Studies of prophylactic salpingectomy specimens from BRCA1/2 mutation carriers, at risk for tubal and ovarian high-grade serous carcinoma (SerCa), have consistently revealed occult carcinomas and putative histological cancer precursors in the distal fallopian tube epithelium (FTE), supporting the FTE as the source of SerCa. In this thesis I molecularly characterized and compared non-malignant FTE from mutation carriers (FTEb) and control patients (FTEn) to identify alterations that may predispose to malignant transformation. Gene expression profiling of laser capture microdissected FTEn, FTEb and SerCa indicated that SerCa have similar molecular profiles whether of presumed ovarian or tubal origin, supporting the notion they share a common cell of origin within the FTE. Furthermore, FTEb samples obtained during the post-ovulatory luteal phase showed gene expression profiles closely resembling SerCa samples, suggesting that the luteal phase milieu may contribute to serous carcinogenesis. An initial hypothesis was that FTEb may respond differently to luteal progesterone compared to FTEn, via differential expression of progesterone receptor (PR) isoforms. However, similar relative isoform expression in FTEn and FTEb samples suggested that a luteal phase-associated factor other than progesterone directs gene expression changes in FTEb. The possibility that FTEb respond differently to ovulation-associated inflammatory cytokines that are locally elevated during the luteal phase was next investigated. Importantly, FTEb specimens previously found to cluster with SerCa based on their global gene expression profiles showed evidence of increased nuclear factor-κB (NFκB)-dependent (pro-inflammatory) signalling and diminished glucocorticoid receptor (GR)-dependent (anti-inflammatory) signalling. Furthermore, I demonstrate that disabled homolog 2 (DAB2), an adaptor molecule decreased in SerCa and FTE luteal samples, enhances both GR-mediated transactivation and suppression of NFκB signalling, implicating DAB2 as a crucial determinant of inflammatory signalling and ovarian cancer risk. Altogether, this thesis identifies gene expression changes in FTE from BRCA mutation carriers during the post-ovulatory luteal phase that parallel those detected in SerCa. The data support a proposed novel testable model for predisposing events contributing to SerCa that centres on an altered ability to quickly resolve the pro-inflammatory environment created by the ovulatory event.

Studies of prophylactic salpingectomy specimens from BRCA1/2 mutation carriers, at risk for tubal and ovarian high-grade serous carcinoma (SerCa), have consistently revealed occult carcinomas and putative histological cancer precursors in the distal fallopian tube epithelium (FTE), supporting the FTE as the source of SerCa. In this thesis I molecularly characterized and compared non-malignant FTE from mutation carriers (FTEb) and control patients (FTEn) to identify alterations that may predispose to malignant transformation. Gene expression profiling of laser capture microdissected FTEn, FTEb and SerCa indicated that SerCa have similar molecular profiles whether of presumed ovarian or tubal origin, supporting the notion they share a common cell of origin within the FTE. Furthermore, FTEb samples obtained during the post-ovulatory luteal phase showed gene expression profiles closely resembling SerCa samples, suggesting that the luteal phase milieu may contribute to serous carcinogenesis. An initial hypothesis was that FTEb may respond differently to luteal progesterone compared to FTEn, via differential expression of progesterone receptor (PR) isoforms. However, similar relative isoform expression in FTEn and FTEb samples suggested that a luteal phase-associated factor other than progesterone directs gene expression changes in FTEb. The possibility that FTEb respond differently to ovulation-associated inflammatory cytokines that are locally elevated during the luteal phase was next investigated. Importantly, FTEb specimens previously found to cluster with SerCa based on their global gene expression profiles showed evidence of increased nuclear factor-κB (NFκB)-dependent (pro-inflammatory) signalling and diminished glucocorticoid receptor (GR)-dependent (anti-inflammatory) signalling. Furthermore, I demonstrate that disabled homolog 2 (DAB2), an adaptor molecule decreased in SerCa and FTE luteal samples, enhances both GR-mediated transactivation and suppression of NFκB signalling, implicating DAB2 as a crucial determinant of inflammatory signalling and ovarian cancer risk. Altogether, this thesis identifies gene expression changes in FTE from BRCA mutation carriers during the post-ovulatory luteal phase that parallel those detected in SerCa. The data support a proposed novel testable model for predisposing events contributing to SerCa that centres on an altered ability to quickly resolve the pro-inflammatory environment created by the ovulatory event.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 5 uppsatser.

Orientador: Luis Guillermo Bahamondes / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-19T05:07:00Z (GMT). No. of bitstreams: 1 Hermanny_Alexia_D.pdf: 843759 bytes, checksum: 4b2ff42ff4ef1f995e85e5f8b86bf7f2 (MD5) Previous issue date: 2011 / Resumo: O mecanismo de ação do levonorgestrel (LNG) na anticoncepção de emergência (AE) ainda não está totalmente esclarecido e seu efeito nas funções espermáticas também não está explicado. Os objetivos deste trabalho foram avaliar se o LNG, em dose igual à observada após a ingestão oral para AE, poderia afetar espermatozoides expostos in vitro à tuba uterina humana e realizar uma revisão bibliográfica sobre o efeito do LNG nas diferentes funções espermáticas. Foram realizados 15 experimentos. As tubas uterinas foram removidas através de mini-laparotomias e foram perfundidas com uma suspensão contendo 1x106 espermatozoides móveis, com e sem LNG. A tuba correspondente ao lado onde o folículo dominante estava presente recebeu a suspensão com LNG em pacientes alternados. Após um período de incubação de 4 horas, o istmo e a ampola de cada tuba foram separados. Cada segmento foi lavado separadamente e o material obtido foi avaliado quanto ao número de espermatozoides móveis recuperados, número de espermatozoides aderidos ao epitélio tubário e taxa de reação acrossômica (RA). A presença do LNG não afetou significativamente o número de espermatozoides móveis recuperados do istmo e da ampola, e não afetou o número de espermatozoides aderidos ao epitélio tubário. O LNG também não influenciou a taxa de RA. Diferenças significativas também não foram observadas quando o lado ovulatório foi considerado. A revisão bibliográfica realizada deixou evidente que existem poucos estudos que analisam a influência do LNG como AE sobre as funções espermáticas, apesar de este ser um possível mecanismo de ação. Além disso, os estudos revisados utilizam diferentes métodos de avaliação e os resultados são, muitas vezes, contraditórios. De acordo com os resultados observados na literatura, quando o LNG é usado na AE, provavelmente não atinge concentração plasmática suficiente para ser reconhecido pelos receptores de progesterona (P). Resultados positivos só foram observados quando a dose de LNG utilizada nos experimentos foi comparável ao sistema intrauterino liberador de LNG (SIU-LNG), ou seja, muito maior que a utilizada na AE. O LNG, em dose similar à observada no plasma após a ingestão oral para AE, não afetou o número, a aderência ao epitélio tubário, a distribuição e a taxa de RA de espermatozoides na tuba uterina humana, in vitro. De acordo com os resultados observados na literatura, se o LNG, na concentração utilizada para AE, afeta ou não a função espermática ainda não está claro, e mais estudos são necessários / Abstract: The mechanism of action of levonorgestrel (LNG) as emergency contraception (EC) is still under debate and the effect upon sperm function is partially explained. The aim of this study was to assess if LNG in a similar dose to those observed in serum after oral intake for EC could affect the spermatozoa when exposed in vitro to human tubes and also to give an overview of the effect of LNG as EC on several sperm functions. Fifteen mini-laparotomies were performed, the ovulatory side was recorded and both tubes were removed and perfused with a suspension of 1x106 of motile spermatozoa, one with LNG and the other without it. After an incubation period of 4hours the tubes were cut to separate the isthmus and the ampulla. Each segment was flushed and the material was evaluated regarding the motile sperm number, the number of spermatozoa adhering to the oviductal epithelium and acrosome reaction (AR) rate. The addition of LNG did not significantly affect the number of recovered spermatozoa neither at the isthmus nor at the ampulla or the number of recovered spermatozoa adhered at the human tubal epithelium. Additionally, LNG did not influence the rate of AR. There were no significant differences even when the ovulatory side was taken into account. The present review showed that there are few studies which focus on the influence of LNG as EC upon sperm functions; albeit it is a plausible mechanism of action. Additionally, the different studies used different methods of evaluation and the results were in many cases contradictories. According to the results observed at the literature, when LNG is used as EC, it is probable that the drug does not achieved sufficient serum concentrations in order to be recognized by the progesterone (P) receptors. Positive results only were observed when the dose of LNG used in the experiments was much higher (comparable to the LNG-IUS) than the proposed for EC. LNG in a similar dose to that observed in serum after oral intake for EC did not affect the number, the adhesion to tubal epithelium, distribution, and AR rate of spermatozoa at the human Fallopian tubes in vitro. According to the results observed at the literature, if the LNG in doses used for EC, affects sperm function or not, it is still uncertain and warrants further studies / Doutorado / Fisiopatologia Ginecológica / Doutor em Ciências da Saúde

OBJECTIVE: To determine if laser-assisted anastomosis is superior to microsurgical anastomosis and laser welding for tubal reconstruction. STUDY DESIGN: Sixty uterine tubes from 30 rabbits were transected and then anastomosed with a microsurgical technique, laser welding and laser-assisted anastomosis using a microscope. The rabbits were mated one month postoperatively and examined for implantation sites and adhesion formation three to seven days postpartum. RESULTS: The amount of time required to perform laser-assisted anastomosis as well as laser welding was significantly shorter than for microsurgery. All the tubes withstood the distension pressure of pregnancy with the exception of 1 of the 20 laser-welded tubes, which had 30% of its circumference dehisced. There was no difference in the number of implantation sites per tube between the different groups. The anastomotic sites were well healed and were nonidentifiable except for holding stitches and microsurgical sutures. CONCLUSION: Laser-assisted anastomosis and laser welding took less time to perform than microsurgery. Laser-assisted anastomosis resulted in excellent healing, as did microsurgery. With the protection of serum albumin, laser-assisted anastomosis did not cause any thermal damage, and the anastomotic sites could tolerate the distension pressure of pregnancy and parturition without problems. Laser welding without protection of serum albumin could cause thermal damage and dehiscence. The implantation and pregnancy rates were comparable The implantation and pregnancy rates were comparable with all three types of procedure.

As Distrofias Musculares Progressivas (DMPs) constituem um grupo de doenças genéticas caracterizadas por uma degeneração progressiva e irreversível da musculatura esquelética. Dentre as diferentes abordagens terapêuticas propostas para esse grupo de doenças, a terapia celular com células-tronco mesenquimais (CTMs) tem sido um foco importante de pesquisas. Muitos tipos de CTMs já foram descritas com o intuito de reconhecer qual o tipo ideal a ser usado em uma possível terapia celular para DMPs. Neste trabalho comparamos o potencial terapêutico de células-tronco de diferentes fontes obtidas de um mesmo doador. Escolhemos as células de pericito (CP) como ferramenta de estudo, uma vez que elas estão presentes em todos os tecidos irrigados por vasculatura. Isolamos pericitos de 4 tecidos da mesma doadora (endométrio, trompa, tecido adiposo e músculo). Em seguida, injetamos 1 milhão dessas células intraperitonialmente em camundongos Utrntm1KedDmdmdx/J (duplo knockout para o gene da distrofina e utrofina) semanalmente, por 8 semanas, e avaliamos a clínica e a sobrevida desses animais. Observamos que nas condições experimentais desse estudo, o potencial miogênico dessas células é insuficiente para ser utilizado como terapia regenerativa. Entretanto, apesar dos testes padronizados não detectarem nenhuma melhora clínica aparente, os animais tratado com pericitos de gordura mostraram uma curva de sobrevivência significativamente maior do que os controles não tratados. Como não houve diferenciação miogênica, esses resultados sugerem que os efeitos benéficos ocorreram através de liberação de fatores tróficos e imunoreguladores (efeito parácrino). É digno de nota que apesar das células serem todas derivadas de pericito e de uma mesma doadora o aumento de sobrevida só foi observado com células do tecido adiposo. Esses resultados indicam que o potencial terapêutico de CP difere de acordo com sua origem e que essa diferença não depende do genoma do doador. Esses resultados constituem um passo inicial, porém, valioso na compreensão do potencial de utilização dessas células em terapias / Progressive Muscular Disorders (PMDs) are a group of heterogeneous genetic diseases characterized by an irreversible degeneration of the muscle tissue due to mutation or absence of a protein. Among the many different available therapeutic approaches to treat PMDs, cell therapy using mesenchymal stem cells (MSCs) are one of the most studied ones. There are many types of MCSs described to date and the need to identify the best one suited to treat PMDs has yet to be addressed. In this thesis, we compared the therapeutic potential of different types of stem cells derived from the same donor. First, we chose pericytes as a tool of comparison, as this cell is unequivocally present in all vascularized tissues. We isolated pericytes of 4 different tissues from the same donor (endometrium, fallopian tubes, adipose tissue and muscle). We injected 1 million of these cells intraperitonially in Utrntm1KedDmdmdx/J mice (knockout for the dystrophin and utrophin gene) weekly for 8 weeks evaluating the clinical features and survival curve of these mice. We observed that, in the experimental conditions of this study, the myogenic potential of these cells is insufficient to be harnessed as therapy for regenerative purposes. However, despite the fact that the standardized tests did not detect any apparent clinical improvement, mice treated with pericytes derived from adipose tissue had a survival curve greater than control treated mice. As we could not observe any myogenic differentiation or cell engraftment, this results suggests that the beneficial effect observed could be due to the releasing of trophic and immune modulator factors (paracrine effect). It is noteworthy that despite all cells being derived from the same donor, the increase in life expectancy was only observed in pericytes derived from the adipose tissue. These results indicate that the therapeutic potential of pericytes differs according to their tissue of origin and the difference is not due to genetic differences. This is still preliminary data but it is valuable in understanding the therapeutic potential of these cells

Zellkultursysteme mit Krebszelllinien werden seit Langem zur Untersuchung der Interakti-on zwischen Pathogenen und ihren Wirtszellen eingesetzt. Diese Systeme eignen sich aufgrund der reduzierten Komplexität für die Analyse einzelner Faktoren, spiegeln jedoch nicht den Zustand primärer Zellen oder die komplexe Gewebestruktur wieder. Um die Beschränkungen zu umgehen, wurden in dieser Arbeit neue Modelle etabliert auf der Grundlage von reversibel immortalisierten humanen Primärzellen und ex vivo Kultur von intaktem humanem Eileitergewebe. Infektionen mit dem humanpathogenen Bakterium Chlamydia trachomatis, welches chronische Schmerzen oder Unfruchtbarkeit auslösen kann, wurden in diesen Modellen untersucht. Reversible Immortalisierung wurde mit pri-mären human Eileiterzellen (FT Zellen) und humanen Nabelschnurzellen (HUVEC) durchgeführt. Das System basiert auf lentiviralem Gentransfer und dem Cre-lox-System. HUVEC Zellen wurden mit Kombinationen der Onkoproteine hTERT, SV40T und Bmi1 immortalisiert. Immortalisierung von FT Zellen wurde mit SV40T und Bmi1 erreicht. Eine Analyse der FT Zelllinien zeigte Veränderungen des Karyotyps durch die Immortalisie-rung. Bemerkenswerterweise konnten die Stammzellmarker CD44 und Oct4 in FT Zellen nachgewiesen werden. Ex vivo Gewebekultur humaner Eileiter wurde als stabiles Infekti-onsmodel für Chlamydia trachomatis etabliert. Mittels hochauflösender Konfokal-mikroskopie wurde gezeigt, dass die Infektion mit C. trachomatis tiefgreifende Verände-rungen im Epithel der Mukosa auslöst und zum Verlust der Zelladhäsion und Zellpolarität führt. Ein erhöhter Anteil apoptotischer Zellen wurde nach Infektion mit Serovar D beo-bachtet, einem klinischen Isolat des Genitaltraktes. Dieses Ergebnis steht im Gegensatz zu Infektionen mit dem Laborstamm Serovar L2. Phänotypische Veränderungen in nicht infizierten Zellen weisen auf die Existenz parakriner Signalwege während der akuten In-fektion und Veränderung der epithelialen Homeostase hin. / Cell culture systems with cancer-derived cell lines have long been used to study the interaction between pathogens and their host cells. Due to reduced complexity these systems are convenient for the analysis of single factors; however, they do not represent the condition of primary cells or the complex tissue structure. To circumvent these limitations new models were established in this study on the basis of reversibly immortalized human primary cells and ex vivo culture of intact human fallopian tube tissue. Infections with the human pathogenic bacterium Chlamydia trachomatis, which can lead to chronic pain or infertility, were analyzed in these models. Reversible immortalization was applied to primary human fallopian tube (FT) cells and human umbilical vein cells (HUVEC). This system is based on lentiviral gene transfer and the Cre-lox-system. HUVEC cells were immortalized with a combination of two of the oncoproteins hTERT, SV40T and Bmi1. Immortalization of FT cells was achieved with SV40T and Bmi1. Analysis of FT cell lines revealed changes of the karyotype induced by immortalization. Remarkably, the stem cell markers CD44 and Oct4 were detected in FT cells. Ex vivo tissue culture of human fallopian tubes was established as stable and reliable infection model for Chlamydia trachomatis. Via high resolution confocal analysis the infection with C. trachomatis was discovered to trigger profound changes in the epithelial mucosa, causing loss of cell adhesion and polarity. Interestingly, an increase in the rate of apoptotic cells was observed after infection with serovar D, a clinical genital tract isolate. This finding is in contrast to infections with serovar L2, a laboratory strain. Phenotypic changes in non-infected cells suggest the existence of paracrine signalling during acute infection and change in epithelial homeostasis.

Objetivo: Avaliar a correlação entre a concentração sérica e a expressão tecidual do fator de crescimento endotelial vascular (VEGF) na interface materno embrionária e avaliar a associação entre a profundidade da invasão trofoblástica e a expressão tecidual do VEGF em gestações ectópicas ampulares. Métodos: Estudo prospectivo com 34 mulheres com o diagnóstico de gestação ectópica ampular espontaneamente concebidas, submetidas à salpingectomia no período de 11 de julho de 2012 a 19 de agosto de 2013. Os critérios de exclusão foram: diagnóstico de gestação tubária não ampular, impossibilidade de coleta de sangue para a dosagem do VEGF sérico, da análise anatomopatológica ou da dosagem do VEGF na peça cirúrgica. Após a confirmação diagnóstica de gestação tubária e antes da realização da salpingectomia foram dosadas as concentrações séricas maternas de VEGF pela técnica de luminex. Após o procedimento cirúrgico foi analisada a expressão tecidual de VEGF por imuno-histoquímica e pela técnica de point-counting. Histologicamente, a invasão trofoblástica na parede tubária foi classificada em grau I: limitada à mucosa da tuba uterina; grau II: até a camada muscular; grau III: invasão de toda a espessura da tuba uterina. Resultados: 8 pacientes apresentaram invasão grau I, 7 pacientes com grau II e 19 com invasão grau III. Não se observou diferença estatisticamente significativa na associação entre o VEGF tecidual e os diferentes graus de invasão trofoblástica (p= 0,621) e a correlação do VEGF tecidual com a sua dosagem sérica pelo coeficiente de Spearman (Rho = -0,057) foi fraca (p= 0,748). Foi realizado modelo de regressão logística para comparar o desempenho do VEGF tecidual como fator de predição da profundidade da invasão do trofoblasto na parede tubária, comparando o grau I vs II e III e o grau III vs I e II e não houve diferença estatisticamente significativa. Conclusões: em gestações ectópicas ampulares, a profundidade da invasão do tecido trofoblástico na parede tubária não está associada com a concentração tecidual do VEGF e não há correlação entre as suas dosagens sérica e tecidual / Objetive: To evaluate the correlation between serum concentration and tissue expression of vascular endothelial growth factor (VEGF) at the feto-maternal interface and to associate the depth of trophoblastic invasion into the tubal wall with tissue VEGF in ampullary pregnancies. Methods: A prospective study was conducted on 34 spontaneously conceived ampullary ectopic pregnancies that were submitted to salpingectomy between July 11th, 2012 and August 19th, 2013. Exclusion criteria were: diagnosis of not ampullary pregnancy, impossibility of collecting blood sample for serum VEGF determination, impossibility of either anatomopathological or tissue VEGF analysis. Maternal serum VEGF concentrations were measured after the diagnosis confirmation and before salpingectomy. After surgery, tissue VEGF expression was measured by immunohistocemistry and point counting technique. Histologically, trophoblastic invasion into the tubal wall was classified as grade I when limited to the tubal mucosa, grade II when reaching the muscle layer and grade III when comprising the full thickness of the tubal wall. Results: 8 patients had tubal infiltration grade I, 7 had grade II and 19 had grade III infiltration. Depth of trophoblastic invasion was not associated with tissue VEGF expression (p= 0.621). The correlation between tissue expression and serum VEGF concentration was not present (Spearman\'s coefficient, Rho = -0.057) (p= 0.748). Logistic regression showed that tissue VEGF expression was not predictor of trophoblastic invasion and there was not statistically significant difference when comparing grade I vs II and III and grade III vs I and II. Conclusions: in ampullary pregnancies, the depth of trophoblastic penetration into the tubal wall is not associated with tissue VEGF expression. Serum VEGF do not show any correlation with tissue expression

Objetivo: Avaliar a correlação entre a concentração sérica e a expressão tecidual do fator de crescimento endotelial vascular (VEGF) na interface materno embrionária e avaliar a associação entre a profundidade da invasão trofoblástica e a expressão tecidual do VEGF em gestações ectópicas ampulares. Métodos: Estudo prospectivo com 34 mulheres com o diagnóstico de gestação ectópica ampular espontaneamente concebidas, submetidas à salpingectomia no período de 11 de julho de 2012 a 19 de agosto de 2013. Os critérios de exclusão foram: diagnóstico de gestação tubária não ampular, impossibilidade de coleta de sangue para a dosagem do VEGF sérico, da análise anatomopatológica ou da dosagem do VEGF na peça cirúrgica. Após a confirmação diagnóstica de gestação tubária e antes da realização da salpingectomia foram dosadas as concentrações séricas maternas de VEGF pela técnica de luminex. Após o procedimento cirúrgico foi analisada a expressão tecidual de VEGF por imuno-histoquímica e pela técnica de point-counting. Histologicamente, a invasão trofoblástica na parede tubária foi classificada em grau I: limitada à mucosa da tuba uterina; grau II: até a camada muscular; grau III: invasão de toda a espessura da tuba uterina. Resultados: 8 pacientes apresentaram invasão grau I, 7 pacientes com grau II e 19 com invasão grau III. Não se observou diferença estatisticamente significativa na associação entre o VEGF tecidual e os diferentes graus de invasão trofoblástica (p= 0,621) e a correlação do VEGF tecidual com a sua dosagem sérica pelo coeficiente de Spearman (Rho = -0,057) foi fraca (p= 0,748). Foi realizado modelo de regressão logística para comparar o desempenho do VEGF tecidual como fator de predição da profundidade da invasão do trofoblasto na parede tubária, comparando o grau I vs II e III e o grau III vs I e II e não houve diferença estatisticamente significativa. Conclusões: em gestações ectópicas ampulares, a profundidade da invasão do tecido trofoblástico na parede tubária não está associada com a concentração tecidual do VEGF e não há correlação entre as suas dosagens sérica e tecidual / Objetive: To evaluate the correlation between serum concentration and tissue expression of vascular endothelial growth factor (VEGF) at the feto-maternal interface and to associate the depth of trophoblastic invasion into the tubal wall with tissue VEGF in ampullary pregnancies. Methods: A prospective study was conducted on 34 spontaneously conceived ampullary ectopic pregnancies that were submitted to salpingectomy between July 11th, 2012 and August 19th, 2013. Exclusion criteria were: diagnosis of not ampullary pregnancy, impossibility of collecting blood sample for serum VEGF determination, impossibility of either anatomopathological or tissue VEGF analysis. Maternal serum VEGF concentrations were measured after the diagnosis confirmation and before salpingectomy. After surgery, tissue VEGF expression was measured by immunohistocemistry and point counting technique. Histologically, trophoblastic invasion into the tubal wall was classified as grade I when limited to the tubal mucosa, grade II when reaching the muscle layer and grade III when comprising the full thickness of the tubal wall. Results: 8 patients had tubal infiltration grade I, 7 had grade II and 19 had grade III infiltration. Depth of trophoblastic invasion was not associated with tissue VEGF expression (p= 0.621). The correlation between tissue expression and serum VEGF concentration was not present (Spearman\'s coefficient, Rho = -0.057) (p= 0.748). Logistic regression showed that tissue VEGF expression was not predictor of trophoblastic invasion and there was not statistically significant difference when comparing grade I vs II and III and grade III vs I and II. Conclusions: in ampullary pregnancies, the depth of trophoblastic penetration into the tubal wall is not associated with tissue VEGF expression. Serum VEGF do not show any correlation with tissue expression

Orientador: Francisco Eduardo Martinez / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-04T19:06:20Z (GMT). No. of bitstreams: 1 Almeida-Francia_CamilaContinDinizde_D.pdf: 36206900 bytes, checksum: d44823fb5bde1faf04aa09c33b7f7a87 (MD5) Previous issue date: 2005 / Resumo: O alcoolismo é o terceiro problema mais sério de Saúde Pública Mundial. A maioria é constituída de homens, entretanto, o índice de mulheres viciadas em bebidas alcoólicas vem aumentando. A tuba uterina promove o meio necessário para a maturação dos gametas femininos e masculinos (capacitação), a fertilização e o desenvolvimento inicial do embrião. As variedades UChA e UChB constituem modelos para os trabalhos relacionados aos aspectos genéticos, bioquímicos, fisiológicos, nutricionais e farmacológicos dos efeitos do álcool que são importantes fatores do alcoolismo humano. Dessa forma, o presente trabalho analisou o ciclo estral e as células epiteliais da tuba uterina da linhagem de ratos UChA e UChB, bebedores voluntários de etanol, divididos em três grupos: 1°) UChA de consumo voluntário de 6,05 glK.gIdia de etanol a 10%; 2°) UChB de consumo voluntário de 8,18 glK.gIdia de etanol a 100/Ó e 3j ratas Wistar (controle). Durante 20 dias seguidos procedeu-se à análise do ciclo estral através da coleta de esftegaços vaginais. Fêmeas em estro foram selecionadas para coleta e processamento dos órgãos, empregando-se técnicas microscópicas e morfométricas. Os grupos UChA e UChB apresentaram ciclo estral irregular. A região intramural (In) da tuba uterina dos ratos tratados apresentou vacúolos no citoplasma e cisternas dilatadas, núcleos com formatos irregulares e gotas lipídicas. A região do istmo (I) apresentou alteração de elétron-densidade dos microvilos e distribuição irregular dos cHios, além das características citadas na região In. A região da ampola (A) apresentou aumento de corpos lamelares e gotas lipídicas no citoplasma e na luz do epitélio. Da mesma forma, as regiões de infundíbulo (Inf) e fimbrias (F) dos ratos tratados apresentaram vacúolos e lacunas com conteúdo ou não em seu interior, mitocôndrias não preservadas, corpos lamelares, gotas lipídicas e cisternas dilatadas. Foram observados núcleos com formas irregulares e núcleos e conteúdo citoplasmático na luz do epitélio. A morfologia dos cHios apresentou-se alterada. O processo degenerativo adquiriu proporções crescentes do grupo UChA para o UChB. Os grupos tratados apresentaram menores áreas citoplasmáticas e nucleares comparados ao controle. Conclui-se que o etanol altera o ciclo estral e a estrutura das células epiteliais da tuba uterina. Devido às alterações, os processos de capacitação e fertilização estão comprometidos. Infere-se que devido à grande quantidade de lisosomos e formação de vacúolos, o álcool induza inicialmente morte celular diferente da apoptose / Abstract: Alcoholism is the third most serious problem of World Public Health. Although the majority consists of men, the index of women addicted to alcoholic beverages is increasing. The uterine tube promotes the necessary means for maturation of female and male gametes (capacitation), the fertilization and the initial development of the embI)'o. The varieties VChA and VChB constitute rare models for works related to genetic, biochemical, physiological, nutritional, and pharmacological effects of alcohol that are important factors in human alcoholism. Thus the present work analyzed the estrous cycle and the epithelial cells ofthe uterine tube (intramural- In, isthmus - I, ampulla - A, injUndibulum - Infandfimbrias - F) ofthe rat lineage VChA and VChB, voluntary drinkers of alcohol, divided into three groups: 1st) VChA, voluntary consumption of 6.05 gIKglday of ethanollO%; 2nd) VChB, voluntary consumption of8.18 gIKglday of ethanollO% and 3rd) Wistar rats (control). For 20 following days it was preceded to the ana1ysis ofthe estrous cycle through the collection of vaginal smears. Females in estrus were selected for collection and processing of organs, employing microscopic and morphometric techniques. The VChA and VChB rats show irregular estrous cycle. The In region of treated rats presented cytoplasmic vacuoles and dilated cisterna, nuclei with irregular forms and lipid droplets. The I region showed altered electrondensity and irregular distribution ofthe cilia, and the characteristics mentioned in the In region. The A region showed increase of lamellar bodies and lipid droplets in the cytoplasm and lumen of the epithelium. The Inf and F of treated rats showed vacuoles and lacunas with content or not in their interiors, non-preserved mitochondria, lamellar bodies, lipid droplets and dilated cisterna. There were observed nuclei with irregular forms and nuclei and cytoplasmic content in the lumen of the epithelium. Morphology of the cilia was observed to be altered. The degenera tive process acquired growing proportions ftom the VChA group to the VChB one. Treated groups presented smaller cytoplasmic and nuclear areas compared to the controI. It's concluded that ethanol alters the estrous cyc1e and the epithelial cell structure of uterine tube. Due the alterations, the fertilization and the capacitation process of ethanol-drinking rats are damaged. Yet it is inferred that due to the great quantity of lysosomes and formation of vacuoles, alcohol initially leads to cell death different ftom the apoptosis / Doutorado / Biologia Celular / Doutor em Biologia Celular e Estrutural