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Biomedical subjects

R W Bryant

Publications and source records attributed to R W Bryant.

At least 19 recordsLinked to original sources
PubMed2001

Development and validation of a competitive AKT serine/threonine kinase fluorescence polarization assay using a product-specific anti-phospho-serine antibody.

A competitive fluorescence polarization (FP) assay has been developed for the serine/threonine kinase, AKT. The FP assay has been formatted in a 384-well microtiter plate and automated using a pipeting workstation with performance suitable for high-throughput screening. The assay design utilizes a fluorescent phosphorylated peptide complexed to a product-specific anti-phospho-serine antibody. When unlabeled substrate is phosphorylated, by the kinase, the product competes with the fluorescent phosphorylated peptide for the antibody. The fluorescent phosphorylated peptide is then released from the antibody into solution resulting in a loss in polarization signal. Seven fluorescent phosphorylated peptides and 19 antibodies were evaluated for this assay. RARTSpSFAEPGK-Fl peptide and anti-phospho-GSK-3alpha Ser21 antibody gave the best affinity and change in polarization signal. The apparent kinetic constants were calculated for the FP assay and were consistent with reported values. The FP assay was validated with known inhibitors and the results compared to a radioactive Flashplate transfer assay, utilizing [(33)P]ATP and a biotinylated substrate, also developed in our laboratory. The IC(50) values generated were comparable between the two methods suggesting the competitive FP assay and Flashplate assay have similar sensitivities and abilities to identify inhibitors during screening.

Antibodies↗
PubMed1999

Compounds capable of generating singlet oxygen represent a source of artifactual data in scintillation proximity assays measuring phosphopeptide binding to SH2 domains.

We developed scintillation proximity assays (SPA) to discover compounds which inhibit phosphopeptide binding to Src homology 2 (SH2) domain proteins Grb2 and Syk. An assay artifact is reported here as a caveat to others. The SPA used an antibody to couple glutathione-S-transferase SH2 domain fusion proteins to scintillant beads coated with protein A. A pyrazoloquinolone and indolocarbazole inhibited [3H]phosphopeptide binding in both assays. Their potency in the SPA increased with prolonged (2 to 24 h) assay exposure to ambient light. They were inactive in absence of light and in an alternate binding assay. Both compounds absorbed visible light and generated singlet oxygen based on 2-methylfuran-trapping experiments. Their inhibitory activity was suppressed by the singlet oxygen scavengers sodium azide and dithiothreitol. The results suggest that compounds, not previously considered photosensitizers, generated enough singlet oxygen to damage oxidant-sensitive SPA components. Therefore, this SPA should be protected from light to minimize occurrence of false positives.

Adaptor Proteins, Signal Transducing↗
PubMed1998

A clinical evaluation of posterior composite resin restorations: 8-year findings.

OBJECTIVES: From a continuing investigation of the clinical performance of three different types of composite resin, the findings of the clinical evaluation at 8 years are presented. METHODS: One operator placed 330 restorations in Class I and Class II preparations in the posterior teeth of 72 patients. Every patient received at least one restoration of each type of material: a microfilled composite, a small particle hybrid, a relatively coarse particle hybrid, and an amalgam control. Restorations were evaluated using clinical criteria. RESULTS: Forty six patients attended the 8-year recall. Twenty-five of the 213 restorations (13.7% of the composites and 5.8% of the amalgams) originally placed in these 46 patients had previously failed or been lost from the study or were assessed as requiring replacement at the 8-year recall. Bulk fracture and secondary caries at the margin were the most common forms of failure in the composite restorations. Other failures or losses were associated with a non-margin defect in the composite, caries not associated with the restoration, pulpal considerations, extraction for orthodontics and reasons unknown. One-hundred and ninety-three restorations (including five that required replacement) were available for clinical evaluation at 8 years and these included 17 Class II restorations. Significantly fewer restorations placed with the coarse particle hybrid exhibited evidence of marginal deterioration. CONCLUSION: At 8 years, composite restorations in posterior teeth had failed at a rate two to three times that of amalgam restorations. The most common types of failure were bulk fracture and secondary caries at the margin.

Adolescent↗
PubMed1998

The influence of modified cavity design and finishing techniques on the clinical performance of amalgam restorations: a 2-year clinical study.

Use of modified cavo-surface angles (CSAs) and finishing techniques for amalgam restorations may influence the long term marginal fracture around these restorations. One operator placed 111 Tytin amalgam restorations in Class I and II preparations in the permanent teeth of 37 patients, each of whom received three restorations. One of these restorations was placed in a cavity prepared with a 'traditional' CSA and was finished more than 24 h after placement. Prepared cavities for the remaining two restorations for each patient were 'modified' with the aim of producing an occlusal CSA of 90-110 degrees. One restoration was carved-only and the other was finished more than 24 h after placement. Clinical evaluation of the restorations was carried out at baseline, 1 year and 2 years. At these times, the restorations were photographed and stone replicas were obtained from impressions taken at the cavity preparation stage to determine the cavo-surface angles. Careful application of the modified cavity design provided a mean cavo-surface angle of 109 degrees whereas a 'traditional' cavity design provided a mean cavo-surface angle of 126 degrees. Using the Mahler photographic method of evaluation, a combination of modification of the occlusal cavo-surface angle and finishing of the restoration was found to have an influence on the marginal fracture at 2 years. The clinical significance of these findings, with respect to the modified cavity design, is discussed.

Adolescent↗
PubMed1998

Three-year evaluation of computer-machined ceramic inlays: influence of luting agent.

OBJECTIVE: Changes in the occlusal marginal adaptation of computer-machined inlay restorations were assessed over 3 years. METHOD AND MATERIALS: Each of 21 patients received two Cerec (Vita Mark II) two- or three-surface restorations, with margins entirely in enamel, placed using two of four luting agents: a homogeneous microfilled resin composite, a fine hybrid composite, a relatively coarse hybrid composite, and an encapsulated glass-ionomer cement. Replicas of the restorations were made at baseline 6 months or 1 year (93% of original restorations), 2 years (79%), and 3 years (83%). The percentage of continuous margin on the occlusal margin was evaluated for the enamel-luting agent and luting agent-porcelain interfaces using a scanning electron microscope. RESULTS: Mean continuous margin values were high for all groups at the enamel-luting agent interface; there were no significant differences among the luting agents. For each luting agent, the percentage of continuous margin decreased significantly at the luting agent-porcelain interface over 3 years; differences among the luting agents at 3 years were not significant (Kruskal-Wallis test). After 3 years, the wear of the luting agents was significantly different. The least wear occurred with the microfilled resin composite; the coarse hybrid composite showed the most wear. CONCLUSION: The occlusal margins of Cerec inlays exhibited consistently high-quality adaptation at 3 years. The microfilled resin composite luting agent was more wear resistant than the hybrid composites and the glass-ionomer cement.

Bisphenol A-Glycidyl Methacrylate↗
PubMed1996

Microleakage of amalgam alloys: an update.

Using an in vitro air pressure test, the authors found a wide variation in the microleakage characteristics of amalgam restorations made from 33 recently manufactured precapsulated alloys. Spherical particle alloys as a group exhibited the highest microleakage values. When the authors changed the mercury/alloy ratios to produce 1 percent more mercury in the capsules by weight, microleakage decreased significantly. Because extensive microleakage of amalgam restorations has been related to increased postoperative sensitivity, the authors suggest that practitioners should be alert to this potential problem.

Air Pressure↗
PubMed1994

A clinical evaluation of posterior composite resin restorations.

There is a trend towards manufacturers seeking to provide dentists with a single, all-purpose composite resin, usually of the small particle hybrid type. This three-year clinical study compared the clinical performance of three different types of composite resin used in posterior teeth and identified several modes of failure. Of the 330 restorations (three composite resins and one amalgam control) initially placed in 72 patients, 223 restorations in 48 patients were available for evaluation at three years. Modified clinical criteria for assessing the restorations were able to discriminate among the composite resins. A microfilled composite and a small particle hybrid exhibited increasing evidence of marginal fracture (crevice) with time. In addition, the small particle hybrid showed evidence of wear at the margins more frequently than the other materials. Of the restorations available for assessment, four restorations of each of these two types of composite resin required replacement during the study. Coarse particle hybrid restorations showed evidence of wear but little evidence of marginal fracture.

Acrylic Resins↗
PubMed1993

The assessment of apical leakage of root-filled teeth by the electrochemical technique.

Since its inception by Jacobson and von Fraunhofer in 1976, several studies have used the electrochemical method to assess the leakage occurring around root canal fillings. This study has examined aspects of the electrochemical technique and the effects that this test method may have on leakage results. The findings have raised concerns about the application of the technique. It is evident that electrochemical leakage testing over 30 days can influence the electrochemical leakage reading at 30 days and may influence the linear dye penetration recorded at 30 days. It is suggested that testing for electrochemical leakage should not precede the assessment of linear dye penetration.

Bismuth↗
PubMed1992

Finishing of amalgam restorations: a three-year clinical study.

Class I and Class II amalgam restorations were placed in the permanent teeth of 66 patients, each of whom received a minimum of three restorations. At least one of these restorations was carved-only, at least one was immediately finished (using a prophylaxis paste on rubber cups at low speed, 8 or 10 min after trituration), and at least one restoration was polished (using finishing burs and polishing points, more than 24 h after placement). The number of restorations available for assessment of marginal fracture using the 11-unit photographic scale developed by Mahler was 253 at baseline, 249 at 1-year and 2-year recalls and 228 at the 3-year recall. Although polished restorations exhibited greater initial (baseline) marginal fracture, at 1-year, 2-year and 3-year recalls the restorations were found to exhibit a similar amount of marginal fracture regardless of the finishing technique. The amount of perceived marginal fracture increased steadily after 1 year. The immediate finishing of amalgam restorations showed no long-term benefit over the other techniques and has little to commend it. The clinical finishing of amalgam restorations is discussed in the light of this and other research.

Adolescent↗
PubMed1992

Direct posterior composite resin restorations: a review. 2. Clinical technique.

Despite the increased use of composite resin to restore posterior teeth, there is evidence that clinicians should be selective in their use of these restorations. This paper describes the clinical technique--preparatory procedures, preparation of the cavity, preparation for placement of composite resin, placement of composite resin and finishing of the restoration--for the relatively conservative use of composite resin in posterior teeth and reviews the literature to discuss briefly many of the controversial aspects of the technique.

Bicuspid↗
PubMed1992

Finishing techniques for amalgam restorations: clinical assessment at three years.

Use of the optimum finishing technique for an amalgam restoration may enhance the marginal integrity of the restoration and discourage its unnecessary early replacement. Two hundred and twenty-eight high copper amalgam restorations in 56 patients were evaluated, using clinical assessment criteria, up to three years after placement. Each patient had received at least one carved-only amalgam, at least one immediately finished restoration, and at least one amalgam that was polished at a subsequent appointment. Regardless of the finishing technique, the restorations exhibited similar marginal integrity up to three years after placement. Polished restorations were found to have substantially superior surface texture and less likelihood of surface discoloration. No evidence was found to support the use of immediate finishing techniques. The clinical significance of these findings, with respect to the need to polish amalgam restorations, is discussed.

Color↗
PubMed1992

Direct posterior composite resin restorations: a review. 1. Factors influencing case selection.

The principal factors that influence case selection for direct composite resin restorations in posterior teeth are discussed. These include the perceived preference for tooth-coloured aesthetics, survival rate and replacement of posterior composites, clinical problems and concerns such as occlusal surface defects and the numerous effects of the material's polymerization contraction, and the availability of alternative tooth-coloured techniques for restoring posterior teeth. Specific guidelines in case selection are suggested. A subsequent paper reviews controversial aspects of the restorative technique for posterior composites and, on this basis, defines important principles in the clinical technique.

Composite Resins↗
PubMed1992

The adaptation achieved by four root canal filling techniques as assessed by three methods.

Four root canal filling techniques--lateral condensation, McSpadden compaction, ultrasonic activation (Enac), and thermoplastic injection (Ultrafil)--were assessed for adaptation of the filling material to the canal wall. The adaptation and leakage were examined quantitatively using an electrochemical method and a linear dye penetration method, and qualitatively by radiographic evaluation. Using the electrochemical method, differences among obturation techniques were relatively small, although a greater proportion of the teeth that had been filled by lateral condensation exhibited no leakage. Radiographically, the techniques appeared to exhibit similar adaptation in the apical 6 mm. The techniques of lateral condensation and ultrasonic activation showed superior control of length of the root canal filling.

Bismuth↗
PubMed1992

The influence of modified cavity design and finishing techniques on the margin region of amalgam restorations: a simulated clinical study.

The use of high copper amalgams has substantially reduced the marginal fracture around amalgam restorations. Modified cavity designs and finishing techniques have been proposed for occlusal amalgam restorations in order to reduce further the long-term marginal fracture around these restorations. Clinical and laboratory studies are required before these techniques can be recommended to clinicians and taught to dental students with confidence. Using eighty extracted third molar teeth, this in vitro study examines four combinations of cavity design and finishing technique. After finishing, the restorations were photographed for evaluation of the initial marginal fracture attributable to the finishing technique, and sectioned twice to enable examination of the margin regions to determine the cavo-surface angles, amalgam margin angles and the occurrence of amalgam excess at and beyond the cavo-surface angle. Although careful application of the modified cavity design provided a mean cavo-surface angle of 100-110 degrees and assisted in achieving a mean amalgam margin angle of at least 70 degrees, long-term clinical and further laboratory studies are required before the cavity design modification can be recommended for routine use.

Analysis of Variance↗
PubMed1992

Actions of a 5-lipoxygenase inhibitor, Sch 40120, on acute inflammatory responses.

Sch 40120 (10-(3-chlorophenyl)-6,8,9,10-tetrahydrobenzo[b] [1,8]naphthyridin-5(7H)-one) is an inhibitor of the 5-lipoxygenase enzyme in rat neutrophils, human neutrophils and the the MC9 murine mast cell clone with IC50 values of 8, 4 and 7 microM, respectively. The drug was examined for its effects on acute inflammatory responses in the paw and pleural cavity of rats. The drug suppressed paw inflammation triggered by a reverse passive Arthus reaction or a subplantar injection of the polysaccharide carrageenan with p.o. ED50 values of 0.2 and 1.5 mg/kg, respectively. In reverse passive Arthus reaction and carrageenan pleurisy models, Sch 40120 was found to suppress both the cellular and fluid components of the acute inflammation. The p.o. ED50 values for inhibition of cells and fluid in pleurisy models were in the range of 0.1 to 0.7 mg/kg. When applied locally to the ears of mice, the drug blocked an arachidonic acid-induced and leukotriene-mediated ear inflammation with an ED50 of 0.072 mg/ear. These findings suggest that Sch 40120 is a potent anti-inflammatory agent that may be particularly useful in the treatment of inflammatory diseases such as psoriasis in which leukotrienes appear to be major mediators of the pathological symptoms that characterize the disease state.

Acute Disease↗
PubMed1991

Wheat germ agglutinin and other selected lectins increase synthesis of decay-accelerating factor in human endothelial cells.

Decay accelerating factor (DAF) is a cell-surface phosphatidylinositol-anchored protein that protects the cell from inadvertent complement attack by binding to and inactivating C3 and C5 convertases. We have measured DAF on human umbilical vein endothelial cells (HUVEC) by immunoradiometric assay after its removal by phosphatidylinositol-specific phospholipase C or Nonidet P-40 detergent extraction and have previously demonstrated that DAF synthesis can be stimulated by phorbol ester activation of protein kinase C. We now report that although stimulation (4-48 h) of HUVEC with various cytokines, including TNF, IL-1, and IFN-gamma, did not alter DAF levels, wheat germ agglutinin (WGA) (5-50 micrograms/ml), a lectin specific for binding N-acetyl neuraminic acid and N-acetyl glucosamine residues, increased DAF levels fivefold when incubated with HUVEC for 12 to 24 h. The lectins Con A and PHA also stimulated DAF expression twofold, whereas a number of others including Ulex europaeus, Bandeiraea simplicifolia lectin I, and Ricinus communis agglutinin I, which bind to endothelial cells, were inactive. The increase in DAF by WGA was inhibited by N-acetyl glucosamine (10-50 mM) but by neither N-acetyl neuraminic acid nor removal of surface N-acetyl neuraminic acid with neuraminidase. However, succinylated WGA, which has unaltered affinity for N-acetyl glucosamine but not longer binds N-acetyl neuraminic acid, was inactive. These data suggest that the binding of WGA to sugar residues alone is not sufficient to trigger DAF expression and that occupation of additional, specific sites are required. The increase in DAF levels on HUVEC was blocked by inhibitors of RNA and protein synthesis. We conclude that continuous occupation by WGA of specific binding sites on HUVEC triggers events leading to DAF synthesis. This unique, long term stimulation of endothelial cells by lectins may be relevant to cell:cell interactions at the endothelium.

Antigens, CD↗
PubMed1991

Severe abrasion of composite and amalgam restorations in a patient with lupus erythematosus systemic. Case report.

A case of repeated severe abrasion of amalgam and composite resin restorations in a patient under treatment for Lupus erythematosus systemic is reported. The patient had a history of audible nocturnal bruxism. Severe roughening occurred within three weeks of placement of the restorations and was related precisely to surfaces in contact with the opposing restored dentition. Examination by SEM revealed an unusual wear pattern of the amalgam restorations.

Adult↗