Epithelial ovarian cancer is the most lethal gynecologic malignancy, with a 5-year survival rate of less than 40%. This is due in part to a lack of early detection markers and lack of specific symptoms during early disease. The laying hen is the only accessible animal model which develops epithelial ovarian cancer spontaneously, with features closely resembling the human disease. It has been estimated that approximately 30% of all cancers can be prevented with diet, exercise, and maintenance of an optimal weight, and the chronic low-grade inflammation that accompanies obesity is implicated as a causal factor in the development of cancer. Flaxseed, a rich plant source of anti-inflammatory omega-3 fatty acids and lignans which act as phytoestrogens and antioxidants, exhibits chemosuppressive effects against the development and progression of ovarian cancer. We have shown that a diet of 10% flaxseed reduces the incidence and severity of ovarian cancer when fed to laying hens over 4 years, due in part to the ability of flaxseed to suppress the production of proinflammatory PGE2 in the ovary by decreasing expression of COX enzymes. To investigate other potential specific mechanisms by which flaxseed acts to suppress ovarian cancer, we examined expression and activity of pathways known to be involved in the etiology and progression of human epithelial ovarian cancer in ovarian cancer in the laying hen, and determined whether flaxseed affected these pathways during cancer development. We investigated the effect of flaxseed and its individual components upon oxidative stress in the normal ovary and in ovarian cancer by analyzing expression of target genes of the NRF2 transcription factor. The NRF2 pathway is a "master switch" that regulates expression of ROS-responsive detoxification genes. Results revealed that expression of four genes was significantly downregulated in then ovaries of hens on the defatted flaxmeal (DFM) and whole flaxseed (WF) diets compared to hens on diets that are high in pro-inflammatory omega-6 fatty acids, suggesting that flaxseed decreases oxidative stress in the ovary. Conversely, one target gene was upregulated in ovarian cancer compared to normal ovaries, and this observation was not affected by flaxseed. Additionally, nuclear accumulation Nrf2 protein was not observed in tumor cells, suggesting that flaxseed does not exert chemosuppressive effects by modulating NRF2 signaling in ovarian cancer. To further investigate pathways potentially regulated by flaxseed, we performed a microarray with 44k features and found that a set of genes involved in branching morphogenesis was upregulated in ovarian cancer and significantly decreased by flaxseed, including E-cadherin and miR-200, suggesting that flaxseed impedes the activity of an aberrantly activated developmental program that controls gland formation during ovarian cancer progression. Lack of nuclear accumulation of ZEB1 protein in tumor cells suggests that this decrease in expression is likely not due to EMT. Finally, due to its known roles in controlling developmental programs such as EMT as well as regulating cell growth and proliferation, we performed a set of experiments to examine activity of the TGF-beta pathway. PCR array analysis revealed that SMAD target genes, ligands, receptors, and co-regulatory proteins were upregulated in ovarian tumors from hens on both diet groups, suggesting TGF-beta signaling is enhanced in ovarian cancer. However, expression of SMAD6 and SMAD7 was upregulated in tumors from hens on the flaxseed diet but not control diet, with SMAD7 protein being expressed in both epithelial tumor cells and intratumoral stromal cells. Additionally, immunohistochemical staining for pSMAD2/3 was decreased in epithelial tumor cells and absent from intratumoral stromal cells in tumors from hens on the flaxseed diet compared to tumors from hens on the control diet, and these data together suggest that flaxseed may inhibit pro-oncogenic TGF-beta signaling in ovarian cancer. Finally, flaxseed prevents the downregulation of expression of p15 and the upregulation of CCNA and CCNE in ovarian tumors, suggesting that flaxseed may slow cell cycle progression. Data from these studies provides preliminary evidence that flaxseed exerts pleiotropic effects upon gene expression to negatively regulate pathways driving the progression of ovarian cancer, including aberrant TGF-beta signaling and glandular development. These studies provide groundwork for in vitro studies to test the specific effects of flaxseed upon proteins involved in TGF-beta signaling and upon the expansion of tumor epithelia.

Resumo: O câncer de ovário apresenta diagnóstico tardio e alta letalidade, devido à falta de biomarcadores sensíveis e específicos e à rápida progressão desse câncer, assintomático em estadios iniciais. As células imunes têm a capacidade de eliminar as células malignas e regular a progressão tumoral. Contudo, as células imunes do microambiente tumoral são disfuncionais e falham no controle da expansão tumoral podendo, inclusive, promover o crescimento da neoplasia. Apesar das inúmeras tentativas de se correlacionar o grau e o tipo de infiltrado celular com o prognóstico ou sobrevida do paciente com câncer de ovário, não há consenso sobre o real significado do infiltrado leucocitário nesses casos. Desse modo, este estudo tem como objetivo a ampliação dos conhecimentos relativos à imunidade inata em mulheres com câncer de ovário, através caracterização dos aspectos fenotípicos celulares da imunidade inata sérica. Trata-se de estudo transversal onde foram avaliadas 36 mulheres submetidas a exame clínico, ginecológico, ultrassonografia transvaginal e tratamento com laparotomia nos casos indicados de massa pélvica. De acordo com o resultado desses exames, as mulheres selecionadas foram agrupadas nos grupos: controle- ausência de neoplasia, com neoplasia ovariana benigna e neoplasia ovariana maligna. Realizou-se dosagem sérica de moléculas de expressão de superfície de células da resposta imune inata com análise através da citometria de fluxo. As diferenças entre os grupos foram avaliadas pelo teste de Mann-Whitney (dois grupos) ou Kruskal-Walis (três grupos) conforme indicados. As diferenças com valor de p<0,05 foram consideradas significativas. Foram selecionadas 36 pacientes: 10 mulheres no grupo controle, 9 no grupo de neoplasia ovariana benigna e 17 no grupo de neoplasia maligna. Mais de 70% das pacientes com câncer de ovário apresentavam-se com doença... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: Ovarian cancer presents late diagnosis and high mortality due to lack of sensitive and specific biomarkers and the rapid progression of this cancer, asymptomatic in early stages. Immune cells have the ability to eliminate malignant cells and regulate tumor progression. However, the immune cells of the tumor microenvironment are dysfunctional and fail to control the tumor growth and may even promote the growth of cancer. Despite numerous attempts to correlate the degree and type of cellular infiltrate and the prognosis or survival of patients with ovarian cancer, there is no consensus about the real meaning of the leukocyte infiltrate in these cases. Thus, this study aims to increase knowledge about the innate immunity in women with ovarian cancer through characterization of phenotypic cellular aspects of innate immunity levels. Methods: This is a Cross-sectional study evaluated 36 women who underwent clinical examination, gynecological examination, transvaginal ultrasound and treatment with laparotomy as indicated pelvic mass. According to the results of these tests, the women selected were grouped into two groups: control, absence of malignancy, with benign ovarian neoplasm and malignant ovarian neoplasm. The serum levels of serum molecules surface expression of cells of the innate immune response with analysis by flow cytometry. Differences between groups were evaluated by the Mann-Whitney (two groups) or Kruskal-Wallis (three groups) as indicated. Differences with p <0.05 were considered significant. Results: We selected 36 patients: 10 women in the control group, 9 in the group of benign ovarian neoplasm and 17 in the group of malignancy. More than 70% of patients with ovarian cancer presented with advanced disease and values of CA125 much changed. For the analysis found, there was a change between the groups for molecules expression in neutrophils... (Complete abstract click electronic access below) / Orientador: Agnaldo Lopes da Silva Filho / Coorientador: Andréa Teixeira de Carvalho / Banca: Paulo Traiman / Banca: Luciana Maria da Silva / Mestre

Resumo: O câncer epitelial do ovário representa um desafio à Oncologia Ginecológica devido ao seu caráter insidioso e alta letalidade. Evidências apontam para o conceito de que o sistema imunológico interage com o tumor em desenvolvimento e pode ser responsável pelo controle do crescimento e regressão tumoral. A resposta imune adaptativa no ambiente tumoral, inclui a imunidade humoral composta de anticorpos produzidos pelas células B e da imunidade celular composta de células T CD4+ e células T CD8+. Este estudo visa avaliar a resposta imune adaptativa sérica em mulheres com neoplasia ovariana. Foram analisadas amostras de sangue periférico obtidas de mulheres hígidas (n=10 - grupo controle), com tumor benigno de ovário (n=9) e com câncer de ovário (n=17). As amostras foram avaliadas pela técnica de citometria de fluxo, onde utilizou-se 5 parâmetros: tamanho celular , complexidade interna e três fluorescências: FITC, PE e TC. O painel de anticorpos monoclonais incluiu os marcadores: CD4, CD8, HLA-DR, CD54, CD62L, CD18, CCR2, CXCR4, CCR5, CCR3, CXCR3, CD25, CD5, CD69, CD19, CD23, e controle isotípico. As diferenças entre os grupos foram avaliadas pelo teste de Mann-Whitney ou Kruskal-Walis conforme indicados. As diferenças com valor de p<0,05 foram consideradas significativas. Houve uma diminuição estatisticamente significativa (p<0,05) da porcentagem de células T do grupo de mulheres com câncer de ovário quando comparado ao grupo controle. A análise dos resultados mostrou que o percentual de linfócitos T CD4+ apresentou diferenças significativas entre os grupos (p=0,0399). Entretanto a população de linfócitos T CD8+ não apresentou diferenças significativas (p=0,2939). A análise de percentual de linfócitos B (CD19+) identificou diferença significativa na comparação entre os três grupos avaliados (p=0,0463). Foi observado uma diminuição do percentual... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: The epithelial ovarian cancer represents a challenge to Gynecologic Oncology due to its insidious nature and high fatality. Evidences show that the immune system interacts with the tumor development and may be responsible for growth control and tumor regression. The adaptive immune response in the tumor environment, includes antibodies produced by B cells and cellular immunity consisting of CD4 + and CD8 + T cells. This study aims to evaluate the adaptive immune response in peripheral blood of women with ovarian cancer. Methods:. We analyzed peripheral blood samples obtained from healthy women (n = 10 - control group) with benign ovarian tumor (n = 9) and ovarian cancer (n = 17). The samples were evaluated by the technique of flow cytometry, where we used 5 parameters: cell size, internal complexity and three fluorescence: FITC, PE and TC. The panel of monoclonal antibodies included markers: CD4, CD8, HLA-DR, CD54, CD62L, CD18, CCR2, CXCR4, CCR5, CCR3, CXCR3, CD25, CD5, CD69, CD19, CD23, and isotype control. Differences between groups were evaluated by the Mann-Whitney or Kruskal- Wallis tests. Differences with p <0.05 were considered significant.Results: There was a significant decrease (p <0.05) of the percentage of T cells in the group of women with ovarian cancer when compared to the control group. The results showed that the percentage of CD4 + T cells showed significant differences between the groups (p = 0.0399). However the population of CD8+ T cells did not show significant differences (p = 0.2939). The analysis of the percentage of B lymphocytes (CD19+) identified a significant difference between the three study groups (p = 0.0463). We observed a decrease in the percentage of B cells of groups of women with benign tumor and ovarian cancer in the control group. Among the adhesion molecules tested... (Complete abstract click electronic access below) / Orientador: Agnaldo Lopes da Silva Filho / Coorientador: Andréa Teixeira de Carvalho / Banca: Paulo Traiman / Banca: Luciana Maria Silva / Mestre

Made available in DSpace on 2014-06-11T19:29:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-26Bitstream added on 2014-06-13T18:59:45Z : No. of bitstreams: 1 paula_soc_me_botfm.pdf: 316334 bytes, checksum: 6b85b467e2fa0ecca5eec101c94687cd (MD5) / O câncer de ovário apresenta diagnóstico tardio e alta letalidade, devido à falta de biomarcadores sensíveis e específicos e à rápida progressão desse câncer, assintomático em estadios iniciais. As células imunes têm a capacidade de eliminar as células malignas e regular a progressão tumoral. Contudo, as células imunes do microambiente tumoral são disfuncionais e falham no controle da expansão tumoral podendo, inclusive, promover o crescimento da neoplasia. Apesar das inúmeras tentativas de se correlacionar o grau e o tipo de infiltrado celular com o prognóstico ou sobrevida do paciente com câncer de ovário, não há consenso sobre o real significado do infiltrado leucocitário nesses casos. Desse modo, este estudo tem como objetivo a ampliação dos conhecimentos relativos à imunidade inata em mulheres com câncer de ovário, através caracterização dos aspectos fenotípicos celulares da imunidade inata sérica. Trata-se de estudo transversal onde foram avaliadas 36 mulheres submetidas a exame clínico, ginecológico, ultrassonografia transvaginal e tratamento com laparotomia nos casos indicados de massa pélvica. De acordo com o resultado desses exames, as mulheres selecionadas foram agrupadas nos grupos: controle- ausência de neoplasia, com neoplasia ovariana benigna e neoplasia ovariana maligna. Realizou-se dosagem sérica de moléculas de expressão de superfície de células da resposta imune inata com análise através da citometria de fluxo. As diferenças entre os grupos foram avaliadas pelo teste de Mann-Whitney (dois grupos) ou Kruskal-Walis (três grupos) conforme indicados. As diferenças com valor de p<0,05 foram consideradas significativas. Foram selecionadas 36 pacientes: 10 mulheres no grupo controle, 9 no grupo de neoplasia ovariana benigna e 17 no grupo de neoplasia maligna. Mais de 70% das pacientes com câncer de ovário apresentavam-se com doença... / Introduction: Ovarian cancer presents late diagnosis and high mortality due to lack of sensitive and specific biomarkers and the rapid progression of this cancer, asymptomatic in early stages. Immune cells have the ability to eliminate malignant cells and regulate tumor progression. However, the immune cells of the tumor microenvironment are dysfunctional and fail to control the tumor growth and may even promote the growth of cancer. Despite numerous attempts to correlate the degree and type of cellular infiltrate and the prognosis or survival of patients with ovarian cancer, there is no consensus about the real meaning of the leukocyte infiltrate in these cases. Thus, this study aims to increase knowledge about the innate immunity in women with ovarian cancer through characterization of phenotypic cellular aspects of innate immunity levels. Methods: This is a Cross-sectional study evaluated 36 women who underwent clinical examination, gynecological examination, transvaginal ultrasound and treatment with laparotomy as indicated pelvic mass. According to the results of these tests, the women selected were grouped into two groups: control, absence of malignancy, with benign ovarian neoplasm and malignant ovarian neoplasm. The serum levels of serum molecules surface expression of cells of the innate immune response with analysis by flow cytometry. Differences between groups were evaluated by the Mann-Whitney (two groups) or Kruskal-Wallis (three groups) as indicated. Differences with p <0.05 were considered significant. Results: We selected 36 patients: 10 women in the control group, 9 in the group of benign ovarian neoplasm and 17 in the group of malignancy. More than 70% of patients with ovarian cancer presented with advanced disease and values of CA125 much changed. For the analysis found, there was a change between the groups for molecules expression in neutrophils... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:29:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-26Bitstream added on 2014-06-13T20:39:42Z : No. of bitstreams: 1 freitas_gf_me_botfm.pdf: 572966 bytes, checksum: 78f32ce23035c30197d379a64992ac2f (MD5) / O câncer epitelial do ovário representa um desafio à Oncologia Ginecológica devido ao seu caráter insidioso e alta letalidade. Evidências apontam para o conceito de que o sistema imunológico interage com o tumor em desenvolvimento e pode ser responsável pelo controle do crescimento e regressão tumoral. A resposta imune adaptativa no ambiente tumoral, inclui a imunidade humoral composta de anticorpos produzidos pelas células B e da imunidade celular composta de células T CD4+ e células T CD8+. Este estudo visa avaliar a resposta imune adaptativa sérica em mulheres com neoplasia ovariana. Foram analisadas amostras de sangue periférico obtidas de mulheres hígidas (n=10 – grupo controle), com tumor benigno de ovário (n=9) e com câncer de ovário (n=17). As amostras foram avaliadas pela técnica de citometria de fluxo, onde utilizou-se 5 parâmetros: tamanho celular , complexidade interna e três fluorescências: FITC, PE e TC. O painel de anticorpos monoclonais incluiu os marcadores: CD4, CD8, HLA-DR, CD54, CD62L, CD18, CCR2, CXCR4, CCR5, CCR3, CXCR3, CD25, CD5, CD69, CD19, CD23, e controle isotípico. As diferenças entre os grupos foram avaliadas pelo teste de Mann-Whitney ou Kruskal-Walis conforme indicados. As diferenças com valor de p<0,05 foram consideradas significativas. Houve uma diminuição estatisticamente significativa (p<0,05) da porcentagem de células T do grupo de mulheres com câncer de ovário quando comparado ao grupo controle. A análise dos resultados mostrou que o percentual de linfócitos T CD4+ apresentou diferenças significativas entre os grupos (p=0,0399). Entretanto a população de linfócitos T CD8+ não apresentou diferenças significativas (p=0,2939). A análise de percentual de linfócitos B (CD19+) identificou diferença significativa na comparação entre os três grupos avaliados (p=0,0463). Foi observado uma diminuição do percentual... / Introduction: The epithelial ovarian cancer represents a challenge to Gynecologic Oncology due to its insidious nature and high fatality. Evidences show that the immune system interacts with the tumor development and may be responsible for growth control and tumor regression. The adaptive immune response in the tumor environment, includes antibodies produced by B cells and cellular immunity consisting of CD4 + and CD8 + T cells. This study aims to evaluate the adaptive immune response in peripheral blood of women with ovarian cancer. Methods:. We analyzed peripheral blood samples obtained from healthy women (n = 10 - control group) with benign ovarian tumor (n = 9) and ovarian cancer (n = 17). The samples were evaluated by the technique of flow cytometry, where we used 5 parameters: cell size, internal complexity and three fluorescence: FITC, PE and TC. The panel of monoclonal antibodies included markers: CD4, CD8, HLA-DR, CD54, CD62L, CD18, CCR2, CXCR4, CCR5, CCR3, CXCR3, CD25, CD5, CD69, CD19, CD23, and isotype control. Differences between groups were evaluated by the Mann-Whitney or Kruskal- Wallis tests. Differences with p <0.05 were considered significant.Results: There was a significant decrease (p <0.05) of the percentage of T cells in the group of women with ovarian cancer when compared to the control group. The results showed that the percentage of CD4 + T cells showed significant differences between the groups (p = 0.0399). However the population of CD8+ T cells did not show significant differences (p = 0.2939). The analysis of the percentage of B lymphocytes (CD19+) identified a significant difference between the three study groups (p = 0.0463). We observed a decrease in the percentage of B cells of groups of women with benign tumor and ovarian cancer in the control group. Among the adhesion molecules tested... (Complete abstract click electronic access below)

Orientador: André Almeida Schenka / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-17T05:00:04Z (GMT). No. of bitstreams: 1 Nicolosi_JacquelineSpacagna_M.pdf: 1558921 bytes, checksum: 35ee58bf52c7629b47f27930a374f9fa (MD5) Previous issue date: 2010 / Resumo: Em neoplasias malignas, a angiogênese parece favorecer não só o crescimento celular como a disseminação sistêmica, tendo, potencialmente, valor diagnóstico, prognóstico e para o desenvolvimento de novas estratégias terapêuticas. Contudo, que este papel é variável de um tumor para outro, o que provavelmente reflete diferenças biológicas entre as neoplasias e limitações ou variações metodológicas dos estudos realizados. Nas neoplasias ovarianas, devido à escassez de trabalhos e grande variabilidade metodológica destes, o papel da angiogênese ainda não está estabelecido. Objetivos: (1) avaliar a influência de diferentes variáveis metodológicas na quantificação da angiogênese em tumores ovarianos; (2) estabelecer funções matemáticas para a conversão de resultados obtidos com variantes metodológicas. Metodologia: em tumores ovarianos diagnosticados no CAISM-Unicamp entre 1997 a 2003, a vascularização identificada por meio imunocoloração foi quantificada através da determinação de densidade microvascular e área endotelial. Foram avaliadas comparativamente as variantes metodológicas: formato de imagem (TIFF vs. JPEG) e número de campos de análise (1, 3, 5, 10, 15 e 20 campos de médio aumento) Resultados e conclusões: O impacto do formato de imagem sobre a quantificação vascular em tumores epiteliais ovarianos pode ser considerada pequena, já que a concordância entre imagens TIFF e JPEG é elevada (ICC>0,75). Constitui exceção o contraste TIFF vs. JPEGs de baixa qualidade, na variável densidade microvascular automática. Nesse caso, a interconversão de valores pode ser realizada através de funções de correção. Finalmente, no universo amostral avaliado, não foi possível caracterizar uma curva com platô de estabilização que permitisse a estimativa de N mínimo de campos capaz de garantir uma análise fidedigna de angiogênese em tumores ovarianos epiteliais / Abstract: In malignant neoplasms, angiogenesis seems to facilitate tumour growth and metastasis, thus, bearing a potential role in diagnostic/prognostic assessment, as well as in the development of new therapeutic strategies. However, the relative importance of such a role is variable from one entity to another, probably reflecting biological differences between tumours, as well as limitations or methodological variations among studies. In ovarian neoplasms, due to paucity of studies and great methodological variability among them, the role of angiogenesis is yet to be established. The present study aims: (1) to evaluate the influence of different methodological variables on angiogenesis quantitation in ovarian tumours and (2) to establish, by linear regression, functions that may be used to interconvert values obtained from different methodological variants. Methods: using ovarian tumours diagnosed in our Academic Women's Hospital (CAISM-Unicamp) from 1997-2003, tumour-related vascularity was detected by CD34 immunostaining and quantified by assessment of microvessel density and endothelial. Furthermore, the following methodological variants were tested: image file format (TIFF vs. JPEG), and total number of analyzed fields (1, 3, 5, 10 and 20 medium-power fields). Results and conclusions: the impact of image format over vascular quantitation was considered small, since the agreement between TIFF and JPEG was found to be high (ICC>0,75). The only exception was represented by the contrast TIFF vs. low quality JPEGs in automatic microvessel density assessments. Even in these cases, interconversion could be achieved using prediction models developed in this study. As for the second objective of the study, given the sampling universe used, we could not properly characterize a stabilization plateau curve which could allow for extrapolation of the minimum number of analysis fields (as required for an accurate morphometric analysis of angiogenesis in ovarian epithelial tumors) / Mestrado / Ciencias Biomedicas / Mestre em Ciências Médicas

Introdução: O câncer de ovário é considerado a primeira causa de óbito entre os tumores ginecológicos. Entretanto, com os avanços nos tratamentos as pacientes com câncer de ovário apresentam uma sobrevida maior, sendo fundamental discutir sua qualidade de vida. Objetivos: Validar e analisar a confiabilidade de dois questionários existentes na literatura, a saber: EORTC-OV28 e FACT-O e avaliar a compreensão, preferência e aceitação dos mesmos. Metodologia: O estudo foi realizado nos Ambulatórios de Ginecologia Oncológica e Oncologia Clínica do Hospital do Câncer AC Camargo São Paulo. Foram analisadas 114 mulheres com diagnóstico de câncer de ovário, em qualquer estádio da doença. Para cada questionário foi analisada a consistência interna (alpha de Cronbach), a validade de constructo convergente (coeficiente de correlação de Spearman com os domínios do questionário SF-36 e HADS), a validade de constructo discriminante (comparação das médias dos escores, segundo estadiamento inicial (I e II) x avançado (III e IV), Karnosfky (80-100 por cento x 5070 por cento ), doença atual (sem evidência de doença X com evidência de doença) e tratamento atual (em tratamento e pós tratamento)) e analisado o grau de compreensão dos mesmos. A reprodutibilidade foi verificada após duas semanas e foi analisada por meio da comparação de médias (Wilcoxon), coeficiente de correlação intra-classe e gráficos de Bland-Altman. Resultados: As escalas de sintomas dos questionários EORTC-OV28 e bem estar familiar do FACT-O, respectivamente, apresentaram valor de alpha de Cronbach 0,85 e 0,79. A maioria das escalas apresentou correlação significativa com os domínios do SF-36 e HADS e foi capaz de discriminar entre os grupos de comparação. Ambos apresentaram boa compreensão. Quanto ao tempo de preenchimento o questionário EORTC-OV 28 apresentou menor média (5,10 min.). Todos os questionários apresentaram bons índices de reprodutibilidade. Conclusões: O questionário EORTC OV 28 apresentou melhores parâmetros de validade, mas as demais análises foram muito semelhantes entre os questionários. A escolha do questionário a ser aplicado pelo pesquisador dependerá dos aspectos considerados por ele relevantes na pesquisa / Introduction: Ovarian cancer is considered the main cause of death among gynecological tumors. Therefore, ovarian cancer patients have presented higher survival rates due to advances in treatments and it becomes necessary to discuss quality of life. Aim: Validate and analyze the reliability of two questionnaires in the literature, namely OV28-EORTC and FACT-O and assess comprehension, preference and acceptance. Methodology: This study was conducted in outpatient clinics of Gynecology Oncology and Medical Oncology at the Cancer Hospital AC Camargo - São Paulo. The total amount of subjects analyzed was 114 women diagnosed with ovarian cancer at any stage of the disease. Moreover, in each questionnaire, internal consistency (Cronbach\'s alpha), the convergent construct validity (Spearman correlation coefficient with domains of the SF-36 and HADS), the discriminant construct validity (comparison of mean scores of the second stage initial (I and II) x advanced (III and IV), Karnosfky (80-100 per cent vs. 50-70 per cent ), current disease (without evidence of disease X with evidence of disease), current treatment (treatment and post treatment) and degree of understanding were analysed. Reproducibility was checked after 2 weeks and analyzed by comparing means (Wilcoxon), coefficient of intraclass correlation and Bland-Altman. Results: The prognostic scales of the EORTC questionnaires OV28-and family welfare of the FACTO, had a Cronbach alpha value of 0.85 and 0.79, respectively. The majority of scales correlated significantly with the SF-36 and HADS; nonetheless, the ability of differentiation between comparison groups could be realised; therefore. both have presented good understanding. The questionnaire EORTC OV-28 had a lower average (5.10 min) in relation to the time spending to complete it. Furthermore, all questionnaires showed good levels of reproducibility. Conclusions: To sum up, the questionnaire EORTC OV 28 presented the best validity parameters, although the analysis were very similar between the questionnaires. Therefore, the investigator should firstly, consider which are the required aspects to evaluate to be able to make the correct choice

Philosophiae Doctor - PhD / Ovarian cancer is called the “Silent Killer” as it is often diagnosed in advanced stages of the disease or misdiagnosed which ends with a poor prognostic outcome for the patient. A high rate of disease relapse, a high incidence-to-mortality ratio as well as acquired multidrug resistance makes it necessary to find alternative diagnostic- and therapeutic tools for ovarian cancer. Nanotechnology describes molecular devices with at least one dimension in the sub- 1μm scale and has been suggested as a possible solution for overcoming challenges in cancer multidrug resistance as well as early diagnosis of the disease. One-pot synthesized gold nanoparticles were used to demonstrate in vitro drug delivery of doxorubicin in a manner which overcame the cytoprotective mechanisms of a multidrug resistant ovarian carcinoma cell line (A2780cis) by inducing apoptosis mediated by caspase-3 within 3h of treatment. The gold nanoparticles were further functionalized with nitrilotriacetic acid and displayed specific interaction with a 6xHis-tagged cancer targeting peptide, chlorotoxin. Proprietary indium based quantum dots were functionalized with the same surface chemistry used for gold nanoparticles and bioconjugated with chlorotoxin. Wide field fluorescence studies showed the peptide-quantum dot construct specifically targeted enhanced green fluorescent tagged matrix metalloproteinase-2 transfected A2780cis cells in a specific manner. The cytoprotective multidrug resistant mechanisms of the ovarian carcinoma was overcome successfully with a single dose of doxorubicin loaded gold nanoparticles and tumour specific targeting was demonstrated using quantum dots with a similar surface chemistry used for the gold nanoparticles.

Ovarian cancer (OC) accounts for 4% of all cancer cases and 4.2% of all cancer deaths worldwide. OC is the most lethal gynecological cancer because it lacks early disease symptoms and does not have a specific diagnostic marker. As a result, more than 70% of OC patients are diagnosed in later stages when the disease has already metastasized and the 5-year survival rate has decreased to less than 20% compared with approximately 90% survival for women diagnosed with early stage disease. Therefore, I initiated my studies with a computational analysis of the 27 most commonly reported literature-derived ovarian cancer (LDOC) protein biomarkers. I found that LDOC protein biomarkers share many biochemical features including a preponderance for a stable protein structure, the ability to be secreted, and functionality related to extracellular matrix (ECM) modification, immune response and/or energy production. Subsequently, I analyzed the human proteome to identify proteins that also share these biochemical features. Of the 70,616 proteins in the human proteome, 683 proteins were found to have similar biochemical features to the 27 LDOC proteins. I also identified a subset of 21 potential additional protein regulators of ovarian cancer (APROC) that interact with LDOCs. Three of the APROCs identified were amylase proteins AMY1A, AMY2A, and AMY2B which cleaves alpha 1, 4-glycosidic bonds in polysaccharides. Amylase is reportedly overexpressed in and secreted by ovarian tumors but its functional contribution to OC remains unknown[1]. In this thesis, I posit that amylase contributes to OC invasion. I initiated my studies by computational characterizing the different amylase isozymes to predict which amylase isozyme(s) is most likely overexpressed in and contributory to OC invasion. I found that AMY1 and AMY2B have unique regions of disorder and unique phosphorylation sites indicating that AMY1 and AMY2B would be more likely to interact with other proteins, and to be easily secreted. Using OC patient serum samples, I was able to validate AMY1 and AMY2B overexpression by western immunoblotting. I then developed an in vitro model system to study the molecular contribution of amylase to OC invasion using normal ovarian surface epithelial (IOSE) and OC cell lines. I showed that OC cells generally overexpress and secrete metabolically active amylase isozymes AMY1 and AMY2B. Abrogating amylase activity using siRNA silencing technology decreased the capacity of OC cells to invade collagen coated Boyden chambers and increased sulfated glycosaminoglycans (sGAG) production. Since a survey of OC cell lines indicated that cancer cells have a bulkier glycocalyx compared to IOSE cells and immunogold labeling studies indicated the presence of amylase within the immediate OC microenvironment, my data suggest that, by cleaving alpha 1, 4-glycosidic bonds in glycoconjugates present within ECM, amylase may remodel the ECM to promote an invasive cancer phenotype. Amylase is therefore a target for therapeutic intervention in OC patients with hyperamylasemia. I established Spirulina, a dietary supplement, as a novel transcriptional inhibitor of amylase. Spirulina inhibited amylase expression in OC cell lines at both the message and protein levels. Spirulina reduced OC cell invasion and migration in vitro, putatively by decreasing amylase expression.

Class of 2017 Abstract / Objectives: The specific aims of this project were to evaluate the incidence of carboplatin HSR in patients with the BRCA1 or BRCA2 mutations compared to those without these mutations. Secondary objectives were to identify carboplatin cycles where reactions occurred, grade of reaction, and treatment outcomes. Methods: This retrospective chart review included 167 ovarian, fallopian tube, and primary peritoneal cancer patients at the University of Arizona Cancer Center who underwent a regimen with carboplatin from 2013-2015. Results: 126 out of 167 patients were analyzed. HSR occurred in 4 patients with BRCA mutations, and in 9 patients without mutations, though incidence was not significant with respect to the groups (3.1% versus 17.4%, P=0.5291). Overall, there were 11 grade 1 reactions, 14 grade 2 reactions, and 16 grade 3 reactions to carboplatin. Conclusions: Presence of a BRCA1/2 mutation was not associated with a higher incidence of HSR in carboplatin. More studies are needed to clarify the impact of BRCA mutations on developing carboplatin HSR.