Ewing sarcoma is an undifferentiated small-round-cell sarcoma. Although molecular detection of pathognomonic EWSR1-ETS fusions such as EWSR1-FLI1 enables definitive diagnosis, substantial confusion can arise if molecular diagnostics are unavailable. Diagnosis based on the conventional immunohistochemical marker CD99 is unreliable due to its abundant expression in morphological mimics.To identify novel diagnostic immunohistochemical markers for Ewing sarcoma, we performed comparative expression analyses in 768 tumors representing 21 entities including Ewing-like sarcomas, which confirmed that CIC-DUX4-, BCOR-CCNB3-, EWSR1-NFATc2-, and EWSR1-ETS-translocated sarcomas are distinct entities, and revealed that ATP1A1, BCL11B, and GLG1 constitute specific markers for Ewing sarcoma. Their high expression was validated by immunohistochemistry and proved to depend on EWSR1-FLI1-binding to highly active proximal super-enhancers. Automated cut-off-finding and combination-testing in a tissue-microarray comprising 174 samples demonstrated that detection of high BCL11B and/or GLG1 expression is sufficient to reach 96% specificity for Ewing sarcoma. While 88% of tested Ewing-like sarcomas displayed strong CD99-immunoreactivity, none displayed combined strong BCL11B- and GLG1-immunoreactivity.Collectively, we show that ATP1A1, BCL11B, and GLG1 are EWSR1-FLI1 targets, of which BCL11B and GLG1 offer a fast, simple, and cost-efficient way to diagnose Ewing sarcoma by immunohistochemistry. These markers may significantly reduce the number of misdiagnosed patients, and thus improve patient care.

Aim:The aim of the present study was to evaluate the relationship of drinking water fluoride levels with children's intelligence quotient (IQ).Materials and Methods:Water was collected from initially identified endemic fluoride regions according to the geological research of Government of India. Fluoride concentration of the water was assessed by utilizing fluoride ion selective electrode, Orion 9609BN, and categorized on the basis of fluoride concentration into low, medium, and high-fluoride regions, i.e., Virajpet (low fluoride level < 1.2 ppm), Banavara (Medium fluoride level 1.2–2 ppm), and Mastihalli (High fluoride levels > 3 ppm). Government school from all three villages were selected randomly and IQ levels were assessed by using Raven's Standard Progressive Matrices. This test was conducted on each child in the study sample.Results:A significant inverse relationship was found between the fluoride concentration in drinking water and IQ (r value = −0.204; P < 0.000). It was observed that IQ level was negatively correlated with fluoride concentration in drinking water.Conclusion:It is concluded that IQ level was negatively correlated with fluoride level in drinking water. Factors that might affect children's IQ need to be considered, and it is necessary to devise solutions for preventing the harmful effects of excessive intake of fluoride ion to the body.

BackgroundThe Milan system reporting salivary gland cytopathology (MSRSGC) is a tiered classification scheme that is based on risk stratification. The aim of the current study was to assess the risk of malignancy (ROM) and risk of neoplasia (RON) in each of the diagnostic categories proposed by the MSRSGC.MethodsA retrospective analysis and categorization according to the MSRSGC was made of salivary gland fine needle aspirations (FNA) performed from January 2007 to December 2017. The FNA cytology results were correlated with subsequent histological follow‐up.ResultsA total of 578 FNAs were evaluated and histopathology was available for 198 cases (34.2%). The RON and ROM for individual diagnostic categories were: Non‐diagnostic: 52.2% to 13%, non‐neoplastic: 21.4% to 10.7%, atypia of undetermined significance: 74% to 22.2%, benign neoplasm: 100% to 1.1%, salivary gland neoplasm of uncertain malignant potential: 93.3% to 53.3%, suspicious for malignancy (SFM): 100% to 100%, and malignant: 100% to 100%. A diagnosis of 'SFM' or 'malignant' with FNA cytology carried a 100% risk for malignancy, while a diagnosis of “non‐neoplastic,” “benign neoplasm” reduced the probability of malignancy to 3.4%.ConclusionThe MSRSGC is useful for the management of salivary gland lesions as it can successfully differentiate between benign and malignant cases. It will bring uniformity in salivary gland FNA cytology reporting across various institutions globally.

Background: Up to 30-40% of Ewing sarcoma (EwS) patients with non-metastatic disease develop local or metastatic relapse within a time span of 2-10 years. This is in part caused by the absence of prognostic biomarkers that can identify high-risk patients and thus assign them to risk-adapted monitoring and treatment regimens. Since cancer stemness has been associated with tumour relapse and poor patient outcomes, we investigated in the current study the prognostic potential of SOX2 (sex determining region Y box 2)a major transcription factor involved in development and stemnesswhich was previously described to contribute to the undifferentiated phenotype of EwS. Methods: Two independent patient cohorts, one consisting of 189 retrospectively collected EwS tumours with corresponding mRNA expression data (test-cohort) and the other consisting of 141 prospectively collected formalin-fixed and paraffin-embedded resected tumours (validation-cohort), were employed to analyse SOX2 expression levels through DNA microarrays or immunohistochemistry, respectively, and to compare them with clinical parameters and patient outcomes. Two methods were employed to test the validity of the results at both the mRNA and protein levels. Findings: Both cohorts showed that only a subset of EwS patients (16-20%) expressed high SOX2 mRNA or protein levels, which significantly correlated with poor overall survival. Multivariate analyses of our validationcohort revealed that high SOX2 expression represents a major risk-factor for poor survival (HR = 3•19; 95%CI 1•74-5•84; p b 0•01) that is independent from metastasis and other known clinical risk-factors at the time of diagnosis. Univariate analyses demonstrated that SOX2-high expression was correlated with tumour relapse (p = 0•002). The median first relapse was at 14•7 months (range: 3•5-180•7). Interpretation: High SOX2 expression constitutes an independent prognostic biomarker for EwS patients with poor outcomes. This may help to identify patients with localised disease who are at high risk for tumour relapse within the first two years after diagnosis.

Ewing sarcoma is an undifferentiated bone-associated cancer. Although molecular detection of pathognomonic EWSR1-ETS fusions such as EWSR1-FLI1 enables definitive diagnosis, substantial confusion can arise if molecular diagnostics are unavailable. Diagnosis based solely on the conventional immunohistochemical marker CD99 is unreliable due to its abundant expression in morphological mimics. This study aimed to identify novel diagnostic immunohistochemical markers for Ewing sarcoma.We analyzed 768 expression microarrays representing 21 tumor entities including Ewing-like sarcomas to nominate candidate biomarkers. These candidates were validated by immunohistochemistry (IHC) in a tissue microarray (TMA) comprising 174 samples.Microarray, chromatin immunoprecipitation and sequencing (ChIP-Seq) data, and reporter assays were employed to analyze their EWSR1-FLI1-dependency.Our comparative expression analyses revealed that ATP1A1, BCL11B, and GLG1 constitute specific markers for Ewing sarcoma. Analysis of ChIP-Seq and microarray datasets showed that their expression is EWSR1-FLI1-dependent. This outcome corresponded to EWSR1-FLI1-binding to proximal super-enhancers, which showed high activity in reporter assays.Consistently, high ATP1A1, BCL11B, and GLG1 expressions were detected by IHC.Automated cut-off-finding and combination-testing in the TMA demonstrated that detection of high BCL11B and/or GLG1 expression is sufficient to reach 96% specificity for Ewing sarcoma. While 88% of tested Ewing-like sarcomas displayed strong CD99-immunoreactivity, none displayed combined high expression of BCL11B and GLG1.Collectively, we provide evidence that ATP1A1, BCL11B, and GLG1 are EWSR1-FLI1