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Y Sakamoto

Publications and source records attributed to Y Sakamoto.

At least 199 records · Page 11Linked to original sources
PubMed1998

Troglitazone reduces free fatty acid-induced insulin resistance in perfused rat hindquarter.

In order to study the effects of troglitazone on insulin resistance associated with elevated plasma free fatty acid (FFA), the hindquarters of rats treated with troglitazone for 14 days were perfused with a medium containing 15 mmol/l glucose, 0-1,000 microU/ml insulin, and 0 or 1.0 mmol/l palmitate. In the absence of palmitate, net glucose uptake was similar between control and troglitazone-treated rat hindquarters at each insulin concentration: 47 +/- 10 and 45 +/- 9 mumol for 30 min at 0 microU/ml insulin; and 94 +/- 14 and 99 +/- 13 mumol for 30 min at 1,000 microU/ml insulin respectively. Addition of palmitate abolished insulin stimulation of net glucose uptake in control rat hindquarter but did not decrease it in troglitazone-treated rat hindquarter (60 +/- 11 vs 99 +/- 12 mumol for 30 min at 1,000 microU/ml insulin, p < 0.01). It is suggested that the "FFA-tolerant effect" is one mechanism by which troglitazone reduces insulin resistance.

Animals↗
PubMed1997

Cytokine-induced neutrophil chemoattractant gene expression in the rat hypothalamus by osmotic stimulation.

Cytokine-induced neutrophil chemoattractant (CINC) is one of the chemokines and has chemotaxity for neutrophils. Recently, we found the presence of stress-sensitive CINC expression in the hypothalamic nuclei such as the paraventricular nucleus. Since CINC was predominantly co-localized with vasopressin in the supraoptic nucleus (SON), we investigated the effect of hyperosmotic challenge on CINC mRNA in the hypothalamus. We found that CINC mRNA expression in the hypothalamus was augmented within 30 min following osmotic stimulation and immediately returned to the basal level. The suckling, which is a stimulation to oxytocin neurons in the SON, has no effect on CINC mRNA expression in the hypothalamus. This is the first evidence that the chemokine in the brain is activated by osmotic stimulation.

Animals↗
PubMed1997

Ursodeoxycholic acid enhances glucocorticoid-induced tyrosine aminotransferase-gene expression in cultured rat hepatocytes.

Ursodeoxycholic acid (UDCA) is an effective treatment for immune-mediated liver diseases, suggesting that UDCA is functionally similar to glucocorticoids (GCs). We investigated the effects of UDCA on the enzyme activity and the mRNA levels of tyrosine aminotransferase (TAT), a hepatocyte-specific marker of GC action, in primary cultured rat hepatocytes. Addition of UDCA resulted in a significant increase in TAT activity in the presence of dexamethasone (DEX), compared with DEX alone, and this increase was completely suppressed by sphingosine, a protein kinase C (PKC) inhibitor, or actinomycin D, a transcriptional inhibitor. UDCA could not induce TAT activity in the absence of DEX. UDCA increased the TAT mRNA levels in the presence of DEX. In conclusion, UDCA enhances the GC-induced TAT-gene expression in hepatocytes, and UDCA-activated PKC may play a role in this upregulation.

Animals↗
PubMed1997

Inhibition of the DNA-binding and transcriptional repression activity of the Wilms' tumor gene product, WT1, by cAMP-dependent protein kinase-mediated phosphorylation of Ser-365 and Ser-393 in the zinc finger domain.

The Wilms' tumor suppressor gene, WT1, encodes a transcription factor in the zinc finger family, which binds to GC-rich sequences and functions as a transcriptional activator or repressor. The WT1 protein plays a crucial role in urogenital development in mammals and its function is thought to be conserved during vertebrate evolution. Although accumulating evidence suggests that WT1 regulates a subset of genes including growth factor and growth factor receptor genes, little is known about regulators or signal cascades that could modulate the function of WT1. In this study, we show that the WT1 protein expressed exogenously in fibroblasts was phosphorylated in vivo, and that treatment with forskolin, which activates the cAMP-dependent protein kinase (PKA) in vivo, induced phosphorylation of additional sites in WT1. We identified the forskolin-induced phosphorylation sites as Ser-365 and Ser-393, which lie in the zinc finger domain in zinc fingers 2 and 3, respectively. PKA phosphorylated WT1 at Ser-365 and Ser-393 in vitro, as well as at additional sites, and this phosphorylation abolished the DNA-binding activity of WT1 in vitro. Using WT1 mutants in which Ser-365 and Ser-393 were mutated to Ala individually and in combination, we showed that phosphorylation of these sites was critical for inhibition of DNA binding in vivo. Thus, coexpression of the PKA catalytic subunit with wild type WT1 reduced the level of WT1 DNA-binding activity detected in nuclear extracts, and decreased transcriptional repression activity in vivo. In contrast to wild type WT1, all of the phosphorylation site mutants retained significant DNA-binding activity and repression activity in the presence of PKA. Analysis of the mutants showed that phosphorylation of Ser-365 and Ser-395 had additive inhibitory effects on WT1 DNA-binding in vivo and that phosphorylation at both sites was required for neutralization of repression activity. Therefore, we conclude that PKA modulates the activity of WT1 in vivo through phosphorylation of Ser-365 and Ser-393, which inhibits DNA binding. This in turn results in a decrease in WT1 transcriptional repression. Our findings provide the first evidence that the function of WT1 can be modulated by its phosphorylation in vivo.

Animals↗
PubMed1997

A simple method to confirm patency of the graft bile duct during living-related partial liver transplantation.

A bile duct stricture caused by an inadvertent ligation or suture during donor operation is rare, but not a negligible complication after living-related liver transplantation (LRLT). We present here a simple technique to prevent this kind of complication. Before bilioenteric anastomosis, a rod-shaped surgical probe was introduced into the bile ducts of the graft to examine their patency. The position of the surgical probe and the liver segment in which the probe proceeded was checked using ultrasonography. Using this technique in all of our 60 cases of LRLT, we have had no experience of this bile duct complication. We recommend this technique to be adopted in LRLT and split-liver transplantation to diminish the risk of biliary complications.

Adult↗
PubMed1997

Human alveolar macrophages synthesize endothelins by thrombin.

Alveolar macrophages (AM) play pivotal roles in the defense mechanism and the regulation of inflammatory processes in the airways. Macrophages express receptors for thrombin on their surfaces, and thrombin induces the chemotaxis and the proliferation of macrophages. Thrombin acts on vascular endothelial cells to synthesize endothelin (ET)-1. AM have been known to express prepro ET-1 mRNA. Thus, we hypothesized that thrombin stimulates AM so as to synthesize ET. Surgically resected human lungs were irrigated by saline to remove intravascular blood, then saline was instilled into the bronchus, and the fluids were recovered. AM were separated by Percoll density centrifugation (density, 1.060 g/ml). AM were resuspended in culture medium without FCS in the presence or absence of thrombin. ET was synthesized by thrombin in a concentration-dependent manner, and the amounts of ET synthesized by thrombin (10 U/ml) were equivalent to those by LPS (10 microg/ml). Dexamethasone (10(-6)-10(-10) M), IL-4 (100 U/ml), and TGF-beta (10 ng/ml) significantly suppressed the ET synthesis by thrombin (p < 0.05). In contrast, 1,25-dihydroxyvitamin D3 (10(-8)-10(-10) M) enhanced the ET synthesis up to approximately 300%. The analysis using high pressure liquid chromatography revealed that AM-derived ET consists of ET-1, ET-2, and ET-3. Major constituents were ET-2 and ET-1, and the ratio of ET-2/ET-1 was 1.7 +/- 0.4 (mean +/- SE). These results indicate that thrombin is a potent agonist for AM to synthesize ET.

Cells, Cultured↗
PubMed1997

The glycoproteins that occur in the colloids of senescent porcine pituitary glands are clusterin and glycosylated albumin fragments.

Periodic acid-Schiff-positive colloids occur in the pituitary glands of many vertebrates. These colloids are surrounded by folliculo-stellate (FS) cells and increase dramatically in number and size with age. The characterization of these colloidal substances is necessary for the establishment of FS cell function. In a previous study we showed that these colloids in the senescent pituitary contain a number of glycoproteins; however, the characterization of these glycoproteins has not yet been carried out. To characterize the pituitary colloid, we purified the colloids by centrifugation and Percoll gradient. The isolated colloids were unable to be solubilized in a regular buffer since they were very dense. However, solubilization was accomplished in sodium dodecyl sulfate (SDS) lysis buffer following treatment with exoglycosidase. Analysis using SDS-polyacrylamide gel electrophoresis showed that the porcine pituitary colloids contain four proteins, CP26, CP40, CP48 and CP60, whose estimated molecular weights were 26 kD, 36-43 kD, 48 kD, and 60 kD, respectively. Staining with concanavalin A (ConA) showed that both CP26 and CP40 were glycoproteins. Moreover, N-terminal amino acid sequencing and Western blot analysis revealed CP26, CP48 and CP60 to be porcine albumin while CP40, the major protein in the colloids, proved to be clusterin. Therefore we report here that the glycoproteins that occur in senescent porcine pituitary colloids are of two types, namely, albumin fragments and clusterin.

Aging↗
PubMed1997

Overexpression of N-acetylglucosaminyltransferase III disrupts the tyrosine phosphorylation of Trk with resultant signaling dysfunction in PC12 cells treated with nerve growth factor.

beta-1,4-N-Acetylglucosaminyltransferase III (GnT-III: EC 2.4.1.144) is a pivotal glycosyltransferase which participates in branch formation by catalysis of the synthesis of a bisecting GlcNAc structure in N-glycans. These structures are thought to be one of the unique features of the N-glycans of neural tissues. To examine the intracellullar role of GnT-III expression and its product in neural cells, its gene was overexpressed in rat pheochromocytoma PC12 cells which normally express a low level of GnT-III. In the GnT-III gene-transfected cells, lectin blot analysis showed that some glycoproteins showed increased levels of bisecting GlcNAc structures. Following treatment with nerve growth factor (NGF) the control cells showed neurite outgrowth for differentiation whereas the transfectants showed no morphological response or change in the rate of cell growth. Transient tyrosine phosphorylation of the Trk/NGF receptor was detected at 5-15 min after NGF treatment in control cells, but not detected in the GnT-III gene-transfected cells despite the intact binding of NGF to the cells. Moreover the dimerization of Trk with NGF treatment was not induced in the GnT-III transfectant as compared with the dimerization seen in control cells. These results indicate that overexpression of GnT-III gene in PC12 cells affects some functions of glycoprotein receptors such as Trk by alteration of N-glycan structures, and results in changes in the intracellular signaling pathway of tyrosine phosphorylation modified by NGF.

Animals↗
PubMed1997

Role of dynamic MRI in the evaluation of head and neck cancers treated with radiation therapy.

PURPOSE: To study the usefulness of dynamic magnetic resonance imaging in the evaluation of head and neck cancers treated with radiation therapy. METHODS AND MATERIALS: Seventy-six patients (58 males and 18 females; ages 20-82) with head and neck cancers (10 nasopharyngeal carcinomas, 22 mesopharyngeal carcinomas, 10 hypopharyngeal carcinomas, 16 oral cavity carcinomas, 11 lingual carcinomas, and 7 laryngeal carcinomas) were treated by radiation therapy combined with concomitant low-dose cisplatinum. Magnetic resonance imaging (MRI) was performed before and 2 weeks after the irradiation in all cases. After bolus administration of gadopentetate dimeglumine (Gd-DTPA) (0.1 mmol kg), images were obtained every 30 s (repetition time 200 ms, echo time 16 ms) using a 1.5 or 0.5-T superconductive unit. Biopsy or surgery was performed after radiation therapy and the histologic findings were correlated with the MRI findings (T1, T2, dynamic, and enhanced T1). RESULTS: Complete remission, partial response, and no response were obtained in 18, 36, and 7 cases, respectively. Dynamic MRI correctly diagnosed 17 of the 18 complete remission cases, 33 of the 36 partial response cases, and all of the 7 no-response cases. The accuracy of dynamic MRI, T1-weighted image, T2-weighted image, and Gd-enhanced T1-weighted image was 94.4%, 68%, 82%, and 86%, respectively. CONCLUSION: Dynamic MRI proved to be useful in the evaluation of the radiation therapy of head and neck cancers.

Adult↗
PubMed1997

Peroxynitrite induces the conversion of xanthine dehydrogenase to oxidase in rabbit liver.

Effect of peroxynitrite (ONOO-) on the conversion of xanthine dehydrogenase to oxidase in rabbit liver was examined. ONOO- (25-200 microM) induced the conversion of xanthine dehydrogenase to oxidase in a dose-dependent manner. The addition of hydroxyl radical scavengers (mannitol and dimethyl sulfoxide) gave no alteration in the ONOO(-)-induced conversion of xanthine dehydrogenase to oxidase, implying that the action of ONOO- is not due to hydroxyl radicals which may be formed from ONOO-. The experiment utilizing dithiothreitol also revealed that the action of ONOO- might be due to oxidation of sulfhydryl group of xanthine dehydrogenase. These results suggest that ONOO- has the potential to convert xanthine dehydrogenase to oxidase, and that this effect may be correlated with cytotoxic actions of ONOO-.

Animals↗
PubMed1997

Increased glucagon action on lactate gluconeogenesis in perfused liver of dexamethasone-treated rats.

To clearly understand the hyperglycemic action of glucocorticoids, we studied the action of glucagon on lactate gluconeogenesis in the liver of rats 7 days after adrenalectomy and after treatment with 1 mg/kg dexamethasone for 7 days. The liver was isolated and cyclically perfused at 20 ml/min with 25 ml of perfusion medium containing 5 mM lactate, [U-14C]lactate, and 0-100 ng/ml glucagon. In the absence of glucagon, incorporation of [14C]lactate into glucose carbon 1 did not change significantly in the adrenalectomized rat liver (1.66 +/- 0.12% of total radioactivity for 5 min) and increased in the dexamethasone-treated rat liver (3. 61 +/- 0.54%, P < 0.01) compared to the normal rat liver (1.99 +/- 0. 28%). The response of lactate gluconeogenesis to glucagon was extremely blunted in the adrenalectomized rat liver and was much larger in the dexamethasone-treated rat than in the normal rat liver (at a glucagon concentration of 100 ng/ml, 2.13 +/- 0.33, 8.55 +/- 1. 06, and 4.61 +/- 0.53% for 5 min, respectively). Glucagon binding to liver plasma membrane was not changed by adrenalectomy and was decreased by dexamethasone treatment. These results suggest that glucocorticoids induce hyperglycemia by increasing the response to glucagon, together with the high basal activity of hepatic gluconeogenesis. In addition, these effects do not occur through changes in glucagon binding to receptors.

Animals↗
PubMed1997

Ballistocardiogram of avian eggs determined by an electromagnetic induction coil.

As an avian embryo grows within an eggshell, the whole egg is moved by embryonic activity and also by the embryonic heartbeat. A technical interest in detecting minute biological movements has prompted the development of techniques and systems to measure the cardiogenic ballistic movement of the egg or ballistocardiogram (BCG). In this context, there is interest in using an electromagnetic induction coil (solenoid) as another simple sensor to measure the BCG and examining its possibility for BCG measurement. A small permanent magnet is attached tightly to the surface of an incubated egg, and then the egg with the magnet is placed in a solenoid. Preliminary model analysis is made to design a setup of the egg, magnet and solenoid coupling system. Then, simultaneous measurement with a laser displacement measuring system, developed previously, is made for chicken eggs, indicating that the solenoid detects the minute cardiogenic ballistic movements and that the BCG determined is a measure of the velocity of egg movements.

Animals↗
PubMed1997

Smoking cessation increases the resistance of low-density lipoprotein to oxidation.

The effects of smoking cessation on the susceptibility to oxidation of low-density lipoprotein (LDL) was investigated in 14 men who quit smoking for 3 months. LDL was isolated and susceptibility of LDL to V-70 (4-methoxy-2,4-dimethylvalerinitrile)-mediated oxidation was assessed by measuring conjugated diene production at 234 nm, the lag phase being a measure of the resistance of LDL to oxidation. The mean duration of the lag phase became 1.9-fold longer after 3 months (P < 0.001). The result suggests that the increase in resistance of LDL to oxidation contributes to the reduction of the risk of coronary heart disease by smoking cessation.

Adult↗
PubMed1997

The pituitary folliculo-stellate cell line TtT/GF augments basal and TRH-induced prolactin secretion by GH3 cell.

Numerous functions have been ascribed to anterior pituitary folliculo-stellate (FS) cells. Recently, we established a pituitary FS-like cell line (TtT/GF) from an isologously transplantable pituitary thyrotropic tumor. Here we investigated the effect of FS cells on prolactin (PRL) secretion by using a co-culture system between TtT/GF cells and GH3 cells. We observed that co-culture with TtT/GF cells significantly increased basal PRL secretion without affecting the growth rate of GH3 cells. Further, addition of anti-cytokine-induced neutrophil chemoattractant (CINC) antibody partially reduced PRL secretion in co-culture of GH3 cells with 10% TtT/GF cells. Co-culture with 5% TtT/GF cells also markedly enhanced the PRL response to thyrotropin-releasing hormone (TRH) by GH3 cells, while addition of anti-CINC antibody had no effects on the enhanced PRL response to TRH. These results suggest that FS cells augment basal PRL secretion and enhance the secretory response of PRL to TRH by a paracrine mechanism.

Animals↗
PubMed1997

Pituitary folliculo-stellate-like cell line produces a cytokine-induced neutrophil chemoattractant.

In previous studies, we reported that cytokine-induced neutrophil chemoattractant (CINC) was produced in the pituitary gland. Here we investigated the possibility of detection of CINC immunoreactivity in the pituitary folliculo-stellate (FS)-like cell line (TtT/GF). Intense immunoreactivity was observed by immunocytochemistry in the cytoplasm and cell processes of TtT/GF cells. CINC immunoreactivity was detected by an enzyme-linked immunosorbent assay in as little as 3 h after conditioning the medium with TtT/GF cells, and it increased significantly in a time-dependent manner during the first 24 h of the culture. This immunoreactivity could be induced by lipopolysaccharide and tumor necrosis factor-alpha in a time- and dose-dependent manner. We also observed the expression of CINC mRNA in TtT/GF cells and LPS increased CINC mRNA accumulation in TtT/GF cells. These findings indicate that CINC produced by FS cells may play a role as a paracrine factor of the anterior pituitary gland, and TtT/GF will provide a useful model system for studying the regulation of CINC secretion by FS cells.

Animals↗
PubMed1997

Measurement of reaction rate distributions in a plastic phantom irradiated by 40- and 65-MEV quasi-monoenergetic neutrons.

Reaction rate distributions in a plastic phantom were measured with solid state nuclear track detectors and a fission counter for 40- and 65-MeV quasi-monoenergetic neutrons generated by the 7Li(p,n) reactions with 43- and 68-MeV protons at AVF cyclotron of Japan Atomic Energy Research Institute. Measured distributions were compared with calculated ones.

Cyclotrons↗
PubMed1997

Increased natural killer resistance to cyclosporine A by continuous doses of dexamethasone in rats.

There is a controversy on the effects of physiological levels of glucocorticoids on natural killer (NK) cytotoxity. Therefore, the effects of exogenously administered dexamethasone on NK cytotoxity in 8-week-old male, Fischer 344 rats were studied. We suppose that the reason for the controversy is insufficient sensitivity of the ordinal radioactive chromium-release assay for normal healthy subjects or animals. Therefore, we developed a new index, a resistance to artificial immunosuppressor, cyclosporine A (CsA) using rat NK activity as an indicator, and named this index, increased resistance to immunosuppressor (IRIS). After some basic, characterizing studies, authors confirmed the fact that continuous doses of dexamethasone (DEX) attenuated NK suppression of CsA. In protocol 4, 18 rats were randomly divided into three groups: the first (DEX + CsA) was injected for 5 days with 0.1 mg DEX/kg/day and a single dose of CsA on the final day, intraperitoneally; the second (SAL + CsA) was treated with an equal volume of saline and CsA; the third (DEX + SAL) was treated with DEX but not CsA. The IRIS in NK activity was increased significantly (P < 0.01) with 5 days injection of DEX. These results demonstrated that physiological, and continuous dosage of glucocorticoids stimulated IRIS in NK activity in rats, and this suggests that appropriate stimuli through the hypothalamic-adrenal axis might be acting, at least, as a defence against immune collapses or dysfunctions.

Animals↗