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Microparticles (MPs) were long dismissed as "platelet-dust", cell debris, with no functional significance. Theyare anucleated vesicles (0.1μm to 1μm), enclosed in a membrane, secretedinto the circulation by cells during cell activation or apoptosis. They have now emerged as mediators and markers of inflammatory diseases and autoimmune disorders. They are distinctly different from exosomes and apoptotic bodiesand are released from nearly every cell type, the most abundant being platelets, leukocytes and endothelial cells. MPs can be detected using flow cytometry and more recently by nanotechnology, which is more accuratein detecting, quantifying and phenotypingMPs.MPs are instrumental in the pathogenesis of various cardiovascular diseases (thrombotic and atherosclerotic) through their pro-inflammatory and pro-coagulant properties. Their levels are significantly elevated in chronic inflammatory disorders such as rheumatoid arthritis and multiple sclerosis. However, increasing evidence suggests they also possess antiinflammatory and anti-coagulant roles which could confer immunoprotection. MPs transport various lipids, proteins, mRNA and microRNA (miRNA) which may influence activities of receiving cells. Particularly the miRNAspecies delivered by MPs have been shown to modulate inflammation.In vitro and in vivo studies are being conducted to bioengineer MPs to facilitate delivery of therapeutic compounds to desired location safely, specifically and more effectively, with fewer side effects. More research is required to understand the composition, origin, mechanisms of formation and release as well as their clearance from the circulation to pave way for gaining greater pharmacological benefits by controlling MP-mediated cellular responses.
Microparticles (MPs) were long dismissed as "platelet-dust", cell debris, with no functional significance. Theyare anucleated vesicles (0.1μm to 1μm), enclosed in a membrane, secretedinto the circulation by cells during cell activation or apoptosis. They have now emerged as mediators and markers of inflammatory diseases and autoimmune disorders. They are distinctly different from exosomes and apoptotic bodiesand are released from nearly every cell type, the most abundant being platelets, leukocytes and endothelial cells. MPs can be detected using flow cytometry and more recently by nanotechnology, which is more accuratein detecting, quantifying and phenotypingMPs.MPs are instrumental in the pathogenesis of various cardiovascular diseases (thrombotic and atherosclerotic) through their pro-inflammatory and pro-coagulant properties. Their levels are significantly elevated in chronic inflammatory disorders such as rheumatoid arthritis and multiple sclerosis. However, increasing evidence suggests they also possess antiinflammatory and anti-coagulant roles which could confer immunoprotection. MPs transport various lipids, proteins, mRNA and microRNA (miRNA) which may influence activities of receiving cells. Particularly the miRNAspecies delivered by MPs have been shown to modulate inflammation.In vitro and in vivo studies are being conducted to bioengineer MPs to facilitate delivery of therapeutic compounds to desired location safely, specifically and more effectively, with fewer side effects. More research is required to understand the composition, origin, mechanisms of formation and release as well as their clearance from the circulation to pave way for gaining greater pharmacological benefits by controlling MP-mediated cellular responses.
The aim of this study was to investigate cell source-dependent nucleic acids repertoire of diverse subpopulations of plasma extracellular vesicles (EVs). Blood plasma from nine healthy volunteers was used for the analysis. Samples of EVs were obtained by differential centrifugation of plasma. The application of high-sensitivity fluorescence-activated vesicles sorting (hs-FAVS) using fluorophore-conjugated anti-CD41-FITC (Fluorescein isothiocyanate) and anti-CD235a-PE antibodies allowed the isolation of three subpopulations of EVs, namely CD41+ CD235a-, CD41-CD235a+ and CD41-CD235a dim. The high purity (>97%) of the sorted subpopulations was verified by highsensitivity flow cytometry. Presence of nanosized objects in sorted samples was confirmed by combination of low-voltage scanning electron microscopy and dynamic light scattering. The amount of material in sorted samples was enough to perform Quantitative polymerase chain reaction (qPCR)-based nucleic acid quantification. The most prominent differences in the nucleic acid repertoire were noted between CD41+ CD235-vs. CD41-CD235a+ vesicles: the former contained significantly (p = 0.004) higher amount of mitochondrial DNA, and platelet enriched miR-21-5p (4-fold), miR-223-3p (38-fold) and miR-199a-3p (187-fold), but lower amount of erythrocyte enriched miR-451a (90-fold). CD41-CD235a+ and CD41-CD235a dim vesicles differed in levels of miR-451a (p = 0.016) and miR-21-5p (p = 0.031). Nuclear DNA was below the limit of detection in all EV subpopulations. The hs-FCM-based determination of the number of sorted EVs allowed the calculation of per single-event miRNA concentrations. It was demonstrated that the most abundant marker in CD41+ CD235a-subpopulation was miR-223-3p, reaching 38.2 molecules per event. In the CD41-CD235+ subpopulation, the most abundant marker was miR-451a, reaching 24.7 molecules per event. Taken together, our findings indicate that erythrocyte-and platelet-derived EVs carry different repertoires of nucleic acids, which were similar to the composition of their cellular sources.
Τα μικροσωμάτια είναι μεμβρανικής-προέλευσης κυστίδια, που απελευθερώνονται μετά από κυτταρική διέγερση ή απόπτωση. Επιδεικνύουν προπηκτική δραστηριότητα (ΠΔΜ) και εμπλέκονται στην παθογένεση των αγγειακού υπόβαθρου επιπλοκών της κύησης. Αυτή η μελέτη ελέγχου περιπτώσεων συγκρίνει την προπηκτική δραστηριότητα των μικροσωματίων μεταξύ των εγκύων γυναικών με IUGR κυήσεις και των φυσιολογικών κυήσεων. ΜέθοδοιΤριάντα πέντε IUGR κυήσεις [ασύμμετρου τύπου (IUGRa), n=15; συμμετρικού (IUGRs), n=20] και 35 φυσιολογικές κυήσεις (μάρτυρες) μεταξύ 26 και 37 εβδομάδων, από τριτοβάθμιο νοσοκομείο εμπεριέχονται στην μελέτη. Όλα τα έμβρυα είχαν φυσιολογική ανατομία στον υπέρηχο. Λήψη ιστορικού, κλινική, βιοχημική και υπερηχογραφική εξέταση περιλήφθηκαν σε όλες τις έγκυες γυναίκες. Η ΠΔΜ υπολογίστηκε με το Zymuphen MP-Activity Kit (Hyphen BioMed, France). Το εύρος αναφοράς ήταν < 5 νανομοριακής ισοδύναμης φωσφατιδυλσερίνης (eq nM PS) και το όριο ανίχνευσης ήταν 0.05 Eq nM PS. Μια καμπύλη λειτουργικού χαρακτηριστικού δέκτη (ROC) κατασκευάστηκε για να αναλύσει την ακρίβεια της ΠΔΜ και να διακρίνει τις IUGR από τις απλές κυήσεις. Χρησιμοποιήθηκε ένα σταδιακό μοντέλο πολλαπλής γραμμικής παλινδρόμησης για την πρόβλεψη της τιμής της ΠΔΜ με βάση μια σειρά προγνωστικών παραγόντων. Αποτελέσματα. Οι IUGR κυήσεις είχαν υψηλότερο ΠΔΜ σε σύγκριση με τους μάρτυρες (p <0,001).Και οι δύο ομάδες IUGR (IUGRa και IUGRs) έδειξαν υψηλότερο ΠΔΜ συγκριτικά με τους μάρτυρες (p <0,001). Δεν διαπιστώθηκε διαφορά μεταξύ IUGRa και IUGRs (p = 0,069).Το μοντέλο πολλαπλής γραμμικής παλινδρόμησης εξήγησε το 82,9% της μεταβλητότητας της ΠΔΜ.CRP: C-αντιδρώσα πρωτεΐνη (mg / ml), ARΕDV: απών ή αναστρέψιμος τελικός διαστολικός όγκος, SGOT: γλουταμινική οξαλοξική τρανσαμινάση ορού (IU / ml). Συμπεράσματα. Τα μικροσωμάτια πιθανώς εμπλέκονται στην IUGR παθολογία καθώς IUGR κυήσεις έχουν υψηλότερη προπηκτική δραστηριότητα μικροσωματιδίων σε σύγκριση με τις μη επιπλεκόμενες κυήσεις.
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