Water as a solvent of election for obtaining oleuropein-rich extracts from olive (Olea europaea) leaves

Leaves from Olea europaea represent one of the main by-products of the olive oil industry, containing a plethora of bioactive compounds with several promising activities for human health. An organic solvent-free extraction method was developed for the recovery of olive leaf phenols, which obtained an extract containing oleuropein in high amounts. A comparison of various extraction media is reported, together with the total phenolic content, DPPH (2,2-Diphenyl-1-picrylhydrazyl) content, ORAC (oxygen radical absorbance capacity), and polyphenol oxidase activity of the corresponding extracts. The polyphenol profiles and content of the most representative extracts have also been studied. Extraction solvent and temperature significantly influenced the phenolic content and antioxidant activity of the extracts, with hot water representing the solvent of election for the extraction of bioactive compounds from this matrix. All the extracts obtained showed reasonably high total phenol content (TPC) and good DPPH radical scavenging activity; among them, the water extract is characterized by desirable traits and could be used for many industrial applications and human consumption

Edible films and coatings as carriers of living microorganisms: a new strategy towards biopreservation and healthier foods

Edible films and coatings have been extensively studied in recent years due to their unique properties and advantages over more traditional conservation techniques. Edible films and coatings improve shelf life and food quality, by providing a protective barrier against physical and mechanical damage, and by creating a controlled atmosphere and acting as a semipermeable barrier for gases, vapor, and water. Edible films and coatings are produced using naturally derived materials, such as polysaccharides, proteins, and lipids, or a mixture of these materials. These films and coatings also offer the possibility of incorporating different functional ingredients such as nutraceuticals, antioxidants, antimicrobials, flavoring, and coloring agents. Films and coatings are also able to incorporate living microorganisms. In the last decade, several works reported the incorporation of bacteria to confer probiotic or antimicrobial properties to these films and coatings. The incorporation of probiotic bacteria in films and coatings allows them to reach the consumers gut in adequate amounts to confer health benefits to the host, thus creating an added value to the food product. Also, other microorganisms, either bacteria or yeast, can be incorporated into edible films in a biocontrol approach to extend the shelf life of food products. The incorporation of yeasts in films and coatings has been suggested primarily for the control of the postharvest disease. This work provides a comprehensive review of the use of edible films and coatings for the incorporation of living microorganisms, aiming at the biopreservation and probiotic ability of food products.Ana Guimaraes received support through grant SFRH/BD/ 103245/2014 from the Portuguese Foundation for Science and Technology (FCT). Luís Abrunhosa was supported by grant UMINHO/BPD/51/2015 from project UID/BIO/04469/2013 financed by FCT/MEC (OE). This study was supported by FCT under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and of BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Vectors used in Figure were designed by Freepik.info:eu-repo/semantics/publishedVersio

Bergamot Polyphenols Boost Therapeutic Effects of the Diet on Non-Alcoholic Steatohepatitis (NASH) Induced by “Junk Food”: Evidence for Anti-Inflammatory Activity

Wrong alimentary behaviors and so-called “junk food” are a driving force for the rising incidence of non-alcoholic fatty liver disease (NAFLD) among children and adults. The “junk food” toxicity can be studied in “cafeteria” (CAF) diet animal model. Young rats exposed to CAF diet become obese and rapidly develop NAFLD. We have previously showed that bergamot (Citrus bergamia Risso et Poiteau) flavonoids, in the form of bergamot polyphenol fraction (BPF), effectively prevent CAF diet-induced NAFLD in rats. Here, we addressed if BPF can accelerate therapeutic effects of weight loss induced by a normocaloric standard chow (SC) diet. 21 rats fed with CAF diet for 16 weeks to induce NAFLD with inflammatory features (NASH) were divided into three groups. Two groups were switched to SC diet supplemented or not with BPF (CAF/SC±BPF), while one group continued with CAF diet (CAF/CAF) for 10 weeks. BPF had no effect on SC diet-induced weight loss, but it accelerated hepatic lipid droplets clearance and reduced blood triglycerides. Accordingly, BPF improved insulin sensitivity, but had little effect on leptin levels. Interestingly, the inflammatory parameters were still elevated in CAF/SC livers compared to CAF/CAF group after 10 weeks of dietary intervention, despite over 90% hepatic fat reduction. In contrast, BPF supplementation decreased hepatic inflammation by reducing interleukin 6 (Il6) mRNA expression and increasing anti-inflammatory Il10, which correlated with fewer Kupffer cells and lower inflammatory foci score in CAF/SC+BPF livers compared to CAF/SC group. These data indicate that BPF mediates a specific anti-inflammatory activity in livers recovering from NASH, while it boosts lipid-lowering and anti-diabetic effects of the dietary intervention

Biological control of postharvest phytopathogenic molds promoted by food-isolated yeasts

The mechanisms of action of antagonistic food-isolated yeasts, Wickerhamomyces anomalus, Metschnikowia pulcherrima, Aureobasidium pullulans and Saccharomyces cerevisiae, were evaluated. For each yeast species, different mechanisms involved in biocontrol activity against pathogenic molds, such as iron competition, production of cell wall-degrading enzymes, production of VOCs (Volatile Organic Compounds), and biofilm formation were clarified. The second objective was to determine the efficacy of yeast strains in controlling gray mold caused by Botrytis cinerea on table grapes and strawberries, and green and blue mold decays caused by Penicillium digitatum and Penicillium italicum, respectively, on mandarin fruits. Therefore, subsequent studies investigated the potential use of locust bean gum (LBG) edible coating enriched with antagonistic yeast cells and of a commercial carrier (polyacrylammide hydrogel spheres) emitting antifungal VOCs produced by yeasts. Among tested yeast strains, W. anomalus BS91 showed great antagonistic activity both by direct application of yeast cells and by application of VOCs. This strain is known to produce killer toxins that have been demonstrated to be exoglucanases, coded by the genes WaEXG1 and WaEXG2. To better understand the involvement of these genes in the antagonist activity of W. anomalus, characterization of the gene expression level of EXG1 and EXG2 in different yeast-host-pathogen interactions was carried out. The present study demonstrated the efficacy of the selected yeasts as postharvest antifungal agents both by direct application and incorporation in an edible coating (LBG). Moreover, it has been demonstrated as production of VOCs can play an essential role in the antagonistic activity of BCAs, suggesting a future use of polyacrylammide hydrogel beads as VOCs emitters during the shelf-life

Pomegranate Byproduct Extracts as Ingredients for Producing Experimental Cheese with Enhanced Microbiological, Functional, and Physical Characteristics

Pomegranate peel and mesocarp, considered as wastes of fruit processing, are rich sources of beneficial phytochemicals, including hydrolyzable tannins and flavonoids, with proven antimicrobial and antioxidant activity, which can be employed for improving the overall quality of food products. In the present study, extracts from pomegranate peel (PPW) and mesocarp (PMW) were obtained through a water extraction method and evaluated for in vitro antimicrobial activity and polyphenol content. The two extracts were then added during the cheese-making process in order to create a new functional cheese with improved microbiological and physico-chemical characteristics. Antimicrobial in vitro assays evidenced a substantial efficacy of both extracts against Staphylococcus aureus, which often causes staphylococcal food poisoning outbreaks linked to the consumption of raw milk cheeses and artisanal cheeses. For this reason, a simulated cheese contamination was carried out in order to assess if pomegranate extracts can exert antimicrobial activity towards this pathogen even when incorporated into the cheese matrix. Milk enriched with pomegranate extracts (PPW and PMW) was used to produce two different experimental cheeses, which were then evaluated for yield, polyphenol content, and microbiological as well as physico-chemical traits throughout the refrigerated storage. Despite the low concentration of the extracts, the treated cheeses showed an increase in firmness and a slight decrease in S. aureus counts, of more than one log unit in comparison to the control cheese, for up to 12 d of cold storage. Such results support the reuse of agro-food byproducts, in substitution to chemical food preservatives, as the key to a circular economy

A nonalcoholic fatty liver disease model in human induced pluripotent stem cell-derived hepatocytes, created by endoplasmic reticulum stress-induced steatosis

Hepatic steatosis, a reversible state of metabolic dysregulation, can promote the onset of nonalcoholic steatohepatitis (NASH), and its transition is thought to be critical in disease evolution. The association between endoplasmic reticulum (ER) stress response and hepatocyte metabolism disorders prompted us to characterize ER stress-induced hepatic metabolic dysfunction in human induced pluripotent stem cell-derived hepatocytes (hiPSC-Hep), to explore regulatory pathways and validate a phenotypic in vitro model for progression of liver steatosis. We treated hiPSC-Hep with a ratio of unsaturated and saturated fatty acids in the presence of an inducer of ER stress to synergistically promote triglyceride accumulation and dysregulate lipid metabolism. We monitored lipid accumulation by high-content imaging and measured gene regulation by RNA sequencing and reverse transcription quantitative PCR analyses. Our results show that ER stress potentiated intracellular lipid accumulation by 5-fold in hiPSC-Hep in the absence of apoptosis. Transcriptome pathway analysis identified ER stress pathways as the most significantly dysregulated of all pathways affected. Obeticholic acid dose dependently inhibited lipid accumulation and modulated gene expression downstream of the farnesoid X receptor. We were able to identify modulation of hepatic markers and gene pathways known to be involved in steatosis and nonalcoholic fatty liver disease (NAFLD), in support of a hiPSC-Hep disease model that is relevant to clinical data for human NASH. Our results show that the model can serve as a translational discovery platform for the understanding of molecular pathways involved in NAFLD, and can facilitate the identification of novel therapeutic molecules based on high-throughput screening strategies