PubMed HealthSearch

Biomedical subjects

A D Haffajee

Publications and source records attributed to A D Haffajee.

At least 19 recordsLinked to original sources

Identification of oral Actinomyces species using DNA probes.

Oral Actinomyces comprise a major segment of both the supra- and subgingival microbiota; however, little is known about the distribution of individual species in different sites or clinical conditions. The purpose of the present investigation was to develop DNA probes for suggested species and genotypes of oral Actinomyces. Whole genomic DNA probes to 12 human oral species and/or serotypes were labeled with digoxigenin and used to seek cross-reactions among the taxa using the checkerboard DNA-DNA hybridization assay. The Actinomyces formed three distinct groups: 1) Actinomyces georgiae, Actinomyces meyeri and Actinomyces odontolyticus serotypes I and II; 2) Actinomyces viscosus and Actinomyces naeslundii serotypes I, II, III and WVA 963; and 3) Actinomyces gerencseriae and Actinomyces israelii. Cross-reactions among taxa were detected and minimized by increasing the temperature of the post-hybridization high-stringency wash to 80 degrees C. Despite the elevation in high stringency wash temperature, cross-reactions among strains of the A. naeslundii/A. viscosus group persisted. Probes for two of the three currently recognized genospecies in this group were prepared by removing the DNA in common between cross-reacting species using subtraction hybridization and polymerase chain reaction. Nine species and genospecies could be clearly separated by a combination of whole genomic and subtraction hybridization probes and by increasing the high-stringency wash temperature. A total of 195 fresh isolates of Actinomyces were grouped in a blind study using DNA probes and separately by SDS-PAGE protein profiles. Concordance between the two methods was 97.3%. The probes and hybridization conditions were tested for their ability to detect the Actinomyces species and genospecies in samples of supragingival and subgingival plaque from periodontitis subjects using checkerboard DNA-DNA hybridization. The probes detected the species in samples of supragingival and subgingival plaque. We concluded that whole genomic and subtraction hybridization DNA probes facilitate the detection and enumeration of species and genospecies of Actinomyces in plaque samples.

Actinomyces

Discrimination of refractory periodontitis subjects using clinical and laboratory parameters alone and in combination.

The purpose of the present investigation was to use baseline clinical and laboratory parameters to distinguish subjects refractory to conventional periodontal therapy. Baseline clinical, microbial and host parameters were compared in 61 successfully-treated and 27 refractory subjects. Refractory subjects showed mean full-mouth attachment level (AL) loss and/or >3 sites with new AL loss >2.5 mm within 1 year after both scaling and root planing and surgery with systemic tetracycline. Successfully-treated subjects showed mean AL gain and no sites with new AL loss >2.5 mm after either regimen. Gingival redness, bleeding on probing, suppuration, supragingival plaque accumulation, pocket depth and AL were measured at 6 sites per tooth in each subject. The levels of 40 subgingival taxa were determined in subgingival plaque samples from up to 28 sites in each subject using checkerboard DNA-DNA hybridization. Serum antibody (Ab) to 85 subgingival species was determined using checkerboard immunoblotting. Levels of serum IgG2 and Gm23 allotype were measured using radial immunodiffusion; FcgammaRIIa and FcgammaRIIIb receptor haplotypes were determined using PCR and allele specific oligonucleotide probes. Odds ratios of a subject being refractory were determined by comparing measured parameters in the 2 subject groups using univariate and multivariate techniques. 17 of 151 clinical, microbial and immunological variables were significant using chi2 analysis after adjusting for multiple comparisons. For example, the odds ratios of a subject being refractory were 12.2, 5.4 and 6.9 if the subject had Ab >50 microg/ml to >9 species; S. constellatus counts >2.4% of the total DNA probe count or >2.1% of sites with AL >6 mm. The 17 significant predictor variables were used in logistic regression and discriminant analyses. Similar variables were selected using both analyses including the number of serum Ab to subgingival species >50 microg/ml, % S. constellatus in plaque samples and % sites with attachment loss >6 mm. In the logistic regression analysis model, the odds ratios associated with >9 species exhibiting >Ab 50 microg/ml, >2.1% of sites with AL >6 mm and >2.4% S. constellatus in plaque were 8.7, 6.8 and 2.4, respectively, after adjusting for other variables in the model. Discriminant analysis using these variables provided sensitivity, specificity, positive and negative predictive values of 0.66, 0.92, 0.80 and 0.85 respectively. Refractory periodontitis subjects could be distinguished using a subset of clinical, microbiological and immunological parameters.

Adult

Relationship between C-telopeptide pyridinoline cross-links (ICTP) and putative periodontal pathogens in periodontitis.

Crevicular fluid pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP) is predictive for future alveolar bone loss in experimental periodontitis in dogs. The present study sought to relate ICTP to a panel of subgingival species in subjects exhibiting various clinical presentations such as health (n=7), gingivitis (n=8) and periodontitis (n=21). 28 subgingival plaque and GCF samples were taken from mesiobuccal sites in each of 36 subjects. The presence and levels of 40 subgingival taxa were determined in plaque samples using whole genomic DNA probes and checkerboard DNA-DNA hybridization. GCF ICTP levels were quantified using radioimmunoassay (RIA). Clinical assessments made at the same sites included: BOP, gingival redness, plaque, pocket depth, and attachment level. Differences among ICTP levels in the 3 subject groups were sought using the Kruskal-Wallis test. Relationships between ICTP levels and clinical parameters as well as subgingival species were determined by regression analysis. The results demonstrated significant differences among disease categories for GCF ICTP levels for healthy (1.1+0.6 pg/site (mean+/-SEM)) gingivitis (14.8+/-6.6 pg/site) and periodontitis subjects (30.3+5.7 pg/site) (p= 0.0017). ICTP levels related modestly to several clinical parameters. Regression analysis indicated that ICTP levels correlated strongly with mean subject levels of several periodontal pathogens including B. forsythus, P. gingivalis, P. intermedia, P. nigrescens and T. denticola (p<0.01). The data indicate that there is a positive relationship between the putative bone resorptive marker ICTP and periodontal pathogens.

Adult

Microbial complexes in subgingival plaque.

It has been recognized for some time that bacterial species exist in complexes in subgingival plaque. The purpose of the present investigation was to attempt to define such communities using data from large numbers of plaque samples and different clustering and ordination techniques. Subgingival plaque samples were taken from the mesial aspect of each tooth in 185 subjects (mean age 51 +/- 16 years) with (n = 160) or without (n = 25) periodontitis. The presence and levels of 40 subgingival taxa were determined in 13,261 plaque samples using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments were made at 6 sites per tooth at each visit. Similarities between pairs of species were computed using phi coefficients and species clustered using an averaged unweighted linkage sort. Community ordination was performed using principal components analysis and correspondence analysis. 5 major complexes were consistently observed using any of the analytical methods. One complex consisted of the tightly related group: Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola. The 2nd complex consisted of a tightly related core group including members of the Fusobacterium nucleatum/periodonticum subspecies, Prevotella intermedia, Prevotella nigrescens and Peptostreptococcus micros. Species associated with this group included: Eubacterium nodatum, Campylobacter rectus, Campylobacter showae, Streptococcus constellatus and Campylobacter gracilis. The 3rd complex consisted of Streptococcus sanguis, S. oralis, S. mitis, S. gordonii and S. intermedius. The 4th complex was comprised of 3 Capnocytophaga species, Campylobacter concisus, Eikenella corrodens and Actinobacillus actinomycetemcomitans serotype a. The 5th complex consisted of Veillonella parvula and Actinomyces odontolyticus. A. actinomycetemcomitans serotype b, Selenomonas noxia and Actinomyces naeslundii genospecies 2 (A. viscosus) were outliers with little relation to each other and the 5 major complexes. The 1st complex related strikingly to clinical measures of periodontal disease particularly pocket depth and bleeding on probing.

Adult

Clinical and microbiological features of refractory periodontitis subjects.

The purpose of this investigation was to compare the clinical parameters and the site prevalence and levels of 40 subgingival species in successfully treated and refractory periodontitis subjects. 94 subjects received scaling and root planing and if needed, periodontal surgery and systemically administered tetracycline. 28 refractory subjects showed mean full mouth attachment loss and/or > 3 sites showing attachment loss > 2.5 mm within 1 year post-therapy. 66 successfully treated subjects showed mean attachment level gain and no sites with attachment loss > 2.5 mm. Baseline subgingival plaque samples were taken from the mesial aspect of each tooth and the presence and levels of 40 subgingival taxa were determined using whole genomic DNA probes and checkerboard DNA-DNA hybridization. The mean levels and % of sites colonized by each species (prevalence) was computed for each subject and differences between groups sought using the Mann-Whitney test. Most of the 40 species tested, including Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola and Bacteroides forsythus, were equally or less prevalent in the refractory group. Prevotella nigrescens was significantly more prevalent in successfully treated subjects, while refractory subjects harbored a larger proportion of Streptococcus species, particularly Streptococcus constellatus. The odds of a subject being refractory was 8.6 (p < 0.001) if S. constellatus constituted > or = 3.5% of the total DNA probe count. Since few microbiological differences existed between treatment outcome groups using DNA probes to known species, the predominant cultivable microbiota of 33 subgingival samples from 14 refractory subjects was examined. 85% of the 1649 isolates were identified using probes to 69 recognized subgingival species. The remaining unidentified strains were classified by analyzing 16S rRNA gene sequences. Many sequenced isolates were of taxa not considered a common part of the oral microbiota such as Acinetobacter baumanni, Gemella haemolysans, Enterococcus faecalis, Staphylococcus warneri, Pseudomonas aeruginosa and novel species in the genera Bartonella, Ralstonia, Neisseria, Eubacterium, Rothia, Gordona, Gemella, Corynebacterium, Leptotrichia, and Actinomyces. Refractory subjects constituted a heterogeneous group based on their subgingival microbiota. As a group, they did not harbor more of the "classic" periodontopathogens, although elevated proportions of S. constellatus were found.

Adult

Subgingival microbiota in healthy, well-maintained elder and periodontitis subjects.

This investigation compared the site prevalence of 40 subgingival species in 30 periodontally healthy (mean age 36+/-9 years), 35 elders with a well-maintained periodontium (mean age 77+/-5) and 138 adult periodontitis subjects (mean age 46+/-11). Subgingival plaque samples were taken from the mesial aspect of each tooth (up to 28 samples) in the 203 subjects at baseline. The presence and levels of 40 subgingival taxa were determined in 5003 plaque samples using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments including dichotomous measures of gingival redness, bleeding on probing, plaque accumulation and suppuration, as well as duplicate measures of pocket depth and attachment level, were made at 6 sites per tooth. The % of sites colonized by each species (prevalence) was computed for each subject. Differences in prevalence and levels among groups were sought using the Kruskal-Wallis test. Commonly detected species, such as Actinomyces naeslundii genospecies 2, Streptococcus sanguis and Streptococcus oralis did not differ significantly among subject groups. After adjusting for multiple comparisons, 4 species were significantly elevated and at greater prevalence in the periodontitis group. Mean % of sites (+/-SEM) colonized by Bacteroides forsythus was 10+/-3, 12+/-2 and 40+/-2 (p<0.001) for healthy, elder and periodontitis groups respectively. The odds ratio was 14.4:1 that a subject had periodontitis when B. forsythus was detected at > or = 5% of sampled sites. Mean prevalence for Porphyromonas gingivalis in healthy, elder and periodontitis subjects was 4+/-2, 5+/-2 and 23+/-2 respectively (p<0.001); for Treponema denticola 12+/-4, 10+/-3 and 30+/-2 (p<0.001) and for Selenomonas noxia 6+/-2, 7+/-2 and 19+/-2 (p<0.01). Similar differences among subject groups were observed when only sites with PD 0-4 mm were analyzed. The data suggest an etiologic role for B. forsythus, P. gingivalis, T. denticola and S. noxia in adult periodontitis.

Actinomyces

Serum IgG2 level, Gm(23) allotype and FcgammaRIIa and FcgammaRIIIb receptors in refractory periodontal disease.

The purpose of this investigation was to compare the levels of serum IgG2, the frequency of detection of Gm(23)-negative allotype and frequency of detection of FcgammaRIIa and FcgammaRIIIb receptor haplotypes in 32 refractory, 54 successfully treated and 27 periodontally healthy individuals. Refractory subjects showed mean full mouth attachment loss and/or >3 sites with attachment loss >2.5 mm within 1 year after both scaling and root planing, and surgery plus systemically administered tetracycline. Successfully treated subjects showed mean attachment level gain and no sites with attachment loss >2.5 mm 1 year post-therapy. Periodontally healthy subjects exhibited no pocket depth or attachment level >3 mm, and no evidence of progressing disease during 1 year of monitoring. Blood was obtained from each subject at baseline. Serum IgG2 and Gm(23) allotype were determined using radial immunodiffusion. DNA was extracted from whole blood and the FcgammaR genotypes determined using PCR and allele specific oligonucleotide probes. Significance of differences among clinical groups were sought using the Kruskal-Wallis or chi-square tests. Associations between 2 or more variables were tested using regression analysis. Refractory subjects exhibited higher mean attachment loss and pocket depth than successfully treated or periodontally healthy subjects. Smoking status did not differ significantly among groups. No significant differences in serum IgG2 levels and frequency of detection of Gm(23)-negative allotype were observed among the clinical groups. Serum IgG2 level was positively associated with the number of serum antibody responses to subgingival species (r=0.51, p<0.001). Subjects with the Gm(23)-negative allotype exhibited lower mean levels of serum IgG2 (3.06+/-0.3 versus 3.9+/-0.2, p<0.01) and mean number of serum antibodies to subgingival species (17.7+/-1.7 versus 23.3+/-1.4, p<0.05) than allotype positive individuals. No significant differences in FcgammaR haplotype distribution were observed among the 3 clinical groups. Associations of serum IgG2 level, Gm(23) allotype, FcgammaRIIa and FcgammaRIIIb receptor haplotypes and smoking status were weakly related or not related to clinical status. This lack of relationship may have been due to a reality of no relationship, or the inadvertent pooling of subjects where these factors were of primary importance with subjects in whom these factors played a less important role.

Adult

Serum antibodies reacting with subgingival species in refractory periodontitis subjects.

The purpose of this investigation was to compare the levels of serum IgG antibody to 85 subgingival species in 32 refractory periodontitis, 56 successfully treated, and 33 periodontally healthy subjects. Refractory subjects showed mean full mouth attachment loss and/or >3 sites showing attachment loss >2.5 mm within 1 year after 2 treatment modalities, scaling and root planing and surgery plus systemically administered tetracycline. Successfully-treated subjects showed mean attachment level gain and no sites with attachment loss >2.5 mm, 1 year post-therapy. Periodontally healthy subjects exhibited no pocket or attachment level >3 mm, and no evidence of progressing attachment loss during 1 year of monitoring. Baseline serum was obtained from each subject and tested against 85 subgingival species, including reference strains and strains isolated from refractory subjects, using checkerboard immunoblotting. Significance of differences in levels of serum antibody among groups were sought using the Kruskal-Wallis test. Refractory subjects constituted a heterogeneous group based on their serum antibody response to subgingival species. Some individuals had antibody reactions to many subgingival species, while other subjects showed fewer or low numbers of responses. On average, refractory subjects exhibited higher numbers and levels of serum antibody reactions to a wide range of subgingival species than successfully treated or periodontally healthy subjects. Differences in serum antibody among clinical groups were more striking at higher threshold levels of antibody (>50 microg/ml and > 100 microg/ml). The data showed that a subject was 10.1 x more likely to be refractory if the subject exhibited antibody reactions with >9 subgingival species at >50 microg/ml (p<0.001, after adjusting for multiple comparisons). Serum antibody to a subset of the test species differed among the clinical groups. Porphyromonas gingivalis, Bacteroidesforsythus, and some strains isolated from refractory subjects (a novel Neisseria sp., Enterococcus faecalis, Prevotella loescheii and Prevotella oulora) elicited high serum antibody in the successfully treated and refractory subjects. High levels of serum antibody to a Microbacterium lacticum-like organism, Streptococcus oralis, Streptococcus constellatus, Actinobacillus actinonmycetemcomitans serotype c and Haemophilus aphrophilus significantly increased the likelihood of a subject being refractory to conventional periodontal therapy.

Adult

Detection of identical ribotypes of Porphyromonas gingivalis in patients residing in the United States, Sudan, Romania and Norway.

Porphyromonas gingivalis has been isolated from periodontitis lesions in subjects from many geographical locations. The purpose of this investigation was to determine whether similar ribotypes of P. gingivalis could be detected among strains isolated in different countries. A total of 198 isolates of P. gingivalis were obtained from 52 periodontitis patients in Boston (130 isolates), Bergen, Norway (17 isolates), Khartoum, Sudan (26 isolates), and Bucharest, Romania (25 isolates). DNA was isolated from each strain, cut separately by the restriction endonucleases KpnI and PstI. The resulting preparations were subjected to electrophoresis in a 0.8% agarose gel using a Tris-acetate EDTA buffer. Uncut lambda and a 1000-bp fragment of 16S rRNA were included as internal standards in each lane. In addition, a HindIII digest of lambda was present in a separate lane in each run. The DNA fragments were transferred to a nylon membrane by downward capillary transfer. 16S rRNA bands were detected using a 1000-kb digoxigenin-labelled probe generated by a polymerase chain reaction. At the same time, a digoxigenin-labelled probe to lambda was employed to detect the internal and molecular weight standards. The bands were detected using antibody to digoxigenin conjugated to alkaline phosphatase and chemiluminescence. The positions of the bands relative to the internal standards were determined and normalized to correct for run-to-run variations, and the molecular weight of each band was determined by comparison with standards within each gel. The resulting data for the 2 enzymes were combined and subjected to cluster analysis using an average unweighted linkage sort. In some instances, isolates that appeared to be of identical ribotype using one endonuclease gave different ribotypes using the other. Strains of P. gingivalis within a subject were usually identical, except for 3 patients who harbored 2 different ribotypes/individual. All subsequent analyses employed a single ribotype strain for each subject. A total of 32 ribotypes were observed for isolates from distant countries. A total of 11.5% of the patients had isolates exhibiting the same ribotype: ribotype 7a. Identical ribotypes of P. gingivalis can be recovered from subgingival plaque samples of periodontitis patients in different countries.

Bacterial Typing Techniques

Clinical and microbiological features of subjects with adult periodontitis who responded poorly to scaling and root planing.

In a previous report, it was shown that scaling and root planing (SRP) decreased mean pocket depth and attachment level in subjects with adult periodontitis, as well as the levels and prevalence of Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola. However, a subset of subjects in that study exhibited mean loss of attachment following SRP. The purpose of the present investigation was to seek clinical and microbiological differences between subjects who responded well or poorly to SRP. 57 subjects with adult periodontitis were treated by full-mouth SRP under local anaesthetic. Clinical assessments of plaque, redness, suppuration, BOP, pocket depth and attachment level were made at 6 sites per tooth prior to and 3 months post-SRP. Attachment level measurements were repeated at each visit and differences in means between visits used to assess change. 18 subjects showed mean attachment loss 3 months post-SRP (poor response group), while 39 showed mean attachment level gain (good response group). The prevalence and levels of 40 subgingival taxa in subgingival plaque samples from the mesiobuccal site of each tooth (maximum 28 sites) in each subject prior to and 3 months post-SRP were assessed using checker-board DNA-DNA hybridization. The prevalence of each species was computed for each subject and averaged across subjects in the 2 treatment-response groups at each visit. Differences between groups were sought using the Mann-Whitney test. There were no statistically significant differences between the 2 response groups in any clinical parameter prior to therapy. Subjects in the good response group showed more attachment level gain at sites with baseline pocket depths of < 4 mm, 4-6 and > 6 mm than poor response subjects. Of 40 species evaluated, A. naeslundii genospecies 2 (A. viscosus), T. denticola, C. gracilis and C. rectus were significantly higher and more prevalent pre-therapy in the good response subjects. Mean attachment level change post SRP could be predicted using multiple linear regression with A. naeslundii genospecies 2 (A. viscosus) and T. denticola as the predictor variables (r2 = 0.373, p < 0.00001). Sites that gained > or = 2 mm of attachment post therapy showed a significant decrease in the counts of P. gingivalis (7.5 +/- 3.5 to 0.2 +/- 0.2 x 10(5)), T. denticola (8.2 +/- 3.5 to 1.8 +/- 1.1 x 10(5)) and B. forsythus (11.1 +/- 5.7 to 0.3 +/- 0.2 x 10(5)). The data of the present investigation indicate that SRP is most effective in subjects and sites with high levels of the subgingival species that this therapy affects.

Actinomyces viscosus

The effect of SRP on the clinical and microbiological parameters of periodontal diseases.

The purpose of the present investigation was to examine the effect of SRP on clinical and microbiological parameters in 57 subjects with adult periodontitis (mean age 47 +/- 11 years). Subjects were monitored clinically and microbiologically prior to and 3, 6 and 9 months after full-mouth SRP under local anaesthesia. Clinical assessments of plaque, redness, suppuration, BOP, pocket depth and attachment level were made at 6 sites per tooth. The means of duplicate attachment level measurements taken at each visit were used to assess change between visits. Clinical data were averaged within each subject and then averaged across subjects for each visit. Subgingival plaque samples were taken from the mesial aspect of each tooth and the presence and levels of 40 subgingival taxa were determined using whole genomic DNA probes and checkerboard DNA-DNA hybridization. The mean levels and % of sites colonized by each species (prevalence) was computed for each subject at each visit. Differences in clinical and microbiological parameters before and after SRP were sought using the Wilcoxon signed ranks test or the Quade test for more than 2 visits. Overall, there was a mean gain in attachment level of 0.11 +/- 0.23 mm (range -0.53 to 0.64 mm) 3 months post-therapy. There was a significant decrease in the % of sites exhibiting gingival redness (68 to 57%) and BOP (58 to 52%) as well as a mean (+/-SEM) pocket depth (3.3 +/- 0.06 to 3.1 +/- 0.05 mm). Sites with pre-therapy pocket depths of < 4 mm showed a non-significant increase in pocket depth and attachment level, 4.6 mm pockets showed a significant decrease in pocket depth and a non-significant gain in attachment post-therapy, while > 6 mm pockets showed a significant decrease in pocket depth and attachment level measurements post-therapy. Significant clinical improvements were seen in subjects who had never smoked or were past smokers but not in current smokers. Mean prevalences and levels of P. gingivalis, T. denticola and B. forsythus were significantly reduced after SRP, while A. viscosus showed a significant increase in mean levels. The mean decrease in prevalence of P. gingivalis was similar at all pocket depth categories, while B. forsythus decreased more at shallow and intermediate pockets and A. viscosus increased most at deep sites. P. gingivalis. B. forsythus and T. denticola were equally prevalent among current, past and never smokers pre-therapy, decreased significantly post-SRP in never and past smokers but increased in current smokers. Clinical improvement post-SRP was accompanied by a modest change in the subgingival microbiota, primarily a reduction in P. gingivalis, B. forsythus and T. denticola, suggesting potential targets for therapy and indicating that radical alterations in the subgingival microbiota may not be necessary or desirable in many patients.

Adult

The nature of periodontal diseases.

Periodontal diseases are infections. They have a number of properties in common with infections in other parts of the body, but have unique features resulting from the passage of the tooth through the soft tissue integument into the oral cavity. The tooth provides a solid, non-shedding surface for the colonization of potentially pathogenic bacterial species as well as a wide range of host-compatible species. Periodontal pathogens frequently colonize the periodontal area for prolonged periods of time prior to disease initiation. Disease is caused by a finite set of bacterial species leading to the development of multiple periodontal diseases. Disease occurs at individual periodontal sites and leaves an historical record of the damage to the periodontium in the form of periodontal attachment or bone loss. This feature provides difficulty in investigation since it is often unclear whether the infectious process is presently occurring or is a reflection of past destruction. Control of periodontal diseases is most effectively performed by eliminating or suppressing the organisms that cause them and establishing a host compatible microbiota. When tissue damage has been extensive, regenerative or replacement procedures should be instituted after the infection has been controlled.

Bacterial Infections

Response to periodontal therapy in patients with high or low levels of P. gingivalis, P. intermedia, P. nigrescens and B. forsythus.

In a previous study, subjects receiving either adjunctive tetracycline or Augmentin showed, on average, more attachment level gain 10 months post-therapy than subjects receiving either Ibuprofen or a placebo, although some subjects in each treatment group showed loss of attachment post-therapy. Since differences in treatment response might have been due to differences in the subgingival microbiota, the response to different therapies in subjects with different pre-therapy subgingival microbiotas was evaluated. 29 subjects exhibiting loss of attachment > 2.5 mm at 1 or more sites during longitudinal monitoring were treated by modified Widman flap surgery at deep sites, subgingival scaling at all other sites and were randomly assigned one of the following agents: Augmentin, tetracycline, ibuprofen or a placebo. Treatment was completed within 30 days, during which time the subject took the assigned agent. Subgingival plaque samples were taken from the mesial surface of each tooth at each visit and evaluated for their content of 14 subgingival species including P. gingivalis, P. nigrescens, P. intermedia and B. forsythus using DNA probes. 18 subjects with mean counts > 10(5) of 2 or more of these 4 species comprised the high test species group; 11 subjects with mean counts > 10(5) of 0 or 1 of the species, the low test species group. Because this was a post-hoc analysis, the number of subjects in some of the treatment/test species groups was small. However, the 8 high test species subjects who received tetracycline showed the most attachment level gain (0.83 +/- 0.20 mm), while the 3 tetracycline-treated, low test species subjects showed minimal gain (0.05 +/- 0.28 mm) 10 months post-therapy. Low test species subjects receiving Augmentin (n = 2) showed a mean gain in attachment of 0.67 (+/- 0.59) mm. The mean % of sites showing either attachment gain or loss > or = 2 mm was computed for each treatment/test species group. High test species subjects receiving tetracycline exhibited the best ratio of gaining to losing sites (16.2), followed by low test species subjects receiving Augmentin (14.1). Periodontal pockets < 7 mm pre-therapy in low test species subjects treated with Augmentin and high test species subjects treated with tetracycline showed attachment gain more frequently than attachment loss. The greatest proportion of gaining sites was seen at pockets > 6 mm, particularly in subjects receiving adjunctive tetracycline. Overall, the data indicated that a gain in mean attachment level post-therapy was significantly associated (p < 0.001) with an increase in C. ochracea accompanied by a decrease in B. forsythus, P. gingivalis, P. intermedia and P. nigrescens. The 4 test species were decreased more in subjects receiving tetracycline. In contrast, Augmentin appeared to be effective in decreasing the % sites colonized by A. actinomycetemcomitans and in increasing the proportion of sites colonized by C. ochracea. Knowledge of the baseline microbiota should improve the choice of an appropriate adjunctive antibiotic for periodontal therapy.

Adolescent

Patterns of antibody response in subjects with periodontitis.

Periodontal diseases comprise a heterogeneous group of infections that are difficult to distinguish on a clinical basis alone. The purpose of the present investigation was to group periodontitis subjects according to their elevated serum antibody levels to specific subgingival species. A total of 119 subjects (19-70 years) with evidence of prior periodontal destruction were monitored at 2-month intervals (maximum 8 visits), prior to therapy, using clinical parameters measured at 6 sites per tooth. The probing attachment level was measured twice at each visit, and an increase of > 2.5mm at a site was used to define subjects with progressing disease. Serum samples were obtained from each subject at each visit and the level of antibody determined by enzyme-linked immunosorbent assay to 12 subgingival species. Subgingival plaque samples were taken from the mesial aspect of all teeth in each subject at each visit, and the levels of 14 different subgingival species were determined using a colony-lift method and DNA probes. Subjects were grouped by cluster analysis of their elevated antibody levels using a simple matching coefficient. Ninety-two subjects fell into 9 clusters with 100% similarity; 29 subjects in one cluster group exhibited elevated antibody to none of the test species. Seven subjects in a second cluster group showed elevated antibody to Bacteroides forsythus. Subjects in the other 7 clusters showed elevated antibody to Actinobacillus actinomycetemcomitans serotype a only or in combination with B. forsythus, A. actinomycetemcomitans serotype b, Prevotella intermedia or Porphyromonas gingivalis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Clinical and microbiological changes associated with the use of 4 adjunctive systemically administered agents in the treatment of periodontal infections.

The purpose of the present investigation was to assess the effects of periodontal surgery and 4 systemically administered agents, Augmentin, tetracycline, ibuprofen or a placebo on clinical and microbiological parameters of periodontal disease. 98 subjects were monitored at 2-month intervals at 6 sites per tooth for clinical parameters. Subgingival plaque samples were taken from the mesial surface of each tooth at each visit and evaluated for their content of 14 subgingival species using DNA probes and a colony lift method. 40 subjects who exhibited loss of attachment > 2.5 mm at 1 or more sites during longitudinal monitoring were treated using modified Widman flap surgery at sites with probing pocket depth > 4 mm, subgingival scaling at all other sites and were randomly assigned 1 of the 4 agents. Treatment was completed within 30 days during which time the subject took the assigned agent. Overall, subjects exhibited a mean attachment level "gain" of 0.34 +/- 0.10 mm (SEM) and a mean pocket depth reduction of 0.62 +/- 0.09 mm 10 +/- 4 months post-therapy. However, certain subjects in each treatment group showed a poor response. Subjects receiving antibiotics exhibited significantly more attachment level "gain" (0.57 +/- 0.15 mm, SEM) than subjects receiving either ibuprofen or a placebo (0.02 +/- 0.10). The differences between Augmentin and tetracycline groups were not significant, nor were the differences between ibuprofen and placebo. 10 months post-therapy, there was a reduction in the number of sites colonized in any subject group by detectable levels (10(3)) of P. gingivalis. Species showing similar reductions were B. forsythus, P. intermedia and P. micros. Subjects receiving systemically administered antibiotics had a significant increase in the proportion of sites colonized by C. ochracea coupled with a greater decrease in the number of sites colonized by P. gingivalis, B. forsythus, P. intermedia and P. micros post-therapy than subjects not receiving antibiotics. The results of this investigation indicate that adjunctive systemic antibiotics increase periodontal attachment "gain" and decrease the levels of some suspected periodontal pathogens in subjects with evidence of current disease progression.

Adolescent

Factors associated with different responses to periodontal therapy.

In a study of the efficacy of modified Widman flap surgery and scaling and root planning accompanied by 1 of 4 systemic adjunctive agents, Augmentin, tetracycline, ibuprofen or placebo, it was observed that subjects differed in their response to therapy. The difference was only partially accounted for by the adjunctive agent employed. The purpose of the present investigation was to examine clinical and microbiological features in subjects who showed different levels of attachment change post-therapy. 40 subjects were subset into 3 groups based on mean attachment level change post-therapy. 10 poor response subjects showed mean attachment loss; 19 moderate response subjects showed mean attachment gain between 0.02-0.5 mm and 11 good response subjects showed a mean gain of attachment > 0.5 mm. Clinical parameters were measured at 6 sites per tooth both pre- and post-therapy. Microbiological samples were taken from the mesial aspect of each tooth and evaluated individually for their content of 14 subgingival taxa using a colony lift method and DNA probes. % of sites colonized by each species was computed for each subject both pre- and post-therapy. Significant differences were observed among treatment response groups for mean probing pocket depth, attachment level and % of sites with plaque pre-therapy. The poor response subjects had the lowest mean probing pocket depth and attachment level, but the highest plaque levels. Post-therapy, the poor response group exhibited the greatest degree of gingival inflammation as assessed by gingival redness and bleeding on probing.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent