Constitutive stilbene contents of grapevine cluster stems as potential source of resveratrol in wine

Ripe clusters of 8 Vitis vinifera L. varieties (Cabernet franc, Gewürztraminer, Marzemino, Merlot, Moscato, Pinot gris, Sauvignon, Tocai friulano) were analysed to determine the constitutional levels of trans- and cis-resveratrol, ε-viniferin and pterostilbene. Grape stems were extracted by an hydroalcoholic solution, in order to simulate the resveratrol extraction from stems that might occur during alcoholic fermentation, due to the presence of little pieces of stems in the fermenting must. The average stem concentrations of constitutive trans-, cis-resveratrol and ε-viniferin were 142, 0.30, and 70 mu g.g-1 FW, respectively; pterostilbene was not found. Varieties differed significantly in their stilbene content. The percentage of trans-resveratrol extracted from the stems by a hydroalcoholic solution varied from 2.5 % (Gewürztraminer) to 32.9 % (Marzemino), the contribution of the stems to the trans-resveratrol concentration of the hydroalcoholic solution being, on the average, 34 mu g.l-1

Structural analysis of a subduction-related contact in southern Sesia-Lanzo Zone (Austroalpine Domain, Italian Western Alps)

A new foliation trajectory map at 1:10000 scale, represented here with an interpretative structural map, is derived from an original field analysis at 1:5000 scale in the southern Sesia-Lanzo Zone (SLZ). It shows the relative chronology of overprinting foliations, characterised by the mineral assemblages that mark superposed fabrics in each rock type. This map and the associated cross-sections, which synthesise the 3D structural outline of the tectonic contact between the Eclogitic Micaschists Complex (EMC), the Rocca Canavese Thrust Sheets and the Lanzo Ultramafic Complex, allow the correlation of the structural and metamorphic imprints that developed in these crustal and mantle complexes during Alpine subduction. Furthermore, the map and cross-sections allow the immediate perception of the metamorphic environments in which the structural imprints developed in each complex successively under eclogite, blueschist and greenschist facies conditions. The represented structural and metamorphic evolution of the southern end of the SLZ (internal Western Alps) has been inferred based on multiscale structural analysis. The dominant fabrics at the regional scale are two superposed mylonitic foliations that developed under blueschist and greenschist facies conditions, respectively. Metamorphic assemblages underlying the successive fabrics in the different metamorphic complexes allow us to identify contrasting metamorphic evolutions indicating that the tectonic contacts between the EMC, the Rocca Canavese Thrust Sheets and the Lanzo Ultramafic Complex developed under blueschist facies conditions and were successively reactivated during the greenschist facies retrogression

Elicitation and accumulation of stilbene phytoalexins in grapevine berries infected by Botrytis cinerea

At three developmental stages berries of field-grown Castor (interspecific crossing) and Huxelrebe (V. vinifera L. crossing) were in vitro inoculated with two strains of Botrytis cinerea Pers. to investigate the response of berries to fungal infection With respect to the time course of phytoalexin (trans-resveratrol, ε-viniferin and pterostilbene) accumulation and Visual disease symptoms. In infected berries the amounts of ε-viniferin dominated over pterostilbene. The stilbene phytoalexin content decreased during berry development and sugar accumulation. Grape varieties reacted differently to B. cinerea strains with regard to stilbene response and Visual symptoms. Mechanical damage of the berry skin induced uninfected berries to synthesize low amounts of phytoalexins. It can be assumed that after extraction and degradation ε-viniferin of mature berries is a source of resveratrol in wine

Redox agents regulate ion channel activity in vacuoles from higher plant cells

AbstractThe ability of redox agents to modulate certain characteristics of voltage- and calcium-activated channels has been recently investigated in a variety of animal cells. We report here the first evidence that redox agents regulate the activation of ion channels in the tonoplast of higher plants. Using the patch-clamp technique, we have demonstrated that, in tonoplasts from the leaves of the marine seagrass Posidonia oceanica and the root of the sugar beet, a variety of sulphydryl reducing agents, added at the cytoplasmic side of the vacuole, reversibly favoured the activation of the voltage-dependent slow vacuolar (SV) channel. Antioxidants, like dithiothreitol (DTT) and the reduced form of glutathione, gave a reversible increase of the voltage-activated current and faster kinetics of channel activation. Other reducing agents, such as ascorbic acid, also increased the SV currents, although to a lesser extent in comparison with DTT and glutathione, while the oxidising agent chloramine-T irreversibly abolished the activity of the channel. Single channel experiments demonstrated that DTT reversibly increased the open probability of the channel, leaving the conductance unaltered. The regulation of channel activation by glutathione may correlate ion transport with other crucial mechanisms that in plants control turgor regulation, response to oxidative stresses, detoxification and resistance to heavy metals

The nasal delivery of nanoencapsulated statins – An approach for brain delivery

© 2016 Clementino et al. Purpose: Along with their cholesterol-lowering effect, statins have shown a wide range of pleiotropic effects potentially beneficial to neurodegenerative diseases. However, such effects are extremely elusive via the conventional oral administration. The purpose of the present study was to prepare and characterize the physicochemical properties and the in vivo biodistribution of simvastatin-loaded lecithin/chitosan nanoparticles (SVT-LCNs) suitable for nasal administration in view of an improved delivery of the statins to the brain. Materials and methods: Chitosan, lecithin, and different oil excipients were used to prepare nanocapsules loaded with simvastatin. Particle size distribution, surface charge, structure, simvastatin loading and release, and interaction with mucus of nanoparticles were determined. The nanoparticle nasal toxicity was evaluated in vitro using RPMI 2651 nasal cell lines. Finally, in vivo biodistribution was assessed by gamma scintigraphy via Tc99m labeling of the particles. Results: Among the different types of nanoparticles produced, the SVT-LCN_MaiLab showed the most ideal physicochemical characteristics, with small diameter (200 nm), positive surface charge (+48 mV) and high encapsulation efficiency (EE; 98%). Size distribution was further confirmed by nanoparticle tracking analysis and electron microscopy. The particles showed a relatively fast release of simvastatin in vitro (35.6%±4.2% in 6 hours) in simulated nasal fluid. Blank nanoparticles did not show cytotoxicity, evidencing that the formulation is safe for nasal administration, while cytotoxicity of simvastatin-loaded nanoparticles (IC50) was found to be three times lower than the drug solution (9.92 vs 3.50 μM). In rats, a significantly higher radioactivity was evidenced in the brain after nasal delivery of simvastatin-loaded nanoparticles in comparison to the administration of a similar dose of simvastatin suspension. Conclusion: The SVT-LCNs developed presented some of the most desirable characteristics for mucosal delivery, that is, small particle size, positive surface charge, long-term stability, high EE, and mucoadhesion. In addition, they displayed two exciting features: First was their biodegradability by enzymes present in the mucus layer, such as lysozyme. This indicates a new Trojan-horse strategy which may enhance drug release in the proximity of the nasal mucosa. Second was their ability to enhance the nose-to-brain transport as evidenced by preliminary gamma scintigraphy studies

Ultrastable shelled PFC nanobubbles:a platform for ultrasound-assisted diagnostics and therapy

Nanoscale echogenic bubbles (NBs), can be used as a theranostic platform for the localized delivery of encapsulated drugs. However, the generation of NBs is challenging, because they have lifetimes as short as milliseconds in solution. The aim of this work has been the optimization of a preparation method for the generation of stable NBs, characterized by measuring: a) acoustic efficiency, b) nano-size, to ensure passive tumour targeting, c) stability during storage and after injection and d) ability to entrap drugs. NBs are monodisperse and ultrastable, their stability achieved by generation of an amphiphilic multilamellar shell able to efficiently retain the PFC gas. The NBs perform as good acoustic enhancers over a wide frequency range and out of resonant conditions, as tested in both in vitro and in vivo experiments, proving to be a potential platform for the production of versatile carriers to be used in ultrasound-assisted diagnostic, therapeutic and theranostic applications

Report from the HarmoSter study: inter-laboratory comparison of LC-MS/MS measurements of corticosterone, 11-deoxycortisol and cortisone.

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) panels that include glucocorticoid-related steroids are increasingly used to characterize and diagnose adrenal cortical diseases. Limited information is currently available about reproducibility of these measurements among laboratories. The aim of the study was to compare LC-MS/MS measurements of corticosterone, 11-deoxycortisol and cortisone at eight European centers and assess the performance after unification of calibration. Seventy-eight patient samples and commercial calibrators were measured twice by laboratory-specific procedures. Results were obtained according to in-house and external calibration. We evaluated intra-laboratory and inter-laboratory imprecision, regression and agreement against performance specifications derived from 11-deoxycortisol biological variation. Intra-laboratory CVs ranged between 3.3 and 7.7%, 3.3 and 11.8% and 2.7 and 12.8% for corticosterone, 11-deoxycortisol and cortisone, with 1, 4 and 3 laboratories often exceeding the maximum allowable imprecision (MAI), respectively. Median inter-laboratory CVs were 10.0, 10.7 and 6.2%, with 38.5, 50.7 and 2.6% cases exceeding the MAI for corticosterone, 11-deoxycortisol and cortisone, respectively. Median laboratory bias vs. all laboratory-medians ranged from -5.6 to 12.3% for corticosterone, -14.6 to 12.4% for 11-deoxycortisol and -4.0 to 6.5% for cortisone, with few cases exceeding the total allowable error. Modest deviations were found in regression equations among most laboratories. External calibration did not improve 11-deoxycortisol and worsened corticosterone and cortisone inter-laboratory comparability. Method imprecision was variable. Inter-laboratory performance was reasonably good. However, cases with imprecision and total error above the acceptable limits were apparent for corticosterone and 11-deoxycortisol. Variability did not depend on calibration but apparently on imprecision, accuracy and specificity of individual methods. Tools for improving selectivity and accuracy are required to improve harmonization

Organization of networks with tagged nodes and biased links: a priori distinct communities. The case of Intelligent Design Proponents and Darwinian Evolution Defenders

Among topics of opinion formation it is of interest to observe the characteristics of networks with a priori distinct communities. As an illustration, we report on the citation network(s) unfolded in the recent decades through web available works belonging to selected members of the Neocreationist and Intelligent Design Proponents (IDP) and the Darwinian Evolution Defenders (DED) communities. An adjacency matrix of tagged nodes is first constructed; it is not symmetric. A generalization of considerations pertaining to the case of networks with biased links, directed or undirected, is thus presented. The main characteristic coefficients describing the structure of such partially directed networks with tagged nodes are outlined. The structural features are discussed searching for statistical aspects, equivalence or not of subnetworks through the degree distributions, each network assortativity, the global and local clustering coefficients and the Average Overlap Indices. The various closed and open triangles made from nodes, moreover distinguishing the community, are especially listed to calculate the clustering characteristics. The distribution of elements in the rectangular submatrices are specially examined since they represent inter-community connexions. The emphasis being on distinguishing the number of vertices belonging to a given community. Using such informations one can distinguish between opinion leaders, followers and main rivals and briefly interpret their relationships through psychological-like conditions intrinsic to behavior rules in either community. Considerations on other controversy cases with similar social constraints are outlined, as well as suggestions on further, more general, work deduced from our observations on such networks.Comment: 40 pages, 61 references, 7 Tables, 11 Figures, 2 Appendices (giving the adjacency matrices

Small Angle Scattering and Zeta Potential of Liposomes Loaded with Octa(carboranyl)porphyrazine

In this work the physicochemical characterization of liposomes loaded with a newly synthesised carboranyl porphyrazine (H2HECASPz) is described. This molecule represents a potential drug for different anticancer therapies, such as Boron Neutron Capture Therapy, Photodynamic Therapy and Photothermal Therapy. Different loading methods and different lipid mixtures were tested. The corresponding loaded vectors were studied by Small Angle Scattering (SANS and SAXS), light scattering and zeta potential. The combined analysis of structural data at various length scales and the measurement of the surface charge allowed to obtain a detailed characterization of the investigated systems. The mechanisms underlying the onset of differences in relevant physicochemical parameters (size, polydispersity and charge) were also critically discussed