Chemical Characterization and Biological Properties of Leguminous Honey.

Honey can beneficially act against different human diseases, helping our body to improve its health. The aim of the present study was first to increase knowledge of some biochemical characteristics (amount and composition of polyphenols and volatile organic compounds, vitamin C content) of five Italian legume honeys (alfalfa, astragalus, carob, indigo, and sainfoin). Furthermore, we evaluated their potential health properties by studying their antioxidant and in vitro anti-inflammatory activities and in vitro inhibitory effects on three enzymes involved in neurodegenerative diseases (acetylcholinesterase, butyrylcholinesterase, and tyrosinase). Alfalfa honey showed the highest total polyphenol content (TPC) (408 µg g-1 of product). Indigo honey showed the lowest TPC (110 µg g-1 of product). The antioxidant activity was noteworthy, especially in the case of sainfoin honey (IC50 = 6.08 mg), which also exhibited excellent inhibitory action against butyrylcholinesterase (74%). Finally, the correlation between the biochemical and functional results allowed us to identify classes of molecules, or even single molecules, present in these five honeys, which are capable of influencing the properties indicated above.

In vitro prospective healthy and nutritional benefits of different Citrus monofloral honeys.

We studied the total polyphenols, flavonoids, vitamin C, the antioxidant and anti-inflammatory activity of six Citrus monofloral honey, and the in vitro inhibitory effect against cholinesterases and tyrosinase. Finally, we assessed their effect against the biofilm of some pathogenic bacteria. Lime honey showed the best antioxidant activity and the highest content of polyphenols and vitamin C. Lemon and tangerine honey contained almost exclusively flavonoids. Lemon honey better preserved the bovine serum albumin against denaturation (IC50 = 48.47 mg). Honeys inhibited acetylcholinesterase, butyrylcholinesterase, and tyrosinase up to 12.04% (tangerine), 19.11% (bergamot), and 94.1% (lemon), respectively. Lime and clementine honey better inhibited the Listeria monocytogenes biofilm. Bergamot honey acted mainly against the Staphylococcus aureus and Acinetobacter baumannii biofilm; bergamot and tangerine honey inhibited the Pseudomonas aeruginosa biofilm particularly. Bergamot, clementine, and tangerine honey acted against Escherichia coli sessile cell metabolism. This Citrus honey exhibited in vitro prospective health benefits and is applicable for future in vivo studies.

Enriched pasta incorporating typical vegetables of mediterranean diet: in vitro evaluation of inhibitory potential on digestive enzymes and predicted glycaemic index.

Swiss chard (Beta vulgaris L.) and chicory (Cichorium intybus L.) contain biologically active compounds with proven health benefits. Durum wheat noodle-shaped pasta enriched with dried and powder leaves of chard or chicory, at two different levels of supplementation (3%, 6%) was prepared on a laboratory scale. The content of polyphenols, pigments, carotenoids, in vitro inhibition of digestive enzymes and the predicted glycaemic response of the fortified pasta were evaluated. All formulations showed in vitro enzyme inhibition of amylase, glucosidase, and lipase and a low pGI <43. The lowest predicted glycaemic index (pGI = 34 ± 1.1) was found for pasta enriched with 3% beet powder. The incorporation of Beta vulgaris and Cichorium intybus leaf powders improved the nutritional properties of the pasta and also imparted an attractive natural colour to the products.

Biochemical Characterization of Some Varieties of Apricot Present in the Vesuvius Area, Southern Italy.

The witnesses of the millenary history of Campania felix in southern Italy highlighted that several fruit and vegetables cultivated in such territory could potentially be a treasure trove of important health elements. Our work evaluated the content of ß-carotene, ascorbic acid, and total phenolics and the antioxidant activity of ten typical varieties of apricots cultivated in the Vesuvius area in the Campania region. The total polyphenols varied between 10.24 and 34.04 mg/100 g of a fresh sample. The amount of ascorbic acid also varied greatly, ranging from 2.65 to 10.65 mg/100 g of a fresh product. B-Carotene reached values up to 0.522 mg/100 g of the fresh sample. The correlation analysis performed, accounting for these parameters, showed that the antioxidant activity, calculated by 2,2-diphenyl-1-picrylhydrazyl (DPPH assay) and azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) tests, was influenced mainly by the content of total polyphenols, with ρ = -0.762 and ρ = 0.875 when we considered DPPH and ABTS tests, respectively, slightly less by the content of ascorbic acid, and not by ß-carotene. The dendrogram clustered eight varieties into two main groups; on the other hand, two varieties ("Vitillo" and "Preveta bella") seemed hierarchically distant. The gas chromatography-mass spectrometry (GC-MS) analysis of volatile organic compounds (VOCs), herein performed for the first time, demonstrated the influence of the varieties on the VOC profiles, both from a qualitative and semiquantitative perspective, discriminating the varieties in different clusters, each of which was characterized by specific notes. α-Terpinolene was the only terpene identified by GC-MS that appeared to affect the antioxidant activity.

Lowering the predicted glycemic index of pasta using dried onions as functional ingredients.

Pasta is a commonly consumed food; adding some ingredients, maybe turn it into a functional food with health benefits. These ingredients consist of dietary fiber, antioxidant molecules, and enzyme inhibitor compounds, related to a reduced risk for some diseases. Onion (Allium cepa L.) is a rich source of bioactive compounds rendering it a relevant candidate for the production of functional foods. The present study examines the in vitro starch digestibility of durum wheat pasta supplemented with 3% onion flour. The incorporation of onion flour attenuated the extent of starch digestion and accordingly the area under the curve of reducing sugars discharged during in vitro digestion. The predicted glycemic index (pGI) of pasta supplemented with onion flour (3%) was significantly lower (pGI = 54 ± 0.17) than the control pasta (pGI = 72 ± 0.14). These results indicate that 3% onion fortified pasta represents a food with potential healthy properties, showing glucose-lowering capabilities in vitro.

Fatty Acid Composition, Antioxidant, and in vitro Anti-inflammatory Activity of Five Cold-Pressed Prunus Seed Oils, and Their Anti-biofilm Effect Against Pathogenic Bacteria.

Background/Aim: Sweet almond (Prunus amygdalus dulcis) oil is one of the most famous cold-pressed seed oils. However, other species of Prunus can provide oils with healthy properties. We analyzed the fatty acid (FA) composition, as well as the antioxidant, the in vitro anti-inflammatory properties, and the antibiofilm activity of five commercial vegetable cold-pressed seed oils of apricot, peach, plum, cherry, and black cherry. Methods: Gas Chromatography-Mass Spectrometry was performed for the analysis of FAs The antioxidant property of the oils was carried using different tests [2, 2-diphenyl-1-picrylhydrazyl (DPPH assay)], Ferric Reducing Antioxidant Power (FRAP), and the 2, 20 -azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS·+). The denaturation assay performed on bovine serum albumin (BSA) was used to evaluate the in vitro anti-inflammatory activity. The anti-biofilm activity was assessed using five pathogenic strains, namely, Acinetobacter baumannii, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus, through the crystal violet test and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), used to evaluate the metabolism of the microbial cells present within the biofilm. Results: Oleic acid and linoleic acids were the most abundant FAs. Black cherry seed oil exhibited the best antioxidant activity, but in general, the amount of oil needed to inhibit the activity of 1 ml of DPPH assay at 50% did not exceed 10 µg. The extract concentration for the 50% inhibition of the denaturation of the protein (IC50) did not exceed 4.4 µg. Linoleic and stearic acids affected the antioxidant activity of the oils; oleic acid, linolenic, and palmitoleic acids exhibited beneficial effects in preserving the BSA denaturation, as shown by the correlation data. The oils were able to inhibit the biofilm formation of the pathogens (up to 71.40% of inhibition) as well as act against their mature biofilm, although with different strengths, with values up to 61.54%. Concurrently, they also acted on the pathogen metabolism. Conclusion: The oils represent a valuable source of some healthy FAs. They showed potential antioxidant and anti-inflammatory in vitro activity, in addition, their potential effect on the biofilm can offer important ideas for research and reflection on their use as functional foods and/or ingredients.

Polyphenols Content and In Vitro α-Glycosidase Activity of Different Italian Monofloral Honeys, and Their Effect on Selected Pathogenic and Probiotic Bacteria.

We evaluated the polyphenol content and the α-glucosidase activity exhibited by different monofloral honeys of Italian origin. Their capacity to act on different pathogenic (Acinetobacter baumannii, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus) as well as probiotic bacteria (Lacticaseibacillus casei, Lactobacillus acidophilus, Lactiplantibacillus plantarum, Lactobacillus gasseri, and Lacticaseibacillus rhamnosus) was also assessed. Total polyphenols varied between 110.46 µg/g of fresh product (rhododendron honey) and 552.29 µg/g of fresh product (strawberry tree honey). Such result did not correspond to a parallel inhibitory α-glycosidase activity that, in each case was never higher than 33 µg/mL. Honeys were differently capable to fight the biofilm formation of the pathogens (inhibition up to 93.27%); they inhibited the in vitro adhesive process (inhibition up to 84.27%), and acted on mature biofilm (with values up to 76.64%). Their effect on bacterial metabolism was different too. Honeys were ineffective to inhibit E. coli mature biofilm nor to act on its metabolism. The action of the honey on probiotic strains seemed almost always stimulate their growth. Thus, these monofloral honeys might exhibit effects on human health and act positively as prebiotics.

Polyphenols, Antioxidant, Antibacterial, and Biofilm Inhibitory Activities of Peel and Pulp of Citrus medica L., Citrus bergamia, and Citrus medica cv. Salò Cultivated in Southern Italy.

The aim of this paper was to study the polyphenols of peel and pulp of three Citrus taxa-Citrus medica, Citrus bergamia, and Citrus medica cv. Salò-cultivated in the Cosenza province, Southern Italy, and to evaluate their antioxidant and antibacterial activity, performed against Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Staphylococcus aureus, and Pectobacterium carotovorum. Furthermore, we assessed the inhibitory effect of the extracts on bacterial capacity to form biofilm, and on the metabolic activity of the cells present therein. The results indicated that such extracts could find new potential applications in the field of natural antioxidant and anti-bacterial agents in pharmaceutics, agriculture, and food fields.

Dietary compounds and cutaneous malignant melanoma: recent advances from a biological perspective.

Cutaneous malignant melanoma is a heterogeneous disease, being the consequence of specific genetic alterations along several molecular pathways. Despite the increased knowledge about the biology and pathogenesis of melanoma, the incidence has grown markedly worldwide, making it extremely important to develop preventive measures. The beneficial role of correct nutrition and of some natural dietary compounds in preventing malignant melanoma has been widely demonstrated. This led to numerous studies investigating the role of several dietary attitudes, patterns, and supplements in the prevention of melanoma, and ongoing research investigates their impact in the clinical management and outcomes of patients diagnosed with the disease. This article is an overview of recent scientific advances regarding specific dietary compounds and their impact on melanoma development and treatment.

Alpha-amylase, α-glucosidase and lipase inhibiting activities of polyphenol-rich extracts from six common bean cultivars of Southern Italy, before and after cooking.

Common beans (Phaseolus vulgaris) are a good source of nutrients and phenolic compounds with versatile health benefits. Polyphenol-rich extracts of six ecotypes of P. vulgaris were analysed to determine their phenolic profiles and assayed in vitro for inhibitory effects on digestive enzymes relevant to carbohydrates and lipids metabolism. The extracts inhibited enzyme activities in a dose-dependent manner. IC 50 values ranged from 69 ± 1.9 to 126 ± 3.2 µg/mL and from 107.01 ± 4.5 to 184.20 ± 5.7 µg/mL, before and after cooking, for α-amylase, from 39.3 ± 4.4 to 74.13 ± 6.9 µg/mL and from 51 ± 7.7 to 122.1 ± 5.2 µg/mL for α-glucosidase and from 63.11 ± 7.5 to 103.2 ± 5.9 µg/mL and from 92.0 ± 6.3 to 128.5 ± 7.4 µg/mL for lipase. Results suggest encouraging their consumption, being natural sources of enzyme inhibitors important for type-2 diabetes and obesity prevention/control. Well-monitored in vivo studies would help to establish their beneficial effects, making them worthwhile of further consideration as functional foods.

Biochemical and biological characterization of two Brassicaceae after their commercial expiry date.

Two Brassicaceae (Eruca sativa, Brassica oleracea var. sabauda) were stored in air and under a modified atmosphere for several days after their expiry date and then analyzed. The polyphenol content and composition, as well as the antioxidant activity of the extracts, were assessed, compared to the fresh products. Antimicrobial properties on tester strains (Bacillus cereus, Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa) and in vitro anti-proliferative activity were evaluated. The cabbage extracts exhibited antimicrobial activity mainly on the ninth day after the expiry date and retained significant inhibitory effects against colon carcinoma (CaCo-2) cells. The rocket salad extract exhibited antiproliferative but not antimicrobial activity. Overall, our results indicated that they might represent a good source of natural antioxidants with antimicrobial and anti-proliferative activity, also after their expiry date, suggesting their exploitation for the recovery of important biomolecules used in the food and health industry.

Isoflavone Extracts Enhance the Effect of Epidermal Growth Factor Receptor Inhibitors in NSCLC Cell Lines.

AIM: We investigated the effects of the pharmacological inhibition in vitro of epidermal growth factor receptor (EGFR) in combination with isoflavones. MATERIALS AND METHODS: Four anticancer drugs (erlotinib, gefitinib, afatinib and AZD9291) were combined with soy and red clover isoflavone extracts and used in cellular proliferation assays. The antitumor activity of inhibitors alone and in combination with isoflavone extracts was compared on three non-small cell lung cancer (NSCLC) cell lines with affiant EGFR genotype: A549 (EGFR wt); H1795 (EGFR T790M); HCC827 (EGFR del E746-A750). RESULTS: Combined treatment with extracts significantly enhanced the antiproliferative activity of all inhibitors against these cell lines. Bioactive compounds of extracts may synergize the antitumor efficacy of the inhibitors. CONCLUSION: To date, as far as we are aware, this is the first report of the combined effect of isoflavone extracts and EGFR inhibitors on human NCSLC cell growth. Sequential treatment with these drugs combined with isoflavones may represent the basis for a new therapeutic approach.

Biochemical Characterization and Antimicrobial and Antifungal Activity of Two Endemic Varieties of Garlic (Allium sativum L.) of the Campania Region, Southern Italy.

Extracts of the bulbs of the two endemic varieties "Rosato" and "Caposele" of Allium sativum of the Campania region, Southern Italy, were analyzed. The phenolic content, ascorbic acid, allicin content, and in vitro antimicrobial and antifungal activity were determined. Ultra performance liquid chromatography with diode array detector performed polyphenol profile. The polyphenolic extracts showed antioxidant activity (EC50) lower than 120 mg. The amount of ascorbic acid and allicin in the two extracts was similar. Polyphenol extract exhibited antimicrobial activity against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and (only by the extract of Rosato) against Bacillus cereus. The extract of Caposele was more effective in inhibiting the growth of Aspergillus versicolor and Penicillum citrinum. On the other hand, the extract of Rosato was effective against Penicillium expansum.

Phenolic Composition and Antioxidant and Antiproliferative Activities of the Extracts of Twelve Common Bean (Phaseolus vulgaris L.) Endemic Ecotypes of Southern Italy before and after Cooking.

Beans are important dietary components with versatile health benefits. We analysed the extracts of twelve ecotypes of Phaseolus vulgaris in order to determine their phenolic profiles, antioxidant activity, and the in vitro antiproliferative activity. Ultra-performance liquid chromatography with diode array detector (UPLC-DAD) admitted us to detect and quantify some known polyphenols, such as gallic acid, chlorogenic acid, epicatechin, myricetin, formononetin, caffeic acid, and kaempferol. The antioxidant activity (AA) ranged from 1.568 ± 0.041 to 66.572 ± 3.197 mg necessary to inhibit the activity of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical by 50% (EC50). The extracts, except those obtained from the nonpigmented samples, were capable of inhibiting the proliferation of the human epithelial colorectal adenocarcinoma (Caco-2) cells, human breast cancer cells MCF-7, and A549 NSCLC cell line. Cultivars differed in composition and concentration of polyphenols including anthocyanins; cooking affected the antioxidant activity only marginally. Qualitative and quantitative differences in phenolic composition between the groups of beans influenced the biological activities; on the other hand, we did not find significant differences on the biological activities within the same variety, before and after cooking.

In vitro antioxidant, antimicrobial and anti-proliferative activities of purple potato extracts (Solanum tuberosum cv Vitelotte noire) following simulated gastro-intestinal digestion.

Analyses of antioxidant and in vitro antimicrobial and anti-proliferative activities of anthocyanin-rich extracts from purple potatoes, Solanum tuberosum L. cv Vitelotte noire (Solanaceae), were performed by simulating both a domestic cooking process and human digestion. Extracts of crude and cooked purple potato did not exhibit antimicrobial activity against the tester strains: Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa. The behaviour changed after the simulated gastrointestinal transit, when an inhibition halo was observed against all tester strains used, ranging from 0.53 cm against B. cereus to 0.82 cm against E. coli. In addition antioxidant activity exhibited, before and after the simulated gastrointestinal digestion (5.96 mg/mL ± 0.92; 28 mg/mL ± 0 .13, respectively) and the persistence of anti-proliferative activity against the colon cancer cells Caco-2, SW48 and MCF7, MDA-MB-231 breast cancer cells, after the simulated digestion, (EC50 = 0.21; 1.13 µg/mL), suggest that vitelotte consumption might bring tangible benefits for human health.

Nuclear receptor-induced transcription is driven by spatially and timely restricted waves of ROS. The role of Akt, IKKα, and DNA damage repair enzymes.

Gene expression is governed by chromatin mainly through posttranslational modifications at the N-terminal tails of nucleosomal histone proteins. According to the histone code theory, peculiar sets of such modifications (marks) give rise to reproducible final effects on transcription and, very recently, a further level of complexity has been highlighted in binary switches between specific marks at adjacent residues. In particular, disappearance of dimethyl-lysine 9 in histone H3 is faced by phosphorylation of the following serine during activation of gene expression. Demethylation of lysine 9 by the lysine-specific demethylase 1 (LSD1) is a pre-requisite for addition of the phosphoryl mark to serine 10 and an essential step in the transcriptional control by estrogens. It generates a local burst of oxygen reactive species (ROS) that induce oxidation of nearby nucleotides and recruitment of repair enzymes with a consequent formation of single or double stranded nicks on DNA that modify chromatin flexibility in order to allow correct assembly of the transcriptional machinery. We describe here the molecular mechanism by which members of the family of nuclear receptors prevent the potential damage to DNA during transcription of target genes elicited by the use of ROS to shape chromatin. The mechanism is based on the presence of phosphorylated serine 10 in histone H3 to prevent unbalanced DNA oxidation waves. We also discuss the opportunities raised by the use of voluntary derangement of this servo system to induce selective death in hormone-responsive transformed cells.

Retinoic acid impairs estrogen signaling in breast cancer cells by interfering with activation of LSD1 via PKA.

More than 70% of breast cancers in women require estrogens for cell proliferation and survival. 17ß-estradiol (E2) effect on mammary target cells is almost exclusively mediated by its binding to the estrogen receptor-α (ERα) that joins chromatin where it assembles active transcription complexes. The proliferative and pro-survival action of estrogens is antagonized in most cases by retinoic acid (RA), even though the cognate retinoic acid receptor-α (RARα) cooperates with ERα on promoters of estrogen-responsive genes. We have examined at the molecular level the crosstalk between these nuclear receptors from the point of view of their control of cell growth and show here that RA reverts estrogen-stimulated transcription of the pivotal anti-apoptotic bcl-2 gene by preventing demethylation of dimethyl lysine 9 in histone H3 (HeK9me2). As we previously reported, this is obtained by means of E2-triggered activation of the lysine-specific demethylase 1 (LSD1), an enzyme that manages chromatin plasticity in order to allow specific movements of chromosomal regions within the nucleus. We find that E2 fuels LSD1 by inducing migration of the catalytic subunit of protein kinase A (PKA) into the nucleus, where it targets estrogen-responsive loci. RA rescues LSD1-dependent disappearance of H3K9me2 at bcl-2 regulatory regions upon the prevention of PKA assembly to the same sites.

Highlighting chromosome loops in DNA-picked chromatin (DPC).

Growing evidence supports the concept that dynamic intra- and inter-chromosomal links between specific loci contribute to the creation of cell-type specific gene expression profiles. Therefore, analysis of the establishment of peculiar functional correlations between sites, also distant on linear DNA, that govern the transcriptional process appears to be of fundamental relevance. We propose here an experimental approach showing that 17ß-estradiol-induced transcription associates to formation of loops between the promoter and termination regions of hormone-responsive genes. This strategy reveals as a tool to be also suitably used, in conjunction with automated techniques, for an extensive analysis of sites shared by multiple genes for induced expression.

DNA oxidation as triggered by H3K9me2 demethylation drives estrogen-induced gene expression.

Modifications at the N-terminal tails of nucleosomal histones are required for efficient transcription in vivo. We analyzed how H3 histone methylation and demethylation control expression of estrogen-responsive genes and show that a DNA-bound estrogen receptor directs transcription by participating in bending chromatin to contact the RNA polymerase II recruited to the promoter. This process is driven by receptor-targeted demethylation of H3 lysine 9 at both enhancer and promoter sites and is achieved by activation of resident LSD1 demethylase. Localized demethylation produces hydrogen peroxide, which modifies the surrounding DNA and recruits 8-oxoguanine-DNA glycosylase 1 and topoisomeraseIIbeta, triggering chromatin and DNA conformational changes that are essential for estrogen-induced transcription. Our data show a strategy that uses controlled DNA damage and repair to guide productive transcription.

beta-Endorphin concentration in colostrums of Burkinabe and Sicilian women.

OBJECTIVE: Endorphins (EPs) present in human colostrum may be relevant for immediate postnatal fetal adaptation because this compound is involved in stress response and adaptation mechanisms. Endorphin levels in human colostrum are two-fold greater than corresponding maternal plasma levels; however, the high endorphin levels in human milk decrease as lactation continues. The aim of this study was to determine the beta-EP concentration in colostrums of women residing in Burkina Faso and Sicily. In addition, we investigated the source of potential differences in beta-EP levels between these populations, especially ethnic sources of these deviations. METHODS: The concentration of beta-EP was determined in the colostrum from the first 3 d subsequent to delivery by an enzyme immunoassay as immunoreactive material (IRM). RESULTS: The production of beta-EP in the colostrum was significantly higher in Burkinabe mothers (0.83 +/- 0.04 ng/mL) than in Sicilian mothers (0.31 +/- 0.02 ng/mL) at 24 h after delivery. Colostrum levels of beta-EP declined progressively during the first 3 d after delivery in both populations (0.64 +/- 0.1 and 0.28 +/- 0.015 ng/mL, respectively, at 72 h). The level of beta-EP-IRM correlated significantly with pain and psychological involvement during and after delivery. In addition, the correlation between beta-EP-IRM and length of stage II of labor was significant (P < 0.0001) in the colostrums of Sicilian mothers who received ergot derivatives, episiorrhaphy, and child birth preparation. The correlation between beta-EP-IRM and length of stage II was less significant (P < 0.001) in the colostrums of Burkinabe mothers who received neither ergot derivatives nor child birth preparation. CONCLUSION: During the first 3 d after labor the beta-EP-IRM concentration in the colostrums of Burkinabe mothers differs from that of Sicilians. In addition, because Burkinabe women produce a larger volume of colostrum, their newborns receive, during the first days of life, a larger absolute amount of beta-EP-IRM, likely resulting in better postnatal fetal adaptation.