Project description:The potential for extracellular RNA as a potential biomarker for human health and disease is well-established. This study explores how extracellular RNAs vary among healthy individuals and across different sample treatments.
Project description:Circulating extracellular RNAs (exRNAs) have the potential to serve as biomarkers for a wide range of medical conditions. However, limitations in existing exRNA isolation methods and a lack of knowledge on parameters affecting exRNA variability in human samples may hinder their successful discovery and clinical implementation. Using novel combinations of denaturants, reducing agents, proteolysis and revised organic extraction, we developed an automated, high-throughput approach for recovery of exRNAs and extracellular DNA (exDNA) from the same biofluid sample. We applied the method to characterize exRNAs from 312 plasma and serum samples collected from thirteen healthy volunteers at twelve time points over a two-month period.
Project description:An integrative analysis of human biofluid data in the exRNA Atlas revealed the existence of distinct extracellular RNA cargo types. To determine whether different RNA isolation kits biased detection of certain exRNA cargo types, an integrative analysis was performed using pooled plasma and serum samples, where 10 different RNA isolation kits were applied.
Project description:An integrative analysis of human biofluid data in the exRNA Atlas revealed the existence of distinct extracellular RNA cargo types. In order to detect differences in density between cargo types, cushioned density gradient ultracentrifugation (C-DGUC) of serum and plasma was performed using OptiPrem (TM) density gradient.
Project description:Blood as connective tissue potentially contain evidence of all processes occurring within the organism, at least in trace amounts. Because of their small size, peptides overcome cell membranes and epithelial barriers more freely than proteins. Among the peptides found in blood, there are both fragments of proteins secreted by various tissues and performing their function in plasma, as well as receptor ligands: hormones, cytokines, mediators of the cellular response. In addition, in minor amounts, there are disease peptide markers (for example, oncomarkers) and even foreign peptides related to pathogenic organisms and infection agents. To characterize the normal peptidome LC-MS/MS analysis was carried out of blood serum and plasma samples taken from 20 healthy donors on TripleTOF 5600+ mass-spectrometer. Sample preparation was carried out based on our previously developed method of peptide desorption from the surface of abundant blood plasma proteins followed by standard chromatographic steps.
Project description:An integrative analysis of human biofluid data in the exRNA Atlas revealed the existence of distinct extracellular RNA cargo types. To gain further insight on the biological nature of these cargo types, we correlated exRNA Atlas cargo profiles with a variety of other RNA-seq profiles. This study focuses on lipoprotein particle (LPP) exRNA profiles obtained via sequential density ultracentrifugation (SD-UC) and fast protein liquid chromatography (FPLC).