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Biomedical subjects

M F Maiden

Publications and source records attributed to M F Maiden.

At least 19 recordsLinked to original sources

Serum IgG reactivity to subgingival bacteria in initial periodontitis, gingivitis and healthy subjects.

BACKGROUND/AIMS: Established periodontal diseases may be associated with antibody responses to periodontal pathogens, but it is not known at which stage of disease this antibody response is initiated. This study aimed to characterize the host systemic response in initial periodontitis, gingivitis, and periodontal health, to evaluate whether elevated serum antibodies to subgingival species could be detected in initial periodontitis. METHOD: Human systemic immune response were evaluated to 40 subgingival bacterial species in 16 healthy, 21 gingivitis, 11 initial periodontitis and 5 progressing recession adults. Subjects had minimal periodontal attachment level (AL) loss at baseline. Disease categories were determined after 12 months monitoring at three-month intervals. Increased AL loss > or = 1.5 mm (disease activity) at interproximal sites defined initial periodontitis, recession was characterized by AL loss at buccal sites. Serum IgG antibodies were evaluated semi-quantitatively by immunoblot from blood taken at baseline, active and final visits. RESULTS: No antibody was detected from 55% of reactions. When detected, levels were below those reported for advanced periodontitis subjects. There were no major differences in serum antibody levels between healthy, gingivitis and initial periodontitis subjects, despite differences in the subgingival microbiota. Serum antibodies for more species were detected in recession subjects, compared with the other study subjects. No changes in antibody levels were detected between baseline, active, and final visits. No systematic association between species colonization and presence of systemic antibody was observed. CONCLUSIONS: This study did not detect differential elevation of mean serum antibody levels in initial periodontitis subjects, suggesting that serum antibody levels are not sensitive risk markers for initial periodontitis.

Adult↗

Pre- and post-implantation microbiota of the tongue, teeth, and newly placed implants.

OBJECTIVES: This investigation sought intra-oral sources of species colonizing dental implants. MATERIALS AND METHODS: Plaque samples were taken pre- and post-successful osseointegration from implants, teeth, and from tongues of 10 edentulous and 11 partially dentate subjects. Samples were assayed using whole genomic DNA probes in a checkerboard assay to 42 subgingival species. RESULTS: Similar prevalences and mean levels (10(3) to 10(4)) of microorganisms colonized implants and teeth. Species levels from tongue samples were higher than those of teeth and implants, although species prevalence was similar, suggesting that larger samples were obtained from the tongue. No significant differences were observed between the microbiota from the tongue of edentulous and partially dentate subjects. Most implant species were detected on tongue pre-implantation. In individual edentulous subjects, there were positive associations between Capnocytophaga ochracea and Campylobacter rectus from tongue and implant samples. In individual partially dentate subjects, there were positive associations between Fusobacterium nucleatum subsp. vincentii from tongue and implant samples, and Treponema denticola from implant and tooth samples taken at the same visit. CONCLUSION: This study indicated that the tongue, in addition to teeth, can be a source for species colonizing new implants.

Adult↗

Characterization of Bacteroides forsythus strains from cat and dog bite wounds in humans and comparison with monkey and human oral strains.

Bacteroides forsythus strains recovered from cat and dog bite wound infections in humans (n = 3), monkey oral strains (n = 3), and the human oral ATCC 43037 type strain were characterized by using phenotypic characteristics, enzymatic tests, whole cell fatty acid analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, PCR fingerprinting, and 16S rDNA (genes coding for rRNA) sequencing. All three bite wound isolates grew on brucella agar supplemented with 5% sheep blood, vitamin K1, and hemin. These strains, unlike the ATCC strain and previously described monkey oral and human clinical strains, did not require N-acetylmuramic acid supplementation for growth as pure cultures. However, their phenotypic characteristics, except for catalase production, were similar to those of previously identified strains. PCR fingerprinting analysis showed differences in band patterns from the ATCC strain. Also, SDS-PAGE and whole cell fatty acid analysis indicated that the dog and cat bite wound strains were similar but not identical to the human B. forsythus ATCC 43037 type strain and the monkey oral strains. The rDNA sequence analysis indicated that the three bite wound isolates had 99.93% homology with each other and 98.9 and 99.22% homology with the human ATCC 43037 and monkey oral strains, respectively. These results suggest that there are host-specific variations within each group.

Animals↗

Microbiota of successful osseointegrated dental implants.

BACKGROUND: The long-term survival of dental implants depends, in part, on control of bacterial infection in the peri-implant region. Periodontal pathogens colonized implants symptomatic through infection, whereas the microbiota of successful implants was similar to that of periodontal health. This study examined the impact on the peri-implant microbiota of crown restorations; implant type; length of time of loading; history of implant or periodontal infections; and whether implants replaced single or multiple teeth. It was of particular interest to evaluate implant colonization by species in a newly described red complex of periodontal pathogens, Porphyromonas gingivalis and Bacteroides forsythus. METHODS: This study sampled 43 partially edentulous subjects with successfully osseointegrated titanium root-form dental implants. Eighty-one (81) non-submerged and 20 submerged asymptomatic implants, 83 crowned, and 36 uncrowned teeth were sampled from peri-implant or subgingival sites. The microbiota of samples was evaluated using whole genomic DNA probes in a checkerboard assay to 23 subgingival species. RESULTS: Implants were colonized principally by oral streptococci, capnocytophagae, Veillonella parvula, Peptostreptococcus micros, and Fusobacterium nucleatum. The periodontal species, P. gingivalis, B. forsythus, Prevotella intermedia, Prevotella nigrescens, and Campylobacter rectus were detected in a few subjects. The microbiota around crowned implants and crowned teeth was similar. Streptococcus oralis, P. intermedia, and Selenomonas noxia were elevated in samples from uncrowned teeth compared to crowned teeth and implants. Microbial complexity increased as loading time increased, but colonization by periodontal pathogens, including red complex species, was higher in subjects with previous periodontal disease. No differences were observed in the microbiota of 1- and 2-stage implants, or between implants supporting single or multiple restorations. CONCLUSIONS: While presence of crowns had only a minor impact on the peri-implant microbiota, microbial changes were observed the longer the implants had been in function and in those patients with a history of periodontal or peri-implant infections. A history of periodontitis had a greater impact on the peri-implant microbiota than implant loading time. The major influence on the peri-implant microbiota was, however, the microbiota on remaining teeth. P. gingivalis and B. forsythus, red complex periodontal pathogens, colonized several implants, although all implants were successfully osseointegrated.

Adult↗

Subgingival temperature and microbiota in initial periodontitis.

The association between subgingival temperature, other clinical characteristics, and the subgingival microbiota was examined in adult subjects with initial periodontitis and differing levels of gingival inflammation. 43 subjects were measured at 6 sites per tooth for pocket depth, attachment level, presence of plaque, gingival redness, bleeding on probing and subgingival temperature at 3-month intervals for 1 year. Subgingival plaque was sampled from 15 initial active periodontitis sites (10 subjects), 121 gingivitis, sites (20 subjects) and 202 healthy sites (13 subjects), and included the 5 hottest and 5 coldest sites in each subject. Plaque samples were analyzed for 13 subgingival species using whole-genomic DNA probes. The major influences on the subgingival microbiota were the clinical status of sites, pocket depth, and the presence of supragingival plaque. No significant association between species and site temperature was observed. Initial active sites were associated with Bacteroides forsythus and Campylobacter rectus, and had a higher mean subgingival temperature and deeper mean pocket depth than inactive sites. A weak association between pocket depth and site temperature was noted. The major influence on subgingival temperature of sites was the anterior to posterior anatomical temperature gradient in the mandible and maxilla.

Adult↗

Microbiota of health, gingivitis, and initial periodontitis.

This study compared the subgingival microbiota in periodontal health, gingivitis and initial periodontitis using predominant culture and a DNA probe, checkerboard hybridization method. 56 healthy adult subjects with minimal periodontal attachment loss were clinically monitored at 3-month intervals for 12 months. More sites demonstrated small increments of attachment loss than attachment gain over the monitoring period. Sites, from 17 subjects, showing > or = 1.5 mm periodontal attachment loss during monitoring were sampled as active lesions for microbial analysis. Twelve subjects demonstrated interproximal lesions, and 5 subjects had attachment loss at buccal sites (recession). Cultural studies identified Bacteroides forsythus, Campylobacter rectus, and Selenomonas noxia as the predominant species associated with active interproximal lesions (9 subjects), whereas Actinomyces naeslundii, and Streptococcus oralis, were the dominant species colonizing buccal active sites. A. naeslundii, Campylobacter gracilis, and B. forsythus (at lower levels than active sites) were the dominant species cultured from gingivitis (10 subjects). Health-associated species (10 subjects) included Streptococcus oralis, A. naeslundii, and Actinomyces gerencseriae. DNA probe data identified higher mean levels of B. forsythus and C. rectus with active (7 subjects) compared to inactive periodontitis sites. Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans were detected infrequently. Cluster analysis of the cultural microbiota grouped 8/9 active interproximal lesions in one subcluster characterized by a mostly gram-negative microbiota, including B. forsythus and C. rectus. The data suggest that B. forsythus C. rectus and S. noxia were major species characterizing sites converting from periodontal health to disease. The differences in location and microbiota of interproximal and buccal active sites suggested that different mechanisms may be involved in increased attachment loss.

Adult↗

Rapid chair-side DNA probe assay of Bacteroides forsythus and Porphyromonas gingivalis.

This study compared a rapid, colorimetric DNA probe assay designed to be performed in a dental office within 40 min, with anaerobic culture and indirect immunofluorescence microscopy (IFM) for detection of Bacteroides forsythus and Porphyromonas gingivalis in subgingival plaque samples. The DNA probe assay used the Periodontal Microbial Identification Test (Saigene Corporation, Bothell, Washington, USA). B. forsythus was detected in 46 (52%), 49 (55%) and 39 (44%) of the samples by DNA probe, culture (at levels > or = 10(5)) and IFM, respectively. P. gingivalis was detected in 24 (27%), 18 (20%) and 29 (33%) of the samples by DNA probe, culture (at levels > or = 10(5)) and IFM, respectively. Results from the DNA probe assay were compared to culture. Culture negative, probe positive samples were re-evaluated by IFM, and IFM positive samples were considered positive in "resolved" data. Using resolved data. DNA probe detection sensitivity and specificity values for B. forsythus were 81% and 91% and for P. gingivalis were 80% and 95%, respectively. DNA probe test results were further compared with culture and IFM. For samples negative by both culture and IFM, probe specificity was 92% in 25 B. forsythus samples and 95% in 57 P. gingivalis samples. For samples positive by both reference methods, probe sensitivity was 82% in 27 B. forsythus samples and 73% in 15 P. gingivalis samples. B. forsythus was detected more frequently by culture compared with IFM; the reverse was observed for P. gingivalis. The rapid DNA probe assay for B. forsythus and P. gingivalis was comparable to cultivable and IF analyses.

Adult↗

Dental implant infections.

Dental implants provide a restorative tool to support crowns, bridge abutments, and removable dentures. Osseointegrated implants are titanium posts that are surgically implanted in alveolar bone. A tight immobile bond (osseointegration) forms between bone and titanium, and prosthetic and restorative fixtures are attached to the implants. Titanium implants differ from natural teeth, which may make them more susceptible to mechanical stress. A small proportion of implants are not successful and may fail due to infection. The microbiota of implants is similar to that of teeth in similar clinical states. Implants that fail because of mechanical stress are colonized by species associated with healthy teeth. Infected implants are colonized by subgingival species, including Porphyromonas gingivalis, Bacteroides forsythus, Fusobacterium nucleatum, Campylobacter gracilis, Streptococcus intermedius, and Peptostreptococcus micros. Different patients may be colonized by different microbial complexes, indicating that optimal treatment should be directed to the specific infection.

Bacterial Infections↗

Clinical, microbiological and immunological profile of healthy, gingivitis and putative active periodontal subjects.

Thirteen periodontally healthy subjects were monitored clinically for 6-12 months. Clinical measurements at 6-weekly intervals included duplicate PD measurements, presence of plaque, redness, and bleeding on probing. Baseline measurements consisted of 2 visits 1 wk apart. Microbial samples were taken from 11 of the subjects who had completed at least 8 months of monitoring. Levels of serum antibodies to 12 periodontal species were determined from 10 subjects. Standard deviations of replicate PD measurements, computed for each subject, ranged from 0.2-0.3 mm over the monitoring period. Plaque and redness increased during monitoring, and showed a weak association with PD change. Baseline and follow-up distributions of PD changes indicated that changes of > 1.5 mm could reasonably be considered to represent active sites. Five subjects demonstrated at least 1 site deepening by 1.5 mm over the period monitored, and these were considered putative active subjects. Sites from 2 subjects showed PD increases in the 6 wk just before sampling, and these were considered to represent active sites. Species associated with putative active subjects included Actinomyces naeslundii, Veillonella parvula, Selenomonas noxia and Prevotella nigrescens. Streptococcus sanguis, S. gordonii and Peptostreptococcus micros were associated with inactive subjects. S. gordonii and S. oralis were associated with health, whereas P. nigrescens was associated with gingivitis. Elevated serum antibodies were detected to A. actinomycetemcomitans in 4 subjects. The predominant microbiota of putative active subjects included some species previously associated with gingivitis, and some species previously associated with progressing periodontitis.

Adult↗

Rapid characterization of periodontal bacterial isolates by using fluorogenic substrate tests.

Eighty-nine species of subgingival bacteria, represented by 121 reference strains and 892 patient isolates, including gram-negative, gram-positive, aerobic, facultatively anaerobic, microaerophilic, and anaerobic species, were characterized with a panel of fluorogenic, 4-methylumbelliferyl-linked substrate tests. Identifications of all patient isolates were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins relative to reference strains. Characteristic profiles of positive fluorogenic reactions differentiated most of the species, including five Porphyromonas species, six pigmenting and five nonpigmenting Prevotella species, Bacteroides forsythus, three Capnocytophaga species, six Actinomyces species, four Propionibacterium species, and eight Streptococcus species. Two mannoside isomers differentiated Actinomyces israelii and Actinomyces gerencseriae. In addition to Porphyromonas gingivalis, B. forsythus, and Capnocytophaga species, Fusobacterium alocis, Actinomyces odontolyticus, Actinomyces meyeri, and Bifidobacterium dentium were all positive for so-called trypsin-like activity. Fusobacterium nucleatum, Eikenella corrodens, Actinobacillus actinomycetemcomitans, and Campylobacter species were nonreactive with the carbohydrate-based substrates tested. Fluorogenic substrate tests provided a sensitive and simple method for biochemical characterization that could presumptively identify to species level most subgingival isolates within 4 h. The method was ideal for rapidly obtaining presumptive identifications of isolates prior to confirming identifications by definitive methods, such as SDS-PAGE.

Bacteria↗

Characterization of hemolytic bacteria in subgingival plaque.

Three-quarters of the patients with periodontal diseases surveyed in this study had one or more distinct types of hemolytic bacteria in their subgingival plaque. Twelve different species of bacteria were identified, belonging to five genera (Actinomyces, Streptococcus, Staphylococcus, Prevotella, and Actinobacillus). Nine hemolytic isolates, consisting of four Prevotella denticola strains, two Actinomyces naeslundii genospecies 2 strains, and one each of P. melaninogenica, Streptococcus constellatus, and A. naeslundii genospecies 1 strains were characterized. Incorporation of pronase into blood agar medium inhibited hemolysis by all of the isolates, suggesting a proteinaceous component for each of their hemolysins. With one exception, hemolysin production appeared to be regulated by the concentration of environmental iron: exogenous hemin was found to inhibit hemolysin production, and the iron scavenging compound, 2,2'- dipyridyl, was found to promote hemolysin production by all of the strains except for the S. constellatus isolate. Genomic libraries of each of the hemolytic plaque isolates were prepared in Escherichia coli using pBR322. Hemolytic clones were isolated on blood agar medium containing ampicillin at frequencies ranging from 1-6.7 x 10(-4). Extensive restriction mapping revealed regions of homology in the case of clones derived from three P. denticola strains isolated from the same subjects. Two of the P. denticola-derived clones were virtually identical throughout the entrety of their > 5 Kb inserts. The clone derived from the third strain showed good homology to the other two within a 1.3 Kb region, but the flanking DNA showed no homology even though all three P. denticola isolates were shown to be clonally related by ribotyping.(ABSTRACT TRUNCATED AT 250 WORDS)

Actinomyces↗

Identification of Selenomonas species by whole-genomic DNA probes, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, biochemical tests and cellular fatty acid analysis.

Nonisotopic, whole-genomic DNA probes, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), biochemical tests in microtiter trays and cellular fatty acid (CFA) analysis were compared for the identification of 5 oral Selenomonas species. DNA probes were prepared by biotin-labeling DNA extracted from the type strains of Selenomonas noxia, Selenomonas flueggei, Selenomonas artemidis, Selenomonas infelix and Selenomonas sputigena. The probes were hybridized with DNA from 21 reference strains, 18 fresh isolates of Selenomonas species, and 21 strains of other oral gram-negative species. Target DNAs were obtained by in situ extraction of colonies blotted onto filter paper. Streptavidin-linked alkaline phosphatase was used to detect homologous reactions of probe and target DNA. Each Selenomonas species DNA probe reacted with reference strains of only that species. All Selenomonas strains that reacted with the DNA probe for a particular species gave similar biochemical test results, SDS-PAGE protein profiles, and CFA profiles to those of the type strain of the corresponding species. All the methods tested were useful for identifying the species, and all yielded similar identifications of the fresh isolates. The DNA probes, however, had the potential for identifying Selenomonas species directly from primary isolation plates or plaque samples.

Bacteroidaceae↗

Identification of oral yeasts by polyacrylamide gel electrophoresis.

Protein profiles of sonicated cells for 9 species of yeast isolated from oral samples were obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and visualized with a silver stain. The profiles obtained on 12% acrylamide gels were distinct, and characteristic for 6 species of Candida, and Torulopsis glabrata, Yarrowia lipolytica, and Saccharomyces cerevisiae. Using this method, 79 fresh isolates of yeasts from saliva samples were identified; 58 as Candida albicans, 9 as Candida parapsilosis, 1 as Candida tropicalis, and 11 as T. glabrata. SDS-PAGE offers a rapid, convenient alternative or adjunct to yeast identification systems based on carbohydrate assimilation tests.

Adult↗

Tetracycline fiber therapy monitored by DNA probe and cultural methods.

Oligonucleotide DNA probe and selective cultural methods were compared in their ability to monitor 6 putative periodontal pathogens in a study evaluating local tetracycline fiber therapy. Subgingival plaque was sampled from 4 sites in each of 20 subjects. Samples were taken before and after therapy from sites assigned to the following test groups: tetracycline (TC) fiber, scaling and root planing, control fiber, and untreated. Each sample was analyzed by both DNA probe and cultural methods. Total anaerobic cultivable counts, Porphyromonas (Bacteroides) gingivalis and Prevotella intermedia (Bacteroides intermedius) were enumerated on nonselective blood agar. Actinobacillus actinomycetemcomitans, Eikenella corrodens, Fusobacterium nucleatum and Wolinella recta were isolated on selective media. TC fiber therapy and scaling reduced total cultivable counts from an initial value of 1 x 10(7) to approximately 2 x 10(5) following therapy. Total counts at untreated sites and at sites with control fibers did not change from baseline. A. actinomycetemcomitans and E. corrodens were detected more frequently by the cultural method; the other monitored species were detected more frequently by DNA probes than by the cultural methods. Agreements between methods were: 77.2% for A. actinomycetemcomitans; 72.2% for P. intermedia; 75.6% for E. corrodens; 39.4% for F. nucleatum; 35.6% for P. gingivalis; and 68.9% for W. recta. Limitations of the selective cultural methods used probably contributed to the discrepancies for P. gingivalis and F. nucleatum. DNA probe and cultural methods indicated comparable levels of suppression of the monitored species following TC fiber therapy and scaling. The microbiota of control fiber and untreated sites did not appear to be significantly altered by either method.

Actinobacillus↗

Detection of high-risk groups and individuals for periodontal diseases: laboratory markers based on the microbiological analysis of subgingival plaque.

Periodontal microbiology is reviewed with regard to the potential of certain characteristics to serve as markers of high risk groups or individuals for periodontal diseases. The generally accepted associations between particular organisms and the various periodontal diseases are discussed. The usefulness of various clinical study designs is reviewed. The ecology of the subgingival plaque microflora is discussed and a number of suggestions for future research are made. We have concluded that there is no monospecific aetiology to any of the various periodontal conditions. Nevertheless, we give particular attention to the role of the black-pigmented bacteroides based upon our belief that they, and Bacteroides gingivalis in particular, are fundamental to our understanding of the biology of periodontal diseases in humans and other animals. Consequently, the contribution of its various virulence factors and their potential as markers of disease susceptibility and activity is addressed.

Bacterial Physiological Phenomena↗

Statistical models for data from periodontal research.

Many factors have been hypothesised either to characterise groups and individuals at risk for periodontal disease or to be markers of periodontal breakdown. In order to identify these as associated either with disease status or progression, a statistical association between the factor and a measure of disease will have to be demonstrated. The statistical modelling of data arising from periodontal research presents special problems. These include the large number of measurements made in each subject, the large magnitude of measurement error compared to the changes in attachment level, the analysis of longitudinal studies, the lack of a measure of instantaneous rate of attachment loss and controversies over the nature of the progression of the disease. We consider statistical methods currently available in the light of these difficulties and identify areas in which further research is necessary.

Analysis of Variance↗