Thesis (M. Phil.)--University of Hong Kong, 2009. / Includes bibliographical references (leaves 182-187) Also available in print.

Thesis (Ph. D.)--University of Missouri-Columbia, 1996. / Typescript. Vita. Includes bibliographical references (leaves 136-172). Also available on the Internet.

Thesis--Copenhagen. / Summary in Danish. Bibliography: p. 219-235.

Thesis--Copenhagen. / Summary in Danish. Bibliography: p. 219-235.

Thesis (M.Sc.)--Acadia University, 2000. / Includes bibliographical references (leaves 116-127). Also available on the Internet via the World Wide Web.

Thesis (Ph. D.)--University of Missouri-Columbia, 1996. / Typescript. Vita. Includes bibliographical references (leaves 136-172). Also available on the Internet.

Thesis (Ph. D.)--University of Washington, 1996. / Vita. Includes bibliographical references (leaves [73]-85).

Background and purpose: Rhizoma Chuanxiong (CX), the dry rhizome of Ligusticum chuanxiong Hort., is a commonly used Chinese herbal medicine to treat gynecological diseases. So far, more than 60 chemical components have been identified from CX such as volatile oils (ligustilide, etc.), phenolic acids (ferulic acid, etc.) and alkaloids (chuanxiongzine, etc.). These components in CX are the basis of its wide pharmacodynamic actions including estrogen-like, progesterone-like and anti-coagulant/anti- platelet effects. In our recent survey based on previous published clinical trials, CX was ranked as one of the top 20 herbs commonly used for anti-miscarriages amongst Chinese pregnant women. However, CX should be used with caution during pregnancy as its property of 2invigorating blood circulation and removing blood stagnation3. Despite its wide applications, the safe dosage of CX in pregnant women remains unclear with no records found in the current Chinese Pharmacopoeia or other guidelines. Thus, verification regarding the impacts of CX preparations and its components in embryonic development is urgently required. In view of the limited experimental evidence that is currently available to assess the safety of CX, this project aims to (1) identify the general impacts of CX aqueous extract in maternal function and fetal development with an in vivo mouse model; and to (2) investigate the adverse impacts and underlying mechanisms of CX aqueous extract in fetal bone development with a biomarker assay and metabolomics analysis.;Concusion: CX aqueous extract at a low dosage of 2 g/kg/day (equals to the daily dosage of human adults) did not cause adverse effect in pregnant mice, and it suggested that this dosage of CX preparations should be safe for pregnant women. Our data demonstrated that high dosage and long-term use of CX aqueous extract might result in embryonic toxicities including fetal bone malformations for the first time. As the CX aqueous extract in this study was not contaminated by pesticide residues and heavy metals, the adverse impacts of CX aqueous extract should be considered as a result of its intrinsic components in the herb. Furthermore, CX aqueous extract might significantly down-regulate biomarkers related to bone formation and metabolism during osteogenesis. It is therefore valuable to establish a practical approach to systematically assess the safety of CX and other herbal medicines.;Method: Referred to the guidelines of WHO, the Chinese Pharmacopoeia and the Hong Kong Chinese Materia Medica Standards, CX aqueous extract was prepared, and its reference marker (ligustilide and ferulic acid) were quantitatively authenticated by HPLC analysis. LC/MS fingerprint analysis was performed for the quality control purposes. In addition, pesticide residues and heavy metals found in CX aqueous extract were examined using GC-MS and ICP-MS analysis. In the Segment II study as per FDA and OECD guidelines, pregnant mice were randomly assigned into 6 groups (n=18 per group): i.e. mice were orally administrated with distilled water as the negative controls (Group 1); or CX aqueous extract of 2, 16, 24 and 32 g/kg/day respectively from the gestation day (GD)6 to 16(Group 2, 3, 4 and 5); or vitamin A (200,000 IU) on GD7, 9 and 11 as the positive controls (Group 6). All mice were sacrificed to assess maternal and fetal parameters on the GD18. In the mechanistic study, the expressions of biomarkers related to fetal bone development including PICP, ICTP, B-ALP, BGP, Gdf-5, BMPs, BMP-6, BMP-8, BMP-11, IL-4, IL-4r, IL-10 and IL-10r in fetal tissue samples of the Group 1 and 5 (32 g/kg/day, n=18) were measured using ELISA analyses on GD16. Meanwhile, the metabolites of two-group samples were also analyzed by the UHD Accurate-Mass Q- TOF LC/MS, and profiling data was further analyzed by specific software. During statistical analysis, measurement data from G1, 2, 3 4 and 5 groups were analyzed using one-way ANOVA(SPSS software, version 16.0). LSD test in Post hoc method was applied to compare differences between every two groups. Pearsons x 2 - test was used to analyze category data from G1, 2, 3, 4 and 5 groups, and Fishers exact test was applied to compare differences between different groups. The student t-test was also used to compare differences between G1 and G6 groups in animal studies as well as G1 and G5 groups using ELISA or metabolomics results. An intragroup difference with a p-value less than 0.05 was considered as statistical significant.;Resutt:(1) There was no statistical significant difference in maternal and fetal parameters found between the Group 1 and 2 (p> 0.05). However, the maternal body weight (BW), gravid uterine weight, corrected BW change, live fetus/litter, mean fetal BW in the Group 4 and 5 were significantly lower than those in the Group 1(p

Skeletal muscle is a highly metabolically active tissue, both in the adult and the fetus. Mitochondria are essential in providing energy in the form of ATP from the oxidative metabolism of carbohydrates, fats and amino acids. Mitochondrial function is influenced by the abundance and activity of the complexes comprising the electron transfer system (ETS) and the balance between mitochondrial fusion and fission. Any factors which affect the development of skeletal muscle, and mitochondria in particular, may have an impact not only on neonatal health but also on the metabolic health of the adult offspring. However, the normal developmental profile of skeletal muscle mitochondrial function as the fetus prepares for the increased metabolic challenges associated with extrauterine life, is not well characterised. The hormones, cortisol and triiodothyronine (T3) are known to be crucial in the maturation of several physiological processes during late gestation. Further, their role in regulating adult metabolism is well-documented. However, whether they play a role in regulating fetal mitochondrial function is unknown. Using fetal sheep, the aims of this project were twofold: 1) to determine any changes in skeletal muscle mitochondrial function which occur over the last third of gestation and in the first two days of neonatal life and 2) to determine any regulatory roles of cortisol and T3 in these developmental changes. Mixed fibre-type skeletal muscle was collected from fetuses at 3 time points over late gestation and from newborn lambs. In addition, skeletal muscle samples were taken from fetuses which had been thyroidectomised (TX) and fetuses infused with either T3 or cortisol. Respirometry, enzyme assays, qRT-PCR and western blotting were carried out on the skeletal muscle samples in order to assess mitochondrial parameters. Mitochondrial activity, as measured by carbohydrate- and fat- stimulated ADP-coupled oxygen uptake, increased with age in a thyroid hormone dependent manner, rising predominantly postnatally. Mitochondrial density, abundance of ETS complexes I-IV and ATP-synthase and expression of the adenine nucleotide transferase 1 and mitofusin 2 were all positively influenced by age, with the natural prepartum rise being prevented in the thyroidectomised fetuses. However, T3 infusion alone was insufficient to raise any of these factors prematurely. Cortisol infusion resulted in an increase in some aspects of mitochondrial oxidative capacity in a muscle-specific manner. Overall, the data presented shows that there are developmental changes in skeletal muscle mitochondria during the perinatal period. They also suggest that these changes are regulated by both cortisol and thyroid hormones in preparation for birth, although neither hormone alone was sufficient to induce all the functional changes.

Resumo: Objetivou-se testar diferentes protocolos de cultivo in vitro e in vivo de folículos ovarianos pré-antrais de fetos bovinos. Para tanto, um total de 41 ovários de fetos bovinos foram obtidos em matadouro, transportados e utilizados para o cultivo in vitro (n=20) e para o xenotransplante (n=21). Após processados no laboratório em fragmentos entre 0,5 e 1 mm3 foram encaminhados para os cultivos O cultivo in vitro baseou-se em protocolo bem sucedido de cultivo de FOPA em caprinos e testou diferentes fontes de macromoléculas e a utilização do azul de tripan na viabilidade dos tecidos cultivados a uma atmosfera controlada de 5% CO2 em ar, a 38,5°C e nutridos com dMEM (300 mOsm/L, pH 7,2) suplementado com antibióticos, ITS, piruvato de sódio, glutamina, hipoxantina, dAMPc, bFSH e IGF-I. A depender do tratamento, foi adicionado BSA (0,1%) SFB (10%) ou PVA (1%). O cultivo in vivo de FOPA foi executado por xenotransplante sob a cápsula renal num total de 65 camundongas imunodeficientes. Desenvolveu-se uma técnica de biopsia e verificou-se o efeito do tempo de transplante (30, 60 e 30 e 60 dias após o transplante) sobre a percentagem e a viabilidade de FOPA bem como a possível presença de folículos antrais. Num segundo momento, 32 receptoras receberam estímulo hormonal de 10 UI de eCG (n=18) e 10 UI r-hFSH (n=14). Os resultados mostraram que o cultivo com PVA apresentou FOPA normais em percentagem semelhante aos cultivos com BSA e PVA. Quanto ao cultivo por xenotransplante, observou-se o crescimento sucessivo de FOPA até estádios antrais ao longo do tempo de transplante (> 30 dias). O resultado das estimulações exógenas apresentou folículos antrais com oócitos que apresentaram o cumulus expandido em 2/5 (40%) dos oócitos selecionados para MIV de cada um dos tratamentos propostos. Concluindo, FOPA oriundos de fetos bovinos podem ser cultivados por pelo menos 8 dias em PVA... (Resumo completo clicar acesso eletrônico abaixo) / Abstract: This study aimed to evaluate different protocols of in vitro and in vivo preantral follicles (PFs) culture from bovine fetus. Thus, a total of 41 fetal bovine ovaries, from a slaughterhouse, were collected and transported at laboratory for in vitro culture (n=20) and for xenotransplantation (n=21), after processing into small cortical pieces measuring between 0,5 and 1mm3 the slices were cultured. The in vitro culture was based on well successful protocol of preantral follicles in caprine and tested different sources of macromolecules and trypan blue viability of cultured tissues cultured at controlled atmosphere (5%CO2 in air, 39°C). The culture medium used was dMEM (300 mOsm/L, pH 7, 2) supplemented with antibiotics, ITS, sodium pyruvate, glutamine, hypoxanthine, dAMPc, bFSH, and IGF-I. Depending of treatment, was added BSA (0,1%), FCS (10%) or PVA (1%). In vivo culture of preantral follicles was carried out by xenotransplantation under renal capsule of immunodeficient females mice (total of 65) that were submitted to a biopsy technique previously developed for tissue collection and to verify the effectiveness of time of transplantation (30, 60 and 30 and 60 days post surgery) under percentage and viability as well as putative growth of PFs to antral follicles. At second stage, 32 recipient mice were submitted to hormonal stimuli with 10 IU of eCG (n=18) and 10 IU of r-hFSH (n=14). The results showed that PVA culture presented normally PF in similar distribution when compared with BSA and PVA culture. Regarding xenotransplantation, successive growth of PF until antral stages was observed belong time of transplantation. Exogenous stimulation presented oocytes with expanded cumulus in 2/5 (40%) of selected oocytes for IVM from each treatment. Conclusively PFs from fetal bovine ovaries can be cultured at PVA at least 8 days and grows until antral stages after xenotransplantation procedures... (Complete abstract click electronic access below) / Orientador: Maria Rita Pacheco / Coorientador: Wilter Ricardo Russiano Vicente / Banca: José Ricardo de Figueiredo / Banca: José Antonio Visintin / Banca: Joaquim Mansano Garcia / Banca: Paulo Henrique Franceschini / Doutor