Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

Paracrine roles of extracellular vesicles released by mouse mesoangioblasts

Extracellular vesicles (EV) represent an important mediator of cell-to-cell communication and are involved in both autocrine and paracrine signaling, with a critical role in a number of physiological and pathological conditions.1 The bioactive molecules contained within EV simultaneously activate several different pathways resulting in the synergistic stimulation of target cells. The discovery and characterization of EV have added a novel understanding to regenerative medicine, namely the finding that stem cells are an abundant source of EV.1-2 A6 mouse mesoangioblasts, vessel-associated multipotent progenitor stem cells that are capable of differentiating into different mesodermal cell types, are able to release in the extracellular environment membrane vesicles, which contain structural proteins, FGF-2 and the two gelatinases MMP2 and MMP9.3 Moreover, we have already demonstrated that EV released by these cells contain Hsp70 as a transmembrane protein, which is involved in an autocrine signaling responsible for increased cell migration.4 In this study we have investigated the possible paracrine effects of A6 derived EV with other cell types, and the effects of these interactions. Firstly, we have focused our attention on their interaplay capacity with human endothelial cells, which are induced to form capillary-like structures in vitro and to increase their motility. Furthermore, we have analyzed EV immunomodulatory effect on Jurkat lymphocytes, demonstrating that they are able to inhibit both their activation and proliferation. Finally, we have investigated the role of sugar residues on the membrane of A6 derived EV in their interaction with other cell types, by enzymatic removing of N-linked glycans on their membrane. In particular, PNGase F that is responsible for the cleavage between asparagine and GlcNAc in all types of glycan chains induces a substantial reduction in EV-target cell interaction. On the contrary, the use of EndoH, which is responsible for the cleavage between two residues of GlcNAc, increase target cell-EV interaction

H2O2 resistant mesoangioblast clone isolation with a distinct survival advantage in vitro and in vivo

The release of molecules from damaged tissues stimulates both resident and circulating stem cells to initiate a tissue repair programme. 1 However, during transplantation procedures the therapeutic efficacy of stem cells is compromised by reduced homing capability towards the target site.2,3 Furthermore, cell survival is very low and many studies focused on improving cell viability upon implantation. In this study, we performed in vitrosevere oxidative stress to select some more resistant mouse mesoangioblasts, vessel-associated progenitor stem cells endowed with the ability of multipotent mesoderm differentiation. We found that the selected subpopulation retains selfrenewal and myogenic differentiation capabilities under physiological growth conditions, showing, also, an enhancement in cell survival and migration capabilities under stress conditions respect to the unselected cells. In fact, following oxidative stress treatment the isolated cell subpopulation showed more resistance, survival and recovery properties. To evaluate whether or not the isolated cell clone showed selective advantages over the parental mesoangioblasts, we carried out in vivo experiments using immunocompromised dystrophic mice. We injected intramuscularly the Tibialis Anterior with both the selected cells and the parental cells. Actually resistant mesoangioblasts displayed markedly enhanced survival and integration capabilities into the host damaged skeletal muscle, displaying more than 70% increase in integration compared to the non-selected mesoangioblast cell population. In conclusion, the positive effects of sorting on mesoangioblast cells suggest that a selection step using oxidative stress preconditioning may provide a novel methodology to select for resistant cells that can be used in regenerative tissue applications to prevent high mortality rates upon transplantation

The effects of processing and mastication on almond lipid bioaccessibility using novel methods of in vitro digestion modelling and micro-structural analysis.

A number of studies have demonstrated that consuming almonds increases satiety but does not result in weight gain, despite their high energy and lipid content. To understand the mechanism of almond digestion, in the present study, we investigated the bioaccessibility of lipids from masticated almonds during in vitro simulated human digestion, and determined the associated changes in cell-wall composition and cellular microstructure. The influence of processing on lipid release was assessed by using natural raw almonds (NA) and roasted almonds (RA). Masticated samples from four healthy adults (two females, two males) were exposed to a dynamic gastric model of digestion followed by simulated duodenal digestion. Between 7·8 and 11·1 % of the total lipid was released as a result of mastication, with no significant differences between the NA and RA samples. Significant digestion occurred during the in vitro gastric phase (16·4 and 15·9 %) and the in vitro duodenal phase (32·2 and 32·7 %) for the NA and RA samples, respectively. Roasting produced a smaller average particle size distribution post-mastication; however, this was not significant in terms of lipid release. Light microscopy showed major changes that occurred in the distribution of lipid in all cells after the roasting process. Further changes were observed in the surface cells of almond fragments and in fractured cells after exposure to the duodenal environment. Almond cell walls prevented lipid release from intact cells, providing a mechanism for incomplete nutrient absorption in the gut. The composition of almond cell walls was not affected by processing or simulated digestion

Health effects of phloretin: from chemistry to medicine

Dihydrochalcones are a class of secondary metabolites, whose demand in biological and pharmacological applications is rapidly growing. Phloretin is one of the best known and abundant dihydrochalcone characterized by the presence of 2,6-dihydroxyacetophenone pharmacophore. It is a versatile molecule with anticancer, antiosteoclastogenic, antifungal, antiviral, anti-inflammatory, antibacterial and estrogenic activities and able to increase the fluidity of biological membranes and penetration of administered drugs. In this review we have performed a critical evaluation of available literature as far as phloretin beneficial effects and activation/block of intracellular signal cascade are of concern. In addition, we supply useful information on its chemical properties, sources and bioavailability

Solitary splenic metastasis from nasopharyngeal carcinoma: a case report and systematic review of the literature

Background: Solitary splenic metastases are a rare occurrence, and the nasopharyngeal carcinoma represents one of the most uncommon primary sources. The present study aimed to describe a rare case of a solitary single splenic metastasis from nasopharyngeal carcinoma and to assess the number of cases of isolated nasopharyngeal carcinoma metastases to the spleen reported in the literature. Main body: We describe the case of a 56-year-old man with a history of nasopharyngeal carcinoma and complete remission after chemo-radiotherapy. Three months after complete remission, positron emission tomography/ computed tomography scan revealed a hypermetabolic splenic lesion without increased metabolic activity in other areas. After laparoscopic splenectomy, the pathology report confirmed a single splenic metastasis from undifferentiated carcinoma of the nasopharyngeal type. The postoperative period was uneventful. We also performed a systematic review of the literature using MEDLINE and Google Scholar databases. All articles reporting cases of splenic metastases from nasopharyngeal carcinoma, with or without histologic confirmation, were evaluated. The literature search yielded 15 relevant articles, which were very heterogeneous in their aims and methods and described only 25 cases of splenic metastases from nasopharyngeal carcinoma. Conclusion: The present review shows that solitary splenic metastases from nasopharyngeal carcinoma are a rare event, but it should be considered in patients presenting with splenic lesions at imaging and a history of primary or recurrent nasopharyngeal carcinoma. No evidence supports a negative impact of splenectomy in patients with solitary splenic metastasis from nasopharyngeal carcinoma