Thesis (Ph. D.)--University of Washington, 1997. / Vita. Includes bibliographical references (leaves [96]-100).

Universiẗat, Diss., 2003--München.

Universiẗat, Diss., 2004--München.

Background: Insulin is an important hormone for glucose homeostasis. It is released from β-cells in the endocrine pancreas as a response to increased concentrations of plasma glucose. The major effect of insulin is the facilitation of cellular uptake and storage of glucose as glycogen. Insulinomas are tumours that produce excessive amounts of insulin resulting in hypoglycaemia. The condition has been observed in dogs and cats and is often malignant. One part of establishing the diagnosis is confirmation of elevated concentrations of insulin in a hypoglycaemic sample. Aim: The aim of this study was to evaluate if an equine-optimized insulin ELISA (Mercodia AB, Uppsala, Sweden) is useful for analysis of insulin in canine and feline serum samples when insulinoma is suspected. Material and methods: All samples were analysed with Equine insulin ELISA. Precision, linearity and effects of haemolysis were studied. The stability of insulin was evaluated after storage in 4°C, room temperature and after repeated freezing and thawing. A reference interval was constructed for both canine and feline samples. Results: Total precision expressed as CV was 4.4 – 18.9 %. The method was linear up to at least 100 mU/L for dogs and 15 mU/L for cats. Reference interval for cats was <11.6 mU/L, due to few healthy animals no reference interval for dogs could be established. Stability was acceptable for up to four days. No effects of haemolysis were detected. Conclusion: Mercodia Equine insulin ELISA is suitable for analysis of insulin in serum from dogs and cats when suspecting insulinoma.

Primary aldosteronism (PA) is a common form of secondary hypertension with an international prevalence rates between 5 and 10 %. It is characterized by a high autonomous aldosterone production that causes cardiovascular damage, renin suppression, hypertension, sodium retention, potassium excretion and hypokalemia. The screening of PA is a simple test measuring aldosterone to renin ratio (ARR) with immunoassay method. This test is currently considered as the most reliable screening tool for PA.     The main objective of the study was to evaluate an ELISA-method, for detection of aldosterone and renin in blood plasma, to be used for routine analysis in the laboratory. The second aim was to investigate the effect of refreezing samples, considering that cryoactivation of prorenin might occur.     One hundred blood samples were analysed, in regard to aldosterone and renin, by using two commercial ELISA assays (DRG ELISA from DRG Diagnostics, Germany) on a Dynex DS2 instrument. In addition, the accuracy and precision of the methods were calculated. The effect of refreezing was investigated with a series of eight samples, which were analyzed twice on the same instrument.     Both assays performed well. The resulting data showed good precision and accuracy. The correlation between the original and refreezed samples was good, r = 0.989 and r = 1.0 for aldosterone and renin respectively. Considering that the study only included eight samples, further investigation is recommended.     Evaluation showed that both immunoassays are reliable in diagnostic use and the ELISA-method is suitable to implement in the laboratory for routine analysis.

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Orientador: José Maurício Barbanti Duarte / Banca: Paulo Henrique Franceschini / Banca: Claudio Alvarenga de Oliveira / Resumo: Atualmente, o conhecimento sobre a fisiologia e a biologia reprodutiva de cervos-do-pantanal (Blastocerus dichotomus) é escasso e as lacunas existentes aumentam a vulnerabilidade da espécie à extinção. Neste contexto, os objetivos deste trabalho foram: (1) caracterizar alguns parâmetros relacionados à fisiologia reprodutiva de fêmeas de cervos-do-pantanal e (2) fornecer métodos adequados para o acesso às diferentes fases do ciclo reprodutivo. Para isso, seis fêmeas da espécie foram utilizadas para a caracterização hormonal do ciclo estral, da gestação e do período pós-parto. Amostras fecais foram colhidas diariamente para o monitoramento do ciclo estral, duas vezes por semana durante a gestação e em dias alternados no período pós-parto. As dosagens das progestinas fecais foram feitas por EIA. Foram monitorados 16 ciclos estrais completos, cuja duração média foi de 21,3±1,3 dias (6,4±1,2d fase inter-luteal e 14,8±1,3d fase luteal). As concentrações médias de progestinas fecais para as fases inter-luteal e luteal do ciclo estral foram 834±311ng g-1 e 3979±1611ng g-1, respectivamente, e o valor limite entre elas foi de 1457ng g-1. Quatro fêmeas levaram a gestação a termo, determinando um período de gestação médio de 253±4 dias. Houve um aumento significativo (p<0,05) na concentração de progestinas fecais a partir do quarto mês de gestação, o que pôde direcionar o diagnóstico presuntivo de gestação na espécie. Assim, concentrações de progestinas fecais 15250ng g-1 foram indicativas de gestação. No período de 60 dias pós-parto em que as fêmeas foram monitoradas, 75% (3/4) delas apresentaram estro e/ou retomaram a atividade ovariana. / Abstract: The scarce knowledge about reproductive physiology and biology in marsh deer (Blastocerus dichotomus) increases the vulnerability of this species to the extinction. Thus, the aims of this study were: (1) to characterize some parameters related to the reproductive physiology of females marsh deer and (2) to turn available adequate tools for the assessment of the reproductive cycle's phases. Six females and one male were used to the hormonal characterization of the estrous cycle, pregnancy and post parturition period. Fecal samples were collected daily during the estrous cycle, twice a week during the pregnancy and in alternated days during the post parturition period of monitoring and the amount of fecal progestins was evaluated by enzyme immunoassay. The mean length of the estrous cycle was 21.3±1.3 days (6.4±1.2d interluteal and 14.8±1.3d luteal phase) and the fecal progestins concentrations were 834±311ng g-1 and 3979±1611ng g-1, respectively for the inter-luteal and luteal phase. The limit value between these phases was 1457ng g-1. Four females became pregnant and exposed a mean pregnancy period of 253±4 days. There was a significant increase (p<0,0001) in the fecal progestins excretion since the fourth month of pregnancy, that could provide guidelines for the pregnancy diagnosis in the species. Thus, fecal progestins concentrations 15250ng g-1 were indicative of pregnancy. Seventy five percent (3/4) of the females exhibit estrous and/or restarted the ovarian function during the post parturition period of monitoring. / Mestre