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 relatively simple, very sensitive bioluminescence-enhanced detection system for protein blots is described. The method utilizes antibodies conjugated with alkaline phosphatase. The alkaline phosphatase then takes part in a reaction by releasing D-luciferin (Photinus pyralis) from D-luciferin-O-phosphate. Liberated D-luciferin reacts with luciferase, ATP and oxygen with light emission. The light is detected by a sensitive photographic film, thereby permitting the visualization of the alkaline phosphatase-conjugated antibodies. Under non-optimized conditions the limit of detection is at present 5 to 50 pg of protein, corresponding e.g. to 30 to 300 x 10(-18) mol of rabbit immunoglobulin G. The detection system is therefore 100 times more sensitive than other systems used at present.
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PMID:A new, very sensitive, bioluminescence-enhanced detection system for protein blotting. Ultrasensitive detection systems for protein blotting and DNA hybridization, I. 369 39

The use of gas chromatography-mass spectrometry (GC-MS) for characterization of free radical-induced base damage to DNA is presented. Damage introduced to DNA by reactive oxygen species such as hydroxyl radicals appears to play an important role in mutagenesis, carcinogenesis and aging. Elucidation of the chemical nature of such DNA lesions is necessary for the assessment of their biological consequences and enzymatic repair. DNA exposed to radiation-generated hydroxyl radicals in aqueous solution was hydrolyzed to 2'-deoxyribonucleosides with a mixture of DNase I, venom and spleen exonucleases and alkaline phosphatase. The hydrolysate was subsequently trimethylsilylated and analyzed by GC-MS. A large number of DNA lesions were separated and identified. Mass spectra obtained were interpreted on the basis of the typical fragmentation pathways of trimethylsilylated nucleosides. The use of GC-MS with selected-ion monitoring facilitated the detection of these lesions at the very low quantities and radiation doses (below 10 Gray) that might be relevant to those in biological systems.
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PMID:Characterization of free radical-induced damage to DNA by the combined use of enzymatic hydrolysis and gas chromatography-mass spectrometry. 378 50

Changes in three recognized liver function tests are reported following the use of propofol in 30 fit, unpremedicated women in whom propofol was used as the main anaesthetic agent. Doses of 140 to 330 mg were given, together with nitrous oxide and oxygen. All patients were undergoing minor gynaecological operations and all conformed to Grade 1 physical status of American Society of Anesthesiologists Classification. In none of these patients was there hypoxia or hypercarbia at any time during or following anaesthesia and none of the patients received any other drugs until completion of the study. No significant changes in liver enzymes (aspartate transaminase and alanine transaminase) or in serum alkaline phosphatase were detected.
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PMID:Changes in liver function tests after propofol ('Diprivan'). 387 86

31P NMR signals from substrates and products of alkaline phosphatase have been adapted to measure the rates and product ratios for the hydrolysis and phosphotransferase reactions from pH 6 to 10. Below pH 8, glycerol is a poorer acceptor than H2O (glycerol phosphates:Pi = 0.5). Tris is a more effective acceptor below pH 8, showing a maximum acceptor efficiency at pH 8 (Tris phosphate:Pi = 2). Phosphotransferase efficiencies are in the order expected for the pKaS of the alcohol groups, Tris less than glycerol Cl, C3 less than glycerol C2. Tris and glycerol induce chemical shifts in 113Cd(II) present at the A site but not the B or C sites of the metal triad present at each active center of Cd(II)6 alkaline phosphatase, suggesting that the alcoxides of the acceptors coordinate the A site metal and become the nucleophiles attacking the phosphoseryl residue (E-P) in the second step of the mechanism. The interaction is through the oxygen of Tris. The transferase activity of the amino alcohol shows a bell-shaped pH dependency. Aliphatic alcohol acceptors show small increases in acceptor activity between pH 6 and 8, with 5-fold increases from pH 8 to 10 (at pH 10, glycerol phosphates:Pi = 2.5). 31P NMR inversion transfer has been used to measure the koff for Pi dissociation from the noncovalent enzyme complex (E . P). For the Zn(II)4 alkaline phosphatase koff is essentially pH independent at approximately 35 s-1. For Cd(II) or Mg(II) at the B site in place of Zn(II), koff less than or equal to 1 s-1 X Cl-ion, which appears to coordinate the A site metal ion, enhances koff, suggesting that both Cl- and HPO2-4 can coordinate the A site metal ion in a 5-coordinate intermediate. pH control of the alkaline phosphatase mechanism appears to reside in the stability of E-P and not the dissociation of E . P, compatible with the hypothesis that the activity-linked pKa is that of a H2O molecule coordinated to the A site metal, which in the hydroxide form becomes the nucleophile attacking the phosphoseryl group (E-P).
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PMID:Alkaline phosphatase. 31P NMR probes of the mechanism. 388 2

Three groups of patients received Althesin, minaxolone or di-isopropyl phenol to supplement 67% nitrous oxide in oxygen. A fourth group receiving halothane to supplement nitrous oxide in oxygen acted as a control. Hepatic function tests were measured before operation and on days 1, 3, 5 and 7 after major vascular reconstructive surgery. There were significant increases to a mean value above the upper limit of normal in aspartate amino-transferase activity by day 3 in all groups. Total lactic dehydrogenase activity increased in the patients receiving Althesin, minaxolone and halothane. No change was seen in the alkaline phosphatase in any of the study groups. Gamma glutamyl transpeptidase increased in all groups, but the mean value at day 7 was not greater than the upper limit of normal. The mean activity of ornithine carbamoyl transpeptidase showed no change in any group throughout the study period. Two of the patients receiving minaxolone suffered cholestatic jaundice during the first month. These results suggest that anaesthesia with Althesin or di-isopropyl phenol results in enzyme changes similar to those seen in a comparable group of patients receiving halothane to supplement nitrous oxide in oxygen anaesthesia.
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PMID:Hepatic function after anaesthesia for major vascular reconstructive surgery. 613 33

The sites of cleavage of DNA by bleomycin A2, bleomycin B2, phleomycin, tallysomycin A, and Blenoxane (Bristol-Meyers) in reactions containing equimolar Fe2+ and atmospheric oxygen were analyzed by gel electrophoresis of 32P end labeled DNA fragments. Bleomycin A2 and bleomycin B2 reactions cleaved DNA at all sites with a frequency equal to that of Blenoxane. At high concentrations of bleomycin the site specificity of cleavage was unchanged. Bleomycin cleavage sites and phleomycin cleavage sites are a subset of sites cleaved in reactions containing tallysomycin A. The nature of 5' and 3' termini induced by bleomycin cleavage was investigated. Electrophoresis of bleomycin-induced fragments after alkaline phosphatase or polynucleotide kinase treatment indicated that 5' termini are phosphoryl groups but 3' termini are not simple phosphoryl groups. Analysis of bleomycin cleavage of single-stranded DNA substrate showed that cleavage occurs only in regions of potentially double-stranded looped-back sequences. Possible mechanisms for determination of bleomycin cleavage sequence specificity are discussed.
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PMID:Specificity of deoxyribonucleic acid cleavage by bleomycin, phleomycin, and tallysomycin. 618 7

In 38 patients subjected to retropubic prostatectomy the effects of continuous lumbar epidural analgesia for 24 hours and the thiopentone- oxygen-nitrous oxide- alcuronium-pethidine sequence with artificial ventilation on the serum activities of aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), alpha-hydroxybutyrate dehydrogenase (HBD), and alkaline phosphatase (AP) have been studied. Per- and postoperative complications were recorded according to a prearranged plan designed to quantify the peroperative haemorrhage, postoperative deep vein thrombosis, pulmonary, circulatory and infectious complications. ASAT, ALAT and AP in the general group and ALAT in the epidural group showed significant increases on the 5th and 7th postoperative days. There existed no statistically significant difference between the groups. 82% of the patients with documented postoperative complications combined with hypoxaemia showed a pathologic liver enzyme pattern in contrast to 9% of the patients with uneventful postoperative course. It is concluded that the method of anaesthesia did not have an effect on the liver enzymes. Complications combined with postoperative hypoxaemia seemed to be the factors responsible for the increases of liver enzymes.
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PMID:Liver enzymes after retropubic prostatectomy in patients receiving continuous lumbar epidural analgesia or general anaesthesia. 618 33

Oxygen and glucose consumption and lactate production of the peritoneal membrane and intra-abdominal adhesions were measured in rats after a single intra-peritoneal colloidal silica injection. Enzyme histochemical studies were made of lactate dehydrogenase, succinate dehydrogenase, NADH2-diaphorase, NADPH2-diaphorase, glucose-6-phosphate dehydrogenase, glutamate dehydrogenase, acid phosphatase, leucylaminopeptidase and alkaline phosphatase in the peritoneal membrane. Anaerobic glycolysis comprises 47% of the total glucose consumption in the the normal peritoneum. Glucose consumption and lactate production of the peritoneal membrane increased sharply in the early phase of silica-induced peritonitis and stayed at a high level for a week indicating an enhanced anerobic metabolism. Oxygen and aerobic glucose consumption increased more slowly than anaerobic glucose consumption and reached their maxima 1 week after silica injection, indicating that the rate of aerobic metabolism is also higher in chemical peritonitis than in the controls. On the other hand, glucose consumption and lactate production increased in a parallel fashion in adhesions and in the peritoneum in the early phase of peritonitis. However, the maximum and later levels were less in adhesions than in the peritoneum. In the enzyme histochemical study high activities of enzymes indicating anaerobic energy metabolism and metabolism via the pentose phosphate shunt were seen in cells of the peritoneal membrane during the early phase of peritonitis. No activity was identified in enzymes indicating aerobic energy metabolism and increased catabolism before the end of the first week.
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PMID:Energy metabolism of the peritoneal membrane in silica-induced peritonitis. A biochemical and enzyme histochemical study. 625 64

Alteration of the surface of human neutrophils with the nonpenetrating, protein-inactivating agent p-diazobenzenesulfonic acid (DASA) was found to prevent activation of the respiratory burst by some stimuli, but not others. Production of superoxide anion (O2-) stimulated by concanavalin A or the chemotactic peptide formyl-methionyl-leucyl-phenylalanine FMLP was inhibited by DASA pretreatment, whereas O2- production stimulated by phorbol myristate acetate (PMA), sodium fluoride. or the ionophore A23187 was not inhibited by DASA. Pretreatment with DASA inhibited oxygen uptake stimulated by FMLP, but not oxygen uptake stimulated by PMA. DASA reproducibly inhibited activities of two known surface enzymes Mg++-ATPase and alkaline phosphatase, by 45-55% and 60-70%, respectively. The inhibition by DASA of O2- production did not appear to be caused by interference with binding of the affected stimuli, since pretreatment with DASA did not inhibit release of the lysosomal enzymes lysozyme and myeloperoxidase induced by concanavalin A or FMLP. Membrane-rich particulate fractions from neutrophils have been shown to contain NADPH-dependent oxidative activity that is presumably responsible for the phagocytosis-associated respiratory burst of intact cells. The PMA-activated enzyme was susceptible to inhibition of directly exposed to DASA in this particulate fraction. These findings suggest that more than one mechanism exists for activation of the respiratory burst oxidase in human neutrophils, and that the neutrophil possesses at least one oxidase that is not an ectoenzyme.
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PMID:Respiratory burst enzyme in human neutrophils. Evidence for multiple mechanisms of activation. 625 8

In order to determine the substrate specificity of the alkaline phosphatase (APase) which appears in murine lymphomas, two substrates were chosen. p-Nitrophenyl phosphatase (pNPP), the standard substrate, has an oxygen-phosphorus (O-P) bond. Cysteamine-S-phosphate (CASP) has a sulfur-phosphorus (S-P) bond. It has been reported that murine lymphoma APase does not cleave the S-P bond of CASP. These results were not confirmed. Under all conditions tested, the murine lymphoma APase showed consistent hydrolysis of CASP at approximately one-third the rate of hydrolysis of pNPP. Biochemical characteristics used to confirm this include pH optimum, heat inactivation, kinetics, magnesium activation, L-homoarginine inhibition, EDTA inhibition, and activity remaining during stages of partial purification. It is concluded that the murine lymphoma APase shows a preference of pNPP as a substrate but does hydrolyse CASP at a lower rate. The conflict between our laboratory and that of Neumann's may be one of interpretation rather than data. Neumann shows similar data to ours in terms of the 3-fold ratio, but considers all hydrolysis of CASP as representative of 'normal' APase. She uses a formula to calculate the amount of lymphoma APase based on the ratio 1.9 which is the ratio of hydrolysis of pNPP to that of CASP in kidney tissue. Our interpretation would be that the same isozyme is hydrolyzing the S-P bond at one-third the rate that it hydrolyzes the O-P bond.
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PMID:A reinterpretation of phosphatase-N. 629 72


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