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Biomedical subjects

D C Chan

Publications and source records attributed to D C Chan.

At least 91 records · Page 5Linked to original sources
PubMed1989

Intrahepatic assembly of very low density lipoproteins: immunologic characterization of apolipoprotein B in lipoproteins and hepatic membrane fractions and its intracellular distribution.

Monoclonal antibodies, prepared against rat apoB, were used to examine apoB structure in serum lipoproteins and characterize the forms and localization of apoB in liver membrane fractions and cultured hepatocytes. Of the several antibodies obtained, four, having separate epitopes, were characterized. Western blot analysis showed that three (DB11, F4, and LB14) antibodies recognized both apoBL and apoBS. One antibody (HB41) recognized only apoBL. This antibody showed unusual properties. Competition ELISA assays showed that the epitope recognized by HB41 was more effectively expressed on low density lipoproteins (LDL) compared to very low density lipoproteins (VLDL). In addition, treatment of lipoproteins with detergents and sulfhydryl reducing agents also increased the expression of the HB41 epitope. Since HB41 has been found to inhibit LDL binding to hepatocyte receptors, these data indicate that the HB41 epitope is located on the carboxy-terminal side of the apoBS junction (probably within the LDL receptor binding domain). Western blotting hepatic microsomal subfractions showed that in the rough and smooth microsomes, HB41 recognized only apoBL, while in the Golgi it recognized both apoBL and a protein having a molecular weight slightly smaller. In contrast, Western blotting with a polyclonal antibody known to recognize both apoBL and apoBS showed that, in rough and smooth microsomes, proteins in addition to apoBL and apoBS having molecular weights between 120,000 and 30,000 were recognized. These proteins, likely to be proteolytic fragments of apoB, were barely detectable in the Golgi. Additional biosynthetic studies show that the [35S]methionine-labeled proteins smaller than apoB were immunoprecipitated from the rough microsome subfraction. Pulse-chase experiments show that these are produced with the same kinetics as full-size apoBL and apoBS, indicating that they are not incomplete nascent chains. Finally, immunofluorescence microscopy was used to determine the localization of monoclonal epitopes. ApoB monoclonal antibodies that recognized exclusively apoBL (HB41) and apoBL and apoBS (DB11) produced an immunofluorescence pattern characteristic of the endoplasmic reticulum, but not the Golgi. These data suggest that, in cultured rat hepatocytes, the majority of both molecular weight forms of apoB are localized in the endoplasmic reticulum, the initial site of VLDL assembly. The additional finding that proteolytic fragments of apoB are enriched in the microsomal fraction suggests that if the proteolysis occurs during subcellular fractionation, immature apoB is susceptible to proteolysis.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1985

Composite resin compatibility and bond longevity of a dentin bonding agent.

The shear strength of a commercial dentin bonding agent was studied using human dentin in vitro. The agent was tested for compatibility with three proprietary composite resins. In addition, some samples were subjected to prolonged water immersion and thermocycling to determine effects on bond strength. There was no statistically significant difference in shear strengths when the three composite resins were used. In addition, there was no difference in shear strengths among those samples thermocycled or immersed for up to one year. The implications of this study are that the dentin bonding agent may be used with several composite resins with no significant difference in bond strengths, and that the bond seems stable when subjected to immersion or simulated intra-oral thermal stresses.

Adhesives↗
PubMed1984

The role of the central globular domain of histone H5 in chromatin structure.

Histone H5 contains three tyrosines in the central, apolar region of the molecule. All three tyrosines can be spin labeled at low ionic strength. When the central globular domain is folded at high ionic strength, only one tyrosine becomes accessible to the imidazole spin label. Spin labeling the buried tyrosines prevents the folding of the globular structure, which, in turn, affects the proper binding of the H5 molecule to stripped chromatin. Chromatin complexes reconstituted from such an extensively modified H5 molecule show a weaker protection of the 168 base pair chromatosome during nuclease digestion. However, when only the surface tyrosine of the H5 molecule is labeled, such a molecule can still bind correctly to stripped chromatin, yielding a complex very similar to that of native chromatin. Our data supports the idea that not just the presence of the linker histone H5, but the presence of an intact H5 molecule with a folded, globular central domain in essential in the recognition of its specific binding sites on the nucleosomes. Our data also show that during the chromatin condensation process, the tumbling environment of the spin label attached to the surface tyrosine in the H5 molecule is not greatly hindered but remains partially mobile. This suggests that either the labeled domain of the H5 molecule is not directly involved in the condensation process or the formation of the higher-order chromatin structure does not result is a more viscous or tighter environment around the spin label. The folded globular domain of H5 molecule serves in stabilizing the nucleosome structure, as well as the higher-order chromatin structure.

Animals↗
PubMed1982

Effect of Tyrosyl modifications on nucleosome reconstitution: a spin-labeling study.

An imidazole spin-label was used to study the role of tyrosyl residues in the reassociation process for the nucleosome core particle. The nucleosome core particle, containing 145 base pairs of DNA and a histone core (two each of the four histones H2S, H2B, H3, and H4), was isolated from chicken erythrocytes. Native particles were first dissociated in 2 M NaCl and labeled with varying concentrations of imidazole spin-label. The labeled histone core and endogenous DNA were then reassociated back by salt step dialysis. Reconstituted spin-labeled complexes, purified by an isokinetic sucrose gradient, were found to have physical properties identical with those of unlabeled native particles. Spin-labeling the surface tyrosines of the histone core did not interfere with proper reassociation of the nucleosome core complex. ESR spectra of the reconstituted nucleosomes core complex are not the strongly anisotropic type, suggesting that labeled surface tyrosines in the histone core are not involved in specific DNA-histone interaction nor does wrapping of DNA on the histone core involve very close contact with the label. When labeling was carried out under denaturing conditions following exposure of the histone core to urea, additional histone tyrosine residues were spin-labeled. The resulting histone-DNA complexes that formed after reassociation had physical properties different from those of the native nucleosomes core. This result suggested that some of the "buried" tyrosines are essential for specific histone-histone interactions that lead to stable histone core structures. Spin-labeling the buried tyrosines prevented to compact supercoiling of DNA into nucleosome core particle.

Animals↗
PubMed1980

ESR spin label studies of the nucleosome core particle and histone core.

An imidazole spin label has been used to study the accessibility and conformational state of tyrosines in both the nucleosome core particles and histone core extracted from chicken erythrocytes. About 40% of the tyrosyl residues in the histone core can be labeled under nondenaturing conditions. However, less than 15% of the tryosyls in the nucleosome core particle can be labeled even at 200- to 300-fold M excess of label. The effect of urea on the conformational state of the spin-labeled tyrosyls in both the nuclesome core particles and the histone core has been studied. Ionic effects on the spin-labeled nucleosome core have been investigated. Several conformational transitions are observed in the range of 1 mM NaCl to 2.5 M NaCl. Three major transitions are found at 0.1 M to 0.6 M, 0.7 M to 1.8 M and 2 M to 2.5 M NaCl, respectively. The observed changes can be interpreted as swelling and conformational change of the inner histone core, gradual separation of DNA from the histone core, and tightening of the histone core.

Animals↗
PubMed1976

Conformational state of DNA in chromatin subunits. Circular dichroism, melting, and ethidium bromide binding analysis.

This study compares some physical properties of DNA in native chromatin and mono-, di-, trinucleosomes obtained after mild micrococcal nuclease digestion. Melting curves and derivatives are shown to be very similar from one sample to another although a shift from 79 to 82 degrees C is observed between the mainly monophasic peak of multimers and chromatin. Careful analysis of the positive band of the circular dichroism spectra shows the appearance of a shoulder at 275nm, the intensity of which increases from the mono- to the di- and trinucleosome. This shoulder is maximum for native chromatin. At the same time binding isotherms of ethidium - bromide are characterized by two highly fluorescent binding sites for all the samples but the product KN of the apparent binding constant of the higher affinity binding sites by the apparent number of those sites increases from the mono- to the di- and trinucleosome. There again the valus is maximum for native chromatin. Such results strongly suggest that the native state of chromatin requires something more than the indefinite repeat of an elementary subunit.

Animals↗
PubMedDate not supplied

Two-step provisional technique for onlay preparations.

Compared to full crown temporization, direct autocured resin provisional technique for intracoronal onlay preparations is more complicated and less predictable. Clinical concerns include possible pulpal irritation, extra chair time for contouring proximal and marginal excess, and a critically short time constraint to remove the curing resin in the "rubbery stage" to avoid distortion or "locking" into the preparation. A two-step direct technique using two new light-cured provisional resins addresses these problems. First, a light-cured, elastic resin is condensed against the internal preparation walls and confined within a wedged metal matrix band. Then, using a clear template matrix, the elastic foundation is overlaid with a strong and esthetic light-cured provisional resin.

Acrylic Resins↗
PubMedDate not supplied

Evaluation of different methods for cleaning and preparing occlusal fissures.

The effectiveness of different methods for cleaning and preparing occlusal fissures before placing sealants was evaluated. Extracted mandibular molars received such treatments as brushing, pumicing, bur preparing, and air abrasion before application of fissure sealants. FluroShield fissure sealant was then applied to the occlusal fissures. Specimens were subjected to thermo-cycling and then immersed in a 10% solution of methylene blue, and finally sectioned. The sections were examined and photographed in a stereomicroscope, and the dye penetration was recorded using a scoring system. The results indicated that only the control (brushing with a dry brush) and the pumicing groups demonstrated dye penetration to the base of the sealant. Teeth prepared with the #1/4 round bur and air abrasion demonstrated a better seal in evaluated fissures. For this study, those three groups (occlusal fissures prepared with the #1/4 round bur and two air abrasion methods), demonstrated significantly better sealing (P < 0.01) than the control group and the other groups tested.

Air Abrasion, Dental↗
PubMedDate not supplied

Custom matrix adaptation with elastic cords.

Proper placement of the matrix and wedge is critical to the success of proximal amalgam and posterior resin composite restorations. This paper has presented an easy technique for adaptation of the matrix in cases where the gingival cavosurface margin involves a concavity. Special case considerations were also discussed.

Composite Resins↗
PubMedDate not supplied

Effect of air/water rinse versus water only and of five rinse times on resin-to-etched-enamel shear bond strength.

This study compared the shear bond strength of composite resin bonded to etched, flattened enamel that had been rinsed for 0, 1, 2, 3, 5 or 20 seconds with either a water stream or an air/water spray. One hundred seventy-six molars were separated into mandibular and maxillary groups, then divided equally into 11 groups of 16 teeth each. The facial enamel surfaces were ground flat and etched for 20 seconds with 37% phosphoric acid gel. In one group, the etching gel was dried but not rinsed. In five groups the gel was rinsed with a direct stream of water at 22 psi. In the remaining five groups the gel was rinsed with an air/water spray (air at 53 psi, water at 22 psi). One tooth from each group was removed for scanning electron microscopy evaluation, leaving 15 specimens in each group for shear bond testing. Teflon tape with a 3 mm in diameter window was placed over each etched area, a light-curing liquid resin was applied and polymerized for 10 seconds, and a cylinder of light-curing composite resin was polymerized over the window. Specimens were thermocycled from 5 to 50 degrees C for 500 cycles (30-second dwell time). After 30 days' hydration at room temperature, the teeth were mounted and the composite resin cylinders were loaded in shear to failure with an Instron machine at a cross-head speed of 5 mm/min. Mean load to failure was calculated for each group. Specimens that were not rinsed had significantly (P < 0.001) less bond strength (mean 0.53 MPa) than any of the rinsed specimens (mean 18.7 MPa).(ABSTRACT TRUNCATED AT 250 WORDS)

Acid Etching, Dental↗