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Biomedical subjects

D Beighton

Publications and source records attributed to D Beighton.

At least 91 records · Page 5Linked to original sources

Marginal ditching and staining as a predictor of secondary caries around amalgam restorations: a clinical and microbiological study.

Caries at the margins of restorations is difficult to diagnose. In particular, the relevance of both marginal ditching and staining around amalgam restorations is unclear. This clinical study questions the relevance of marginal ditching and color change to the level of infection of the dentin beneath the margins of amalgam restorations. Clinically visible sites (330) on the tooth/restoration margin were selected on 175 teeth. The enamel adjacent to each site was noted as stained (a grey discoloration) or stain-free. One hundred and seventy-eight sites were clinically intact, 83 sites had narrow ditches (< 0.4 mm), and at 49 sites, wide ditches were present (> 0.4 mm). Twenty sites with frankly carious lesions were also included. Plaque was sampled at the tooth-restoration margin, and the dentin was sampled at the enamel-dentin junction below each site. Samples were vortexed, diluted, and cultured for total anaerobic counts, mutans streptococci, lactobacilli, and yeasts. Plaque samples showed that margins with wide ditches (> 0.4 mm) harbored significantly more bacteria, mutans streptococci, and lactobacilli than did clinically intact margins and margins with narrow ditches. There were no significant differences in the degree of infection of the dentin beneath clinically intact restorations and those with narrow ditches, but samples associated with wide ditches and carious lesions yielded significantly more bacteria, mutans streptococci, and lactobacilli. The color of the enamel adjacent to the sample site was irrelevant to the level of infection of the dentin beneath the filling margin, provided a frankly carious lesion was not present.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Diagnosis of secondary caries: a laboratory study.

Secondary caries is difficult to diagnose accurately. The purpose of this laboratory study was to investigate various non-invasive clinical and radiographic criteria which might predict the presence of carious dentine beneath the margin of the filling. A total of 331 sites, each 3 mm in length on the tooth restoration margin, were selected on 112 extracted and filled teeth. Thirty of these sites showed obvious carious cavities. Ditching was apparent in a further 70 sites, while 231 sites were clinically intact. Staining of the margin of the filling was recorded and radiographs were taken of posterior teeth. Restorations were then removed and the enamel-dentine junction (EDJ) immediately below the sites was examined for its consistency (hard/soft) and colour (stained/stain-free). Results showed that staining around a filling is not a reliable predictor of softening or discolouration of the dentine beneath. A clinical carious cavity and radiographic evidence of demineralisation indicate soft and discoloured dentine and should trigger operative intervention, while ditching alone should not.

Dental Caries↗

Identification of viridans streptococci associated with bacteraemia in neutropenic cancer patients.

Twenty-three viridans streptococcal isolates from pyrexial neutropenic patients with various malignant diseases were studied in a comprehensive identification scheme. Fourteen isolates were identified as Streptococcus oralis, five as S. mitis and two as S. salivarius but the remaining two could not be identified reliably. The virulence mechanisms associated with the ability of these species to survive and grow in vivo require further investigation but may involve the production of specific glycosidase and proteolytic enzyme activities.

Bacteremia↗

Production of specific glycosidase activities by Streptococcus intermedius strain UNS35 grown in the presence of mucin.

An isolate of Streptococcus intermedius from a brain abscess showed neuraminidase (sialidase), beta-D-galactosidase, N-acetyl-beta-D-glucosaminidase and N-acetyl-beta-D-galactosaminidase activities. The optimal pH values of these enzymes were 5.5-6.0, 5.5-6.0, 5.0-5.5 and 5.0-5.5, respectively. The km of the enzymes varied according to whether the type of substrate was chromogenic or fluorogenic; sialidase was most active at the lowest substrate concentrations, with a km of 0.01 mM. In semi-defined medium, with porcine gastric mucin--a model glycoprotein--as the sole source of fermentable carbohydrate, levels of the glycosidases were significantly increased. Addition of glucose to the mucin-containing medium, or growth of cells in media supplemented with glucose alone, repressed glycosidic activities and the majority of these were cell-associated. S. intermedius cells from cultures grown with mucin were able, simultaneously, to transport via sugar:phosphoenolpyruvate phosphotransferase (PTS) systems, monosaccharides which are constituents of carbohydrate side chains of glycoproteins. These cells also possessed significant levels of neuraminate-pyruvate lyase, involved in the intracellular catabolism of neuraminic acid; this was absent from cells grown with glucose. These mechanisms, collectively, may facilitate the persistence and growth of S. intermedius in vivo.

Brain Abscess↗

Degradation of hyaluronate by Streptococcus intermedius strain UNS 35.

Streptococcus intermedius strain UNS 35, a brain abscess isolate, produced extracellular hyaluronidase when grown in brain heart infusion broth. Chemical assays with this enzyme indicated that hyaluronate depolymerisation resulted in the formation of carbohydrate moieties with N-acetylglucosamine at the reducing terminal and containing an unsaturated carbon-carbon double bond. The nature of the products of this hyaluronidase were investigated further by high-field (400 MHz) proton (1H) NMR spectroscopy. Treatment of hyaluronate with the enzyme resulted in a series of new, sharp resonances in spectra (acetamido methyl group singlets located at 2.03 and 2.07 ppm, sugar ring proton multiplets in the 3.5-4.2 ppm chemical shift range, and doublets at 5.16 and 5.87 ppm) characteristic of low-M(r) oligosaccharide species, predominantly those containing glucuronosyl residues with delta 4,5-carbon-carbon double bonds. Comparison of spectra acquired from hyaluronidase-treated samples with that of an authentic sample of 4-deoxy-L-threo-hex-4-enopyranosyluronic-acid-N-acetylglucosamine (delta UA GlcNAc) indicated that this disaccharide was a major product arising from the actions of this enzyme. When used in minimal media, hyaluronate supported growth of S. intermedius, with lactate as the major metabolic end-product.

Carbohydrate Sequence↗

Metabolic cooperation in oral microbial communities during growth on mucin.

Hog gastric mucin has been used as a model glycoprotein to determine the role of particular glycosidases produced by different oral bacteria in the development of stable, diverse microbial communities. The patterns of glycosidase and protease activity were determined in pure cultures of ten representative species of oral bacteria using synthetic substrates. A five-member mixed culture was established in a chemostat, comprising species with minimal glycosidase and protease activity, in which hog gastric mucin was the major carbon and energy source. Introduction of additional species with novel enzyme activities (e.g. sialidase, alpha-fucosidase and endopeptidase) led to their establishment within the community to make communities with seven, eight, nine and ten members and resulted in an increase in the total viable counts of the microbial consortium. This increase in viable count was made up of the numbers of the newly added species as well as from a rise in the numbers of the existing community members. This result suggested that glycoprotein catabolism involved the synergistic and concerted action of several species with overlapping patterns of enzyme activity. Such metabolic cooperation results in the liberation of additional nutrients, and this may help to maintain the characteristic diversity of resident microbial communities found in many natural habitats.

Animals↗

A comparison of primary root caries lesions classified according to colour.

Three hundred and ninety-five primary root caries lesions in 117 patients were included in this study. Each lesion was clinically characterised (colour, dimensions, distance from gingival margin and texture) and a sample of infected or altered dentine was taken using a standardised sampling procedure. The relationships between the colour and the microflora, texture, physical dimensions and distance from the gingival margin of the lesions were investigated. Lesions in each colour group (light brown, yellow, dark brown and black) were identified in each texture category (soft, leathery and hard), except for black lesions which were recorded as only soft or leathery. Black soft and black leathery lesions had a significantly greater area and harboured more lactobacilli than all other types of lesions while black leathery lesions also harboured significantly more yeasts. There were no significant differences in the number of bacteria or mutans streptococci isolated from soft or leathery lesions irrespective of lesion colour. Overall, these data indicate that the colour of primary root caries lesions is not a reliable indicator of primary root caries activity. Assessment of the clinical severity of primary root caries lesions requires consideration of lesion texture and the distance of the lesion from the gingival margin rather than the colour of the lesion.

Actinomyces↗

Spectrophotometric method for the assay of glycosaminoglycans and glycosaminoglycan-depolymerizing enzymes.

Undegraded glycosaminoglycans form a complex with 1-ethyl-2-[3-(1-ethylnaphtho-[1,2-d]thiazolin-2-ylidene)-2- methylpropenyl]naphtho-[1,2-d]thiazolium bromide (Stains-all) in solution resulting in a characteristic shift in spectrum. Hyaluronic acid (a nonsulfated glycosaminoglycan) reacts with the dye to form a complex with an absorbance maximum at 650 nm, while sulfated glycosaminoglycans (chondroitin sulfates A and C, dermatan sulfate, keratan sulfate, and heparan sulfate) give rise to an increase in absorbance at 480 nm. Increases in absorbance at the appropriate wavelength are directly proportional to the concentration of glycosaminoglycan interacting with the dye. This phenomenon provided the basis for a sensitive spectrophotometric assay for the quantitative measurement of bacterial and mammalian glycosaminoglycan-depolymerizing enzymes. The basic assay method was adapted for use in 96-well microtiter trays, thus enabling large numbers of assays to be carried out simultaneously.

Biopolymers↗

The use of a caries detector dye during cavity preparation: a microbiological assessment.

During cavity preparation conventional tactile and optical criteria are used to assess the caries status of the enamel-dentine junction, cavity preparation being considered complete when this area is hard to a sharp probe and stain free. In the present study 201 cavities were prepared under rubber dam. When caries removal was considered complete using the conventional tactile and optical criteria, a caries detector dye (1% acid red in propylene glycol), which is claimed to stain 'infected' tissue red, was applied. Fifty-two per cent of cavities showed caries dye stain in some part of the enamel-dentine junction. Subsequent microbiological sampling of dye-stained and dye-unstained sites resulted in the recovery of low numbers of bacteria and revealed no difference in the level of infection of the two sites. It is concluded that the conventional tactile and optical criteria are satisfactory assessments of the caries status of tissue during cavity preparation and that subsequent use of a caries detector dye on hard and stain-free dentine will result in unnecessary tissue removal.

Adult↗

Copper plate replica SEM of developing dental plaque in monkeys (Macaca fascicularis).

The aim of the study was to examine the development of dental plaque in macaque monkeys (Macaca fascicularis). Copper plate replicas were constructed from impressions of the labial surface of one of the upper central incisor teeth after tooth cleaning and when plaque had accumulated for 12, 24 and 48 h in each of five animals. Scanning electron microscope examination of the replicas showed that bacteria were present on the tooth surface as scattered individual cells, which after 24 h had formed a continuous layer. Both coccoid and filamentous bacteria were visible in 48-hour-old plaque. Such a rapid accumulation of potentially periodontopathic dental plaque emphasizes the importance of regular and meticulous oral hygiene.

Animals↗

Relationships between mutans streptococci and perceived treatment need of primary root-caries lesions.

A total of 447 primary root-caries lesions from 169 dental patients was studied to determine the relationships between mutans streptococci and the perceived treatment need of primary root-caries lesions. Samples of this altered dentine for microbiological culture were obtained. Lesions were classified into 5 treatment categories: soft and restore, leathery and restore, leathery and debride of caries, leathery and treat chemotherapeutically, and hard, to receive no treatment. The total numbers of mutans streptococci decreased significantly with decreased treatment need. The percentage of mutans streptococci from lesions requiring no treatment was significantly less than from lesions requiring treatment. The frequency of isolation of mutans streptococci was significantly greater from lesions requiring more treatment. Significantly more lesions containing > 10(2) mutans streptococci were distributed in the groups with a greater perceived treatment need or with larger dimensions occluso-gingivally and/or mesio-distally or bucco-lingually or with a closer proximity to the gingival margin.

Adult↗

Relationships between yeasts and primary root-caries lesions.

A total of 447 primary root-caries lesions from 169 dental patients was studied to determine the relationships between their clinical severity and the number and frequency of isolation of yeasts. Yeasts were isolated more frequently from soft lesions, from lesions at the gingival margin and from lesions deemed to require restoration. These associations may be due to the aciduricity and acido- genicity of yeasts but the relatively low numbers found do not support a pathogenic role for yeasts in the aetiology of root caries: at best they may be marker organisms of the most severe disease.

Analysis of Variance↗

Effects of N-acetylglucosamine on carbohydrate fermentation by Streptococcus mutans NCTC 10449 and Streptococcus sobrinus SL-1.

We have investigated the ability of two species of streptococci isolated from the human oral cavity (Streptococcus mutans NCTC 10449 and Streptococcus sobrinus SL-1) to metabolize N-acetylglucosamine (GlcNAc), a naturally occurring amino sugar present in saliva and human glycoproteins, when provided as the sole fermentable carbohydrate and determined the effects of the presence of GlcNAc on the fermentation of other carbohydrates. S. mutans used GlcNAc at concentrations of up to 10 mM to increase cell numbers, but S. sobrinus was unable to ferment the amino sugar alone and its uptake only occurred in the presence of a fermentable carbohydrate. GlcNAc had a marked inhibitory effect on the ability of S. sobrinus to produce lactic acid from glucose, sucrose, and fructose, at the same time increasing the lag period and doubling time of batch-grown cells. Such patterns of inhibition were found with S. mutans, but the effects were less than those seen in S. sobrinus. In mixed culture studies of the two species, S. sobrinus became the predominant organism when 10 mM glucose was supplied as the sole fermentable carbohydrate, with a concomitant decrease in the numbers of S. mutans cells, but supplementation of the broth with 10 mM glucose and 10 mM GlcNAc resulted in the emergence of S. mutans as the predominant organism. S. mutans and S. sobrinus grown in media containing glucose possessed the ability to transport glucose and GlcNAc, probably via the same glucose-phosphotransferase system at similar rates. However, intracellular levels of N-acetylglucosamine-6-phosphate deacetylase and glucosamine-6-phosphate deaminase were markedly higher in S. mutans grown on glucose and GlcNAc than in S. sobrinus: 34 and 398 and 8 and 17 nmol of NADPH formed per mi per mg of protein for S. mutans and S. sobrinus, respectively. We propose that GlcNAc inhibited growth of S. sobrinus in media containing glucose and GlcNAc by competing with glucose for the glucose phosphotransferase, depleting intracellular levels of phosphoenolpyruvate, and possessing, in contrast to S. mutans, low levels of N-acetyl-glucosamine-6-phosphate deacetylase and glucosamine-6-phosphate deaminase activity. Together, these data suggest that in dental plaque, S. sobrinus when exposed to GlcNAc will have a reduced ability to compete with S. mutans for dietary carbohydrates, contributing to the greater frequency of isolation of S. mutans from human populations.

Acetylglucosamine↗

Chondroitin sulfate depolymerase and hyaluronidase activities of viridans streptococci determined by a sensitive spectrophotometric assay.

Sensitive spectrophotometric assays for the detection of bacterial chondroitin sulfate depolymerase and hyaluronidase activities were developed by using Stains-all (1-ethyl-2-[3-(1-ethylnaphtho-[1,2-d]thiazolin-2-ylidene)-2- methylpropenyl]naphtho-[1,2-d]thiazolium bromide). Stains-all interacts with hyaluronic acid to produce a shift in the absorption spectrum with a distinct absorption peak between 620 and 660 nm, while chondroitin sulfate interacts to form a distinct shoulder between 440 and 500 nm. Assays measure undegraded substrate. A collection of 110 strains of viridans streptococci, including representatives of all the currently recognized species, was studied. Streptococcus intermedius and S. constellatus degraded hyaluronic acid, while only strains of S. intermedius, primarily isolated from brain and liver abscesses, produced chondroitin sulfate depolymerase. S. intermedius, of all the viridans streptococci, produces the widest range of glycoprotein- and glycosoaminoglycan-degrading enzymes, which may contribute to the virulence of this species.

Chondroitinases and Chondroitin Lyases↗

Assessment of the clinical status of primary root carious lesions using an enzymic assay.

The microflora of root carious lesions (n = 151) and sound root surfaces (n = 30) in 22 institutionalized, older patients was investigated using a standardized method of sampling. Material was removed using a sterile excavator and the numbers of bacteria in each sample were determined using conventional culturing techniques and by rapid (2 h) fluorogenic enzyme assay. Correlation between bacterial counts and enzyme assay result was 0.873 (p < 0.001). The numbers of bacteria and fluorogenic enzyme assay values were significantly (p < 0.001) greater from soft lesions than from leathery lesions while hard lesions and sound root surfaces had significantly (p < 0.001) lower values than the other lesion types and were not different from each other. A similar trend was apparent when these values were correlated with treatment needs. The fluorogenic enzyme assay may provide a rapid, objective measure of root caries severity which might be used in the monitoring and comparison of treatment protocols.

Aged↗

Microbiological validation of assessments of caries activity during cavity preparation.

The operative management of primary and secondary caries assumes that all discoloured tissue at the enamel-dentine junction (EDJ) represents active disease and this is removed to arrest the carious process. This study aims to establish clinical criteria to differentiate between active and arrested caries at the EDJ using microbiological assessment of dentine samples to verify its clinical status. Radiographs were available for posterior teeth. Cavities (n = 205) were prepared under rubber dam. After gaining access, areas of the EDJ were chosen and assessments made of consistency (soft, medium, hard), colour (dark brown, mid-brown, pale) and moisture content (wet, dry). Dentine was removed by using a No. 3 round burr and placed in 1 ml of bacteriological culture broth. This sampling procedure was repeated at the same site once during cavity preparation and again when the cavity was judged as fully prepared. Samples were vortexed, diluted and cultured to give viable counts of the total anaerobic microflora, mutans streptococci and lactobacilli; viable counts were expressed as log10 (CFU per sample +1). Results showed no significant differences between the microflora of primary and secondary caries. The number of bacteria recovered diminished significantly as cavities were completed. Initial samples from soft and wet lesions harboured significantly more bacteria, lactobacilli and mutans streptococci than samples from medium, hard or dry lesions. Lesions visible on radiographs harboured more bacteria, including lactobacilli and mutans streptococci, while dentine colour was not discriminatory.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

A microbiological study of primary root-caries lesions with different treatment needs.

Samples of altered or carious dentin for microbiological culture were obtained from 301 primary root-caries lesions in 59 patients by means of a standardized sampling procedure. This involved the cleansing of each root surface of extraneous supragingival plaque by means of a hand-held toothbrush and distilled water and the collection of the sample with a sterile dental excavator passed through the entire vertical dimension of each lesion. The total number of colony-forming units (cfu) in each sample and the numbers of mutans streptococci (primarily Streptococcus mutans), lactobacilli, yeasts, and Gram-positive pleomorphic rods (GPPR) were determined. Individual bacterial counts were expressed as log10 (cfu per sample), as a percentage of the total number of bacteria per sample, and as a frequency of isolation from lesions with different clinical diagnostic criteria. Clinical measurements of each lesion were made for color, texture, position relative to the gingival margin, and treatment need. Lesions classified as soft yielded significantly more bacteria, mutans streptococci, lactobacilli, and GPPR than leathery lesions, which yielded more bacteria than hard lesions. Lesions were classified into 5 treatment categories: soft and restore, leathery and restore, leathery and debride of caries; leathery and treat therapeutically; and hard no treatment. The total numbers of bacteria, mutans streptococci, lactobacilli, GPPR, and yeasts decreased significantly with decreasing treatment need. The frequency of isolation of mutans streptococci, lactobacilli, and yeasts was significantly greater from lesions requiring restoration and from lesions situated within 1 mm of the gingival margin. We suggest that the majority of root caries is initiated adjacent to the gingival margin.

Actinomyces↗

Synergistic degradation of bovine serum albumin by mutans streptococci and other dental plaque bacteria.

Mutans streptococci (Streptococcus mutans and Streptococcus sobrinus) exhibited low levels of proteolytic activity against the model protein substrate, FITC-labelled bovine serum albumin, when incubated alone. Inclusion of other members of the dental plaque microflora in the assay usually resulted in marked increases in the degree of proteolysis and a high level of synergy. Interactions between mutans streptococci and either Streptococcus oralis or Fusobacterium nucleatum gave rise to the greatest degree of synergistic proteolytic degradation.

Dental Plaque↗