Nano-Hybrid Ag@LCCs Systems with Potential Wound-Healing Properties.

The synthesis of contaminant-free silver@linear carbon chains (Ag@LCCs) nanohybrid systems, at different Ag/LCCs ratios, by pulsed laser ablation was studied. The ablation products were first characterized by several diagnostic techniques: conventional UV-Vis optical absorption and micro-Raman spectroscopies, as well as scanning electron microscopy, operating in transmission mode. The experimental evidence was confirmed by the theoretical simulations' data. Furthermore, to gain a deeper insight into the factors influencing metal@LCCs biological responses in relation to their physical properties, in this work, we investigated the bioproperties of the Ag@LCCs nanosystems towards a wound-healing activity. We found that Ag@LCC nanohybrids maintain good antibacterial properties and possess a better capability, in comparison with Ag NPs, of interacting with mammalian cells, allowing us to hypothesize that mainly the Ag@LCCs 3:1 might be suitable for topical application in wound healing, independent of (or in addition to) the antibacterial effect.

The p-Phthalates Terephthalic Acid and Dimethyl Terephthalate Used in the Manufacture of PET Induce In Vitro Adipocytes Dysfunction by Altering Adipogenesis and Thermogenesis Mechanisms.

Public health concerns associated with the potential leaching of substances from Polyethylene terephthalate (PET) packaging have been raised due to the role of phthalates as endocrine-disrupting chemicals or obesogens. In particular, changes in the environment such as pH, temperature, and irradiation can improve contaminant migration from PET food packaging. In this study, the in vitro effects of p-phthalates terephthalic acid (TPA) and dimethyl terephthalate (DMT) on murine adipocytes (3T3-L1) were evaluated using concentrations that might be obtained in adult humans exposed to contaminated sources. TPA and, in particular, DMT exposure during 3T3-L1 differentiation increased the cellular lipid content and induced adipogenic markers PPAR-γ, C/EBPß, FABP4, and FASN, starting from low nanomolar concentrations. Interestingly, the adipogenic action of TPA- and DMT-induced PPAR-γ was reverted by ICI 182,780, a specific antagonist of the estrogen receptor. Furthermore, TPA and DMT affected adipocytes' thermogenic program, reducing pAMPK and PGC-1α levels, and induced the NF-κB proinflammatory pathway. Given the observed effects of biologically relevant chronic concentrations of these p-phthalates and taking into account humans' close and constant contact with plastics, it seems appropriate that ascertaining safe levels of TPA and DMT exposure is considered a high priority.

Black Soldier Fly Larvae Meal in the Diet of Gilthead Sea Bream: Effect on Chemical and Microbiological Quality of Filets.

The chemical and microbiological characteristics of filets of Spaurus aurata L. specimens fed with diets containing a Hermetia illucens meal (HIM) at the 25, 35, and 50%, as a partial replacement for fish meal (FM) were evaluated. The diets, formulated to satisfy the nutritional needs of fish, were isoenergetic (22 MJ/kg gross energy), isonitrogenous (43 g/100 g, a.f.), and isolipidic (19 g/100 g, a.f.). Seventy-two specimens were randomly killed after 186 days of growing trials. Then, the filets were analyzed for chemical profile, fatty acids, amino acids, minerals, and microbial flora. Data were subjected to statistical analysis. No significant differences were observed in chemical composition. The sum of polyunsaturated fatty acids (PUFAs) showed a similar content in the filets; eicosapentaenoic acid was similar in the filets of HIM0, HIM35%, and HIM50%, whereas docosahexaenoic acid was higher in filets of the HIM0 group. n3/n6 PUFA ratio and the sum of EPA + DHA showed a high value (p < 0.001) in filets of the group fed with FM. No significant difference was observed in thrombogenic index and hypocholesterolaemic/hypercholesterolaemic ratio in the groups; the atherogenic index showed a higher value (p = 0.001) in the HIM50% group. Indispensable amino acids showed some significant (p < 0.0001) differences in the groups; arginine and phenylalanine content was higher in the filets of fish fed with FM; isoleucine and valine content was higher in the filets of HIM50%; leucine, lysine and methionine content was lower in the filets of HIM35%; histidine content was lower in the filets of HIM25%; tryptophan content was lower in filets of the HIM50% group. EAA/NEAA ratio showed highest value in the filets of the group that received FM. The presence of HIM in the three diets kept chromium, manganese, iron, copper, zinc, and nickel levels lower than those recommended by various authorities. Ca/P ratio showed a higher level (p < 0.0001) in the group fed with FM than those fed with diets containing HIM. The insect meal in the diets did not influence the microbiological profile of fish. Use of HIM as an unconventional feed ingredient in Sparus aurata diet looks promising, although the quality of filets may be affected.

Antibacterial Nanoassembled Calix[4]arene Exposing Choline Units Inhibits Biofilm and Motility of Gram Negative Bacteria.

The high incidence of antibiotic resistance and biofilm-associated infections is still a major cause of morbidity and mortality and triggers the need for new antimicrobial drugs and strategies. Nanotechnology is an emerging approach in the search for novel antimicrobial agents. The aim of this study was to investigate the inherent antibacterial effects of a self-assembling amphiphilic choline-calix[4]arene derivative (Chol-Calix) against Gram negative bacteria. Chol-Calix showed activity against Escherichia coli and Pseudomonas aeruginosa, including antibiotic-resistant strains, and affected the bacterial biofilm and motility. The activity is likely related to the amphipathicity and cationic surface of Chol-Calix nanoassembly that can establish large contact interactions with the bacterial surface. Chol-Calix appears to be a promising candidate in the search for novel nanosized nonconventional antimicrobials.

In Vitro Protective Effects of a Standardized Extract From Cynara Cardunculus L. Leaves Against TNF-α-Induced Intestinal Inflammation.

Inflammatory bowel disease (IBD) represents a group of progressive disorders characterized by recurrent chronic inflammation of the gut. New unconventional therapies based on plant derived compounds capable of preventing and/or reducing acute or chronic inflammation could represent a valid alternative for the treatment or prevention of IBDs. Cynara cardunculus L. leaves, considered a food-waste suitable as a rich source of bioactive polyphenols including luteolin and chlorogenic acid, has been reported for its positive effects in digestive tract. The aim of the present work was to evaluate the in vitro molecular mechanisms of beneficial effects of a standardized polyphenol-rich extract obtained from the leaves of Cynara cardunculus L (CCLE) against acute intestinal inflammation induced by TNF-α on intestinal epithelial Caco-2 cells. CCLE prevented TNF-α-induced NF-κB inflammatory pathway and the overexpression of IL-8 and COX-2. In addition, CCLE was able to improve basal intracellular antioxidant power in both TNF-α-unexposed or -exposed Caco-2 cells and this effect was associated to the activation of Nrf2 pathway, a master regulator of redox homeostasis affecting antioxidant and phase II detoxifying genes, stimulating an adaptive cellular response. In conclusion, our data clearly evidenced that, although considered a waste, Cynara cardunculus leaves may be used to obtain extracts rich in bioactive polyphenols potentially useful for prevention and treatment of inflammatory intestinal diseases.

Grape water: reclaim and valorization of a by-product from the industrial cryoconcentration of grape (Vitis vinifera) must.

BACKGROUND: The term 'grape (Vitis vinifera) water' refers to a by-product from the cryoconcentration of must that, if not reclaimed, would be considered as wastewater. In this study, the nutraceutical potential of waters reclaimed from the cryoconcentration of Grillo and Moscato musts was evaluated. RESULTS: Both waters showed physicochemical parameters in agreement with Italian regulation for drinking water, and interesting levels of F- (3.02-8.02 mg L-1 ) and SO4 - (52.85-49.34 mg L-1 ). Inorganic elements, including Mg (5.54-7.78 mg L-1 ), K (47.12-59.87 mg L-1 ), Fe (219.09-205.32 µg L-1 ), and Zn (189.65-127.30 µg L-1 ), and phenolic contents <35 mg GAE L-1 contributed to determine moderate antioxidant activities. Considering fatty acid composition, oleic and linoleic acids predominated, being higher in Moscato than Grillo samples (64.42% versus 58.22%, and 5.42% versus 6.07%). Grape waters displayed also rich aroma profiles, including mainly esters, alcohols and terpenoids. The latter components (i.e. linalool and α-terpineol) were more abundant in Moscato than in Grillo (13% versus 8%). Interestingly, some minor volatiles, characterized the vine of provenance (e.g. benzaldehyde and 3-methylbutylacetate in Moscato). All grape waters showed also a considerable fraction of ethyl lactate, six-carbon compounds and acetates, reflecting the proximity of samples to the grapes of origin. CONCLUSION: In a growing scenario of environmental decay and resource depletion, results from this study support an innovative and profitable waste recycling strategy for the wine industry. © 2020 Society of Chemical Industry.

Nitrogen Headspace Improves the Extra Virgin Olive Oil Shelf-Life, Preserving Its Functional Properties.

The functional foods field has recently evolved due to new research being carried out in the food area and greater regulations; these factors have contributed to the creation of health claims, and to the increasing attention that consumers give to health-promoting food products. The aim of this research was to improve the shelf-life of a typical functional food of the Mediterranean diet, the Extra Virgin Olive Oil (EVOO). We focused our attention on the standardization and validation of a production process, starting from the cultivation and harvesting of the olives, which would guarantee a product of quality in terms of bioactive compound content. Furthermore, a methodology/procedure to preserve them in the best way over a long period of time, in order to guarantee the consumer receives a product that retains its functional and organoleptic native properties, was evaluated. The monitoring of biological cultivations, harvesting, milling process, and storage, as well as careful quality control of the analytical parameters (e.g., contents of polyphenols, α-tocopherol, fatty acids, acidity, peroxides, dienes, trienes, ΔK, and antioxidant power) showed that, under the same conditions, a nitrogen headspace is a discriminating factor for the maintenance of the functional properties of EVOO.

Analytical Evaluation and Antioxidant Properties of Some Secondary Metabolites in Northern Italian Mono- and Multi-Varietal Extra Virgin Olive Oils (EVOOs) from Early and Late Harvested Olives.

The antioxidant activity and the phenolic and α-tocopherol content of 10 Northern Italian mono- and multi-varietal extra virgin olive oils (EVOOs), after early and late olive harvests, was analyzed. A hierarchical cluster analysis was used to evaluate sample similarity. Secoiridoids (SIDs), lignans and flavonoids were the most abundant phenolic compounds identified. The organic Casaliva (among mono-cultivar) and the organic multi-varietal (among blended oils) EVOOs had the higher total phenol content both in early (263.13 and 326.19 mg/kg, respectively) and late harvest (241.88 and 292.34 mg/kg, respectively) conditions. In comparison to late harvest EVOOs, early harvest EVOOs, in particular the organic mono-cultivar Casaliva, showed both higher antioxidant capacity (up to 1285.97 Oxygen Radicals Absorbance Capacity/ORAC units), probably due to the higher SID fraction (54% vs. 40%), and higher α-tocopherol content (up to 280.67 mg/kg). Overall, these results suggest that SIDs and α-tocopherol mainly contribute to antioxidant properties of the studied EVOOs. In light of this, the authors conclude that early harvest, organic mono-cultivar Casaliva EVOO represents the most interesting candidate to explicate healthy effects ascribed to these functional constituents, particularly regarding oxidative stress-related pathologies.

Copper chelation and interleukin-6 proinflammatory cytokine effects on expression of different proteins involved in iron metabolism in HepG2 cell line.

BACKGROUND: In vertebrates, there is an intimate relationship between copper and iron homeostasis. Copper deficiency, which leads to a defect in ceruloplasmin enzymatic activity, has a strong effect on iron homeostasis resulting in cellular iron retention. Much is known about the mechanisms underlying cellular iron retention under "normal" conditions, however, less is known about the effect of copper deficiency during inflammation. RESULTS: We show that copper deficiency and the inflammatory cytokine interleukin-6 have different effects on the expression of proteins involved in iron and copper metabolism such as the soluble and glycosylphosphtidylinositol anchored forms of ceruloplasmin, hepcidin, ferroportin1, transferrin receptor1, divalent metal transporter1 and H-ferritin subunit. We demonstrate, using the human HepG2 cell line, that in addition to ceruloplasmin isoforms, copper deficiency affects other proteins, some posttranslationally and some at the transcriptional level. The addition of interleukin-6, moreover, has different effects on expression of ferroportin1 and ceruloplasmin, in which ferroportin1 is decreased while ceruloplasmin is increased. These effects are stronger when a copper chelating agent and IL-6 are used simultaneously. CONCLUSIONS: These results suggest that copper chelation has effects not only on ceruloplasmin but also on other proteins involved in iron metabolism, sometimes at the mRNA level and, in inflammatory conditions, the functions of ferroportin and ceruloplasmin may be independent.

The interaction and binding of flavonoids to human serum albumin modify its conformation, stability and resistance against aggregation and oxidative injuries.

BACKGROUND: Interactions of ligands with proteins imply changes in the properties of the macromolecules that may deeply modify their biological activities and conformations and allow them to acquire new and, sometimes, unexpected abilities. The flavonoid phloretin has several pharmacological properties that are starting to be elucidated, one of which is the well-known inhibition of glucose transport. METHODS: The interactions of phloretin to human serum albumin have been investigated by fluorescence, UV-visible, FTIR spectroscopy, native electrophoresis, protein ligand docking studies, fluorescence and scanning electron microscopy. RESULTS: Spectroscopic investigations suggest that the flavonoid binds to human serum albumin inducing a decrease in α-helix structures as shown by deconvolution of FTIR Amide I' band. Fluorescence and displacement studies highlight modifications of environment around Trp214 with the primary binding site located in the Sudlow's site I. In the hydrophobic cavity of subdomain IIA, molecular modeling studies suggest that phloretin is in non-planar conformation and hydrogen-bonded with Ser202 and Ser454. These changes make HSA able to withstand protein degradation due to HCLO and fibrillation. GENERAL SIGNIFICANCE: Our work aims to open new perspectives as far as the binding of flavonoids to HSA are concern and shows as the properties of both compounds can be remarkable modified after the complex formation, resulting, for instance, in a protein structure much more resistant to oxidation and fibrillation. This article is part of a Special Issue entitled "Science for Life" Guest Editor: Dr. Austen Angell, Dr. Salvatore Magazù and Dr. Federica Migliardo.

Antiepileptic carbamazepine drug treatment induces alteration of membrane in red blood cells: possible positive effects on metabolism and oxidative stress.

Carbamazepine (CBZ) is an iminostilbene derivative commonly used for treatment of neuralgic pain and bipolar affective disorders. CBZ blood levels of treated patients are within the range of micromolar concentrations and therefore, significant interactions of this drug with erythrocytes are very likely. Moreover, the lipid domains of the cell membrane are believed to be one of the sites where iminostilbene derivatives exert their effects. The present study aimed to deeply characterize CBZ effects on erythrocytes, in order to identify extra and/or cytosolic cell targets. Our results indicate that erythrocyte morphological changes promoted by the drug, may be triggered by an alteration in band 3 functionality i.e. at the level of anionic flux. In addition, from a metabolic point of view this perturbation could be considered, at least in part, as a beneficial event because it could favour the CO2 elimination. Since lipid peroxidation, superoxide and free radical scavenging activities, caspase 3 activity and hemoglobin (Hb) functionality were not modified within the CBZ treated red blood cell (RBC), band 3 protein (B3) may well be a specific membrane target for CBZ and responsible for CBZ-induced toxic effects in erythrocytes. However some beneficial effects of this drug have been evidenced; among them an increased release of ATP and nitric oxide (NO) derived metabolites from erythrocytes to lumen, leading to an increased NO pool in the vasculature. In conclusion, these results indicate that CBZ, though considered responsible for toxic effects on erythrocytes, can also exhibit effects that at least in some conditions may be seen as beneficial.

Flavonoid glycosides in bergamot juice (Citrus bergamia Risso).

A comprehensive profile of flavonoids in bergamot juice was obtained by a single DAD-ESI-LC-MS-MS course. Eight flavonoids were found for the first time, five of these are C-glucosides (lucenin-2, stellarin-2, isovitexin, scoparin, and orientin 4'-methyl ether), and three are O-glycosides (rhoifolin 4'-O-glucoside, chrysoeriol 7-O-neohesperidoside-4'-O-glucoside, and chrysoeriol 7-O-neohesperidoside). A method is proposed to differentiate chrysoeriol and diosmetin derivatives, which are often indistinguishable by LC-MS-MS. In-depth knowledge of the flavonoid content is the starting point for bergamot juice exploitation in food industry applications.

Flavonoids detection by HPLC-DAD-MS-MS in lemon juices from Sicilian cultivars.

High-performance liquid chromatography-diode array detector (HPLC-DAD) coupled with electronspray mass spectrometry (ESI-MS) was used to detect the flavonoid profile in lemon juices obtained from the main Sicilian cultivars (Femminello comune, Monachello, and Interdonato). Significant amounts of an unusual constituent were found in the lemon juice of the above-mentioned cultivars together with eriocitrin, hesperidin, and diosmin. Following purification by preparative HPLC, the structure was assigned as 6,8-di-C-glucopyranosyldiosmetin by means of DAD-UV, ESI-MS-MS, and (1)H NMR analysis. Three other minor components were also detected. One of these presented a flavone nature, and spectral data and literature references both suggested a 6,8-di-C-glucopyranosylapigenin structure. The different contents of eriocitrin, hesperidin, diosmin, and above minor components in the cultivars allow juices to be readily differentiated.