Dataset Information

Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine

ABSTRACT: Extracellular vesicles (EVs) have a great potential in clinical applications, however their isolation from different body fluids and their characterisation are not yet optimal or standardised. Here we report the results of examining the performance of ultrafiltration combined with size exclusion chromatography (UF-SEC) to isolate EVs from urine. The results reveal that UF-SEC is a fast method and provides high purity. Furthermore, we introduce asymmetrical-flow field-flow fractionation coupled with a UV detector and multi-angle light-scattering detector (AF4/UV-MALS) as a characterisation method and compare it with the applicabilities and limitations of current methods. We demonstrate that AF4/UV-MALS is a straightforward characterisation method for urinary EVs.

OTHER RELATED OMICS DATASETS IN: MTBLS291MTBLS397MTBLS396MTBLS462MTBLS1783MTBLS322MTBLS1103MTBLS619MTBLS832

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Urine

SUBMITTER: Kurt Boonen

LAB HEAD: Inge Mertens

PROVIDER: PXD009709 | Pride | 2018-07-04

REPOSITORIES: Pride

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Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine.

Oeyen Eline E Van Mol Kurt K Baggerman Geert G Willems Hanny H Boonen Kurt K Rolfo Christian C Rolfo Christian C Pauwels Patrick P Jacobs An A Schildermans Karin K Cho William C WC Mertens Inge I

Journal of extracellular vesicles 20180703 1

Extracellular vesicles (EVs) have a great potential in clinical applications. However, their isolation from different bodily fluids and their characterisation are currently not optimal or standardised. Here, we report the results of examining the performance of ultrafiltration combined with size exclusion chromatography (UF-SEC) to isolate EVs from urine. The results reveal that UF-SEC is an efficient method and provides high purity. Furthermore, we introduce asymmetrical-flow field-flow fractio ...[more]

PMID: 29988836

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Project description:The activities of the dithiolethione analogs, D3T, OLT, and TBD are pharmacologically well-understood. These compounds act as chemopreventive agents whose ability is to block or diminish early stages of carcinogenesis. In addition, the three compounds are classified as monofunctional Phase II enzyme inducers and activate the same pathway, namely the Keap1-Nrf2 signal pathway. The three dithiolethiones were showed to ameliorate the AFB1-induced toxicity through increasing phase II enzymes including glutathione S-transferase (GST). The parent D3T was observed to be the most potent chemoprotective agent. Oltipraz, a clinically approved drug, has been shown to exhibit less efficacy than its analogs for inhibition of aflatoxin-induced hepatic foci.TBD was suggested to be better than OLT as a chemopreventive agent because of its reduced toxicity profile. Thus based on gene expressions, our goal is to study the closely related structure activities that lie within the three compounds and the activity of OLT versus TBD at the 5 position of their dithiole rings and to determine whether the activity of TBD is closely related to OLT or its parent D3T. Experiment Overall Design: Male Fischer F344 rats (80-100g) were obtained from Harlan at 6 wk of age. General procedures for animal care and housing were in accordance with the Guide for the Care and use of Laboratory Animals, National Research Council, 1996. The animals were singly housed in polycarbonate cages in a temperature (22 ± 1°C)- and humidity (30-70%)-controlled room with a 14:10-h light-dark cycle, respectively. Rat AIN 76A chow (Harlan Teklad) and tap water were provided at libitum. Sixteen animals were randomly assigned into four groups (n=16) and treated by gavage with 100?l of either vehicle (saturated sucrose), 3H-1,2-dithiole-3-thione (D3T), (0.3 mmol/kg body wt), 4-methyl-5-pyrazinyl-3H-1,2-dithiole-3-thione (oltipraz-OLT), (0.3 mmol/kg), and 5-tert-butyl-3H-1,2-dithiole-3-thione (TBD) , (0.3mmol/kg body wt), once every other day for 5 days. Animals were killed 24h after the third dose. Liver tissues were harvested and snap frozen. The protocol for this study was approved by the University of Memphis Animal Care Committee and Johns Hopkins University.

Dataset Information

Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine

Publications

Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine.

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