Orientador: Cassio Luis Zanettini Riccetto, Paulo Cesar Rodrigues Palma / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-09T14:36:06Z (GMT). No. of bitstreams: 1 Fraga_Rogeriode_D.pdf: 1486566 bytes, checksum: 87fb9bd342e742c5f6f95c1de1b15eea (MD5) Previous issue date: 2007 / Resumo: O período após a menopausa é associado com elevada incidência de sintomas no trato urinário baixo. Dentre os principais, destaca-se a síndrome da bexiga hiperativa e infecções urinárias recorrentes. Há uma dificuldade para separar os fatores relacionados à senescência daqueles associados com a menopausa, que podem interferir no aparecimento das disfunções miccionais na mulher idosa. A alteração mais significativa na menopausa é a deprivação estrogênica. Esta condição tem sugerido a indicação de estrogenioterapia no período pós-menopausa, com a finalidade de reduzir a sintomatologia urinária. Porém, as repercussões do hipoestrogenismo no trato urinário inferior ainda permanecem por ser estabelecidas. Este estudo foi conduzido utilizando-se 40 ratas Wistar (3 meses de idade) divididas em 4 grupos de 10 animais: Grupo 1: controle; Grupo 2: submetido à ooforectomia bilateral e após 4 semanas iniciada reposição sub-cutânea diária de 17 ß-estradiol durante 12 semanas; Grupo 3: procedimento SHAM e após 4 semanas iniciada reposição diária sub-cutânea de óleo de sésamo por 12 semanas e Grupo 4: submetidas à ooforectomia bilateral e sacrificadas após 12 semanas sem reposição hormonal. Foi utilizada técnica de imunohistoquímica para coloração das lâminas com anticorpo policlonal S-100. A aferição da densidade volumétrica das fibras nervosas na parede vesical foi efetuada através da estereologia utilizando-se o sistema ¿ teste G50. As ratas do Grupo IV, submetidas à ooforectomia sem reposição hormonal, apresentaram densidade volumétrica de fibras nervosas menor que os outros grupos (p<0.001 - teste de Newman-Keuls). Nos outros parâmetros analisados não houve diferença. A ooforectomia bilateral, em ratas, causou diminuição da densidade volumétrica de fibras nervosas vesicais. A administração de estradiol, no grupo castrado, apresentou diferença significativa nos parâmetros estereológicos analisados, em relação aos animais castrados sem reposição (p<0,001) / Abstract: The postmenopausal period is associated with a high incidence of symptoms in the lower urinary tract. The main urological bladder symptoms are urge incontinence and recurring urinary tract infections. It is hard to separate the influence of aging from that of menopause when studying the etiology of dysfunction in elderly women. The most significant result of menopause is estrogen deprivation. This fact has led to the use of hormonal replacement in postmenopausal women with lower urinary tract dysfunction. However, the best treatment in terms of type, dosage and method of estrogen administration has still not been defined. This study was conducted on 40 Wistar rats (3 months old). Group 1: remained intact; Group 2: underwent bilateral ovariectomy and after 4 weeks daily replacement of 17 ß-estradiol for 12 weeks; Group 3: sham operated and after 4 weeks daily replacement of sesame oil for 12 weeks and Group 4: underwent bilateral ovariectomy and were sacrified after 12 weeks. It was used imunohistochemistry evaluation using S-100 polyclonal antibody to stain the fibers on parafin rat bladder sections. The G50 stereologycal grid system was used to analyze the fibers. Ovariectomy had a decrease effect on the volumetric density of the neural fibers in the bladder wall. Estradiol replacement in castrated animals demonstrated a significative difference in the stereological parameters when compared to the castrated group without hormonal replacement. Long-term estrogen deprivation caused significant changes in bladder innervation, which can be characterized by a decreased number of nerves fibers by 65% (p<0.001) / Doutorado / Cirurgia / Doutor em Cirurgia

The ovarian hormone 17β-estradiol (E2) is one of the central regulators of the female reproductive system. E2 is also a pleiotropic regulator since it can exert its non-reproductive role on other organ systems. E2 is neuroprotective, it maintains body's energy homeostasis, participates in various repair mechanism and is required for neural development. However, there is a substantial evidence suggesting that there might be a molecular reprogramming of E2's action when it is supplied exogenously after E2 deprivation. Though the length of E2 deprivation and age has been linked to this phenomenon, the molecular components and how they activate this reprogramming is still elusive. Our main goal was to perform global proteomics and metabolomics study to identify the molecular components and their interaction networks that are being altered in the brain and serum after a short-term E2 treatment following ovariectomy (OVX) in Sprague Dawley rats. One of the strength of our global study is that it gave us extensive information on the brain proteome itself by identification of a wide number of proteins in different brain sections. By analyzing the differentially expressed proteins, our proteomics study revealed 49 different networks to be altered in 7 sections of the brain. Most of the perturbed networks were involved in cell metabolism, neural development, protein synthesis, cellular trafficking and degradation, and several stress response signaling pathways. We assessed the neuroenergetic status of the brain based on E2's response to various energy generating pathways, including glycolysis, TCA cycle, and oxidative phosphorylation, and several signaling pathways. All energetics pathways were shown to be downregulated in E2 treatment, which suggests that E2 exerts its neuroprotective role by restoring energy homeostasis in OVX rat model by regulating complex signaling and metabolic networks. Our second focus was to determine the metabolite response (amino acids and lipids) after E2 treatment in the brain and serum by employing targeted metabolomics study. We have found that in rat brain cortex there was significant upregulation of a wide number of amino acids suggesting alternate route of metabolism. Another alternate explanation is that E2 replacement replenished the amino acid pool in the tissue. Pathway enrichment analysis revealed upregulation of several pathways, including amino sugar metabolism, purine metabolism, and glutathione metabolism. By combining proteomics and metabolomics in two different biological matrices we were able to gather a vast array of information on how E2 replacement after E2 deprivation can confer neuroprotection. Our findings will help to create a foundation of basic science to be used for developing potentially effective hormone therapies.

published_or_final_version / Pharmacology and Pharmacy / Master / Master of Medical Sciences

Compararam-se as taxas de prenhez (TP) em vacas Nelore (Bos taurus indicus) utilizando implantes de norgestomet novos ou reutilizados associados ao valerato de estradiol (VE) e benzoato de estradiol (BE). Duzentos e quarenta e uma vacas Nelore amamentando foram separadas em dois grupos, e receberam implante auricular contendo 3 mg de norgestomet (n=122) ou implante de norgestomet (n=119) que já havia sido previamente utilizado por 10 dias. Os animais que receberam implante novo ou reutilizado, foram sub-divididos, e um lote recebeu uma injeção (IM) de 3 mg de norgestomet associados a 5 mg de valerato de estradiol e o outro, uma injeção (IM) de 50 mg de progesterona associados a 2 mg de benzoato de estradiol. No momento da retirada do implante todos os animais receberam 1 aplicação de análogo de PGF2&alpha; (IM) para assegurar a luteólise e 24 h após uma injeção de 1,0 mg de BE (IM). A inseminação artificial em tempo fixo (IATF) ocorreu 54-56 h após a retirada do implante. Não houve interação entre tipo de implante e tipo de éster de estradiol para TP IATF nem TP Final. As TP IATF não diferiram entre animais que receberam implante novo ou reutilizado (48,3 vs 48,7%) nem entre os que receberam VE ou BE (49,5 vs 47,5%). O mesmo ocorreu para TP Final segundo tipo de implante, novo ou reutilizado (85,2 vs 86,5%) e segundo tipo de éster de estradiol, VE ou BE (86,5 vs 85,2%). Conclui-se que vacas Nelore amamentando podem apresentar adequadas TP IATF e TP Final quando tratadas por implantes de norgestomet novos ou reutilizados associados indistintamente ao valerato ou benzoato de estradiol. / Pregnacy rates (PR) were compared in Nelore cows (Bos taurus indicus) using new or used norgestomet implants associated to estradiol valerate (EV) and estradiol benzoate (EB). Two hundred forty one Nelore cows were synchronized and divided in two groups, they received auricular implants with 3 mg of norgestomet (n=122) or used norgestomet implants (n=119) previously used for 10 days. The animals that received new or used implants were sub-divided and received an administration (im) of 3 mg of norgestomet plus 5 mg of EV and another group received an administration of 50 mg of progesterone plus 2 mg of EB. When the implant was removed a dose of PGF2&alpha; analogous was given for all animals to assure luteolysis and 24 h later they received (im) of 1,0 mg of EB. The fixed-timed artificial insemination (AIFT) occurred 54-56 h after implant withdrawal. There was no difference between the two implants as well as no diffrence to EV or EB, neither PR AITF nor PR Finish. The PR AIFT did not differ among animals that received that received new or used implants (48,3 vs 48,7%) neither EV nor EB (49,5 vs 47,5%). The same happened to PR Finish either new or used (85,2 vs 86,5%) when EV or EB (86,5 vs 85,2%) were used. In this present study, it was concluded that Nelore suckling cows can show a regulated PR AIFT and PR Finish after treatments with new or used norgestomet implants associated to EV or EB.

Compararam-se as taxas de prenhez (TP) em vacas Nelore (Bos taurus indicus) utilizando implantes de norgestomet novos ou reutilizados associados ao valerato de estradiol (VE) e benzoato de estradiol (BE). Duzentos e quarenta e uma vacas Nelore amamentando foram separadas em dois grupos, e receberam implante auricular contendo 3 mg de norgestomet (n=122) ou implante de norgestomet (n=119) que já havia sido previamente utilizado por 10 dias. Os animais que receberam implante novo ou reutilizado, foram sub-divididos, e um lote recebeu uma injeção (IM) de 3 mg de norgestomet associados a 5 mg de valerato de estradiol e o outro, uma injeção (IM) de 50 mg de progesterona associados a 2 mg de benzoato de estradiol. No momento da retirada do implante todos os animais receberam 1 aplicação de análogo de PGF2&alpha; (IM) para assegurar a luteólise e 24 h após uma injeção de 1,0 mg de BE (IM). A inseminação artificial em tempo fixo (IATF) ocorreu 54-56 h após a retirada do implante. Não houve interação entre tipo de implante e tipo de éster de estradiol para TP IATF nem TP Final. As TP IATF não diferiram entre animais que receberam implante novo ou reutilizado (48,3 vs 48,7%) nem entre os que receberam VE ou BE (49,5 vs 47,5%). O mesmo ocorreu para TP Final segundo tipo de implante, novo ou reutilizado (85,2 vs 86,5%) e segundo tipo de éster de estradiol, VE ou BE (86,5 vs 85,2%). Conclui-se que vacas Nelore amamentando podem apresentar adequadas TP IATF e TP Final quando tratadas por implantes de norgestomet novos ou reutilizados associados indistintamente ao valerato ou benzoato de estradiol. / Pregnacy rates (PR) were compared in Nelore cows (Bos taurus indicus) using new or used norgestomet implants associated to estradiol valerate (EV) and estradiol benzoate (EB). Two hundred forty one Nelore cows were synchronized and divided in two groups, they received auricular implants with 3 mg of norgestomet (n=122) or used norgestomet implants (n=119) previously used for 10 days. The animals that received new or used implants were sub-divided and received an administration (im) of 3 mg of norgestomet plus 5 mg of EV and another group received an administration of 50 mg of progesterone plus 2 mg of EB. When the implant was removed a dose of PGF2&alpha; analogous was given for all animals to assure luteolysis and 24 h later they received (im) of 1,0 mg of EB. The fixed-timed artificial insemination (AIFT) occurred 54-56 h after implant withdrawal. There was no difference between the two implants as well as no diffrence to EV or EB, neither PR AITF nor PR Finish. The PR AIFT did not differ among animals that received that received new or used implants (48,3 vs 48,7%) neither EV nor EB (49,5 vs 47,5%). The same happened to PR Finish either new or used (85,2 vs 86,5%) when EV or EB (86,5 vs 85,2%) were used. In this present study, it was concluded that Nelore suckling cows can show a regulated PR AIFT and PR Finish after treatments with new or used norgestomet implants associated to EV or EB.

In the first experiment, estradiol production was measured in cultured embryos collected on Days 7, 9, 11, 13 and 15 post-mating. Estradiol was detected in Day 7 embryos and production increased approximately 50-fold (P<0.05) in medium recovered from Day 13 compared to Day 11 embryos and was greatest (P<0.05) from Day 15 embryos. The second experiment evaluated the luteotrophic effect of estradiol in the llama. Daily injections of vehicle only, 5 or 10 mg estradiol benzoate (EB) in isopropylmyristate were administered on Days 7-15 (Day 0=hCG ovulation induction). Mean progesterone levels were greater (P<0.05) on Days 14, 15, 16, and 17 in llamas injected with 10 mg EB compared to llamas injected with vehicle or 5 mg EB. The final experiment analyzed differences in estrogen receptor expression in the corpus luteum (CL), ovary, and uterus using Reverse Transcription-Polymerase Chain Reaction. A significant linear regression (P<0.05) was detected for both ER�� (increased from Days 7-11), and ER�� (decreased from Days 7-11) in llama CL. ER�� expression in ovary was higher (P<0.05) compared to Days 7 and 9 CL. ER�� expression in CL was lower on Day 9 (P<0.05) than on Days 7 and 11 in pregnant animals. ER�� expression decreased (P<0.01) from Days 7-11 in uterus and expression was lower in pregnant versus non-pregnant uterus (P<0.10). ER�� expression was higher in pregnant versus non-pregnant uterus (P<0.10). No differences (P>0.10) in ER�� or ER�� expression were detected in endometrium due to reproductive status or days post-mating. A decrease (P<0.01) in ER�� expression was detected in pregnant uterus from Days 7-13. ER�� expression was lower (P=0.12) in juvenile uterus versus pubertal females. ER�� expression in non-pregnant uterus was different (P<0.05) by days post-mating and reproductive status. These data suggest that estradiol produced by the embryo may be involved in embryo migration and maternal recognition of pregnancy in the llama, and these events may be mediated by differential expression of ER subtypes / Graduation date: 2000

published_or_final_version / abstract / toc / Pharmacology / Master / Master of Philosophy

Many studies show that estradiol (E2) and consumption of soy and its primary phytoestrogen component genistein (GEN) can inhibit the formation of colon tumors. However, the effects of E2 and GEN at physiologically relevant levels in non-diseased colonocytes have yet to be investigated. We hypothesized that E2 and GEN could prove to be chemo-protective agents in the colon by moderately increasing apoptosis and decreasing proliferation in a healthy system. Thus, the presented studies focused on evaluating the effects of E2 and GEN in non-malignant colonocytes in vitro and in vivo to determine how the compounds influence the physiology of these cells. E2 (1 nM/L) and GEN treatments (1 and 10 microM/L) decreased cell growth, increased apoptosis, and increased p53 transcriptional activity in young adult mouse colonocytes, a non-malignant cell line. To study further the effects of E2 and GEN in healthy colonic epithelia, we evaluated physiologic changes in colonic crypts in ovariectomized mice given an E2 pellet, 1,000 ppm GEN diet, or a phytoestrogen free diet. As seen in vitro, E2 treated animals had significantly higher rates of apoptosis with GEN trending in the same fashion. These data demonstrate that E2 and GEN alter the physiology of non-malignant colonocytes. Collectively, with our previous data, this suggests that E2 and GEN influence colonocyte physiology and this state may partially explain how these compounds decrease risk of colon cancer.

Thesis (Ph. D.)--Ohio State University, 2007. / Title from first page of PDF file. Includes bibliographical references (p. 212-272).

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 144-155).