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Residual matrix from different separation techniques impacts exosome biological activity.

ABSTRACT: Exosomes are gaining a prominent role in research due to their intriguing biology and several therapeutic opportunities. However, their accurate purification from body fluids and detailed physicochemical characterization remain open issues. We isolated exosomes from serum of patients with Multiple Myeloma by four of the most popular purification methods and assessed the presence of residual contaminants in the preparations through an ad hoc combination of biochemical and biophysical techniques - including Western Blot, colloidal nanoplasmonics, atomic force microscopy (AFM) and scanning helium ion microscopy (HIM). The preparations obtained by iodixanol and sucrose gradients were highly pure. To the contrary, those achieved with limited processing (serial centrifugation or one step precipitation kit) resulted contaminated by a residual matrix, embedding the exosomes. The contaminated preparations showed lower ability to induce NfkB nuclear translocation in endothelial cells with respect to the pure ones, probably because the matrix prevents the interaction and fusion of the exosomes with the cell membrane. These findings suggest that exosome preparation purity must be carefully assessed since it may interfere with exosome biological activity. Contaminants can be reliably probed only by an integrated characterization approach aimed at both the molecular and the colloidal length scales.

SUBMITTER: Paolini L

PROVIDER: S-EPMC4806376 | biostudies-other | 2016 Mar

REPOSITORIES: biostudies-other

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Project description:To identify a proper strategy for future feed-efficient pig farming, it is required to evaluate the ongoing selection scenarios. Tools are lacking for the evaluation of pig selection scenarios in terms of environmental impacts to provide selection guidelines for a more sustainable pig production. Selection on residual feed intake (RFI) has been proposed to improve feed efficiency and potentially reduce the associated environmental impacts. The aim of this study was thus to develop a model to account for individual animal performance in life cycle assessment (LCA) methods to quantify the responses to selection. Experimental data were collected from the fifth generation of pig lines divergently selected for RFI (low line, more efficient pigs, LRFI; high line, less efficient pigs, HRFI). The average feed conversion ratio (FCR) and daily feed intake of LRFI pigs were 7% lower than the average of HRFI pigs (P < 0.0001). A parametric model was developed for LCA based on the dietary net energy fluxes in a pig system. A nutritional pig growth tool, InraPorc®, was included as a module in the model to embed flexibility for changes in feed composition, animal performance traits and housing conditions and to simulate individual pig performance. The comparative individual-based LCA showed that LRFI had an average of 7% lower environmental impacts per kilogram live pig at farm gate compared to HRFI (P < 0.0001) on climate change, acidification potential, freshwater eutrophication potential, land occupation and water depletion. High correlations between FCR and all environmental impact categories (>0.95) confirmed the importance of improvement in feed efficiency to reduce environmental impacts. Significant line differences in all impact categories and moderate correlations with impacts (>0.51) revealed that RFI is an effective measure to select for improved environmental impacts, despite lower correlations compared to FCR. Altogether more optimal criteria for efficient environment-friendly selection can then be expected through restructuring the selection indexes from an environmental point of view.

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Residual matrix from different separation techniques impacts exosome biological activity.

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