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D H Fine

Publications and source records attributed to D H Fine.

At least 37 records · Page 2Linked to original sources
PubMed1999

Tenacious adhesion of Actinobacillus actinomycetemcomitans strain CU1000 to salivary-coated hydroxyapatite.

Adherence of Actinobacillus actinomycetemcomitans to hard-tissue surfaces was evaluated by comparing a phenotypically stable, well-maintained clinical isolate (strain CU1000) to Streptococcus gordonii G9B, an extensively studied oral-colonizing bacterium. Standard innocula of radiolabelled bacteria were added to saliva-coated hydroxyapatite (SHA) and the ratio of bound to unbound cells counted. Several other clinical isolates as well as laboratory strain Y4 were studied. In other experiments, cell detachment from SHA was compared in static and shaking vessels to calculate controlled desorption of cells over time. A sonic-displacement assay was used to measure avidity of binding to HA and SHA. To better define the attachment properties of CU1000, bacteria were treated with a variety of agents including detergents, salts and enzymes before or after incubation with SHA. Results indicated that CU1000 bound better than G9B (a minimum of 10-fold greater; p < or = 0.05) and did not desorb from SHA, while G9B desorbed to equilibrium in 4 h. Furthermore, Langmuir isotherm calculations indicated that, unlike G9B, CU1000 did not follow second-order adsorption kinetics and thus did not achieve saturation. In addition, of the agents tested only periodate reduced attachment and resulted in detachment of CU1000 from surfaces. These experiments suggest that clinical isolates of A. actinomycetemcomitans possess unique binding properties that promote adsorption to and impede desorption from SHA. The characteristics described for the actinobacillus in this study have been previously underestimated, appear to be mediated by glycoconjugates, and may resemble attachment described for several biofilm-forming, non-oral pathogens.

Adsorption↗
PubMed1999

Direct selection of IS903 transposon insertions by use of a broad-host-range vector: isolation of catalase-deficient mutants of Actinobacillus actinomycetemcomitans.

Transposon mutagenesis in bacteria generally requires efficient delivery of a transposon suicide vector to allow the selection of relatively infrequent transposition events. We have developed an IS903-based transposon mutagenesis system for diverse gram-negative bacteria that is not limited by transfer efficiency. The transposon, IS903phikan, carries a cryptic kan gene, which can be expressed only after successful transposition. This allows the stable introduction of the transposon delivery vector into the host. Generation of insertion mutants is then limited only by the frequency of transposition. IS903phikan was placed on an IncQ plasmid vector with the transposase gene located outside the transposon and expressed from isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible promoters. After transposase induction, IS903phikan insertion mutants were readily selected in Escherichia coli by their resistance to kanamycin. We used IS903phikan to isolate three catalase-deficient mutants of the periodontal pathogen Actinobacillus actinomycetemcomitans from a library of random insertions. The mutants display increased sensitivity to hydrogen peroxide, and all have IS903phikan insertions within an open reading frame whose predicted product is closely related to other bacterial catalases. Nucleotide sequence analysis of the catalase gene (designated katA) and flanking intergenic regions also revealed several occurrences of an 11-bp sequence that is closely related to the core DNA uptake signal sequence for natural transformation of Haemophilus influenzae. Our results demonstrate the utility of the IS903phikan mutagenesis system for the study of A. actinomycetemcomitans. Because IS903phikan is carried on a mobilizable, broad-host-range IncQ plasmid, this system is potentially useful in a variety of bacterial species.

Aggregatibacter actinomycetemcomitans↗
PubMed1998

Codon usage in Actinobacillus actinomycetemcomitans.

The codon usage patterns of 21 genes encompassing 5800 codons from Actinobacillus actinomycetemcomitans were analyzed. A. actinomycetemcomitans genes could be divided into two groups based on their function and G + C content. One group included those genes encoding basic cellular functions. This group displayed an average G + C content of 48%. A second group comprised genes encoding the leukotoxin determinant, an insertion sequence and a plasmid. This group displayed an average G + C content of 36%. These findings suggest that portions of the A. actinomycetemcomitans genome may have been acquired by horizontal gene transfer from one or more distantly related species. We present a table of A. actinomycetemcomitans codon usage. These data may be used to establish standards for computer programs that predict A. actinomycetemcomitans protein coding regions and may be useful in designing degenerate oligonucleotide probes.

Adolescent↗
PubMed1998

Clinical use of antibiotics in dental practice.

Inappropriate use of antibiotics by clinicians leads to development of antibiotic resistance. For the most part, antibiotics are prescribed in dental practice for prophylactic and therapeutic reasons. Prophylactic antibiotics are prescribed to prevent diseases caused by members of the oral flora introduced to distant sites in a host at risk or introduced to a local compromised site in a host at risk. In most cases, prophylaxis is used for prevention of endocarditis. Therapeutic antibiotics are prescribed, in most cases, to treat diseases of hard and soft tissues in the oral cavity after local debridement has failed. Antibiotics used for prophylaxis must: (1) be active against the major pathogens; and (2) achieve a tissue loading dose before the bacteria are introduced. Antibiotics used for therapy are required in cases where the infection is already present and thus the agent must reach the site of infection at a high enough level for a long enough time to produce the desired effect. For an exogenous agent the goal is to eliminate the agent from the site of infection. In the case of an endogenous agent the antibiotic must suppress the organism at the site of infection. Recent evidence underscores the important role of antibiotics in the treatment and prevention of diseases initiated in the oral cavity that have the potential to spread to distant organs in the body.

Antibiotic Prophylaxis↗
PubMed1998

Adjunctive use of a subgingival controlled-release chlorhexidine chip reduces probing depth and improves attachment level compared with scaling and root planing alone.

The present studies evaluated the efficacy of a controlled-release biodegradable chlorhexidine (CHX) (2.5 mg) chip when used as an adjunct to scaling and root planing on reducing probing depth (PD) and improving clinical attachment level (CAL) in adult periodontitis. Two double-blind, randomized, placebo-controlled multi-center clinical trials (5 centers each) were conducted; pooled data are reported from all 10 centers (447 patients). At baseline, following 1 hour of scaling and root planing (SRP) in patients free of supragingival calculus, the chip was placed in target sites with PD 5 to 8 mm which bled on probing. Chip placement was repeated at 3 and/or 6 months if PD remained > or = 5 mm. Study sites in active chip subjects received either CHX chip plus SRP or SRP alone (to maintain study blind). Sites in placebo chip subjects received either placebo chip plus SRP or SRP alone. Examinations were performed at baseline; 7 days; 6 weeks; and 3, 6, and 9 months. At 9 months significant reductions from baseline favoring the chlorhexidine chip compared with both control treatments were observed with respect to PD (chlorhexidine chip plus SRP, 0.95 +/- 0.05 mm; SRP alone, 0.65 +/- 0.05 mm, P < 0.001; placebo chip plus SRP, 0.69 +/- 0.05 mm, P < 0.001) and CAL (chlorhexidine chip plus SRP, 0.75 +/- 0.06 mm; SRP alone, 0.58 +/- 0.06 mm, P < 0.05; placebo chip plus SRP, 0.55 +/- 0.06 mm, P < 0.05). The proportion of patients who evidenced a PD reduction from baseline of 2 mm or more at 9 months was significantly greater in the chlorhexidine chip group (19%) compared with SRP controls (8%) (P < 0.05). Adverse effects were minor and transient toothache, including pain, tenderness, aching, throbbing, soreness, discomfort, or sensitivity was the only adverse effect that was higher in the chlorhexidine group as compared to placebo (P = 0.042). These data demonstrate that the adjunctive use of the chlorhexidine chip results in a significant reduction of PD when compared with both SRP alone or the adjunctive use of a placebo chip. These multi-center randomized control trials suggest that the chlorhexidine chip is a safe and effective adjunctive chemotherapy for the treatment of adult periodontitis.

Adult↗
PubMed1998

Efficacy of a triclosan/NaF dentifrice in the control of plaque and gingivitis and concurrent oral microflora monitoring.

PURPOSE: To compare the effect of a dentifrice containing 0.3% triclosan and 1100 ppm fluoride and a control dentifrice containing 1100 ppm fluoride on plaque, gingiva and the oral microflora in a long-term study simulating clinical usage. MATERIALS AND METHODS: 159 subjects entered the clinical study and 80 were randomly selected to participate in the microbiological evaluation. 71 subjects completed the detailed evaluation of the oral microflora after 6 months use. Plaque was collected at baseline, 3 months, and 6 months, and examined by darkfield microscopy, Gram stain, immunofluorescence, and selective and non-selective media. Changes in antimicrobial susceptibilities were determined for the first 6-month period and for 6 months post-therapy for 68 subjects who completed the entire study. Susceptibilities of whole plaque samples and MIC values for two pre-designated common plaque organisms, A. viscosus and V. parvula were performed. RESULTS: Multivariate ANOVA and non-parametric analyses revealed no statistical differences for any factor tested. No detrimental shifts were found in either; (1) the compositional make up of the normal flora, (2) the periodontopathic or cariogenic flora, or (3) the opportunistic flora in either group of dentifrice users. Both treatments resulted in decreases in Gram positive cocci over time. There was a reduction in spirochetes in the triclosan/fluoride group as compared to the control group. No overgrowth in opportunists, periodontal pathogens, or cariogenic flora was found in either group. No increase in the proportion of the whole plaque flora resistant to triclosan was found nor was an increase in the MIC values of either A. viscosus or V. parvula in either group. Overall, there appeared to be a general decrease in plaque bacteria in both groups over the course of the experiment.

Adolescent↗
PubMed1996

Effects of sublethal exposure to an antiseptic mouthrinse on representative plaque bacteria.

Although the mechanism responsible for the clinical antiplaque efficacy of oral antiseptics is generally considered to be primarily one of bactericidal activity, it has been suggested that oral antiseptics may have additional effects on bacteria exposed to sublethal levels. Studies reported herein, investigated the effects of sublethal levels of an essential oil-containing antiseptic mouthrinse (Listerine Antiseptic, Warner-Lambert Co., Morris Plains, NJ) on selected activities of representative plaque microorganisms using in vitro models. These studies demonstrated that sublethal exposure to the tested oral antiseptic can have significant effects in reducing intergeneric coaggregation, increasing bacterial generation time, and extracting endotoxin from Gram-negative bacteria. These in vitro activities can be correlated with features of plaque formation and pathogenicity seen in vivo; however, additional studies will be necessary to confirm that these mechanisms are, in fact, operative clinically.

Aggregatibacter actinomycetemcomitans↗
PubMed1996

Assessing pre-procedural subgingival irrigation and rinsing with an antiseptic mouthrinse to reduce bacteremia.

In this controlled clinical study, the authors examined the effect of subgingival irrigation and rinsing with an antiseptic mouthrinse before ultrasonic scaling of a quadrant containing inflamed gingivae. The results showed that pre-procedural subgingival irrigation and rinsing can significantly reduce the level of bacteremia associated with ultrasonic scaling. These results support the American Heart Association's recommendation of adjunctive subgingival irrigation prior to invasive procedures in patients at risk of developing bacterial endocarditis.

Adult↗
PubMed1994

Prevalence and distribution of bacteriophage phi Aa DNA in strains of Actinobacillus actinomycetemcomitans.

phi Aa is a bacteriophage that was originally isolated by induction of a lysogenic strain of the oral bacterium Actinobacillus actinomycetemcomitans. Since the discovery of phage phi Aa, additional phages infecting several other strains of A. actinomycetemcomitans have been identified. To determine the prevalence of phi Aa or phi Aa-related temperate phages in this species, a phi Aa-specific DNA probe was prepared to screen for homologous sequences among 42 strains of A. actinomycetemcomitans. Fourteen (33%) of the 42 strains examined contained DNA sequences that hybridized with the phage phi Aa probe. A bacteriophage designated phi Aa33384 was isolated by induction from one of the strains (ATCC 33384) that contained a sequence that hybridized with the phi Aa probe. The phi Aa probe hybridized with the DNA extracted from bacteriophage phi Aa33384. The distribution of the phage phi Aa sequence among A. actinomycetemcomitans serotypes was 5/13 (38%) of the serotype a strains, 0/16 (0%) of the serotype b strains, and 9/13 (69%) of the serotype c strains. The results of this investigation suggest that the target sequence prepared from the phage phi Aa genome is fairly common in the A. actinomycetemcomitans chromosome, and that the sequence is distributed among the A. actinomycetemcomitans serotypes in a seemingly nonrandom manner.

Aggregatibacter actinomycetemcomitans↗
PubMed1994

Microbial identification and antibiotic sensitivity testing, an aid for patients refractory to periodontal therapy. A report of 3 cases.

The importance of microbial surveillance is illustrated in 3 clinical cases. Each case demonstrated a continued lack of response to conventional periodontal treatment. Repeated bouts of periodontal abscess formation and bone loss occurred over a 3- to 4-year period, despite numerous surgeries supplemented with antibiotics. As a result, patients were termed refractory to treatment and extensive microbiological analysis and sensitivity testing was performed. Following institution of the appropriate antibiotic and conservative therapy consisting of several sessions of scaling and root planning, each of these cases demonstrated a dramatic remission of disease progression. No further breakdown has been seen for a minimum of 2 1/2 years. While anecdotal in nature, these cases support the usefulness of microbial identification coupled with antibiotic sensitivity as an adjunct to conventional conservative periodontal therapy.

Adult↗
PubMed1993

Characterization and physical mapping of the genome of bacteriophage phi Aa from Actinobacillus actinomycetemcomitans.

The size, configuration and restriction map of Actinobacillus actinomycetemcomitans bacteriophage phi Aa DNA was determined by means of restriction endonuclease analysis. Digestion of the phi Aa DNA with restriction enzymes Hind III, Eco RI and Sal I produced 6, 5, and 4 fragments, respectively. Based upon the sum of the sizes of the restriction fragments of these enzymes, the DNA was estimated to be 47.2 kilobase pairs in length. A restriction map was constructed using Hind III and Sal I. Incubation with exonuclease Bal 31 for increasing lengths of time resulted in progressive hydrolysis of the DNA, as expected for a linear molecule. No sub-molar fragments or diffuse bands were observed in the agarose gels of the restriction endonuclease digests of the phi Aa DNA. Attempts at ligating the ends of the DNA were consistently unsuccessful. Therefore, we found no evidence for cohesive ends, a circular permutation of the genome or for headful packaging mechanism from a concatameric DNA precursor.

Aggregatibacter actinomycetemcomitans↗
PubMed1993

Identification of Actinobacillus actinomycetemcomitans: polymerase chain reaction amplification of lktA-specific sequences.

Actinobacillus actinomycetemcomitans has been strongly implicated in the etiology of localized juvenile periodontitis. Techniques used in the identification of this periodontal pathogen include cultural, biochemical, immunological and DNA hybridization analysis. In this study, we report the use of polymerase chain reaction (PCR) to amplify unique sequences of A. actinomycetemcomitans. Specific oligonucleotide primers LKT2 and LKT3 were designed to hybridize to the A. actinomycetemcomitans lktA gene, which encodes leukotoxin, a putative A. actinomycetemcomitans virulence factor. The LKT2 and LKT3 primers amplified lktA-specific sequences from all 12 A. actinomycetemcomitans strains tested. In another set of experiments, 13 other bacterial species, most of which are normal residents of the oral cavity, were tested with these primers. These PCR amplifications also contained 2 additional primers, RRN4 and RRN5, which served as positive controls; RRN4 and RRN5 were designed to amplify specific sequences of eubacterial 16S ribosomal DNA (rDNA). PCR amplifications of all bacterial species tested, including A. actinomycetemcomitans, yielded 16S rDNA-specific DNA fragments. Furthermore, each bacterial species tested, with the exception of A. actinomycetemcomitans, failed to amplify lktA sequences. The LKT and RRN primers were used in further PCR experiments to detect A. actinomycetemcomitans directly from gingival fluid samples. The results clearly demonstrate the simplicity, rapidity, specificity and accuracy of the LKT primers in the identification of A. actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans↗
PubMed1993

Conjugal transfer of broad-host-range incompatibility group P and Q plasmids from Escherichia coli to Actinobacillus actinomycetemcomitans.

The first example of conjugal transfer of DNA from Escherichia coli to the periodontal pathogen Actinobacillus actinomycetemcomitans is presented. Derivatives of the incompatibility group P (IncP) plasmid RK2 successfully transferred from an E. coli donor to an A. actinomycetemcomitans recipient. The resulting A. actinomycetemcomitans transconjugants transferred the plasmids back to E. coli recipients. The IncP transfer functions were also used in trans to mobilize the IncQ plasmid pBK1 from E. coli to A. actinomycetemcomitans. The IncP and IncQ plasmids both transferred into A. actinomycetemcomitans at high frequencies (0.3 to 0.5 transconjugants per donor) and showed no gross deletions, insertions, or rearrangements. Determinations of MICs of various antibiotics for the A. actinomycetemcomitans transconjugant strains demonstrated the expression of ampicillin, chloramphenicol, and kanamycin resistance determinants.

Aggregatibacter actinomycetemcomitans↗
PubMed1993

Reduction of viable bacteria in dental aerosols by preprocedural rinsing with an antiseptic mouthrinse.

Two double-blind controlled clinical studies were conducted to determine the efficacy of preprocedural rinsing with Cool Mint Listerine Antiseptic mouthrinse on the level of viable bacteria recovered from dental aerosols when generated immediately after and 40 minutes after rinsing. Eighteen healthy subjects participated in each study. In the first study, following a 24-hour no-oral-hygiene period, subjects received a 10-minute ultrasonic scaling of a randomly chosen half mouth, rinsed with either Cool Mint Listerine or a control, and received an ultrasonic scaling of the remaining half mouth. During each scaling period, aerosolized bacteria were collected on a sterile filter using a modified air-sampling device. The filters were overlaid on culture media, incubated aerobically, and colonies were counted. The second study followed the same basic design except that ultrasonic scaling was done for 5 minutes, and the post-rinsing sampling was performed following a 40-minute simulated dental treatment period. Rinsing with Cool Mint Listerine resulted in a 92.1% reduction in viable bacteria in aerosols generated immediately after rinsing and a 91.3% reduction in aerosols generated 40 minutes after rinsing. These reductions were significantly greater than control (P = 0.0001). These studies suggest that preprocedural rinsing with Cool Mint Listerine Antiseptic may potentially reduce the risk of cross contamination in the dental operatory.

Adult↗
PubMed1992

Efficacy of preprocedural rinsing with an antiseptic in reducing viable bacteria in dental aerosols.

This double-blind, controlled, cross-over, clinical study evaluated the effect of preprocedural rinsing with an antiseptic mouthrinse on the level of recoverable viable bacteria in an aerosol generated during a typical dental procedure. Eighteen subjects participated. Following 24 hours of abstention from all oral hygiene procedures, subjects received a 10-minute ultrasonic scaling of a randomly selected one-half of their mouth which served as the unrinsed control. They were then randomly assigned either antiseptic mouthwash or a control rinse and rinsed with 20 ml for 30 seconds, after which the remaining half mouth (experimental side) was scaled ultrasonically for 10 minutes. During each 10-minute scaling period aerosolized bacteria were collected on a sterile filter using a modified vacuum air-sampling device. Microbes captured on the sterile filter were quantitated by overlaying the filters onto trypticase soy agar, incubating the filters aerobically at 37 degrees C for 24 to 72 hours, and counting the resulting colony forming units (CFU). Preliminary experiments had confirmed that neither the collection method nor residual antiseptic mouthwash in the aerosol adversely affected the number of viable bacteria recovered from the filter. Rinsing with the antiseptic mouthwash produced a 94.1% reduction in recoverable CFUs compared to the non-rinsed control, while the control rinse produced a 33.9% reduction. The difference between the mouthwash and control was statistically significant (P < .001). This study indicates that preprocedural rinsing with an antiseptic mouthwash can significantly reduce the microbial content of aerosols generated during ultrasonic scaling and may have potential in-office use as part of an infection control regimen.

Adult↗