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Biomedical subjects

W G Wade

Publications and source records attributed to W G Wade.

At least 37 records · Page 2Linked to original sources
PubMed1998

Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA.

We report the design and evaluation of PCR primers 63f and 1387r for amplification of 16S rRNA genes from bacteria. Their specificity and efficacy were tested systematically with a bacterial species and environmental samples. They were found to be more useful for 16S rRNA gene amplification in ecological and systematic studies than PCR amplimers that are currently more generally used.

Bacteria↗
PubMed1997

The comparative effect of acidified sodium chlorite and chlorhexidine mouthrinses on plaque regrowth and salivary bacterial counts.

Acidified sodium chlorite (ASC) is recognised as a highly potent, broad spectrum antimicrobial system that has been successfully developed for uses in veterinary, food processing and medical device fields. The current studies aimed to investigate the persistence of antimicrobial action and plaque inhibitory properties of 3 ASC mouthrinses by comparison with positive control, chlorhexidine 0.12%, and placebo control, water, rinses. Both studies were randomised, double-blind, cross-over 5-cell designs balanced for carryover. The 1st study involved 15 healthy subjects who immediately before and at 30, 60, 180, 300 and 420 min after rinsing provided 2 ml saliva samples. The samples were immediately processed for total anaerobic bacterial counts recorded after 96 h incubation. Washout periods were a minimum of 3 days. The second study involved 20 healthy subjects who on day 1 of each study were rendered plaque free, suspended normal oral hygiene methods and commenced rinsing twice daily with the allocated rinse. On day 5, plaque was scored by index and area after disclosing with erythrosin. Washout periods were 2 1/2 days. The 3 ASC and chlorhexidine rinses produced similar reductions in salivary bacterial counts which remained significantly below the placebo control to 7 h. There were no significant differences between ASC and chlorhexidine rinses except at 30 and 60 min when significantly greater reductions were produced by 2 ASC rinses compared to the chlorhexidine rinse. Plaque indices and areas were considerably and significantly lower with the ASC and chlorhexidine rinses compared to the placebo rinse. There were no significant differences between plaque scores for the 3 ASC rinses and the chlorhexidine rinse, although for 2 ASC rinses plaque scores were lower than for the chlorhexidine rinse. The results indicate that the 3 ASC rinses have equivalent plaque inhibitory action to chlorhexidine as a rinse. Similar to chlorhexidine, the plaque inhibitory action of the rinses appears to be derived from a persistence of antimicrobial action in the mouth.

Anti-Infective Agents, Local↗
PubMed1996

Identification and discrimination of oral asaccharolytic Eubacterium spp. by pyrolysis mass spectrometry and artificial neural networks.

Curie-point pyrolysis mass spectra were obtained from 29 oral asaccharolytic Eubacterium strains and 6 abscess isolates previously identified as Peptostreptococcus heliotrinreducens. Pyrolysis mass spectrometry (PyMS) with cluster analysis was able to clarify the taxonomic position of this group of organisms. Artificial neural networks (ANNS) were then trained by supervised learning (with the back-propagation algorithm) to recognize the strains from their pyrolysis mass spectra; all Eubacterium strains were correctly identified, and the abscess isolates were identified as un-named Eubacterium taxon C2 and were distinct from the type strain of P. heliotrinreducens. These results demonstrate that the combination of PyMS and ANNs provides a rapid and accurate identification technique.

Bacterial Typing Techniques↗
PubMed1996

Phylogeny of oral asaccharolytic Eubacterium species determined by 16S ribosomal DNA sequence comparison and proposal of Eubacterium infirmum sp. nov. and Eubacterium tardum sp. nov.

16S rRNA gene sequences of Eubacterium brachy, Eubacterium nodatum, Eubacterium saphenum, Eubacterium timidum, and two previously unnamed taxa were determined. The results of a phylogenetic analysis indicated that all of the strains sequenced belonged to a deep branch of the low-G+C-content gram-positive group. The levels of 16S ribosomal DNA sequence similarity between species were low, suggesting that a number of genera may be represented in this group. The representatives of the two unnamed taxa, which were isolated from patients with periodontitis, were clearly distinct from the previously described species, and, therefore, the following two new species are proposed: Eubacterium infirmum (type strain, NCTC 12940) and Eubacterium tardum (type strain, NCTC 12941).

Base Sequence↗
PubMed1996

Rapid differentiation of Prevotella intermedia and P. nigrescens by 16S rDNA PCR-RFLP.

Prevotella intermedia and the newly described P. nigrescens cannot be reliably distinguished by phenotypic tests. In this study, restriction endonuclease digestion of amplified 16S rDNA (16S rDNA PCR-RFLP) was used to generate restriction profiles of the type strains of P. intermedia and P. nigrescens and 43 fresh isolates identified as belonging to one of the two species. Whole-cell protein profiles were obtained by SDS-PAGE for comparative purposes. The type strains of P. intermedia and P. nigrescens were easily distinguished by 16S rDNA PCR-RFLP and the fresh isolates were assigned to either species on the basis of their restriction profiles. The identifications obtained were identical to those obtained by protein profiles. 16S rDNA PCR-RFLP is a rapid and reliable method for the differentiation of P. intermedia and P. nigrescens.

Bacterial Proteins↗
PubMed1996

Molecular analysis of microflora associated with dentoalveolar abscesses.

The microflora associated with three dentoalveolar abscesses was determined by cultural and molecular methods. 16S rRNA genes were randomly amplified by means of conserved eubacterial primers and cloned. Restriction fragment length polymorphism analysis of the clones and amplified genes encoding 16S rRNA from the cultured bacteria was used to detect putative unculturable bacteria. Clones representative of five predominant groups of uncultured organisms were sequenced. Two were identified as Porphyromonas gingivalis and Prevotella oris, and one was found to be closely related to Peptostreptococcus micros. The remaining two clones did not correspond to known, previously sequenced organisms. One was related to Zoogloea ramigera, a species of aerobic waterborne organisms, while the other was distantly related to the genus Prevotella. This study has demonstrated the possibility of the characterization of microflora associated with human infection by molecular methods without the inherent biases of culture.

Adult↗
PubMed1995

Restriction fragment length polymorphism analysis of PCR-amplified 16S ribosomal DNA of human Capnocytophaga.

The confusion in the taxonomic status of the genus Capnocytophaga has made identification of strains and studies on the role of this genus in infectious diseases equivocal. In this study 33 strains of Capnocytophaga including reference strains and various clinical isolates, were studied using RFLP analysis of 16S ribosomal RNA genes. The 16S ribosomal RNA (rRNA) gene sequences from whole cell suspensions and isolated genomic DNA samples were amplified by the polymerase chain reaction (PCR) using eubacterial specific primers. PCR products were purified and characterized by single digestions with 12 restriction endonucleases. Five of these, BanI, CfoI, HaeIII, HphI and RsaII were found to discriminate reproducibly between strains, and restriction patterns (ribotypes) produced by these were analysed to clarify the classification of Capnocytophaga strains. Dendrograms inferring similarities were derived from these data by the UPGMA method. This analysis produced three major clusters of strains, each of which was associated with a previously proposed species type strain: C. gingivalis, C. sputigena and C. ochracea. The results support the division of Capnocytophaga into three species and demonstrate that, despite the heterogeneity of this genus, the modified ribotyping method provides a simple, rapid and reproducible way to identify Capnocytophaga strains.

Bacterial Typing Techniques↗
PubMed1995

Antimicrobial properties of delmopinol against oral bacteria.

Antimicrobial properties of the morpholine derivative delmopinol hydrochloride were investigated in vitro. Minimum inhibitory concentrations were measured for a range of oral bacteria by broth and agar dilution techniques and minimum bactericidal concentrations were measured in broth. The rate of kill was investigated by determination and analysis of kill curves. The results indicated that delmopinol is a relatively weak antimicrobial in comparison with chlorhexidine. However, it may selectively affect certain species of oral bacteria which are of importance in plaque formation.

Chlorhexidine↗
PubMed1995

A 6-month home-usage trial of 0.1% and 0.2% delmopinol mouthwashes (II). Effects on the plaque microflora.

The effects of 0.1% and 0.2% delmopinol mouthwashes on supragingival plaque flora were investigated in a 6-month home-use study. 141 subjects were studied from whom plaque was collected at baseline, 12, 24, and 36 weeks. Overall, there were no consistent effects on microscopic or total counts. However, there was a significant reduction in the proportion of dextran-producing streptococci in the active groups compared to the control group throughout treatment. There was no colonisation by Candida or Gram-negative aerobic bacilli in the active groups nor was there any decrease in susceptibility to delmopinol. Delmopinol appears to mediate its anti-plaque effect without causing a major shift in bacterial populations, although dextran-producing bacteria appear to be affected, which may have relevance to this agent's mode of action.

Adolescent↗
PubMed1995

Analysis of cultivable Porphyromonas gingivalis with trypsin-like protease enzyme activity and serum antibodies in chronic adult periodontitis.

OBJECTIVE: Trypsin-like protease (TLPase) enzyme produced by Porphyromonas gingivalis has been implicated as a virulence factor in the pathogenesis of periodontal disease. The aims of this study were to investigate the relationship between cultivable P. gingivalis, TLPase enzyme activity (BANA hydrolysis) and serum antibody levels against cell sonicate and a purified TLPase antigen from P. gingivalis W50. MATERIALS AND METHODS: Sub-gingival plaque samples were cultured for levels of P. gingivalis together with a chairside analysis of TLPase enzyme activity (Perioscan) from periodontitis and gingivitis sites of adult periodontitis patients. A TLPase from P. gingivalis was purified by gel filtration and ion exchange chromatography from the vesicle fraction for use as a test antigen. RESULTS: Elevated levels of P. gingivalis were found at periodontitis sites, however, there was no correlation with sub-gingival plaque TLPase enzyme activity. Adult periodontitis patients had higher levels of IgG and IgA against cell sonicate and TLPase antigens than did controls. Those patients who were P. gingivalis culture-positive demonstrated an elevated immune response against both cell sonicate and TLPase when compared to P. gingivalis culture-negative patients. Treatment resulted in an improvement of clinical indices and no cultivable P. gingivalis could be recovered from the treated sites and there was a concomitant decrease in IgG levels against the TLPase. There was no significant difference in BANA hydrolysis at gingivitis sites or periodontitis sites after treatment. CONCLUSIONS: Further longitudinal studies are suggested to investigate the role of the TLPase in the response to treatment of chronic adult periodontitis patients.

Adult↗
PubMed1994

An unclassified Eubacterium taxon in acute dento-alveolar abscess.

The microflora of pus samples aspirated from 50 acute dento-alveolar abscesses was examined. A total of 143 bacterial strains was isolated, consisting predominantly of Prevotella spp., alpha-haemolytic Streptococcus spp., Peptostreptococcus spp. and Eubacterium spp. An unclassified asaccharolytic Eubacterium taxon was encountered in 17 (34%) of the abscesses. This taxon was found to have a positive association with Fusobacterium spp. and a negative association with alpha-haemolytic Streptococcus spp.

Acute Disease↗
PubMed1994

The antibacterial and anti-staining properties of the novel anti-adherent agent M239, 144 alone and in combination with chlorhexidine.

The anti-adherent agent M239,144 has been shown to reduce the adhesion of S. sanguis strains to saliva-coated hydroxyapatite by 27-94%. The purpose of this study was to determine any antibacterial properties of M239,144 which might inhibit replication of the organisms which do adhere and to determine whether M239,144 interferes with the antibacterial properties of the antiseptic agent chlorhexidine. Maximum inhibitory dilutions (MIDs) were calculated using both broth and agar methods. 0.1% M239,144 displayed no antibacterial activity against S. oralis NCTC 7864 or S. sanguis NCTC 7863 using the broth dilution method and had no antibacterial activity against a range of plaque-forming organisms using the agar dilution method. The anti-bacterial activity of chlorhexidine against four of five oral plaque-forming species was reduced by 0.1% M239,144. The effect of a novel anti-adherent agent, MK239,144, on chlorhexidine staining was assessed in an in vitro model. 1% M239,144 completely prevented chlorhexidine staining at concentrations from 0.0002% to 0.2%. M239,144/chlorhexidine formulations show potential for a non-staining anti-plaque mouthwash.

Anti-Bacterial Agents↗
PubMed1993

Does gingivitis lead to periodontitis in young adults?

It has been assumed that poor oral hygiene predisposes to gingivitis with progression to periodontitis and tooth loss. However, a high prevalence of gingivitis occurs in some populations in which severe periodontitis is rare. To assess whether gingivitis is a reliable predictor of periodontitis, we compared the periodontal health of tea pickers and university students aged between 18 and 30 in Indonesia. Oral hygiene and gingival health was significantly better in students than in tea pickers. However, the prevalence and the severity of chronic periodontitis were similar in the two groups. Our findings suggest that gingivitis is a poor predictor of periodontitis in subjects younger than 30 years. Attempts to prevent periodontitis in young adults through antigingivitis measures such as plaque control may be unsuccessful.

Adolescent↗
PubMed1993

A 6-month home usage trial of a 1% chlorhexidine toothpaste (II). Effects on the oral microflora.

The effects of 1% chlorhexidine and 1% chlorhexidine/fluoride dentifrices on supragingival plaque flora were investigated in a 6-month home-usage study. 146 subjects were studied from whom plaque around 6 teeth was collected at baseline, 6 weeks and 24 weeks. Total anaerobic counts were reduced at 6 and 24 weeks in both active groups compared to control. No differences were seen in any other bacterial parameters. There was a significant decrease in the susceptibility to chlorhexidine of bacteria recovered from the subjects on the active treatments compared to control but, since the magnitude of effect was less than one dilution, this was not thought to be of any clinical significance.

Adolescent↗
PubMed1992

The effects of antimicrobial acrylic strips on the subgingival microflora in chronic periodontitis.

This study investigated the effects of root planing and/or the placement of acrylic strips containing chlorhexidine, metronidazole or tetracycline on the composition and antimicrobial susceptibility of the subgingival flora in chronic periodontitis. 101 periodontal pockets from 73 patients were entered into 6 treatment groups which were, chlorhexidine, metronidazole or tetracycline strips, root planing, root planing followed by metronidazole strips and a control, no treatment group. Total anaerobic counts and anaerobe/aerobe ratios were estimated from samples taken before treatment and 1, 2, 4, 8 and 12 weeks after treatment. In addition, a more detailed analysis of the effects of the treatments on the subgingival flora was carried out on 12 pockets in 12 patients. Tetracycline strips, metronidazole strips and root planing and metronidazole strips were more effective than chlorhexidine strips in causing reductions in total anaerobic count and anaerobe/aerobe ratio. However, the changes in microbial parameters rebounded to approach baseline levels 4 weeks after treatment. Chlorhexidine caused no detectable changes in the composition of the subgingival microflora, while metronidazole had a variable effect. Tetracycline appeared to effect major shifts in the composition of the microflora of treated pockets but caused a marked selection of tetracycline-resistant organisms.

Acrylic Resins↗