miR-22 and miR-205 Drive Tumor Aggressiveness of Mucoepidermoid Carcinomas of Salivary Glands.

OBJECTIVES: To integrate mRNA and miRNA expression profiles of mucoepidermoid carcinomas (MECs) and normal salivary gland (NSGs) tissue samples and identify potential drivers. MATERIAL AND METHODS: Gene and miRNA expression arrays were performed in 35 MECs and six NSGs. RESULTS: We found 46 differentially expressed (DE) miRNAs and 3,162 DE mRNAs. Supervised hierarchical clustering analysis of the DE transcripts revealed two clusters in both miRNA and mRNA profiles, which distinguished MEC from NSG samples. The integrative miRNA-mRNA analysis revealed a network comprising 696 negatively correlated interactions (44 miRNAs and 444 mRNAs) involving cell signaling, cell cycle, and cancer-related pathways. Increased expression levels of miR-205-5p and miR-224-5p and decreased expression levels of miR-139-3p, miR-145-3p, miR-148a-3p, miR-186-5p, miR-338-3p, miR-363-3p, and miR-4324 were significantly related to worse overall survival in MEC patients. Two overexpressed miRNAs in MEC (miR-22 and miR-205) were selected for inhibition by the CRISPR-Cas9 method. Cell viability, migration, and invasion assays were performed using an intermediate grade MEC cell line. Knockout of miR-205 reduced cell viability and enhanced ZEB2 expression, while miR-22 knockout reduced cell migration and invasion and enhanced ESR1 expression. Our results indicate a distinct transcriptomic profile of MEC compared to NSG, and the integrative analysis highlighted miRNA-mRNA interactions involving cancer-related pathways, including PTEN and PI3K/AKT. CONCLUSION: The in vitro functional studies revealed that miR-22 and miR-205 deficiencies reduced the viability, migration, and invasion of the MEC cells suggesting they are potential oncogenic drivers in MEC.

Vaccine confidence and hesitancy in Brazil.

Despite the overall benefits of immunization, vaccine hesitancy has been a growing trend and has been associated with the resurgence of vaccine-preventable diseases. The aim of this study was to assess vaccine confidence and hesitancy in Brazil, as part of a wider project to map vaccine confidence globally. One thousand subjects were interviewed, either online or face-to-face, based on a general questionnaire regarding perceptions on vaccines and vaccination. Further exploratory questions were used with the subset of respondents who were parents of children aged under 5. Such questions extracted information regarding vaccination behavior, opinions on vaccination and government health services, and vaccine hesitancy. Reasons for hesitancy were classified as relating to confidence, convenience and/or complacency, and the population was also analyzed socio-demographically. The results showed that overall confidence in immunization was higher than confidence in family planning services, community health workers and emergency services. Seventy-six people reported hesitancy to vaccinate. The commonest reasons for hesitancy were issues with confidence (41.4%), efficacy/safety of the vaccine (25.5%) and concerns about adverse events (23.6%). The sociodemographic analysis revealed that vaccine hesitancy was associated with marital status, level of education and income. Despite overall vaccine confidence being high, a clear trend toward lower levels of confidence was associated with higher levels of hesitancy, which warrants on-going monitoring, due to the dynamic and changing nature of vaccine hesitancy.

Vaccine confidence and hesitancy in Brazil / Confiança nas vacinas e hesitação em vacinar no Brasil / Confianza y renuencia a las vacunas en Brasil

Despite the overall benefits of immunization, vaccine hesitancy has been a growing trend and has been associated with the resurgence of vaccine-preventable diseases. The aim of this study was to assess vaccine confidence and hesitancy in Brazil, as part of a wider project to map vaccine confidence globally. One thousand subjects were interviewed, either online or face-to-face, based on a general questionnaire regarding perceptions on vaccines and vaccination. Further exploratory questions were used with the subset of respondents who were parents of children aged under 5. Such questions extracted information regarding vaccination behavior, opinions on vaccination and government health services, and vaccine hesitancy. Reasons for hesitancy were classified as relating to confidence, convenience and/or complacency, and the population was also analyzed socio-demographically. The results showed that overall confidence in immunization was higher than confidence in family planning services, community health workers and emergency services. Seventy-six people reported hesitancy to vaccinate. The commonest reasons for hesitancy were issues with confidence (41.4%), efficacy/safety of the vaccine (25.5%) and concerns about adverse events (23.6%). The sociodemographic analysis revealed that vaccine hesitancy was associated with marital status, level of education and income. Despite overall vaccine confidence being high, a clear trend toward lower levels of confidence was associated with higher levels of hesitancy, which warrants on-going monitoring, due to the dynamic and changing nature of vaccine hesitancy.

Apesar dos benefícios globais da imunização, a hesitação em vacinar é uma tendência crescente que tem sido associada ao ressurgimento das doenças imunopreveníveis. O estudo teve como objetivo avaliar a confiança nas vacinas e a hesitação em vacinar no Brasil, como parte de um projeto mais amplo para mapear a confiança em vacinas em nível global. Foram entrevistadas mil pessoas, direta ou virtualmente, usando um questionário geral sobre percepção em relação às vacinas e à vacinação. Foram utilizadas perguntas exploratórias adicionais no subconjunto de entrevistados que eram pais de crianças abaixo de cinco anos de idade. Essas perguntas produziram informações sobre o comportamento em relação à vacinação, opiniões sobre vacinação e serviços públicos de saúde e hesitação em vacinar. Os motivos da hesitação foram classificados em relação à confiança, conveniência e/ou acomodação, e a população também foi analisada conforme as características sociodemográficas. Os resultados mostraram que a confiança geral na imunização foi maior do que nos serviços de planejamento familiar, agentes comunitários de saúde e serviços de emergência. Setenta e seis pessoas relataram hesitação em vacinar. Os motivos mais frequentes da hesitação diziam respeito a confiança (41,4%), eficácia/segurança da vacina (25,5%) e preocupações com eventos adversos (23,6%). A análise sociodemográfica mostrou que a hesitação em vacinar estava associada ao estado civil, escolaridade e renda. Apesar da alta confiança geral na vacinação, uma clara tendência para níveis de confiança mais baixos esteve associada a níveis mais altos de hesitação, o que justifica o monitoramento permanente dessa tendência, em função da natureza dinâmica da hesitação em vacinar.

A pesar de lo beneficios generales de la inmunización, la renuencia a la vacunación ha sido una tendencia en crecimiento que ha sido asociada con el resurgimiento de las enfermedades prevenibles por vacunación. El objetivo de este estudio fue evaluar la confianza y renuencia a las vacunas en Brasil, como parte de un proyecto más amplio para mapear la confianza en las vacunas globalmente. Mil sujetos fueron entrevistados, bien en línea o cara-a-cara, mediante un cuestionario general respecto a sus percepciones sobre las vacunas y la vacunación. Se utilizaron otras preguntas exploratorias con el subconjunto de encuestados, que eran padres de niños con una edad inferior a los cinco años. Tales preguntas recabaron información respecto al comportamiento sobre la vacunación, opiniones sobre vacunación y servicios de salud gubernamentales, así como su renuencia a las vacunas. Las razones para esta última se clasificaron como aquellas relacionadas con la confianza, conveniencia y/o complacencia, y la población fue también analizada sociodemográficamente. Los resultados mostraron que la confianza general en inmunización fue más alta que la confianza en servicios de planificación familiar, trabajadores de salud comunitarios y servicios de emergencia. Setenta y seis personas informaron de renuencia a la vacunación. Las razones más comunes para la renuencia fueron temas relacionados con la confianza (41,4%), eficacia/seguridad de la vacuna (25,5%) y preocupaciones sobre efectos adversos (23,6%). El análisis sociodemográfico reveló que la renuencia a la vacunación estaba asociada con el estado civil, nivel de educación e ingresos. A pesar de que la confianza general en las vacunas es alta, existe una clara tendencia hacia niveles más bajos de confianza, que estaba asociada con altos niveles de renuencia, lo que garantiza una supervisión permanente, debido a la dinámica y naturaleza cambiante del rechazo a las vacunas.

15d-PGJ2 as an endoplasmic reticulum stress manipulator in multiple myeloma in vitro and in vivo.

Multiple myeloma (MM) is characterised by intense protein folding and, consequently endoplasmic reticulum (ER) stress. The prostaglandin 15d-PGJ2 is able to raise oxidative stress levels within the cell and potentially trigger cell death. The aim of this study was to evaluate the antineoplastic effect of 15d-PGJ2 on MM in vitro and in vivo via ER and oxidative stress pathways. MM.1R and MM.1S cell lines were treated with 15d-PGJ2 at 1-10µM and evaluated with regard to proliferation, mRNA expression of PRDX1, PRDX4, GRP78, GRP94, CHOP, BCL-2 and BAX. Stress data was validated via oxidized glutathione assays. MM.1R cells were inoculated into NOD/SCID mice, which were subsequently treated daily with 15d-PGJ2 at 4mg/kg or vehicle (control), with tumour volume being monitored for 14days. 15d-PGJ2 reduced cell proliferation, induced cell death and apoptosis at 5µM and 10µM and Stress-related genes were upregulated at the same doses. Oxidized glutathione levels were also increased. 15d-PGJ2 at 4mg/kg in vivo halted tumour growth. In conclusion, 15d-PGJ2 induced myeloma cell death via ER stress in vitro. 15d-PGJ2 in vivo also inhibited tumour growth.

Mammaglobin and DOG-1 expression in polymorphous low-grade adenocarcinoma: an appraisal of its origin and morphology.

BACKGROUND: Polymorphous low-grade adenocarcinoma (PLGA) remains a diagnostic challenge for most pathologists due to its large spectrum of histological patterns. In this study, the expression of two new markers recently described for salivary gland tumors was studied in PLGA. METHODS: The morphology of 33 cases of PLGA was carefully evaluated using hematoxylin-and-eosin-stained sections and confirmed by immunohistochemistry for cytokeratin 7, vimentin, and S-100. Periodic acid-Schiff with diastase digestion was also used. The expression of mammaglobin and DOG-1 was carried out using the EnVision System. Mammaglobin was assessed according to the percentage of positively stained tumor cells, while DOG-1 was evaluated according to its presence and site. For MCM-2 and Ki-67, markers of proliferation, the labeling index of cell nuclei positivity was evaluated using total cell number. The ETV6-NTRK3 fusion was examined by fluorescence in situ hybridization analysis. RESULTS: The histological patterns of the tumor were classified as lobular or non-lobular. For the non-lobular pattern, tubular, cribriform, glomeruliform, trabecular, and papillary patterns were observed. Mammaglobin was present in all PLGA cases, and its expression was stronger (P = 0.01) in the lobular than in the non-lobular pattern. The expression of DOG-1 was present in the apical portion and cytoplasm of the cells. Proliferation markers were low for all cases independent of histological pattern. CONCLUSIONS: Polymorphous low-grade adenocarcinoma has been confirmed to originate from the intercalated duct and to feature high expression of mammaglobin in its lobular pattern resembling that of mammary secretory analogue carcinoma, except for the ETV6 gene rearrangement.

Goldenhar Syndrome in a pediatric patient: a case report and review of literature / Síndrome de Goldenhar em um paciente infantil: relato de caso e revisão da literatura

ABSTRACT Goldenhar Syndrome is a rare, generally sporadic condition, whose physical manifestations include asymmetric incomplete facial development, epibulbar dermoids, auricular malformations e auricular appendices, vertebral anomalies, central nervous system disturbances, ocular irregularities and visceral abnormalities. Its etiology is not fully understood, with its incidence ranging from 1:3500 to 1:5600 live births, and a gender ratio of 3:2 (male: female), presenting more frequently in children with congenital deafness. The purpose of this case report was to describe clinical presentation of Goldenhar syndrome in a five-year old male with no significant family history, where almost all of the classic signs of the condition were present. Considering its involvement in craniofacial structures, its rarity, and its wide spectrum of symptoms and associated systemic abnormalities, knowledge of this condition is paramount for the dental surgeon.

RESUMO A Síndrome de Goldenhar é rara e suas principais manifestações físicas incluem assimetria e desenvolvimento facial incompleto, tumores dermoides epibulbares, malformações na orelha e apêndices auriculares, anomalias vertebrais, distúrbios no sistema nervoso central, irregularidades oculares e anomalias viscerais. A etiologia desta condição não é claramente esclarecida e apresenta-se geneticamente variável. A incidência da Síndrome de Goldenhar pode variar de 1:3500 até 1:5600 nascidos vivos e relação por sexo de 3:2 (masculino: feminino), apresentando mais frequentemente em crianças com surdez congênita. O objetivo deste trabalho é apresentar um caso clínico de Síndrome de Goldenhar em paciente infantil de 5 anos de idade do sexo masculino, sem história familiar relevante relacionada à síndrome e que apresenta características claras desta condição. Considerando o envolvimento das estruturas craniofaciais da Síndrome de Goldenhar, sua raridade e o amplo espectro de sintomas e anormalidades sistêmicas associadas, o conhecimento desta condição é primordial para o cirurgião-dentista.

Peripheral giant cell granuloma associated with a dental implant: a case report and review of the literature.

The peripheral giant cell granuloma (PGCG) is a nonneoplastic lesion commonly caused by local irritation. This report describes a 46-year-old Caucasian male who presented with a PGCG associated with a dental implant. The dental implant was originally placed in August 2012. Ten months later, the patient presented with a well-circumscribed lesion associated with and covering the implant, at which time the lesion was excised. Four months later, due to recurrence of the lesion, a deeper and wider excisional biopsy with curettage of the adjacent bone was performed. No evidence of recurrence has been reported after 12 months of follow-up. Immunohistochemistry, using the antibody CD68, was performed to investigate the origin of the multinucleated giant cells, with their immunophenotype being similar to those of other giant cell lesions, including central giant cell granuloma, foreign-body reactions, and granulomatous reactions to infectious agents.

The EGF signaling pathway influences cell migration and the secretion of metalloproteinases by myoepithelial cells in pleomorphic adenoma.

During tumor development, benign neoplastic cells are influenced by the expression of cytokines, growth factors, and proteases present in the tumor microenvironment. Epidermal growth factor (EGF) is the most studied growth factor and is considered important for cell proliferation and migration. Metalloproteinases (MMPs) are also involved in tumor progression. The present study aimed to analyze the proliferation, viability and migration index of pleomorphic adenoma myoepithelial cells, in addition to the secretion of MMPs with EGF supplementation. Benign myoepithelial cells were cultured with two different EGF doses (5 and 10 ng/ml), and the influence of EGF on cell proliferation and viability, using trypan blue and MTT assays, respectively, after 24, 48, and 72 h, was evaluated. To analyze cellular morphology, hematoxylin-eosin staining and indirect immunofluorescence using the anti-vimentin antibody, was performed. In vitro migration assays were performed in Transwell chambers with an 8-µm pore covered with Matrigel and supplemented with 5 or 10 ng/ml of EGF, after 96 h. After 4 days of cell culture, ELISA was performed to determine the MMP-2 and MMP-13 levels. One-way analysis of variance (ANOVA) with post hoc Tukey test was applied, with a significance level of 0.05. The results revealed that EGF influences myoepithelial cell morphology, without alteration of proliferation and viability. The migration assay showed that EGF increased the mean index from 16 % in the control group to 40 and 76 % for 5 and 10 ng/ml of EGF, respectively. ELISA revealed that when the cells were supplemented with either of the EGF doses, an increase in MMP-2 levels was observed when compared with the control group (C). This study concludes that EGF aids in the production of MMP-2, which favors the dissolution of the basement membrane, contributing to cell migration and tumor progression, hence permitting contact between the myoepithelial cells and stroma.