Advances in quantification and analysis of the celiac-related immunogenic potential of gluten.

Gluten-free products have emerged in response to the increasing prevalence of gluten-related disorders, namely celiac disease. Therefore, the quantification of gluten in products intended for consumption by individuals who may suffer from these pathologies must be accurate and reproducible, in a way that allows their proper labeling and protects the health of consumers. Immunochemical methods have been the methods of choice for quantifying gluten, and several kits are commercially available. Nevertheless, they still face problems such as the initial extraction of gluten in complex matrices or the use of a standardized reference material to validate the results. Lately, other methodologies relying mostly on mass spectrometry-based techniques have been explored, and that may allow, in addition to quantitative analysis, the characterizationof gluten proteins. On the other hand, although the level of 20 mg/kg of gluten detected by these methods is sufficient for a product to be considered gluten-free, its immunogenic potential for celiac patients has not been clinically validated. In this sense, in vitro and in vivo models, such as the organoid technology applied in gut-on-chip devices and the transgenic humanized mouse models, respectively, are being developed for investigating both the gluten-induced pathogenesis and the treatment of celiac disease. Due to the ubiquitous nature of gluten in the food industry, as well as the increased prevalence of gluten-related disorders, here we intend to summarize the available methods for gluten quantification in food matrices and for the evaluation of its immunogenic potential concerning the development of novel therapies for celiac disease to highlight active research and discuss knowledge gaps and current challenges in this field.

Hydrogen peroxide regulates angiogenesis-related factors in tumor cells.

Tumor angiogenesis is required for tumor development and growth, and is regulated by several factors including ROS. H2O2 is a ROS with an important role in cell signaling, but how H2O2 regulates tumor angiogenesis is still poorly understood. We have xenografted tumor cells with altered levels of H2O2 by catalase overexpression into zebrafish embryos to study redox-induced tumor neovascularization. We found that vascular recruitment and invasion were impaired if catalase was overexpressed. In addition, the overexpression of catalase altered the transcriptional levels of several angiogenesis-related factors in tumor cells, including TIMP-3 and THBS1. These two anti-angiogenic factors were found to be H2O2-regulated by two different mechanisms: TIMP-3 expression in a cell-autonomous manner; and, THBS1 expression that was non-cell-autonomous. Our work shows that intracellular H2O2 regulates the expression of angiogenic factors and the formation of a vessel network. Understanding the molecular mechanisms that govern this multifunctional effect of H2O2 on tumor angiogenesis could be important for the development of more efficient anti-angiogenic therapies.

Are Portuguese Cowpea Genotypes Adapted to Drought? Phenological Development and Grain Quality Evaluation.

Along with population growth, global climate change represents a critical threat to agricultural production, compromising the goal of achieving food and nutrition security for all. It is urgent to create sustainable and resilient agri-food systems capable of feeding the world without debilitating the planet. The Food and Agriculture Organization of the United Nations (FAO) refers to pulses as a superfood, as one of the most nutritious crops with high health benefits. Considered to be low-cost, many can be produced in arid lands and have an extended shelf-life. Their cultivation helps reduce greenhouse gases and increases carbon sequestration, also improving soil fertility. Cowpea, Vigna unguiculata (L.) Walp. is particularly drought tolerant, with a wide diversity of landraces adapted to different environments. Considering the importance of knowing and valuing the genetic variability of this species in Portugal, this study assessed the impact of drought on four landraces of cowpea (L1 to L4) from different regions of the country and a national commercial variety (CV) as a reference. The development and evaluation of morphological characteristics were monitored in response to terminal drought (imposed during the reproductive phase), and its effects were evaluated on the yield and quality of the produced grain, namely the weight of 100 grains, color, protein content, and soluble sugars. Under drought conditions, the landraces L1 and L2 showed early maturation as a strategy to avoid water deficit. Morphological alteration of the aerial part of the plants was evident in all genotypes, with a rapid reduction in the number of leaves and a reduction in the number of flowers and pods by between 44 and 72%. The parameters of grain quality, the weight of 100 grains, color, protein, and soluble sugars did not vary significantly, except for sugars of the raffinose family that is associated with the adaptive mechanisms of plants to drought. The performance and maintenance of the evaluated characteristics reflect the adaptation acquired in the past by exposure to the Mediterranean climate, highlighting the potential agronomic and genetic value, still little exploited, that could contribute to production stability, preserved nutritional value, and food safety under water stress.

Elemental Composition and Implications on Brown Rice Flour Biofortified with Selenium.

Rice (Oryza sativa L.) is one of the most economically and socially important cereals in the world. Several strategies such as biofortification have been developed in a way eco-friendly and sustainable to enhance crop productivity. This study implemented an agronomic itinerary in Ariete and Ceres rice varieties in experimental fields using the foliar application of selenium (Se) to increase rice nutritional value. At strategic phases of the plant's development (at the end of booting, anthesis, and at the milky grain stage), they were sprayed with sodium selenate (Na2SeO4) and sodium selenite (Na2SeO3). In the first foliar application plants were sprayed with 500 g Se·ha-1 and in the remaining two foliar applications were sprayed with 300 g Se·ha-1. The effects of Se in the level of micro and macronutrients in brown grains, the localization of Se in these grains, and the subsequent quality parameters such as colorimetric characteristics and total protein were considered. After grain harvesting, the application of selenite showed the highest enrichment in all grain with levels reaching 17.06 µg g-1 Se and 14.28 µg g-1 Se in Ariete and Ceres varieties, respectively. In the Ceres and Ariete varieties, biofortification significantly affected the K and P contents. Regarding Ca, a clear trend prevailed suggesting that Se antagonizes the uptake of it, while for the remaining elements in general (except Mn) no significant differences were noted. Protein content increased with selenite treatment in the Ariete variety but not in Ceres. Therefore, it was possible to conclude, without compromising quality, that there was an increase in the nutritional content of Se in brown rice grain.

Grain Composition and Quality in Portuguese Triticum aestivum Germplasm Subjected to Heat Stress after Anthesis.

Bread wheat (Triticum aestivum) is a major crop worldwide, and it is highly susceptible to heat. In this work, grain production and composition were evaluated in Portuguese T. aestivum germplasm (landraces and commercial varieties), which was subjected to heat after anthesis (grain filling stage). Heat increased the test weight (TW) in Nabão, Grécia and Restauração, indicating an improved flour-yield potential. Mocho de Espiga Branca (MEB) and Transmontano (T94) showed higher thousand-kernel weight (TKW). Gentil Rosso presented increased soluble sugars, which are yeast substrates in the bread-making process. Ardila stood out for its protein increase under heat. Overall SDS was unaffected by higher temperature, but increased in T94, indicating a better dough elasticity for bread-making purposes. Under heat, lipid content was maintained in most genotypes, being endogenous fatty acids (FAs) key players in fresh bread quality. Lipid unsaturation, evaluated through the double bond index (DBI), also remained unaffected in most genotypes, suggesting a lower flour susceptibility to lipoperoxidation. In Grécia, heat promoted a higher abundance of monounsaturated oleic (C18:1) and polyunsaturated linoleic (C18:2) acids, which are essential fatty acids in the human diet. This work highlighted a great variability in most parameters both under control conditions or in response to heat during grain filling. Cluster analysis of traits revealed a lower susceptibility to heat during grain filling in Ardila, Restauração, and Ruivo, in contrast to MEQ, which seems to be more differentially affected at this stage. Characterization and identification of more favorable features under adverse environments may be relevant for agronomic, industrial, or breeding purposes, in view of a better crop adaptation to changing climate and an improved crop sustainability in agricultural systems more prone to heat stress.

Wheat/Gluten-Related Disorders and Gluten-Free Diet Misconceptions: A Review.

In the last 10,000 years, wheat has become one of the most important cereals in the human diet and today, it is widely consumed in many processed food products. Mostly considered a source of energy, wheat also contains other essential nutrients, including fiber, proteins, and minor components, such as phytochemicals, vitamins, lipids, and minerals, that together promote a healthy diet. Apart from its nutritional properties, wheat has a set of proteins, the gluten, which confer key technical properties, but also trigger severe immune-mediated diseases, such as celiac disease. We are currently witnessing a rise in the number of people adhering to gluten-free diets unwarranted by any medical need. In this dynamic context, this review aims to critically discuss the nutritional components of wheat, highlighting both the health benefits and wheat/gluten-related disorders, in order to address common misconceptions associated with wheat consumption.

Review of Structural Features and Binding Capacity of Polyphenols to Gluten Proteins and Peptides In Vitro: Relevance to Celiac Disease.

Polyphenols have been extensively studied due to their beneficial effects on human health, particularly for the prevention and treatment of diseases related to oxidative stress. Nevertheless, they are also known to have an anti-nutritional effect in relation to protein metabolism. This effect is a consequence of its binding to digestive enzymes and/or protein substrates. Dietary gluten is the main trigger of celiac disease, a common immune-based disease of the small intestine and for which the only treatment available is the adherence to a gluten-free diet. Recent studies have addressed the use of dietary polyphenols to interact with gluten proteins and avoid its downstream deleterious effects, taking the advantage of the anti-nutritive nature of polyphenols by protein sequestering. Flavonoids, coumarins and tannins have shown the ability to form insoluble complexes with gluten proteins. One of the most promising molecules has been epigallocatechin-3-gallate, which through its binding to gliadins, was able to reduce gliadins digestibility and its ability to stimulate monolayer permeability and transepithelial transport of immunodominant peptides in cell models. This review focuses on the structural features and binding capacity of polyphenols to gluten proteins and peptides, and the prospects of developing an adjuvant therapy in celiac disease.

Quantification and Tissue Localization of Selenium in Rice (Oryza sativa L., Poaceae) Grains: A Perspective of Agronomic Biofortification.

In worldwide production, rice is the second-most-grown crop. It is considered a staple food for many populations and, if naturally enriched in Se, has a huge potential to reduce nutrient deficiencies in foodstuff for human consumption. This study aimed to develop an agronomic itinerary for Se biofortification of Oryza sativa L. (Poaceae) and assess potential physicochemical deviations. Trials were implemented in rice paddy field with known soil and water characteristics and two genotypes resulting from genetic breeding (OP1505 and OP1509) were selected for evaluation. Plants were sprayed at booting, anthesis and milky grain phases with two different foliar fertilizers (sodium selenate and sodium selenite) at different concentrations (25, 50, 75 and 100 g Se·ha-1). After grain harvesting, the application of selenate showed 4.9-7.1 fold increases, whereas selenite increased 5.9-8.4-fold in OP1509 and OP1505, respectively. In brown grain, it was found that in the highest treatment selenate or selenite triggered much higher Se accumulation in OP1505 relatively to OP1509, and that no relevant variation was found with selenate or selenite spraying in each genotype. Total protein increased exponentially in OP1505 genotype when selenite was applied, and higher dosage of Se also increased grain weight and total protein content. It was concluded that, through agronomic biofortification, rice grain can be enriched with Se without impairing its quality, thus highlighting its value in general for the industry and consumers with special needs.

Data on intracellular localization of RPSA upon alteration of its redox state.

Ribosomal Protein SA (RPSA), a component of the 40S ribosomal subunit, was identified as a H2O2 target in HeLa cells [1]. In order to analyze the intracellular localization of RPSA in different redox states we overexpressed wild-type RPSA (RPSAwt) or RPSA containing two cysteine to serine residue substitutions at positions 148 and 163 (RPSAmut) in HeLa cells. The transfected cells were exposed to H2O2 or N-acetylcysteine (NAC) and RPSA subcellular localization was assessed by immunofluorescence in permeabilized cells. In addition, co-immunofluorescence for RPSA and Ribosomal Protein S6 (RPS6) was performed in cells overexpressing RPSAwt or RPSAmut. Finally, the ribosomal expression of endogenous RPSA in the presence or absence of H2O2 was analyzed by Western blot. The data presented in this work is related to the research article entitled "Hydrogen peroxide regulates cell adhesion through the redox sensor RPSA" [1].

Hydrogen peroxide regulates cell adhesion through the redox sensor RPSA.

To become metastatic, a tumor cell must acquire new adhesion properties that allow migration into the surrounding connective tissue, transmigration across endothelial cells to reach the blood stream and, at the site of metastasis, adhesion to endothelial cells and transmigration to colonize a new tissue. Hydrogen peroxide (H2O2) is a redox signaling molecule produced in tumor cell microenvironment with high relevance for tumor development. However, the molecular mechanisms regulated by H2O2 in tumor cells are still poorly known. The identification of H2O2-target proteins in tumor cells and the understanding of their role in tumor cell adhesion are essential for the development of novel redox-based therapies for cancer. In this paper, we identified Ribosomal Protein SA (RPSA) as a target of H2O2 and showed that RPSA in the oxidized state accumulates in clusters that contain specific adhesion molecules. Furthermore, we showed that RPSA oxidation improves cell adhesion efficiency to laminin in vitro and promotes cell extravasation in vivo. Our results unravel a new mechanism for H2O2-dependent modulation of cell adhesion properties and identify RPSA as the H2O2 sensor in this process. This work indicates that high levels of RPSA expression might confer a selective advantage to tumor cells in an oxidative environment.

The extracellular matrix modulates H2O2 degradation and redox signaling in endothelial cells.

The molecular processes that are crucial for cell function, such as proliferation, migration and survival, are regulated by hydrogen peroxide (H2O2). Although environmental cues, such as growth factors, regulate redox signaling, it was still unknown whether the ECM, a component of the cell microenvironment, had a function in this process. Here, we showed that the extracellular matrix (ECM) differently regulated H2O2 consumption by endothelial cells and that this effect was not general for all types of cells. The analysis of biophysical properties of the endothelial cell membrane suggested that this modification in H2O2 consumption rates was not due to altered membrane permeability. Instead, we found that the ECM regulated GPx activity, a known H2O2 scavenger. Finally, we showed that the extent of PTEN oxidation was dependent on the ECM, indicating that the ECM was able to modulate H2O2-dependent protein oxidation. Thus, our results unraveled a new mechanism by which the ECM regulates endothelial cell function by altering redox balance. These results pinpoint the ECM as an important component of redox-signaling.