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Human papillomavirus in oral premalignant lesions.

The aetiology of oral premalignant lesions is generally accepted to be multifactorial. Tobacco and alcohol are established as important cofactors in malignant development in the oral cavity, but in addition microorganisms, such as human papillomavirus (HPV), have gained much interest over the past decade. For many years, HPV has been accepted as an important cofactor in the development of cervical cancer, originating from a mucous membrane with similarities to the oral mucosa. 49 patients with oral premalignant lesions and 20 control patients with normal oral mucosa and no history of HPV infection were examined for the presence of HPV by immune histochemical staining using the peroxidase anti-peroxidase technique (PAP), DNA-DNA in situ hybridisation (ISH), and polymerase chain reaction (PCR) analysed by Southern blot hybridisation with an HPV 16 specific probe. The investigations revealed that HPV was found in 62.5% of the verrucous leucoplakias, 50.0% of the erythroplakias, 45.5% of the homogeneous leucoplakias, 33.3% of erythroleucoplakias and in 12.5% of the nodular leucoplakias. An overall HPV detection rate in the examined premalignant lesions was 40.8% and no patients in the control sample were positive. Concerning oral cancer development, it seems likely that HPV may be a cofactor, as 100% of patients who developed oral cancers within 4-12 years were all positive for HPV, one being positive for HPV 16.

Adult↗

Binding properties of Streptococcus gordonii SspA and SspB (antigen I/II family) polypeptides expressed on the cell surface of Lactococcus lactis MG1363.

The oral bacterium Streptococcus gordonii expresses two cell wall-associated polypeptides, designated SspA (1,542 amino acid residues) and SspB (1,462 amino acid residues), that have 70% sequence identity. These polypeptides are members of the antigen I/II family of oral streptococcal adhesins and mediate the binding of streptococci to salivary glycoproteins, collagen, and other oral microorganisms such as Actinomyces naeslundii. To determine if SspA and SspB have differential binding properties, the coding sequences of the sspA and sspB genes were cloned into expression plasmid vector pTREX1-usp45LS to generate pTREX1-sspA and pTREX1-sspB, respectively, and the Ssp polypeptides were displayed on the cell surface of Lactococcus lactis MG1363. Lactococcal cells expressing similar levels of surface SspA or SspB polypeptide were then compared for their abilities to adhere to a range of antigen I/II polypeptide substrates. More than twice as many L. lactis cells expressing SspA bound to immobilized salivary agglutinin glycoprotein (SAG) as did L. lactis cells expressing SspB. In contrast, lactococci expressing SspB adhered twice as well as lactococci producing SspA to collagen type I and to Candida albicans. The binding of A. naeslundii to lactococci was only weakly enhanced by surface expression of Ssp polypeptides. L. lactis(pTREX1-sspB) cells bound in greater numbers to SAG than did Enterococcus faecalis JH2-2 cells expressing SspB from pAM401EB-5. The results suggest that SspA and SspB have markedly different binding affinities for their oral substrates and thus may function to promote site diversity in colonization by S. gordonii.

Adhesins, Bacterial↗

Secretory immunoglobulin A in saliva of women with oral and genital HPV infection.

UNLABELLED: Secretory IgA contributes towards the protection of mucosal surfaces against invading microorganisms. OBJECTIVES: Quantify secretory IgA titers in the saliva of women with HPV in the oropharynx and/or in the genital area. SUBJECTS AND METHODS: Seventy women with clinical genital HPV lesions and 70 women without HPV infection were tested for oral HPV DNA and the levels of total IgA in their saliva. One millilitre of saliva was collected, centrifuged and stored at -80 degrees C for the measurement of secretory IgA by nephelometry technique. A pool of oral pharyngeal cells was collected for HPV identification by polymerase chain reaction. RESULTS: Oral HPV PCR was positive in 29 (21%) women (26 women with genital HPV and only 3 women without genital HPV). Titers of secretory IgA were extremely lower in-patients with HPV DNA in the oropharynx when compared to HPV negative women (p<0.0001). Genital HPV and smoking were also associated to low levels of total sIgA in saliva (p<0.01). After multivariable analyses only the presence of HPV in the oral cavity and/or in genital area, but not smoking, was related to low levels of total secretory IgA. CONCLUSION: Women with low levels of total secretory IgA could be more susceptible to having their oral mucosa colonized by HPV.

Adult↗

Monocyte chemoattractant protein 1 and interleukin-8 production in mononuclear cells stimulated by oral microorganisms.

Chemokines are a family of low-molecular-weight proinflammatory cytokines that stimulate recruitment of leukocytes. The chemokines interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1) are relatively specific chemoattractants for neutrophils and monocytes, respectively. Chemokine expression contributes to the presence of different leukocyte populations observed in normal and pathologic states. In the present studies, peripheral blood mononuclear cells (PBMC) were stimulated by microbes (Candida albicans, Streptococcus mutans, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans) selected based upon their importance as oral pathogens. IL-8 and MCP-1 gene expression and protein release were determined by Northern blot (RNA blot) analysis and enzyme-linked immunosorbent assay. C. albicans, P. gingivalis, and A. actinomycetemcomitans induced high levels of production of both MCP-1 and IL-8. S. mutans was a strong inducer of MCP-1, but it did not stimulate significant production of IL-8. C. albicans, S. mutans, and A. actinomycetemcomitans were 500 to 5,000 times more potent than P. gingivalis in terms of MCP-1 production. In general, the microbe-to-PBMC ratios required for maximum gene expression of MCP-1 were lower than those for IL-8. However, for actual protein release of MCP-1 versus IL-8, differences in the effects of various microbe concentrations were observed only for A. actinomycetemcomitans. These results demonstrate that different oral pathogens induce specific dose-dependent patterns of chemokine gene expression and release. Such patterns may help explain the immunopathology of oral infections, particularly with regard to inflammatory leukocyte recruitment.

Aggregatibacter actinomycetemcomitans↗

The recurrence of Helicobacter pylori infection: incidence and variables influencing it. A critical review.

The rate of H. pylori recurrence after eradication of the microorganism seems to be relatively low, at least in developed countries, where the mean annual reinfection rate is of approximately 3% per patient-year of follow-up, although the risk of reinfection in some developing regions is considerably higher. Several findings suggest that recrudescence rather than reinfection is likely to be responsible for most cases of recurrence: (i) the recurrences decrease with time and decline sharply after the first year, and (ii) studies using molecular fingerprinting techniques (polymerase chain reaction [PCR]) confirm that the identified microorganisms (before and after therapy) are usually genetically identical. The lower the efficacy of an antibiotic therapy, the greater the likelihood that recurrence occur, again suggesting that in these cases temporary "clearance" has been achieved rather than true eradication. The value of the (13)C-urea breath test after treatment is higher in those patients who suffer a recurrence; therefore, selection of a lower cut-off value may be helpful to maintain the diagnostic accuracy of posttreatment breath test, and thus preventing recrudescences. The observation of a pattern of histological (active) gastritis without the concomitant finding of H. pylori must raise the suspicion of a diagnostic error. Some studies suggest that recurrence is relatively infrequent, even if the patient's spouse is H. pylori-positive, suggesting that the patient's partner does not act as a reservoir for the reinfection. However, other investigators achieve contrary results, although a common exogenous source of H. pylori (for both partner's infection and patient's reinfection) cannot be ruled out. The oral cavity may be a potential source for recrudescence of gastric infection after successful therapy. When peptic ulcer reappears (sometimes with bleeding recurrence) or gastric MALT (mucosa-associated lymphoid tissue) lymphoma relapses after previous H. pylori eradication, recolonization of the gastric mucosa by the organism has almost always occurred.

Breath Tests↗

Effects of oral flora on platelets: possible consequences in cardiovascular disease.

During episodes of dental bacteremia, viridans group streptococci encounter platelets. Among these microorganisms, certain Streptococcus sanguis induce human and rabbit platelets to aggregate in vitro. In experimental rabbits, circulating streptococci induced platelets to aggregate, triggering the accumulation of platelets and fibrin into the heart valve vegetations of endocarditis. At necropsy, affected rabbit hearts showed ischemic areas. We therefore hypothesized that circulating S. sanguis might cause coronary thrombosis and signs of myocardial infarction (MI). Signs of MI were monitored in rabbits after infusion with platelet-aggregating doses of 4 to 40 x 10(9) cells of S. sanguis 133-79. Infusion resulted in dose-dependent changes in electrocardiograms, blood pressure, heart rate, and cardiac contractility. These changes were consistent with the occurrence of MI. Platelets isolated from hyperlipidemic rabbits showed an accelerated in vitro aggregation response to strain 133-79. Cultured from immunosuppressed children with septic shock and signs of disseminated intravascular coagulation, more than 60% of isolates of viridans streptococci induced platelet aggregation when tested in vitro. The data are consistent with a thrombogenic role for S. sanguis in human disease, contributing to the development of the vegetative lesion in infective endocarditis and a thrombotic mechanism to explain the additional contributed risk of periodontitis to MI.

Animals↗

In vitro evaluation of virulence attributes of Candida spp. isolated from patients affected by diabetes mellitus.

BACKGROUND: Diabetes mellitus is a common disease found worldwide and it has been previously suggested that oral candidal infections may be more frequent or severe in patients with this disease. Systemic and local factors may influence the balance between the host and yeasts, and favour the transformation of Candida isolates from commensal to pathogenic microorganisms. Candida species have developed specific virulence mechanisms that confer the ability to colonise host surfaces, to invade deeper host tissue, or to evade host defences. Few studies have investigated the expression of the virulence attributes of oral Candida isolates in patients with diabetes mellitus. MATERIAL AND METHODS: The in vitro extracellular proteinase production and the in vitro ability to adhere to fibronectin of 229 Candida isolates of two geographic different groups of patients with diabetes mellitus and of healthy subjects were assessed. RESULTS: Candida isolates of patients with diabetes mellitus expressed a higher ability to adhere than those of healthy subjects. Higher levels of adhesion were also recorded in patients with a lower oral Candida colonisation. No differences were observed in the in vitro expression of extracellular proteinase of Candida isolates of patients with diabetes mellitus and those of non-diabetic subjects. Isolates of patients with type 2 diabetes mellitus expressed greater levels of proteinase than isolates of type 1 diabetes mellitus. CONCLUSIONS: Diabetes mellitus could be considered as an additional variable that may influence not only oral Candida carriage but also the ability of isolates to enhance the expression of virulence attributes.

Candida↗

Endocardiac infectivity and binding to extracellular matrix proteins of oral Abiotrophia species.

Microorganisms of the genus Abiotrophia, formerly known as nutritionally variant streptococci, are members of the oral flora and often isolated from patients with endocarditis, but pathogenicity of oral Abiotrophia species has not been examined yet. In this study, 17 strains isolated from healthy human oral cavities and 7 reference strains (all derived from patients with endocarditis) of Abiotrophia spp. were tested for their abilities to cause infections in damaged heart tissues in catheterized rats and to adhere to extracellular matrix proteins in vitro. The reference strains of A. defectiva and A. adiacens showed high infectivities in the rats. Four oral isolates of these two species showed similarly high infectivities and three had moderate infectivities. Most of 10 oral strains of A. para-adiacens and A. elegans were found to be generally less infective. The highly infective A. adiacens strains showed markedly high fibronectin-binding capacity, suggesting a possible relationship between the fibronectin-binding capacity and damaged heart tissue infectivity of the Abiotrophia species. A. defectiva strains which were also highly infective had moderate levels of binding to fibronectin and other extracellular matrix proteins. Most of A. para-adiacens and A. elegans strains showed low or negligible binding capacities to any extracellular matrix proteins tested.

Animals↗

Microbial biofilms: from ecology to molecular genetics.

Biofilms are complex communities of microorganisms attached to surfaces or associated with interfaces. Despite the focus of modern microbiology research on pure culture, planktonic (free-swimming) bacteria, it is now widely recognized that most bacteria found in natural, clinical, and industrial settings persist in association with surfaces. Furthermore, these microbial communities are often composed of multiple species that interact with each other and their environment. The determination of biofilm architecture, particularly the spatial arrangement of microcolonies (clusters of cells) relative to one another, has profound implications for the function of these complex communities. Numerous new experimental approaches and methodologies have been developed in order to explore metabolic interactions, phylogenetic groupings, and competition among members of the biofilm. To complement this broad view of biofilm ecology, individual organisms have been studied using molecular genetics in order to identify the genes required for biofilm development and to dissect the regulatory pathways that control the plankton-to-biofilm transition. These molecular genetic studies have led to the emergence of the concept of biofilm formation as a novel system for the study of bacterial development. The recent explosion in the field of biofilm research has led to exciting progress in the development of new technologies for studying these communities, advanced our understanding of the ecological significance of surface-attached bacteria, and provided new insights into the molecular genetic basis of biofilm development.

Bacterial Adhesion↗

Inhibitory effect of sodium fluoride and chlorhexidine on the growth of oral lactobacilli.

The accumulation of microorganisms in dental plaque is related to the etiology of caries and periodontal disease, with a high prevalence worldwide. The prophylactic measures include the use of chemical agents as NaF and chlorhexidine. Lactic acid bacteria are members of the normal microbiota of the oral cavity being discussed with regard to their beneficial or detrimental effect in this environment. The present study was performed to determine the growth of some species of Lactobacillus at different concentrations of NaF and chlorhexidine. The strains were isolated from both caries-free and caries patients. Their growth parameters were evaluated by the application of the Gompertz model to the experimental data of optical density as a measurement of growth. The degree of inhibition of the growth of all of the lactobacilli studied was different, depending on each particular strain. NaF at 1 mmol x L(-1) inhibited between 5% and 46%, at 5 mmol x L(-1) between 13% and 65%, and at 20 mmol x L(-1) between 57% and 84%. CHX at higher concentrations (197 and 98 mmol x L(-1) showed a complete inhibition of some of the strains. The significance of the results was evaluated by the application of a multivariate analysis and also compared with the inhibition of pathogenic Streptococcus mutans and with lactobacilli strains from collection cultures.

Chlorhexidine↗

Human beta-defensins 2 and 3 demonstrate strain-selective activity against oral microorganisms.

Human beta-defensins 2 and 3 (HBD-2 and HBD-3) are inducible peptides present at sites of infection in the oral cavity. A few studies have reported broad-spectrum antimicrobial activity for both peptides. However, no comprehensive study has thoroughly investigated their potential against oral pathogens. The purpose of this study was to test the effectiveness of HBD-2 and HBD-3 against a collection of oral organisms (Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, Peptostreptococcus micros, Actinomyces naeslundii, Actinomyces israelii, Streptococcus sanguis, Streptococcus mutans, Candida tropicalis, Candida parapsilosis, Candida krusei, Candida glabrata, and Candida albicans). Radial diffusion assays were used to test HBD-2 and HBD-3 activities against at least three strains of each species. There was significant variability in MICs, which was strain specific rather than species specific. MICs ranged from 3.9 to >250 micro g/ml for HBD-2 and from 1.4 to >250 micro g/ml for HBD-3. HBD-3 demonstrated greater antimicrobial activity and was effective against a broader array of organisms. Overall, aerobes were 100% susceptible to HBD-2 and HBD-3, whereas only 21.4 and 50% of the anaerobes were susceptible to HBD-2 and HBD-3, respectively. HBD-2 and HBD-3 also demonstrated strain-specific activity against the Candida species evaluated. Interestingly, an association between HBD-2 and HBD-3 activities was noted. This suggests that the two peptides may have similar mechanisms yet utilize distinct pathways. The lack of activity against specific anaerobic strains and Candida warrants further investigation of the potential resistance mechanisms of these organisms. Finally, the significant variability between strains underlies the importance of testing multiple strains when evaluating activities of antimicrobial peptides.

Anti-Infective Agents↗

The efficacy of topical antibiotic prophylaxis for contaminated head and neck surgery.

This study was undertaken to determine the feasibility of using perioperative topical antibiotics in contaminated head and neck surgery and to standardize the culture methodology (both qualitative and quantitative) which could serve as bacteriologic endpoints for evaluation. Following preliminary studies to establish oral cavity indicator organisms and the impact of a single antibiotic mouthwash dose on oral microflora, 10 consecutive patients undergoing contaminated head and neck surgery were recruited into a clinical trial where clindamycin mouthwash and intraoperative irrigation containing clindamycin were used instead of traditional parenteral antibiotics. The bacteriologic efficacy of topical clindamycin was assessed by comparing the presence of four indicator microorganisms (two aerobic and two anaerobic) cultured from two oral cavity culture sites before and after antibiotic prophylaxis. The patients included in the study underwent total laryngectomy plus neck dissection(s) for laryngeal or hypopharyngeal carcinoma from 1991 to 1992 at a large university hospital specializing in head and neck cancer surgery. The main outcome measures used were the development of a postoperative wound infection and quantitative and qualitative bacteriology of the intraoperative neck wound and postoperative oral cavity. Two aerobic and two anaerobic organisms proved useful as a practical indicator for bacteriologic efficacy. Preoperative mouthwash resulted in a 99% reduction of both aerobic and anaerobic bacteria in intraoperatively cultured neck sites. Irrigation during surgery with the clindamycin solution further reduced the bacterial neck counts by an additional 90%. There was a consistent overgrowth of Hemophilus species on postoperative oral cavity cultures. No patient developed a postoperative wound infection. A topical prophylactic antibiotic alone was efficacious and safe for patients undergoing major contaminated head and neck surgery. Culture methods for assessment of bacteriologic efficacy were reproducible and cost-effective. This pilot study furnishes the ethical and scientific basis for large-scale prospective trials comparing topical versus parenteral antimicrobial agents.

Administration, Topical↗

Electrophoretic protein patterns and numerical analysis of Candida albicans from the oral cavities of healthy children.

The aim of this research was to evaluate the protein polymorphism degree among seventy-five C. albicans strains from healthy children oral cavities of five socioeconomic categories from eight schools (private and public) in Piracicaba city, São Paulo State, in order to identify C. albicans subspecies and their similarities in infantile population groups and to establish their possible dissemination route. Cell cultures were grown in YEPD medium, collected by centrifugation, and washed with cold saline solution. The whole-cell proteins were extracted by cell disruption, using glass beads and submitted to SDS-PAGE technique. After electrophoresis, the protein bands were stained with Coomassie-blue and analyzed by statistics package NTSYS-pc version 1.70 software. Similarity matrix and dendrogram were generated by using the Dice similarity coefficient and UPGMA algorithm, respectively, which made it possible to evaluate the similarity or intra-specific polymorphism degrees, based on whole-cell protein fingerprinting of C. albicans oral isolates. A total of 13 major phenons (clusters) were analyzed, according to their homogeneous (socioeconomic category and/or same school) and heterogeneous (distinct socioeconomic categories and/or schools) characteristics. Regarding to the social epidemiological aspect, the cluster composition showed higher similarities (0.788 < SD < or = 1.0) among C. albicans strains isolated from healthy children independent of their socioeconomic bases (high, medium, or low). Isolates of high similarity were not found in oral cavities from healthy children of social stratum A and D, B and D, or C and E. This may be explained by an absence of a dissemination route among these children. Geographically, some healthy children among identical and different schools (private and public) also are carriers of similar strains but such similarity was not found among other isolates from children from certain schools. These data may reflect a restricted dissemination route of these microorganisms in some groups of healthy scholars, which may be dependent of either socioeconomic categories or geographic site of each child. In contrast to the higher similarity, the lower similarity or higher polymorphism degree (0.499 < or = SD < 0.788) of protein profiles was shown in 23 (30.6%) C. albicans oral isolates. Considering the social epidemiological aspect, 42.1%, 41.7%, 26.6%, 23.5%, and 16.7% were isolates from children concerning to socioeconomic categories A, D, C, B, and E, respectively, and geographically, 63.6%, 50%, 33.3%, 33.3%, 30%, 25%, and 14.3% were isolates from children from schools LAE (Liceu Colégio Albert Einstein), MA (E.E.P.S.G. "Prof. Elias de Melo Ayres"), CS (E.E.P.G. "Prof. Carlos Sodero"), AV (Alphaville), HF (E.E.P.S.G. "Honorato Faustino), FMC (E.E.P.G. "Prof. Francisco Mariano da Costa"), and MEP (E.E.P.S.G. "Prof. Manasses Ephraim Pereira), respectively. Such results suggest a higher protein polymorphism degree among some strains isolated from healthy children independent of their socioeconomic strata or geographic sites. Complementary studies, involving healthy students and their families, teachers, servants, hygiene and nutritional habits must be done in order to establish the sources of such colonization patterns in population groups of healthy children. The whole-cell protein profile obtained by SDS-PAGE associated with computer-assisted numerical analysis may provide additional criteria for the taxonomic and epidemiological studies of C. albicans.

Brazil↗

Superinfecting microorganisms in patients under treatment with cyclosporin-A and its correlation to gingival overgrowth.

The aim of this study was to identify the presence of superinfecting microorganisms (Gram-negative enteric rods and Candida sp.) in heart transplant patients and correlate this with gingival overgrowth. Thirty patients (10 females, 20 males--mean age 45 years) were examined. All were under cyclosporin-A (CsA) therapy. Patients who had taken any antibiotics 3 months prior the study or had been submitted to periodontal therapy were not enrolled. Patients were required to have at least 6 teeth. The plaque index (PI), gingival index (GI), pocket depth (PD) and clinical attachment level (CAL) were recorded. Microbiological samples were taken from sulcus/pocket (s/p) and from stimulated saliva (ss) and submitted to analysis. Patients were divided into two groups: the ones with gingival overgrowth (GO) and those without gingival overgrowth (WGO). After statistical analysis (chi-square test, Student's t-test, Fisher test, p < or = 0.05), we concluded that there was no statistical difference between groups in the parameters of gender, CsA dosage, time since transplantation, PI, GI, PD and CAL. Gram-negative rods from either the sulcus/pocket or saliva samples were not found. Candida sp. was detected (s/p-30% and ss-30%). Stimulated saliva samples analysis determined that the presence of Candida sp. was associated with patients without gingival overgrowth.

Adolescent↗

Depth profiling of the elemental surface composition of the oral microorganism S. salivarius HB and fibrillar mutants by X-ray photoelectron spectroscopy.

X-ray photoelectron spectroscopy (XPS) on microbial cell surfaces requires freeze-drying of cells, and as a result, the cell surface appendages flatten out on the cell surface and form a collapsed fibrillar mass. At present, it is unclear how the density, length and composition of these fibrils influence the elemental surface composition as probed by XPS. The sampling depth of XPS can be varied by changing the electron take-off angle. In this article, we made a depth profiling of the collapsed fibrillar mass of Streptococcus salivarius HB and fibril-deficient mutants by angle-dependent XPS. Methylamine tungstate negative staining and ruthenium red staining followed by sectioning revealed distinct classes of fibrils with various lengths on each of the strains. Interpretation of the angle dependence of the oxygen/carbon (O/C) and phosphorus/carbon (P/C) surface concentration ratios of these strains was difficult. However, the angle dependence of the nitrogen/carbon (N/C) surface concentration ratio could be fully interpreted: N/C did not vary with sampling depth on a bald strain, S. salivarius HBC12 and on S. salivarius HB7, a strain with a dense array of fibrils of uniform length. N/C decreased with sampling depth in case of a sparsely fibrillated strain, S. salivarius HBV51 and eventually reached the value observed for the bald strain, HBC12. A high N/C at small sampling depth was observed for S. salivarius HB with protruding, protein rich fibrils. We conclude that elemental depth profiling of microbial cell surfaces by XPS can be interpreted to coincide with structural and biochemical information on the cell surface as obtained by electron microscopy and can therefore be considered as a useful technique to study structural features of cell surfaces in combination with electron microscopy.

Freeze Drying↗

[Pharmacokinetic/pharmacodynamic analysis of antibiotic therapy in dentistry and stomatology].

INTRODUCTION: This study evaluates the efficacy of various antimicrobial treatments for orofacial infections on the basis of pharmacokinetic/pharmacodynamic (PK/PD) criteria. METHODS: A complete a literature search was undertaken to establish the MIC90 values of the five microorganisms most frequently isolated in odontogenic infections and the pharmacokinetic parameters of 13 antibiotics used in these infections. Pharmacokinetic simulations were then carried out with mean population parameters and efficacy indexes were calculated for the 47 treatment regimens analyzed. For drugs showing time-dependent antibacterial killing, the time above MIC (t > MIC) was calculated. For drugs with concentration-dependent bactericidal activity, the AUC/MIC was calculated. RESULTS: Amoxicillin-clavulanic (500 mg/8 h or 1000 mg/12 h) and clindamycin (300 mg/6 h) in the time-dependent killing group and moxifloxacin (400 mg/24 h) in the concentration-dependent group showed adequate efficacy indexes against the five pathogens considered to be the most commonly implicated in odontogenic infections. The spiramycin plus metronidazole combination, present in the commercial formulation Rhodogyl, did not reach satisfactory PK/PD indexes. CONCLUSION: PK/PD indexes, which are useful predictors of the potential efficacy of antibacterial therapy, were used with ontogenic infections in the present study. The PK/PD simulations showed that amoxicillin-clavulanic, clindamycin and moxifloxacin were the most suitable antibiotics for this kind of infection. Clinical trials are required to confirm that this methodology is useful in these pathologic processes.

Algorithms↗

Involvement of antigen I/II surface proteins in Streptococcus mutans and Streptococcus intermedius biofilm formation.

BACKGROUND/AIM: Dental diseases are caused by microorganisms organized in biofilms. Streptococcus mutans and Streptococcus intermedius are commensals of the human oral cavity. S. mutans is associated with caries, whereas S. intermedius is associated with purulent infections. Oral streptococci including S. mutants and S. intermedius express a family of surface proteins termed antigen I/II (Ag I/II). Ag I/II is implicated in adhesion; however, its role in biofilm formation has not yet been investigated. METHODS: By using isogenic Ag I/II-deficient mutants of S. mutans and S. intermedius we studied the influence of Ag I/II on in vitro biofilm formation. Biofilm was quantified in polystyrene microtiter plates and visualized by scanning electron microscopy. Ag I/II expression in planktonic and biofilm cells, as well as in the presence or absence of saliva was investigated by immunoblotting. RESULTS: In the presence of saliva, the Ag I/II-deficient mutants formed 65% less biofilm than the wild-types. In the absence of saliva, no difference was observed in S. mutans, whereas the S. intermedius Ag I/II mutant formed 41% less biofilm. Ag I/II expression was reduced in the presence of saliva. No differences in expression were observed between biofilm and planktonic cells. CONCLUSION: The results indicated that Ag I/II may be important during biofilm formation particularly in the presence of saliva. These findings may provide useful information regarding the importance of Ag I/II in biofilm formation and in the search of new strategies to control biofilm-mediated infections.

Bacterial Adhesion↗

Differential recognition of oral indigenous bacteria by salivary immunoglobulins A and G.

Assuming that salivary immunity to indigenous microorganisms could develop, we assessed antibacterial reactivities of natural salivary antibodies in specific pathogen-free inbred mice. An ELISA was set up, using whole bacterial cells, to map reactivities of salivary IgA and IgG which accounted respectively for 91% and 8.7% of salivary Ig's in the BALB/c mouse. Representative strains of seven species from three genera (Lactobacilli, Staphylococci, and Streptococci), including major and minor components of the murine oral flora (38, 43, and 8%, respectively), were used to determine the presence and level of specific antibodies in individual saliva. It was verified that naturally occurring IgA antibodies can display diverse antibacterial reactivities. A characteristic profile emerged for salivary IgA where antibodies to Streptococcus faecalis predominate. Natural salivary IgG antibodies did not show the same reactivity pattern as IgA, anti-Lactobacilli and anti-Staphylococci reactivities being much less frequent in the salivary IgG repertoire. However, antibodies to S. faecalis occurred at the same high frequency for both isotypes (62-70% of the samples). Besides being species-specific, antibacterial reactivities were also found to be strain-specific. Broad variations in antibacterial titers were detected among individual mice under standardized experimental conditions. Present data thus suggest that the dynamics of salivary antibody production in the mouse reflect a differential natural sensitization of the secretory (IgA) versus the systemic (IgG) immune systems by distinct populations of indigenous bacteria.

Animals↗