Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.2.1.23 (beta-galactosidase)
 14,648 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have developed an enzyme linked immunosorbent assay (ELISA) for antimitochondrial antibody. Polyvinyl microtiter plate wells are coated with partially purified rat kidney mitochondria, and excess protein binding sites are blocked with bovine serum albumin. Human serum, diluted 1:1,000, is incubated for 1 hr. Then beta-galactosidase-goat-anti-human IgG (H + L) is added followed by the substrate, p-nitrophenyl-beta-D-galactopyranoside. The plates are then read at 404 nM in a microelisa autoreader. A positive result was defined as optical density greater than or equal to 0.100, more than 5 standard deviations above the mean of 36 normal individuals. With this technique, 56 of 60 patients with primary biliary cirrhosis were positive for antimitochondrial antibody (93%), mean O.D., 0.456 +/- 0.031 S.E. Seventeen of 17 patients with extrahepatic bile duct obstruction and 14 or 14 patients with alcoholic cirrhosis were negative. Only 1 of 29 patients with chronic active liver disease was positive (4%). Antinuclear antibody and antimicrosomal antibody do not bind in this assay, and activity is absorbed from sera by preincubation with suspensions of rat kidney mitochondria. The ELISA is approximately 20 times more sensitive than a quantitative microtiter complement fixation technique and more convenient than radioimmunoassay. It is rapid, quantitative and uses stable reagents. In contrast to immunofluorescence techniques, it is not affected by observer interpretation.
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PMID:An enzyme-linked immunosorbent assay (ELISA) for detecting antimitochondrial antibody. 637 51

Although the cellular mechanisms controlling bile flow and biliary lipid secretion are unclear, morphologic data suggest that intracellular vesicles may be involved. Therefore, to investigate the role of hepatocyte lysosomes in bile flow and biliary lipid secretion, we studied the effect of cholestasis on biliary lipid output and on lysosomal enzyme activities and total protein concentration in liver and bile. Castrated male rats were treated with ethinyl estradiol at 0.5 or 5 mg/kg per day for 5 days; bile was collected through a complete bile fistula hourly for 4 hours, and then liver homogenates were prepared. Bile acids, cholesterol, and phospholipid were measured in bile, and three lysosomal glycosidases (beta-glucuronidase, beta-galactosidase, and N-acetyl-beta-glucosaminidase) and total protein were measured in bile and liver. Ethinyl estradiol inhibited bile flow in a dose-dependent fashion; it also inhibited bile acid and phospholipid outputs. In contrast, a marked and parallel increase in the biliary outputs of all three lysosomal hydrolases was observed after high-dose ethinyl estradiol; no change in the biliary concentration of total protein was found. Our data suggest that bile flow and biliary lipid secretion involve cellular mechanisms other than vesicular transport by lysosomes.
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PMID:Dissociation of bile flow and biliary lipid secretion from biliary lysosomal enzyme output in experimental cholestasis. 678 57

Lysosomal enzyme activity in the bile and blood serum was compared in mice with experimental intrahepatic cholestasis induced by alpha-naphthyl isothiocyanate and Triton WR 1339. Triton WR 1339 increases the synthesis of cholesterol (fatty acid precursor) in liver cells. The development of intrahepatic cholestasis was confirmed by the increase in activities of alkaline phosphatase and gamma-glutamyltransferase in blood serum. Administration of Triton WR 1339 in a dose of 100 mg/100 g was followed by a 10-fold increase in beta-galactosidase activity (hepatocyte lysosomal enzyme) in the bile, but not in the serum of mice. beta-Galactosidase activity significantly increased in the bile, but decreased in the serum of mice after treatment with a-naphthyl isothiocyanate in a dose of 200 mg/kg. Our results indicate that intrahepatic cholestasis is manifested in increased secretion of lysosomal glycosidases into the bile. Bile components can aggravate damage to liver cells by affecting the processes of hepatocyte apoptosis and necrosis.
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PMID:Activity of lysosomal enzymes in the bile and serum of mice with intrahepatic cholestasis. 1914 81