Platelet-Derived Extracellular Vesicles for Regenerative Medicine

Extracellular vesicles (EVs) present a great potential for the development of new treatments in the biomedical field. To be used as therapeutics, many different sources have been used for EVs obtention, while only a few studies have addressed the use of platelet-derived EVs (pEVs). In fact, pEVs have been shown to intervene in different healing responses, thus some studies have evaluated their regenerative capability in wound healing or hemorrhagic shock. Even more, pEVs have proven to induce cellular differentiation, enhancing musculoskeletal or neural regeneration. However, the obtention and characterization of pEVs is widely heterogeneous and differs from the recommendations of the International Society for Extracellular Vesicles. Therefore, in this review, we aim to present the main advances in the therapeutical use of pEVs in the regenerative medicine field while highlighting the isolation and characterization steps followed. The main goal of this review is to portray the studies performed in order to enhance the translation of the pEVs research into feasible therapeutical applications.     

Absorption of anthocyanins through intestinal epithelial cells – Putative involvement of GLUT2

Anthocyanins bioavailability is a major issue regarding their biological effects and remains unclear due to few data available on this matter. This work aimed to evaluate the absorption of anthocyanins at the intestine using Caco‐2 cells. Anthocyanin extract, rich in malvidin‐3‐glucoside, was obtained from red grape skins and tested on Caco‐2 cells. The absorption of anthocyanins, in absence or presence of 1% ethanol, was detected by HPLC/DAD/LC‐MS. Our results showed that this transport was significantly increased in the presence of ethanol especially after 60 min of incubation. In addition, cells that were pretreated for 96 h with anthocyanins (200 μg/mL) showed an increase of their own transport (about 50% increase). Expression of glucose transporters sodium‐dependent glucose transporter 1, facilitative glucose transporters 5, and facilitative glucose transporters 2 was assessed by RT‐PCR. It was found that facilitative glucose transporters 2 expression was increased (60%) in Caco‐2 cells pretreated with anthocyanins, by comparison with controls. When the effect of anthocyanin extract on ³H‐2‐deoxy‐D ‐glucose uptake was tested, an inhibitory effect was observed (about 60% decrease). However, the malvidin aglycone was tested and had no effect. In conclusion, anthocyanins could be absorbed through Caco‐2 cells, and can interfere with their own transport and also with glucose intestinal uptake.     

Determination of oil and water in olive and olive pomace by NIR and multivariate analysis

Fourier transform (FT) Near Infrared Spectroscopy (NIR) spectrometry in combination with partial least squares (PLS) regression was used for direct, reagent-free determination fat and moisture content in milled olive and olive pomace. The two calibration models obtained were built with samples from two years harvest (2006/2007) and have a good predictive power considering the nature of the samples and are both being used in an industrial plant.     

Estimation of olive oil acidity using FT-IR and partial least squares regression

Olive oil characteristics are directly related to olive quality. Information about olive quality is of paramount importance to olive and olive oil producers, in order to establish its price. Real-time characterization of the olives avoids mixtures of high quality with low quality fruits, and allows improvement of olive oil quality. This work describes an indirect determination of olive acidity and that allows a rapid evaluation of olive oil quality. The applied method combines chemical analysis (30 min Soxhlet olive pomace extraction) in tandem with a spectroscopic technique (FT-IR) and multivariate regression (PLS1). The most suitable calibration model found used SNV pre-processing and was built with 4 Latent Variables giving a RMSECV of 8.7% and a Q² of 0.97. This accurate calibration model allows the estimation of olive acidity using a FT-IR spectrum of the corresponding Soxhlet oil dry extract and therefore is a suitable method for indirect determination of FFA in olives.     

Biodiversity of bacteriophages infecting Lactococcus lactis starter cultures

In the current study, we characterized 137 Lactococcus lactis bacteriophages that had been isolated between 1997 and 2012 from whey samples obtained from industrial facilities located in 16 countries. Multiplex PCR grouping of these 137 phage isolates revealed that the majority (61.31%) belonged to the 936 group, with the remainder belonging to the P335 and c2 groups (23.36 and 15.33%, respectively). Restriction profile analysis of phage genomic DNA indicated a high degree of genetic diversity within this phage collection. Furthermore, based on a host-range survey of the phage collection using 113 dairy starter strains, we showed that the c2-group isolates exhibited a broader host range than isolates of the 936 and P335 groups.     

Hazelnut Allergens: Molecular Characterization,Detection, and Clinical Relevance

In last few years, special attention has been given to food-induced allergies, in which hazelnut allergy is highlighted. Hazelnut is one of the most commonly consumed tree nuts, being largely used by the food industry in a variety of processed foods. It has been regarded as a food with potential health benefits, but also as a source of allergens capable of inducing mild to severe allergic reactions in sensitized individuals. Considering the great number of reports addressing hazelnut allergens, with an estimated increasing trend, this review intends to assemble all the relevant information available so far on the following main issues: prevalence of tree nut allergy, clinical threshold levels, molecular characterization of hazelnut allergens (Cor a 1, Cor a 2, Cor a 8, Cor a 9, Cor a 10, Cor a 11, Cor a 12, Cor a 14, and Cor a TLP) and their clinical relevance, and methodologies for detection of hazelnut allergens in foods. A comprehensive overview of the current data about the molecular characterization of hazelnut allergens is presented, relating to biochemical classification and biological function with clinical importance. Recent advances in hazelnut allergen detection methodologies are summarized and compared, including all the novel protein-based and DNA-based approaches.     

Synthesis and Biological Studies of Pyrazolyl‐Diamine PtII Complexes Containing Polyaromatic DNA‐Binding Groups

New [PtCl(pz*NN)]ⁿ⁺ complexes anchored by pyrazolyl‐diamine (pz*NN) ligands incorporating anthracenyl or acridine orange DNA‐binding groups have been synthesized so as to obtain compounds that would display synergistic effects between platination and intercalation of DNA. Study of their interaction with supercoiled DNA indicated that the anthracenyl‐containing complex L²Pt displays a covalent type of binding, whereas the acridine orange counterpart L³Pt shows a combination of intercalative and covalent binding modes with a strong contribution from the former. L²Pt showed a very strong cytotoxic effect on ovarian carcinoma cell lines A2780 and A2780cisR, which are, respectively, sensitive to and resistant to cisplatin. In these cell lines, L²Pt is nine to 27 times more cytotoxic than cisplatin. In the sensitive cell line, L³Pt showed a cytotoxic activity similar to that of cisplatin, but like L²Pt was able significantly to overcome cisplatin cross‐resistance. Cell‐uptake studies showed that L²Pt accumulates preferentially in the cytoplasm, whereas L³Pt reaches the cell nucleus more easily, as clearly visualized by time‐lapse confocal imaging of live A2870 cells. Altogether, these findings seem to indicate that interaction with biological targets other than DNA might be involved in the mechanism of action of L²Pt because this compound, despite having a weaker ability to target the cell nucleus than L³Pt , as well as an inferior DNA affinity, is nevertheless more cytotoxic. Furthermore, ultrastructural studies of A2870 cells exposed to L²Pt and L³Pt revealed that these complexes induce different alterations in cell morphology, thus indicating the involvement of different modes of action in cell death.     

Synergistic effects between κ-carrageenan and locust bean gum on physicochemical properties of edible films made thereof

The development of mixed systems, formed by locust bean gum (LBG), and κ-carrageenan (κ-car) can offer new interesting applications such as the development of edible films with particular properties. κ-car/LBG blend films with different ratios were developed, and their effects on films’ physical properties were assessed. Thermogravimetric analysis (TGA), X-ray diffraction (XRD) patterns, dynamic mechanical analysis (DMA) and Fourier-transform infrared (FTIR) spectroscopy techniques were used to highlight the interactions between the two polysaccharides. The addition of κ-car to LBG improved the barrier properties of the films leading to a decrease of water vapor permeability (WVP). Improved values of elongation-at-break (EB) were registered when the ratio of κ-car/LBG was 80/20 or 40/60 (% w/w). Moreover, the κ-car/LBG blend films enhance the tensile strength (TS) compared to κ-car and LBG films. FTIR results suggested that hydrogen bonds interactions between κ-car and LBG have a great influence in films’ properties e.g. moisture content, WVP. Therefore, different κ-car/LBG ratios can be used to tailor edible films with enhanced barrier and mechanical properties.     

Pistachio nut allergy: An updated overview

Pistachio nut (Pistacia vera) is highly appreciated for its organoleptic characteristics and potential health benefits. However, this tree nut is also responsible for triggering moderate to severe IgE-mediated reactions in allergic individuals. Currently, pistachio nut allergy has gained some special attention, mainly due to its intrinsic relation with cashew nut allergy. Like for other nuts, the prevalence of pistachio nut allergy seems to be increasing at a global scale. Until now, there are five allergenic proteins officially listed for pistachio nut (Pis v 1, Pis v 2, Pis v 3, Pis v 4 and Pis v 5). Relevant data on their biochemical classification has become available, enabling establishing a correlation with the respective clinical symptoms. The establishment of an effective allergen risk assessment is a key issue for the food industry, policy makers and regulatory agencies. Thus, the availability of fast, specific and sensitive methods to detect trace amounts of allergens in processed foods is crucial. In the specific case of pistachio nut, there are some protein- and DNA-based methods for its detection/quantification in foods, which can aid to verify label information. Accordingly, all relevant research advances on this topic were summarised, updated and critically discussed in this review.