GENETIC APPROACH TO THE STUDY OF EPIDEMIOLOGY AND PATHOGENESIS OF ACTINOBACILLUS ACTINOMYCETEMCOA4ITANS IN LOCALIZED JUVENILE PERIODONTITIS

Actinobacillus acrinomycetemcomirans isolates from periodontal pockets were examined for restriction fragment-length polymorphism using a characterized 4.7-kb DNA probe. A total of 6 patterns of RFLP was found in 133 isolates originating from 12 subjects. No relatedness was found between RFLP types and serotypes. Different periodontal sites within the same subject and different individuals within the same family sometimes showed only one type of A. actinomycetemcomitans RFLP. When members among the same family showed 2 RFLP types, children were always infected with the A. acfinomycefemcomitans strains found in at least one of the parents. These findings support the concept of familial spread of A. actinomycetemcomitans. A. actinomycetemcomitans RFLP type B, corresponding to reference strain JP2, seems to be particularly virulent, as indicated from the presence of RFLP type B in 3 subjects who converted from a healthy periodontal state to localized juvenile periodontitis. RFLP type B was not detected in any of the 21 A. acrinomycetemcomitans-infected patients with adult periodontitis. The RFLP method seems to be useful in determining the epidemiology and possibly the potential virulence of periodontal strains of A. actinomycetemcomitans

Evaluating Two Methods for Fingerprinting Genomes of Actinobacillus Actinomycetemcomitans

The arbitrary primer polymerase chain reaction (AP-PCR) and Southern blot restriction fragment length polymorphism (RFLP) were used to genotype the periodontal pathogen A. actinomycetemcomitans. Total genomic DNA from 73 strains was extracted by conventional methods. Three random-sequence 10-base oligonucleotide primers were chosen for AP-PCR. The amplified DNA products were separated electrophoretically in a 1% agarose gel containing ethidium bromide and the banding patterns were compared among different strains. For RFLP analysis, DNA was digested with EcoRI, separated on a 0.8% agarose gel and transferred to a nylon membrane. The membrane was probed with a previously characterized 5.2 kilobases (kb) DNA fragment cloned from A. actinomycetemcomitans strain Y4. The probe was labeled with digoxigenin, and hybridized fragments were detected with anti-digoxigenin antibody. AP-PCR produced 4–10 DNA bands in the 0.5–5 kb regions and distinguished 9, 13 or 17 genotypes, depending on the specific primer used. Southern blot RFLP analysis revealed 12 hybridization patterns consisting of 1 or 2 DNA fragments (2–23 kb). The addition of the Southern blot analysis to the AP-PCR analysis gave rise to a total of 30 DNA profiles among the 73 A. actinomycetemcomitans study strains. The results indicate that both AP-PCR and Southern blot analysis are useful in clonal analysis of A. actinomycetemcomitans

The identification of bacteria associated with periodontal disease and dental caries by enzymatic methods

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73856/1/j.1399-302X.1986.tb00322.x.pd

Diagnostic potential of chromogenic substrates for rapid detection of bacterial enzymatic activity in health and disease associated periodontal plaques

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65525/1/j.1600-0765.1984.tb01327.x.pd

The role of viruses in oral disease

The focus has traditionally been on bacteria and fungi when discussing microbiological aspects of oral disease. Viruses are probably more involved in diseases associated with the oral cavity than has been previously thought. The role of several viruses in ulceration is well known, but viruses of the herpes family may play a role in periodontitis, and papillomaviruses are probably involved in oral cancer. This review offers a brief introduction to virology before discussing the role of the more relevant viruses in oral disease. As to clinical application, it is concluded that the anti-herpes medication may, in some cases, be relevant in treating periodontitis, while papillomavirus vaccine would be expected to decrease the prevalence of oral cancer

The Role of Spirochetes in Periodontal Disease

The spirochetal accumulation in subgingival plaque appears to be a function of the clinical severity of periodontal disease. It is not known how many different spirochetal species colonize the plaque, but based upon size alone, there are small, intermediate-sized, and large spirochetes. Four species of small spirochetes are cultivable, and of these, T. denticola has been shown to possess proteolytic and keratinolytic enzymes as well as factors or mechanisms which suppress lymphocyte blastogenesis and inhibit fibroblast and polymorphonuclear leukocyte (PMNL) function. All of these attributes could contribute to periodontal tissue insult. Yet independent of these potential virulence mechanisms, the overgrowth of spirochetes can be clinically useful if simply interpreted as indicating the result of tissue damage. In this case, the spirochetes would be indicators of disease and could be easily monitored by microscopic examination of plaque, or possibly by the measurement of benzoyl-DL-arginine-2-naphthylamide (BANA) hydrolytic activity in the plaque.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/68092/2/10.1177_08959374880020021201.pd

Bacteroides fragilis requires the ferrous-iron transporter FeoAB and the CobN-like proteins BtuS1 and BtuS2 for assimilation of iron released from heme

The intestinal commensal and opportunistic anaerobic pathogen Bacteroides fragilis has an essential requirement for both heme and free iron to support growth in extraintestinal infections. In the absence of free iron, B. fragilis can utilize heme as the sole source of iron. However, the mechanisms to remove iron from heme are not completely understood. In this study, we show that the inner membrane ferrous iron transporter ∆feoAB mutant strain is no longer able to grow with heme as the sole source of iron. Genetic complementation with the feoAB gene operon completely restored growth. Our data indicate that iron is removed from heme in the periplasmic space, and the released iron is transported by the FeoAB system. Interestingly, when B. fragilis utilizes iron from heme, it releases heme-derived porphyrins by a dechelatase activity which is upregulated under low iron conditions. This is supported by the findings showing that formation of heme-derived porphyrins in the ∆feoAB mutant and the parent strain increased 30-fold and fivefold (respectively) under low iron conditions compared to iron replete conditions. Moreover, the btuS1 btuS2 doublemutant strain (lacking the predicted periplasmic, membrane anchored CobN-like proteins) also showed growth defect with heme as the sole source of iron, suggesting that BtuS1 and BtuS2 are involved in heme-iron assimilation. Though the dechelatase mechanism remains uncharacterized, assays performed in bacterial crude extracts show that BtuS1 and BtuS2 affect the regulation of the dechelatase-specific activities in an iron-dependent manner. These findings suggest that the mechanism to extract iron from heme in Bacteroides requires a group of proteins, which spans the periplasmic space to make iron available for cellular functions

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

Crevicular fluid pyridinoline cross-linked carboxyterminal telopeptide of type 1 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 m each of 36 subjects. The presence and levels of 40 subgtngivai 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 penodontitts 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. gingivitis, P. intermedia, P. nigrescens and T. dentcola ( p < 0.01). The data indicate that there is a positive relationship between the putative bone resorptive marker ICTP and periodontal pathogens.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/74809/1/j.1600-051X.1998.tb02383.x.pd

The effects on chronic periodontitis of a subgingivally-placed redox agent in a slow release device

Adjunctive chemical agents can reduce the need for meticulous plaque control. The aim of this investigation was to evaluate the periodontal treatment potential of subgingival application of the redox agent methylene blue in a slow release device. This randomized, single-blind, split-mouth study included 18 patients aged 35- 57 years, with chronic adult periodontitis, pocketing of at least 5mm and radiographic evidence of regular bone loss. All experimental sites received subgingival debridement at day 0. Test sites received 32% w/w methylene blue in the slow release device at days 0 and 28. Clinical examination and microbiological sampling were performed at days 0, 7, 28, 56 and 84. Clinical improvements were seen in both groups, but test sites showed consistently greater improvements, some of which were statistically significant (as determined by between-group comparisons utilising SNDs). Significant between-group differences in relation to baseline levels were seen in bleeding index at days 7 and 56, in probable pocket depth at day 56 and for the Perioscan BANA test at day 7. This pilot study thus showed that adjunctive methylene blue in a slow-release device can produce greater clinical and microbiological improvements than subgingival debridement alone.peer-reviewe