Vascular endothelial growth factor-A (VEGF) is required for endothelial cell differentiation and survival. To investigate the renoprotective properties of VEGF in diabetes an inducible Cre-loxP gene targeting system was used to excise VEGF from podocytes of adult mice (VEGFKO). Diabetes was induced by streptozotocin (STZ) at 2.5 weeks of age and VEGFKO was induced by doxycycline (dox) at 3-4 weeks of age. Blood and urine were collected weekly to monitor for hyperglycaemia and proteinuria, respectively. Mice were dissected 8 weeks after diabetes induction or earlier if morbidly ill; twenty percent of the mice in the DM+VEGFKO group died before the surrogate endpoint. Glomerular VEGF mRNA expression was decreased in VEGFKO mice compared to controls. However, DM+VEGFKO mice had significantly greater proteinuria, degrees of glomerular sclerosis, and glomerular cell apoptosis. These results confirm that VEGF is normally upregulated in diabetes but reducing VEGF expression in diabetes causes severe kidney injury.

Orientador: Carlos Magno Castelo Branco Fortaleza / Resumo: Atualmente, estamos diante de uma notável presença de isolados de enterobactérias resistentes aos carbapenêmicos em um hospital público do município de Bauru-SP desde outubro de 2012. No entanto, não estão disponíveis estudos relacionando a epidemiologia e os fatores associados à aquisição de tais isolados. Este estudo teve como objetivo identificar fatores de risco para aquisição de Enterobactérias Resistentes aos Carbapenêmicos (CRE) em pacientes internados no Hospital Estadual Bauru, os fatores associados ao desenvolvimento de quadro infeccioso em uma coorte de pacientes colonizados por CRE e os fatores preditores de óbito. Foram incluídos pacientes do local de estudo que apresentaram colonização do trato digestório por CRE, de outubro de 2012 a dezembro de 2016, dos quais foram levantados dados clínicos e demográficos. Os isolados foram identificados por métodos fenotípicos e foram testadas as suscetibilidades por concentração inibitória mínima (MIC). Realizamos um estudo de caso-controle que incluiu 427 casos e igual número de controles. Os fatores de risco observados foram queimadura (HR 3,91; IC95% 2,36-6,46; p=<0,001), índice de Charlson (HR 1,12; IC95% 1,05-1,20; p=<0,001), uso prévio de esteróides (HR 2,79; IC95% 1,94-4,02; p=<0,001) e antimicrobianos como as penicilinas/inibidores de beta-lactamases (HR 2,01; IC95% 1,43-2,82; p=<0,001), cefalosporinas de 3ª. e 4ª. gerações (HR 2,45; IC95% 1,75-3,44; p=<0,001), quinolonas (HR 1,70; IC95% 1,75-2,45; p=0,003) e anaero... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Currently, we are facing a remarkable presence of isolates of carbapenem-resistant enterobacteriacea in Bauru city public hospital, São Paulo state, Brazil, since october 2012. However no studies are available relating the epidemiology and the factors associated with acquisition of such isolates. The purpose this study was to identify risk factors for acquisition of Carbapenem Resistant Enterobacteriaceae in patients hospitalized at the Bauru State Hospital, factors associated with the development of infectious disease in a cohort of patients colonized by CLP and factors predicting death. We included patients from the study site who presented colonization of the digestive tract by CRE, from October 2012 to December 2016, from which clinical and demographic data were collected. Isolates were identified by phenotypic methods and susceptibilities were tested by minimum inhibitory concentration (MIC). We performed a case-control study that included 427 cases and an equal number of controls. The risk factors observed were burn (HR 3.91, 95% CI 2.36-6.46, p = 0.001), Charlson index (HR 1.12, 95% CI 1.05-1.20, p = <0.001), previous use of steroids (HR 2.79, 95% CI 1.94-4.02, p = <0.001) and antimicrobials such as penicillins / beta-lactamase inhibitors (HR 2.01, 95% CI, 43-2.82, p = <0.001), cephalosporins of 3rd. and 4ª. (HR 2.45, IC 95% 1.75-3.44, p = 0.001), quinolones (HR 1.70, IC 95% 1.75-2.45, p = 0.003) and anaerobicides (HR 1, 63, 95% CI 1.04-2.56, p = 0.03). The cohort stud... (Complete abstract click electronic access below) / Doutor

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L'élastine est la protéine de la matrice extracellulaire responsable de l'élasticité des tissus. Elle est synthétisée en abondance dans les tissus soumis à de fortes contraintes mécaniques tels que la peau, les poumons, et les artères. Ce biopolymère présente une demi-vie longue de 70 ans et est par conséquent sujet aux altérations liées à l'âge. De plus, l'élastine est dégradée lors de la progression tumorale, notamment celle du mélanome. Sa dégradation entraîne la libération de peptides dérivés de l'élastine (EDP) dotés d'activités biologiques propres et pouvant être impliqués dans l'invasion tumorale (prolifération, angiogenèse, sécrétion de protéases, invasion, survie). Nous avons été les premiers à démontrer que les EDP sont capables de fortement promouvoir le développement de mélanome in vivo dans un modèle murin au travers de l'augmentation de la prolifération et de l'invasion des cellules tumorales impliquant la collagènase-1 ou MMP-1. Ces données suggèrent ainsi que le récepteur de ces peptides, le complexe récepteur de l'élastine (CRE), pourrait constituer une cible thérapeutique innovante afin de lutter contre ce cancer pour lequel un manque flagrant de thérapies efficientes prévaut à l'heure actuelle. Dans le cadre de cette étude, nous avons développé différentes stratégies thérapeutiques basées sur le ciblage du récepteur CRE. Nous démontrons pour la première fois que le ciblage thérapeutique du CRE permet de limiter de manière efficace la progression du mélanome in vitro et in vivo dans un modèle murin. De plus, le vieillissement étant un des facteurs de risque majeur du mélanome, nous avons évalué l'impact de l'âge du stroma sur les effets des EDP et le développement tumoral associé. / Elastin is the extracellular matrix protein responsible for tissue elasticity. It is abundant in tissues experiencing important mechanical constraints such as skin, lungs and arteries. This biopolymer possesses a half-life of 70 years and, consequently, it is subjected to age-related alterations. Moreover, elastin is degraded during tumor progression, notably that of melanoma. Its degradation results in the release of elastin-derived peptides (EDP) which possess their own biological activities that can be linked to tumor invasion (proliferation, angiogenesis, proteases secretion, invasion, survival). We have been the first to show that EDP can promote melanoma development in an in vivo mouse model by raising the proliferation and invasion of tumor cells through collagenase-1 (MMP-1) up-regulation. These data thus suggest that the receptor for these peptides, the elastin receptor complex, could constitute a novel therapeutic target to fight against this type of cancer for which no therapies are efficient now. In this study, we have developed therapeutic strategies targeting the ERC complex. We show for the first time that the therapeutic targeting of the ERC could efficiently limit melanoma progression in an in vitro and in vivo murine model. Additionally, as aging is one of melanoma major risk factors, we have evaluated the impact of stromal age on EDP effects and the associated tumor development.

京都大学 / 0048 / 新制・課程博士 / 博士(生命科学) / 甲第22133号 / 生博第420号 / 新制||生||55(附属図書館) / 京都大学大学院生命科学研究科高次生命科学専攻 / (主査)教授 渡邊 直樹, 教授 千坂 修, 教授 原田 浩 / 学位規則第4条第1項該当 / Doctor of Philosophy in Life Sciences / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22366号 / 医博第4607号 / 新制||医||1043(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 林 康紀, 教授 岩田 想, 教授 高橋 淳 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

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New genome-editing approaches, such as the CRISPR/Cas system, have opened up great opportunities to insert or delete genes at targeted loci and have revolutionized genetics in model organisms like the zebrafish. The Cre-loxp recombination system is widely used to activate or inactivate genes with high spatial and temporal specificity. Using a CRISPR/Cas9-mediated knock-in strategy, we inserted a zebrafish codon-optimized CreERT2 transgene at the otx2 gene locus to generate a conditional Cre-driver line.We chose otx2 as it is a patterning gene of the anterior neural plate that is expressed during early development. By knocking in CreERT2 upstream of the endogenous ATG of otx2, we utilized this gene’s native promoter and enhancer elements to perfectly match CreERT2 and endogenous otx2 expression patterns. Next, by combining this novel driver line with a Cre-dependent reporter line, we show that only in the presence of tamoxifen can efficient Cre-loxp-mediated recombination be achieved in the anterior neural plate-derived tissues like the telencephalon, the eye and the optic tectum. Our results imply that the otx2:CreERT2 transgenic fish will be a valuable tool for lineage tracing and conditional mutant studies in larval and adult zebrafish.

京都大学 / 新制・課程博士 / 博士(農学) / 甲第24669号 / 農博第2552号 / 新制||農||1099(附属図書館) / 学位論文||R5||N5450(農学部図書室) / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 菅瀬 謙治, 教授 小川 順, 教授 森 直樹 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM