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A comprehensive evaluation of the attributes that determine caviar quality, including sturgeon species and processing methods, is still lacking in the scientific literature. In this study, eight types of caviar, obtained from six different sturgeon species and produced using either salt or a combination of salt and sodium tetraborate (E285) with pasteurization, were characterized through untargeted metabolomics, sensory analysis, and texture evaluation. The results showed that Huso huso caviar was rich in gamma-glutamyl peptides (contributing to a kokumi taste), while the other sturgeon species were primarily distinguished by lipids (mainly glycerophospholipids) and nucleotides (such as AMP, inosinic acid, and other metabolites driving umami taste). Moreover, untargeted metabolomics revealed the technological effects of salting and pasteurization. Sensory analysis identified crunchiness, color homogeneity, and darkness as the main traits differentiating the products. Butter, hazelnut, and briny flavours were also key characteristics of the caviar., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This research can be considered as the first complete survey for the valorization of new olive genotypes cultivated in the South-East of Tunisia as well as their oils. The study aimed to characterize the phytochemical composition of virgin olive oil produced from two olive cultivars, namely Nourgou and Gousalani. The pomological characterization of fruits, the quality criteria and the phytochemical profile were quantified. Additionally, antioxidant activity was evaluated using Ferric reducing antioxidant power (FRAP) and Oxygen radical absorbance capacity (ORAC) tests to also obtain a bioactive characterization of these monovarietal olive oils. The obtained results revealed that the analyzed olive oils samples can be classified into Extra Virgin category (EVOO) according to the regulated physicochemical characteristics. Our findings showed a significant variability in the chemical parameters of the analyzed EVOO likely associated with the genetic potential, mainly for chlorophylls contents (1.37-1.64 mg/kg), in carotenoids pigments (3.97-10.86 mg/kg), in α-tocopherol (175.59-186.87 mg/kg), in sterols (1036.4-1931.4 mg/kg) in oleic acid (65.33-68.73%), in palmitic acid (C16:0) (13.32-17.48%), in linoleic acid (C18:2) (11.06-13.47%). Additionally, the HPLC-MS/MS analysis showed that the two EVOOs analyzed contained appreciable amounts of total polyphenols, ranging from 348.03 up to 516.16 mg/kg, in Nourgou and Gousalani oils, respectively. Regarding the individual phenolic compounds, the EVOO samples were mainly characterized by phenolic alcohols, phenolic acids, secoiridoids, verbascoside, flavonoids and phenolic aldehydes. The prevalent simple phenolics detected were secoiridoids with the dominance of the oleuropein aglycone in Gousalani oil. In addition, findings from in vitro antioxidant assays (FRAP and ORAC) revealed that the two studied oils possessed a powerful antiradical activity and a good reducing power capacity. In conclusion, these new EVOOs exhibited a superior quality compared to other Tunisian varieties, considering their antiradical activity and reducing power capacity.

Utilizing different chymosin and pepsin ratios in cheesemaking may represent a potential strategy to shape the sensory profile of hard cheeses. This study investigated the impact of rennet with varying chymosin and pepsin ratios on the chemical profile and sensory attributes of Grana Padano PDO cheese at different ripening times (10 to 20 months). The research involved the analysis of hard cheese manufactured with distinct calf chymosin percentages (99 %, 95 %, and 83 %), exploiting sensory analyses and untargeted metabolomics to identify marker compounds correlating with specific sensory traits. The results demonstrated that varying the rennet composition significantly affected sensory profile; in particular, the rennet made by 83 % chymosin and 17 % pepsin generated a more complex sensory profile starting from 12 months. AMOPLS and ASCA analysis on untargeted metabolomics signatures revealed that ripening time was the only significant factor when compared with rennet type and the interaction ripening x rennet. Finally, at more advanced ripening times, 3-methylbutanoic acid and homoethone were significantly up-accumulated in cheese samples manufactured with higher pepsin percentages, likely explaining sensory outcomes. This study provides valuable insights into using rennet to tailor the sensory qualities of hard cheeses, underscoring the importance of enzyme selection in cheese manufacturing to drive innovation in the dairy industry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The chemical composition of Parmigiano Reggiano (PR) hard cheese can be significantly affected by different factors across the dairy supply chain, including ripening, altimetric zone, and rind inclusion levels in grated hard cheeses. The present study proposes an untargeted metabolomics approach combined with machine learning chemometrics to evaluate the combined effect of these three critical parameters. Specifically, ripening was found to exert a pivotal role in defining the signature of PR cheeses, with amino acids and lipid derivatives that exhibited their role as key discriminant compounds. In parallel, a random forest classifier was used to predict the rind inclusion levels (> 18%) in grated cheeses and to authenticate the specific effect of altimetry dairy production, achieving a high prediction ability in both model performances (i.e., ∼60% and > 90%, respectively). Overall, these results open a novel perspective to identifying quality and authenticity markers metabolites in cheese., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Oleuropein (OLE), a phenolic compound particularly abundant in the olive leaves, has been reported to have beneficial activities against colorectal cancer (CRC). In vitro studies suggested that these latter could be due to a modulation of the intestinal microbiota. Aiming to evaluate if OLE could affect the intestinal microbiota and the plasma metabolome, an antioxidant oleuropein-rich leaf extract (ORLE) was administered for one week to PIRC rats (F344/NTac- Apc am1137 ), a genetic model mimicking CRC. ORLE treatment significantly modulated the gut microbiota composition. Plasma metabolomic profiles revealed a significant predictive ability for amino acids, medium-chain fatty acids, and aldehydes. Pathway analysis revealed a significant decrease in phosphatidylcholine accumulation (LogFC = -1.67) in PIRC rats. These results suggest a significant effect of ORLE administration on faecal microbiota profiles and plasma metabolomes, thereby offering new omics-based insights into its protective role in CRC progression.

Mycotoxins are well-known secondary metabolites produced by several fungi that grow and occur in different crops during both pre-harvest and post-harvest conditions. The contamination and occurrence of mycotoxins currently represent some of the major issues in the entire agri-food system. The quantification of mycotoxins in different feeds and foodstuffs is extremely difficult because of the low concentration ranges; therefore, both sample collection and preparation are essential to providing accurate detection and reliable quantification. Currently, several analytical methods are available for the detection of mycotoxins in both feed and food products, and liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) represents the most reliable instrumental approach. In particular, the fast development of high-throughput methods has made it possible to screen and analyze, in the same analytical run and with high accuracy, multiple mycotoxins, such as those regulated, masked, or modified, and emerging ones. Therefore, the aim of this review is to provide an overview of the state of the art of mycotoxins occurrence, health-related concerns, and analyses, discussing the need to perform multi-screening approaches combined with omics technologies to simultaneously analyze several mycotoxins in different feed and food matrices. This approach is expected to provide more comprehensive information about the profile and distribution of emerging mycotoxins, thus enhancing the understanding of their co-occurrence and impact on the entire production chain.

Mealworm (MWP), migratory locust (LP), and house cricket (CP) are novel foods recently authorized by the European Commission. This work tested their powders as meat extenders at 5% inclusion in beef burgers. Insect powders were abundant in phenolics, recording the highest values in LP (1184.9 μg/g). The sensory analysis highlighted a higher visual and olfactory acceptability for MWP-burgers, followed by CP- and LP-burgers, whereas the texture of cooked burgers remained unaffected. Following pan-cooking, MWP-burgers and control exhibited comparable chemical profiles, while a significant down-accumulation of the heterocyclic amine 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline was observed in CP-burgers. In vitro gastrointestinal digestion highlighted metabolomic trends like control for MWP- and LP-burgers. In contrast, a reduced accumulation of lipids and increased content of dipeptides like glutaminylarginine (possibly acting as enzyme modulators) was observed for the CP-burgers., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

A commercial olive leaf extract (OL), effective against Salmonella enterica , Escherichia coli , Listeria monocytogenes , and Staphylococcus aureus , was added to three different coating formulations (methylcellulose, MC; chitosan, CT; and alginate, ALG) to produce active polylactic acid (PLA) coated films. Evaluation of these coated PLA films revealed significant inhibition of S. aureus growth, particularly with the MC and CT formulations exhibiting the highest inhibition rates (99.7%). The coated films were then tested for food contact compatibility with three food simulants (A: 10% ethanol; B: 3% acetic acid; D2: olive oil), selected to assess their suitability for pre-cut hams and ready-to-eat vegetables in relation to overall migration. However, coated films with active functions exhibited migration values in simulants A and B above legal limits, while promising results were obtained for simulant D2, highlighting the need to deeply investigate these coatings' impact on a real food system. Untargeted metabolomics revealed that the type of coating influenced the selective release of certain phenolic classes based on the food simulant tested. The Oxitest analysis of simulant D2 demonstrated that the MC and ALG-coated PLA films slightly slowed down the oxidation of this food simulant, which is an edible vegetable oil.

Nitrates and nitrites, which are synthetic additives, are traditionally used as curing agents in meat-based products. These synthetic additives are employed in the preparation of fermented meat foods to improve quality characteristics and microbiological safety, develop distinct flavours and red-colour stability, and counteract lipid oxidation. Nitrites also display significant bacteriostatic and bactericidal action against spoilage microorganisms and foodborne pathogens (such as Clostridium botulinum and Listeria monocytogenes ). However, meat curing is currently under scrutiny because of its links to cardiovascular diseases and colorectal cancer. Based on the current literature, this review provides recent scientific evidence on the potential utilisation of coagulase-negative staphylococci (CNS) as nitrate and nitrite substitutes in meat-based foods. Indeed, CNS are reported to reproduce the characteristic red pigmentation and maintain the typical high-quality traits of cured-meats, thanks to their arginine degradation pathway, thus providing the nitrite-related desirable attributes in cured meat. The alternative strategy, still based on the NOS pathway, consisting of supplementing meat with arginine to release nitric oxide (NO) and obtain a meat characterised by the desired pinkish-red colour, is also reviewed. Exploiting NOS-positive CNS strains seems particularly challenging because of CNS technological adaptation and the oxygen dependency of the NOS reaction; however, this exploitation could represent a turning point in replacing nitrates and nitrites in meat foods., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Mealworm, migratory locust, and house cricket have recently been recognized by the European Commission as novel foods, thus being suitable in different food applications. In this work, we tested their powders as meat extenders at 5% ( w / w ) inclusion in beef burgers, considering their ability to vehicle phenolic compounds during simulated in vitro static gastrointestinal digestion (INFOGEST). Insect powders were abundant in different phenolic classes, recording the highest values in locust (LP; 314.69 mg/kg), followed by cricket (CP; 113.3 mg/kg) and mealworm (MWP; 51.9 mg/kg). Following a pan-cooking process, LP burgers were confirmed as the best source of phenolics, with a marked abundance of flavonoids and phenolic acids. Interestingly, the insect powders were found to affect the in vitro gastrointestinal bioaccessibility of phenolic compounds when compared with the CTR burger, likely promoted by the interactions between the phenolic compounds and proteins characterizing the tested insect powders. Among the most discriminant phenolic metabolites at the gastrointestinal level, we found several phenolic acids (mainly hydroxycinnamics), recording the highest content for the digested CP-containing burgers. Finally, stilbenes showed significant correlation values at the intestinal level with both antioxidant and enzymatic activities, while total flavonoids were the most correlated with the inhibition of acetylcholinesterase. Taken together, our preliminary findings demonstrated that insect powders added to beef burgers can promote the bioaccessibility and potential bioavailability of phenolics in the distal tracts of the intestine.

The Caco-2 cellular permeability of phenolic aqueous extracts from blackcurrant press cake (BC), Norway spruce bark (NS), scots pine bark (SP), and sea buckthorn leaves (SB) was evaluated by combining high-resolution mass spectrometry and atomic force microscopy. Besides, Caco-2 and HepG2 cells allowed the study of intracellular oxidative stress assessed in both apical and basolateral domains. Overall, BC and NS showed the highest total phenolic contents, 4.38 and 3.76 µg/mL, respectively. Multivariate statistics discriminated NS and BC from SP and SB extracts because of their phenolic profile. Polyphenols were classified as highly permeable, thus suggesting their potentially high bioavailability through the gastrointestinal tract. All the phenolic subclasses showed efflux ratio values < 1, except for BC flavonols, flavan-3-ols, and stilbenes. Regarding cellular damage, NS and BC extracts, when acting on the basolateral cellular side, caused epithelial leakage and morphological shape cell damage on Caco-2 cells associated with ROS production., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Introduction: Feed additives represents a valid tool in animal nutrition to improve animal performance and livestock productivity under a sustainable perspective; however, there is a paucity of information about their comprehensive metabolomic and bioactive profiles., Objective: In this study, we tested the ability of an untargeted metabolomics approach to discriminate nine commercial feed additives and unique blends of botanical extracts used in both ruminant and non-ruminant nutrition, according to their phytochemical profiles and different in vitro bioactive properties., Methods: An ultra-high-performance liquid chromatography coupled with Orbitrap mass spectrometry and multivariate statistics were combined to search for potential markers, in order to better discriminate the different commercial samples., Results: Several phytochemicals were identified, namely alkaloids, phenolics, organosulfurs, and terpenoids. The polyherbal formulation Zigbir was the best source of phytochemicals, accounting for a cumulative total content of phytochemicals equal to 3.03 mg Eq./g, being particularly abundant in terpenoids, stilbenes, phenolic acids, and small-molecular-weight phenolics. Multivariate statistics allowed to group the different products in 2 bioactive subclusters. The diterpenoid andrographolide recorded the highest abundance in Zigbir and Sangrovit. The most predictive biomarkers were: piperine, isoquercitrin, 6-methylthiohexyldesulfoglucosinolate, 6-methylumbelliferone, benzoic acid, (+)-(1R,2R)-1,2-diphenylethane-1,2-diol, and piperitenone. Flavonoids were highly correlated with both in vitro antioxidant and enzyme inhibition assays., Conclusions: Our findings provide new insights into the comprehensive phytochemical composition of commercial feed additives and blend of botanical extracts used for both ruminant and non-ruminant nutrition. A great importance of polyphenols in relation to the biological activities was detected., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Hard cheeses may occasionally show a brown discolouration during ripening due to multifactorial phenomena that involve bacteria and give rise to pyrazines arising from methylglyoxal. The present work aimed at developing a novel approach to investigate the role of natural starters in browning. To this object, 11 strains of L. helveticus were incubated in a medium containing 10 % rennet casein dissolved in whey, and then growth was monitored by measuring pH and number of genomes/mL. Browning was assessed through CIELab analysis, methylglyoxal production was determined by targeted mass spectrometry, and untargeted metabolomics was used to extrapolate marker compounds associated with browning discoloration. The medium allowed the growth of all the strains tested and differences in colour were observed, especially for strain A7 (ΔE* value 15.92 ± 0.27). Noteworthy, this strain was also the higher producer of methylglyoxal (2.44 µg/mL). Metabolomics highlighted pyrazines and β-carboline compounds as markers of browning at 42 °C and 16 °C, respectively. Moreover, multivariate statistics pointed out differences in free amino acids and oligopeptides linked to proteolysis, while 1,2-propanediol and S-Lactoylglutathione suggested specific detoxification route in methylglyoxal-producing strains. Our model allowed detecting differences in browning amid strains, paving the way towards the study of individual L. helveticus strains to identify the variables leading to discoloration or to study the interaction between different strains in natural whey starters., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The impact of duckweed extracts (DEs) on the shelf-life of packaged beef burgers was evaluated through classical assays and untargeted metabolomics. Beef burgers were formulated with an antioxidants-free control (CON), 1 g/kg sodium ascorbate (ASC), and increasing levels of a DEs, namely 1 (DE1), 5 (DE5), and 10 (DE10) g/kg, packaged under modified atmosphere and stored at 4 °C for 19 days. The DEs, abundant in phytochemicals, determined no issues with the hygienic status of the product. DEs modulated the redox status, being ineffective in preserving linolenic acid from peroxidation. However, the oxidation marker 2-nonenoic acid was down-accumulated in the DE10 sample following 19 days of storage, recording a lower glutathione:glutathione disulfide ratio. The accumulation of adipate semialdehyde revealed the inefficiency of DEs in coping with protein oxidation, while DEs prevented the accumulation of biogenic amines. Therefore, this work suggests a potential pro-oxidant role of the formulated DEs., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Author(s).)

This study examined the phytochemical profiles (mainly phenolics, carotenoids, and organosulfur compounds) and biological effects of hydroalcoholic extracts of Allium flavum (AF), a species of the Allium genus commonly known as small yellow onion. Unsupervised and supervised statistical approaches revealed clear differences between extracts prepared with samples collected from different areas of Romania. Overall, the AFFF (AF flowers collected from Făget) extract was the best source of polyphenols, also showing the highest antioxidant capacity evaluated through both in vitro DPPH, FRAP, and TEAC anti-radical scavenging assays and cell-based OxHLIA and TBARS assays. All the tested extracts exhibited α-glucosidase inhibition potential, while only the AFFF extract exhibited anti-lipase inhibitory activity. The phenolic subclasses annotated were positively correlated with the assessed antioxidant and enzyme inhibitory activities. Our findings suggested that A. flavum has bioactive properties worth exploring further, being a potential edible flower with health-promoting implications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

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 ± 0.14 to 9.45 ± 0.06 Log CFU mL -1 and proved capable of producing different vitamin B9 vitamers after 72 h incubation at 30°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 biosynthesis. These isolates appear interesting candidates for further ad-hoc investigation regarding the production of pro-bioactive scotta., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Coronas, Zara, Gallo, Rocchetti, Lapris, Petretto, Zara, Fancello and Mannazzu.)

Nutraceuticals and functional foods are composed of especially complex matrices, with polyphenols, carotenoids, minerals, and vitamins, among others, being the main classes of phytochemicals involved in their bioactivities. Despite their wide use, further investigations are needed to certify the proper release of these phytochemicals into the gastrointestinal medium, where the bioaccessibility assay is one of the most frequently used method. The aim of this review was to gather and describe different methods that can be used to assess the bioaccessibility of nutraceuticals and functional foods, along with the most important factors that can impact this process. The link between simulated digestion testing of phytochemicals and their in vitro bioactivity is also discussed, with a special focus on the potential of developing nutraceuticals and functional foods from simple plant materials. The bioactive potential of certain classes of phytochemicals from nutraceuticals and functional foods is susceptible to different variations during the bioaccessibility assessment, with different factors contributing to this variability, namely the chemical composition and the nature of the matrix. Regardless of the high number of studies, the current methodology fails to assume correlations between bioaccessibility and bioactivity, and the findings of this review indicate a necessity for updated and standardized protocols., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Nicolescu, Babotă, Barros, Rocchetti, Lucini, Tanase, Mocan, Bunea and Crișan.)

Polyphenols are multifaceted bioactive compounds, but little is known about their real impact on human health after consumption. In this work, the phenolic profiling of quebracho, yellow maize, and violet rice extracts was comprehensively investigated, together with the impact of in vitro digestion and colonic fermentation on the bioaccessibility and bioavailability of these phytochemicals. The different matrices showed distinct profiles, potentially influencing in vitro starch digestion under cooking conditions. Furthermore, after the extracts underwent in vitro gastrointestinal digestion and faecal fermentation, phenolics exhibited a differential bioaccessibility trend at every digestion level, with matrix-dependent behaviour. The bioavailability results suggest that polyphenols are metabolised during colonic fermentation, mainly into tyrosols, phenolic acids, and lignans, which are partially absorbed by Caco-2 cells. By combining metabolomics with in vitro cellular methods, this research provides new insights into the fate of these phytochemicals in the gut, yielding comprehensive data on their consumption in food matrices., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

In this study, a rowanberry pomace defatted with supercritical CO 2 (2%-AC), its ethanolic extract (1%-E) and extraction residue (2%-R), were tested in meatball preparation. The meatballs with 1%-E demonstrated the highest in vitro radical scavenging capacity. In the case of 1%-E the pH of meatballs was significantly lower compared to the control sample (P = 0.0132) on the 5-day. The lowest cooking loss was achieved when the meatballs contained mainly fibre-rich 2%-R. The UHPLC method detected 184 metabolites, including strong antioxidants, such as chlorogenic acids, 3',4'-methylenedioxy-5,7-dimethylepicatechin, hyperin, isoquercitrin. The 1%-E was particularly effective against the development of unpleasant off-flavours caused by carbonyl compounds. Consistently, the decrease in lipid oxidation, indicated by reduced 7-dodecenal and 2,4-heptadienal contents, has been observed following the addition of rowanberry extract to meatballs. Metabolomics coupled with conventional quality evaluations provided a deeper understanding of the potential utilization and valorisation of different rowanberry pomace extracts as meat ingredients., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Author(s).)

Blackcurrants are sources of phenolic compounds, such as anthocyanins, possessing strong antioxidant, antimicrobial and antifungal activity. Therefore, the addition of different blackcurrant pomace ingredients may affect the overall meat quality. The actual chemical profile and bioactivities of blackcurrant pomace ingredients may strongly depend on its preparation; for instance, in our study the highest values of the in vitro antioxidant capacity were determined for blackcurrant seeds after supercritical CO 2 extraction. Starting from these background conditions, in this work, we evaluated the ability of three different concentrations (namely 1, 3, and 5% w/w) of blackcurrant (BC) seeds following EtOH/water extraction (BC-AE), before supercritical fluid CO 2 extraction (BC-RS), and after supercritical fluid CO 2 extraction (BC-ASC) to affect different quality parameters of pork meatballs. These latter were stored considering three different time-points, namely 1, 3 and 6 days at 4 °C packed under modified atmosphere (i.e., 70% N 2 and 30% CO 2 ). Untargeted metabolomics allowed to identify several lipid and protein-related oxidation products involved in redox reactions, such as 13-L-hydroperoxylinoleic acid, (12S,13S)-epoxylinolenic acid, 9,10-epoxyoctadecenoic acid, glutathione, glutathione disulfide, L-carnosine, l-ascorbic acid, and tocotrienols. Besides, multivariate statistics applied on the metabolomics dataset confirmed that the chemical profile of meatballs was an exclusive combination of both BC inclusion levels and type of BC-ingredients considered. Our findings showed that the higher the concentration of BC seed ingredients in meatballs, the lower the cooking loss and the higher the fibre content. Also, all the ingredients significantly affected the colour parameters., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Platinum nanoparticles (PtNPs) are being intensively explored as efficient nanozymes due to their biocompatibility coupled with excellent catalytic activities, which make them potential candidates as antimicrobial agents. Their antibacterial efficacy and the precise mechanism of action are, however, still unclear. In this framework, we investigated the oxidative stress response of Salmonella enterica serovar Typhimurium cells when exposed to 5 nm citrate coated PtNPs. Notably, by performing a systematic investigation that combines the use of a knock-out mutant strain 12023 HpxF - with impaired response to ROS (Δ katE Δ katG Δ katN Δ ahpCF Δ tsaA ) and its respective wild-type strain, growth experiments in both aerobic and anaerobic conditions, and untargeted metabolomic profiling, we were able to disclose the involved antibacterial mechanisms. Interestingly, PtNPs exerted their biocidal effect mainly through their oxidase-like properties, though with limited antibacterial activity on the wild-type strain at high particle concentrations and significantly stronger action on the mutant strain, especially in aerobic conditions. The untargeted metabolomic analyses of oxidative stress markers revealed that 12023 HpxF - was not able to cope with PtNPs-based oxidative stress as efficiently as the parental strain. The observed oxidase-induced effects comprise bacterial membrane damage as well as lipid, glutathione and DNA oxidation. On the other hand, in the presence of exogenous bactericidal agents such as hydrogen peroxide, PtNPs display a protective ROS scavenging action, due to their efficient peroxidase mimicking activity. This mechanistic study can contribute to clarifying the mechanisms of PtNPs and their potential applications as antimicrobial agents.

Ripening time is known to drive the chemical and sensory profiles of dry meat products, thus potentially affecting the final quality of the product. Starting from these background conditions, the aim of this work was to shed light, for the first time, on the chemical modifications of a typical Italian PDO meat product-namely, Coppa Piacentina-during ripening, to find correlations between its sensory quality and the biomarker compounds related to the progress of ripening. The ripening time (from 60 to 240 days) was found to deeply modify the chemical composition of this typical meat product, providing potential biomarkers of both oxidative reactions and sensory attributes. The chemical analyses revealed that there is typically a significant decrease in the moisture content during ripening, likely due to increased dehydration. In addition, the fatty acid profile showed that the distribution of polyunsaturated fatty acids significantly ( p < 0.05) decreased during ripening, because of their high susceptibility to oxidation and conversion to intermediate and secondary molecules. An untargeted metabolomics approach, coupled with unsupervised and supervised multivariate statistics, highlighted a significant impact (prediction scores > 1) of lipid oxidation during ripening time, with some metabolites (such as γ -glutamyl-peptides, hydroperoxy-fatty acids, and glutathione) being particularly discriminant in predicting the changes observed. The discriminant metabolites were coherent with the progressive increase of peroxide values determined during the entire ripening period. Finally, the sensory analysis outlined that the highest degree of ripening provided greater color intensity of the lean part, slice firmness, and chewing consistency, with glutathione and γ-glutamyl-glutamic acid establishing the highest number of significant correlations with the sensory attributes evaluated. Taken together, this work highlights the importance and validity of untargeted metabolomics coupled with sensory analysis to investigate the comprehensive chemical and sensory changes to dry meat during ripening.

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 natriuretic and kaliuretic effect after the administration. In terms of antimicrobial activity, E. coli (MIC-0.38 mg/ml), B. cereus (MIC-0.75 mg/ml)), Penicillium funiculosum and P. verrucosum var. cyclopium (MIC-0.19 mg/ml) showed the greater sensitivity to the tested extracts, respectively. UHPLC-HRMS screening showed that the bioactive potential of T. comosus herbal preparations was likely related to the higher amounts of phenolic acids (including rosmarinic acid), flavonoids (mainly flavones and derivatives) and other phenolics (such as different isomers of salvianolic acids) in their composition. The obtained results support the ethnopharmacological evidence regarding the mild diuretic and antibacterial potentials of the endemic wild thyme T. comosus , this study being the first one that assessed the aforementioned bioactivities for this species., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Babotă, Frumuzachi, Nicolescu, Stojković, Soković, Rocchetti, Zhang, Lucini, Crișan, Mocan and Voștinaru.)

In this work, the synergistic/antagonistic impact of glucose and mixed starter cultures, namely Latilactobacillus sakei, Pediococcus pentosaceus, and Staphylococcus xylosus, was evaluated in Italian salami in terms of metabolomics and sensory profiles. As expected, Salami manufactured with 0.5 % glucose exhibited a substantial pH drop, showing values close to 5 at 12 days of ripening. Metabolomics revealed 1841 metabolites, mainly belonging to amino acids, peptides, glycerolipids, and nucleic acids, showing a greater hierarchical role of glucose addition when compared with inoculated microbial starters. Distinct metabolomic fingerprints could be observed across treatments, mainly concerning glutamyl peptides like gamma-glutamyl-glutamate (related to the kokumi taste), biogenic amines (spermine), and lipid oxidation products (i.e., the oxylipin 13S-hydroperoxylinolenic acid). Such differences may drive the differences in sensory profiles recorded among treatments. These findings indicate the need to select ad-hoc starter cultures to improve the safety, quality, and sensory traits of salami., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The present study was designed to evaluate the functional potential of common duckweed ( Lemna minor L.) as a source of bioactive compounds of nutraceutical interest. The untargeted profiling of the bioactive components of common duckweed was carried out through ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS), in parallel with assessing in vitro antioxidant and enzymatic inhibition properties. The optimization of extraction parameters was determined using the response surface methodology (RSM) through a 3-factor central composite design. The process parameters included extraction temperature, % of ethanol, and ultrasound power, while the response variables were the phenolic content (considering each main phenolic class), total glucosinolates, total carotenoids, the antioxidant potential, and enzyme inhibition activities. The results revealed that common duckweed was a rich source of carotenoids and total flavonoids (mainly flavones and flavonols), followed by phenolic acids, low-molecular-weight phenolics, and glucosinolates. Interestingly, the total flavones, total flavonols and total carotenoid equivalents showed the highest and most positive correlation values with the bioactive properties measured. Finally, the combined RSM approach and unsupervised statistics allowed us to point out the pivotal impact of ethanol percentage in the extraction solvent to recover the highest amounts of bioactive compounds efficiently.

In this work, untargeted metabolomics was used to shed light on the impact of different pasture-based diets on the chemical profile of Sarda sheep milk. The study considered 11 dairy sheep farms located in Sardinia, and milk samples were collected in 4 different periods, namely January, March, May, and July 2019, when all sheep had 58, 98, 138, and 178 days in milk, respectively. The animal diet composition was based on the intake of grazed herbage in natural pasture, hay, and concentrate. Overall, the combination of two comprehensive databases on food, namely the Milk Composition Database and Phenol-Explorer, allowed the putative identification of 406 metabolites, with a significant (p < 0.01) enrichment of several metabolite classes, namely amino acids and peptides, monosaccharides, fatty acids, phenylacetic acids, benzoic acids, cinnamic acids, and flavonoids. The multivariate statistical approach based on supervised orthogonal projections to latent structures (OPLS-DA) allowed us to predict the chemical profile of sheep milk samples as a function of the high vs no fresh herbage intake, while the prediction model was not significant when considering both hay and concentrate intake. Among the discriminant markers of the herbage intake, we found five phenolic metabolites (such as hippuric and coumaric acids), together with lutein and cresol (belonging to carotenoids and their metabolites). Additionally, a high discriminant power was outlined for lipid derivatives followed by sugars, amino acids, and peptides. Finally, a pathway analysis revealed that the herbage intake affected mainly five biochemical pathways in milk, namely galactose metabolism, phenylalanine metabolism, alpha-linolenic acid metabolism, linoleic acid metabolism, and aromatic amino acids involved in protein synthesis (namely tyrosine, phenylalanine, and tryptophan).

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 °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.

Lactic Acid Bacteria (LAB) could provide a valid alternative to S. cerevisiae as a starter culture for bakery products, avoiding yeast-related health problems while contributing to the technological and functional properties of bread. In this work, we evaluate the role of certain LABs (Leuconostoc citreum SB6, Weissella cibaria UC4051, Weissella confusa UC4051, and the commercial starter cultures Weissella cibaria, and Leuconostoc mesenteroides) in producing functional compounds (pro-technological, health-promoting, and postbiotic-like molecules). For this purpose, we analysed the genotypic and phenotypic features of strains, and we investigated dough fermentation from microbiological and metabolomics approaches. Results evidenced a clear discrimination between the metabolic activity of baker's yeast and LAB. The most discriminant metabolites derived from proteolysis and lipolysis, such as peptides, amino acids, and fatty acyls. Furthermore, we elucidated the different metabolism of these strains by building gene-metabolite interaction networks that pairwise compared the LAB strains of the same genus. While most of the networks showed a characteristic nucleotide metabolism, only the commercial W. cibaria exhibited an interaction network composed of amino acids and their related genes. In conclusion, our findings reveal that LAB strains under investigation, and particularly the commercial W. cibaria, can enhance the functional properties of bread., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In this work, we studied the impact of encapsulated elderberry extracts as natural meat extenders to preserve both the quality and the oxidative stability of beef burgers. In particular, the comprehensive chemical changes of beef burgers treated with different antioxidants, namely, (a) a control without antioxidants, (b) 0.5 g/kg sodium erythorbate (ERY), (c) 2.5 g/kg encapsulated elderberry extract (EE 2.5), and (d) 5 g/kg encapsulated elderberry extract (EE 5), each one packaged under modified atmosphere (80% O2 and 20% CO2) for 13 days storage at 2 ± 1 °C, were deeply evaluated. Overall, EEs showed a wide array of antioxidant compounds, namely polyphenols like anthocyanins, flavonols, and phenolic acids. Multivariate statistics provided marked chemical differences between burgers manufactured with EEs and synthetic antioxidants (ERY) during 13-days storage in terms of both metabolomic profiles and typical lipid/protein oxidation markers (such as malondialdehyde and total carbonyls). Most of the differences could be attributed to some discriminant compounds, namely glutathione, 4-hydroxy-2-nonenal, hydroxy/peroxy-derivatives of fatty acids, carbonyl compounds (such as 5-nonen-2-one and 1,5-octadien-3-one), and cholesterol. Interestingly, significant correlations (p < 0.01) were observed between malondialdehyde, total carbonyls, and these discriminant metabolites. The combination of spectrophotometric approaches and a high-throughput untargeted metabolomics analysis outlined a strong modulation of both lipid and protein oxidations, likely promoted by the encapsulated meat extender (elderberry), thus confirming its ability to delay oxidative phenomena during the shelf-life of beef burgers.

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 the expression of the enzyme PON2 was observed in cancer cells. In conclusion, our data demonstrate that normal and cancer cells are differentially sensitive to CSE, which has different effects on PON2 gene expression as well. The overexpression of PON2 in T24 cells treated with CSE could represent a mechanism by which tumor cells protect themselves from the apoptotic process induced by glucosinolates and polyphenols.

Polyamines (PAs) are key signaling molecules involved in plant growth and stress acclimation processes. This work investigated the effect of spermidine, spermine, and putrescine (alone and in a mixture) in tomato plants using a combined metabolomics and lipidomics approach. The experiments were carried out under non-stress and 100 mM NaCl salinity conditions. Shoot and root biomass, as well as SPAD values, were increased by the application of exogenous PAs but with differences across treatments. Similarly, root length density (F: 34, p < 0.001), average root diameter (F: 14, p < 0.001), and very fine roots (0.0-0.5 mm) increased in PA-treated plants, compared to control. Metabolomics and lipidomics indicated that, despite being salinity the hierarchically prevalent factor, the different PA treatments imposed distinct remodeling at the molecular level. Plants treated with putrescine showed the broader modulation of metabolite profile, whereas spermidine and spermine induced a comparatively milder effect. The pathway analysis from differential metabolites indicated a broad and multi-level intricate modulation of several signaling molecules together with stress-related compounds like flavonoids and alkaloids. Concerning signaling processes, the complex crosstalk between phytohormones (mainly abscisic acid, cytokinins, the ethylene precursor, and jasmonates), and the membrane lipids signaling cascade (in particular, sphingolipids as well as ceramides and other glycerophospholipids), was involved in such complex response of tomato to PAs. Interestingly, PA-specific processes could be observed, with peculiar responses under either control or salinity conditions., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Ethnopharmacological relevance: Salvia glutinosa , also known as the glutinous sage, has been used in Romanian folk medicine in the treatment of inflammation, injuries, and mild infections. However, there is no direct scientific evidence to demonstrate these activities. Aim of the Study: The present research was based on evaluating antioxidant, antiproliferative, and α-glucosidase inhibitory activity of S. glutinosa extracts, as well as the in vivo anti-inflammatory activity. Materials and Methods: Infusions and 70% ( v:v ) ethanol solution extracts of S. glutinosa stems and leaves, collected from two different locations in Romania, were prepared. Ten phenolic compounds were identified and quantified using the LC-DAD-ESI/MS n method, and total phenolic and flavonoid content, as well as in vitro antioxidant (DPPH, ABTS, and FRAP assays), antiproliferative, anti-inflammatory and alpha-glucosidase inhibitory activities were determined. A rat model of induced inflammation with turpentine oil was used for the examination of in vivo effects of the extracts, using diclofenac as an anti-inflammatory control. Results: The highest inhibitory α-glucosidase activity was determined to be IC 50 = 0.546 mg/ml for the hydroalcoholic extract made with plant material collected on the road to Sighișoara. The highest cytotoxic activity against HepG2 cell line was determined to be GI 50 = 131.68 ± 5.03 μg/ml, for the hydroalcoholic extract made with plant material from Sighișoara. In vivo administration of extract (200 mg lyophilized powder/ml) showed a significant reduction of NO production. Conclusion: Our findings indicate that S. glutinosa extracts exhibit antioxidant, α-glucosidase inhibitory activity, as well as a modest cytotoxic effect on HepG2 cell line. By in vivo administration, the extracts show anti-inflammatory and antioxidant activity, which correlates with the traditional use of the species. The environmental conditions seemed to induce important changes in the chemical composition and the bioactivity of the herbal preparations derived from S. glutinosa., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Nicolescu, Babotă, Ilea, Dias, Calhelha, Gavrilaș, Rocchetti, Crișan, Mocan, Barros and Pârvu.)

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., Competing Interests: PB was employed by company oloBion-OMICS LIFE LAB. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ganugi, Fiorini, Rocchetti, Bonini, Tabaglio and Lucini.)

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., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Two techniques, namely, optimized ultrasound-assisted extraction (UAE) and enzyme-assisted extraction (EAE), were used to promote the extraction of phenolic compounds from the pseudo-fruits of Rosa canina L. (RC). For UAE, an optimization process based on the design of experiment (DoE) principles was used for determining the dependence between three variables (i.e., time of extraction, ultrasound amplitude, and the material-to-water ratio) and the total phenolic content of the samples. For EAE, a 2:1:1 pectinase, cellulase, and hemicellulase enzymatic blend was used as pre-treatment for optimized UAE, inducing a higher total phenolic content. The untargeted phenolic profiling approach revealed a great abundance of lower molecular weight phenolics (1.64 mg Eq./g) in UAE-RC extracts, whilst gallic acid (belonging to hydroxybenzoic acid derivatives) was the most abundant individual compound of both extracts. The unsupervised multivariate statistics clearly discriminated the impact of enzymatic pre-treatment on the phenolic profile of RC pseudo-fruits. Finally, Pearson’s correlation coefficients showed that anthocyanins, phenolic acids, and tyrosol derivatives were those compounds mostly correlated to the in vitro antioxidant potential of the extracts, whilst negative and significant (p < 0.05) correlation coefficients were recorded when considering the enzymatic inhibition activities. The highest enzyme-inhibitory activity has been identified against α-glucosidase, which indicates an antidiabetic effect.

Cross-adaptation phenomena in bacterial populations, induced by sublethal doses of antibacterial solutions, are a major problem in the field of food safety. In this regard, essential oils and their major compounds appear as an effective alternative to common sanitizers in food industry environments. The present study aimed to evaluate the untargeted metabolomics perturbations of Salmonella enterica serovar Enteritidis that has been previously exposed to the sublethal doses of the major components of essential oils: cinnamaldehyde, citral, and linalool (CIN, CIT, and LIN, respectively). Cinnamaldehyde appeared to be the most efficient compound in the assays evaluating the inhibitory effects [0.06% (v/v) as MBC]. Also, preliminary tests exhibited a phenotype of adaptation in planktonic and sessile cells of S . Enteritidis when exposed to sublethal doses of linalool, resulting in tolerance to previously lethal concentrations of citral. A metabolomics approach on S. Enteritidis provided an important insight into the phenomenon of cross-adaptation induced by sublethal doses of major compounds of some essential oils. In addition, according to the results obtained, when single molecules were used, many pathways may be involved in bacterial tolerance, which could be different from the findings revealed in previous studies regarding the use of phytocomplex of essential oils. Orthogonal projection to latent structures (OPLS) proved to be an interesting predictive model to demonstrate the adaptation events in pathogenic bacteria because of the global engagement to prevent and control foodborne outbreaks., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Pamplona Pagnossa, Rocchetti, Bezerra, Batiha, El-Masry, Mahmoud, Alsayegh, Mashraqi, Cocconcelli, Santos, Lucini and Hilsdorf Piccoli.)

The health benefits of extra virgin olive oil (EVOO) are far proven. However, considering that this oil is consumed also cooked, this work aimed to evaluate the impact of different cooking techniques on human colorectal adenocarcinoma cells (Caco-2) exposed to in vitro digested EVOO. In particular, the effect of different cooking methods, namely sauteing, deep-frying, and Roner®, was assayed and compared to a raw EVOO sample. The Caco-2 cell lysates were analyzed through an untargeted lipidomics approach, and multivariate statistics were used to identify the marker compounds of the differences in cells' lipidomic signatures. Despite representing the cooking at the lowest temperature (but longer time), cells exposed to Roner® cooked EVOO presented the most distinguished lipidomic profile. The markers of differences in Caco-2 could be related to oxidative stress-related compounds such as oxidized glutathione, diketogulonic acid, ceramides, and diglycerides. Taken together, our findings indicate that the differences in EVOO composition determined by cooking could impose significant lipidomic perturbation on the human intestinal cells., (Copyright © 2022. Published by Elsevier Ltd.)

In this work, the impact of terroir, cultivar, seasonality, and farming systems on functional traits of tomato was hierarchically investigated. Untargeted metabolomics, antioxidant capacity, colorimetric assays, and enzyme inhibition were determined. The total phenolic and carotenoid contents significantly varied between growing years, whereas an interaction between the farming system and growing year ( p < 0.01) was observed for total phenolics, carotenoids, and flavonoids, and for acetylcholinesterase inhibition. Hierarchical clustering showed that geographical origin and growing year were the major contributors to the differences in phytochemical profiles. Nonetheless, supervised modeling allowed highlighting the effect of the farming system. Several antioxidants (L-ascorbic acid, α-tocopherol, and 7,3',4'-trihydroxyflavone) decreased, whereas the alkaloid emetine and phytoalexin phenolics increased under organic farming. Taken together, our findings indicate that cultivar and pedo-climatic conditions are the main determinants for the functional quality of tomato, whereas the farming system plays a detectable but hierarchically lower., Competing Interests: PB was employed by company OloBion–OMICS LIFE LAB. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Rocchetti, Senizza, Zengin, Bonini, Bontempo, Camin, Trevisan and Lucini.)

In this work, the Longissimus thoracis pig skeletal muscle was used as a model to investigate the impact of two different diets, supplemented with n-3 polyunsaturated fatty acids from extruded linseed (L) and polyphenols from grape skin and oregano extracts (L+P), on the lipidomic profile of meat. A standard diet for growing-finishing pigs (CTRL) was used as a control. Changes in lipids profile were investigated through an untargeted lipidomics and transcriptomics combined investigation. The lipidomics identified 1507 compounds, with 195 compounds fitting with the MS/MS spectra of LipidBlast database. When compared with the CTRL group, the L+P diet significantly increased 15 glycerophospholipids and 8 sphingolipids, while the L diet determined a marked up-accumulation of glycerolipids. According to the correlations outlined between discriminant lipids and genes, the L diet may act preventing adipogenesis and the related inflammation processes, while the L+P diet promoted the expression of genes involved in lipids' biosynthesis and adipogenic extracellular matrix formation and functioning., Competing Interests: The authors have declared that no competing interests exist.

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.

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., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Sizeable scientific evidence indicates the health benefits related to phenolic compounds and dietary fiber. Various phenolic compounds-rich foods or ingredients are also rich in dietary fiber, and these two health components may interrelate via noncovalent (reversible) and covalent (mostly irreversible) interactions. Notwithstanding, these interactions are responsible for the carrier effect ascribed to fiber toward the digestive system and can modulate the bioaccessibility of phenolics, thus shaping health-promoting effects in vivo. On this basis, the present review focuses on the nature, occurrence, and implications of the interactions between phenolics and food components. Covalent and noncovalent interactions are presented, their occurrence discussed, and the effect of food processing introduced. Once reaching the large intestine, fiber-bound phenolics undergo an intense transformation by the microbial community therein, encompassing reactions such as deglycosylation, dehydroxylation, α- and β-oxidation, dehydrogenation, demethylation, decarboxylation, C-ring fission, and cleavage to lower molecular weight phenolics. Comparatively less information is still available on the consequences on gut microbiota. So far, the very most of the information on the ability of bound phenolics to modulate gut microbiota relates to in vitro models and single strains in culture medium. Despite offering promising information, such models provide limited information about the effect on gut microbes, and future research is deemed in this field., (© 2022 The Authors. Comprehensive Reviews in Food Science and Food Safety published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

In this study, the aerial parts and bulbs of nine Allium species were investigated for their functional phytochemical profile, in vitro antioxidant activities, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, α-glucosidase, and tyrosinase inhibitory properties. Phenolics, alkaloids, glucosinolates and other sulfur-containing compounds were distinctively profiled in the different species. Maceration in methanol allowed recovering the highest cumulative phenolic content in A. scabrifolium (42.31 mg/g), followed by A. goekyigiti (33.15 mg/g) and A. atroviolaceum (28.35 mg/g). The aerial parts of all Allium species showed high in vitro antioxidant activity whereas methanolic extract of A. cappadocicum bulb showed the highest inhibition against AChE (2.44 mg galantamine equivalent/g) and the water extracts of A. isauricum aerial part were the best BChE inhibitors (4.31 mg galantamine equivalent/g). Bulbs were the richer source of oligosaccharides, and in vitro digestion determined an increase of oligosaccharides bioaccessibility. A promising nutraceutical potential could be highlighted in our understudied Allium species., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: A high mortality rate is associated with about 80% of all infections worldwide, mainly due to antimicrobial resistance. Various antimicrobial and cytotoxic activities have been proposed for Meliaceae species. This study aimed to evaluate the in vitro anti-virulence and cytotoxic effect of the leaf extracts of Aphanamixis polystachya , Toona ciliata and Melia azedarach against five MRSA strains and on three cancer cell lines, followed by biological correlation to their encompassed phytoconstituents., Material and Methods: We explored three plants of this family against a panel of Methicillin-resistant Staphylococcus aureus (MRSA) strains and several cancer cell lines to select the most promising candidates for further in vivo and preclinical studies. The phytochemical composition was evaluated by UHPLC-QTOF-MS untargeted profiling. Cell viability was assessed by SRB assay. Minimum Inhibitory Concentration was carried out by using the agar micro-dilution technique. Inhibition of biofilm formation and preformed biofilm disruption were assessed spectrophotomertically, according to the Sultan and Nabil method (2019)., Results: A total of 279 compounds were putatively annotated to include different phytochemical classes, such as flavonoids (108), limonoids/terpenoids (59), phenolic acids (49) and lower-molecular-weight phenolics (39). A. polystachya extract showed the most potent cytotoxic activity against Huh-7, DU-145 and MCF-7 cell lines (IC 50 = 3, 3.5 and 13.4 µg mL -1 , respectively), followed by M. azedarach , with no effect recorded for T. ciliata extract. Furthermore, both A. polystachya and M. azedarach extracts showed promising anti-virulence and antimicrobial activities, with A. polystachya being particularly active against MRSA. These two latter extracts could inhibit and disrupt the biofilm, formed by MRSA, at sub-lethal concentrations. Interestingly, the extracts inhibited hemolysin-α enzyme, thus protecting rabbit RBCs from lysis. A. polystachya extract reduced the pigmentation and catalase enzyme activity of tested pigmented strains better than M. azedarach at both tested sub-MICs. Consequently, susceptibility of the extract-treated cells to oxidant killing by 200 mM H 2 O 2 increased, leading to faster killing of the cells within 120 min as compared to the extract-non-treated cells, likely due to the lower antioxidant-scavenging activity of cells exhibiting less staphyloxanthin production., Conclusion: These findings suggested that both A. polystachya and M. azedarach natural extracts are rich in bioactive compounds, mainly limonoids, phenolics and oxygenated triterpenoids, which can combat MRSA biofilm infections and could be considered as promising sources of therapeutic cytotoxic, antibiofilm and anti-virulence agents.

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., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

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., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This work investigated the microbiological quality and chemical profiles of two different dairy creams obtained by centrifugation vs. natural creaming separation systems. To this aim, an untargeted metabolomics approach based on UHPLC-QTOF mass spectrometry was used in combination with multivariate statistical tools to find potential marker compounds of the two different types of two dairy creams. Thereafter, we evaluated the chemical, microbiological and sensorial changes of a ricotta cheese made with a 30% milk cream (i.e., made by combining dairy creams from centrifugation and natural creaming separation) during its shelf-life period (12 days). Overall, microbiological analysis revealed no significant differences between the two types of dairy creams. On the contrary, the trend observed in the growth of degradative bacteria in ricotta during shelf-life was significant. Metabolomics revealed that triacylglycerols and phospholipids showed significant strong down-accumulation trends when comparing samples from the centrifugation and natural creaming separation methods. Additionally, 2,3-Pentanedione was among the best discriminant compounds characterising the shelf-life period of ricotta cheese (VIP score = 1.02), mainly related to sensorial descriptors, such as buttery and cheesy. Multivariate statistics showed a clear impact of the shelf-life period on the ricotta cheese, revealing 139 potential marker compounds (mainly included in amino acids and lipids). Therefore, the approach used showed the potential of a combined metabolomic, microbiological and sensory approach to discriminate ricotta cheese during the shelf-life period.

Seedlings of Rumex acetosa L. (sorrel) were grown in floating system and two consecutive cuts took place: 15 (C1) and 30 (C2) days after sowing. An untargeted metabolomics approach was utilised to fingerprint phenolics and other health-related compounds in sorrel leaves, as well as to unveil differences between the two cuts. The untargeted approach allowed to putatively identify 458 metabolites considering both the cuts. Three new terpenoids and two new phenylpropanoid glycosides (dihydrosyringin and dihydroconiferin) with antifungal and anti-inflammatory activity were annotated. Overall, leaves from C2 had lower level of secondary metabolites (44 were down-accumulated), especially sesquiterpenes and stilbenes. Conversely, anthocyanins showed a relevant increase in C2 than in C1 leaves. The dataset suggests that sorrel leaves represent a good source of nutraceutical compounds and unveils the pivotal effect of pre-harvest factor in secondary metabolite profile.