Isolation and Characterization of Wheat Derived Nonspecific Lipid Transfer Protein 2 (nsLTP2).

Numerous studies support the protective role of bioactive peptides against cardiovascular diseases. Cereals represent the primary source of carbohydrates, but they also contain substantial amounts of proteins, therefore representing a potential dietary source of bioactive peptides with nutraceutical activities. The analysis of wheat extracts purified by chromatographic techniques by means of HPLC-UV/nanoLC-nanoESI-QTOF allowed the identification of a signal of about 7 kDa which, following data base searches, was ascribed to a nonspecific lipid-transfer protein (nsLTP) type 2 from Triticum aestivum (sequence coverage of 92%). For the first time nsLTP2 biological activities have been investigated. In particular, in experiments with human umbilical vein endothelial cells (HUVEC), nsLTP2 displayed antioxidant and cytoprotective activities, being able to significantly decrease reactive oxygen species (ROS) levels and to reduce lactate dehydrogenase (LDH) release, generated following oxidative (hydrogen peroxide) and inflammatory (tumor necrosis factor α, interleukin-1ß, and lipopolysaccharide) stimulation. The obtained promising results suggest potential protective role of nsLTP2 in vascular diseases prevention. PRACTICAL APPLICATION: nsLTP 2 peptide is resistant to proteases throughout the gastrointestinal tract and exerts antioxidant and cytoprotective activities. These characteristics could be exploited in vascular diseases prevention.

Transcriptome Profiling of Wheat Seedlings following Treatment with Ultrahigh Diluted Arsenic Trioxide.

Plant systems are useful research tools to address basic questions in homeopathy as they make it possible to overcome some of the drawbacks encountered in clinical trials (placebo effect, ethical issues, duration of the experiment, and high costs). The objective of the present study was to test the hypothesis whether 7-day-old wheat seedlings, grown from seeds either poisoned with a sublethal dose of As2O3 or unpoisoned, showed different significant gene expression profiles after the application of ultrahigh diluted As2O3 (beyond Avogadro's limit) compared to water (control). The results provided evidence for a strong gene modulating effect of ultrahigh diluted As2O3 in seedlings grown from poisoned seeds: a massive reduction of gene expression levels to values comparable to those of the control group was observed for several functional classes of genes. A plausible hypothesis is that ultrahigh diluted As2O3 treatment induced a reequilibration of those genes that were upregulated during the oxidative stress by bringing the expression levels closer to the basal levels normally occurring in the control plants.

Droplet evaporation method as a new potential approach for highlighting the effectiveness of ultra high dilutions.

OBJECTIVE: This study sought to verify whether the droplet evaporation method (DEM) can be applied to assess the effectiveness of ultra-high dilutions (UHDs). We studied the shape characteristics of the polycrystalline structures formed during droplet evaporation of wheat seed leakages. METHODS: The experimental protocol tested both unstressed seeds and seeds stressed with arsenic trioxide 5mM, treated with either ultra-high dilutions of the same stressor substance, or with water as a control. The experimental groups were analyzed by DEM and in vitro growth tests. DEM patterns were evaluated for their local connected fractal dimension (measure of complexity) and fluctuating asymmetry (measure of symmetry exactness). RESULTS: Treatment with arsenic at UHD of both stressed and non-stressed seeds increased the local connected fractal dimension levels and bilateral symmetry exactness values in the polycrystalline structures, as compared to the water treatment. The results of in vitro growth tests revealed a stimulating effect of arsenic at UHD vs. control, and a correlation between the changes in growth rate and the crystallographic values of the polycrystalline structures was observed. CONCLUSIONS: The results indicate that polycrystalline structures are sensitive to UHDs, and so for the first time provide grounds for the use of DEM as a new tool for testing UHD effectiveness. DEM could find application as a treatment pre-selection tool, or to monitor sample conditions during treatment. Moreover, when applied to biological liquids (such as saliva, blood, blood serum, etc.), DEM might provide information about UHD effectiveness on human and animal health.

Phytochemical profile and nutraceutical value of old and modern common wheat cultivars.

Among health-promoting phytochemicals in whole grains, phenolic compounds have gained attention as they have strong antioxidant properties and can protect against many degenerative diseases. Aim of this study was to profile grain phenolic extracts of one modern and five old common wheat (Triticum aestivum L.) varieties and to evaluate their potential antiproliferative or cytoprotective effect in different cell culture systems.Wheat extracts were characterized in terms of antioxidant activity and phenolic composition (HPLC/ESI-TOF-MS profile, polyphenol and flavonoid contents). Results showed that antioxidant activity (FRAP and DPPH) is mostly influenced by flavonoid (both bound and free) content and by the ratio flavonoids/polyphenols. Using a leukemic cell line, HL60, and primary cultures of neonatal rat cardiomyocytes, the potential antiproliferative or cytoprotective effects of different wheat genotypes were evaluated in terms of intracellular reactive oxygen species levels and cell viability. All tested wheat phenolic extracts exerted dose-dependent cytoprotective and antiproliferative effects on cardiomyocytes and HL60 cells, respectively. Due to the peculiar phenolic pattern of each wheat variety, a significant genotype effect was highlighted. On the whole, the most relevant scavenging effect was found for the old variety Verna. No significant differences in terms of anti-proliferative activities among wheat genotypes was observed.Results reported in this study evidenced a correspondence between the in vitro antioxidant activity and potential healthy properties of different extracts. This suggests that an increased intake of wheat grain derived products could represent an effective strategy to achieve both chemoprevention and protection against oxidative stress related diseases.

Development and application of a liquid chromatography-mass spectrometry method to evaluate the glyphosate and aminomethylphosphonic acid dissipation in maize plants after foliar treatment.

A simple and fast method has been developed and validated to measure glyphosate (GLYP) and aminomethylphosphonic acid (AMPA), which were previously derivatized with 9-fluorenylmethylchloroformate (FMOC-Cl), in maize plants using liquid chromatography (LC) coupled to fluorescence (FLD) and electrospray ionization mass spectrometry (ESI-MS) detection. The method has shown to be consistent, reliable, precise, and efficient. Moreover, the limits of detection (LOD) and quantification (LOQ) reached with the proposed method for GLYP and AMPA are lower than the established maximum residue levels (MRLs). The validated method was applied to quantify GLYP and AMPA in genetically modified (GM) maize foliar treated with the herbicide. It has been found that the GLYP dissipation was mainly due to the progressive dilution effect after herbicide treatment. Finally, it was also observed that the GLYP residue dissipation trend in maize shoot (leaves and stem) tissue determined by LC-ESI-MS matched that determined by liquid scintillation.

Profiles of phenolic compounds in modern and old common wheat varieties determined by liquid chromatography coupled with time-of-flight mass spectrometry.

The health-promoting properties of common wheat (Triticum aestivum L.) have been largely attributed to the presence of unique phytochemicals of whole grains. The aim of this study was to profile the phenolic content of 16 old and 6 modern Italian wheat varieties, cropped in the same location and growing season. High variability was observed among the investigated wheat genotypes, both in the free and bound phenolic extracts. The total polyphenol content ranged from 885.5 to 1715.9 µmol GAE/100 g of grain and, on average, the bound fraction contributed for 72.0% to the total phenolic content. As regards the flavonoid content, the free fraction ranged from 50.7 to 106.1 µmol CE/100 g of grain and the bound fraction from 78.3 to 148.9 µmol CE/100 g of grain. Moreover, the interpretation of the mass spectra allowed the characterization of 34 phenolic compounds (104 including isomer forms) belonging to the phenolic acid, flavonoid, coumarin, stilbene, proanthocyanidin and lignan chemical classes. HPLC-ESI-TOF-MS analysis highlighted remarkable differences in the phytochemical fingerprints of old and modern wheat varieties. Six ancient wheat genotypes (Bianco Nostrale, Frassineto, Gentil Rosso, Gentil Rosso Mutico, Marzuolo d'Aqui, Verna) showed phenolic profiles with a number of total compounds and isomer forms much higher than that identified in the modern cultivars. The present findings confirm that ancient wheat may represent a valuable source of biodiversity, especially as regards phenolic compounds. The investigated old wheat genotypes may be successfully used in breeding programs for developing bread wheat varieties with added value in terms of health-promoting phytochemicals.

The influence of tuber mineral element composition as a function of geographical location on acrylamide formation in different Italian potato genotypes.

BACKGROUND: The present study was aimed at examining the effect of tuber mineral composition, distinctive for geographical location, on the expression of acrylamide precursors in three potato genotypes (Arinda, Rossa di Cetica and Sieglinde) in three Italian potato cultivation regions (Puglia, Sicily and Tuscany). RESULTS: Sucrose and amino acids were not correlated with acrylamide formation. In contrast, reducing sugars, limiting with respect to the principal amino acid precursor asparagine, were positively correlated with acrylamide. From analysis of variance, both acrylamide and reducing sugars were not significant for variety but highly significant for location, with higher levels occurring in all three varieties cultivated in Tuscany, followed by Sicily and Puglia respectively. Reducing sugars were negatively correlated with K and Ca and positively correlated with Zn and Cu. Neither N nor P was correlated with reducing sugar content. Path analysis, a statistical technique distinguishing causation and correlation between variables, was implemented to provide additional insight on the interactions between mineral elements and reducing sugars under open field conditions. The variation in reducing sugars in all three varieties was shown to be attributable to Zn. CONCLUSION: Cultivation location has a significant impact on the composition of tuber mineral elements and, as a result, on genotype in the expression of reducing sugars. The negative correlation between Zn and K and the variation in reducing sugar content attributed to the element Zn, which is particularly available in acid soils, are important factors warranting future research aimed at reducing acrylamide formation from an agronomic perspective.