Project description:Human aortic endothelial cells were grown in culture until confluent. In three experiments using cells derived from three separate donors confluent cultures were incubated for 6 h with contol medium, or medium containing either extracts of oligomeric procyanidins from cranberry juice or red wine, or a procyanidin-rich grape seed extract. At the end of the 6 h treatment period conditioned media samples were retained for immunoassay of secreted peptides and proteins, and RNA was extracted for microarray analysis. Keywords: Comparative analysis of procyandins on vascular endothelial gene expression
Project description:Human aortic endothelial cells were grown in culture until confluent. In three experiments using cells derived from three separate donors confluent cultures were incubated for 6 h with contol medium, or medium containing either extracts of oligomeric procyanidins from cranberry juice or red wine, or a procyanidin-rich grape seed extract. At the end of the 6 h treatment period conditioned media samples were retained for immunoassay of secreted peptides and proteins, and RNA was extracted for microarray analysis. Experiment Overall Design: Each experiment used cells from one donor. Treatment conditions were: control medium, cranberry extract (CRE), grape seed extract (GSE), and red wine extract (RWE).
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:Analysis of ex vivo isolated lymphatic endothelial cells from the dermis of patients to define type 2 diabetes-induced changes. Results preveal aberrant dermal lymphangiogenesis and provide insight into its role in the pathogenesis of persistent skin inflammation in type 2 diabetes. The ex vivo dLEC transcriptome reveals a dramatic influence of the T2D environment on multiple molecular and cellular processes, mirroring the phenotypic changes seen in T2D affected skin. The positively and negatively correlated dLEC transcripts directly cohere to prolonged inflammatory periods and reduced infectious resistance of patients´ skin. Further, lymphatic vessels might be involved in tissue remodeling processes during T2D induced skin alterations associated with impaired wound healing and altered dermal architecture. Hence, dermal lymphatic vessels might be directly associated with T2D disease promotion. Global gene expression profile of normal dermal lymphatic endothelial cells (ndLECs) compared to dermal lymphatic endothelial cells derived from type 2 diabetic patients (dLECs).Quadruplicate biological samples were analyzed from human lymphatic endothelial cells (4 x diabetic; 4 x non-diabetic). subsets: 1 disease state set (dLECs), 1 control set (ndLECs)
Project description:Analysis of gene expression in primary cultured human aortic endothelial cells (HAECs) and primary cultured human aortic smooth muscle cells (SMCs) with or without H2O2 or Collagen tripeptide (CTP)
Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
Project description:Kynureninase is a member of a large family of catalytically diverse but structurally homologous pyridoxal 5'-phosphate (PLP) dependent enzymes known as the aspartate aminotransferase superfamily or alpha-family. The Homo sapiens and other eukaryotic constitutive kynureninases preferentially catalyze the hydrolytic cleavage of 3-hydroxy-l-kynurenine to produce 3-hydroxyanthranilate and l-alanine, while l-kynurenine is the substrate of many prokaryotic inducible kynureninases. The human enzyme was cloned with an N-terminal hexahistidine tag, expressed, and purified from a bacterial expression system using Ni metal ion affinity chromatography. Kinetic characterization of the recombinant enzyme reveals classic Michaelis-Menten behavior, with a Km of 28.3 +/- 1.9 microM and a specific activity of 1.75 micromol min-1 mg-1 for 3-hydroxy-dl-kynurenine. Crystals of recombinant kynureninase that diffracted to 2.0 A were obtained, and the atomic structure of the PLP-bound holoenzyme was determined by molecular replacement using the Pseudomonas fluorescens kynureninase structure (PDB entry 1qz9) as the phasing model. A structural superposition with the P. fluorescens kynureninase revealed that these two structures resemble the "open" and "closed" conformations of aspartate aminotransferase. The comparison illustrates the dynamic nature of these proteins' small domains and reveals a role for Arg-434 similar to its role in other AAT alpha-family members. Docking of 3-hydroxy-l-kynurenine into the human kynureninase active site suggests that Asn-333 and His-102 are involved in substrate binding and molecular discrimination between inducible and constitutive kynureninase substrates.
Project description:We isolated and cultured human aortic valvular endothelial cells from patients, and performed RNA sequencing of human aortic valvular endothelial cells transfected with siRNA targeting PPARG or negative control siRNA and treated with or without oxLDL.