Effects of Electron Beam Radiation on the Phenolic Composition and Bioactive Properties of Olive Pomace Extracts.

Olive pomace is an agro-industrial waste product generated from the olive oil industry and constituted by bioactive compounds with potential applications in several industrial sectors. The purpose of this work was to evaluate the effects of electron beam (e-beam) radiation on olive pomace, specifically on phenolic compounds (by HPLC-DAD-ESI/MS) and the bioactive properties (antioxidant, antiproliferative, and antimicrobial activities) of crude olive pomace (COP) and extracted olive pomace (EOP) extracts. The amount of total flavonoid content and the reducing power of COP extracts were higher than those obtained for EOP extracts. The results suggested that e-beam radiation at 6 kGy increased both total phenolic and total flavonoid contents as well as the reducing power of COP extracts, due to the higher extractability (>2.5-fold) of phenolic compounds from these samples, while decreasing the scavenging activity of extracts. The extracts of both olive pomaces showed antibacterial potential, and COP extracts at 400 µg/mL also presented antiproliferative activity against A549, Caco-2, 293T, and RAW264.7 cell lines, with both properties preserved with the e-beam treatment. All in all, e-beam radiation at 6 kGy appears to be a promising technology to valorize the pollutant wastes of the olive oil industry through enhancing phenolic extractability and bioactive properties, and, furthermore, to contribute to the environmental and economical sustainability of the olive oil industry.

Insights into the Neuroprotective Potential of Epicatechin: Effects against Aß-Induced Toxicity in Caenorhabditis elegans.

Medical therapies to avoid the progression of Alzheimer's disease (AD) are limited to date. Certain diets have been associated with a lower incidence of neurodegenerative diseases. In particular, the regular intake of foods rich in polyphenols, such as epicatechin (EC), could help prevent or mitigate AD progression. This work aims to explore the neuroprotective effects of EC using different transgenic strains of Caenorhabditis elegans, which express human Aß1-42 peptides and contribute to elucidating the mechanisms involved in the effects of EC in AD. The performed assays indicate that this flavan-3-ol was able to reduce the signs of ß-amyloid accumulation in C. elegans, improving motility and chemotaxis and increasing survival in transgenic strain peptide producers compared to nematodes not treated with EC. The neuroprotective effects exhibited by EC in C. elegans could be explained by the modulation of inflammation and stress-associated genes, as well as autophagy, microgliosis, and heat shock signaling pathways, involving the regulation of cpr-5, epg-8, ced-7, ZC239.12, and hsp-16 genes. Overall, the results obtained in this study support the protective effects of epicatechin against Aß-induced toxicity.

Magnesium and manganese induced changes on chemical, nutritional, antioxidant and antimicrobial properties of the pansy and Viola edible flowers.

Pansy and viola edible flowers were grown hydroponically with different levels of Mg and Mn. The nutritional composition was determined using standard methods. Free sugars, fatty acids, organic acids, tocopherols, and phenolic compounds were analyzed using various HPLC and GC devises. The extract's antimicrobial, antioxidant, cytotoxicity, and anti-inflammatory activity were assessed. The results indicated that Mg enrichment negatively affected plant growth and mineral accumulation but improved photosynthetic performance. The edible flowers contained significant amounts of protein, low levels of fat, and varying sugar contents, such as glucose and fructose. Various fatty acids and phenolic compounds were identified, with different concentrations depending on the treatment. The flowers exhibited antioxidant potential, antimicrobial activity, cytotoxic effects, and anti-inflammatory properties. The correlations between the investigated parameters not only expand knowledge on Mg and Mn interaction but also catalyze significant advancements in sustainable agriculture and food health, fostering a healthier and more conscious future.

Microbiota Assessment of Fresh-Cut Apples Packaged in Two Different Films.

The aim of this work was to assess the natural microbiota of packed fresh-cut apples during refrigerated storage. Two different films were tested for the package, a biodegradable (PLA) film and a conventional and commercial one (OPP). Two antioxidant additives were applied, a natural olive pomace extract and the commercial ascorbic acid used by the industries. The results revealed lower bacteria counts in samples with olive pomace extract and PLA films than in those with ascorbic acid and OPP films after 5 and 12 days of storage. These findings suggest that the use of such natural extracts as additives in fruits could delay the growth of mesophilic bacteria. The characterization and identification of the bacterial isolates from fresh-cut apple samples showed that the most prevalent species were Citrobacter freundii, Staphylococcus warneri, Pseudomonas oryzihabitans, Alcalinogenes faecalis, Corynebacterium jeikeium, Micrococcus spp., Pantoea aglomerans and Bacillus spp. Furthermore, an increase in the microbial diversity during the storage time at refrigerated temperatures was observed, except for the sample treated with olive pomace extract and packaged in OPP film. The highest microbial diversity was found for samples with ascorbic acid as an additive. This could indicate a negative effect of ascorbic acid on the microbial inhibition of apple slices. The natural olive pomace extract demonstrated potential as an antimicrobial additive for fresh-cut apples.

Effects of the extraction techniques on the chemical composition and bioactive properties of lemon balm (Melissa officinalis L.) plants grown under different cropping and irrigation regimes.

The present study aims to determine the combined effect of cropping system and irrigation regime on the chemical composition and bioactive properties of lemon balm aerial parts. For this purpose, lemon balm plants were grown under two farming systems (conventional farming vs organic farming) and two irrigation levels (full irrigation vs deficit irrigation) and harvested twice throughout the growing period. The collected aerial parts were subjected to three different methods of extractions, namely infusion, maceration and ultrasound-assisted extraction and the obtained extracts were evaluated in terms of chemical profile and bioactivities. Five organic acids with varied composition among the tested treatments were identified in all the tested samples for both harvests, namely, citric, malic, oxalic, shikimic and quinic acid. Regarding phenolic compounds composition, the most abundant ones were rosmarinic acid, lithospermic acid A isomer I and hydroxylsalvianolic E, especially for the maceration and infusion extraction methods. Full irrigation resulted in lower EC50 values than deficit irrigation only in the treatments of the second harvest, while variable cytotoxic and anti-inflammatory effects were recorded in both harvests. Finally, in most cases the lemon balm extracts has similar or better activity than the positive controls, while the antifungal activity of lemon balm extracts was stronger than the antibacterial effects. In conclusion, the results of the present study showed that the implemented agronomic practices, as well as the extraction protocol may significantly affect the chemical profile and bioactivities of lemon balm extracts, suggesting that both the farming system and the irrigation schedule may improve the quality of the extracts depending on the implemented extraction protocol.

The wide spectrum of industrial applications for cultivated cardoon (Cynara cardunculus L. var. Altilis DC.): A review.

Cynara cardunculus L. var. altilis DC. belongs to the Asteraceae family and is widely used. This species is integrated into the Mediterranean diet and has broad applicability due to its rich chemical composition. Its flowers, used as a vegetable coagulant for gourmet cheese production, are rich in aspartic proteases. Leaves are rich in sesquiterpene lactones, the most abundant being cynaropicrin, while stems present a higher abundance of hydroxycinnamic acids. Both classes of compounds exhibit a wide range of bioactive properties. Its chemical composition makes it applicable in other industrial sectors, such as energy (e.g., manufacturing of biodiesel and biofuel) or paper pulp production, among other biotechnological applications. In the last decade, cardoon has been identified as a competitive energy crop, constituting an opportunity for the economic recovery and development of the rural areas of the Mediterranean basin. This article reviews the chemical composition, bioactive properties, and multifaceted industrial applications of cardoon.

Effect of Olive Pomace Extract Application and Packaging Material on the Preservation of Fresh-Cut Royal Gala Apples.

The efficiency of natural olive pomace extracts for enhancing the quality of fresh-cut apples was compared with commercial ascorbic acid and two different packaging films (biodegradable polylactic acid (PLA) and oriented polypropylene (OPP)) were tested. The composition of atmosphere inside the packages, the physicochemical parameters (firmness, weight loss and color), the microbial load, total phenolic content and antioxidant activity of fresh-cut apples were evaluated throughout 12 days of storage at 4 °C. After 12 days of refrigerated storage, a significant decrease in O2 was promoted in PLA films, and the weight loss of the whole packaging was higher in PLA films (5.4%) than in OPP films (0.2%). Natural olive pomace extracts reduced the load of mesophilic bacteria (3.4 ± 0.1 log CFU/g and 2.4 ± 0.1 log CFU/g for OPP and PLA films, respectively) and filamentous fungi (3.3 ± 0.1 log CFU/g and 2.44 ± 0.05 log CFU/g for OPP and PLA films, respectively) growth in fresh-cut apples after five days of storage at 4 °C, and no detection of coliforms was verified throughout the 12 days of storage. In general, the olive pomace extract preserved or improved the total phenolic index and antioxidant potential of the fruit, without significant changes in their firmness. Moreover, this extract seemed to be more effective when combined with the biodegradable PLA film packaging. This work can contribute to the availability of effective natural food additives, the sustainability of the olive oil industries and the reduction of environmental impact. It can also be useful in meeting the food industries requirements to develop new functional food products.

Ultrasound-assisted extraction of hydroxytyrosol and tyrosol from olive pomace treated by gamma radiation: process optimization and bioactivity assessment.

Ultrasound-assisted extraction (UAE) was used to recover hydroxytyrosol and tyrosol from olive pomace, a residue generated by the olive oil industry. The extraction process was optimized using response surface methodology (RSM), with processing time, ethanol concentration and ultrasonic power as the combined independent variables. The highest amounts of hydroxytyrosol (36 ± 2 mg g-1 of extract) and tyrosol (14 ± 1 mg g-1 of extract) were obtained after 28 min of sonication at 490 W using 7.3% ethanol as the solvent. Under these global conditions, an extraction yield of 30 ± 2% was achieved. The bioactivity of the extract obtained under optimized UAE was evaluated and compared with that of an extract obtained under optimal heat-assisted extraction (HAE) conditions in a previous work of the authors. Compared to HAE, UAE reduced the extraction time and the solvent consumption, and also led to higher extraction yields (HAE yield was 13.7%). Despite this, HAE extract presented higher antioxidant, antidiabetic, anti-inflammatory and antibacterial activities and no antifungal potential against C. albicans. Furthermore, HAE extract also showed higher cytotoxic effects against the breast adenocarcinoma (MCF-7) cell line. These findings provide useful information for the food and pharmaceutical industries in developing new bioactive ingredients, which may represent a sustainable alternative to synthetic preservatives and/or additives.

The effects of polyphenols against oxidative stress in Caenorhabditis elegans are determined by coexisting bacteria.

Introduction: Increasing evidence supports the role of gut microbiota in many aspects of human health, including immune, metabolic and neurobehavioral traits. Several studies have focused on how different components of the diet, such as polyphenols, can modulate the composition and function of the gut microbiota leading to health benefits. Methods: The effects on the resistance against thermally induced oxidative stress of C. elegans grown in the presence of flavonoids (quercetin or epicatechin) and fed different probiotic strains, namely Lactobacillus plantarum CLC17, Bifidobacterium longum NCIMB 8809 and Enterococcus faecium CECT 410, were explored. Results: Feeding C. elegans with the assayed bacteria in the absence of flavonoids did not significantly affect body size and fertility of the worms neither improve their resistance against oxidative stress compared to E. coli controls. However, increased resistance to stress was found when C. elegans was cultivated in the presence of both L. plantarum and flavonoids, but not with B. longum or E. faecium. An exploratory study revealed the presence of glycosylated and sulfated metabolites together with the aglycone in worms treated with quercetin and fed any of the different assayed LAB strains. However, in the assays with epicatechin a differential metabolite, tentatively identified as 5-(4'-hydroxyphenyl)-γ-valerolactone 3'-O-glucoside, was detected in the worms fed L. plantarum but not with the other bacteria. Conclusion: The obtained results indicated that the interactions bacteria/polyphenol play a key role in the effects produced in C. elegans regarding resistance against oxidative stress, although those effects cannot be only explained by the ability of bacteria to metabolize polyphenols, but other mechanisms should also be involved.

Effect of Polyphenolic Complements on Cognitive Function in the Elderly: A Systematic Review.

Polyphenols have been shown to be effective against many chronic diseases. These compounds could have a beneficial effect at the cognitive level. The exact mechanism by which they provide positive effects at the cognitive level is not well known, but it is believed that they could counteract neuroinflammation. The objective of this study is to review nutritional interventions that include foods or supplements rich in flavanols, flavonols, or stilbenes to the usual diet on cognitive deterioration in people over 50 years of age. Clinical trials published in PubMed and Web of Science from 1 March 2010 to 1 March 2020 were explored, from which 14 studies were selected. All of them showed some improvement after the intervention. In interventions with flavanols and stilbenes, relevant improvements have been observed both in healthy patients and in patients with established cognitive impairment. Most studies agree that the greatest benefits are found with high doses and longer duration treatments. The changes were fundamentally assessed through cognitive tests, and in some of the studies, through magnetic resonance imaging (fMRI). The type of cognitive test used to assess the effect of the intervention was revealed to be critical. Several studies have also shown improvements in analytical parameters and blood pressure.

Chemical composition of cardoon (Cynara cardunculus L. var. altilis) petioles as affected by plant growth stage.

In the present study petioles were collected from cultivated cardoon (Cynara cardunculus L. var altilis) plants in central Greece at sixteen harvest dates (samples P1-P16) and nine growth stages (principal growth stages (PGS) 1-9). The main objective of this study was to study the influence of petioles' growth cycle on their chemical composition. The lipidic content, fatty acids, free sugars, organic acids, and tocopherol composition were analyzed. A total of 27 fatty acids were identified, being the palmitic acid (C16:0, 12.42-50%) the most abundant, except for samples P4 and P5 where linoleic acid revealed the highest relative abundance (C18:2n6c, 46.5 and 39.05%, respectively). The α-, ß-, and γ-tocopherols were the identified isoforms of vitamin E, with sample P1 recording the highest tocopherol content. The detected organic acids were oxalic, quinic, malic, citric, and fumaric acids with the highest content being observed in sample P1. Sucrose was the major sugar in all the analyzed samples, followed by fructose, glucose, trehalose, and raffinose. The obtained results allowed us to characterize the chemical composition of cardoon petioles throughout its growth cycle and, consequently, to valorize the species through the exploitation of underutilized plant tissues.

Phenolic Composition and Antioxidant, Anti-Inflammatory, Cytotoxic, and Antimicrobial Activities of Cardoon Blades at Different Growth Stages.

Cardoon (Cynara cardunculus var. altilis) blades were collected at sixteen sampling dates (B1-B16) to study the influence of the phenological growth stage on the phenolic composition and biological properties. Twenty phenolic compounds were identified, among which trans 3,4-O-dicaffeoylquinic acid, 5-O-caffeoylquinic acid, and luteolin-O-hexoside (39.6, 42.6, and 101.0 mg/g extract, respectively) were the main compounds. Immature blades (B3) had a higher content of phenolic compounds (178 mg/g extract) and a greater ability to inhibit the formation of thiobarbituric acid reactive substances (IC50 of 1.61 µg/mL). Samples at more advanced growth stages revealed a greater capacity to inhibit oxidative hemolysis (B8, IC50 of 25 and 47.4 µg/mL for Δt of 60 and 120 min, respectively) and higher cytotoxic (B8-B13, GI50 between 7.1 and 17 µg/mL), anti-inflammatory (B13, IC50 of 10 µg/mL), and antibacterial activities. In turn, the antifungal activity varied depending on the tested fungi. All these results suggest that maturity influences the phenolic composition and bioactive properties of cardoon blades, which reveal great potential for the development of bioactive ingredients for food and pharmaceutical applications, among others.

Protective Effect of Quercetin 3-O-Glucuronide against Cisplatin Cytotoxicity in Renal Tubular Cells.

Quercetin, a flavonoid with promising therapeutic potential, has been shown to protect from cisplatin nephrotoxicity in rats following intraperitoneal injection, but its low bioavailability curtails its prospective clinical utility in oral therapy. We recently developed a micellar formulation (P-quercetin) with enhanced solubility and bioavailability, and identical nephroprotective properties. As a first aim, we herein evaluated the oral treatment with P-quercetin in rats, which displayed no nephroprotection. In order to unravel this discrepancy, quercetin and its main metabolites were measured by HPLC in the blood and urine after intraperitoneal and oral administrations. Whilst quercetin was absorbed similarly, the profile of its metabolites was different, which led us to hypothesize that nephroprotection might be exerted in vivo by a metabolic derivate. Consequently, we then aimed to evaluate the cytoprotective capacity of quercetin and its main metabolites (quercetin 3-O-glucoside, rutin, tamarixetin, isorhamnetin and quercetin 3-O-glucuronide) against cisplatin toxicity, in HK-2 and NRK-52E tubular cell lines. Cells were incubated for 6 h with quercetin, its metabolites or vehicle (pretreatment), and subsequently 18 h in cotreatment with 10-300 µM cisplatin. Immediately after treatment, cell cultures were subject to the MTT technique as an index of cytotoxicity and photographed under light microscopy for phenotypic assessment. Quercetin afforded no direct cytoprotection and quercetin-3-O-glucuronide was the only metabolite partially preventing the effect of cisplatin in cultured tubule cells. Our results identify a metabolic derivative of quercetin contributing to its nephroprotection and prompt to further explore exogenous quercetin-3-O-glucuronide in the prophylaxis of tubular nephrotoxicity.

Disclosing the Antioxidant and Neuroprotective Activity of an Anthocyanin-Rich Extract from Sweet Cherry (Prunus avium L.) Using In Vitro and In Vivo Models.

In this study, an autochthonous variety of sweet cherry (Prunus avium L.), namely "Moretta di Vignola", was processed to prepare extracts rich in polyphenols, which were characterized by high-performance liquid chromatography (HPLC) separation coupled to UV/DAD and ESI-MSn analysis. Then, a sweet cherry anthocyanin-rich extract (ACE) was prepared, fully characterized and tested for its activity against Parkinson's disease (PD) in cellular (BV2 microglia and SH-SY5Y neuroblastoma) and in Drosophila melanogaster rotenone (ROT)-induced model. The extract was also evaluated for its antioxidant activity on Caenorhabditis elegans by assessing nematode resistance to thermal stress. In both cell lines, ACE reduced ROT-induced cell death and it decreased, alone, cellular reactive oxygen species (ROS) content while reinstating control-like ROS values after ROT-induced ROS rise, albeit at different concentrations of both compounds. Moreover, ACE mitigated SH-SY5Y cell cytotoxicity in a non-contact co-culture assay with cell-free supernatants from ROT-treated BV-2 cells. ACE, at 50 µg/mL, ameliorated ROT (250 µM)-provoked spontaneous (24 h duration) and induced (after 3 and 7 days) locomotor activity impairment in D. melanogaster and it also increased survival and counteracted the decrease in fly lifespan registered after exposure to the ROT. Moreover, heads from flies treated with ACE showed a non-significant decrease in ROS levels, while those exposed to ROT markedly increased ROS levels if compared to controls. ACE + ROT significantly placed the ROS content to intermediate values between those of controls and ROT alone. Finally, ACE at 25 µg/mL produced a significant increase in the survival rate of nematodes submitted to thermal stress (35 °C, 6-8 h), at the 2nd and 9th day of adulthood. All in all, ACE from Moretta cherries can be an attractive candidate to formulate a nutraceutical product to be used for the prevention of oxidative stress-induced disorders and related neurodegenerative diseases.

Applications of bioactive compounds extracted from olive industry wastes: A review.

The wastes generated during the olive oil extraction process, even if presenting a negative impact for the environment, contain several bioactive compounds that have considerable health benefits. After suitable extraction and purification, these compounds can be used as food antioxidants or as active ingredients in nutraceutical and cosmetic products due to their interesting technological and pharmaceutical properties. The aim of this review, after presenting general applications of the different types of wastes generated from this industry, is to focus on the olive pomace produced by the two-phase system and to explore the challenging applications of the main individual compounds present in this waste. Hydroxytyrosol, tyrosol, oleuropein, oleuropein aglycone, and verbascoside are the most abundant bioactive compounds present in olive pomace. Besides their antioxidant activity, these compounds also demonstrated other biological properties such as antimicrobial, anticancer, or anti-inflammatory, thus being used in formulations to produce pharmaceutical and cosmetic products or in the fortification of food. Nevertheless, it is mandatory to involve both industries and researchers to create strategies to valorize these byproducts while maintaining environmental sustainability.

Chemical composition and biological activity of cardoon (Cynara cardunculus L. var. altilis) seeds harvested at different maturity stages.

Cardoon seeds collected in Greece at four different maturity stages (samples S1 to S4) were analysed in terms of chemical composition and in vitro bioactivities. The content of phenolic compounds (six compounds in total) increased with increasing maturity, and 3,5-O-dicaffeyolquinic (14.8-33.8 mg/g extract) acid was the compound detected in higher abundance. Mature seeds (sample S4) also revealed the highest content in lipids (23 g/100 g extract) and tocopherols (29.62 mg/100 g dw) and demonstrated the highest cytotoxic (GI50 of 97-216 µg/mL) and anti-inflammatory (IC50 = 148 µg/mL) activities, and capacity to inhibit the formation of thiobarbituric acid reactive substances (TBARS) (IC50 = 5 µg/mL). Cardoon seed hydroethanolic extracts also revealed high antibacterial and antifungal potential, particularly samples S3 and S1, respectively. This study proved the multifaceted potential associated with valorisation of cardoon seeds, while their biological and chemical composition can be influenced by the maturity stage.

Phenolic Composition and Biological Properties of Cynara cardunculus L. var. altilis Petioles: Influence of the Maturity Stage.

Hydroethanolic extracts of cardoon petioles collected at sixteen growth stages (P1-P16) were characterized in terms of their phenolic composition and bioactive potential (antioxidant, cytotoxic, anti-inflammatory, and antimicrobial activities). Fifteen phenolic compounds were tentatively identified (i.e., ten phenolic acids and five flavonoid glycosides); the main compounds were 5-O-caffeoylquinic and 1,5-di-O-caffeoylquinic acids. Samples collected at early maturity (P1-P4) presented a weak positive correlation between the higher content in polyphenols (P3: 101-mg/g extract) and better inhibition capacity against thiobarbituric acid reactive substance formation (TBARS; P3: IC50 = 5.0 µg/mL). Samples at intermediate maturation stages (P9) presented higher cytotoxic and anti-inflammatory potential. Moreover, immature petioles showed greater antihemolytic (OxHLIA; P4: IC50 = 65 and 180 µg/mL for Δt of 60 and 120 min, respectively) and antibacterial activity. The antifungal activity varied depending on the maturation stage and the fungi strain. In conclusion, the maturation stage may greatly affect the polyphenols composition and content and the bioactive potential of cardoon petioles.

Wine, Polyphenols, and Mediterranean Diets. What Else Is There to Say?

A considerable amount of literature has been published claiming the cardiovascular benefits of moderate (red) wine drinking, which has been considered a distinguishing trait of the Mediterranean diet. Indeed, red wine contains relevant amounts of polyphenols, for which evidence of their biological activity and positive health effects are abundant; however, it is also well-known that alcohol, even at a low level of intake, may have severe consequences for health. Among others, it is directly related to a number of non-communicable diseases, like liver cirrhosis or diverse types of cancer. The IARC classifies alcohol as a Group 1 carcinogen, causally associated with the development of cancers of the upper digestive tract and liver, and, with sufficient evidence, can be positively associated with colorectum and female breast cancer. In these circumstances, it is tricky, if not irresponsible, to spread any message on the benefits of moderate wine drinking, about which no actual consensus exists. It should be further considered that other hallmarks of the Mediterranean diet are the richness in virgin olive oil, fruits, grains, and vegetables, which are also good sources of polyphenols and other phytochemicals, and lack the risks of wine. All of these aspects are reviewed in this article.

Polyphenols and Human Beings: From Epidemiology to Molecular Targets.

Dietary polyphenols have been associated with health benefits in the prevention of a range of degenerative and age-related diseases that constitute the major causes of death and incapacitation in developed countries [...].