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T Ohkubo

Publications and source records attributed to T Ohkubo.

At least 91 records · Page 5Linked to original sources
PubMed2000

[Two cases of acute lupus peritonitis].

We report two cases of systemic lupus erythematosus (SLE) diagnosed when acute peritonitis was appeared. Case 1 was a 20 year-old woman suffering from stomachache and right lower abdominal pain. Case 2 was a 40 year-old woman with diarrhea, epigastralgia, pollakisuria. In both cases, their peritoneal fluids were exudative with positive autoantibodies. After high dose steroid therapy, abdominal symptoms and ascites improved promptly. However, due to the complication of lupus nephritis, additional therapy was necessary. To characterize the feature of lupus peritonitis (LP), we examined the clinical and laboratory findings of LP from the literature. In patients with acute LP, abdominal pain, vomiting, diarrhea were significantly more common compared with chronic LP patients (P < 0.05), and fever, arthritis, central nervous system involvement and cystitis were more common. In patients with chronic LP, pleural effusion and pericardial effusion were more common compared with acute LP patients. Gastrointestinal manifestations such as abdominal pain, vomiting and diarrhea were more common in patients with acute LP compared with patients with chronic LP. Most patients with chronic LP were asymptomatic, ascites and serositis being the only clinical findings. The response to steroid therapy was better in acute LP.

Acute Disease↗
PubMed1999

Enhancement of lipocalin-type prostaglandin D synthase enzyme activity by guanidine hydrochloride.

The characterization of unfolding of mouse recombinant lipocalin-type prostaglandin D synthase (L-PGDS) by guanidine hydrochloride (GdnHCl) was carried out. In the presence of low concentrations of GdnHCl (up to 0.75 M), enhancement of the enzyme activity was observed. However, above a 1 M concentration of GdnHCl, the enzyme activity was reduced in a concentration-dependent manner. The maximum enzyme activity induced by GdnHCl was approximately 1. 5-fold compared with the activity under physiological conditions without GdnHCl. The ellipticity in circular dichroism (CD) spectrum of the L-PGDS at 218 nm, reflecting the beta-sheet content, was decreased by GdnHCl (up to 0.75 M), and the minimum ellipticity was observed at 0.5 M GdnHCl. The fluorescence quenching of the intrinsic tryptophan of L-PGDS due to the binding of bilirubin in the presence or absence of GdnHCl was measured. The K(d) values obtained in the presence and absence of 0.5 M GdnHCl were 447 and 115 nM, respectively, indicating lower affinity of the L-PGDS for bilirubin with GdnHCl than without it. Further, an NMR study revealed that the reorganization of hydrogen-bond network in the L-PGDS was observed in the presence of 0.5 M GdnHCl. These results, taken together, indicate that the enzyme activity of L-PGDS is enhanced by the conformational change, especially by the change in the secondary structure.

Animals↗
PubMed1999

Mode analysis of a fatty acid molecule binding to the N-terminal 8-kDa domain of DNA polymerase beta. A 1:1 complex and binding surface.

We reported previously that long-chain fatty acids are potent inhibitors of mammalian DNA polymerase beta. At present, based on information available from the NMR structure of the N-terminal 8-kDa domain, we examined the structural interaction with the 8-kDa domain using two species, C(18)-linoleic acid (LA) or C(24)-nervonic acid (NA). In the 8-kDa domain with LA or NA, the structure that forms the interaction interface included helix-1, helix-2, helix-4, the three turns (residues 1-13, 48-51, and 79-87) and residues adjacent to an Omega-type loop connecting helix-1 and helix-2 of the same face. No significant shifts were observed for any of the residues on the opposite side of the 8-kDa domain. The NA interaction interface on the amino acid residues of the 8-kDa domain fragment was mostly the same as that of LA, except that the shifted cross-peaks of Leu-11 and Thr-79 were significantly changed between LA and NA. The 8-kDa domain bound to LA or NA as a 1:1 complex with a dissociation constant (K(D)) of 1.02 or 2.64 mM, respectively.

Animals↗
PubMed1999

p120(ctn) binds to the membrane-proximal region of the E-cadherin cytoplasmic domain and is involved in modulation of adhesion activity.

Cadherins are transmembrane glycoproteins involved in Ca(2+)-dependent cell-cell adhesion. Previously, we showed that the conserved membrane-proximal region of the E-cadherin cytoplasmic domain negatively regulates adhesion activity. In this report, we provide several lines of evidence that p120(ctn) is involved in this negative regulation. p120(ctn) binds to the membrane-proximal region of the nonfunctional carboxyl-terminally deleted E-cadherin protein. An additional internal deletion in this region prevented the association with p120(ctn) and activated the protein, as seen in an aggregation assay. Furthermore, the nonfunctional E-cadherin can be activated through coexpression of p120(ctn) proteins with amino-terminal deletions, which eliminate several potential serine/threonine phosphorylation sites but do not affect the ability to bind to cadherins. Finally, we show that staurosporine, a kinase inhibitor, induces an increased electrophoretic mobility of p120(ctn) bound to E-cadherin polypeptides, activates the nonfunctional E-cadherin protein, and converts the wild-type E-cadherin and an E-cadherin-alpha-catenin chimeric protein from a cytochalasin D-sensitive to a cytochalasin D-insensitive state. Together, these results indicate that p120(ctn) is a modulator of E-cadherin-mediated cell adhesion.

Animals↗
PubMed1999

A novel splicing mutation (894-9 G --> A) of the MEN1 gene responsible for multiple endocrine neoplasia type 1.

Germline MEN1 gene mutations are responsible for multiple endocrine neoplasia type 1 (MEN1), a dominantly inherited cancer syndrome. We identified a MEN1 germline mutation 894-9 G --> A in three MEN1 patients from two unrelated families. This mutation was not present in any of the 100 blood samples from normal volunteers. The wild type MEN1 sequence was lost in the patient's pancreatic tumor. Abnormal mRNA was identified in the tumor, which retained an intronic sequence indicating aberrant mRNA splicing at a newly created splicing acceptor site. These findings indicate that this nucleotide substitution is, though previously reported to be a polymorphism, a causative splicing mutation.

Adult↗
PubMed1999

Solution structure of an insect growth factor, growth-blocking peptide.

Growth-blocking peptide (GBP) is an insect growth factor consisting of 25 amino acid residues that retards the development of lepidopteran larvae at high concentration while it stimulates larval growth at low concentration. In this study, we determined the solution structure of GBP by two-dimensional 1H NMR spectroscopy. The structure contains a short segment of double-stranded beta-sheet involving residues 11-13 and 19-21 and a type-II beta-turn in the loop region (residues 8-11), whereas the N and C termini are disordered. This is the first report of the three-dimensional structure of the peptiderigic insect growth factor, and the structure of the well defined region of GBP was found to share similarity with that of the C-terminal domain of the epidermal growth factor (EGF). Because GBP has been reported to stimulate DNA synthesis of not only insect cells but also human keratinocyte cells at the same level with EGF, the structural similarity between GBP and EGF may lead to the interaction of GBP to EGF receptor.

Amino Acid Sequence↗
PubMed1999

Monitoring of zonisamide in human breast milk and maternal plasma by solid-phase extraction HPLC method.

An HPLC method was developed for the determination of zonisamide in human breast milk and plasma. Chromatographic separation was achieved using a Develosil CN analytical column with potassium dihydrogenphosphate buffer (pH 3.5 with milk, pH 2.5 with plasma)-acetonitrile as the mobile phase. Zonisamide and 1, 2-benzisoxazole-3-methansulfonamine acetate as internal standard were detected by ultraviolet absorbance at 240 nm. Zonisamide in breast milk and plasma was extracted by a rapid and simple procedure based on C(18) bonded-phase extraction. Determination of zonisamide in human breast milk and plasma was possible in the concentration range 0.05-20.0 microg/mL. The recoveries of zonisamide added to human breast milk and plasma were 79.5-85.0% and 86.3-93.1%, respectively, with coefficients of variation of less than 8.3% and 11.4% respectively. The mean concentrations of zonisamide in breast milk and plasma were 9.41 +/- 0.95 and 10.13 +/- 0.45 microg/mL, respectively. The average ratio between the breast milk concentration and plasma concentration (M/P ratio) was 0.93 +/- 0.09.

Adult↗
PubMed1999

cDNA cloning and developmental alterations in gene expression of the two Pit-1/GHF-1 transcription factors in the chicken pituitary.

Pit-1/GHF-1 (Pit-1) transcription factors promote the gene expressions for growth hormone (GH), prolactin (PRL), and the beta chain of thyroid-stimulating hormone in vertebrate pituitary glands. The present study analyzed the nature of chicken Pit-1s (cPit-1s) and their developmental expressions in the pituitary. Chicken pituitary expressed two cPit-1 mRNAs encoding cPit-1alpha and cPit-1gamma composed of 335 and 327 amino acid residues, respectively. They possessed different N-terminal regions and the common C-terminal regions containing a POU-specific domain and a POU homeodomain. Northern blot analysis revealed the pituitary-specific expressions of these Pit-1 mRNAs, and the Pit-1alpha mRNA expressions were two to three times higher than those for Pit-1gamma in both cephalic and caudal lobes of the pituitary. The cPit-1alpha and gamma mRNA expressions simultaneously increased after hatching until 4 weeks and then slightly decreased at 5 weeks. Similar gene expression profiles were observed for GH and PRL during the posthatch developmental period.

Amino Acid Sequence↗
PubMed1999

Transabdominal oxygenation using perfluorocarbons.

PURPOSE: Evaluation of the intraabdominal (intraperitoneal and intraluminal) administration of oxygen-saturated perfluorocarbon on both portal and arterial blood oxygenation. METHODS: Eight male rabbits were divided into the test (n = 5) and control (n = 3) groups. Each underwent intrajejunal, intraperitoneal, and intravascular (artery, portal vein) catheter placements along with ligation of the duodenum and the terminal ileum under general anesthesia. The test group received oxygen-saturated perfluorotripropylamine (FTPA), and the control group received oxygen desaturated FTPA. The oxygen delivery was assessed by serial blood gas measurements before and after the administration of FTPA. RESULTS: The administration of oxygen-saturated FTPA significantly increased the partial pressure of oxygen within both the arterial and the portal venous blood (PaO2, PpVO2) without significant changes in PCO2 values. Oxygen desaturated FTPA failed to show any effects on blood gas values. Compared with oxygen desaturated FTPA, oxygen-saturated FTPA increased PaO2, PpVO2, and oxygen saturation (artery, portal vein) significantly at some, but not all of the time-points measured. CONCLUSIONS: The intraabdominal administration of saturated FTPA improved both the portal venous and the arterial oxygenation. This new mode of oxygenation may be helpful as an adjunct to conventional oxygen delivery systems.

Animals↗
PubMed1999

Role of CD95/Fas and its ligand in the regulation of the growth of human CD34(++)CD38(-) fetal liver cells.

The functional significance of CD95/Fas expressed by candidate hematopoietic stem cells (HSCs) from human fetal liver was studied by testing the effect of agonistic anti-CD95 monoclonal antibody (mAb) CH-11 and soluble CD95 ligand (sCD95L) on the growth of CD34(++)CD38(-)lineage cells in vitro. Candidate fetal HSCs exhibited a dose-dependent proliferative response to CH-11 as well as to sCD95L when combined with kit ligand (KL) + interleukin 3 (IL-3) under serum-deprived culture conditions. CH-11 mAb increased, in a synergistic fashion, the number of myeloid colony-forming unit culture (CFU-C) generated by candidate HSCs in liquid cultures with the cytokine combinations KL + IL-3, KL + granulocytemacrophage colony-stimulating factor, and KL + IL-6. CH-11 mAb and sCD95L also enhanced erythropoiesis supported by KL + IL-3 + erythropoietin (Epo). Furthermore, sCD95L was able to increase the number of megakaryocytes, granulocytes, and CD34- cells generated in the presence of KL + IL-3 + Epo + thrombopoietin. An analysis performed using Western blotting revealed that the membrane-bound CD95L (mCD95L) was expressed by both immature (total CD34+/++) and mature (CD34-) hematopoietic lin(-) FL cells. Among the CD34(++)lin(-)cells, both the freshly isolated CD38+ and CD38 subsets as well as CD95+ and CD95- cells constitutively expressed mCD95L, demonstrating that the CD95/CD95L system represents a paracrine and potentially autocrine regulator of early hematopoiesis. To study the role of the endogenously produced CD95L, we determined the effects of a neutralizing anti-CD95L NOK-1 on the growth of candidate HSCs. By blocking the endogenous CD95L with NOK-1 mAb, we observed an increase in CFU-C generated by candidate HSCs. We conclude that the endogenous CD95L has an inhibitory effect on fetal candidate HSCs, which can be blocked by sCD95L and CH-11 mAb.

ADP-ribosyl Cyclase↗
PubMed1999

Determination of risperidone and 9-hydroxyrisperidone in human plasma by high-performance liquid chromatography: application to therapeutic drug monitoring in Japanese patients with schizophrenia.

A high-performance liquid chromatographic method has been developed for the simultaneous determination of risperidone and its major active metabolite 9-hydroxyrisperidone in plasma. Risperidone and 9-hydroxyrisperidone in plasma were extracted using a CN bonded-solid phase cartridge, followed by, C4 reversed-phase HPLC separation. Risperidone, 9-hydroxyrisperidone and trazodone as an internal standard were detected by ultraviolet absorbance at 280 nm. It was possible to determine risperidone in the concentration range of 1.0-100.0 ng/ml(-1) and 9-hydroxyrisperidone at a range of 2.0-200.0 ng/ml(-1). The detection limits of risperidone and 9-hydroxyrisperidone were 0.5 and 1.0 ng/ml(-1), respectively. The mean recoveries of risperidone and 9-hydroxyrisperidone added to plasma were less than 92.0 and 92.6%, with a coefficient of variation of less than 10.6 and 10.5%, respectively. This method has been used for the simultaneous determination of steady-state plasma concentration (Css) of risperidone and 9-hydroxyrisperidone in schizophrenic patients treated with 3-, 6-, and 12-mg risperidone oral doses per day.

Adult↗
PubMed1999

Inter- and intra-individual variation of food and nutrient consumption in a rural Japanese population.

OBJECTIVES: To determine the relative contribution of intra- and inter-individual variation in the consumption of foods and nutrients. DESIGN: Cross-sectional study. SETTING: Two rural towns in the Miyagi Prefecture, a northeastern part of Japan. SUBJECTS: Fifty-nine men and sixty women. METHODS: Four 3-d food records were collected in four seasons within a year. The total variance in the consumption of 15 nutrient variables and 16 food groups was partitioned by analysis of variance into its inter- and intra-individual components, separately for men and women. RESULTS: The ratio of the intra- to inter-individual variance was larger than unity in 87% (13/15) of the nutrients in men and 100% (15/15) in women. The ratio was largest for retinol (52.9 in men and infinity in women) and smallest for carbohydrates (0.8 in men and 1.5 in women). The ratio was larger than unity in 88% (14/16) of the food groups in men and 94% (15/16) in women, which was smallest for cereals (0.6 in men and 0.9 in women). CONCLUSIONS: The results were consistent with previous western studies in that intra-individual variation was generally larger than inter-individual variation for the consumption of both foods and nutrients. SPONSORSHIP: This study was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture (09470106) and by Health Sciences Research Grants for Research on Health Services (H10-25) from the Ministry of Health and Welfare in Japan.

Aged↗
PubMed1999

Peripheral blood neutrophils of germ-free rats modified by in vivo granulocyte-colony-stimulating factor and exposure to natural environment.

Peripheral blood neutrophils from germ-free (GF), specific-pathogen-free (SPF) and conventional (CV) rats were compared. Besides neutropenia and impaired superoxide anion generation as previously reported, it was found that GF rats had lower phagocytic function (70%) and generated less nitric oxide than the other rats. GF and SPF rats were injected with recombinant human granulocyte-colony-stimulating factor (rhG-CSF), or were transferred to natural environment. It was found that total number, phagocytosis, intracellular killing, ratio of phagocytosis versus killing (killing efficiency) and nitric oxide production induced by recombinant rat interferon-gamma (rrIFN-gamma) were normalized upon injection of rhG-CSF. These results indicate that rhG-CSF may stimulate neutrophil production and induce the expression of neutrophil receptors for phagocytosis and nitric oxide production in GF rats. Although lower than in CV rats, the level of superoxide produced was sufficient for normal neutrophil-killing efficiency in SPF and GF rats. In SPF rats, this could be amended by exposure to natural environment. However, neither rhG-CSF injection nor transfer to natural environment could increase the generation of superoxide anion in GF rats.

Animals↗
PubMed1999

A mini-protein designed by removing a module from barnase: molecular modeling and NMR measurements of the conformation.

A globular domain can be decomposed into compact modules consisting of contiguous 10-30 amino acid residues. The correlation between modules and exons observed in different proteins suggests that each module was encoded by an ancestral exon and that modules were combined into globular domains by exon fusion. Barnase is a single domain RNase consisting of 110 amino acid residues and was decomposed into six modules. We designed a mini-protein by removing the second module, M2, from barnase in order to gain an insight into the structural and functional roles of the module. In the molecular modeling of the mini-protein, we evaluated thermodynamic stability and aqueous solubility together with mechanical stability of the model. We chemically synthesized a mini-barnase with (15)N-labeling at 10 residues, whose corresponding residues in barnase are all found in the region around the hydrophobic core. Circular dichroism and NMR measurements revealed that mini-barnase takes a non-random specific conformation that has a similar hydrophobic core structure to that of barnase. This result, that a module could be deleted without altering the structure of core region of barnase, supports the view that modules act as the building blocks of protein design.

Bacterial Proteins↗
PubMed1999

Characteristics of blood pressure measured at home in the morning and in the evening: the Ohasama study.

OBJECTIVE: To determine the qualitative and quantitative differences of blood pressure measured at home (home measurement) in the morning versus the evening. METHODS: Of 3744 participants, aged 20 years or older in the Ohasama population, more than 14 home measurements in the morning and in the evening, respectively, were obtained in each of 1207 individuals (881 untreated, 56.1 +/- 11.4 years and 326 treated, 66.0 +/- 9.2 years). A casual/screening measurement was also obtained in these individuals. RESULTS: The home measurements in the morning were significantly higher than those in the evening. The bivariate linear regression analysis demonstrated that the difference between diastolic home measurement in the morning and that in the evening increased with an increase in diastolic home measurements. The multiple step-wise linear regression analysis, however, demonstrated that male sex, the use of antihypertensive medication, and SD of home measurements in individuals (blood pressure variability), but not level of home measurements, were positively associated with the difference between home measurement in the morning and that in the evening. The SD of home measurement in the evening in individuals was significantly larger than that in the morning, and the SD in treated individuals was significantly larger than that in untreated individuals. The correlations between casual and home measurements were moderate in untreated individuals (r = 0.509-0.567) but poor in treated subjects (r= 0.223-0.384). The correlations between home systolic measurements in the morning and in the evening were very close in both treated and untreated subjects (r = 0.814-0.902). The correlations between the SD of home measurements in the morning and in the evening were moderate in both treated and untreated individuals (r = 0.585-0.657). CONCLUSIONS: Qualitative and quantitative differences in home blood pressure measurement, due to the differential time of measurement, should be taken into consideration in clinical use of home blood pressure measurements.

Adult↗
PubMed1999

Medical cost for disability: a longitudinal observation of national health insurance beneficiaries in Japan.

OBJECTIVES: To measure the impact of disability on the use of medical care and its costs. DESIGN: A 1-year prospective cohort study of National Health Insurance beneficiaries in a rural Japanese community. Their physical function was examined by the Medical Outcomes Study questionnaire at the end of 1994; medical care and its costs were then monitored for 1 year. SETTING AND PARTICIPANTS: Participants were the 49,364 subjects, aged 40 to 79 years, who were beneficiaries of National Health Insurance and lived in the catchment area of Ohsaki Public Health Center, Miyagi, Japan. The subjects were mainly farmers, self-employed persons, housewives, or pensioners. MEASUREMENTS: Medical care utilization (number of outpatient visits and days of inpatient care) and the costs for each subject were obtained from National Health Insurance Claim History files. These measurements were collected from January to December 1995, and the relationship between physical functioning levels and medical costs was analyzed. RESULTS: The medical costs per capita increased with poorer physical function. Medical costs among those with limitations in performing self-care increased by 4 times in men and 3 times in women compared with those with no functional limitation. In this cohort, the 4.3% of the subjects who were dependent in self-care used 15% of the total inpatient days and 10% of the total medical costs. CONCLUSION: Treatment of patients with disability requires a huge amount of medical resources. There is an urgent need for cost-effective intervention programs for disability prevention, which could be offset against the cost for treating the disabled.

Activities of Daily Living↗
PubMed1999

Changes in endothelium-derived vascular regulatory factors during dobutamine-stress-induced silent myocardial ischemia in patients with Kawasaki disease.

The changes in endothelium-derived vascular regulatory factors during dobutamine (DOB)-induced myocardial ischemia (MI) were investigated in 21 patients with Kawasaki disease aged from 11 months to 18 years. They were classified into an ischemia group (8 patients) and a non-ischemia group (13 patients) based on the results of 99mTc myocardial scintigraphy and DOB stress 99mTc myocardial scintigraphy. In the ischemia group, MI was relatively mild, because there were ischemic changes on the electrocardiogram and no significant symptoms during DOB stress. Catheters were positioned near the orifice of the coronary artery (Ao) and at the coronary sinus (CS). Hemodynamics and the blood concentrations of lactic acid and endothelin-1, as well as NO3-, 6-keto-prostaglandin F1alpha, and thromboxane B2, (which are inactive metabolites of nitric oxide, prostaglandin I2 and thromboxane A2, respectively), were measured at rest and after DOB stress (maximum dose: 30 microg x kg(-1) x min(-1)). The CS/Ao ratio was determined for all parameters. The rate-pressure product, an index of work load, and the cardiac index were significantly increased by DOB stress in both groups. Coronary angiography showed no vasospasm of the epicardial coronary arteries before or after DOB stress in either group. The plasma concentrations of endothelin-1 and 6-keto-prostaglandin F1alpha were significantly increased after DOB stress in the ischemia group, but the serum concentration of NO did not increase. The lack of an increase in NO production during DOB stress may have contributed to the worsening of MI in patients with Kawasaki disease.

Adolescent↗