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Coppa Piacentina is an Italian protected designation of origin (PDO) dry-cured product obtained from the muscle of pork neck and ripened for at least six months. Metabolomics- and volatilomics-based strategies, combined with a chemical characterization of free amino acids were applied to identify biomarkers of long-ripened Coppa Piacentina PDO. Long ripening induced a significantly increase of total free amino acids, mainly represented by glutamic acid, involved in the umami taste perception. Untargeted metabolomics, performed using UHPLC-HRMS, allowed to identify 32 putative gamma-glutamyl-peptides, known as main contributors to the kokumi taste. Unsupervised and supervised multivariate statistics observed a clear modification of these peptides over the ripening, with gamma-glutamyl-peptides which significantly increased in long-ripened samples. A volatilomics-based strategy, conducted with GCxGC-MS, was then performed, and 93 different compounds were identified, with aldehyde and ketones deriving from the lipid auto-oxidation which increased according to ripening.

An untargeted metabolomics approach combined with sensory analysis was used to depict the impact of different traditional Italian extraction methods (i.e., Espresso, Neapolitan, Moka) along with Filter, on Coffea arabica and Coffea canephora var. robusta beverages. To this aim, polyphenols, Maillard reaction products, and coffee metabolites were screened by high resolution mass spectrometry and elaborated through both unsupervised and supervised multivariate statistical approaches. Multivariate statistics showed a distinctive chemical profile for Espresso preparation, while Moka and Neapolitan were very similar. The orthogonal projection to latent structures and discriminant analysis allowed the identification of 86 compounds showing a high VIP discrimination score (i.e., > 0.8). The 2,5-dimethyl-3-(methyldithio)-furan was a marker for the Filter preparation, while 1,2-disinapoylgentiobiose characterized both Filter and Neapolitan extractions. Caffeine (known to be a bitter compound) accumulated highly in Filter vs. Espresso, although at the sensory profile, bitterness was more perceived in Espresso. Vegetal aroma carried by pyrazines, pyridines, and phenolic acids were markers of Espresso, with Robusta showing higher values than Arabica. Notwithstanding, our findings showed that the extraction process played a hierarchically higher role in driving the chemical composition of the beverages when compared to coffee species.

Positive effects have been observed as a result of Aloe arborescens supplementation in the dry-off phase in dairy cows. Metabolomic approaches can provide additional information about animal physiology. Thus, we characterized plasma metabolome around dry-off in 12 cows supplemented (AL) or not (CTR) with 10 g/d of lyophilized A. arborescens with an untargeted metabolomic approach. Overall, 1658 mass features were annotated. Regardless of treatment, multivariate statistics discriminated samples taken before and after dry-off. Overall, 490 metabolites were different between late lactation and early dry period, of which 237 were shared between AL and CTR. The most discriminant compounds (pentosidine and luteolin 7-O-glucoside) were related to the more fibrous diet. Pathway analysis indicated that pyrimidine and glycerophospholipid metabolisms were down-accumulated, suggesting reduced rumen microbial activity and liver load. Samples from AL were discriminated from CTR either the day of dry-off or 7 days after. At dry-off, aloin and emodin were the most discriminant metabolites, indicating that Aloe's bioactive compounds were absorbed. Seven days later, 534 compounds were different between groups, and emodin was among the most impacted. Pathway analysis highlighted that glycerophospholipid, pyrimidine, and folate metabolisms were affected. These results might indicate that Aloe has positive effects on liver function and a modulatory effect on rumen fermentation.

Thymus comosus Heuff ex. Griseb. (Lamiaceae) is a wild thyme species endemic for Romanian Carpathian areas, frequently collected as substitute for collective herbal product Serpylli herba, cited as antibacterial and diuretic remedy in traditional medicine. The present study aimed to evaluate the in vivo diuretic effect and in vitro antimicrobial properties of three herbal preparations (infusion—TCI, tincture—TCT and an hydroethanolic extract prepared through an optimized ultrasound-assisted method—OpTC) obtained from the aerial parts of T. comosus Heuff ex. Griseb, also evaluating their comprehensive phenolic profile. In vivo diuretic effect was tested using Wistar rats treated orally with each herbal preparation (125 and 250 mg/kg dispersed in 25 ml/kg isotonic saline solution) and quantified based on cumulative urine output (ml), diuretic action and diuretic activity. Additionally, sodium and potassium excretion were monitored using a potentiometric method with selective electrodes. In vitro antibacterial and antifungal activities were assessed using p-iodonitrotetrazolium chloride assay against six bacterial strains and six fungal strains by monitoring minimum inhibitory concentration (MICs), minimum bactericidal concentrations (MBCs) and minimum fungicidal concentrations (MFCs). Finally, phenolic profile of the aforementioned herbal extracts was evaluated using an ultra-high-pressure liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) method to check the impact of the different preparations on the most abundant and significant compounds. All the extracts exerted a mild diuretic action, TCT and OpTC inducing the most intense diuretic effect. Both herbal preparations produced a statistically significant, dose-dependent and gradual increase of the urine output, the effect being more intense at 24 h (6.63–7.13 ml/24 h). Potentiometric evaluation of urine samples collected from treated rats revealed a clear and mild

This work aimed to investigate the influence of high-power ultrasound (HPU) technology on the stability of bioactive compounds in strawberry juices obtained from fruits with different stages of ripeness (75% vs. 100%) and stored at 4 degrees C for 7 days. HPU parameters were amplitude (25, 50, 75, and 100%), pulses (50 vs. 100%) and treatment time (5 vs. 10 min). Amplitude and pulse had a significant effect (p <= 0.05) on all bioactive compounds except flavonols and hydroxycinnamic acids. The treatment duration of 5 min vs. 10 min had a significant positive impact on the content of anthocyanins, flavonols and condensed tannins, while the opposite was observed for total phenols, whereas no statistically significant effect was observed for hydroxycinnamic acids. The temperature changes during HPU treatment correlated positively with almost all HPU treatment parameters (amplitude, pulse, energy, power, frequency). Optimal parameters of HPU were obtained for temperature changes, where the highest content of a particular group of bioactive compounds was obtained. Results showed that by combining fruits with a certain ripeness and optimal HPU treatment, it would be possible to produce juices with highly preserved bioactive compounds, while HPU technology has prospects for application in functional food products.

Untargeted metabolomics based on ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry was combined with sensory analysis to provide new insights into the impact of the feeding system from mountain regions (grassland deriving from permanent meadows) on the chemical fingerprint of Parmigiano Reggiano PDO hard cheese. In the framework of a representative investigation, two different ripening times (12 and 24 months) were also considered. Multivariate statistics allowed discriminating cheese samples from different feeding regimens according to their metabolomics signatures. Interestingly, mountain grassland-based cheese samples were characterized by a more favourable fatty acid profile, recording also feed-derived compounds (such as terpenoids and linoleic acid derivatives) potentially associated with both beneficial effects on human health and sensory properties. According to the sensory analysis, the impact of herbs and grass enhanced the colour and retro-olfactive complexity of Parmigiano Reggiano PDO cheese, with spicy, umami and intense vegetal aromatic notes representing distinctive features.

Dairy propionibacteria are Gram positive Actinomycetota, routinely utilized as starters in Swiss type cheese making and highly appreciated for their probiotic properties and health promoting effects. In this work, within the frame of a circular economy approach, 47 Propionibacterium and Acidipropionibacterium spp. were isolated from goat cheese and milk, and ewe rumen liquor, and characterized in view of their possible utilization for the production of novel pro-bioactive food and feed on scotta, a lactose rich substrate and one of the main by-products of the dairy industry. The evaluation of the Minimum Inhibitory Concentration (MIC) of 13 among the most common antibiotics in clinical practice revealed a general susceptibility to ampicillin, gentamycin, streptomycin, vancomycin, chloramphenicol, and clindamycin while confirming a lower susceptibility to aminoglycosides and ciprofloxacin. Twenty-five isolates, that proved capable of lactose utilization as the sole carbon source, were then characterized for functional and biotechnological properties. Four of them, ascribed to Propionibacterium freudenreichii species, and harboring resistance to bile salts (growth at 0.7-1.56 mM of unconjugated bile salts), acid stress (>80% survival after 1 h at pH 2), osmostress (growth at up to 6.5% NaCl) and lyophilization (survival rate > 80%), were selected and inoculated in scotta. On this substrate the four isolates reached cell densities ranging from 8.11 & PLUSMN; 0.14 to 9.45 & PLUSMN; 0.06 Log CFU mL(-1) and proved capable of producing different vitamin B9 vitamers after 72 h incubation at 30 & DEG;C. In addition, the semi-quantitative analysis following the metabolomics profiling revealed a total production of cobalamin derivatives (vitamin B12) in the range 0.49-1.31 mg L-1, thus suggesting a full activity of the corresponding biosynthetic pathways, likely involving a complex interplay between folate cycle and methylation cycle required in vitamin B12 b

In the last years, animal proteins from meat and meat-derived products have represented one of the main concerns to achieve a sustainable food production. Within this perspective, the partial replacement of meat with nonmeat substances having a high-protein content, offers interesting opportunities to reformulate more sustainable meat products characterized also by potential health-promoting properties. Therefore, starting from these background conditions, this short review critically summarizes recent findings on extenders from different origins, namely pulses, plant-based ingredients, plant by-products, and unconventional sources, as valuable possibility to improve the technological profile and functional quality of meat, together with providing a focus on their ability to affect the sustainability of meat products.

IDPlanT is the Italian Database of Plant Translocation, an initiative of the Nature Conservation Working Group of the Italian Botanical Society. IDPlanT currently includes 185 plant translocations. The establishment of a national database on plant translocation is a key step forward in data sharing and techniques improvement in this field of plant conservation. Supplemental data for this article is available online at https://doi.org/10.1080/11263504.2021.1985004.

Parmigiano Reggiano is a hard cheese with a Protected Designation of Origin (PDO) certification that also applies to the grated product. The percentage of rind in grated Parmigiano Reggiano is regulated by the PDO production Specification and must not exceed the limit of 18% (w/w). The present study evaluates the potential of an untargeted foodomics approach to detect anomalous inclusions of rind in grated Parmigiano Reggiano cheese. In particular, a combined metabolomics and peptidomics approach was used to detect potential markers of counterfeits (rind > 18%). In the framework of realistic food integrity purposes, non-Parmigiano Reggiano grated samples and different ripening times were also considered. Untargeted metabolomics allowed detecting 347 compounds, with a prevalence of amino acids and peptide derivatives, followed by fatty acyls and other compounds (such as lactones, ketones, and aldehydes) typically related to proteolysis and lipolysis events. Overall, the unsupervised multivariate statistics showed that the ripening time plays a hierarchically higher impact than rind inclusion in determining the main differences in the chemical profiles detected. Interestingly, supervised statistics highlighted distinctive markers for ripening time and rind inclusion, with only 16 common discriminant compounds being shared between the two conditions. The best markers of rind inclusion > 18% were 2-hydroxyadenine (VIP score = 1.937; AUC value = 0.83) and the amino acid derivatives argininic acid (VIP score = 1.462; AUC value = 0.75) and 5-hydroxyindole acetaldehyde (VIP score = 1.710; AUC value = 0.86). Interestingly, the medium-chain aldehyde 4-hydroperoxy-2-nonenal was a common marker of both ripening time and anomalous rind inclusion (>18%), likely arising from the lipid oxidation processes. Finally, among potential marker peptides of rind inclusion, the alpha-S1 casein proteolytic product (F)FVAPFPEVFGK(E) could be identified.

The consumption of foods characterized by a wide bioactive-compound profile has been associated with improved health status and reduced risk of developing diseases. Owing to the complexity of food production (e.g. crops and farmed animals), processing (e.g. fermentation), and shelf life (e.g. lipid or protein oxidation), a more comprehensive and holistic analysis is necessary to investigate the intricate metabolic pathways and metabolites involved with or displaying a biological activity. Accordingly, untargeted metabolomics analysis can provide crucial knowledge to support the selection of key metabolites and metabolic pathways to produce foods from animals and crops with a functional value. This short review aims to provide an overview of the applications of metabolomic analysis to screen bioactive compounds in both animal- and plant-based foods.

Saffron production from Crocus sativus flowers produces large amounts of by-products that may represent an excellent source of polyphenols. The aim of this work was to evaluate infusions originating from different brewing processes and from different saffron flower portions, in terms of both functional and sensory traits. For this aim, total polyphenols and total flavonoids, in vitro antioxidant assays and an untargeted phenolic profiling were applied. In general, tepals showed higher polyphenol and flavonoid content than stamen infusions, and their bioactive content depended more on brewing temperature than brewing time. These findings were consistent with both antioxidant capacity and phenolic profiling. Multivariate statistics highlighted polyphenols discriminating ‘boiled’ vs. ‘cold’ infusions, being mainly flavonoids, phenolic acids and the alkylphenol 5-pentadecylresorcinol (showing a strong down-accumulation at the higher brewing temperatures). Positive correlations could be highlighted between anthocyanins, flavones, flavonols and lignans, and the in vitro antioxidant assays. In general, cold brewing was successful in extracting phenolic compounds and provided better sensory properties, thus indicating that this may represent a valuable strategy to develop saffron-based functional beverages with better consumers' acceptability.

Several studies have demonstrated that stigmas of Crocus sativus contain several bioactive compounds with potential health-promoting properties. However, during the processing of stigmas, large amounts of floral bio-residues are normally discarded as by-products. In this study, using untargeted metabolomics, the comprehensive phytochemical composition of C. sativus stigma and tepals was investigated. Moreover, the antibacterial and anti-biofilm properties of the extracts of C. sativus stigmas and tepals were compared. The study was carried out using two methicillin-resistant staphylococcal reference strains (i.e., Staphylococcus aureus ATCC 43300 and Staphylococcus epidermidis ATCC 35984), representing important Gram-positive biofilm-forming human pathogens. The antibacterial properties were correlated with total polyphenol content, total terpenoid content, and in vitro antioxidant properties of tepals and stigmas. The results demonstrated that stigma and tepal extracts, at the sub-Toxic concentrations, were able to interfere with biofilm formation by ATCC 43300 and ATCC 35984. Besides, the higher antibacterial activity of tepals than stigmas was associated with higher levels of phycompounds. Therefore, our results demonstrated that C. sativus stigmas and bio-residues, such as tepals, are potential antioxidant sources and good candidates as antibacterial agents to prevent biofilm formation. Taken together, these findings showed that C. sativus could be used as functional ingredient by the food and pharmaceutical industries.

An optimized ultrasound-assisted extractive method was developed to obtain a polyphenol-enriched extract from the aerial parts of Thymus comosus Heuff. ex Griseb. et Schenk. Optimization process was conducted based on Design of Experiment (DoE) principles, determining the influence of three independent variables (time, ultrasound amplitude, ethanol concentration) on the total phenolic content of the extract (dependent variable). Additionally, the phenolic composition of the extract was characterized through UHPLC-HRMS, revealing beside the most abundant flavonoid-type compounds the presence of salvianolic acids C, D and L in high amounts. Phytochemical profile of the extract was correlated with its antioxidant activity (tested through five complementary assays) and enzyme-inhibitory potential, showing important antiglucosidase and anticholinesterase effects. Overall, it was concluded that the developed method is suitable for obtaining a good recovery of both phenolic and non-phenolic compounds from Thymus comosus aerial parts, and their presence in the optimized extract is responsible for its pharmacological potential.

Spices, widely used to improve the sensory characteristics of food, contain several bioactive compounds as well, including polyphenols, carotenoids, and glucosynolates. Acting through multiple pathways, these bioactive molecules affect a wide variety of cellular processes involved in molecular mechanisms important in the onset and progress of human diseases. Capparis spinosa L. is an aromatic plant characteristic of the Mediterranean diet. Previous studies have reported that different parts (aerial parts, roots, and seeds) of C. spinosa exert various pharmacological activities. Flower buds of C. spinosa contain several bioactive compounds, including polyphenols and glucosinolates. Two different subspecies of C. spinosa L., namely, C. spinosa L. subsp. spinosa, and C. spinosa L. subsp. rupestris, have been reported. Few studies have been carried out in C. spinosa L. subsp. rupestris. The aim of our study was to investigate the phytochemical profile of floral buds of the less investigated species C. spinosa subsp. rupestris. Moreover, we investigated the effect of the extract from buds of C. spinosa subsp. rupestris (CSE) on cell proliferation, intracellular ROS levels, and expression of the antioxidant and anti-apoptotic enzyme paraoxonase-2 (PON2) in normal and cancer cells. T24 cells and Caco-2 cells were selected as models of advanced-stage human bladder cancer and human colorectal adenocarcinoma, respectively. The immortalized human urothelial cell line (UROtsa) and human dermal fibroblast (HuDe) were chosen as normal cell models. Through an untargeted metabolomic approach based on ultra-high-performance liquid chromatography quadrupole-time-of-flight mass spectrometry (UHPLC-QTOF-MS), our results demonstrate that C. spinosa subsp. rupestris flower buds contain polyphenols and glucosinolates able to exert a higher cytotoxic effect and higher intracellular reactive oxygen species (ROS) production in cancer cells compared to normal cells. Moreover, upregulation of

In this work, milk samples collected in a cohort of intensive dairy farms of the Po Valley (Italy) were screened for their (poly)-phenolic profile to check the occurrence of phenolic metabolites of biological interest. The selected dairy farms were previously classified on the basis of their cow feeding system, considering the utilization of corn silage as the main ingredient of the rations. Overall, ultra-high-pressure liquid chromatography coupled with mass spectrometry using an Orbitrap analyzer, followed by unsupervised and supervised statistics, allowed identifying clear different phenolic distributions in the milk samples. Accordingly, a great variability in the phenolic profiles of the different milk samples was observed, with two main phenolic clusters outlined by the unsupervised hierarchical clustering approach and not fully correlated to the nutritional strategy considered. The variables’ importance in the projection approach allowed selecting the most important metabolites, resulting in samples’ discrimination. Among the most discriminative compounds, we found phenolic metabolites (such as hippuric acid and 4-hydroxyhippuric acid), followed by lignans (such as enterolactone) and isoflavonoids (such as equol and O-desmethylangolensin). Taken together, our findings suggested that both the feeding systems and the ability of dairy cows to process parent phenolic compounds were the main factors providing the final (poly)-phenolic profile of the milk samples. Future targeted and ad hoc studies appear of great interest to evaluate the potential biological effects of these compounds on cow health

The untargeted metabolomic profile of five pigmented whole rice varieties, namely Ermes, Orange, Nerone, Venere, and Violet, was investigated. Samples were cooked to a comparable starch gelatinization level through an electric commercial rice cooker and in vitro digested to evaluate the fate of rice compounds during in vitro starch digestion. The untargeted metabolomics profiling depicted a broad phytochemical profile of rice grains, including phenolic compounds, terpenoids, and phytosterols, among others. Interestingly, the metabolomic profile showed distinctive shaping following cooking and in vitro digestion in a variety-dependent manner. Multivariate statistics revealed that glycosylated flavonoids and hydrolysis-products of tannins (such as 1,2-digalloyl-beta-D-glucopyranose) were the most discriminant compounds of the in vitro gastrointestinal digestion. The combination of cooking and simulated digestion was found to affect, in a matrix-driven manner, the profile of amino acids and lipids derivatives, together with sterols and terpenoids. Cooked Violet rice grains showed the lowest starch hydrolysis index (i.e., 58.2) and the highest resistant starch content (i.e., 4.8 g/100 g) compared to the other samples. Non-pigmented phenolic compounds could be included among discriminant compounds. After combining cooking and enzyme hydrolysis, Violet rice distinctively showed positive average LogFC values for flavonoids and phenolic acids. Overall, water-soluble anthocyanins were the most retained class of polyphenols following the cooking process, with peonidin 3-(6”-acetyl-glucoside) showing the highest discriminant potential (VIP score = 1.64).

Resurrecting extinct species is a fascinating and challenging idea for scientists and the general public. Whereas some theoretical progress has been made for animals, the resurrection of extinct plants (de-extinction sensu lato) is a relatively recently discussed topic. In this context, the term ‘de-extinction’ is used sensu lato to refer to the resurrection of ‘extinct in the wild’ species from seeds or tissues preserved in herbaria, as we acknowledge the current impossibility of knowing a priori whether a herbarium seed is alive and can germinate. In plants, this could be achieved by germinating or in vitro tissue-culturing old diaspores such as seeds or spores available in herbarium specimens. This paper reports the first list of plant de-extinction candidates based on the actual availability of seeds in herbarium specimens of globally extinct plants. We reviewed globally extinct seed plants using online resources and additional literature on national red lists, resulting in a list of 361 extinct taxa. We then proposed a method of prioritizing candidates for seed-plant de-extinction from diaspores found in herbarium specimens and complemented this with a phylogenetic approach to identify species that may maximize evolutionarily distinct features. Finally, combining data on seed storage behaviour and longevity, as well as specimen age in the novel ‘best de-extinction candidate’ score (DEXSCO), we identified 556 herbarium specimens belonging to 161 extinct species with available seeds. We expect that this list of de-extinction candidates and the novel approach to rank them will boost research efforts towards the first-ever plant de-extinction.

Co-inoculation of arbuscular mycorrhizal fungi (AMF) and bacteria can synergically and potentially increase nitrogen use efficiency (NUE) in plants, thus, reducing nitrogen (N) fertilizers use and their environmental impact. However, limited research is available on AMF-bacteria interaction, and the definition of synergisms or antagonistic effects is unexplored. In this study, we adopted a response surface methodology (RSM) to assess the optimal combination of AMF (Rhizoglomus irregulare and Funneliformis mosseae) and Bacillus megaterium (a PGPR—plant growth promoting rhizobacteria) formulations to maximize agronomical and chemical parameters linked to N utilization in maize (Zea mays L.). The fitted mathematical models, and also 3D response surface and contour plots, allowed us to determine the optimal AMF and bacterial doses, which are approximately accorded to 2.1 kg ha–1 of both formulations. These levels provided the maximum values of SPAD, aspartate, and glutamate. On the contrary, agronomic parameters were not affected, except for the nitrogen harvest index (NHI), which was slightly affected (p-value of < 0.10) and indicated a higher N accumulation in grain following inoculation with 4.1 and 0.1 kg ha–1 of AMF and B. megaterium, respectively. Nonetheless, the identification of the saddle points for asparagine and the tendency to differently allocate N when AMF or PGPR were used alone, pointed out the complexity of microorganism interaction and suggests the need for further investigations aimed at unraveling the mechanisms underlying this symbiosis.

Nutrigenomics is a multidisciplinary science focusing on the diet-genes interaction and exploiting different ‘omics’ technologies to comprehensively investigate the overall effects of different nutrients or diets on health. Recent scientific evidence highlighted the role of dietary polyphenols in modulating gene expression, transcription factors, and cellular processes, thus in potentially affecting health and diseases. Nutrigenomics contributes to the understanding of the molecular mechanisms underlying the direct beneficial effects of polyphenolic compounds in humans. This short review critically summarizes some recent findings on the nutrigenomics-related effects mediated by the major polyphenolic classes of nutritional interest.

Wheat-based vegan muffins were formulated with increasing levels (0, 5 and 10 g per 100 g substitutes to flour) of dried grape pomace powder obtained after distillation (DGPP). The DGPP-muffins showed a greater (P < 0.05) total dietary fibre and a lower (P < 0.05) total starch content to the control. The inclusion of DGPP influenced (P < 0.05) both the batter's rheological properties and the baked muffins' technological characteristics, including the baking loss, the volume, the firmness, the spread ratio and the colour. The in vitro antioxidant activity and the phenolic content increased (P < 0.05) following DGPP inclusion. The untargeted phenolic profiling by UHPLC-HRMS revealed an (P < 0.05) increase of several phenolic classes (i.e. free and bound fractions) following the inclusion of DGPP in the recipe. Flavonoids, mainly anthocyanins, were the most abundant compounds. A quantitative descriptive sensory analysis detected the DGPP-vegan muffins showed good sensory acceptability. The vegan muffin with 5 g per 100 g of DGPP obtained the highest overall acceptability score by the panellists.

Plant cell cultures from cloudberry (CL), lingonberry (LI), stone berry (ST), arctic bramble (AB), and strawberry (SB) were studied in terms of their polyphenol and carotenoid composition, antioxidant activity, antihemolytic activity and cytotoxicity effects on cancerous cells. High-resolution mass spectrometry data showed that LI, presented the highest antioxidant activity, contained the highest contents of flavones, phenolic acids, lignans, and total carotenoids, while CL, ST and SB presented the opposite behavior. AB and SB presented the lowest FRAP and CUPRAC values, while AB and CL presented the lowest reducing power. SB presented the lowest antioxidant activity measured by single electron transfer assays and the lowest content of lignans, phenolic acids, and flavones. CL and LI decreased the viability of in vitro mammary gland adenocarcinoma while only LI decreased the viability of in vitro lung carcinoma and showed protective effects of human erythrocytes against mechanical hemolysis.

The interest in the modulation of gut microbiota by polyphenols from olives and derived products is increasing. In this work, phenolic leaf extracts (PLE) were in vitro faecal fermented to evaluate the changes in phenolic profiles and the impact on microbiota, using a commercial extra-virgin olive oil (EVOO) as reference. The in vitro fermentation decreased oleuropein content in PLE, determining an increase of hydroxytyrosol and other phenolic metabolites. An increase (p < 0.05) of hydroxytyrosol (LogFC = 6.02; VIP score = 1.05) was also observed in fermented EVOO. Besides, PLE significantly (p < 0.05) changed amino acids (LogFC = 6.1) and fatty acids (LogFC = 5.9) profile of the faeces. Metagenomic sequencing revealed that Coriobacteriaceae at the family level, and Collinsella at the genus level, were the most affected by PLE fermentation. These findings support the modulation of the gut microbiota exerted by phenolics from PLE and EVOO.

Wild edible plants have been used in cooking since ancient times. Recently, their value has improved as a result of the scientific evidence for their nutraceutical properties. Sanguisorba minor Scop. (salad burnet) plants were hydroponically grown and two consecutive cuts took place at 15 (C1) and 30 (C2) days after sowing. An untargeted metabolomics approach was utilized to fingerprint phenolics and other health-related compounds in this species; this approach revealed the different effects of the two cuts on the plant. S. minor showed a different and complex secondary metabolite profile, which was influenced by the cut. In fact, flavonoids increased in leaves obtained from C2, especially flavones. However, other secondary metabolites were downregulated in leaves from C2 compared to those detected in leaves from C1, as evidenced by the combination of the variable important in projections (VIP score > 1.3) and the fold-change (FC > 2). The storage of S. minor leaves for 15 days as fresh-cut products did not induce significant changes in the phenolic content and antioxidant capacity, which indicates that the nutraceutical value was maintained. The only difference evidenced during storage was that leaves obtained from C2 showed a lower constitutive content of nutraceutical compounds than leaves obtained from C1; except for chlorophylls and carotenoids. In conclusion, the cut was the main influence on the modulation of secondary metabolites in leaves, and the effects were independent of storage.

Like other members of the Colchicum genus, C. szovitsii subsp. szovitsii is also of medicinal importance in Turkish traditional medicine. However, its biological properties have not been fully investigated. Herein, we focused on the evaluation of the in vitro antioxidant and enzyme inhibitory effects of flower, root and leaf extracts, obtained using different extraction methods. In addition, a comprehensive (poly)-phenolic and alkaloid profiling of the different extracts was undertaken. In this regard, ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) allowed us to putatively annotate 195 polyphenols and 87 alkaloids. The most abundant polyphenols were flavonoids (83 compounds), whilst colchicine and 2-demethylcolchicine were some of the most widespread alkaloids in each extract analyzed. However, our findings showed that C. szovitsii leaf extracts were a superior source of both total polyphenols and total alkaloids (being, on average 24.00 and 2.50 mg/g, respectively). Overall, methanolic leaf extracts showed the highest (p < 0.05) ferric reducing antioxidant power (FRAP) reducing power (on average 109.52 mgTE/g) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging (on average 90.98 mgTE/g). Interestingly, each C. szovitsii methanolic extract was more active than the water extracts when considering enzymatic inhibition such as against tyrosinase, glucosidase, and acetylcholinesterase (AChE). Strong correlations (p < 0.01) were also observed between polyphenols/alkaloids and the biological activities determined. Multivariate statistics based on supervised orthogonal projections to latent structures discriminant analysis (OPLS-DA) allowed for the detection of those compounds most affected by the different extraction methods. Therefore, this is the first detailed evidence showing that C. szovitsii subsp. szovitsii might provide beneficial effects against oxidative stress and the associate

To date, the leading causes of mortality and morbidity worldwide include viral infections, such as Ebola, influenza virus, acquired immunodeficiency syndrome (AIDS), severe acute respiratory syndrome (SARS) and recently COVID-19 disease, caused by the SARS-CoV-2 virus. Currently, we can count on a narrow range of antiviral drugs, especially older generation ones like ribavirin and interferon which are effective against viruses in vitro but can often be ineffective in patients. In addition to these, we have antiviral agents for the treatment of herpes virus, influenza virus, HIV and hepatitis virus. Recently, drugs used in the past especially against ebolavirus, such as remdesivir and favipiravir, have been considered for the treatment of COVID-19 disease. However, even if these drugs represent important tools against viral diseases, they are certainly not sufficient to defend us from the multitude of viruses present in the environment. This represents a huge problem, especially considering the unprecedented global threat due to the advancement of COVID-19, which represents a potential risk to the health and life of millions of people. The demand, therefore, for new and effective antiviral drugs is very high. This review focuses on three fundamental points: (1) presents the main threats to human health, reviewing the most widespread viral diseases in the world, thus describing the scenario caused by the disease in question each time and evaluating the specific therapeutic remedies currently available. (2) It comprehensively describes main phytochemical classes, in particular from plant foods, with proven antiviral activities, the viruses potentially treated with the described phytochemicals. (3) Consideration of the various applications of drug delivery systems in order to improve the bioavailability of these compounds or extracts. A PRISMA flow diagram was used for the inclusion of the works. Taking into consideration the recent dramatic events caused by COVID-19 pa

Background: Watermelon (Citrullus lanatus) represents the largest cultivated member of the Cucurbitaceae family, showing great relevance from the economic point of view and largely consumed around the world accounting for about 7% of the world area dedicated to the production of vegetables, with 67% of the total quantity produced in China. It is characterized by several bioactive compounds, showing different chemical structures, such as carotenoids, xanthophylls, phenolic compounds, citrulline, and unsaturated fatty acids. The good amount of total polyphenols, vitamin C, citrulline and the excellent supply of lycopene, about 40% compared to raw tomatoes, give a measure of the importance of this fruit respect to the other crops considering the beneficial effects of these compounds on human health. Scope and approach: In the last decades, this great interest towards natural bioactive compounds led many researchers to study watermelon as natural source of bioactive compounds, mainly when considering some biological properties, including antioxidant, antimicrobial and anti-inflammatory activities, inhibition or induction of enzymes, inhibition of receptor activities, together with induction/inhibition of gene expression. However, the most of scientific literature on this topic was based on in vitro assays, thus limiting the comprehensive understanding of the real health-promoting outcomes. Key findings and conclusions: Therefore, the aim of this review was to present the up-to-date research carried out on watermelon phytochemicals, showing the most important biological activities reported from both in vitro and in vivo trials. Besides, the potential exploitation of watermelon by-products in the green circular economy and food-waste valorization has been discussed.

Milk is a foodstuff widely consumed around the world originating from a variety of different species, animal management and production systems. In recent years, consumers have placed a much greater emphasis on the authenticity and origin of some food products often willing to pay a premium price for such products that is, for example ‘Grass-Fed Dairy’. Therefore, it is important to establish methods to assess both quality and authentication of milk and dairy products for increased food security and consumer protection. Accordingly, NMR-based, GC–MS-based, and LC–MS-based metabolomics have been established as useful tools in the analysis of dairy products, such as raw and processed milk. This short-review provides an updated and critical overview on the most useful metabolomics-based platforms and the most useful multivariate statistical tools available for metabolomic data interpretation.

Pseudomonas fluorescens is a psychrotrophic species associated with milk spoilage because of its lipolytic and proteolytic activities. Consequently, monitoring P. fluorescens or its antecedent activity in milk is critical to preventing quality defects of the product and minimizing food waste. Therefore, in this study, untargeted metabolomics and peptidomics were used to identify the changes in milk related to P. fluorescens activity by simulating the low-temperature conditions usually found in milk during the cold chain. Both unsupervised and supervised multivariate statistical approaches showed a clear effect caused by the P. fluorescens inoculation on milk samples. Our results showed that the levels of phosphatidylglycerophosphates and glycerophospholipids were directly related to the level of contamination. In addition, our metabolomic approach allowed us to detect lipid and protein degradation products that were directly correlated with the degradative metabolism of P. fluorescens. Peptidomics corroborated the proteolytic propensity of P. fluorescens-contaminated milk, but with lower sensitivity. The results obtained from this study provide insights into the alterations related to P. fluorescens 39 contamination, both pre and post heat treatment. This approach could represent a potential tool to retrospectively understand the actual quality of milk under cold chain storage conditions, either before or after heat treatments.

Pumpkin is considered a healthy and functional food. The consumption of pumpkins and pumpkin-based foods has been shown to confer several beneficial effects on human health due to their antioxidant capacity and terpenoid content. Consequently, this study aimed to characterize the in vitro antioxidant capacity (using FRAP and ABTS assays), terpenoid profile (using an untar-geted lipidomics approach via high-resolution UHPLC-Orbitrap mass spectrometry), and carote-noid content (by HPLC-DAD) in pumpkin fruit pulp from accessions differing for species (11 Cu-curbita maxima and 9 Cucurbita moschata), cultivar, and origin, belonging to a Serbian breeding col-lection. These accessions are candidates for inclusion within programs intended to improve pumpkin fruit quality. The results obtained in this work allowed us to highlight the best marker com-pounds, discriminating both the region of accession collection or breeding (“origin”) and the plant species. Furthermore, our findings have helped to identify the most suitable antioxidant-rich varieties to select for national breeding programs for improving human health. These findings provide valuable information to the overall current understanding of the potential health benefits of pumpkins and the discriminant triterpenoids underlying the C. maxima and C. moschata accessions investigated here, which include those of Serbian and non-Serbian origin.

In this work, the major changes in extra-virgin olive oil (EVOO) composition during cooking were assessed. A foodomics approach based on both metabolomics and lipidomics was used to evaluate the impact of six different cooking techniques, three traditional and three more innovative (Crock-pot®, Roner® and Gastrovac®), and the effect of temperature and cooking time. The lipophilic and hydrophilic fractions of EVOO that underwent different cooking processes were characterized by untargeted high-resolution mass spectrometry approaches. Multivariate statistics were used to unravel the differences in chemical signatures. The different cooking methods resulted in broadly different phytochemical profiles, arising from thermally driven reactions accounting for hydrolysis, synthesis, and oxidation processes. The innovative cooking techniques marginally altered the phytochemical profile of EVOO, whereas sauteing was the cooking method determining the most distinctive profile. Conventional cooking methods (oven, pan-frying, and deep-frying) produced more oxidation products (epoxy- and hydroxy-derivatives of lipids) and markedly induced degradation processes.

Moringa (Moringa oleifera Lam.), the so-called ‘miracle tree’, has emerged as an excellent source of nutrients and functional components also in food applications. In this regard, Moringa is worldwide cultivated for its nutritional properties and for a variety of end-uses. The different portions of this plant (leaves, pods, seeds) have been widely studied for their antioxidant, anti-inflammatory, and antihyperglycemic activities. Most of the functional activities reported in the literature are strictly related to its high content of polyphenols, bioactive peptides, carotenoids, and glucosinolates. Besides, a great interest is emerging towards the exploitation of Moringa as a value-added ingredient for both bakery and meat products. On these bases, considering the more recent literature about M. oleifera plant, this short review aims to highlight the different biological and potential food applications of M. oleifera parts and/or extracts as a source of bioactive compounds able to provide health-promoting properties.

The chemical composition of milk can be significantly affected by different factors across the dairy supply chain, including primary production practices. Among the latter, the feeding system could drive the nutritional value and technological properties of milk and dairy products. Therefore, in this work, a combined foodomics approach based on both untargeted metabolomics and metagenomics was used to shed light onto the impact of feeding systems (i.e., hay vs. a mixed ration based on hay and fresh forage) on the chemical profile of raw milk for the production of hard cheese. In particular, ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF) was used to investigate the chemical profile of raw milk (n = 46) collected from dairy herds located in the Po River Valley (Italy) and considering different feeding systems. Overall, a total of 3320 molecular features were putatively annotated across samples, corresponding to 734 unique compound structures, with significant differences (p < 0.05) between the two feeding regimens under investigation. Additionally, supervised multivariate statistics following metabolomics-based analysis allowed us to clearly discriminate raw milk samples according to the feeding systems, also extrapolating the most discriminant metabolites. Interestingly, 10 compounds were able to strongly explain the differences as imposed by the addition of forage in the cows’ diet, being mainly glycerophospholipids (i.e., lysophosphatidylethanolamines, lysophosphatidylcholines, and phosphatidylcholines), followed by 5-(30,40-Dihydroxyphenyl)-gamma-valerolactone-40-O-glucuronide, 5a-androstan-3a,17b-diol disulfuric acid, and N-stearoyl glycine. The markers identified included both feed-derived (such as phenolic metabolites) and animal-derived compounds (such as lipids and derivatives). Finally, although characterized by a lower prediction ability, the metagenomic profile was found to be significantly correlat

The aim of this work was to investigate the effect of meteorological conditions on resveratrol concentration of red wines produced in Piacenza viticultural region (Italy). In this regard, six representative estates producing Colli Piacentini Gutturnio DOC (a blend of V. vinifera L. cvs. Barbera and Croatina) vintage wines were analysed for trans- and cis-resveratrol over an 8-year period (1998-2005). Grapes were taken from the same vineyard in each estate by using the same enological practices over the entire investigated period. The meteorological conditions corresponding to the production areas were recorded, and bioclimatic indices were calculated as well. Overall, cis-resveratrol concentration was negatively correlated to Huglin index and August mean temperature, whilst positive correlation coefficients were found when considering the Selianinov index and the rainfall of September.

In this work, a metabolomic approach based on ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF) has been applied to investigate the untargeted phenolic profiles of wines from different growing areas, being Val Tidone, Val Nure and Val d'Arda, three valleys located in the province of Piacenza (Emilia Romagna, Italy). In this regard, eight "Gutturnio" DOC wines (both sparkling and still) from the same vintage (2016) were compared in order to highlight the impact of terroir on their phytochemical composition. At the same time, two white wines from the same vintage (2017) were analysed, being "Ortrugo" DOC and "Malvasia di Candia aromatica" DOC. Besides, a sensory evaluation was done in order to underline possible correlations between chemical and climatic data, as related to each growing area. Overall, interesting results were obtained by using UHPLC-ESI-QTOF mass spectrometry; white "Malvasia" and "Ortrugo" wines were very abundant in phenolic acids, being 107.36 and 98.36 mg/L, respectively, while among the "Gutturnio" wines, the highest content were recorded in still Gutturnio wines from Val Tidone and Val d'Arda (i.e., 389.92 and 388.18 mg/L, respectively). Unsupervised multivariate statistics (hierarchical clustering and principal component analysis) following metabolomics-based data allowed to clearly discriminate the different samples according to the corresponding growing-areas. Interestingly, the most discriminant compounds allowing sample grouping belonged to phenolic acids (such as isomeric forms of diferuloylquinic acid) and alkylphenols (such as 5-Heptadecylresorcinol). Besides, the venn diagram analysis revealed 7 common markers belonging to both conditions under investigation (i.e., terroir and winemaking typology). Finally, strong correlations were outlined when considering polyphenol profiles and climatic data, although further ad-hoc studies are extremely needed to confirm this trend.

In this study, an untargeted metabolomics approach based on ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS) was used for investigating changes in chemical profiles of cow milk considering diets based on mycotoxins-contaminated corn silages. For this purpose, 45 milk samples were classified into five clusters according to the corn silage contamination profile, namely (1) low levels of Aspergillus-and Penicillium-mycotoxins; (2) low levels of fumonisins and other Fusarium-mycotoxins; (3) high levels of Aspergillus-mycotoxins; (4) high levels of non-regulated Fusarium-mycotoxins; (5) high levels of fumonisins and their metabolites, and subsequently analyzed by UHPLC-HRMS followed by a multivariate statistical analysis (both unsupervised and supervised statistical approaches). Overall, the milk metabolomic profile highlighted potential correlations between the quality of contaminated corn silages (as part of the total mixed ration) and milk composition. Metabolomics allowed to identify 628 significant milk metabolites as affected by the five levels of corn silage contamination considered, with amino acids and peptides showing the highest metabolite set enrichment (134 compounds). Additionally, 78 metabolites were selected as the best discriminant of the prediction model built, possessing a variable importance in projection score >1.2. The average Log Fold-Change variations of the discriminant metabolites provided evidence that sphingolipids, together with purine and pyrimidine-derived metabolites were the most affected chemical classes. Also, metabolomics revealed a significant accumulation of oxidized glutathione in milk samples belonging to the silage cluster contaminated by emerging Aspergillus toxins, likely involved in the oxidative imbalance. These preliminary findings provide new insights into the potential role of milk metabolomics to provide chemical indicators of mycotoxins-contaminated corn silage fe

In this work, cold-dry fermented nitrate-free sausages were produced by adopting hurdle technologies. The strategy consists of using lactic acid bacteria (LAB) growing at low temperatures, the addition of lactic and ascorbic acids, and low temperatures during the ripening-drying process (35 days at different RH values). Four experimental conditions (B and B1 with 0.2% ascorbic acid, R and R1 with 0.3% lactic acid – B1 and R1 with lower RH during ripening) were compared with a control (N; traditionally manufactured, 80 mg/kg of nitrates and nitrites). Overall, sample R underwent a pH change from 5.22 (t0) to 5.06 (t14). Besides, all samples displayed aw values ≤ 0.92 at t22, thus reaching values < 0.895 at t35. Interestingly, the lower RH values (65–80%) during ripening determined a case hardening, significantly affecting colour parameters. Metabolomics coupled with TBARS assay showed few differences when comparing B and R with the control, whereas lipid oxidation markers (18-hydroxyoleate, LogFC = 0.83–16-Hydroxy-hexadecanoic acid, LogFC = 0.77) and MDA increased in B1 and R1. Overall, RH values revealed to be critical parameters, to be accurately defined to avoid undesired phenomena when hurdle technologies are used. Nonetheless, our findings support the manufacturing of nitrate-free salami by unconventional technologies.

Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) is a severe multisystemic disease characterized by immunological abnormalities and dysfunction of energy metabolism. Recent evidences suggest strong correlations between dysbiosis and pathological condition. The present research explored the composition of the intestinal and oral microbiota in CFS/ME patients as compared to healthy controls. The fecal metabolomic profile of a subgroup of CFS/ME patients was also compared with the one of healthy controls. The fecal and salivary bacterial composition in CFS/ME patients was investigated by Illumina sequencing of 16S rRNA gene amplicons. The metabolomic analysis was performed by an UHPLC-MS. The fecal microbiota of CFS/ME patients showed a reduction of Lachnospiraceae, particularly Anaerostipes, and an increased abundance of genera Bacteroides and Phascolarctobacterium compared to the non-CFS/ME groups. The oral microbiota of CFS/ME patients showed an increase of Rothia dentocariosa. The fecal metabolomic profile of CFS/ME patients revealed high levels of glutamic acid and argininosuccinic acid, together with a decrease of alpha-tocopherol. Our results reveal microbial signatures of dysbiosis in the intestinal microbiota of CFS/ME patients. Further studies are needed to better understand if the microbial composition changes are cause or consequence of the onset of CFS/ME and if they are related to any of the several secondary symptoms.

In this study, four Brassicaceae microgreens species, namely kale, red cabbage, kohlrabi, and radish, were evaluated for their phytochemical compositions using spectrophotometric assays and untargeted metabolomics before and after in vitro gastrointestinal digestion. According to the in vitro spectrophotometric results, significant amounts of phenolics could be detected in each studied species, thus supporting the total antioxidant capacities recorded. Overall, metabolomics allowed annotating a total of 470 phytochemicals across the four Brassicaceae microgreens, either fresh or digested. Among polyphenols, flavonoids were the most represented class (180 compounds, including anthocyanins, flavones, flavonols, and other flavonoids), followed by phenolic acids (68 compounds, mainly hydroxycinnamic and hydroxybenzoic acids), non-flavonoid or phenolic acid-based structures (i.e., alkyl- and alkylmethoxy-phenols and tyrosol derivatives), and lignans. Also, 22 glucosinolates were annotated, including gluconapin glucoraphanin, glucobrassicin, and 4-hydroxyglucobrassicin. Noteworthy, significant differences could be observed in terms of bioaccessibility as a function of the phenolic class and the species considered. Overall, lignans exhibited the highest bioaccessibility values (14%), followed by tyrosol derivatives and flavonoids (on average, 9% and 8%, respectively). However, differences could be evidenced as a function of the species, with red cabbage having comparatively lower bioaccessibility values irrespective of the chemical class of bioactive considered. Similarly, bioaccessibility of glucosinolates significantly differed across species, ranging from 2% in kale to 43% in kohlrabi microgreens.

Gluten-free (GF) biscuits were prepared by replacing part of a GF flour mix (GFM) with 0, 15, 30 and 45 g/100 g (total flour) with a novel resistant starch-rich ingredient obtained from annealed white sorghum starch (RSWS). The chemical composition, physical characteristics, in vitro starch digestion and sensory evaluation of biscuits, were considered. The chemical composition of samples was influenced by the addition of the RSWS. The highest total dietary fibre and RS contents (p < 0.05) were measured in 45-RSWS biscuits. The starch hydrolysis index values decreased when the level of RSWS increased in the composite. With regard to quality parameters, the use of RSWS influenced the hardness of the biscuits, and the highest value obtained for 45-RSWS. Some of the selected sensory attributes, along with the overall acceptability score, were negatively influenced by the RSWS addition, even if all remained above the limit of acceptability. The use of RSWS in GF biscuit formulation can contribute towards the creation of food products likely having slowly digestible starch properties, and this can be achieved without drastically compromising on the quality and sensory attributes.

Oat (Avena sativa L.) is widely appreciated for its beneficial properties for human health, which have led to the introduction of more food products on the market, including oat beverages. The fibre components found in the oat are recognized for their beneficial effects, despite other bioactive compounds with healthy properties being present. This work aimed to evaluate the metabolites profile of a commercial oat beverage, either fermented with lactic bacteria or not, following in vitro gastro-intestinal digestion. UHPLC-QTOF untargeted metabolomics allowed investigation of the bioaccessibility of health-related metabolites from the oat beverage at the intestinal level. The results identified flavonoids, phenolic acids (avenanthramides), amino acids and steroids as the major classes of compounds. In particular, after in vitro digestion, amino acids, peptides, and phenolic acids showed the highest increases. The co-fermentation of oat milk by Lactobacillus spp. and Bifidobacterium spp. strains decreased the levels of both lignans and phytic acid, while increased the levels of some polyphenols like avenanthramides. Furthermore, fermentation by microorganisms increased the bioaccessibility of specific amino acids, vitamins, and polyphenols (flavonoids and phenolic acids). Interestingly, despite lacking a significant part of beta-glucans, the HPAEC-PAD profiling of our oat beverage evidenced that the fermentation process did not alter the oligosaccharides profile, thus preserving its prebiotic potential. The phytochemical profile of oat milk was shown to have a functional potential. Nonetheless, the fermentation by bacterial strains changed the profile of metabolites during in vitro digestion, thus offering an interesting option in the future development of cereal-based beverages.

Breads were prepared by substituting common wheat flour with 0 (GP0), 5 (GP5) and 10 (GP10) g/100 g (w/w) of grape pomace powder (GPP) and were analyzed for the phenolic profile bioaccessibility as well as the in vitro starch digestion during simulated digestion. The free and bound phenolic composition of native GPP and resulting breads were profiled using ultra-high-performance chromatography-quadrupole-time-of-flight (UHPLC-QTOF). The raw GPP was characterized by 190 polyphenols with the anthocyanins representing the most abundant class, ac-counting for 11.60 mg/g of cyanidin equivalents. Regarding the fortified bread, the greatest (p < 0.05) content in phenolic compounds was recorded for the GP10 sample (considering both bound and free fractions) being 127.76 mg/100 g dry matter (DM), followed by the GP5 (106.96 mg/100 g DM), and GP0 (63.76 mg/100 g DM). The use of GPP determined an increase of anthocyanins (considered the markers of the GPP inclusion), recording 20.98 mg/100 g DM in GP5 and 35.82 mg/100 g DM in GP10. The bioaccessibility of anthocyanins increased in both GP5 and GP10 breads when moving from the gastric to the small intestine in vitro digestion phase with an average value of 24%. Both the starch hydrolysis and the predicted glycemic index decreased with the progressive inclusion of GPP in bread. Present findings showed that GPP in bread could promote an antioxidant environment in the digestive tract and influence the in vitro starch digestion.

Eriobotrya japonica, commonly known as loquat, has been used traditionally for the treatment of different diseases. Herein, untargeted profiling based on ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) was used to depict the phytochemical profile of loquat roots, leaves, stems, seeds, and fruits. This allowed the tentative annotation of 349 compounds, representing different phytochemical classes that included flavonoids, phenolic acids, lignans, stilbenes, and terpenoids. Among others, low molecular weight phenolics (tyrosol derivatives) and terpenoids were the most abundant phytochemicals. After that, in vitro antioxidant and enzyme inhibition assays were applied to investigate the biological activity of the different organs of Eriobotrya japonica. Roots of E. japonica exhibited the highest antioxidant capacity, showing 181.88, 275.48, 325.18, 169.74 mg Trolox equivalent (TE)/g in DPPH, ABTS, CUPRAC, and FRAP assays, respectively. Furthermore, the root extract of E. japonica strongly inhibited butyryl cholinesterase (3.64 mg galantamine equivalent (GALAE)/g), whereas leaves, stems, seeds, and fruits showed comparable inhibition of both acetyl and butyryl cholinesterases. All the investigated organs of E. japonica exhibited in vitro tyrosinase inhibition (57.27–71.61 mg Kojic Acid Equivalent (KAE)/g). Our findings suggest a potential food and pharmaceutical exploitation of different organs of E. japonica (mainly roots) in terms of enrichment with health-promoting phenolics and triterpenes.

Low-molecular-weight compounds in milk are of interest from both nutritional and technological perspectives. To analyse potential milk metabolites, acetonitrile extraction followed by ethyl chloroformate derivatisation was developed. A GC/MS (SIM) method was then applied to different animal milk samples (i.e., buffalo, bovine, and donkey) and processed milk samples (i.e., pasteurised and dried). The optimised extraction-derivatisation method is rapid and more comprehensive when compared with the other published methods. The supervised orthogonal projection to latent structure discriminant analysis (OPLS-DA) based on the amino acid profile showed clear discrimination as a function of animal origin and milk processing. In this regard, variable importance in projection (VIP) analysis following an OPLS-DA prediction model showed that aspartic acid and asparagine (VIP scores = 1.26 and 1.19, respectively) were the best markers of the milk origin, whilst proline and glycine (VIP scores = 1.30 and 1.28, respectively) mainly discriminated samples according to different processing conditions.

In this preliminary study, ultra-high-pressure liquid chromatography (UHPLC) coupled to quadrupole-time-of-flight mass spectrometry (QTOF) metabolomics followed by multivariate statistics was applied to discriminate nine extra-virgin olive oil (EVOO) cultivars according to their phenolic and sterolic fingerprints. The same approach was then used to discriminate EVOO samples from different geographical origins, namely six blends representative of the main growing regions in Italy (i.e., Sicily, Puglia, Umbria, Liguria, Lombardy and Tuscany). This approach allowed to putatively identify >1000 compounds, considering both polyphenols and sterols. The unsupervised hierarchical cluster analysis (HCA) and the orthogonal projections to latent structures discriminant analysis (OPLS-DA) discriminated EVOO samples according to both cultivar and geographical origin. In particular, flavonoids (i.e., anthocyanins and flavonols), hydroxycinnamic acids and cholesterol derivatives were found to be the most representative classes of compounds discriminating EVOO samples according to the two parameters (cultivar or geographical origin) selected. However, the following Venn analysis allowed to point out the discriminant markers being exclusive for cultivar or origin discrimination. In this regard, only the 9.6% of phenolics and 13.6% of sterols were in common, thus indicating that several of these discriminant compounds were exclusive of a single condition. Indeed, considering that most of the commercial EVOOs are blended, the contribution of both the cultivars used and the geographical origin must be taken into account.

We evaluated the impact of dietary supplementation with a commercially available smectite clay (TOXO® MX, Trouw Nutrition, Amersfoort, The Netherlands), that binds to aflatoxins (AFs), on the performance and health status of multiparous lactating Holstein dairy cows that received dietary AFB1 (the main AF). The carry-over of AFB1 was determined by measuring AFM1 (the main metabolite) in dairy milk. Performance values, blood markers, and liver inflammatory markers were also measured. Nine multiparous mid-lactation Holstein cows (parity: 2.67 ± 0.86; days in milk: 91 ± 15 days; milk yield: 40.4 ± 2.7 kg/cow/day) were assigned to one of three treatments in a 3 periods × 3 treatments Latin square design (n = 3). In particular, three cows each received the CTR-0 diet (total mixed ration (TMR) with normal corn meals), the CTR-AFLA diet (CTR-0 diet with 17.53 ± 6.55 µg/kg DM AFBI), or the TRT diet (CTR-AFLA diet with 100 ± 1 g/cow/day of smectite clay). The AFB1 level was 0.63 ± 0.50 µg/kg DM in the CTR-0 diet, 2.28 ± 1.42 µg/kg DM in the CTR-AFLA diet, and 2.13 ± 1.11 µg/kg DM in the TRT diet. The experiment consisted of an adaptation period (21 days) and three 17-day experimental periods, each consisting of a 10-day intoxication period and 7-day clearance period. Data were analyzed using the MIXED procedure of SAS (SAS Inst. Inc., Cary, NC, USA) with or without repeated measurements. Overall, the addition of AFB1 reduced the DM intake, but the groups had no significant differences in milk yields. The highest feed efficiency was in the TRT group. Measurement of AFM1 in milk indicated a “plateau” period, from day 4 to day 10 of the intoxication period, when the AFM1 level exceeded the guidelines of the European Union. The commercial smectite clay reduced milk AFM1 concentration by 64.8% and reduced the carry-over by 47.0%. The CTR-0 and TRT groups had similar carry-over levels of AFM1, although the absolute concentrations differed. The groups had no significant differences