Thesis (MA in Religion Studies)--University of South Australia, 1994

Thesis (M. Ed.)--University of Adelaide, Dept. of Education, 1991. / Typescript (Photocopy). Includes bibliographical references (leaves 91-99).

Thesis (D. Min.)--Trinity Evangelical Divinity School, 1994. / Abstract. Includes bibliographical references (leaves 148-156).

Applied research project (D. Min.)--School of Theology and Missions, Oral Roberts University, 2003. / Includes abstract and vita. Includes bibliographical references (leaves 221-232).

Thesis (Ph. D.)--University of Washington, 1999. / Vita. Includes bibliographical references (leaves 148-168).

Thesis (Ph. D.)--University of Chicago, Committee on Human Development, Dept. of Psychology, August 2002. / Includes bibliographical references. Also available on the Internet.

Orientador: Eunice Oba / Banca: Fernanda da Cruz Landim e Alvarenga / Banca: Otavio Mitio Ohashi / Banca: Marcelo Marcondes Seneda / Banca: Mayra Elena Ortiz D'Avila Assumpção / Resumo: Pesquisadores do mundo inteiro têm investigado a função de fatores e eventos biológicos envolvidos na foliculogênese e no processo de maturação ovocitária in vitro em várias espécies, com o intuito de aprimorar tecnologias de produção e manipulação de embriões. O objetivo do presente estudo foi avaliar: a morfometria ovariana; os aspectos morfométricos e ultra-estruturais dos folículos ovarianos; os fatores que influenciam a recuperação, qualidade ovocitária e taxas de maturação nuclear in vitro; as concentrações de progesterona e insulina plasmáticas e os níveis de hormônios esteróides (estradiol e progesterona) e IGF-I no líquido folicular, em búfalas e vacas. Amostras de sangue e os ovários de 86 búfalas (33 gestantes e 53 não gestantes) e de 95 vacas (36 gestantes e 59 não gestantes) foram colhidos em abatedouros. Os ovários foram transportados ao laboratório em solução salina acrescida de penicilina e estreptomicina a 36ºC. As dimensões ovarianas foram mensuradas com o auxílio de paquímetro e balança digitais. Para a maturação ovocitária in vitro, os complexos cumulus oophorus (CCOs) foram recuperados por aspiração folicular, selecionados e cultivados em meio TCM-199 suplementado com 10% de soro fetal bovino, piruvato de sódio, FSH, LH, estradiol, cisteamina e gentamicina, por 22 a 24 horas, a uma temperatura de 38,5ºC, em atmosfera úmida com 5% de CO2 em ar. Para as técnicas de histologia clássica e microscopia eletrônica de transmissão foram processados 10 e 5 ovários de cada espécie, respectivamente. As concentrações de progesterona, insulina, estradiol e IGF-I nas amostras de sangue e fluido folicular foram determinadas por procedimento de radioimunoensaio, utilizando-se kits comerciais. Na análise histológica, verificou-se ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Researchers around the world have investigated the function of biological factors and events during folliculogenisis and in the process of in vitro maturation of oocytes in several species in order to improve technologies of embryo production and manipulation. The aim of the present study was to evaluate: ovarian morphometry; morphological and ultrastructural aspects of ovarian follicles; factors affecting the recovery and competence of oocytes and in vitro nuclear maturation; plasma progesterone and insulin concentrations; progesterone, estradiol and IGF-I in follicular fluid, in bubaline and bovine species. Blood samples and ovaries of 86 buffaloes (33 pregnant and 53 non pregnant) and 95 cows (36 pregnant and 59 non pregnant) were collected at slaughterhouses. The ovaries were transported to the laboratory in saline solution enriched with penicillin and streptomycin at 36ºC. The ovarian dimensions were measured using digital paquimeter and balance. For in vitro maturation process, cumulus-oocyte complexes (COCs) were recovered by follicular aspiration, selected and matured in TCM 199 supplemented with 10% fetal calf serum, sodium pyruvate, LH, FSH, estradiol, gentamicin and cysteamin. In vitro maturation was carried out at 38.5ºC under a controlled gas atmosphere of 5% CO2 in humidified air for 22-24 hours. For classical histology and transmission electron microscopy, 10 and 5 ovaries of each species respectively were processed. Progesterone, insulin, estradiol and IGF-I concentrations of blood and follicular fluid were determined by use of radioimmunoassay and commercial kits. In histological evaluation, the diameters of follicles and oocytes increased according to follicular development. Ultrastructural characterization of bubaline and bovine ovarian follicles showed a differentiated cell apparatus for substrate metabolization, ...(Complete abstract click electronic access below) / Doutor

Made available in DSpace on 2014-06-11T19:35:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-09-23Bitstream added on 2014-06-13T19:05:15Z : No. of bitstreams: 1 tanaka_jlo_dr_sjc.pdf: 558609 bytes, checksum: 1a9708d0868c302c9eb6ce91dd43cc01 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / As alterações anatômicas das vértebras cervicais têm sido empregadas na análise de maturação óssea em Ortodontia e Ortopedia Funcional dos Maxilares. Como essas mudanças são sutis, o objetivo neste estudo foi desenvolver e avaliar um método de análise de maturação óssea empregando-se medidas dessas estruturas. Foram utilizados 246 pares de radiografias cefalométricas laterais e de mão/punho, 135 de indivíduos do sexo feminino e 111 do masculino, divididos em 5 grupos segundo a Curva de Crescimento de Mercadante. Nas radiografias cefalométricas laterais, 15 razões foram obtidas das vértebras C2, C3 e C4, utilizando o programa Radiocef 4.0®. A média de cada razão foi comparada entre os grupos por meio das análises ANOVA e Tukey, que apontaram 10 razões como as que melhor diferem cada fase e que foram utilizadas na criação do novo método. Neste método, denominado Maturação por Razões em Vértebras Cervicais (MRVC), a maturação óssea é classificada em 7 estágios a partir dos resultados das 10 razões. Para sua validação, utilizou-se outra amostra de 58 pares de radiografias cefalométricas laterais e de mão/punho, 28 do sexo feminino e 30 do masculino, submetidas à avaliação de 4 examinadores por três métodos: Curva de Crescimento, Hassel e Farman e método MRVC. Os resultados apontaram melhor desempenho intra e inter-examinadores para a Curva de Crescimento e desempenho semelhante entre o método MRVC e o de Hassel e Farman. Por outro lado, o método MRVC proporcionou resultados próximos ao da Curva de Crescimento na determinação da maturação óssea do indivíduo em relação ao pico do surto de crescimento puberal. Concluiu-se que foi possível criar um método de análise de maturação óssea por medidas das vértebras cervicais em radiografias cefalométricas laterais com eficácia comparável à da Curva de Crescimento. / Anatomic alterations of the cervical vertebrae have been used on the assessment of bone maturation in Orthodontic/Functional orthopedics. Since these changes are subtle, our aim was to create and analyze a method for the assessment of skeletal maturation using measurements of the cervical vertebrae. Two hundred and forty six pairs of lateral cephalometric and hand/wrist radiographs (135 male and 111 female) were used on this study. The sample was separated in 5 groups according to the stage of Mercadante’s Pubertal Growth Curve. Fifteen ratios were calculated with the measurements of C2, C3 and C4 vertebrae. The mean for each ratio was compared among the groups with the ANOVA and Tukey tests, which indicated 10 ratios that better distinguish each stage. These ratios were used on the development of the method Maturation by Ratios on Cervical Vertebrae (MRCV). This method ranks the maturation in 7 stages based on the results of the ratios. For the validation of the method, four examiners assessed 58 pairs of lateral cephalometric and hand/wrist radiographs using 3 methods: Pubertal Growth Curve, Hassel and Farman and MRCV. Better intra and inter-examiner performance were observed for Pubertal Growth Curve method, while the performance with Hassel and Farman and MRCV were almost the same. However, the results for MRCV were close to the Pubertal Growth Curve on the determination of skeletal maturation related to the peak of the pubertal growth. Based on the results, we concluded that it was possible to create a method for skeletal maturation assessment with measurements of the cervical vertebrae in lateral cephalometric radiographs with efficacy comparable to the Pubertal Growth Curve’s.

Made available in DSpace on 2014-06-11T19:35:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-05-05Bitstream added on 2014-06-13T19:05:29Z : No. of bitstreams: 1 leal_ls_dr_botfmvz.pdf: 1424462 bytes, checksum: bbd9bf26b7adb5dc3b7bb266434ac12a (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Pesquisadores do mundo inteiro têm investigado a função de fatores e eventos biológicos envolvidos na foliculogênese e no processo de maturação ovocitária in vitro em várias espécies, com o intuito de aprimorar tecnologias de produção e manipulação de embriões. O objetivo do presente estudo foi avaliar: a morfometria ovariana; os aspectos morfométricos e ultra-estruturais dos folículos ovarianos; os fatores que influenciam a recuperação, qualidade ovocitária e taxas de maturação nuclear in vitro; as concentrações de progesterona e insulina plasmáticas e os níveis de hormônios esteróides (estradiol e progesterona) e IGF-I no líquido folicular, em búfalas e vacas. Amostras de sangue e os ovários de 86 búfalas (33 gestantes e 53 não gestantes) e de 95 vacas (36 gestantes e 59 não gestantes) foram colhidos em abatedouros. Os ovários foram transportados ao laboratório em solução salina acrescida de penicilina e estreptomicina a 36ºC. As dimensões ovarianas foram mensuradas com o auxílio de paquímetro e balança digitais. Para a maturação ovocitária in vitro, os complexos cumulus oophorus (CCOs) foram recuperados por aspiração folicular, selecionados e cultivados em meio TCM-199 suplementado com 10% de soro fetal bovino, piruvato de sódio, FSH, LH, estradiol, cisteamina e gentamicina, por 22 a 24 horas, a uma temperatura de 38,5ºC, em atmosfera úmida com 5% de CO2 em ar. Para as técnicas de histologia clássica e microscopia eletrônica de transmissão foram processados 10 e 5 ovários de cada espécie, respectivamente. As concentrações de progesterona, insulina, estradiol e IGF-I nas amostras de sangue e fluido folicular foram determinadas por procedimento de radioimunoensaio, utilizando-se kits comerciais. Na análise histológica, verificou-se... / Researchers around the world have investigated the function of biological factors and events during folliculogenisis and in the process of in vitro maturation of oocytes in several species in order to improve technologies of embryo production and manipulation. The aim of the present study was to evaluate: ovarian morphometry; morphological and ultrastructural aspects of ovarian follicles; factors affecting the recovery and competence of oocytes and in vitro nuclear maturation; plasma progesterone and insulin concentrations; progesterone, estradiol and IGF-I in follicular fluid, in bubaline and bovine species. Blood samples and ovaries of 86 buffaloes (33 pregnant and 53 non pregnant) and 95 cows (36 pregnant and 59 non pregnant) were collected at slaughterhouses. The ovaries were transported to the laboratory in saline solution enriched with penicillin and streptomycin at 36ºC. The ovarian dimensions were measured using digital paquimeter and balance. For in vitro maturation process, cumulus-oocyte complexes (COCs) were recovered by follicular aspiration, selected and matured in TCM 199 supplemented with 10% fetal calf serum, sodium pyruvate, LH, FSH, estradiol, gentamicin and cysteamin. In vitro maturation was carried out at 38.5ºC under a controlled gas atmosphere of 5% CO2 in humidified air for 22-24 hours. For classical histology and transmission electron microscopy, 10 and 5 ovaries of each species respectively were processed. Progesterone, insulin, estradiol and IGF-I concentrations of blood and follicular fluid were determined by use of radioimmunoassay and commercial kits. In histological evaluation, the diameters of follicles and oocytes increased according to follicular development. Ultrastructural characterization of bubaline and bovine ovarian follicles showed a differentiated cell apparatus for substrate metabolization, ...(Complete abstract click electronic access below)

The ability of oocytes and early cleavage-stage embryos to regulate their volume is essential to avoid developmental arrests at in vivo-osmolarities. This is accomplished primarily via GLYT1-mediated glycine transport into the cells. GLYT1 activity has previously been shown to be absent in freshly isolated oocytes but becomes activated ~3-4 hours after oocyte maturation has been initiated either by isolation from ovarian follicles in vitro or following an ovulatory stimulus in vivo. GLYT1 activity then persists until the 4-cell stage of preimplantation embryo development. GLYT1 has been shown to spontaneously activate in oocytes that are isolated from follicles either as denuded oocytes or as cumulus-oocyte complexes (COCs), this implies that GLYT1 activity is suppressed in intact follicles in the ovary. However, it is not known how GLYT1 activity is suppressed within the ovarian follicle or how initial GLYT1 activation occurs. The activation of independent cell volume regulation in oocytes first involves the release of the strong adhesion between the oocyte and zona pellucida (ZP) followed by secondary GLYT1 activation. These two processes have been shown to occur spontaneously in fully grown oocytes following isolation from ovarian follicles, however, it is not known whether small growing oocytes within ovarian follicles already possess the ability to detach from the ZP and activate GLYT1. An osmotic assay was used to determine when during oogenesis oocytes are first able to detach from the ZP while the ability to activate GLYT1 was determined by measuring [3H]-glycine uptake into oocytes. I found that oocytes acquire the ability to detach from the ZP when they are nearly fully grown and similarly, that high levels of GLYT1 activity first develop in isolated oocytes during the late stages of oogenesis. Furthermore, I showed that SLC6A9 protein (GLYT1 transporter protein) and Slc6a9a transcripts steadily increased during oogenesis with SLC6A9 protein becoming localized to the oocyte plasma membrane during oocyte growth with predominant membrane localization apparent in fully grown oocytes. Taken together, these results suggest that oocytes become able to detach from the ZP and fully activate GLYT1 towards the end of oogenesis but that these processes remain suppressed in the ovarian follicle. Intact and punctured antral follicles were used as a model to examine the potential mechanism(s) mediating GLYT1 suppression before ovulation is triggered. Using these models, I found that GLYT1 activity remains suppressed within preovulatory antral follicles in contrast to the spontaneous GLYT1 activation that occurred in isolated denuded oocytes or within COCs. Recently, the mechanism mediating oocyte maintenance of prophase I arrest within the ovarian follicle was elucidated and was shown to depend on the release of Natriuretic Peptide Precursor C (NPPC) from mural granulosa cells (MGCs) into follicular fluid which binds to NPR2 guanylate cyclases on cumulus cells stimulating the production of cyclic GMP (cGMP) within these cells. Diffusion of cGMP from cumulus granulosa cells to the oocyte via gap junctions is required to maintain meiotic arrest. Although GLYT1 activation and meiotic resumption are both suppressed in antral follicles prior to the ovulatory trigger and these two processes occur simultaneously following oocyte isolation from ovaries, I have shown here that GLYT1 suppression within the preovulatory antral follicle is mediated by a mechanism distinct from the gap junction-dependent NPPC-cGMP pathway controlling meiotic arrest. I also showed for the first time a direct requirement for meiotic arrest of both gap junctions between granulosa cells (composed of connexin-43) and between the inner layer of cumulus granulosa cells and the oocyte (composed of connexin-37). Since I showed that GLYT1 was suppressed in isolated antral follicles but not COCs, I hypothesized that MGCs are required to maintain low GLYT1 activity in antral follicles. I showed here that MGCs isolated from preovulatory antral follicles were sufficient to maintain GLYT1 suppression in co-cultured COCs, but not denuded oocytes. Furthermore, I found that GLYT1 activity was suppressed in COCs cultured in conditioned medium from MGC cultures. Thus, GLYT1 activity appears to be suppressed within the ovary prior to the ovulatory LH-stimulus likely by an unidentified inhibitory signal within the ovarian follicle originating from the MGCs and propagated by a gap junction-independent mechanism involving multiple cell types in the follicle.