Anti-Inflammatory Effect of Caffeine on Muscle under Lipopolysaccharide-Induced Inflammation.

Evidence has shown that caffeine administration reduces pro-inflammatory biomarkers, delaying fatigue and improving endurance performance. This study examined the effects of caffeine administration on the expression of inflammatory-, adenosine receptor- (the targets of caffeine), epigenetic-, and oxidative metabolism-linked genes in the vastus lateralis muscle of mice submitted to lipopolysaccharide (LPS)-induced inflammation. We showed that caffeine pre-treatment before LPS administration reduced the expression of Il1b, Il6, and Tnfa, and increased Il10 and Il13. The negative modulation of the inflammatory response induced by caffeine involved the reduction of inflammasome components, Asc and Casp1, promoting an anti-inflammatory scenario. Caffeine treatment per se promoted the upregulation of adenosinergic receptors, Adora1 and Adora2A, an effect that was counterbalanced by LPS. Moreover, there was observed a marked Adora2A promoter hypermethylation, which could represent a compensatory response towards the increased Adora2A expression. Though caffeine administration did not alter DNA methylation patterns, the expression of DNA demethylating enzymes, Tet1 and Tet2, was increased in mice receiving Caffeine+LPS, when compared with the basal condition. Finally, caffeine administration attenuated the LPS-induced catabolic state, by rescuing basal levels of Ampk expression. Altogether, the anti-inflammatory effects of caffeine in the muscle can be mediated by modifications on the epigenetic landscape.

Impact of neuroinflammation on epigenetic transcriptional control of Sonic Hedgehog members in the central nervous system.

Sonic Hedgehog (Shh) signaling plays a critical role during central nervous system (CNS) development, and its dysregulation leads to neurological disorders. Nevertheless, little is known about Shh signaling regulation in the adult brain. Here, we investigated the contribution of DNA methylation on the transcriptional control of Shh signaling pathway members and its basal distribution impact on the brain, as well as its modulation by inflammation. The methylation status of the promoter regions of these members and the transcriptional profile of DNA-modifying enzymes (DNA Methyltransferases - DNMTs and Tet Methylcytosine Dioxygenase - TETs) were investigated in a murine model of neuroinflammation by qPCR. We showed that, in the adult brain, methylation in the CpG promoter regions of the Shh signaling pathway members was critical to determine the endogenous differential transcriptional pattern observed between distinct brain regions. We also found that neuroinflammation differentially modulates gene expression of DNA-modifying enzymes. This study reveals the basal transcriptional profile of DNMTs and TETs enzymes in the CNS and demonstrates the effect of neuroinflammation on the transcriptional control of members of the Shh Signaling pathway in the adult brain.

Sampling strategies for wastewater surveillance: Evaluating the variability of SARS-COV-2 RNA concentration in composite and grab samples.

The shedding of SARS-CoV-2 RNA titers by infected individuals, even asymptomatic and oligosymptomatic ones, allows the use of wastewater monitoring to track the COVID-19 spread in a community. This approach is interesting especially for emerging countries with limited clinical testing capabilities. However, there are still important methodological aspects that need validation so that wastewater monitoring data become more representative and useful for public health. This study evaluated the between-day and within-day variability of SARS-CoV-2 RNA concentrations in 24-hour composite and grab samples from three different sampling points, including two wastewater treatment plants (WTTP) and a sewer manhole. In the between-day evaluation (17 weeks of monitoring), a good agreement between the SARS-CoV-2 RNA concentration of each sampling method was observed. There were no significant differences between the mean concentrations of the grab and composite samples (p-value > 0.05), considering N1 and N2 gene assays. The strong relationship between composite and grab samples was proven by correlation coefficients: Pearson's r of 0.83 and Spearman's rho of 0.78 (p-value < 0.05). In within-day evaluation, 24-hour cycles were analyzed and low variability in hourly viral concentrations was observed for three sampling points. The coefficient of variation (CV) values ranged from 3.0% to 11.5%. Overall, 24-hour profiles showed that viral RNA concentrations had less variability and greater agreement with the mean values between 8 a.m. and 10 a.m, the recommended time for grab sampling. Therefore, this study provides important information on wastewater sampling techniques for COVID-19 surveillance. Wastewater monitoring information will only be useful to public health and decision-makers if we ensure data quality through best practices.

Non-coding RNAs repressive role in post-transcriptional processing of RUNX2 during the acquisition of the osteogenic phenotype of periodontal ligament mesenchymal stem cells.

Mesenchymal stem cells are candidates for therapeutic strategies in periodontal repair due to their osteogenic potential. In this study, we identified epigenetic markers during osteogenic differentiation, taking advantage of the individual pattern of mesenchymal cells of the periodontal ligament with high (h-PDLCs) and low (l-PDLCs) osteogenic capacity. We found that the involvement of non-coding RNAs in the regulation of the RUNX2 gene is strongly associated with high osteogenic potential. Moreover, we evaluated miRs and genes that encode enzymes to process miRs and their biogenesis. Our data show the high expression of the XPO5 gene, and miRs 7 and 22 observed in the l-PDLCs might be involved in acquiring osteogenic potential, suppressing RUNX2 gene expression. Further, an inversely proportional correlation between lncRNAs (HOTAIR and HOTTIP) and RUNX2 gene expression was observed in both l- and h-PDLCs, and it was also related to the distinct osteogenic phenotypes. Thus, our results indicate the low expression of XPO5 in h-PDLC might be the limiting point for blocking the miRs biogenesis, allowing the high gene expression of RUNX2. In accordance, the low expression of miRs, HOTAIR, and HOTTIP could be a prerequisite for increased osteogenic potential in h-PDLCs. These results will help us to better understand the underlying mechanisms of osteogenesis, considering the heterogeneity in the osteogenic potential of PDLCs that might be related to a distinct transcriptional profile of lncRNAs and the biogenesis machinery.

Preoperative Mouthwash in Subjects with Different Periodontal Status: A Randomised Controlled Clinical Trial.

PURPOSE: The effects of three preoperative mouthwashes on salivary bacterial levels were evaluated and compared between subjects with differing periodontal status. MATERIALS AND METHODS: Based on periodontal parameters, periodontally healthy individuals (n = 60) and those with gingivitis (n = 60) and periodontitis (n = 60) were randomly assigned to a single preoperative dose of chlorhexidine (CHX), essential oils (EO), cetylpyridinium chloride (CPC) or negative control mouthwashes. Saliva samples were collected between 8:00 and 11:00 a.m., before and after a single-dose rinse with the respective mouthwash. Total bacterial load and levels of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and Streptococcus oralis were determined by qPCR. Data were statistically analysed using paired t- and Student's t-tests (p < 0.05). RESULTS: CHX, EO and CPC showed greater antimicrobial efficacy than did the negative control. CHX [1226445.53] and EO [1743639.38] provided greater reductions in comparison to both CPC [106302.96] and negative control [37852.46]). CHX provided greater reductions of simultaneous levels of Pg [106326.00], Td [3335841] and Tf [61557.47] in the healthy group, as did EO in the diseased groups. CPC provided the greatest reduction [3775319.36] in the periodontitis group. CONCLUSION: Periodontal status influenced the antimicrobial efficacy of preoperative mouthwashes. Therefore, periodontal status should be taken into consideration by clinicians. The antimicrobial efficacy differed among the agents tested. CHX and EO showed the greatest efficacy. The recognition of periodontal condition by clinicians is mandatory to select the most effective preoperative mouthwash.

Clinical and microbiological evaluation of non-surgical periodontal therapy in obese and non-obese individuals with periodontitis: a 9-month prospective longitudinal study.

Objective Obesity is a chronic disease that negatively affects an individual's general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy.

Aggregatibacter actinomycetemcomitans serotypes and JP2 outcomes related to clinical status over 6 years under periodontal maintenance therapy.

OBJECTIVE: This 6-year study evaluatedAggregatibacter actinomycetemcomitans outcomes and their relationship to clinical status. DESIGN: From the eligible individuals (23-70 years of age), 31 regular compliers (between-visit interval < 6 months) were randomly selected and matched for age/sex with 31 irregular compliers (between-visit interval > 6 months). Periodontal clinical examination and subgingival samples were obtained 5 times: T1 (baseline), T2 (after active periodontal therapy), T3 (2 years), T4 (4 years), and T5 (6 years). Total bacteria load, A. actinomycetemcomitans, and red complex species Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola levels were determined by qPCR while PCR was used to determine the occurrence of the a-b-c-d-e-f-g serotypes and the JP2 clone of A. actinomycetemcomitans. Data between groups was compared over time. RESULTS: At baseline PCR revealed A. actinomycetemcomitans prevalence of 9.7 % and JP2 prevalence of 6.7 %. A. actinomycetemcomitans qPCR levels were higher among individuals < 35 years of age and increased at T2 in irregular compliers. At in irregular compliers at the three follow-up visits. Serotypes a, d, and f showed greater values in at least one follow-up visit in regular compliers. A. actinomycetemcomitans showed negative correlation with probing depth (PD) while serotype b showed negative correlations with PD, PI, clinical attachment level and red complex. CONCLUSIONS: Longitudinally, compliance during PMT contributed to lower A. actinomycetemcomitans levels with some degree of correlation with clinical status. However, this study failed to report any positive effect on the occurrence of the most virulent representatives, i.e. serotype b and the JP2 clone.

Clinical and microbiological evaluation of non-surgical periodontal therapy in obese and non-obese individuals with periodontitis: a 9-month prospective longitudinal study

Abstract Objective Obesity is a chronic disease that negatively affects an individual's general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy.

A saliva e seus produtos ­ sua importância na periodontia: revisão da literatura / Saliva and your products ­ your importance in periodontics: literature review

A saliva é um fluido biológico bucal secretado por glândulas salivares e tem grande participação na regulação da microbiota bucal. Atualmente diversas áreas da pesquisa científica adotam o uso da saliva na elaboração de diagnósticos e previsibilidade de terapias afins. Esta tendência se deve ao fato de que o uso da saliva é uma fonte rica de informações moleculares e sua aquisição é relativamente simples por ser uma técnica não invasiva, de fácil coleta, baixo custo, indolor e segura. A saliva pode ser usada no diagnóstico precoce de doenças sistêmicas (câncer, doenças infecciosas e cardiovasculares) e diversas substâncias orgânicas e inorgânicas podem ser detectadas, assim como drogas e seus metabólitos e marcadores biológicos. Particularmente em periodontia, o uso da saliva pode ser importante no diagnóstico, na quantificação de bactérias, na avaliação de interleucinas, nitrato e nitrito entre outros. Assim, o objetivo desta revisão de literatura é atualizar clínicos e periodontistas em relação a importância da saliva como uma ferramenta valiosa no diagnóstico e no monitoramento da condição do hospedeiro em relação às patologias periodontais.(AU)

Saliva is a buccal biological fluid secreted by salivary glands and has great participation in the regulation of the oral microbiota. Currently several areas of scientific research adopt the use of saliva in the elaboration of diagnoses and predictability of related therapies. This trend is due to the fact that the use of saliva is a rich source of molecular information and its acquisition is relatively simple because it is a noninvasive technique, easy to collect, low cost, painless and safe. Saliva can be used for the early diagnosis of systemic diseases (cancer, infectious and cardiovascular diseases) and various organic and inorganic substances can be detected, as well as drugs and their metabolites and biological markers. Particularly in periodontics, the use of saliva may be important in the diagnosis, in the quantification of bacteria, in the evaluation of interleukins, nitrate and nitrite among others. Thus, the objective of this literature review is to update clinicians and periodontists regarding the importance of saliva as a valuable tool in the diagnosis and monitoring of the host condition in relation to periodontal pathologies.(AU)

Effects of chlorhexidine preprocedural rinse on bacteremia in periodontal patients: a randomized clinical trial.

OBJECTIVE: Single dose of systemic antibiotics and short-term use of mouthwashes reduce bacteremia. However, the effects of a single dose of preprocedural rinse are still controversial. This study evaluated, in periodontally diseased patients, the effects of a pre-procedural mouth rinse on induced bacteremia. MATERIAL AND METHODS: Systemically healthy individuals with gingivitis (n=27) or periodontitis (n = 27) were randomly allocated through a sealed envelope system to: 0.12% chlorhexidine pre-procedural rinse (13 gingivitis and 13 periodontitis patients) or no rinse before dental scaling (14 gingivitis and 15 periodontitis patients). Periodontal probing depth, clinical attachment level, plaque, and gingival indices were measured and subgingival samples were collected. Blood samples were collected before dental scaling, 2 and 6 minutes after scaling. Total bacterial load and levels of P. gingivalis were determined in oral and blood samples by real-time polymerase chain reaction, while aerobic and anaerobic counts were determined by culture in blood samples. The primary outcome was the antimicrobial effect of the pre-procedural rinse. Data was compared by Mann-Whitney and Signal tests (p<0.05). RESULTS: In all sampling times, polymerase chain reaction revealed higher blood bacterial levels than culture (p<0.0001), while gingivitis patients presented lower bacterial levels in blood than periodontitis patients (p<0.0001). Individuals who experienced bacteremia showed worse mean clinical attachment level (3.4 mm vs. 1.1 mm) and more subgingival bacteria (p<0.005). The pre-procedural rinse did not reduce induced bacteremia. CONCLUSIONS: Bacteremia was influenced by periodontal parameters. In periodontally diseased patients, pre-procedural rinsing showed a discrete effect on bacteremia control.

Effects of chlorhexidine preprocedural rinse on bacteremia in periodontal patients: a randomized clinical trial

Abstract Objective: Single dose of systemic antibiotics and short-term use of mouthwashes reduce bacteremia. However, the effects of a single dose of preprocedural rinse are still controversial. This study evaluated, in periodontally diseased patients, the effects of a pre-procedural mouth rinse on induced bacteremia. Material and Methods: Systemically healthy individuals with gingivitis (n=27) or periodontitis (n = 27) were randomly allocated through a sealed envelope system to: 0.12% chlorhexidine pre-procedural rinse (13 gingivitis and 13 periodontitis patients) or no rinse before dental scaling (14 gingivitis and 15 periodontitis patients). Periodontal probing depth, clinical attachment level, plaque, and gingival indices were measured and subgingival samples were collected. Blood samples were collected before dental scaling, 2 and 6 minutes after scaling. Total bacterial load and levels of P. gingivalis were determined in oral and blood samples by real-time polymerase chain reaction, while aerobic and anaerobic counts were determined by culture in blood samples. The primary outcome was the antimicrobial effect of the pre-procedural rinse. Data was compared by Mann-Whitney and Signal tests (p<0.05). Results: In all sampling times, polymerase chain reaction revealed higher blood bacterial levels than culture (p<0.0001), while gingivitis patients presented lower bacterial levels in blood than periodontitis patients (p<0.0001). Individuals who experienced bacteremia showed worse mean clinical attachment level (3.4 mm vs. 1.1 mm) and more subgingival bacteria (p<0.005). The pre-procedural rinse did not reduce induced bacteremia. Conclusions: Bacteremia was influenced by periodontal parameters. In periodontally diseased patients, pre-procedural rinsing showed a discrete effect on bacteremia control.

Influência da ciclosporina a nos tecidos periodontais e na osseointegração de implantes: revisão de literatura / Influence of cyclosporine a on periodontal tissue and osseointegration of implants: review of literature

A Ciclosporina A é um potente imunossupressor utilizado no tratamento de diversas patologias mediada imunologicamente. É indicado principalmente no tratamento preventivo da rejeição de órgãos em indivíduos transplantados. Efeitos adversos relacionados com o uso do fármaco como a indução de osteopenia, desequilíbrio no processo de remodelação óssea, desenvolvimento de osteoporose e o aumento gengival são descritos na literatura. O objetivo do presente estudo foi revisar através da literatura os efeitos da ciclosporina A no metabolismo do tecido gengival, ósseo e sua influência como possível fator de risco na osseointegração de implantes. Empregando-se os termos cyclosporine AND dental implants; cyclosporine AND osseointegration; e cyclosporine AND gingival overgrowth como palavras chave, foram levantados artigos na base de dados Pubmed, publicados entre os anos de 2000 a 2016, na língua inglesa e portuguesa. Conclui-se que os efeitos adversos causados pela ciclosporina A podem interferir na saúde bucal dos indivíduos e no sucesso do tratamento odontológico. É fundamental que o cirurgião dentista conheça os mecanismos de ação do medicamento, seus efeitos adversos e interações medicamentosas, a fim de desenvolver estratégias de prevenção e tratamento para usuários do medicamento (AU)

Cyclosporine A is a potent immunosuppressive drug used in the treatment of various immunologically mediated pathologies. It is mainly indicated in the preventive treatment of organ rejection in transplant recipients. Adverse effects associated with using the drug such as the induction of osteopenia, imbalance in bone remodeling, development of osteoporosis and gingival enlargement are described in literature. The aim of this study was to review, through literature, the effects of cyclosporine A in the metabolism of gingival tissue, bone and its influence as a possible risk factor in osseointegration of implants. Using the terms cyclosporine AND dental implants; cyclosporine AND osseointegration; and cyclosporine AND gingival overgrowth as keywords, a search was conducted for articles published in the Pubmed database between the years 2000-2016, in English and Portuguese. It was concluded that the adverse effects caused by cyclosporine A may interfere in the oral health of individuals and the success of the dental treatment. It is essential that the dentist is aware of the action mechanisms of the drug, its side effects and medicinal interactions in order to develop prevention and treatment strategies for users of the drug (AU)

A. actinomycetemcomitans profile and red complex bacterial species of an Afro-Brazilian community: A comparative study.

PURPOSES: The primary aim of this cross-sectional study was to compare the levels of red complex bacteria between Afro-Brazilian and non Afro-Brazilian cohort. The secondary aim was to compare the distribution of both Aggregatibacter actinomycetemcomitans serotype b and its JP2 strains among participants who harboured this bacterial species. METHODS: A total of 84 individuals were included in this study: 42 Afro-descendants (mean age 35.9 ± 13.1 years) and 42 non-Afro-descendants (mean age 36.2 ± 13.1 years) matched (1:1) by periodontal diagnosis, age and gender. All participants received clinical examinations of periodontal pocket depth, clinical attachment level, and plaque and gingival indices. Subgingival samples were taken for microbial analysis. First, genomic DNA (gDNA) was extracted and purified and the quantification of total number of bacterial cells, A. actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola was carried out by qPCR. Then, A. actinomycetemcomitans strains were classified according to serotype b and JP2 profiles by conventional PCR. RESULTS: Clinically, mean PD, mean CAL and percentage of CAL ≥ 3 mm differed between groups (Student's t-test p<0.05). The levels of red complex bacteria between Afro-Brazilian and non-Afro-Brazilian populations were similar. The exception was verified to A. actinomycetemcomitans showing significantly higher levels among Afro-Brazilian descendants in comparison to non-Afro-Brazilian descendants. Afro-Brazilian descendants were clearly infected by more virulent serotype b and JP2 strains. CONCLUSIONS: Despite no statistically significant differences related to the red complex species, Afro-Brazilian descendants harboured higher levels of A. actinomycetemcomitans. Also, our findings confirm that Afro-descendant populations are preferably colonised by A. actinomycetemcomitans serotype b as well as JP2 strains.

Avaliação da eficácia clínica e microbiana de dois protocolos terapêuticos periodontais / Clinical and microbial efficacy of two periodontal therapeutic protocols

O objetivo principal deste ensaio clínico randomizado foi o de avaliar a efetividade clínica e microbiana da terapia de raspagem dental realizada por quadrante em relação a raspagem dental em estágio único. Para responder este objetivo foram incluídos 20 indivíduos com periodontite crônica avançada distribuídos aleatoriamente em 2 grupos a saber: Grupo 1 (10 indivíduos-Raspagem Quadrante), Grupo 2 (10 indivíduos-Raspagem Estágio Único). Mensurações longitudinais foram realizadas durante todo o ensaio confrontando parâmetros clínicos de profundidade de sondagem e nível de inserção clínica para avaliação das terapias. Análise microbiana quantitativa (qPCR) da espécie A. actinomycetemcomitans e P. gingivalis em função do evento “raspagem dental” foi realizada em toda a população do estudo nas etapas pré raspagem [T0] e pós raspagem [T1]. As amostras intrassulculares para a análise do qPCR foram coletadas dos cinco dentes com maior evidência de atividade de doença periodontal ou seja, com as maiores profundidades de sondagem associadas a perda de inserção clínica. Os dados obtidos foram analisados por teste t pareado (p<0.05). Os resultados mostraram que em ambos os grupos houve uma melhora clínica tanto do parâmetro profundidade de sondagem quanto do nível de inserção clinica, já para a frequência dos patógenos testados verificou-se que similarmente ocorreu redução microbiana nos dois grupos investigados. Após análise dos dados conclui-se que ambos protocolos terapêuticos aqui investigados foram eficazes tanto clínica quanto microbiologicamente

The aim of this randomized clinical trial was to evaluate the effectiveness of clinical and microbial effects of scaling and root planing using quadrant debridement or full-mouth debridement. A total of 20 patients diagnosed severe chronic periodontitis were enrolled in this survey: Ten subjects were allocated in quadrant debridement group and 10 subjects in full-mouth debridement group. Longitudinal clinical measurements such as periodontal pocket depth as well as clinical attachment loss were investigated at baseline and at 90 days in both therapies. Also, microbial analysis were performed by qPCR to detect the presence of A. actinomycetemcomitans and P. gingivalis. We collected subgingival samples of 5 severe site from each patient. All clinical and microbial data were analyzed by t student test (p<0.05) and the results showed that both periodontal treatment exhibited good results. Then, we concluded that both treatments provide clinical and microbial benefits for all included patients