Looking for a needle in a haystack: de novo phenotypic target identification reveals Hippo pathway-mediated miR-202 regulation of egg production.

Understanding microRNA (miRNA) functions has been hampered by major difficulties in identifying their biological target(s). Currently, the main limitation is the lack of a suitable strategy to identify biologically relevant targets among a high number of putative targets. Here we provide a proof of concept of successful de novo (i.e. without prior knowledge of its identity) miRNA phenotypic target (i.e. target whose de-repression contributes to the phenotypic outcomes) identification from RNA-seq data. Using the medaka mir-202 knock-out (KO) model in which inactivation leads to a major organism-level reproductive phenotype, including reduced egg production, we introduced novel criteria including limited fold-change in KO and low interindividual variability in gene expression to reduce the list of 2853 putative targets to a short list of 5. We selected tead3b, a member of the evolutionarily-conserved Hippo pathway, known to regulate ovarian functions, due to its remarkably strong and evolutionarily conserved binding affinity for miR-202-5p. Deleting the miR-202-5p binding site in the 3' UTR of tead3b, but not of other Hippo pathway members sav1 and vgll4b, triggered a reduced egg production phenotype. This is one of the few successful examples of de novo functional assignment of a miRNA phenotypic target in vivo in vertebrates.

Predictive factors for obstructive sleep apnea in adults with severe asthma receiving biologics: a single-center cross-sectional study.

PURPOSE: To evaluate the prevalence and potential predictors of obstructive sleep apnea (OSA) in a cohort of adults with severe asthma. METHODS: From March 2021 to December 2021, this cross-sectional study enrolled patients with severe asthma receiving biologics, who were consecutively referred for sleep evaluation irrespective of sleep-related symptoms. Clinical and functional data, including three OSA screening instruments (GOAL, STOP-Bang, and NoSAS) were recorded. All participants underwent a portable sleep test (ApneaLink Air™). OSA diagnosis was based on the respiratory disturbance index ≥ 5.0/h and subclassified according to severity thresholds. Data were subjected to logistic regression tests to identify possible predictors for OSA. Discrimination was estimated from the area under the curve (AUC). RESULTS: Overall, 56 outpatients were included (80% females): 54% with any OSA, 13% with moderate-to-severe OSA, and 4% with severe OSA. In the multivariate analysis, no parameter emerged as an independent predictor for OSA: age (p = 0.080), body mass index (p = 0.060), loud snoring (p = 0.130), and hypertension (p = 0.848). No screening instrument was useful to predict any OSA: GOAL (AUC: 0.714; 95% confidence interval (CI): 0.579-0.849), NoSAS (AUC: 0.645; 95% CI: 0.497-0.793), and STOP-Bang (AUC: 0.640; 95% CI: 0.493-0.788). Similarly, no screening tool was also useful for predicting moderate-to-severe OSA or severe OSA. CONCLUSION: Patients with evere asthma receiving biologics exhibit a high prevalence of OSA. However, no clinical, functional, or OSA screening instrument showed acceptable discriminatory ability to predict the presence of OSA in these patients with severe asthma.

Sleep-Disordered Breathing in Adults with Precapillary Pulmonary Hypertension: Prevalence and Predictors of Nocturnal Hypoxemia.

PURPOSE: To evaluate the frequency of sleep-disordered breathing (SDB) and predictors of the presence of nocturnal desaturation in adults with pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension. METHODS: Outpatients with a hemodynamic diagnosis of precapillary pulmonary hypertension who underwent portable polysomnography were evaluated. Diagnosis and severity of SDB were assessed using three well-established respiratory disturbance index (RDI) thresholds: 5.0/h, 15.0/h, and 30.0/h, while nocturnal hypoxemia was defined by the average oxygen saturation (SpO2) < 90%. Multiple linear regression analysis evaluated the potential relationships among explanatory variables with the dependent variable (average SpO2 values), with comparisons based on the standardized regression coefficient (ß). The R-squared (R2; coefficient of determination) was used to evaluate the goodness-of-fit measure for the linear regression model. RESULTS: Thirty-six adults were evaluated (69.4% females). The majority of the participants (75.0%) had SDB (26 with obstructive sleep apnea [OSA] and one with central sleep apnea [CSA]); while 50% of them had nocturnal hypoxemia. In the linear regression model (R2 = 0.391), the mean pulmonary artery pressure [mPAP] (ß - 0.668; p = 0.030) emerged as the only independent parameter of the average SpO2. CONCLUSION: Our study found that the majority of the participants had some type of SDB with a marked predominance of OSA over CSA, while half of them had nocturnal desaturation. Neither clinical and hemodynamic parameters nor the RDI was a predictor of nocturnal desaturation, except for mPAP measured during a right heart catheterization, which emerged as the only independent and significant predictor of average SpO2.

Long-term respiratory outcomes after COVID-19: a Brazilian cohort study.

Objective: To investigate the prevalence and risk factors for persistent symptoms up to 12 months after hospital discharge in COVID-19 survivors. Methods: This prospective cohort study included patients with COVID-19 discharged from a university hospital in Brazil. Follow-up was performed 2, 6, and 12 months after discharge. Lung function tests and chest computed tomography (CT) were performed 2 months after discharge and were repeated if abnormal. The primary outcomes were the symptoms present, work status, and limitations in daily activities. Results: Eighty-eight patients were included. Dyspnea (54.5%), fatigue (50.0%), myalgia, and muscle weakness (46.6%) were the most common symptoms, which decreased over time. Anxiety was frequent (46.6%) and remained unchanged. One year after discharge, 43.2% of the patients reported limitations in daily activities, and 17.6% had not returned to work. Corticosteroid use was significantly associated with dyspnea and limitations in daily activities. Females had an increased risk of fatigue at the 12-month assessment, with marginal significance after multivariable adjustment. Young age and bronchial wall thickening on admission CT were also risk factors for dyspnea at follow-up. The most common lung function abnormalities were reduced diffusion capacity and small airway disease, which partially improved over time. Conclusions: One year after hospital discharge, more than one-third of patients still had persistent COVID-19-related symptoms, remarkable dyspnea, fatigue, and limitations in daily activities, regardless of acute disease severity. Age, female sex, corticosteroid use during hospitalization, and bronchial thickening on admission CT were associated with an increased risk of sequelae.

Prognostic and clinicopathological significance of podoplanin immunoexpression in oral and oropharyngeal squamous cell carcinoma: A systematic review.

BACKGROUND: Podoplanin (PDPN) is a glycoprotein associated with epithelial-mesenchymal transition, invasion, and metastasis in several types of malignancies, including oral and oropharyngeal squamous cell carcinoma (OSCC). The aim of this systematic review (SR) was to summarize and critically appraise the available evidence about the association between PDPN immunoexpression and clinicopathological features and its utility as a prognostic marker in OSCC. METHODS: Five electronic databases and three gray literature databases were searched for immunohistochemical studies, which were selected in a two-phase process. RESULTS: From 721 records identified, 22 cohort and seven analytical cross-sectional studies were included. Few studies reported that PDPN expression was associated with poorer survival rates in OSCC: overall survival = 4/12, disease-free survival = 4/7, and cancer-specific survival = 2/4 studies. Positive associations were most frequently reported on lymph node involvement, higher histopathological grade, and advanced clinical stages. CONCLUSION: Within limitations of this SR, PDPN may be associated with lymph node involvement, histopathological grade, and clinical stage of OSCC. Current evidence suggests that PDPN could be a useful prognostic marker for OSCC that needs further exploration.

Tissue biomarkers for predicting the risk of oral cancer in patients diagnosed with oral leukoplakia: A systematic review.

OBJECTIVES: We performed a systematic review to evaluate the published biomarkers related to oral leukoplakia (OL), aiming to identify the biomarkers that indicate any future risk of cancer in patients with oral leukoplakia. METHODS: A search strategy was developed for three main electronic databases: PubMed, Cochrane Library, and EBSCO, and also for Google Scholar, until February 28, 2020. The study selection was performed in a two-phase process aiming at studies assessing tissue biomarkers for "malignant transformation of OL." Risk of bias analysis of included studies was performed using the Quality in Prognosis Studies Tool. RESULTS: From 3,130 articles initially identified by searching databases, a total of 46 studies were included in this systematic review, with a combined sample of 3,783 patients, of whom 1,047 presented with malignant transformation of a previously diagnosed OL as reported by the authors. The cancer incidence in the whole group was 27.6% (range: 5.4% to 54.1%). The studies were derived from different geographic areas, including Asia (n = 21), Europe (n = 15), North America (n = 9), and Oceania (n = 1). There were 49 different molecular biomarkers evaluated in the 46 included studies: p53 and podoplanin proteins were the most frequently reported, followed by abnormalities at particular chromosomal loci (e.g., LOH). Risk of bias analysis revealed concerns associated with "measurement of prognostic factor," "study confounding" and "statistical analysis and reporting." CONCLUSIONS: Substantial heterogeneity and lack of standardized reporting of data among the studies were identified. The most promising biomarkers reported to have a significant association with the malignant transformation in OL included podoplanin and chromosomal loci abnormalities. A critical examination of the follow-up studies on OL published so far indicated that tissue biomarkers that could predict the risk of oral cancer in patients with OL are still in a discovery phase.

Malignant transformation of oral submucous fibrosis: A systematic review and meta-analysis.

OBJECTIVES: This systematic review and meta-analysis aimed to determine the proportion of patients who develop oral carcinomas following a diagnosis of oral submucous fibrosis (OSF) in reported longitudinal studies. We also aimed to evaluate the demographic and clinicopathological factors contributing to the progression of OSF to cancer. METHODS: Individual search strategies were applied for the following bibliographic databases: MEDLINE by PubMed, Scopus, Embase, Web of Science, and Grey literature databases until August 30, 2020. Methodological assessment of the risk of bias of the included studies was undertaken using the modified Newcastle-Ottawa scale. Meta-analyses were conducted using a random-effects (DerSimonian and Liard) method to calculate the pooled proportion of the malignant transformation (MT) in OSF patients. RESULTS: Out of 585 records screened, a total of 9 observational studies were included with a total number of 6,337 patients; of these, 292 OSF cases developed carcinomas. The pooled proportion of the MT was 4.2% (95% CI: 2.7%-5.6%) with an annual transformation rate of 0.73%. Subgroup analysis revealed that the pooled MT proportion was significantly higher among population-based studies in comparison with hospital-based ones (p < .005). Most of the studies showed a high risk of bias. In several studies, there was a lack of information about the demographic and clinicopathological characteristics of OSF patients and associated risk indicators; this insufficiency in details hindered the ability to conduct further subgroup analyses. CONCLUSIONS: Despite the poorly reported and the limited number of studies, our analysis confirms that close to 4% of patients diagnosed with OSF may develop oral cancer. Cases with oral epithelial dysplasia had a higher potential for malignant transformation.

Malignant transformation of oral proliferative verrucous leukoplakia: A systematic review and meta-analysis.

OBJECTIVES: To investigate the available evidence on the malignant transformation (MT) of oral proliferative verrucous leukoplakia (PVL). MATERIAL AND METHODS: We searched six main electronic and three grey literature databases in a two-phase process. Cohort studies investigating MT of PVL were eligible for inclusion. The risk of bias (RoB) was assessed using a specific tool developed by the Joanna Briggs Institute. Proportion meta-analyses were performed using a random-effects model. RESULTS: Study selection resulted in the inclusion of 17 studies. The pooled proportion of MT was 43.87% (95% CI = 31.93-56.13). Females (64.02%, 95% CI = 54.87-72.75) were more affected by PVL than males (35.98%, 95% CI = 27.25-45.13). Gingiva (39.6%) and buccal mucosa (21.6%) were the most frequent PVL sites. No conclusive results were found between MT and sex or age distribution, tobacco, or alcohol consumption. Gingiva was the most common site for MT (39.9%), and the most frequent histopathological subtype was conventional squamous cell carcinoma (62.1%). Four studies were classified as low, nine as moderate, and four as high RoB. CONCLUSION: The MT pooled proportion was 43.87%. Among OPMDs, PVL has the highest risk to transform to malignancy. Development and agreement on diagnostic criteria for PVL would reduce the heterogeneity among future studies.

Oral potentially malignant disorders: A consensus report from an international seminar on nomenclature and classification, convened by the WHO Collaborating Centre for Oral Cancer.

Oral potentially malignant disorders (OPMDs) are associated with an increased risk of occurrence of cancers of the lip or oral cavity. This paper presents an updated report on the nomenclature and the classification of OPMDs, based predominantly on their clinical features, following discussions by an expert group at a workshop held by the World Health Organization (WHO) Collaborating Centre for Oral Cancer in the UK. The first workshop held in London in 2005 considered a wide spectrum of disorders under the term "potentially malignant disorders of the oral mucosa" (PMD) (now referred to as oral potentially malignant disorders: OPMD) including leukoplakia, erythroplakia, proliferative verrucous leukoplakia, oral lichen planus, oral submucous fibrosis, palatal lesions in reverse smokers, lupus erythematosus, epidermolysis bullosa, and dyskeratosis congenita. Any new evidence published in the intervening period was considered to make essential changes to the 2007 classification. In the current update, most entities were retained with minor changes to their definition. There is sufficient evidence for an increased risk of oral cancer among patients diagnosed with "oral lichenoid lesions" and among those diagnosed with oral manifestations of 'chronic graft-versus-host disease'. These have now been added to the list of OPMDs. There is, to date, insufficient evidence concerning the malignant potential of chronic hyperplastic candidosis and of oral exophytic verrucous hyperplasia to consider these conditions as OPMDs. Furthermore, due to lack of clear evidence of an OPMD in epidermolysis bullosa this was moved to the category with limited evidence. We recommend the establishment of a global research consortium to further study the natural history of OPMDs based on the classification and nomenclature proposed here. This will require multi-center longitudinal studies with uniform diagnostic criteria to improve the identification and cancer risk stratification of patients with OPMDs, link them to evidence-based interventions, with a goal to facilitate the prevention and management of lip and oral cavity cancer.

Worldwide Effects of Coronavirus Disease Pandemic on Tuberculosis Services, January-April 2020.

Coronavirus disease has disrupted tuberculosis services globally. Data from 33 centers in 16 countries on 5 continents showed that attendance at tuberculosis centers was lower during the first 4 months of the pandemic in 2020 than for the same period in 2019. Resources are needed to ensure tuberculosis care continuity during the pandemic.

Prevalence of mucoepidermoid carcinoma among intraoral minor salivary gland tumors: A systematic review and meta-analysis.

This study aimed to synthetize and critically appraise available evidence regarding mucoepidermoid carcinoma (MEC) prevalence among intraoral minor salivary gland tumors (MiSGT). Five main electronic databases and three grey literature databases were searched. The risk of bias (RoB) was assessed using the Joanna Briggs Institute Critical Appraisal Checklist for Studies Reporting Prevalence Data. Proportion meta-analyses were performed. From 1321 studies identified, 82 were included in qualitative synthesis and 80 in meta-analyses. Sixteen were classified as high, 33 as moderate, and 33 as low RoB. A total of 12 774 MiSGT were found, of which overall MEC prevalence was 16.5% (95% Confidence Interval [95% CI] = 14.8%-18.4%). Most MiSGT were found in the palate (n = 7115), although MEC pooled prevalence in this anatomic location was only 13.6% (95% CI = 11.7%-15.6%). The retromolar area presented the highest pooled prevalence (58.9%; 95% CI = 47.0%-70.3%), followed by gingiva (28.8%; 95% CI = 22.7%-35.4%) and tongue (27.2%; 95% CI = 21.2%-33.6%). Regarding geographic location, Middle East presented the highest pooled prevalence (20.8%; 95% CI = 14.8%-27.6%), followed by America (20.0%, 95% CI = 17.2%-23%) and Europe (15.6%; 95% CI = 9.2%-23.5%). Among MiSGT, the MEC overall prevalence was approximately 16.5%. Although most MiSGT were found in the palate, the retromolar area was proportionally more affected by MEC.

The Role of Cancer-Associated Fibroblasts and Extracellular Vesicles in Tumorigenesis.

Extracellular vesicles (EVs) play a key role in the communication between cancer cells and stromal components of the tumor microenvironment (TME). In this context, cancer cell-derived EVs can regulate the activation of a CAF phenotype in TME cells, which can be mediated by several EV cargos (e.g., miRNA, proteins, mRNA and lncRNAs). On the other hand, CAF-derived EVs can mediate several processes during tumorigenesis, including tumor growth, invasion, metastasis, and therapy resistance. This review aimed to discuss the molecular aspects of EV-based cross-talk between CAFs and cancer cells during tumorigenesis, in addition to assessing the roles of EV cargo in therapy resistance and pre-metastatic niche formation.

Predicting Obstructive Sleep Apnea in Patients with Insomnia: A Comparative Study with Four Screening Instruments.

PURPOSE: Obstructive sleep apnea (OSA) and insomnia are very prevalent disorders, especially in sleep-lab setting, and insomnia may be the presenting complaint of OSA. Here, we aimed to validate No-Apnea as screening tool for OSA in patients with self-reported insomnia complaints and to compare its performance with other models. METHODS: This cross-sectional study involved evaluation of No-Apnea as well as STOP-Bang, NoSAS and Epworth Sleepiness Scale (ESS) in subjects with insomnia being evaluated with full in-lab polysomnography. Discrimination was assessed by area under the curve (AUC), while predictive parameters were calculated by contingency tables. OSA severity was classified based on the apnea/hypopnea index: ≥ 5.0/h as any OSA (OSA≥5), ≥ 15.0/h as moderate/severe OSA (OSA≥15), and ≥ 30.0/h as severe OSA (OSA≥30). RESULTS: Overall, 2591 patients with a clinical diagnosis of insomnia were included. Diagnosis of OSA≥5, OSA≥15, and OSA≥30 was of 76.3%, 53.1%, and 32.6%, respectively. At all levels of OSA severity, No-Apnea had sensitivity ranging from 84.5 to 94.1% and specificity ranging from 58.2 to 35.1%. For screening of OSA≥5, OSA≥15, and OSA≥30, discriminatory ability (AUC) of No-Apnea was: 0.790 [95% confidence interval (CI) 0.770-0.810], 0.758 (95% CI 0.740-0.777), and 0.753 (95% CI 0.734-0.772), respectively. Based on AUCs, No-Apnea, STOP-Bang, and NoSAS performed similar at all levels of OSA severity. The ESS did not present satisfactory discrimination as OSA screening model. CONCLUSIONS: In a large sample of patients with insomnia, No-Apnea, STOP-Bang, and NoSAS, but not ESS, enable satisfactory and similar discrimination at all levels of OSA severity.

Fractional Exhaled Nitric Oxide Measurements and Screening of Obstructive Sleep Apnea in a Sleep-Laboratory Setting: A Cross-Sectional Study.

PURPOSE: Obstructive sleep apnea (OSA) is a common condition characterized by repetitive collapse of the upper airways and intermittent oxygen desaturation, which may lead to airway inflammation. Here, we explored whether fractional exhaled nitric oxide (FeNO) levels provide a non-invasive screening tool of OSA. METHODS: Over a 3-month period, FeNO levels were measured in consecutive non-smoking patients referred for a sleep laboratory. All patients underwent full polysomnography. OSA severity was classified based on the apnea/hypopnea index: ≥ 5.0/h as any OSA, ≥ 15.0/h as moderate/severe OSA, and ≥ 30.0/h as severe OSA. FeNO was measured by a portable device (NIOX-MINO®; Aerocrine AB, Solna, Sweden) and expressed as parts per billion (ppb). Discrimination by area under the curve (AUC) and binary logistic regression were performed. RESULTS: A total of 229 subjects were evaluated. Mean FeNO values were similar among subjects without OSA or with OSA: 16.9 ± 10.6 ppb versus 20.2 ± 14.5 ppb, p = 0.221; respectively. FeNO was not an inclusionary parameter to predict any OSA, moderate/severe OSA, and severe OSA: odds ratio (OR) 1.023 (95% confidence interval [CI]: 0.986-1.062); OR 1.012 (95% CI: 0.991-1.034); and OR 0.999 (95% CI: 0.980-1.018), respectively. The AUC values for FeNO in the diagnosis of any OSA, moderate/severe OSA, and severe OSA showed no discriminatory properties: AUC: 0.567 (95% CI: 0.464-0.670), AUC: 0.541 (95% CI: 0.465-0.618), and AUC: 0.535 (95% CI: 0.459-0.610); respectively. CONCLUSIONS: In a sleep-lab setting, our findings suggest that FeNO measurements are inconsequential in the screening of OSA in adults.

Comparative performance of screening instruments for obstructive sleep apnea in morbidly obese patients referred to a sleep laboratory: a prospective cross-sectional study.

PURPOSE: Obstructive sleep apnea (OSA) is very common occurrence among morbidly obese patients. Our main objectives were to validate the No-Apnea, a 2-item screening tool, in morbidly obese patients and compare its performance with three other instruments: STOP-Bang questionnaire, NoSAS score, and Epworth Sleepiness Scale (ESS). METHODS: A cross-sectional analysis of morbidly obese patients (body mass index [BMI] ≥ 35.0 kg/m2) grouped into two independent samples: bariatric surgery patients (BS) and non-bariatric surgery patients (NBS). All patients underwent overnight polysomnography. Discriminatory ability was assessed by area under the curve (AUC). OSA severity was defined by apnea/hypopnea index cut-off points: ≥ 5.0/h (OSA≥5), ≥ 15.0/h (OSA≥15), and ≥ 30.0/h (OSA≥30). RESULTS: A total of 1017 subjects (40.4% in BS cohort and 59.6% in NBS cohort) were evaluated. In the BS cohort, No-Apnea had similar discrimination to STOP-Bang and NoSAS for predicting OSA≥5 (p = 0.979 and p = 0.358, respectively), OSA≥15 (p = 0.158 and p = 0.399, respectively), and OSA≥30 (p = 0.388 and p = 0.903, respectively). In the NBS cohort, No-Apnea had similar discrimination to STOP-Bang and NoSAS for predicting OSA≥5 (p = 0.528 and p = 0.428, respectively), OSA≥15 (p = 0.825 and p = 0.108, respectively), and OSA≥30 (p = 0.458 and p = 0.186, respectively). Moreover, No-Apnea performed significantly better than ESS in both BS and NBS cohorts (p < 0.001). CONCLUSIONS: No-Apnea is a useful and practical tool for screening of OSA in morbidly obese patients, with non-inferior performance to STOP-Bang questionnaire and NoSAS score.

The synergistic effect of tobacco and alcohol consumption on oral squamous cell carcinoma: a systematic review and meta-analysis.

OBJECTIVES: This systematic review (SR) aimed to summarise and critically appraise available evidence about the association between the synergistic consumption of alcohol and tobacco and the occurrence of oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: Observational studies investigating the association between the synergistic consumption of alcohol and tobacco and OSCC occurrence were included. Studies were selected in a two-phase process and searches were conducted on five main electronic databases, complemented by three grey literature databases. RESULTS: From 3260 records identified, 33 articles were included for qualitative analysis, of which 15 were included in the meta-analyses. Overall, the synergistic consumption was positively associated with the occurrence of OSCC (odds ratio [OR] = 5.37; 95% confidence interval [95%CI] = 3.54-8.14). Increased odds for OSCC occurrence were observed regarding the synergistic consumption of alcohol and smoked tobacco (OR = 4.74; 95%CI = 3.51-6.40), alcohol and smokeless tobacco (OR = 7.78; 95%CI = 2.86-21.14), and alcohol, smoked tobacco, and smokeless tobacco (OR = 16.17; 95%CI = 7.97-32.79). CONCLUSIONS: According to the results of this SR, the synergistic consumption of alcohol and tobacco (both smoked and smokeless) significantly increased the odds for the occurrence of OSCC. CLINICAL RELEVANCE: The knowledge provided by this SR may be useful for a better understanding of differences in the effect of synergistic consumption of alcohol and tobacco products in OSCC development. These data might aid healthcare authorities to develop measures for tobacco and alcohol control considering the needs of their population.

The influence of apical extent of root canal obturation on endodontic therapy outcome: a systematic review.

OBJECTIVES: This systematic review (SR) aimed to investigate the influence of obturation extent on the final outcome of root canal treatment (RCT), by answering the question "among patients requiring RCT on fully formed permanent teeth, is there an association between obturation extent and the final treatment outcome?" MATERIALS AND METHODS: Five electronic databases and three gray literature searches were performed. Observational studies investigating the association between obturation extent and RCT outcome in fully formed permanent teeth with a minimum follow-up of 12 months were included. We evaluated the risk of bias (RoB) in with MAStARI for cohort studies. The overall quality of the evidence was assessed with the GRADE-tool. RESULTS: Twenty-two studies were included, 2 had high RoB, 7 moderate RoB, and 13 low RoB. Underextended obturation demonstrated increased odds of an unfavorable outcome in seven studies, in which the odds varied between 6.94 (95%CI 2.20-21.87) and 1.73 (95%CI 1.02-2.95). Overextended obturation also demonstrated this association in four studies, with odds varying from 1.90 (95%CI 1.23-2.94) to 23.00 (95%CI 5.58-94.75). Due to heterogeneity and the very low level of evidence found in the GRADE analysis, the results from this SR should be interpreted with caution. CONCLUSIONS: Obturation extent seems to influence RCT outcome; overextended and underextended obturations showed higher chance of association with less favorable outcomes than adequate obturation; however, this association was not categorically supported. CLINICAL RELEVANCE: This SR provides information about obturation extent influence on RCT outcome and guides clinicians to make evidence-based decisions during endodontic practice.

Prevalence of oral potentially malignant disorders: A systematic review and meta-analysis.

Oral potentially malignant disorders (OPMD) are chronic conditions, which have a higher risk of transformation to oral squamous cell carcinoma. The aim of this systematic review and meta-analysis was to answer the question: "What is the prevalence of oral potentially malignant disorders among adults?" Studies reporting the prevalence of these conditions (leukoplakia, erythroplakia, oral submucous fibrosis [OSMF], and actinic cheilitis) were selected, only studies in which a clinical assessment and histopathological confirmation were performed were included. Of the 5513 studies, 22 met the inclusion criteria for qualitative and quantitative analyses. The risk of bias (RoB) of the selected studies was assessed using the Joanna Briggs Institute Critical Appraisal Checklist for Studies Reporting Prevalence Data. Seven studies were classified as high risk, 12 as moderate risk, and 3 as low RoB. The meta-analysis showed that the prevalence of OPMD was 4.47% (95% CI = 2.43-7.08). The most prevalent OPMDs were OSMF (4.96%; 95% CI = 2.28-8.62) and leukoplakia (4.11%; 95% CI = 1.98-6.97). OPMDs were identified more commonly in males (59.99%; 95% CI = 41.27-77.30). Asian and South American/Caribbean populations had the highest prevalence rates of 10.54% (95% CI = 4.60-18.55) and 3.93% (95% CI = 2.43-5.77), respectively. The overall prevalence of OPMD worldwide was 4.47%, and males were more frequently affected by these disorders. The prevalence of OPMD differs between populations; therefore, further population-based studies may contribute to the better understanding of these differences.

Whole genome sequence of Mycobacterium kansasii isolates of the genotype 1 from Brazilian patients with pulmonary disease demonstrates considerable heterogeneity.

Mycobacterium kansasii is an opportunistic pathogen and one of the most commonly encountered species in individuals with lung disease. We here report the complete genome sequence of 12 clinical isolates of M. kansasii from patients with pulmonary disease in Brazil.

Application of multiplex real-time polymerase chain reaction assay for simultaneous quantification of Escherichia coli virulence genes in oysters.

Strains of diarrheagenic Escherichia coli (DEC) are involved in foodborne disease outbreaks worldwide, especially the enterohemorrhagic E. coli O157:H7. This study describes two multiplex quantitative real time PCR (qPCR) assays for simultaneous identification and quantification of genes related to virulence of DEC; a triplex reaction for detection and quantification of stxA1, stxA2, and eaeA genes, and a duplex reaction for detection and quantification of eaeA and virA genes. The technique was applied in raw oyster samples for direct quantification of DEC, thereby evaluating the applicability of this methodology for microbiological quality assessment of food. Using custom designed primers and specific MGB probes, a triplex qPCR assay was performed to quantify stxA1, stxA2, and eaeA, and a duplex reaction was performed to quantify virA and eaeA genes. The assays showed high sensitivity, with the detection limit varying between 5 and 17 copies of the genes. The coefficient of determination (R2) of the standard curves was 0.99. The coefficient of variation was < 1% indicated high intra- and inter-assay reproducibilities. The application of this methodology in oyster samples from tropical environment provided direct quantitative data that determined the presence of the genes stxA1 (32.1%), eaeA (28.6%), stxA2 (3.6%), and virA (3.6%). This would prove critical for immediate intervention of control strategies, particularly in oysters that are often ingested as raw food.