Challenges for research on polyphenols from foods in Alzheimer's disease: bioavailability, metabolism, and cellular and molecular mechanisms

Polyphenols are the most abundant antioxidants in diet. Indeed, fruits, vegetables, beverages (tea, wine, juices), plants, and some herbs are loaded with powerful antioxidant polyphenols. Despite their wide distribution, research on human health benefits truly began in the mid-1990s (Scalbert, A.; Johnson, I. T.; Saltmarsh, M. Am. J. Clin. Nutr. 2005, 81, S15S-217S). Phenolic compounds have been receiving increasing interest from consumers and manufacturers because numerous epidemiological studies have suggested associations between consumption of polyphenol-rich foods or beverages and the prevention of certain chronic diseases such as cancers and cardiovascular diseases (Manach, C.; Mazur, A.; Scalbert, A. Curr. Opin. Lipidol. 2005, 16, 77-84; Duthie, S. J. Mol. Nutr. Food Res. 2007, 51, 665-674). Furthermore, in the past 10 years, research on the neuroprotective effects of dietary polyphenols has developed considerably. These compounds are able to protect neuronal cells in various in vivo and in vitro models through different intracellular targets (Ramassamy, C. Eur. J. Pharmacol. 2006, 545, 51-64). However, it is not at all clear whether these compounds reach the brain in sufficient concentrations and in a biologically active form to exert beneficial effects. On the other hand, it has become clear that the mechanisms of action of these polyphenols go beyond their antioxidant activity and the attenuation of oxidative stress. Therefore, there is a need for more research on their intracellular and molecular targets as special pathways underlying distinct polyphenol-induced neuroprotection. The focus of this review is aimed at presenting the role of some polyphenols from fruits, vegetables, and beverages in neuroprotection and particularly in Alzheimer's disease and the research challenges in this area.     

Antibacterial activity of red and white wine against oral streptococci

Wine contains a number of biologically active compounds with beneficial effects on human health. The antibacterial action of commercial red and white wines against oral streptococci responsible for caries development and against S. pyogenes responsible for pharyngitis was studied. Its postcontact effect against S. mutans was also studied. Both wines displayed activity. The compounds responsible for such activities were succinic, malic, lactic, tartaric, citric, and acetic acid. The synthetic mixtures of the organic acids tested at the concentrations found in wine had greater antibacterial activity than the beverages, indicating that in wine they are inhibited by other components. Wine polyphenols displayed no activity against oral streptococci or S. pyogenes. Findings show that wine is active against oral streptococci and S. pyogenes and suggest that it enhances oral health.     

Guava fruit (Psidium guajava L.) as a new source of antioxidant dietary fiber.

Guava (Psidium guajava L.) is a tropical fruit, widely consumed fresh and also processed (beverages, syrup, ice cream, and jams). Pulp and peel fractions were tested, and both showed high content of dietary fiber (48.55-49.42%) and extractable polyphenols (2.62-7.79%). The antioxidant activity of polyphenol compounds was studied, using three complementary methods: (i) free radical DPPH* scavenging, (ii) ferric reducing antioxidant power assay (FRAP), and (iii) inhibition of copper-catalyzed in vitro human low-density lipoprotein (LDL) oxidation. All fractions tested showed a remarkable antioxidant capacity, and this activity was correlated with the corresponding total phenolic content. A 1-g (dry matter) portion of peel contained DPPH* activity, FRAP activity, and inhibition of copper-induced in vitro LDL oxidation, equivalent to 43 mg, 116 mg, and 176 mg of Trolox, respectively. These results indicate that guava could be a suitable source of natural antioxidants. Peel and pulp could also be used to obtain antioxidant dietary fiber (AODF), a new item which combines in a single natural product the properties of dietary fiber and antioxidant compounds.     

Effect of some phenolic compounds and beverages on pepsin activity during simulated gastric digestion

The effect of some polyphenols (resveratrol, catechin, epigallocatechin-3-gallate, and quercetin) and beverages (red wine and green tea) on the enzymatic activity of pepsin during the digestion of three different substrates (pork meat, insoluble azocasein, and denatured hemoglobin) has been investigated. The tested polyphenols and beverages increase the initial velocity of the reaction, and the activating effect is concentration dependent. The order of effectiveness of polyphenols in increasing the initial velocity of the reaction is resveratrol > or = quercetin > epigallocatechin-3-gallate > catechin. The kinetic data obtained with soluble denatured hemoglobin show that the K(m) for the substrate is not changed, whereas the V(max) of the reaction is increased. Pepsin activity follows a simple Michaelis-Menten kinetic suggesting that k(3) is increased by polyphenols. To the authors' knowledge, the present study is the first demonstration that some polyphenols and related beverages are able to enhance the enzymatic activity of pepsin.     

Identification of the strong vasorelaxing substance scirpusin B, a dimer of piceatannol, from passion fruit (Passiflora edulis) seeds

Piceatannol is present in passion fruit (Passiflora edulis) seeds in high amounts. In this study, we isolated the second major polyphenolic compound of passion fruit seeds and identified it as scirpusin B, which is a dimer of piceatannol. We investigated the antioxidant activities and vasorelaxing effects of these polyphenols. Their antioxidant effects were measured using an in vitro 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, and their vasorelaxant effects were determined ex vivo in rat thoracic aorta. Both polyphenolic compounds exhibited potent antioxidant activities and significant vasorelaxant effects in endothelium-intact aortas. More specifically, scirpusin B exerted a greater antioxidant activity and vasorelaxant effect compared with that of piceatannol. Additionally, the vasorelaxation effects of the compounds were induced via the NO derived from the endothelium. This study provides the possibility that polyphenols in passion fruit seeds are effective against cardiovascular diseases (CVDs).     

Oxidative deamination of benzylamine and lysine residue in bovine serum albumin by green tea, black tea, and coffee

Oxidative deamination by various polyphenolic compounds is presumed to be due to the oxidative conversion of polyphenols to the corresponding quinones through autoxidation. Here we examined the oxidative deamination by the polyphenol-rich beverages green tea, black tea, and coffee at a physiological pH and temperature. Green tea, black tea, and coffee extracts oxidatively deaminated benzylamine and the lysine residues of bovine serum albumin to benzaldehyde and alpha-aminoadipic delta-semialdehyde residues, respectively, in sodium phosphate buffer (pH 7.4) at 37 degrees C in both the presence and absence of Cu2+, indicating the occurrence of an amine (lysyl) oxidase-like reaction. We also examined the effects of pH and metal ions on the reaction. The possible biological effects of drinking polyphenol-rich beverages on human are also discussed.     

Effects of natural phenolic compounds on the antioxidant activity of lactoferrin in liposomes and oil-in-water emulsions

The effect of natural phenolic compounds on the antioxidant and prooxidant activity of lactoferrin was studied in liposomes and oil-in-water emulsions containing iron. The antioxidants tested with lactoferrin were alpha-tocopherol, ferulic acid, coumaric acid, tyrosol, and natural phenolic extracts obtained from three different extra-virgin olive oils and olive mill wastewater. The natural extracts of olive oils and mill wastewaters were composed mainly of polyphenols and simple phenolics, respectively. Lipid oxidation at 30 degrees C was determined by the formation of hydroperoxides and fluorescent compounds resulting from oxidized lipid interactions. All phenolic compounds showed synergistic properties in reinforcing the antioxidant activity of lactoferrin in lipid systems containing iron. The highest synergistic effects were observed for the phenolic extracts rich in polyphenols of extra-virgin olive oils and lactoferrin. This synergistic effect was higher in liposomes than in emulsions.     

Grape seed polyphenols protect cardiac cells from apoptosis via induction of endogenous antioxidant enzymes

Grape seed extract (GSE) has been reported to exert protective effects on various forms of cardiac disorders. The cardiovascular protective effects of GSE are believed to be ascribed to its antioxidative properties. A series of studies have demonstrated that polyphenols are instrumental for the antioxidative properties of GSE. This study was undertaken to investigate whether two major polyphenols isolated from GSE (catechin and proanthocyanidin B4) could increase the endogenous antioxidant enzymes in cardiomyocytes, and whether such increased cellular defenses could provide protection against oxidative cardiac cell injury. Incubation of cardiac H9C2 cells with micromolar concentrations of catechin or proanthocyanidin B4 resulted in a significant induction of cellular antioxidant enzymes in a concentration-dependent fashion. Furthermore, catechine or proanthocyanidin B4 pretreatment led to a marked reduction in xanthine oxidase (XO)/xanthine-induced intracellular reactive oxygen species (ROS) accumulation and cardiac cell apoptosis. These results indicated that grape seed polyphenols (GSP) could protect against cardiac cell apoptosis via the induction of endogenous antioxidant enzymes. This may be an important mechanism underlying the protective effects of GSE observed with various forms of cardiovascular disorders.     

Inhibition of gastrointestinal lipolysis by green tea, coffee, and gomchui (Ligularia fischeri) tea polyphenols during simulated digestion

Green tea, coffee, and gomchui (Ligularia fischeri) tea, which are rich in polyphenols, may exhibit antiobesity effects by inhibiting pancreatic lipase. However, the bioavailability of some polyphenols is poor due to either degradation or absorption difficulties in the gastrointestinal tract, thus making their beneficial effects doubtful. This study was conducted to evaluate the inhibitory effect of three beverages on lipolysis and the contribution of their major polyphenols during simulated digestion. During simulated digestion, gomchui tea was the most potent at inhibiting gastrointestinal lipolysis, whereas green tea was the least potent. The strongest lipase inhibitor among purified major polyphenols was a green tea polyphenol, (-)-epigallocatechin gallate (EGCG, IC(50) = 1.8 ± 0.57 μM), followed by di-O-caffeoylquinic acid isomers (DCQA, IC(50) from 12.7 ± 4.5 to 40.4 ± 2.3 μM), which are gomchui tea polyphenols. However, the stability of DCQA was greater than that of EGCG when subjected to simulated digestion. Taken together, gomchui tea, which has DCQA as the major polyphenol, showed stronger lipolysis inhibitory activity during simulated digestion compared to both green tea and coffee.     

How to standardize the multiplicity of methods to evaluate natural antioxidants

A great multiplicity of methods has been used to evaluate the activity of natural antioxidants by using different techniques of inducing and catalyzing oxidation and measuring the end point of oxidation for foods and biological systems. Antioxidant in vitro protocols for foods should be based on analyses at relatively low levels of oxidation under mild conditions and on the formation and decomposition of hydroperoxides. For antioxidant in vivo protocols, widely different methods have been used to test the biological protective activity of phenolic compounds. Unfortunately, many of these protocols have been based on questionable methodology to accurately measure oxidative damage and to assess relevant changes in biological targets. Many studies testing the ex vivo activity of phenolic compounds to inhibit human low-density lilpoprotein (LDL) oxidation have been difficult to evaluate because of the structural complexity of LDL particles and because a multitude of markers of oxidative damage have been used. Although studies with animal models of atherosclerosis have demonstrated the antioxidant effect of phenolic compounds in delaying the progress of this disease, human clinical trials of antioxidants have reported inconsistent and mixed results. Complex mixtures of plant polyphenols have been shown to be absorbed to varying degrees as metabolites in the intestine, but little is known about their interactions, bioavailability, and their in vivo antioxidant activity. Several metabolites identified in human plasma after consuming flavonoids need to be tested for possible nonantioxidant activities. More research and better-designed human studies are required to clarify the complex questions of bioavailability of polyphenols and the factors affecting their in vivo activities. Until we know what relevant in vivo activities to measure, any claims on the biological and health protective effects of natural polyphenolic compounds in our diet are premature.     

Inhibition of Df-protease associated with allergic diseases by polyphenol.

It was reported that Df-protease from house dust mite (Dermatophagoides farinae) catalyzes the activation of the kallikrein-kinin system in human plasma and is closely associated with mite-induced allergy. Therefore, to prevent the release of kinin by Df-protease, the inhibitory activity of polyphenols including catechins and flavonols was tested in vitro and in vivo. Among them, myricetin and epigallocatechin gallate (EGCg) effectively inhibited the amidase activity of Df-protease with K(i) values of 1 x 10(-)(8) and 6 x 10(-)(4) M, respectively. The kinin release in human plasma was extensively inhibited by the addition of EGCg in comparison with myricetin. Enhancement of vascular permeability in guinea pigs caused by Df-protease was markedly suppressed by EGCg.     

Effects of protein-polyphenol interactions on beverage haze, stabilization, and analysis

The haze-forming activity of a polypeptide depends greatly on its proline content. Haze-forming polyphenols have at least two binding groups, each of which has at least two hydroxy groups on an aromatic ring. The protein/polyphenol ratio has a strong influence on the amount of haze formed; the largest amount occurs when the numbers of polyphenol binding ends and protein binding sites are nearly equal. This has important consequences for turbidimetric methods used to measure haze-active proteins and polyphenols in beverages. The ratio also influences the effectiveness of a number of stabilization procedures.     

Isolation and tyrosinase inhibitory effects of polyphenols from the leaves of persimmon, Diospyros kaki

The main polyphenols were isolated from the leaves of six selected persimmon cultivars. Seven compounds were obtained by reverse-phase HPLC, and their structures were elucidated by multiple NMR measurements. These compounds are hyperoside, isoquercitrin, trifolin, astragalin, chrysontemin, quercetin-3-O-(2'-O-galloyl-β-D-glucopyranoside) (QOG), and kaempferol-3-O-(2'-O-galloyl-β-D-glucopyranoside) (KOG). Their inhibitory activity was tested against tyrosinase for the oxidation of L-DOPA, and only chrysontemin showed inhibitory activity. To investigate the differences of their inhibitory effects, the tyrosinase inhibitory activities of their aglycons, cyanidin, quercetin, and kaempferol, were also tested. As a result, it was confirmed that the most influential moiety for tyrosinase inhibition was the 3',4'-dihydroxy groups of the catechol moiety. Moreover, the tyrosinase inhibitory activity of chrysontemin, which was identified in persimmon leaves for the first time, is supported by a simulated model of chrysontemin docking into mushroom tyrosinase.     

Study of anticancer activities of muscadine grape phenolics in vitro

Muscadine grapes have unique aroma and flavor characteristics. Although a few studies reported high polyphenols content of muscadine grapes, little research has been conducted to evaluate the phenolic compounds bioactivities in any muscadine grape cultivar. The objective of this study was to evaluate the effect of phenolic compounds in muscadine grapes on cancer cell viability and apoptosis. Four cultivars of muscadine (Carlos, Ison, Noble, and Supreme) were assessed in this study. Phenolic compounds were extracted from muscadine skins and further separated into phenolic acids, tannins, flavonols, and anthocyanins using HLB cartridge and LH20 column. Some individual phenolic acids and flavonoids were identified by HPLC. Anthocyanin fractions were more than 90% pure. The effect of different fractions on the viability and apoptosis of two colon cancer cell lines (HT-29 and Caco-2) was evaluated. A 50% inhibition of cancer cell population growth for the two cell lines was observed at concentrations of 1-7 mg/mL for crude extracts. The phenolic acid fractions showed a 50% inhibition at the level of 0.5-3 mg/mL. The greatest inhibitory activity was found in the anthocyanin fraction, with a 50% inhibition at concentrations of approximately 200 microg/mL in HT-29 and 100-300 microg/mL in Caco-2. Anthocyanin fractions also resulted in 2-4 times increase in DNA fragmentation, indicating the induction of apoptosis. These findings suggest that polyphenols from muscadine grapes may have anticancer properties.     

Control of MicroRNA Expression as a New Way for Resveratrol To Deliver Its Beneficial Effects

Grapes produce large amounts of polyphenols. Many of them accumulate in the skin, pulp, and seeds and are consequently found in wine. The health benefits of a moderate consumption of wine have been attributed at least in part to grape's polyphenols. Among them, resveratrol (3,5,4'-trihydroxystilbene) is a phytoalexin that stimulates plant cell defenses against infections and also plays protective roles in humans, where it delays cardiovascular alterations and exerts anticancer and anti-inflammatory effects. Despite numerous studies, the molecular mechanisms of resveratrol action are only partially understood. Given its pleiotropic effects, it was previously suggested that resveratrol protective properties may arise from its modulation of the expression of microRNAs. Therefore, this review will focus on the effects of resveratrol on microRNA populations in humans and human cell lines, especially emphasizing the microRNAs that have been implicated in resveratrol effects on inflammation, cancer, metabolism, and muscle differentiation.     

Antioxidant Activities of Extracts from Teas Prepared from Medicinal Plants, Morus alba L., Camellia sinensis L., and Cudrania tricuspidata , and Their Volatile Components

The antioxidant activity of essences of teas prepared from mulberry ( Morus alba L.), Camellia sinensis L., and Cudrania tricuspidata (Carr.) Burea plant was examined using two antioxidant assays. Selected volatile chemicals identified in these plants were also tested for antioxidant activity. All extracts exhibited antioxidant activity with a clear dose response in the aldehyde/carboxylic acid and the malonaldehyde/gas chromatography (MA/GC) assays. Antioxidant activity of extracts at the level of 500 μg/mL ranged from 77.02 ± 0.51% (stems of Burea plant) to 52.57 ± 0.92% (fermented tea of Camellia and stems of Mulberry tea) in the aldehydes/carboxylic acid assay. Their antioxidant activity at the level of 160 μg/mL ranged from 76.17 ± 0.27% (roots of Burea plant) to 59.32 ± 0.27% (stems of Mulberry tea) in the MA/GC assay. Among the positively identified compounds (11 terpenes and terpenoids, 15 alkyl compounds, 26 nitrogen containing heterocyclic compounds, 9 oxygen containing heterocyclic compounds, 18 aromatic compounds, 7 lactones, 6 acids, and 4 miscellaneous compounds), eugenol, 2,5-dihydroxyl acetophenone, and isoeugenol exhibited antioxidant activity comparable to that of BHT in both assays. Vanillin and 2-acetylpyrrole showed potent antioxidant activity in the aldehydes/carboxylic acid assay but only moderate activity in the MA/GC assay. These results suggest that consumption of antioxidant-rich beverages prepared from these plants may be beneficial to human health.     

Potential food additives from Carex distachya roots: identification and in vitro antioxidant properties

In the present study, the chemical composition and antioxidant properties of root methanol extract of Carex distachya Desf. (Cyperaceae) were assessed to use this plant as sources of food additives and nutraceuticals. The IC50 of the extract (4.2 microg/mL), derived from the DPPH radical scavenging capacity assay, was similar to those of ascorbic acid, alpha-tocopherol, and BHT. These results revealed a strong antioxidant activity because of the presence of an extraordinary quantity of bioactive phytochemicals. The phytochemical study of the root extract led to the isolation and identification of new and known polyphenols, most of them common constituents of plant foods. A total of 16 polyphenols, identified on the basis of spectroscopic data as 7 lignans, 4 phenylethanoids, 3 resveratrol derivatives, a monolignol, and a secoiridoid glucoside, were isolated. The tentative structural elucidation of the new metabolites 5'-O-beta-D-glucopyranosyloxy-3,3'-dimethoxy-7,9'-epoxylignan-4,8',9-triol and 3,5-bis-O-beta-D-glucopyranosyloxy-3'-methoxy-trans-stilben-4'-ol have been performed by a combined approach using ESI/TQ/MS techniques and 1D and 2D NMR experiments. All of the compounds have been tested for their antioxidant activity using six different antioxidant and radical scavenging tests. Interestingly, the extract contained high quantities of polyphenols, most of them reported as constituents of edible plants, such as grape and olive, suggesting that the methanol root extract of this plant could be used as a source of natural antioxidants useful as potential food additives.     

Effects of tea polyphenols on the invasion and matrix metalloproteinases activities of human fibrosarcoma HT1080 cells

The effects of tea polyphenols on the invasion of highly metastatic human fibrosarcoma HT1080 cells through a monolayer of human umbilical vein endothelial cells (HUVECs) and the accompanying basal membrane were investigated. Among the tea polyphenols tested, epicatechin gallate (ECg), epigallocatechin gallate (EGCg), and theaflavin strongly suppressed the invasion of HT1080 cells into the monolayer of HUVECs/gelatin membrane, whereas epicatechin, epigallocatechin, tea flavonols, tea flavones, and gallate derivatives had no effect. Both theaflavin-digallate and theasinensin D showed a weak invasion inhibitory effect. ECg significantly inhibited the invasion without cytotoxicity against cancer cells and HUVECs. Ester-type catechins (ECg and EGCg) and theaflavin strongly suppressed the gelatin degradation mediated by matrix metalloproteinase (MMP) 2 and MMP-9, which were secreted into the conditioned medium of HT1080 cells. In conclusion, among the tea polyphenols tested, ECg was considered to be the agent with the most potential antimetastasis activity because it inhibited invasion in the absence of cytotoxicity.     

The role of iron and the factors affecting off-color development of polyphenols

Iron deficiency affects over two billion people worldwide (Lotfi, M.; Venkatesh Mannar, M. G.; Merx, R. J.; Naber-van den Heuvel, P. Micronutrient Fortification of Foods: Current Practices, Research,and Opportunities; Micronutrient Initiative: Ottawa, Ontario, Canada, 1996). However, fortifying foods with highly bioavailable iron is technically challenging because of off-color and off-flavor development, catalytic degradation of vitamins, and oxidation of lipids. The role of highly bioavailable iron in the off-color development of foods and beverages is not well-understood. The goal of this research was to examine the interaction of iron with simple phenolics and polyphenols. Factors that may affect off-color development, such as pH, oxygen, temperature, and reducing and chelating agents, were evaluated as a model for food products. Our results demonstrated that the iron that reacts with the simple phenolic, catechol, to develop off-color must be in the oxidized state, and the iron is reduced in the presence of catechol. Because this is an oxidation/reduction reaction, the redox potential of all of the components is critical to the color development. Ferrous iron sources with low redox potentials and ferric iron sources with high redox potentials caused off-color development with catechol. Only polyphenols that contain ortho-hydroxyl groups cause off-color development with iron. All of the factors tested affect off-color development and redox potential of the system. Low pH, low oxygen, high temperature, and the presence of reducing and chelating agents inhibited off-color development. To confirm the model, foods that contained these polyphenols were evaluated for off-color development when iron was added. The foods tested reacted similarly to the models of polyphenols with iron. Off-color development was caused by oxidation-reduction interactions between ferric iron and polyphenols that contained ortho-dihydroxyl groups. Ferrous iron needed to be oxidized to participate in off-color development. In addition, methods identified in the models to prevent off-color development were effective in most of the food products examined. Using the ferrous form of iron and maintaining it in its reduced form by lowering pH, removing oxygen, and including reducing agents, it was possible to fortify foods with highly bioavailable iron.     

Polyphenols and health: current state and progress

During the 5th International Conference on Polyphenols and Health that was held in Sitges (Spain) in October 2011, the latest advances in this area of active research were presented. Sessions on polyphenol effects on cardiovascular disease, polyphenols as ingredients of functional foods, the role of polyphenols in preventing obesity and diabetes, the interaction of polyphenols with gut microbiota, bioavailability and metabolism of polyphenols in humans, the mechanisms of action of these metabolites in different models, new methodologies for the study of the role of polyphenols in health, polyphenols and cancer, recent developments in phenolic compounds and neuroscience, and polyphenols in epidemiology and public health were organized. This highlight issue presents a selection of papers from invited speakers, oral presentations, and poster prize winners. The perspectives for this exciting area of very active research were also discussed at the meeting and are summarized in this introductory paper.