Catalytic site of human protein-glucosylgalactosylhydroxylysine glucosidase: Three crucial carboxyl residues were determined by cloning and site-directed mutagenesis.

Protein-glucosylgalactosylhydroxylysine glucosidase (PGGHG; EC3.2.1.

Enzyme cytochemical study of alkaline phosphatase in rat hepatocytes--change of location after colchicine administration or bile duct ligation.

Changes in location of alkaline phosphatase (ALP) in rat hepatocytes after two different treatments were examined by the enzyme cytochemical technique. Although reaction deposits which indicate ALP activity were observed mainly in the bile canalicular membranes until 6 hr after colchicine administration, they were present not only in the bile canalicular membranes but also in the sinusoidal and lateral membranes 12 hr after drug administration. Eighteen and 24 hr after colchicine administration, a large number of reaction deposits were seen in the sinusoidal membranes and in the mutual membranes between adjacent hepatocytes, where the bile canaliculi were completely deformed and complex interdigitations appeared.

Effect of beta-carotene, sodium ascorbate and cellulose on 1,2-dimethylhydrazine-induced intestinal carcinogenesis in rats.

Male Sprague-Dawley rats were fed a diet containing either 0.005% beta-carotene, 0.02% sodium ascorbate or 1.

Purification and characterization of a 100 kDa glycyrrhizin (GL)-binding protein (gp100) as an effective phosphate acceptor for CK-II and the effect of GL on the phosphorylation of gp100 by CK-II in vitro.

By means of successive GL-affinity and Mono S column chromatographies (HPLC), a 100 kDa GL-binding protein (gp100) was purified from the partially purified CK-II fraction of EAT cells as an effective phosphate acceptor for CK-II. It was found that (i) gp100 (pI 9.0) is copurified with CK-II, Hsp-90 and p34 or p70; (ii) gp100 cross-reacts with anti-human GR; (iii) phosphorylation of gp100 by CK-II is significantly stimulated by 1 microM GL or 0.

The biological significance of glycyrrhizin- and glycyrrhetinic acid derivative-induced selective phosphorylation of histones H2A and H2B by A-kinase in vitro.

Histones H2A and H2B were found to be glycyrrhizin (GL)-binding proteins, because (i) the two histones H2A-H2B pairs were isolated selectively from the crude histone preparations of calf thymus by means of GL-affinity column chromatography (HPLC); (ii) phosphorylation of these two histones by A-kinase was remarkably stimulated by native GL or oGA (a derivative of glycyrrhetinic acid) at 20 microM; and (iii) in the crude histone preparations of calf thymus, these two histones were selectively phosphorylated by A-kinase in the presence of both dsDNA and 20 microM oGA or 20 microM GL. The provided data suggest that the GL-induced selective phosphorylation of histones H2A and H2B by A-kinase may be implicated in the transcriptional activation involved in the biological activities of the drug.

Immunohistochemical detection of alkaline phosphatase in formalin-fixed and paraffin-embedded rat organs by means of avidin-biotin peroxidase complex method.

The author attempted the detection of alkaline phosphatase (ALP) protein in various rat organs, which were fixed with formalin and embedded in paraffin, by means of the avidin-biotin peroxidase complex (ABC) method. Reaction products were detected after a usual incubation time of 30 min at room temperature in anti-ALP antibody. Positive reactions were observed in the kidney and liver but not in the small intestine, and this indicates that the antibody used in the present study reacts only with the tissue-nonspecific type of ALP.

Immunohistochemical detection of alkaline phosphatase in formalin-fixed and paraffin-embedded rat liver.

Alkaline phosphatase (ALP) in rat liver was detected by means of immunohistochemical techniques in ordinary histologic specimens which were fixed with formalin and embedded in paraffin. The specimens were incubated in anti-ALP antibody at room temperature for a longer time, 3hr, than ordinary incubation time. Antigen detection sensitivity and contrast of reaction product of the immunogold silver staining were superior to those of the peroxidase-labeled antibody method.

[The relationship between serum leukotriene B4 and smoking].

Leukotriene B4 (LTB4), an arachidonic acid metabolite released by neutrophils, is involved in the regulation of the host immune response to antigenic stimulation. Furthermore, LTB4 affects the chemokinesis, aggregation, and enzyme release of neutrophils and stimulates activity of cytotoxic T cells, natural killer cells and suppressor T cells. On the other hand, smoking typically results in inflammatory stimulation of the lung, and long-term smoking can cause chronic stimulation.

Effects of sodium and chloride ions on blood pressure in deoxycorticosterone acetate-treated rats.

The effects of sodium (Na+) and chloride ions (Cl-) on blood pressure were studied in rats treated with deoxycorticosterone acetate (DOCA). Four groups were prepared, each consisting of male Wistar rats that underwent heminephrectomy and administration of DOCA: the control group was maintained with tap water, the NaCl group with tap water containing 1% sodium chloride, the NaCit group with tap water containing 1.67% sodium citrate (including an equivalent dose of Na+ to 1% NaCl), and the ChoCl group with tap water containing 1.

Effects of beta-blocking agents on the release of various enzymes in muscular tissues.

We examined the kinetics study of serum enzyme after the administration of beta-blocking agents or alpha-stimulator in the experimental rats. Following the administration of beta-blocking agents, propranolol and pindolol, the serum levels of adenylate kinase, aldolase, lactate dehydrogenase and aspartate aminotransferase as well as that of creatine kinase increased in rats. The same was observed following the administration of noradrenaline (an alpha-stimulator).

Characterization of casein kinase II, and of p98 as one of its effective phosphate acceptors in sea urchin eggs.

CK-II has been partially purified from a 1.5 M KCl extract of unfertilized sea urchin eggs by means of DEAE-cellulose column chromatography, gel filtration on Sephacryl S300, and heparin-agarose column chromatography, successively. This partially purified CK-II was associated with a 98 kDa polypeptide (designated as p98), which was then separated from the kinase by Mono Q column chromatography (HPLC).

Suppression of 125I-uptake in mouse thyroid by seaweed feeding: possible preventative effect of dietary seaweed on internal radiation injury of the thyroid by radioactive iodine.

We conducted an animal experiment to determine how dietary seaweeds rich in iodine and dietary fibers suppress radioactive iodine uptake by the thyroid, using mice and four kinds of experimental diets, three with 1% or 2% powdered fronds of the kelp Laminaria religiosa and 2% powdered laver Porphyra yezoensis, and one with cellulose. Iodine content of a hot-water extract of the kelp was 0.530 +/- 0.

The stimulatory and inhibitory effects of glycyrrhizin and a glycyrrhetinic acid derivative on phosphorylation of lipocortin I by A-kinase in vitro.

The effects of GL and oGA on the activity (phosphorylation of lipocortin I) by A-kinase were investigated in vitro. It was found that (i) phosphorylation of the 35-36 kDa polypeptide by A-kinase was inhibited selectively when the partially purified kinase fraction from EAT cells was incubated with [gamma-32P]ATP in the presence of 60 microM oGA; (ii) the marked polypeptide was identified as a lipocortin I; and (iii) the activity of the kinase was stimulated about 4-fold by 5 microM oGA, but it was inhibited at doses above 25 microM. The drug-induced inhibition of phosphorylation of lipocortin I by A-kinase may be implicated in the anti-inflammatory effect of the drugs.

[Proposal for the nosocomial infection control of methicillin-resistant Staphylococcus aureus (MRSA)].

To determine the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in hospital environments, quantitative isolation of MRSA from environmental and human specimens was performed. It was found that as many as 6 x 10(4) colony forming units (CFU)/100cm2 of MRSA and also methicillin-resistant coagulase-negative Staphylococcus (MRCNS) were isolated from 1) 25-100% of the bed-mattresses tested in 10 out of 11 (90%) hospitals, and also 2) bed sheets for patients, floors of patient wards, laundry, bath-room, toilet and laundry storage-room. And seven and eight palms of 20 patients were contaminated with MRCNS and MRSA, respectively, and MRCNS contamination was revealed in six of medical staffs.

[Newly developed MRSA medium].

We have developed a new selective medium, tentatively named MR(SA)2, for rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) from clinical specimens. MR(SA)2 medium contained modified Müller-Hinton agar supplemented with 75 g of NaCl, 10 g of mannit, 20 mg of bromcresol purple, 20 g of egg-yolk, 4 mg of oxacillin, and 12.5 mg of ceftisoxime per 1,000 ml.

Isopropylidine maltoheptosyl fructofuranoside, doubly blocked substrate for determination of endoamylase activity.

We synthesized o-(4,6-o-isopropylidene-alpha-D-glucopyranosyl)-(1----4)- [o-alpha-D-glucopyranosyl-(1----4])5-o-alpha-D-glucopyranosyl-(1----2)- alpha-D-fructofuranoside (IPG7F) and developed an assay for determining the activity of amylase in human serum and urine by using this substrate. Glucoamylase, alpha-glucosidase, and mannitol dehydrogenase are used as coupling enzymes. The coupled reactions are monitored by continuously measuring the oxidation rate of NADH.

Effect of dietary kelp on lipid peroxidation and glutathione peroxidase activity in livers of rats given breast carcinogen DMBA.

In our previous study of female rats, a diet containing 2% of powdered fronds of Laminaria religiosa (kelp) had a remarkable inhibitory effect on mammary tumorigenesis induced by orally given 7,12-dimethylbenz[a]anthracene (DMBA). In this study, the effect of the dietary kelp on lipid peroxidation and glutathione peroxidase (GSH-Px) activity in the liver was studied in the same experimental system. Two separate short- and long-term experiments were carried out on rats fed a diet with 2% kelp.

Human monocyte antigens in a random sample from Japan.

We studied a random sample of Japanese and compared both Japanese and German monocyte antigen frequencies and performed a segregation analysis in Japanese families. In Japanese, the following gene frequencies were established: 0.1033 for HMA-A1; 0.

Histochemical and immunohistochemical study on the expression of alkaline phosphatase, gamma-glutamyl transpeptidase and alpha-fetoprotein during the process of rat hepatocyte proliferation.

The changes in the activity and the localization of alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (gamma-GTP) and alpha-fetoprotein (AFP) were examined during cell regeneration in the galactosamine-injured rat liver. D-galactosamine was injected i.p.

Characterization of a basic polypeptide-activated protein kinase in the eggs of the silkworm, Bombyx mori.

A basic polypeptide-activated protein kinase (designated BA-kinase), which requires Mn2+ rather than Mg2+ for its activity, has been highly purified from a 1.5 M KCl extract of fertilized and unfertilized eggs of the silkworm, Bombyx mori. It was found that (i) the molecular weight of the purified protein kinase was approx.

Effects of alcohol consumption on mitochondrial aldehyde dehydrogenase isoenzymes in rat liver.

The effects of long-term and short-term exposure of rats to ethanol on aldehyde dehydrogenase (ALDH) activity in the liver mitochondria were investigated. The specific activities of mitochondrial high Km ALDH and low Km ALDH after the prolonged administration of ethanol were both increased to levels about 2.5 times that of the control group.

Inhibitory effect of glycyrrhizin on polypeptide phosphorylation by polypeptide-dependent protein kinase (kinase P) in vitro.

The anti-viral mechanism of glycyrrhizin (GL) has been investigated by considering in vitro effects on polypeptide phosphorylation. It was found that GL (i), at low doses, selectively inhibits protein phosphorylation by Kinase P, but has not significant effects on the activities of other kinases (Kinase A, Kinase C and histone kinase); (ii) binds directly to Kinase P and reduces kinase activity in a dose-dependent manner; and (iii) inhibits vesicular stomatitis virus (VSV)-associated kinase activity. These observations strongly suggest that direct binding of GL to the virus causes the direct inactivation of virus-associated kinase and the reduction of the viral infectivity.