Osteoporosis affects the human skeleton through the direct effects of the disease on the function and structure of bone. Individuals who are affected by osteoporosis may be subject to serious fractures and it is estimated that annually approximately 1.5 million fractures can be attributed to this disease [1]. The disease is categorized as the direct side effect of increased bone porosity and bone loss and is directly linked to estrogen deprivation [2]. Animal models are often used to make initial conclusions about the effects of the disease or pharmacological treatments. In this study, sheep were chosen as a representative animal model due to their similar metabolic characteristics to that of a human. Like most animals, the ovine does not undergo a natural menopause and an ovariectomy was necessary to replicate the condition. The study objective was to quantify compact bone density present in ovine at three months post ovariectomy. The study included 112 ovine separated into different treatment groups. The treatment groups were separated into 4 groups of 28 based on season of surgery: autumn, winter, spring, and summer. Each seasonal group was further divided into 2 groups of 14; the first group underwent an ovariectomy; and the second group underwent a sham surgery, in which the ovaries were visualized and handled but left in the abdomen. One group was sacrificed 3 months post operatively and the other group was sacrificed at 12 months post operatively. This study specifically looks at ewe sacrificed at 3 months. The radius from each sheep was cut into the anatomical sectors: cranial, caudal, craniolateral, craniomedial, caudomedial, and caudolateral. Each anatomical sector was turned into a microradiograph for analysis. Densitometry was performed to determine the density of each specimen using the estimated thickness of aluminum (ETA) as the key. Statistical analysis assessed the resulting data to understand the effects of treatment, season of sacrifice, season of surgery, and anatomical sector by comparing both mean ETA and standard deviation ETA to understand changes in bone density. The results revealed significant differences between the ovariectomy and sham groups as well as variation within season of surgery and season of sacrifice in both groups. Anatomical sector showed no significant variation. The differences in the thickness of aluminum seen in the sheep that underwent a sham operation can be attributed to the presence of estrogen. The sheep that underwent an ovariectomy showed differences in the estimated thickness of aluminum that can be attributed to other seasonal characteristics including the influence of Vitamin D. The results and conclusions within this study can be used to influence bone material characteristics and bone loss test protocols in future osteoporosis and estrogen depletion studies.

This study determined the effects of gestational obesity on the long-term metabolic status of the mother and if obesity management during or after pregnancy could attenuate these effects. At 120 days prior to estrus, 8 ewes received 100 percent of NRC nutrient requirements (control group) and 24 ewes had free access to feed (obesity induction). Beginning on day 42 of gestation, 8 obese ewes were restricted to 65 percent of NRC nutrient requirements. Following parturition, controls and all but one group of obese ewes were fed 100 percent of NRC nutrient requirements. At postpartum days (PPD) 1 and 150, glucose tolerance tests were administered to ewes. At both PPD1 and PPD150, obesity resulted in insulin resistance, impairment of whole-body glucose utilization, increased levels of circulating leptin, and altered profiles of amino acids in plasma; however, these effects were diminished in ewes receiving obesity management during or after gestation. Additionally at PPD150, obesity increased the circulating levels of ammonia and urea in ewes, which was prevented by realimentation to 100 percent NRC requirements. These results indicate that weight reduction in obese dams during pregnancy or after parturition can beneficially ameliorate the adverse effects of gestational obesity on the mother.

This thesis is a comparative study of Ewe and English based on the Grammatical categories of Determiners and Aspects. First, it deals with determiners with reference to sub-categories such as articles, demonstratives, pronouns and quantifiers. Second, it explores. the domain of aspects with particular attention to the differences between tense and aspect. The thesis also studies the grammatical categories of present, past and future with emphasis. on perfective and imperfective meanings. In this work, there is a general attempt to see how related the two languages are on structural and semantic levels.At the end of each part, the thesis specifies the contrasts between the two languages, makes an account of possible interfering areas, and analyses how they can help the instructor in teaching English to native Ewe students.

Thesis (Ph. D)--University of Pittsburgh, 1988. / Includes bibliographical references (p. 387-391).

A nutrição é o principal fator que interfere com o desempenho reprodutivo de mamíferos e vários metabólitos e hormônios, envolvidos no metabolismo energético, funcionam como sinalizadores para o eixo hipotálamo-hipófise-gonadal. O fato de a insulina ser o principal regulador da homeostase de glicose e exercer controle em diversas etapas do metabolismo de gorduras e proteínas, fez desse hormônio, ao longo do processo evolutivo, um modulador da reprodução. Neste estudo, no experimento 1, foi comparado o efeito da hiper e da hipoinsulinemia, no desempenho reprodutivo relacionado ao processo de superovulação em ovelhas. Para tanto foram utilizadas 27 ovelhas, distribuídas em 3 grupos: a) controle; b) grupo diabético (induzido pela aplicação I.V. de 50mg/kg de Alloxano); e c) grupo hiperinsulinêmico (suplementado com 1 UI/kg ao dia, S.C.). Todos os animais receberam um pessário vaginal, contendo 60 mg de medroxiprogesterona no D0 e foram superovulados, com 250UI de FSH em 6 aplicações, iniciadas no D10. No D12 aplicaram-se 250UI de eCG e 125 g de cloprostenol sódico. As ovelhas foram submetidas à monta natural e a colheita dos embriões foi realizada no sétimo dia após o início do estro. Em média, os teores de insulina medidos a partir da remoção dos pessários até a colheita dos embriões foram de 14,52±0,4 vs 10,18 ± 0,5 vs 20,05±0,9 UI/mL (P<0,01), respectivamente para os grupos controle, diabético e hiperinsulinêmico. Os valores para glicemia, medida no mesmo período, para os grupos controle, diabético e hiperinsulinêmico foram de 83,1±2,1 mg/dL vs 241,2±9,2 mg/dL vs 53,9±2,7 mg/dL (P<0,01), respectivamente. O grupo diabético apresentou menor produção de corpos lúteos que os animais controle e hiperinsulinêmicos (5 ±1,1 vs 10.3±1,9 e 11,3±1, P<0,01); pior qualidade do CL (IQCL de 2,3±0,3 vs 1.6±0,1 e 1.3±0,1, P<0,01), menor número de embriões (2.3±1.2 vs 7.9±1.97e 7.4±1.2, P<0,01), que por sua vez também foram de pior qualidade (IQE de 2.9±0.2 vs 2±0.1 e 1.7±0.1, P<0,01). De um modo geral, os animais hiperinsulinêmicos apresentaram desempenho reprodutivo semelhante aos do grupo controle; entretanto, embora o número de embriões colhidos não tenha sido estatísticamente diferente, os dados são sugestivos de que o estado de hiperinsulinemia pode favorecer o crescimento embrionário, acelerando seu desenvolvimento. Histologicamente, os CL do grupo diabético se apresentaram com hipotrofia das LLC, aumento no número de células apoptóticas por campo, quando comparados aos dos tratamentos controle e hiperinsulinêmico. Adicionalmente, no experimento 2, foram testadas formulações de nanopartículas de quitosana, para liberação sustentada de insulina, bem como permeação da mucosa gastrintestinal. A formulação de insulina nanoestruturada, sem proteção lipídica,administrada pela via SC, liberou 92,1 ± 3,01% da quantidade inicial de insulina, in vitro, porém o padrão desejado de liberação sustentada não foi atingido. No teste in vivo, a redução da glicemia foi apenas parcial (em média 60,8±3,2% em relação à linha de base). O sistema composto por nanopartículas incorporadas à matriz lipídica, no teste in vitro, liberou apenas 15,6 ± 4,9% da quantidade inicial. Entretanto, quando administrada pela via oral, no teste in vivo, reduziu, embora parcialmente, a glicemia de ovinos diabéticos alloxano induzidos (em média 79,88±4,3% em relação à linha de base). Concluiu-se, com base no experimento 1 que, na dose empregada, a insulina não foi capaz de produzir benefícios reprodutivos que justifiquem seu uso em protocolos de superovulação de ovelhas. As concentrações subfisiológicas de insulina, observadas nos animais diabéticos podem ser responsáveis por uma série de alterações metabólicas, que, em conjunto, comprometeram os índices de desempenho reprodutivo, relacionados ao processo de superovulação e induziram um quadro inicial de regressão de CL. Com isso, observou-se que o uso de ovelhas, como modelo animal, para estudo dos efeitos reprodutivos da insulina, foi satisfatório. Pela análise do experimento 2, concluiu-se que o sistema de nanopartículas revestidas por lipídios foi capaz de carrear a insulina, ao longo do trato digestivo de um ruminante, no teste in vivo, e compatibilizar sua permeação através da mucosa intestinal, mantendo a atividade biológica do hormônio, o que consiste em um fato inédito. / Nutrition is the main factor that interferes with the reproductive development of all mammals and many of the matobolites and hormones involved in the energetic metabolism work as signaling factors for the hypothalamic-pituitary-gonadal axis. The fact of insulin being the main regulator of glucose homeostasis and having control in various steps of fat and protein metabolism, has made this hormone, along the evolution process, a reproductive modulator. In the first experiment of this study, the effect of hyper and hypoinsulinemia were compared as to how it affects the reproductive performance related to superovulation in sheep. For this, 27 sheep were used, distributed in 3 groups a) control, b) diabetic group (induced by IV injection of 50mg/kg of Alloxan); and c) hyperinsulinemic group (supplemented with 1 UI/kg per day, SC). All animals received a vaginal pessary, containing 60 mg of medroxiprogesterone on D0 and were superovulated, with 250UI of FSH in 6 applications, starting on D10. On D12 250UI of eCG and 125 g of sodium cloprostenol were administered. The sheep were submitted to natural breeding and embryo collection was performed on the seventh day after the beginning of estrus. In average, the insulin levels recorded starting on the day of pessary removal until the day of embryo collection were 14,52±0,4 vs 10,18 ± 0,5 vs 20,05±0,9 UI/mL (P<0,01), respectively for the control, diabetic and hyperinsulinemic groups. The values for glycemia, measured during the same period, for the control, diabetic and hyperinsulinemic groups were 83,1±2,1 mg/dL vs 241,2±9,2 mg/dL vs 53,9±2,7 mg/dL (P<0,01), respectively. The diabetic group showed less corpus lutea production than the control and hyperinsulinemic groups (5 ±1,1 vs 10.3±1,9 and 11,3±1, P<0,01); worse CL quality (IQCL de 2,3±0,3 vs 1.6±0,1 and 1.3±0,1, P<0,01), less number of embryos (2.3±1.2 vs 7.9±1.97 and 7.4±1.2, P<0,01), which by its turn were also of worse quality (IQE de 2.9±0.2 vs 2±0.1 and 1.7±0.1, P<0,01). Overall, hyperinsulinemic animals presented a reproductive performance similar to the control group; however, although the number of embryos recovered were not statiscally different, the data suggest that the state of hyperinsulinemy can favor embryo growth, acceleratting its development. Histologically, the CLs from the diabetic group showed hypotrophy of LLC and an increase in the number of apoptotic cells per field when compared to the control and hyperinsulinemic treatments. In addiction, the second experiment, chitosan nanoparticles formulations were tested, for sustained release of insulin as well as gastrointestinal mucosal permeability. The nanostructured insulin formulation without lipid protection, administered SC, released 92,1 ± 3,01% of the initial insulin amount, in vitro, however, the desired standard for sustained release was not reached. In the in vivo test, the reduction in glycemia was only partial (on average 60,8±3,2% in relation to base line ). In the in vitro test, the system made up of nanoparticles incorporated to the lipid matrix, released only 15,6 ± 4,9% of the initial amount. However, when administered orally, in the in vivo test, it reduced, although only partially, the glycemia of the alloxan induced diabetic ovines (on average 79,88±4,3% in relation to base line). In conclusion, based on the first experiment, the applied insulin dose was not able to produce any reproductive benefits that may justify its use in sheep superovulation protocols. The sub-physiologic insulin concentrations observed in the diabetic animals may be responsable for various metabolic alterations, that together compromised the reproductive performance levels related to the superovulation process and induced an initial state of CL regression. With that, it was noticed that the use of sheep as an animal model for the study of the effects of insulin on reproduction was satisfatory. By analyzing the second experiment, it was concluded that the nanoparticles system coated with lipids was able to carry insulin along the ruminant digestive during the in vivo test, show permeability through the intestinal mucosa, maintaining the hormone biologic activity, which is a new and unpublished fact.

Body size was estimated by multiplying the average of the hip and chest widths by body length and chest depth in 208 ewes of three genotypes: Targhee x Targhee (TXT), Finn x Targhee (FXT), and Suffolk x Targhee (SXT), and two seasons, fall (pre-breeding) and spring (post-lambing). The estimated ewe body size did not fluctuate with season, body condition or physiological stress and proved to be a constant measure of size. Measuring linear body dimension is time consuming; and hence ewe body size can best be approximated by the chest depth in a regression equation. In the absence of linear measurements, the ewe body weight remained to be the best measure of size. In evaluating relationships, there was a tendency for ewe body size and weight to be positively (P<.05) related to reproductive traits and body condition to be negatively (P<.05) related. The heavier ewes were more prolific in the FXT ewes and the lighter ewes in the SXT ewes, the TXT ewes being intermediate. Lamb production on per unit size or weight bases tended to be negatively (P<.05) related with weight and/or size in all the three genotypes. This relationship was stronger in the SXT ewes than in the TXT ewes, which in turn was stronger than in the FXT ewes, suggesting that the heavier/larger the ewe the less she produced in relation to her weight or size.

The purpose of this study is to characterize the compact bone remodeling of the ovariectomized ewe and its effectiveness as an animal model for studies of postmenopausal osteoporosis. The use of animal models is a beneficial way to evaluate the effectiveness of medical devices and therapeutic agents for treating diseases. The ovine model is an attractive option because of its large size and similar bone remodeling to humans. However, this species, like most animals, does not undergo a natural menopause, making an ovariectomy necessary for studies of estrogen depletion. The OVX (ovariectomized) sheep has been widely accepted as a model for loss of BMD. However, even with its advantages seasonality has been shown to have an effect on the bone remodeling in the ovine model. Thus, this study focused on characterizing the seasonal and anatomical variation in the compact bone remodeling in the ovine model. In doing so, 28 skeletally mature Columbia-Rambouillet cross ewes underwent an ovariectomy. The animals were divided into groups of 7 based on each season: autumn, winter, spring, and summer. During its specified season each group of 7 underwent ovariectomy and then was sacrificed 12 months post-surgery. The radii and ulnae were harvested and then divided into 6 anatomical locations: craniolateral, cranial, craniomedial, craniolateral, caudal, and caudomedial. This allowed for analysis through the fabrication of microradiographs. Histomorphometric analysis involved measurements to quantify the bone volume to tissue volume ratio, the percent of tissue and material remodeling, the mean secondary osteonal radius, and number of tissue and material cement line interfaces. Densitometry analysis was then performed to determine the density of each sector relative to an aluminum step wedge, serving as the key. Using a 2-way repeated measures ANOVA statistical analysis was performed to evaluate for seasonal and anatomical differences as well as a possible interaction between season and anatomical region. The OVX data showed significant seasonal and anatomical differences as well as seasonality within anatomical section in the remodeling parameters. When this data was combined with earlier data from a control, sham-cohort from the same experiment a 3-way ANOVA was also performed to evaluate the effects of season, anatomical sector, and treatment on the remodeling parameters. Again, there was significant seasonal, anatomical, and treatment differences, as well as interactions of all three. The results of this study showed that varying levels of remodeling occurs in the adult OVX ovine model and differences between the control and OVX model can be attributed to seasonal and anatomical variations. Thus, it will be an important consideration when developing new test protocols for research incorporating the sheep model for studying osteoporosis.

The objectives of this dissertation were to evaluate genetic influences on ewe productivity, its growth and reproductive components, and measures indicative of accelerated lambing performance. Genetic parameters were estimated for total weight of litter weaned per ewe lambing (TW) and its components, number of lambs born (NB), number of lambs weaned (NW) and average weight of lambs weaned (AW), measured as traits of the ewe, and lamb survival (LS) and weaning weight (WW), measured as traits of the lamb, in Katahdin sheep. Heritabilities of TW, NB, NW, and AW, were 0.12, 0.12, 0.09, and 0.13, respectively. Heritability of WW was 0.15 to 0.20. Genetic effects on LS were negligible. Genetic correlation of TW with NB, NW, and AW averaged 0.30, 0.90, and 0.74, respectively, those of NB with NW and AW averaged 0.72 and 0.01, respectively, and that between NW and AW averaged 0.50. Direct genetic effects on WW were independent of NB and NW, but correlation between maternal genetic effects on WW and animal genetic effects on NW averaged 0.35. Ewe fertility, NB, LS, and WW were modeled using stochastic simulation and used to derive NW, AW, and TW to test alternative predictors of genetic merit for TW. A random 8% of WW observations were set to missing values and AW and TW were recalculated to evaluate the effects of data reporting inconsistencies on efficacy of different prediction strategies. Four alternative predictors of estimated breeding values (EBV) for TW involved direct univariate prediction (TW1), an index of EBV for NW and AW (TW2), indirect prediction using data for NW and AW and genetic correlations among NW, AW and TW (TW3), and indirect prediction augmenting TW3 with data and genetic correlations involving NB (TW4). To validate efficacy of predictors, daughter data sets were generated from the original ewes and their realized TW were regressed on alternative predictors. Regression coefficients from TW1, TW3, and TW4 were close to the expected value of 0.50 whereas those from TW2 were less than 0.50. Model Rsquare statistics were similar among predictors when there were no missing WW data but regressions involving TW1 had lowest model R-square when some WW data was missing. Ewe lamb fertility (ELF), ages at first, second, and third lambings (AGE1 to AGE3), first and second lambing intervals (INT1 and INT2), and number of lambings by 38 mo of age (LAMB3) were evaluated for an accelerated lambing Polypay flock. Relationships among these traits and NB and WW were estimated. Heritability of ELF, AGE1, AGE2, AGE3, INT1, INT2, and LAMB3 were 0.14, 0.39, 0.28, 0.36, 0.00, 0.09, and 0.27, respectively. Heritability of AGE2 and AGE3 were negligible after accounting for variation in AGE1. Genetic correlations of ELF with AGE1 and AGE2 were -0.89, -0.91, respectively, and that with LAMB3 was 0.89. Genetic correlations of LAMB3 with AGE1 and AGE2 were -0.49 and -1.00, respectively. Genetic correlations of ELF and LAMB3 with direct genetic effects on WW were close to -0.70, but correlations with maternal genetic effects on WW were 0.88 and 0.58, respectively. Prolificacy was independent of ELF and LAMB3. / Ph. D.

Prostaglandin (PG) F₂α, is luteolytic in ewes, but the exact mechanism for this effect is not clear. 6α-Methyl-17α hydroxyprogesterone acetate (MPA) reduces uterine secretion of PGF₂α, but it does not interfere with luteolysis. The mechanism by which MPA suppresses PGF₂α and permits luteal regression remains to be determined. Three experiments were conducted to determine whether MPA reduces PGF₂α in uteroovarian blood, causes the release of PGF₂α before d 14 of the estrous cycle, or increases sensitivity of corpora lutea (CL) to exogenous PGF₂α. In Exp. 1 and 2, blank (control) or MPA-impregnated pessaries were inserted six days after estrus (i.e., d 6). Jugular, vena caval, and(or) uteroovarian blood samples were collected frequently (i.e., at .5 h intervals during twice daily collection periods of 2 h each) before and after luteolysis. The MPA reduced (P < .05) jugular, vena caval, and uteroovarian concentrations of PGF₂α, but MPA did not affect the interval from estrus to the first day of luteolysis. Average PGF₂α concentrations in uteroovarian and vena caval blood differed (P < .05) but the profiles did not. In Exp. 3, blank (control), MPA-impregnated, or no (untreated) pessaries were inserted on d 7. A minimal luteolytic dose of PGF₂α (Lutalyse ®; 4 mg/58 kg of body weight) was administered on d 8 or 12, blood samples were collected and the CL were collected 48 h after PGF₂α treatment. The MPA increased the sensitivity of CL to exogenous PGF₂α. Progesterone and CL weights were less (P < .05) in MPA-treated ewes than in control and untreated ewes given PGF₂α on d-8. The MPA-treated ewes given PGF₂α on d-12 had smaller CL than did ewes in the other two groups, but MPA did not affect progesterone concentrations. In summary, MPA reduces the ability of the uterus to secrete PGF 7a and increases the sensitivity of CL to a minimal luteolytic dose of PGF₂α. This information should improve our ability to control the estrous cycle ewes. / Master of Science

Twenty-three crossbred ewes were utilized during 1995, to investigate the possibility of xenogenous capacitation, using stallion spermatozoa. Ewes were grouped according to endocrine status as anestrus (n = 12) or estrus (n = 11) and were surgically inseminated with either fresh (FR), cooled/extended (FCE) or frozen/thawed (FZ) spermatozoal samples. The capacitation response of spermatozoa recovered from oviductal flushing 4 - 6 hours post-insemination, was assessed using a chlortetracycline (CTC) fluorescent probe. Data were recorded as percentages of spermatozoa exhibiting CTC staining patterns characteristic of capacitated (PCAP), unreacted (PUR) and acrosome reacted (PAR) sperm. Mean PCAP was not significantly different in estrous ewes despite an increasing trend. Mean PUR and PAR were also not different. Differences in capacitated, acrosome reacted and unreacted spermatozoa in inseminated and recovered samples (CAPDIF, ARDIF and URDIF, respectively) were analyzed. CAPDIF was significantly different between treatment groups (p < 0.01). CAPDIF was also significantly greater for the estrus versus anestrus group (p < 0.05). Total number of spermatozoa recovered (RTOTAL) was recorded. More spermatozoa were recovered from estrus ewes, however significance was not established. Mean number of spermatozoa recovered was 37.9 ± 35.9 per ewe. Treatment significantly affected RTOTAL in estrus animals (p = 0.01). FR samples had the highest recovery. Results suggest that xenogenous capacitation of stallion semen may occur in the reproductive tract of the ewe. Implications of these results are discussed in regards to their application of xenogenous gamete intrafallopian transfer (X-GIFT) as a treatment option for certain infertility problems in the mare. / Master of Science