Edible plant and fruit-derived nanovesicles (NVs) are membrane-enclosed particles with round-shape morphology and signaling functions, which resemble mammalian cell-derived extracellular vesicles. These NVs can transmit cross-kingdom signals as they contain bioactive molecules and exert biological effects on mammalian cells. Their properties and stability in the gastrointestinal tract suggest NVs as a promising nutraceutical tool. In this study, we have demonstrated for the first time the presence of NVs in olive vegetation water (OVW), a waste by-product generated during olive oil production. Biophysical characterization by scanning electron microscopy, cryo-transmission electron microscopy, and nanoparticle tracking analysis revealed the presence in OVW of NVs having size and morphology similar to that of vesicles isolated from edible plants. Integrated lipidomic, metabolomic, and proteomic analyses showed that OVW-NVs carry a set of lipids, metabolites and proteins which have recognized antioxidant and anti-inflammatory activities. The nature of biomolecules identified in OVW-NVs suggests that these vesicles could exert beneficial effects on mammalian cells and could be used in the nutraceutical and food industries. The successful isolation of OVW-NVs and the characterization of their features strengthen the idea that agricultural waste might represent a source of NVs having features similar to NVs isolated from edible plants/fruits.

Cereals are rich in nutrients and bioactive compounds; however, many of these, such as polyphenols, are bound to the cell wall matrix, limiting their bioavailability. This study investigated the use of fermentation to enhance the bioavailability of functional compounds in whole-wheat flour. Given the impact of microbial species on fermentation outcomes, various combinations of lactic acid bacteria and yeast strains were examined. The polyphenol and flavonoid content of different fermented flours was analyzed. Additionally, the antioxidant capacity was assessed using in vitro assays (DPPH, ORAC, and FRAP) and an ex vivo test with human erythrocytes. Fermentation significantly enhanced the release of bioavailable phenolic compounds and flavonoids, with the most significant increases reaching up to 3.4-fold and 2.64-fold, respectively. In particular, the findings highlight the capacity of flour fermented with a combination of K. humilis, F. sanfranciscensis, E. faecium, P. pentosaceus, and L. mesenteroides to enhance antioxidant activity in vitro and to protect human red blood cells from oxidative stress. Furthermore, fermentation increased the production of short-chain fatty acids, notably lactate and acetate, which are widely recognized for their gut health benefits. Overall, this study highlights the effectiveness of targeted fermentation in improving the bioactivity and antioxidant properties of whole-wheat flour.

Background/Objectives: Donkey milk (DM) has been considered a valuable alternative to human and bovine counterparts as well as to infant formulas. Milk extracellular vesicles (EVs) have been proposed to influence key biological processes. The purpose of this study is to provide a comprehensive characterization of the protein composition of extracellular vesicles (EVs) by extending quantitative proteomic comparisons to EVs derived from donkey colostrum (DC) and mature donkey milk (MDM). Methods: The EVs were isolated from DC and MDM samples, characterized, and subjected to proteomic analysis using the tandem mass tag-based quantitative approach. Results: In addition to typical milk proteins and EV markers, EVs from DC and MDM both contain components associated with the immune system, immune response, or promoting tissue repair, and assisting with communication between the infant and their environment. The EVs from DC were enriched in proteins associated with protein turnover, specific defense functions, and regenerative processes. Conclusions: Overall, the results can contribute to the broader characterization of the overall protein composition of DC and MDM and might help to predict the beneficial effects of the corresponding EVs on various mammalian cells. They may also provide valuable insights for the development of novel DM-based products for food, pharmaceutical, and biotechnological applications.