NanoPaint: A Tool for Rapid and Dynamic Imaging of Membrane Structural Plasticity at the Nanoscale

Single‐particle tracking with quantum dots (QDs) constitutes a powerful tool to track the nanoscopic dynamics of individual cell membrane components unveiling their membrane diffusion characteristics. Here, the nano‐resolved population dynamics of QDs is exploited to reconstruct the topography and structural changes of the cell membrane surface with high temporal and spatial resolution. For this proof‐of‐concept study, bright, small, and stable biofunctional QD nanoconstructs are utilized recognizing the endogenous neuronal cannabinoid receptor 1, a highly expressed and fast‐diffusing membrane protein, together with a commercial point‐localization microscope. Rapid QD diffusion on the axonal plasma membrane of cultured hippocampal neurons allows precise reconstruction of the membrane surface in less than 1 min with a spatial resolution of tens of nanometers. Access of the QD nanoconstructs to the synaptic cleft enables rapid 3D topological reconstruction of the entire presynaptic component. Successful reconstruction of membrane nano‐topology and deformation at the second time‐scale is also demonstrated for HEK293 cell filopodia and axons. Named “nanoPaint,” this super‐resolution imaging technique amenable to any endogenous transmembrane target represents a versatile platform to rapidly and accurately reconstruct the cell membrane nano‐topography, thereby enabling the study of the rapid dynamic phenomena involved in neuronal membrane plasticity.     

Cashew Apple Juice as Substrate for Lactic Acid Production

The aim of the present work was to investigate the use of cashew apple juice as a low cost substrate for Lactobacillus casei B-442 cultivation and lactic acid production. Ammonium sulfate was employed as the only exogenous nitrogen source. The effect of cashew apple juice reducing sugars and ammonium sulfate concentration and the fermentation pH and temperature on biomass formation, lactic acid production, and productivity were evaluated. The highest productivity (2.36 g/L.h) was obtained applying 50 g/L of reducing sugar from the cashew apple juice supplemented with 6 g/L of ammonium sulfate. The process yield was about 95% when fermentation was carried out at 37 °C with pH controlled at pH 6.5 using NaOH (120 g/L).     

Probiotic fermented milk with high content of polyphenols: Study of viability and bioaccessibility after simulated digestion

This research evaluated the functional potential of a fermented milk made with Streptococcus thermophilus, a probiotic (Bifidobacterium animalis subsp. lactis) and pomegranate juice. Ferric‐reducing antioxidant potential, Trolox equivalent antioxidant capacity, scavenging effect on 1,1‐diphenyl‐2‐picrylhydrazyl free radical, oxygen radical absorbance capacity and Folin–Ciocalteu assays were used to estimate antioxidant activity and phenolic content. Probiotic survival, release and absorption of polyphenols were evaluated using in vitro gastrointestinal digestion. The concentration of polyphenols increased during digestion from 608 mg to 1417  gallic acid equivalents (GAE) /L. About 14% of polyphenols were found on the dialysate portion. The beverage presented high viability of probiotics with high survival rate after digestion (above 7 log cfu/mL).     

Improvement of frozen banana (Musa cavendishii, cv. Enana) colour by blanching: relationship between browning, phenols and polyphenol oxidase and peroxidase activities

 Browning in banana (Musa cavendishii, cv. Enana) processed products is a result of phenol oxidation catalysed by polyphenol oxidase (PPO) and peroxidase (POD) or of other non-enzymatic reactions (Maillard and Strecker mechanisms). Microwave and steam blanching significantly reduced PPO and POD activities and phenol levels in banana flesh, steam blanching being the most effective method for enzyme inactivation. Freezing/thawing processes produced a significant increase in phenol levels in all samples, due to cellular breakdown. After microwave heating browning processes occurred while steam-treated samples did not exhibit a significant colour change. Extractable PPO and POD activities in all banana samples increased as a consequence of freezing/thawing: steam-blanched slices exhibited lower residual activities. High correlations occurred between phenols and browning (r=0.86) in control samples. Blanched samples (microwave or steam) only exhibited correlations between PPO (r=0.80) and POD (r=0.80) activities and browning. Received: 22 February 1996     

Current and future experimental approaches in the study of grape and wine polyphenols interacting gut microbiota

Interactions between polyphenols and gut microbiota are indeed a major issue of current interest in food science research. Knowledge in this subject is progressing as the experimental procedures and analysis techniques do. The aim of this article is to critically review the more leading-edge approaches that have been applied so far in the study of the interactions between grape/wine polyphenols and gut microbiota. This is the case of in vitro dynamic gastrointestinal simulation models that try to mitigate the limitations of simple static models (batch culture fermentations). More complex approaches include the experimentation with animals (mice, rats, pigs, lambs and chicks) and nutritional intervention studies in humans. Main advantages and limitations as well as the most relevant findings achieved by each approach in the study of how grape/wine polyphenols can modulate the composition and/or functionality of gut microbiota, are detailed. Also, common findings obtained by the three approaches (in vitro, animal models and human nutritional interventions) such as the fact that the Firmicutes/Bacteroidetes ratio tends to decrease after the feed/intake/consumption of grape/wine polyphenols are highlighted. Additionally, a nematode (Caenorhabditis elegans) model, previously used for investigating the mechanisms of processes such as aging, neurodegeneration, oxidative stress and inflammation, is presented as an emerging approach for the study of polyphenols interacting gut microbiota. © 2020 Society of Chemical Industry     

Antimicrobial Activity of Binary and Ternary Mixtures of Vanillin,Citral, and Potassium Sorbate in Laboratory Media and Fruit Purées

The susceptibility of Escherichia coli, Salmonella enteritidis, Saccharomyces cerevisiae, and Zygosaccharomyces bailii to binary and ternary mixtures of potassium sorbate (KS), vanillin (V), and citral (C) was evaluated according to the Berenbaum experimental design, in laboratory media. For some V/C combinations, KS inhibitory concentrations were determined in agarized melon and mango purées by the spiral gradient endpoint (SGE) method. In laboratory media, inhibitory antimicrobial combinations were generally additives. For the yeasts, some synergistic effects were observed. All Berenbaum mixtures which resulted inhibitory in laboratory media were confirmed in the fruit purées. When the SGE method was used, several inhibitory ternary mixtures were found. The lowest inhibitory KS concentrations, estimated for a given V/C combination, corresponded to the bacteria assayed in melon purée. Z. bailii was not inhibited at any condition. Some synergistic antimicrobial combinations (595 ppm V + 251 ppm C + 8 ppm KS in melon and 280 ppm V + 123 ppm C + 8 ppm KS in mango purées) could be useful to achieve a desired inhibitory effect in fruit purées while reducing their concentrations.     

Chitosan biobased and intelligent films: Monitoring pH variations

Many factors can influence the shelf-life product, such as pH, water activity, nutrient levels available oxygen, etc. Variations in food pH could occur due to microorganism growth and chemical reactions that may impact flavour, consistency and shelf-life. The objective of this paper was to obtain and characterize the fast pH-colourimetric indicator device, applying a simple manufacturing technique, using food grade and biodegradable materials. The intelligent film was based on natural compounds as chitosan and anthocyanin (pH-colourimetric indicator). Chitosan intelligent films (C-ATH, 2.0 g/100 g) were obtained incorporating anthocyanin (1.0 g/100 g) in matrix films. Initially, a dark violet colour was observed in chitosan films dried. When immersed in different pH buffers, the colour range of the films varied from pink (in acid pH) to bluish-green (in neutral pH) and to violet (in basic pH). The water solubility and water vapour transmission rate of C-ATH were decreased to 60% and 48%, respectively, as compared to control film (CF, without indicator). The mechanical properties such as tensile strength and rigidity were maintained, and the elongation at break was reduced to 47% compared to CF. The advantages of this system were the simple manufacturing process, biodegradability and usage of natural and safe compounds.     

Antimicrobial effectiveness of bioactive packaging materials from edible chitosan and casein polymers: assessment on carrot, cheese, and salami

Antimicrobial packaging is one of the most promising active packaging systems for controlling spoilage and pathogenic microorganisms. In this work, the intrinsic antimicrobial properties of chitosan (CH) were combined with the excellent thermoplastic and film-forming properties of sodium caseinate (SC) to prepare SC/CH film-forming solutions and films. The antimicrobial effectiveness of SC, CH, and SC/CH coatings on the native microfloras of cheese, salami, and carrots was evaluated. In vitro assays through the test tube assay indicated that the most significant antimicrobial effect was achieved by CH and SC/CH solutions on carrot and cheese native microfloras. SC film-forming solutions did not exert antimicrobial activity on any of the native microflora studied. SC, CH, and SC/CH films stored in controlled environments showed that the retention of the antimicrobial action was observed until 5-d storage, at 65% relative humidity in both temperatures (10 °C and 20 °C). In vivo assays were also performed with SC, CH, and SC/CH applied as coatings or wrappers on the 3 food substrates. CH and SC/CH applied at both immersion and wrapper exerted a significant bactericidal action on mesophilic, psychrotrophic, and yeasts and molds counts, showing the 3 microbial populations analyzed a significant reduction (2.0 to 4.5 log CFU/g). An improvement of the bactericidal properties of the CH/SC blend respect to those of the neat CH film is reported. The ionic interaction between both macromolecules enhances its antimicrobial properties. Practical Application: The continuous consumer interest in high quality and food safety, combined with environmental concerns has stimulated the development and study of biodegradable coatings that avoid the use of synthetic materials. Among them, edible coatings, obtained from generally recognized as safe (GRAS) materials, have the potential to reduce weight loss, respiration rate, and improve food appearance and integrity. They can be used in combination with other food preservation techniques in order to extend the effectiveness of the food preservation chain. Moreover, antimicrobial films and coatings have innovated the concept of active packaging and have been developed to reduce, inhibit, or delay the growth of microorganisms on the surface of food in contact with the package. The use of antimicrobials packaging films to control the growth of microorganisms in food can have a significant impact on shelf-life extension and food safety. In addition, antimicrobial films can be prepared by the combination of inherent antimicrobial materials (that is, CH), with good film-forming protein-based ones (that is, SC). Therefore, the objective of this work is to study the performance of 2 biodegradable and edible biopolymers and their combination as natural packages for selected food products.     

Characterization of a multilayer film activated with Lactobacillus curvatus CRL705 bacteriocins

BACKGROUND: Bacteriocins produced by lactic acid bacteria offer enormous promise for food safety preservation. In this study an active multilayer film obtained by the incorporation of lactocin 705 and lactocin AL705, two bacteriocins produced by Lactobacillus curvatus CRL705 with antimicrobial activity against Lactobacillus plantarum CRL691 and Listeria innocua 7, respectively, was characterized for its potential application in active packaging technology. Film activity performance at different storage conditions, bacteriocins transfer into water and sunflower oil, and film surface properties were evaluated. RESULTS: Film activity against L. innocua 7 was maintained during 2, 4 and 6 weeks at 30, 10 and 5 °C respectively. At 30 and 10 °C, activity loss against L. plantarum CRL691 was observed on the second week of storage and after the fourth week at 5 °C. Results showed no significant difference for active multilayer film contact angle and seal properties compared to the control (without bacteriocins). A decrease in lactocin 705 inhibitory activity after sunflower oil contact was observed, while lactocin AL705 remained unaffected. After water contact, film activity was retained for both bacteriocins. CONCLUSIONS: As demonstrated by antimicrobial activity and physico‐mechanical properties retention, lactocin 705 and AL705 active multilayer film present potential for application in active packaging technology. Copyright © 2011 Society of Chemical Industry     

Isolation of a powerful antioxidant from Olea europaea fruit-mill waste: 3,4-Dihydroxyphenylglycol

Nutritional and antioxidant properties of phenolic compounds are important in relation to human health and palatability of products. 3,4-Dihydroxyphenylglycol (DHPG) is a strong antioxidant found in small amounts in virgin olive oil and table olives, with an antioxidant activity even higher than that of the powerful hydroxytyrosol. The origin of this antioxidant is completely unclear since has never been reported as a free plant metabolite. In this respect possible precursors of DHPG have also been discussed in this study. The presence of soluble compounds that either contain DHPG in their molecular structure or act as substrates for its synthesis has been showed for the first time. The quantities of DHPG recovered in olive drupe tissue by thermal treatment exceed widely the values indicated in the literature, showing the release or formation of additional DHPG from precursors after heating. In addition, DHPG obtained under certain extraction conditions from fresh solid waste of two-phase olive oil extraction systems (alperujo) is its most important phenolic compound. Therefore, the solid olive waste is a good source of this simple monomer phenol. The chemical structure, purity and racemic nature of isolated DHPG were also analysed for the first time by NMR experiments.