Search PubMed⌕ Search

Biomedical subjects

K Tsuji

Publications and source records attributed to K Tsuji.

At least 55 records · Page 3Linked to original sources
PubMed2001

Enhancement of osteoclastic bone resorption and suppression of osteoblastic bone formation in response to reduced mechanical stress do not occur in the absence of osteopontin.

Reduced mechanical stress to bone in bedridden patients and astronauts leads to bone loss and increase in fracture risk which is one of the major medical and health issues in modern aging society and space medicine. However, no molecule involved in the mechanisms underlying this phenomenon has been identified to date. Osteopontin (OPN) is one of the major noncollagenous proteins in bone matrix, but its function in mediating physical-force effects on bone in vivo has not been known. To investigate the possible requirement for OPN in the transduction of mechanical signaling in bone metabolism in vivo, we examined the effect of unloading on the bones of OPN(-/-) mice using a tail suspension model. In contrast to the tail suspension-induced bone loss in wild-type mice, OPN(-/-) mice did not lose bone. Elevation of urinary deoxypyridinoline levels due to unloading was observed in wild-type but not in OPN(-/-) mice. Analysis of the mechanisms of OPN deficiency-dependent reduction in bone on the cellular basis resulted in two unexpected findings. First, osteoclasts, which were increased by unloading in wild-type mice, were not increased by tail suspension in OPN(-/-) mice. Second, measures of osteoblastic bone formation, which were decreased in wild-type mice by unloading, were not altered in OPN(-/-) mice. These observations indicate that the presence of OPN is a prerequisite for the activation of osteoclastic bone resorption and for the reduction in osteoblastic bone formation in unloaded mice. Thus, OPN is a molecule required for the bone loss induced by mechanical stress that regulates the functions of osteoblasts and osteoclasts.

Amino Acids↗
PubMed2001

Parathyroid hormone-induced bone resorption does not occur in the absence of osteopontin.

Osteopontin is an RGDS-containing protein that acts as a ligand for the alpha(v)beta(3) integrin, which is abundantly expressed in osteoclasts, cells responsible for bone resorption in osteopenic diseases such as osteoporosis and hyperparathyroidism. However, the role of osteopontin in the process of bone resorption has not yet been fully understood. Therefore, we investigated the direct function of osteopontin in bone resorption using an organ culture system. The amount of (45)Ca released from the osteopontin-deficient bones was not significantly different from the basal release from wild type bones. However, in contrast to the parathyroid hormone (PTH) enhancement of the (45)Ca release from wild type bones, PTH had no effect on (45)Ca release from organ cultures of osteopontin-deficient bones. Because PTH is located upstream of receptor activator of NF-kappaB ligand (RANKL), that directly promotes bone resorption, we also examined the effect of RANKL. Soluble RANKL with macrophage-colony stimulating factor enhanced (45)Ca release from the bones of wild type fetal mice but not from the bones of osteopontin-deficient mice. To obtain insight into the cellular mechanism underlying the phenomena observed in osteopontin-deficient bone, we investigated the number of tartrate-resistant acid phosphatase (TRAP)-positive cells in the bones subjected to PTH treatment in cultures. The number of TRAP-positive cells was increased significantly by PTH in wild type bone; however, no such PTH-induced increase in TRAP-positive cells was observed in osteopontin-deficient bones. These results indicate that the absence of osteopontin suppressed PTH-induced increase in bone resorption via preventing the increase in the number of osteoclasts in the local milieu of bone.

Animals↗
PubMed2001

CD34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages.

The CD34 antigen serves as an important marker for primitive hematopoietic cells in therapeutic transplantation of hematopoietic stem cells (HSC) and gene therapy, but it has remained an open question as to whether or not most HSC express CD34. Using a competitive long-term reconstitution assay, the results of this study confirm developmental changes in CD34 expression on murine HSC. In fetuses and neonates, CD34 was expressed on Lin(-)c-Kit(+) long-term repopulating HSC of bone marrow (BM), liver, and spleen. However, CD34 expression on HSC decreased with aging, and in mice older than 10 weeks, HSC were most enriched in the Lin(-)c-Kit(+)CD34(-) marrow cell fraction. A second transplantation was performed from primary recipients who were transplanted with neonatal Lin(-)c-Kit(+) CD34(high) HSC marrow. Although donor-type HSC resided in CD34-expressing cell fraction in BM cells of the first recipients 4 weeks after the first transplantation, the stem cell activity had shifted to Lin(-)c-Kit(+)CD34(-) cells after 16 weeks, indicating that adult Lin(-)c-Kit(+)CD34(-) HSC are the progeny of neonatal CD34-expresssing HSC. Assays for colony-forming cells showed that hematopoietic progenitor cells, unlike HSC, continue to express CD34 throughout murine development. The present findings are important because the clinical application of HSC can be extended, in particular as related to CD34-enriched HSC and umbilical cord blood HSC.

Age Factors↗
PubMed2001

Younger patients have a higher rate of recovery of taste function after middle ear surgery.

BACKGROUND: Although the chorda tympani nerve (CTN) is frequently damaged during tympanoplasty, little attention has been given to the patients' symptoms and taste function. OBJECTIVE: To investigate patients' symptoms and the functional recovery of taste after surgery using electrogustometry (EGM). DESIGN: Prospective study. PATIENTS: Analysis of 163 ears of 156 patients who underwent middle ear surgery from April 1997 through December 1999. There were 18 ears with noninflammatory diseases, 80 with chronic otitis media, and 65 with cholesteatoma. The patients' taste functions were examined 2 days before surgery and 2 weeks and 6 months after surgery. MAIN OUTCOME MEASURES: The taste disturbance before and after middle ear surgery and the relationship between age and the recovery rate of CTN function. RESULTS: Two weeks after surgery, the mean EGM threshold was elevated in all groups regardless of preservation or section of the CTN. Numbness in the tongue and taste disturbance were more frequently found in patients with preservation of CTN than in those with section of the CTN (P =.008 and P =.001, respectively). In patients with preservation of the CTN, 6 months after surgery, the recovery rate of EGM threshold was 83% in those aged 20 years or younger (P =.008 compared with the 2 older groups), 53% in the those aged 21 to 40 years, and 44% in those aged 41 to 60 years. CONCLUSION: Age is an important factor for recovery of taste function after middle ear surgery, which is useful information when explaining complications to patients.

Adolescent↗
PubMed2001

Transcriptional suppression of Sox9 expression in chondrocytes by retinoic acid.

SOX9 is a transcription factor that is expressed in chondrocytes and regulates expression of chondrocyte phenotype related genes. Expression of these genes is known to be suppressed by retinoic acid (RA). We, therefore, examined whether the Sox9 gene expression is regulated by RA in chondrocytes. RA treatment suppressed Sox9 mRNA expression in primary chondrocytes prepared from newborn mouse rib cartilage within 12 h and this suppression lasted at least up to 24 h. The RA suppression of Sox9 mRNA levels was dose-dependent starting at 0.5 microM with a maximum at 1 microM. Nuclear run-on assays revealed that RA reduced the rate of transcription of Sox9 gene. Finally, Western blot analysis indicated that RA suppressed SOX9 protein levels in these chondrocytes. Furthermore, overexpression of SOX9 reversed RA suppression of Col2a1 enhancer activity. These observations indicate that RA suppresses Sox9 gene expression in chondrocytes at least in part through transcriptional events. J. Cell. Biochem. Suppl. 36: 71-78, 2001.

Animals↗
PubMed2001

Characterization and subcellular localization of KCNQ1 with a heterozygous mutation in the C terminus.

Numerous mutations in KCNQ1, a gene encoding the alpha -subunit of cardiac delayed rectifier potassium channels, have been found in long QT syndrome (LQTS). Among them, several mutations in the C terminus have been shown to cause autosomal recessive or subclinical autosomal dominant LQTS. Here, we report a heterozygous mutation, T587M, which is also in the KCNQ1 C-terminal domain. The same mutation was found in three independent probands that were clearly symptomatic with family history of cardiac sudden death. Functional assay using a heterologous expression system with a mammalian cell line (COS7 cells) revealed that the mutant displayed neither functional channels when expressed alone nor dominant-negative effect when co-expressed with wild-type (WT) KCNQ1. To examine the cellular trafficking of KCNQ1, green fluorescent protein (GFP) was tagged to the cytoplasmic C terminus of WT or mutant KCNQ1. This procedure did not affect the essential properties of expressed WT KCNQ1 channels. On confocal microscopic images, GFP-tagged WT KCNQ1 showed a plasma membrane fluorescence pattern, whereas the GFP-tagged mutant showed a perinuclear fluorescence pattern. Co-expression of the mutant with GFP-tagged WT KCNQ1 did not influence its normal cellular transport. Therefore, the T587M mutant cannot traffic to the plasma membrane and may form no subunit assembly with WT KCNQ1. These findings provide a novel molecular basis for the clinical finding that this C-terminal mutation produced a severe form of RWS-type LQTS.

Adolescent↗
PubMed2001

Laparoscopic splenectomy for a lymphangioma of the spleen: report of a case.

We present the first case report of a successful laparoscopic complete excision of a splenic lymphangioma. The splenic tumor was preoperatively diagnosed to be a lymphangioma by the combined modalities of ultrasonography, computed tomography, magnetic resonance imaging, and angiography. A laparoscopic splenectomy was subsequently performed and the pathological examination of the mass confirmed the diagnosis of a lymphangioma. Based on the above findings, a laparoscopic splenectomy is recommended when a splenic tumor is suspected to be either benign or borderline.

Aged↗
PubMed2001

Transient suppression of core-binding factor alpha 1 expression by basic fibroblast growth factor in rat osteoblast-like osteosarcoma ROS17/2.8 cells.

Core-binding factor alpha 1 (Cbfa1) is considered a prerequisite transcription factor for osteoblastic differentiation, based on experiments performed in knock-out mice. Cbfa1 binding sequences have been located in the promoter regions of various osteoblast phenotype-related genes, which are also known to be regulated by fibroblast growth factor (FGF). To elucidate the molecular mechanisms of FGF regulation of osteoblasts, we investigated the effects of FGF on Cbfa1 expression in osteoblast-like cells. We found that basic FGF, at 10 ng/ml, suppressed Cbfa1 mRNA expression in ROS17/2.8 cells. To detect the two distinct mRNA species that give rise to Pebp2alphaA/Cbfa1 and Osf2/Til-1/Cbfa1, we used semi-quantitative reverse transcription-polymerase chain reaction analysis using specific sets of primers. This analysis indicated that treatment with FGF transiently decreased the abundance of only the Osf2/Til-1/Cbfa1 isoform and not the Pebp2alphaA/Cbfa1 isoform in ROS17/2.8 cells. The suppression of Osf2/Til-1/Cbfa1 was time dependent and was observed up until 48h, and Cbfa1 mRNA levels then returned closer to control levels. Dose-dependent suppression by FGF was observed only with regard to mRNA levels of Osf2/Til-1/Cbfa1 and not for Pebp2alphaA/Cbfa1 mRNA. These data indicate that FGF suppresses Cbfa1 mRNA expression and that the Osf2/Til-1/Cbfa1 isoform is the specific target of FGF regulation in osteoblastic osteosarcoma ROS17/2.8 cells.

Animals↗
PubMed2001

Analysis of microcystins in sediments using MMPB method.

During the course of study on the detoxification of microcystins, the adsorption on sediments in the natural environment was investigated. Because it was very difficult to extract microcystins from sediments using conventional techniques, a physicochemical screening method, the MMPB (2-methyl-3-methoxy-4-phenylbutyric acid) method, including ozonolysis and mass spectrometric detection was developed. This method consisted of the following operations: lyophilized sediments were suspended in methanol and MMPB-d(3) as an internal standard was added to this suspension, which was cooled at -78 degrees C with vigorous stirring and then treated with a stream of ozone/oxygen. After centrifugation, an aliquot of the reaction solution was subjected to EI (electron ionization)-GC/MS analysis after methylation with 14% BF(3)-methanol and liquid-liquid extraction. The established method had a potential for the analysis of microcystins in sediments that are difficult to analyze using conventional methods. Finally, this method was applied to sediment samples collected in Japanese lakes and six of the eleven samples showed positive results. The obtained results clearly indicated that the adsorption on sediments contributes to the detoxification of microcystins under natural conditions.

Adsorption↗
PubMed2001

Development of a new enzyme-linked immunosorbent assay for the detection of anti-M2 in primary biliary cirrhosis.

We developed a new enzyme-linked immunosorbent assay (ELISA) for the detection of antimitochondrial antibody (AMA)-M2 in sera from patients with primary biliary cirrhosis (PBC), using 2-oxo-acid dehydrogenase complex (2-OADC) purified from porcine myocardium as the antigen source. The immunoreactivity was tested in a total of 354 sera, including 63 sera from patients with PBC by our ELISA. In the sera, indirect immunofluorescence for AMA, former ELISA for anti-pyruvate dehydrogenase complex (PDC) and immunoblot assay were performed, respectively. Of the 63 sera from patients with PBC, 51 sera (81.0%) were positive for anti-M2 in the new ELISA. Thirty-eight of the 63 sera (60.3%) were positive for anti-PDC in the former ELISA; the difference was significant between them (P=0.011). None of the 291 control sera from healthy volunteers showed reactivity against 2-OADC in the new ELISA. Moreover, in comparison with the results of immunoblot analysis, sensitivity and specificity in our ELISA to the sera from patients with PBC were 100 and 92.3%, respectively. Our results indicate that the new ELISA for anti-M2 using 2-OADC is simple, rapid and sensitive enough for the detection of AMA specific to PBC.

Journal Article↗
PubMed2001

Disopyramide and its metabolite enhance insulin release from clonal pancreatic beta-cells by blocking K(ATP) channels.

In an insulin-secreting pancreatic beta-cell line (MIN6), insulin release was caused by disopyramide, an antiarrhythmic drug with Na-channel blocking action, and its main metabolite mono-isopropyl disopyramide (MIP). Insulin secretion, measured as immunoreactive insulin (IRI), was accelerated to 265.7% of the control by disopyramide and to 184.4% by MIP, with half-effective concentrations (EC50) of 30.9 +/- 1.5 microM and 92.4 +/- 2.2 microM. We tested the possibility that these drugs induce insulin release by inhibiting ATP-sensitive K+ (K(ATP)) channels of MIN6 cells. In the cell-attached or ATP-free inside-out mode with patch membranes on MIN6 cells, K-selective channels were recorded with unitary conductance of 70.5 +/- 3.5 pS (150 mM external K+ ions at room temperature). The channels were concluded to be MIN6-K(ATP) channels because they were closed by extracellular high glucose (11.0 mM) or glibenclamide (200 nM) and were reversibly activated by diazoxide (50 microM). In the inside-out patch mode, they were inhibited by micromolar ATP. In both cell-attached and insideout mode, disopyramide and MIP inhibited single MIN6-K(ATP) channels. In the inside-out mode, they produced a dose-dependent inhibition of channel activity: the half-blocking concentrations (IC50) were 4.8 +/- 0.2 microM for disopyramide and 40.4 +/- 3.1 microM for MIP. It was therefore concluded that both agents exert insulinotrphic effect through the inhibition of membrane K(ATP) channels in MIN6 cells.

ATP-Binding Cassette Transporters↗
PubMed2001

Prompt and durable hematopoietic reconstitution by unrelated cord blood transplantation in a child with Fanconi anemia.

We describe here the case of an 8-year-old girl with Fanconi anemia (FA) whose hematopoiesis was successfully restored by unrelated umbilical cord blood (UCB) transplantation. The patient became resistant to androgen therapy, and developed intracranial hemorrhage and dyserythropoiesis. Her hematopoietic recovery after the transplantation was excellent and a complete chimerism has been durably maintained. UCB should be considered as a stem cell source for transplantation when a patient with FA does not have an HLA-identical unaffected sibling donor.

Child, Preschool↗
PubMed2001

ik3-1/Cables is a substrate for cyclin-dependent kinase 3 (cdk 3).

p70ik3-1 (a 70-kDa protein) contains a cyclin box, and binds to p35cdk3 in vivo and in vitro [Matsuoka, M., Matsuura, Y., Semba, K. & Nishimoto, I. (2000) Biochem. Biophys. Res. Commun. 273, 442-447]. In spite of its structural similarity to cyclins, p70ik3-1 does not activate cyclin-dependent kinase 3 (cdk3)-mediated phosphorylation of pRb, histone H1, or the C-terminal domain of RNA polymerase II. Here, we report that Ser274 of p70ik3-1 is phosphorylated by cdk2 or cdk3 bound to cyclin A and to cyclin E in vitro. We also found that in COS7 cells in which cyclin E and cdk3 were ectopically overexpressed, the phosphorylation level of Ser274 in coexpressed p70ik3-1 is upregulated. We therefore conclude that p70ik3-1 is a substrate for cdk3-mediated phosphorylation.

Animals↗
PubMed2001

Inhibitory effect of interleukin 3 on early development of human B-lymphopoiesis.

Interleukin 3 (IL-3) is known as a stimulator of proliferation and differentiation of non-lymphoid cells, but information about the activity of IL-3 in lymphopoiesis is limited. In the present study, we examined the effect of IL-3 on human B-lymphopoiesis using the co-culture system on a murine stromal cell line, MS-5, with stem cell factor (SCF) and granulocyte colony-stimulating factor (G-CSF). Although a large number of CD45+CD19+ B cells were generated in the co-culture of cord blood CD34+ cells, the addition of IL-3 to the co-culture suppressed the B-cell generation in a dose-dependent manner. In the co-culture, CD34+IL-3 receptor alpha-chain (IL-3Ralpha)+ cells, but not IL-3Ralpha- cells, produced B cells, and IL-3 suppressed the B-cell generation from CD34+IL-3Ralpha+ cells, suggesting that IL-3 exerts an inhibitory effect through the binding to IL-3Ralpha on CD34+ cells. Because the delayed addition of IL-3 and the short-term exposure to IL-3 showed that IL-3 acted as an inhibitor at an early stage of B-cell development from CD34+ cells, before the generation of CD19+ B cells, the effect of IL-3 on the early development of B cells from immature CD34+CD38- cells was examined. Individual colonies, produced from clone-sorted CD34+CD38- cells by SCF, thrombopoietin and a complex of IL-6/soluble IL-6R with or without IL-3, were divided into two fractions and analysed for B-cell potential using co-culture on MS-5 with SCF and G-CSF, and haematopoietic potential using methylcellulose clonal culture. Although some colonies cultured without IL-3 showed both B-cell and haematopoietic potential, all the colonies cultured with IL-3 showed only haematopoietic potential. These results indicate that IL-3 has an inhibitory effect on the B-cell generation from human lymphohaematopoietic progenitor cells.

Animals↗
PubMed2001

Risk of alanine aminotransferase flare-up among asymptomatic hepatitis C virus RNA carriers: a 10-year follow-up study.

AIMS: The aim of this study was to determine the cumulative rate of flare-up of serum alanine aminotransferase (ALT) level during a 10-year follow-up period, and characterize the clinical, virologic features in 120 hepatitis C virus (HCV) RNA-positive asymptomatic carriers with persistently normal ALT levels for 6 months. RESULTS: All flare-up cases occurred during the first 5 years of the present study, 27.4% of carriers showed ALT flare-up during this period, but none in the second half of the study. Multivariate analysis showed that C100-3 antibody (Ab) and anti-human T cell leukemia virus (HTLV)-I Ab were two independent and significant predictors of ALT flare-up in hepatitis C virus (HCV) RNA asymptomatic carriers (P = 0.04, P = 0.03, respectively). Liver biopsy was performed in 44 patients (11 with flare-up of ALT level, whereas 33 had normal ALT levels). Histological features of chronic hepatitis with lymphoid infiltration in the portal tracts were commonly observed in all specimens, and no differences were noted between the flare-up ALT group and the persistently normal ALT group. CONCLUSIONS: Our results indicated that flare-up of ALT levels in asymptomatic HCV-RNA carriers with normal ALT levels occurs during the first 5 years of diagnosis, and that the presence of C100-3 and anti-HTLV-I antibodies are good predictors of a transient rise in ALT.

Adult↗
PubMed2001

Panic disorder cases in Japanese-Brazilians in Japan: their ethnic and cultural confusion.

The comparatively high salaries made in Japan are attractive to many Japanese-Brazilians. The number of individuals from this ethnic group being treated in Japanese mental hospitals has increased. We hypothesized that Japanese-Brazilian patients with panic disorders adjusted better to Japanese society and culture than those with other mental disorders. The subjects in the present study are 40 Japanese-Brazilian patients undergoing treatment at the Department of Psychiatry at Jichi Medical School, Japan, from May 1990 to September 1998. Patients were divided into a panic disorder group, a schizophrenic group, a mood disorder group and a neurosis group. Demographic data (Japanese language ability, duration of residence in Japan etc.) were collected. A comparison was made among the four groups. Patients in the panic disorder group showed a significant tendency to be fluent speakers of Japanese. Patients in the panic disorder group also had been in Japan for a significantly longer period of time than those in the other three groups. Japanese ability and length of residence in Japan rule out exacerbating factors due to a foreign living environment. Panic disorder patients usually have resolved the problems inherent in living and working in a foreign country. In general, Japanese-Brazilians are more comfortable both financially and socially in Japan than other foreign laborers because of their cultural and family background. The emotional conflict experienced by such patients may result from concern over whether to live in Brazil or Japan in the future. Their ethnic and cultural identity may be confused, fluctuating between identifying with Brazil and with Japan, and this may cause vague feelings of anxiety.

Adult↗
PubMed2001

Hemodialysis paradox: survey on the incidence rate of hepatocellular carcinoma in antihepatitis virus C-antibody-positive chronic hemodialysis patients.

The prevalence of antihepatitis virus C (HCV) antibodies in chronic hemodialysis (HD) patients is higher than in normal populations, and yet hepatocellular carcinoma (HCC) is infrequent in chronic HD patients who are HCV antibody positive. In this study, we investigated the characteristics of HCV-antibody-positive patients with HCC on chronic HD. A total of 6,366 cases of HCV-antibody-positive patients on chronic HD therapy was analyzed on the basis of answers to questionnaires on the incidence rate of HCC in 314 Japanese dialysis institutions. HCC was a complication in 114 of 6,222 (1.8%) HCV-antibody-positive patients, and cirrhosis was a complication in 536 of 6,242 (8.6%). The incidence rate of both complications was significantly higher in males than in females, and the incidence rate in the chronic HD patients was much lower than in normal populations. Specific immunological status in patients on chronic HD therapy may be an important key for preventing the progression of chronic HCV hepatitis. However, further research is needed because this study was preliminary and excluded the type of HCV virus, pathological findings, and laboratory data.

Carcinoma, Hepatocellular↗