Expression of the ErbB2, Ki-67, USP2a and fatty acid synthase (FAS) in oral squamous cell carcinoma

Orientadores: Edgard Graner, Dirce Maria CarraroTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: O sistema ubiquitina (Ub)-proteassomo degrada proteínas intracelulares marcadas com etiquetas de Ub, controlando a disponibilidade destas para a célula. Graner et al. (2004) clonaram recentemente USP2a, uma enzima desubiquitinante que interage com ácido graxo sintase (FAS), provocando sua estabilização e protegendo-a da degradação proteassômica, o que aumenta a meia vida desta enzima metabólica. FAS é a principal enzima envolvida na síntese endógena de ácidos graxos saturados de cadeia longa. Nos tecidos normais a síntese é mínima, pois a maior parte dos ácidos graxos utilizados pelas células é proveniente da dieta. Vários trabalhos recentes têm demonstrado que a expressão de FAS está elevada em diversas neoplasias malignas humanas e, em alguns tumores, sua alta expressão foi associada com pior prognóstico. Há poucos estudos sobre a expressão de FAS em carcinomas espinocelulares (CEC) bucais e todos sugerem a sua participação nas etapas iniciais de transformação maligna. Foi demonstrado que FAS é essencial para a proliferação de linhagens celulares de CECs bucais, tumores estes que expressam maior quantidade desta enzima que o epitélio normal adjacente. Entretanto, pouco se sabe sobre os mecanismos básicos envolvidos no aumento da expressão de FAS em células malignas e seu controle pós-traducional. Recentemente foi demonstrado que ErbB2, um receptor da família ErbB, é capaz de aumentar a expressão de FAS em uma linhagem celular de mama, a qual se torna sensível ao tratamento com bloqueadores da atividade de FAS, entrando em apoptose. Considerando-se o relevante papel biológico de USP2a de proteger FAS da destruição pelo proteassomo e o fato da superexpressão de ErbB2 estar ligada a FAS, o objetivo deste projeto foi estudar a quantidade de RNAs mensageiros provenientes de amostras microdissecadas a laser e suas proteínas em CECs bucais. Nas amostras avaliadas, ErbB2 exibiu dois padrões bastante distintos de marcação, um localizado na membrana plasmática e o outro intra-citoplasmático. Uma alta porcentagem (97,06%) das células fortemente positivas para FAS foi também intensamente marcada para ErbB2 em membrana plasmática, com correlação positiva estatisticamente significante (p = 0,001) e ambos tiveram associação com o marcador de proliferação celular Ki-67. A positividade para ErbB2 e FAS foi correlacionada com os tumores que apresentaram maior espessura de lesão e comprometimento microscópico dos linfonodos. A intensa expressão de ErbB2 na membrana celular ocorreu em áreas de maior diferenciação celular, enquanto que áreas menos diferenciadas e com maior pleomorfismo celular, a expressão citoplasmática de ErbB2 foi mais evidente. As análises realizadas, por meio de qRT-PCR indicaram que os transcritos USP2a, FAS e ErbB2 estão diferencialmente expressos entre as amostras tumorais de CEC bucal quando comparado ao tecido pareado morfologicamente normal das margens adjacentes e ambos estão estatisticamente correlacionadas entre si. Estes resultados mostram que a maioria dos CECs bucais analisados expressa concomitantemente FAS e ErbB2, sugerindo a participação desta última molécula na regulação da expressão gênica de FAS, a qual pode ter um papel biológico na patogênese destas lesões. Além do mais, as proteínas analisadas mostraram ser marcadores moleculares para o prognóstico nestas lesõesAbstract: The ubiquitin (Ub)-proteasome pathway controls cellular protein turnover by degrading targeted intracellular proteins tagged with Ub. Graner et al. (2004) demonstrated that the deubiquitinating enzyme USP2a stabilizes and rescues fatty acid synthase (FAS) from proteassomal degradation. FAS plays a central role in de novo lipogenesis and is down-regulated in most of the tissues, because cells preferentially use circulating dietary fatty acids for the synthesis of new structural lipids. On the other hand, it has been demonstrated that FAS is highly expressed in several human cancers and in some of them its expression is correlated with poor outcome. Few studies describe FAS expression in oral squamous cell carcinoma (SCC), however, all of them suggest that the it is increased at the early stages of malignant transformation. It has been shown that FAS is over-expressed in human oral SCC cell lines and plays an essential role in their proliferation. However, the basic mechanism underlying the control of FAS expression in malignant cells is not known at the moment. It was recently demonstrated that the overexpression of the transmembrane tyrosine kinase receptor ErbB2 is able to increase the expression of FAS in a breast epithelial cell line, which in turn becomes susceptible to apoptosis by FAS inhibition. Considering that USP2a prevents the destruction of FAS through deubiquitination and that ErbB2 overexpression is associated with FAS production, the purpose of the present study was to investigate their mRNA levels by real time PCR in SCC samples and morphologically normal tissue after laser capture microdissection. We also evaluated ErbB2 and FAS protein levels by immunohistochemistry. Two distinct patterns of ErbB2 positivity were identified, a sharply demarcated membrane staining, found in the adjacent normal epithelium and well differentiated areas of the tumors, and a cytoplasmic reaction observed mainly in undifferentiated cells. Most of the lesions (97.06%) that showed a high expression of FAS were also positive for ErbB2 at the cell membrane (p=0.001) and both had correlated with the proliferation marker Ki-67. The immunolabeling for ErbB2 at the cell membrane was also stronger in histologically well differentiated lesions while cytoplasmic positivity was found in undifferentiated tumors. FAS and ErbB2 positivity were associated with microscopic characteristics as thickness and lymphatic embolization of the lesion. Our study also showed a strong positive correlation between ErbB2, FAS and USP2a mRNA expression in the SCC samples compared with normal morphologically tissue of the same source. Taken together, the results presented here show that FAS and ErbB2 are co-expressed in oral SCC and suggest that ErbB2 is able to regulate FAS production in these tumors. Moreover, our data point out that these proteins are significantly associated with a poor prognosisDoutoradoPatologiaDoutor em Estomatopatologi

Objeto Pedagógico: Relatando o Ensino de Arte em Classes Inclusivas

Este estudo investigou os procedimentos didático-pedagógicos para o uso de objetos pedagógicos utilizados pelos professores de artes na grande Florianópolis. Outro aspecto identificado na pesquisa foram os conceitos de inclusão utilizados pelos professores de arte, que fundamentam a prática docente destes profissionais. Na intenção de socializar as análises a partir das instituições visitadas, este artigo sistematiza como se dá a inclusão no espaço escolar e o processo utilizado pelos professores de arte para garantir a participação dos alunos com necessidades especiais

O que dizem os professores de arte sobre as políticas públicas inclusivas

Esse artigo tem como proposta situar as políticas públicas em âmbito nacional, propulsoras da educação inclusiva e seus desdobramentos com relação ao ensino de arte. No processo de entrevista com os professores de artes identificou-se o discurso dos professores de arte acerca das políticas sobre inclusão. O presente texto faz um recorte deste tema buscando as conexões coma formação de professores de artes Essa reflexão objetiva contribuir com o campo da inclusão e da arte-educação, aproximando as duas temáticas e somando-se as discussões sobre o arte-educação

The Impact of Histopathological Features on the Prognosis of Oral Squamous Cell Carcinoma : A Comprehensive Review and Meta-Analysis

ObjectiveOver many decades, studies on histopathological features have not only presented high-level evidence of contribution for treatment directions and prognosis of oral squamous cell carcinoma (OSCC) but also provided inconsistencies, making clinical application difficult. The 8th TNM staging system of OSCC has acknowledged the importance of some histopathological features, by incorporating depth of invasion (DOI) to T category and extranodal extension (ENE) to N category. The aim of this systematic review with meta-analysis is to determine the most clinically relevant histopathological features for risk assessment and treatment planning of OSCC and to elucidate gaps in the literature. MethodsA systematic review was conducted using PRISMA guidelines, and the eligibility criteria were based on population, exposure, comparison, outcome, and study type (PECOS). PubMed, Cochrane, Scopus, and Web of Science were searched for articles exploring the impact of histopathological features on OSCC outcomes with Cox multivariate analysis. Pooled data were subjected to an inverse variance method with random effects or fixed effect model, and the risk of bias was evaluated using quality in prognosis studies (QUIPS). Quality of evidence was assessed with the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) criteria. ResultsThe study included 172 articles published from 1999 to 2021. Meta-analyses confirmed the prognostic potential of DOI, ENE, perineural invasion, lymphovascular invasion, and involvement of the surgical margins and brought promising results for the association of bone invasion, tumor thickness, and pattern of invasion with increased risk for poor survival. Although with a small number of studies, the results also revealed a clinical significance of tumor budding and tumor-stroma ratio on predicted survival of patients with OSCC. Most of the studies were considered with low or moderate risk of bias, and the certainty in evidence varied from very low to high. ConclusionOur results confirm the potential prognostic usefulness of many histopathological features and highlight the promising results of others; however, further studies are advised to apply consistent designs, filling in the literature gaps to the pertinence of histopathological markers for OSCC prognosis. Systematic Review RegistrationInternational Prospective Register of Systematic Reviews (PROSPERO), identifier CRD42020219630.Peer reviewe

Perfil socioeconômico, dietas adotadas e motivações de frequentadores de uma academia em Itapecerica da Serra-SP

A preocupação com o corpo está presente em diferentes gêneros, faixas etárias e classes sociais. O objetivo foi analisar o perfil socioeconômico e as dietas adotadas por praticantes de atividade física, suas motivações e fontes de informação. Trata-se de um estudo descritivo de abordagem quantitativa, com 60 frequentadores de uma academia de ginástica, localizada no município de Itapecerica da Serra - SP. Os dados foram coletados por meio da aplicação de um questionário com 19 questões fechadas. Grande parte do público estudado era do sexo masculino (65,0%), com idade entre 18 a 30 anos (61,7%), solteiros (56,7%), ensino médio completo (48,3%), 43,3% recebiam de um a dois salários mínimos e 35,0% de três a cinco salários. No tocante a dietas, 68,3% relataram está seguindo alguma no momento, sendo que a internet (24,5%) e instrutor da academia (23,4%) eram as principais fontes de informações. Os tipos de dietas mais adotadas foram as dietas hipocalóricas (17,3%) e as indicadas pelo instrutor (16,0%), sendo a saúde (36,0%), o emagrecimento (20%) e a definição do corpo (20%), as principais motivações para sua adesão. Cerca de 38,3% tomam suplementos. As atividades mais praticadas foram musculação (54,8%) e aeróbica (23,1%). Conclui-se que os frequentadores de academia adotam dietas hipocalóricas e outras indicadas pelo instrutor e/ou obtidas em internet, tendo a saúde e a estética como as principais motivações para a adoção de tais planos alimentares. Tornam-se preocupantes as fontes de orientação que este público possui, uma vez que não são especializadas. ABSTRACT Socioeconomic profile, adopted diets and motivations of people who attend a fitness club in the town of Itapecerica da Serra-SPWorries related to bodies are present in different gender, age and social classes. The aim was analysing socioeconomic profile the diets adopted by those who practice physical activities, their motivation and source of information. It is a descriptive study of quantitavive approuch with 60 people who attend a fitness club in the town of Itapecerica da Serra, in the state of São Paulo - Brazil. The data was collected from the answers to a questionary presenting 19 closed questions. A great percentage of the interviewed participants were male (65.0%), aged from 18 to 30 (61.7%), single (56.7%), high school (48.3%), 43.3% gets from one to two minimum salaries a month and 35.0% three to five. A percentage of 68.3% affirms to go on a diet at the moment, being 24.5% internet and 23.4% trainers from the fitness gyms, their main source of information. The diet model most followed was the one suggested by the hipocaloric diet (17.3%) and gym instructors (16.0%); being health (36.0%), loss of weight (20.0%), body definition (20.0%) the main reason to attend the gyms. About 38.3% takes food supplements. The most adopted activity was weightlifting (54.8%) and aerobics (23.1%). Conclusion: The people who attend the fitness gyms go on hipocaloric and others diets indicated by their instructors from the gyms and/or suggested by internet, having health and esthetics as their main motivations to adopt such food programs. The sourses of information these people base themselves become a worry, because it not serious or specialized

Estratégias práticas de ensino sobre insetos para alunos dos anos iniciais do ensino fundamental

Oportunizar um ensino de ciências mais prático e didático possível pode ser mais atrativo e facilitador para o aluno aprender. O artigo objetivou elaborar, desenvolver e validar uma estratégia didática sobre a temática inseto para anos iniciais do ensino fundamental, oportunizando a compreensão das características deste grupo animal; bem como esclarecer a acerca da importância dos insetos na manutenção do equilíbrio do meio. Estas foram desenvolvidas com 165 alunos do primeiro ao quinto ano do ensino fundamental de uma escola pública do município de Palmeira das Missões, RS. Os resultados provenientes de um questionário, foram submetidos à análise de conteúdo e categorização e evidenciaram sobre o que os alunos mais gostaram e aprenderam nas atividades. Os estudantes destacaram os conhecimentos aprendidos sobre as características que identificam os insetos (sejam, as partes do corpo, metamorfose, representantes ou exoesqueleto), seguido da experiência em observar as abelhas, a atividade envolvendo a polinização e o jogo desenvolvido ao ar livre. Os resultados evidenciam que as atividades desenvolvidas foram todas citadas, em maior ou menor quantidade, podendo-se inferir que a sequência didática foi importante e proveitosa, contando com o envolvimento dos alunos, proporcionando um processo de ensino aprendizagem eficaz e prazeroso.&nbsp

Ortho-surgical treatment in skeletal CLII patients with temporomandibular dysfunction: Case report / Tratamento orto-cirurgico em paciente CLII esquelética e com disfunção temporomandibular: Relato de caso

The study reports the ortho-surgical treatment of a patient with class II dentofacial deformity and temporomandibular dysfunction. Patient R.C, 29 years old, mixed race, female, sought the Orthodontics course at Christus University Center, complaining of orofacial pain and reporting other types of pain treatments, but without success. After facial analysis, intraoral and extraoral clinical examination, analysis of panoramic radiography and teleradiographs, it was concluded that in addition to temporomandibular dysfunction the patient had a class II deformity, with anteroposterior deficiency, and occlusal instability. The proposed treatment was ortho-surgical and evaluation of painful symptoms before and after surgery through the TMD Research Diagnostic Criteria questionnaire (RDC / TMD). The patient received pre-surgical orthodontic preparation and was then referred for surgery, in which Le Fort I osteotomy was performed in the maxilla with 3 mm advancement and bilateral sagittal osteotomy in mandibular branches for 5 mm mandibular advancement, in addition to mentoplasty. with a further 5 mm feed. Fixation was performed with the Dual Side 2.0mm System (screws and plates), followed by physical therapy, speech therapy and orthodontic finishing. The patient did not show an improvement in bone positioning, profile and face were more harmonic, without losing its aesthetic identity and there was a significant reduction in pain sensitivity previously presented

Microbiological quality of minimally processed vegetables commercialized in Brasilia, DF, Brazil

This study evaluated the microbiological quality of minimally processed vegetables commercialized in the city of Brasilia, DF, Brazil. A total of 32 samples of different vegetables were purchased from 10 supermarkets. In most samples (78.1%) the populations of psychrotrophic bacteria had a high count ranging from 106 to 108 CFU/g. Thermotolerant coliforms were found in all samples, with populations higher than 102 MPN/g in 15 samples (46.9%). After molecular analyses, E. coli was identified in 16 samples (50.0%) and Salmonella spp. in 4 samples (12.5%). S. aureus was found in 14 samples (43.8%), with counts higher than 103 CFU/g in 4 samples (12.5%). The results obtained in this study showed that 16 samples (50%) were unfit for consumption according to Brazilian legislation. These results indicated the need of adoption of better hygienic practices in the production of minimally processed vegetables to improve quality and microbiological safety

Identification of R-Spondin Gene Signature Predictive of Metastatic Progression in BRAFV600E-Positive Papillary Thyroid Cancer

Papillary thyroid carcinoma (PTC) is the most common malignancy of the thyroid gland and early stages are curable. However, a subset of PTCs shows an unusually aggressive phenotype with extensive lymph node metastasis and higher incidence of locoregional recurrence. In this study, we investigated a large cohort of PTC cases with an unusual aggressive phenotype using a high-throughput RNA sequencing (RNA-Seq) to identify differentially regulated genes associated with metastatic PTC. All metastatic PTC with mutated BRAF (V600E) but not BRAF wild-type expressed an up-regulation of R-Spondin Protein 4 (RSPO4) concomitant with an upregulation of genes involved in focal adhesion and cell-extracellular matrix signaling. Further immunohistochemistry validation confirmed the upregulation of these target genes in metastatic PTC cases. Preclinical studies using established PTC cell lines support that RSPO4 overexpression is associated with BRAF V600E mutation and is a critical upstream event that promote activation of kinases of focal adhesion signaling known to drive cancer cell locomotion and invasion. This finding opens up the potential of co-targeting B-Raf, RSPO and focal adhesion proteins as a pharmacological approach for aggressive BRAF V600E PTC

Trophoblast cell surface antigen 2 expression predicts outcome in oral squamous cell carcinomas

Background Trophoblast cell surface antigen 2 (TROP2) has unclear clinical role in oral squamous cell carcinomas (OSCC). Here, we investigated the association of TROP2 immunoexpression with clinicopathological parameters and survival of OSCC patients. Subjects and Methods Cancer-specific survival (CSS) and disease-free survival (DFS) were assessed in a cohort composed of 266 OSCC. An independent cohort with 88 OSCC samples matched with the normal oral tissue, as well as 17 metastatic lymph nodes, was used for validation. Results Multivariate analysis showed TROP2 as an independent marker of favorable prognosis for both CSS (HR: 0.60, 95% CI: 0.40-0.90, p = .01) and DFS (HR: 0.57, 95% CI: 0.36-0.89, p = .01). Furthermore, TROP2 protein expression was significantly higher in morphologically normal tissues compared to primary tumors (p <.0001) and lymph node metastases (p = .001), and it was significantly associated with CSS (HR: 0.26, 95% CI: 0.09-0.74, p = .008) in the validation cohort. A pooled mRNA analysis performed on the Oncomine (TM) database confirmed the underexpression in OSCC compared with normal tissues (p = .014). Conclusions In summary, our results point to a favorable prognostic significance of TROP2 overexpression in a large cohort of oral cancer patients, suggesting it as an attractive clinical marker.Peer reviewe