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Within the complex pathological picture associated to diabetes, high glucose (HG) has "per se" effects on cells and tissues that involve epigenetic reprogramming of gene expression. In fetal tissues, epigenetic changes occur genome-wide and are believed to induce specific long term effects. Human umbilical vein endothelial cells (HUVEC) obtained at delivery from gestational diabetic women were used to study the transcriptomic effects of chronic hyperglycemia in fetal vascular cells using Affymetrix microarrays. In spite of the small number of samples analyzed (n=6), genes related to insulin sensing and extracellular matrix reorganization were found significantly affected by HG. Quantitative PCR analysis of gene promoters identified a significant differential DNA methylation in TGFB2. Use of Ea.hy926 endothelial cells confirms data on HUVEC. Our study corroborates recent evidences suggesting that epigenetic reprogramming of gene expression occurs with persistent HG and provides a background for future investigations addressing genomic consequences of chronic HG., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Glucosinolates are a class of secondary plant metabolites particularly occurring in Cruciferae with potential health-promoting properties, as their hydrolysis products, isothiocyanates, possess chemopreventive and antioxidant activities. In the present study, we systematically studied the in vitro redox behaviour of 15 glucosinolates, by using a range of analytical methods measuring different activities: (i) radical scavenging activity toward peroxyl and toward ABTS radical (chain-breaking activity); (ii) capacity in modulating the in vitro resistance of human low-density lipoprotein (LDL) catalysed by copper (chelating and chain-breaking activity). Data obtained from different assays were compared and analysed by principal component analysis (PCA). PCA allowed us to identify a big cluster of glucosinolates (10 out 15 tested) that do not possess any antioxidant capacity; while, the other five glucosinolates showed moderate and specific antioxidant capacity. Notably, sinalbin and gluconasturtiin were highly active in scavenging ABTS radical and in protecting LDL from copper-catalysed oxidation, respectively. The overall results of this study indicate that just few glucosinolates can act as antioxidants., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

High-fat meals (HFM) induce metabolic stress, leading to the activation of protective mechanisms, including inflammation and endogenous antioxidant defences. In the present study, we investigated the effects of antioxidant-rich fruit juice drinks on the endogenous antioxidant response induced by HFM. In a double-blind, cross-over design (10 d washout), fourteen overweight volunteers were randomly assigned to one of the following interventions: HFM+500 ml placebo beverage (HFM-PB, free from fruit); HFM+500 ml antioxidant beverage 1 (HFM-AB1; apple, grape, blueberry and pomegranate juices and grape skin, grape seed and green tea extracts); HFM+500 ml antioxidant beverage 2 (HFM-AB2; pineapple, black currant and plum juices). HFM-PB consumption increased the plasma levels of thiols (SH) (4 h, P< 0·001) and uric acid (UA) (2 h, P< 0·01) and total radical-trapping antioxidant parameter (TRAP) (4 h, P< 0·01). Following the consumption of drinks, UA production was significantly reduced with respect to placebo beverage consumption 8 h after HFM-AB2 consumption (P< 0·05). SH levels were reduced 0·5 (P< 0·05), 1 (P< 0·05) and 2 h (P< 0·01) after HFM-AB1 consumption and 2, 4 and 8 h (P< 0·05) after HFM-AB2 consumption. Plasma TRAP (2 h, P< 0·001) and urinary ferric reducing antioxidant power (0-8 h, P< 0·01) were increased by HFM-AB1 consumption, the drink with the highest in vitro antioxidant capacity, but not by HFM-AB2 consumption. In urine, UA levels were significantly increased from basal levels after the consumption of HFM-PB and HFM-AB2. However, neither of the beverages increased the urinary excretion of UA with respect to the placebo beverage. In conclusion, the increase in UA and SH levels induced by HFM as part of an endogenous antioxidant response to postprandial stress can be prevented by the concomitant ingestion of antioxidant-rich fruit juice drinks.

It has been suggested that some fruit-based drinks (FBD) may delay the onset of postprandial stress, which is involved in the pathogenesis of many diseases. The majority of the studies, which have investigated the effects of FBD on postprandial stress, involved a placebo that was a drink with the same content in sugars or carbohydrates of the FBD, but without the bioactive antioxidant compounds. These studies were aimed more at evaluating the effect of the antioxidants rather than the effect of the FBD as a whole. Only 4 studies compared the effect of FBD with water as control and did not support the hypothesis that FBD could inhibit postprandial dysmetabolism, as well as the studies that compared the effect of orange juice and cola. Overall, the results suggest a complex relationship between postprandial dysmetabolism, inflammation, and oxidative stress. Furthermore, the inflammatory and oxidative stress markers need further analytical validation and normal ranges should be established in order to reach a firm conclusion. Finally, caution should be taken in the interpretation of the effect of FBD in postprandial studies and the reviewed results suggest that dietary recommendations should aim to limit rather than increase sugar-sweetened beverages consumption.

The present study was performed to evaluate the distribution of phytochemicals in wheat grains and millstreams and the contribution of these compounds to the antioxidant activity. Bioactive molecules, although related to genotype, were highly affected by the milling procedure applied. Carotenoids were unevenly distributed in the kernel and lutein resulted to be the main carotenoid in all samples considered. The WSB method was confirmed to be a useful tool to detect yellow color intensity of semolina but it was not suitable to get information on the true content of carotenoids of the sample. Water-methanol soluble components showed antioxidant activity which decreased from the outer to the inner parts of the kernel. Chloroform-methanol soluble compounds showed generally negligible or negative antioxidant activity due to the inadequate amount of antioxidants unable to counteract oxidative processes already in progress.

Quantitation of tr-cinnamaldehyde, safrole and myristicin was carried out in 70 samples of cola-flavoured soft drinks purchased in eight European countries with the purpose of assessing the variability in the levels of these substances. Results indicated a limited variability in the content of the three substances: the ratio between the 90th and the 10th percentile concentration amounted to 21, 6 and 13 for tr-cinnamaldehyde, safrole and myristicin, respectively. The uncertainty in the assessment of dietary exposure to these substances due to the variability of their level in cola-flavoured drinks was low. Based on these analytical data and on refined food consumption data, estimates of exposure to safrole associated to cola drink consumption, along with Margin of Exposure (MOE) values, were obtained. For high consumers of cola-flavoured soft drinks in certain age groups, within some European countries, MOE values lower than 10,000 resulted, MOE values of 10,000 or higher having been stated by the EFSA as a quantitative criterion to identify low concern from a public health point of view and low priority for risk management actions. The lowest MOE values, from 1900 to 3000, were observed for children and teen agers in the United Kingdom and Ireland., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Epidemiological or in vitro evidence suggests a potential role for flavonoids as anti-inflammatory agents; we investigated the effect of flavonoids-rich foods or supplements on tumor necrosis factor- alpha (TNF-α) and interleukin-6 (IL-6) in long-term placebo-controlled human intervention trials. From 110 human intervention studies selected (MEDLINE, EMBASE, CHORANE, and FSTA databases), 32 long-term placebo-controlled trials were suitable for meta-analysis. After sensitivity analysis, seven studies imputed of bias were excluded and 25 studies were analyzed (TNF-α, n = 2404; IL-6, n = 2174). Levels of TNF-α decreased after flavonoid consumption in the fixed model only (mean difference (MD) (95% CI): -0.098 (-0.188, -0.009), p = 0.032), but metaregression results showed that neither higher dose, nor a longer duration of intervention were associated with a greater effect size. Subgroup analysis did not reveal any significant effect for quercetin and soy, but other sources (red wine, pomegranate, and tea extracts) showed a significant effect size both in fixed (MD (95% CI): TNF-α -0.449 (-0.619, -0.280), p < 0.001; IL-6 -0.346 (-0.612, -0.079), p = 0.011) and random (MD (95% CI): TNF-α -0.783 (-1.476, -0.090), p = 0.027; IL-6, -0.556 (-1.062, -0.050), p = 0.031) effect models. High-quality placebo-controlled trials are needed in order to identify flavonoids as the active ingredients., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background and Aim: A complex relationship between immune system and metabolic pathway exists and can induce oxidative stress. The objective of this study was to design a new methodology allowing the measurement of oxidative status of leukocytes., Methods and Results: We developed a flow cytometry technique, based on C11-BODIPY 581/591 staining, to evaluate peroxidation in leukocytes. We defined the Peroxidation of Leukocytes Index Ratio (PLIR) as the ratio between the damage after AAPH-induced and PMA-induced peroxidation, using Trolox as standard antioxidant. Sensitivity of the method was assessed by correlating results with plasma antioxidant capacity (TRAP and FRAP), levels of endogenous antioxidants (uric acid and sulfhydryls) and markers of metabolic status (cholesterol, triglycerides, glucose and insulin). PLIR measures the ratio between the resistance to exogenous and endogenous ROS injury, independently from baseline level of oxidation, which was directly correlated with plasma cholesterol on lymphocytes (0.738, p=0.029), monocytes (0.691, p=0.047) and neutrophils (0.690, p=0.047). PLIR of lymphocytes was inversely correlated with uric acid (-0.810, p=0.009) and FRAP (-0.738, p=0.029) levels. On the other hand, PLIR of monocytes was directly correlated with the total scavenger antioxidant capacity attributable to nutritional antioxidants (0.738, p=0.029), calculated as the difference between TRAP and the contribution of uric acid and sulfhydryls to its value., Conclusions: This study reports a feasible and reproducible new flow cytometry assay for assessing the leukocytes redox status. PLIR discriminates between reducing and scavenger activities and is able to appreciate the potentially dangerous effect of uric acid on innate immune response., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Spatial information on vineyard soil properties can be useful in precision viticulture. In this paper a combination of high resolution soil spatial information of soil electrical resistivity (ER) and ancillary topographic attributes, such as elevation and slope, were integrated to assess the spatial variability patterns of vegetative growth and yield of a commercial vineyard (Vitis vinifera L. cv. Tempranillo) located in the wine-producing region of La Rioja, Spain. High resolution continuous geoelectrical mapping was accomplished by an Automatic Resistivity Profiler (ARP) on-the-go sensor with an on-board GPS system; rolling electrodes enabled ER to be measured for a depth of investigation approximately up to 0.5, 1 and 2 m. Regression analysis and cluster analysis algorithm were used to jointly process soil resistivity data, landscape attributes and grapevine variables. ER showed a structured variability that matched well with trunk circumference spatial pattern and yield. Based on resistivity and a simple terrain attribute uniform management units were delineated. Once a spatial relationship to target variables is found, the integration of point measurement with continuous soil resistivity mapping is a useful technique to identify within-plots areas of vineyard with similar status.

A pandemic, very aggressive population of Pseudomonas syringae pv. actinidiae is currently causing severe economic losses to kiwifruit crops worldwide. Upon leaf attack, this Gram-negative bacterium systemically reaches the plant shoot in a week period. In this study, combined 2-DE and nanoLC-ESI-LIT-MS/MS procedures were used to describe major proteomic changes in Actinidia chinensis shoot following bacterial inoculation in host leaf. A total of 117 differentially represented protein spots were identified in infected and control shoots. Protein species associated with plant defence, including type-members of the plant basal defence, pathogenesis, oxidative stress and heat shock, or with transport and signalling events, were the most represented category of induced components. Proteins involved in carbohydrate metabolism and photosynthesis were also augmented upon infection. In parallel, a bacterial outer membrane polypeptide component was identified in shoot tissues, whose homologues were already linked to bacterial virulence in other eukaryotes. Semiquantitative RT-PCR analysis confirmed expression data for all selected plant gene products. All these data suggest a general reprogramming of shoot metabolism following pathogen systemic infection, highlighting organ-specific differences within the context of a general similarity with respect to other pathosystems. In addition to present preliminary information on the molecular mechanisms regulating this specific plant-microbe interaction, our results will foster future proteomic studies aimed at characterizing the very early events of host colonization, thus promoting the development of novel bioassays for pathogen detection in kiwifruit material., (Copyright © 2012 Elsevier B.V. All rights reserved.)

We have studied the nucleotide diversity of common bean, Phaseolus vulgaris, which is characterized by two independent domestications in two geographically distinct areas: Mesoamerica and the Andes. This provides an important model, as domestication can be studied as a replicate experiment. We used nucleotide data from five gene fragments characterized by large introns to analyse 214 accessions (102 wild and 112 domesticated). The wild accessions represent a cross-section of the entire geographical distribution of P. vulgaris. A reduction in genetic diversity in both of these gene pools was found, which was three-fold greater in Mesoamerica compared with the Andes. This appears to be a result of a bottleneck that occurred before domestication in the Andes, which strongly impoverished this wild germplasm, leading to the minor effect of the subsequent domestication bottleneck (i.e. sequential bottleneck). These findings show the importance of considering the evolutionary history of crop species as a major factor that influences their current level and structure of genetic diversity. Furthermore, these data highlight a single domestication event within each gene pool. Although the findings should be interpreted with caution, this evidence indicates the Oaxaca valley in Mesoamerica, and southern Bolivia and northern Argentina in South America, as the origins of common bean domestication., (© 2012 The Authors. New Phytologist © 2012 New Phytologist Trust.)

The involvement of both oxidative stress and hyperlipaemia in atherosclerosis development is well established. Oxidative burst is an innate immune response to infection, the latter being associated also with marked changes in lipid and lipoprotein metabolism, aimed to neutralize endotoxin toxic effects. On the other hand, lipid overload may increase lipopolysaccharide circulating levels and oxidative stress. Whilst these changes may be beneficial from the perspective of host defense, if they become chronic, they likely increase the risk of atherosclerosis. In particular, oxidation of lipoproteins, resulting from an imbalance of the pro- and antioxidant equilibrium, is involved in the pathologic process of atherosclerosis, changing cellular functions. Lipid oxidation, induced by leukocytes derived reactive oxygen species, can amplify foam cell formation through oxidized low density lipoproteins LDL (oxLDL) formation and uptake. The main enzymes, operating during oxidative burst, involved in LDL oxidation are NADPH oxidase and myeloperoxidase. In vitro studies have shown that oxLDL are able to induce many proatherogenic processes, including modulation of oxidative burst. OxLDL may also induce maturation of dendritic cells and regulate the shift from classical (M1) to alternative (M2) macrophage activation and from T helper 1 to T helper 2 response, suggesting that these could act as a bridge between innate and adaptative immunity, both involved in plaque development. Understanding the relationship between oxLDL and leukocyte oxidative burst helps to explain the involvement of innate immune responses in the early phases of atherosclerosis. The present review focuses on this interplay.

The freeze-dried sprouts' juice of Raphanus sativus (L.) cv. Sango was prepared and analysed for the first time. HPLC analysis of total isothiocyanates, after protein displacement, resulted in 77.8 ± 3.0 μmol/g of dry juice while GC-MS analysis of hexane and acetone extracts showed E- and Z-raphasatin (8.9 and 0.11 μmol/g, respectively) and sulforaphene (11.7 μmol/g), summing up to 20.7 ± 1.7 μmol/g of free isothiocyanates. Sprouts' juice contained an unprecedented wealth of anthocyanins and a new fractionation methodology allowed us to isolate 34 mg/g of acylated anthocyanins (28.3 ± 1.9 μmol/g), belonging selectively to the cyanidin family. Analysis was performed by HPLC-PDA-ESI-MS(n) and extended to deacylated anthocyanins and aglycones, obtained, respectively, by alkaline and acid hydrolysis. This study identified 70 anthocyanins, 19 of which have never been described before and 32 of which are reported here in R. sativus for the first time. Sango radish sprouts are exceptional dietary sources of heath-promoting micronutrients., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

In the last years the need to numerically define color by its coordinates in n-dimensional space has increased strongly. Colorimetric calibration is fundamental in food processing and other biological disciplines to quantitatively compare samples' color during workflow with many devices. Several software programmes are available to perform standardized colorimetric procedures, but they are often too imprecise for scientific purposes. In this study, we applied the Thin-Plate Spline interpolation algorithm to calibrate colours in sRGB space (the corresponding Matlab code is reported in the Appendix). This was compared with other two approaches. The first is based on a commercial calibration system (ProfileMaker) and the second on a Partial Least Square analysis. Moreover, to explore device variability and resolution two different cameras were adopted and for each sensor, three consecutive pictures were acquired under four different light conditions. According to our results, the Thin-Plate Spline approach reported a very high efficiency of calibration allowing the possibility to create a revolution in the in-field applicative context of colour quantification not only in food sciences, but also in other biological disciplines. These results are of great importance for scientific color evaluation when lighting conditions are not controlled. Moreover, it allows the use of low cost instruments while still returning scientifically sound quantitative data.

Background: Fusarium oxysporum f. sp. melonis Snyd. & Hans. (FOM) causes Fusarium wilt, the most important infectious disease of melon (Cucumis melo L.). The four known races of this pathogen can be distinguished only by infection on appropriate cultivars. No molecular tools are available that can discriminate among the races, and the molecular basis of compatibility and disease progression are poorly understood. Resistance to races 1 and 2 is controlled by a single dominant gene, whereas only partial polygenic resistance to race 1,2 has been described. We carried out a large-scale cDNA-AFLP analysis to identify host genes potentially related to resistance and susceptibility as well as fungal genes associated with the infection process. At the same time, a systematic reisolation procedure on infected stems allowed us to monitor fungal colonization in compatible and incompatible host-pathogen combinations., Results: Melon plants (cv. Charentais Fom-2), which are susceptible to race 1,2 and resistant to race 1, were artificially infected with a race 1 strain of FOM or one of two race 1,2 w strains. Host colonization of stems was assessed at 1, 2, 4, 8, 14, 16, 18 and 21 days post inoculation (dpi), and the fungus was reisolated from infected plants. Markedly different colonization patterns were observed in compatible and incompatible host-pathogen combinations. Five time points from the symptomless early stage (2 dpi) to obvious wilting symptoms (21 dpi) were considered for cDNA-AFLP analysis. After successful sequencing of 627 transcript-derived fragments (TDFs) differentially expressed in infected plants, homology searching retrieved 305 melon transcripts, 195 FOM transcripts expressed in planta and 127 orphan TDFs. RNA samples from FOM colonies of the three strains grown in vitro were also included in the analysis to facilitate the detection of in planta-specific transcripts and to identify TDFs differentially expressed among races/strains., Conclusion: Our data suggest that resistance against FOM in melon involves only limited transcriptional changes, and that wilting symptoms could derive, at least partially, from an active plant response.We discuss the pathogen-derived transcripts expressed in planta during the infection process and potentially related to virulence functions, as well as transcripts that are differentially expressed between the two FOM races grown in vitro. These transcripts provide candidate sequences that can be further tested for their ability to distinguish between races.Sequence data from this article have been deposited in GenBank, Accession Numbers: HO867279-HO867981.

The aim of this study was to investigate the suitability of active infrared thermography and thermometry in combination with multivariate statistical partial least squares analysis as rapid soil water content detection techniques both in the laboratory and the field. Such techniques allow fast soil water content measurements helpful in both agricultural and environmental fields. These techniques, based on the theory of heat dissipation, were tested by directly measuring temperature dynamic variation of samples after heating. For the assessment of temperature dynamic variations data were collected during three intervals (3, 6 and 10 s). To account for the presence of specific heats differences between water and soil, the analyses were regulated using slopes to linearly describe their trends. For all analyses, the best model was achieved for a 10 s slope. Three different approaches were considered, two in the laboratory and one in the field. The first laboratory-based one was centred on active infrared thermography, considered measurement of temperature variation as independent variable and reported r = 0.74. The second laboratory-based one was focused on active infrared thermometry, added irradiation as independent variable and reported r = 0.76. The in-field experiment was performed by active infrared thermometry, heating bare soil by solar irradiance after exposure due to primary tillage. Some meteorological parameters were inserted as independent variables in the prediction model, which presented r = 0.61. In order to obtain more general and wide estimations in-field a Partial Least Squares Discriminant Analysis on three classes of percentage of soil water content was performed obtaining a high correct classification in the test (88.89%). The prediction error values were lower in the field with respect to laboratory analyses. Both techniques could be used in conjunction with a Geographic Information System for obtaining detailed information on soil heterogeneity.

Copper is an essential natural micronutrient. However, copper used as a plant protection product may have long-term consequences due to its accumulation in the soil. Limitations on copper use have therefore been defined in organic farming (Regulation EC 889/2008). In the light of new developments and evidence, the European Commission has planned to assess whether further restrictions are needed in the quantities of copper permitted. A two-year field trial was therefore set up with new copper formulations to evaluate the possibility of reducing the copper quantities applied with treatments and consequently to reduce copper soil residues. Plots were prepared, each containing 12 plants and repeated four times in randomized blocks. The test organism was Plasmopara viticola (Berk. and M.A. Curtis) Berl. and De Toni. Cupric formulations characterised by a low metallic content (Glutex CU 90 and Labicuper) were tested in comparison with a reference product (standard) and an untreated control. Evaluations of treatments were carried out periodically on 100 leaves and 100 bunches for each replicate. Data obtained were subjected to statistical analysis. Chemical analyses were performed to determine copper residues on leaves, grapes and soil. Samplings of leaves and grapes were carried out for each replicate. Soil samples were taken from 0-20 cm and 20-40 cm depth. Total copper was determined using spectrophotometry in atomic absorption by acetylene-air flame (FAAS at lambda = 324.8 microm). The results showed that the tested products were effective in controlling downy mildew with a lower copper dosage than with the cupric formulations used as a standard. Glutex CU 90 formulation led to an annual input of copper that was a little more than a third compared to the standard and Labicuper about a fifth or a sixth. At harvest, copper levels in grapes were much lower than RML (fixed at 50 mg/kg). With regard to the impact of cupric treatments on organic vineyard soil, no statistically significant differential increase in Cu residue was observed in soil between tested products versus untreated control. In conclusion, the environmental impact of copper in organic viticulture could be minimized through the new cupric formulations developed by agrochemical companies.

High-quality standards in blueberry juice can be obtained only taking into account fruit compositional variability and its preservation along the processing chain. In this work, five highbush blueberry cultivars from the same environmental growing conditions were individually processed into juice after an initial blanching step and the influence was studied of the cultivar on juice phenolic content, distribution and relative antioxidant activity, measured as scavenging capacity on the artificial free-radical 2,2-diphenyl-1-picrylhydrazyl (DPPH*). A chromatographic protocol was developed to separate all main phenolic compounds in berries. A total of 15 glycosylated anthocyanins, catechin, galactoside, glucoside, and rhamnoside quercetin 3-derivatives, and main benzoic and cinnamic acids were identified. The total content and relative distribution in anthocyanins, chlorogenic acid, and quercetin of each juice were dependent upon cultivar, and the total content was highly correlated (rxy=0.97) to the antioxidant capacity. A selective protective effect of berry blanching in juice processing can be observed on more labile anthocyanin compounds.