O câncer de próstata (CaP) é o tumor maligno mais frequente e uma das principais causas de morte por câncer na população masculina no mundo e no Brasil. Três fatores são de fundamental importância no prognóstico da doença: o estadiamento, o grau histológico (avaliado pelo escore de Gleason) e o antígeno prostático específico (Prostate Specific Antigen - PSA). Há fatores adicionais descritos que podem influenciar na evolução do câncer, como invasão perineural, invasão angiolinfática, acometimento da cápsula, lateralidade do tumor, estágio tumoral e invasão de linfonodos. Embora a associação entre mutações do BRCA2 e o risco de câncer de próstata esteja bem documentado, pouco se sabe sobre o papel do BRCA2 na progressão do câncer de próstata após o diagnóstico inicial. O presente trabalho se propõe a melhor elucidar o papel da proteína BRCA2 nos carcinomas prostáticos comparando os resultados da expressão proteica de BRCA2 com dados clinicopatológicos de pacientes acometidos por Câncer de Próstata, além de avaliar a expressão de BRCA2 com escore de Gleason >=7 após a prostatectomia, tanto na classificação de Gleason segundo Epstein 2005, quanto na classificação do grau de Gleason definida em 2014 com suas respectivas biópsias por agulha. Dos arquivos do Serviço de Patologia do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo, foram selecionados 125 blocos de parafina com amostras de câncer de próstata diagnosticados entre 2005 e 2010. As lâminas coradas com hematoxilina e eosina (H&E) foram utilizadas para a construção de microarranjos teciduais (TMAs). Nesses TMAs foi realizado estudo imunohistoquímico para BRCA2. A proteína BRCA2 com ponto de corte de 15% (classificação antiga) foi superexpressa em 118 casos (94,4%), enquanto a expressão de BRCA2 com cut-off de 56% (classificação nova) foi expressa em 63 (51,2%) de 125 casos. A superexpressão de BRCA2 associada a classificação antiga de Gleason correlacionou-se com bilateralidade do tumor e com estágio tumoral, em contrapartida a expressão de BRCA2 associada a atual classificação se correlacionou apenas com lesão intraepitelial prostática (PIN). Nossos resultados indicam que a expressão de BRCA2 pode ser um biomarcador importante de progressão tumoral nos carcinomas de próstata. / Prostate cancer (PCa) is the most common malignancy and a major cause of cancer death in male population in the world and in Brazil. Three factors are of fundamental importance in the prognosis of the disease: the staging, histological grade (measured by Gleason score) and the Prostate Specific Antigen (PSA). There are additional factors described that may influence the development of cancer, such as perineural invasion, angiolymphatic invasion, capsule involvement, tumor laterality, tumor stage and lymph node invasion. Although the association between mutations in Breast Cancer Gene 2 (BRCA2) and the risk of prostate cancer is well documented, little is known about the role of BRCA2 in the progression of prostate cancer after initial diagnosis. This study aims to elucidate the role of BRCA2 protein in prostate carcinomas comparing the results of the BRCA2 protein expression with clinicopathological data of patients with prostate cancer. For that reason, we evaluated the expression of BRCA2 with Gleason score >=7 after prostatectomy, both in the second Epstein 2005 Gleason score, and in the classification of Gleason grade set in 2014 with their respective needle biopsies. 125 paraffin blocks with prostate cancer samples diagnosed between 2005 and 2010 were selected from the archives of Hospital Pathology Service of the Ribeirao Preto Medical School, University of Sao Paulo. The slides stained with hematoxylin and eosin (H&E) were used for the construction of the tissue microarrays (TMAs). These TMAs was performed immunohistochemical study for BRCA2. The BRCA2 protein with a cut-off point of 15% (old classification) was overexpressed in 118 cases (94.4%), while the expression of BRCA2 to cut off 56% (new classification) was expressed in 63 (51.2%) of 125 cases. The BRCA2 associated overexpression of the old classification of Gleason presented correlation with tumor bilateralism and tumor stage, however BRCA2 expression associated with the current classification was correlated only with Prostatic Intraepithelial Lesion (PIN). Our results indicate that BRCA2 expression can be an important marker of tumor progression in prostate carcinomas.

Dentre as diversas formas de se tratar o câncer de próstata, a braquiterapia com sementes de iodo-125 é uma opção que apresenta ótimos resultados e menor ocorrência de efeito colateral. No presente trabalho diferentes métodos de deposição de iodo radioativo em substrato de prata foram comparados com o propósito de eleger a alternativa mais adequada para a produção rotineira de sementes de iodo-125 do IPEN. A metodologia utilizada foi escolhida com base na infraestrutura disponível e na experiência dos pesquisadores presentes. Por essa razão, utilizou-se o iodo-131 para realização dos testes (mesmo comportamento do iodo-125). Quatro métodos foram selecionados: Método 1 (teste de eletrodeposição baseado no método desenvolvido por D. Kubiatowicz) com a eficiência de 65,16%; Método 2 (Reação química baseada no método desenvolvido por D. Kubiatowicz - HCl) com o resultado de 70,80% de eficiência; Método 3 (Reação química baseada no método desenvolvido pela Dra Maria Elisa Rostelato aquecimento/sulfeto) com 55,80% de eficiência; Método 4 (IQ-IPEN) apresentou o melhor resultado de eficiência, 99%. Como há mais fixação do material radioativo (que representa praticamente todo o custo da semente) por esse método, o preço final é o mais barato, sendo esse o método sugerido para ser implementado no laboratório de produção de fontes de braquiterapia do IPEN. Além disso o método é o mais rápido. / Among the different ways to treat prostate cancer, brachytherapy with iodine- 125 seeds is an option that provides good results and fewer side effects. In the present study several deposition methods of radioactive iodine in a silver substrate were compared in order to choose the most suitable alternative for the routine production to be implemented at IPENs laboratory. The methodology used was chosen based on the available infrastructure and experience of the researchers present. Therefore, the I131 was used for testing (same chemical behavior as I131). Four methods were selected: Method 1 (test based on electrodeposition method developed by D.Kubiatowicz) presented 65.16% efficiency; Method 2 (chemical reaction based on the method developed by D. Kubiatowicz - HCl) with the result of 70.80% efficiency; method 3 (chemical reaction based on the method developed by Dr. Maria Elisa Rostelato) with 55.80% efficiency; Method 4 (IQ-IPEN) resulted in 99% efficiency. Since this method has more radioactive material fixation (which represents virtually the entire cost of the seed), the final price is the cheapest. This method is the suggested one to be implemented in the IPENs laboratory for brachytherapy sources production. Besides, the method is the fasted one.

A tomografia por emissão de pósitrons associada à tomografia computadorizada (PET/CT) é um dispositivo que combina as características de medicina nuclear (PET) e de radiologia (CT) obtendo imagens metabólicas (PET) e anatômicas sobrepostas (CT). Combinando as duas tecnologias de exames, o exame PET / CT permite aos médicos diagnosticar com maior precisão e identificar o câncer, doenças cardíacas e distúrbios cerebrais. O radiofármaco 18FFAc (fluoroacetato) é promissor para a detecção de tumores primários de próstata e de mama, utilizando a técnica de PET/CT. Estudos recentes mostram a eficácia do 18F-FAc na detecção de tumores que têm baixa captação de 18F-FDG (fluordesoxiglicose). O fluoroacetato é um substrato para a acetil-CoA sintase, enzima que metaboliza ácido fluorocitrato que não é mais metabolizado, levando à inibição da aconitase e do ácido tricarboxílico. O objetivo deste trabalho foi desenvolver um radiofármaco emissor de pósitron, 18F-FAc no IPEN-CNEN/SP em um acordo com o Hospital AC-Camargo / São Paulo. O íon fluoreto (18F-) foi produzido, usando os cíclotrons Cyclone 30 e 18 da IBA localizados no IPEN-CNEN/SP, através da irradiação de água enriquecida em 18O com prótons e dose integrada de 30μAh. A marcação do 18F-FAc foi realizada em um módulo de síntese TRACERlab MXFDG (GE), utilizando kits adquiridos da ABX. O controle de qualidade radioquímico de 18F-FAc foi realizado por cromatografia em camada fina TLC-SG 25 folhas de alumínio em tiras (1,5 x 12 cm ) usando como solvente clorofórmio:metanol (1:1). Para o controle de qualidade radionuclídico, amostras de 18F-FAc e 18F-Fluoreto foram analisadas por espectroscopia de raios-gama. A avaliação dos solventes residuais foi realizada por cromatografia em fase gasosa e a análise de kryptofix foi realizada por TLC utilizando tiras de TLC-SG, metanol:clorofórmio (9:1) como solvente e padrões de kryptofix 2.2.2. Os estudos de biodistribuição foram realizados com 18FFAc injetado em camundongos swiss sadios. Um procedimento reprodutível foi desenvolvido para o preparo do 18F-FAc com um rendimento de marcação de 37% (não corrigido) e 52% (corrigido para o decaimento) e estabilidade de 19 horas. A análise de controle de qualidade mostrou que o produto tinha as exigências adequadas para utilização, com pureza radioquímica superior a 99,9%. Os estudos de biodistribuição em animais sadios mostraram a esperada captação em todos os órgãos medidos com eliminação renal e intestinal. / PET / CT (positron emission tomography / computed tomography) is a device that combines the features of diagnostic nuclear medicine (PET) and Radiology (CT) superimposing metabolic (PET) and anatomical (CT) images. By combining the two technologies examinations, the PET/CT scan allows physicians to diagnose more accurately and identify cancer, heart disease and brain disorders. The radiopharmaceutical 18F-FAc (fluoroacetate) is promising for application in detection of primary tumors of prostate and breast, using PET-CT techniques. Recent studies are showing the efficacy of the 18F-FAc in the detection of tumors that have low uptake of 18F-FDG (fluorodeoxyglucose). The fluoroacetate is a substrate for the enzyme acetyl-CoA synthase that metabolizes acid fluorcitric that, not being metabolized, causes inhibition of aconitase and inhibition of tricarboxylic acid. The aim of this work was to develop a positron emitting radiopharmaceutical, 18F-FAc at IPEN-CNEN/SP in agreement with Hospital AC-Camargo/ São Paulo. The 18F-fluoride ion was produced using the Cyclone 30 and 18 cyclotrons from IBA located at IPEN-CNEN/SP, by irradiating enriched 18O water with protons with integrated dose 30μAh. The labelling of 18F-FAc was performed in a synthesis module TRACERlab MXFDG (GE), using kits purchased from ABX. The radiochemical quality control of 18F-FAc was performed by Thin Layer Chromatography using TLC-SG 25 aluminium sheets strips (1.5 x 12 cm) and chloroform:methanol (1:1) as the solvent. For the radionuclidic quality control, samples of 18F-FAc and 18F-Fluoride were analysed by gama-ray spectroscopy. The evaluation of the residual solvents was performed by gas chromatography and the analysis of kryptofix was performed by TLC using TLC-SG strips, methanol:chloroform (9:1) as solvent and kryptofix 2.2.2 standards. Biodistribution studies were performed with 18F-FAc injected into healthy Swiss mice. A reliable procedure was developed for preparation of 18F-FAc with a labelling yield of 37% (uncorrected) and 52% (corrected for decay) and stability of 19 hours. The quality control analysis showed that the product had the proper requirements for use, with radiochemical purity exceeding 99.9%. The biodistribution studies in healthy animals showed the expected uptake results in all the measured organs with intestinal and renal elimination.

Background and aims of the study. Prostate cancer (PCa) is one of the most common hormone-dependent cancers in men in Western and also Asian countries. The standard treatment options for localized PCa include surgery and androgen-deprivation therapy (ADT). However, most patients upon ADT therapy invariably relapse and progress to a more aggressive and metastatic stage termed as castration-resistant PCa (CRPC). Accumulating studies indicate that androgen receptor (AR) transcriptional activity is dysregulated during the advanced progression of CRPC. One important mechanism responsible for the growth of CRPC includes increased intra-tumoral androgen synthesis in PCa. Recently, a novel androgen-responsive fusion gene TMPRSS2:ERG formed by fusion between the transmembrane protein TMPRSS2 and transcription factor ERG, has been identified in approximately 50% PCa samples, which results in the aberrant expression of ERG function as oncogenic factor in PCa. Currently, TMPRSS2:ERG is regarded as a significant potential diagnostic and prognostic biomarker for PCa. Estrogen-related receptor alpha-ERRα, the first identified ligand-independent orphan nuclear receptor, is characterized to be up-regulated in advanced cancers, suggesting that ERRα might play important regulatory roles in the malignant progression of PCa. Previous studies showed that ERRα can functionally cross-talk with AR signaling via co-targeting to AR targets and regulate the expression of some steroidogenic enzymes in breast cancer. Based on this background, it is hypothesized that ERRα could functionally regulate the TMPRSS2:ERG fusion gene and play a regulatory role in the development and progression of CRPC through activation of the intracellular androgen synthesis pathway. / Results. 1) The results obtained in this study showed that suppression of ERRα by its specific inverse agonist XCT790 or shRNA-knockdown could induce down-regulation of TMPRSS2:ERG and also its target genes in AR-positive VCaP PCa cells. 2) Ectopic expression of ERRα and/or its coactivator PGC-1α could increase the expression of TMPRSS2:ERG in AR-negative NCI-H660 PCa cells. 3) Two ERRα-DNA binding elements were identified by ChIP assay and sequence analysis in the promoter of TMPRSS2:ERG and both of these two elements could be transactivated by ERRα and PGC-1α. 4) Ectopic expression of TMPRSS2:ERG under the regulation of ERRα enhanced the prostatic cell invasion capacity as shown in the TMPRSS2:ERG infectants of BPH-1 and PC-3 prostatic cells. 5) ERG expressed by the TMPRSS2:ERG fusion could directly transactivate the ERRα gene in prostatic cells. 6) A positive correlation on the expressions between TMPRSS2:ERG and ERRα was demonstrated in a xenograft model of CRPC (VCaP-CRPC). 7) The expression of TMPRSS2:ERG and ERRα showed significant up-regulation and the transactivation activity of ERRα was also enhanced in castration-resistant VCaP-CRPC cells. 8) Ectopic expression of ERRα could promote resistant growth capacity to androgen-deprivation condition in LNCaP PCa cells, whereas shRNA-mediated silence of ERRα could weaken this resistant capacity. Furthermore, ectopic expression of ERRα in LNCaP-ERRα infectants could promote their in vivo growth resistance to castration in SCID mice. 9) Expression of several androgenic enzyme genes, including CYP11A1, CYP17A1 and ARK1C3, were detected to be up-regulated in castration-resistant VCaP-CRPC cells. Moreover, ectopic expression of ERRα could induce the increased expression of these enzyme genes in LNCaP-ERRα infectants, whereas knockdown of ERRα by shRNA could decrease their expression. 10) ERRα could directly transactivate the gene promoters of CYP11A1, CYP17A1 and ARK1C3 which contain ERRE elements prediction by sequence analysis. These results suggested that ERRα could play a role in de novo or intra prostatic androgen synthesis in the PCa cells. / Conclusions. The results obtained in this study suggested that ERRα and TMPRSS2:ERG could form a positive reciprocal loop in PCa cells, and ERRα could also promote the resistant growth capacity of PCa cells resistant to the androgen-deprivation condition in vitro and also castration-resistant growth in vivo via a mechanism of up-regulation of androgenic enzyme genes. The results also suggested that ERRα might play a significant regulatory role in the development and progression of PCa, particularly the advanced CRPC, and also ERRα could be a potential therapeutic target for the treatment of PCa, particularly the advanced PCa-CRPC. / 研究背景與研究目的：前列腺癌作為激素依賴的一種癌症，經常出現在西方和亞洲國家的男性人群中。對於局限性前列腺癌多採用外科手術和去勢的治療。但是大多數病人經過去勢治療后會再次復發並且形成更加惡心幾轉移的前列腺癌，稱之為去勢難治性前列腺癌（CRPC）。越來越多的研究表明在去勢難治性前列腺癌發病過程中，雄激素受體轉錄活性異性增強。其中一個重要機理解釋為前列腺癌細胞自身合成的雄激素增多。進來，在大約50%的前列腺癌病人中新檢測到一個受雄激素受（AR）體調控的融合基因TMPRSS2:ERG，它是由稱為TMPRSS2的一個跨膜蛋白和一個稱為ERG的轉錄因子融合而成，它的出現導致了在前列腺癌中異常的稱為致癌因子的ERG蛋白的高表達。目前，TMPRSS2:ERG已經被作為一個重要的潛在的診斷和預測的標誌物應用在前列腺癌中。作為第一個鑒定的配體不依賴的孤兒受體-ERRα，被證明在晚期的癌症中有很高的表達，預示著ERRα可能在惡性的癌症中起到一個非常重要的調控作用。之前的研究表明通過共同調控AR的下游基因，ERRα同AR信號通路之間有功能性的交叉調控；除此之外，在乳腺癌中，ERRα還可以調控一些類固醇類化合物的合成相關的一些酶的合成。依據上述，我們推定ERRα可能功能性地調控TMPRSS2:ERG融合基因的表達並且通過調控細胞內的雄激素的合成進而在去勢難治性前列腺癌的發生和發展中起到一個非常重要的作用。 / 結果：本論文研究結果總結如下：1）在有AR表達的前列腺癌細胞-VCaP細胞中，通過ERRα特異性的抑制劑XCT790處理或者shRNA介入的干擾ERRα的mRNA的方法來抑制ERRα，下調了TMPRSS2:ERG和它的一些下游調控基因的表達。2）在沒有AR表達的前列腺癌細胞-NCI-H660細胞中，上調ERRα或者它的特異性的共激活因子PGC-1α表達可以提升TMPRSS2:ERG的表達。3）通過ChIP實驗，在TMPRSS2:ERG的啟動子上面，兩個ERRα的DNA結合位點被鑒定出來。並且這兩個位點可以被ERRα和PGC-1α轉錄激活。4）在兩個前列腺細胞BPH-1和PC-3細胞中，在ERRα的調控下高表達TMPRSS2:ERG融合基因可以增強細胞的侵襲能力。5）融合基因TMPRSS2:ERG導致的ERG蛋白的表達可以直接轉錄激活ERRα的表達。6）我們通過VCaP細胞的異種移植建立VCaP-CRPC的體內模型來模擬CRPC過程，在整個過程中，我們發現TMPRSS2:ERG和ERRα有一致性的表達相關性。除此之外，我們根據上述動物模型通建立了VCaP-CRPC細胞系，並且發現在VCaP-CRPC細胞細胞中，TMPRSS2:ERG和ERRα都有被上調並且ERRα的轉錄活性同樣也提升。7）在LNCaP細胞中高表達ERRα可以提升細胞在去除雄激素的環境中生長的能力。但是當在LNCaP細胞中用shRNA干擾掉ERRα可以明顯減弱這種生長的能力。用LNCaP-ERRα穩轉ERRα的細胞異種移植建立SCID老鼠體內腫瘤模型，我們發現和LNCaP-pBABE對照組相比，LNCaP-ERRα細胞生長的更快更大。並且在對老鼠進行睪丸切除術后，LNCaP-ERRα組細胞更快適應這種環境并繼續生長，相比之下，LNCaP-pBABE對照組則持續萎縮減小。8）在上述的VCaP-CRPC細胞中，我們發現一些和雄激素合成相關的關鍵的酶包括CYP11A1，CYP17A1和ARK1C3的表達量有顯著地提升。而且在LNCaP-ERRα細胞中同樣檢測到這些酶的表達量的提升。然而當在LNCaP細胞中用shRNA干擾掉ERRα可以明顯減降低上述酶的表達。9）我們在CYP11A1，CYP17A1和ARK1C3基因的啟動子區域發現有ERRα結合位點，並且發現這些位點可以被ERRα轉錄激活。 / 結論：本論文的研究結果提示在前列腺癌細胞中，ERRα和TMPRSS2:ERG可以形成一個相互正向調控的循環。除此之外，上調ERRα可以促進細胞在去除雄激素的環境中生長的能力，並且在動物體內可以提升細胞在睪丸去除的環境中的適應和生長能力。這種體內和體外的能力的提升是通過一種潛在的上調前列腺癌細胞的雄激素合成相關的關鍵的酶的表達，進而提升雄激素的含量而得以實現的。上述的結果預示著ERRα可能在前列腺癌發生機發展的過程中起到非常重要的調控作用，尤其在晚期的CRPC中。同時，ERRα也可能作為一個潛在的重要的前列腺癌尤其是晚期的CRPC的治療靶點，尤其是一些潛在ERRα的特異性抑制劑，比如XCT790，可能作為將來用以作為治療前列腺癌的特異性靶點藥物。 / Xu, Zhenyu. / Thesis Ph.D. Chinese University of Hong Kong 2014. / Includes bibliographical references (leaves 126-143). / Abstracts also in Chinese. / Title from PDF title page (viewed on 05, October, 2016). / Xu, Zhenyu. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only.

A tomografia por emissão de pósitrons associada à tomografia computadorizada (PET/CT) é um dispositivo que combina as características de medicina nuclear (PET) e de radiologia (CT) obtendo imagens metabólicas (PET) e anatômicas sobrepostas (CT). Combinando as duas tecnologias de exames, o exame PET / CT permite aos médicos diagnosticar com maior precisão e identificar o câncer, doenças cardíacas e distúrbios cerebrais. O radiofármaco 18FFAc (fluoroacetato) é promissor para a detecção de tumores primários de próstata e de mama, utilizando a técnica de PET/CT. Estudos recentes mostram a eficácia do 18F-FAc na detecção de tumores que têm baixa captação de 18F-FDG (fluordesoxiglicose). O fluoroacetato é um substrato para a acetil-CoA sintase, enzima que metaboliza ácido fluorocitrato que não é mais metabolizado, levando à inibição da aconitase e do ácido tricarboxílico. O objetivo deste trabalho foi desenvolver um radiofármaco emissor de pósitron, 18F-FAc no IPEN-CNEN/SP em um acordo com o Hospital AC-Camargo / São Paulo. O íon fluoreto (18F-) foi produzido, usando os cíclotrons Cyclone 30 e 18 da IBA localizados no IPEN-CNEN/SP, através da irradiação de água enriquecida em 18O com prótons e dose integrada de 30μAh. A marcação do 18F-FAc foi realizada em um módulo de síntese TRACERlab MXFDG (GE), utilizando kits adquiridos da ABX. O controle de qualidade radioquímico de 18F-FAc foi realizado por cromatografia em camada fina TLC-SG 25 folhas de alumínio em tiras (1,5 x 12 cm ) usando como solvente clorofórmio:metanol (1:1). Para o controle de qualidade radionuclídico, amostras de 18F-FAc e 18F-Fluoreto foram analisadas por espectroscopia de raios-gama. A avaliação dos solventes residuais foi realizada por cromatografia em fase gasosa e a análise de kryptofix foi realizada por TLC utilizando tiras de TLC-SG, metanol:clorofórmio (9:1) como solvente e padrões de kryptofix 2.2.2. Os estudos de biodistribuição foram realizados com 18FFAc injetado em camundongos swiss sadios. Um procedimento reprodutível foi desenvolvido para o preparo do 18F-FAc com um rendimento de marcação de 37% (não corrigido) e 52% (corrigido para o decaimento) e estabilidade de 19 horas. A análise de controle de qualidade mostrou que o produto tinha as exigências adequadas para utilização, com pureza radioquímica superior a 99,9%. Os estudos de biodistribuição em animais sadios mostraram a esperada captação em todos os órgãos medidos com eliminação renal e intestinal. / PET / CT (positron emission tomography / computed tomography) is a device that combines the features of diagnostic nuclear medicine (PET) and Radiology (CT) superimposing metabolic (PET) and anatomical (CT) images. By combining the two technologies examinations, the PET/CT scan allows physicians to diagnose more accurately and identify cancer, heart disease and brain disorders. The radiopharmaceutical 18F-FAc (fluoroacetate) is promising for application in detection of primary tumors of prostate and breast, using PET-CT techniques. Recent studies are showing the efficacy of the 18F-FAc in the detection of tumors that have low uptake of 18F-FDG (fluorodeoxyglucose). The fluoroacetate is a substrate for the enzyme acetyl-CoA synthase that metabolizes acid fluorcitric that, not being metabolized, causes inhibition of aconitase and inhibition of tricarboxylic acid. The aim of this work was to develop a positron emitting radiopharmaceutical, 18F-FAc at IPEN-CNEN/SP in agreement with Hospital AC-Camargo/ São Paulo. The 18F-fluoride ion was produced using the Cyclone 30 and 18 cyclotrons from IBA located at IPEN-CNEN/SP, by irradiating enriched 18O water with protons with integrated dose 30μAh. The labelling of 18F-FAc was performed in a synthesis module TRACERlab MXFDG (GE), using kits purchased from ABX. The radiochemical quality control of 18F-FAc was performed by Thin Layer Chromatography using TLC-SG 25 aluminium sheets strips (1.5 x 12 cm) and chloroform:methanol (1:1) as the solvent. For the radionuclidic quality control, samples of 18F-FAc and 18F-Fluoride were analysed by gama-ray spectroscopy. The evaluation of the residual solvents was performed by gas chromatography and the analysis of kryptofix was performed by TLC using TLC-SG strips, methanol:chloroform (9:1) as solvent and kryptofix 2.2.2 standards. Biodistribution studies were performed with 18F-FAc injected into healthy Swiss mice. A reliable procedure was developed for preparation of 18F-FAc with a labelling yield of 37% (uncorrected) and 52% (corrected for decay) and stability of 19 hours. The quality control analysis showed that the product had the proper requirements for use, with radiochemical purity exceeding 99.9%. The biodistribution studies in healthy animals showed the expected uptake results in all the measured organs with intestinal and renal elimination.

INTRODUÇÃO: O câncer de próstata (CAP) é a neoplasia mais comum em homens (excluindo câncer de pele não-melanoma) com mais de 190.000 casos novos esperados em 2010 nos Estados Unidos sendo que mais de 27.000 morrerão em desta doença. No Brasil, segundo o Instituto Nacional do Câncer, a estimativa é em torno de 50 mil casos novos de CAP para 2010. OBJETIVOS: Avaliar a experiência do rastreamento para CAP realizado pelo Hospital de Câncer de Barretos através de uma Unidade Móvel de Prevenção de Câncer (UMPC), e verificar qual o impacto deste rastreamento no estádio clínico em comparação com pacientes diagnosticados e/ou encaminhados ao HCB. MATERIAL E MÉTODO: De janeiro de 2004 a dezembro de 2007, realizou-se rastreamento de CAP em voluntários acima de 45 anos através da UMPC em localidades com difícil acesso à saúde. Foram convocados homens com pelo menos um destes três critérios a seguir: a) PSA sérico = 4,0 ng/ml, b) toque retal suspeito, ou c) PSA entre 2,5 e 4,0 ng/ml com relação PSA livre/total (rPSAl/t) = 15%. Para se avaliar o impacto do rastreamento no estádio clínico ao diagnóstico dos pacientes portadores de CAP, analisaram-se os dois grupos. O grupo I inclui casos de CAP diagnosticados de janeiro de 2005 a dezembro de 2007, através da UMPC. O grupo II inclui pacientes com CAP atendidos pelo HCB no mesmo período, que não haviam feito diagnóstico pela UMPC; e foram encaminhados ao HCB por médicos de especialidades diversas, devido a PSA elevado e/ou TR suspeito realizado na xvii localidade de origem ou a diagnóstico histológico confirmado de CAP. A revisão de prontuários para os grupos realizou-se no serviço de arquivo médico (SAME) e utilizou-se a mesma ficha de coleta de dados, priorizando TNM, PSA e escore de Gleason. Os grupos I e II foram comparados com relação a estadiamento (TNM), PSA e escore de Gleason e feita análise estatística destes dados. RESULTADOS: De janeiro de 2004 a dezembro de 2007, foram rastreados 17.571 homens de 231 cidades brasileiras. Destes, 71,4% nunca tinham feito toque retal e 70,9% nunca fizeram PSA anteriormente. Foram biopsiados 1.647, 904 devido a PSA = 4,0 ng/ml (54,9%), 324 devido a TR suspeito (19,7%), 117 devido a alteração simultânea de ambos os anteriores (7,1%) e 302 quando a relação foi = 15% com PSA entre 2,5 e 3,9 ng/ml (18,3%). Foram diagnosticados 652 casos de CAP (3,7%). Destes, 609 (93,4%) clinicamente localizados (T1-2) e 43 (6,6%) foram T3-4. Na avaliação radiológica e cintilográfica, 26 (4%) eram N1 e 18 (2,8%) eram M1. Comparando-se os grupos, observou-se valores de PSA mais baixos (p<0.001), estadiamento clínico mais favorável (p<0,001), e escore de Gleason com menor grau para o grupo I (p<0.001). CONCLUSÕES: A UMPC mostrou ser uma ferramenta importante para se rastrear populações com acesso médico precário em um país com grande extensão territorial e desigualdades socioeconômicas como o Brasil. O rastreamento mostrou melhoria estatisticamente significativa do estadiamento clínico ao diagnóstico em relação aos pacientes diagnosticados na rotina do Hospital de Câncer de Barretos / BACKGROUND: Prostate cancer (PC) is as the most common neoplasm in men (excluding skin cancer non-melanoma) with more than 190,000 new cases expected in 2009 in the United States and more than 27,000 will die from the disease. In Brazil, according to the Brazilian National Cancer Institute, the estimate is almost 50.000 new cases for 2009. OBJECTIVES: To assess the experience of PC screening conducted by the Barretos Cancer Hospital (BCH) through a mobile cancer prevention unit (MCPU), and to verify the impact of screening on clinical stage compared with patients diagnosed and/or referred to the HCB. MATERIAL AND METHODS: From January 2004 to December 2007, a PC screening was applied to volunteers over 45 years old through a MCPU which reached Brazilian locations with difficult access to health. Men with at least one of the following three criteria were called for further evaluation: a) serum PSA level = 4.0 ng/ml, b) suspicious digital rectal examination (DRE), or c) PSA level of 2.5-4.0 ng/mL and a percent-free PSA (%fPSA) level =15%. To assess the impact of screening on clinical stage at diagnosis of patients screened and non-screened, the men were analyzed in two groups. The PC cases screened from January 2005 to December 2007 through the MCPU constituted Group I. Comprising group II, there was patients with PC treated by BCH in the same period, who hadnt been diagnosed by the MCPU. These patients in group II were referred to hospital by xix physicians of several specialties, mainly due to elevated serum PSA and/either suspicion DRE or histological diagnosis of PC. The data for both groups was held in the medical records and used the same form, prioritizing the data for the TNM staging, PSA and Gleason score. Groups I and II were compared with concern to staging (TNM), PSA and Gleason score and the statistical analysis of these data was performed. RESULTS: From January 2004 to December 2007, 17,571 men from 231 Brazilian cities were screened. Among them 71.4% had never been submitted to DRE examination and 70.9% never had a PSA test. 1,647 men were submitted to biopsy, 904 due to PSA = 4.0 ng/ml (54.9%), 324 due to suspicion DRE (19.7%), 117 due the simultaneous of both earlier (7.1%) and 302 with %fPSA level =15% and PSA between 2.5-3.9 ng/ml (18.3%). It were diagnosed 652 cases of PC (3.7%). Among them, 609 (93.4%) were clinically localized (T1- 2) and 43 (6.6%) were T3-4. In the image exams, 26 (4%) were N1 and 18 (2.8%) were M1. The comparison between both groups showed lower serum PSA values (p <0.001), more favorable clinical stage (p <0.001) and Gleason score in group I (p <0.001). CONCLUSIONS: The MCPU has proved to be an important tool in screening populations with poor medical access in a country with large territory and socioeconomic inequalities such as Brazil. The screening showed statistically significant improvement of clinical staging at diagnosis compared to patients diagnosed in the routine of the BCH

Introdução: O câncer de próstata metastático resistente à castração apresenta sobrevida inferior a 30% em cinco anos, e o único tratamento disponível para os pacientes até pouco tempo atrás era o docetaxel. Com o maior entendimento dos mecanismos de resistência desse câncer às terapias utilizadas, novas drogas foram desenvolvidas, entre elas a abiraterona, que atua no bloqueio da enzima 17-? desidrogenase-hidroxiesteroide e a enzalutamida que age diretamente nos receptores de androgênios. Foi realizada uma revisão sistemática dos estudos que avaliaram a eficácia e segurança da abiraterona mais prednisona e da enzalutamida em pacientes com CPMRC, tanto previamente quanto posteriormente ao tratamento com docetaxel. Métodos: A pesquisa bibliográfica foi realizada em março de 2016 nas bases da Pubmed, Google Acadêmico e ClinicalTrials.gov. Os seguintes termos de pesquisa foram utilizados \"prostate cancer metastatic resistent\", \"prostate cancer\", \"abiraterone\" e enzalutamide\", restringido a estudos clínicos randomizados fase III, duplo cego e multicêntrico, publicados nos idiomas inglês, espanhol. Os estudos selecionados foram analisados de forma independente por LRM e AAN. Os critérios para análise foram a sobrevida global (SG), sobrevida livre de progressão (SLP) e o tempo até a progressão do PSA (TP PSA), previamente e após o uso de docetaxel. Resultados: Foram identificados 53 estudos. Aplicando os critérios de inclusão foram selecionados quatro estudos que juntos somaram uma população de 5.199 (2.394 pacientes após docetaxel e 2.805 pacientes previamente ao uso do docetaxel). Em relação a SG, a abiraterona reduziu o risco de morte em 26% após docetaxel (HR 0,74 95% IC 0,64-0,86) e 19% no pré-docetaxel (HR 0,81 95% IC 0,70-0,93), e a enzalutamida reduziu em 37% após (HR 0,63 95% IC, 0,53-0,75) e 23% no pré-docetaxel (HR 0,77 95% IC 0,67-0,88). Em relação a SLPr, abiraterona retardou a progressão da doença em 34% após (HR 0,66 95% IC 0,58-0,76) e 48% na pré-quimio (HR 0,52 95% IC 0,45-0,61), e a enzalutamida em 60% após quimio (HR 0,40 95% IC 0,35-0,47) e 68% na pré-quimio (HR 0,32 95% IC 0,28-0,36). Abiraterona aumentou o tempo até a progressão do PSA em 64% após quimio (HR 0,36 95% IC 0,52-0,78) e 50% na pré-quimio (HR 0,50 95% IC 0,43- 0,58), e a enzalutamida em 75% após quimio (HR 0,25 95% IC 0,20-0,30) e 83% na pré- quimio (HR 0,17 95% IC 0,15-0,20). Conclusão: Tanto abiraterona quanto enzalutamida demonstraram melhores resultados no aumento da SG, da SLPr e do TP PSA no tratamento de pacientes com CPMRC. Não há um estudo comparativo direto e devido a diferenças nos critérios de inclusão e dos grupos comparadores entre esses estudos clínicos, não é possível uma análise indireta entre tais medicamentos. Assim, questões como qual a melhor estratégia de tratamento na pré e pós-quimio e o sequenciamento ideal para enfrentamento do CPMRC não podem ser respondidas. / Introduction: Castrate-resistant metastatic prostate cancer presents survival less than 30% in five years, and the only treatment available to patients until recently was docetaxel. With the greater understanding of the mechanisms of resistance of this cancer to the therapies used, new drugs have been developed, among them abiraterone, which acts in the blockade of the enzyme 17-? dehydrogenase-hydroxysteroid and the enzyme that acts directly on the androgen receptors. A systematic review was made of studies evaluating the efficacy and safety of abiraterone plus prednisone and of enzyme adduct in patients with CPMRC both before and after docetaxel treatment. Methods: The literature search was prepared in March 2016 based on Pubmed, Google Scholar and ClinicalTrials.gov. The following search terms were used \"prostate cancer metastatic resistant\", \"prostate cancer\", \"abiraterone\" and \"enzyme\", restricted to phase III, double blind and multicenter clinical trials published in English, Spanish, Italian and Portuguese. The selected studies were analyze by LRM and AAN. The criteria for analysis were overall survival (OS), progression-free survival (PSA) and time to PSA progression (PS PSA), before and after docetaxel use. Results: Fifty-three studies were identified and, according to the inclusion criteria, four studies were selected that together added a population of 5,199 (2,394 patients after docetaxel and 2,805 patients prior to docetaxel use). In relation to SG, abiraterone reduced the risk of death by 26% after docetaxel (HR 0.74 95% CI 0.64-0.86) and 19% in predocetaxel (HR 0.81 95% CI 0.70- 0.93), and enzyme reductase reduced by 37% (HR 0.63 95% CI, 0.53-0.75) and 23% in pre docetaxel (HR 0.77 95% CI 0.67-0, 88). In relation to SLPr abiraterone delayed disease progression by 34% (HR 0.66 95% CI 0.58-0.76) and 48% in pre-chemotherapy (HR 0.52 95% CI 0.45-0, 61), and enzyme-alpha 60% after chemo (HR 0.40 95% CI 0.35-0.47) and 68% in pre-chemo (HR 0.32 95% CI 0.28-0.36). Abiraterone increased the time until PSA progression by 64% after chemo (HR 0.36 95% CI 0.52-0.78) and 50% in pre-chemo (HR 0.50 95% CI 0.43-0, 58), and enzyme-alpha in 75% after chemo (HR 0.25 95% CI 0.20-0.30) and 83% in pre-chemo (HR 0.17 95% CI 0.15-0.20). Conclusion: Both abiraterone and enzalutamide showed better results in increasing SG, SLPr and TP PSA in the treatment of patients with CPMRC. There is no direct comparative study and, because of differences in inclusion criteria and comparator groups between these clinical studies, indirect analysis of such drugs is not possible. Thus, questions such as the best pre and post chemo treatment strategy and the optimal sequencing for CPMRC coping cannot be answered.

Introdução: A curva de aprendizado em prostatectomia radical laparoscópica (PRL) pode variar de 10 a 150 procedimentos. Nesta fase, observa-se o maior número de complicações perioperatórias e conversões, além de resultados oncológicos e funcionais precários. Neste estudo, foram comparadas duas séries iniciais de PRL, realizadas pelos acessos transperitoneal (PRLT) e extraperitoneal (PRLE). Objetivos: Comparar os resultados obtidos com a realização da PRL pelos acessos transperitoneal e extraperitoneal durante a curva de aprendizado, avaliando-se os resultados perioperatórios, oncológicos e funcionais. Pacientes e métodos: Procedeuse a uma análise comparativa retrospectiva entre os dados das primeiras 40 PRLT realizadas no Hospital Getúlio Vargas de Pernambuco (grupo 1) e os dados das primeiras 40 PRLE realizadas no Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (grupo 2). Resultados: Comparando-se as variáveis trans-operatórias dos grupos 1 e 2, observamos diferenças estatísticas na preservação dos feixes vásculonervosos (77,5% vs 90%; p=0,008), tempo cirúrgico total (175,0 min vs 267,6 min;p<0,001) e da perda sanguínea estimada (177,5 ml vs 292,4 ml; p<0,001). Duas complicações (5%) foram observadas no grupo 1 (sangramento e lesão retal) e quatro (10%) ocorreram no grupo 2 (sangramento-2, lesão retal e de bexiga). Conversão para procedimento aberto foi necessária em um caso em cada grupo (2,5%). No período pósoperatório, as principais diferenças estatisticamente significantes entre os grupos 1 e 2 foram observadas nas comparações do tempo de internamento, do tempo de uso de catéter uretral, no uso de opióides, na recorrência bioquímica, na taxa de continência urinária e no tempo médio de seguimento. Nenhuma diferença estatística foi observada na incidência de complicações precoces (17,5% vs 17,5%; p= 1,000), porém três complicações maiores foram observadas no grupo 1, levando ao óbito de um paciente neste grupo. Dentre as complicações pós-operatórias tardias, observou-se uma diferença estatística quando se comparou a taxa de complicações menores entre os grupos 1 e 2 (30% vs 15%; p=0,004). Na comparação dos resultados oncológicos entre os grupos 1 e 2, observou-se diferença estatística no número total de margens cirúrgicas positivas (MCP) (10,3% vs 32,5%; p=0,016) e no estadiamento patológico (pT2: 94,8% vs 70% e pT3: 5,2% vs 30%; p=0,005). Correlacionando-se o achado de MCP e estadiamento patológico, observou-se que a maioria das MCP no grupo 1 ocorreu no estadio pT2 (75%), ao passo que 77% das MCP no grupo 2 ocorreu no estadio pT3. Conclusões: O acesso transperitoneal mostrou-se mais eficiente que o acesso extraperitoneal para a realização da prostatectomia radical laparoscópica durante a curva de aprendizado, porém enfatizando que a taxa de complicações graves foi maior quando este acesso foi utilizado. / Introduction: The learning curve in laparoscopic radical prostatectomy (LRP) can vary from 10 to 150 procedures. This procedure can be done using the transperitoneal or the extraperitoneal approach. So far, there is no consensus about the best way to perform LRP, mainly during the initial phases of the LRP programs. Objectives: To analyze and compare the perioperative, oncological and functional results obtained with both approaches while performing LRP during the learning curve. Patients and Methods: Data of the first 40 transperitoneal LRP (Group 1) performed at Getúlio Vargas Hospital of Recife were compared with the first 40 extraperitoneal LRP (Group 2) performed at Clinics Hospital of State University of São Paulo. Results: On transoperative time, statistically significant difference were observed comparing groups 1 and 2 related to the preservation of the neurovascular bundles (77,5% x 90%; p=0,008), overall surgical time (175 min x 267,6 min; p<0,001) and estimated blood loss (177,5 ml x 292,4 ml; p<0,001). Two complications (5%) were observed in group 1 (bleeding and rectal injury), whereas four (10%) were seen in group 2 (bleeding- 5%, rectal and bladder injury). Open conversion occurred in one case (2,5%) in both groups. On postoperative time, statistical difference comparing the groups 1 and 2 were seen in the in-hospital time, indwelling catheter time, narcotic use, biochemical recurrence and mean follow-up time. No statistical difference was observed related to the incidence of early complications (17,5% vs 17,5%; p= 1,000), but three major complications occurred in group 1, leading to one death in this group. On late postoperative time, a statistical difference was observed in the incidence of minor complications (30% vs 15%; p=0,004). Comparing the oncological results between groups 1 and 2, statistical difference was observed in the incidence of positive surgical margins (10,3% vs 32,5%; p=0,016) and pathological stages (pT2: 94,8% vs 70% and pT3: 5,2% vs 30%; p=0,005). The majority of positive margins in group 1 occurred in pT2 (75%), while this observation was more prevalent in pT3 (77%) in group 2. Conclusions: The transperitoneal approach was more efficient than the extraperitoneal approach for performing laparoscopic radical prostatectomy during the learning curve, but major complications were commoner when this approach was adopted.

Over 4,000 flavonoids have been identified, and among these, many of them are known to possess cardioprotective, anti-inflammatory, antimicrobial, and antitumor effects. However, most of these properties have yet to be fully understood. In this study, extracts from Gnaphalium gracile, thought to possess a mixture of flavonoids, have been tested for cytotoxic activity on pancreatic (MiaPaca, Panc28), colon (HCT-116, Caco-2), and prostate (PC3, LNCaP), cancer cell lines. Polar extracts from the leaves of G. gracile have the most cytotoxic effect on these cancer cell lines, particularly the prostate cancer cell lines PC3 and LNCaP. Evidence suggests the extracts have antineoplastic effects on these cancer cells lines possibly due to differentiation status on pancreatic and colon cancer, but not prostate cancer. Cytotoxic activity is not dependent on tumorigenic potential. Further research is needed to identify the bioactive compounds within these extracts.

Human γδ T cells expressing Vγ2Vδ2 TCRs monitor foreign- and self-prenyl pyrophosphate metabolites in isoprenoid biosynthesis to mediate immunity to microbes and tumors. Vγ2Vδ2 cells have been used for adoptive cancer immunotherapy with some partial and complete remissions. Most trials have used continuous zoledronate exposure to expand Vγ2Vδ2 cells. Zoledronate inhibits farnesyl pyrophosphate synthase causing isopentenyl pyrophosphate to accumulate that then stimulates Vγ2Vδ2 cells. Because zoledronate exposure is toxic, we hypothesized that a short period of exposure would reduce T cell toxicity but still be sufficient for monocytes uptake. Supporting this hypothesis, pulse zoledronate exposure with IL-2 resulted in more uniform expansion of Vγ2Vδ2 cells with higher purity and cell numbers as compared with continuous exposure. These Vγ2Vδ2 cells also had higher levels of CD107a and perforin and slightly increased tumor cytotoxicity. Importantly, adoptive immunotherapy with Vγ2Vδ2 cells derived by pulse stimulation controlled human PC-3 prostate cancer cells in immunodeficient NSG mice significantly better than those derived by continuous stimulation. Pulse zoledronate stimulation of Vγ2Vδ2 cells with IL-15 also resulted in higher purity and cell numbers. Like with CD8 αβ T cells, IL-15 preserved early memory Vγ2Vδ2 T cell subsets better than IL-2. However, despite this fact, adoptive immunotherapy with Vγ2Vδ2 cells derived with IL-15 showed similar inhibition of PC-3 tumor growth as those derived with IL-2. Thus, pulse zoledronate stimulation maximizes the purity, quantity, and quality of expanded Vγ2Vδ2 cells. This simple modification to existing protocols would likely enhance the effectiveness of adoptively transferred Vγ2Vδ2 T cells.