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PadronizaÃÃo de modelo de ligadura da artÃria uterina em ratas nÃo-grÃviadas, seus efeitos sobre a isquemia uterina direta e suas repercussÃes reprodutivas. / Standardized model of uterine artery ligation in non-pregnant rats, their effects on uterine ischemia and its direct impact on reproductiveJosà Ananias Vasconcelos Neto 02 December 2009 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A obstruÃÃo das artÃrias uterinas promove isquemia e/ou necrose no Ãtero, no entanto nÃo se conhece com que intensidade essas lesÃes ocorrem. Os objetivos deste estudo sÃo: descrever e padronizar uma tÃcnica de ligadura da artÃria uterina (LAU) direita em ratas nÃo grÃvidas e avaliar os efeitos deste modelo em Ãteros e ovÃrios de ratas. Estudo experimental, utilizando 64 ratos, 48 fÃmeas e 16 machos (Rattus norvergicus, variedade albina) maduros, de fertilidade comprovada. As ratas foram alocadas aleatoriamente em 8 grupos, de seis indivÃduos. Quatro grupos foram submetidos à tÃcnica de LAU direita e sacrificados nos dias 1, 7, 14 e 21 apÃs o procedimento. Outros 3 grupos foram acasalados nos dias 1, 7 e 14 apÃs a LAU, e comparados com o grupo controle quanto à fertilidade. ApÃs o sacrifÃcio, eram retirados, para anÃlise histopatolÃgica, os ovÃrios, e os hemi-Ãteros. Realizou-se anÃlise histolÃgica, que avaliou o Ãtero quanto à congestÃo, hemorragia, edema intersticial e perda de coesÃo celular atravÃs de um escore que varia de 0 a 3. Os ovÃrios foram avaliados de acordo com o nÃmero de folÃculos em desenvolvimento e corpos lÃteos. Para a anÃlise estatÃstica foi utilizado o software SPSS (Statistical Package for Social Sciences) versÃo 13.0, p < 0,05 foi considerado estatisticamente significativo. O projeto de pesquisa foi enviado para a ComissÃo de Ãtica em Pesquisa Animal (CEPA) da Universidade Federal do CearÃ, com protocolo de nÃmero 90/08. O modelo da tÃcnica foi realizado em ratas nÃo-grÃvidas utilizando a ligadura na porÃÃo inferior da artÃria uterina direita, mantendo o hemi-Ãtero esquerdo (HUE) como controle. Em nenhum momento dos dias de sacrifÃcio, apÃs a LAU (1, 7, 14 e 21 dias), houve diferenÃa entre os escores histolÃgicos de isquemia dos hemi-Ãteros direitos (HUD) distais e proximais (p > 0,05). Os escores histolÃgicos de isquemia dos hemi-Ãteros direitos no decorrer do tempo aumentaram substancialmente a partir do 7 dia (p=0,003). Da mesma maneira ocorreu com os hemi-Ãteros esquerdos (p=0,001). A Ãnica diferenÃa significativa observada na comparaÃÃo dos escores dos hemi-Ãteros direito e esquerdo ocorreu no 1 dia apÃs LAU (p=0,026), à custa de congestÃo. Os ovÃrios esquerdos nÃo apresentaram alteraÃÃes no nÃmero de folÃculos e de corpos lÃteos apÃs LAU e, os ovÃrios direitos apresentaram nÃmero de folÃculos e corpos lÃteos semelhante ao controle a partir do 21 dia da LAU. A percentagem de ratas grÃvidas que foram submetidas a LAU foi de 44,4%, comparado com 100% das ratas controle que engravidaram (p=0,024). Observou-se ainda uma reduÃÃo na mÃdia do nÃmero de fetos por rata (p=0,029). Pode-se concluir que o modelo estabelecido foi efetivo e de fÃcil reprodutibilidade, bem como as alteraÃÃes histolÃgicas encontradas no Ãtero ocorrem de forma discreta. Em relaÃÃo a funÃÃo ovulatÃria, pode-se dizer que o nÃmero de folÃculos e corpos lÃteos dos ovÃrios direitos, a partir do 21 dia, permaneceram semelhantes aos dos esquerdos (controle). A fertilidade, porÃm, mostrou-se reduzida apÃs o estabelecimento desta tÃcnica. / The obstruction of the uterine arteries promotes ischemia or necrosis in the uterus. However it is not known how strongly these injuries occur. The objectives of this study are to describe and standardize a technique of right uterine artery ligation (UAL) in non-pregnant rats and to evaluate the effects of this model in the uteri and ovaries of female rats. An experimental study using 64 rats, 48 females and 16 males (Rattus norvegicus, albino variety) mature, with proven fertility. The rats were randomly allocated into 8 groups of six individuals. Four groups were subjected to the technique of right UAL and sacrificed on days 1, 7, 14 and 21 after the procedure. Other 3 groups were mated on days 1, 7 and 14 after the UAL, and compared with the control group regarding fertility. After sacrifice, ovaries and the hemi-uteri, were removed for histological analysis wich was carried out histological analysis, which evaluated the uteri and the congestion, hemorrhage, interstitial edema and loss of cell cohesion through a score ranging from 0 to 3. Ovaries were evaluated according to the number of follicles and corpora lutea. For statistical analysis we used SPSS software (Statistical Package for Social Sciences) version 13.0, p <0.05 was considered statistically significant. The research project was submitted to the Ethics Committee on Animal Research (ECAR) Federal University of CearÃ, under the number 90/08. The model of the technique was performed in non-pregnant rats using ligation in the lower portion of the right uterine artery, keeping the left hemi-uteri (LHU) as control. At no time during periods of ischemia established (1, 7, 14 and 21 days) there was any difference between the histological scores of ischemia of the right hemi-uteri (RHU) distal and proximal (p> 0.05). The histological scores of ischemia of the right hemi-uteri over time increased significantly from day 7 (p = 0.003), just as occurred with the left hemi-uteri (p = 0.001). The only significant difference observed when comparing the scores of the right hemi-uteri and left occurred on day 1 after UAL (p = 0.026), due to congestion. The left ovary showed no changes in the number of follicles and corpora lutea after UAL, and the right ovary showed a number of follicles and corpora lutea similar to control from the 21th day of UAL. The percentage of pregnant rats that were subjected to ischemia was 44.4%, compared with 100% of control rats that became pregnant (p = 0.024). There was also a reduction in the average number of fetuses per mother (p = 0.029). It can be concluded that the model established was effective and highly reproducible, and histological changes found in the uteri are mild. As to ovulatory function, one can say that the number of follicles and corpora lutea of the right ovaries, after 21 days, remained similar to the left (control). Fertility, however, was reduced after the establishment of this technique.
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Padronização do modelo experimental de lesão da medula espinal e avaliação da lesão neurológica em camundongos / Standardization of a spinal cord lesion model and neurologic evaluation using micePaulo Alvim Borges 16 January 2018 (has links)
INTRODUÇÃO: A lesão da medula espinal é um dos grandes desafios da medicina. Apesar de décadas de pesquisa sobre o assunto, seu tratamento ainda não é satisfatório. A padronização de modelos de lesão da medula espinal permite a reprodutibilidade e a análise dos resultados sendo importante para a pesquisa sobre o tema. OBJETIVO: Validar a padronização de um modelo de lesão da medula espinal e avaliação da lesão neurológica em camundongos. MÉTODOS: Submetemos 30 camundongos BalbC divididos em 4 grupos experimentais e um grupo controle à lesão da medula espinal torácica por queda de peso de diferentes alturas (gerando lesões de graus variados). O grupo controle (SHAM) foi submetido apenas à laminectomia. Os camundongos foram avaliados por seis semanas durante as quais foram aplicadas escalas de avaliação funcional motora. Após seis semanas os animais foram sacrificados para avaliação histológica das medulas espinais lesadas. Os achados foram correlacionados entre si para validar se a lesão foi efetiva e se os grupos diferenciaram-se entre os diferentes graus de lesão. Adicionalmente avaliamos se as escalas utilizadas são aplicáveis e se são fiéis aos achados histológicos. RESULTADOS: Seis dos trinta camundongos do experimentos evoluíram para óbito sendo um do Grupo 3, um do Grupo 4 e quatro do Grupo 5. Um camundongo do Grupo 4 apresentou autofagia. O Grupo 5 foi excluído do experimento por alta mortalidade e perda de dados. Todas as escalas funcionais estudadas foram estatisticamente diferentes entre si e demonstraram evolução durante o experimento. Os achados foram confirmados por histologia e apresentaram uma correlação forte com as escalas BBB e BMS e moderada a forte com a escala MFS. A Escada Horizontal apresentou forte correlação com a degeneração neurológica porém não apresentou correlação com os demais parâmetros histológicos estudados. CONCLUSÃO: O modelo de lesão da medula espinal em camundongos apresentado neste estudo é efetivo, confiável e reprodutível, com exceção da lesão causada por queda de peso (10g) de 50mm de altura, que traz mortalidade inaceitável. Das escalas estudadas, BBB e BMS são as mais confiáveis, enquanto que a Escada Horizontal tem seu uso discutível / INTRODUCTION: Spinal cord lesion is a great medical challenge. Even with many decades of research, no satisfactory treatment is available yet. The standardization of animal experimentation models makes the spinal cord lesion reproducible allowing a reliable analysis of the results. Hence, standardization is a major concern in spinal cord lesion research. OBJECTIVE: To validate the standardization of a spinal cord lesion model with neurologic evaluation using mice. METHODS: Thirty BalbC mice were divided in four experimental groups and one control group and submitted to spinal cord lesion produced by weight drop from different heights (producing different severity lesions). The control group (SHAM) was submitted to laminectomy only. Every mice was followed up for six weeks during which functional motor scales were applied. After six weeks the animals were sacrificed for histological examination. Findings were correlat-ed to confirm if the spinal cord lesion was effective and if the groups were dif-ferent between themselves. Additionally all functional motor scales were corre-lated with the histological findings to confirm if the scales are reliable and truly represented the spinal cord lesion. RESULTS: Six mice died during the experi-mentation period (one mouse from the Group 3, one mouse from the Group 4 and four mice from Group 5). One mouse from Group 4 presented autophagia and was excluded from the experiment. Group 5 was excluded from the exper-iment for high mortality rates and data loss. All functional motor scales applied demonstrated significant results with moderate or strong correlation with the histological findings. The Horizontal Ladder scale had strong correlation with neurologic degeneration but had weak or worse correlation with the rest of the histological parameters studied. CONCLUSION: The spinal cord lesion model using mice presented in this study is reliable and reproducible, excluding the lesion produced by a weight drop (10g) from 50mm, which brings unacceptable mortality rate. Of all fuctional motor scales studied, BBB and BMS scales are the most reliable. The use of the Horizontal Ladder scale, however, must be carefully evaluated
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Characterization of a mouse model of shrimp allergy.January 2007 (has links)
Lee, Yuen Shan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 81-102). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / Table of contents --- p.vi / List of Figures --- p.ix / List of Abbreviations --- p.xi / Chapter Chapter 1. --- General introduction --- p.1 / Chapter Chapter 2. --- Literature review / Chapter 2.1 --- History of food allergy research --- p.3 / Chapter 2.2 --- Prevalence of food allergy --- p.4 / Chapter 2.3 --- Clinical symptoms of food allergy --- p.6 / Chapter 2.4 --- Mechanism of food allergy --- p.6 / Chapter 2.4.1 --- Properties of food allergens --- p.7 / Chapter 2.4.2 --- Exposures to food allergens in the gastrointestinal tract --- p.8 / Chapter 2.4.3 --- Oral tolerance and its relationship to food allergy --- p.9 / Chapter 2.4.4 --- Cellular mechanism of food allergy --- p.13 / Chapter 2.5 --- Studies on seafood allergies and allergens --- p.17 / Chapter 2.6 --- Use of animal models in the study of food allergy --- p.22 / Chapter 2.6.1 --- Selection of species and strain for developing animal models --- p.22 / Chapter 2.6.2 --- Parameters of sensitization protocol --- p.25 / Chapter 2.6.3 --- Lessons from animal models --- p.27 / Chapter 2.6.3.1 --- Investigations on pathogenesis of food allergy --- p.27 / Chapter 2.6.3.2 --- Studies on development of therapeutic strategies --- p.28 / Chapter Chapter 3. --- Characterization of hypersensitive responses to recombinant shrimp tropomyosin in mice / Chapter 3.1 --- Introduction --- p.30 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Preparation of the recombinant shrimp tropomyosin / Chapter 3.2.1.1 --- Expression of the recombinant shrimp tropomyosin --- p.32 / Chapter 3.2.1.2 --- Extraction and purification of the recombinant protein under native condition --- p.32 / Chapter 3.2.1.3 --- Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.33 / Chapter 3.2.1.4 --- Quantification of the recombinant protein and detection of level of endotoxin in the protein --- p.34 / Chapter 3.2.2 --- Characterization of hypersensitive responsesin mice / Chapter 3.2.2.1 --- Mice --- p.37 / Chapter 3.2.2.2 --- Sensitization and challenge of mice --- p.37 / Chapter 3.2.2.3 --- Assessment of systemic anaphylaxis responses --- p.38 / Chapter 3.2.2.4 --- Detection of shrimp tropomyosin specific IgE level --- p.39 / Chapter 3.2.2.5 --- Passive cutaneous anaphylaxis (PCA) test --- p.40 / Chapter 3.2.2.6 --- In vitro proliferation assay under stimulation of shrimp tropomyosin --- p.40 / Chapter 3.2.2.7 --- Cytokine profile of splenocytes --- p.42 / Chapter 3.2.2.8 --- Histological examination of small intestine --- p.44 / Chapter 3.2.2.9 --- Statistical analysis --- p.45 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Preparation of the recombinant shrimp tropomyosin --- p.47 / Chapter 3.3.2 --- Induction of systemic anaphylaxis responses after challenge --- p.48 / Chapter 3.3.3 --- Elevated level of shrimp tropomyosin specific IgE --- p.49 / Chapter 3.3.4 --- Passive cutaneous anaphylaxis (PCA) reactions --- p.50 / Chapter 3.3.5 --- Proliferation response of splenocytes under in vitro stimulation --- p.54 / Chapter 3.3.6 --- Cytokine profiles of restimulated splenocytes --- p.58 / Chapter 3.3.7 --- Histology of small intestine --- p.65 / Chapter 3.4 --- Discussion --- p.68 / Chapter Chapter 4. --- General conclusion --- p.78 / References --- p.81
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Efeitos da exposição neonatal a esteróides sexuais no hipotálamo e ovário de ratas adultas: modelos animais da síndrome dos ovários policísticos / Effects of neonatal exposition to sex steroids on hypothalamus and ovary of adult female rats: animal models of polycystic ovary syndromeMarcondes, Rodrigo Rodrigues 13 December 2012 (has links)
A síndrome dos ovários policísticos (SOP) é o distúrbio endócrino mais frequente em mulheres em idade reprodutiva. Sua etiologia é desconhecida, mas recentemente, fatores ambientais, como o excesso esteróides sexuais em fases precoces da vida, têm sido implicados na origem da SOP. Em ratas, o excesso de androgênios ou estrogênios na vida neonatal altera a função do eixo hipotálamo-hipófise-gonadal e induz alterações reprodutivas e metabólicas similares às observadas na SOP em humanos. O objetivo deste estudo foi analisar a expressão dos genes relacionados ao controle da secreção de GnRH (Gnrh, Gnrhr, Kiss1, Kiss1r e Ar) no hipotálamo de modelos animais de SOP e avaliar a correlação com a morfologia ovariana, níveis séricos de gonadotrofinas e esteróides sexuais, e a expressão de genes chaves na esteroidogênese ovariana (Cyp17a1 e Lhr). Foram utilizadas 30 ratas alocadas em igual número em três grupos. De acordo com os grupos, os animais receberam por injeção subcutânea, entre 0-3 dias de vida, os seguintes compostos: propionato de testosterona (1,25 mg) (grupo Testosterona); benzoato de estradiol (0,5 mg) (grupo Estradiol); e veículo (0,1 mL) (grupo Controle). Os animais foram pesados semanalmente a partir do nascimento até os 90 dias de vida. Ao completarem 90 dias, os animais foram eutanasiados e tiveram coletados o hipotálamo, os ovários e o sangue. Foi avaliada a expressão dos genes Gnrh, Gnrhr, Kiss1, Kiss1r e Ar no hipotálamo e Cyp17a1 e Lhr no ovário por PCR quantitativo em Tempo Real. O ovário também foi analisado morfológica e morfometricamente após coloração com hematoxilina e eosina (H.E.) e o sangue foi utilizado para a dosagem sérica dos hormônios LH, FSH, estradiol, progesterona e testosterona. A análise estatística foi realizada utilizando a análise de variância (ANOVA) complementada pelo teste de comparações múltiplas de Tukey. Os animais dos grupos Estradiol e Testosterona apresentaram 11 anovulação e maior peso corporal em relação ao grupo Controle. O gene Kiss1 encontrou-se hipoexpresso nos grupos Estradiol e Testosterona em relação ao grupo Controle (p<0,0001). No grupo Estradiol, o gene Kiss1r apresentou expressão aumentada em relação ao grupo Controle (p<0,01). O gene Gnrh não exibiu variações de expressão entre os grupos. A expressão de Gnrhr exibiu aumento no grupo Testosterona em relação ao grupo Estradiol (p<0,004). No grupo Testosterona, o gene do receptor de androgênio (Ar) apresentou expressão aumentada em relação ao grupo Controle (p<0,04). Quanto ao nível sérico de LH, o grupo Testosterona exibiu aumento em relação aos grupos Estradiol e Controle (p<0,001). O grupo Estradiol exibiu maior concentração sérica de estradiol em relação ao grupo Testosterona (p<0,01). Os níveis séricos de FSH, progesterona e testosterona não exibiram diferenças significativas entre os grupos. A expressão do gene Lhr apresentou-se aumentada nos grupos Estradiol e Testosterona em comparação ao Controle (p<0,003). O gene Cyp17a1 apresentou-se hiperexpresso no grupo Estradiol em relação aos grupos Controle e Testosterona (p<0,0001 e p<0,001, respectivamente). A contagem diferencial de folículos pré-antrais e antrais não diferiu significativamente entre os grupos. Os ovários dos grupos Estradiol e Testosterona apresentaram ausência de corpos lúteos e exibiram aumento da área intersticial e diminuição do volume do núcleo das células intersticiais, sendo todas essas alterações estatisticamente significativas (p<0,0001). Conclui-se que o excesso de estradiol ou testosterona na vida neonatal altera a fisiologia do hipotálamo e do ovário e leva a anovulação de ratas na vida adulta / Polycystic ovary syndrome (PCOS) is the most common endocrine disorder in women at reproductive age. Its etiology is unknown, but recently, environmental factors, such as excess of sex steroids in early life, have been compared to PCOS origin. In female rats, the excess of androgens or estrogens in neonatal life alters the hypothalamus-pituitary-gonadal axis function and induces to reproductive and metabolic alterations like those observed in human PCOS. The aim of this study is to analyze the expression of genes compared to GnRH secretion control (Gnrh, Gnrhr, Kiss1, Kiss1r e Ar) in the hypothalamus of animal models of PCOS and to evaluate the correlation with ovarian morphology, serum levels of gonadotropins and sex steroids and expression of key genes in ovarian steroidogenesis (Cyp17a1 e Lhr). 30 rats were used and allocated equally into three groups. According to the groups, animals received a subcutaneous injection, between 0-3 days old, of the following compounds: testosterone propionate (1.25 mg) (Testosterone group); estradiol benzoate (0.5 mg) (Estradiol group) and vehicle (0.1 mL) (Control group). Animals were weighed weekly from birth until 90 days of life. At 90 days, the animals were euthanized and were collected the hypothalamus, the ovaries and blood. We evaluated the expression of genes Gnrh, Gnrhr, Kiss1, Kiss1r and Ar in hypothalamus and Lhr and Cyp17a1 in ovary, both by quantitative Real-Time PCR. The ovary was also analyzed morphologically and morphometrically after staining with hematoxylin and eosin (H.E.) and the blood was used for evaluation of serum levels of LH, FSH, estradiol, progesterone and testosterone. Statistical analysis was performed using analysis of variance (ANOVA) complemented by Tukeys multiple comparisons test. The animals of the Testosterone and Estradiol groups had anovulation and had higher body weight compared to the Control group. The Kiss1 gene was downregulated in Estradiol and 13 Testosterone groups in relation to the Control group (p<0.0001). In Estradiol group, Kiss1r gene expression was increased in relation to the control group (p<0.01). The Gnrh gene expression did not show variations between groups. The Gnrhr expression was increased in Testosterone group in relation to Estradiol group (p<0.004). In Testosterone group, the androgen receptor gene (Ar) showed an increased expression compared to the Control group (p<0.04). Testosterone group exhibited increased serum levels of LH in comparison to Control and Estradiol groups (p<0.001). Estradiol group exhibited higher serum levels of estradiol compared to Testosterone group (p<0.01). Serum levels of FSH, progesterone and testosterone exhibited no significant differences between groups. Lhr gene expression was increased in Estradiol and Testosterone groups in comparison to the control (p<0.003). The Cyp17a1 gene was upregulated in Estradiol group compared to Testosterone and Control groups (p<0.0001 and p<0.001, respectively). The differential counting of preantral and antral follicles did not differ significantly between groups. Ovaries of the animals of Estradiol and Testosterone groups showed absence of corpora lutea and exhibited increased interstitial area and reduced interstitial cells nuclear volume, and all of these changes were statistically significant (p<0.0001). It was concluded that excess of estradiol or testosterone in neonatal life alters the hypothalamic and ovarian physiology and leads to anovulation in rats at adulthood
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Increased behavioural and histological variability arising from changes in cerebrovascular anatomy of the Mongolian gerbil /Laidley, David T., January 2005 (has links)
Thesis (M.Sc.)--Memorial University of Newfoundland, 2005. / Bibliography: leaves 31-42.
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Salt sensitivity in C57 and eNOS knockout mice /Leonard, Allison M., January 2005 (has links)
Thesis (M.Sc.)--Memorial University of Newfoundland, 2005. / Bibliography: leaves 132-142.
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Effects of aerobic exercise on the asthmatic lungHewitt, Matthew M. January 2008 (has links) (PDF)
Thesis (Ph.D.)--University of Alabama at Birmingham, 2008. / Title from PDF title page (viewed on Feb. 4, 2010). Includes bibliographical references.
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Neurorestorative strategies involving neurogenesis, neuronal precursors and stem cells in animal models of Parkinson's diseaseZhao, Ming, January 2009 (has links)
Diss. (sammanfattning) Stockholm : Karolinska institutet, 2009. / Härtill 4 uppsatser.
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Histopathology of human age-related macular degeneration and the development of a novel animal modelMaloney, Shawn C. January 2007 (has links)
Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly worldwide. Due to the inadequacy of current pharmacotherapies, novel molecular targets must be sought as potential therapeutic candidates. Furthermore, there is a need for more efficient and cost-effective animal models of this pathology in order to accelerate in vivo investigations. / Our laboratory is in possession of human choroidal neovascular membranes which we examined for expression of cyclooxygenase (COX)-2. This expression was characterized in retinal pigment epithelial, vascular endothelial, and fibroblast cells and correlated with patient age. We also looked at the feasibility of creating a rabbit laser-injury model to adequately mimic human neovascular AMD. / Our results suggest that anti-COX-2 therapies may be beneficial to some patients with neovascular AMD. Moreover, there is strong potential for the development of clinically relevant choroidal neovascularization in rabbits using the laser-injury technique. This approach may yield a novel, cost-effective AMD model.
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MCMV induced cerebellar maldevelopmentKoontz, Thadeus B. January 2007 (has links) (PDF)
Thesis (Ph. D.)--University of Alabama at Birmingham, 2007. / Title from first page of PDF file (viewed Feb. 7, 2008). Includes bibliographical references.
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