polypeptides were also observed in the more crude fractions prepared in the presence of three kinds of protease inhibitors. By a peptide mapping analysis, it was revealed that these three polypeptides have a similar primary structure. Treatments of the enzyme with alkaline phosphatase, phosphodiesterase, and neuraminidase did not affect the gel pattern. These results indicate that the 10 S DNA polymerase alpha of calf thymus has a microheterogeneity in terms of the large polypeptide component. Among these three large polypeptides, the two polypeptides of 150,000 and 145,000 daltons disappeared by keeping the sucrose gradient fraction at 4 degrees C in the absence of glycerol, while the 140,000-dalton polypeptide was well preserved. The poly(rA)oligo(dT)-dependent activity of 10 S DNA polymerase alpha was selectively lost under this condition.
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PMID:10 S DNA polymerase alpha of calf thymus shows a microheterogeneity in its large polypeptide component. 708 21

This study was designed to determine the potential benefit or toxicity of an immunomodulator, Corynebacterium parvum vaccine, when it is given after severe burn injury. Forty conditioned beagles received a 33% total body surface 3-degree flame burn and were resuscitated with Ringer's lactate solution (3 ml/kg/% burn). Wounds were treated daily for 10 days with silver sulfadiazine cream. Two days and nine days after burn, 21 of the animals received C. parvum vaccine (10 mg/kg IV) in a saline infusion, while 19 control animals were given only saline infusion according to a double-blind protocol. Serial measurements were made of temperature, weight, food intake, hematocrit, hemoglobin, red blood count, white blood count, differential, platelet count, fibrin degradation products, activated partial thromboplastin time, clot retraction, C3, blood cultures, neutrophil function, monocyte function, opsonic index, Na, K, Cl, BUN, glucose creatinine, total protein albumin, albumin/globulin ratio, alkaline phosphatase, SGPT, and SGOT. During 45 days of observation, only 16% of the saline control dogs survived compared to 47% of the treated animals. Total white counts and neutrophil function were the only values which were significantly better in animals receiving C. parvum. However, their correlation with increased survival was marginal This preclinical trial suggests that C. parvum is an effective immunodulator for prevention of fatal infection following burn injury. There were no demonstrable toxic effects of the material in this study.
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PMID:Improved survival in severely burned animals using intravenous Corynebacterium parvum vaccine post injury. 745 9

The expression of transforming growth factor beta 1 (TGF-beta 1) mRNA has been determined in 16 breast carcinomas using in situ hybridization and compared with TGF-beta protein as detected by antibodies against TGF-beta 1 and TGF-beta 1 plus TGF-beta 2. Digoxigenin-labelled riboprobes were used, with alkaline phosphatase and immunogold silver detection systems. TGF-beta 1 mRNA was only detected in carcinomas in which TGF-beta 1 protein was found (9 of 16 cases) and not in those with prominent reactivity for TGF-beta 2. RNA preservation was poor in two other cases in which TGF-beta 1 protein had been detected. In general, those tumours with greater numbers of cells labelled for TGF-beta 1 mRNA had prominent reactivity for TGF-beta 1 protein. The mRNA was localized to cancer cells with no labelling of stromal cells, although in a small number of cases scanty staining for TGF-beta 1 protein had been observed in stromal cells. The incidence of detection of TGF-beta 1 mRNA is lower than the published data from Northern analysis studies of breast carcinomas, suggesting that only higher levels of TGF-beta 1 mRNA expression are being detected by in situ hybridization. However, this approach has provided useful information about the cellular sites of expression of TGF-beta 1 in breast carcinomas.
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PMID:Determination of transforming growth factor beta 1 mRNA expression in breast carcinomas by in situ hybridization. 749 Jun 77

We present a method for an epi-illumination immunohistochemical double staining approach. The method combines the use of an immuno-alkaline phosphatase technique and the immunogold-silver technique, visualized with epifluorescence and epi-polarization illumination, respectively. Out of six tested alkaline phosphatase activity-revealing methods, only the reaction product obtained with the Becton Dickinson CAS Red kit showed an intense red fluorescence with a rhodamine filter set and no signal with epi-polarization illumination. The silver precipitate did not exhibit any signal with the rhodamine filter set. This allows separate observation and photographic recording of two antigens in one tissue section, an objective that cannot be achieved with conventional immunoenzyme double staining methods. The double epi-illumination approach presented is compatible with different immunoenzyme double staining protocols.
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PMID:Double epi-illumination microscopy with separate visualization of two antigens: a combination of epi-polarization for immunogold-silver staining and epi-fluorescence for alkaline phosphatase staining. 750 69

Double immunoenzymatic labelling procedures for the localization of antigens on cells in tissue sections using horseradish peroxidase (HRP) and alkaline phosphatase have been described previously, but mainly for detecting antigens on different cells. With this type of staining when two antigens are present on the same cell, an optimal colour combination that shows a high contrast between the basic colour of each enzyme substrate product is difficult to achieve and the interpretation of their mixed colour intermediate is subjective. We present a method for the simultaneous demonstration of two antigens on the same cell. The method can be used to label either single cells in suspension, or cells in paraffin fixed tissue, using a combination of a particulate label, colloidal immunogold-silver, and an enzymatic label HRP-DAB. The method is easy to perform and utilises commercially available staining kits.
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PMID:Dual staining of lymphocyte membrane antigens with colloidal gold and biotinylated horseradish peroxidase. 750 81

Monoclonal antibodies (mAbs) specific for cytokeratins are potent probes for the identification of disseminated individual epithelial tumour cells in mesenchymal organs such as bone marrow. We have used a monoclonal antibody (mAB) against cytokeratin 18 (CK18) for the detection of individual metastatic tumour cells in bone marrow aspirates from 84 patients with carcinoma of the prostate. CK18+ cells were detected in a sensitivity of 1 per 8 x 10(5) marrow cells using the alkaline phosphatase anti-alkaline phosphatase (APAAP) system for staining. We were able to detect CK18+ tumour cells in the marrow of 33% of patients with stage N0M0 prostate cancers. The incidence of CK18+ cells showed a significant correlation with established risk factors, such as local tumour extent, distant metastases and tumour differentiation. For further characterization of such cells in patients with prostate cancer, we developed an immunocytochemical procedure for simultaneous labelling of cytokeratin component no. 18 (CK18) and prostate-specific antigen (PSA). In a first step, cells were incubated with a murine mAb against PSA, followed by gold-conjugated goat anti-mouse antibodies. In a second step, a biotinylated mAb to CK18 was applied as primary antibody and subsequently incubated with complexes of streptavidin-conjugated alkaline phosphatase, which were developed with Newfuchsin substrate. The binding of gold-labelled antibodies was visualized by silver enhancement. CK18+ cells co-expressing PSA were found in bone marrow aspirates from 5 out of 14 patients with carcinomas of the prostate. The specificity of CK18 for epithelial tumour cells in bone marrow was supported by negative staining of 12 control aspirates from patients with benign prostatic hyperplasia (BPH).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Immunocytochemical detection and phenotypic characterization of micrometastatic tumour cells in bone marrow of patients with prostate cancer. 752 Oct 88

The feasibility of various non-isotopic enzymatic detection systems was tested for in situ hybridization using biotin-labelled, nick-translated cDNA probes. For this purpose, we isolated and prepared cDNA restriction fragments encoding the proteolytic cysteine proteinase cathepsin L and analysed Kirsten murine sarcoma virus-transformed BALB/3T3 cells, which have been shown to express high amounts of cytoplasmic RNA of this ras oncogene-induced proteinase. When compared on a semiquantitative basis, colorimetric non-isotopic detection of cDNA hybrids with avidin-biotin-peroxidase conjugates visualized by silver intensification of the nickel-diaminobenzidine end-product was superior to that obtained with avidin-biotin-alkaline phosphatase using different substrates for development. When the peroxidase staining technique was applied for RNA detection, it was found that overnight incubation in methanol containing hydrogen peroxide followed by deproteination with HCl was the most effective method for inhibition of endogenous peroxidase activity. For DNA detection, non-specific nucleic staining was completely abolished when heat treatment (100 degrees C) of the cell specimens was performed prior to hybridization.
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PMID:Enzymatic detection systems for non-isotopic in situ hybridization using biotinylated cDNA probes. 763 60

Early dissemination of malignant cells is the main cause for metastatic relapse in patients with solid tumours. By use of monoclonal antibodies (mAbs) specific for cytokeratins, disseminated individual epithelial tumour cells can now be identified in mesenchymal organs such as bone marrow. Further to characterize such cells in patients with prostate cancer, an immunocytochemical procedure was developed for simultaneous labelling of cytokeratin component no. 18 (CK18) and prostate specific antigen (PSA). In a first step, cells were incubated with mAb ER-PR8 against PSA and secondary gold-conjugated goat anti-mouse antibodies. In a second step, biotinylated mAb CK2 to CK18 was applied as primary antibody and subsequently incubated with complexes of streptavidin-conjugated alkaline phosphatase, which were developed with the Newfuchsin substrate. The binding of gold-labelled antibodies was visualized by silver enhancement. The sensitivity and specificity of the technique was demonstrated on cryostat sections of hyperplastic prostatic tissue, and cytological preparations of LNCaP prostatic tumour cells. Double staining was restricted to cells derived from the secretory epithelium of the prostate. Cross-reactivity between both detection systems was excluded by several controls, including the use of unrelated antibodies of the same isotype and the staining of CK18+/PSA- HT29 colon carcinoma cells. CK18+ cells co-expressing PSA were found in bone marrow aspirates from 5 out of 13 patients with carcinomas of the prostate, a finding that is consistent with the relative fraction of double-positive LNCaP cells. The specificity of CK18 for epithelial tumour cells in bone marrow was supported by negative staining of 12 control aspirates from patients with benign prostatic hypertrophy.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Immunocytochemical double staining of cytokeratin and prostate specific antigen in individual prostatic tumour cells. 768 10

The breakthrough of chemiluminescence in the field of solution immunoassays and transfer membranes prompted us to explore whether a light-based detection system could provide a gain in sensitivity over chromogenic and FITC markers for nucleic acid and protein detection on histological preparations. A Hamamatsu device and an enhanced chemiluminescence (ECL) luminol substrate of the peroxidase were used to detect epithelial and endothelial components by immunohistochemistry (IHC) and for in situ hybridization (ISH) of papilloma virus DNA. The accuracy of the signal was compared to that obtained with DAB-peroxidase, silver-enhanced DAB-peroxidase, NBT-BCIP-alkaline phosphatase, and FITC. Our results demonstrated the feasibility and high sensitivity of luminescence detection for histological preparations. In part due to the ultrasensitive videocamera and photon-counting imaging, interpretable and reproducible results were obtained within counting times shorter than 5 min, and with dilutions of the primary antibodies 100- to 10,000-fold greater than those used for chromogenic and FITC reactions. As for ISH, with identical concentrations of the HPV 18 DNA probe on HeLa cells, labeling was apparent by luminescence but undetectable with the chromogen. The morphological resolution allowed a discriminatory analysis of the signal. Therefore, at the light microscopic level, enhanced chemiluminescence offers an appealing alternative to FITC and chromogenic markers for detection and quantification of low-concentration molecules.
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PMID:Enhanced chemiluminescence: a high-sensitivity detection system for in situ hybridization and immunohistochemistry. 769 29

We report on four patients of our own and another thirty-six from the literature, who developed almost identical and unusual clinical syndromes after surgical treatment of hydatid disease of the liver, with the aim of showing the extremely serious nature of the problem that can ensue. An association of four factors seems to be necessary to promote caustic sclerosing cholangitis: a) injection of a scolicidal agent (formalin, hypertonic saline, ethanol, silver nitrate or iodine solution) into the cyst cavity; b) a communication between the cyst and the biliary tree; c) a condition that prolongs the exposure of the biliary tree to the scolicidal; and d) a particular sensitivity to the scolicidal agent. While this last condition cannot be anticipated, we may justifiably conclude that surgeons should not inject a scolicidal solution into the hydatid cyst, but prevent intra-abdominal diffusion of the parasite by using hydrogen peroxide, gauze pads moistened by a scolicidal solution or by preoperative chemotherapy with albendazole. Caustic sclerosing cholangitis has an earlier onset of symptoms and a more rapidly progressive nature than primary sclerosing cholangitis. In foresight, serum alkaline phosphatase should be monitored and, when raised, a retrograde endoscopic cholangiogram and/or a liver biopsy should be performed. Digestive shunt surgery should be avoided and the possibility of liver transplantation has to be periodically evaluated.
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PMID:Caustic sclerosing cholangitis. Report of four cases and a cumulative review of the literature. 785 56


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