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Biomedical subjects

Keiichi Yoshida

Publications and source records attributed to Keiichi Yoshida.

At least 19 recordsLinked to original sources
PubMed2006

Bonding of dual-cured resin cement to zirconia ceramic using phosphate acid ester monomer and zirconate coupler.

This study evaluated the shear bond strength between dual-cured resin luting cement and pure zirconium (99.9%) and industrially manufactured yttrium-oxide-partially-stabilized zirconia ceramic, and the effect of MDP (10-methacryloyloxydecyl dihydrogen phosphate) primer (MP) and zirconate coupler (ZC) on bond strength. Two different-shaped pure zirconium and zirconia ceramic specimens were untreated or treated with various primers, including different concentrations of MP containing phosphoric acid ester monomer (MDP) in ethanol, ZC containing a zirconate coupling agent in ethanol, or a mixture of MP and ZC. The specimens were then cemented together with dual-cured resin luting cement (Clapearl DC). Half of the specimens were stored in water at 37 degrees C for 24 h and the other half were thermocycled 10,000 times before shear bond strength testing. The bond strengths of resin luting cement to both the zirconium and zirconia ceramic were enhanced by the application of most MPs, ZCs, and the mixtures of MP and ZC. For the group (MP2.0+ZC1.0) containing 2.0 wt % MP and 1.0 wt % ZC, no significant difference was observed between in shear bond strength before and after thermal cycling for both zirconium and zirconia ceramic (p > 0.05). For the other primers, statistically significant differences in shear bond strength before and after thermal cycling were observed (p < 0.05). The application of the mixture of MP and ZC (MP2.0+ZC1.0) was effective for bonding between zirconia ceramic and dual-cured resin luting cement. This primer may be clinically useful as an adhesive primer for zirconia ceramic restoration.

Biocompatible Materials↗
PubMed2006

Generation of the novel monoclonal antibody against TLS/EWS-CHOP chimeric oncoproteins that is applicable to one of the most sensitive assays for myxoid and round cell liposarcomas.

The fusion oncoproteins, TLS-CHOP and EWS-CHOP, are characteristic markers for myxoid and round cell liposarcomas (MLS/RCLS). Especially, the peptide sequence of 26 amino acids corresponding to the normally untranslated CHOP exon 2 and parts of exon 3 (5'-UTR) is a unique structure for these chimeric proteins. In this report, we have generated monoclonal antibodies against the unique peptide sequence of TLS/EWS-CHOP oncoproteins. These antibodies reacted with TLS-CHOP fusion protein, but not reacted with normal TLS and CHOP proteins by Western blot analysis. In addition, one of the antibodies also recognized the chimeric oncoprotein in archival paraffin-embedded tissue samples of MLS/RCLS. The oncoprotein was detectable by the antibody even in the paraffin-embedded tissue samples whose mRNAs were too degraded to be detected by a nested reverse transcription-polymerase chain reaction-based assay. Thus, the molecular assay using the novel antibody is expected to be one of the most sensitive diagnostic assays for MLS/RCLS.

Adult↗
PubMed2006

Hardness development of dual-cured resin cements through different thicknesses of ceramics.

This study investigated the Knoop hardness of a thin layer in three dual-cured resin cements (Linkmax HV, Nexus 2, and Variolink II HV) irradiated through or not through different thicknesses (1 through 5 mm) of a machinable ceramic. Hardness was recorded at a series of time intervals up to five days, starting from the end of a light irradiation period. Increase in hardness was more rapid over the first 0.5 hour; thereafter it continued at a low rate until maximum hardness was attained. Ceramic thickness had a significant influence on hardness in all dual-cured resin cements, especially when ceramic thickness was more than 4 mm. In addition, it was noted that the polymerization of Nexus 2 seemed to be more dependent on light exposure compared with the other two materials. Variolink II HV and Linkmax HV, on the other hand, seemed to indicate the potential of being compensated by chemical curing to some degree.

Analysis of Variance↗
PubMed2006

Study on the fracture strength of root reconstructed with post and core: alveolar bone resorbed case.

This study evaluated the influence of alveolar bone level on the fracture resistance of root restored with post and core. Forty-eight extracted human mandibular premolars were divided into six groups. Cast posts and cores were cemented (MN8 and MP8) or resin cores were built up with fiber posts and composite resin (FN8, FP8, FN4, and FP4). Post length was 8 mm (MN8, MP8, FN8, and FP8) or 4 mm (FN4 and FP4). Specimens were embedded 2 mm (MN8, FN8 and FN4) or 5 mm (MP8, FP8 and FP4) below cement-enamel junction. All specimens were loaded at 45 degrees to the long axis until fracture. With normal bone model, cast post and core (MN8) showed the highest fracture resistance (2262.4 N). However, in the resorbed bone model, there were no significant differences in fracture resistance between cast post and core and fiber post with composite resin.

Alveolar Bone Loss↗
PubMed2006

Shear bond strengths of four resin bonding systems to two silica-based machinable ceramic materials.

The purpose of this study was to evaluate the bond strength between four bonding systems (GC Ceramic Primer and Linkmax HV (CP+LM), Clapearl Bonding Agent and Clapearl DC (CBA+CL), Clearfil Mega Bond Porcelain Bonding kit and Panavia F2.0 (MB+PF), and RelyX Ceramic Primer and RelyX ARC (RC+RA)) and two machinable ceramics (Vitablocs Mark II, VMII and GN-I ceramic block, GNI). Shear bond strength was determined after 24-hour immersion in water or after thermocycling of 20,000 cycles. It was found that the post-thermocycling bond strength of each bonding system to VMII was significantly higher than that to GNI. VMII showed no significant differences between CP+LM, MB+PF, and CBA+CL--the values of which were higher than that of RC+RA. Regarding GNI, CP+LM showed the greatest bond strength after thermocycling among the four bonding systems used. It was concluded that the crystalline phase of the ceramics used might have an effect on bond strength.

Acrylic Resins↗
PubMed2006

Post-irradiation hardening of dual-cured and light-cured resin cements through machinable ceramics.

PURPOSE: To evaluate the surface hardness (Knoop Hardness Number) of the thin layer in three light-cured and dual-cured resin cements irradiated through or not through 2.0 mm thick machinable ceramics. METHODS: A piece of adhesive polyethylene tape with a circular hole was positioned on the surface of the ceramic plate to control the cement layer (approximately 50 microm). The cement paste was placed on the ceramic surface within the circle. The ceramic plate with resin cement paste was placed on a clear micro cover glass over a zirconia ceramic block to obtain a flat surface, and the material was polymerized using a visible-light-curing unit. The surface hardness was recorded at a series of time intervals up to 5 days, starting from the end of a light-irradiation period. RESULTS: The hardness steadily increased with post-irradiation time and tended towards a maximum, usually reached after 1 or 2 days. In all cases, the increase in hardness was relatively rapid over the first 30 minutes and continued at a lower rate thereafter. The dual-cured resin cement for each material showed a significantly higher hardness value than the light-cured resin cement irradiated either through or not through ceramics at all post-irradiation times. The resin cements cured through ceramic for each material were significantly less hard compared with those cured not through ceramics at all post-irradiation times.

Analysis of Variance↗
PubMed2005

Effects of 3-methylcholanthrene on the transcriptional activity and mRNA accumulation of the oncogene hWAPL.

hWAPL is a human oncogene associated with uterine cervical cancer. Here, we demonstrate that hWAPL transcription is induced by 3-methylcholanthrene (3-MC) in the cervical carcinoma-derived cell line SiHa. hWAPL transcription was analyzed with evaluation of the mRNA and heterogeneous nuclear RNA (hnRNA) levels by quantitative real time PCR analysis. Flow cytometric analysis suggested that the alteration of hWAPL mRNA levels is independent of cell cycle profile. We also found that DMSO and some components of FBS affect hWAPL transcription. Interestingly, when the aryl hydrocarbon receptor (AhR) function was inhibited by alpha-naphthoflavone (ANF), the induction of hWAPL transcription by 3-MC was greater than that in AhR-functioning normal cells. These observations suggest that there are complex mechanisms regulating the transcription of hWAPL. Furthermore, mRNA level of a mouse homolog of hWAPL in mouse uterus was induced by 3-MC injection into the abdominal cavity. Thus, some effects from 3-MC exposure on uterus may be mediated by the unscheduled overexpression of hWAPL.

Animals↗
PubMed2005

Novel heparan sulfate structures revealed by monoclonal antibodies.

The sulfated glycosaminoglycan heparan sulfate (HS) is found ubiquitously on cell surfaces, in the extracellular matrix, and intracellularly as HS proteoglycans. Because of the structural heterogeneity of HS, tissue-derived HS preparations represent a mixture of HS chains originating from different cell types and tissue loci. Monoclonal anti-HS antibodies have been employed to detect the localization of specific HS epitopes in tissues, but limited information has been available on the saccharide structures recognized by the antibodies. We have studied the saccharide epitope structures of four anti-HS antibodies, HepSS1, JM13, JM403, and 10E4, which all recognize distinct HS species as demonstrated by different patterns of immunoreactivity upon staining of embryonic rat and adult human tissues. The epitopes recognized by JM13 and HepSS1 were found almost exclusively in basement membrane HS, whereas JM403 and 10E4 reacted also with cell-associated HS species. The binding of HepSS1, JM403, and 10E4 to HS was dependent on the GlcN N-substitution of the polysaccharide rather than O-sulfation. HepSS1 thus interacted with N-sulfated HS domains, JM403 binding was critically dependent on N-unsubstituted GlcN residues, and 10E4 bound to "mixed" HS domains containing both N-acetylated and N-sulfated disaccharide units. By contrast, JM13 binding seemed to require the presence of 2-O-sulfated glucuronic acid residues.

Acetylation↗
PubMed2005

The keratan sulfate disaccharide Gal(6S03) beta1,4-GlcNAc(6S03) modulates interleukin 12 production by macrophages in murine Thy-1 type autoimmune disease.

It has been reported that disaccharides of the glycosaminoglycans (GAGs), heparin, or heparan sulfate suppress the production of cytokines. Therefore, we examined the effects of GAGs (keratan sulfate, hyaluronan, chondroitin, chondroitin sulfate, and heparin sulfate) disaccharides on production of interleukin (IL)-12, a pivotal cytokine in the Th-1 type immune system. Among the GAG disaccharides, only a keratan sulfate disaccharide, Gal(6-SO(3))-GlcNAc(6-SO(3)) (L4), suppressed IL-12 production in macrophages stimulated with lipopolysaccharides and interferon-gamma. Neither keratan sulfate chains nor keratan sulfate tetrasaccharides elicited any change in the IL-12 production. N-Acetyl-lactosamine, Gal-GlcNAc (LacNAc), also did not change IL-12 production. These results indicated that a certain size, i.e. disaccharide and sulfate, are essential to suppress IL-12 production. L4 was then applied to MRL-lpr/lpr mice, a Th-1 type autoimmune disease model. The treatment of MRL-lpr/lpr mice with L4 1) decreased in serum IL-12, 2) induced apoptosis in T cells in lymph nodes thereby suppressing lymphoaccumulation, and 3) suppressed hypergammaglobulinemia and glomerulonephritis. We showed previously that IL-12 suppresses cell death of T cells, thereby enhancing the lymphoaccumulation in MRL-lpr/lpr mice. Moreover, it has been reported that IL-12 deficiency in MRL-lpr/lpr mice diminishes lymphoaccumulation and delays glomerulonephritis. The treatment with L4 suppressed phosphoprotein kinase C and phosphoinositide 3-kinase expression in macrophages, suggesting that L4 suppresses IL-12 production by inhibiting phosphoprotein kinase C and phosphoinositide 3-kinase pathways.

Animals↗
PubMed2005

A dioxin sensitive gene, mammalian WAPL, is implicated in spermatogenesis.

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is an endocrine disruptor that produces a variety of toxic effects. We have isolated a mouse homolog of the hWAPL gene, termed mouse WAPL (mWAPL), as a target of TCDD by cDNA representational difference analysis from mouse embryonic stem cells. A statistically significant increase in mWAPL expression was observed at 0.1 microM TCDD in AhR-/- mouse embryonic fibroblast cells. Interestingly, at 1 microM TCDD, mWAPL mRNA levels decreased in AhR+/+ cells, but further increased in AhR-/- cells. hWAPL and mWAPL were highly expressed only in testes among normal tissue samples, and we observed mWAPL localization in the synaptonemal complex of testicular chromosomes. In addition, mouse testes decreased the expression of mWAPL mRNA after a single intraperitoneal injection of TCDD. Thus, mammalian WAPL such as hWAPL and mWAPL may be involved in spermatogenesis and be target genes mediating the reproductive toxicity induced by TCDD.

Amino Acid Sequence↗
PubMed2005

Detection of fusion genes in sarcomas using paraffin-embedded tissues.

Many sarcomas are characterized by specific recurrent chromosomal translocations resulting in gene fusions. The genes involved in almost all of these translocations have been cloned, greatly changing sarcoma diagnosis. At the biological level, these chromosomal translocations produce highly specific fusion genes that encode key molecules for tumor development. The clinical correlation between these translocation-derived genetic markers and discrete histopathological entities has been remarkable. Today, detection of fusion genes plays a crucial role in the diagnosis of sarcomas that harbor atypical clinical or pathological presentations. The focus of this brief review is the recent impact that cytogenetic and molecular detection of these translocations has had on sarcoma diagnosis using paraffin-embedded sections.

Animals↗
PubMed2005

The physical properties of a machinable resin composite for esthetic restorations.

To investigate the pre-clinical relevancy of a machinable composite, its physical properties were evaluated and compared with a machinable ceramic and two indirect composites. A machinable resin composite (GN-I composite, CO), a machinable ceramic (GN-I ceramic, CE), and two resin composites (Artglass dentin, AG; Estenia dentin, ET) were used. Compressive strength, diametral tensile strength, flexural strength, elastic modulus, and fracture load of standardized, premolar crown-shaped specimens were determined. In terms of compressive strength, diametral tensile strength, and flexural strength, AG showed significantly lower values than the other three materials. In terms of fracture load, specimens with 1.5 mm thick wall showed a higher value than those with 1.0 mm thick wall, and the value decreased in the order of ET, CE, CO, and AG. Marginal tipping was also observed in ET and CE. Within the limits of the current study, CO showed physical properties favorable for constructing esthetic restorations.

Analysis of Variance↗
PubMed2005

Bond strength between machinable glass-ceramic and dual-cured resin luting cements using silane coupling agents.

PURPOSE: To evaluate the shear bond strength of two dual-cured resin luting cements (Linkmax HV and Panavia Fluoro Cement) to machinable glass-ceramics (Decsy Machinable Ceramic) and the effect of three silane coupling agents (Clearfil Porcelain Activator, Ceramic Primer, and Porcelain Liner M) on the bond strength. METHODS: Disk-shaped specimens fabricated from machinable glass-ceramic blocks using a low-speed cutting saw were either treated or not treated with one of three silane coupling agents and then bonded together with one of two dual-cured resin luting cements. Specimens were stored in water at 37degrees C for 24 hours and/or thermal cycled 50,000 times before shear bond strength testing. RESULTS: Regardless of the resin luting cement and thermal cycling, specimens treated with the Clearfil Porcelain Activator showed the highest shear bond strength among all the treatments. Surface treatment of the Clearfil Porcelain Activator showed significantly greater shear bond strength after 50,000 thermocycles compared with other treatments. However, significant differences in bond strength were observed between 0 and 50,000 thermocycles for all treatments.

Analysis of Variance↗
PubMed2004

Alteration of chromosome positioning during adipocyte differentiation.

Chromosomes are highly restricted to specific chromosome territories within the interphase nucleus. The arrangement of chromosome territories is non-random, exhibiting a defined radial distribution as well as a preferential association with specific nuclear compartments, which indicates a functional role for chromosome-territory organization in the regulation of gene expression. In this report, we focus on changes in adipocyte differentiation that are related to a specific chromosomal translocation associated with liposarcoma tumorigenesis, t(12;16). We have examined the relative and radial positioning of the chromosome territories of human chromosomes 12 and 16 during adipocyte differentiation, and detected a close association between the territories of chromosomes 12 and 16 in differentiated adipocytes, an association not observed in preadipocytes. Although further studies are required to elucidate the underlying reasons for the adipocyte-specific translocation of chromosomes 12 and 16, our observations indicate that alteration of relative chromosome positioning might play a key role in the tumorigenesis of human liposarcomas. In addition, these results demonstrate the potential impact of higher order chromatin organization on the epigenetic mechanisms that control gene expression and gene silencing during cell differentiation.

Adipocytes↗
PubMed2004

Flexural fatigue behavior of machinable and light-activated hybrid composites for esthetic restorations.

The purpose of this study was to determine the flexural strength and flexural fatigue strength of a machinable composite (GN-I) and three hybrid composites (Artglass, Estenia, and Gradia). Specimens (2 x 2 x 25 mm) were polymerized in a laboratory photo-curing unit and then immersed in water at 37 degrees C for 24 h, 6 months, and 1 year. After each immersion period, flexural strengths (4-PFS) were measured at a cross-head speed of 1 mm/min in 4-point bending (span = 20 mm; distance between inside loading points = 10 mm). The cyclic fatigue test was performed at 2.0 Hz on a fatigue tester. The staircase method was employed for flexural fatigue strength (FFS) using a 5000 cyclic load limitation, 5-MPa stress increment, and 20 specimens for each material. Both 4-PFS and FFS of the machinable composite after all immersion periods were significantly greater than those of the three hybrid composites. The FFS results followed the same pattern as those of the 4-PFS. The Estenia material exhibited the highest 4-PFS and FFS after 24 h of immersion among three hybrid composites, whereas after 1 year of immersion, all three composites showed almost identical 4-PFS and FFS statistically. A strong correlation was observed between 4-PFS and FFS of the machinable composite and three hybrid resin composites.

Composite Resins↗
PubMed2004

Zirconate coupling agent for bonding resin luting cement to pure zirconium.

PURPOSE: To evaluate the shear bond strengths of two dual-cured resin luting cements to pure zirconium and the effect of zirconate coupling agent on the bond strength. METHODS: The two different-shaped pure zirconium specimens (99.9%) were untreated or treated with one of the four primers including zirconate coupler and then cemented together with one of the two dual-cured resin luting cements. Half of the specimens were stored in water at 37 degrees C for 24 hours and the other half thermocycled 20,000 times before shear bond strength testing. RESULTS: Regardless of the resin luting cement and thermocycling, specimens treated with the mixture of zirconate coupler and resin bonding agent showed the highest shear bond strength among the five treatments. Surface treatment with the mixture of zirconate coupler and resin bonding agent showed significantly greater shear bond strength compared with other treatments at 20,000 thermocycles. The application by the mixture of zirconate coupler and resin bonding agent on the pure zirconium metal surface appears to be effective for bonding between zirconium and dual-cured resin luting cements.

Aluminum Silicates↗
PubMed2003

Characterization of a heparan sulfate 3-O-sulfotransferase-5, an enzyme synthesizing a tetrasulfated disaccharide.

Heparan sulfate d-glucosaminyl 3-O-sulfotransferases (3-OSTs) catalyze the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to position 3 of the glucosamine residue of heparan sulfate and heparin. A sixth member of the human 3-OST family, named 3-OST-5, was recently reported (Xia, G., Chen, J., Tiwari, V., Ju, W., Li, J.-P., Malmstrom, A., Shukla, D., and Liu, J. (2002) J. Biol. Chem. 277, 37912-37919). In the present study, we cloned putative catalytic domain of the human 3-OST-5 and expressed it in insect cells as a soluble enzyme. Recombinant 3-OST-5 only exhibited sulfotransferase activity toward heparan sulfate and heparin. When incubated heparan sulfate with [35S]PAPS, the highest incorporation of35S was observed, and digestion of the product with a mixture of heparin lyases yielded two major35S-labeled disaccharides, which were determined as DeltaHexA-GlcN(NS,3S,6S) and DeltaHexA(2S)-GlcN(NS,3S) by further digestion with 2-sulfatase and degradation with mercuric acetate. However, when used heparin as acceptor, we identified a highly sulfated disaccharide unit as a major product. This had a structure of DeltaHexA(2S)-GlcN(NS,3S,6S). Quantitative real-time PCR analysis revealed that 3-OST-5 was highly expressed in fetal brain, followed by adult brain and spinal cord, and at very low or undetectable levels in the other tissues. Finally, we detected a tetrasulfated disaccharide unit in bovine intestinal heparan sulfate. To our knowledge, this is the first report to describe not only the natural occurrence of tetrasulfated disaccharide unit but also the enzymatic formation of this novel structure.

Amino Acid Sequence↗