Thesis (doctoral)--Rijksuniversiteit te Utrecht. / "Appendix" (p. 45-76) inserted in pocket at end.

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 87-114).

Thesis (doctoral)--Rijksuniversiteit te Utrecht. / "Appendix" (p. 45-76) inserted in pocket at end.

Thesis (Ph. D.)--University of Hong Kong, 2005. / Title proper from title frame. Also available in printed format.

Orientador: Maria Aparecida Custodio Domingues / Resumo: As neoplasias testiculares constituem aproximadamente 1% dos cânceres masculinos. Dentre eles, os tumores que têm origem nas células germinativas (TTCG) são os mais frequentes. Por sua natureza, esses tumores têm padrão morfológico variado, sendo distribuídos em dois subgrupos: os seminomas e os não-seminomas. Esses últimos, por sua vez, são classificados em carcinomas embrionários, tumores do seio endodérmico, coriocarcinomas, teratomas e tumores mistos de células germinativas. Há consenso na literatura de que a invasão vascular verificada histologicamente no tumor primário é o melhor indicador preditivo de progressão da doença e recidiva. Estudos recentes demonstram o papel dos fatores de crescimento e seus receptores para avaliação prognóstica dos TTCG, no que o uso da imuno-histoquímica é fundamental. O presente estudo procurou avaliar pela técnica imuno-histoquímica à ocorrência da expressão do receptor do fator de crescimento epidérmico (EGFR), do Her2 e do c-Kit em uma série de TTCG não-seminomas primários de testículo. Além disso, explorou a possível relação entre a expressão desses marcadores com a evolução dos pacientes após terapêutica convencional. A série foi constituída de amostras parafinadas existentes no Laboratório de Anatomia Patológica e Citologia do Hospital Amaral Carvalho de Jaú, totalizando 103 pacientes que receberam o diagnóstico de tumor testicular de células germinativas não-seminomas (TTCGNS) no período 1996-2010. Dentre os 103 casos, predominaram... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Testicular neoplasms constitute approximately 1% of male cancers. Among these, germ cell tumors (TGCT) are the most prevalent. These tumors show varied morphological patterns and are distributed into two subgroups: seminomas and non-seminomas. The latter, in turn, are classified as embryonal carcinomas, endodermal sinus tumors, choriocarcinomas, teratomas and mixed germ cell tumors. The consensus in the literature is that histological verification of vascular invasion (VI) in the primary tumor is the best predictor of disease progression and recurrence. However, recent studies have highlighted the role of growth factors and their receptors for prognostic evaluation of TGCT, in which the use of immunohistochemistry is fundamental. This study sought to evaluate the occurrence of epidermal growth factor receptor (EGFR), Her2 and c-Kit in a series of primary non-seminoma TGCTs. In addition, possible relationships between the expression of these markers and patient evolution following conventional therapy was investigated. The series consisted of paraffin-embedded samples from the Laboratory of Anatomical Pathology and Cytology of Amaral Carvalho Hospital in Jaú, SP, Brazil, of 103 patients diagnosed with testicular non-seminoma germ cell tumor (NSGCT) over a 15-year period (1996-2010). Among the 103 cases, testicular NSGCT predominated (57.3%), and among these, staining for EGFR and c-Kit predominated. EGFR was expressed when the choriocarcinoma component was predominant, and c-K... (Complete abstract click electronic access below) / Doutor

Despite the general advice of avoiding medication during pregnancy, the majority of pregnant woman use one or more ‘over the counter’ analgesics. During the last few years there has been growing evidence that analgesic exposure, such as paracetamol, ibuprofen or indomethacin, during pregnancy can have detrimental effects on rodent and human fetal gonads. The majority of previous studies have focused in alterations in testosterone production and male reproductive disorders. However, few studies have analysed the effect of these analgesics on fetal germ cells and possible consequences on fertility. During my thesis, I first focused on the effect of paracetamol and indomethacin exposure during pregnancy on rat fetal gonads. These showed that both paracetamol and indomethacin are able to alter the expression of genes important for fetal gonad and germ cell development. Previous studies on germ cells and analgesics have focused on rat models, but there is a lack of similar studies performed in human models. Therefore, I investigated the consequences of exposure of therapeutically relevant doses of paracetamol and ibuprofen on human gonads, with a special attention to the germ cells. Fetal gonads from the 1st and 2nd trimester were used in two different models: hanging drop cultures for 1st trimester testes and ovaries and a xenograft system for 2nd trimester fetal testes. Fetal gonad culture in the presence of paracetamol or ibuprofen reduced AP2γ+ (gonocyte) GC number in both 1st trimester fetal testes (22-28% reduction) and ovaries (43-49% reduction). 2nd trimester fetal testes were exposed to three different regimes, 1 or 7 days paracetamol and 7 days ibuprofen, which led to reductions of 17% and 30%, respectively in AP2γ+ GC number for paracetamol and a 53% reduction in total germ cell number for ibuprofen.

Made available in DSpace on 2014-06-11T19:24:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-28Bitstream added on 2014-06-13T19:52:03Z : No. of bitstreams: 1 rodrigues_am_me_botfm.pdf: 593199 bytes, checksum: 3862ac32735e6b1acf2a9269bf6223f8 (MD5) / Universidade Estadual Paulista (UNESP) / Com a finalidade de avaliar a repercussão no prognóstico e na morbidade dos pacientes com TCG em função da adesão do cirurgião ao protocolo cirúrgico, foram analisados retrospectivamente, com ênfase nos aspectos cirúrgicos, 64 pacientes de 0 a 18 anos completos, durante o período de junho de 1996 a janeiro de 2009 em dois centros de referência para o tratamento do paciente oncológico pediátrico: Hospital Amaral Carvalho de Jaú e o Hospital das Clínicas da Faculdade de Medicina de Botucatu (UNESP). Os dados coletados eram referentes a variáveis qualitativas e, desse modo, foram obtidas frequências e porcentagens para cada variável. As associações entre as variáveis de interesse foram feitas obtendo-se tabelas de contingência e testes qui-quadrado ou exato de Fisher. Em todos os testes foi adotado o nível de significância de 5%. Dos 64 pacientes estudados, 22 do Hospital Amaral Carvalho e 42 do Hospital das Clínicas – UNESP, com relação ao local primário, houve predomínio do ovário, seguido de testículo, sacrococcígeo, retroperitoneal, mediastino, orofaringe e útero. O tipo histológico predominante foi o teratoma puro, e o estadio I foi o mais freqüente. Quarenta e quatro pacientes foram classificados como baixo risco, e todos os 64 pacientes receberam tratamento cirúrgico, sendo em 44 deles exclusivo. A cirurgia pediátrica foi quem mais encaminhou casos operados dos serviços de origem, enquanto a ginecologia foi a especialidade que mais operou nas instituições de estudo. Quinze casos foram encaminhados com algum tipo de cirurgia e 49 encaminhados sem cirurgia. A quantidade de casos encaminhados sem cirurgia prévia foi estatisticamente significante quando comparado aos casos encaminhados com algum tipo de cirurgia. Dos 15 pacientes encaminhados com algum tipo de cirurgia, 10 seguiram o protocolo... / In order to assess the impact on prognosis and morbidity of patients with GCT according to the surgeon's accession to the surgical protocol were analyzed retrospectively, with emphasis on surgery, 64 patients 0-18 years of age, during the period of June 1996 to January 2009 in two hospitals for the treatment of pediatric cancer patients: Hospital Amaral Carvalho de Jau and the Hospital of the Medical School of Botucatu (UNESP). Data were collected regarding the qualitative variables, and thus, frequencies and percentages were obtained for each variable. The associations between the variables of interest were made by obtaining contingency tables and chisquare or Fisher exact test. In all tests, we adopted the significance level of 5%. Of the 64 patients studied, 22 of 42 Amaral Carvalho Hospital and the Hospital das Clinicas - UNESP, with respect to the primary site, there was a predominance of the ovary, followed by testis, sacrococcygeal, retroperitoneal, mediastinal, oropharynx and uterus. The predominant histological type was pure teratoma, and the stage I was the most frequent. Forty-four patients were classified as low risk, and all 64 patients received surgical treatment in 44 of them being unique. The pediatric surgery was the most referred cases operated source of services, while gynecology was the specialty that most institutions operated in the study. Fifteen cases were referred to some type of surgery and 49 conveyed without surgery. The number of cases referred without previous surgery was statistically significant when compared to cases presenting with some type of surgery. Of the 15 patients referred with type of surgery, 10 followed the protocol and not 5, while the 49 patients referred without surgery, 24 were operated on following the protocol and not 25. Although there was no statistically significant differences... (Complete abstract click electronic access below)

Submitted by FRANCISCO CARLOS QUEVEDO null (fcqjau@gmail.com) on 2017-09-15T16:19:40Z No. of bitstreams: 1 _TTCG FINAL 260715 FCQ [2].pdf: 2024513 bytes, checksum: 9e2b5af27bf3e7a02eb1625af9c3c0fc (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-09-15T16:27:51Z (GMT) No. of bitstreams: 1 quevedo_fc_dr_bot.pdf: 2024513 bytes, checksum: 9e2b5af27bf3e7a02eb1625af9c3c0fc (MD5) / Made available in DSpace on 2017-09-15T16:27:51Z (GMT). No. of bitstreams: 1 quevedo_fc_dr_bot.pdf: 2024513 bytes, checksum: 9e2b5af27bf3e7a02eb1625af9c3c0fc (MD5) Previous issue date: 2017-08-25 / As neoplasias testiculares constituem aproximadamente 1% dos cânceres masculinos. Dentre eles, os tumores que têm origem nas células germinativas (TTCG) são os mais frequentes. Por sua natureza, esses tumores têm padrão morfológico variado, sendo distribuídos em dois subgrupos: os seminomas e os não-seminomas. Esses últimos, por sua vez, são classificados em carcinomas embrionários, tumores do seio endodérmico, coriocarcinomas, teratomas e tumores mistos de células germinativas. Há consenso na literatura de que a invasão vascular verificada histologicamente no tumor primário é o melhor indicador preditivo de progressão da doença e recidiva. Estudos recentes demonstram o papel dos fatores de crescimento e seus receptores para avaliação prognóstica dos TTCG, no que o uso da imuno-histoquímica é fundamental. O presente estudo procurou avaliar pela técnica imuno-histoquímica à ocorrência da expressão do receptor do fator de crescimento epidérmico (EGFR), do Her2 e do c-Kit em uma série de TTCG não-seminomas primários de testículo. Além disso, explorou a possível relação entre a expressão desses marcadores com a evolução dos pacientes após terapêutica convencional. A série foi constituída de amostras parafinadas existentes no Laboratório de Anatomia Patológica e Citologia do Hospital Amaral Carvalho de Jaú, totalizando 103 pacientes que receberam o diagnóstico de tumor testicular de células germinativas não-seminomas (TTCGNS) no período 1996-2010. Dentre os 103 casos, predominaram os TTCGMNS (57,3%). Nestes, observou-se o predomínio de marcação para EGFR e c-Kit. O EGFR foi expresso quando o componente coriocarcinoma era predominante e o c-Kit no componente epitelial do teratoma. A hipótese do estudo, que os casos positivos para os marcadores analisados estariam correlacionados com sobrevida menor, não foi confirmada pela analise estatística de Kaplan-Meier; seria necessário um maior número de casos para se chegar a uma conclusão quanto à sobrevida. Superexpressão, amplificação gênica e mutações ativadoras são frequentes em tumores testiculares germinativos; deste modo, o uso da técnica de Hibridização Fluorescente in situ (FISH) foi utilizada para avaliação dos eventos gênicos relativos a imunoexpressão proteica, observada pelos resultados encontrados no estudo imuno-histoquímico. Somente um caso, classificado como TTCGM, apresentava amplificação para gene EGFR. Este caso apresentou bom prognóstico. Considerou-se que, a ausência de amplificação observada na maioria dos casos, deve-se a qualidade inadequada do material parafinado e estocado. Conclui-se que, na série estudada, os TTCGNS expressam os marcadores EGFR, Her2 e c-Kit, a depender do subtipo histológico; entretanto, estas proteínas não tiveram impacto prognostico. A amplificação gênica para o EGFR pode ocorrer de forma isolada, porem devido a condições técnicas restritivas, não foi possível observa-la nos demais casos positivos para EGFR pela técnica de imuno-histoquímico. / Testicular neoplasms constitute approximately 1% of male cancers. Among these, germ cell tumors (TGCT) are the most prevalent. These tumors show varied morphological patterns and are distributed into two subgroups: seminomas and non-seminomas. The latter, in turn, are classified as embryonal carcinomas, endodermal sinus tumors, choriocarcinomas, teratomas and mixed germ cell tumors. The consensus in the literature is that histological verification of vascular invasion (VI) in the primary tumor is the best predictor of disease progression and recurrence. However, recent studies have highlighted the role of growth factors and their receptors for prognostic evaluation of TGCT, in which the use of immunohistochemistry is fundamental. This study sought to evaluate the occurrence of epidermal growth factor receptor (EGFR), Her2 and c-Kit in a series of primary non-seminoma TGCTs. In addition, possible relationships between the expression of these markers and patient evolution following conventional therapy was investigated. The series consisted of paraffin-embedded samples from the Laboratory of Anatomical Pathology and Cytology of Amaral Carvalho Hospital in Jaú, SP, Brazil, of 103 patients diagnosed with testicular non-seminoma germ cell tumor (NSGCT) over a 15-year period (1996-2010). Among the 103 cases, testicular NSGCT predominated (57.3%), and among these, staining for EGFR and c-Kit predominated. EGFR was expressed when the choriocarcinoma component was predominant, and c-Kit when the epithelial component of the teratoma was predominant. The initial hypothesis that positive cases for any marker would be correlated to lower survival was not confirmed by Kaplan-Meier survival probability studies. A larger number of cases is required to reach any conclusions concerning survival. Gene overexpression and amplification and activating mutations are common in testicular germinal tumors, thus fluorescent in situ hybridization (FISH) was used to validate the genic events results obtained in the immunohistochemical study. Only one case, classified as mixed TGCT, showed EGFR gene amplification, and this case presented good prognosis. The lack of amplification observed in most cases was due to the inadequate quality of the stored, paraffinembedded materials. In conclusion, in the series studied, testicular NSGCT expressed EGFR, Her2 and c-Kit markers depending on the histological subtype; however, these proteins had no prognostic impact. Gene amplification for EGFR may occur in isolation, but due to the limited technical conditions, it was not observed in the other cases positive for EGFR using immunohistochemistry.

Orientador: Érika Veruska Paiva Ortolan / Banca: Bonifácio Katsumori Takegawa / Banca: José Roberto de Souza Baratella / Resumo: Com a finalidade de avaliar a repercussão no prognóstico e na morbidade dos pacientes com TCG em função da adesão do cirurgião ao protocolo cirúrgico, foram analisados retrospectivamente, com ênfase nos aspectos cirúrgicos, 64 pacientes de 0 a 18 anos completos, durante o período de junho de 1996 a janeiro de 2009 em dois centros de referência para o tratamento do paciente oncológico pediátrico: Hospital Amaral Carvalho de Jaú e o Hospital das Clínicas da Faculdade de Medicina de Botucatu (UNESP). Os dados coletados eram referentes a variáveis qualitativas e, desse modo, foram obtidas frequências e porcentagens para cada variável. As associações entre as variáveis de interesse foram feitas obtendo-se tabelas de contingência e testes qui-quadrado ou exato de Fisher. Em todos os testes foi adotado o nível de significância de 5%. Dos 64 pacientes estudados, 22 do Hospital Amaral Carvalho e 42 do Hospital das Clínicas - UNESP, com relação ao local primário, houve predomínio do ovário, seguido de testículo, sacrococcígeo, retroperitoneal, mediastino, orofaringe e útero. O tipo histológico predominante foi o teratoma puro, e o estadio I foi o mais freqüente. Quarenta e quatro pacientes foram classificados como baixo risco, e todos os 64 pacientes receberam tratamento cirúrgico, sendo em 44 deles exclusivo. A cirurgia pediátrica foi quem mais encaminhou casos operados dos serviços de origem, enquanto a ginecologia foi a especialidade que mais operou nas instituições de estudo. Quinze casos foram encaminhados com algum tipo de cirurgia e 49 encaminhados sem cirurgia. A quantidade de casos encaminhados sem cirurgia prévia foi estatisticamente significante quando comparado aos casos encaminhados com algum tipo de cirurgia. Dos 15 pacientes encaminhados com algum tipo de cirurgia, 10 seguiram o protocolo... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In order to assess the impact on prognosis and morbidity of patients with GCT according to the surgeon's accession to the surgical protocol were analyzed retrospectively, with emphasis on surgery, 64 patients 0-18 years of age, during the period of June 1996 to January 2009 in two hospitals for the treatment of pediatric cancer patients: Hospital Amaral Carvalho de Jau and the Hospital of the Medical School of Botucatu (UNESP). Data were collected regarding the qualitative variables, and thus, frequencies and percentages were obtained for each variable. The associations between the variables of interest were made by obtaining contingency tables and chisquare or Fisher exact test. In all tests, we adopted the significance level of 5%. Of the 64 patients studied, 22 of 42 Amaral Carvalho Hospital and the Hospital das Clinicas - UNESP, with respect to the primary site, there was a predominance of the ovary, followed by testis, sacrococcygeal, retroperitoneal, mediastinal, oropharynx and uterus. The predominant histological type was pure teratoma, and the stage I was the most frequent. Forty-four patients were classified as low risk, and all 64 patients received surgical treatment in 44 of them being unique. The pediatric surgery was the most referred cases operated source of services, while gynecology was the specialty that most institutions operated in the study. Fifteen cases were referred to some type of surgery and 49 conveyed without surgery. The number of cases referred without previous surgery was statistically significant when compared to cases presenting with some type of surgery. Of the 15 patients referred with type of surgery, 10 followed the protocol and not 5, while the 49 patients referred without surgery, 24 were operated on following the protocol and not 25. Although there was no statistically significant differences... (Complete abstract click electronic access below) / Mestre

With meiosis spanning from embryonic development to the end of reproductive life in females, scientists have faced considerable limitations in studying female meiosis and the effects of toxicants on the developing oocyte. Over the last half century, various culture methods have been developed with the aim of studying the mechanisms of early ovary development, as well as for use in reproductive toxicology. However, very few of the established embryonic ovary culture systems have been used to investigate potential reproductive toxicants on the embryonic ovary, in particular when compared with the vast number of in vitro reproductive toxicity studies on the post-natal ovary. Here, a novel test compound, a topoisomerase II inhibitor: AstraZeneca Test Compound (AZTC), was used to investigate the efficacy and validity of ovarian culture methods when compared with in vivo reprotoxicity studies. AZTC was selected due to preliminary in vivo studies demonstrating its detrimental effects on spermatogenesis in male rats. AZTC targets bacterial type II topoisomerases that might have mammalian homologues involved in meiosis. Topoisomerase-II α was expressed within the female germ cells pre-natally, but became localised to the granulosa and stroma cells post-natally. This occurred both in vivo and in vitro. Ovaries from female rats exposed pre-natally to AZTC in vivo were analysed histologically and a significant increase in the number of primordial follicles was observed within the ovaries, as well as an increase in the number of unhealthy follicles. A novel mouse embryonic ovary culture system was developed by adapting, improving and bridging existing available culture techniques. The culture system supported growth of pre-meiotic mouse germ cells through prophase I of meiosis, the formation of primordial follicles and initiation of follicle growth. Cultured ovaries contained follicles at stages in comparable ratios to those in vivo and appeared morphologically normal and healthy. The culture also supported meiotic progression of oocytes to the pachytene stage, albeit with a slight delay. AZTC was used to validate the novel embryonic ovary culture by comparing the results with those from the in vivo study, where AZTC exposure had also occurred during embryonic development. Similar results were consistently observed between the in vivo and in vitro studies. In vitro effects of AZTC on the post-natal mouse ovary were also investigated, where neonatal mouse ovaries cultured with AZTC had fewer primordial follicles and more unhealthy follicles than did control ovaries. AZTC therefore demonstrated different effects when exposure occurred pre-natally vs. post-natally. The embryonic ovary culture was then used to examine the effects of another topoisomerase II inhibitor, etoposide, on the pre-natal ovary. Etoposide is a chemotherapy agent and has previously been prescribed to pregnant women. A significant reduction in the size of the follicle pool was observed in exposed cultured embryonic ovaries, where primordial and transitional follicles were targeted. Overall, establishment of post-natal culture systems have become a useful addition to in vivo reproductive toxicology studies. The embryonic ovary culture system developed here could become a valuable and powerful tool to screen potential reproductive toxicants, as well as to study the dynamics and regulation of early ovary development.