Antral ovarian folliculogenesis involves recruitment of a cohort of small follicles, physiological selection of a dominant follicle, and ovulation. The mechanism of selection has not been precisely determined. Identification of the timing of preovulatory selection is a key component in understanding natural and peri-menopausal ovarian function, ovarian suppression for contraception, and improvement of ovarian stimulation protocols. Morphologic characteristics obtained by ultrasonography cannot be precisely quantitated by the human eye. Computer-assisted image analysis overcomes subjective human evaluation of ultrasonographic images.<p> The objectives of this research were to assess ultrasound image attributes of human dominant (DF) and 1st subordinate (SF1) ovarian follicles during natural menstrual cycles and following discontinuation of conventional and continuous oral contraceptives (OC). We utilized sophisticated computer algorithms to elucidate an association between image attributes and physiologic status of follicles. Transvaginal ultrasonographic images obtained in 2 previous studies were used to quantify changes that occur in ovarian follicles.<p> We detected quantitative differences between the dominant and largest subordinate follicles of ovulatory and major anovulatory follicular waves, as well as during the first wave following OC discontinuation. Differences in ultrasonographic image attributes were associated with the physiological status of follicles. Evidence of follicular dominance in follicles which develop during major ovulatory waves or following OC discontinuation can be detected prior to the time of selection manifest by differences in dominant and subordinate follicle diameters. In addition, differences in quantitative image attributes were detected between ovulatory and anovulatory DF. Follicles that develop following conventional and continuous OC administration schemes exhibit the same image characteristics.<p> Further research is necessary to elucidate the exact correlation of follicle image attributes during all stages of development with histological characteristics, prediction of the timing of DF selection and the effects of different OC formulations on follicle development during and following OC cessation. Computer-assisted image analysis of ultrasound images has the potential to develop into a diagnostic, prognostic, and research tool for the in vivo evaluation of ovarian physiology and pathology and elucidate biologically important times such as physiologic selection, ovulation of DF and characterization of abnormal follicles (i.e., follicular cysts, luteinized unovulated follicles).

Bibliography: leaves 87-113. / xi, 113, [72] leaves, [42] leaves of plates : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Presents results from an investigation into the regulation of hair-specific gene expression, including attempts to produce an immortalised hair follicle cortical cell line for this purpose and the use of mouse transgenesis and invitro gel mobility shift assays. / Thesis (Ph.D.)--University of Adelaide, Dept. of Biochemistry, 1995

This study was designed to determine the effects of stage of lactation and subsequent energy status on metabolic and endocrine measures, follicular development, and the quality of oocytes obtained from Holstein cows. Holstein cows were selected prior to calving and assigned to the early lactation (EL) group (n=8) while, cows at d 90 postpartum were selected for the mid-lactation (ML) group (n=7). Blood samples were taken twice weekly from 4 wk prior to the start of follicular aspirations and then on through the aspiration periods for metabolite and hormone determination. Ultrasound-guided transvaginal follicular aspiration (TVFA) was conducted twice weekly for a 10-wk period on all cows. Follicular fluid samples were obtained from the largest follicle, > 10 mm in diameter, for hormone determination. All data were analyzed by ANOVA, using the general linear model procedures. Mean energy balance was positive for (2.43 ± 0.32 Mcal/kg) for ML cows and negative (-1.55 ± 0.33 Mcal/kg) for EL cows. In ML cows serum progesterone (P4) decreased rapidly from 2.7 ± 0.1 ng/ml at the first aspiration session to a nadir of 0.33 ± 0.1 ng/ml at wk 8, while follicular fluid P4 increased from 0.9 ± 0.5 to 5.6 ± 0.5 ng/ml. In the EL cows serum and follicular fluid P4 remained relatively constant over the course of aspirations. There was a linear increase in follicular fluid insulin-like growth factor I (IGF-I) for EL and ML cows, however the increase was more rapid for ML cows (159 ± 36 to 200 ± 36 ng/ml) than for EL cows (145 ± 36 to 164 ± 36 ng/ml). Over the aspiration period nonesterified fatty acids (NEFA) declined rapidly for the EL cows (0.32 ± 0.2 to 0.22 ± 0.2 mEq/L), while serum NEFA for the ML cows were relatively stable (0.19 ± 0.2 to 0.22 ± 0.2 mEq/L). The number of follicles observed during the aspiration sessions increased linearly for both EL and ML cows (P < 0.05) over the 10-wk period. However, the increase was larger for the ML cows than for the EL cows, going from 14.2 ± 0.5 to 18.1 ± 0.5 and 14.9 ± 0.3 to 15.7 ± 0.5, respectively. These results show that cows in early lactation are physiologically under more production stress than cows in mid lactation. Furthermore, increasing levels of serum and follicular fluid IGF-I in mid lactation may reflect differences in follicle and oocyte measures. / Master of Science

Lack of hair due to alopecia or skin grafting procedures causes significant distress due to hair's role in social and sexual communication. Only limited pharmacological agents are currently available to stimulate hair growth; their development is hampered by inappropriate model systems. Most research involves large terminal scalp follicles rather than the clinical targets of tiny vellus or intermediate follicles. The overall aim of this thesis was to develop a novel model system based on intermediate hair follicles. Initially, intermediate follicles from female pre-auricular skin were characterised and compared to matched terminal follicles. Intermediate follicles were smaller, less pigmented, shorter and possessed a more 'tubular' bulb morphology than their more 'bulbous' terminal counterparts. Significant correlations were demonstrated between various hair follicle measurements and corresponding dermal papilla diameters. Isolated terminal follicles grew significantly more than intermediate hair follicles in organ culture for 9 days. Testosterone (10nM), the major regulator of human hair growth, increased only intermediate follicle growth; the anti-androgen, cyproterone acetate (1¿M), prevented this stimulation, unlike the 5¿-reductase type 2 inhibitor finasteride (40ng/ml). Immunohistochemistry demonstrated androgen receptor and 5¿-reductase type 2 proteins in both follicle types, while quantitative real-time PCR and gene microarray analysis detected their increased gene expression in intermediate follicles. Thus, smaller intermediate follicles showed major morphological and gene expression differences to terminal follicles in vivo and retained significant, biologically-relevant differences in vitro in organ culture including androgen-responsiveness. Therefore, intermediate hair follicles offer a novel, exciting, more clinically relevant, albeit technically difficult, model for future investigations into hair growth.

No description available.

Os répteis compõem hoje uma classe com mais de 8.000 espécies e em razão das restrições na importação desses animais, o risco na introdução de doenças exóticas, o crescente número de espécies ameaçadas no mundo, entre outros, a reprodução e a manutenção em cativeiro desses animais é muito importante. No Sul da Califórnia, que é considerado um Hotspot para a biodiversidade, a Zoological Society of San Diego mantém uma reserva ecológica com aproximadamente 400 hectares e formada por vegetação tipo Chaparral, Coastal Sage Scrub e áreas cobertas por cactos (Opuntia sp.). Durante o período de junho de 2005 a julho de 2006 foram capturadas 96 cascavéis das espécies Crotalus ruber ruber, C. oreganus helleri e C. mitchellii phyrrus, durante os trabalhos de campo pela procura visual limitada por tempo. Foram realizadas avaliações morfológicas e reprodutivas em todas essas serpentes capturadas. Nos machos realizou-se a colheita e avaliação de sêmen, incluindo duas colorações específicas para avaliar integridade de membrana espermática e do acrossoma. Nas fêmeas foi realizada a avaliação ultra-sonográfica dos ovários, com mensuração e contagem de folículos, além do diagnóstico de gestação. Para a C. ruber ruber, a serpente mais abundante neste estudo, foram observadas fêmeas prenhes no verão e vitelogênicas na primavera, outono e inverno. Apenas as fêmeas desta espécie com condição corporal boa ou muito boa apresentaram-se vitelogênicas ou prenhes. Nos machos dentre as características seminais avaliadas, não foi observada a diferença estatisticamente significante ao longo das estações do ano. Em paralelo a este estudo, também foram testados protocolos para a congelação de sêmen de serpentes, com a obtenção de resultados promissores, discutidos em capítulo específico. As informações obtidas neste estudo visam contribuir para a compreensão da biologia reprodutiva e conservação das serpentes, em cativeiro ou vida-livre. / There are more than 8,000 reptile species in world. Today´s restrictions to importation of these species around world, the risk of introduction of exotic diseases, the growing number of endangered reptiles, among other reasons, emphasize the importance of reproduction in captivity and ex-situ conservation. South California is a Hotspot for biodiversity and the Zoological Society of San Diego maintains a nature preserve in Escondido City. In an area of about 400 hectares, composed by vegetation types like Chaparral and Coastal Sage Scrub, it has been found the greatest diversity of snake in region. Between May 2005 and July 2006, 96 rattlesnakes of species Crotalus ruber ruber, C. oreganus helleri and C. mitchellii phyrrus were captured by active searching during field work. In the Laboratory of Reproductive Physiology the snakes were measured, weighted, and marked the captured snakes. We evaluated their body condition, collected and evaluated semen from males and performed ultrasound exams in females. Snakes were released up to 24 hours after capture. The Crotalus ruber ruber studied initiated vitellogenesis in fall, courtship and mating occurred in spring and gravid snake were observed in spring and summer. Only females with a good or very good body condition were in secondary vitellogenesis or gravid. There were no statistical differences in seminal parameters among season in males of the Crotalus ruber ruber. Protocols to freeze snake´s semen were tested in this study, with positive results that are discussed in specific chapter. This study results contribute to understand of snake´s biology of reproduction and in or ex-situ conservation.

Os répteis compõem hoje uma classe com mais de 8.000 espécies e em razão das restrições na importação desses animais, o risco na introdução de doenças exóticas, o crescente número de espécies ameaçadas no mundo, entre outros, a reprodução e a manutenção em cativeiro desses animais é muito importante. No Sul da Califórnia, que é considerado um Hotspot para a biodiversidade, a Zoological Society of San Diego mantém uma reserva ecológica com aproximadamente 400 hectares e formada por vegetação tipo Chaparral, Coastal Sage Scrub e áreas cobertas por cactos (Opuntia sp.). Durante o período de junho de 2005 a julho de 2006 foram capturadas 96 cascavéis das espécies Crotalus ruber ruber, C. oreganus helleri e C. mitchellii phyrrus, durante os trabalhos de campo pela procura visual limitada por tempo. Foram realizadas avaliações morfológicas e reprodutivas em todas essas serpentes capturadas. Nos machos realizou-se a colheita e avaliação de sêmen, incluindo duas colorações específicas para avaliar integridade de membrana espermática e do acrossoma. Nas fêmeas foi realizada a avaliação ultra-sonográfica dos ovários, com mensuração e contagem de folículos, além do diagnóstico de gestação. Para a C. ruber ruber, a serpente mais abundante neste estudo, foram observadas fêmeas prenhes no verão e vitelogênicas na primavera, outono e inverno. Apenas as fêmeas desta espécie com condição corporal boa ou muito boa apresentaram-se vitelogênicas ou prenhes. Nos machos dentre as características seminais avaliadas, não foi observada a diferença estatisticamente significante ao longo das estações do ano. Em paralelo a este estudo, também foram testados protocolos para a congelação de sêmen de serpentes, com a obtenção de resultados promissores, discutidos em capítulo específico. As informações obtidas neste estudo visam contribuir para a compreensão da biologia reprodutiva e conservação das serpentes, em cativeiro ou vida-livre. / There are more than 8,000 reptile species in world. Today´s restrictions to importation of these species around world, the risk of introduction of exotic diseases, the growing number of endangered reptiles, among other reasons, emphasize the importance of reproduction in captivity and ex-situ conservation. South California is a Hotspot for biodiversity and the Zoological Society of San Diego maintains a nature preserve in Escondido City. In an area of about 400 hectares, composed by vegetation types like Chaparral and Coastal Sage Scrub, it has been found the greatest diversity of snake in region. Between May 2005 and July 2006, 96 rattlesnakes of species Crotalus ruber ruber, C. oreganus helleri and C. mitchellii phyrrus were captured by active searching during field work. In the Laboratory of Reproductive Physiology the snakes were measured, weighted, and marked the captured snakes. We evaluated their body condition, collected and evaluated semen from males and performed ultrasound exams in females. Snakes were released up to 24 hours after capture. The Crotalus ruber ruber studied initiated vitellogenesis in fall, courtship and mating occurred in spring and gravid snake were observed in spring and summer. Only females with a good or very good body condition were in secondary vitellogenesis or gravid. There were no statistical differences in seminal parameters among season in males of the Crotalus ruber ruber. Protocols to freeze snake´s semen were tested in this study, with positive results that are discussed in specific chapter. This study results contribute to understand of snake´s biology of reproduction and in or ex-situ conservation.

Molecular and behavioural characterisation of ovine dermal papilla cells performed in this study indicate they synthesise a highly specialised extracellular matrix (ECM). This is conserved between different species and distinguishes papilla cells from dermal fibroblasts with which they have a common origin. The composition of the dermal papilla ECM is temporally and spatially regulated during wool follicle development. It was shown that the ECM associated with dermal papilla cells in foetal sheep skin becomes specialised in regard to chondroitin sulphate synthesis prior to the appearance of follicle primordia. Chrondroitin sulphate and fibronectin were present in the ECM of dermal papilla cells throughout follicle development and during fibre production. Cellular differentiation antigen 44 was present in the ECM od papilla cells exclusively during the formation of dermal papilla, while laminin was present in the dermal papilla ECM of fibre producing follicles only. Co-operation between chondroitin sulphate, fibronectin, and CD44 in regulating the agrregative and proliferative behaviour of papilla cells was demonstrated in culture. Finally, the inhibition of proteoglycan synthesis in newborn mouse skin was found to disrupt the growth of existing follicles and the generation of new ones. Together these findings demonstrate that chondroitin sulphate is intimately associated with the earliest interactions between epithelial and mesenchymal cells during the formation of follicle primordia. It is likely that the interactions specifically involve fibronectin and CD44, and possibly other ECM molecules which have he effect of regulating the behaviour of papilla cells / Doctor of Philosophy (PhD)

Two studies were designed to characterize the effects of a non-steroidal aromatase inhibitor, letrozole, on ovarian function in cattle. The specific objective was to test the hypothesis that letrozole will arrest dominant follicle growth resulting in emergence of a new follicular wave at a predictable interval post-treatment. In a first experiment, postpubertal beef heifers were assigned randomly to four treatment groups and given phosphate-buffered saline (controls; n=10), or letrozole at a dose of 500 (n=9), 250 (n=10), or 125 (n=10) µg/kg intravenously 4 days after follicular ablation (~2.5 days after wave emergence). In a second study, postpubertal beef heifers were assigned randomly to four treatment groups. One group received no treatment (control; n=17) and the other groups (n=9-10) were given 85 µg/kg of letrozole per day (250 µg/kg total dose), from Days 1 to 3, Days 3 to 5, or Days 5 to 7 (Day 0 = pre-treatment ovulation,) corresponding to the periods before, during and after selection of the dominant follicle, respectively. Follicular dynamics were monitored ultrasonically and blood samples were collected for endocrine assays. Follicle diameter profiles and plasma LH, FSH, and estradiol concentrations were analyzed. Additionally, during the second trial, CL diameter profiles and plasma progesterone concentrations were measured. In both studies, the diameter profile of the dominant follicle was larger in heifers treated with letrozole than in control heifers (P<0.05) and the intervals to new wave emergence and onset of regression of the extant dominant follicle were longer (P<0.05) in heifers treated with letrozole than in controls, regardless of the dose (high, medium, or low; single vs multiple) and the stage of the follicle wave in which treatments were initiated. Furthermore, during the second experiment, the mean CL diameter was larger in letrozole-treated heifers, although there were no differences in plasma progesterone concentrations between treated and control animals. The effects on dominant follicle and CL diameter profiles appeared to be the result of the significantly increased plasma LH concentrations observed in letrozole-treated animals during both treatment approaches (single vs multiple dose). Incomplete and inconsistent inhibition of estradiol production and the lack of a surge on FSH observed in both experiments may be a result of insufficient circulating levels of letrozole during the treatment period. In summary, a single or multiple dose of letrozole did not induce regression of the extant dominant follicle, nor did it directly affect FSH release. Conversely, letrozole extended the lifespan of the dominant follicle, in association with increased endogenous levels of LH, thereby delaying the next FSH surge and subsequent follicular wave emergence. Results suggest that letrozole has potential as a non-steroidal method for controlling ovarian function in cattle, but further studies are warranted to clarify dosage and timing of treatment to predictably affect follicular wave dynamics in cattle.

Two studies were designed to characterize the effects of a non-steroidal aromatase inhibitor, letrozole, on ovarian function in cattle. The specific objective was to test the hypothesis that letrozole will arrest dominant follicle growth resulting in emergence of a new follicular wave at a predictable interval post-treatment. In a first experiment, postpubertal beef heifers were assigned randomly to four treatment groups and given phosphate-buffered saline (controls; n=10), or letrozole at a dose of 500 (n=9), 250 (n=10), or 125 (n=10) µg/kg intravenously 4 days after follicular ablation (~2.5 days after wave emergence). In a second study, postpubertal beef heifers were assigned randomly to four treatment groups. One group received no treatment (control; n=17) and the other groups (n=9-10) were given 85 µg/kg of letrozole per day (250 µg/kg total dose), from Days 1 to 3, Days 3 to 5, or Days 5 to 7 (Day 0 = pre-treatment ovulation,) corresponding to the periods before, during and after selection of the dominant follicle, respectively. Follicular dynamics were monitored ultrasonically and blood samples were collected for endocrine assays. Follicle diameter profiles and plasma LH, FSH, and estradiol concentrations were analyzed. Additionally, during the second trial, CL diameter profiles and plasma progesterone concentrations were measured. In both studies, the diameter profile of the dominant follicle was larger in heifers treated with letrozole than in control heifers (P<0.05) and the intervals to new wave emergence and onset of regression of the extant dominant follicle were longer (P<0.05) in heifers treated with letrozole than in controls, regardless of the dose (high, medium, or low; single vs multiple) and the stage of the follicle wave in which treatments were initiated. Furthermore, during the second experiment, the mean CL diameter was larger in letrozole-treated heifers, although there were no differences in plasma progesterone concentrations between treated and control animals. The effects on dominant follicle and CL diameter profiles appeared to be the result of the significantly increased plasma LH concentrations observed in letrozole-treated animals during both treatment approaches (single vs multiple dose). Incomplete and inconsistent inhibition of estradiol production and the lack of a surge on FSH observed in both experiments may be a result of insufficient circulating levels of letrozole during the treatment period. In summary, a single or multiple dose of letrozole did not induce regression of the extant dominant follicle, nor did it directly affect FSH release. Conversely, letrozole extended the lifespan of the dominant follicle, in association with increased endogenous levels of LH, thereby delaying the next FSH surge and subsequent follicular wave emergence. Results suggest that letrozole has potential as a non-steroidal method for controlling ovarian function in cattle, but further studies are warranted to clarify dosage and timing of treatment to predictably affect follicular wave dynamics in cattle.