Biomedical subjects

N Toyoda

Publications and source records attributed to N Toyoda.

At least 19 recordsLinked to original sources

PubMed2003

Expression and cytoprotective effect of protease-activated receptor-1 in gastric epithelial cells.

Thrombin is a serine protease involved in many physiological functions and its receptor. the protease-activated receptor-1 (PAR-1), has a wide tissue distribution. We hypothesized that PAR-1 is expressed in gastric epithelial cells and that thrombin can modulate defence mechanisms through PAR-1. The rat gastric epithelial cell line (RMG1) and gastric biopsy specimens from gastritis patients were used in the study. Reverse transcriptase polymerase chain reaction analysis showed that the thrombin receptors PAR-1, PAR3 and PAR-4 are expressed by RGM1 gastric epithelial cell line. Immunohistochemical and electron microspcopic studies also showed PAR-1 expression in human gastric epithelial cells. Thrombin stimulated the secretion of mucin and prostaglandin E2 (PGE2) formation in RGM1 cells in a dose-dependent manner. PAR-1 agonist also stimulated PGE2 formation. In addition, thrombin significantly increases the expression of the PGE2 receptors EP2-R and EP4-R in RGM1 cells. In conclusion, the results of the present study showed for the first time that gastric epithelial cells express thrombin receptors and that these receptors may play a protective role in the gastric mucosa.

Animals↗

PubMed2003

Salvage chemotherapy with a combination of irinotecan hydrochloride and mitomycin C in platinum- and paclitaxel-resistant epithelial ovarian cancer: case reports.

INTRODUCTION: The efficacy and toxicity of salvage chemotherapy with a combination of irinotecan hydrochloride (CPT-11) and mitomycin C (MMC) for platinum-and paclitaxel-resistant epithelial ovarian cancer are reported. CASE REPORT: Three consecutive patients with platinum- and paclitaxel- resistant epithelial ovarian cancer were treated with 120 mg/m2 of CPT-11 (days 1 and 15) and 7 mg/m2 of MMC (days 1 and 15) every four weeks. In all three cases partial responses were achieved and overall survivals were 17 months or longer. Most of the adverse side-effects were manageable. CONCLUSIONS: This regimen could be administered even in heavily pretreated patients with platinum- and paclitaxel- resistance. Phase I and II studies are needed to confirm the feasibility of this treatment. The efficacy of most salvage treatments for platinum- and paclitaxel-resistant epithelial ovarian cancer is disappointing, and our cases might be of interest from the perspective of treating platinum- and paclitaxel-resistant ovarian cancer.

Aged↗

PubMed2002

Gene transfer of Fc-fusion cytokine by in vivo electroporation: application to gene therapy for viral myocarditis.

Among a number of techniques for gene transfer in vivo, the direct injection of plasmid DNA into muscle is simple, inexpensive and safe. Although combining direct DNA injection with in vivo electroporation increases the efficiency of gene transfer into muscle, applications of this method have remained limited because of the relatively low expression level. To overcome this problem, we developed a plasmid vector that expresses a secretory protein as a fusion protein with the noncytolytic immunoglobulin Fc portion and used it for electroporation-mediated viral interleukin 10 (vIL-10) expression in vivo. The fusion cytokine vIL-10/mutFc was successfully expressed and the peak serum concentration of vIL-10 was almost 100-fold (195 ng/ml) higher than with a non-fusion vIL-10 expression plasmid. The expressed fusion cytokine suppressed the phytohemagglutinin-induced IFN-gamma production by human peripheral blood mononuclear cells and decreased the mortality in a mouse viral myocarditis model as effectively as vIL-10 expression. These results demonstrate that the transfer of plasmid DNA expressing a noncytolytic Fc-fusion cytokine is useful to deliver enhanced levels of cytokine without altering general biological activities. This simple and efficient system should provide a new approach to gene therapy for human diseases and prove very useful for investigating the function of newly discovered secretory protein genes.

Animals↗

PubMed2002

Analysis of glucose tolerance in twin gestations using an oral glucose load.

The effect of twin gestation on carbohydrate metabolism was evaluated using a 75 g oral glucose tolerance test (75 g OGTT). A 75 g OGTT was performed in 63 twin gestations and 3 791 singleton gestations during the third trimester. Plasma glucose concentrations were measured in the pregnant women under fasting conditions as well as 30 min, 1 h, and 2 h after ingestion of glucose (75 g oral load), and serum insulin concentrations were measured in fasting and 30 min post-ingestion samples. Women with twin gestations showed significantly lower plasma glucose concentrations during fasting and 30 min after the glucose load in the samples taken than those with singleton gestations. No significant difference in serum glucose concentrations was found in the other specimens. There were no cases of gestational diabetes mellitus in our study. Although women with twin gestations demonstrated lower plasma glucose concentrations than women with singleton gestations, the difference observed was subtle. We could not find any significant differences in these plasma glucose values as used to define a pathologic OGTT between twin and singleton pregnancies, with the exception of the fasting value.

Adult↗

PubMed2001

Serial analysis of gene expression in chronic hepatitis C and hepatocellular carcinoma.

Hepatitis C virus (HCV) causes chronic hepatitis C (CH-C) and is epidemiologically linked with the occurrence of hepatocellular carcinoma (HCC). To elucidate the comprehensive gene expression profiles of CH-C and HCC, serial analysis of gene expression (SAGE) libraries were made from CH-C and HCC tissues of a patient, and compared with a reported SAGE library of a normal liver (NL). Scatter plots of the distribution of tags from the HCC library exhibited the existence of many differentially expressed genes compared with those from the CH-C and NL libraries. Up-regulation of IFN-gamma inducible genes and oxidative stress-inducible genes were identified in both the CH-C and HCC libraries, and some unpublished new genes were specifically up- or down-regulated in the HCC library. This genome-wide scanning study discloses the molecular portraits of CH-C and HCC, and provides novel candidate genes that should help clarify the mechanism of hepatocarcinogenesis in the chronically HCV-infected liver.

Base Sequence↗

PubMed2001

Prognostic value of the colposcopic tumor size in stage IB squamous cervical cancer.

BACKGROUND AND OBJECTIVES: To determine the prognostic significance of the colposcopic tumor size in the management of cervical cancer. METHODS: Clinicopathological analysis was performed in 751 consecutive patients with stage IB squamous cervical cancer who were surgically treated in a single institute. The colposcopic tumor size was measured postoperatively on surgical specimens. Univariate and multivariate analyses were performed to determine the prognostic significance of various pathological factors. RESULTS: Among the pathological factors examined, lymph node metastasis, parametrial extension, deep stromal invasion, vessel permeation, endometrial extension, and colposcopic tumor size were found to be prognostic factors in univariate analysis, whereas multivariate analysis has confirmed that only three factors, i.e., lymph node metastasis, parametrial involvement, and colposcopic tumor size were independently associated with the disease-free interval. CONCLUSIONS: These results indicate that the colposcopic tumor size is an independent prognostic factor in squamous cervical cancer and can be used as an indicator of treatment options.

Adult↗

PubMed2001

A prospective randomized comparison of routine buserelin acetate and a decreasing dosage of nafarelin acetate with a low-dose gonadotropin-releasing hormone agonist protocol for in vitro fertilization and intracytoplasmic sperm injection.

OBJECTIVE: To compare the efficacy of a draw-back nafarelin acetate protocol with routine buserelin acetate administration for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). DESIGN: Prospective clinical study. SETTING: Mie University School of Medicine, Tsu, Mie, Japan. PATIENT(S): One hundred sixty-nine women treated with IVF and 183 women treated with ICSI. INTERVENTION(S): Nafarelin acetate and buserelin acetate in ovarian hyperstimulation in IVF and ICSI were administered. MAIN OUTCOME MEASURE(S): The concentrations of estradiol (E(2)), FSH, LH, gonadotropin dosages; the number of oocytes retrieved, oocytes fertilized, and embryos; and pregnancy rates. RESULT(S): A prospective study was conducted with 44 cycles for 34 couples with nafarelin acetate (group 1) and 47 cycles for 40 couples with buserelin acetate (group 2) with a long IVF protocol; 68 cycles for 46 couples with nafarelin acetate (group 3) and 56 cycles for 39 couples with buserelin acetate (group 4) with a short IVF protocol; 39 cycles for 32 couples with nafarelin acetate (group 5) and 50 cycles for 30 couples with buserelin acetate (group 6) with a long ICSI protocol; and 87 cycles for 60 couples with nafarelin acetate (group 7) and 81 cycles for 61 couples with buserelin acetate (group 8) with a short ICSI protocol. Patients were randomized to receive either full-dose nafarelin acetate (200 microg b.i.d.) treatment for 7 days followed by half-dose nafarelin acetate (200 microg daily) or buserelin acetate (300 microg t.i.d.). There were no statistically significant differences in baseline concentrations of E(2) and FSH, concentrations of E(2), P4, FSH, LH on hCG administration, gonadotropin dosage, the number of oocytes retrieved and embryos transferred, or pregnancy rates between groups 1 and 2, groups 3 and 4, groups 5 and 6, and groups 7 and 8. CONCLUSION(S): Full-dose nafarelin acetate treatment for 7 days followed by half-dose nafarelin acetate ("draw-back" protocol) is an effective new protocol for IVF and ICSI.

Abortion, Spontaneous↗

PubMed2001

Type 2 iodothyronine deiodinase expression is upregulated by the protein kinase A-dependent pathway and is downregulated by the protein kinase C-dependent pathway in cultured human thyroid cells.

Type 1 and 2 iodothyronine deiodinases (D1 and D2) catalyze thyroxine (T4) activation. In human thyroid, unlike rodents', both D1 and D2 are expressed. We have investigated the effects of thyrotropin (TSH), dibutyryl cyclic adenosine monophosphate [(Bu)2cAMP] (an activator of protein kinase A [PKA]), 12-O-tetradecanoylphorbor 13-actate (TPA) (an activator of protein kinase C [PKC]), T4, and triiodothyronine (T3) on the D2 mRNA levels and activity in cultured human thyroid cells. D2 mRNA levels were increased by TSH and (Bu)2cAMP, and the increment was faster and greater than that of D1 mRNA levels. The increment of the maximum velocity (Vmax) value for D2 by (Bu)2cAMP stimulation was similar to that of D2 mRNA levels, suggesting that (Bu)2cAMP enhances D2 activity mainly at the pretranslational level. Cycloheximide, a protein synthesis inhibitor, partially inhibited the increase of D2 mRNA levels by (BU)2cAMP, suggesting that de novo protein synthesis-dependent pathways are involved. TPA suppressed the D2 mRNA levels in the presence of (Bu)2cAMP. However, T3 and T4 did not significantly change the D2 mRNA levels and activity. In conclusion, D2 expression in human thyroid cells is more rapidly and strongly upregulated by the PKA pathway than D1 expression, and is downregulated by the PKC pathway.

Bucladesine↗

PubMed2001

Comprehensive gene expression profile of human activated T(h)1- and T(h)2-polarized cells.

In response to antigen stimulation, T(h) cells differentiate into two types of effector cells, T(h)1 and T(h)2. T(h)1 cells predominantly mediate cellular immunity, whereas T(h)2 cells induce humoral allergic responses. We have conducted here serial analysis of gene expression (SAGE) in human activated T(h)1- and T(h)2-polarized cells from cord blood. SAGE analysis of 64,510 tags (32,219 and 32,291 tags from T(h)1 and T(h)2 cells respectively) allowed identification of 22,096 different transcripts. In activated T(h)1 cells, many of the known genes (12 genes, P: < 0.01; 56 genes, P: < 0.05), including genes encoding IFN-gamma, lymphotactin, osteopontin, MIP-1alpha, MIP-1beta, perforin, beta-catenin and CD55, are highly expressed. On the other hand, in activated T(h)2 cells rather limited numbers of known genes (four genes, P: < 0.01; 10 genes; P: < 0.05), such as genes encoding FUS, ILF-2, IL-13 and E2-EPF, are found to be selectively expressed. The comprehensive identification of genes selectively expressed in human activated T(h)1 or T(h)2 cells should contribute to our understanding of the molecular basis of T(h)1/T(h)2-dominated human diseases and may provide genetic information to diagnose these diseases.

Cells, Cultured↗

PubMed2001

Comparison of piezo-assisted micromanipulation with conventional micromanipulation for intracytoplasmic sperm injection into human oocytes.

OBJECTIVE: To compare the efficacy of piezo-assisted micromanipulation with conventional micromanipulation for intracytoplasmic sperm injection (ICSI) into oocytes in patients with impaired semen parameters and no success with in vitro fertilization (IVF). STUDY DESIGN: A retrospective randomized study was conducted on 204 cycles for 104 couples with piezo-assisted ICSI and 122 cycles for 96 couples with conventional ICSI. Piezo-assisted ICSI consists of two steps, namely penetration of the zona pellucida alone with a piezo-pulse and then puncturing of the oolemma with a light negative pressure without piezo, as with conventional ICSI. The tips of injection pipettes were prepared after pulling by breakage with a scalpel under a microscope, so that the inner diameter at and near the tip was 5 microm, as for conventional ICSI. RESULTS: Piezo-assisted ICSI demonstrated significantly more favorable results, with a fertilization rate of 90.3% (conventional ICSI: 83.1%, p < 0.01) and a cleavage rate of 88.1% (conventional ICSI: 84.6%, p < 0.01). CONCLUSION: Piezo-assisted ICSI is easy to incorporate a spermatozoa exactly into the ooplasm with little deformation of the oocyte during insertion. Piezo-assisted ICSI can be used effectively for human oocytes to improve the fertilization, cleavage rates.

Adult↗

PubMed2000

Comprehensive gene expression profile of LPS-stimulated human monocytes by SAGE.

Monocytes play a pivotal role in various human infectious and inflammatory diseases. To reveal a whole picture of pathophysiologic function of activated human monocytes, this study used the serial analysis of gene expression (SAGE) procedure in lipopolysaccharide (LPS)-stimulated human monocytes. A total of 35 874 tags corresponding to more than 12 000 different transcripts were sequenced. Comparison of gene expression profile with that of resting monocytes revealed the LPS-inducible gene expression profile. Many cytokines and chemokines, including interleukin (IL)-6, IL-1alpha, IL-1beta, tumor necrosis factor (TNF)-alpha, macrophage inflammatory protein (MIP)-1beta, MIP-2beta, MIP-2alpha, liver and activation-regulated chemokine (LARC), MIP-1alpha, thymus and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC), regulated on activation, normal T cell expressed and secreted (RANTES), growth-regulated oncogene (GRO) alpha, and IL-8, were observed in the highest inducible transcripts. Other genes encoding plasminogen activator inhibitor type 2 (PAI-2), Hc-gp39, apolipoproteins, malate dehydrogenase, matrix metalloproteinase-9 (MMP-9), and cyclooxygenase (COX2) were also highly elevated in LPS-stimulated monocytes. Moreover, up-regulation of Naf1beta, IL-7 receptor, adenosine receptor A2a, and many novel genes was newly identified. These results suggest that the LPS-inducible gene products may be involved in cell activation and migration, angiogenesis, tissue remodeling, and metabolism, and thus may orchestrate the inflammatory reactions. On the other hand, the expression of numerous sets of novel genes was discovered to be down-regulated on LPS stimulation. This study represents the first comprehensive analysis of LPS-inducible gene expression in human monocytes and provides tremendous novel information for the function of LPS-activated monocytes and targets for diagnosing, monitoring, and treating sepsis and various human infectious and inflammatory diseases.

Base Sequence↗

PubMed2000

Identification of genes specifically expressed in human activated and mature dendritic cells through serial analysis of gene expression.

Dendritic cells (DCs) are professional antigen-presenting cells in the immune system and can be generated in vitro from hematopoietic progenitor cells, DC precursors, and monocytes in peripheral blood. Serial analysis of gene expression (SAGE) was conducted in lipopolysaccharide (LPS)-stimulated mature and activated DCs (MADCs) derived from human blood monocytes. A total of 31 837 tag sequences from an MADC cDNA library represented 10 962 different genes, and these data were compared with SAGE data for monocyte-derived immature DCs (IMDCs). Many of the genes, such as germinal center kinase-related protein kinase, cystatin F, interferon (IFN)-alpha-inducible protein p27, EBI3, HEM45, actin-bundling protein, ELC, DC-LAMP, serine/threonine kinase 4, and several genes in expressed sequence tags, were differentially expressed in MADCs, and those encode proteins related to cell structure, antigen-processing enzymes, chemokines, and IFN-inducible proteins. The profile of MADCs was also compared with that of LPS-stimulated monocytes. The Epstein-Barr virus-induced gene 3 and IFN-alpha-inducible protein p27 are newly identified to be specifically and highly expressed in MADCs, but not in LPS-stimulated monocytes. The comprehensive identification of specific genes expressed in human IMDCs and MADCs should provide candidate genes to define heterogeneous subsets as well as the function and maturation stage of DCs.

Antigen Presentation↗

PubMed2000

WISP-2 as a novel estrogen-responsive gene in human breast cancer cells.

In order to search for novel estrogen-responsive genes, we performed serial analysis of gene expression (SAGE) for estrogen-treated MCF-7 human breast cancer cells. SAGE analysis of 31,000 and 30,856 tags from non-treated and 17 beta-estradiol (E2)-treated cells for 24 h, respectively, facilitated the identification of 15,037 different transcripts. Comparison of these two SAGE libraries indicated a remarkable similarity in expression profiles. Among the identified transcripts, four genes were found to be markedly increased for E2-treated cells compared with control cells. Three of the transcripts were cathepsin D, pS2 and high mobility group 1 protein, which have been described as estrogen-inducible genes. The fourth gene was WISP-2 (Wnt-1 inducible signaling pathway protein 2) which has recently been reported as an up-regulated gene in the mammary epithelial cell line C57 MG transformed by the Wnt-1 oncogene. The increase in WISP-2 mRNA was completely prevented by co-incubation with a pure anti-estrogen ICI 182,780, but not by coincubation with cycloheximide, indicating that WISP-2 is directly regulated by the estrogen receptor. The WISP-2 gene was also induced by treating with environmental estrogens, such as bisphenol-A or nonylphenol. This study represents the first comprehensive gene expression analysis of estrogen-treated human breast cancer cells.

Air Pollutants, Occupational↗

PubMed2000

Comprehensive gene expression profile of a normal human liver.

To investigate the gene expression profile of a normal human liver, we performed serial analysis of gene expression (SAGE), which allows the quantitative and simultaneous analysis of thousands of genes expressed in tissue. Polyadenylated RNA was obtained from a bulk normal human liver sample and SAGE was performed. Reverse transcriptase-polymerase chain reaction (RT-PCR) was also performed in each of 3 different normal liver samples to evaluate the validity of the profile in each individual. A total of 30,982 tags were sequenced, 8,596 of which were unique. The genes highly expressed in the normal liver were those encoding plasma proteins (>21.8% of total transcripts), cytoplasmic proteins (>8.6%), enzymes (>4.8%), protease inhibitors (>1.7%), complements (>1.1%), and coagulation factors (>0.75%). About 13.9% of all transcripts encoded genes not reported in GenBank thus far. This study identifies candidate genes to be examined in relation to various human liver diseases, including viral hepatitis, liver cirrhosis, and hepatocellular carcinoma.

Adult↗

PubMed2000

A novel human HER2-derived peptide homologous to the mouse K(d)-restricted tumor rejection antigen can induce HLA-A24-restricted cytotoxic T lymphocytes in ovarian cancer patients and healthy individuals.

A mouse HER2-derived peptide, HER2p63 (A) (TYLPANASL), can induce K(d)-restricted mouse cytotoxic T lymphocytes (CTL) and also function as a tumor rejection antigen in an in vivo assay. Since the anchor motif of mouse K(d) for peptide binding has much similarity to that of human HLA-A2402, we asked if human HER2p63 (T) (TYLPTNASL) could induce HER2-specific CTL in HLA-A2402-positive individuals. Peripheral blood mononuclear cells (PBMC) of HLA-A2402-positive individuals were sensitized in vitro with HER2p63-pulsed autologous dendritic cells prepared from PBMC. CTL clone derived from these specifically lysed HER2-expressing cell lines bearing HLA-A2402. Cytotoxic activity of the CTL clone against the HER2-expressing cell line bearing HLA-A2402 was blocked by antibodies against CD3, CD8, HLA-A24 or MHC class I, and was also inhibited by the addition of excess HER2p63-pulsed C1R bearing HLA-A2402. Killer cells were generated from PBMC of seven healthy individuals and five ovarian cancer patients, all of HLA-A2402 type, by in vitro sensitization with HER2p63-pulsed autologous antigen presenting cells. These killer cells selectively lysed HER2-expressing SKOV3 transfected with HLA-A2402 cDNA, indicating high immunogenicity of HER2p63 in all 12 individuals examined.

Animals↗

PubMed2000

Real-time ultrasonographically guided transvaginal instillation for intraperitoneal chemotherapy of ovarian carcinoma.

OBJECTIVE: To describe the techniques we have developed to instill chemotherapeutic agents into the pelvic cavity using real-time transvaginal ultrasonographic guidance. METHODS: This was used for intraperitoneally administered chemotherapy in 11 patients with ovarian carcinoma. RESULTS: A total of 38 instillations were done in these patients. The average time for instillation was 23.9 min. No adhesions were associated with this chemotherapy. As for complications, hematuria occurred during 2 (5.3%) instillations. No other complications were observed. CONCLUSION: Real-time ultrasonographically guided transvaginal intraperitoneal chemotherapy can be performed without any catheter systems. This is a safe, reliable, convenient, repeatable, and inexpensive procedure.

Adult↗

PubMed2000

Expression of glucose transporter 4 mRNA in adipose tissue and skeletal muscle of ovariectomized rats treated with sex steroid hormones.

The effects of 17beta-estradiol (E) and/or progesterone (P) on glucose transporter 4 (GLUT4) expression in the adipose tissue and skeletal muscle of ovariectomized female rats were studied. The Sprague-Dawley rats received daily subcutaneous injections of various doses of E and/or P for 7 days (n=5-6 per dose). The expression of GLUT4 mRNA was assessed by performing ribonuclease protection assays. GLUT4 protein levels were assessed by Western blotting assays. The adipose tissue levels of GLUT4 mRNA were reduced by the administration of 50 microg E, which resulted in unphysiologically high serum E concentrations. Although the GLUT4 mRNA levels did not change after the administration of 10 microg E or 5 mg P, they were reduced significantly to approximately half the control group level by the administration of both hormones (p <0.01). The skeletal muscle GLUT4 mRNA levels were not changed significantly by hormone treatment. These findings suggest that E and P may be involved in the regulation of GLUT4 mRNA expression in adipose tissue.

Adipose Tissue↗

PubMed2000

Relation of rat fetal body weight to the relative expression of insulin genes I and II.

This study investigates the relation between the ratio of the abundance of mRNAs encoded by insulin genes I and II (insulin I/II mRNA ratio) and the weight of rat fetuses on gestational day 20. Total RNA was extracted from the pancreas of the fetuses with the maximum and minimum body weight in each litter on gestational day 20. The amount of insulin mRNAs I and II in each RNA preparation was determined by reverse transcription-polymerase chain reaction (RT-PCR) analysis and restriction enzyme digestion. The maximum and minimum weight of the fetuses were 4.07 +/- 0.23 and 3.23 +/- 0.34 g, respectively (N = 18, P < .01) and the corresponding insulin I/II mRNA ratios were 3.65 +/- 0.55 and 1.42 +/- 0.21 (N = 18, P < .01). Furthermore, the insulin I/II mRNA ratio correlated significantly with fetal weight (r = .526, P < .05). These results suggest that the relative expression of insulin genes I and II may affect the extent of fetal growth.

Animals↗