Search PubMed⌕ Search

Biomedical subjects

M Ueda

Publications and source records attributed to M Ueda.

At least 145 records · Page 8Linked to original sources

Suicide gene therapy for human oral squamous cell carcinoma cell lines with adeno-associated virus vector.

The purpose of this study was to test the possibility of gene transfer as a new therapy for oral cancer. Adeno-associated virus (AAV) has already been used in the fields of cystic fibrosis and Parkinson's disease as a potential vector for gene therapy because of its wide host range, high transduction efficiency, and lack of cytopathogenicity. Four human oral squamous cell carcinoma cell lines were transduced with an AAV vector containing the beta-galactosidase gene (AAVlacZ) in vitro. Gene transduction efficiency was from 20 to 50% at a multiplicity of infection (MOI; for the purposes of this study the number of vector genomes per target cell) of 1x10(3), and nearly 100% of each cell line were transduced at an MOI of 1x10(4). Next, four cell lines were transduced with an AAV vector containing the herpes simplex virus thymidine kinase (HSVtk) gene, which sensitizes transduced cells to ganciclovir (GCV). Subsequent administration of GCV resulted in nearly 100% tumor cell killing at an MOI of 1x10(4) and from 70 to 80% tumor cell killing at an MOI of 1x10(3). These results suggest that AAV-mediated gene transfer of HSVtk and administration of GCV has potential as a new therapy for oral squamous cell carcinoma.

Antiviral Agents↗

Curative treatment of central hemangioma in the mandible by direct puncture and embolisation with n-butyl-cyanoacrylate (NBCA).

Management of central hemangioma in the mandible is difficult because of the abundant vascular network in this region. One of the most common signs of these patients, especially in the mixed dentition period, is hypermobility of the teeth with spontaneous hemorrhage from the surrounding gingival sulcus. Various therapeutic modalities have been considered, but surgery is the most frequently used. In cases of a large extensive lesion, however, intralesional injections of sclerosing agents have often been successful. A case of central hemangioma of the mandible with arteriovenous malformations in a 10-year-old girl is reported. She was treated with direct injection of an embolic material, n-butyl-cyanoacrylate, which brought satisfactory results. Preoperative embolisation of feeder vessels with Gelfoam and Avitene soaked in thrombin together with this direct injection is a safe treatment modality that is as effective as surgery.

Child↗

Convenient synthesis of aliphatic polyesters by distannoxane-catalyzed polycondensation.

A convenient synthesis of aliphatic polyesters by the distannoxane-catalyzed polycondensation of aliphatic dicarboxylic acids and aliphatic diols in solvents under azeotropic conditions was developed. To determine the optimum conditions for polycondensation of succinic acid with 1,4-butandiol, the effect of various types of catalyst and their concentration was investigated. By choosing the appropriate solvent, the polymerization was made to proceed in a two-phase system of solvent and molten polymer, and in the presence of 0.001 mol % of 1-chloro-3-hydroxy-1,1,3,3-tetrabutyldistannoxane (CHTD), poly(butylene succinate) (PBS) with a weight-average molecular weight of 277,000 was obtained. This method was then successfully applied to the synthesis of various aliphatic polyesters, such as poly(ethylene succinate), poly(1,4-cyclohexanedimethylene succinate), and poly(ethylene sebacate).

Biodegradation, Environmental↗

Characterization of the amino acid transport of new immortalized choroid plexus epithelial cell lines: a novel in vitro system for investigating transport functions at the blood-cerebrospinal fluid barrier.

PURPOSE: To establish and characterize a choroid plexus epithelial cell line (TR-CSFB) from a new type of transgenic rat harboring the temperature-sensitive simian virus 40 (ts SV 40) large T-antigen gene (Tg rat). METHODS: Choroid plexus epithelial cells were isolated from the Tg rat and cultured on a collagen-coated dish at 37 degrees C during the first period of 3 days. Cells were subsequently cultured at 33 degrees C to activate large T-antigen. At the third passage, cells were cloned by colony formation and isolated from other cells using a penicillin cup. RESULTS: Five immortalized cell lines of choroid plexus epithelial cells (TR-CSFB 1 approximately 5) were obtained from two Tg rats. These cell lines had a polygonal cell morphology, expressed the typical choroid plexus epithelial cell marker, transthyretin, and possessed Na+, K+-ATPase on their apical side. TR-CSFBs cells expressed a large T-antigen and grew well at 33 degrees C with a doubling-time of 35 approximately 40 hr. [3H]-L-Proline uptake by TR-CSFB cells took place in an Na+-dependent, ouabain-sensitive, energy-dependent, and concentration-dependent manner. It was also inhibited by alpha-methylaminoisobutylic acid, suggesting that system A for amino acids operates in TR-CSFB cells. When [3H]-L-proline uptake was measured using the Transwell device, the L-proline uptake rate following application to the apical side was five-fold greater than that following application to the basal side. In addition, both Na+-dependent and Na+-independent L-glutamic acid (L-Glu) uptake processes were present in TR-CSFB cells. CONCLUSIONS: Immortalized choroid plexus epithelial cell lines were successfully established from Tg rats and have the properties of choroid plexus epithelial cells, and amino acid transport activity was observed in vivo.

Amino Acids↗

Establishment of bone marrow-derived endothelial cell lines from ts-SV40 T-antigen gene transgenic rats.

PURPOSE: Postneonatal neovascularization is thought to result exclusively from the proliferation, migration, and remodeling of fully differentiated endothelial cells (ECs). Recently, it has been reported that bone marrow contains cells which can differentiate into ECs and contribute to neoangiogenesis in adult species. In this study, we tried to establish conditionally immortalized endothelial cell lines (TR-BME) derived from rat bone marrow. METHODS: Mononuclear cells were isolated and differentiated into ECs at 37 degrees C from the bone marrow of a transgenic rat harboring temperature-sensitive SV40 large T-antigen (ts T-Ag) gene. Then, the cells were transferred and incubated at 33 degrees C, a permissive temperature for ts T-Ag. Expression of vascular endothelial growth factor (VEGF) receptor (VEGFR)-1, 2, Tie-1, 2 and von Willebrand factor (VWF) were assayed by reverse transcriptase-mediated polymerase chain reaction (RT-PCR). RESULTS: We have established three cell lines incorporating 1,1'-dioctadecyl-3,3,3',3-tetramethylindo-carbocyanine perchlorate (DiI-Ac-LDL) with a spindle shape. One of these, clone 2, strongly expressed VEGFR-2, and weakly expressed VEGFR-1 and VWF. In contrast, clone 8 showed strong expression of Tie-1, 2, and VWF, and weak expression of VEGFR-1,2. All markers were expressed strongly in clone 3. CONCLUSIONS: These data confirm that the above three TR-BME cells are novel ECs derived from bone marrow progenitors.

Animals↗

The milk protein promoter is a useful tool for developing a rat with tolerance to a human protein.

Biopharmaceuticals intended for humans are immunogenic in animals. Antibodies associated with their administration make it difficult to perform repeated-dose pharmacology and toxicology studies in animals. Despite suggestions to solve this problem with transgenic animal technology, an effective strategy has not yet been reported. The objective of the present study was to provide an efficient strategy to develop rats tolerant to biopharmaceuticals such as human gene-based proteins. The present study used transgenic rat lines (lines 311-6, 308-5, and 305-1) carrying a fusion gene designed to express the human growth hormone (hGH) gene under the control of the bovine alphaS1 casein gene promoter. Three lactating females with the transgene, produced approximately 4 mg/ml, 300 microg/ml, and 10 ng/ml in their milk. Male 8-week-old rats from these three lines were immunized with hGH three times (week 0, 1, and 3 ) and the production of antibodies against hGH in their sera were examined at week 4. While the hGH serum antibody titers increased over 1000-fold in wild-type control rats, there was no detectable antibody against hGH in the sera of these three transgenic lines. Human growth hormone in their sera was undetectable (lines 308-5 and 305-1) or much lower than the endogenous biologic level of rat growth hormone (line 311-6). Importantly, lines 308-5 and 305-1 developed tolerance to hGH without detectable hGH in their sera and these lines will be very useful for the repeated dose pharmacology and toxicology studies. These results suggest that a milk protein promoter can be a useful tool to develop transgenic rats that are tolerant to biopharmaceuticals intended for humans.

Animals↗

Postnatal depression in Japanese mothers and the reconsideration of 'Satogaeri bunben'.

BACKGROUND: Postnatal depression occurs in 10-15% of Western women. In Japan, there is a traditional support system for perinatal women and there have been few prospective studies on postnatal depression in terms of cross-cultural studies. METHODS: First, a cross-cultural study on postnatal depression was performed. Ninety-eight Japanese women living in England and 88 Japanese women living in Japan were recruited and followed up to 3 months postnatally. The Edinburgh Postnatal Depression Scale (EPDS) and Schedule for Affective Disorder and Schizophrenia were used as a self-report questionnaire and psychiatric diagnostic interview. Diagnoses of depression were made by Research Diagnostic Criteria. Second, a study on 'Satogaeri bunben' (a traditional ritual (support system) for perinatal women in Japan; 'Satogaeri' means returning to the original family town or house and 'bunben' means delivery) was undertaken. Seventy-five mothers were asked to answer the questionnaire 6 months postnatally. The questionnaire was devised by the authors about the choice of Satogaeri bunben and the reasons for the decision. RESULTS: The incidence of postnatal depression was 12 and 17% in the English and Japanese groups of mothers, respectively. However, subjects from both groups supressed expression of their depressed mood in answering the EPDS. Satogaeri bunben itself did not lower the incidence of postnatal depression. In the study on Satogaeri bunben, 23 of 75 women did not choose Satogaeri bunben; moreover, seven of these women did not have good support from their own mothers. DISCUSSION: Having found that postnatal depression is not uncommon in Japanese women, a screening system should be developed, in particular because Japanese women would not naturally express their emotions, and Satogaeri bunben needs to be reconsidered qualitatively.

Adult↗

Mast cell chymase expression and mast cell phenotypes in human rejected kidneys.

BACKGROUND: Mast cells (MCs) are known to participate in various types of chronic disease, but their role in chronic renal rejection is poorly understood. Recently, distinct phenotypes of MCs have been described in humans by the demonstration of one protease, chymase. Hence, we questioned whether chymase in MCs could play a role in the pathogenesis of renal rejection in humans. METHODS: We investigated MC chymase expression and MC phenotypes, using immunohistochemical single- and double-staining techniques, in nephrectomy (N = 13) and biopsy (N = 8) specimens of human rejected kidneys. Tissue chymase levels were determined by enzymatic assay for chymase activity. We also examined the association between MC chymase expression and the degree of interstitial fibrosis in these renal allografts. RESULTS: Based on chymase positivity, rejected kidneys were divided into two groups, a chymase-negative [Chy(-)] group and a chymase-positive [Chy(+)] group. Quantitative analysis showed that the number of chymase-positive MCs and tissue chymase levels were significantly higher in the Chy(+) group than in the Chy(-) group. Furthermore, the interstitial fibrotic area in the Chy(+) group was significantly larger than that in the Chy(-) group. Immunodouble staining analysis also demonstrated that a new MC phenotype, positive for chymase but negative for tryptase, was present in the human rejected kidney. CONCLUSIONS: These results show that increased expression of chymase in MCs is related to the severity of interstitial fibrosis in human rejected kidneys.

Adolescent↗

Tissue engineering research in oral implant surgery.

In this article, we introduce some of the more extensively evaluated technologies using concepts of tissue engineering. We report on hard tissue engineering and soft tissue engineering and their utility for dental implant therapy. For hard tissue engineering, we evaluated human recombinant bone morphogenetic protein-2 and marrow mesenchymal stem cells using a model of sinus augmentation procedure in rabbit. We also describe distraction osteogenesis as another category for hard tissue engineering. In addition, we evaluate soft tissue management using cultured epithelial grafting for soft tissue engineering. The results of our tissue regeneration materials and methods in this study are positive. When the tissue engineering materials are used in clinics in the future, implant surgery could be the leading field.

Absorbable Implants↗

Donor leukocyte infusion for Japanese patients with relapsed leukemia after allogeneic bone marrow transplantation: indications and dose escalation.

To clarify the role of dose escalation of donor leukocyte infusion (DLI) in the treatment of relapsed leukemia after allogeneic bone marrow transplant (BMT), data from 100 patients were collected from 46 facilities in Japan and analyzed with respect to indications and infused cell dose. Complete remission (CR) was achieved in 11 of 12 (91%) patients with relapsed chronic myelogenous leukemia (CML) in the chronic phase, 3 of 11 (27%) with CML in the acute phase, 8 of 21 (38%) with acute myelogenous leukemia (AML), 6 of 23 (25%) with acute lymphoblastic leukemia (ALL), and 5 of 11 (45%) with myelodysplastic syndrome (MDS). The probability of remaining in CR at 3 years was 82% in CML patients in the chronic phase, but 0% in those with CML in the acute phase, 7% in those with AML, 0% with ALL, and 33% with MDS. Acute graft-versus-host disease (GVHD) (> or = 2) developed in 31 of 89 (34%) patients with human leukocyte antigen identical related donors and was fatal for 7 (7%). A leukocyte dose of 1 x 10(7)/kg of recipient body weight with CML in the chronic phase, 3 x 10(7)/kg of recipient body weight with MDS, and 1 x 10(8)/kg of recipient body weight with acute leukemia appeared to be optimal as an initial dose of DLI. However, the minimal dose of leukocyte developing fatal GVHD was 7 x 10(7)/kg of recipient body weight. These suggest that a relatively small dose of DLI ranging from 1 x 10(7)/kg to 5 x 10(7)/kg of recipient body weight should be administered initially then the infused escalating dose 2 or 3 months later in patients with CML in the chronic phase and MDS. However, a large number of leukocytes around 1 x 10(8)/kg are needed to induce graft versus leukemia effects in patients with acute leukemia despite a 7% fatality in GVHD.

Adolescent↗

The fate of developing teeth in mandibular lengthening by distraction: an experimental study.

Purpose: The purpose of this study was to observe developing teeth in a lengthened mandible after distraction. Material: Ten mongrel dogs with deciduous dentitions were used. Methods: A corticotomy was carefully made around a tooth bud and the external distractor (Orthofix M-100((R))) was connected. After a 5-day latent period, distraction was started at a rate of 0.75 mm per day for 10 consecutive days. Then, the lower jaw was stabilized by an external fixation to allow ossification. While the operation was performed on the left side (Distraction group), the contralateral side was studied for comparison (Control). In addition, a corticotomy, artificial fracture and external fixation were carried out to confirm the influence of the operation (Fracture group). Then macroscopic, radiographic and histological evaluations were carried out. Results: In the Distraction group, the space between the wall of the dental follicle and the crown expanded as distraction began. The end of the calcified root became wider and irregular during the distraction period, and finally, the apex closed. In the Fracture group, the teeth erupted although slight alterations of the root shape were observed in association with the operation period. Conclusion: The root became irregular, but the teeth erupted within the distraction area. Copyright 2001 European Association for Cranio-Maxillofacial Surgery.

Journal Article↗

MR imaging of the lingual thyroid. Comparison to other submucosal lesions.

PURPOSE: To study MR findings for clues to the diagnosis of lingual thyroid. MATERIAL AND METHODS: MR findings and clinical and scintigraphic data of 5 cases of lingual thyroid were reviewed and the MR findings were compared to those of 16 cases of other submucosal lesions in the base of the tongue. RESULTS: Four of the 5 patients with lingual thyroid were women and all had hypothyroidism. MR imaging depicted lingual thyroid in the midline in the base of the tongue (n=5) and additional ectopic thyroid glands in the floor of the mouth (n=2) or between the right and left sternohyoid muscles (n=1). Ectopic thyroid glands appeared isointense or hyperintense relative to muscle tissue on T1-weighted images and showed slight or fair contrast enhancement. All glands had low to intermediate T2 signal, which was also seen in 1 case of non-Hodgkin's lymphoma and 1 case of adenoid cystic carcinoma. All ectopic thyroid glands had well-defined margins, whereas malignant tumors tended to have ill-defined margins and to invade the surrounding structures. All but the 5 cases of lingual thyroid had an MR-demonstrable thyroid gland in the normal cervical position. CONCLUSION: A well-defined mass of low-intermediate T2 signal in the midline base of the tongue, neither with invasive tendency nor with a cervical thyroid gland in the normal site on MR imaging, may strongly indicate lingual thyroid.

Adolescent↗

Generation of cytotoxic T lymphocytes by MHC class I ligands fused to heat shock cognate protein 70.

Immunization with gp96 and heat shock cognate protein 70 (hsc70) purified with in vivo bound naturally occurring peptides or bound to synthetic peptides by in vitro reconstitution has been shown to induce peptide-specific cytotoxic T lymphocytes (CTL). In addition, mycobacterial heat shock protein 70 covalently fused to ovalbumin (OVA)-derived fragments has been shown to generate MHC class I-restricted CTL responses. Here, we genetically fused five different CTL epitopes, including peptides derived from Plasmodium yoelii circumsporozoite protein, tumor antigens, HY antigen and OVA, to either the N- or C-terminus of murine hsc70 and expressed the resulting proteins in Escherichia coli. Vaccination with all five fusion proteins induced peptide-specific CTL, indicating that no cognate flanking regions of CTL epitopes are necessary for the immune response. The point of injection was crucial for CTL induction. CD4(+) T cells were not required for the priming of CD8(+) T cells and vaccination with bone marrow-derived dendritic cells pulsed with hsc70 fusion proteins also elicited CTL responses. Furthermore, by using deletion mutants of hsc70, we identified amino acid residues 280-385 of hsc70 as the region most critical for inducing the CTL response.

Animals↗

Association of vitamin D receptor gene polymorphism with periodontal diseases in Japanese and Chinese.

We examined whether polymorphisms in the vitamin D receptor (VDR) gene are associated with the incidence of adult periodontitis (AP) and early-onset periodontitis (EOP) in case-controlled studies of Japanese and Chinese subjects. Restriction fragment length polymorphisms in the VDR gene detected by digestion with Taq I were found to be significantly associated with the occurrence of AP or EOP, suggesting that the VDR genotype a risk factor for periodontitis.

Adult↗