Made available in DSpace on 2014-12-02T11:16:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-08-08Bitstream added on 2014-12-02T11:21:18Z : No. of bitstreams: 1 000791765.pdf: 1879553 bytes, checksum: 60bed6e7ae5968f591931ddd672ad404 (MD5) / Os objetivos do trabalho foram avaliar o efeito de níveis e fontes de selênio na resposta imune de cães adultos, para isto cinco dietas foram avaliadas. Os tratamentos foram: CO - dieta basal com adição de selenito de sódio, totalizando 0,13 ppm de Se; SO33 - dieta basal + Se-levedura totalizando 0,33 ppm de Se; SO66 - dieta basal + Se-levedura totalizando 0,66 ppm de Se; SO99 - dieta basal + Se-levedura totalizando 0,99 ppm de Se; SI66 - dieta basal + selenito de sódio totalizando 0,66 ppm de Se. Os animais foram submetidos a um período de adaptação de 90 dias com a ração CO seguidos de 120 dias de fase experimental e recebendo as dietas testes, totalizando 210 dias. Amostras de sangue foram colhidas nos dias 0, 30, 60, 90 e 120 da fase experimental para: imunofenotipagem de leucócitos (IF), atividade fagocítica (AF) de monócitos e neutrófilos, índice de proliferação de células mononucleares do sangue periférico (IPCM), produção de intermediários reativos do oxigênio (H2O2) e do nitrogênio (NO-3). Para a determinação de anticorpos IgG e IgM específicos contra eritrócito de carneiro (SRBC) foram coletadas amostras de sangue 5, 10, 15 e 20 dias após inoculações de soluções de SRBC que ocorreram nos dias 60 e 90 da fase experimental. Fezes frescas foram colhidas para determinação da IgA fecal nos dias 0 e 120, e foi realizado o teste de hipersensibilidade cutânea tardia (DTH) aos 120 dias. O IPCM diminuiu em todos os tratamentos ao longo do experimento, porém aos 120 dias os animais do tratamento SO33 apresentaram a maior IPCM comparado aos demais (P<0,05). A AF de neutrófilos e monócitos foi menor aos 120 dias (P<0,0001) e houve tendência de maior AF por neutrófilos nos cães suplementados com 0,33 ppm de Se orgânico (P=0,0863). O número de linfócitos CD5+ aumentou após 60 dias de experimento para todos os tratamentos. Houve uma tendência para maior ... / The aims of this study were to evaluate the effects of selenium sources and levels on the immune response of adult dogs. Treatments were: CO - control diet with sodium selenite addition, totalizing 0.13 ppm of Se; SO33 - control diet + Se-yeast, totalizing 0.33 ppm of Se; SO66 - control diet + Se-yeast, totalizing 0.66 ppm of Se; and SO99 - control diet + Se-yeast, totalizing 0.99 ppm of Se; SI66 - control diet + sodium selenite, totalizing 0.66 ppm of Se. Animals underwent though an adaptation period of 90 days and received the CO diet; the experimental phase lasted 120 days when animals received the tested diets and samples were taken. Blood samples were collected on days 0, 30, 60, 90 e 120 of experimental phase to identify the leukocyte population (IF), to determine the phagocyte activity (FA) of neutrophils and monocytes, the proliferation of peripheral blood mononuclear cells (PIMC), the production of reactive oxygen (ROS) and nitrogen (NOS) intermediate species. Serum samples were also taken at 5, 10, 15 and 20 days after SRBC solution inoculation to determine the levels of IgG and IgM anti-SBRC. Fresh stool were obtained to evaluate the fecal IgA on days 0 and 120. Delayed-type hypersensitivity skin test (DTH) was conducted with the animals on day 120. The PICM reduced within time for all treatments, but on day 120th SO33-fed animals had the higher proliferation than the other treatments (P<0.05). The FA of neutrophils and monocytes was lower at 120th (P<0.0001) and a trend was observed for higher AF of neutrophils from SO33-fed dogs (P=0.0863). The population of CD5+ lymphocytes increased after 60 days and there was a trend for higher CD5+CD8+ lymphocytes count for animals fed CO, SO99 and SI66. The CD4+/CD5+ ratio was higher for animals fed SO33 and SO66 (P<0.0001). The values of IgG, IgM, IgA, ROS and NOS, and the DTH response were not influenced by treatments. Dose of ...

Orientador: José Roberto Sartori / Coorientador: Aulus Cavalieri Carciofi / Coorientador: Silvio Luis de Oliveira / Banca: Márcia de Oliveira Sampaio Gomes / Banca: Ricardo Souza Vasconcellos / Banca: Ricardo de Oliveira Orsi / Banca: Margarida Maria de Barros / Resumo: Os objetivos do trabalho foram avaliar o efeito de níveis e fontes de selênio na resposta imune de cães adultos, para isto cinco dietas foram avaliadas. Os tratamentos foram: CO - dieta basal com adição de selenito de sódio, totalizando 0,13 ppm de Se; SO33 - dieta basal + Se-levedura totalizando 0,33 ppm de Se; SO66 - dieta basal + Se-levedura totalizando 0,66 ppm de Se; SO99 - dieta basal + Se-levedura totalizando 0,99 ppm de Se; SI66 - dieta basal + selenito de sódio totalizando 0,66 ppm de Se. Os animais foram submetidos a um período de adaptação de 90 dias com a ração CO seguidos de 120 dias de fase experimental e recebendo as dietas testes, totalizando 210 dias. Amostras de sangue foram colhidas nos dias 0, 30, 60, 90 e 120 da fase experimental para: imunofenotipagem de leucócitos (IF), atividade fagocítica (AF) de monócitos e neutrófilos, índice de proliferação de células mononucleares do sangue periférico (IPCM), produção de intermediários reativos do oxigênio (H2O2) e do nitrogênio (NO-3). Para a determinação de anticorpos IgG e IgM específicos contra eritrócito de carneiro (SRBC) foram coletadas amostras de sangue 5, 10, 15 e 20 dias após inoculações de soluções de SRBC que ocorreram nos dias 60 e 90 da fase experimental. Fezes frescas foram colhidas para determinação da IgA fecal nos dias 0 e 120, e foi realizado o teste de hipersensibilidade cutânea tardia (DTH) aos 120 dias. O IPCM diminuiu em todos os tratamentos ao longo do experimento, porém aos 120 dias os animais do tratamento SO33 apresentaram a maior IPCM comparado aos demais (P<0,05). A AF de neutrófilos e monócitos foi menor aos 120 dias (P<0,0001) e houve tendência de maior AF por neutrófilos nos cães suplementados com 0,33 ppm de Se orgânico (P=0,0863). O número de linfócitos CD5+ aumentou após 60 dias de experimento para todos os tratamentos. Houve uma tendência para maior ... / Abstract: The aims of this study were to evaluate the effects of selenium sources and levels on the immune response of adult dogs. Treatments were: CO - control diet with sodium selenite addition, totalizing 0.13 ppm of Se; SO33 - control diet + Se-yeast, totalizing 0.33 ppm of Se; SO66 - control diet + Se-yeast, totalizing 0.66 ppm of Se; and SO99 - control diet + Se-yeast, totalizing 0.99 ppm of Se; SI66 - control diet + sodium selenite, totalizing 0.66 ppm of Se. Animals underwent though an adaptation period of 90 days and received the CO diet; the experimental phase lasted 120 days when animals received the tested diets and samples were taken. Blood samples were collected on days 0, 30, 60, 90 e 120 of experimental phase to identify the leukocyte population (IF), to determine the phagocyte activity (FA) of neutrophils and monocytes, the proliferation of peripheral blood mononuclear cells (PIMC), the production of reactive oxygen (ROS) and nitrogen (NOS) intermediate species. Serum samples were also taken at 5, 10, 15 and 20 days after SRBC solution inoculation to determine the levels of IgG and IgM anti-SBRC. Fresh stool were obtained to evaluate the fecal IgA on days 0 and 120. Delayed-type hypersensitivity skin test (DTH) was conducted with the animals on day 120. The PICM reduced within time for all treatments, but on day 120th SO33-fed animals had the higher proliferation than the other treatments (P<0.05). The FA of neutrophils and monocytes was lower at 120th (P<0.0001) and a trend was observed for higher AF of neutrophils from SO33-fed dogs (P=0.0863). The population of CD5+ lymphocytes increased after 60 days and there was a trend for higher CD5+CD8+ lymphocytes count for animals fed CO, SO99 and SI66. The CD4+/CD5+ ratio was higher for animals fed SO33 and SO66 (P<0.0001). The values of IgG, IgM, IgA, ROS and NOS, and the DTH response were not influenced by treatments. Dose of ... / Doutor

Orientador: Débora Cristina Damasceno / Resumo: Early embryos recovered from diabetic rats show increased number of blastomeres undergoing apoptosis and decreased total number of blastomeres, suggesting impaired litter size and newborn weight. This might increase the chances of metabolic disorders in adulthood of these animals. Since diabetes-induced hyperglycemia impairs the redox balance, the aim was to evaluate the embryonic oxidative stress status before the implantation in order to identify whether there are differences in levels of oxidative stress in early embryos from diabetic dams. Thus, we hypothesized the decreased cell number found in early embryos recovered from diabetic rats is due to the low embryonic ability to maintain their own redox balance. Our study shows that rats with streptozotocin-induced mild and severe diabetes present impaired redox status in early pregnancy. This maternal unbalance directly influences the embryonic redox status, reflecting the increased reactive oxygen species in the morula stage. We also showed that regardless of hyperglycemic level the embryos trigger defense mechanisms involved in the excessive free radical scavenging as an attempt to survive. Nevertheless, the success of this defense mechanism seems to depend on maternal hyperglycemia, highlighting the importance of the programmed pregnancy as well as appropriate medical care starting in early stages of human diabetic pregnancy. / Doutor

Orientador: José Corrêa de Lacerda Neto / Coorientador: Kamila Gravena / Banca: Annelise Carla Camplesi dos Santos / Banca: Alceu Afonso Jordão Junior / Resumo: A síndrome cólica um dos distúrbios de maior importância em equinos, a se destacar entre suas causas os quadros obstrutivos, decorrentes de corpos estranhos e ou enterólitos localizados no cólon menor. No desenvolvimento da cólica há diminuição do fluxo sanguíneo local ao qual se segue, quando de sua correção, a reperfusão sanguínea. A isquemia seguida de reperfusão é a causa do aumento da lesão degenerativa principal pós-cirúrgica. As lesões de reperfusão podem ser causadas pela produção e liberação de espécies reativas de oxigênio(EROs). Diante disto, objetivou-se no presente trabalho analisar moléculas decorrentes da oxidação celular e substâncias antioxidantes no soro e no tecido do cólon menor em equinos submetidos experimentalmente a obstrução intraluminal do cólon menor. Também foram avaliados parâmetros físicos frequência cardíaca (FC), frequência respiratória (FR), temperatura retal (Tr), motilidade, tempo de preenchimento capilar (TPC), coloração de mucosas, pulso digital e turgor cutâneo. Análises hematimétricas e leucométricas foram efetuadas em diferentes momentos. Análises no soro para determinação de malondialdeido (MDA), produto protéico de oxidação avançada (AOPP) (metabólitos da oxidação), glutationa (GSH) e vitamina E (Vit. E) (substâncias antioxidantes), foram realizadas em diferentes momentos: M0 (momento antes do procedimento anestésico e precurção da obstrução), M4 (4 horas, retirada da obstrução), M12, M24, M36, M48, M60 e M72 (correspondendo a 12, 24, 36, 48, 60 e 72 horas após a obstrução). No cólon menor foram obtidas amostras em M0 (antes da obstrução), após 4 horas de obstrução intestinal (M4) foram obtidas na região oral, obstruída e aboral. Após 72 horas de pós cirúrgico (M72) realizou-se nova coleta das regiões do cólon menor. Nestas amostras de tecido foram realizadas as analises de MDA, AOPP, GSH e catalase. Nos parâmetros... / Abstract: The syndrome colic one of the most important disorders in horses, stand out among its causes obstructive frames, resulting from foreign bodies and or enteroliths located in the lower colon. The development of colic there is reduced local blood flow which follows, when their correction, blood reperfusion. The ischemia followed by reperfusion is the cause of increased postoperative primary degenerative injury. Reperfusion injury can be caused by the production and release of reactive oxygen species (ROS). In view of this, the aim of the present study to analyze molecules resulting from cellular oxidation and antioxidants in serum and lower colonic tissue in horses submitted to experimental intraluminal obstruction of the lower colon. They also evaluated physical parameters heart rate (HR), respiratory rate (RR), rectal temperature (Tr), motility, capillary refill time (TPC), color of mucous membranes, digital pulse and skin turgor. Hematological analysis and leukocytes were made at different times. Analysis of serum for determination of malondialdehyde (MDA) and vitamin E (Vit E), protein product of advanced oxidation (AOPP) (oxidation metabolites), glutathione (GSH) (antioxidant flux), were held at different times: M0 (time before the anesthetic and procedure obstruction), M4 (4 hours, removal of the obstruction), M12, M24, M36, M48, M60 and M72 (corresponding to 12, 24, 36, 48, 60 and 72 hours after occlusion). In the lower colon samples were obtained from M0 (before blocking) after 4 hours of intestinal obstruction (M4) were obtained in the oral region and aboral obstructed. After 72 hours of post surgical (M72) held new collection regions of the lower colon. In these tissue samples were carried out analysis of MDA, AOPP, catalase and GSH. In physiological parameters we observed a significant increase in HR and Tr in M12, and an increase in RR in M24. Hematological evaluation showed a decrease compared to red blood cell count ... / Mestre

The cell possesses a number of vital mechanisms to respond to different stressors. Mitochondria are dynamic organelles which undergo constant changes in length, transport and inner membrane structure and curvature. Invaginations of this inner membrane, cristae, have been known to respond to the energetic state of mitochondria, but the regulation of these changes as well as the consequences thereof remain undetermined. We find that Optic Atrophy 1 (OPA1), a protein involved in inner membrane fusion and cristae maintenance during cell death, can respond to the energetic state of mitochondria and the cell. Moreover, OPA1-dependent changes in cristae structure are required for resistance to starvation induced cell death, proper functioning of the electron transport chain, for growth in galactose media and for maintenance of ATP synthase assembly. Interestingly, we demonstrate that select members of the mitochondrial solute carriers (SLC25A) interact with OPA1 and affect the response of OPA1 to substrate levels. Taken together, we propose an SLC25A-dependent role for OPA1 in sensing energy substrate availability and responding to alter cristae, bioenergetics and cellular survival. We also identified KIAA0664 as a novel OPA1-interacting protein, describe its subcellular localization and investigate its role in mitochondrial fusion and in mitochondrial localization. Finally, since both known carriers of mitochondrial glutathione were demonstrated to interact with OPA1, we investigated the role of OPA1 in cellular glutathione redox. OPA1 depleted cells demonstrated both increased total cellular glutathione and a shift in redox to its reduced form. The role of OPA1 in glutathione levels and redox ratios required GTPase activity, but surprisingly not fusion. Since glutathione is a master regulator of reactive oxygen species detoxification, these findings may shed light on the role of OPA1 in ROS-induced cell death pathways.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21998号 / 医博第4512号 / 新制||医||1038(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 柳田 素子, 教授 松田 文彦, 教授 木村 剛 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Protein degradation poses a significant challenge for the efficient production of recombinant proteins in plants, affecting the stability and yield of the recombinant protein. In this study the E. coli-derived enzyme glutathione reductase (GR) was transiently expressed in transgenic tobacco plants constitutively expressing the cysteine protease inhibitor OC-I and non-transgenic plants. A protein resembling the GR was detected in infiltrated leaves. Transiently expressing GR in transgenic N tabacum plants resulted in almost two fold significant increases in GR activity. Transgenic tobacco plants expressing the rice cysteine protease inhibitor OC-I had significantly lower cysteine protease activity when compared to non-transgenic tobacco plants. Lower cysteine protease activity in transgenic plants was directly related to higher GR activity and also higher GR amounts in transgenic plants. The study has demonstrated that OC-I is an effective companion protease inhibitor candidate with the potential to protect other high value proteins such as GR, from cysteine protease degradation. / Dissertation (MSc)--University of Pretoria, 2012. / Plant Science / unrestricted

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20974号 / 医博第4320号 / 新制||医||1026(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 増永 慎一郎, 教授 高田 穣, 教授 武田 俊一 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

No description available.

No description available.