BackgroundMesenchymal stem cells have been shown therapeutic in various neurological disorders. Recent studies support the notion that the predominant mechanism by which MSCs act is through the release of extracellular vesicles (EVs). EVs seem to have similar therapeutic activity as their cellular counterparts and may represent an interesting alternative standalone therapy for various diseases. The aim of the study was to optimize the method of EV imaging to better understand therapeutic effects mediated by EVs.MethodsThe fluorescent lipophilic stain PKH26 and superparamagnetic iron oxide nanoparticles conjugated with rhodamine (Molday ION Rhodamine B™) were used for the labeling of vesicles in human bone marrow MSCs (hBM-MSCs). The entire cycle from intracellular vesicles to EVs followed by their uptake by hBM-MSCs has been studied. The identity of vesicles has been proven by antibodies against: anti-CD9, -CD63, and -CD81 (tetraspanins). NanoSight particle tracking analysis (NTA), high-resolution flow cytometric analysis, transmission electron microscopy (TEM), ELYRA PS.1 super-resolution microscopy, and magnetic resonance imaging (MRI) were used for the characterization of vesicles.ResultsThe PKH26 and Molday ION were exclusively localized in intracellular vesicles positively stained for EV markers: CD9, CD63, and CD81. The isolated EVs represent heterogeneous population of various sizes as confirmed by NTA. The TEM and MRI were capable to show successful labeling of EVs using ION. Co-culture of EVs with hBM-MSCs revealed their uptake by cells in vitro, as visualized by the co-localization of PKH26 or Molday ION with tetraspanins inside hBM-MSCs.ConclusionPKH26 and Molday ION seem to be biocompatible with EVs, and the labeling did not interfere with the capability of EVs to re-enter hBM-MSCs during co-culture in vitro. Magnetic properties of IONs provide an additional advantage for the imaging of EV using TEM and MRI.

The compositions of free phenolic acids in rapeseed flours of diverse origin and white mustard were highly variable but represented less than 10% of the total phenolic acids. Phenolic acids released from hydrolysis of soluble esters constituted the major fraction, with Polish varieties having higher levels than a Canadian variety or white mustard. Yellow Sarson contained low levels of phenolic acids. Sinapic acid isomers constituted over 94% of the 13 phenolic acids found in the rapeseed varieties. Only traces of several phenolic acids appeared to be structurally bound to rapeseed and mustard proteins and carbohydrates.

This paper surveys the literature concerning biological properties of rapeseed glucosinolates, chiefly the goitrogenic activity of these compounds and their influence on the morphological and histological abnormalities of internal organs in animals. An attempt has been made to establish threshold glucosinolate levels in diet which trigger the onset or increase of internal organs impairment in animals depending on their species and breeding.

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