Search

Food proteins are an important source of bioactive peptides, and the angiotensin I-converting enzyme (ACE) inhibitors are worthy of attention for their possible beneficial effects in subjects with mild hypertension. However, the chemical basis underpinning their activity is not well-understood, hampering the discovery of novel inhibitory sequences from the plethora of peptides encrypted in food proteins. This work combined computational and

In Mediterranean areas, agricultural systems have to adapt to an environment presenting few water resources with severe drought events, which are expected to increase in frequency and intensity due to global warming. Therefore, these regions are particularly exposed to climate change and need to implement solutions in order to maintain food production. Our objective is to assess the benefits of innovative cropping systems capable to face these constraints. Evolutionary population are mixtures of plants of the same species presenting a high degree of crop genetic diversity needing lower inputs while allowing higher buffering capacities to adapt environmental stress like water shortage. In this study, we investigated the influence that these plants can have on organic matter quality and dynamics and hypothesised that the abilities of the crop to face drought can participate at increasing soil carbon storage in soil. We conducted a field experiment with five different evolutionary populations of wheat, i.e. a bread wheat (Monnalisa), an einkorn wheat (Norberto) and 2 evolutionary populations (Furat-Li Rosi, Furat-Floriddia and BIO2), cultivated following four other plants species (Wheat, Pea, Cickpea or Clover) on two different sites in Italy presenting contrasted conditions (hot-summer Mediterranean climate in Roma and humid subtropical climate in Parma). Bulk soil and rhizosphere soils were sampled and C, N contents as well as organic matter functional groups using Mid Infrared Spectroscopy (MIRS) were assessed. We observed that the infrared signatures differed between our two sites and depending on the previous cropping species. Evolutionary population of wheat showed different signatures than durum and bread wheat. Our results demonstrate that evolutionary population in particular pedoclimatic conditions can influence the fate of soil organic matter.

In recent years, due to the significant increase in hypertension, peptides which are able to reduce blood pressure have gained special interest by scientific research and food industry. Several bioactive peptides with ascertained ACE-inhibitory activity have been found in Parmigiano Reggiano (PR) cheese and/or mixtures deriving from its digestion in vitro, and this may be predictive of its potential antihypertensive effect in vivo. This study investigated the long-term effect of feeding (PR) cheese on blood pressure (BP) of spontaneously hypertensive rats (SHRs). A total of 30 male SHRs, 13 weeks old, were subdivided into 6 groups balanced for body weight and BP, to receive daily dietary supplementation with: 0.1–0.2–0.4–0.6 g PR/rat, captopril, and water. Systolic and diastolic BP were recorded every two weeks, for 10 weeks. Blood samples were collected at the end of the trial. Dietary integration with PR led to a transitory reduction in rats’ pressure in the first 35 days of treatment and pressure decreased in a dose-dependent manner. In the second part of the study, the beneficial effect of PR antihypertensive peptides may have been masked and reduced by the increase in BP of rats linked to the rise in age of animals. No PR derived peptides were detected in rats’ serum. Highlights: Parmigiano Reggiano (PR) cheese led to a transitory reduction in rats’ pressure in the first 35 days of treatment. This effect was PR dose dependent. The highest amounts of PR tested did not increase both systolic and diastolic blood pressures of hypertensive rats.

In the last years, due to the increasing interest of consumers in natural products, market and industry choices were more directed towards the use of fragrances and flavors obtained from natural sources. Fermentation is a biological approach helpful to produce diverse aromatic compounds modifying precursors present in raw material, due to a wide array of extracellular enzymes which could be produced by microorganisms. Following this direction Lacticaseibacillus rhamnosus was employed as a starter for melon and orange by-products fermentation. Simple and vacuum distillation were applied for the recovery of the aromatic fraction and compared to identify the more promising in terms of aromatic profile composition. Detection and characterization of aromatic compounds were achieved using Head SpaceSolid Phase MicroExtraction Gas Chromatography–Mass Spectrometry (HS-SPME/GC–MS) approach. Fermentation led to interesting changes, increasing floral, herbal green and fruity notes. Overall, simple distillation allowed the greater recovery of aromatic compounds in terms of concentration for both substrates. This study highlights a possible strategy to produce complex distillates, composed of a mixture of aroma compounds, to be employed as additives in food, feed or perfumery industries as ingredients of natural origin.

This chapter aims to address an issue of ancient origins, but more and more topical in a globalized world in which consumers and stakeholders are increasingly aware: the authenticity of food. Foods are systems that can also be very complex, and verifying the correspondence between what is declared and the actual characteristics of the product is often a challenging issue. The complexity of the question we want to answer (is the food authentic?) means that the answer is equally articulated and makes use of many different analytical techniques. This chapter will consider the chemical analyses of foods aimed at guaranteeing their authenticity and will focus on frontier methods that have been developed in recent years to address the need to respond to ever-increasing guarantees of authenticity. Targeted and non-targeted approaches will be considered for verifying the authenticity of foods, through the study of different classes of constituents (proteins, metabolites, lipids, flavors). The numerous approaches available (proteomics, metabolomics, lipidomics) and the related analytical techniques (LC-MS, GC-MS, NMR) are first described from a more general point of view, after which their specific application for the purposes of authentication of food is addressed.

Hepatitis E Virus (HEV) follows waterborne or zoonotic/foodborne transmission. Genotype 3 HEV infections are worldwide spread, especially in swine populations, representing an emerging threat for human health, both for farm workers and pork meat consumers. Unfortunately, HEV in vitro culture and analysis are still difficult, resulting in a poor understanding of its biology and hampering the implementation of counteracting strategies. Indeed, HEV encodes for only one non-structural multifunctional and multidomain protein (ORF1), which might be a good candidate for anti-HEV drugging strategies. In this context, an in silico molecular modelling approach that consisted in homology modelling to derive the 3D model target, docking study to simulate the binding event, and molecular dynamics to check complex stability over time was used. This workflow succeeded to describe ORF1 RNA Helicase domain from a molecular standpoint allowing the identification of potential inhibitory compounds among natural plant-based flavagline-related molecules such as silvestrol, rocaglamide and derivatives thereof. In the context of scouting potential anti-viral compounds and relying on the outcomes presented, further dedicated investigations on silvestrol, rocaglamide and a promising oxidized derivative have been suggested. For the sake of data reproducibility, the 3D model of HEV RNA Helicase has been made publicly available.

The market value of vanilla is constantly growing, as it is the aroma most appreciated by consumers worldwide. The key component of the aroma of vanilla beans is vanillin, which can be directly extracted from the plant, produced by chemical synthesis, or by bioconversion of natural precursors. Due to the increasing consumers' demand for products labeled as “natural,” extraction from vanilla pods results in a more valuable aroma source. Once the extraction is completed, what remains are the exhausted beans that still contain small seeds and other compounds, including varying amounts of vanillin trapped in the cellular structures of the plant. The application of fermentation of exhausted vanilla beans is proposed here as a strategy to recover “natural” vanillin and other valuable aroma compounds as a result of the metabolic conversion by lactic acid bacteria (LAB). The aim of this study was to verify the fermentability of exhausted vanilla beans by-products for their valorization, allowing the recovery of high-value molecules or new applications in food products. Design of Experiment (DoE) was used to screen a library of LAB strains to identify the best condition of fermentation in response to varying cultivation conditions. A comparison between mono and co-culture of LAB was assessed. Moreover, sensory panel tests and the evaluation of the aromatic components by Solid Phase Micro Extraction-Gas Chromatography-Mass Spectrometry analysis were carried out to better understand the modification of the aroma profile after fermentation. Fermentation with LAB changed the volatile profile and sensory characteristics of the exhausted vanilla beans and represents a promising method for the valorization of these by-products.

Kokumi is an important taste perception whose chemical basis still needs clarifications and for which the development of high-throughput tools of analysis is desirable. The activation of Calcium-sensing receptor (CaSR) was described as the basis of kokumi perception allowing the use of molecular modelling to deepen its chemical rationale and related mechanisms. This study focused on γ-Glutamyl tripeptides, computationally providing mechanistic insights on their CaSR-activating properties and extended the comprehension of their structure-activity relationship. A library of 400 γ-Glutamyl tripeptides was also screened. γ-Glu-Pro-Ala and γ-Glu-Pro-Ser were identified for further dedicated investigations based on their promising CaSR-activating potential and their presence should be checked accordingly in food matrices to better profile the kokumi fingerprint. This work provided a meaningful tool for the top-down analysis of kokumi-active molecules that may support either the identification of kokumi molecules concealed in food or the rational design of kokumi-active molecules de novo.

LTFPGSAED (P7) is a multifunctional hypocholesterolemic and hypoglycemic lupin peptide. While assessing its angiotensin-converting enzyme (ACE) inhibitory activity, it was more effective in intestinal Caco-2 cells (IC50 of 13.7 μM) than in renal HK-2 cells (IC50 of 79.6 μM). This discrepancy was explained by the metabolic transformation mediated by intestinal peptidases, which produced two main detected peptides, TFPGSAED and LTFPG. Indeed LTFPG, dynamically generated by intestinal dipeptidyl peptidase IV as well as its parent peptide P7 were linearly absorbed by mature Caco-2 cells. An in silico study demonstrated that the metabolite was a better ligand of the ACE enzyme than P7. These results are in agreement with an in vivo study, previously performed by Aluko et al., which has shown that LTFPG is an effective hypotensive peptide. Our work highlights the dynamic nature of bioactive food peptides that may be modulated by the metabolic activity of intestinal cells.

Rice (Oryza sativa L.) bran is an important by‐product produced during the milling of brown rice, rich in several valuable compounds, such as lipids. Moreover, considering the valorisation potential of these material, the application of innovative and low‐impact techniques can improve the overall quality of by‐products for their future exploitation in the food manufacturing. In this study, the impact of air classification and micronisation (fine grinding) of rice bran on the lipid components was studied. These treatments allowed to obtain bran fractions, with different granulometry, from coarse to very fine. The total crude fat content was significantly higher in fine air‐classified rice bran fractions. Besides, polyunsaturated triacylglycerols (TAG) were the most abundant compounds and monoacylglycerols concentration increased from ~15% in rice bran to ~22% in fine fraction. Considering the relevant emulsifying properties of these compounds, this fraction could be used as functional ingredient for the quality improvement of cereal‐based products, without modifying the valuable fatty acid profile.

IAVPTGVA (Soy1) and LPYP are two soybean peptides, which display a multifunctional behavior, showing in vitro hypocholesterolemic and hypoglycemic activities. A preliminary screening of their structures using BIOPEP suggested that they might be potential angiotensin-converting enzyme (ACE) inhibitors. Therefore, a bottom-up-aided approach was developed in order to clarify the in vitro hypotensive activity. Soy1 and LPYP dropped the intestinal and renal ACE enzyme activity with IC50 values equal to 14.7 ± 0.28 and 5.0 ± 0.28 μM (Caco-2 cells), and 6.0 ± 0.35 and 6.8 ± 0.20 μM (HK-2 cells), respectively. In parallel, a molecular modeling study suggested their capability to act as competitive inhibitors of this enzyme. Finally, in order to increase both their stability and hypotensive properties, a suitable strategy for the harmless control of their release from a nanomaterial was developed through their encapsulation into the RADA16-assembling peptide.

Non-thermal processing of milk has been considered over the past decade as an alternative or adjunct to thermal processes. UV-C radiation in combination with turbulent flow of opaque liquids seems to be a promising non-thermal method for the reduction of bacterial populations in milk. Apart from confirming the efficacy of UV-C in destroying pathogens and spoilage bacteria, there is a need for assessment of the quality characteristics of the end-product and especially in added-value dairy products where bioactivity of constituents should be preserved during processing. Under this context, freeze-dried donkey milk powder processing by UV-C was studied and the effect on protein quality, digestibility and bioactive properties were assessed. Results show that UV-C treatment retains the protein's quality characteristics highly comparable to the not-treated milk (i.e. raw) rather than the pasteurized milk where some deterioration (i.e. lower bioactivities) was detected.

Reducing the salt content in food is an important nutritional strategy for decreasing the risk of diet-related diseases. This strategy is particularly effective when applied to highly appreciated food having good nutritional characteristics, if it does not impact either upon sensory or nutritional properties of the final product. This work aimed at evaluating if the reduction of salt content by decreasing the brine soaking time modifies fatty acid and protein bioaccessibility and bioactive peptide formation in a 30-month-ripened Parmigiano Reggiano cheese (PRC). Hence, conventional and hyposodic PRC underwent in vitro static gastrointestinal digestion, and fatty acid and protein bioaccessibility were assessed. The release of peptide sequences during digestion was followed by LC–HRMS, and bioactive peptides were identified using a bioinformatic approach. At the end of digestion, fatty acid and protein bioaccessibility were similar in conventional and hyposodic PRC, but most of the bioactive peptides, mainly the ACE-inhibitors, were present in higher concentrations in the low-salt cheese. Considering that the sensory profiles were already evaluated as remarkably similar in conventional and hyposodic PRC, our results confirmed that shortening brine soaking time represents a promising strategy to reduce salt content in PRC.

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.

A sensitive UHPLC-ESI-MS/MS method, coupled with a suitable extraction protocol, was developed and validated for the rapid detection of four tropane alkaloids (anisodamine, atropine, homatropine, scopolamine) in herbal teas and herbal extracts. The method offers good sensitivity, accuracy and precision. The protocol is suitable for routine determination of tropane alkaloids in dried plants and extracts sold as food supplements or phytotherapics, and was applied to 60 commercial samples. Anisodamine, atropine and/or scopolamine were found in 4 herbal teas samples (LOQ–69 μg/kg for atropine and 50 μg/kg for scopolamine) and in a Lepidium meyenii extract sold as food supplement (traces of anisodamine). The level of contamination do not to pose a direct risk for the consumers if compared to the acute reference dose indicated by EFSA, but the detection confirms the need for suitable methods and careful screenings in order to determine a more precise evaluation of dietary intake for tropane alkaloids.

Mycotoxins in food may threat public health at a global scale. However, for most of them, the current body of knowledge does not support a proper risk assessment and more data are needed to clarify their toxicity. In particular, the assessment of “personalized” action may succeed in understanding and counteracting the effects of many toxicants. Therefore, the assessment of “personalized” toxicology of mycotoxins might deserve attention to foster the understanding of their mechanisms of toxicity and to eventually improve the assessment of risk. This work dealt with the early warning analysis of possible differences in eliciting androgenic stimuli by alternariol, a widespread mycotoxin produce by Alternaria species, when mutations on the androgen receptor occur. It was applied a computational study based on docking simulations, pharmacophore modeling and molecular dynamics to assess the capability of alternariol to interact with the androgen receptor bearing the M749I substitution – which confers insensitivity to androgens stimulation. The results collected pointed to possible “protective” effects against alternariol suggesting: i) the likely existence of inter-individual responses to alternariol stimulation; ii) the meaningfulness of collecting data on “personalized” response to mycotoxins toward a more precise paradigm addressing the risk assessment at the individual level.

Mycotoxins are one of the most important contaminants in cereal grains. Besides parent forms, the presence and identification of structurally modified mycotoxins is nowadays recognized as a challenging food safety-related issue and contribute to increase the human and animal exposure. The aim of this study was to follow the distribution of Fusarium toxins and their main modified forms in the pearled fractions of several grain species (i.e. tritordeum, durum and bread wheat, and barley), using high-resolution mass spectrometry technique (HR-MS). A significant decreasing trend in mycotoxins concentration was observed from the outer layer to the inner kernel, along the sequential pearling process. Among modified forms, deoxynivalenol (DON) -oligoglucosides were described for the first time in naturally infected grains, while zearalenone (ZEN) -sulphate was the only ZEN-related form detected in pearling fractions. HR-MS could be confirmed as useful technique to study and characterize modified forms of mycotoxins.

Pistachio hull is a substantial source of natural polyphenols, but a substantial volume is being wasted annually. Aqueous pistachio green hull extract (PGHE) was subjected to two-stage membrane process in order to separate a polyphenol rich fraction. The membrane conditions of each stage were investigated. Total phenolic content and antioxidant activity were determined by Folin–Ciocalteu and DPPH · assay; also membrane fouling was monitored. The use of the 1 kDa cellulose membrane accompanied by 4 bar pressure and 250 rpm stirring speed was observed to be successful in the separation of a fraction with the highest amount of phenolic compound and antioxidant activity, in the retentate part. UHPLC/MSn characterization of PGHE polyphenols enabled us to identify 34 compounds, including the most abundant galloylshikimic acids, gallic acid, theogallin, galloyl-O-hexoside, quercetin-O-hexoside and pyrogallol.

The aim of the present work was to evaluate the effect of sample particle size on fumonisins recovery during the extraction step. Four maize samples were ground and the resulted flours were separated in six different fractions according to their particle size (1000-250 μm). Fumonisin B1 and B2 were quantified on each fraction, as well as on the unfractionated sample by HPLC-MS/MS. Furthermore, the proximate analysis was carried out to exclude the influence of macro-constituent on the mycotoxins distribution. Although the six maize fractions were found to be characterized by the same macro-constituent composition, a significant increase (p

Bioactive peptides are short peptides (3–20 amino acid residues in length) endowed of specific biological activities. The identification and characterization of bioactive peptides of food origin are crucial to better understand the physiological consequences of food, as well as to design novel foods, ingredients, supplements, and diets to counteract mild metabolic disorders. For this reason, the identification of bioactive peptides is also relevant from a pharmaceutical standpoint. Nevertheless, the systematic identification of bioactive sequences of food origin is still challenging and relies mainly on the so defined “bottom-up” approaches, which rarely results in the total identification of most active sequences. Conversely, “top-down” approaches aim at identifying bioactive sequences with certain features and may be more suitable for the precise identification of very potent bioactive peptides. In this context, this work presents a top-down, computer-assisted and hypothesis-driven identification of potent angiotensin I converting enzyme inhibitory tripeptides, as a proof of principle. A virtual library of 6840 tripeptides was screened in silico to identify potential highly potent inhibitory peptides. Then, computational results were confirmed experimentally and a very potent novel sequence, LMP was identified. LMP showed an IC50 of 15.8 and 6.8 µM in cell-free and cell-based assays, respectively. In addition, a bioinformatics approach was used to search potential food sources of LMP. Yolk proteins were identified as a possible relevant source to analyze in further experiments. Overall, the method presented may represent a powerful and versatile framework for a systematic, high-throughput and top-down identification of bioactive peptides.

Okara is the main soybean by-product resulting from the processing of soy milk and tofu. Despite being a product with a lot of potential and rich in many bioactive compounds such as polyphenols, it presents an unpleasant, rancid aroma. For this reason its use in the food industry is limited. In this study, we have reported the integral use of okara in a solid state fermentation process, conducted with wild strains of lactic acid bacteria, to evaluate the effect of bacterial metabolism on the volatile and polyphenolic profiles. Strains belonging to Lactobacillus acidophilus, Lacticaseibacillus rhamnosus and Pediococcus acidilactici species were used in monoculture and, for the first time, in co-culture. The results showed an improvement in the aromatic fraction showing a decrease in hexanal, responsible for off-flavour, and an increase in ketones with fruity and buttery notes in fermented okara. Polyphenols were also affected, and, in particular, a bioconversion of glucoside isoflavones to the aglycone forms was highlighted in all fermented substrates. In addition, the appearance of both phenyllactic and p-hydroxyphenyllactic acids as well as the increase in indole-3-lactic acid was observed for the first time upon okara fermentation. Overall, the co-culture appears to be the most promising for biovalorization of okara, thereby opening the possibility of its use in the development of functional ingredients.

Brine soaking is one of the most important steps in the production of Parmigiano Reggiano cheese, since it determines the amount of salt in the final product. Reduction in salt in Parmigiano Reggiano cheese might be important for improving its nutritional profile, but it could affect the manufacturing processes by altering proteolysis and consequently the product quality. In this study, for the first time, salt reduction was explored at the industrial level on real cheese samples manufactured in a local dairy. In particular, 20 wheels were produced with conventional (18 days, 10 wheels) and shorter (12 days, 10 wheels) brining steps. In every group, wheels were studied at two different ripening times, 15 and 30 months. A shorter brining time resulted in an average 12% decrease in salt content. A full characterization of free amino acids and peptides was performed by LC-MS on all samples. Free amino acids and peptides, as expected, increased with ripening, due to proteolysis, with samples having low salt content showing a slightly faster increase when compared to standard ones, hinting to a slightly accelerated proteolytic process. Nonetheless, low-salt and conventional cheeses shared similar sensory profiles at both ripening times.

This article belongs to the Special Issue Recovery of High Value-Added Compounds from Food By-Product., The aim of this study is to review the innovative techniques based on bioprocessing, thermal or physical treatments which have been proposed during the last few decades to convert rice bran into a valuable food ingredient. Rice bran (Oryza sativa) is the main by-product of rice grain processing. It is produced in large quantities worldwide and it contains a high amount of valuable nutrients and bioactive compounds with significant health-related properties. Despite that, its application in food industry is still scarce because of its sensitivity to oxidation processes, instability and poor technological suitability. Furthermore, the health-related effects of pretreated rice bran are also presented in this review, considering the up-to-date literature focused on both in vivo and in vitro studies. Moreover, in relation to this aspect, a brief description of rice bran arabinoxylans is provided. Finally, the application of rice bran in the food industry and the main technology aspects are concisely summarized., M.S. received a PhD grant by Regione Emilia-Romagna, under the scheme POR-FSE/2016. This research was partially supported by “Nuevos conocimientos para la sostenibilidad del sector cafetero” funded by Consejo Superior de Investigaciones Científicas (CSIC) (201970E117) and “Novel coffee by-product beverages for an optimal health of the brain-gut axis (COFFEE4BGA)” funded by the Ministerio de Ciencia e Innovación (PID2019-111510RB-I00).

Scope Urolithin A and B are gut metabolites of ellagic acid and ellagitannins associated with many beneficial effects. Evidence in vitro pointed to their potential as estrogenic modulators. However, both molecular mechanisms and biological targets involved in such activity are still poorly characterized, preventing a comprehensive understanding of their bioactivity in living organisms. This study aimed at rationally identifying novel biological targets underlying the estrogenic-modulatory activity of urolithins. Methods and results The work relies on an in silico/in vitro target fishing study coupling molecular modeling with biochemical and cell-based assays. Estrogen sulfotransferase and 17β-hydroxysteroid dehydrogenase are identified as potentially subject to inhibition by the investigated urolithins. The inhibition of the latter undergoes experimental confirmation either in a cell-free or cell-based assay, validating computational outcomes. Conclusions The work describes target fishing as an effective tool to identify unexpected targets of food bioactives detailing the interaction at a molecular level. Specifically, it described, for the first time, 17β-hydroxysteroid dehydrogenase as a target of urolithins and highlighted the need of further investigations to widen the understanding of urolithins as estrogen modulators in living organisms.

Ochratoxin A (OTA), a mycotoxin that is of utmost concern in food and feed safety, is produced by fungal species that mainly belong to the Aspergillus and Penicillium genera. The development of mitigation strategies to reduce OTA content along the supply chains is key to ensuring safer production of food and feed. Enzyme-based strategies are among the most promising methods due to their specificity, efficacy, and multi-situ applicability. In particular, some enzymes are already known for hydrolyzing OTA into ochratoxin alpha (OT&alpha, ) and phenylalanine (Phe), eventually resulting in detoxification action. Therefore, the discovery of novel OTA hydrolyzing enzymes, along with the advancement of an innovative approach for their identification, could provide a broader basis to develop more effective mitigating strategies in the future. In the present study, a hybrid in silico/in vitro workflow coupling virtual screening with enzymatic assays was applied in order to identify novel OTA hydrolyzing enzymes. Among the various hits, porcine carboxypeptidase B was identified for the first time as an effective OTA hydrolyzing enzyme. The successful experimental endorsement of findings of the workflow confirms that the presented strategy is suitable for identifying novel OTA hydrolyzing enzymes, and it might be relevant for the discovery of other mycotoxin- mitigating enzymes.

Plant biodiversity preservation is one of the most important priorities of today’s agriculture. Wheat (Triticum spp. L.) is widely cultivated worldwide, mostly under a conventional and monovarietal farming method, leading to progressive biodiversity erosion. On the contrary, the evolutionary population (EP) cultivation technique is characterized by mixing and sowing together as many wheat genotypes as possible to allow the crop to genetically adapt over the years in relation to specific pedoclimatic conditions. The objective of this study was to assess the nutritional, chemical and sensory qualities of three different breads obtained using different organic EP flours, produced following a traditional sourdough process and compared to a commercial wheat cultivar bread. Technological parameters, B-complex vitamins, microelements, dietary fibre and phenolic acids were determined in raw materials and final products. Flours obtained by EPs showed similar characteristics to the commercial wheat cultivar flour. However, significant differences on grain technological quality were found. The breads were comparable with respect to chemical and nutritional qualities. Overall, the sensory panellists rated the tasted breads positively assigning the highest score to those produced with EPs flours (6.75–7.02) as compared to commercial wheat cultivar-produced bread (cv. Bologna, 6.36).

This study aimed to evaluate the impact of breads made with two different wheat evolutionary populations (EPs), in comparison with a modern variety, on postprandial blood glucose and insulin responses. A randomized controlled crossover postprandial study involving twelve healthy subjects was conducted. Seven non-commercial breads produced with flours from two different bread wheat (T. aestivum L.) EPs (Bio2, ICARDA) and a modern bread wheat variety (Bologna) considered as control, and with two different bread-making processes (S. cerevisiae and sourdough), were specifically formulated for the study. Postprandial incremental curves, Incremental Area Under the Curve (IAUC), maximum postprandial peaks for blood glucose and plasma insulin over 2 hours after administration of isoglucidic portions of breads (50 g of available carbohydrates) were evaluated. The comparison of incremental curves, IAUC, and maximum postprandial peaks after consumption of breads formulated with EPs and control breads showed no differences among samples. Neither the flour nor the leavening technic used for the baking were effective in inducing a different postprandial response compared to the Bologna variety. EPs, being characterized by higher degree of crop genetic diversity, may have a relevant agronomic role to guarantee good and stable yields and quality under low input management in a changing climate, however, future studies are needed to better investigate their potential positive effect on human health.

Proteolysis is the most important event occurring during maturation of dry-cured hams: it strongly influences the flavour and the texture of the aged ham by the accumulation of peptides and free amino acids released by protein hydrolysis. Apart from compounds of proteolytic origin, it has been demonstrated that also non-proteolytic amino acyl derivatives (γ-glutamyl amino acids, pyroglutamyl-amino acids and lactoyl-amino acids) may accumulate during ripening of cheese, and they can be also found in fermented soy sauce, where they contribute to the umami taste of the products. Using a semi-quantitative analysis, in this paper we report the occurrence of significant amounts of γ-glutamyl amino acids and, for the first time, pyroglutamyl-amino acids and lactoyl-amino acids, in aged ham. The amino acid counterparts were mainly found to be hydrophobic amino acids. The amount of these compounds was found to increase with time, because they are not degraded by proteolytic activity. They were also found to be stable to simulated gastrointestinal digestion. Angiotensin Converting Enzyme inhibitory activity was also tested, but they were not found to be characterized by significant ACE-inhibitory activity.

Due to favourable climate condition, Italy is a prominent producer of different wheat varieties. Several wheat baked goods are produced, but the most typical Italian foods, like pasta, pizza and bread, are made of durum and common wheat flour. Because of the great importance of wheat in the Italian food market, authenticity represents an essential quality parameter not only for the producers and regulatory bodies but also for consumers. The aim of our study was to test the effectiveness of an unconventional non-targeted method for the discrimination of Triticum species using direct analysis real time-high-resolution mass spectrometry (DART-HRMS). For this purpose, 60 wheat samples including durum, common and hulled wheat varieties were collected over two consecutive harvest years. Chemometric evaluation revealed an optimal sample clustering according to the wheat species and the presence of 18 significant markers able to discriminate the groups. The discrimination power obtained is promising since the use of DART-HRMS can significantly reduce the analysis time compared to chromatographic techniques. A plausible future commercial and industrial scenario could see the application of this analytical approach especially to evaluate the risk of substitution of higher value wheat species with lower value flours.

Grape pomace, the major byproduct of the wine and juice industry, is a relevant source of bioactive phenolic compounds. However, polyphenol bioavailability in humans is not well understood, and the inter-individual variability in the production of phenolic metabolites has not been comprehensively assessed to date. The pharmacokinetic and excretive profiles of phenolic metabolites after the acute administration of a drink made from red grape pomace was here investigated in ten volunteers. A total of 35 and 28 phenolic metabolites were quantified in urine and plasma, respectively. The main circulating metabolites included phenyl-gamma-valerolactones, hydroxybenzoic acids, simple phenols, hydroxyphenylpropionic acids, hydroxycinnamates, and (epi)catechin phase II conjugates. A high inter-individual variability was shown both in urine and plasma samples, and different patterns of circulating metabolites were unravelled by applying unsupervised multivariate analysis. Besides the huge variability in the production of microbial metabolites of colonic origin, an important variability was observed due to phase II conjugates. These results are of interest to further understand the potential health benefits of phenolic metabolites on individual basis.

A model was set up to elucidate the uptake, translocation, and metabolic fate of zearalenone (ZEN) in durum wheat. After treatment with ZEN, roots and shoots were profiled with LC-HRMS. A comprehensive description of in planta ZEN biotransformation and a biotechnological evaluation of the model were obtained. Up to 200 μg ZEN were removed by each plantlet after 14 days. Most ZEN and its masked forms were retained in roots, while minimal amounts were detected in leaves. Sixty-two chromatographic peaks were obtained, resulting in 7 putative phase I and 18 putative phase II metabolites. ZEN16Glc and ZEN14Glc were most abundant in roots, sulfo-conjugates and zearalenol derivatives were unable to gain systemic distribution, while distinct isomers of malonyl conjugates were found in leaves and roots. This study underlines the potential ZEN occurrence in plants without an ongoing Fusarium infection. Micropropagation may represent a tool to investigate the interplay between mycotoxins and wheat.

In the present work the possibility of using an untargeted metabolomic strategy to discriminate between common and durum wheat lipidome for an authenticity purpose was explored. A first study was conducted by analyzing 52 samples from two durum and common wheat varieties. Afterwards, an extended and independent sample set (173 samples and five varieties) was used as a confirmatory study to verify the stability and consistency of the models obtained. Putatively identified markers were evaluated applying ROC curves resulting in individual marker AUC >90% both in preliminary and confirmatory study. In addition, digalactosyl diglyceride (DGDG) 36:4 was shown to be an effective marker differentiating between authentic durum wheat and its adulterated admixture down to 3% adulteration level, which is the maximum contamination level allowed by Italian legislation. The results demonstrated that untargeted lipidomics, in conjunction with chemometric tools has a significant potential for screening and detection of wheat fraud.

Tetrodotoxin (TTX) is a potent neurotoxin naturally found in terrestrial and marine animals targeting the voltage-gated sodium channels. Historically, TTX has raised food safety concerns mainly in the Asian countries due to the consumption of the typical pufferfish-derived delicacy fugu. However, the diffusion of TTX is getting wider today, reasonably threating in a close future a broader number of consumers than before. The understanding of TTX group toxicity is still incomplete as most of the analogues and metabolites found together with TTX are largely understudied. This prevents the establishment of a solid background for risk assessment and additional data have been claimed to timely foster the assessment of TTX toward a group-based approach. However, the high costs in sourcing TTX analogues make practically unfeasible the wide-scale assessment using experimental trials. The toxicological assessment in silico may succeed in extending data on compounds poorly affordable, hierarchizing compound to focus experiments and supporting the hazard identification. Therefore, the present work investigated the toxicodynamic of TTX, analogues and metabolites by using a molecular modeling approach. In the framework of the hazard identification, the model analyzed TTX analogues never tested before assessing qualitatively their potential toxicity in comparison to TTX. While the analogues from TTX bearing species appeared to be less toxic than TTX, some human metabolites showed a better interaction with the toxin binding site. Such results suggest that human metabolism may partially fail in preventing the interaction with the biological target. Therefore, the identification and assessment of human metabolites should be done to support the decision making process from a more informed perspective.

The mycotoxin zearalenone may contaminate food and feed worldwide upon infections by Fusarium spp. of plants and raw materials intended for human and animal consumption. Currently, contamination by zearalenone and congeners pose concern for health due to xenoestrogenic effects. However, while zearalenone and the main reduced metabolites are well-known xenoestrogens, some plant metabolites that may enter the food chain have been observed aside. Among them, zearalenone-14-glucoside may be abundant in the edible parts of infected plants, thereby entering significantly the human diet and animal feeding. On the basis of previous works, the lack of xenoestrogenicity for this compound per se was taken for granted, while neglecting the direct proof of estrogenic activity and considering the hydrolysis as a possible source of estrogenically active metabolites. The present work investigated the xenoestrogenicity of zearalenone-14-glucoside, in comparison to zearalenone, deepening the underlying molecular mechanisms through an integrated in vitro/in silico approach. On the basis of our results, zearalenone-14-glucoside effectively stimulated a xenoestrogenic response in cells, but such stimulus can be entirely attributable to the hydrolysis phenomenon, as the glycosylated form turned out to be unable to effectively bind and activate the estrogens receptors.

The parasitic mite Varroa destructor Anderson & Trueman (Acari: Mesostigmata) is the most challenging honey bee (Apis mellifera L.) pest for beekeepers worldwide. Studies involving the use of entomopathogenic fungi for control of mite populations have shown that there is potential for their use, but there are several obstacles to their direct application in the hive. One of the promising fungi is Metarrhizium anisopliae, which produces toxins involved in pathogenicity named “destruxins” (Dtx). In this study, we performed trials to evaluate the toxicity of crude and purified Dtx (fractions A, B, CE and D) towards V. destructor and A. mellifera. Mortality of mites treated with crude Dtx and Dtx B and CE was higher than control mites in all performed trials, in which several solvents and administration modes were employed. When purified Dtx B and CE were administered to honey bees at the higher concentration they caused a significantly higher mortality compared to control, showing the need for further research.

Coeliac disease is an autoimmune enteropathy that develops in genetically predisposed subjects after the ingestion of gluten or related proteins. Coeliac disease has an increasing incidence in the last years in western countries and it has been suggested that wheat breeding might have contributed to select more toxic forms of gluten. In this work, we analysed gluten peptides generated by in vitro digestion of different old and modern Triticum varieties, using LC-MS. We concluded that old varieties analysed produced a higher quantity of peptides containing immunogenic and toxic sequences than modern ones. Thus old wheat lines are not to be considered "safer" for subjects that are genetically predisposed to celiac disease.

The aim of this study examined the characterization of extra virgin olive oil samples from the main cultivar Chemlali, grown in five olive orchards with different soil type (Sandy, Clay, Stony, Brown, Limestone and Gypsum). Volatile compounds were studied using headspace-solid phase micro-extraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS) technics. Moreover, the sterol profile was established using gas chromatography-mass spectrometry. 35 different volatile compounds were identified: alcohols, esters, aldehydes, ketones and hydrocarbons. The chemical composition of the volatile fraction was characterized by the preeminence of 2-hexenal (32.75%) and 1-hexanol (31.88%). Three sterols were identified and characterized. For all olive oil samples, s-sitosterol (302.25 mg/kg) was the most abundant sterol. Interestingly, our results showed significant qualitative and quantitative differences in the levels of the volatile compounds and sterols from oils obtained from olive trees grown in different soil type.

Background Considering the large amounts of by-products derived from orange processing, which are generally discarded, the present study aimed to explore the feasibility of using orange peel for lactic acid production in solid state fermentation. Results Different species of lactic acid bacteria were employed, singly and in co-culture, to evaluate their ability to ferment orange peel and produce lactic acid. Among the single cultures tested, Lactobacillus casei 2246 was the most efficient strain, reaching the highest concentration of lactic acid (209.65 g kg-1 ) and yield (0.88 g g-1 ). The use of Lactobacillus plantarum 285 and Lactobacillus paracasei 4186 in co-culture produced a comparable amount of lactic acid, showing a better performance than the same strains in single cultures. Conclusion Orange peels represent a suitable raw material for solid state fermentation employing lactic acid bacteria. Lactic acid was obtained that consumed the most of sugars available, leading to high yields. Despite all the strains tested showing the same growth ability, different peculiarities in lactic acid production were revealed, dependent on the species/strains, suggesting the relevance of strain selection. © 2019 Society of Chemical Industry.

Deoxynivalenol (DON) and its modified forms (3-, and 15-acetyl-DON, DON-3-glucoside) are commonly analysed by chromatographic methods. Indeed, coupled with proper extraction and clean-up, LC-MS represents the best approach for multi-mycotoxin measurements. On the other hand, immunochemistry-based methods are possibly able to detect a family of structurally related compounds, although the determination of single contributions is not possible so far. However, ELISA methods often lead to an apparent overestimation of the mycotoxins content because modified forms and matrix components can potentially cross-react with the antibodies (designed for the parent toxin). Several data about the possible cross-reactivity of commercial DON-detecting ELISA kit are reported in the literature so far. Data are commonly obtained in buffer solutions or in matrix-matched solutions, but comparison of a set of naturally incurred samples has never been reported. In the present work the accuracy of a commercial DON-detecting ELISA kit was evaluated on naturally incurred soft wheat (n = 15) and maize (n = 15), taking into account the matrix effect. Recovery was calculated considering the DON concentration found by LC-MS/MS and the total DON concentration, expressed as the sum of DON and its modified forms found by LC-MS/MS. The obtained data clearly show that, when 3-modified forms of DON occur in the sample, the ELISA kit does actually detect them, thus returning an apparent overestimation if only DON content is considered. When the ELISA recovery is calculated on the total DON content, the accuracy of the analysis increases and the variability decreases. According to our data, the ELISA kit seems to be a promising group detection tool for the accurate evaluation of DON and its modified forms, expressed as sum of DON, DON-3Glc and 3Ac-DON, for soft wheat and maize samples.

This study investigated for the first time the intestinal transfer of two modified forms of zearalenone (ZEN), zearalenone-14-glucoside (ZEN14Glc) and zearalenone-16-glucoside (ZEN16Glc), using polarized monolayers of Caco-2 cells. The cells were apically exposed to 40 μM of ZEN14Glc, ZEN16Glc and ZEN, separately. Results showed that, after apical administration, ZEN14Glc and ZEN16Glc can be detected in cellular extracts indicating uptake by intestinal cells. Moreover, the glucosylated forms were cleaved to release ZEN, with a different cleavage ability for the two conjugated isomers. In particular, ZEN16Glc seems to be less prone to deglycosylation compared to ZEN14Glc, probably on account of an increased steric hindrance. We could show that human cytosolic β-glucosidase is able to cleave ZEN14Glc liberating ZEN, but is unable to cleave ZEN16Glc.All metabolites could cross the cell membrane and be detected in the apical compartment, while ZEN was also found in the basolateral compartment together with the modified mycotoxin form.

In the matter of foodborne mycotoxins, beside a number of regulated compounds, regulations are totally missing for phase-II plant metabolites--the toxicological knowledge of which is still in its infancy. Currently, zearalenone-14-glucoside is in the pipeline and its toxicological role is under a glowing scientific debate. In our work it clearly showed high toxicological concerns as it is prone to conversion to well-known toxic compounds (i.e. zearalenone and both zearalenol isomers) when exposed to breast cancer cells culture. The need of future risk assessment studies has been pointed out accordingly.

Worldwide, thousands of insect species are consumed as food or are used as feed ingredients. Hermetia illucens, &lsquo, black soldier fly&rsquo, is one of them, and a large amount of puparia and dead adults flies are accumulated during rearing. These materials represent important wastes but no studies are still present in the literature regarding their functional properties and potential reuse. Lactic acid bacteria (LAB) are a heterogeneous group of bacteria contributing to various industrial applications, ranging from food fermentation, chemicals production to pharmaceuticals manufacturing. A LAB feature of industrial interest is their ability to produce antimicrobial metabolites. Considering the scientific and commercial interest in discovering novel antimicrobials, this work will be direct towards fermentation of insect-derived biomasses: puparia and adults insect at the end of life cycle. To the best of our knowledge, the in vitro antimicrobial activity of fermented insects is tested for the first time. This study aimed also to evaluate differences in the composition between fermented and unfermented insects, and to study whether the fermentation and the type of LAB used played a crucial role in modifying the composition of the substrate. Results firstly highlighted fermentability of this species of insects, showed that fermented black soldier flies puparium possess a high antimicrobial activity against tested pathogens. Moreover, result of chemical composition showed that fermented biomass had a higher percentage of fat and a more complex fatty acids profile.

Emodin, a widespread natural anthraquinone, has many biological activities including health-protective and adverse effects. Amongst beneficial effects, potential antiviral activity against coronavirus responsible for the severe acute respiratory syndrome outbreak in 2002&ndash, 2003 has been described associated with the inhibition of the host cells target receptors recognition by the viral Spike protein. However, the inhibition mechanisms have not been fully characterized, hindering the rational use of emodin as a model compound to develop more effective analogues. This work investigates emodin interaction with the Spike protein to provide a mechanistic explanation of such inhibition. A 3D molecular modeling approach consisting of docking simulations, pharmacophoric analysis and molecular dynamics was used. The plausible mechanism is described as an interaction of emodin at the protein&ndash, protein interface which destabilizes the viral protein-target receptor complex. This analysis has been extended to the Spike protein of the coronavirus responsible for the current pandemic hypothesizing emodin&rsquo, s functional conservation. This solid knowledge-based foothold provides a possible mechanistic rationale of the antiviral activity of emodin as a future basis for the potential development of efficient antiviral cognate compounds. Data gaps and future work on emodin-related adverse effects in parallel to its antiviral pharmacology are explored.

The xenoestrogenic mycotoxin zearalenone is a common food contaminant produced by Fusarium strains. The modified mycotoxin zearalenone-14-sulfate (Z14S) is formed in both fungi and mammals during the biotransformation of zearalenone. Cyclodextrins (CD) are cyclic oligosaccharides which can form host-guest type complexes with some mycotoxins, including zearalenone, zearalenols, and zearalenone-14-glucoside. As a result of the complex formation, the fluorescence signal of these mycotoxins strongly increases. Furthermore, CD polymers seem to be suitable for the extraction of some mycotoxins from aqueous solutions and beverages. In this study, the interaction of Z14S with CDs and soluble CD polymers was examined with fluorescence spectroscopy and molecular modeling. Furthermore, the removal of Z14S from aqueous solution by β-CD bead polymer (BBP) was also tested. Our results demonstrate the formation of stable Z14S-CD complexes (K = 0.1 to 5.0 × 104 L/mol). Dimethyl-β-CD (DIMEB) produced the most stable complexes with Z14S at pH 5.0 and 7.4. At pH 10.0, the binding constant of Z14S-DIMEB complex decreased and quaternary ammonium-β-CD showed similar affinity toward the mycotoxin than DIMEB. In addition, Z14S was successfully removed from aqueous solutions by BBP. Considering the above-listed observations, besides the parent mycotoxins, some of their modified/masked derivatives can also interact with CDs.

Wheat (Triticum spp. L.) is an important source of nutrients and bioactive compounds with recognized beneficial effects. Wheat undergoes several processes with the final aim of separating the endosperm from the outer layers, usually discarded. In this study, free and bound phenolic acids (PAs) profile, betaine and choline contents were quantified in six different wheat species (durum and bread wheat, turanicum wheat, einkorn, emmer and spelt), the corresponding milling by-products (bran, middlings, aleurone and I, II and III steps of debranning) and flour/semolina, using UHPLC-MS/MS methods. The bound form of phenolics was the component present in higher concentration (80% of the total, in average) and ferulic acid was the most abundant compounds, representing between 67 and 73 % of total PAs. Among the species, bread wheat grain totalized the highest content of total PAs (1209.31 ± 7.3 µg g−1 d.w.). Betaine and choline are abundantly present in wheat species. In general, the highest content of bioactive compounds was found in bran (3 times higher than whole grains), emphasizing the good nutritional profile of these by-products. The milling process leads to a severe reduction of phenolic acids and methyl-donors in the end-products.

This study aimed to investigate the potential of in vitro wheat model as biofactory for masked mycotoxin production. Micropropagated durum wheat organs (leaves and roots) were treated during a 14-day time span on a proper medium spiked with deoxynivalenol (DON). After the treatment, DON absorption from culture media was evaluated while roots and leaves were profiled by UHPLC-HRMS to investigate the DON biotransformation products. A total of 10 metabolites have been annotated in both roots and leaves. In particular, 5 phase I metabolites never reported before were putatively identified, suggesting the viability of the model as a tool to investigate the interplay between mycotoxins and wheat. In addition, 5 phase II metabolites previously reported in wheat grown under open field conditions, were identified in both roots and leaves, thus demonstrating the reliability of the cultured organs as model system for wheat plants. An organ-dependent difference in DON uptake and biotransformation was observed, since roots contained a high amount of untransformed DON, while leaves were able to effectively biotransform DON to its glycosylated form and other relevant metabolites. With the perspective of using cultured organs as biofactories for modified mycotoxin production, leaves seemed therefore to offer the best absorption and production yield.