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International audience; BACKGROUND: Exceptional episodes of exposure to high levels of persistent organic pollutants have already been associated with developmental defects of enamel among children, but knowledge is still scarce concerning the contribution of background levels of environmental contamination. METHODS: Children of the French PELAGIE mother-child cohort were followed from birth, with collection of medical data and cord blood samples that were used to measure polychlorinated biphenyls (PCBs), organochlorine pesticides (OCs), and perfluorinated alkyl substances (PFASs). At 12 years of age, molar-incisor hypomineralization (MIH) and other enamel defects (EDs) were recorded for 498 children. Associations were studied using logistic regression models adjusted for potential prenatal confounders. RESULTS: An increasing log-concentration of β-HCH was associated with a reduced risk of MIH and EDs (OR = 0.55; 95% CI, 0.32-0.95, and OR = 0.65; 95% CI, 0.43-0.98, respectively). Among girls, intermediate levels of p,p’-DDE were associated with a reduced risk of MIH. Among boys, we observed an increased risk of EDs in association with intermediate levels of PCB 138, PCB 153, PCB 187, and an increased risk of MIH with intermediate levels of PFOA and PFOS. CONCLUSIONS: Two OCs were associated with a reduced risk of dental defects, whereas the associations between PCBs and PFASs and EDs or MIH were generally close to null or sex-specific, with an increased risk of dental defects in boys. These results suggest that POPs could impact amelogenesis. Replication of this study is required and the possible underlying mechanisms need to be explored.

International audience; Despite clinical evidence of poor oral health and hygiene in Parkinson’s disease (PD) patients, the mouth is often overlooked by both patients and the medical community, who generally focus on motor or psychiatric disorders considered more burdensome. Yet, oral health is in a two-way relationship with overall health-a weakened status triggering a decline in the quality of life. Here, we aim at giving a comprehensive overview of oral health disorders in PD, while identifying their etiologies and consequences. The physical (abnormal posture, muscle tone, tremor, and dyskinesia), behavioral (cognitive and neuropsychiatric disorders), and iatrogenic patterns associated with PD have an overall detrimental effect on patients’ oral health, putting them at risk for other disorders (infections, aspiration, pain, malnutrition), reducing their quality of life and increasing their isolation (anxiety, depression, communication issues). Interdisciplinary cooperation for prevention, management and follow-up strategies need to be implemented at an early stage to maintain and improve patients’ overall comfort and condition. Recommendations for practice, including (non-)pharmacological management strategies are discussed, with an emphasis on the neurologists’ role. Of interest, the oral cavity may become a valuable tool for diagnosis and prognosis in the near future (biomarkers). This overlooked but critical issue requires further attention and interdisciplinary research.

International audience; OBJECTIVES:Rapid development of digital technologies and 3D printing provide new tools for orthodontic indirect bonding. The purpose of this in vitro study is to evaluate the clinical acceptability of hard CAD/CAM indirect bonding tray.MATERIAL AND METHODS:Ten soft silicone transfer trays and ten hard CAD/CAM trays were produced, and 200 brackets were placed on them. The brackets were then transferred to twenty stereolithography -printed models by indirect bonding. These models were scanned and digitally compared with the reference model by three-dimensional superimpositions (GOM software). The linear and angular measurements were collected and analyzed.RESULTS:For the CAD/CAM trays, 100% of the mesiodistal, vertical, and transverse measurements of incisors were within the clinically acceptable range of the American Board of Orthodontists (ABO) standards. More specifically, the clinically acceptable linear measurements were between 97 and 100% for silicone trays while they were between 89 and 100% for CAD/CAM trays. The clinically acceptable angular measurements varied between 87 and 100% for the silicone trays and between 79 and 100% for the CAD/CAM trays. Silicone trays were more precise than CAD/CAM trays. The difference was significant for all linear and angular measurements.CONCLUSIONS:While the CAD/CAM group shows clinically acceptable results according to the ABO, silicone remains to be more precise than CAD/CAM for transfer trays and is therefore still the reference.CLINICAL RELEVANCE:We demonstrate here that the orthodontic indirect bondings, whether they are realized using silicone transfer trays or CAD/CAM trays, are clinically acceptable in terms of the repositioning accuracy of brackets.

International audience; The effect of probiotics in improving or maintaining oral health in orthodontic patients is understudied. The aim of this study is to evaluate the effect of probiotic administration in addition to tooth brushing on clinical gingival inflammation, plaque formation, subgingival microbiota composition, and salivary biomarkers of inflammation in adolescents with fixed orthodontic appliances. The present study is a 6-month, double-blind, two-arm, placebo-controlled, single-center trial, in which 116 adolescent volunteers aged 12–16 years will be recruited from the patients of the orthodontics clinic of the University Hospital of Lille, France. Subjects who meet the eligibility criteria will be allocated to one of the following groups: (i) control: two placebo lozenges per day for 90 days together with regular oral hygiene, (ii) test: two probiotic lozenges per day for 90 days together with regular oral hygiene. Clinical assessment and biological sample collection will be performed at baseline, 3 and 6 months. In addition, compliance outcomes and adverse events will be monitored. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

International audience; OBJECTIVE/BACKGROUND: This retrospective non-randomized controlled cohort study aimed to evaluate the efficiency of simultaneous maxillary expansion and mandibular advancement for the management of pediatric OSAS. PATIENTS/METHODS: The sample was composed of 94 children treated with an innovative orthopedic device to correct a Class II malocclusion associated with an OSAS. Polysomnographic recordings were performed before and after the treatment. We also included a group of 113 age-matched control patients who had the same pathologies, but who did not receive the orthopedic treatment at the time they undergone polysomnographic exams. Statistical tests evaluated the significance of the evolution of these data, both in treated and untreated control patients. RESULTS: After nine months (±3 months) of treatment, respiratory OSAS symptoms significantly improved: the AHI significantly decreased as it became inferior to the pathological threshold (

International audience; The complexity of the oral microbiome continues to astound researchers even with the advancement of multi-disciplinary strategies being used to study these microorganisms in relation to the human body. There is extensive literature available that explains how oral bacterial communities exist within the biofilm and maintains a balance with the host immune system, but when this balance is tipped disease can occur. The purpose of this review is to highlight the subgingival microbial compositions during health and periodontal disease using next generation sequencing techniques, as well as determining the types of functional activities that partake during these states. The subgingival microbiota is a fluid structure that can adapt accordingly to the environment and the identification of signature biomarkers may aid in the assessment of risk and disease severity in an individual to complement clinical diagnosis in the future.

International audience; Porphyromonas gingivalis, a Gram-negative anaerobic bacillus present in periodontal disease, is considered one of the major pathogens in periodontitis. A literature search for English original studies, case series and review articles published up to December 2019 was performed using the MEDLINE, PubMed and GoogleScholar databases, with the search terms "Porphyromonas gingivalis" AND the potentially associated condition or systemic disease Abstracts and full text articles were used to make a review of published research literature on P. gingivalis outside the oral cavity. The main points of interest of this narrative review were: (i) a potential direct action of the bacterium and not the systemic effects of the inflammatory acute-phase response induced by the periodontitis, (ii) the presence of the bacterium (viable or not) in the organ, or (iii) the presence of its virulence factors. Virulence factors (gingipains, capsule, fimbriae, hemagglutinins, lipopolysaccharide, hemolysin, iron uptake transporters, toxic outer membrane blebs/vesicles, and DNA) associated with P. gingivalis can deregulate certain functions in humans, particularly host immune systems, and cause various local and systemic pathologies. The most recent studies linking P. gingivalis to systemic diseases were discussed, remembering particularly the molecular mechanisms involved in different infections, including cerebral, cardiovascular, pulmonary, bone, digestive and peri-natal infections. Recent involvement of P. gingivalis in neurological diseases has been demonstrated. P. gingivalis modulates cellular homeostasis and increases markers of inflammation. It is also a factor in the oxidative stress involved in beta-amyloid production.

The effect of probiotics in improving or maintaining oral health in orthodontic patients is understudied. The aim of this study is to evaluate the effect of probiotic administration in addition to tooth brushing on clinical gingival inflammation, plaque formation, subgingival microbiota composition, and salivary biomarkers of inflammation in adolescents with fixed orthodontic appliances. The present study is a 6-month, double-blind, two-arm, placebo-controlled, single-center trial, in which 116 adolescent volunteers aged 12-16 years will be recruited from the patients of the orthodontics clinic of the University Hospital of Lille, France. Subjects who meet the eligibility criteria will be allocated to one of the following groups: (i) control: two placebo lozenges per day for 90 days together with regular oral hygiene, (ii) test: two probiotic lozenges per day for 90 days together with regular oral hygiene. Clinical assessment and biological sample collection will be performed at baseline, 3 and 6 months. In addition, compliance outcomes and adverse events will be monitored.

National audience; In the last decade, the association between the periodontitis and rheumatoid arthritis (RA) has been established, suggesting that oral microbiome plays a causal role by initiating this chronic autoimmune inflammatory disease of articulation. Both pathogenesis are similar in term of chronic inflammation, tissue breakdown and bone resorption. Molecular aspects have also revealed that citrullination, a post-translational modification catalyzed by peptidyl-arginine deiminases (PADs), is involved in both diseases. For RA, citrullinated proteins production leads to the synthesis the of anti-citrullinated protein antibodies triggering the loss of immune tolerance. In humans, five PADs have been identified. Recently, studies have found that only Porphyromonas species possess PAD. Thus, a major periodontal pathogen, Porphyromonas gingivalis, is able to generate citrullinated epitopes, and could consequently induce anti-citrullinated protein antibodies. In this review, citrullination process, periodontitis and RA are described to put them in relation with molecular, clinical and epidemiological studies establishing the association between periodontitis and RA.

The Quebrada de Humahuaca in North West Argentina is a world heritage site listed by the UNESCO since 2003. As pollution is increasing dramatically, a pluridisciplinary, annual-long survey was conducted along the valley to determine dominant patterns in waste management and water quality. Along with the habit of discarding waste into the river, urbanizations have been identified as having a deep influence on the river water quality: iron together with maximum peaks of nitrite and nitrate are related with the anthropogenic activity at Humahuaca, whereas the concentrations of arsenic increase from north to south, with maximum peaks of up to 13.50 and 14.50 μg L−1 measured during the rainy season at Maimará and Volcán, respectively. In this underdeveloped region, economic struggles and waste management customs revealed as major factors causing the environmental degradation. The inclusion in the UNESCO’s sites list was insufficient in preserving this world heritage site. The approach discussed in this article suggests a straightforward analytical measurement of the river hydrochemical properties during the rainy season in arid/semi-arid regions with monsoonal regime, to assess the effects of waste dumping along watercourses. In addition to scientific evaluations, both public acceptance and the establishment of proper waste management infrastructure are fundamental to ensuring the preservation of human and environmental health. Fil: López Steinmetz, Romina Lucrecia. Universidad Nacional de Jujuy. Instituto de Ecorregiones Andinas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Ecorregiones Andinas; Argentina Fil: Fong, Shao Bing. University of Melbourne; Australia Fil: Muhana Senn, Maria Silvina. Gobierno de la Provincia de Jujuy. Ministerio de Infraestructura Servicios Publicos Tierra y Vivienda. Agua Potable de Jujuy Sociedad del Estado.; Argentina Fil: López Steinmetz, Lorena Cecilia. Universidad Nacional de Córdoba. Instituto de Investigaciones Psicológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Psicológicas; Argentina Fil: Meuric, Vincent. Universite de Rennes I; Francia. Inserm; Francia

Vicuna is a wild, endangered species of Andean camelid living in the hyperarid Andean plateau. In the central part of the plateau, the Lithium Triangle defines a zone with lithium‐rich salt pans. Brine pools naturally form within the salt pans, and the adaptation strategy of vicuna consists of drinking from brine pools. Together with reporting the first chemical data on vicuna bones and teeth, we analyzed lithium, boron, and arsenic in water and brines, with the aim of assessing their relation to chronic exposure by water ingestion. We collected and analyzed bones of vicuna specimens lying in an Andean salt pan, together with brine and water samples. Brine and water samples are highly saline and contain large amounts of lithium, boron, and arsenic. Lithium (13.50–40 mg kg–1 ) and boron (40–46.80 mg kg–1 ), but not arsenic, were found in the vicuna bones and teeth. Based on our results and on previously reported data on human tissues in the Andes, we conducted statistical assessments of the relationships between lithium and boron in body tissues and water samples, and discuss their environmental significance in the context of the Lithium Triangle. Fil: López Steinmetz, Romina Lucrecia. Universidad Nacional de Jujuy. Instituto de Ecorregiones Andinas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Ecorregiones Andinas; Argentina. Universidad Nacional de Jujuy. Instituto de Geología Minera; Argentina Fil: Fong, Shao Bing. University of Melbourne; Australia Fil: Boyer, Emile. Universite de Rennes I; Francia Fil: López Steinmetz, Lorena Cecilia. Universidad Nacional de Córdoba. Instituto de Investigaciones Psicológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Psicológicas; Argentina Fil: Tejerina, Norberto Elio. Universidad Nacional de Jujuy. Instituto de Geología Minera; Argentina Fil: Meuric, Vincent. Universite de Rennes I; Francia

International audience; Introduction: Periodontitis is an inflammatory dysbiotic disease. Among putative dysbiosis causes, transmission of Porphyromonas gingivalis between individuals of the same family remains unclear. The aim of this systematic review and meta-analysis is to assess the likelihood of shared detection of Porphyromonas gingivalis among cohabiting family members.Methods: A literature search was conducted on different databases up to September 2018. Articles assessing the presence of P.gingivalis between members of the same family were screened. Only English literature was retrieved, whereas no limits were applied for bacterial sampling and detection methods.Results: Overall, 26 articles published between 1993 and 2017 met the inclusion criteria. Of these, 18 articles were used for meta-analyses. Based on bacterial culture, the likelihood of an intra-familial transmission of P.gingivalis once a member of the family harbors the bacterium is estimated at 63.5% (n = 132 pairs of family members); this drops to 45% when pooling together culture and Polymerase-Chain-Reaction (n = 481 pairs), whereas it is estimated at 35.7% when genotyping is applied (n = 137 pairs).Conclusion: Pooled results suggest that the likelihood of detecting P.gingivalis within within family members is moderately frequent. Personalized periodontal screening and prevention may consider intra-familial co-occurrence of P.gingivalis as feasible.

The use of next generation sequencing and bioinformatics has revealed the complexity and richness of the human oral microbiota. While some species are well known for their periodontal pathogenicity, the molecular-based approaches for bacterial identification have raised awareness about new putative periodontal pathogens. Although they are found increased in case of periodontitis, there is currently a lack of data on their interrelationship with the periodontal measures. We processed the sequencing data of the subgingival microbiota of 75 patients with hemochromatosis and chronic periodontitis in order to characterize the well-described and newly identified subgingival periodontal pathogens. We used correlation tests and statistical models to assess the association between the periodontal pathogens and mean pocket depth, and to determine the most relevant bacterial biomarkers of periodontitis severity. Based on correlation test results, nine taxa were selected and included in the statistical models. The multiple linear regression models adjusted for systemic and periodontal clinical variables showed that mean pocket depth was negatively associated with Aggregatibacter and Rothia, and positively associated with Porphyromonas. Furthermore, a bacterial ratio that was previously described as a signature of dysbiosis in periodontitis (%Porphyromonas+%Treponema+%Tannerella)/(%Rothia+%Corynebacterium) was the most significant predictor. In this specific population, we found that the best model in predicting the mean pocket depth was microbial dysbiosis using the dysbiosis ratio taxa formula. While further studies are needed to assess the validity of these results on the general population, such a dysbiosis ratio could be used in the future to monitor the subgingival microbiota.

International audience; Aim: To investigate the association between periodontal status and serum biomarkers in patients with HFE haemochromatosis.Material and Methods: This clinical case series included 84 HFE-C282Y homozygous patients. Periodontal evaluation was performed using clinical attachment level, probing depth, gingival bleeding index, visible plaque index and gingival index. Serum markers of iron metabolism were collected from medical records. The relationship between serum biomarkers of iron burden and the severity of periodontitis was investigated.Results: The study population consisted of 47 men and 37 women, routinely treated in the Unit of Hepatology, University Hospital, Rennes. All patients presented with periodontitis (mild: n = 1, moderate: n = 37 and severe: n = 46). There was a positive association between transferrin saturation >45% and the severity of periodontitis (adjusted odds ratio = 5.49, p =.002).Conclusion: Severe periodontitis is associated with the severity of iron burden in patients with HFE-related hereditary haemochromatosis. Dental examination should be included in the initial assessment of all these patients. © 2017 John Wiley and Sons A/S. Published by John Wiley and Sons Ltd

International audience; The genus Roseburia consists of obligate Gram-positive anaerobic bacteria that are slightly curved, rod-shaped and motile by means of multiple subterminal flagella. It includes five species: Roseburia intestinalis, R. hominis, R. inulinivorans, R. faecis and R. cecicola. Gut Roseburia spp. metabolize dietary components that stimulate their proliferation and metabolic activities. They are part of commensal bacteria producing short-chain fatty acids, especially butyrate, affecting colonic motility, immunity maintenance and anti-inflammatory properties. Modification in Roseburia spp. representation may affect various metabolic pathways and is associated with several diseases (including irritable bowel syndrome, obesity, Type 2 diabetes, nervous system conditions and allergies). Roseburia spp. could also serve as biomarkers for symptomatic pathologies (e.g., gallstone formation) or as probiotics for restoration of beneficial flora.

After ban of antibiotics growth promoters (AGP) in Europe in 2006, use of non-medicated gut microbiota regulators as feed additives has dramatically increased. This study aimed at describing the effects of a copper-exchanged zeolite on broiler growth performance, small intestine morphology and microbiota composition. Illumina Sequencing of the V3-V4 region of the 16S rRNA gene was employed to study the small intestine microbiota. This microbiota with copper-exchanged zeolite treated-chickens was significantly less diverse with an almost exclusive presence of Lactobacillus johnsonii and Lactobacillus reuteri. These Lactobacilli are correlated with increased diameter, length and weight of the three segments of the small intestine and decreased viscosity of the intestinal content, suggesting probiotic action. The tested copper-exchanged zeolite would act as a prebiotic, selecting a “favorable” flora for the healthy broilers development.

Aim The purpose of this study was to conduct phytochemical analysis of sea buckthorn pulp oil and to evaluate the antimicrobial, anti-biofilm and antioxidant activities of its mouthwash form. Methods and results Fatty acid composition of the sea buckthorn pulp oil was determined by GC-MS analysis, which revealed that, mono-unsaturated fatty acid, palmitoleic acid and saturated fatty acid, palmitic acid, were the major constituents. The antimicrobial and the anti-biofilm capacities of sea buckthorn pulp oil mouthwash form were evaluated against Streptococcus gordonii, Porphyromonas gingivalis, Actinomyces viscosus and Candida albicans, according to the European Norms, and the Biofilm Ring Test® , respectively. These activities were then compared with those of chlorhexidine and herbal mouthwashes. The sea buckthorn-based mouthwash was bactericidal against S. gordonii and P. gingivalis, bacteriostatic against A. viscosus and showed no antifungal effect. Regardless of the strains used, complete inhibition of biofilm formation was achieved. The antioxidant activity of this experimental mouthwash was also assessed by DPPH and NBT assays. Conclusion Sea buckthorn mouthwash showed anti-biofilm activities against select single and multiple oral bacterial species. Significance and impact of the study In this study, a mouthwash derived from sea buckthorn (Hippophae rhamnoides) pulp oil has been experimented, for the first time, in order to overcome the problem of a large number of available synthetic mouthwashes which have side effects on teeth, gums and mucous membranes. This mouthwash seemed to be a suitable alternative for a preventive agent for periodontal inflammation.

International audience; New growth media have been designed for the iron-controlled co-cultures of three oral bacteria. These media share a common core composition enabling the switch from mono- to co-cultures, and efficiently promote both planktonic and biofilm cultures of Porphyromonas gingivalis, Treponema denticola and Streptococcus gordonii.

Periodontitis is driven by disproportionate host inflammatory immune responses induced by an imbalance in the composition of oral bacteria; this instigates microbial dysbiosis, along with failed resolution of the chronic destructive inflammation. The objectives of this study were to identify microbial signatures for health and chronic periodontitis at the genus level and to propose a model of dysbiosis, including the calculation of bacterial ratios. Published sequencing data obtained from several different studies (196 subgingival samples from patients with chronic periodontitis and 422 subgingival samples from healthy subjects) were pooled and subjected to a new microbiota analysis using the same Visualization and Analysis of Microbial Population Structures (VAMPS) pipeline, to identify microbiota specific to health and disease. Microbiota were visualized using CoNet and Cytoscape. Dysbiosis ratios, defined as the percentage of genera associated with disease relative to the percentage of genera associated with health, were calculated to distinguish disease from health. Correlations between the proposed dysbiosis ratio and the periodontal pocket depth were tested with a different set of data obtained from a recent study, to confirm the relevance of the ratio as a potential indicator of dysbiosis. Beta diversity showed significant clustering of periodontitis-associated microbiota, at the genus level, according to the clinical status and independent of the methods used. Specific genera ( Veillonella , Neisseria , Rothia , Corynebacterium , and Actinomyces ) were highly prevalent (>95%) in health, while other genera ( Eubacterium , Campylobacter , Treponema , and Tannerella ) were associated with chronic periodontitis. The calculation of dysbiosis ratios based on the relative abundance of the genera found in health versus periodontitis was tested. Nonperiodontitis samples were significantly identifiable by low ratios, compared to chronic periodontitis samples. When applied to a subgingival sample set with well-defined clinical data, the method showed a strong correlation between the dysbiosis ratio, as well as a simplified ratio ( Porphyromonas , Treponema , and Tannerella to Rothia and Corynebacterium ), and pocket depth. Microbial analysis of chronic periodontitis can be correlated with the pocket depth through specific signatures for microbial dysbiosis. IMPORTANCE Defining microbiota typical of oral health or chronic periodontitis is difficult. The evaluation of periodontal disease is currently based on probing of the periodontal pocket. However, the status of pockets “on the mend” or sulci at risk of periodontitis cannot be addressed solely through pocket depth measurements or current microbiological tests available for practitioners. Thus, a more specific microbiological measure of dysbiosis could help in future diagnoses of periodontitis. In this work, data from different studies were pooled, to improve the accuracy of the results. However, analysis of multiple species from different studies intensified the bacterial network and complicated the search for reproducible microbial signatures. Despite the use of different methods in each study, investigation of the microbiota at the genus level showed that some genera were prevalent (up to 95% of the samples) in health or disease, allowing the calculation of bacterial ratios (i.e., dysbiosis ratios). The correlation between the proposed ratios and the periodontal pocket depth was tested, which confirmed the link between dysbiosis ratios and the severity of the disease. The results of this work are promising, but longitudinal studies will be required to improve the ratios and to define the microbial signatures of the disease, which will allow monitoring of periodontal pocket recovery and, conceivably, determination of the potential risk of periodontitis among healthy patients.

International audience; Periodontitis are bacterial inflammatory diseases, where the bacterial biofilms present on the tooth-supporting tissues switch from a healthy state towards a pathogenic state. Among bacterial species involved in the disease, Porphyromonas gingivalis has been shown to induce dysbiosis, and to induce virulence of otherwise healthy bacteria like Streptococcus gordonii. During biofilm development, primary colonizers such as S. gordonii first attach to the surface and allow the subsequent adhesion of periodontal pathogens such as P. gingivalis. Interactions between those two bacteria have been extensively studied during the adhesion step of the biofilm. The aim of the study was to understand interactions of both species during the growing phase of the biofilm, for which little knowledge is available, using a mathematical model. This two-species biofilm model was based on a substrate-dependent growth, implemented with damage parameters, and validated thanks to data obtained on experimental biofilms. Three different hypothesis of interactions were proposed and assayed using this model: independence, competition between both bacteria species, or induction of toxicity by one species for the other species. Adequacy between experimental and simulated biofilms were found with the last hypothetic mathematical model. This new mathematical model of two species bacteria biofilms, dependent on different substrates for growing, can be applied to any bacteria species, environmental conditions, or steps of biofilm development. It will be of great interest for exploring bacterial interactions in biofilm conditions.

International audience; For millions of years, our resident microbes have coevolved and coexisted with us in a mostly harmonious symbiotic relationship. We are not distinct entities from our microbiome, but together we form a 'superorganism' or holobiont, with the microbiome playing a significant role in our physiology and health. The mouth houses the second most diverse microbial community in the body, harbouring over 700 species of bacteria that colonise the hard surfaces of teeth and the soft tissues of the oral mucosa. Through recent advances in technology, we have started to unravel the complexities of the oral microbiome and gained new insights into its role during both health and disease. Perturbations of the oral microbiome through modern-day lifestyles can have detrimental consequences for our general and oral health. In dysbiosis, the finely-tuned equilibrium of the oral ecosystem is disrupted, allowing disease-promoting bacteria to manifest and cause conditions such as caries, gingivitis and periodontitis. For practitioners and patients alike, promoting a balanced microbiome is therefore important to effectively maintain or restore oral health. This article aims to give an update on our current knowledge of the oral microbiome in health and disease and to discuss implications for modern-day oral healthcare.

Transient bacteremia from oral cavity related to oral anaerobic bacteria may occur as a result of dental healthcare procedures but also as a result of daily gestures involving the gums (chewing and oral hygiene). The risk of presenting a transient bacteremia is related to oral cavity bacterial load and to the severity of inflammation in the oral cavity. Although bacteremia is transient, in patients with immunodeficiency or comorbidity, this bacteremia may cause extra-oral infections. The bacteremia rate and the identified bacteria vary from one study to the next, depending on the method used to isolate and identify bacteria. Nevertheless, the risk for bacteremia is determined by the infectious and inflammatory conditions of each patient.

Introduction: Porphyromonas gingivalis, a gram-negative anaerobic bacterium, is a major periopathogen whose transmission from host to host involves exposure to atmospheric oxygen. P. gingivalis contains genetic factors that function in an oxidative stress response, but their expression has not been analyzed during exposure to atmospheric oxygen. The aim of this study was to obtain a better understanding of atmospheric adaptation of P. gingivalis. Methods: The aerotolerance of wild-type and oxyR mutant P. gingivalis strains were determined, and quantitative polymerase chain reaction was performed to analyze gene expression patterns in response to exposure to atmospheric oxygen. The analyzed P. gingivalis genes encoded proteins involved in oxidative response (oxyR, ahpC-F, batA, dps, ftn, tpx) as well as several major virulence factors (hagA, hagB, hagE, rgpA, rgpB, hem). Results: Our results demonstrated a critical role for the oxyR gene in the aerotolerance of P. gingivalis. The ahpC-F, batA, and hem genes were slightly overexpressed (between 1.65-fold and 2-fold) after exposure to atmospheric oxygen compared to anaerobic conditions. The level of transcription of dps, ftn, tpx, and rgpA genes increased more than 2.5-fold, and the expression of ahpC-F, dps, ftn, and tpx was partially or completely OxyR-dependent. Conclusion: A different transcription pattern of P. gingivalis genes was observed, depending on the stimulus of oxidative stress. We present new evidence that the expression of tpx, encoding a thiol peroxidase, is partially OxyR-dependent and is induced after atmospheric oxygen exposure.

Meuric EA1254; International audience; Porphyromonas gingivalis is a major pathogen associated with chronic periodontitis. The organism’s cell-surface cysteine proteinases, the Arg-specific proteinases (RgpA, RgpB) and the Lys-specific proteinase (Kgp), which are known as gingipains have been implicated as major virulence factors. All three gingipain precursors contain a propeptide of around 200 amino acids in length that is removed during maturation. The aim of this study was to characterize the inhibitory potential of the Kgp and RgpB propeptides against the mature cognate enzymes. Mature Kgp was obtained from P. gingivalis mutant ECR368, which produces a recombinant Kgp with an ABM1 motif deleted from the catalytic domain (rKgp) that enables the otherwise membrane bound enzyme to dissociate from adhesins and be released. Mature RgpB was obtained from P. gingivalis HG66. Recombinant propeptides of Kgp and RgpB were produced in Escherichia coli and purified using nickel-affinity chromatography. The Kgp and RgpB propeptides displayed non-competitive inhibition kinetics with Ki values of 2.04 µM and 12 nM, respectively. Both propeptides exhibited selectivity towards their cognate proteinase. The specificity of both propeptides was demonstrated by their inability to inhibit caspase-3, a closely related cysteine protease, and papain that also has a relatively long propeptide. Both propeptides at 100 mg/L caused a 50% reduction of P. gingivalis growth in a protein-based medium. In summary, this study demonstrates that gingipain propeptides are capable of inhibiting their mature cognate proteinases

International audience; Porphyromonas gingivalis, an important etiological agent of periodontal disease, is frequently found associated with Treponema denticola, an anaerobic spirochete, in pathogenic biofilms. However, interactions between these two bacteria are not well understood at the molecular level. In this study, we seek to link the influence of T. denticola on the expression of P. gingivalis proteases with its capacities to adhere and to form biofilms. The P. gingivalis genes encoding Arg-gingipain A (RgpA), Lys-gingipain (Kgp), and hemagglutinin A (HagA) were more strongly expressed after incubation with T. denticola compared with P. gingivalis alone. The amounts of the three resulting proteins, all of which contain hemagglutinin adhesion domains, were increased in culture supernatants. Moreover, incubation of P. gingivalis with T. denticola promoted static and dynamic biofilm formation, primarily via a time-dependent enhancement of P. gingivalis adhesion capacities on bacterial partners such as Streptococcus gordonii. Adhesion of P. gingivalis to human cells was also increased. These results showed that interactions of P. gingivalis with other bacterial species, such as T. denticola, induce increased adhesive capacities on various substrata by hemagglutinin adhesion domain-containing proteins.

International audience; Porphyromonas gingivalis, an anaerobic, asaccharolytic gram-negative bacterium, is a causative agent in chronic periodontitis. It has many virulence factors that facilitate infection of the gingiva, but little is known about the local immune cells that respond to this bacterium. The aims of this study were to quantify P. gingivalis in gingival biopsies from patients with periodontitis using laser capture microdissection (LCM) plus qRT-PCR and to determine the phenotype of immune cells associated with the bacteria using immunofluorescence. The presence of P. gingivalis was confirmed in periodontitis gingival tissue from 10 patients, and differences in bacterial distribution in the epithelium and connective tissue with or without inflammatory infiltrates were observed. Immune cells found in the biopsy tissues, including CD20+ mature B cells and CD138+ plasma cells, were associated with the Th2-type immune response. Most P. gingivalis was in direct contact with CD4+ T cells. This study revealed for the first time the colocalization of P. gingivalis with immune cells. Use of LCM combined with qRT-PCR enabled quantitative analysis of bacteria in a selected area of a biopsy sample without any tissue degradation. Observation of the immune cells associated with these bacteria was also performed by immunofluorescence.

Oral Communication presented at the ";Forum des Jeunes Chercheurs";, Brest (France) 2011.

International audience; The electron transfer chain in Porphyromonas gingivalis, or periodontopathogens, has not yet been characterized. P. gingivalis, a strict anaerobic bacteria and the second colonizer of the oral cavity, is considered to be a major causal agent involved in periodontal diseases. Primary colonizers create a favorable environment for P. gingivalis growth by decreasing oxygen pressure. Oxygen does not appear to be the final electron acceptor of the respiratory chain. Fumarate and cytochrome b have been implicated as major components of the respiratory activity. However, the P. gingivalis genome shows many other enzymes that could be implicated in aerobic or nitrite respiration. Using bioinformatic tools and literature studies of respiratory pathways, the ATP synthesis mechanism from the sodium cycle and nutrients metabolism, the putative respirasome of P. gingivalis has been proposed.

Porphyromonas gingivalis is considered as a major etiological agent in the onset and progression of chronic destructive periodontitis. Porphyromonus gingivalis fimA type has been correlated to the virulence potential of the strain; therefore this gene could be involved in the ability of P. gingivalis to reach blood stream.The classifications of P. gingivalis fimA types will be compared in subgingival plaque and blood samples collected after scaling and root root planing of periodontitis patients.Fifteen periodontitis patients requiring scaling and root planing were enrolled. P. gingivalis isolates were classed to genotype with fimA type-specific PCR assay. fimA gene was sequenced if the isolate was listed as unclassifiable after PCR technique.Six patients showed positive P. gingivalis bacteremia. The most frequent fimA was fimA type II, followed by Ib, III and IV. In blood strains, type II was followed by IV, Ib and III.Type II was the most frequent genotype in blood samples and in subgingival plaque samples. However, no correlation was found between the frequency of any fimA type with SRP induced bacteremia. P. gingivalis fimA type appears to be conserved within individual patients throughout the times of sample collection, fimA gene sequence results were not in agreement with results of fimA genotyping by PCR.

International audience; Porphyromonas gingivalis is an anaerobic periodontal pathogen that resides in the complex multispecies microbial biofilm known as dental plaque. Effective reporter tools are increasingly needed to facilitate physiological and pathogenetic studies of dental biofilm. Fluorescent proteins are ideal reporters for conveniently monitoring biofilm growth, but are restricted by several environmental factors, such as a requirement of oxygen to emit fluorescence. We developed a fluorescent reporter plasmid, known as the SNAP-tag, for labeling P. gingivalis cells, which encode an engineered version of the human DNA repair enzyme O(6)-alkylguanine-DNA alkyltransferase. Fluorescent substrates containing O(6)-benzylguanine covalently and specifically bind to the enzyme via stable thioether bonds. For the present study, we constructed a replicative plasmid carrying SNAP26b under the control of the P. gingivalis endogenous trxB promoter. The P. gingivalis-expressing SNAP26 protein was successfully labeled with specific fluorophores under anaerobic conditions. Porphyromonas gingivalis biofilm formation was investigated using flow cells and confocal laser scanning microscopy. A specific distribution of a strong fluorescence signal was demonstrated in P. gingivalis-SNAP26 monospecies and bispecies biofilms with Streptococcus gordonii-GFPmut3(*). These findings show that the SNAP-tag can be applied to studies of anaerobic bacteria in biofilm models and is a useful and advantageous alternative to existing labeling strategies.

Porphyromonas gingivalis and Treponema denticola are strongly associated with chronic periodontitis. These bacteria have been co-localized in subgingival plaque and demonstrated to exhibit symbiosis in growth in vitro and synergistic virulence upon co-infection in animal models of disease. Here we show that during continuous co-culture a P. gingivalis:T. denticola cell ratio of 6∶1 was maintained with a respective increase of 54% and 30% in cell numbers when compared with mono-culture. Co-culture caused significant changes in global gene expression in both species with altered expression of 184 T. denticola and 134 P. gingivalis genes. P. gingivalis genes encoding a predicted thiamine biosynthesis pathway were up-regulated whilst genes involved in fatty acid biosynthesis were down-regulated. T. denticola genes encoding virulence factors including dentilisin and glycine catabolic pathways were significantly up-regulated during co-culture. Metabolic labeling using 13C-glycine showed that T. denticola rapidly metabolized this amino acid resulting in the production of acetate and lactate. P. gingivalis may be an important source of free glycine for T. denticola as mono-cultures of P. gingivalis and T. denticola were found to produce and consume free glycine, respectively; free glycine production by P. gingivalis was stimulated by T. denticola conditioned medium and glycine supplementation of T. denticola medium increased final cell density 1.7-fold. Collectively these data show P. gingivalis and T. denticola respond metabolically to the presence of each other with T. denticola displaying responses that help explain enhanced virulence of co-infections., Author Summary Unlike the traditional view that most diseases are caused by infection with a single bacterial species, some chronic diseases including periodontitis result from the perturbation of the natural microbiota and the proliferation of a number of opportunistic pathogens. Both Porphyromonas gingivalis and Treponema denticola have been associated with the progression and severity of chronic periodontitis and have been shown to display synergistic virulence in animal models. However, the underlying mechanisms to these observations are unclear. Here we demonstrate that these two bacteria grow synergistically in continuous co-culture and modify their gene expression. The expression of T. denticola genes encoding known virulence factors and enzymes involved in the uptake and metabolism of the amino acid glycine was up-regulated in co-culture. T. denticola stimulated the proteolytic P. gingivalis to produce free glycine, which T. denticola used as a major carbon source. Our study shows P. gingivalis and T. denticola co-operate metabolically and this helps to explain their synergistic virulence in animal models and their intimate association in vivo.

Introduction. Porphyromonas gingivalis is considered as a major etiological agent in the onset and progression of chronic destructive periodontitis. Porphyromonus gingivalis fimA type has been correlated to the virulence potential of the strain; therefore this gene could be involved in the ability of P. gingivalis to reach blood stream. Objective. The classifications of P. gingivalis fimA types will be compared in subgingival plaque and blood samples collected after scaling and root root planing of periodontitis patients. Materials and methods. Fifteen periodontitis patients requiring scaling and root planing were enrolled. P. gingivalis isolates were classed to genotype with fimA type-specific PCR assay. fimA gene was sequenced if the isolate was listed as unclassifiable after PCR technique. Results. Six patients showed positive P. gingivalis bacteremia. The most frequent fimA was fimA type II, followed by Ib, III and IV. In blood strains, type II was followed by IV, Ib and III. Conclusion. Type II was the most frequent genotype in blood samples and in subgingival plaque samples. However, no correlation was found between the frequency of any fimA type with SRP induced bacteremia. P. gingivalis fimA type appears to be conserved within individual patients throughout the times of sample collection. fimA gene sequence results were not in agreement with results of fimA genotyping by PCR. Introducción. Porphyromonas gingivalis es el principal agente etiológico de la periodontitis. El gen fimA ha sido relacionado con la virulencia del microorganismo, lo cual sugiere la participación de dicho gen en la capacidad del microorganismo para alcanzar el torrente sanguíneo. Objetivo. Estudiar la distribución de los tipos de fimA de P. gingivalis en muestras de placa subgingival y de sangre obtenidas durante bacteriemias después de raspaje y alisado radicular. Materiales y métodos. Se practicó un alisado radicular a 15 pacientes con periodontitis. Se obtuvieron aislamientos clínicos de P. gingivalis de la placa subgingival y durante la bacteriemia inducida por el procedimiento. Para la genotipificación se utilizó la técnica de reacción en cadena de la polimerasa (PCR) específica para fimA. En los aislamientos no clasificables por PCR se realizó secuenciación del gen fimA. Resultados. Seis pacientes fueron positivos para bacteriemia por P. gingivalis. La distribución de fimA evaluada en 30 aislamientos de placa subgingival y de sangre mostró una mayor frecuencia del fimA tipo II de P. gingivalis. En los aislamientos de placa subgingival, la detección de fimA tipo II fue seguida por Ib, III y IV; sin embargo, en los aislamientos de sangre el tipo II fue seguido por los tipos IV, Ib y III. Conclusión. En los aislamientos de sangre y de placa subgingival de pacientes con periodontitis el fimA más frecuente fue el tipo II; no fue posible correlacionar el tipo de fimA con la bacteriemia inducida por el alisado radicular. Los resultados de la secuenciación del gen fimA no concuerdan con los obtenidos por PCR.

International audience

In the oral cavity, periodontal health relies on a balance between the subgingival bacterial community and the host’s immune function. The current pathophysiological model of chronic periodontitis shows a negative feedback loop that implies a dysbiotic microbiota and a dysregulation of the inflammatory response. As an increasing number of associations between the periodontal disease and systemic conditions are reported, this work describes how a genetic iron overload disease — HFE-related hereditary haemochromatosis — is likely to influence the host–microbiota interactions within the subgingival sulcus. We conducted clinical and microbiological cross-sectional studies, that showed transferrin saturation coefficient having an impact on the severity of periodontitis, probably related to the promotion of periodontal pathogenic species. However, morphometric analysis of the murine model for hereditary haemochromatosis Hfe(–/–) revealed alterations in the mandibular alveolar bone micro-architecture, suggesting a disruption of bone remodeling. A mixed animal model, with Porphyromonas gingivalis-induced periodontitis and Hfe(–/–) mice, was therefore performed, in order to explore the underlying mechanisms of the relationships between iron excess, bacterial dysbiosis, and periodontitis.; Dans la cavité buccale, la santé parodontale repose sur un équilibre entre la communauté bactérienne sous-gingivale et la fonction immunitaire de l’hôte, toutes deux compatibles avec un état sain. Le modèle pathophysiologique actuel de la parodontite chronique montre un cercle vicieux entre un microbiote dysbiotique et une dérégulation de la réponse inflammatoire. Alors qu’un nombre croissant d’associations entre maladie parodontale et pathologie systémique sont rapportées, ce travail décrit comment une maladie génétique de surcharge en fer — l’hémochromatose héréditaire liée au gène HFE — est susceptible d’influencer l’équilibre hôte–microbiote au sein du sillon sous-gingival. Les études transversales, clinique et microbiologique, que nous avons menées, ont ainsi montré un impact du coefficient de saturation de la transferrine sur la sévérité de la parodontite, probablement en lien avec la sélection d’espèces pathogènes pour le parodonte. Cependant, l’analyse morphométrique du modèle murin de l’hémochromatose héréditaire Hfe(–/–) a révélé des altérations de la micro-architecture de l’os alvéolaire mandibulaire, suggérant un dérèglement du remodelage osseux. Un croisement entre deux modèles animaux, parodontite induite par Porphyromonas gingivalis et Hfe(–/–), a donc été mis en œuvre, afin d’explorer les mécanismes de cette association entre excès de fer, dysbiose bactérienne et parodontite.