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1.
Transplant Proc ; 38(10): 3211-3, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17175226

ABSTRACT

Nitric oxide (NO) is a multifunctional molecule in a variety of physiologic and pathologic processes. Its precise effect on human T lymphocyte responses against alloantigens are not yet fully known, although it has been reported that NO is antiproliferative and can cause apoptosis in several cell types. To address these issues, we analyzed the effects of an NO donor on mixed lymphocyte cultures (MLC) and on apoptosis induction in T lymphocytes activated with alloantigens. NOC 18 was used as an NO donor. The MLC was performed with human peripheral blood mononuclear cells isolated from healthy volunteers. Cell division and interleukin (IL)-2 production were measured with CFSE labeling and an EIA kit, respectively. After cells were incubated with NOC 18 for 24 hours, DNA fragmentation was assessed using the diphenylamine assay. Pre-culture of cells with NOC 18 for 24 hours resulted in significant inhibition of cell proliferation and IL-2 production in MLC. NOC 18 induced DNA fragmentation of cells harvested from an MLC following 7 days of the culture, in a dose-dependent manner, whereas it never exerted any influence on DNA fragmentation of freshly isolated cells. A chemical NO donor, NOC-18, may have immunosuppressive ability when treatment of responder cells occurs before the beginning of the MLC and may induce apoptosis of alloantigen-activated T lymphocytes.


Subject(s)
Lymphocyte Activation/immunology , Lymphocytes/immunology , Nitric Oxide/biosynthesis , T-Lymphocytes/immunology , Apoptosis , Humans , Interleukin-2/biosynthesis , Lymphocyte Culture Test, Mixed
2.
Clin Nephrol ; 62(3): 180-4, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15481849

ABSTRACT

Gitelman syndrome (GS, MIM 263800) is an inherited disorder characterized by metabolic alkalosis with hypokalemia, hypomagnesemia, and hypocalciuria. The genetic abnormalities causing GS are known to lie in the thiazide-sensitive NaCl cotransporter (TSC), which is expressed in the distal tubule of the kidney. The TSC gene, located at chromosome 16, consists of 26 exons and encodes the protein containing 12 putative transmembrane domains with long intracellular amino and carboxy termini. Most of the abnormalities identified in GS were missense mutations, distributed throughout the TSC gene without a hot spot. A 42-year-old Japanese man was introduced for close examination of hypokalemia. In renal clearance studies using furosemide or thiazide, chloride clearance was increased after furosemide but not after thiazide administration. Furthermore, the distal fractional chloride reabsorption was dramatically decreased by furosemide but not thiazide administration, suggesting a defect in the distal tubule. We then analyzed the TSC gene to confirm the diagnosis of GS, and identified a novel G to T mutation at the acceptor splice site preceding exon 14, resulting in disruption of a conventional 3'AG consensus splice site. Abnormal splicing by this mutation is predicted to cause the formation of truncated TSC with a partial deletion of the transmembrane domain, which will loose the function of transporter. In conclusion, we have identified a unique novel splice site mutation of the TSC gene in GS. The predicted structure of this mutant TSC can conceivably cause an impairment of the transporter activity and thereby be responsible for the development of GS in our patient.


Subject(s)
Bartter Syndrome/genetics , Benzothiadiazines , Mutation, Missense/genetics , RNA Splice Sites/genetics , Sodium Chloride Symporter Inhibitors/pharmacology , Sodium-Potassium-Chloride Symporters/drug effects , Sodium-Potassium-Chloride Symporters/genetics , Adult , Diuretics , Humans , Kidney Tubules, Distal/chemistry , Male , Polymorphism, Restriction Fragment Length , Sodium-Potassium-Chloride Symporters/analysis
3.
Growth Horm IGF Res ; 13(5): 275-81, 2003 Oct.
Article in English | MEDLINE | ID: mdl-12932749

ABSTRACT

We investigated the bioactivity of GH and compared with their immunoactivity in GH bioassay system using lactogenic hormone responsive element (LHRE) reporter gene in Chinese hamster ovary cells transiently co-transfected with human GH receptor cDNA and LHRE/TK-luciferase reporter gene (LHRE/Luc). The recombinant and serum GH but not prolactin almost equally were able to induce LHRE/Luc in a significant and dose-dependent manner, which were equally suppressed by anti-GH. Recombinant GH binding protein (GHBP) at 100 ng/ml but not at 20 ng/ml slightly attenuated GH-induced LHRE/Luc. The serum GH bioactivity (ng/ml) in patients with acromegaly were equal near to their immunoactivity, whereas the bioactivity of the serum GH in a short child with mutant GH (R77C) revealed lower than their immunoactivity. The bioactivity of the recombinant mutant GH was as half as that of wild type GH, thus confirming an antagonistic property of mutant GH. LHRE reporter gene activation assay is useful to measure the GH bioactivity in addition to the conventional bioassay using cell proliferation.


Subject(s)
Biological Assay , Genes, Reporter/genetics , Human Growth Hormone/pharmacology , Prolactin/physiology , Response Elements/genetics , Transcriptional Activation , Animals , CHO Cells , Cell Line , Child , Child, Preschool , Cricetinae , Cricetulus , Female , Human Growth Hormone/blood , Human Growth Hormone/genetics , Humans , Luciferases/genetics , Luciferases/metabolism , Prolactin/metabolism , Receptors, Somatotropin/genetics , Thymidine Kinase/genetics , Transfection , Up-Regulation
4.
Scand J Gastroenterol ; 38(4): 370-9, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12739708

ABSTRACT

BACKGROUND: Helicobacter pylori and duodenogastric reflux (DGR) are both recognized as aetiological factors in chronic gastritis and gastric carcinogenesis. In this study, a Mongolian gerbil (MG) model was used to investigate the histopathological changes in the gastric mucosa resulting from DGR and/or H. pylori infection. METHODS: One-hundred-and-eleven 7-week-old, specific-pathogen-free, male MGs were divided into four groups: normal controls, gerbils with surgically induced DGR, and H. pylori-infected gerbils with and without DGR. Gerbils were killed 4, 12 and 26 weeks after DGR surgery, their stomachs removed and sections prepared. Sections were fixed immediately in 20% phosphate-buffered formalin and subjected to haematoxylin and eosin staining, Alcian blue at pH 2.5/periodic acid-Schiff staining, and immunostaining for smooth muscle cells, H. pylori and 5'-bromo-2'-deoxyuridine (BrdU). RESULTS: The gastric mucosa of H. pylori-infected gerbils showed chronic active gastritis irrespective of DGR throughout the experimental period. The gastric mucosa of H. pylori-infected gerbils with DGR demonstrated higher BrdU labelling than in the other groups. CONCLUSIONS: In MGs, DGR and H. pylori infection synergistically increased gastric mucosal cell proliferative activity. DGR and H. pylori infection may be involved synergistically in gastric carcinogenesis by increasing cell proliferative activity.


Subject(s)
Duodenogastric Reflux/complications , Gastric Mucosa/pathology , Gastritis/pathology , Helicobacter Infections/complications , Helicobacter pylori , Animals , Chronic Disease , Disease Models, Animal , Duodenogastric Reflux/surgery , Gastric Mucosa/microbiology , Gastritis/etiology , Gerbillinae , Male
6.
Acta Neurochir (Wien) ; 143(10): 1031-7, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11685610

ABSTRACT

OBJECTIVE: The purpose of this study is to examine the relation between Glasgow Coma Scale (GCS) score and findings on computed tomography (CT) and magnetic resonance (MR) imaging of patients with mild head injury presenting GCS scores between 13 and 15. METHODS: Data were collected from all consecutive patients with mild head injury who were referred to our hospital between July 1 and October 31, 1999. All patients were recommended to undergo CT and MR imaging examinations. Patients younger than 14 years of age were excluded. RESULTS: Ninety patients were recruited into this study. CT scans were obtained in 88 patients and MR imaging were obtained in 65 patients. Of these 90 patients, 2 patients scored 13 points, 5 scored 14 points and 83 (92.2%) 15 points. Patients with GCS score of 13 points demonstrated parenchymal lesions on both CT and MR imaging. Those with 14 points revealed absence of parenchymal abnormality on CT, but presence of parenchymal lesions on MR imaging. Patients in advanced age (chi square test, p<0.0001), and those with amnesia (p=0005, not significant), although scoring 15 points, revealed a tendency to abnormal intracranial lesions on CT scans. CONCLUSION: It is doubtful whether patients with GCS score 13 should be included in the mild head injury category, due to obvious brain damage on CT scans. MR imaging should be performed on patients with GCS score 14, since the parenchymal lesions are not clearly demonstrated on CT scans. Even if patients scored GCS 15, patients with amnesia or of advanced age should undergo CT scans at minimum, and MR imaging when available.


Subject(s)
Craniocerebral Trauma/classification , Craniocerebral Trauma/pathology , Glasgow Coma Scale , Magnetic Resonance Imaging , Tomography, X-Ray Computed , Adolescent , Adult , Aged , Aged, 80 and over , Amnesia/etiology , Female , Humans , Male , Middle Aged , Prognosis , Prospective Studies
7.
Clin Endocrinol (Oxf) ; 55(2): 175-81, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11531923

ABSTRACT

OBJECTIVE: Skeletal involvement is a common clinical feature in acromegalic patients. Although several recent reports are available concerning bone mineral density (BMD) in acromegaly, the controversy still exists as to whether BMD of acromegalic patients is increased or not. The present study was performed to examine biochemical bone metabolic indices and BMD as well as body composition in 26 Japanese patients with active acromegaly and 26 control subjects matched for age, sex, race and height in a cross-sectional study. MEASUREMENTS: BMD of the lumbar spine and femoral neck, as well as body composition, was measured by dual-energy X-ray absorptiometry. Mid-radial BMD was measured by single-photon absorptiometry. We also determined serum levels of IGF-I, IGFBP-3 and osteocalcin (OC) as well as urinary levels of deoxy-pyridinoline (D-Pyr) and CrossLaps. RESULTS: Percent lean body mass was increased and percent fat mass was decreased in the acromegalic patients compared to control subjects. Serum levels of OC, as well as urinary levels of D-Pyr and CrossLaps, were significantly higher in acromegalic patients compared to control subjects (9.8 +/- 1.2 vs. 5.7 +/- 0.77 for OC; 11.8 +/- 1.66 vs. 5.0 +/- 0.49 for D-Pyr; 437.6 +/- 68.4 vs. 156.5 +/- 39.6 for CrossLaps). Z scores of BMD at mid-radius as well as lumbar spine and femoral neck were significantly higher in acromegalic patients compared to control subjects (1.086 +/- 0.311 vs. -0.060 +/- 0.274 for mid-radius; 1.022 +/- 0.280 vs. 0.319 +/- 0.165 for lumbar spine; 1.292 +/- 0.347 vs. 0.232 +/- 0.264 for femoral neck). CONCLUSIONS: The present study revealed that a decrease in percent fat mass and an increase in percent lean body mass were observed in Japanese patients with active acromegaly. Bone mineral density at all sites and bone metabolic markers were also increased in acromegaly. The present findings provide additional evidence that the GH/IGF-I axis might play an important role in the maintenance of bone mass as well as the regulation of body composition in Japanese adults.


Subject(s)
Acromegaly/metabolism , Body Composition , Bone and Bones/metabolism , Absorptiometry, Photon , Amino Acids/urine , Bone Density , Case-Control Studies , Collagen/urine , Cross-Sectional Studies , Female , Humans , Insulin-Like Growth Factor Binding Protein 3/blood , Insulin-Like Growth Factor I/analysis , Male , Middle Aged , Osteocalcin/blood , Peptide Fragments/urine
8.
J Clin Neurosci ; 8(2): 120-3, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11484659

ABSTRACT

Retrospective analysis of 272 patients with severe head injury was performed. Patient age, Glasgow Coma Scale (GCS) score, pupillary abnormalities, impaired oculocephalic response, presence of subarachnoid haemorrhage, and multiplicity of parenchymal lesions on computerised tomography (CT) were examined. The CT findings were divided into 2 groups, diffuse brain injury (DBI) and mass lesion, according to the classification of the Traumatic Coma Data Bank. The DBI, basically, has no high or mixed density lesion more than 25 ml on CT, and was classified into 4 subgroups: DBI I includes injuries where there is no visible pathology; DBI II includes all injuries in which the cisterns are present with a midline shift of less than 5 mm; DBI III includes injuries with swelling where the cisterns are compressed or absent and the midline shift is less than 5 mm; DBI IV includes injuries with a midline shift of more than 5 mm. The mass lesions were categorised into 3 subgroups: epidural haematoma; acute subdural haematoma; and intracerebral haematoma. Outcomes were determined at 6 months following trauma using the Glasgow Outcome Scale. All DBI I patients recovered well. In the DBI II group, age, GCS score and detection of multiple parenchymal lesions on CT were significantly correlated with outcome. For the DBI III and IV groups, the only significant prognostic factor was the GCS score. In patients with a mass lesion, the GCS score was the only significant prognostic factor in the epidural haematoma group, but the GCS score and the presence of subarachnoid haemorrhage were predictive factors in the acute subdural haematoma group. Outcomes were unfavourable in the majority of patients with intracerebral haematoma. GCS score could predict outcome in all groups. The confidence of the outcome prediction ranged from 75.8 to 92.1%, depending on logistic regression analysis.


Subject(s)
Craniocerebral Trauma/diagnosis , Adolescent , Adult , Child , Child, Preschool , Glasgow Coma Scale , Hematoma, Epidural, Cranial/diagnosis , Hematoma, Subdural/diagnosis , Humans , Intracranial Hemorrhage, Traumatic/diagnosis , Middle Aged , Predictive Value of Tests , Prognosis , Retrospective Studies , Tomography, X-Ray Computed , Treatment Outcome
9.
Life Sci ; 68(26): 2917-23, 2001 May 18.
Article in English | MEDLINE | ID: mdl-11411791

ABSTRACT

A defect in the klotho gene expression in mice leads to a syndrome resembling human aging. The klotho gene encodes a single membrane protein whose extracellular domain carries homology to beta-glucosidases. However, either its function or regulatory mechanism of the gene expression still remains unknown. In the present study, we investigated the klotho gene expression in 3T3-L1 adipocytes using quantitative reverse transcription-polymerase chain reaction. Both membrane and secreted forms of the klotho gene were expressed in 3T3-L1 preadipocytes. Accompanied with adipose differentiation, not the secreted form but the membrane form was gradually increased. In 3T3-L1 adipocytes, triiodothyronine significantly increased the expression levels of membrane form of the klotho gene. These results suggest that the expression of membrane and secreted forms of klotho transcripts are regulated by different mechanisms and that the klotho gene product may play a role in adipose differentiation.


Subject(s)
Adipocytes/physiology , Gene Expression Regulation/physiology , Membrane Proteins/genetics , Triiodothyronine/pharmacology , 3T3 Cells , Adipocytes/cytology , Adipocytes/drug effects , Aging , Animals , Cell Differentiation/physiology , Cell Membrane/metabolism , Dehydroepiandrosterone/pharmacology , Dexamethasone/pharmacology , Gene Expression Regulation/drug effects , Glucuronidase , Humans , Kinetics , Klotho Proteins , Mice , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , Time Factors , Transcription, Genetic
10.
Biochem Biophys Res Commun ; 284(3): 660-6, 2001 Jun 15.
Article in English | MEDLINE | ID: mdl-11396952

ABSTRACT

The aim of this study is to clarify the hormonal regulation of the human ghrelin receptor gene expression in GH(3) cells transfected with our previously cloned 5'-flanking region inserted into a luciferase reporter vector. Phorbor 12-tetradecanoate 13-acetate (TPA) with simultaneous addition of Bay K8644 mimicking ghrelin action caused a significant inhibition of the luciferase activity through the ghrelin receptor gene upstream proximal to -669 but not to -608 base pairs (bp). Glucocorticoid caused a weak but significant inhibition of the luciferase activity through the ghrelin receptor gene upstream proximal to -531 but not to -475 bp. Electrophoretic mobility shift assay resulted in binding of oligonucleotides between -669 and -640 bp, and between -520 and -491 bp to GH(3) cell nuclear proteins unlike AP(2) or glucocorticoid receptor. These results suggest that both TPA/Bay K8644 and glucocorticoid downregulate human ghrelin receptor gene expression through the transcriptional mechanism involving some nuclear factors.


Subject(s)
Receptors, Cell Surface/genetics , Receptors, G-Protein-Coupled , 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester/pharmacology , CREB-Binding Protein , DNA/metabolism , DNA-Binding Proteins/metabolism , Down-Regulation , Humans , Hydrocortisone/pharmacology , Luciferases/genetics , Luciferases/metabolism , Nuclear Proteins/genetics , Pituitary Gland/metabolism , Receptors, Ghrelin , Regulatory Sequences, Nucleic Acid , Tetradecanoylphorbol Acetate/pharmacology , Trans-Activators/genetics , Transcription, Genetic/drug effects , Transfection , Tumor Cells, Cultured
11.
J Biol Chem ; 276(33): 31422-8, 2001 Aug 17.
Article in English | MEDLINE | ID: mdl-11390373

ABSTRACT

Exogenous administration of eicosapentaenoic acid (EPA) improves insulin sensitivity, but its precise mechanism remains unknown. Here we show that EPA stimulates the intracellular insulin signaling pathway in hepatoma cells. Exposure of these cells to EPA caused up-regulation of several insulin-induced activities including tyrosine phosphorylation of insulin receptor substrate-1, insulin receptor substrate-1-associated phosphatidylinositol 3-kinase, and its downstream target Akt kinase activity as well as down-regulation of gluconeogenesis. In contrast, EPA decreased mitogen-activated protein kinase activity and inhibited cell proliferation. These findings raise the possibility that EPA up-regulates metabolic action of insulin and inhibits cell growth in humans.


Subject(s)
Adaptor Proteins, Signal Transducing , Eicosapentaenoic Acid/pharmacology , Insulin/pharmacology , Carcinoma, Hepatocellular/pathology , Cell Division , GRB2 Adaptor Protein , Glucosamine/metabolism , Glucose/metabolism , Humans , Insulin Receptor Substrate Proteins , Mitogen-Activated Protein Kinases/metabolism , Phosphatidylinositol 3-Kinases/physiology , Phosphoproteins/physiology , Phosphorylation , Proteins/physiology , Receptors, Cytoplasmic and Nuclear/physiology , Transcription Factors/physiology , Tumor Cells, Cultured , Tyrosine/metabolism , Up-Regulation
12.
J Clin Microbiol ; 39(4): 1283-8, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11283042

ABSTRACT

The Mongolian gerbil model for Helicobacter pylori infection is an animal model that mimics human disease. We examined the serum immune response to H. pylori infection in gerbils by enzyme-linked immunosorbent assay (ELISA) and Western blotting, both with whole-cell (H. pylori) extracts. A total of 66 7-week-old specific-pathogen-free male gerbils were inoculated orogastrically with H. pylori strain ATCC 43504. Sera were collected 1, 2, 4, 8, 12, 26, 38, and 52 weeks after H. pylori inoculation. Sixty-nine noninfected gerbils and their sera were used as controls. The specificity of the ELISA was 95.7%. The frequency of seropositivity increased over time: 2 of 10 (20%), 7 of 10 (70%), and 7 of 7 (100%) samples of sera from inoculated gerbils were positive for H. pylori at 2, 4, and 8 weeks postinoculation, respectively. Western blot assays showed that the primary immunoglobulin G (IgG) response against low-molecular-mass (25-, 30-, and 20-kDa) proteins appeared after a lag period of 2 to 8 weeks after inoculation. Antibodies against 160-, 150-, 110-, 120-, 80-, 66-, and 63-kDa proteins were observed 12 weeks after inoculation. The early reactive 30-kDa protein was identified as a urease alpha subunit by N-terminal amino acid sequencing. After 26 weeks, two groups of animals could be distinguished: one group developed ulcers (n = 5), and the other developed hyperplastic polyps without ulcers (n = 19). Gerbils in the gastric ulcer group showed significantly higher serum anti-H. pylori IgG levels than did gerbils in the hyperplastic group (P = 0.001) as measured by ELISA. Furthermore, a higher proportion of animals developed antibodies to H. pylori proteins of 26, 25, and 20 kDa in the ulcer group than those animals with hyperplastic polyps (75 to 100% versus 17 to 50%) in Western blot assays. These results highlight the importance of the immune response of the host in the development of H. pylori-related gastric lesions.


Subject(s)
Antibodies, Bacterial/blood , Antigens, Bacterial/immunology , Helicobacter Infections/immunology , Helicobacter pylori/immunology , Immunoglobulin G/blood , Animals , Blotting, Western , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Gerbillinae , Helicobacter Infections/microbiology , Humans , Male
13.
Biochem Biophys Res Commun ; 276(2): 411-6, 2000 Sep 24.
Article in English | MEDLINE | ID: mdl-11027489

ABSTRACT

Recently a novel peptide which specifically stimulates the secretion of prolactin (PRL) was found and named PRL-releasing peptide (PrRP). To evaluate the regulation of human (h) PrRP-receptor (PrRP-R) gene expression, we cloned the 5'-flanking region of the hPrRP-R gene and determined the nucleotide sequence of 4.0 kilobase pairs (kb) upstream from the translation start site. Analysis of the hPrRP-R transcripts by means of 5'-rapid amplification of cDNA ends suggested that the hPrRP-R gene had multiple transcription start sites between -429 and -365 from the translation start site. There is no typical TATA or CAAT but a GC box and putative binding sites for several transcription factors including Pit-1 and pituitary homeobox 1 (Ptx1). However, transient transfection studies using a luciferase reporter gene demonstrated that 5'-flanking region exerts promoter activity in several non-pituitary cell lines as well as in GH(3) cells. The GC box located from -467 to -457 was identified as an important region for the basal expression of the hPrRP-R gene. Knowledge of the promoter region of the hPrRP-R gene, which was obtained in the present study, will facilitate the clarification of its transcriptional regulation.


Subject(s)
Gene Expression Regulation , Promoter Regions, Genetic/genetics , Receptors, Neuropeptide/genetics , Animals , Base Sequence , Cells, Cultured , Cloning, Molecular , DNA/analysis , Humans , Molecular Sequence Data , Rats
14.
Endocr J ; 47(3): 335-42, 2000 Jun.
Article in English | MEDLINE | ID: mdl-11036878

ABSTRACT

Multifocal fibrosclerosis denotes a combination of similar fibrous disorders occurring at different anatomical sites. We encountered a 53-year-old male patient with orbital pseudotumor, chronic paranasal sinusitis, fibrous nodules of the lungs, intracranial pachymeningitis, and panhypopituitarism with central diabetes insipidus (DI) as a possible manifestation of multifocal fibrosclerosis. It has been reported that intracranial pachymeningitis or orbital pseudotumor associated with multifocal fibrosclerosis could invade the sella turcica causing a variety of anterior and/or posterior pituitary dysfunctions. In our case, intracranial pachymeningitis apparently involved the pituitary stalk and gland. Isolated gonadotropin deficiency, in addition to central DI, preceded panhypopituitarism. Although panhypopituitarism with central DI due to multifocal fibrosclerosis is quite rare and only one case has ever been reported, this systemic fibrotic disorder can be a possible cause of panhypopituitarism with central DI.


Subject(s)
Diabetes Insipidus/complications , Hypopituitarism/etiology , Fibrosis , Follicle Stimulating Hormone/blood , Headache/complications , Humans , Luteinizing Hormone/blood , Male , Middle Aged , Sclerosis , Sinusitis/complications
15.
Article in Japanese | MEDLINE | ID: mdl-11004708

ABSTRACT

We evaluated Helicobacter Selective Agar (HSA) medium (Nissui Pharmaceuticals Co., Ltd., Tokyo, Japan), which has been newly developed and has been commercially available since June in 1998. HSA medium strongly inhibited the growth of possible contaminants from gastric biopsy-specimens, such as alpha-streptococci, Neisseria species, Enterococcus gallinarum, Pseudomonas aeruginosa, Corynebacterium species, Capnocytophaga species, and Candida albicans. Cultures run in parallel with Helicobacter Pylori Agar (HPA) medium (Eiken Chemical Co., Ltd., Tokyo, Japan) disclosed no discordance in the detective rates of H. pylori strains between the media examined. Inaddition, the numbers of the colonies and the colony-sizes grown on the plates were also in good agreement with each other. However, it would be strikingly favorable that the appearing colonies on the HSA medium turned purple. This coloration enabled us to detect the emergence of the colonies much earlier and easier when compared with the conventional other media including HPA medium (Eiken). These findings led us to conclude that HSA medium (Nissui) was superior to HPA medium (Eiken) in the prompt detection and the rapid identification of H. pylori in routine clinical microbiology.


Subject(s)
Culture Media , Helicobacter pylori/isolation & purification , Agar , Bacteriological Techniques , Humans , Stomach/microbiology
16.
J Clin Microbiol ; 38(6): 2409-11, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10835018

ABSTRACT

The susceptibilities of 20 strains of vancomycin-resistant enterococci (VRE) with the vanB genotype obtained by using Vitek GPS-418 cards were compared with those obtained by the broth dilution method of the National Committee for Clinical Laboratory Standards (NCCLS) (approved standard M7-A4) and with those obtained by the agar screen method using bile esculin azide agar containing 6 microgram of vancomycin per ml. Although both the broth dilution and agar screen methods disclosed no discordance, Vitek GPS-418 cards yielded a very major error compared with the results obtained by the reference broth dilution method of the NCCLS. Vitek GPS-418 cards were therefore found to have considerable room for improvement for the accurate detection of vanB VRE strains.


Subject(s)
Bacterial Proteins/genetics , Enterococcus faecalis/drug effects , Microbial Sensitivity Tests/instrumentation , Vancomycin Resistance/genetics , Enterococcus faecalis/genetics , Gram-Positive Bacterial Infections/microbiology , Humans , Reproducibility of Results
17.
Antimicrob Agents Chemother ; 44(2): 458-61, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10639386

ABSTRACT

The MICs of rabeprazole sodium (RPZ), a newly developed benzimidazole proton pump inhibitor (PPI), against 133 clinical Helicobacter pylori strains revealed a higher degree of activity than the another two PPIs, lansoprazole and omeprazole. Time-kill curve assays of RPZ, when combined with amoxicillin, clarithromycin, or metronidazole, disclosed that synergistic effects were demonstrated in combination with each antibiotic examined. Moreover, no apparent antagonistic effect appeared among all of the strains tested.


Subject(s)
Anti-Ulcer Agents/pharmacology , Benzimidazoles/pharmacology , Helicobacter pylori/drug effects , Proton Pump Inhibitors , 2-Pyridinylmethylsulfinylbenzimidazoles , Anti-Bacterial Agents/pharmacology , Benzimidazoles/chemistry , Drug Interactions , Humans , Microbial Sensitivity Tests , Omeprazole/analogs & derivatives , Rabeprazole
18.
Biochem Biophys Res Commun ; 270(2): 343-8, 2000 Apr 13.
Article in English | MEDLINE | ID: mdl-10753628

ABSTRACT

Recent evidence indicates that both leptin and eicosapentaenoic acids (EPA) improve insulin sensitivity. In the present study, we examined the effect of EPA on endogenous leptin expression in 3T3-L1 adipocytes to clarify whether the EPA's effect is exerted through leptin expression. EPA caused a time- and dose-dependent increase of leptin mRNA levels in 3T3-L1 adipocytes. Leptin mRNA expression was significantly increased up to 309.4 +/- 17.0% of the control by 24 h (P < 0.01; n = 6). Leptin secretion was also significantly increased up to 193.3 +/- 12.1% of the control by 24 h (P < 0.01; n = 6). EPA is a ligand for peroxisome proliferator-activated receptors (PPARs) with the highest affinity to PPARalpha. We examined the effect on leptin expression of clofibrate, a ligand for PPARalpha, bezafibrate, for PPARbeta, or troglitazone, for PPARgamma, to clarify whether these ligands for PPARs could mimic EPA-induced stimulation of leptin expression. Neither clofibrate nor bezafibrate affected leptin mRNA expression, whereas troglitazone significantly suppressed leptin mRNA expression. On the other hand, inhibition by 6-diazo-5-oxo-l-norleucine of the rate-limiting enzyme in hexosamine biosynthesis blunted EPA-induced stimulation of leptin mRNA expression and its secretion. These data suggest that EPA up-regulates leptin gene expression and its secretion probably through a hexosamine biosynthetic pathway.


Subject(s)
Adipocytes/drug effects , Gene Expression Regulation/drug effects , Leptin/genetics , RNA, Messenger/genetics , 3T3 Cells , Adipocytes/metabolism , Animals , Base Sequence , DNA Primers , Diazooxonorleucine/pharmacology , Leptin/metabolism , Mice , Molecular Sequence Data , Receptors, Cytoplasmic and Nuclear/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Transcription Factors/metabolism
20.
J Clin Endocrinol Metab ; 84(11): 4214-9, 1999 Nov.
Article in English | MEDLINE | ID: mdl-10566675

ABSTRACT

A missense mutation, C422F, was identified in the intracellular domain of GH receptor (GHR) in a Japanese short boy. Although this mutation was previously reported in a patient with GH insensitivity syndrome (GHIS), it has not been clear whether this mutation causes GH insensitivity. To clarify the effect of this mutation on GH signal transduction, mutant GHR was expressed in CHO cells, and its functional properties were investigated. There were no significant differences in GH-induced tyrosine phosphorylation of STAT5b (signal transducer and activator of transcription) between wild-type GHR (GHR-wt)- and mutant GHR (GHR-C422F)-expressing cells. Moreover, STAT5-mediated transcriptional activation of GHR-C422F-expressing cells was comparable to that of GHR-wt-expressing cells. These findings indicated that the C422F mutation of GHR affected neither GH-induced tyrosine phosphorylation nor the transcriptional activation of STAT5. In addition, the analysis of genotypes and phenotypes of his family revealed that body heights of family members with heterozygous or homozygous C422F mutations were all within normal ranges, with the single exception of the proband. These in vitro and in vivo results indicate that the C422F missense mutation of GHR is a polymorphism that does not result in GHIS.


Subject(s)
Body Height , Milk Proteins , Mutation , Receptors, Somatotropin/genetics , Animals , CHO Cells , Child, Preschool , Cricetinae , DNA-Binding Proteins/metabolism , Gene Expression , Genotype , Heterozygote , Human Growth Hormone/pharmacology , Humans , Male , Pedigree , Phosphorylation , Phosphotyrosine/metabolism , STAT5 Transcription Factor , Trans-Activators/metabolism , Transcription, Genetic
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