Biomedical subjects

Keiichi Yoshida

Publications and source records attributed to Keiichi Yoshida.

24 records · Page 2Linked to original sources

PubMed2003

Flexural fatigue strength of CAD/CAM composite material and dual-cured resin luting cements.

PURPOSE: To evaluate the flexural strength and flexural fatigue strength of a CAD/CAM composite and three resin luting cements. MATERIALS AND METHODS: Specimens of CAD/CAM composite (2 x 2 x 25 mm) were fabricated from a composite block (GN-I) using a low-speed cutting saw. Similar specimens of three dual-cured resin luting cements, Linkmax HV, Panavia Fluoro Cement, and RelyX ARC were polymerized in a laboratory-photo-curing unit (Triad II). Flexural strengths were measured at a cross-head speed of 1 mm/minute in 4-point bending (span = 20 mm; distance between inside loading points = 10 mm). The cyclic fatigue test was performed at 1.0 Hz on a fatigue tester. The staircase method was employed using a 10,000 cyclic load limitation, 5 MPa stress increment, and 20 specimens for each material. The initial test was performed at approximately 50% of the mean flexural strength. RESULTS: The flexural fatigue strength results followed the same relationship as those of the static flexural strength, and all mean values were statistically different between the four materials. The ratio of the fatigue strength to the flexural strength ranged from 0.5-0.7, with the dual-cured resin luting cements being lowest.

Analysis of Variance↗

PubMed2003

A clinical evaluation of the agar alginate combined impression: dimensional accuracy of dies by new master crown technique.

PURPOSE: To evaluate the dimensional accuracy of several impression methods including agar alginate combined impression in vivo; the marginal accuracy of stone dies was determined using a new electroformed master crown technique. MATERIALS AND METHODS: Cast cores with knife-edge and chamfer margins and electroformed master crowns were fabricated for 3 patients. Five impressions were taken of each preparation, using agar alginate combined impression and silicone impression materials. Dies were made after impression. The marginal fit of the master crown on each die was analyzed by four-way analysis of variance (ANOVA) and Tukey HSD test (p<0.05). RESULTS: The marginal fit of the master crown on the dies with chamfer margin was better than those with knife-edge margin for agar alginate combined impression. The shape of the margin did not affect the accuracy when silicone impression material was used. CONCLUSIONS: The results suggest that the agar alginate impression method is clinically acceptable for the chamfer margin, but shape of the margin may affect the dimensional accuracy of dies. The shape of the margin does not affect the accuracy of dies when silicone impression was used. Furthermore, the master crown made by electroforming technique could be useful for clinical evaluation of impression methods.

Agar↗

PubMed2002

Mapping the heparin-binding site on the 13-14F3 fragment of fibronectin.

Fibronectin, a multifunctional glycoprotein of the extracellular matrix, plays a major role in cell adhesion. Various studies have revealed that the human 13th and 14th fibronectin type III domains (labeled (13)F3 and (14)F3 here) contain a heparin-binding site. Mapping of the heparin-binding sites of (13-14)F3, (13)F3, and (14)F3 by NMR chemical shift perturbation, isothermal titration calorimetry, and molecular modeling show that (13)F3 provides the dominant heparin-binding site and that the residues involved are within the first 29 amino acids of (13)F3. Predictions from earlier biochemical and modeling studies as well as the x-ray structure of (12-14)F3 were tested. It was shown that the positively charged residues that project into the solvent from the ABE face of the triple-stranded beta sheet on (13)F3 are involved in binding, but (14)F3 does not appear to contribute significantly to heparin binding.

Amino Acid Sequence↗

PubMed2002

Characterization of a heparan sulfate octasaccharide that binds to herpes simplex virus type 1 glycoprotein D.

Herpes simplex virus type 1 utilizes cell surface heparan sulfate as receptors to infect target cells. The unique heparan sulfate saccharide sequence offers the binding site for viral envelope proteins and plays critical roles in assisting viral infections. A specific 3-O-sulfated heparan sulfate is known to facilitate the entry of herpes simplex virus 1 into cells. The 3-O-sulfated heparan sulfate is generated by the heparan sulfate d-glucosaminyl-3-O-sulfotransferase isoform 3 (3-OST-3), and it provides binding sites for viral glycoprotein D (gD). Here, we report the purification and structural characterization of an oligosaccharide that binds to gD. The isolated gD-binding site is an octasaccharide, and has a binding affinity to gD around 18 microm, as determined by affinity coelectrophoresis. The octasaccharide was prepared and purified from a heparan sulfate oligosaccharide library that was modified by purified 3-OST-3 enzyme. The molecular mass of the isolated octasaccharide was determined using both nanoelectrospray ionization mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry. The results from the sequence analysis suggest that the structure of the octasaccharide is a heptasulfated octasaccharide. The proposed structure of the octasaccharide is DeltaUA-GlcNS-IdoUA2S-GlcNAc-UA2S-GlcNS-IdoUA2S-GlcNH(2)3S6S. Given that the binding of 3-O-sulfated heparan sulfate to gD can mediate viral entry, our results provide structural information about heparan sulfate-assisted viral entry.

Animals↗

PubMed2002

Effect of hyaluronan oligosaccharides on the expression of heat shock protein 72.

We have previously shown that intraarticular treatment with a hyaluronan (HA) preparation (840 kDa), HA84, up-regulates heat shock protein 72 (Hsp72) expression and suppresses degeneration of synovial cells in an arthritis model. In that study, the HA84 administered was degraded into HA oligosaccharides in the synovial tissue, suggesting that HA84 or degradation products of HA may up-regulate Hsp72 expression. Thus, in the present study, we examined the effects of HA of various molecular sizes on Hsp72 expression and cell death in stressed cells. Western blotting analysis showed that treatment of K562 cells with HA tetrasaccharides up-regulated Hsp72 expression after exposure to hyperthermia. On the other hand, treatment of the cells with HA of other sizes (di-, hexa-, deca-, dodecasaccharides), HA84, or tetrasaccharides of keratan sulfate did not elicit any change in expression of the Hsp72 protein. Treatment of the cells with tetrasaccharides of HA up-regulated not only expression of the Hsp72 protein but also Hsp72 mRNA expression and enhanced activation of HSF1, a transcription factor controlling Hsp72 expression, after exposure to hyperthermia. Because the level of Hsp72 protein was not affected by tetrasaccharides of HA when the K562 cells were kept at 37 degrees C without any stress, it is evident that tetrasaccharides of HA did not act as a stress factor. In addition, tetrasaccharides of HA suppressed cell death in the case of K562 cells exposed to hyperthermia and of PC12 cells under serum deprivation. These results suggest that a certain size of oligosaccharides, i.e. the tetrasaccharides of HA, up-regulates Hsp72 expression by enhancing the activation of HSF1 under stress conditions and suppresses cell death.

Adjuvants, Immunologic↗

PubMed2002

Purification and characterization of heparinase that degrades both heparin and heparan sulfate from Bacillus circulans.

A heparinase that degrades both heparin and heparan sulfate (HS) was purified to homogeneity from the cell-free extract of Bacillus circulans HpT298. The purified enzyme had a single band on SDS-polyacrylamide gel electrophoresis with an estimated molecular mass of 111,000. The enzyme showed optimal activity at pH 7.5 and 45 degrees C, and its activity was stimulated in the presence of 5 mM CaCl2, BaCl2, or MgCl2. Analysis of substrate specificity and degraded disaccharides demonstrated that the enzyme acts on both heparin and HS, similar to heparinase II from Flavobacterium heparinum.

Bacillus↗