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Biomedical subjects

M Yamaguchi

Publications and source records attributed to M Yamaguchi.

At least 883 records · Page 49Linked to original sources
PubMed1994

Serum release of hepatic calcium-binding protein regucalcin by liver injury with galactosamine administration in rats.

Whether calcium-binding protein regucalcin, which mainly localizes in liver, is released into the serum by liver injury was investigated in rats administered galactosamine. Galactosamine (25 mg/100 g body weight) was intraperitoneally administered 3 times at 2 h intervals in rats, and the animals were sacrificed at 10, 24 and 48 h after the first administration of galactosamine. Liver regucalcin mRNA levels were clearly reduced at 24 and 48 h after galactosamine administration with estimating for Northern blotting assay. When hepatic regucalcin concentration was estimated by enzyme-linked immunoadsorbent assay (ELISA) with rabbit-anti-regucalcin IgG, liver regucalcin concentration was not significantly altered by galactosamine administration. Serum regucalcin concentration was markedly elevated at 10 and 24 h after the first administration of galactosamine. Serum transaminases (GOT and GPT) activities were significantly increased by galactosamine administration, indicating that liver injury was induced. The present study demonstrates that liver regucalcin is released into the serum by liver injury with galactosamine administration in rats.

Alanine Transaminase↗
PubMed1994

Synthesis of polymer-coated mixed-functional packing materials for direct analysis of drug-containing serum and plasma by high-performance liquid chromatography.

Silicone polymer-coated mixed-functional (PCMF) silica packing materials were developed for the direct determination of drugs contained in serum or plasma. The new stationary phases were prepared by the following three-step procedure: (1) coating porous silica with a silicone polymer; (2) partial introduction of hydrophobic groups; and (3) introduction of hydrophilic groups. Two PCMFs were synthesized, one having polyoxyethylene groups as hydrophilic groups (PCMF-POE) and the other having oligoglyceryl groups (PCMF-OG). PCMF-POE showed higher recoveries for injected proteins and a greater overall retention for drug molecules than PCMF-OG. PCMF-POE did not show any column deterioration in 500 serum sample injections (10 ml in total).

Carbamazepine↗
PubMed1994

Molecular imaging of Na+,K(+)-ATPase in purified kidney membranes.

Ion channels and pumps in cell membranes consist of multiple transmembrane segments that are thought to be critical for transport of ions. Channel structures constituted by these transmembrane segments are characteristic of ion channels, whereas such structures have not been identified in ion pumps until now. By applying atomic force microscopy on Na+,K(+)-ATPase molecules in canine kidney membranes under tapping mode, we identified a hollow in the protein with a characteristic internal diameter of 6-20 A and an external diameter of 20-55 A depending upon treatment conditions. This hollow may be interpreted as a channel-like conformation of Na+,K(+)-ATPase. In the regions where the proteins were absent, lipid head structures with 2 A width and 6 A length were imaged in an orthorhombic lattice.

Animals↗
PubMed1994

Regulation of ferritin synthesis in macrophages by oxygen and a sulfhydryl-reactive agent.

Ferritin synthesis is known to be regulated translationally by specific mRNA-protein interactions between an iron-responsive element (IRE) localized in the 5' untranslated region of ferritin mRNA and IRE-binding protein (IRE-BP). Binding of IRE-BP to IRE depresses its translation. In the present study, we demonstrated that ferritin synthesis in macrophages is strongly induced under hypoxic conditions by diethylmaleate, a sulfhydryl-reactive agent. The induction by diethylmaleate decreased as the oxygen tension rose. O2- is involved in this oxygen effect, because the induction was prevented when O2(-)-generating agents were present. Actinomycin D did not inhibit the ferritin synthesis induced by diethylmaleate under hypoxi. These results suggest that O2- is involved in post-transcriptional regulation of ferritin synthesis.

Animals↗
PubMed1994

In vitro effects of CINC/gro, a member of the interleukin-8 family, on interleukin-6 secretion by rat posterior pituitary cells.

We investigated the effects of CINC/gro on IL-6 secretion by rat posterior pituitary cells. CINC/gro immunoreactivity was already detected in 1-h conditioned medium of normal posterior pituitary cells, and it increased significantly in a time-dependent manner during the first 24 h of culture. This immunoreactivity could be induced by TNF-alpha in a dose-dependent manner. On the other hand, CINC/gro stimulated IL-6 secretion by posterior pituitary monolayer cultures in a concentration dependent manner. Thus, CINC/gro significantly (P < 0.01) increased the secretion of IL-6 within 13 h of incubation, and this effect continued throughout 24 h of incubation. The stimulatory effect of 100 ng/ml CINC/gro on IL-6 secretion was completely blocked by 24-h incubation with 100 ng/ml IAP. These data demonstrate a new biological activity for CINC/gro in the posterior pituitary system.

Animals↗
PubMed1994

D-type cyclin-binding regions of proliferating cell nuclear antigen.

Proliferating cell nuclear antigen (PCNA) is an auxiliary protein for DNA polymerase delta and is required for both DNA replication and DNA repair. PCNA forms complexes with D-type cyclins, candidate G1 cyclins in mammalian cells. To better understand the functions of the complexes, we examined interactions between PCNA and D-type cyclins, using in vitro-translated mouse PCNA and mouse cyclin D1 or D3 fused to glutathione S-transferase (GST). Analysis of a set of deletion mutants of PCNA revealed that either the N-terminal (residues 2-64) or the C-terminal (residues 197-228) region is necessary for association with D-type cyclins. The cyclin binding of the chimeric protein of the N-terminal (residues 1-68) or the C-terminal (residues 195-261) region of PCNA and rat DNA polymerase beta which does not bind to the cyclins by itself supports this notion. The purified recombinant mouse PCNA expressed in Escherichia coli bound to the D-type cyclin-GST fusion proteins, thereby suggesting that PCNA binds directly to D-type cyclins, without the requirement of other cellular factors. This is apparently the first report on the structure-function relationship of PCNA which may link DNA replication and DNA repair with cell cycle control.

Animals↗
PubMed1994

Determination of urinary glucuronide conjugates by high-performance liquid chromatography with pre-column fluorescence derivatization.

A simple and highly sensitive high-performance liquid chromatographic method for the direct determination of urinary glucuronide conjugates is described. The method is based on the direct derivatization of the glucuronic acid moiety in glucuronide conjugates with 6,7-dimethoxy-1-methyl-2 (1 H)-quinoxalinone-3-propionylcarboxylic acid hydrazide. The derivatization reaction proceeds in aqueous solution in the presence of pyridine and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide at 0-37 degrees C. The resulting fluorescent derivatives are separated on a C18 column using methanol-acetonitrile-0.5% triethylamine in water (1:1:2, v/v) as mobile phase, and are detected spectrofluorimetrically at 445 nm with excitation at 367 nm. The detection limits (signal-to-noise ratio = 3) for the glucuronides are 13-48 fmol for an injection volume of 10 microliters (130-480 fmol per 5 microliters of human urine). The method was applied to the measurement of etiocholanorone-3-glucuronide and androsterone-3-glucuronide in human urine. The method is simple and rapid without conventional liquid-liquid extraction of the glucuronides from urine.

Adult↗
PubMed1994

Differential effect of p53 on the promoters of mouse DNA polymerase beta gene and proliferating-cell-nuclear-antigen gene.

A plasmid carrying the 5' flanking region of the mouse proliferating-cell-nuclear-antigen (PCNA) gene or DNA polymerase beta gene was fused with the chloramphenicol acetyltransferase (CAT) gene, then cotransfected into mouse N18TG2 cells with the expression plasmid for the p53 gene. Expression of the wild-type p53 repressed the CAT expression directed by the PCNA gene promoter, while it had little effect on the DNA polymerase beta gene promoter. RNase protection analysis revealed that the repression of the PCNA gene promoter by p53 was at the transcription step. Analysis with various deletion mutants in the PCNA gene promoter revealed that a specific sequence is not required for the repression, suggesting that p53 represses the PCNA gene promoter by interacting with some components of the basic transcription machinery. By analysis with various deletion mutants in the DNA polymerase beta gene promoter, we identified the unique 10-bp palindromic sequence (-24 to -15), in the presence of which p53 was not able to repress the promoter activity. This sequence conferred resistance to p53 repression onto the PCNA gene promoter, when it was placed 21-bp upstream from the transcription-initiation site.

Animals↗
PubMed1994

Involvement of Ca(2+)-stimulated adenosine 5'-triphosphatase in the Ca2+ releasing mechanism of rat liver nuclei.

The regulatory role of Ca(2+)-stimulated adenosine 5'-triphosphatase (Ca(2+)-ATPase) in Ca2+ transport system of rat liver nuclei was investigated. Ca2+ uptake and release were determined with a Ca2+ electrode. Ca(2+)-ATPase activity was calculated by subtracting Mg(2+)-ATPase activity from (Ca(2+)-Mg2+)-ATPase activity. The release of Ca2+ from the Ca(2+)-loaded nuclei was evoked progressively after Ca2+ uptake with 1.0 mM ATP addition, while it was only slightly in the case of 2.0 mM ATP addition, indicating that the consumption of ATP causes a leak of Ca2+ from the Ca(2+)-loaded nuclei. The presence of N-ethylmaleimide (NEM; 0.1 mM) caused an inhibition of nuclear Ca2+ uptake and induced a promotion of Ca2+ release from the Ca(2+)-loaded nuclei. NEM (0.1 and 0.2 mM) markedly inhibited nuclear Ca(2+)-ATPase activity. This inhibition was completely blocked by the presence of dithiothreitol (DTT; 0.1 and 0.5 mM). Also, DTT inhibited the effect of NEM (0.1 mM) on nuclear Ca2+ uptake and release. Meanwhile, verapamil and diltiazem (10 microM), a blocker of Ca2+ channels, did not prevent the NAD+ (1.0 and 2.0 mM), zinc sulfate (1.0 and 2.5 microM) and arachidonic acid (10 microM)-induced increase in nuclear Ca2+ release, suggesting that Ca2+ channels do not involve on Ca2+ release from the nuclei. These results indicates that an inhibition of nuclear Ca(2+)-ATPase activity causes the decrease in nuclear Ca2+ uptake and the release of Ca2+ from the Ca(2+)-loaded nuclei. The present finding suggests that Ca(2+)-ATPase plays a critical role in the regulatory mechanism of Ca2+ uptake and release in rat liver nuclei.

Adenosine Triphosphate↗
PubMed1994

Hepatic calcium-binding protein regucalcin in released into the serum of rats administered orally carbon tetrachloride.

The change in calcium-binding protein regucalcin, mainly localized in liver, in the liver and serum of rats received a single oral administration of carbon tetrachloride (50%; 1.0 ml/100 g body weight) was investigated. The change of regucalcin mRNA levels in the liver was analyzed by Northern blotting using liver regucalcin cDNA (0.6 kb). At 10 and 24 h after the administration, liver regucalcin mRNA levels were reduced markedly. Moreover, regucalcin concentrations in the liver and serum was estimated by enzyme-linked immunoadsorbent assay (ELISA) with rabbit-anti-regucalcin IgG. Administration of carbon tetrachloride (CCl4) induced a significant decrease in liver regucalcin concentration and a corresponding elevation of serum regucalcin concentration at 24 h after the administration. An appreciable increase in serum regucalcin concentration was seen at 2 h after the administration. Meanwhile, serum transaminases (GOT and GPT) activities were significantly increased by CCl4 administration, indicating that liver injury is induced. The present study demonstrates that hepatic regucalcin is released into the serum of rats administered orally CCl4, suggesting that the estimation of serum regucalcin is a useful tool for diagnosis of liver injury.

Administration, Oral↗
PubMed1994

Effect of beta-alanyl-L-histidinato zinc on differentiation of osteoblastic MC3T3-E1 cells: increases in alkaline phosphatase activity and protein concentration.

The effect of beta-alanyl-L-histidinato zinc (AHZ) on bone cell function was investigated in osteoblastic MC3T3-E1 cells. Cells were cultured for 3 days at 37 degrees C in a CO2 incubator in plastic dishes containing alpha-modified minimum essential medium supplemented with 10% fetal bovine serum. After the cultures, the medium was exchanged for that containing 0.1% bovine serum albumin plus AHZ (10(-7)-10(-5) M) or other reagents, and the cells were cultured further for appropriate periods of time. The presence of AHZ (10(-7)-10(-5) M) produced a remarkable increase of alkaline phosphatase activity and protein concentration in osteoblastic cells. Thus increase were seen with the prolonged cultivation (12-21 days). With the culture of 1, 3 and 12 days, the effect of AHZ (10(-6) M) to increase alkaline phosphatase activity and protein concentration was more intensive than the effect of zinc sulfate (10(-6) M). The AHZ effects were completely abolished by the presence of cycloheximide (10(-6) M), indicating that AHZ stimulates protein synthesis in the cells. The present study suggests that AHZ has a stimulatory effect on cell differentiation, and that this effect is partly involved on protein synthesis in osteoblastic cells.

Alkaline Phosphatase↗
PubMed1994

Repression of regulatory factor for Drosophila DNA replication-related gene promoters by zerknüllt homeodomain protein.

The expression of the chloramphenicol acetyltransferase gene under control of the 1152-base pair 5'-flanking region (-1107 to +45 nucleotide positions with respect to the major transcription initiation site) of the Drosophila DNA polymerase alpha gene was repressed by cotransfection into Drosophila Kc cells with a zerknüllt (zen)-expressing plasmid as previously observed with the proliferating cell nuclear antigen (PCNA) gene promoter. The expression of the zen resulted in reduction of the abundance of mRNA for the transfected chloramphenicol acetyltransferase gene and also mRNAs for both DNA polymerase alpha and PCNA. Results obtained using various deletion derivatives of the promoter region and chemically synthesized oligonucleotides of the DNA replication-related element (DRE), a positive cis-acting element found in both DNA polymerase alpha and PCNA genes, revealed that the DRE sequences are responsible to repression by Zen protein. The nuclear extract of Kc cells transfected by the zen-expressing plasmid contained lesser amounts of the DRE-binding factor (DREF) than that of untransfected or mutant zen-transfected cells. These results suggest that the Zen protein represses expression of DNA replication-related genes by reducing DREF, although the detailed mechanism of the repression remains to be elucidated.

Animals↗
PubMed1994

Effect of phorbol 12-myristate 13-acetate on Ca2+-ATPase activity in rat liver nuclei.

The effect of phorbol 12-myristate 13-acetate (PMA) on Ca(2+)-ATPase activity in rat liver nuclei was investigated. Ca(2+)-ATPase activity was calculated by subtracting Mg(2+)-ATPase activity from (Ca(2+)-Mg(2+)-ATPase activity. The nuclear Ca(2+)-ATPase activity was significantly increased by the presence of PMA (2-20 microM) in the enzyme reaction mixture; the maximum effect was seen at 10 microM. The PMA (10 microM)-increased Ca(2+)-ATPase activity was not blocked by the presence of staurosporine (2 microM) or dibucaine (2 and 10 microM), an inhibitor of protein kinase. Meanwhile, vanadate (20 and 100 microM) caused a significant reduction in the nuclear Ca(2+)-ATPase activity increased by PMA (10 microM). The present finding suggests that PMA has an activating effect on liver nuclear Ca(2+)-ATPase independent of protein kinase.

Adenosine Triphosphate↗
PubMed1994

Enhancement of differentiation induction of mouse myelomonocytic leukemic cells by extracellular ATP.

We examined the effect of ATP on the terminal differentiation of mouse myelomonocytic leukemic M1 cells to macrophages. Although ATP alone did not induce M1 cell differentiation, addition of ATP with the differentiation inducer, interleukin 6 (IL-6), enhanced the induction of differentiation by IL-6 about two-fold. Comparison among several adenine nucleotides revealed that the order of effectiveness on differentiation enhancement was ATP > ADP > AMP > or = adenosine. Using reactive blue 2, a P2 receptor antagonist, we confirmed that the effect of ATP on the stimulation of differentiation was mediated through the P2 purinergic receptor. Examination of cytosolic [Ca2+] elevation by ATP and comparison of potency of differentiation enhancement among several ATP analogs demonstrated that the effect of differentiation enhancement was mediated through P2y purinergic receptors expressed on M1 cell surface. Within 3 h of exposure, ATP alone slightly increased the expression of differentiation-related immediate early response genes, c-myc and JunB, and ATP also enhanced the IL-6-induced expression of these genes. Induction of JunB expression by ATP analogs correlated with their potency of differentiation enhancement, which suggested that induction of JunB by ATP is one of signaling pathways involved in the exertion of its differentiation-enhancing effect.

1-Methyl-3-isobutylxanthine↗
PubMed1994

The effect of leukotriene C4 on the permeability of brain capillary endothelial cell monolayer.

The role of leukotrienes as mediator of brain edema is still controversial. Recently, the ability of gamma-GTP to act as enzymatic barrier and to inactivate leukotrienes in normal brain capillaries was pointed out. A hypothesis tested in our experiments was that Leukotriene C4 (LTC4) increases permeability of a cerebral capillary endothelial monolayer which lacks gamma-GTP activity. Brain capillary endothelial cells were obtained of 10 rats from cerebral cortex by an enzymatic isolation procedure. The cells have an intact function, however, lack gamma-GTP activity. The endothelial cells were cultured on an optically clear collagen membrane mounted on a plastic frame. Effects of bradykinin (1 x 10(-5) M) and LTC4 (1 x 10(-7) M, 1 x 10(-6) M, 5 x 10(-6) M, 1 x 10(-5) M) were tested on permeability of the endothelial cell monolayer by measuring leakage of 14C-sucrose. The effect of LTC4 and bradykinin on intracellular calcium was studied by laser scanning confocal microscopy. LTC4 did not increase permeability of the brain capillary endothelial cell monolayer which lacked gamma-GTP activity. LTC4 did neither increase the concentration of intracellular calcium. Differences of LTC4 receptor function in normal brain capillaries and tumor capillaries remain to be studied.

Animals↗
PubMed1994

Cerebral blood flow of rats with water-intoxicated brain edema.

Water intoxication brain edema was produced in rats by intraperitoneal loading of excessive amounts of distilled water (DW). In 10% and 20% groups, DW in amounts of 10 or 20% of body weight was injected, respectively. Water content of brain tissue increased proportionally to the amount of injected water, as follows: 79.8% of wet weight in control, 80.5 and 82.4 in 10% and 20% DW groups, respectively. Since cerebral blood flow (CBF) values measured by laser Doppler (LD) flowmetry were found to give a good correlation with those by hydrogen clearance method in a preliminary experiment, CBF measurement were carried out using LD flowmetry thereafter. Before the injection, CBF values were around 50 ml/min/100 g. Two hours after the water loading, CBF values in 10% and 20% DW groups were 25.6 and 20.3 ml/100 g/min, respectively. CBF values under these edematous condition decreased significantly (p < 0.001 by paired t-test) in proportion to the severity of the brain edema.

Animals↗