Prolamin content and grain weight in RNAi silenced wheat lines under different conditions of temperature and nitrogen availability

Temperature and nitrogen (N) availability are two important environmental factors that may produce important changes in grain composition during grain filling of bread wheat. In this study, four wheat lines with the down-regulation of gliadins by means of RNA interference (RNAi) have been characterized to determine the effect of thermal stress and N availability on grain weight and quality; with focus on gliadin and glutenin protein fractions. Grain weight was reduced with heat stress (HS) in all RNAi lines, whereas gliadin content was increased in the wild-types. With respect to gliadin content, RNAi lines responded to HS and N availability differently from their respective wild-types, except for ω-gliadin content, indicating a very clear stability of silencing under different environmental conditions. In a context of increased temperature and HS events, and in environments with different N availability, the RNAi lines with down-regulated gliadins seem well suited for the production of wheat grain with low gliadin content.The Spanish Ministry of Economy and Competitiveness (Project AGL2016-80566-P) and the European Regional Development Fund (FEDER) supported this research

Influence of flower head order on phenolic content and quality of globe artichoke at harvest and during twenty-one days of cold storage

Artichoke is one of the vegetables with higher content in phenolic compounds, which are responsible for their taste, flavor and health beneficial effects. However, phenolic profile and concentration depends on many factors, such as genotype, harvest date, and environmental and agronomical conditions. The main aim of this study was to perform a phytochemical characterization of artichoke heads, based on their position on plant (main, secondary and tertiary head) and harvest date, during a complete growing season. Results showed that total identified polyphenol concentration was higher in tertiary heads than secondary and main heads, due to their higher concentration in hydroxycinnamic acid and luteolin derivatives. On the other hand, two postharvest storage experiments with main, secondary and tertiary artichoke heads, harvested in winter and spring, were performed. In addition, tertiary head showed the lowest weight, firmness losses and respiration rate during cold storage which could be attributed to their higher antioxidant compounds. In conclusion, tertiary heads have a greater aptitude to be stored at low temperature from harvesting to consumption since they maintained the quality properties for longer period of time and had higher content of bioactive compounds. However, main artichokes are the most appreciated by consumers due to their larger siz

Characterization of celiac disease related oat proteins: bases for the development of high quality oat varieties suitable for celiac patients

Some studies have suggested that the immunogenicity of oats depends on the cultivar. RP-HPLC has been proposed as a useful technique to select varieties of oats with reduced immunogenicity. The aim of this study was to identify both the avenin protein patterns associated with low gluten content and the available variability for the development of new non-toxic oat cultivars. The peaks of alcohol-soluble avenins of a collection of landraces and cultivars of oats have been characterized based on the RP-HPLC elution times. The immunotoxicity of oat varieties for patients with celiac disease (CD) has been tested using a competitive ELISA based on G12 monoclonal antibody. The oat lines show, on average, seven avenin peaks giving profiles with certain similarities. Based on this similarity, most of the accessions have been grouped into avenin patterns. The variability of RP-HPLC profiles of the collection is great, but not sufficient to uniquely identify the different varieties of the set. Overall, the immunogenicity of the collection is less than 20 ppm. However, there is a different distribution of toxicity ranges between the different peak patterns. We conclude that the RP-HPLC technique is useful to establish groups of varieties differing in degree of toxicity for CD patients.España Ministerio de Economía y Competitividad Project IPT-2011-1321-010000España, Junta de Andalucía Project P12-AGR-176

The Dietary Intervention of Transgenic Low-Gliadin Wheat Bread in Patients with Non-Celiac Gluten Sensitivity (NCGS) Showed No Differences with Gluten Free Diet (GFD) but Provides Better Gut Microbiota Profile

The study evaluated the symptoms, acceptance, and digestibility of bread made from transgenic low-gliadin wheat, in comparison with gluten free bread, in Non-coeliac gluten sensitivity (NCGS) patients, considering clinical/sensory parameters and gut microbiota composition. This study was performed in two phases of seven days each, comprising a basal phase with gluten free bread and an E82 phase with low-gliadin bread. Gastrointestinal clinical symptoms were evaluated using the Gastrointestinal Symptom Rating Scale (GSRS) questionnaire, and stool samples were collected for gluten immunogenic peptides (GIP) determination and the extraction of gut microbial DNA. For the basal and E82 phases, seven and five patients, respectively, showed undetectable GIPs content. The bacterial 16S rRNA gene V1-V2 hypervariable regions were sequenced using the Illumina MiSeq platform and downstream analysis was done using a Quantitative Insights into Microbial Ecology (QIIME) pipeline. No significant differences in the GSRS questionnaires were observed between the two phases. However, we observed a significantly lower abundance of some gut genera Oscillospira, Dorea, Blautia, Bacteroides, Coprococcus, and Collinsella, and a significantly higher abundance of Roseburia and Faecalibacterium genera during the E82 phase compared with the basal phase. The consumption of low-gliadin bread E82 by NCGS subjects induced potentially positive changes in the gut microbiota composition, increasing the butyrate-producing bacteria and favoring a microbial profile that is suggested to have a key role in the maintenance or improvement of gut permeability.España, MINECO Projects AGL2013-48946-C3-1-R, AGL2013-48946-C and AGL2016-80566-

Magnocellular bias in exogenous attention to biologically salient stimuli as revealed by manipulating their luminosity and color

This is the author’s final version of the article, and that the article has been accepted for publication in Journal of Cognitive NeuroscienceExogenous attention is a set of mechanisms that allow us to detect and reorient toward salient events—such as appetitive or aversive—that appear out of the current focus of attention. The nature of these mechanisms, particularly the involvement of the parvocellular and magnocellular visual processing systems, was explored. Thirty-four participants performed a demanding digit categorization task while salient (spiders or S) and neutral (wheels or W) stimuli were presented as distractors under two figure–ground formats: heterochromatic/isoluminant (exclusively processed by the parvocellular system, Par trials) and isochromatic/heteroluminant (preferentially processed by the magnocellular system, Mag trials). This resulted in four conditions: SPar, SMag, WPar, and WMag. Behavioral (RTs and error rates in the task) and electrophysiological (ERPs) indices of exogenous attention were analyzed. Behavior showed greater attentional capture by SMag than by SPar distractors and enhanced modulation of SMag capture as fear of spiders reported by participants increased. ERPs reflected a sequence from magnocellular dominant (P1p, ≃120 msec) to both magnocellular and parvocellular processing (N2p and P2a, ≃200 msec). Importantly, amplitudes in one N2p subcomponent were greater to SMag than to SPar and WMag distractors, indicating greater magnocellular sensitivity to saliency. Taking together, results support a magnocellular bias in exogenous attention toward distractors of any nature during initial processing, a bias that remains in later stages when biologically salient distractors are presen

Preharvest application of methyl salicylate, acetyl salicylic acid and salicylic acid alleviated disease caused by Botrytis cinerea through stimulation of antioxidant system in table grapes

The main goal of this study was to describe impact of preharvest application of methyl salicylate (MeSA), acetyl salicylic acid (ASA) and salicylic acid (SA) on the reduction of disease caused by Botrytis cinerea in two table grape cultivars (‘Crimson’ and ‘Magenta’). Based on previous studies, MeSA and SA were applied at 0.1 and 0.01mM for both cultivars, while ASA was applied at 1mM in ‘Crimson’ and 0.1mM in ‘Magenta’. At time of harvest, berry maturity-quality attributes, bioactive compounds and antioxidant enzymes were determined. In addition, grapes were artificially inoculated with B. cinerea spores, and the berries were ranked for visual decay incidence after 5 days of inoculation. Salicylates preharvest treatments led to higher total acidity, content of bioactive compounds and activity of antioxidant enzymes in treated than in control berries. The application of salicylate derivatives induced resistance to B. cinerea spoilage, since higher percentage of berries with no symptoms was observed and on the contrary, the highest percentages of berries were obtained in control grapes. All preharvest treatments with SA, ASA and MeSA alleviated postharvest disease caused by B. cinerea probably due to increasing levels of phenolic compounds and activity of antioxidant enzymes, although the best results were obtained with MeSA at 0.1 mM. Also, for this treatment and dose, higher quality properties, such as higher concentrations of ascorbic, succinic and fumaric acids, were observed compared with no treated-grapes

Uvbyβ photometry of active-chromosphere binaries .1. The system TZ Coronae Borealis

Simultaneous uvby and Hβ photometry of thee noneclipsing double-lined spectroscopic binary TZ CrB, an active binary system with almost unevolved components, is presented. A small amplitude distortion wave (0.012 mag in y) has been found with maximum light at phase ∽0.75. No variations in color or the βindex during the orbital cycle have been detected within the precision of the observations. The resulting color indices in the standard uvby system allow the estimation of some basic stellar parameters, such as the average effective temperature ∽6000 K and the stellar radii of the component stars ∽1.1 R_⨀. These results, together with available spectroscopic data, permit a consistent picture for this interesting binary to be obtained

Stimulatory Response of Celiac Disease Peripheral Blood Mononuclear Cells Induced by RNAi Wheat Lines Differing in Grain Protein Composition

Wheat gluten proteins are responsible for the bread-making properties of the dough but also for triggering important gastrointestinal disorders. Celiac disease (CD) affects approximately 1% of the population in Western countries. The only treatment available is the strict avoidance of gluten in the diet. Interference RNA (RNAi) is an excellent approach for the down-regulation of genes coding for immunogenic proteins related to celiac disease, providing an alternative for the development of cereals suitable for CD patients. In the present work, we report a comparative study of the stimulatory capacity of seven low-gluten RNAi lines differing in grain gluten and non-gluten protein composition, relevant for CD and other gluten pathologies. Peripheral blood mononuclear cells (PBMCs) of 35 patients with active CD were included in this study to assess the stimulatory response induced by protein extracts from the RNAi lines. Analysis of the proliferative response and interferon-gamma (INF-γ) release of PBMCs demonstrated impaired stimulation in response to all RNAi lines. The lower response was provided by lines with a very low content of α- and γ-gliadins, and low or almost devoid of DQ2.5 and p31–43 α-gliadin epitopes. The non-gluten protein seems not to play a key role in PBMC stimulation.Spanish Ministry of Economy, Industry and competitiveness AGL2016-80566-PEuropean Regional Development Fund (FEDER

Sensitive analysis of recombinant human erythropoietin glycopeptides by on-line phenylboronic acid solid-phase extraction capillary electrophoresis-mass spectrometry

In this study, several chromatographic sorbents: porous graphitic carbon (PGC), aminopropyl hydrophilic interaction (aminopropyl-HILIC), and phenylboronic acid (PBA) were assessed for the analysis of glycopeptides by on-line solid-phase extraction capillary electrophoresis mass spectrometry (SPE-CEMS). As the PBA sorbent provided the most promising results, a PBA-SPE-CE-MS method was developed for the selective and sensitive preconcentration of glycopeptides from enzymatic digests of glycoproteins. Recombinant human erythropoietin (rhEPO) was selected as the model glycoprotein and subjected to enzymatic digestion with several proteases. The tryptic O126 and N83 glycopeptides from rhEPO were targeted to optimize the methodology. Under the optimized conditions, intraday precision, linearity, limits of detection (LODs), and microcartridge lifetime were evaluated, obtaining improved results compared to that from a previously reported TiO2-SPE-CE-MS method, especially for LODs of N-glycopeptides (up to 500 times lower than by CE-MS and up to 200 times lower than by TiO2-SPE-CE-MS). Moreover, rhEPO Glu-C digests were also analyzed by PBA-SPE-CE-MS to better characterize N24 and N38 glycopeptides. Finally, the established method was used to analyze two rhEPO products (EPOCIM and NeuroEPO plus), demonstrating its applicability in biopharmaceutical analysis. The sensitivity of the proposed PBA-SPE-CEMS method improves the existing CE-MS methodologies for glycopeptide analysis and shows a great potential in glycoprotein analysis to deeply characterize protein glycosites even at low concentrations of the protein digest