Project description:Ionizing Radition is known to cause cell damage. Human peripheral blood mononuclear cells have long been used to study radiation induced gene expression profiling. Whithin this study we evaluated gene and microRNA expression alterations of human PBMC irradiated with 60 Gy g-ray. Cells were cultured for 2, 4 and 20h after irradiation before RNA was isolated and Agilent Human microRNA Microarrays was performed. Radiation induced gene expression in human peripheral blood mononuclear cells were measured at 2, 4 and 20 hours after exposure to doses of 60 Gy g-rays. Non radiated cell of each donor and time point were used as contol cells. Four independent experiments were performed at each time (2, 4, or 20 hours and 0 h - only 1 sample per donor) using 4 different donors.
Project description:Ionizing Radition is known to cause cell damage. Human peripheral blood mononuclear cells have long been used to study radiation induced gene expression profiling. Within this study we evaluated gene and microRNA expression alterations of human PBMC irradiated with 60 Gy g-ray. Cells were cultured for 2, 4 and 20h after irradiation before RNA was isolated and Agilent whole human GenomeOligo Microarray was performed. Radiation induced gene expression in human peripheral blood mononuclear cells were measured at 2, 4 and 20 hours after exposure to doses of 60 Gy g-rays. Non radiated cell of each donor and time point were used as contol cells. Four independent experiments were performed at each time (2, 4, or 20 hours and 0 h - only 1 sample per donor) using 4 different donors.
Project description:Ionizing Radition is known to cause cell damage. Human peripheral blood mononuclear cells have long been used to study radiation induced gene expression profiling. Whithin this study we evaluated gene and microRNA expression alterations of human PBMC irradiated with 60 Gy g-ray. Cells were cultured for 2, 4 and 20h after irradiation before RNA was isolated and Agilent Human microRNA Microarrays was performed.
Project description:Ionizing Radition is known to cause cell damage. Human peripheral blood mononuclear cells have long been used to study radiation induced gene expression profiling. Within this study we evaluated gene and microRNA expression alterations of human PBMC irradiated with 60 Gy g-ray. Cells were cultured for 2, 4 and 20h after irradiation before RNA was isolated and Agilent whole human GenomeOligo Microarray was performed.
Project description:For around ten years, microarrays have been suggested for the diagnosis of ionizing radiation exposure. We assessed for the first time the relevance of gene expression profiling in a real accidental case. This study was performed on peripheral blood mononuclear cells of 41 potential victims. The different strategies of analysis highlighted a huge effect of the blood sample handling on the gene expression profiles. This effect was so high that it could mask specific modulations as a potential effect of ionizing radiation exposure. Thus, we assessed a new way of blood sampling adapted to gene expression analysis: PAXgene. With this method, more than 70% of the modulations of gene expression induced 3 hours after an ex vivo exposure to 0.5 Gy were preserved even in a 24-hour delayed analysis (as for transportation of blood sample from the accident location to the laboratory). We validated a new methodology in order to propose a new strategy of blood sampling and handling for gene profiling. This system could be used in case of accidental overexposure to study whether gene expression is a relevant biomarker of ionizing radiation exposure. Radiation induced gene expression in human blood was measured at 3 hours after exposure to doses of 0 and 0.5 grays. Following the incubation of 3 hours at 37°C, the RNA extractions were performed either immediately or 24h later (as for transportation of blood sample at room temperature). Two different blood preservation methods were compared: classical anticoagulant and PAXgene Blood RNA System. Venous blood samples of 6 donors were used (3 for anticoagulant study, 3 for Paxgene study). Each sample was hybridized twice.
Project description:Genome-Scale draft model for Human Peripheral Blood Mononuclear Cells (PBMCs). A GEM for PBMCs was developed by applying the INIT
algorithm on Human Metabolic Reconstruction (HMR 2.0) as a template model. GEMs were contextualised/ constrained for different conditions using expression datasets. The gene/transcript expression data obtained from PBMCs of Type 1 Diabetes progressors, non-progressors, and healthy controls were employed to score each reaction of HMR 2.0. For further detail please refer to Electronic Supplementary Information of Sen et.al, Metabolic alterations in immune cells associate with progression to type 1 diabetes, Diabetologia, 15/01/2020, (https://doi.org/10.1007/s00125-020-05107-6).
Project description:For around ten years, microarrays have been suggested for the diagnosis of ionizing radiation exposure. We assessed for the first time the relevance of gene expression profiling in a real accidental case. This study was performed on peripheral blood mononuclear cells of 41 potential victims. The different strategies of analysis highlighted a huge effect of the blood sample handling on the gene expression profiles. This effect was so high that it could mask specific modulations as a potential effect of ionizing radiation exposure. Thus, we assessed a new way of blood sampling adapted to gene expression analysis: PAXgene. With this method, more than 70% of the modulations of gene expression induced 3 hours after an ex vivo exposure to 0.5 Gy were preserved even in a 24-hour delayed analysis (as for transportation of blood sample from the accident location to the laboratory). We validated a new methodology in order to propose a new strategy of blood sampling and handling for gene profiling. This system could be used in case of accidental overexposure to study whether gene expression is a relevant biomarker of ionizing radiation exposure.