Comprehensive molecular level characterization of protein- and polyphenol-rich tara (Caesalpinia spinosa) seed germ flour suggests novel hypothesis about possible accidental hazards.

Tara (Caesalpinia spinosa, Leguminosae) seed germ (TSG), a by-product of tara gum (E417) extraction, has been used as a protein- and polyphenol-rich food ingredient for human and animal nutrition. Nevertheless, TSG is the alleged culprit for a recent foodborne outbreak of even severe acute illnesses that have affected hundreds of individuals in the USA, perhaps triggered by nonprotein amino acids such as baikiain. Herein, the composition of TSG has been characterized at molecular level, with a focus on proteins, phenolics, lipids, and mineral composition. TSG contains 43.4 % (w/w) proteins, tentatively identified for the first time by proteomics, and 14 % lipids, consisting of 83.6 % unsaturated fatty acids, especially linoleic acid. Ash is surprising high (6.5 %) because of an elevated concentration of P, K, Ca, and Mg. The detection of a rare earth element such as gadolinium (Gd, 1.6 mg kg-1), likely sourced from anthropogenic pollution, suggests alternative hypotheses for the origin of TSG hazards.

How dietary advanced glycation end products could facilitate the occurrence of food allergy.

BACKGROUND: Food allergy (FA) is one of the most common chronic conditions in children with an increasing prevalence facilitated by the exposure to environmental factors in predisposed individuals. It has been hypothesized that the increased consumption of ultra-processed foods, containing high levels of dietary advanced glycation end products (AGEs), could facilitate the occurrence of FA. OBJECTIVE: We sought to provide preclinical and clinical evidence on the potential role of AGEs in facilitating the occurrence of FA. METHODS: Human enterocytes, human small intestine organ culture, and PBMCs from children at risk for allergy were used to investigate the direct effect of AGEs on gut barrier, inflammation, TH2 cytokine response, and mitochondrial function. Intake of the 3 most common glycation products in Western diet foods, Nε-(carboxymethyl) lysine, Nε-(1-carboxyethyl) lysin, and Nδ-(5-hydro-5- methyl-4-imidazolone-2-yl)-ornithine (MG-H1), and the accumulation of AGEs in the skin were comparatively investigated in children with FA and in age-matched healthy controls. RESULTS: Human enterocytes exposed to AGEs showed alteration in gut barrier, AGE receptor expression, reactive oxygen species production, and autophagy, with increased transepithelial passage of food antigens. Small intestine organ cultures exposed to AGEs showed an increase of CD25+ cells and proliferating crypt enterocytes. PBMCs exposed to AGEs showed alteration in proliferation rate, AGE receptor activation, release of inflammatory and TH2 cytokines, and mitochondrial metabolism. Significant higher dietary AGE intake and skin accumulation were observed children with FA (n = 42) compared with age-matched healthy controls (n = 66). CONCLUSIONS: These data, supporting a potential role for dietary AGEs in facilitating the occurrence of FA, suggest the importance of limiting exposure to AGEs children as a potential preventive strategy against this common condition.

Insights into the Structure-Capacity of Food Antioxidant Compounds Assessed Using Coulometry.

CDAC (coulometrically determined antioxidant capacity) involves the determination of the antioxidant capacity of individual compounds or their mixtures using constant-current coulometry, with electrogenerated Br2 as the titrant, and biamperometric detection of the endpoint via Br2 excess. CDAC is an accurate, sensitive, rapid, and cheap measurement of the mol electrons (mol e-) transferred in a redox process. In this study, the CDAC of 48 individual antioxidants commonly found in foods has been determined. The molar ratio CDAC (CDACχ, mol e- mol-1) of representative antioxidants is ranked as follows: tannic acid > malvidin-3-O-glucoside ≃ curcumin > quercetin > catechin ≃ ellagic acid > gallic acid > tyrosol > BHT ≃ hydroxytyrosol > chlorogenic acid ≃ ascorbic acid ≃ Trolox®. In many cases, the CDACχ ranking of the flavonoids did not comply with the structural motifs that promote electron or hydrogen atom transfers, known as the Bors criteria. As an accurate esteem of the stoichiometric coefficients for reactions of antioxidants with Br2, the CDACχ provides insights into the structure-activity relationships underlying (electro)chemical reactions. The electrochemical ratio (ER), defined as the antioxidant capacity of individual compounds relative to ascorbic acid, represents a dimensionless nutritional index that can be used to estimate the antioxidant power of any foods on an additive basis.

Pasta fortified with C-glycosides-rich carob (Ceratonia siliqua L.) seed germ flour: Inhibitory activity against carbohydrate digesting enzymes.

Carob (Ceratonia siliqua L.) seed germ flour (SGF) is a by-product resulting from the extractionextraction of locust bean gum (E410), which is a texturing and thickening ingredient used for food, pharmaceutical and cosmetic preparations. SGF is a protein-rich edible matrix and contains relatively high amounts of apigenin 6,8-C-di- and poly-glycosylated derivatives. In this work, we prepared durum wheat pasta containing 5 and 10 % (w/w) of SGF and carried out inhibition assays against type-2 diabetes relevant carbohydrate hydrolysing enzymes, namely porcine pancreatic α-amylase and α-glycosidases from jejunal brush border membranes. Nearly 70-80% of the SGF flavonoids were retained in the pasta after cooking in boiling water. Extracts from cooked pasta fortified with 5 or 10% SGF inhibited either α-amylase by 53% and 74% or α-glycosidases by 62 and 69%, respectively. The release of reducing sugars from starch was delayed in SGF-containing pasta compared to the full-wheat counterpart, as assessed by simulated oral-gastric-duodenal digestion. By effect of starch degradation, the SGF flavonoids were discharged in the water phase of the chyme, supporting a possible inhibitory activity against both duodenal α-amylase and small intestinal α-glycosidases in vivo. SGF is a promising functional ingredient obtained from an industrial by-product for producing cereal-based foods with reduced glycaemic index.

Shotgun proteomics for the identification of yeasts responsible for pink/red discoloration in commercial dairy products.

Pink/red discoloration encompasses a series of relatively common spoilage defects of commercial dairy products. In this study, we used shotgun proteomics to identify the microorganism responsible for the production of intensely red-coloured slimes found on the surface of freshly opened commercial spreadable cheese and yogurt samples. Proteome-wide characterization of microbial proteins allowed to identify 1042 and 687 gene products from Rhodotorula spp. in spreadable cheese and yogurt samples, respectively, while no significant protein scores from other microorganisms were recorded. Subsequent microbiological analyses and sequencing of the 26S rRNA gene region supported the proteomic results demonstrating that the microorganism involved was Rhodotorula mucilaginosa, a carotenoid - producing basidiomycetous that can be potentially pathogenic to humans, especially for immunocompromised individuals. This is the first time that shotgun proteomics has been used to identify a microorganism responsible for spoilage in dairy products, proposing it as a relatively fast, sensitive, and reliable alternative or complement to conventional methods for microbial identification.

Evaluation of Novel Rapid Analytical Methods to Categorize Extra Virgin Olive Oil Based on the Coulometrically Determined Antioxidant Capacity and on the Spectrophotometric Assessment of Phenolic Compounds.

The lack of a practical "fit for the purpose" analytical protocol is the main limitation that has hampered the exploitation of the EFSA analytical health claim on the extra virgin olive oil (EVOO) biophenols, more than ten years since its introduction. In this work, two analytical methods recently developed in our laboratories for categorizing EVOO have been evaluated on a set of 16 samples from Cilento (Campania Region, southern Italy) and compared to other commonly used quality indexes. The Coulometrically Determined Antioxidant Capacity (CDAC) is associated with the component responsible for the health-promoting properties and oxidative stability of EVOO. The Fast Blue BB (FBBB) assay consists of the spectrophotometric (420 nm) determination of biophenols-FBBB diazonium coupling products generated in unfractionated EVOO. The FBBB assay and HPLC-UV reference method provide values highly correlated to each other. Fourteen of sixteen EVOO samples with CDAC > 10 mmol kg-1 and FBBB absorbance > 0.5 had HPLC-determined biophenols > 250 mg kg-1, and therefore eligible for the EFSA health claim. Consistently, two EVOO samples with HPLC-determined biophenols < 250 mg kg-1 had CDAC values and FBBB absorbance below the respective thresholds. CDAC and FBBB assays are proposed individually or in combination as methods to categorize EVOO samples in alternative to HPLC-UV.

Optimized extraction and large-scale proteomics of pig jejunum brush border membranes for use in in vitro digestion models.

Despite the physiological importance of the hydrolases from the intestinal brush border membrane (BBM), a step simulating the intestinal digestion has not been included yet in the harmonized protocols of in vitro digestion, due to commercial unavailability of these enzymes and lack of consensus for the conditions of use. The proper utilize of BBM requires a detailed investigation of their enzymatic composition. BBM vesicles were purified from specimens of pig jejunum optimizing previously described methods and assayed for aminopeptidase N and dipeptidyl peptidase IV activity. Large-scale proteomics was carried out with a bottom-up shotgun approach, also performing a rough quantification with the iBAQ (intensity Based Absolute Quantification). Overall, 1428 proteins were identified and functionally classified by gene ontology enrichment analysis. The predominant enzyme fraction (220 gene products) was represented by hydrolases, including peptidases, glycosidases, and lipases. Aminopeptidase N and sucrase-isomaltase represented 52.9 % and 50.2 % of the peptidase and glycosidase abundance, respectively. In addition to expected transporters and cytoskeletal actin-binding proteins, purified BBM vesicles also contains a complex array of protease inhibitors, here described for the first time, that may modulate the activity of hydrolases. Considering the similarity with the human counterpart, intestinal porcine BBM are suited for simulating the human small intestinal digestion.

Agronomic, Physicochemical, Aromatic and Sensory Characterization of Four Sweet Cherry Accessions of the Campania Region.

Sweet cherries (Prunus avium L.) are greatly appreciated fruits worldwide due to their taste, color, nutritional value, and beneficial health effects. The characterization of autochthonous germplasm allows to identify genotypes that possess superior characteristics compared to standard cultivars. In this work, four accessions of sweet cherry from the Campania region (Limoncella, Mulegnana Riccia, Mulegnana Nera and Montenero) were investigated for their morpho-physiological, qualitative, aromatic, and sensorial traits in comparison with two standard cultivars (Ferrovia and Lapins). A high variability in the pomological traits resulted among the samples. Montenero showed comparable fruit weight and titratable acidity to Ferrovia and Lapins, respectively. The highest total soluble solid content was detected in Mulegnana Riccia. A considerable variability in the skin and pulp color of the cherries was observed, varying from yellow-red in Limoncella to a dark red color in Montenero. Mulegnana Nera showed the highest content of polyphenols, flavonoids, anthocyanins, and ascorbic acid compared to the standard cultivars. Volatile organic compounds profile analysis identified 34 volatile compounds, 12 of which were observed at different concentrations in all the sweet cherry genotypes while the others were genotype-dependent. Conservation and cultivation of autochthonous accessions with suitable nutritional and morpho-physiologic characteristics promotes our agrobiodiversity knowledge and allows to better plan future breeding programs.

Proteomics in bovine semitendinosus muscle to assess emerging strategies based on papain injection and ultrasounds on meat tenderization process.

The impact of papain and/or ultrasound treatments on tenderization of semitendinosus muscle through a proteomic approach was studied. Sixteen bovine muscles were submitted to the following treatments: aging at 3 °C (Control), papain injection (PI), ultrasound (US), PI followed by US (PIUS) and US followed by PI (USPI). pH, myofibrillar fragmentation indices (MFI), soluble collagen, texture profile and changes of myofibrillar proteins were investigated after 2, 24, 48 and 96 h of storage. The highest MFI and soluble collagen content were found in PI, PIUS and USPI samples while control samples showed the lowest values. PI samples showed the lowest WBSF and hardness values until 48 h of storage while at 96 h meat from USPI treatment showed WBSF value comparable to PI treatment. The lowest values of cohesiveness, gumminess and chewiness were found in PI samples during all storage times. Proteomic analysis revealed a different quantity and expression of proteins among tenderization treatments. US treatment did not exhibit a significant ability to degrade muscle proteins, while, all treatments containing papain, showed a greater ability to hydrolyse and degrade myofibrillar proteins. PI promoted intense proteolysis leading to an early tenderization process; on the contrary, in PIUS and USPI treatments the sequence of treatments was relevant on meat tenderization. Particularly, USPI treatment, after 96 h, reached the same improvement in tenderness of enzymatic treatment but with slower hydrolysing rate; this could be determinant to preserve textural structure.

Monitoring antioxidants by coulometry: Quantitative assessment of the strikingly high antioxidant capacity of bergamot (Citrus bergamia R.) by-products.

A recently optimized rapid, cheap, and accurate coulometric method has been exploited to determine the antioxidant capacity of bergamot (Citrus bergamia Risso) by-products, including first (FPJ) and second press juices (SPJ), in comparison to analogous products from several citrus species. Extracts from the entire edible part (i.e., juice and pulp) and de-oiled peel of bergamot were also assayed. The Coulometrically Determined Antioxidant Capacity (CDAC) data, expressed as moles of electrons per mass of sample, were evaluated with other parameters such as total phenolic compounds, ascorbic acid, total carotenoids, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical inhibition. The CDAC of bergamot FPJ (39 mmol e- kg-1) was comparable with other citrus juices (20-65 mmol e- kg-1 range), whereas the CDAC of bergamot SPJ (816 mmol e- kg-1) was strikingly higher than the counterparts from other citrus fruits. This value approached that of bergamot peel extracts (822 mmol e- kg-1). Bergamot peel and SPJ also exhibited the highest DPPH inhibition. The CDAC values were associated with the HPLC-determined content of flavonoids, namely neoeriocitrin, naringin, and neohesperidin, which were 4-10-fold more concentrated in bergamot SPJ and peel than in SPJ from other citrus species. These findings contribute to point at bergamot by-products as rich sources of antioxidant compounds on a quantitative basis, highlighting their enormous potential for pharmaceutical, nutraceutical and food applications.

New Mater-Bi, Biodegradable Mulching Film for Strawberry (Fragaria × Ananassa Duch.): Effects on Film Duration, Crop Yields, Qualitative, and Nutraceutical Traits of Fruits.

In the main strawberry areas of Southern Italy, cultivation is carried out by transplanting plants on raised beds (30-40 cm from ground level), mulched with black polyethylene (PE). This technique has becoming increasingly expensive due to the growing prices of plastic mulches, the cost to remove them at the end of crop cycle, and the difficulty to dispose of black, dirty plastic films. The main objective of this research was the replacement of PE mulch with a new biodegradable mulching film Mater-Bi®-based (Novamont), characterized by an increased permanence in the field designed for long crop life. In 2021, two Mater-Bi-based, black, 18 µm thick mulching films were tested under tunnel: N5 as innovative film and N18 as commercial standard film. Black PE film, 50 µm thick was the control. Strawberry cultivars 'Sabrina' and 'Elide' were cropped on the three mulching films according to a split plot design with four replications. Harvests lasted from March to June 2021. Cvs Sabrina and Elide yielded around 40 t ha-1, while the mean effect of mulching films did not point out differences between the biodegradable mulches and PE. In 4 out of 12 harvests we analyzed samples of fruits to assess the influence of mulches on the contents of °Brix, polyphenols, antioxidant activity, ascorbic acid, flavonoids, and anthocyanins. On average, °Brix was clearly improved in fruits on PE compared to biodegradable films, while all the other qualitative traits resulted in being more dependent on the cultivars and times of sampling effects. Overall, biodegradable mulches are a viable alternative to PE mulch, and the innovative N5 film appeared promising for the enhancement of durability of soil coverage in a long-lasting cycle.

In vitro digestion of milk proteins including intestinal brush border membrane peptidases. Transepithelial transport of resistant casein domains.

The use of enzymes from the brush border membrane (BBM) in simulated gastrointestinal digestion of milk proteins has been evaluated. With this purpose, the resistant sequences from casein and milk whey proteins after INFOGEST in vitro digestion with and without BBM have been analyzed by tandem mass spectrometry. The use of BBM revealed additional cleavages to those found with pancreatic enzymes, although the number of total identified peptides decreased due to the reduction of the peptide size. These new cleavages were mainly attributed to the activity of amino- and carboxy-peptidases, which was also reflected in the higher concentration of free amino acids found in the gastrointestinal digests with BBM. The peptidome of the simulated gastrointestinal digests was compared with that previously obtained in digests aspirated from human jejunum after oral administration of the same substrates. The addition of BBM did not change significantly the peptide profile, although it allowed the identification of peptides found in human digests. However, none of the models was able to reproduce the large variety of peptides found in vivo. In addition, in vitro transepithelial transport of six ß-casein derived peptides resistant to gastrointestinal digestion, including the opioid ß-casomorphin-7, was also evaluated. The results point to the importance of the nature of the N- and C-terminal end for the transport rate through the Caco-2 cell monolayer. Therefore, the use of BBM as a supplementary step after simulated pancreatic digestion can be considered in bioavailability studies since the final sequence can determine the absorption of peptides.

Rapid and Non-Destructive Techniques for the Discrimination of Ripening Stages in Candonga Strawberries.

Electronic nose (e-nose), attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy and image analysis (IA) were used to discriminate the ripening stage (half-red or red) of strawberries (cv Sabrosa, commercially named Candonga), harvested at three different times (H1, H2 and H3). Principal component analysis (PCA) performed on the e-nose, ATR-FTIR and IA data allowed us to clearly discriminate samples based on the ripening stage, as in the score space they clustered in distinct regions of the plot. Moreover, a correlation analysis between the e-nose sensor and 57 volatile organic compounds (VOCs), which were overall detected in all the investigated fruit samples by headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry (HS-SPME/GC-MS), allowed us to distinguish half-red and red strawberries, as the e-nose sensors gave distinct responses to samples with different flavours. Three suitable broad bands were individuated by PCA in the ATR-FTIR spectra to discriminate half-red and red samples: the band centred at 3295 cm-1 is generated by compounds that decline, whereas those at 1717 cm-1 and at 1026 cm-1 stem from compounds that accumulate during ripening. Among the chemical parameters (titratable acidity, total phenols, antioxidant activity and total soluble solid) assayed in this study, only titratable acidity was somehow correlated to ATR-FTIR and IA patterns. Thus, ATR-FTIR spectroscopy and IA might be exploited to rapidly assess titratable acidity, which is an objective indicator of the ripening stage.

Proteomics and Integrated Techniques to Characterize Organic Residues in Funerary Findings from Italic Populations of the First Millennium BC.

Multiple analytical techniques were combined to achieve a detailed characterization of organic residues in different typologies of funerary pottery, which were found at two separate archeological sites in the Campania Region (Italy) and both dated back to the first millennium BC. Gas chromatography-mass spectrometry (GC-MS) analysis of lipids provided inconclusive results. The attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectra of encrustation on two glazed bowls of the 3rd to 4th century BC were comparable to those of fresh bone, revealing the presence of hydroxyapatite and proteins, which were identified as bovine collagen chains by liquid chromatography coupled to high-resolution tandem mass spectrometry (LC-MS/MS)-based proteomics. This finding confirmed that Italic populations used to inhume the dead along with votive meat offerings. Proteomics was decisive for identifying bovine milk in an unusually shaped amphora unearthed from a grave that belonged to a woman at the necropolis of the Greek colony in Cuma (7th century BC). Peptidomic analysis demonstrated that the genetic variant A1 of ß-casein was already present in the southern Mediterranean area at least 2500 years ago. Overall, these results depict an agropastoral system of Italic populations at the age of Magna Graecia based on a significant role of domesticated cattle.

In vivo absorptomics: Identification of bovine milk-derived peptides in human plasma after milk intake.

A dedicated two-step purification procedure prior to nanoliquid chromatography-electrospray-tandem mass spectrometry analysis enabled the identification of bovine milk-derived peptides absorbed and circulating in the plasma of three healthy volunteers who received 250 mL of pasteurized milk after a 10-days washout. The appearance and clearance of milk peptides in plasma were monitored at various time points. Overall, 758, 273 and 212 unique peptides derived from 15, 15 and 18 bovine milk proteins, respectively, were identified in the plasma of these volunteers, evidencing a substantial inter-individual variability. Peptides encrypting possible bioactive and/or immunogenic molecules originating from caseins, ß-lactoglobulin and minor milk proteins were detected. Peptide representation data revealed the combined action of endoproteases involved in primary hydrolysis during gastroduodenal digestion and exopeptidases that hydrolyse peptides in the small intestine. It remains to be established whether the half-life and concentration ranges of circulating milk-derived peptides may have any impacts on human health.

Selection of Lactiplantibacillus Strains for the Production of Fermented Table Olives.

Lactiplantibacillus strains (n. 77) were screened for technological properties (e.g., xylose fermentation, EPS production, antimicrobial activity, tolerance to NaCl and phenolic compounds, oleuropein degradation and hydroxytyrosol formation) relevant for the production of fermented table olives. Survival to olive mill wastewater (OMW) and to simulated gastro-intestinal tract (GIT), the capability to grow at different combinations of NaCl and pH values, radical scavenging activities and biofilm formation were further investigated in 15 selected strains. The screening step revealed high diversity among Lactiplantibacillus strains. Most of the strains were able to ferment xylose, while only a few strains produced EPS and had inhibitory activity against Y. lipolytica. Resistance to phenolic compounds (gallic, protocatechuic, hydroxybenzoic and syringic acids), as well as the ability to release hydroxytyrosol from oleuropein, was strain-specific. OMWs impaired the survival of selected strains, while combinations of NaCl ≤ 6% and pH ≥ 4.0 were well tolerated. DPPH and hydroxyl radical degradation were strain-dependent, while the capability to form biofilm was affected by incubation time. Strains were very tolerant to the GIT. The genome of Lpb. pentosus O17 was sequenced and analysed to verify the presence of genes involved in the degradation and metabolism of phenolic compounds. O17 lacks carboxylesterase and gallate decarboxylase (subunits B and D) sequences, and its gene profile differs from that of other publicly available Lpb. pentosus genomes.

Effect of sprouting on the proteome of chickpea flour and on its digestibility by ex vivo gastro-duodenal digestion complemented with jejunal brush border membrane enzymes.

The demand for sustainably produced proteins is increasing with the world population and is prompting a dietary shift toward plant sourced proteins. Vegetable proteins have lower digestibility and biological value compared to animal derived counterparts. We explored sprouting of chickpea seeds as a strategy for improving digestibility. Protein evolution associated with by the sprouting process was assessed by proteomics. The sprouting induced breakdown of seed storage proteins and doubled the release of free alpha-amino nitrogen in sprouted chickpea flour. During sprouting, several enzymes involved in plant development were newly expressed. An ex vivo model of gastroduodenal and jejunal digestion was applied to assess the bioaccessibility of the protein digests. Proteins from chickpea sprouts showed a greater susceptibility to digestion with a 10% increase in alpha amino nitrogen. Peptides with potential immunoreactivity or bioactivity were catalogued in both digested chickpea sprouts and seeds using an in-silico approach. Peptides belonging to the non-specific transfer proteins, which are allergens in pulses, and peptides belonging to an IgE-binding hemagglutinin protein could only be identified in the digested chickpea sprouts. The observation collected paved the way to immune-based evaluations to assess the effect of germination on the allergenic potential.

Differential Protein Expression in Berry Skin from Red Grapes with Varying Hybrid Character.

Protein expression from the berry skin of four red grape biotypes with varying hybrid character was compared at a proteome-wide level to identify the metabolic pathways underlying divergent patterns of secondary metabolites. A bottom-up shotgun proteomics approach with label-free quantification and MaxQuant-assisted computational analysis was applied. Red grapes were from (i) purebred Vitis vinifera (Aglianico cv.); (ii) V. vinifera (local Sciascinoso cv.) grafted onto an American rootstock; (iii) interspecific hybrid (V. vinifera × V. labrusca, Isabel), and (iv) uncharacterized grape genotype with hybrid lineage, producing relatively abundant anthocyanidin 3,5-O-diglucosides. Proteomics supported the differences between hybrids and purebred V. vinifera grapes, consistently with distinct phenotypic metabolite assets. Methanol O-anthraniloyltransferase, which catalyses the synthesis of methyl anthranilate, primarily responsible for the "foxy" odour, was exclusive of the Isabel hybrid grape. Most of the proteins with different expression profiles converged into coordinated biosynthetic networks of primary metabolism, while many possible enzymes of secondary metabolism pathways, including 5-glucosyltransferases expected for hybrid grapes, remained unassigned due to incomplete protein annotation for the Vitis genus. Minor differences of protein expression distinguished V. vinifera scion grafted onto American rootstocks from purebred V. vinifera skin grapes, supporting a slight influence of the rootstock on the grape metabolism.

Tritordeum as an Innovative Alternative to Wheat: A Comparative Digestion Study on Bread.

Tritordeum results from the crossbreeding of a wild barley (Hordeum chilense) species with durum wheat (Triticum turgidum spp. turgidum). This hexaploid crop exhibits agronomic and rheological characteristics like soft wheat, resulting in an innovative raw material to produce baked goods. We applied a gel-based proteomic approach on refined flours to evaluate protein expression differences among two widespread tritordeum cultivars (Aucan and Bulel) taking as the reference semolina and flour derived from a durum and a soft wheat cvs, respectively. The products of in vitro digestion of model breads were analyzed to compare bio-accessibility of nutrients and mapping tritordeum bread resistant peptides. Significant differences among the protein profiles of the four flours were highlighted by electrophoresis. The amino acid bio-accessibility and the reducing sugars of tritordeum and wheat breads were comparable. Tritordeum cvs had about 15% higher alpha-amino nitrogen released at the end of the duodenal simulated digestion than soft wheat (p < 0.05). Bulel tritordeum flour, bread and digested bread had about 55% less R5-epitopes compared to the soft wheat. Differences in protein expression found between the two tritordeum cvs reflected in diverse digestion products and allergenic and celiacogenic potential of the duodenal peptides. Proteomic studies of a larger number of tritordeum cvs may be successful in selecting those with good agronomical performances and nutritional advantages.

Accurate determination of total biophenols in unfractionated extra-virgin olive oil with the fast blue BB assay.

The phenolic compounds of extra-virgin olive oil (EVOO) are key contributors of nutritional and sensory quality as well as chemical stability. The reference method for their determination is the HPLC-UV, which is cost-/time-expensive. In this work, total phenolic compounds were evaluated in unfractionated EVOO adapting the Fast Blue BB (FBBB) assay, which involves the spectrophotometric (absorbance at 420 nm) determination of azo derivatives resulting from the coupling of phenolic compounds with FBBB diazonium salt in alkali pH. When tested on 26 EVOO samples, the FBBB assay and HPLC-determinations were strikingly correlated (R2 = 0.9653), differently from FBBB and Folin-Ciocalteu assays, which showed poor correlation. The assay is simple, repeatable, robust, rapid and cheap, and results might be evaluated on a printed colorimetric scale. This protocol of the FBBB assay could be routinely used to categorize EVOO according to the health claim allowed by EFSA concerning the content of phenolic compounds.