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Dental caries is a multifactorial disease driven by interactions between the highly complex microbial biofilm community and host factors like diet, oral hygiene habits, and age. The oral streptococci are one of the most dominant members of the plaque biofilm and are implicated in disease but also in maintaining oral health. Current methods used for studying the supragingival plaque community commonly sequence portions of the16S rRNA gene, which often cannot taxonomically resolve members of the streptococcal community past the genus level due to their sequence similarity. The goal of this study was to design and evaluate a more reliable and cost-effective method to identify oral streptococci at the species level by applying a new locus, the 30S-S11 rRNA gene, for high-throughput amplicon sequencing. The study results demonstrate that the newly developed single-copy 30S-S11 gene locus resolved multiple amplicon sequence variants (ASVs) within numerous species, providing much improved taxonomic resolution over 16S rRNA V4. Moreover, the results reveal that different ASVs within a species were found to change in abundance at different stages of caries progression. These findings suggest that strains of a single species may perform distinct roles along a biochemical spectrum associated with health and disease. The improved identification of oral streptococcal species will provide a better understanding of the different ecological roles of oral streptococci and inform the design of novel oral probiotic formulations for prevention and treatment of dental caries. IMPORTANCE The microbiota associated with the initiation and progression of dental caries has yet to be fully characterized. Although much insight has been gained from 16S rRNA hypervariable region DNA sequencing, this approach has several limitations, including poor taxonomic resolution at the species level. This is particularly relevant for oral streptococci, which are abundant members of oral biofilm communities and major players in health and caries disease. Here, we develop a new method for taxonomic profiling of oral streptococci based on the 30S-S11 rRNA gene, which provides much improved resolution over 16S rRNA V4 (resolving 10 as opposed to 2 species). Importantly, 30S-S11 can resolve multiple amplicon sequence variants (ASVs) within species, providing an unprecedented insight into the ecological progression of caries. For example, our findings reveal multiple incidences of different ASVs within a species with contrasting associations with health or disease, a finding that has high relevance toward the informed design of prebiotic and probiotic therapy.

The goal of restoring caries lesions is to protect the pulp, prevent progression of the disease process, and restore the form and function of the tooth. The purpose of this systematic review was to determine the effect of different direct restorative materials for treating cavitated caries lesions on anterior and posterior primary and permanent teeth.The authors included parallel and split-mouth randomized controlled trials comparing the effectiveness of direct restorative materials commercially available in the United States placed in vital, nonendodontically treated primary and permanent teeth. Pairs of reviewers independently conducted study selection, data extraction, and assessments of risk of bias and certainty of the evidence using the Grading of Recommendations Assessment, Development and Evaluation approach. The authors conducted pair-wise meta-analyses to summarize the evidence and calculated measures of association and their 95% CIs.Thirty-eight randomized controlled trials were eligible for analysis, which included data on Class I and Class II restorations on primary teeth and Class I, Class II, Class III, Class V, and root surface restorations on permanent teeth. Included studies assessed the effect of amalgam, resin composite, compomer, conventional glass ionomer cement, resin-modified glass isomer cement, and preformed metal crowns. Moderate to very low certainty evidence suggested varying levels of effectiveness across restorative materials.Owing to a relatively low event rate across various outcomes indicating restoration failure, there was limited evidence to support important differences between direct restorative materials used in practice.

Clinical Relevance Radiographic lesion depth should not be used as the single determinant of the restorative threshold for clinically inaccessible approximal caries lesions. Temporary tooth separation is a feasible and effective diagnostic aid for assessment and appropriate management of approximal lesions. SUMMARY In the era of tooth-preserving dentistry, the decision to restore approximal caries lesions must be based on the accurate assessment of tooth cavitation, as the accumulation of oral biofilms in these areas encourages lesion progression. However, lesions radiographically into dentin remain the main threshold criterion for restoring approximal lesions even though most of these lesions may not be cavitated. A school-based clinical protocol for temporary tooth separation (TTS) was developed to improve visual-tactile assessment and management of clinically inaccessible approximal lesions. TTS data retrieved from electronic health records were used to correlate radiographic lesion depth and surface cavitation status with lesion location and the patient’s caries risk and to evaluate the effectiveness of TTS as a diagnostic aid for approximal lesions. Of the 206 lesions assessed, 66.5% (n=137) were located in the maxillary arch, 56.6% (n=116) in distal surfaces, 61.3% (n=114) in premolars, and 21.5% (n=40) in molars. After tooth separation, 79.6% (n=164) of the lesions were diagnosed as noncavitated, including 90% (n=66) of the lesions radiographically at the inner half of enamel (E2) and 66% (n=49) of those at the outer-third of dentin (D1). Logistic regression analysis using E2 and D1 lesions showed no significant association between lesion depth or cavitation status with lesion location and caries risk. TTS is a feasible and effective diagnostic aid for the assessment and appropriate management of approximal caries lesions. There is a need to reevaluate the use of radiographic lesion depth as the single determinant of the restorative threshold for clinically inaccessible approximal lesions.

(1) Background: The amino acid arginine is now receiving great attention due to its potential anti-caries benefits. The purpose of this in vitro study was to evaluate the shear bond strength (SBS), ultimate tensile strength (UTS), and antimicrobial potential (CFU) of two arginine-containing orthodontic resin cements. (2) Methods: Forty bovine incisors were separated into four groups (n = 10): Orthocem, Orthocem + arginine (2.5 wt%), Transbond XT, and Transbond XT + arginine (2.5 wt%). The brackets were fixed to the flat surface of the enamel, and after 24 h the SBS was evaluated using the universal testing machine (Instron). For the UTS test, hourglass samples (n = 10) were made and tested in a mini-testing machine (OM-100, Odeme). For the antibacterial test (colony forming unit-CFU), six cement discs from each group were made and exposed to Streptococcus mutans UA159 biofilm for 7 days. The microbiological experiment was performed by serial and triplicate dilutions. The data from each test were statistically analyzed using a two-way ANOVA, followed by Tukey’s test (α = 0.05). (3) Results: The enamel SBS mean values of Transbond XT were statistically higher than those of Orthocem, both with and without arginine (p = 0.02033). There was no significant difference in the SBS mean values between the orthodontic resin cements, either with or without arginine (p = 0.29869). The UTS of the Transbond XT was statistically higher than the Orthocem, but the addition of arginine at 2.5 wt% did not influence the UTS for either resin cement. The Orthocem + arginine orthodontic resin cement was able to significantly reduce S. mutans growth, but no difference was observed for the Transbond XT (p = 0.03439). (4) Conclusion: The incorporation of arginine to commercial orthodontic resin cements may be an efficient preventive strategy to reduce bacterial growth without compromising their adhesive and mechanical properties.

Dental caries is closely related to a dysbiosis of the microbial consortia of supragingival oral biofilms driven by a sugar-frequent and acidic-pH environment. The pH is a key factor affecting the homeostasis of supragingival biofilms seen in health. There is increasing interest on the ecological dynamics of the oral microbiome and how a dysbiotic microbiota can be successfully replaced by health-beneficial flora. The concept of preventing the microbial dysbiosis related to caries through modulation of sugar intake and pH has fully emerged.

INTRODUCTION: Ammonia production via the arginine deiminase system (ADS) of oral bacteria can function to reduce the cariogenicity of oral biofilms by neutralizing glycolytic acids that cause tooth demineralization. OBJECTIVES: This cohort study investigated the relationship between ADS activity and bacterial profile changes of supragingival biofilms with caries experience among children over time. METHODS: A total of 79 children aged 2 to 7 y at baseline were assessed every 6 mo for a period of 18 mo. Children were grouped as caries free (CF), caries active with enamel lesions (CAE), or caries active with dentin lesions (CA). Supragingival plaque samples were collected from caries-free surfaces (PF) and from enamel (PE) and dentin (PD) lesions. Plaque ADS activity was measured by monitoring citrulline production from arginine and compared with ribosomal 16S rRNA–derived taxonomic profiles for the same samples. RESULTS: At baseline, 37% of the children were CF, 34% CAE, and 29% CA. At 18 mo, 26% were CF, 41% CAE, 23% CA, and 10% were caries experienced (new restorations but no caries activity). Throughout the study period, ADS activity was significantly higher in the CF group than the CA group (P < 0.0001), and ADS activity in the PF samples was significantly higher than in the PE and PD samples (P < 0.0001). Distance-based redundancy analysis showed that the bacterial communities could be differentiated when plaque samples are grouped into levels of high and low ADS activity. CONCLUSIONS: There is a positive correlation between caries activity and low arginolytic capacity of the supragingival oral biofilms of children and tooth surfaces over time. Measurements of arginine metabolism via ADS may be useful to differentiate the caries risk of individuals and tooth surfaces. KNOWLEDGE TRANSFER STATEMENT: Findings from this study support the development of new strategies for caries risk assessment and prevention based on modulation of the virulence of the oral microbiome through arginine metabolism in supragingival biofilms.

Introduction: Dental caries is a chronic complex disease of multifactorial etiology that affects a quarter of U.S. children. This study evaluated the association between prenatal smoking and offspring caries experience and used a negative control exposure analysis to assess if the association is causal. Methods: Data from 1429 mother-offspring participants of the 1991/92 Avon Longitudinal Study of Parents and Children conducted in Bristol, England were analyzed. Prenatal smoking (yes v. no) and quantity smoked (none

Dental caries is one of the most common diseases worldwide. Bacteria and fungi are both commensals in the oral cavity; however, most research regarding caries has focused on bacterial impacts. The oral fungal mycobiome associated with caries is not well characterized, and its role in disease is unclear. ITS1 amplicon sequencing was used to generate taxonomic profiles from site-specific supragingival plaque samples (n = 82) obtained from 33 children with different caries status. Children were either caries free (CF), caries active with enamel lesions (CAE), or caries active with dentin lesions (CA). Plaque samples were collected from caries-free surfaces (PF) and from enamel (PE) and dentin (PD) lesions. Taxonomic profiles representing the different categorizations (CF-PF, CAE-PF, CAE-PE, CA-PF, CA-PE, and CA-PD) were used to characterize the mycobiome and its change through disease progression. A total of 139 fungal species were identified. Candida albicans was the most abundant species, followed by Candida dubliniensis. We found that severely progressed plaque communities (CA-PD) were significantly different from healthy plaque communities (CF-PF). A total of 32 taxa were differentially abundant across the plaque categories. C. albicans, C. dubliniensis, Nigrospora oryzae, and an unclassified Microdochium sp. were correlated with caries, whereas 12 other taxa were correlated with health. C. dubliniensis increased steadily as caries progressed, suggesting that C. dubliniensis may play an important role in caries pathogenicity. In contrast, four health-associated fungal taxa have the potential to antagonize the cariogen Streptococcus mutans via xylitol production, suggesting a possible fungal mechanism that could contribute to maintenance of dental health. IMPORTANCE Early-childhood caries is one of the most prevalent diseases in children worldwide and, while preventable, remains a global public health concern. Untreated cavities are painful and expensive and can lead to tooth loss and a lower quality of life. Caries are driven by acid production via microbial fermentation of dietary carbohydrates, resulting in enamel erosion. While caries is a well-studied disease, most research has focused on bacterial impacts, even though fungi are commensal organisms living within the plaque biofilm. There is very little known about how fungi impact caries pathogenicity. The elucidation of fungal taxa involved in caries disease progression is necessary for a more holistic view of the human oral microbiome. Data from this study will improve our understanding of how the fungal community changes as disease progresses and provide insight into the complex etiology of dental caries, which is necessary for the development of treatment plans and preventative measures.

Pre-existing maternal overweight/obesity and pregnancy weight gain are associated with adverse birth outcomes such as low birth weight and prematurity, which may increase the risk of developmental tooth defects and early childhood caries. We sought to investigate the association between prepregnancy BMI, gestational weight gain (GWG) and the risk of early childhood caries. Data from 1,429 mother-offspring participants of the 1991/1992 Avon Longitudinal Study of Parents and Children were analyzed. The exposures were prepregnancy BMI (under/normal weight vs. overweight/obese), and gestational weight gain (GWG) based on the Institute of Medicine’s recommended levels. The main outcome measured was offspring caries experience determined by clinical oral examinations at three time points. Log binomial regression estimated risk ratios and 95% confidence intervals. Seventy six percent (76%) of the mothers were under/normal weight prepregnancy, 39% and 26% respectively gained less and more than the recommended weight for their prepregnancy BMI during pregnancy. Being overweight/obese prepregnancy was associated with unadjusted RR (95% CI) of offspring caries of 1.16 (0.90, 1.51) at 31-months, 1.20 (0.96, 1.49) at 43-months and 1.09 (0.91, 1.30) at 61-months. GWG less than recommended was associated with higher unadjusted offspring caries experience of 1.13 (0.86, 1.48), 1.17 (0.92, 1.48) and 1.04 (0.87, 1.25) at 31-months, 43-months and 61-months respectively. There was insufficient evidence to indicate an association between prepregnancy BMI and gestational weight gain on offspring caries experience risk.

The goal of nonrestorative or non- and microinvasive caries treatment (fluoride- and nonfluoride-based interventions) is to manage the caries disease process at a lesion level and minimize the loss of sound tooth structure. The purpose of this systematic review and network meta-analysis was to summarize the available evidence on nonrestorative treatments for the outcomes of 1) arrest or reversal of noncavitated and cavitated carious lesions on primary and permanent teeth and 2) adverse events. We included parallel and split-mouth randomized controlled trials where patients were followed for any length of time. Studies were identified with MEDLINE and Embase via Ovid, Cochrane CENTRAL, and Cochrane Database of Systematic Reviews. Pairs of reviewers independently conducted the selection of studies, data extraction, risk-of-bias assessments, and assessment of the certainty in the evidence with the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach. Data were synthesized with a random effects model and a frequentist approach. Forty-four trials (48 reports) were eligible, which included 7,378 participants and assessed the effect of 22 interventions in arresting or reversing noncavitated or cavitated carious lesions. Four network meta-analyses suggested that sealants + 5% sodium fluoride (NaF) varnish, resin infiltration + 5% NaF varnish, and 5,000-ppm F (1.1% NaF) toothpaste or gel were the most effective for arresting or reversing noncavitated occlusal, approximal, and noncavitated and cavitated root carious lesions on primary and/or permanent teeth, respectively (low- to moderate-certainty evidence). Study-level data indicated that 5% NaF varnish was the most effective for arresting or reversing noncavitated facial/lingual carious lesions (low certainty) and that 38% silver diamine fluoride solution applied biannually was the most effective for arresting advanced cavitated carious lesions on any coronal surface (moderate to high certainty). Preventing the onset of caries is the ultimate goal of a caries management plan. However, if the disease is present, there is a variety of effective interventions to treat carious lesions nonrestoratively.

Predoctoral dental psychomotor examinations are known to generate high levels of stress among dental students, which may compromise their academic performance. At one U.S. dental school, all 93 first-year dental students were invited to attend a series of three workshop sessions prior to enrollment in their initial operative dentistry course. The workshops were developed to facilitate academic transition from the dental anatomy course to the operative dentistry course; provide early exposure to materials, instruments, and laboratory techniques; support the early development of psychomotor and self-assessment skills; and lessen students' stress and anxiety levels regarding psychomotor examinations. The aim of this study was to assess the impact of the workshops on the students' academic performance and self-reported stress and preparedness. All students who attended the workshop sessions and all who did not were asked to complete a pre-exam survey (immediately preceding the exam) and a post-exam survey (immediately after the exam) on the day of their first operative dentistry psychomotor exam. Of the 93 students, 21 attended one, 34 attended two, and 25 attended three workshop sessions, while 13 students did not attend any. Response rates for the pre- and post-exam surveys were 100% and 98.9%, respectively. Students who attended all three workshop sessions reported being significantly less stressed about taking the exam than the other groups. The mean exam grade of students who attended the workshop sessions was significantly higher than that of students who did not attend the sessions. These findings support the development and implementation of preparatory workshops to improve academic performance and decrease the stress levels of dental students prior to the first operative dentistry psychomotor exam.

Arginine metabolism via the arginine deiminase system (ADS) of oral bacteria generates ammonia, which can increase the pH of oral biofilms and decrease the risk for dental caries. Antagonistic interactions between ADS-positive and cariogenic bacteria in oral biofilms may be an important ecological determinant of caries. This study investigated the antagonistic potential and mechanisms of clinical isolates of arginolytic streptococci on and by Streptococcus mutans UA159, a well-characterized cariogenic human isolate. Low-passage isolates of Streptococcus gordonii, Streptococcus sanguinis, Streptococcus parasanguinis, Streptococcus australis, and Streptococcus cristatus inhibited the growth of S. mutans to various degrees when they were inoculated on growth media first or simultaneously with S. mutans. The antagonistic effects of arginolytic strains against S. mutans and the production of H2O2 by these strains were enhanced during growth in a less-rich medium or when galactose was substituted for glucose as the primary carbohydrate source. Pyruvate oxidase was the dominant pathway for H2O2 production by arginolytic strains, but lactate oxidase activity was also detected in some strains of S. gordonii and S. cristatus. UA159 inhibited the growth of all tested arginolytic strains when inoculated first, especially in aerobic conditions. However, the antagonistic effects of S. mutans on certain strains of S. gordonii and S. australis were not observed during anaerobic growth in the presence of arginine. Thus, arginolytic commensal streptococci may have a synergistically positive impact on the ecology of oral biofilms by moderating biofilm pH while antagonizing the growth and virulence of caries pathogens.

Secondary caries at the margins of composite restorations has been attributed to adhesive failure and consequent accumulation of cariogenic biofilms. Objectives To develop and evaluate an etch-and-rinse adhesive system containing arginine for sustainable release and recharge without affecting its mechanical properties. Arginine metabolism by oral bacteria generates ammonia, which neutralizes glycolytic acids and creates a neutral environmental pH that is less favorable to the growth of caries pathogens, thus reducing the caries risk at the tooth-composite interface. Methods Experimental adhesives were formulated with methacrylate monomers and arginine at 5%, 7%, and 10% or no arginine (control). Adhesives were tested for: (i) mechanical properties of true stress (FS and UTS), modulus of elasticity (E), degree of conversion (DC), Knoop hardness number (KHN) and dentin microtensile bond strength (μ-TBS), (ii) arginine release and recharge, and (iii) antibacterial activities. Data was analyzed by t -test, one-way ANOVA and Tukey’s tests. Results FS and UTS results showed no statistically significant differences between the 7% arginine-adhesive and control, while the results for E, DC, KHN and μ-TBS showed no difference among all groups. The 7% arginine-adhesive showed a high release rate of arginine (75.0 μmol/cm 2 ) at 2 h, and a more sustainable, controlled release rate (up to 0.2 μmol/cm 2 ) at 30 days. Conclusions Incorporation of 7% arginine did not affect the physical and mechanical properties of the adhesive. Arginine was released from the adhesive at a rate and concentration that exhibited antibacterial effects, regardless of shifts in biofilm conditions such as sugar availability and pH. Clinical significance Secondary caries is recognized as the main reason for failure of dental restorations. The development of an arginine-based adhesive system has the potential to dramatically reduce the incidence and severity of secondary caries in adhesive restorations in a very economical fashion.

Novel approaches using OMICS techniques enable a collective assessment of multiple related biological units, including genes, gene expression, proteins, and metabolites. In the past decade, next-generation sequencing (NGS) technologies were improved by longer sequence reads and the development of genome databases and user-friendly pipelines for data analysis, all accessible at lower cost. This has generated an outburst of high-Throughput data. The application of OMICS has provided more depth to existing hypotheses as well as new insights in the etiology of dental caries. For example, the determination of complete bacterial microbiomes of oral samples rather than selected species, together with oral metatranscriptome and metabolome analyses, supports the viewpoint of dysbiosis of the supragingival biofilms. In addition, metabolome studies have been instrumental in disclosing the contributions of major pathways for central carbon and amino acid metabolisms to biofilm pH homeostasis. New, often noncultured, oral streptococci have been identified, and their phenotypic characterization has revealed candidates for probiotic therapy. Although findings from OMICS research have been greatly informative, problems related to study design, data quality, integration, and reproducibility still need to be addressed. Also, the emergence and continuous updates of these computationally demanding technologies require expertise in advanced bioinformatics for reliable interpretation of data. Despite the obstacles cited above, OMICS research is expected to encourage the discovery of novel caries biomarkers and the development of next-generation diagnostics and therapies for caries control. These observations apply equally to the study of other oral diseases.

Approaches for managing carious tissues during cavity preparations vary considerably among clinicians, which may reflect inconsistencies in the teaching of this subject by dental schools. The aims of this study were to investigate practices related to the preclinical and clinical teaching of caries removal at U.S. dental schools and the relationship between that teaching and requirements for U.S. dental licensure examinations. The electronic survey included questions about terminology, methods, instruments and materials, treatment planning, criteria for clinical exams, faculty calibration sessions, and licensure exams. The faculty members at U.S. dental schools responsible for teaching cariology were invited to participate; 54 of the 65 schools had identified a contact person at the time of the survey in October 2015. Of those 54 invited to participate, 43 completed the survey (response rate of 79.6%). Most of the respondents indicated that depth of carious lesions was a clinical determinant of the amount of carious dentin being removed in cavity preparations. Caries removal was used as a criterion in restorative clinical examinations by 95% of responding schools. Marked differences were observed regarding the criteria used for assessment and removal of carious tissues, management of deep carious lesions, and definition of "caries remaining at cavity preparations," which is considered a critical error on licensure exams. Faculty calibration sessions on caries removal were reported to occur in 65% of these schools and at different time frames. Overall, the study found a wide range of teaching practices related to caries removal. Best evidence in caries management needs to be aligned with teaching and the criteria used to calibrate faculty members and examiners.

Health-associated biofilms in the oral cavity are composed of a diverse group of microbial species that can foster an environment that is less favorable for the outgrowth of dental caries pathogens, like Streptococcus mutans. A novel oral bacterium, designated Streptococcus A12, was previously isolated from supragingival dental plaque of a caries-free individual and was shown to interfere potently with the growth and virulence properties of S. mutans. In this study, we applied functional genomics to begin to identify molecular mechanisms used by A12 to antagonize, and to resist the antagonistic factors of, S. mutans. Using bioinformatics, genes that could encode factors that enhance the ability of A12 to compete with S. mutans were identified. Selected genes, designated potential competitive factors (pcf), were deleted. Certain mutant derivatives showed a reduced capacity to compete with S. mutans compared to that of the parental strain. The A12 pcfO mutant lost the ability to inhibit comX-inducing peptide (XIP) signaling by S. mutans, while mutants with changes in the pcfFEG locus were impaired in sensing of, and were more sensitive to, the lantibiotic nisin. Loss of PcfV, annotated as a colicin V biosynthetic protein, resulted in diminished antagonism of S. mutans. Collectively, the data provide new insights into the complexities and variety of factors that affect biofilm ecology and virulence. Continued exploration of the genomic and physiological factors that distinguish commensals from truly beneficial members of the oral microbiota will lead to a better understanding of the microbiome and new approaches to promote oral health. IMPORTANCE Advances in defining the composition of health-associated biofilms have highlighted the important role of beneficial species in maintaining health. Comparatively little, however, has been done to address the genomic and physiological bases underlying the probiotic mechanisms of beneficial commensals. In this study, we explored the ability of a novel oral bacterial isolate, Streptococcus A12, to compete with the dental pathogen Streptococcus mutans using various gene products with diverse functions. A12 displayed enhanced competitiveness by (i) disrupting intercellular communication pathways of S. mutans, (ii) sensing and resisting antimicrobial peptides, and (iii) producing factors involved in the production of a putative antimicrobial compound. Research on the probiotic mechanisms employed by Streptococcus A12 is providing essential insights into how beneficial bacteria may help maintain oral health, which will aid in the development of biomarkers and therapeutics that can improve the practice of clinical dentistry.

Patients with glycogen storage disease (GSD) are either orally fed (ORF) or gastronomy-tube fed (GTF) with cornstarch to maintain normal glucose levels. It is not known whether the use of cornstarch affects the microbiological oral profile of patients with GSD. Thus, the purpose of this study was to compare supragingival and subgingival plaque samples collected from 53 participants with GSD (2-56 years)—29 ORF and 24 GTF. The 16S sequence bacterial profiles of plaque DNA were obtained and a total of 768 probes were detected across the plaque groups. Orally fed patients showed higher means of cariogenic species and periodontal health-associated species, whereas GTF patients showed higher means of periopathogenic species (P < .05). Orally fed patients exhibited high levels of caries pathogens and lower levels of periodontal pathogens possibly due to the acidic environment created by their cornstarch diet, when compared to GTF patients.

The high prevalence of early childhood caries and many general dentists’ reluctance to treat young children and pregnant women demand new educational programs to foster delivery of oral health services. The aim of this study was to evaluate the impact of an Infant Oral Health Program (IOHP) at the University of Florida College of Dentistry on dental students’ knowledge about and willingness to provide dental care for infants, children up to three years of age, and pregnant women. A total of 233 dental students in the first through fourth years and recent graduates completed a survey that assessed the educational outcomes of the IOHP; only the fourth-year students had received IOHP training. The results showed that females were more likely than males to provide counseling to caregivers about dental and physical development (p=0.024) and to offer restorative treatment to young children (p=0.021). Older students were more likely than younger students to provide restorative treatment (p=0.013). A greater percentage of IOHP-trained students (96%) reported knowing how to use the lap examination technique compared with untrained students (71%; p

Streptococcus mutans is an oral bacterium considered to play a major role in the development of dental caries. This study aimed to investigate the prevalence of S. mutans in active and arrested dentine carious lesions of children with early childhood caries and to examine the expression profile of selected S. mutans genes associated with survival and virulence, within the same carious lesions. Dentine samples were collected from 29 active and 16 arrested carious lesions that were diagnosed in preschool children aged 2-5 years. Total RNA was extracted from the dentine samples, and reverse transcription quantitative real-time PCR analyses were performed for the quantification of S. mutans and for analyses of the expression of S. mutans genes associated with bacterial survival (atpD, nox, pdhA) and virulence (fabM and aguD). There was no statistically significant difference in the prevalence of S. mutans between active and arrested carious lesions. Expression of the tested genes was detected in both types of carious dentine. The pdhA (p = 0.04) and aguD (p = 0.05) genes were expressed at higher levels in arrested as compared to active lesions. Our findings revealed that S. mutans is part of the viable microbial community in active and arrested dentine carious lesions. The increase in expression of the pdhA and aguD genes in arrested lesions is likely due to the unfavourable environmental conditions for microbial growth, inherent to this type of lesions.

Representative type strains are commonly used to characterize bacterial species, yet species are phenotypically and genotypically heterogeneous. Conclusions about strain physiology and activity based on a single strain therefore may be inappropriate and misleading. When selecting strains for probiotic use, the assumption that all strains within a species share the same desired probiotic characteristics may result in selection of a strain that lacks the desired traits, and therefore makes a minimally effective or ineffective probiotic. Health-associated oral streptococci are promising candidates for anticaries probiotics, but strains need to be carefully selected based on observed phenotypes. We characterized the genotypes and anticaries phenotypes of strains from 10 species of oral streptococci and demonstrate poor correlation between genotype and phenotype across all species., Health-associated oral Streptococcus species are promising probiotic candidates to protect against dental caries. Ammonia production through the arginine deiminase system (ADS), which can increase the pH of oral biofilms, and direct antagonism of caries-associated bacterial species are desirable properties for oral probiotic strains. ADS and antagonistic activities can vary dramatically among individuals, but the genetic basis for these differences is unknown. We sequenced whole genomes of a diverse set of clinical oral Streptococcus isolates and examined the genetic basis of variability in ADS and antagonistic activities. A total of 113 isolates were included and represented 10 species: Streptococcus australis, A12-like, S. cristatus, S. gordonii, S. intermedius, S. mitis, S. oralis including S. oralis subsp. dentisani, S. parasanguinis, S. salivarius, and S. sanguinis. Mean ADS activity and antagonism on Streptococcus mutans UA159 were measured for each isolate, and each isolate was whole genome shotgun sequenced on an Illumina MiSeq. Phylogenies were built of genes known to be involved in ADS activity and antagonism. Several approaches to correlate the pan-genome with phenotypes were performed. Phylogenies of genes previously identified in ADS activity and antagonism grouped isolates by species, but not by phenotype. A genome-wide association study (GWAS) identified additional genes potentially involved in ADS activity or antagonism across all the isolates we sequenced as well as within several species. Phenotypic heterogeneity in oral streptococci is not necessarily reflected by genotype and is not species specific. Probiotic strains must be carefully selected based on characterization of each strain and not based on inclusion within a certain species. IMPORTANCE Representative type strains are commonly used to characterize bacterial species, yet species are phenotypically and genotypically heterogeneous. Conclusions about strain physiology and activity based on a single strain therefore may be inappropriate and misleading. When selecting strains for probiotic use, the assumption that all strains within a species share the same desired probiotic characteristics may result in selection of a strain that lacks the desired traits, and therefore makes a minimally effective or ineffective probiotic. Health-associated oral streptococci are promising candidates for anticaries probiotics, but strains need to be carefully selected based on observed phenotypes. We characterized the genotypes and anticaries phenotypes of strains from 10 species of oral streptococci and demonstrate poor correlation between genotype and phenotype across all species.

An expert panel convened by the American Dental Association Council on Scientific Affairs and the Center for Evidence-Based Dentistry conducted a systematic review and formulated evidence-based clinical recommendations for the arrest or reversal of noncavitated and cavitated dental caries using nonrestorative treatments in children and adults.The authors conducted a systematic search of the literature in MEDLINE and Embase via Ovid, Cochrane CENTRAL, and Cochrane database of systematic reviews to identify randomized controlled trials reporting on nonrestorative treatments for noncavitated and cavitated carious lesions. The authors used the Grading of Recommendations Assessment, Development and Evaluation approach to assess the certainty in the evidence and move from the evidence to the decisions.The expert panel formulated 11 clinical recommendations, each specific to lesion type, tooth surface, and dentition. Of the most effective interventions, the panel provided recommendations for the use of 38% silver diamine fluoride, sealants, 5% sodium fluoride varnish, 1.23% acidulated phosphate fluoride gel, and 5,000 parts per million fluoride (1.1% sodium fluoride) toothpaste or gel, among others. The panel also provided a recommendation against the use of 10% casein phosphopeptide-amorphous calcium phosphate.Although the recommended interventions are often used for caries prevention, or in conjunction with restorative treatment options, these approaches have shown to be effective in arresting or reversing carious lesions. Clinicians are encouraged to prioritize use of these interventions based on effectiveness, safety, and feasibility.

The aims of this study were to describe the processes used to train dental faculty members in case-based learning (CBL) and to determine their beliefs about the anticipated implementation of CBL and perceptions of actual implementation following use of the CBL approach. Participants were dental faculty members at the University of Florida who received a four-day intensive training course in the use of CBL. Two focus groups were conducted. The first occurred during training to assess how the participants anticipated using CBL. The second was conducted during the faculty members' implementation of CBL. All 19 trainees participated in focus group 1 (100%). During the course of the study, two faculty members left the school; of the remaining 17, 12 participated in focus group 2 (participation rate of 71%). The findings showed that initially the faculty members were hesitant and uncertain about using CBL. Following implementation, those issues dissipated, as the participants began to consider how to optimize the effectiveness of CBL as a legitimate method for fostering student ownership of learning and active participation. Understanding what CBL means for individual educators at varying stages of change will likely allow the dental education community to better anticipate and address tensions and challenges that faculty members are likely to experience.

The oral microbiota associated with the initiation and progression of dental caries has yet to be fully characterized. The Human Oral Microbe Identification Using Next-Generation Sequencing (HOMI NGS ) approach was used to analyze the microbiomes of site-specific supragingival dental plaques from children with different caries status. Fifty-five children (2 to 7 years of age) were assessed at baseline and at 12 months and grouped as caries free (CF), caries active with enamel lesions (CAE), and caries active with dentin carious lesions (CA). Plaque samples from caries-free tooth surfaces (PF) and from enamel carious lesions (PE) and dentin carious lesions (PD) were collected. 16S community profiles were obtained by HOMI NGS , and 408 bacterial species and 84 genus probes were assigned. Plaque bacterial communities showed temporal stability, as there was no significant difference in beta diversity values between the baseline and 12-month samples. Irrespective of collection time points, the microbiomes of healthy tooth surfaces differed substantially from those found during caries activity. All pairwise comparisons of beta diversity values between groups were significantly different ( P < 0.05), except for comparisons between the CA-PF, CAE-PE, and CA-PE groups. Streptococcus genus probe 4 and Neisseria genus probe 2 were the most frequently detected taxa across the plaque groups, followed by Streptococcus sanguinis , which was highly abundant in CF-PF. Well-known acidogenic/aciduric species such as Streptococcus mutans , Scardovia wiggsiae , Parascardovia denticolens , and Lactobacillus salivarius were found almost exclusively in CA-PD. The microbiomes of supragingival dental plaque differ substantially among tooth surfaces and children of different caries activities. In support of the ecological nature of caries etiology, a steady transition in community species composition was observed with disease progression.

Knowledge acquired from microbiological studies of oral arginine metabolism over the past 40 years has recently found its way to clinical application in dentistry. Novel arginine formulations incorporated into dentifrices have proven to be effective as an approach to prevent dental caries. In oral biofilms, arginine metabolism via the arginine deiminase system (ADS) produces ammonia, which inhibits tooth demineralization by neutralizing glycolytic acids and by suppressing the emergence of a cariogenic microflora. Evidence from translational science studies suggests that arginine metabolism in oral biofilms decreases the risk for caries development. These studies have expanded the focus of caries research beyond cariogenic bacteria to health-related, ADS-positive bacteria. The focus of this article is on the contribution of arginine metabolism to pH homeostasis in oral biofilms and its impact on the etiology of dental caries.

Introduction As educators, we strive to evaluate the work of our students as objectively as possible. Assessments are used to provide feedback that will encourage and motivate students to continually seek evidence-based knowledge and understanding while improving their capacity to apply critical thinking skills in clinical situations. We have come to understand that many students focus on earning the highest attainable grade rather than concentrating on what they need to learn. In our exploration of this, it became apparent that we could use methods of assessment to motivate the student to learn. Clinical assessment of dental students involves a thorough evaluation of both quality of performance and breadth of experience. This resource focuses on the quality assessment of clinical patient care. Methods Included in this resource is a rubric for daily clinical formative assessments, semi-independent formative skills assessments for the junior-level student, and independent summative skills assessments/competency evaluations for the senior-level student. The Level I assessment is used both to evaluate a student's psychomotor ability and their ability to select appropriate cases, manage patients and make evidence-based decisions while providing dental treatment with limited faculty intervention. Level II assessments evaluate the same in addition to the student's ability to operate independently while providing dental treatment without faculty intervention. Results The operative dentistry clinical assessments in this resource are designed for second, third, and fourth-year dental students. It has been used with approximately 250 students as an overall evaluation of clinical performance by semester. Discussion The clinical assessment of students has changed form many times over the years. With the addition of a competency-based curriculum years ago, the feedback given to students was decreased significantly in our school. Semester grades were primarily based on successful completion of the competency-based assessment. The assessment system described here provides tools for continual, calibrated feedback to students which promotes self-learning.

The ability of certain oral biofilm bacteria to moderate pH through arginine metabolism by the arginine deiminase system (ADS) is a deterrent to the development of dental caries. Here, we characterize a novel Streptococcus strain, designated strain A12, isolated from supragingival dental plaque of a caries-free individual. A12 not only expressed the ADS pathway at high levels under a variety of conditions but also effectively inhibited growth and two intercellular signaling pathways of the dental caries pathogen Streptococcus mutans . A12 produced copious amounts of H 2 O 2 via the pyruvate oxidase enzyme that were sufficient to arrest the growth of S. mutans . A12 also produced a protease similar to challisin (Sgc) of Streptococcus gordonii that was able to block the competence-stimulating peptide (CSP)–ComDE signaling system, which is essential for bacteriocin production by S. mutans . Wild-type A12, but not an sgc mutant derivative, could protect the sensitive indicator strain Streptococcus sanguinis SK150 from killing by the bacteriocins of S. mutans . A12, but not S. gordonii , could also block the XIP ( com X - i nducing p eptide) signaling pathway, which is the proximal regulator of genetic competence in S. mutans , but Sgc was not required for this activity. The complete genome sequence of A12 was determined, and phylogenomic analyses compared A12 to streptococcal reference genomes. A12 was most similar to Streptococcus australis and Streptococcus parasanguinis but sufficiently different that it may represent a new species. A12-like organisms may play crucial roles in the promotion of stable, health-associated oral biofilm communities by moderating plaque pH and interfering with the growth and virulence of caries pathogens.

Streptococcus mutans is an oral bacterium considered to play a major role in the development of dental caries. This study aimed to investigate the prevalence of S. mutans in active and arrested dentine carious lesions of children with early childhood caries and to examine the expression profile of selected S. mutans genes associated with survival and virulence, within the same carious lesions. Dentine samples were collected from 29 active and 16 arrested carious lesions that were diagnosed in preschool children aged 2-5 years. Total RNA was extracted from the dentine samples, and reverse transcription quantitative real-time PCR analyses were performed for the quantification of S. mutans and for analyses of the expression of S. mutans genes associated with bacterial survival (atpD, nox, pdhA) and virulence (fabM and aguD). There was no statistically significant difference in the prevalence of S. mutans between active and arrested carious lesions. Expression of the tested genes was detected in both types of carious dentine. The pdhA (p = 0.04) and aguD (p = 0.05) genes were expressed at higher levels in arrested as compared to active lesions. Our findings revealed that S. mutans is part of the viable microbial community in active and arrested dentine carious lesions. The increase in expression of the pdhA and aguD genes in arrested lesions is likely due to the unfavourable environmental conditions for microbial growth, inherent to this type of lesions.

Alkali production by oral bacteria is believed to have a major impact on oral microbial ecology and to be inhibitory to the initiation and progression of dental caries. A substantial body of evidence is beginning to accumulate that indicates the modulation of the alkalinogenic potential of dental biofilms may be a promising strategy for caries control. This brief review highlights recent progress toward understanding molecular genetic and physiologic aspects of important alkali-generating pathways in oral bacteria, and the role of alkali production in the ecology of dental biofilms in health and disease.

The authors conducted a study to quantify the reasons for restoring noncarious tooth defects (NCTDs) by dentists in The Dental Practice-Based Research Network (DPBRN) and to assess the tooth, patient and dentist characteristics associated with those reasons.Data were collected by 178 DPBRN dentists regarding the placement of 1,301 consecutive restorations owing to NCTDs. Information gathered included the main clinical reason, other than dental caries, for restoration of previously unrestored permanent tooth surfaces; characteristics of patients who received treatment; dentists' and dental practices' characteristics; teeth and surfaces restored; and restorative materials used.Dentists most often placed restorations to treat lesions caused by abrasion, abfraction or erosion (AAE) (46 percent) and tooth fracture (31 percent). Patients 41 years or older received restorations mainly because of AAE (P.001). Premolars and anterior teeth were restored mostly owing to AAE; molars were restored mostly owing to tooth fracture (P.001). Dentists used directly placed resin-based composite (RBC) largely to restore AAE lesions and fractured teeth (P.001).Among DPBRN practices, AAE and tooth fracture were the main reasons for restoring noncarious tooth surfaces. Pre-molars and anterior teeth of patients 41 years and older are most likely to receive restorations owing to AAE; molars are most likely to receive restorations owing to tooth fracture. Dentists restored both types of NCTDs most often with RBC.

Background An expert panel convened by the American Dental Association Council on Scientific Affairs and the Center for Evidence-Based Dentistry conducted a systematic review and formulated evidence-based clinical recommendations for the arrest or reversal of noncavitated and cavitated dental caries using nonrestorative treatments in children and adults. Types of Studies Reviewed The authors conducted a systematic search of the literature in MEDLINE and Embase via Ovid, Cochrane CENTRAL, and Cochrane database of systematic reviews to identify randomized controlled trials reporting on nonrestorative treatments for noncavitated and cavitated carious lesions. The authors used the Grading of Recommendations Assessment, Development and Evaluation approach to assess the certainty in the evidence and move from the evidence to the decisions. Results The expert panel formulated 11 clinical recommendations, each specific to lesion type, tooth surface, and dentition. Of the most effective interventions, the panel provided recommendations for the use of 38% silver diamine fluoride, sealants, 5% sodium fluoride varnish, 1.23% acidulated phosphate fluoride gel, and 5,000 parts per million fluoride (1.1% sodium fluoride) toothpaste or gel, among others. The panel also provided a recommendation against the use of 10% casein phosphopeptide–amorphous calcium phosphate. Conclusions and Practical Implications Although the recommended interventions are often used for caries prevention, or in conjunction with restorative treatment options, these approaches have shown to be effective in arresting or reversing carious lesions. Clinicians are encouraged to prioritize use of these interventions based on effectiveness, safety, and feasibility.

This study investigated the concordance between pre- and postoperative assessments of primary caries lesion depths by dentists from The Dental Practice-Based Research Network (DPBRN; www.DentalPBRN.org). A total of 229 DPBRN dentists collected data on 8,351 consecutive restorations inserted due to primary caries in 5,810 patients. Dentists estimated the preoperative depth of caries lesions based on the diagnostic methods they typically used. The preoperative depth was then compared to the postoperative depth, which dentists determined using actual clinical observation. Both estimated and observed depths were recorded as being in the outer half (E1) or inner half (E2) of enamel, or in the outer third (D1), middle third (D2) or inner third (D3) of dentin. Most restorations were placed to treat lesions that were preoperatively assessed as extending to the D1 (53%) and D2 (25%) depths. Of the restored caries lesions, 10% were preoperatively assessed as being limited to E2 depth and 3% to E1 depth. The majority of the restored enamel lesions were located on occlusal surfaces. Preoperative estimates of caries lesion depth were more concordant with postoperative depths when the lesion was at an advanced stage: 88% concordance at the D3 depth, compared to 54% concordance at the E1 depth. DPBRN dentists can discriminate caries lesions at different depths, but the accuracy of their depth assessments was higher for dentin than for enamel lesions. In general, DPBRN dentists were more likely to underestimate than overestimate the depth of caries lesions, and the extent of underestimation was greater for enamel than for dentin lesions.

Objective The authors conducted a study to identify and quantify the reasons used by dentists in The Dental Practice-Based Research Network (DPBRN) for placing restorations on unrestored permanent tooth surfaces and the dental materials they used in doing so. Methods A total of 229 DPBRN practitioner-investigators provided data from their practices regarding 9,890 consecutive restorations in 5,810 patients. Information the practitioner-investigators provided included their reasons for restoring the teeth, the specific teeth and surfaces they restored and the restorative materials they used. Results Primary caries (85 percent of teeth, 8,351 of 9,890) and noncarious defects (15 percent, 1,479 of 9,890) were the main reasons participants gave for placing restorations. Participants placed restorations necessitated by caries most frequently on occlusal surfaces (49 percent, 4,091 of 8,351). They used amalgam for 47 percent of the molar restorations and 45 percent of the premolar restorations. They used directly placed resin-based composite (RBC) for 48 percent of the molar restorations, 50 percent of the premolar restorations and 93 percent of the anterior restorations. Conclusion DPBRN practitioner-investigators cited dental caries on occlusal and proximal surfaces of molar teeth as the main reasons for placing restorations on previously unrestored tooth surfaces. RBC was the material they used most commonly for occlusal and anterior restorations. Amalgam remains the material of choice to restore posterior teeth with proximal caries, although the authors noted significant differences in the use of amalgam and RBC by dentists in various regions of the DPBRN.

The RelA, RelP, and RelQ enzymes are responsible for the production of the alarmone (p)ppGpp in Streptococcus mutans . A strain lacking all three synthetases (Δ relAPQ ) does not grow in minimal medium lacking the branched-chain amino acids (BCAA) leucine or valine but grows well if isoleucine is also omitted. Here, we investigated whether there was a correlation between growth in the absence of leucine and valine with (p)ppGpp pools and the activation of CodY. By using a combination of single, double, and triple mutants lacking the (p)ppGpp synthetase enzymes, we demonstrated that the ability to grow in the absence of leucine or valine required basal levels of (p)ppGpp production by RelP and RelQ. The introduction of a codY mutation into the Δ relAPQ strain fully restored growth in medium lacking leucine or valine, revealing that the growth-defective phenotype of Δ relAPQ was directly linked to CodY. Lowering GTP levels through the addition of decoyinine did not alleviate CodY repression or affect the expression of genes involved in BCAA biosynthesis, suggesting that S. mutans CodY is not activated by GTP. The results of phenotypic studies revealed that the codY mutant had a reduced capacity to form biofilms and that its growth was more sensitive to low pH, showing a role for CodY in two key virulence properties of S. mutans . Microarray results revealed the extent of the CodY regulon. Notably, the identification of putative CodY-binding boxes upstream of genes that were downregulated in the codY mutant indicates that CodY may also function as a transcriptional activator in S. mutans .

CcpA globally regulates transcription in response to carbohydrate availability in many gram-positive bacteria, but its role in Streptococcus mutans remains enigmatic. Using the fructan hydrolase ( fruA ) gene of S. mutans as a model, we demonstrated that CcpA plays a direct role in carbon catabolite repression (CCR). Subsequently, the expression of 170 genes was shown to be differently expressed (≥2-fold) in glucose-grown wild-type (UA159) and CcpA-deficient (TW1) strains ( P ≤ 0.001). However, there were differences in expression of only 96 genes between UA159 and TW1 when cells were cultivated with the poorly repressing substrate galactose. Interestingly, 90 genes were expressed differently in wild-type S. mutans when glucose- and galactose-grown cells were compared, but the expression of 515 genes was altered in the CcpA-deficient strain in a similar comparison. Overall, our results supported the hypothesis that CcpA has a major role in CCR and regulation of gene expression but revealed that in S. mutans there is a substantial CcpA-independent network that regulates gene expression in response to the carbohydrate source. Based on the genetic studies, biochemical and physiological experiments demonstrated that loss of CcpA impacts the ability of S. mutans to transport and grow on selected sugars. Also, the CcpA-deficient strain displayed an enhanced capacity to produce acid from intracellular stores of polysaccharides, could grow faster at pH 5.5, and could acidify the environment more rapidly and to a greater extent than the parental strain. Thus, CcpA directly modulates the pathogenic potential of S. mutans through global control of gene expression.

The current dogma implicating RelA as the sole enzyme controlling (p)ppGpp production and degradation in Gram-positive bacteria does not apply to Streptococcus mutans. We have now identified and characterized two genes, designated as relP and relQ, encoding novel enzymes that are directly involved in (p)ppGpp synthesis. Additionally, relP is co-transcribed with a two-component signal transduction system (TCS). Analysis of the (p)ppGpp synthetic capacity of various mutants and the behaviour of strains lacking combinations of the synthetase enzymes have revealed a complex regulon and fundamental differences in the way S. mutans manages alarmone production compared with bacterial paradigms. The functionality of the RelP and RelQ enzymes was further confirmed by demonstrating that expression of relP and relQ restored growth of a (p)ppGpp(0) Escherichia coli strain in minimal medium, SMG and on medium containing 3-amino-1,2,4-triazole, and by demonstrating (p)ppGpp production in various complemented mutant strains of E. coli and S. mutans. Notably, RelQ, and RelP and the associated TCS, are harboured in some, but not all, pathogenic streptococci and related Gram-positive organisms, opening a new avenue to explore the variety of strategies employed by human and animal pathogens to survive in adverse conditions that are peculiar to environments in their hosts.

The high prevalence of early childhood caries and many general dentists' reluctance to treat young children and pregnant women demand new educational programs to foster delivery of oral health services. The aim of this study was to evaluate the impact of an Infant Oral Health Program (IOHP) at the University of Florida College of Dentistry on dental students' knowledge about and willingness to provide dental care for infants, children up to three years of age, and pregnant women. A total of 233 dental students in the first through fourth years and recent graduates completed a survey that assessed the educational outcomes of the IOHP; only the fourth-year students had received IOHP training. The results showed that females were more likely than males to provide counseling to caregivers about dental and physical development (p=0.024) and to offer restorative treatment to young children (p=0.021). Older students were more likely than younger students to provide restorative treatment (p=0.013). A greater percentage of IOHP-trained students (96%) reported knowing how to use the lap examination technique compared with untrained students (71%; p0.001). IOHP-trained students were the most uncomfortable dealing with a crying child in the dental chair (p=0.0175). The graduates and fourth-year students were significantly more likely than the other cohorts to provide preventive (p=0.001) and restorative (p0.001) care for pregnant women. The graduates were least likely to use some form of caries risk assessment (p0.001). These findings highlight the need for earlier and greater exposure to the IOHP and the importance of promoting awareness about risk assessment and oral disease management.

The aims of this study were to describe the processes used to train dental faculty members in case-based learning (CBL) and to determine their beliefs about the anticipated implementation of CBL and perceptions of actual implementation following use of the CBL approach. Participants were dental faculty members at the University of Florida who received a four-day intensive training course in the use of CBL. Two focus groups were conducted. The first occurred during training to assess how the participants anticipated using CBL. The second was conducted during the faculty members' implementation of CBL. All 19 trainees participated in focus group 1 (100%). During the course of the study, two faculty members left the school; of the remaining 17, 12 participated in focus group 2 (participation rate of 71%). The findings showed that initially the faculty members were hesitant and uncertain about using CBL. Following implementation, those issues dissipated, as the participants began to consider how to optimize the effectiveness of CBL as a legitimate method for fostering student ownership of learning and active participation. Understanding what CBL means for individual educators at varying stages of change will likely allow the dental education community to better anticipate and address tensions and challenges that faculty members are likely to experience.

The aim of the present study was to evaluate the colonization profile and clonal distribution of Streptococcus mutans isolated from oral cavities that presented coronal and root caries lesions. The isolation and biochemical identification of mutans streptococci were carried out by using saliva samples, dental plaque, and tissue from the caries lesions. In order to confirm their molecular identity, S. mutans and Streptococcus sobrinus were submitted to the PCR method, using specific primers for portions of the glucosyltransferase genes (gtfB and gtfI, respectively). The AP-PCR method was used to detect the genetic polymorphism of S. mutans strains. Among the isolated and identified species, S. mutans showed a significantly greater frequency of isolation (59.2%) than the other mutans streptococci. Each of the subjects harbored two to ten genotypes of S. mutans, randomly distributed in different sites. S. mutans genotypes showed no evidence of variability in colonizing noncarious and carious surfaces within the same individual, nor evidence of etiologic differences between coronal and root caries. This study showed that no particular genotype of S. mutans is uniquely associated with the initiation and progression of caries, and that root and coronal caries can emerge in the presence of a broad spectrum of S. mutans clones.