Comparative study of the expression of p-Akt1, COX-2 and mitochondrial markers in cutaneous and mucosal melanomas

Orientador: Jacks Jorge JuniorTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Melanomas são neoplasias malignas agressivas que ocorrem principalmente na pele. Os melanomas mucosos de cabeça e pescoço, por outro lado, que ocorrem nas mucosas da cavidade bucal e nasal, são neoplasias malignas extremamente agressivas; e devido sua raridade, estudos mais completos sobre possíveis marcadores prognósticos ainda são escassos na literatura. O presente trabalho foi organizado em 5 capítulos que dissertam sobre a expressão de proteína B quinase fosforilada (p-Akt1), ciclooxigenase 2 (COX-2) e marcadores mitocondriais em uma série de melanomas cutâneos e mucosos de cabeça e pescoço. Como principais resultados, podemos destacar, as conclusões de cada capítulo conforme organizado abaixo. No primeiro capítulo, demonstramos que a proteína p-Akt1 teve maior expressão em melanomas mucosos que nos cutâneos e foi identificada como um fator prognóstico independente em melanomas mucosos. No segundo capítulo, foi estudada a expressão de COX-2 em melanomas orais, demonstrando que essa enzima é um fator prognóstico independente para pacientes com estes tumores. No terceiro capítulo, apresentamos uma série de melanomas amelanóticos e foi possível concluir que a apresentação clínica e microscópica desses tumores pode mimetizar outros tipos de lesões, e que, portanto, podem ser mais difíceis de ser diagnosticados. No quarto capítulo, foi comparada a expressão de COX-2 e de Ki67 em melanomas orais amelanóticos e melanomas orais convencionais. Como principal conclusão, a expressão de COX-2 e o índice de proliferação foram maiores em melanomas orais amelanóticos, indicando provavelmente uma maior agressividade destes tumores. No quinto capítulo, foi avaliada a expressão de três marcadores mitocondriais em melanomas cutâneos e mucosos. Como destaque desse último capítulo podemos citar que a quantidade de mitocôndrias nas células tumorais parece desempenhar um papel importante na patogênese dos melanomas. E que, a proteína de fissão 1 (FIS1) pode ser considerada um fator prognóstico em melanomas orais, enquanto a proteína de fusão 2 (mitofusin 2) está associada com pior prognóstico em pacientes com melanomas cutâneos. Em resumo, os cinco capítulos da presente tese contribuíram para o melhor entendimento dos melanomas mucosos e em conjunto apresentam uma série de possíveis marcadores prognósticos que podem ser estudados e indicados como candidatos para terapias-alvoAbstract: Melanomas are tumors with an aggressive behavior that occur mainly in the skin. Mucosal head and neck melanomas, on the other hand occur in the mucous membranes of the oral and nasal cavity. These tumors are extremely aggressive; and due to its rarity, studies with these tumors regarding possible prognostic markers are still scarce in the literature. The present PhD thesis was organized in 5 chapters that discuss the expression of phosphorylated serine/threonine-protein kinase 1 (p-Akt1), cyclooxygenase 2 (COX-2) and mitochondrial markers in a series of cutaneous and mucous melanomas of the head and neck. We have highlighted the main conclusions of each chapter, as organized below. In the first chapter, we demonstrated that p-Akt1 protein was overexpressed in mucosal than in cutaneous melanomas and it was identified as an independent prognostic factor for mucosal melanomas. In the second chapter, we have studied the COX-2 expression in oral melanomas, demonstrating that this enzyme is an independent prognostic factor for patients with these tumors. In the third chapter, we present a series of amelanotic melanomas and the main conclusion was: the clinical and microscopic presentation of these tumors may mimic other types of lesions, and therefore they may be more difficult to diagnose. In the fourth chapter, the expression of COX-2 and Ki67 in amelanotic and conventional oral melanomas was compared. As a main conclusion, COX-2 expression was higher in amelanotic oral melanomas, as well as cell proliferation index in these tumors than in conventional oral melanomas. In the fifth chapter, the expression of three mitochondrial markers in cutaneous and mucosal melanomas was evaluated. We have demonstrated that mucosal melanomas have a higher expression of mitochondrial markers than cutaneous ones. Thus, the high number of mitochondria in tumor cells seems to play an important role in the pathogenesis of melanomas. The fission protein 1 (FIS1) was considered a prognostic factor in oral melanomas, whereas mitofusin 2 is associated with worse prognosis in patients with cutaneous melanomas. In summary, the five chapters of this thesis have contributed to a better understanding of mucosal melanomas and together present a series of possible prognostic markers that can be studied and indicated as candidates for target therapiesDoutoradoPatologiaDoutor em Estomatopatologia2015/25905-1, 2017/16102-8FAPES

Peri-implant peripheral giant cell lesions : report of 13 new cases and comparative histological and immunohistochemical analysis with peripheral and central giant cell lesions

Few cases or peri-implant peripheral giant cell lesions (PGCL) have been reported in the literature. The aim of this study was to report 13 new cases of peri-implant PGCL and compare the expression of smooth muscle actin, Bcl-2 protein, GLUT-1, CD68, osteoprotegerin, receptor activator of nuclear factor kappa-B, Ki-67 and CD34 in these cases with PGCL and central giant cell lesions (CGCL). Clinical data were retrieved from the laboratory records and histological analysis was performed using HE-stained slides. Immunohistochemical reactions for the above mentioned antibodies were performed and digitally scored. Peri-implant PGCL mostly affected the posterior mandible of adult females. CD68 and Bcl-2 expressions were higher in conventional PGCL and CGCL than in peri-implant PGCL (p=0.033 for CD68 and p<.0001 for Bcl-2). Microvessel density was higher in conventional peripheral than in central and peri-implant PGCL (p=0.002). Proliferative index of the mononuclear cells showed no statistically significant differences comparing the three groups but it was higher in peri-implant PGCL. The current study demonstrated that peri-implant PGCL is more common in the posterior mandible of adult females. There were some differences in microvessel density, proliferative activity and expression of CD68 and Bcl-2 among conventional PGCL, peri-implant and CGCL. Further studies are encouraged to better understand these early findings

Comparative expression of cyclooxygenase 2 and Ki67 in amelanotic and conventional oral melanomas

Oral melanomas have some histopathological resemblance with its cutaneous counterpart; however, an aggressive behavior is more common in tumors that occur in the oral cavity. Several markers have been suggested as indicative of tumoral progression and aggressiveness, such as cyclooxygenase 2 (COX-2) and Ki67. In this study, we have compared the expression of COX-2 and Ki67 in a series of amelanotic (n=7) and melanotic oral melanomas (n=22). The cases were selected from 4 pathology laboratories and submitted to the immunohistochemical (IHC) reactions. We analyzed the IHC staining based on a qualitative ? using visual scores; and a computer-assisted method (quantitative) using scanned slides and software for digital analysis. COX-2 was expressed in all oral melanomas; however, its intensity was significantly higher in the amelanotic ones (P<0.001). Similarly, a high Ki67-positivity index was observed in the amelanotic than melanotic ones (P<0.001). Based on these results, we suggest that amelanotic oral melanomas have marked pro-inflammatory and high-proliferative phenotype, justifying their more aggressive behavior compared with the melanotic ones

Comparative effects of fractional radiofrequency and microneedling on the genitalia of postmenopausal women: Histological and clinical changes

Objectives: The authors aimed to evaluate clinical and histological changes induced by Fractional Radiofrequency (FRF) and microneedling in vulvar tissue. Methods: Thirty postmenopausal women were randomly divided into G1 (FRF) and G2 (microneedling) groups. Sub-ablative FRF was executed using disposable fractionated electrodes with an intensity of 8&nbsp;mJ. Microneedling was performed using a derma roller system. The authors evaluated before and after treatment using the Vaginal Laxity Questionnaire (VLQ), EuroQol Five-Dimensional (EQ-5D) questionnaire, and the Blatt and Kupperman Menopausal Index (BKMI). Additionally, the authors performed biopsies of the labia majora for histological analysis pre- and post-treatment. Data were expressed as mean (± standard deviation). A paired t-test was used for intra-group comparison (pre- and post-treatment), with an independent t-test used to compare intergroup data (both pre- and post-treatment). Results: In the G1 group, the VLQ values showed differences compared to the pre-treatment values with the data obtained 60 days after the beginning of the sessions (p&nbsp;=&nbsp;0.01). Similarly, the data changes of the G2 group proved to be significant (p&nbsp;=&nbsp;0.001) across the same time interval. In comparing the groups, VLQ values were not different (p &gt; 0.05). Regarding histological analysis, FRF demonstrated improvement concerning the number of fibroblasts, blood vessels, and fatty degeneration (p &lt; 0.05) compared to the control. Additionally, FRF and microneedling samples showed higher type III collagen and vimentin expression in the immunohistochemical analysis (p &lt; 0.05). Conclusions: The therapies were found to be effective in treating the flaccidity of the female external genitalia. Additionally, histological changes were observed after interventions suggesting collagen remodeling

Epidemiologic analysis of salivary gland tumors over a 10-years period diagnosed in a northeast Brazilian population

Salivary gland tumors (SGT) correspond to a heterogeneous group of lesions with variable biological behavior. The present study aimed to determine the distribution and demographic findings of salivary gland neoplasms in a northeast Brazilian population. A retrospective descriptive cross-sectional study was performed. A total of 588 cases of SGT were diagnosed between 2006 and 2016 of 4 pathology services in the state of Sergipe, Brazil. All cases were reviewed, and data such as sex, age, anatomical location, and histopathological diagnosis were collected. A total of 470 (79.9%) tumors were benign and 118 (20.1%) were malignant. The majority of the patients were females (n=328, 55.8%) with an overall female:male ratio of 1.2:1. The major salivary glands were affected more than the minor glands (69.5% vs. 30.5%). Pleomorphic adenoma (n=419, 71.3%) and mucoepidermoid carcinoma (n=29, 4.9%) were the most frequent benign and malignant tumors, respectively. In addition, both benign and malignant tumors occurred more frequently in the parotid gland (n=300, 51%, p<0.05). The epidemiologic profile and clinical characteristics of SGT were similar to those described in other countries and other regions of Brazil. Epidemiological studies of SGT help to understand their clinical and pathological features and are essential to establish the proper management and prognosis

Mental health and burnout syndrome among postgraduate students in medical and multidisciplinary residencies during the COVID-19 pandemic in Brazil : protocol for a prospective cohort study

Background: The COVID-19 pandemic has led to high levels of physical, psychological, and social stress among health care professionals, including postgraduate students in medical and multidisciplinary residencies. This stress is associated with the intense fear of occupational exposure to SARS-CoV-2, the virus known to cause COVID-19. These professionals are at risk of developing physical and mental illnesses not only due to the infection but also due to prolonged exposure to multidimensional stress and continued work overload. Objective: This study aims to evaluate the prevalence of symptoms suggestive of mental disorders and burnout syndrome and determine the risk factors for burnout among postgraduate students in medical and multidisciplinary residencies in Brazil during the COVID-19 pandemic. Methods: For this prospective cohort study with parallel groups, participants were recruited between July and September 2020 to achieve a sample size of at least 1144 participants. Research instruments such as Depression, Anxiety, and Stress Scale; Patient Health Questionnaire; Brief Resilient Coping Scale; and Oldenburg Burnout Inventory will be used to collect data. Data will be collected in 2 waves: the first wave will include data related to sample characterization and psychosocial evaluation, and the second wave will be launched 12 weeks later and will include an evaluation of the incidence of burnout as well as correlations with the potential predictive factors collected in the first wave. Additionally, we will collect data regarding participants’ withdrawal from work. Results: The recruitment took place from July 29 to September 5, 2020. Data analyses for this phase is already in progress. The second phase of the study is also in progress. The final data collection began on December 1, 2020, and it will be completed by December 31, 2020. Conclusions: We believe the findings of this study will help evaluate the impact of the COVID-19 pandemic on the mental health conditions of health professionals in Brazil as well as contribute to the planning and implementation of appropriate measures that can alleviate these mental health challenges. International Registered Report Identifier (IRRID): DERR1-10.2196/2429

Analyses of COX-2, p-Akt and TGF beta 1 in primary and metastatic melanomas and of the citotoxicity of trans-dehydrocrotonin in melanoma cells

Orientador: Jacks Jorge JuniorDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: O melanoma cutâneo é uma neoplasia maligna que apresenta alta propensão para ocorrência de metástases. Os eventos biológicos mais associados com o processo metastático são: sobrevivência, migração e invasão celular. Diversas vias sinalizadoras induzem esses processos, destacando-se o papel da ciclooxigenase 2 (COX-2) e do Fator de Crescimento Transformante Beta 1 (TGF?-1) que atuam aumentando o metabolismo celular e modulando a interação parênquima-estroma. Além disso, essas vias sinalizadoras têm sido relatadas como cruciais no desenvolvimento de metástases de outras neoplasias malignas. Assim, os objetivos desse trabalho estão divididos em três capítulos, e compreendem: (i) avaliar, por meio de análise imunohistoquímica em Tissue Microarrays (TMAs), a expressão de COX-2, p-Akt e minichromosome maintenance 2 (MCM2) em melanomas primários e metastáticos (MM), correlacionando-os com dados clínicos, histopatológicos e taxas de sobrevida; (ii) avaliar e correlacionar a expressão de TGF?-1 e endoglina (CD105) com a polarização de macrófagos (CD68, CD11c ¿ M1 e CD163 ¿ M2); (iii) analisar, in vitro, o efeito de trans-desidrocrotonina (t-DCTN), um princípio ativo extraído do Croton cajucara Benth, em células de melanoma murino. A sobrevida global dos casos analisados demonstrou correlação positiva com a expressão de COX-2, p-Akt e TGF?-1. Em MM foi observado aumento da expressão de TGF?-1 que apresentou correlação positiva com maior distribuição de macrófagos M2. O número de vasos CD105 positivos também demonstrou correlação positiva com MM e pior prognóstico. Adicionalmente, t-DCTN apresentou efeito citotóxico em todas as concentrações testadas, induzindo apoptose e necrose nas células B16F10 de maneira dose-dependente. Nas células tratadas com t-DCTN, observou-se, por meio de imunofluorescência a diminuição da expressão da metaloproteinase 9. Conclui-se que os biomarcadores COX-2, p-Akt e TGF?-1 apresentam papel biológico relevante na patogênese de MM e na progressão tumoral, destacando-se como potenciais vias específicas para tratamento destas neoplasias. A droga t-DCTN apresentou efeito citotóxico em células B16F10, porém mais estudos são indicados para definir o mecanismo de ação pelo qual essa droga atua, elucidando uma possível opção terapêutica para tratamento de melanomas metastáticos e em fases avançadasAbstract: Cutaneous melanoma is a malignant neoplasm that has a high propensity to develop metastasis. Cell survival, migration, invasion and epithelial-mesenchymal transition are biological events most associated with the metastatic process. Indeed, several signaling pathways act together and culminate with the induction of these processes, highlighting the role of cyclooxygenase 2 (COX-2) and transforming growth factor beta 1 (TGF?-1) which cause increasing cellular metabolism and modulating stromal and tumor cells interactions. Moreover, these signaling pathways have been reported to be crucial for development of metastases in other cancer types. Thus, the present study, divided into three chapters, aimed: (i) to evaluate COX-2, p-Akt1 and minichromosome maintenance 2 (MCM2) expression in primary and metastatic melanomas (MM), and to correlate with clinical, histopathological and survival rates; (ii) to evaluate and correlate TGF?-1 and endoglin (CD105) expression with the polarized macrophages (CD68 ¿ total macrophages, CD11c ¿ M1 macrophages, CD163 ¿ M2 macrophages); (iii) analyze in vitro the effect of trans-dehydrocrotonin (t-DCTN), a drug extracted from Croton cajucara Benth in murine melanoma cells (B16F10). We demonstrated that TGF?-1, CD105, COX-2, p-Akt, CD163, Ki67 and MCM2 are associated with lower specific-disease survival in primary and metastatic tumors, supporting their role in melanoma metastatic progression. Overexpression of these biomarkers and higher number of vessels CD105 positive also showed a positive correlation with MM and worse prognosis. Additionally, t-DCTN showed cytotoxic effect in all concentrations tested, induced apoptosis and necrosis in the B16F10 cells in a dose-dependent manner. In cells treated with t-DCTN, it was observed by immunofluorescence a reduction in the expression of metalloproteinase 9 (MMP-9). We conclude that the biomarkers (COX-2, p-Akt1 and TGF?-1) have important biological role in the pathogenesis of MM, and tumor progression, especially as potential specific targets for metastatic melanomas treatment. Furthermore, t-DCTN drug showed cytotoxic effect on B16F10 cells (metastatic) murine melanoma, however further studies are indicated to define the mechanism of action of this drug, elucidating a possible more effective and less toxic therapeutic option for treatment of advanced stages and metastatic melanomasMestradoPatologiaMestre em EstomatopatologiaCAPE