Inhibition of monoamine oxidase by 3,4-dihydroxyphenylserine.

The effects of diastereomers of 3,4-dihydroxyphenylserine (DOPS) on the enzyme activity of monoamine oxidase (MAO) in human placenta and liver mitochondria were examined. Both L- and D-threo-DOPS were found to inhibit MAO-A in human placental mitochondria in competition with the substrate, and the Ki values for L- and D-threo-DOPS obtained were 68.3 and 125 microM, respectively.

Specific binding of glycosylated beta-galactosidase to bovine brain synaptic membrane.

The binding of a series of glycosylated beta-galactosidases to a fraction rich in synaptic membrane of bovine brain was examined. beta-galactosidase modified with p-aminophenyl beta-D-galactopyranoside (beta-D-Gal beta-gal) was found the most effective in binding to synaptic membrane, followed by that modified with beta-D-glucopyranoside, whereas the enzyme modified with p-aminophenyl derivatives of alpha-D-galactopyranoside, alpha-D-glucopyranoside, and alpha- and beta-L-fucopyranoside were found not to bind to the membrane. The binding was dependent on time, temperature, and pH; the maximal binding was obtained within 15 min at 4 degrees C and the optimal pH was approximately 4.

A family with beta-galactosidase deficiency: three adults with atypical clinical patterns.

Three adult patients in a single family showed severe myoclonus, ataxia, and pyramidal signs. Enzymatic analysis of lymphocytes, plasma, and cultured skin fibroblasts showed marked deficiency of beta-galactosidase activity, more profound with GM1 ganglioside than with another natural substrate, asialofetuin. Other lysosomal hydrolases were normal.

A microassay for acid beta-galactosidase activity toward asialofetuin.

To study the enzymatic properties of beta-galactosidase from the patients with a beta-galactosidase deficiency such as GM1 gangliosidosis, determination of enzymatic activity with naturally occurring substrates, asialofetuin in addition to another natural substrate, GM1 ganglioside, is essentially required. With a previously reported, simple and sensitive fluorometric assay for GM1 ganglioside beta-galactosidase using high performance liquid chromatography (HPLC), optimal reaction conditions were determined for the assay of acid beta-galactosidase activity toward asialofetuin in skin fibroblast homogenates. Under these conditions, reduced enzymatic activities could be detected in cultured skin fibroblasts from patients with type 1 and 3 GM1 gangliosidoses and mucopolysaccharidosis IV-B (Morquio B syndrome).

Assay of glucocerebrosidase using a fluorescent analogue of glucocerebroside for the diagnosis of Gaucher disease.

For the diagnosis of homozygotes and heterozygotes of Gaucher disease, glucocerebrosidase (glucocerebroside beta-D-glucoside glucohydrolase, EC 3.2.1.

Laparoscopic-assisted serial biopsy of the bovine kidney.

A serial renal biopsy with endoscopy was done, using 10 calves and a cow. In 8 of the calves, biopsy materials were collected 3 to 5 times over a period of 15 to 41 days. A laparoscope was inserted into the peritoneal cavity from the center of the right paralumbar fossa through the outer cannula of the trocar.

[Specific in vitro effects of methotrexate entrapped into liposomes bearing antibody fragments against human chorionic gonadotropin on cell growth of cultured human choriocarcinoma cells].

Methotrexate (MTX) was entrapped into liposomes, to which rabbit antibody fragments (Fab') against human chorionic gonadotropin (hCG) had been coupled covalently, using N-[4-(p-maleimidophenyl) butyryl] phosphatidyl ethanolamine. By this system, The resulting molar ratio in the prepared liposomes was 15 MTX per Fab', and the activities of both MTX and the antibody were well preserved. The inhibitory effects of MTX entrapped in liposomes bearing anti-hCG Fab' (Lip[MTX, anti-hCG]) on the growth of choriocarcinoma cells (BeWo) in vitro were examined.

[Distribution of an antibiotic (cefotiam) to the breast tissue].

In order to demonstrate the distribution of an antibiotic to normal breast tissues and its penetration into the axillary wound exudate, 1 g of a new cephem antibiotic, cefotiam (CTM), was infused intravenously over 1 hour in 14 patients with breast cancer before and after mastectomy. CTM concentrations were assayed by the cylinder-plate method, using Proteus mirabilis ATCC 21100 as the test organism. Overall CTM levels in the normal breast tissues ranged from 3.

Effect of glycolipids contained in liposomes on the incorporation of beta-galactosidase into specific organs.

A series of glycolipids were examined to find a system capable of targeting liposomes into specific organs of rats. Sulfatide was found to be the best among the components of liposomes examined for delivering the entrapped enzyme, beta-galactosidase from Aspergillus oryzae, into the brain and liver; gangliosides, for the spleen; and sphingomyelin, for the lung. To introduce the enzyme into the liver, galactocerebroside was far better than glucocerebroside.

Application of a GM1 ganglioside beta-galactosidase microassay method to diagnosis of GM1 gangliosidosis.

The enzymatic diagnosis of GM1 gangliosidosis, including the diagnosis of heterozygosity, requires a microassay of GM1 ganglioside beta-galactosidase activity in lymphocytes and cultured skin fibroblasts. We have adopted high-performance liquid chromatography (HPLC) to the assay of this enzyme and can measure the activity in crude samples fluorometrically. Reaction conditions were examined to determine those optimal for the assay of GM1 ganglioside beta-galactosidase activity in lymphocyte and skin fibroblast homogenates.

A fluorometric microassay procedure for monitoring the enzymatic activity of GM1-ganglioside beta-galactosidase by use of high-performance liquid chromatography.

For the measurement of the enzymatic activity of GM1-ganglioside (II3 NeuAcGgOse4Cer, galactosyl-N-acetylgalactosaminyl-(N-acetylneuraminosyl) galactosyl-glucosylceramide) beta-galactosidase in crude enzyme samples, a microassay using nonradioisotopic GM1-ganglioside was devised. To reduce the volume of the reaction mixture and eliminate the interferences due to the fluorescent contaminants in the reaction mixture, NADH, a product after the oxidation of the released galactose with NAD and beta-galactose dehydrogenase, was fluorometrically estimated by use of high-performance liquid chromatography. By this method, as little as 10 pmol of galactose can be detected.

Alteration of the substrate specificity of Aspergillus oryzae beta-galactosidase by modification with polyethylene glycol.

beta-Galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.

Use of a fluorescent analogue of galactocerebroside for assay of galactocerebroside beta-galactosidase activity in skin fibroblasts from patients with Krabbe's disease.

In order to diagnose Krabbe's disease (globoid cell leukodystrophy) or its carrier state in patients, galactocerebroside beta-galactosidase (galactosylceramidase) activity in cultured skin fibroblasts was measured with high-performance liquid chromatography using a fluorescent analogue of galactocerebroside, 1-O-galactosyl-2-N-(1-dimethylaminonaphthalene-5-sulfonyl)-sphingosine, as substrate. The enzyme activities in skin fibroblasts of the patients were found to be dramatically reduced when compared with those of controls. The assay can be adopted for diagnostic use in place of assays using radioisotope-labeled natural substrate.

Immobilized D-amino acid oxidase.

1. D-Amino acid oxidase (D-amino acid: oxygen oxidoreductase (deaminating), EC 1.4.

Permeability of amino acids into liposomes.

1. A simple and rapid assay for the measurement of permeability of amino acids into liposome membrane was carried out by using the liposomes trapping D-amino acid oxidase (D-amino acid: O2 oxidoreductase (deaminating), EC 1.4.

Oxidation of sarcosine and N-alkyl derivatives of glycine by D-amino-acid oxidase.

1. Sarcosine was oxidized by D-amino-acid oxidase (D-amino-acid: O2 oxidoreductase (deaminating), EC 1.4.