Genomic profiling of human vascular cells identifies TWIST1 as a causal gene for common vascular diseases

Genome-wide association studies have identified multiple novel genomic loci associated with vascular diseases. Many of these loci are common non-coding variants that affect the expression of disease-relevant genes within coronary vascular cells. To identify such genes on a genome-wide level, we performed deep transcriptomic analysis of genotyped primary human coronary artery smooth muscle cells (HCASMCs) and coronary endothelial cells (HCAECs) from the same subjects, including splicing Quantitative Trait Loci (sQTL), allele-specific expression (ASE), and colocalization analyses. We identified sQTLs for TARS2, YAP1, CFDP1, and STAT6 in HCASMCs and HCAECs, and 233 ASE genes, a subset of which are also GTEx eGenes in arterial tissues. Colocalization of GWAS association signals for coronary artery disease (CAD), migraine, stroke and abdominal aortic aneurysm with GTEx eGenes in aorta, coronary artery and tibial artery discovered novel candidate risk genes for these diseases. At the CAD and stroke locus tagged by rs2107595 we demonstrate colocalization with expression of the proximal gene TWIST1. We show that disrupting the rs2107595 locus alters TWIST1 expression and that the risk allele has increased binding of the NOTCH signaling protein RBPJ. Finally, we provide data that TWIST1 expression influences vascular SMC phenotypes, including proliferation and calcification, as a potential mechanism supporting a role for TWIST1 in CAD

Large animal models of cardiovascular disease

The human cardiovascular system is a complex arrangement of specialized structures with distinct functions. The molecular landscape, including the genome, transcriptome and proteome, is pivotal to the biological complexity of both normal and abnormal mammalian processes. Despite our advancing knowledge and understanding of cardiovascular disease (CVD) through the principal use of rodent models, this continues to be an increasing issue in today's world. For instance, as the ageing population increases, so does the incidence of heart valve dysfunction. This may be because of changes in molecular composition and structure of the extracellular matrix, or from the pathological process of vascular calcification in which bone-formation related factors cause ectopic mineralization. However, significant differences between mice and men exist in terms of cardiovascular anatomy, physiology and pathology. In contrast, large animal models can show considerably greater similarity to humans. Furthermore, precise and efficient genome editing techniques enable the generation of tailored models for translational research. These novel systems provide a huge potential for large animal models to investigate the regulatory factors and molecular pathways that contribute to CVD in vivo. In turn, this will help bridge the gap between basic science and clinical applications by facilitating the refinement of therapies for cardiovascular disease. Copyright (c) 2016 John Wiley & Sons, Ltd

Endothelial phenotypes in a hypercholesterolemic swine model of early aortic valve sclerosis

The aortic valve of humans and swine is preferentially susceptible to AVS localized to the aortic side of the leaflets. The endothelium may play a role in establishing this pattern of disease by regulating valve permeability, inflammation, and lipid transport in aortic valves, as it does in arteries. However, there are few experimental studies of valve endothelial cell and molecular biology. Consequently, valve endothelial homeostasis in general and the role of endothelium in early valve pathogenesis is poorly understood, particularly in relation to the preferential sidedness of AVS. Most challenging is the determination of spatially-defined valvular cell phenotypes in situ/in vivo. To study the transition from normal valve to AVS, I proposed that a brief in vivo exposure to hypercholesterolemia (HC), a well-established risk factor for AVS, will elicit differential side-specific changes in endothelial phenotypes, define their spatial heterogeneity and reveal molecular mechanisms underlying the initiation of aortic valve disease. A global genomics approach was chosen to address this hypothesis. I further proposed progressive changes in endothelial phenotypes during an extended 6 month HC period, studies that are addressed at the level of differential in situ protein expression. HC was induced in adult male swine over 2 weeks. Sub-endothelial lipid in-sudation and occasional calcific nodules were noted exclusively on the aortic side of the leaflets. Side-specific endothelial transcript profiles were determined using custom-printed porcine oligomer microarrays and investigated using bioinformatics and pathway analyses. Aortic-side and ventricular-side endothelia were highly heterogeneous in their responses to systemic HC; specifically, the aortic side endothelium displayed heightened sensitivity. HC induced the differential expression of 1325 endothelial genes on the aortic side in contrast to 87 genes on the pathoprotected ventricular side. Detailed pathway analyses identified multiple genes of the annexinA2-, caspase3-, PPARγ-, and TNFα-related pathways as differentially expressed on the aortic side in response to HC (vs normocholesterolemia) and, by independent analyses, relative to the ventricular side during HC. The directions of pathway gene differential expression were consistent with a protective endothelial phenotype. Protein immunocytochemistry substantiated the genomics results at 2 weeks and showed that differential HC-induced endothelial phenotype expression persisted at 6 months

Endothelial phenotypes in a hypercholesterolemic swine model of early aortic valve sclerosis

The aortic valve of humans and swine is preferentially susceptible to AVS localized to the aortic side of the leaflets. The endothelium may play a role in establishing this pattern of disease by regulating valve permeability, inflammation, and lipid transport in aortic valves, as it does in arteries. However, there are few experimental studies of valve endothelial cell and molecular biology. Consequently, valve endothelial homeostasis in general and the role of endothelium in early valve pathogenesis is poorly understood, particularly in relation to the preferential sidedness of AVS. Most challenging is the determination of spatially-defined valvular cell phenotypes in situ/in vivo. To study the transition from normal valve to AVS, I proposed that a brief in vivo exposure to hypercholesterolemia (HC), a well-established risk factor for AVS, will elicit differential side-specific changes in endothelial phenotypes, define their spatial heterogeneity and reveal molecular mechanisms underlying the initiation of aortic valve disease. A global genomics approach was chosen to address this hypothesis. I further proposed progressive changes in endothelial phenotypes during an extended 6 month HC period, studies that are addressed at the level of differential in situ protein expression. HC was induced in adult male swine over 2 weeks. Sub-endothelial lipid in-sudation and occasional calcific nodules were noted exclusively on the aortic side of the leaflets. Side-specific endothelial transcript profiles were determined using custom-printed porcine oligomer microarrays and investigated using bioinformatics and pathway analyses. Aortic-side and ventricular-side endothelia were highly heterogeneous in their responses to systemic HC; specifically, the aortic side endothelium displayed heightened sensitivity. HC induced the differential expression of 1325 endothelial genes on the aortic side in contrast to 87 genes on the pathoprotected ventricular side. Detailed pathway analyses identified multiple genes of the annexinA2-, caspase3-, PPARγ-, and TNFα-related pathways as differentially expressed on the aortic side in response to HC (vs normocholesterolemia) and, by independent analyses, relative to the ventricular side during HC. The directions of pathway gene differential expression were consistent with a protective endothelial phenotype. Protein immunocytochemistry substantiated the genomics results at 2 weeks and showed that differential HC-induced endothelial phenotype expression persisted at 6 months

Sistema de reforzamiento exomuscular cervical para la práctica de rodeo chileno

Tesis (Diseñador Industrial)Objetivo General: Comprender las condiciones en las cuales se practica el rodeo Chileno en torno a la seguridad de un jinete que cabalga de manera no tradicional, y por lo tanto sufre de accidentes y lesiones atípicas en un entorno tradicionalista. Objetivos específicos: 1- Establecer cuáles son las condiciones en torno a la seguridad en las que compiten actualmente los jinetes. 2- Categorizar y numerar los grados de lesión que implican la práctica de este deporte. 3- Listar las causas asociadas a los accidentes que se producen durante una carrera. 4- Numerar los sistemas de seguridad que operan actualmente en el rodeo

Actividades acuáticas en niñas y niños de NB3 de establecimientos educacionales municipalizados del sector centro de la comuna de Hijuelas

Tesis (Educación Física)La Reforma educacional de 1990 ha introducido cambios innovadores en el modelo curricular donde en estos nuevos diseños, la definición de Objetivos Fundamentales y Contenidos Mínimos Obligatorios ha significado para todos los estudiantes de nuestro país poner en práctica un sistema en el que participan tanto el Estado como los mismos establecimientos educacionales. En el área de la Educación Física la reforma ha propuesto en sus Planes y Programas a partir de Quinto año básico (NB3) la inclusión de la natación dentro de sus contenidos, donde se formula como: práctica y desarrollo de las habilidades de flotar y nadar hasta lograr dominio: ejercitar principios y habilidades de seguridad sobrevivencia en el agua. Nos resulta interesante saber cómo se abordan las actividades acuáticas en los establecimientos educacionales a lo largo de Chile, especialmente en las zonas rurales alejadas del mar, donde no siempre existe una infraestructura adecuada para llevar a cabo esta innovadora y necesaria propuesta. Esta investigación está enfocada específicamente en verificar el desarrollo propuesto por los Planes y Programas en cuanto a la realización de la Natación como contenido en el nivel de NB3 de los establecimientos educacionales municipalizados que se ubican en el sector centro de la Comuna de Hijuelas, perteneciente a la Provincia de Quillota en la región de Valparaíso

Migraciones en las Américas: Reflexiones en relación a la agenda de seguridad

Considering the current state of migration in the Americas, a brief analysis is proposed in relation to the security agenda. We conclude that migratory flows are a matter of intermestic character because, on one hand are related with inherent aspects of the international system and on the other hand are linked with internal matters of sending and hosting countries.Considerando el estado actual de las migraciones en las Américas, se propone un breve análisis en relación a la agenda de seguridad. Se concluye que esta es una materia de carácter interméstico, pues vincula aspectos insoslayables al sistema internacional y asuntos de índole interna a nivel de países emisores y receptores de migrantes