FE-SEM Evaluation of Dental Specimens Prepared by Different Methods for In Vitro Contamination

Objective. To evaluate through FE-SEM the cleanliness and dentinal alterations promoted by different methods of dental sample preparation. Methods. Twenty-five human single-rooted teeth were used. The teeth were cleaned and autoclaved in wet medium and randomly divided into 5 groups (n = 5), according to the preparation methods employed—control group: no solutions applied; group 1: cement removal and irrigation with 5.25% NaOCl  + 17 % EDTA for 4 minutes each; group 2: 17%  EDTA + 2.5% NaOCl (4 minutes ultrasonic bath); group 3: cement removal and 17%  EDTA + 5.25%  NaOCl + phosphate buffer solution + distilled water (10 minutes ultrasonic); group 4: 17%  EDTA + 5.25% NaOCl (3 minutes ultrasonic bath). Specimens were analyzed by field emission scanning electron microscope (FE-SEM), at 1500x magnification. Data were submitted to qualitative analysis according to a scoring system and submitted to Kruskal-Wallis test. Results. In ascending order, as to bind parameters, (i) cleanliness: control, group 2, group 3, group 5, and group 4, (ii) dentinal alterations: group 1, group 5, group 2, group 3, and group 4. Conclusion. The proposed protocol was suitable for subsequent microbiological contamination, because it showed less dentinal morphological alterations with increased removal of organic waste

Comparison of rRNA-based reverse transcription PCR and rDNA-based PCR for the detection of streptococci in root canal infections

Objective: The rDNA-based method is unable to distinguish between alive and dead cells. Alternatively, bacterial viability can be assessed by molecular methods based on ribosomal RNA (rRNA). Therefore, this study aimed to detect viable streptococci in root canal samples using rRNA-based reverse transcription polymerase chain reaction (RT-PCR), compared to an rDNA-based PCR assay. Methodology: Microbiological root canal samples were obtained from 32 teeth with primary endodontic infections before (S1) and after chemomechanical preparation (S2), and after removal of intracanal medication (S3). RNA and DNA were extracted, and complementary DNA (cDNA) was synthesized from RNA using RT reaction. cDNA and genomic DNA were subjected to PCR with primers complementary to the 16S rRNA sequences of Streptococcus spp. McNemar’s test was used to compare the detection rate of both assays (P<0.05). Results: Streptococci were detected in 28.12% (9/32) and 37.5% (12/32) of S1 samples using rRNA- and rDNA-based PCR assays, respectively. In contrast, they were detected in only 6.25% (2/32) of S2 samples using rRNA-based RT-PCR, compared to 15.62% (5/32) using rDNA-based PCR. Finally, in S3 samples, streptococci were not detected by rRNA, whereas rDNA-based PCR still detected the bacteria in 12.5% (4/32) of the samples. The total number of PCR-positive reactions in the rDNA-based PCR was higher than in the rRNA-based assay (P<0.05). Conclusions: The rRNA-based RT-PCR showed a lower detection rate of streptococci when compared to the rDNA-based PCR, suggesting that the latter may have detected dead cells of streptococci in root canal samples

A variabilidade da frequência cardíaca está relacionada ao desempenho de resistência em jogadoras de futsal? / Do heart rate variability is relationed to endurance performance in female futsal players?

The study aimed to verify the correlation between resting heart rate variability (HRVrest ) and endurance performance in female futsal players, as well as to evaluate the reliability of this parasympathetic autonomic marker. A total of 16 female futsal players (age: 22 ± 3 years; VO2 max: 42.3 ± 2.0 ml.kg-1.min-1) were evaluated during the first week of preseason training. Vagal modulation was evaluated from the HRVrest (i.e., log-transformed root mean square of successive R-R interval differences-Ln-RMSSD) for two consecutive days, while endurance performance was evaluated by the Yo-Yo Intermittent Recovery Test, Level 1 (Yo-Yo IR1). Pearson correlation was used to analyze the relationship between the variables. Strong correlation between the HRVrest index and endurance performance (r = 0.643; p = 0.007). Reliability was tested through the intraclass correlation coefficient, coefficient of variation (CV), and Bland-Altman analysis of the agreement. Furthermore, acceptable repeatability of HRVrest, but with great inter-subject variability (ICC = 0.670, 95%CI = 0.056-0.885, CV = 15.8%). The current study demonstrated a strong correlation between Ln-RMSSD and endurance performance, and despite the acceptable values of intrasubject reliability,HRVrest presented high inter-individual variability in female futsal players. O objetivo do estudo foi verificar a correlação entre a variabilidade da frequência cardíaca de repouso (VFCRepouso) e o desempenho de resistência em jogadoras de futsal, bem como avaliar a confiabilidade do marcador parassimpático. No total, 16 jogadoras de futsal (idade: 22 ± 3 anos; VO2max: 42.3 ± 2.0 ml.kg-1.min-1) foram avaliadas durante a primeira semana de treinamento da pré-temporada. A modulação vagal foi avaliada a partir da VFC de repouso (isto é, raiz quadrada da média das diferenças sucessivas ao quadrado dos intervalos R-R adjacentes - Ln-RMSSD) por dois dias consecutivos, enquanto o desempenho da resistência foi avaliado pelo Yo-Yo Intermittent Recovery Test, Level 1 (Yo-Yo IR1). A correlação de Pearson foi utilizada para analisar a relação entre as variáveis. A confiabilidade foi testada através do coeficiente de correlação intraclasse, coeficiente de variação e análise de concordância de Bland-Altman. Observou-se uma forte correlação entre o índice de VFCrepouso e o desempenho de endurance (r = 0,643; p = 0,007). Por outro lado, repetibilidade aceitável dos índices de repouso vagal, mas com grande variabilidade interindividual (ICC = 0,670, IC = 0,056-0,885, CV = 15,8%). O presente estudo apresentou forte correlação entre Ln-RMSSD e desempenho de endurance, e mesmo com valores aceitáveis de confiabilidade intrasujeito, a VFC em repouso apresentou alta variabilidade interindividual em jogadoras de futsal

Disinfection of root canals prepared by different techniques of instrumentation and irrigation regimes

O intuito do presente trabalho foi avaliar in vitro a desinfecção de canais radiculares de pré-molares inferiores humanos, variando a técnica de preparo e os regimes de irrigação utilizados. Para isso, 85 espécimes foram padronizados em comprimento e diâmetro apical e, após o devido preparo, receberam contaminação com Enterococcus faecalis e Candida albicans por 28 dias. Os canais foram divididos em 8 grupos: G1 - instrumentação manual associada à irrigação com solução fisiológica; G2 - instrumentação manual associada à irrigação com NaOCl à 1,0%; G3 - instrumentação manual associada à irrigação com NaOCl à 1,0% e AC à 15%; G4 - instrumentação manual associada à irrigação com NaOCl à 5,25%; G5 - instrumentação rotatória associada à irrigação com solução fisiológica; G6 - instrumentação rotatória associada à irrigação com NaOCl à 1,0%; G7 - instrumentação rotatória associada à irrigação com NaOCl à 1,0% e AC à 15%; G8 - instrumentação rotatória associada à irrigação com NaOCl à 5,25%. O grupo controle negativo foi composto por 5 dentes preenchidos com meio de cultura esterilizado. Todos os canais dos grupos experimentais foram preparados até instrumentos com diâmetro final de 0,50mm. Coletas microbianas foram realizada antes e após o PQC, com auxílio de pontas de papel esterilizadas. Raspas de dentina foram coletadas após o PQC com o intuito de verificar a ocorrência de MOs no interior dos túbulos dentinários. Não houve diferença estatística entre as técnicas de instrumentação, quanto à diminuição microbiana no interior dos canais. Quanto aos regimes de irrigação, o soro fisiológico produziu os piores resultados, seguido pelo NaOCl à 1,0%. Já quando acompanhado pelo AC, a desinfecção alcançada não demonstrou diferença estatística quanto aos grupos irrigados com NaOCl à 5,25%.The purpose of this study was to evaluate the in vitro disinfection achieved in root canals of human mandibular premolars, varying the preparation technique and irrigation regimens. For this, 85 specimens were standardized in length and apical diameter, and after proper preparation were contaminated with Enterococcus faecalis and Candida albicans for 28 days. After this period, the canals were divided into eight experimental groups as follows: G1 - hand instrumentation with irrigation with sterile saline, G2 - hand instrumentation with irrigation with NaOCl to 1.0%, G3 - Manual instrumentation with irrigation with NaOCl to 1.0% and citric acid to 15%, G4 - instrumentation associated with irrigation with NaOCl at 5.25%; G5 - rotary instrumentation associated with irrigation with sterile saline and G6 - rotary instrumentation associated with irrigation with NaOCl to 1.0%; G7 - rotary instrumentation with irrigation with NaOCl to 1.0% and citric acid to 15%; G8 - rotary instrumentation with irrigation with NaOCl at 5.25%. Negative control group was composed of 5 teeth had their canals filled with sterile culture medium. All root canals of the specimens of experimental groups were prepared by instruments with a diameter equivalent to the tip end of 0.50 mm. Microbial samplings were performed before and after the PQC with the aid of sterilized paper points. Dentine chips were collected after the PQC in order to verify the occurrence of microorganisms inside the dentinal tubules. The results showed no statistical difference between the techniques of instrumentation, on the microbial reduction. As for systems of irrigation, saline produced the worst results followed by NaOCl 1.0%. When followed by citric acid, disinfection achieved no difference for the groups irrigated with NaOCl at 5.25

Influence of passive ultrasonic irrigation in bacteria and endotoxins from root canals: a randomized clinical study

Este estudo clínico analisou os efeitos dos procedimentos endodônticos e da irrigação ultrassônica passiva (PUI) em bactérias e endotoxinas de canais radiculares. Cinquenta pacientes com dentes com periodontite apical primária foram divididos de forma randomizada em dois grupos: PUI (n=25) e irrigação convencional (IC) (n=25). O preparo químico-cirúrgico (PQC) foi realizado com instrumentos reciprocantes, utilizando-se NaOCl 2,5% durante o preparo; e EDTA 17%, para remoção do magma dentinário. Os canais radiculares foram preenchidos com pasta de hidróxido de cálcio por 14 dias e obturados. Foram realizadas coletas microbiológicas dos canais antes (S1) e após o PQC (S2), após os protocolos de irrigação (S3), após a medicação intracanal (S4) e após a reinstrumentação dos canais (S5). Durante o processamento das amostras, as coletas de 5 casos foram perdidas por fatores diversos. As amostras foram analisadas por PCR quantitativo para detecção e quantificação de bactérias e pelo teste turbidimétrico de LAL para detecção e determinação do nível de endotoxinas. Bactérias e endotoxinas foram observadas em 100% das amostras iniciais coletadas. Em ambos os grupos, houve diminuição significativa na concentração de endotoxinas entre uma etapa do tratamento e a etapa posterior (p0,05). Com relação à redução bacteriana, a PUI foi capaz de reduzir significativamente mais o número de bactérias do que a IC (p 0.05 ). PUI was able to reduce the number of bacteria significantly better than CI (p < 0.05). No significant statistical difference was observed between groups regarding the occurrence of cases wielding positive results for bacteria or endotoxin. It was concluded that PUI was more effective than CI in reducing the number of bacteria but not the amount of endotoxin in the root canal. Furthermore, each step of the endodontic therapy was effective in reducing both the number of bacteria as the amount of endotoxin

Ex vivo evaluation of three instrumentation techniques on E. faecalis biofilm within oval shaped root canals

The objective of the present study was to assess the effectiveness of reciprocating instrumentation in disinfecting oval-shaped root canals infected with Enterococcus faecalis. Forty-five human lower premolars were infected with a culture of E. faecalis (ATCC 29212) for 28 days. Five other teeth that were neither contaminated nor instrumented were used as controls. The 45 specimens were divided into three experimental groups (n = 15) based on the root canal preparation technique used: manual (K-type, Dentsply Maillefer, Ballaigues, Switzerland); rotary (MTwo, VDW GmbH, Munich, Germany); and reciprocating (Reciproc R50, VDW GmbH, Munich, Germany) instruments. During chemomechanical preparation, 21 mL of 2.5% NaOCl was used as the irrigating solution. Microbiological sampling was performed before (S1) and immediately after (S2) the chemomechanical preparation using sterilized paper points. Specimens were then cleaved, and 0.02 g of dentine chips was collected from the root thirds to verify the presence of microorganisms in dentinal tubules. All three preparation techniques reduced the number of microorganisms in the root canal lumen and dentine chips from the root thirds, but no significant differences were observed between the three groups (p > 0.05). Reciprocating instrumentation with Reciproc R50 was effective in reducing the number of microorganisms within the root canal system. Although this technique involves the use of only one file to perform the root canal therapy, it is as effective as conventional rotary instrumentation in reducing theE. faecalis biofilm from the root canal system. However, further clinical investigations are warranted in order to ratify these results

Guidance on mucositis assessment from the MASCC Mucositis Study Group and ISOO: an international Delphi studyResearch in context

Summary: Background: Mucositis is a common and highly impactful side effect of conventional and emerging cancer therapy and thus the subject of intense investigation. Although common practice, mucositis assessment is heterogeneously adopted and poorly guided, impacting evidence synthesis and translation. The Multinational Association of Supportive Care in Cancer (MASCC) Mucositis Study Group (MSG) therefore aimed to establish expert recommendations for how existing mucositis assessment tools should be used, in clinical care and trials contexts, to improve the consistency of mucositis assessment. Methods: This study was conducted over two stages (January 2022–July 2023). The first phase involved a survey to MASCC-MSG members (January 2022–May 2022), capturing current practices, challenges and preferences. These then informed the second phase, in which a set of initial recommendations were prepared and refined using the Delphi method (February 2023–May 2023). Consensus was defined as agreement on a parameter by >80% of respondents. Findings: Seventy-two MASCC-MSG members completed the first phase of the study (37 females, 34 males, mainly oral care specialists). High variability was noted in the use of mucositis assessment tools, with a high reliance on clinician assessment compared to patient reported outcome measures (PROMs, 47% vs 3%, 37% used a combination). The World Health Organization (WHO) and Common Terminology Criteria for Adverse Events (CTCAE) scales were most commonly used to assess mucositis across multiple settings. Initial recommendations were reviewed by experienced MSG members and following two rounds of Delphi survey consensus was achieved in 91 of 100 recommendations. For example, in patients receiving chemotherapy, the recommended tool for clinician assessment in clinical practice is WHO for oral mucositis (89.5% consensus), and WHO or CTCAE for gastrointestinal mucositis (85.7% consensus). The recommended PROM in clinical trials is OMD/WQ for oral mucositis (93.3% consensus), and PRO-CTCAE for gastrointestinal mucositis (83.3% consensus). Interpretation: These new recommendations provide much needed guidance on mucositis assessment and may be applied in both clinical practice and research to streamline comparison and synthesis of global data sets, thus accelerating translation of new knowledge into clinical practice. Funding: No funding was received

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field