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Biomedical subjects

N M King

Publications and source records attributed to N M King.

At least 37 records · Page 2Linked to original sources
PubMed2001

In vitro replication and mutagenesis of a novel reversion vector with selective DNA damage in the supF gene.

The ability to detect the most common type of UV-induced mutation, the C to T transition, at the previously characterized hotspot at position 99 of the supF gene has been demonstrated in a selectively irradiated reversion vector, pLS189(Rev). The supF region was amplified, irradiated with 500J/m(2) UVC or unirradiated, and ligated into the pLS189(Rev) plasmid. A portion of ligated product plasmid containing the irradiated fragment was sensitive to nicking by T4 endonuclease V, indicating the presence of the most common type of UV-induced damage, the pyrimidine dimer. Plasmid containing the irradiated or unirradiated supF gene was replicated completely in cellular extracts from either HeLa or XP-A cells in vitro. Plasmid containing the irradiated supF gene showed an inhibition of total replication to a level similar to those of previous studies with plasmid molecules exposed in their entirety to 40J/m(2). Replication of selectively irradiated plasmid resulted in an average reversion frequency of 0.071% in the two extracts; a 42-fold increase over the average spontaneous reversion frequency of unirradiated plasmid. The reversion frequencies were not significantly different between extracts prepared from HeLa and XP-A cells, indicating that neither the repair status of the cell lines nor the XPA protein itself affect the frequency of C to T transitions at position 99 of the supF gene in plasmid replicated in vitro. These data indicate that the plasmid pSL189(Rev), containing the selectively UV-irradiated supF gene, is a useful and sensitive tool to study mutagenesis at a specific site. This approach may be applicable to the investigation of other environmental DNA-damaging agents, by allowing the target gene to be selectively damaged while maintaining the ability of the plasmid to replicate completely. Such a system, amenable to biochemical manipulation, may be very valuable in elucidating the function of novel proteins in the process of mutagenesis.

Base Sequence↗
PubMed2001

XPA protein alters the specificity of ultraviolet light-induced mutagenesis in vitro.

Studies of ultraviolet (UV) light mutagenesis have demonstrated mutations at common sites in the target genes of shuttle vector plasmids replicated in cultured cells or by cellular extracts. The reasons for the specific pattern of mutagenesis are largely unknown. We have examined the specificity of UV-induced mutagenesis by replicating plasmid pLS189, irradiated with 40 J/m(2) UVC or unirradiated, in either xeroderma pigmentosum group A (XP-A) or HeLa cellular extracts. The XP-A extract displayed slightly lower replication ability, but produced a higher mutant frequency, compared to that of HeLa extract. Use of irradiated plasmid inhibited replication by an average of 63% and increased the mutant frequency by an average of 16.7-fold. Analysis of mutation spectra revealed nonrandom patterns of mutagenesis that differed significantly between HeLa and XP-A extracts. In comparison to HeLa extract, replication in XP-A extract resulted in lower frequencies of GC --> AT transitions and tandem double-base substitutions, and a higher frequency of deletions. Replication in HeLa extract produced hotspots at positions 100, 108, and 156 that were not produced by XP-A extract. Furthermore, XP-A extract produced hotspots at positions 124, 133, and 164, sites not characteristic of previous UV-induced mutagenesis studies using XPA-expressing cells. Addition of purified XPA protein to reactions containing XP-A extract altered each of these parameters, including loss of the hotspots at positions 124 and 133, to yield a more HeLa-like spectrum. These results indicate a previously uncharacterized role of the XPA protein in influencing the specificity of UV-induced mutagenesis during DNA replication.

Base Sequence↗
PubMed2001

Adverse surface interactions between one-bottle light-cured adhesives and chemical-cured composites.

OBJECTIVES: This study examined the effect of one-bottle adhesives with different acidities on bonding to chemical-cured and light-cured resin composites. METHODS: Twenty-four non-carious human third molars were divided into eight groups. A flat dentin surface was created for each tooth. Acid-conditioned dentin surfaces were bonded with Prime&Bond NT (Dentsply), OptiBond SOLO (Kerr), Single Bond (3M) or One-Step (Bisco). Each adhesive group was covered with composite buildups, using either a light-cured (Z100, 3M) or a chemical-cured composite (BisFil 2, Bisco). Specimens were vertically sectioned into 0.9x0.9 mm beams. Microtensile bond strengths were recorded and failure modes were classified using a stereoscopical microscope. Four representative beams from each group were further prepared for SEM examination. RESULTS: Two-way ANOVA showed that the effect of adhesive types, composite curing modes and their interaction were statistically significant (P<0.001). Multiple comparison tests revealed no statistically significant difference in the bond strength of the four adhesives with the light-cured composite (P>0.05). However, they were significantly lower when used with the chemical-cured composite (P<0.01). A positive correlation was observed between the acidity of adhesives and the bond strengths of the chemical-cured composite. Failure occurred predominantly along the composite-adhesive interface, with microporosities on the adhesive surface and voids within the chemical-cured composite. SIGNIFICANCE: Air incorporated during mixing of chemical-cured composites only contributed partially to the decreased bond strength observed in simplified-step adhesives. Ultrastructural observations suggested the presence of a surface interaction between the uncured, acidic resin monomers from the oxygen inhibition layer of the adhesive and the initiator components in the chemical-cured composite.

Acrylic Resins↗
PubMed2001

The effect of fluoridated and non-fluoridated rewetting agents on in vitro recurrent caries.

OBJECTIVE: This study examined the in vitro caries inhibiting potential of fluoridated and non-fluoridated rewetting agents that are applied to acid-etched enamel and dentine before the use of a water-free, dentine adhesive. MATERIALS AND METHODS: Twelve caries-free premolars were divided into three groups of four teeth each. 2 x 3 x 1.5 mm cavities were prepared on the mesial and distal surfaces of each tooth, with half of the cavosurface margin in enamel and half in root dentine. In Group I (control), One-Step (Bisco, Schaumburg, USA) was applied without etching or rewetting agents. In Group II, cavities were acid-etched, rinsed, dried, and rewetted with Aqua-Prep (Bisco), a non-fluoridated rewetting agent, and then bonded with One-Step. Treatment for Group III was similar to Group II, except that Aqua-Prep F (Bisco), a fluoridated rewetting agent was used. Bonded cavities were restored with a non-fluoride-containing flowable composite (AEliteFlo, Bisco). Artificial carious lesions were induced in these specimens, from which multiple 100+/-20 microm thick longitudinal sections were prepared, yielding 16 specimens per group for evaluation with polarised light microscopy (PLM) and microradiography (MRG). Representative sections were processed for transmission electron microscopy (TEM) examination and scanning transmission electron microscopy/energy dispersive X-ray (STEM/EDX) analyses. RESULTS: The differences in demineralisation of dentine among the groups were not statistically significant for 'relative' lesion depth (p > 0.05, ANOVA, Student-Neuman-Keuls test), but highly significant for 'relative' lesion area (p < 0.001). Wall lesions were consistently present in Group I, while inhibition zones were invariably observed in Group III. 87.5% of Group II specimens exhibited neither wall lesion nor inhibition zone. TEM showed that remnant dentine apatite crystallites within the inhibition zones in Group III were larger and denser than those present within the corresponding wall lesions. STEM/EDX analyses confirmed the presence of calcium, phosphorus and fluorine in these plate-like crystallites. CONCLUSION: When used with a water-free, single-bottle dentine adhesive, a non-fluoridated rewetting agent is able to reduce, but cannot completely prevent recurrent caries. The use of a fluoridated rewetting agent is useful under the situation when microleakage occurs, by providing the additional benefit of fluoride-induced demineralisation inhibition.

Analysis of Variance↗
PubMed2001

Effect of delayed activation of light-cured resin composites on bonding of all-in-one adhesives.

PURPOSE: This study examined the effect of delayed activation of light-cured resin composites on the microtensile bond strengths of two all-in-one adhesives to sound dentin. MATERIALS AND METHODS: Flat dentin surfaces prepared from caries-free third molars were bonded with either Prompt L-Pop (ESPE) or One-Up Bond F (Tokuyama). Each adhesive was divided into 6 subgroups containing three teeth each. A hybrid composite was left to contact the cured adhesive surfaces for 0, 2.5, 5, 10 or 20 min before light activation. In the 6th subgroup, the cured adhesive was covered with a layer of nonacidic bonding resin before contacting the composite for 20 min. Composite-dentin beams, with average cross-sectional areas of 0.85 mm2 were prepared for microtensile bond testing. Representative fractured beams from the 0, 10- and 20-min delay subgroups were prepared for SEM examination. Additional specimens were prepared in the 0- and 20-min delay subgroups by replacing the hybrid composite with either a flowable composite or a composite liner to facilitate TEM preparation and examination. RESULTS: Kruskal-Wallis ANOVA and Dunn's multiple comparison tests showed that significant differences existed among the subgroups of each adhesive. Regression analyses showed that for each adhesive, there was an exponential decline in mean bond strength with increasing delay in light activation, and the correlation was highly significant (r = -0.99, p < 0.005 for Prompt L-Pop; r = -0.96, p < 0.01 for One-Up Bond F). SEM examination of fractured interfaces in the immediately light-cured subgroups revealed the absence of voids within the fractured composite. However, voids were consistently observed in the 10 min- and 20 min-delay subgroups. They corresponded to soapsuds-like blisters that were observed with TEM in the 20 min-delay subgroups. CONCLUSION: Adverse interactions between acidic adhesive resin monomers and tertiary amines in light-cured composites normally do not occur because of the fast rate of free-radical generation in photochemical redox reactions. However, such interactions can occur in all-in-one adhesives on prolonged contact of light-cured composites with the cured adhesive layer.

Composite Resins↗
PubMed2000

X-Ray crystal structure and molecular dynamics simulations of silver hake parvalbumin (Isoform B).

Parvalbumins constitute a class of calcium-binding proteins characterized by the presence of several helix-loop-helix (EF-hand) motifs. In a previous study (Revett SP, King G, Shabanowitz J, Hunt DF, Hartman KL, Laue TM, Nelson DJ, 1997, Protein Sci 7:2397-2408), we presented the sequence of the major parvalbumin isoform from the silver hake (Merluccius bilinearis) and presented spectroscopic and structural information on the excised "EF-hand" portion of the protein. In this study, the X-ray crystal structure of the silver hake major parvalbumin has been determined to high resolution, in the frozen state, using the molecular replacement method with the carp parvalbumin structure as a starting model. The crystals are orthorhombic, space group C2221, with a = 75.7 A, b = 80.7 A, and c = 42.1 A. Data were collected from a single crystal grown in 15% glycerol, which served as a cryoprotectant for flash freezing at -188 degrees C. The structure refined to a conventional R-value of 21% (free R 25%) for observed reflections in the range 8 to 1.65 A [1 > 2sigma(I)]. The refined model includes an acetylated amino terminus, 108 residues (characteristic of a beta parvalbumin lineage), 2 calcium ions, and 114 water molecules per protein molecule. The resulting structure was used in molecular dynamics (MD) simulations focused primarily on the dynamics of the ligands coordinating the Ca2+ ions in the CD and EF sites. MD simulations were performed on both the fully Ca2+ loaded protein and on a Ca2+ deficient variant, with Ca2+ only in the CD site. There was substantial agreement between the MD and X-ray results in addressing the issue of mobility of key residues in the calcium-binding sites, especially with regard to the side chain of Ser55 in the CD site and Asp92 in the EF site.

Animals↗
PubMed2000

Oral rehabilitation of a hypohidrotic ectodermal dysplasia patient: a 6-year follow-up.

This case report describes the oral rehabilitation of a female child with hypohidrotic ectodermal dysplasia over a 6-year time period. It demonstrates the need for periodic modification and replacement of a prosthesis, an orthodontic appliance, and a gingivoplasty. Although the initial treatment plan was considered to be a compromise due to limited cooperation, an improvement was observed in the patient's social behavior as a consequence of her dental treatment. The effects of unavoidable changes in the dental team over 6 years are also discussed.

Anodontia↗
PubMed2000

Mechanical disruption of dentin collagen fibrils during resin-dentin bond testing.

PURPOSE: To determine if collagen fibrils on the dentin side of failed resin-dentin interfaces undergo mechanical disruption during microtensile bond testing. MATERIALS AND METHODS: Extracted, caries-free human third molars were divided into four groups. The occlusal enamel was removed, leaving a flat dentin surface for bonding. Resin composite buildups were made after the acid-conditioned dentin was bonded with either Single Bond (S) or One-Step (O), and using either moist bonding (M) or air drying for 5 s (D). After storage in water for 24 h, the teeth were vertically sectioned into an array of 0.9 x 0.9 mm resin composite-dentin beams. They were stressed to failure using the nontrimming version of the microtensile bond test. Fractured dentin and resin composite sides of representative beams from each group that exhibited adhesive failures under light microscopy examination were prepared for scanning (SEM) and transmission electron microscopy (TEM). RESULTS: A two-way ANOVA showed that moist bond strengths were significantly higher than those made to dry dentin (M > D; p < 0.001), but that there was no difference between the adhesives (S vs O; p = 0.547). SEM analysis showed the presence of loose collagen fibrils within fractured hybrid layers in the dry groups, but not in the moist groups. TEM examination of the dry-bonded groups revealed collagen fibrils that were thinner and exhibited abnormally wide interfibrillar spaces when hybrid layers were intact. Within dry-bonded fractured hybrid layers, broad mechanical disruption zones could be seen, consisting of fibrils that were devoid of cross banding, defibrillation of the subfibrillar architecture, and gross disaggregation into microfibrils. In the moist-bonded groups, only short mechanical disruption zones were found along the torn edges of the collagen fibrils. The rest of the fibrils beyond the fracture site were intact and retained their periodicity. Mechanical testing of demineralized matrices yielded a maximum modulus of elasticity of 43.9 +/- 6.1 MPa. CONCLUSION: We speculate that adhesive resin has a protective function for demineralized collagen in well-infiltrated hybrid layers. We propose that both the collagen and resin contribute to load sharing during stress application until the final moment of rupture. On the other hand, collagen fibrils in poorly infiltrated hybrid layers, being unsupported by resin, undergo various degrees of irreversible mechanical disruption depending on how the stress is dissipated. The collagen fibril network has a much lower modulus of elasticity compared to those of resin-infiltrated fibrils or the demineralized dentin.

Acid Etching, Dental↗
PubMed2000

Bonding of a self-etching primer to non-carious cervical sclerotic dentin: interfacial ultrastructure and microtensile bond strength evaluation.

PURPOSE: The objectives of this study were 1) to examine the ultrastructural features of the resin-sclerotic dentin interface following the application of Clearfil Liner Bond II sigma to natural cervical wedge-shaped lesions, and 2) to evaluate the regional tensile bond strength of this self-etching primer at different locations on natural and artificially-created cervical lesions. MATERIALS AND METHODS: Deep cervical natural lesions were bonded using the self-etching primer. Micromorphology of the bonded interface at different locations within the lesions were examined using scanning electron microscopy (SEM), transmission electron microscopy (TEM) and scanning transmission electron microscopy/energy dispersive x-ray analysis (STEM/EDX). Ultrastructural features were further compared with the use of the same self-etching primer on artificial lesions created in sound cervical dentin. A nontrimming technique was used to evaluate the regional tensile bond strength from the occlusal, gingival, and the deepest central part of both natural and artificial cervical lesions. Beams with a mean area of 0.46 +/- 0.03 mm2 were prepared and were pulled to failure using a Bencor Multi-T testing device attached to an Instron universal tester. Bond strength results were evaluated using a two-way ANOVA design. RESULTS: A hypermineralized layer devoid of intact, banded collagen was invariably present on the surface of the natural lesions. Depending upon its thickness at different locations of the lesion, the action of a self-etching primer may be limited to this surface layer alone, producing a hybridized hypermineralized surface layer. Penetration of the self-etching primer into the underlying sclerotic dentin produced a hybridized complex containing a hybridized hypermineralized surface layer as well as a subsurface layer of hybridized intertubular dentin. Bacterial colonization of the lesion surface resulted in the formation of an additional zone of hybridized intermicrobial matrix over the surface of the lesions. Dentinal tubules remained blocked with sclerotic casts, and resin tags were rarely observed. Regional tensile bond strength results showed that the overall bond strength to natural sclerotic dentin was about 20% lower than sound cervical dentin, but was independent of the different locations within the lesions from which bond strength was evaluated. CONCLUSION: There were four factors that may have influenced the overall decrease in bond strength in natural cervical sclerotic lesions: a) the presence of a hybridized intermicrobial matrix together with entrapped bacteria may have weakened the bonds, b) inability of a self-etching primer to etch through a thick, hypermineralized surface layer, c) presence of a layer of possibly remineralized, denatured collagen at the base of the hypermineralized surface layer, and d) retention of acid-resistant sclerotic casts that obliterate the tubular lumina and prevent effective resin tag formation.

Acid Etching, Dental↗
PubMed1999

Rewriting the "points to consider": the ethical impact of guidance document language.

Human gene transfer research has been under way for nearly 10 years, with few clinically significant results to date; yet public and professional expectations still run high. Exaggerated expectations and misconceptions about gene transfer research are attributable in part to the terminology used to describe this research, which is misleading but, in the absence of more precise language, is commonly used. In particular, use of the term "gene therapy" to describe gene transfer research and use of the term "patient" in place of "subject" tend to cause both professionals and the public to confuse research with proven effective treatment. Investigators preparing to engage in human gene transfer research must use the NIH guidance document, "The Points to Consider in the Design and Submission of Protocols for the Transfer of Recombinant DNA Molecules into the Genome of One or More Human Subjects," written by the Recombinant DNA Advisory Committee, to prepare for the submission of a research protocol. Several corrections in the terminology employed by this guidance document could help to avoid misconceptions about gene transfer research and clarify both the promise and the limits of the research enterprise for investigators and subjects.

Ethics, Medical↗
PubMed1998

The oral health of children with clefts--a review.

A review of the studies of the caries prevalence and periodontal health of patients with cleft lip and palate (CLP) revealed that only five investigations of caries prevalence in CLP patients included children. One reported that CLP children did not have a higher caries prevalence in the permanent dentition, whereas more recent studies have reported a higher caries prevalence in both the primary and permanent dentitions of CLP children than in those of noncleft children. However, there is wide variation in the teeth examined and the method of presenting data on caries prevalence. Six papers have reported on the periodontal health of adult CLP patients, and only one has done so on that of children. The adult CLP patients had poorer oral hygiene and more gingivitis, but there is no conclusive evidence that they have a higher risk of developing periodontal disease. No data on the oral hygiene of CLP children were available, but it has been emphasized that they have significantly more gingivitis than noncleft children, especially in the maxillary anterior teeth.

Adolescent↗
PubMed1998

A catalogue of anomalies and traits of the permanent dentition of southern Chinese.

Variations in size, form and morphology of the teeth can result in anomalies such as macrodontia, microdontia, hyperdontia, hypodontia, double tooth, dens evaginatus, dens invaginatus, and talon cusp. A trait may occur more commonly in certain ethnic groups. Among these traits are shovel-shaped incisors, lingual tubercle, Carabelli's trait and protostylid of the molars. Because people are traveling more frequently than ever before, and there are many migrants in different parts of the world, it is increasingly important for dental professionals to be aware of the characteristics and prevalence of the anomalies and traits that occur in the different ethnic groups, so as to be able to make valid clinical judgments and to provide the appropriate treatment. The characteristics of these anomalies and traits are presented along with the prevalence figures in the southern Chinese.

Asian People↗