Metabolomic Signatures of In Vitro Biofilm Maturation of Streptococcus mutans.

The Streptococcus mutans is commonly find in oral environment in both symbiont and dysbiotic conditions, where for the last one it causes the break in homeostatic balance and, in association with other microorganisms' community, results in dental caries process. Additionally, it is important to determine the low molecular weight metabolites profile from Streptococcus mutans to distinguish the endogenous and exogenous compounds from patient subjected to salivary metabolomic studies. Thus, the objective of the present study was to characterize the in vitro metabolomic profile of the maturation of a single-species Streptococcus mutans biofilm using metabolomic approach by 1H-nuclear magnetic resonance (NMR) spectroscopy. A distinct metabolomic profile was observed after 2 days of biofilm maturation, independently of the presence of enamel substrate. Sucrose, lactate, and fructose were the main metabolites responsible for the distinction. The sucrose was consumed by S. mutans in higher levels in the initial experimental periods than at 6 days of biofilm growth. Lactate and fructose were the main compounds secreted, regardless of the type of growth, but it was also observed production of propionate, iso-butyrate, and pyruvate. Pyruvate metabolism and glycolysis/gluconeogenesis were the main pathways related to biofilm growth. The results contribute to the determination of compounds that are resulted from oral microbial activity and help to guide further metabolomics studies.

Impact of Precompetitive Training on Metabolites in Modern Pentathletes.

PURPOSE: Modern pentathlon is a multidisciplinary sport that involves exhaustive training which can cause tissue damage and metabolic changes. However, few studies have evaluated the metabolic changes that occur in pentathletes. Accordingly, we aimed to evaluate the metabolomic profile of pentathletes during a 3-week training period before competition using nuclear magnetic resonance. METHODS: Blood samples from 6 members of a Brazilian modern pentathlon team were collected at the beginning (Pre1, Pre2, and Pre3) and end (Post1, Post2, and Post3) of each week. Low molecular-weight metabolite profiles were analyzed by nuclear magnetic resonance spectroscopy, and biochemical markers were assessed using enzyme-linked immunosorbent assays. Data were assessed using partial least-squares discriminant analysis and univariate statistical model. RESULTS: Metabolic changes were observed between pre- and postdata of each week and over the 3 weeks before the competition in the partial least-squares discriminant analysis. Creatine kinase concentration increased in the first 2 weeks (P = .045 and P = .039), but there was no difference in the last week (P > .05). Lactate levels increased significantly after training in each week (P < .001). Cortisol levels at Post3 were different from all other time points (P < .05) and the concentrations of peroxides increased over the weeks (P < .05). Among all metabolites, sarcosine showed the greatest differences (P = .004) in the pretraining and posttraining periods of the 3 weeks. CONCLUSION: Serum analysis of athletes using nuclear magnetic resonance showed metabolic changes depending on the intensity of the training performed each week.

The bacterial microbiome and metabolome in caries progression and arrest.

Aim: This in vivo experimental study investigated bacterial microbiome and metabolome longitudinal changes associated with enamel caries lesion progression and arrest. Methods: We induced natural caries activity in three caries-free volunteers prior to four premolar extractions for orthodontic reasons. The experimental model included placement of a modified orthodontic band on smooth surfaces and a mesh on occlusal surfaces. We applied the caries-inducing protocol for 4- and 6-weeks, and subsequently promoted caries lesion arrest via a 2-week toothbrushing period. Lesions were verified clinically and quantitated via micro-CT enamel density measurements. The biofilm microbial composition was determined via 16S rRNA gene Illumina sequencing and NMR spectrometry was used for metabolomics. Results: Biofilm maturation and caries lesion progression were characterized by an increase in Gram-negative anaerobes, including Veillonella and Prevotella. Streptococcus was associated caries lesion progression, while a more equal distribution of Streptococcus, Bifidobacterium, Atopobium, Prevotella, Veillonella, and Saccharibacteria (TM7) characterized arrest. Lactate, acetate, pyruvate, alanine, valine, and sugars were more abundant in mature biofilms compared to newly formed biofilms. Conclusions: These longitudinal bacterial microbiome and metabolome results provide novel mechanistic insights into the role of the biofilm in caries progression and arrest and offer promising candidate biomarkers for validation in future studies.

Salivary metabolome of children and adolescents under peritoneal dialysis.

OBJECTIVE: To study the influence of peritoneal dialysis (PD) on the salivary metabolite profile of children and adolescents with renal failure. MATERIALS AND METHODS: Healthy children/adolescents (n = 31; mean age: 12.18 ± 3.76) and children/adolescents subjected to PD (n = 12; mean age: 10.10 ± 4.25) were recruited. Oral health status assessed by the dmft/DMFT and Volpe-Manhold calculus indices. The 1H spectra were acquired in a 600-MHz Bruker nuclear magnetic resonance spectrometer and were subjected to multivariate analysis using partial least squares discriminant analysis (PLS-DA), orthogonal PLS-DA (O-PLS-DA), and univariate analysis through chi-square and t tests (SPSS 20.0, IL, USA), with a significance level of p < 0.05. RESULTS: A similar caries pattern (p = 0.57; chi-square test) was observed between the healthy (dmft = 0.72 ± 1.28 and DMFT 0.93 ± 2.30) and PD groups (dmft = 2.14 ± 3.67, DMFT 0.33 ± 0.71) and dental calculus (p > 0.05, t test). PLS-DA and O-PLS-DA were able to distinguish both groups (ACC = 0.85, R2 = 0.80, Q2 = 0.15). Salivary metabolites decrease in creatine, propionate, and sugar levels in the PD group and an increase in creatinine, butyrate, and lactate levels when compared with the healthy group. CONCLUSIONS: Children and adolescents subjected to PD have a different salivary metabolic profile from that of their healthy subjects. CLINICAL RELEVANCE: Complications of peritoneal dialysis procedures could be monitored by proper knowledge of saliva characteristics as predictors of peritonitis-related outcome. The use of metabolomics in pediatric nephrology may be an innovative methodology for the early diagnosis and monitoring of kidney diseases.

The efficacy of transcranial direct current stimulation and transcranial magnetic stimulation for chronic orofacial pain: A systematic review.

BACKGROUND: Transcranial Direct Current Stimulation (tDCS) and Transcranial Magnetic Stimulation (TMS) have been described as promising alternatives to treat different pain syndromes. This study evaluated the effects of TMS and tDCS in the treatment of chronic orofacial pain, through a systematic review. METHODS: An electronic search was performed in major databases: MEDLINE, Scopus, Web of Science, Cochrane, Embase, LILACS, BBO, Open Gray and CINAHL. The eligibility criteria comprised randomized clinical trials (RCTs) that applied TMS or tDCS to treat chronic orofacial pain. The variables analyzed were pain, functional limitation, quality of life, tolerance to treatment, somatosensory changes, and adverse effects. The risk of bias was assessed through the Cochrane Collaboration tool, and the certainty of evidence was evaluated through GRADE. The protocol was registered in the PROSPERO database (CRD42018090774). RESULTS: The electronic search resulted in 636 studies. Thereafter, the eligibility criteria were applied and the duplicates removed, resulting in eight RCTs (four TMS and four tDCS). The findings of these studies suggest that rTMS applied to the Motor cortex (M1), the dorsolateral prefrontal cortex (DLPFC) and the secondary somatosensory cortex (S2) provide adequate orofacial pain relief. Two studies reported significant pain improvement with tDCS applied over M1 while the other two failed to demonstrate significant effects compared to placebo. CONCLUSIONS: rTMS, applied to M1, DLPFC or S2, is a promising approach for the treatment of chronic orofacial pain. Moreover, tDCS targeting M1 seems to be also effective in chronic orofacial pain treatment. The included studies used a wide variety of therapeutic protocols. In addition, most of them used small sample sizes, with a high risk of biases in their methodologies, thus producing a low quality of evidence. The results indicate that further research should be carried out with caution and with better-standardized therapeutic protocols.

Effectiveness of varnish with CPP-ACP in prevention of caries lesions around orthodontic brackets: an OCT evaluation.

OBJECTIVE: To evaluate the in vitro efficiency of applying varnish containing casein phosphopeptide (CPP) and amorphous calcium phosphate (ACP) in prevention of caries lesions around orthodontic brackets. MATERIALS AND METHODS: For this purpose, brackets were bonded to the vestibular surface of bovine incisors, and eight groups were formed (n = 15) according to exposure of oral hygiene substances and enamel varnish: 1 (control) brushing only performed, 2 (control) brushing + use of mouth wash with fluoride, 3 Duraphat varnish application only (Colgate-Palmolive Ind. E Com. Ltda, São Paulo, SP, Brazil), 4 Duraphat + brushing, 5 Duraphat + brushing + mouth wash, 6 MI Varnish application (GC America, USA), 7 MI + brushing, and 8 MI + brushing + mouth wash. The experimental groups alternated between pH cycling and the procedures described and were kept in an oven at temperature of 37°C. Both brushing and immersion in solutions was performed in a time interval of 1 minute, followed by washing in deionized water three times a day for 28 days of experimentation. Afterwards, evaluation by optical coherence tomography (OCT) of the special type (Ganymede OCT/Thorlabs, Newton, USA) was performed. In each group, a scanning exam of the white spot lesion area (around the region where brackets were bonded) and depth measurement of caries lesions were performed. RESULTS: Groups 1 and 3 were shown to differ statistically from groups 5, 6, 7, and 8 (p = 0.000). MI Varnish was shown to be more effective in diminishing caries lesion depth, compared with Duraphat, irrespective of being associated with brushing and mouth wash, or not. LIMITATIONS: The major limitation of this study is that it is a study in which demineralization was obtained with the use of chemical products, and did not occur due to the presence of Streptococcus mutans and its acid byproducts. CONCLUSION: Application CPP-ACP-containing varnish irrespective of being associated with brushing and mouthwash, or not, reduced depth of caries lesions around orthodontic brackets.