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ABSTRACTArginine catabolism by the bacterial arginine deiminase system (ADS) has anticariogenic properties through the production of ammonia, which modulates the pH of the oral environment. Given the potential protective capacity of the ADS pathway, the exploitation of ADS-competent oral microbes through pre- or probiotic applications is a promising therapeutic target to prevent tooth decay. To date, most investigations of the ADS in the oral cavity and its relation to caries have focused on indirect measures of activity or on specific bacterial groups, yet the pervasiveness and rate of expression of the ADS operon in diverse mixed microbial communities in oral health and disease remain an open question. Here, we use a multivariate approach, combining ultra-deep metatranscriptomic sequencing with paired metataxonomic and in vitro citrulline quantification to characterize the microbial community and ADS operon expression in healthy and late-stage cavitated teeth. While ADS activity is higher in healthy teeth, we identify multiple bacterial lineages with upregulated ADS activity on cavitated teeth that are distinct from those found on healthy teeth using both reference-based mapping and de novo assembly methods. Our dual metataxonomic and metatranscriptomic approach demonstrates the importance of species abundance for gene expression data interpretation and that patterns of differential expression can be skewed by low-abundance groups. Finally, we identify several potential candidate probiotic bacterial lineages within species that may be useful therapeutic targets for the prevention of tooth decay and propose that the development of a strain-specific, mixed-microbial probiotic may be a beneficial approach given the heterogeneity of taxa identified here across health groups.IMPORTANCETooth decay is the most common preventable chronic disease, affecting more than two billion people globally. The development of caries on teeth is primarily a consequence of acid production by cariogenic bacteria that inhabit the plaque microbiome. Other bacterial strains in the oral cavity may suppress or prevent tooth decay by producing ammonia as a byproduct of the arginine deiminase metabolic pathway, increasing the pH of the plaque biofilm. While the benefits of arginine metabolism on oral health have been extensively documented in specific bacterial groups, the prevalence and consistency of arginine deiminase system (ADS) activity among oral bacteria in a community context remain an open question. In the current study, we use a multi-omics approach to document the pervasiveness of the expression of the ADS operon in both health and disease to better understand the conditions in which ADS activity may prevent tooth decay.

Abstract Background Subgingival microbiome in disease-associated subgingival sites is known to be dysbiotic and significantly altered. In patients with rheumatoid arthritis (RA), the extent of dysbiosis in disease- and health-associated subgingival sites is not clear. Methods 8 RA and 10 non-RA subjects were recruited for this pilot study. All subjects received full oral examination and underwent collection of subgingival plaque samples from both shallow (periodontal health-associated, probing depth ≤ 3mm) and deep subgingival sites (periodontal disease-associated, probing depth ≥ 4 mm). RA subjects also had rheumatological evaluation. Plaque community profiles were analyzed using 16 S rRNA sequencing. Results The phylogenetic diversity of microbial communities in both RA and non-RA controls was significantly higher in deep subgingival sites compared to shallow sites (p = 0.022), and the overall subgingival microbiome clustered primarily according to probing depth (i.e. shallow versus deep sites), and not separated by RA status. While a large number of differentially abundant taxa and gene functions was observed between deep and shallow sites as expected in non-RA controls, we found very few differentially abundant taxa and gene functions between deep and shallow sites in RA subjects. In addition, compared to non-RA controls, the UniFrac distances between deep and shallow sites in RA subjects were smaller, suggesting increased similarity between deep and shallow subgingival microbiome in RA. Streptococcus parasanguinis and Actinomyces meyeri were overabundant in RA subjects, while Gemella morbillorum, Kingella denitrificans, Prevotella melaninogenica and Leptotrichia spp. were more abundant in non-RA subjects. Conclusions The aggregate subgingival microbiome was not significantly different between individuals with and without rheumatoid arthritis. Although the differences in the overall subgingival microbiome was driven primarily by probing depth, in contrast to the substantial microbiome differences typically seen between deep and shallow sites in non-RA patients, the microbiome of deep and shallow sites in RA patients were more similar to each other. These results suggest that factors associated with RA may modulate the ecology of subgingival microbiome and its relationship to periodontal disease, the basis of which remains unknown but warrants further investigation.

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,

ABSTRACT 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.

Background: Parental reports of their children’s health status is integral to pediatric medical and dental care. Therefore, understanding the accuracy of such reports is vital. Our objectives were to (1) assess the correlation between maternal reports of their children’s indicators of caries experience (subjective assessment) and actual caries status determined by oral examination (objective assessment), and (2) identify potential modifiers of this correlation. Methods: Longitudinal data from the Avon longitudinal study of parents and children (n = 1429) was used to assess the correlation between maternal reports of the number of missing and filled teeth of children aged 38 months, 54 months, and 5.5 years and clinical oral examinations of decayed, missing and filled teeth conducted when the same children were 31, 43 months, and five years of age. Homogeneity chi-square tests assessed differences in correlations according to sociodemographic factors. Results: Overall, we found a statistically significant correlation that was weak to moderate in magnitude. Maternal reports of missing teeth at 38 months was significantly correlated with decayed teeth, 0.27 (p < 0.001); missing teeth, 0.23 (p < 0.001), and the decayed, missing and filled (dmft) index, 0.35 (p < 0.001) based on oral examination at 31 months. A maternal report of filled teeth at 54 months was significantly correlated with decayed teeth, 0.30 (p < 0.001); filled teeth 0.30 (p < 0.001), and dmft 0.40 (p < 0.001) at 43 months. Mothers tended to underestimate the extent of missing and filled teeth in their children irrespective of the child’s age, but the extent of underestimation was greater among younger children. Maternal age, education level, and whether the child had ever visited a dentist were significant modifiers of subjective and objective caries assessments. Conclusions: From a clinical and dental public health perspective, our findings of a weak to moderate correlation of maternal assessments of their children’s caries experience may be concerning when reporting the burden of dental diseases in large population studies or for surveillance purposes that rely on self-reported measures and must therefore be utilized with caution because of the potential to result in underestimated disease burden.

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.

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.

Background: An expert panel convened by the American Dental Association (ADA) Council on Scientific Affairs together with the ADA Science and Research Institute's program for Clinical and Translational Research conducted a systematic review and developed recommendations for the treatment of moderate and advanced cavitated caries lesions in patients with vital, nonendodontically treated primary and permanent teeth., Types of Studies Reviewed: The authors searched for systematic reviews comparing carious tissue removal (CTR) approaches in Ovid MEDLINE, Embase, Cochrane Database of Systematic Reviews, and Trip Medical Database. The authors also conducted a systematic search for randomized controlled trials comparing direct restorative materials in Ovid MEDLINE, Embase, Cochrane Central Register of Controlled Trials, ClinicalTrials.gov, and the World Health Organization International Clinical Trials Registry Platform. The authors used the Grading of Recommendations Assessment, Development, and Evaluation approach to assess the certainty of the evidence and formulate recommendations., Results: The panel formulated 16 recommendations and good practice statements: 4 on CTR approaches specific to lesion depth and 12 on direct restorative materials specific to tooth location and surfaces involved. The panel conditionally recommended for the use of conservative CTR approaches, especially for advanced lesions. Although the panel conditionally recommended for the use of all direct restorative materials, they prioritized some materials over the use of others for certain clinical scenarios., Practical Implications: The evidence suggests that more conservative CTR approaches may decrease the risk of adverse effects. All included direct restorative materials may be effective in treating moderate and advanced caries lesions on vital, nonendodontically treated primary and permanent teeth., (Copyright © 2023 American Dental Association. Published by Elsevier Inc. All rights reserved.)

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.

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.