Protective Efficacy of the Epitope-Conjugated Antigen N-Tc52/TSkb20 in Mitigating Trypanosoma cruzi Infection through CD8+ T-Cells and IFNγ Responses.

Chagas disease, caused by the protozoan Trypanosoma cruzi, remains a major public health challenge affecting millions in Latin America and worldwide. Although significant progress has been made in vector control, no vaccine exists to prevent infection or mitigate disease pathogenesis. We developed a rationally designed chimeric protein vaccine, N-Tc52/TSkb20, incorporating immunodominant epitopes from two T. cruzi antigens, the amino-terminal portion of Tc52 and the TSkb20 epitope derived from trans-sialidase. The objectives of this study were to construct and characterize the antigen and evaluate its protective potential in an immunoprophylactic murine model of T. cruzi infection. The N-Tc52/TSkb20 protein was recombinantly expressed in E. coli and its identity was confirmed using mass spectrometry and Western blotting. Immunization with the chimeric protein significantly controlled parasitemia and reduced the heart, colon, and skeletal muscle parasite burdens compared to non-vaccinated mice. Protection was superior to vaccination with the individual parental antigen components. Mechanistically, the vaccine induced potent CD8+ T-cell and IFNγ responses against the incorporated epitopes and a protective IgG antibody profile. A relatively low IL-10 response favored early parasite control. These results validate the promising multi-epitope approach and support the continued development of this type of rational vaccine design strategy against Chagas disease.

Postharvest Quality of Citrus medica L. (cv Liscia-Diamante) Fruit Stored at Different Temperatures: Volatile Profile and Antimicrobial Activity of Essential Oils.

Citron (Citrus medica L. cv. Liscia-diamante), cultivated in the "Riviera dei Cedri" (southern Italy), is mainly utilized in the production of candied fruit and essential oils (EOs). Up to now, no information regarding the effect of storage temperatures on citron has been reported. Here, citron samples, after harvesting, were stored at different temperatures (5, 10 and 20 °C at 70% relative humidity) for two weeks, and the main postharvest quality parameters were evaluated. Moreover, EOs extracted from the stored samples were chemically characterized to reveal changes in the volatiles profile and antimicrobial activity. The EOs presented monoterpene hydrocarbons (87.1 to 96.3% of the total oil profile) as the most abundant compounds, followed by oxygenated metabolites ranging from 9.7 to 3.1% of the total pattern. Postharvest quality traits showed a good retention of green peel color during storage at 5 °C, while EOs from samples stored for 7 and 14 days at 10 and 20 °C, respectively, showed the highest antimicrobial activity against most assayed strains. The results indicated storage at 10 °C for 7 days as the most suitable for the preservation of the postharvest quality of the fruit and the antimicrobial activity of the extracted EOs.

Sustainable Valorization of Industrial Cherry Pomace: A Novel Cascade Approach Using Pulsed Electric Fields and Ultrasound Assisted-Extraction.

In this study, a two-stage cascade extraction process utilizing pulsed electric fields (PEF) (3 kV/cm, 10 kJ/kg) for initial extraction, followed by ultrasound (US) (200 W, 20 min)-assisted extraction (UAE) in a 50% (v/v) ethanol-water mixture (T = 50 °C, t = 60 min), was designed for the efficient release of valuable intracellular compounds from industrial cherry pomace. The extracted compounds were evaluated for total phenolic content (TPC), flavonoid content (FC), total anthocyanin content (TAC), and antioxidant activity (FRAP), and were compared with conventional solid-liquid extraction (SLE). Results showed that the highest release of bioactive compounds occurred in the first stage, which was attributed to the impact of PEF pre-treatment, resulting in significant increases in TPC (79%), FC (79%), TAC (83%), and FRAP values (80%) of the total content observed in the post-cascade PEF-UAE process. The integration of UAE into the cascade process further augmented the extraction efficiency, yielding 21%, 49%, 56%, and 26% increases for TPC, FC, TAC, and FRAP, respectively, as compared to extracts obtained through a second-stage conventional SLE. HPLC analysis identified neochlorogenic acid, 4-p-coumaroylquinic, and cyanidin-3-O-rutinoside as the predominant phenolic compounds in both untreated and cascade-treated cherry pomace extracts, and no degradation of the specific compounds occurred upon PEF and US application. SEM analysis revealed microstructural changes in cherry pomace induced by PEF and UAE treatments, enhancing the porosity and facilitating the extraction process. The study suggests the efficiency of the proposed cascade PEF-UAE extraction approach for phenolic compounds from industrial cherry pomace with potential applications to other plant-based biomasses.

Volatile Metabolites to Assess the Onset of Chilling Injury in Fresh-Cut Nectarines.

Fresh-cut processing is a good strategy to enhance the commercialization of peaches and nectarines, which easily deteriorate during low-temperature storage mostly due to the occurrence of chilling injury. Although several studies have been performed to improve the shelf-life of fresh-cut stone fruit, the achievement of high-quality fresh-cut peaches and nectarines still constitutes a challenge. The present study aimed to gain insights into the evolution of the postharvest quality of fresh-cut nectarines (Prunus persica L. Batsch) Big Bang, cold-stored at two different storage temperatures (4 and 8 °C) for up to 10 days. Several aspects influencing the quality traits (sensory and postharvest quality parameters; the profile of phenolic and volatile organic compounds (VOCs)) were explored to predict the marketable life of the fresh-cut nectarines. The respiration rate was higher in samples stored at 4 °C, while the browning process was more evident in fruit stored at 8 °C. Partial Least Squares Regression performed on VOCs showed that samples stored at 4 °C and 8 °C presented a different time evolution during the experiment and the trajectories depended on the interaction between time and temperature. Moreover, Multiple Linear Regression analysis discovered that the 17 VOCs affected by the storage conditions seemed to suggest that no chilling injury was detected for nectarines Big Bang. In conclusion, this approach could also be used with other nectarine cultivars and/or different stone fruits.

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.

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.

Immunometabolism Modulation by Extracts from Pistachio Stalks Formulated in Phospholipid Vesicles.

Several studies have demonstrated the effectiveness of plant extracts against various diseases, especially skin disorders; namely, they exhibit overall protective effects. The Pistachio (Pistacia vera L.) is known for having bioactive compounds that can effectively contribute to a person's healthy status. However, these benefits may be limited by the toxicity and low bioavailability often inherent in bioactive compounds. To overcome these problems, delivery systems, such as phospholipid vesicles, can be employed. In this study, an essential oil and a hydrolate were produced from P. vera stalks, which are usually discarded as waste. The extracts were characterized by liquid and gas chromatography coupled with mass spectrometry and formulated in phospholipid vesicles intended for skin application. Liposomes and transfersomes showed small size (<100 nm), negative charge (approximately -15 mV), and a longer storage stability for the latter. The entrapment efficiency was determined via the quantification of the major compounds identified in the extracts and was >80%. The immune-modulating activity of the extracts was assayed in macrophage cell cultures. Most interestingly, the formulation in transfersomes abolished the cytotoxicity of the essential oil while increasing its ability to inhibit inflammatory mediators via the immunometabolic citrate pathway.

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.

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.

Fluorescence Detecting of Paraquat and Diquat Using Host-Guest Chemistry with a Fluorophore-Pendant Calix[6]arene.

Paraquat (PQ) and diquat (DQ), some of the most widely used herbicides in the world, both present a high mortality index after intentional exposure. In this paper, a fluorescence sensing method for PQ and DQ, based on host-guest molecular recognition, is proposed. Calix[6]arene derivatives containing anthracene or naphthalene as pendant fluorophore at their lower rim recognize DQ and PQ in hydroalcoholic solution with a broad linear response range at the µg L-1 level concentration. The linear response ranges were found from 1.0 to 18 µg L-1 with the detection limit of 31 ng L-1 for paraquat, and from 1.0 to 44 µg L-1 with the detection limit of 0.16 µg L-1 for diquat. The recognition process is detected by following the decrease in the fluorescence emission consequent to complexation. The proposed quenching method has been applied to the determination of paraquat in drinking water samples.

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.

E40 glutenase detoxification capabilities of residual gluten immunogenic peptides in in vitro gastrointestinal digesta of food matrices made of soft and durum wheat.

Gluten degrading enzymes, which are commonly referred to as "glutenases," represent attractive candidates for the development of a pharmacological treatment of gluten related disorders, such as coeliac disease (CeD). Endoprotease-40 (E40), a novel glutenase secreted by the actinomycete Actinoallomurus A8 and recombinantly produced in S. lividans TK24, was shown to be active at pH 3 to 6 (optimum pH 5), resistant to pepsin and trypsin degradation, able to destroy immunotoxicity of both gliadin 33-mer peptide and whole proteins and to strongly reduce the response of specific T cells when added to gliadin in in vitro gastrointestinal digestion. This study aims to functionally assess the capabilities of Endoprotease-40 (E40) to detoxify residual gluten immunogenic peptides in gastrointestinal digesta of food matrices made of soft and durum wheat. The INFOGEST harmonized protocols were applied to the multicompartmental model of simulated human gastrointestinal digestion, for the quantitative assessment of residual gluten in liquid (beer) and solid (bread and pasta) foods, made of either soft or durum wheat. Proteomic and immunological techniques, and functional assays on intestinal T cell lines from celiac disease patients were used to identify gluten-derived immunogenic peptide sequences surviving in gastric and gastrointestinal digesta after the addition of E40 at increasing enzyme: wheat proteins ratios. During the gastric phase (2 h incubation time), the addition of E40 demonstrated an extensive (≥ 95%) dose-dependent detoxification of whole gluten in real food matrices. Overall, the residual gluten content was found at, or even below, the 20 ppm gluten-free threshold for soft and durum wheat-based food. Furthermore, unlike in untreated gastrointestinal digesta, none of the immunodominant α-gliadin peptides survived in E40-treated digesta. Traces of ω- and γ-gliadin derived immunogenic peptides were still detected in E40-treated digesta, but unable to stimulate celiac-intestinal T cells. In conclusion, E40 is a promising candidate for the oral enzymatic therapy of CeD, as a stand-alone enzyme being efficient along the complete gastrointestinal digestion of gluten.

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.

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.

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.

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.

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.

Profiles of Volatile and Phenolic Compounds as Markers of Ripening Stage in Candonga Strawberries.

Volatile compounds, quality traits (total phenols and antioxidant capacity) and High-performance liquid chromatography (HPLC)-isolated polyphenols of strawberries, variety Sabrosa, commercially referred to as "Candonga", harvested at three different times (H1, H2 and H3) and at two different ripening stages, namely half-red (Half-red-H1, Half-red-H2 and Half-red-H3) and red (Red-H1, Red-H2 and Red-H3) were evaluated. Dominant anthocyanins, namely cyanidin-3-O-glucoside, pelargonidin-3-O-glucoside and pelargonidin-3-O-rutinoside, as well as p-coumaryl hexoside increased during harvesting, differently from flavonoids, such as quercetin-3-O-glucoside, kaempferol-3-O-glucoronide and quercetin 3-O-glucoronide, that declined. Samples clustered in different quadrants of the principal component analysis (PCA) performed on volatiles, quality traits and phenolic compounds, highlighting that only the red samples were directly correlated to volatile components, as volatiles clearly increased both in number and amount during ripening. In particular, volatiles with a positive impact on the consumers' acceptance, including butyl butyrate, ethyl hexanoate, hexyl acetate, nonanal, terpenes and lactones, were positively associated with the Red-H1 and Red-H2 strawberries, while volatiles with negative coefficients related to consumer liking, including isopropyl butyrate, isoamyl butyrate and mesifurane directly correlated with the Red-H3 samples. Accordingly, strawberries harvested at Red-H1 and Red-H2 ripening stages could be preferred by the consumers compared to the Red-H3 fruit. Altogether, these results could help to individuate quality traits as putative markers of the ripening stage, and optimize the process of post-harvesting ripening to preserve or improve the desirable aromatic characteristics of strawberries.