Potent,Metabolically Stable 2‐Alkyl‐8‐(2H‐1,2,3‐triazol‐2‐yl)‐9H‐adenines as Adenosine A2A Receptor Ligands

Inhibition of adenosine A_2A receptors has been shown to elicit a therapeutic response in preclinical animal models of Parkinson’s disease (PD). We previously identified the triazolo‐9H‐purine, ST1535, as a potent A_2AR antagonist. Studies revealed that ST1535 is extensively hydroxylated at the ω‐1 position of the butyl side chain. Here, we describe the synthesis and evaluation of derivatives in which the ω‐1 position has been substituted (F, Me, OH) in order to block metabolism. The stability of the compounds was evaluated in human liver microsomes (HLM), and the affinity for A_2AR was determined. Two compounds, (2‐(3,3‐dimethylbutyl)‐9‐methyl‐8‐(2H‐1,2,3‐triazol‐2‐yl)‐9H‐purin‐6‐amine ( 3 b ) and 4‐(6‐amino‐9‐methyl‐8‐(2H‐1,2,3‐triazol‐2‐yl)‐9H‐purin‐2‐yl)‐2‐methylbutan‐2‐ol ( 3 c ), exhibited good affinity against A_2AR (K_i=0.4 nM and 2 nM , respectively) and high in vitro metabolic stability (89.5 % and 95.3 % recovery, respectively, after incubation with HLM for two hours).     

Exploitation of Polyphenolic Extracts from Grape Marc as Natural Antioxidants by Encapsulation in Lipid-Based Nanodelivery Systems

Phenolic compounds were extracted from grape marc by means of high pressure and temperature extraction. In order to increase their dispersability in the aqueous phase, the polyphenolic extracts were encapsulated at a final concentration of 0.1 % (w/w) in nanoemulsion-based delivery systems, which were formulated with natural ingredients, using either a liquid (sunflower oil) or a solid (palm oil) lipid phase, as well as the combination of a hydrophilic and hydrophobic emulsifier, and were produced by high-pressure homogenization. The delivery systems were characterized in terms of physicochemical stability under accelerated ageing (storage at 4 °C, 30 °C, and 55 °C for 14 days), by recording the evolution of the mean droplet size, the creaming index as well as the UV–vis absorption spectra of the encapsulated polyphenols. The antioxidant activity of the encapsulated extracts was measured with two different chemical assays (FRAP and ORAC) and a cellular antioxidant assay. Sunflower oil-based nanoemulsions resulted to be the most physically and chemically stable, with no significant variation of the mean droplet size and no degradation of the encapsulated compounds under the different conditions tested. The FRAP and ORAC assays showed that the antioxidant compounds, when encapsulated, are as available as unencapsulated polyphenols in scavenging the peroxyl radicals (ORAC), but are less available in reducing the ferric tripyridyltriazine complexes (FRAP). Remarkably, the cellular antioxidant activity was significantly higher for the encapsulated grape marc polyphenols than for the unencapsulated ones, suggesting the fundamental role of the nanoemulsions in favoring the delivery through the biological membranes.     

A bacteriocin-like substance produced from Lactobacillus pentosus 39 is a natural antagonist for the control of Aeromonas hydrophila and Listeria monocytogenes in fresh salmon fillets

We studied the ability of Lactobacillus pentosus 39, a BLS (Bacteriocin-like substance)-producing strain, to control the growth of Aeromonas hydrophila ATCC 14715 and Listeria monocytogenes ATCC 19117 artificially added to fresh salmon fillets at refrigeration temperatures and under simulated cold-chain break conditions.At refrigeration temperatures, Lb. pentosus 39 protective culture and its putative bacteriocin significantly reduced A. hydrophila counts compared with the control (2.1 and 1.4 log CFU/g reductions, respectively). Similar behaviour was observed for L. monocytogenes (3.6 and 1.3 log CFU/g reductions, respectively).Under simulated cold-chain break conditions, an increase in temperature (30°C for 12h) produced an evident increase in the development of A. hydrophila, L. monocytogenes, but also of Lb. pentosus 39, with a consequent increase in BLS production. This condition resulted in a greater reduction of both pathogens compared with samples stored at 4°C throughout the experiment (2.8 log CFU/g reduction for A. hydrophila, 5.8 log CFU/g reduction for L. monocytogenes). In samples treated with the putative bacteriocin alone, a less marked decrease was observed.Our study demonstrates the capability of Lb. pentosus 39 to control the growth of psychrotrophic bacteria in an experimental seafood model system. A similar biopreservation technology could provide more prolonged shelf-life during storage of ready-to-eat seafood, ensuring safety, even under extreme conditions.     

Single‐Step Deposition of Au‐ and Pt‐Nanoparticle‐Functionalized Tungsten Oxide Nanoneedles Synthesized Via Aerosol‐Assisted CVD,and Used for Fabrication of Selective Gas Microsensor Arrays

Tungsten oxide nanostructures functionalized with gold or platinum NPs are synthesized and integrated, using a single‐step method via aerosol‐assisted chemical vapour deposition, onto micro‐electromechanical system (MEMS)‐based gas‐sensor platforms. This co‐deposition method is demonstrated to be an effective route to incorporate metal nanoparticles (NP) or combinations of metal NPs into nanostructured materials, resulting in an attractive way of tuning functionality in metal oxides (MOX). The results show variations in electronic and sensing properties of tungsten oxide according to the metal NPs introduced, which are used to discriminate effectively analytes (C₂H₅OH, H₂, and CO) that are present in proton‐exchange fuel cells. Improved sensing characteristics, in particular to H₂, are observed at 250 °C with Pt‐functionalized tungsten oxide films, whereas non‐functionalized tungsten oxide films show responses to low concentrations of CO at low temperatures. Differences in the sensing characteristics of these films are attributed to the different reactivities of metal NPs (Au and Pt), and to the degree of electronic interaction at the MOX/metal NP interface. The method presented in this work has advantages over other methods of integrating nanomaterials and devices, of having fewer processing steps, relatively low processing temperature, and no requirement for substrate pre‐treatment.     

Temperature and storage effects on antioxidant activity of juice from red and white grapes

The effect of temperature and storage time on the phenolic composition and on the antioxidant activity of grape (Vitis vinifera L.) juices made using a red (Sangiovese, SG) and a white (Muscat of Alexandria, MA) variety was studied. Total phenolics, flavonoids, flavan‐3‐ols and hydroxycinnamoyl‐tartrates (HCT) were determined on grape extracts (GE) and juices. Total anthocyanins and anthocyanins composition were measured on GE. The antioxidant activity was assessed by means of two different in vitro tests: scavenging of the stable 2,2‐diphenyl‐1‐picrylhydrazyl radical and the inhibition of tyrosine nitration mediated by authentic peroxynitrite (ONOO^?). All the juices were analysed after 24 h and 2 weeks of storage by means of two‐way anova (factors being cultivar and temperature). Anthocyanins were not detected in MA extract, whereas in SG their content was 534 mg malvidin‐equivalent per 100 g of dry matter (d.m.) and malvidin derivatives (glycosylated and coumaroylated) were the most represented (respectively 169 and 41 malvidin‐equivalent per 100 g of d.m.). HCT content of the extracts was higher in SG (overall +33%). Also in grape juices, HCT were lower in MA and in this variety, the trans‐fertaric and cis‐coutaric acids were also undetectable. Cultivar effect proved to be highly significant, while no significant differences in the phenolic composition were observed for storage temperatures (4 and ?20 °C) and cultivar × temperature interaction. However, when statistical analysis was focused on each cultivar, MA was found to be more sensible to storage conditions and a significant reduction in total phenolics (?20%) and flavonoids (?53%) content and in the ONOO^? scavenging potential (?32%) was evident after 2 weeks at 4 °C (when compared with the same storage temperature after 24 h). On the contrary, SG juices did not show significant differences among the four storage treatments investigated. These results could be explained suggesting that anthocyanins presence in red grape plays a key role in juice stability.     

Effect of inulin as a prebiotic to improve growth and counts of a probiotic cocktail in fermented skim milk

Inulin was used as a prebiotic to improve quality of skim milk fermented by pure cultures of Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus bulgaricus and Bifidobacterium lactis, binary co-cultures with Streptococcus thermophilus, or a cocktail containing all them. Inulin supplementation to pure cultures lowered the generation time, with particular concern to S. thermophilus and L. acidophilus. The generation time of all micro-organisms decreased in the following order: mono-cultures, co-cultures, cocktail. It was demonstrated a synergism between S. thermophilus and the other strains and a bifidogenic effect of inulin. Enumerations of L. rhamnosus in cocktail markedly decreased compared to co-cultures likely because of greater competition for the same substrates. The results of this work highlight the industrial potential of the cocktail, mainly in terms of fermentation acceleration.     

Carotenoids from tomatoes inhibit heterocyclic amine formation

Tomatoes are a rich source of antioxidants and they are commonly used for meat cooking in Mediterranean countries. The effects of tomato antioxidants on the formation of carcinogenic/mutagenic heterocyclic amines were investigated. A liquid model system containing as precursors creatinine, glucose and glycine in molar concentrations comparable to those present in bovine meat (chemical model system) was employed. A freeze-dried bovine meat juice (meat juice model system) was also used for some experiments. In both model systems, an inhibiting effect of tomato carotenoid fraction on the formation of imidazoquinolines (IQx, MeIQx and DiMeIQx) was observed. Using carotenoid extract at a concentration of 1000 ppm, inhibitions of 36% and 11% of IQx and MeIQx formation respectively in the chemical system and of 13% of MeIQx and of 5% of 4,8-DiMeIQx in the meat juice system was observed. The effect of the main tomato flavonoid, quercetin, was investigated using the meat juice system. Quercetin gave an inhibition of MeIQx formation between 9% and 57% with a maximum effect of 67% at 10 ppm.     

Image restoration in chirp-pulse microwave CT (CP-MCT)

Chirp-pulse microwave computed tomography (CP-MCT) is a technique for imaging the distribution of temperature variations inside biological tissues. Even if resolution and contrast are adequate to this purpose, a further improvement of image quality is desirable. In this paper, we discuss the blur of CP-MCT images and we propose a method for estimating the corresponding point spread function (PSF). To this purpose we use both a measured and a computed projection of a cylindrical phantom. We find a good agreement between the two cases. Finally the estimated PSF is used for deconvolving data corresponding to various kinds of cylindrical phantoms. We use an iterative nonlinear deconvolution method which assures nonnegative solutions and we demonstrate the improvement of image quality which can be obtained in such a way.     

Reduction of immunoreactivity of bovine beta-lactoglobulin upon combined physical and proteolytic treatment

Bovine beta-lactoglobulin was hydrolyzed with trypsin or chymotrypsin before, during and after treatment at 600 MPa and pH 6.8 for 10 min at 30, 37 and 44 degrees C. The extent of beta-lactoglobulin hydrolysis under pressure was noticeably higher than at atmospheric pressure, particularly when chymotrypsin was used. Addition of proteases at ambient pressure to previously pressure-treated beta-lactoglobulin gave only a modest increase in proteolysis with respect to the untreated protein. Products of enzyme hydrolysis under pressure were separated by reverse-phase HPLC, and were found to be different from those obtained at atmospheric pressure when chymotrypsin was used. The residual immunochemical reactivity of the products of combined pressure-enzyme treatment was assessed on the unresolved hydrolysates by ELISA tests using polyclonal and monoclonal antibodies, and on individual hydrolytic fractions by Western Blotting using sera of paediatric patients allergic to whey proteins in cow milk. The immunoreactivity of the whole hydrolysates was related to their content of residual intact beta-lactoglobulin, and no immunochemical reactivity was found for all the products of chymotrypsin hydrolysis under pressure. The results indicate that chymotrypsin effectively hydrolysed hydrophobic regions of beta-lactoglobulin that were transiently exposed during the pressure treatments and that were not accessible in the native protein or in the protein that had been previously pressure treated.     

Extraction of phenolic compounds from Vitex agnus-castus L.

The primary objective of this study was to valorized Vitex agnus-castus residues in terms of phenolic compounds. The effects of extraction time (30–360 min), solid to liquid ratio (0.1–0.3 g_DryBiomass/ml_Solvent), type of solvent and different tissue types (leave, roots and seeds) on total polyphenols, o-diphenols, total flavonoids and anthocyanins were evaluated. The highest total polyphenol (31.5 mg_{CaffeicAcidEquivalent}/g_DryBiomass) and o-diphenol (12.4 mg_{CaffeicAcidEquivalent}/g_DryBiomass) contents were obtained from methanolic extract of leaves after 180 min using a solid/liquid ratio of 0.1 g_DryBiomass/ml_Solvent, while total flavonoids, reached a maximum value of 19.4 mg_{CatechinEquivalent}/g_DryBiomass after 360 min under the same conditions. Roots of V. agnus-castus were found to be a good source of anthocyanins with the highest yield of 0.62 mg_{MalvidinEquivalent}/g_DryBiomass using ethanol as a solvent (180 min and 0.2 g_DryBiomass/ml_Solvent). The maximum antiradical power (178.5 μl_extract/μg_DPPH) was exhibited by the methanolic leave extract obtained after 360 min at solid/liquid ratio of 0.3 g_DryBiomass/ml_Solvent.