Orientador: Francisco Eduardo Martinez / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-18T07:43:37Z (GMT). No. of bitstreams: 1 Fontanelli_BeatrizAparecidaFioruci_M.pdf: 2160007 bytes, checksum: 5e358e9b41de70f7fd7c801fae0e0378 (MD5) Previous issue date: 2011 / Resumo: A exposição ao etanol altera a concentração do retinol e do all-trans-ácido retinóico (atAR) em vários tecidos. Os retinóides, retinol e atAR, são importantes para a diferenciação e manutenção das células epiteliais da próstata. O atAR se liga aos receptores de ácido retinóico (RARa, ß e y) e a interação receptor/ligante com a sequência responsiva ao retinóide no DNA, levam à transcrição de genes alvos. Assim, o atAR exerce efeitos no crescimento celular, diferenciação e apoptose, sendo essencial no desenvolvimento e diferenciação de órgãos e tecidos. Nosso objetivo foi analisar o retinol, o ácido retinóico e seus receptores, bem como, o índice de proliferação celular e de apoptose no lobo dorsolateral da próstata de ratos adultos UCh. Os animais foram divididos em quatro grupos experimentais (n=10/grupo): UChA (ingestão voluntária de etanol a 10% (v/v); UChACo (controle - ausência de etanol); UChB (ingestão voluntária de etanol a 10% (v/v) e UChBCo (controle - ausência de etanol). Após 150 dias de experimentação, os animais foram eutanasiados por decapitação e o sangue do tronco e os lobos dorsolaterais das próstatas foram coletados e processados: (1) para análises da concentração do retinol e do atAR no plasma e na próstata por meio de HPLC; (2) e análises de microscopia de luz para a proliferação celular (Ki-67), apoptose (Tunel) e para os receptores de ácido retinóico, por meio dos anticorpos anti-RARa, -ß e -y. O consumo crônico de etanol diminuiu a concentração do retinol no plasma dos grupos UChB (consumo alto de etanol) e UChA (consumo baixo de etanol). A concentração do retinol foi ainda menor no plasma do grupo UChB comparado ao UChA. No entanto, a concentração do retinol no tecido prostático não teve diferença significativa entre os grupos. O atAR aumentou significativamente somente no plasma do grupo UChB. Na próstata, a concentração do atAR aumentou no grupo UChB, enquanto que no UChA não houve diferença estatística. O RAR? na próstata dorsal e lateral dos ratos UCh não foi alterada em função do consumo de etanol. Já os RARß e -? apresentaram aumento do sinal na próstata dorsal do grupo UChB. Não houve diferença no índice de proliferação celular e de apoptose nas próstatas dorsais e laterais dos grupos experimentais. Conclui-se que o etanol altera a concentração do retinol e do atAR no plasma. Essa alteração é diretamente proporcional à quantidade de etanol consumida. Já na próstata, o retinol não é alterado pelo etanol. O consumo alto de etanol altera a concentração do atAR na próstata dorsolateral e a expressão dos RAR ß e y na próstata dorsal. A alteração da expressão dos RAR pode aumentar a sensibilidade da próstata à ação do atAR. O etanol não altera a proliferação celular e a apoptose na próstata dorsal e lateral / Abstract: Ethanol exposure alters the concentration of retinol and all-trans retinoic acid (atAR) in several tissues. Retinoids (retinol and atAR) are essential for the differentiation and homeostasis of the prostate epithelial cells. atAR binds to retinoic acid receptors (RAR a, ß and ?) and the interaction receptor/ligand with the sequence responsive to retinoid into DNA lead to transactivation of target genes. Thus, atAR directly produces their effects on cell growth, differentiation and apoptosis. This study aimed to analyze the retinol and all-trans-retinoic acid concentrations and its atAR receptors as well as the cell proliferation and apoptosis index upon the dorsolateral prostate lobe of adult UCh rats. All animals were divided into four experimental groups (n = 10/group): UChA (10% ethanol (v / v) voluntary intake); UChACo (without ethanol consumption); UChB (10% ethanol (v / v) voluntary intake) and UChBCo (without ethanol consumption). After 150 days of experimentation, animals were sacrificed followed by decapitation and trunk blood and dorsolateral prostate lobes collected. Samples of plasma and prostate by concentration analysis of the retinol and atAR were processed for HPLC. The cell proliferation and apoptosis immunoreactivities were assessed by Ki-67 and Tunel, respectively, and nuclear receptors by anti-RAR a,-ß and-y. Chronic ethanol consumption reduced the concentration of plasma retinol in UChB (high ethanol intake) and UChA groups (low ethanol intake). The retinol concentration in plasma was even lower in UChB compared to UChA group. However, the retinol concentration in prostate tissue was not significantly different between the groups. Concentration of atAR increased in plasma of UChB group, and was 96% higher in the UChA group. The prostate, atAR increased in the UChB group, while in UChA group no statistical difference. There was no statistical difference in proliferation cell and apoptosis in the dorsal and lateral prostate lobes between the groups. The expression of RAR a in the dorsal and lateral prostate of UCh rats was not altered as a function of ethanol consumption. Already RAR ß and-y showed increased signal in the dorsal prostate UChB group. We conclude that ethanol alters the concentration of retinol and atAR in plasma. This change is directly proportional to the amount of ethanol consumed. In the prostate, retinol is not altered by ethanol. The high ethanol intake alters the concentration of atAR in dorsolateral prostate and the expression of RARß and RARy in the dorsal prostate. Alteration in expression of RAR can increase sensitivity to the action of the atAR in prostate. Ethanol does not alter cell proliferation and apoptosis in the dorsolateral prostate / Mestrado / Anatomia / Mestre em Biologia Celular e Estrutural

Orientador: Sebastião Roberto Taboga / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-10T18:00:13Z (GMT). No. of bitstreams: 1 Corradi_LaraSilvia_D.pdf: 6064748 bytes, checksum: 6595f8861a86f33fa6761af9a8f6b0a8 (MD5) Previous issue date: 2008 / Resumo: Os andrógenos têm papel central na biologia da próstata, mas os estrógenos também podem afetar o crescimento e a diferenciação desta glândula. Em tecidos específicos do corpo, a proporção entre andrógenos e estrógenos pode diferir significativamente daquela encontrada no plasma sanguíneo. As concentrações intracelulares desses esteróides nos tecidos alvos são reguladas pelo metabolismo local de hormônios, altamente dependente de enzimas específicas metabolizadoras de esteróides, como a 5a-redutase (5a-r) e a Aromatase (aro). Na próstata, a ação androgênica é acentuada devido à conversão de testosterona em dihidrotestosterona por ação da enzima 5a-r, enquanto que a aro é responsável pela aromatização da testosterona em estrógenos. Assim, a síntese local destes esteróides assume grande importância em doenças que acometem tecidos glandulares, como o câncer de próstata, onde níveis anormais de hormônios promovem o desenvolvimento e o aumento de ocorrência de malignidades. Desse modo, os papéis dos andrógenos e estrógenos ativados podem ainda ser melhor compreendidos na manutenção das interações homeostáticas epitélio-estromais prostáticas. Estudos com inibidores específicos destas enzimas vêm sendo desenvolvidos a fim de se tentar esclarecer os reais papéis enzimáticos na manutenção da fisiologia prostática, assim como no surgimento e desenvolvimento de doenças. De modo geral, após 30 dias consecutivos de inibição, simultânea ou não, das enzimas 5a-r e aro, respectivamente por ação da Finasterida e do Letrozol, a próstata de gerbilos jovens, adultos e velhos mostrou-se com o compartimento epitelial e o estromal alterados e reorganizados na tentativa de adaptar-se à nova condição hormonal induzida. Imediatamente após o término de administração das drogas, na fase inicial do período de pós-tratamentos, as concentrações de testosterona e estrógenos modificaram-se, as células epiteliais tiveram o padrão de atividade secretora alterada e, no estroma, a matriz extracelular ficou totalmente remodelada, além das células musculares lisas e dos fibroblastos fenotipicamente alterados. Na fase final do período de pós-tratamento, ficou evidente a tentativa da próstata em se normalizar morfologica e fisiologicamente, porém, em nenhuma das idades isso aconteceu de modo efetivo. O bloqueio da metabolização de hormônios esteróides na próstata pode ser uma ferramenta importante para o estudo da interação epitélio-estroma tanto na fisiologia normal, quanto nas doenças da próstata. Sugere-se portanto, que o desbalanço entre as concentrações de andrógenos e estrógenos provocados pela Finasterida e de Letrozol, juntos ou separados, alterou significativamente a interação epitélio-estroma. Todos os resultados obtidos parecem ser indicativos de importantes sinais do novo cenário hormonal intraprostático. A recuperação do equilíbrio entre as concentrações hormonais da próstata fica comprometida após o bloqueio enzimático, dando, portanto, às enzimas em estudo, status de crucial importância para o desenvolvimento e manutenção da próstata. O estabelecimento de modelos experimentais para o estudo das relações entre epitélio e estroma e o conhecimento dos componentes celulares e macromoleculares da próstata tornam-se instrumentos muito importantes para o entendimento do desenvolvimento, da estrutura e da fisiologia desta glândula / Abstract: Androgens have substantial role in the biology of the prostate, but estrogens also can affect the growth and differentiation of this gland. In specific tissues of the body, the ratio between androgens and estrogens can differ significantly from that found into plasma. The intracellular concentrations of these steroids in target tissues are mediated by a local hormone metabolism, by specifcs steroid-metabolizing enzymes, as the 5a-redutase (5a -r) and the aromatase (aro). In the prostate, the androgenic action is accented due to the conversion of testosterone in dihydrotestosterone by the activity of the 5a -r, while aro enzyme is an alternative pathway for the aromatization of testosterone into estrogens. These locally syntheses of steroids hormones assume thus, a great importance to the prostate cancer, where abnormal hormone levels can promote development and proliferation of malignancy to this gland. The activated functions of androgens and estrogens may be better understood focusing the maintenance of homeostatic prostatic epithelial-stromal interactions. Studies conducted with inhibitors of these specific enzymes have been carried out in an effort to clarify the real role of steroid-metabolizing enzymes in the maintenance of prostatic physiology, as well as into malignant progression prostatic cancer. After 30 consecutive days of inhibition, simultaneous or not, of the enzymes 5a -r and aro, respectively by Finasterida and Letrozol, the prostate of young, adult and old gerbils revealed that epithelial and estromal compartments were totally modified and reorganized in the attempt to adapt it to the new induced hormonal condition. Immediately after the end of drugs administration, in the early phase of the post-treatments period, the concentrations of testosterone and estrogens had been altered, the epithelial cells had their secretore activity pattern modified and, in the stromal compartment, the extracellular matrix was totally remodeled. The smooth muscle cells and fibroblasts became fenotipically modified. In the late phase of the post-treatments period, it was evident the attempt of the prostate gland in became morphologically and physiologically as a normal gland, however, this was not achieved in any analyzed gerbil. These steroids hormone metabolization blockade seems to be a good tool for the study of the epithelium-stroma interaction both in normal and abnormal prostate gland. Based on this data, it is suggested that the unbalance between androgens and estrogens intraprostatic concentrations provoked by Finasteride and Letrozol, together or not, interfered into the prostatic compartments interactions significantly. The results seem to be indicative of important signals of the new locally hormonal scene within the prostate gland. The recovery of balance hormonal concentrations of the prostate is damaged after the enzymatic blockade what gives, therefore, to this to 5a -r and aro enzymes, a status of crucial key for the normal and abnormal prostate gland. The establishment of experimental models for this kind of study and the knowledge of the cellular and macromolecular components of the prostate are also fundamental for the agreement of development, structure and physiology of prostate / Doutorado / Biologia Celular / Doutor em Biologia Celular e Estrutural

Orientador: Rejane Maira Góes / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-23T14:41:50Z (GMT). No. of bitstreams: 1 Jesus_MarianaMarcielode_M.pdf: 3839823 bytes, checksum: 5cdcc794169929d299d222355a85e2d1 (MD5) Previous issue date: 2013 / Resumo: O resumo poderá ser visualizado no texto completo da tese digital / Abstract: The abstract is available with the full electronic document / Mestrado / Biologia Celular / Mestra em Biologia Celular e Estrutural

Orientador: Valéria Helena Alves Cagnon Quitete / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-18T10:44:47Z (GMT). No. of bitstreams: 1 Montico_Fabio_M.pdf: 7570263 bytes, checksum: a775095a6141dbe3b18c6ec768585813 (MD5) Previous issue date: 2011 / Resumo: A reposição androgênica representa uma alternativa para minimizar os efeitos prejudiciais do desequilíbrio hormonal em homens senis, embora seus efeitos sobre o desenvolvimento de doenças prostáticas ainda seja assunto controvertido. Assim, o objetivo deste estudo foi caracterizar aspectos estruturais e moleculares do lobo ventral da próstata de ratos senis frente à reposição de hormônios esteróides, relacionando as alterações decorrentes da terapia hormonal a possíveis condições de lesões prostáticas. Ratos machos (Sprague- Dawley) foram divididos em um grupo Jovem (JOV) (4 meses), que recebeu óleo de amendoim (5 mL/Kg, s.c.), e grupos senis (10 meses), submetidos aos tratamentos: grupo Senil (SEN): óleo de amendoim (5 mL/Kg, s.c.); grupo Testosterona (TEST): cipionato de testosterona (5 mg/Kg, s.c.); grupo Estrógeno (EST): 17?-estradiol (25 ?g/Kg, s.c.); grupo Castrado (CAS): castração cirúrgica; grupo Castrado-Testosterona (CT): castração e após 30 dias tratamento similar ao grupo TEST; grupo Castrado-Estrógeno (CE): castração e após 30 dias tratamento similar ao grupo EST. Após 30 dias de tratamento, foram coletadas amostras de sangue, para dosagens hormonais séricas, e do lobo ventral, para análises em microscopias de luz e eletrônica de transmissão, morfométricas, imunohistoquímicas e Western Blotting. As moléculas investigadas foram: distroglicanas ? e ? (?-DG e ?-DG), receptor do fator de crescimento homólogo à insulina tipo I (IGFR-1), metaloproteinase-9 de matriz (MMP-9), fator de crescimento do endotélio vascular (VEGF) e endostatina. Redução dos níveis séricos de testosterona foi verificada na senescência, com aumento destes após a reposição hormonal no grupo TEST. O estroma do grupo SEN apresentou hipertrofia e células inflamatórias. Após a reposição hormonal na senescência ou frente à castração, verificou-se atrofia no epitélio, células epiteliais com halo citoplasmático claro ao redor do núcleo, microácinos e manutenção do estroma hipertrófico com células inflamatórias. Diminuição dos níveis das DGs foi verificada na senescência, sendo que após a terapia hormonal ocorreu aumento dos níveis protéicos dessas moléculas, especialmente nos grupos que receberam estradiol. Aumento do IGFR-1 e da MMP-9, bem como distribuição diferencial dessas moléculas no compartimento epitelial, foram observados nos grupos submetidos à reposição hormonal e no grupo CAS. O grupo SEN caracterizou acréscimo dos níveis de VEGF, sendo o inverso observado para a endostatina. A terapia hormonal e a castração levaram à elevação dos níveis de VEGF, sobretudo nos grupos EST, CAS e CE. Em oposição, a endostatina demonstrou-se aumentada especialmente nos grupos submetidos à reposição de testosterona. Os presentes resultados sugeriram que o desequilíbrio entre andrógenos e estrógenos verificado na senescência foi acentuado após a terapia hormonal e a castração, potencializando as alterações estruturais associadas a esse período e rompendo o equilíbrio das sinalizações parácrinas prostáticas, com aumento simultâneo de IGFR-1 e MMP-9 e geração de fatores pró e anti-angiogênicos em resposta ao tratamento com estrógenos e andrógenos, respectivamente. Assim, concluiu-se que a terapia hormonal, apesar de seus efeitos positivos sobre as DGs, gerou microambiente prostático reativo, caracterizado por aumento de um fator mitogênico e da remodelação tecidual bem como por desequilíbrio da angiogênese, o que possivelmente comprometeu a função do órgão e o predispôs a desordens glandulares / Abstract: Androgen replacement is an alternative to minimize the harmful effects of hormonal imbalance in elderly men, even though its influence on the development of prostatic diseases is unclear. Thus, the aim herewith was to characterize structural and molecular features of the ventral prostate of senile rats submitted to steroid hormone replacement, relating the alterations resulting from hormonal therapy to possible prostatic lesions. Male rats (Sprague-Dawley) were divided into Young group (YNG) (4 months old rats), which received peanut oil (5 mL/Kg, s.c.), and senile groups (10 months old rats), submitted to the treatments: Senile group (SEN): peanut oil (5 mL/Kg, s.c.); Testosterone group (TEST): testosterone cipionate (5 mg/Kg, s.c.); Estrogen group (EST): 17?-estradiol (25 ?g/Kg, s.c.); Castrated group (CAS): surgical castration; Castrated-Testosterone group (CT): castration and after 30 days treatment similar to TEST group; Castrated-Estrogen group (CE): castration and after 30 days treatment similar to EST group. After 30 days treatment, blood samples were collected for hormonal analysis and ventral prostate samples were processed for light and transmission electronic microscopies, morphometric, immunohistochemical and Western Blotting analysis. The investigated molecules were alfa and beta dystroglycans (?DG and ?DG), insulin-like growth factor receptor-1 (IGFR-1), matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF) and endostatin. Decreased serum testosterone levels were verified in senescence, with an increase after hormonal replacement in the TEST group. Hypertrophied stroma with inflammatory cells was seen in the SEN group. After hormonal therapy in the senescence or following castration, atrophic and pale cytoplasmatic epithelial cells, microacini and hypertrophied stroma were observed. Decreased DG levels were verified in senescence, but there was an increase of these levels following hormonal therapy, especially in the groups treated with estradiol. Increased IGFR- 1 and MMP-9 protein levels and differential distribution of these molecules were observed in epithelial compartment in those groups which received hormone replacement and in the CAS group. SEN group showed increased VEGF levels in contrast to decreased endostatin levels. Hormonal therapy and castration led to raised VEGF levels, mainly in EST, CAS and CE groups. On the other hand, endostatin was increased especially in those groups submitted to testosterone replacement. The present results suggested that the imbalance between androgens and estrogens in senescence was enhanced after hormone replacement and castration, intensifying structural changes associated with this period. Also, there was a disruption of prostatic paracrine signaling balance, with simultaneous increase of IGFR-1 and MMP-9 and generation of pro and anti-angiogenic factors in response to treatment with estrogens and androgens, respectively. Thus, it could be concluded that despite its positive effects on DGs levels, hormonal therapy created a reactive prostatic microenvironment, characterized by increase of a mitogenic factor and tissue remodelling as well as prostatic angiogenesis imbalance, which could compromise glandular functions and lead to the emergence of prostatic lesions / Mestrado / Anatomia / Mestre em Biologia Celular e Estrutural

Orientador: José Wilson Magalhães Bassani / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de Computação / Made available in DSpace on 2018-08-26T12:41:31Z (GMT). No. of bitstreams: 1 Almeida_JoaoCarlosMartinsde_M.pdf: 13918269 bytes, checksum: 78378428b79af0919ecab395a454bf98 (MD5) Previous issue date: 2014 / Resumo: O estudo urodinâmico tem importante papel na determinação da necessidade de pacientes portadores de sintomas do trato urinário inferior (STUI) se submeterem à cirurgia da próstata. O estudo pressão-fluxo pode contribuir muito para a quantificação dos STUI, porém é invasivo, demorado e de custo elevado. Métodos alternativos têm sido desenvolvidos procurando minimizar o procedimento invasivo necessário para medição da pressão vesical. Um método alternativo foi desenvolvido no Centro de Engenharia Biomédica da UNICAMP e consiste num dispositivo denominado conector uretral (Pat. N. PI 0502171-5). Trata-se de um dispositivo cilíndrico e vazado, que permite o escoamento de urina por seu interior. O paciente é orientado a introduzir o conector na uretra antes de iniciar a micção e, ao urinar, solicitado a ocluir a saída do dispositivo, interrompendo o fluxo urinário por um curto intervalo e permitindo o registro da pressão dentro da bexiga urinária. Neste trabalho, uma instrumentação e uma nova versão do conector uretral foram desenvolvidas para medir a pressão vesical em homens. Aplicado em testes clínicos em indivíduos com sintomas de obstrução infravesical decorrente de hiperplasia benigna da próstata, o conjunto demonstrou capacidade de diferenciar indivíduos com obstrução de indivíduos não obstruídos sem diferença significativa do que se obtém no estudo pressão-fluxo (p > 0,05). O conector apresentou sensibilidade e especificidade de 66,7%, valores similares aos dos métodos alternativos descritos na literatura. Além disso, 88,5% dos indivíduos relataram preferir o conector uretral ao estudo pressão-fluxo, indicando a importância do conector como método alternativo minimamente invasivo ao estudo urodinâmico convencional / Abstract: Urodynamic assessment has an important role in the decision of patients complaining of lower urinary tract symptoms (LUTS) to undergo prostate surgery. Pressure-flow study may contribute to the evaluation of LUTS, but its procedure is invasive, time-consuming and expensive. Alternative methods have been developed aiming at measuring vesical pressure in a less invasive manner. An alternative method was developed in the Center for Biomedical Engineering at UNICAMP and consists of a device named urethral connector (Pat. N. PI 0502171-5). The device has a cylindrical shape and an internal longitudinal hole that permits urine to pass through it. The patient is instructed to insert the urethral connector into his urethra before starting to urinate. During micturition, he is instructed to occlude the urine outlet of the device for a short period of time so that vesical pressure may be measured. In this work, an instrumentation and a new version of the urethral connector were developed to measure vesical pressure in men. The device and instrumentation were applied in clinical tests in individuals reporting LUTS secondary to infravesical obstruction due to benign prostatic hyperplasia. It was shown that this new method is capable of differentiating obstructed individuals from nonobstructed ones. Results were not significantly different from those obtained when conventional pressure-flow study was applied (p > 0.05). The urethral connector had a sensibility and specificity of 66.7%, values similar to those found by the alternative methods described in the literature. About 88.5% of the individuals evaluated reported to prefer the new method instead of the conventional assessment which reinforces the importance of the urethral connector as a less invasive and alternative method to the conventional urodynamic study / Mestrado / Engenharia Biomedica / Mestre em Engenharia Elétrica

Orientador: Miriam Dambros Lorenzetti / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-28T00:11:28Z (GMT). No. of bitstreams: 1 Souza_FelipeGoncalvesSchrodere_M.pdf: 1431848 bytes, checksum: df85a182625e0c75b33faa908d302d6c (MD5) Previous issue date: 2015 / Resumo: Introdução: A neoplasia de próstata é a segunda mais prevalente entre os homens, com estimativa de cerca de 68 mil casos novos no Brasil em 2014. Além disso, é a segunda causa de morte no sexo masculino por neoplasias. O câncer de próstata raramente causa sintomas em seus estágios iniciais, por isso existe a necessidade do diagnóstico precoce. Atualmente a forma de rastreamento desta neoplasia ainda é controverso, porém quando realizado, baseia-se em exame digital retal da próstata e na mensuração sérica dos níveis do PSA. Estudos de rastreamento populacional mostraram uma diminuição de 41% no casos de neoplasia avançada de próstata e risco de diagnóstico de câncer 46% maior, quando comparados a grupos não submetidos a rastreamento. O PSA não é câncer específico, então, os refinamentos do PSA (relação do PSA livre/total, densidade do PSA e velocidade do PSA) aparecem como métodos para melhorar sua especificidade, na tentativa de diminuir o número de biópsias desnecessárias. Quando existe suspeita diagnóstica, é indicada uma biópsia guiada por ultrassonografia transretal, procedimento que não é isento de complicações, principalmente as infecciosas. Objetivos: Avaliar a positividade para adenocarcinoma de próstata em biópsias guiadas por ultrassonografia transretal, e estratificá-la de acordo com a idade, com o valor do PSA total, com a densidade do PSA e com a relação entre a fração livre do PSA e o PSA total. Materiais e Métodos: Analisamos retrospectivamente os resultados obtidos no serviço de urologia do Hospital Municipal Dr. Mário Gatti com relação à positividade das biópsias estratificadas pela idade, PSA total, percentual da fração livre do PSA e densidade do PSA, comparando-os com os dados descritos na literatura. Resultados: Foram realizadas 314 biópsias no período de janeiro de 2011 à novembro de 2012. A média de idade dos pacientes foi de 65,2 (± 8,32) anos e a positividade foi de 44,9%. A positividade das biópsias foi maior com o aumento da idade (p<0,001), com o aumento do PSA (p<0,001), com o aumento da densidade (p<0,001) e com a diminuição da relação do PSA (p=0,002) Conclusão: A neoplasia prostática correlacionou-se significativamente com o aumento da idade, do PSA total, da densidade do PSA, e com a diminuição da relação entre o PSA livre sobre o PSA total / Abstract: Introduction: Prostate neoplasia is the second most prevalent neoplasia in men and about 68 thousand new cases were estimated in 2014 in Brazil. In addition, it is the second most common cause of cancer related death in men. Prostate cancer rarely causes symptoms at an early stage, hence the need of an early diagnosis. Although there is still no consensus about how to screen this neoplasia, it is done through digital rectal examination and measurement of PSA levels. Population screening trials showed a decrease of 41% in new cases of advanced prostate neoplasia. The risk of a cancer diagnosis increased 46% when compared to the group who was not screened. PSA is not cancer specific. Therefore, PSA features (relation between free PSA and total PSA, PSA density, PSA velocity) are used to increase its specificity, attempting to reduce the number of unnecessary biopsies. Once there is a cancer suspicion, a biopsy guided by transrectal ultrasonography is indicated, which is not a complication free procedure, mainly infection. Objective: to assess the positiveness of prostatic adenocarcinoma in transrectal ultrasonography guided biopsies and to evaluate it according to age, total PSA value, PSA density, and the relation between free PSA and total PSA. Materials and Methods: This is a retrospective study with patients who underwent prostatic biopsy guided by transrectal ultrasonography done by the urology team from Hospital Municipal Dr. Mário Gatti. Collected data included patient¿s age, total PSA value, PSA density and the relation between free PSA and total PSA. Results: 314 prostatic biopsies guided by transrectal ultrasonography were analyzed between January 2011 and November 2012. Patient¿s mean age was 65.2 (+/-8.32) years. A positive biopsy to adenocarcinoma was found in 44.9% of the patients. The number of positive biopsies was higher among older patients (p<0.001). It was also higher the higher the PSA (p<0.001) and the higher the PSA density (p<0.001). It was inversely related to PSA relation (p=0.002). Conclusion: There is a direct correlation between prostatic neoplasia and age, value of PSA and PSA density. Additionally, prostatic neoplasia is inversely proportional to the relation of free PSA over total PSA / Mestrado / Fisiopatologia Cirúrgica / Mestre em Ciências da Cirurgia

Prostata-adenocarcinom är den vanligaste cancerformen hos män. Anti-PSA och anti-P501S är frekvent använda antikroppar för diagnostisering av prostata-adenocarcinom i vävnadsbiopsier, men kan ha svag eller negativ infärgning av antigenen. Antikroppen anti-NKX3.1 har visat sig ha en högre sensitivitet och starkare infärgning än anti-PSA och anti-P501S vid prostata-adenocarcinom. Det rekommenderas att använda anti-NKX3.1 tillsammans med anti-PSA och/eller anti-P501S för att diagnostisera metastaserande prostata-adenocarcinom. Studiens syfte var att utvärdera och implementera antikroppen anti-NKX3.1 för diagnostisering av metastaserande prostata-adenocarcinom. Positiva vävnadskontroller (prostatavävnad och testisvävnad) och negativ vävnadskontroll (appendix) färgades med anti-NKX3.1 i Roche Ventana Benchmark ULTRA med immunohistokemisk färgning. Förbehandlingarna CC1 mild, CC1 standard och Proteas 1 samt visualiseringskiten OptiView samt UltraView jämfördes. Spädningen optimerades från 1:50-1:200. Vävnadssnitten graderades sedan från 1 (ingen infärgning)-6 (starkast infärgning). Visualiseringskitet OptiView samt förbehandlingen CC1 standard gav den starkaste infärgningen med anti-NKX3.1 och valdes som det optimala visualiseringskitet respektive förbehandlingen. Med tanke på materialkostnad och smidighet valdes spädningen 1:100 som den optimala spädningen. / Prostate adenocarcinoma is the most common form of cancer in men. Anti-PSA and anti-P501S are frequently used antibodies for diagnosing of prostate adenocarcinoma in tissue biopsies but may show weak or negative staining of the antigens. Antibody anti-NKX3.1 has been shown to have a higher sensitivity and stronger staining then anti-PSA and anti-P501S in prostate adenocarcinoma. It is recommended to use anti-NKX3.1 along with anti-PSA and/or anti-P501S to diagnose metastases of prostate adenocarcinoma. The purpose of the study was to evaluate and implement the antibody anti-NKX3.1 for diagnosis of metastatic prostate adenocarcinoma. Positive tissue controls (prostate and testis) and negative tissue control (appendix) were stained with anti-NKX3.1 immunohistochemical staining in Roche Ventana Benchmark ULTRA machine. The pre-treatments CC1 mild, CC1 standard and Protease 1 and the visualization kits OptiView and UltraView were compared. The dilution was optimized from 1:50-1:200. Immunohistochemical staining of the tissue was graded from 1 (no staining) - 6 (strongest staining). Visualization kit OptiView and pre-treatment CC1 standard gave the strongest staining with anti-NKX3.1 and were chosen as the optimal visualization kit and pre-treatment. Considering the material cost and flexibility, the 1:100 dilution was chosen as the optimal dilution.

Das Prostatakarzinom ist die häufigste Krebserkrankung des Mannes. Bei den zu Tode führenden Tumoren wird es im Jahre 2003 nach dem Bronchialkarzinom an 2. Stelle stehen. Diese Inzidenz zeigt, dass dringend neue diagnostische Marker und therapeutische Zielgene zur Behandlung von Prostatakrebs benötigt werden. Ziel dieser Dissertation war es, mit Hilfe der DNA-Chiptechnologie neue tumorrelevante Gene für eine Small-Molecule- und Antikörper-Basierte Therapie des Prostatakarzinoms zu identifizieren. Auf einen proprietären Tumor-Chip der Firma metaGen Pharmaceuticals GmbH wurde mikrodissektiertes Normal- und korrespondierendes Tumorgewebe von 52 Prostatatumorpatienten hybridisiert. Mit Hilfe bioinformatischer Analysen der Chipergebnisse konnte das Gen TRPM8 identifiziert werden, das in Prostatatumoren in mehr als 56% der Patienten überexprimiert ist. Northern-Blot, Dot-Blot und Chipexperimente zeigten, dass TRPM8 ungewöhnlich gewebespezifisch exprimiert wird. In mehr als 400 getesteten Tumorpatienten und in 23 Normalgeweben wurde TRPM8 ausschließlich in der Prostata und neuroendokrinen Tumoren nachgewiesen. TRPM8 gehört zur Familie der Transient Receptor Potential Channel Proteins. Es konnte hier erstmals in Fluoreszenz-Resonanz-Energie-Transfer-Experimenten (FRET) gezeigt werden, dass TRPM8 Multi-Homomere bildet. Dies wurde bisher nur für Kanäle anderer TRP-Subfamilien (TRPV und TRPC) gezeigt. Weiterhin konnten erstmals mehrere Spleißvarianten von TRPM8 identifiziert werden. Quantitative RT-PCR Experimente zeigten, dass diese noch stärker in Prostatatumoren überexprimiert sind als TRPM8 selbst. Des Weiteren wurde ein neues Gen auf dem DNA-Gegenstrang von TRPM8 entdeckt, das mit Exon 11 von TRPM8 100% komplementär ist und an der Regulation von TRPM8 beteiligt sein könnte. Der Promotor von TRPM8 wurde durch eine in silico Analyse identifiziert und in vitro bestätigt. Obwohl eine starke androgenabhängige Expression von TRPM8 in LNCaP Zellen gezeigt werden konnte, wurden keine Bindungsstellen für androgenabhänginge Elemente gefunden. Allerdings ließen sich drei Bindungsstellen des androgenregulierten Homeoboxgens NKX3.1 identifizieren. Die Ergebnisse dieser Arbeit zeigen, dass TRPM8 und seine Isoformen aufgrund ihrer Gewebspezifität ausgezeichnete Angriffspunkte für eine zielgerichtete Prostatakrebstherapie sind. / Prostate cancer is the most commonly diagnosed malignancy in men in the Western World. In 2003 malignancies of the prostate will be the second most common fatal cancer in men after lung cancer as estimated by the American Cancer Society. Despite the tremendous efforts made in the past to improve the treatment of prostate cancer patients, there is still an urgent need for new markers and therapeutic targets for medication. The aim of this thesis was the identification of new genes relevant in prostate cancer, which could be used in a small-molecule or antibody based therapy of prostate cancers. Microdissected matched prostate cancer and normal tissues of 52 prostate cancer patients were hybridized to a proprietary high density Cancer-Chip based on Affymetrix GeneChip technology. Using a bioinformatic analysis, it was possible to identify TRPM8, which was highly overexpressed in 56% of prostate cancer patients. Northern blot, dot blot and gene chip experiments revealed that TRPM8 expression is extremely tissue specific. Of 400 patients and 23 tissues tested, TRPM8 expression could only be detected in the prostate and neuroendocrine tumors. Functionally, the protein belongs to the transient receptor potential channel family of non-voltage gated proteins. It could be shown for the fist time that TRPM8 subunits form homomers using FRET technology. Molecular characterization of TRPM8 transcription revealed multiple splice forms of TRPM8. Further, it was possible to identify a new mRNA present on the opposite strand of TRPM8, which was 100% complementary to exon 11 of TRPM8, thus it could possibly function as a regulatory RNA of TRP channel. All of these isoforms were found to be even higher overexpressed in prostate tumors than TRPM8 itself. The promoter region of TRPM8 was identified using in silico methods and confirmed in promoter reporter assays. Although a high androgen dependent transcriptional activation of TRPM8 could be found by RT-PCR in LNCaP cells, no androgen responsive elements was identifiable within the promoter region. On the other hand three binding sites for the androgen dependent homeobox gene NKX3.1 and several other homeobox genes were discovered. The results of the thesis show that TRPM8 and its isoforms are, due to their tissue specificity, ideal targets for the development of new therapeutic drugs for the treatment of prostate cancer.

Ziele: Trotz ihrer signifikanten Morbidität stellt die transurethrale Resektion der Prostata (TURP) den gold standard dar für die instrumentelle Therapie der durch die benigne Prostatahyperplasie (BPH) bedingten Blasenauslassobstruktion. Mit dem Hochleistungs-Holmium: YAG Laser kann endoskopisch, in einer relativ blutungsarmen Weise, Prostatagewebe enukleiert werden. Wir verglichen die Technik der transurethralen Holmium-Laser-Enukleation der Prostata (HoLEP) mit der Standard-TURP bei der operativen Therapie von Prostataadenomen. Wir präsentieren die perioperativen Daten, die Kurz- und Langzeitergebnisse dieser randomisierten klinischen Studie. Material und Methoden: Insgesamt wurden 200 Patienten mit einer urodynamisch nachgewiesenen benignen Prostataobstruktion bei einem Prostatavolumen kleiner 100g im transrektalen Ultraschall in ein TURP- und HoLEP-Kollektiv randomisiert. Alle Patienten wurden präoperativ und postoperativ nach 1,6, 12, 18, 24 und 36 Monaten durch Erhebung des American Urological Association symptom score und Messung der maximalen Harnflussrate und des Restharns evaluiert. Die perioperativen Daten und die postoperativen Ergebnisse wurden verglichen. Alle Komplikationen wurden vermerkt. Ergebnisse: Bei der HoLEP waren die Katheterzeit, der Krankenhausaufenthalt und der Hämoglobinverlust signifikant geringer, jedoch die Operationszeit signifikant länger als bei der TURP. HoLEP und TURP führten zu signifikanter und anhaltender Verbesserung aller Miktionsparameter, wobei der Restharn bis einschließlich 36 Monate postoperativ in der HoLEP Gruppe signifikant geringer blieb. Auch die perioperative Morbidität war bei der HoLEP kleiner; Kontinenz, Potenz und Spätkomplikation waren in beiden Gruppen ähnlich. Schlussfolgerungen: HoLEP ist mindestens genauso effektiv wie TURP. HoLEP dauert länger, jedoch sind die perioperative Morbidität, Katheter- und Krankenhausdauer geringer. Beide Methoden zeigen befriedigende Langzeitergebnisse mit wenigen Spätkomplikationen. Dies bestätigt die Dauerhaftigkeit des Therapieerfolges dieser 2. Verfahren. / Purpose: Transurethral electrocautery resection (TURP) is generally regarded as the gold standard surgical treatment for bladder outflow obstruction due to benign prostatic hyperplasia (BPH) despite its rather high morbidity. The high powered holmium: YAG laser can be used endoscopically to enucleate prostatic tissue in a relatively bloodless manner. The technique of transurethral holmium laser enucleation of the prostate (HoLEP) was compared to standard TURP for the surgical management of prostate adenomas. We present the perioperative data, short and long-term results of this randomized clinical trial. Materials and Methods: A total of 200 patients with urodynamic obstruction with a prostate less than 100 gm on transrectal ultrasound were randomized into 2 comparable groups and assigned to HoLEP or TURP. All patients were assessed preoperatively and followed prospectively 1, 6, 12, 18, 24, and 36 months postoperatively with an American Urological Association symptom score, peak urinary flow rate and post-void residual measurement. Perioperative data and postoperative outcome were compared. All complications were noted. Results: HoLEP was significantly superior to TURP in terms of catheter time, hospital stay and hemoglobin loss but operative time was significantly longer. HoLEP and TURP resulted in a significant and lasting improvement of all parameters with post-void residual volume still significantly better in the HoLEP group at the 36-month assessment. Perioperative morbidity was less in the HoLEP group; continence, potency and late complications were similar in both groups. Conclusion: HoLEP is at least as effective as TURP. HoLEP takes longer than TURP, but perioperative morbidtity, catheter time and hospital stay is less. Both treatments have satisfactory long term results with a low late complication rate. This confirms the durability of these 2 procedures.

Prostatacancer är en av världens vanligaste cancerformer. Varje år diagnostiseras ungefär 1,3 miljoner män världen över med prostatacancer och trots det vet man inte de bakomliggande orsakerna. Det finns flera studier som visar att det finns en koppling mellan infektion och olika typer av cancer och Cutibacterium acnes återfinns i hög utsträckning i prostatacancervävnad. Det har därför föreslagits att det finns ett samband mellan infektion av C. acnes och prostatacancer. Närvaro av tumörassocierade makrofager har visats sig vara gynnande för olika typer av cancer och från dessa makrofager frisätts kemokiner, bland annat CCL22. CCL22 kan vara inblandad i en lokal hämning av immunförsvaret som ofta förknippas med tillväxt av cancer. I detta arbete odlas makrofager från blodgivare med C. acnes för att ta reda på om makrofagerna ökar sitt uttryck av CCL22 mRNA. Genom att analysera resultaten från en kvantitativ realtids-PCR indikeras det att makrofager som behandlats med C. acnes signifikant ökar sitt uttryck av CCL22 mRNA. Sammanfattningsvis bedöms resultaten från detta arbete styrka uppfattningen att det kan finnas en koppling mellan C. acnes och orsakerna bakom prostatacancer men mer arbete återstår för att kunna klargöra dess relevans. / Prostate cancer is one of world´s most common forms of cancer. Every year about 1.3 million men around the world are diagnosed with prostate cancer and even so we cannot fully explain the etiology. There have been several studies indicating that there is a correlation between infection and different forms of cancer. Cutibacterium acnes (C.acnes) can be found to a large extent in prostate cancer tissue and a correlation between infection of C. acnes and prostate cancer has therefore been suggested. The presence of tumor-associated macrophages has been shown to favor various types of cancer. Several chemokines including CCL22 are released from these macrophages. CCL22 may be involved in a local inhibition of the immune system that is often associated with cancer growth. In this work, macrophages from a blood donor are grown with C. acnes to find out if the macrophages increase their expression of CCL22 mRNA after this exposure. By analyzing the results of a quantitative real-time PCR for CCL22 mRNA, it was demonstrated that macrophages treated with C. acnes significantly increase their expression of CCL22 mRNA. In conclusion, the results of this work indicate that there could be a link between C. acnes and the causes of prostate cancer, but more work remains to be able to clarify its relevance.