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Biomedical subjects

T Tsukamoto

Publications and source records attributed to T Tsukamoto.

At least 127 records · Page 7Linked to original sources
PubMed2000

Contributing factors influencing the functional outcome of floating knee injuries.

The purpose of the present study was to retrospectively review the floating knee injuries treated at our institute and to determine various factors, such as severity of soft-tissue or skeletal injuries, site of fractures, and treatment methods that may significantly influence the final functional result in these injuries. Between 1986 and 1996, 65 patients with 66 floating knee injuries were treated in our institution. Among 66 fractures of the femur, 19 (29%) were open. There were 43 open tibial fractures. Fifty cases were Fraser type I floating knee fractures, 7 were type IIa, 2 were type IIb, and 7 were type IIc. In 63 cases (95%), both bones had been surgically stabilized with interlocked nails, Ender pins, plates, screws with/without pinning, or external fixations. Final functional results were evaluated according to Karlström and Olerud's criteria. Satisfactory results were rated as cases with excellent or good results. The mean follow-up time was 16.6 months range, (12-50 months). We assessed various factors influencing functional results, including Fraser type, severity of open injury grade (Gustilo) in both fractures, combination of open/closed injuries, fracture types (AO/ASIF type), existence of multiple trauma, neurovascular injuries, polyskeletal trauma, and stabilizing method or operation timing of both fractures. Satisfactory rates in Fraser type I and type II were 64% and 25%, respectively (P= .02). The satisfactory rate in closed, grade I+II, and grade III injuries of the femoral fractures was 53.2%, 81.8%, and 25%, respectively (grade I+II vs. grade III: P < .03). There were no significant correlations between the functional result and the following factors: soft-tissue injuries of the tibia; the fracture pattern of both fractures; the combination of open/closed injuries in each fracture; injury severity score; the existence of neurovascular injuries and double femoral fractures; treatment methods; and operation timing. Severity of damage to the knee joint and open injuries in the thigh were found to be significant factors contributing to the functional outcome in floating knee injuries.

Adolescent↗
PubMed2000

[Occurrence of angioimmunoblastic T-cell lymphoma six months after onset of Lyme disease].

A 68-year-old man, who had suffered a tick bite one week previously, consulted his home doctor because of fever and an erythematous rash around the bite scar. He underwent a skin biopsy, and Borrelia garinii was detected, from which Lyme disease was diagnosed. He received amoxicillin for two weeks and his symptoms disappeared. After 6 months he noticed swelling of his cervical, axillary and inguinal lymph nodes. A biopsy sample was taken from a left cervical lymph node, and this revealed angioimmunoblastic T-cell lymphoma. The patient achieved a complete remission after chemotherapy. The relationship between Lyme disease and lymphoma is discussed.

Aged↗
PubMed2000

Portal vein stenting to treat portal vein tumor thrombus in hepatocellular carcinoma.

Tumor thrombus in the portal vein is refractory to therapy and a serious negative prognostic factor in hepatocellular carcinoma. We describe herein the case of a patient in whom an expandable metallic stent was successfully employed to treat portal vein tumor thrombus by restoring blood flow. An expandable metallic stent was implanted in a patient with severe main portal vein stenosis caused by tumor thrombus protruding from the left portal vein branch. Immediately after stent placement, the stenotic lesion was effectively dilated, the portal blood flow restored, and portal hypertension relieved. Angiography, computed tomography, and doppler ultrasonography done 2 months after the stent placement revealed continued patency of the portal vein. Thus, intraportal placement of an expandable metallic stent appears to be an effective treatment for major portal vein tumor

Aged↗
PubMed1999

Formation of peroxisomes from peroxisomal ghosts in a peroxisome-deficient mammalian cell mutant upon complementation by protein microinjection.

Most mammalian cell strains genetically deficient in peroxisome biogenesis have abnormal membrane structures called ghosts, containing integral peroxisomal membrane protein, PMP70, but lacking the peroxisomal matrix proteins. Upon genetic complementation, these mutants regain the ability of peroxisome biogenesis. It is postulated that, in this process, the ghosts act as the precursors of peroxisomes, but there has been no evidence to support this. In the present study, we investigated this issue by protein microinjection to a mutant Chinese hamster ovary cell line defective of PEX5, encoding a peroxisome-targeting signal receptor. When recombinant Pex5p and green fluorescent protein (GFP) carrying a peroxisome-targeting signal were co-injected into the mutant cells, the GFP fluorescence gathered over time to particulate structures where PMP70 was co-localized. This process was dependent on both Pex5p and the targeting signal, and, most importantly, occurred even in the presence of cycloheximide, a protein synthesis inhibitor. These findings suggest that the ghosts act as acceptors of matrix proteins in the peroxisome recovery process at least in the PEX5 mutant, and support the view that peroxisomes can grow by incorporating newly synthesized matrix proteins.

ATP-Binding Cassette Transporters↗
PubMed1999

Involvement of a cellular glycolytic enzyme, phosphoglycerate kinase, in Sendai virus transcription.

In vitro mRNA synthesis of Sendai virus is almost entirely dependent on the addition of cellular proteins (host factors). Previous studies indicated that the host factor activity from bovine brain was resolved into at least two complementary fractions, one of which may be tubulin. In this study, the host factor activity that stimulates the transcription in the presence of tubulin was further purified from bovine brain. This fraction was found to contain at least two complementary factors, and one of them was purified to a single polypeptide chain with an apparent M(r) of 46,000 (p46). From the amino acid sequence, biochemical, and immunological analyses, p46 was identified as a glycolytic enzyme, phosphoglycerate kinase (PGK). Purified native PGK from rabbit and yeast, and a recombinant human PGK substituted for p46. Although, as previously suggested, tubulin was involved in the transcription initiation complex formation by being integrated into the complex, p46 and its complementary factor had little effect on the complex formation. On the other hand, when p46 and the complementary factor were added to the RNA chain elongation reaction from the isolated initiation complex formed with tubulin, mRNA synthesis was dramatically stimulated. The enzymatic activity per se of PGK did not seem to be required for its activity. West-Western blot analysis showed that PGK could directly interact with tubulin. These data suggest that PGK stimulates Sendai virus mRNA synthesis at the elongation step, probably through its interaction with tubulin in the initiation complex.

Amino Acid Sequence↗
PubMed1999

Chiral resolution of basic drugs by capillary electrophoresis with new glycosaminoglycans.

New glycosaminoglycans, fucose-containing glycosaminoglycan (FGAG) and depolymerized holothurian glycosaminoglycan (DHG), were investigated as chiral additives for the separation of drug enantiomers by capillary electrophoresis. The average molecular masses of FGAG and DHG were estimated to be about 59,000 and 14,000, respectively. A variety of basic drug enantiomers were resolved using 10 mM phosphate buffer, pH 5.0, containing 3% FGAG or DHG. Since chiral recognition properties of FGAG and DHG are different, some drug enantiomers were only separated by using FGAG or DHG. With regard to comparison of chiral recognition abilities of FGAG and DHG with other chiral selectors, tolperisone and eperisone enantiomers were not separated with alpha- or beta-cyclodextrin, or heparin as the chiral additives, but were separated with FGAG and DHG. The results obtained reveal that FGAG and DHG are useful as the chiral selectors for separations of drug enantiomers by CE, and that they could be complementarily used with other chiral additives.

Buffers↗
PubMed1999

Clinical significance of prior hepatitis B virus infection in patients with hepatitis C virus-related hepatocellular carcinoma.

BACKGROUND: The effect of prior hepatitis B virus (HBV) infection on the clinicopathologic findings for patients with hepatitis C virus (HCV) RNA and hepatocellular carcinoma (HCC) is still unclear. METHODS: Of 59 patients who underwent liver resection for HCV-related HCC (</=2.0 cm in greatest dimension) without metastases between 1990 and 1997, 38 patients had anti-HB core antibody (anti-HBc) and did not have HB surface antigen in their sera (Group 1). Their clinicopathologic findings and outcomes after surgery were compared with those of the remaining 22 patients without anti-HBc (Group 2). RESULTS: The proportion of well-differentiated HCC was significantly lower in Group 1 than in Group 2 (P = 0.0214). The percentage of patients with cirrhosis was significantly lower in Group 1 than in Group 2 (P = 0.0228). The cumulative survival rate was significantly lower in Group 1 than in Group 2 (P = 0.0224). The risk ratio of anti-HBc for shorter survival time was 3.817. CONCLUSIONS: HCC more often developed before cirrhosis in patients with HCV RNA and anti-HBc than in patients positive for HCV RNA alone. Prior HBV infection was a risk factor for poor outcome after liver resection for patients infected with HCV.

Aged↗
PubMed1999

Functional heterogeneity of C-terminal peroxisome targeting signal 1 in PEX5-defective patients.

To investigate mechanisms related to functions of the peroxisome targeting signal (PTS) 1 receptor, Pex5p, we analyzed peroxisome matrix protein import in fibroblasts from three patients with peroxisome biogenesis disorders, all with different mutations in the PEX5 gene. The patients 2-01 (Zellweger syndrome) and 2-05 (neonatal adrenoleukodystrophy) have the reported mutations, R390X and N489K, and patient 2-03 (infantile Refsum disease) has a newly identified mutation, S563W. Fibroblasts from 2-03 (S563W) were detected in both PTS1 and PTS2 imports despite the PEX5 defect, findings in contrast with fibroblasts from 2-05 (N489K) severely defective in PTS1 import and those from 2-01 (R390X) severely defective in both PTS1 and PTS2. The PTS1 receptor in 2-03 is functional for only the C-terminal -SKL sequence (acyl-CoA oxidase) and had little or no function for C-terminal -AKL (D-bifunctional protein and sterol carrier protein 2) and -KANL (catalase) sequences, respectively. After transfection of these mutated PEX5 cDNA into the PEX5-defective CHO mutant, transformants of ZP102 revealed that each mutation was responsible for each dysfunction of the PTS1 import. It seems apparent that -AKL and -KANL are poorer variants of PTS1 and are likely to be more susceptible to effects of mutation of its receptor, Pex5p.

17-Hydroxysteroid Dehydrogenases↗
PubMed1999

Cell-cell dissociation upon epithelial cell scattering requires a step mediated by the proteasome.

During development, tissue repair, and tumor metastasis, both cell-cell dissociation and cell migration occur and appear to be intimately linked, such as during epithelial "scattering." Here we show that cell-cell dissociation during scattering induced by hepatocyte growth factor (HGF) or activation of the temperature-sensitive v-Src tyrosine kinase in MDCK cells can be blocked by inhibiting the proteasome with lactacystin and MG132. Although both proteins of the tight junction and the adherens junction redistributed during cell scattering, proteasome inhibitors largely prevented this process, resulting in the stabilization of Triton X-100-insoluble tight junction proteins as well as adherens junction proteins at sites of cell-cell contact. Proteasome inhibition also led to a decrease of E-cadherin turnover in (35)S-labeled cells. In addition, proteasome inhibition partly preserved cell polarity, as determined by the subcellular distribution of Na(+),K(+)-ATPase (basolateral marker) and gp135 (apical marker), and the structure of the subcortical actin ring, both of which are normally disrupted during scattering. However, cells were able to establish focal contacts, and single cell migration toward HGF was unaffected by proteasome inhibition in quantitative assays, indicating that cell-cell dissociation during scattering occurs independently of anchorage-dependent cell migration. Thus, a proteasome-dependent step during scattering induced by HGF and pp60(v-Src) appears to be essential for cell-cell dissociation, disassembly of junctional components, and (at least indirectly) it also plays a role in the loss of protein polarity.

Acetylcysteine↗
PubMed1999

Chemosensitivity of micrometastases and circulating tumor cells to uracil and tegafur as evaluated using LacZ gene-tagged Lewis lung carcinoma cell.

The chemosensitivity of the sequence of steps responsible for metastasis formation including circulating tumor cells and micrometastases to a 5-fluorouracil derivative (UFT) was examined with a novel micrometastasis model featuring Lewis lung carcinoma cells tagged with the bacterial LacZ gene and hygromycinR gene (hygR). Metastases in the lung could be specifically detected at the single-cell level by X-Gal staining after inoculation of LacZ-transfected tumor cells. Spontaneous metastasis in mice bearing subcutaneous primary tumors was significantly inhibited by UFT at doses of 15-20 mg/kg when it was orally administered from day 14, during the early stage of micrometastasis formation, but not by late administration from day 22. Experimental pulmonary metastasis was also inhibited without significant toxic side effects by oral administration of UFT at a daily dose of 20 mg/kg from day 4, when the tumor cells start new growth in the lung, but not by daily treatment from 8 days after intravenous injection. Oral administration of UFT had no effect on tumor cell arrest in the lung as detected by X-Gal staining. Furthermore, PCR analysis revealed that circulating tumor cells in the peripheral blood of mice bearing primary tumors after subcutaneous injection of hygR-tagged tumor cells were significantly reduced by the oral administration of UFT for 7 days in a dose-dependent manner. These results indicate that circulating tumor cells in the peripheral blood and micrometastases in the initial growth phase in the lungs are sensitive to this chemotherapeutic agent, and suggest that micrometastasis formation can be effectively inhibited by long-term oral administration of anticancer agents with minimal toxic side effects.

Administration, Oral↗
PubMed1999

Effects of low dose catechol on glandular stomach carcinogenesis in BALB/c mice initiated with N-methyl-N-nitrosourea.

The effects of low dose catechol administration in the diet on stomach carcinogenesis in mice after initiation with N-methyl-N-nitrosourea (MNU) in the drinking water were investigated. Male, 6-week-old, BALB/c mice were given MNU in the drinking water intermittently for 1-week periods at 1-week intervals for a total of 3 weeks at a concentration of 120 ppm (groups 1 and 3). Groups 2 and 4 served as non-initiated controls. From week 7, groups 1 and 3 were divided into four subgroups and the mice were fed on a diet containing 4 ppm (groups la and 3a), 20 ppm (groups 1b and 3b), 100 ppm (groups 1c and 3c), 500 ppm (groups 1d and 3d) or 0 ppm (groups 2 and 4) catechol for 44 weeks. At week 50, appreciably enhanced development of pepsinogen 1 altered pyloric glands (PAPG) was noted in groups 1c and 1d. The incidences of adenomatous hyperplasia and carcinomas were not affected in any of the catechol-treated groups as compared with corresponding controls on a basal diet. Thus, the administration of catechol in the diet at low doses enhanced only preneoplastic lesion development and not neoplastic lesion development. From these results, we conclude that the biological significance of the catechol promoting effect at probable human exposure levels on gastric cancer is probably limited, while the PAPG may be a sensitive endpoint lesion for mouse glandular stomach carcinogenesis.

Adenocarcinoma↗
PubMed1999

Identification of a novel gene, OASIS, which encodes for a putative CREB/ATF family transcription factor in the long-term cultured astrocytes and gliotic tissue.

Gliosis is a characteristic response of astrocytes to inflammation and trauma of the central nervous system (CNS). To study the mechanisms underlying gliosis, we performed differential display screening for genes specifically induced in long-term cultured astrocytes used as an in vitro gliosis model. We identified and characterized a gene (named OASIS, for old astrocyte specifically-induced substance) expressed in long-term cultured mouse astrocytes, or 'old astrocytes (OA)'. The OASIS gene encoded a putative transcription factor belonging to the cyclic AMP responsive element binding protein/activating transcription factor (CREB/ATF) gene family, with homology to box B-binding factor-2 (BBF-2), a Drosophila transcription factor. Its expression was developmentally regulated; OASIS mRNA was primarily expressed in the salivary gland and cartilage in the mouse embryo and it was transiently upregulated in the brain during postnatal two weeks. The expression became weaker in the adult brain. We also demonstrated that an expression of the OASIS mRNA was induced in response to the cryo-injury of the mouse cerebral cortex. The distribution pattern of the OASIS-positive cells in the injured cortex was very similar to that of the glial fibrillary acidic protein (GFAP)-positive cells. These results suggest that OASIS protein may play a role in gliotic events.

Activating Transcription Factors↗
PubMed1999

The significance of microsatellite instability in predicting the development of metachronous multiple colorectal carcinomas in patients with nonfamilial colorectal carcinoma.

BACKGROUND: Patients with metachronous multiple colorectal carcinomas have been reported to have a higher frequency of a family history of colorectal carcinoma, associated colorectal adenomas, and extracolonic malignancies. These clinicopathologic factors also are considered to be related to the development of metachronous multiple colorectal carcinomas after surgery for colorectal carcinoma. In this article, the authors investigated whether genetic markers such as microsatellite instability (MSI) were helpful in predicting the development of metachronous multiple colorectal carcinomas. METHODS: Between 1990-1997, 312 colorectal carcinoma patients underwent yearly surveillance colonoscopy after surgery. Among these patients, there were 19 with nonfamilial colorectal carcinoma in whom metachronous multiple colorectal carcinomas were diagnosed during the yearly surveillance colonoscopy. A control group was comprised of 28 patients who did not demonstrate either synchronous or metachronous carcinomas over a period of > or =5 years. Six microsatellite markers (D2S123, D3S1029, D3S1611, TP53, Mfd26, and Mfd36) were used to determine MSI by polymerase chain reaction. RESULTS: The frequency of MSI positive cases was significantly higher in patients with sporadic metachronous multiple colorectal carcinomas than in those with a single carcinoma (17/19 [89%] vs. 4/28 [14%]; P<0.0001). In tumors occurring in the distal colon and rectum, the percentage of MSI positive carcinomas was significantly higher in the patients with metachronous multiple carcinomas than in those with a single carcinoma (13/15 [87%] vs. 0/19 [0%]; P<0.0001). No such difference was observed in the proximal colon. CONCLUSIONS: Based on the findings of the current study, the analysis of MSI in sporadic carcinomas of the distal colon and rectum may be helpful in predicting the development of metachronous multiple colorectal carcinomas.

Colonoscopy↗
PubMed1999

Identification of receptor genes in renal cell carcinoma associated with angiogenesis by differential hybridization technique.

To screen the receptor genes in renal cell carcinoma (RCC) associated with angiogenesis, we performed differential hybridization of the cDNA library of membrane-type protein tyrosine kinases (mPTKs). Three thousand plaques of a mPTKs-enriched cDNA library were screened with mPTKs mixture probes produced from hypervascular RCC tissues and RCC cell lines. Six different cDNA fragments of the PTK genes were isolated, and the sequence analysis showed that these represented cDNAs for TIE1, KDR, FMS, FGFR-4, JAK1 and HCK. Of these genes, the expression of TIE1, KDR, and FGFR-4 was studied in RCC tissue and cell lines by Northern blot analysis. We also investigated the expression of vascular endothelial growth factor (VEGF), placenta growth factor (PlGF) and their receptor FLT-1. In all the hypervascular RCC tissues, the amounts of mRNAs for KDR and FLT-1 were increased compared to adjacent normal tissues. The TIE1 and FGFR-4 genes were also overexpressed in most of the hypervascular RCC tissues, while no mRNA of KDR, FLT-1, or TIE1 could be detected in any of the four human RCC cell lines. The amounts of the VEGF and PlGF mRNAs were increased in hypervascular RCC tissues, while VEGF mRNA was detected in the four cell lines but PlGF mRNA was not. FGFR-4 mRNA was expressed in three of the four cell lines. These results suggest that KDR, FLT-1, PlGF and TIE1 mRNAs are present in the mesenchymal cells of RCC, while VEGF and FGFR-4 genes are expressed in RCC cells themselves in vivo.

Blotting, Northern↗
PubMed1999

Detection of various epitopes of murine osteopontin by monoclonal antibodies.

We immunized rats with recombinant murine osteopontin protein and obtained four monoclonal antibodies recognizing distinct epitopes of murine osteopontin. OPN1.2 recognized the amino-terminal half of OPN, while OPN2.2, OPN2.3, and OPN3.1 recognized the carboxy-terminal half of OPN. The epitope recognized by OPN2.2 was destroyed by further cleavage of the carboxy half of OPN. The epitope recognized by OPN2.3 was located in the amino-terminal end of the carboxy half of OPN, whereas that recognized by OPN3.1 was located in the carboxy-terminal end of the carboxy half of OPN. OPN1.2 and OPN2.2 recognized thrombin-cleaved osteopontin, whereas thrombin-cleaved osteopontin was not recognized by OPN2.3 and OPN3.1. Thus, these monoclonal antibodies will be useful in structure/function studies of the role of osteopontin in murine models of disease.

Animals↗
PubMed1999

Schizophrenia and the serotonin-2A receptor promoter polymorphism.

Serotonin-2A (5-HT2A) receptors have received much investigative attention in schizophrenia because (1) several studies have shown a decrease in the number of 5-HT2A receptors in the prefrontal cortex of postmortem brains of schizophrenic patients; (2) atypical antipsychotic drugs are antagonists for 5-HT2A receptors; and (3) a positive association between a T to C polymorphism at position 102 of the 5-HT2A receptor gene and schizophrenia has been reported. A G to A polymorphism at position -1438 of the 5-HT2A receptor gene was studied in 119 schizophrenic patients and 106 healthy control subjects, all of whom were Japanese. The genotype and allele frequencies did not differ between the patients and control subjects. Furthermore, the genotype frequency did not differ according to diagnostic subtype, family history, age at onset of illness, or daily dosage of antipsychotic medication. Our results suggest that the polymorphism does not contribute to the etiology or clinical characteristics of schizophrenia. However, the gene is greater than 20 kbp in length, and thus it is possible that other areas that affect expression of the gene may vary. We found that the -1438G/A variant was in linkage disequilibrium with the T102C polymorphism.

Adult↗
PubMed1999

A novel endogenous inhibitor of phenoloxidase from Musca domestica has a cystine motif commonly found in snail and spider toxins.

Phenoloxidase inhibitor (POI), found in the hemolymph of housefly pupae, is a novel dopa-containing and cystine-rich peptide that competitively inhibits phenoloxidase with a Ki in the nanomolar range. [Tyr32]POI is a potential precursor molecule also found in the hemolymph that may be posttranslationally oxidized to the dopa-containing peptide after creation of a rigid structure. By employing both a solid-phase peptide synthesis system based on a 9-fluorenylmethoxycarbonyl strategy and a specific air oxidation technique to ensure correct folding, we have been able to synthesize [Tyr32]POI. The synthetic [Tyr32]POI was confirmed to be identical to the native [Tyr32]POI by coelution high-performance liquid chromatography analysis and by enzymatic analysis using the phenoloxidase inhibition assay. To determine the disulfide pairings within the peptides, a series of enzyme hydrolyses and partial reduction/alkylation steps were performed. Three cystine pairs (Cys11-Cys25, Cys18-Cys29, and Cys24-Cys36) were determined by identification of the resulting peptides. The disulfide pairings of the two adjacent Cys residues (Cys11-Cys25 and Cys24-Cys36) were unambiguously assigned by comparing the derived fragments with the two possible isomers synthesized through a novel disulfide-linking technique. The arrangement of the disulfide bridges in POI was found to be topologically identical to those found for several peptides within the inhibitor cystine knot structural family. Although these peptides share a low primary sequence homology and display a diversity of biological functions, they nonetheless share similarities in their cystine motifs and tertiary structure. The tertiary structure model of POI, which was derived through molecular dynamics and energy minimization studies using restraints with determined disulfide connectivities, suggests that POI is a new class member of the inhibitor cystine-knot structural family.

Amino Acid Sequence↗