Introduction: Polyphenols of wine have been extensively studied in relation to their health promoting properties. Red wine polyphenols consisted of different class of compounds belonging to flavonoid pathway, such as the resveratrol, quercetin, and anthocyanins which have long been considered to reduce the incidence of mortality and morbidity from cardiovascular diseases (CVD). Endothelial and monocyte activation is a pivotal event in atherosclerotic CVD, in this study was examined the anti-inflammatory effects of Italian red wines polyphenols in human vascular cells, such as endothelial and monocytoid cells. Since the wine polyhenols depend on different factors such as grape cultivar, and wine-making practices, aim of this study was to examine the anti-atherogenic effects of polyphenolic extracts from Italian red wines obtained by two Apulian grape cultivar: Primitivo and Negroamaro. Methods: Six Apulian red wines produced by the cv Negramaro and Primitivo grapes from the 2007-2008 vintage were analyzed. For each wine, triplicate lipophilic fraction analyses were carried out as per Giovinazzo et al., 2005. HPLC wine anthocyanin analysis was performed by direct injection of wine samples as reported by De Villiers et al. 2004. Human Umbilical Vein Endothelial Cells (HUVEC) and human monocytoid cells (U937) were pre-treated with chemically synthesised polyphenols, or with Primitivo and Negroamaro polyphenolic extracts and the corresponding de-alcoholised wines, at 0,2-2 % (v/v), before stimulation with 20 nmol/L phorbol myristate acetate (PMA) or 2 mg/mL lypopolysaccaride (LPS) for 24 h. Then, HUVEC were tested for the expression of Vascular Cell Adhesion Molecule(VCAM)-1 by enzyme immunoassays (EIA), and U937 supernatants were tested for the release of matrix metalloproteinases (MMP)-9 by gelatine zymography. Results: All chemically synthesized polyphenol tested reduced, in a concentration dependent manner, the stimulated expression of VCAM-1 in LPS triggered HUVEC. In the same conditions, lipophilic fraction extracted from both Primitivo and Negroamaro wines as well as the corresponding de-alcoholised wines exhibited an even higher inhibition of VCAM-1 expression (up to 50-70 fold higher than those expected on the base of individual polyphenol content). Similarly, either Primitivo and Negroamaro polyphenolic extracts reduced the PMA stimulated release of MMP-9 by PMA triggered U937.Conclusions: In this study different class of polyphenols from Primitivo and Negramaro red wines were characterised. These fractions, significantly inhibited the monocyte-derived extracellular matrix proteases release as well as the endothelial expression of athero-adhesion molecules. These inhibitory effects could explain, at least in part, the anti-inflammatory and cardio-protective properties of red wine polyphenols.

Introduction: Polyphenols of wine have been extensively studied in relation to their health promoting properties. Red wine polyphenols consisted of different class of compounds belonging to flavonoid pathway, such as the resveratrol, quercetin, and anthocyanins which have long been considered to reduce the incidence of mortality and morbidity from cardiovascular diseases (CVD). Endothelial and monocyte activation is a pivotal event in atherosclerotic CVD, in this study was examined the anti-inflammatory effects of Italian red wines polyphenols in human vascular cells, such as endothelial and monocytoid cells. Since the wine polyhenols depend on different factors such as grape cultivar, and wine-making practices, aim of this study was to examine the anti-atherogenic effects of polyphenolic extracts from Italian red wines obtained by two Apulian grape cultivar: Primitivo and Negroamaro. Methods: Six Apulian red wines produced by the cv Negramaro and Primitivo grapes from the 2007-2008 vintage were analyzed. For each wine, triplicate lipophilic fraction analyses were carried out as per Giovinazzo et al., 2005. HPLC wine anthocyanin analysis was performed by direct injection of wine samples as reported by De Villiers et al. 2004. Human Umbilical Vein Endothelial Cells (HUVEC) and human monocytoid cells (U937) were pre-treated with chemically synthesised polyphenols, or with Primitivo and Negroamaro polyphenolic extracts and the corresponding de-alcoholised wines, at 0,2-2 % (v/v), before stimulation with 20 nmol/L phorbol myristate acetate (PMA) or 2 mg/mL lypopolysaccaride (LPS) for 24 h. Then, HUVEC were tested for the expression of Vascular Cell Adhesion Molecule(VCAM)-1 by enzyme immunoassays (EIA), and U937 supernatants were tested for the release of matrix metalloproteinases (MMP)-9 by gelatine zymography. Results: All chemically synthesized polyphenol tested reduced, in a concentration dependent manner, the stimulated expression of VCAM-1 in LPS triggered HUVEC. In the same conditions, lipophilic fraction extracted from both Primitivo and Negroamaro wines as well as the corresponding de-alcoholised wines exhibited an even higher inhibition of VCAM-1 expression (up to 50-70 fold higher than those expected on the base of individual polyphenol content). Similarly, either Primitivo and Negroamaro polyphenolic extracts reduced the PMA stimulated release of MMP-9 by PMA triggered U937.Conclusions: In this study different class of polyphenols from Primitivo and Negramaro red wines were characterised. These fractions, significantly inhibited the monocyte-derived extracellular matrix proteases release as well as the endothelial expression of athero-adhesion molecules. These inhibitory effects could explain, at least in part, the anti-inflammatory and cardio-protective properties of red wine polyphenols.

Flavonoids are a large family of plant polyphenolic secondary metabolites. Although they are widespread throughout the plant kingdom, some flavonoid classes are specific for only a few plant species. Due to their presumed health benefits there is growing interest in the development of food crops with tailor-made levels and composition of flavonoids, designed to exert an optimal biological effect. In order to explore the possibilities of flavonoid engineering in tomato fruits, we have targeted this pathway towards novel classes of potentially healthy flavonoids. Using a structural gene (encoding a grape stilbene synthase), we were able to produce transgenic tomatoes accumulating new stilbene-related phytochemicals. Afterwards, we performed a breeding programme taking advantage of the ability of some transcription factors (MYB12, Delila and Rosea) to strongly activate the expression of several phenylpropanoid biosynthetic genes and provide high rates of metabolic flux for different classes of polyphenols. Biochemical analyses showed that these new tomato lines were able to accumulate high levels of specific classes of polyphenols such as stilbenes, flavonols and anthocyanins. We demonstrated that, due to the presence of the novel phytochemicals, the transgenic tomato fruits displayed a significantly higher antioxidant profile. Our data show that a combination of biosynthetic and regulatory genes together with the availability of natural tomato varieties could provide novel insights into genetic and biochemical regulation of the flavonoid pathway in this worldwide important vegetable.

There is a growing interest in producing food plants with increased amounts of antioxidants because of their potential health benefits. In particular polyphenolic secondary metabolites, such as flavonoids and stilbenes, have been investigated for their ignificant antioxidant activity, important both for plant physiology and human nutrition. With the aim of generating plants and fruits with increased antioxidant capability and a wider health benefits, the tomato flavonoid route was modified through the over-expression of the grape stilbene synthase cDNA under a constitutive promoter (Giovinazzo et al., 2005) and a tissue specific, TomLoxB, promoter (D'Introno et al., 2009). The expression of the StS gene in tomato tissues resulted in the synthesis of new compounds, identified as trans-resveratrol and trans-resveratrol-glucopyranoside with a different tissue specificity, both depending on the promoter and the availability of precursors in ripening tissues (Nicoletti et al., 2007). The level synthesis of compounds belonging to different class of polyphenols (i.e. chlorogenic acid, naringenin, rutin, and quercetin), were monitored in both transgenic and wild type fruits tissues at different stages of maturity. Furthermore, we wanted to analyse whether the synthesis of resveratrol in tomato fruit affected the redox status of transformed tomato tissues. The results indicated that the stilbenes and the flavonoids found in transgenic plants were mainly detected as conjugates. This could increase the bioavailability of these compounds. Hence, the structure of the stilbenes synthesized in the transformed tomato plants is of considerable interest from a nutritional point of view. Moreover, the synthesis of resveratrol affected the levels of other antioxidants. In particular, ascorbate and glutathione pool increased in the transformed tissues, proportionally to the amount of the resveratrol. Noteworthy, the transformed fruits also showed increased activities of different enzymes involved in redox homeostasis, (such as ASC - redox enzymes and catalase) which finally resulted in a lower levels of lipid peroxidation. The alteration induced by the presence of resveratrol on cell metabolism will be discussed.

Introduction: Wine grape pomace (WGP), the byproduct of wine making, is a good source of polyphenols, dietary fiber, metals, and may be incorporated into various food products as a functional ingredient. This study investigated the effects of different drying methods and storage up to 6 months on the retention of polyphenols and essential metals in two types of red WGP skin. The biological properties of WGP polyphenol extracts were also analysed in inflamed human cultured endothelial cells.Material & Methods: Antioxidant activity, total phenols content, polyphenol and anthocyanin analyses were carried out (Gerardi et al., 2015; D'Introno et al. 2009). Metal concentrations were measured using the ICP-AES technique (Scortichini et al., 2018). Biological activity of WGP polyphenol extracts were evaluated through the expression of endothelial adhesion molecules (VCAM-1, ICAM-1 and E-Selectin) by EIA (Calabriso et al., 2016).Results: The different drying methods tested showed similar polyphenol contents and antioxidant activity of two WGP skin. The WGP polyphenol extracts significantly reduced the endothelial adhesion molecules expression in endothelial cells stimulated by lipopolysaccharide.Conclusion: Our findings appreciate the grape skins pomace as rich source of bioactive compounds with antioxidant and anti-inflammatory activity and suggest their exploitation as functional ingredients.

Dietary polyphenols are associated with a wide range of health benefits, protecting against chronic diseases and promoting healthy aging. Dietary polyphenols offer a complementary approach to the treatment of inflammatory bowel diseases (IBDs), a group of common chronic intestinal inflammation syndromes for which there is no cure. Tomato is widely consumed but its content of polyphenols is low. We developed a tomato variety, Bronze, enriched in three distinct classes of polyphenols: flavonols, anthocyanins, and stilbenoids. Using Bronze tomatoes as a dietary supplement as well as Indigo (high anthocyanins and flavonols), ResTom (high stilbenoids) and wild-type tomatoes, we examined the effects of the different polyphenols on the host gut microbiota, inflammatory responses, and the symptoms of chronic IBD, in a mouse model. Bronze tomatoes significantly impacted the symptoms of IBD. A similar result was observed using diets supplemented with red grape skin containing flavonols, anthocyanins, and stilbenoids, suggesting that effective protection is provided by different classes of polyphenols acting synergistically.

Curcumin is a natural hydrophobic polyphenol found in the powdered rhizomes of Curcuma longa. Due to its capacity to interfere with many signalling pathways, it has been shown that curcumin has potential beneficial pharmacological effects including antioxidant, anti-inflammatory, anticarcinogenic properties. However, the use of curcumin is fairly restricted because of its poor water solubility, low bioavailability, inadequate tissue absorption and degradation at alkaline pH. In the present contribution, we first verified the anti-proliferative effects of natural curcuminoids towards two different cell lines derived from an ovarian and a breast adenocarcinoma cancer. Later, curcuminoids were successfully encapsulated into reconstituted oil bodies. Once encapsulated into the triacylglycerol cores of the reconstituted oil bodies, curcumin, the most hydrophobic and active of the three curcuminoids, was better stabilized in comparison with albumin stabilization. Oil body encapsulated curcuminoids showed the same effects on cancer cell viability as the free drug, confirming the great potential of natural oil bodies as micro/nano-capsules in drug delivery applications.

Flavonoids are a large family of plant polyphenolic secondary metabolites. Although they are widespread throughout the plant kingdom, some flavonoid classes are specific for only a few plant species. Due to their presumed health benefits there is growing interest in the development of food crops with tailor-made levels and composition of flavonoids, designed to exert an optimal biological effect. In order to explore the possibilities of flavonoid engineering in tomato fruits, we have targeted this pathway towards novel classes of potentially healthy flavonoids. Using a structural flavonoid gene (encoding a grape stilbene synthase), we were able to produce transgenic tomatoes accumulating new stilbene-related phytochemicals. Afterwards, we performed a breeding programme taking advantage of the ability of some transcription factors (MYB12, Delila and Rosea) to strongly activate the expression of several phenylpropanoid biosynthetic genes and provide high rates of metabolic flux for different classes of polyphenols. Biochemical analyses showed that these new tomato lines were able to accumulate high levels of specific classes of polyphenols such as stilbenes, flavonols and anthocyanins. We demonstrated that, due to the presence of the novel phytochemicals, the transgenic tomato fruits displayed a significantly higher antioxidant profile. Our data show that a combination of biosynthetic and regulatory genes together with the availability of natural tomato varieties could provide novel insights into genetic and biochemical regulation of the flavonoid pathway in this worldwide important vegetable.

Grape berries polyphenols are mainly synthesized in the skin tissues and seeds and they are extracted during the winemaking process. These substances have a potentially positive effect, on human health, thus giving to grape and red wine "functional properties" that can contribute to prevent a number of human illness. Nevertheless, the research community is showing that the real effect is a result of a combination of different factors notably daily intake, bioavailability, or in vivo antioxidant activity that are yet to be resolved. Viticulture and winemaking practices, determine the concentration of polyphenols in grape and wine. To date, reduced knowledge is existing on the effects of different yeast strains on the final concentration of polyphenols in red wine. We summarize the recent findings concerning the effects of polyphenols on human chronic disease and the future directions for research to increase the amount of these compounds in wine.

Polyphenols represent a great variety of compounds occurring in fruits, vegetables and plant-derived products. Dietary polyphenols have been found displaying several biological properties, such as anti-inflammatory, antioxidant and anti-aging activities, cardiovascular and neuro-protection, and reduction of the risk of intestinal diseases. The bio-efficacy of polyphenols is tightly linked to their bioavailability, to structural complexity and composition of food matrix in which they are present. Since most of the polyphenols are naturally stored in food matrices as glycosylated and/or variously decorated forms, they need an intestinal bio-conversion in more absorbable forms. Recent findings are highlighting the polyphenols-gut microbiota interplay in the health benefits linked to these compounds. Furthermore, the prebiotic-like activities of polyphenols on microbiota and their potential use in preventive/therapeutic strategies for gastrointestinal disorders are recently emerging.

Halloysite nanotubes (HNTs) are efficient nano-containers capable of entrapping a range of active agents within the inner lumen, followed by their retention and slow release. Halloysite is a green environmentally friendly object available in commercial quantities. The lumen of the halloysite tube accomodates globular protein diameters, allowing their entrapment within the inner lumen of the halloysite while retaining their activity for use in biocatalysis. In this work a combination of high resolution imaging technique such as TEM, SEM and SFM have been employed to elucidate the structure. We have investigated their visco-elastic properties by force-indentation measurements and performed cytotoxicity tests utilizing neoplastic cell lines (breast and cervical cancer cells). Furthermore their uptake has been confirmed by Confocal Microscopy after their functionalisation with fluorescence molecules. The results indicate that halloysite nanotubes have been readly uptaken by neoplastic cells and exhibit a high level of biocompatibility.

Phenolic coumpounds present in grape berries are extracted from the skin, seeds and flesh during the winemaking process. These substances have a potentially positive effect, on human health, thus giving to red wine "bioactive properties" that an contribute to decrease the incidence of atherosclerosis, cancer, neurodegenerative diseases. Grape variety, quality, climate, geographical origin and phythopatologies can affect the quality and quantity of phenolic compounds that accumulate in the cells. Viticulture and vinification practice, which varies in different countries, determine the concentration of phenolic compounds in wine. During the vinemaking process numerous parameters, such as temperature, presence of seeds and berry skins and addition of enzymes, have been reported to affect the extraction of phenolic compounds during grape must fermentation. Conversely, poor knowledge is available on the effects of different yeast strains on the final concentration of polyphenols in red wine. Indeed, yeasts enzymatic activity are likely to influence the extraction of phenolic substances from grape tissues. With the aim to evaluate the existence of a potential correlation between polyphenols content, antioxidant capacity of wine and yeast starter cultures, three selected autochthonous strains of Saccharomyces cerevisiae, were used to inoculate large scale must fermentation. At the end of the process, the experimental wines obtained were evaluated for the content of different class of polyphenols and antioxidant capacity. The statistical analysis of obtained results revealed a significant influence of the autochthonous strain on the concentration of several class of polyphenols and total antioxidant capacity in the produced wines. The different strain influence on the analysed parameters assumes a technological significance and it strongly contribute in determining the final quality of the wine by influencing "functional parameters" important for the preservation of human health.

Colon is exposed to a number of prooxidant conditions and several colon diseases are associated with increased levels of reactive species. Polyphenols are the most abundant antioxidants in the diet, but to date no information is available about their absorption and potential intracellular antioxidant activity on colon epithelial cells. The work was addressed to study the intracellular antioxidant activity of red grape polyphenolic extracts on rat colon epithelium experimentally exposed to prooxidant conditions. The experimental model chosen was represented by freshly isolated colon explants, which closely resemble the functional, and morphological characteristics of the epithelium in vivo. The study was carried out by in situ confocal microscopy observation on CM-H2DCFDA charged explants exposed to H2O2 (5, 10, and 15 min). The qualitative and quantitative polyphenolic composition of the extracts as well as their in vitro oxygen radical absorbing capacity (ORAC) was determined. The incubation of the explants with the polyphenolic extracts for 1 h produced a significant decrease of the H2O2 induced fluorescence. This effect was more pronounced following 15 min H2O2 exposure with respect to 5 min and it was also more evident for extracts obtained from mature grapes, which showed an increased ORAC value and qualitative peculiarities in the polyphenolic composition. The results demonstrated the ability of red grape polyphenols to cross the plasma membrane and exert a direct intracellular antioxidant activity in surface colonocytes, inducing a protection against pro-oxidant conditions. The changes in the polyphenol composition due to ripening process was reflected in a more effective antioxidant protection.

The plant phenol trans-resveratrol, which is mainly found in grape, displays a wide range of biological effects. A cell suspension culture was developed from calli of grape leaves of Vitis vinifera cv. Negramaro in order to study the bioproduction of resveratrol. The effects of a number of secondary plant metabolism elicitors, namely chitosan, methyl jasmonate, jasmonic acid, coronatine, and 12-oxo-phytodienoic acid, were tested on this cell suspension culture. The identification and quantification of stilbenes was achieved with high performance liquid chromatography, with both spectrophotometric and mass spectrometric detection. Of the tested elicitors, methyl jasmonate was the most effective in inducing the biosynthesis of approximately 4 mg g-1 dry weight (about 60 mg L-1) of resveratrol. Conversely, 12-oxo-phytodienoic acid, jasmonic acid, and coronatine were able to trigger the synthesis of approximately 20 mg g-1 dry weight (200-210 mg L-1) of viniferins. Taken together, our results show for the first time different modulatory effects of closely-related jasmonates on stilbene biosynthesis.

Resveratrol, a plant phenolic compound, is accumulated in grape and red wine, but it is not widely distributed in other food crops. Our previous works showed that resveratrol biosynthesis can be induced in tomato plant (Lycopersicum esculentum Mill.) via the heterologous expression of a grape (Vitis vinifera L.) stilbene synthase cDNA under the control of a constitutive (CaMV 35S) or a fruit specific (TomLoxB) promoter. Resveratrol synthesising fruits showed an increase in the levels of ascorbate and glutathione, the soluble antioxidants of primary metabolism and the total antioxidant activity. These results paralleled with a higher capability of transgenic fruit to counteract the pro-inflammatory effects of phorbol ester in mocyte-macrophages, via the inhibition of induced cyclooxygenase-2, and indicated the potential of our approach to improve the nutritional value of tomato. Since the levels of resveratrol detected in transgenic fruit were limited to a maximum of 50 ug/g fresh weight, we started a breeding programme to increase the content of this metabolite.We took advantage of the ability of an Arabidopsis myb-type transcription factor (AtMYB12) to strongly activate the expression of several phenylpropanoid biosynthetic genes that act upstream of stilbene synthase, suggesting that AtMYB12 can provide high rates of metabolic flux for resveratrol biosynthesis. To further increase the level of resveratrol, we have introduced in the breeding programme a spontaneous flavanone 3-hydroxylase tomato mutant. The presence of this mutation reduces the amount flavonols and should therefore enhance substrate availability to stilbene synthase. Preliminary data show that a combination of biosynthetic and regulatory genes together with the availability of natural tomato varieties is a powerful strategy for plant metabolic engineering.

Purpose: The aim of the study was to evaluate the vascular anti-inflammatory effects of polyphenolic extracts from two typical South Italy red wines, the specific contribution of individual polyphenols and the underlying mechanisms of action.Methods: Human endothelial cells were incubated with increasing concentrations (1-50 ?g/mL) of Primitivo and Negroamaro polyphenolic extracts (PWPE and NWPE, respectively) or pure polyphenols (1-25 ?mol/L), including hydroxycinnamic acids (p-coumaric, caffeic and caftaric acids), flavonols (kaempferol, quercetin, myricetin) or stilbenes (trans-resveratrol, trans-piceid) before stimulation withlipopolysaccharide. Through multiple assays, we analyzed the endothelial-monocyte adhesion, theendothelial expression of adhesion molecules (ICAM-1, VCAM-1 and E-Selectin), monocytechemoattractant protein-1 (MCP-1) and macrophage colony-stimulating factor (M-CSF), as well as ROSintracellular levels and the activation of NF-?B and AP-1.Results: Both PWPE and NWPE, already at 1 ?g/mL, inhibited monocyte adhesion to stimulated endothelial cells, a key event in triggering vascular inflammation. They down-regulated the expression of adhesion molecules, ICAM-1, VCAM-1, E-Selectin, as well as MCP-1 and M-CSF, at mRNA and protein levels. All polyphenols reduced intracellular ROS, and everything, except caftaric acid, inhibited the endothelial expression of adhesion molecules and MCP-1, although with different potency. Flavonols and resveratrol significantly reduced also the endothelial expression and release of M-CSF. The decrease in endothelial inflammatory gene expression was related to the inhibition of NF-?B and AP-1 activation but not to intracellular oxidative stress.Conclusions: This study showed multiple anti-inflammatory and anti-atherosclerotic properties of red wine polyphenolic extracts and indentified specific bioactive polyphenols which could counteract inflammatory diseases including atherosclerosis.

A novel strategy to induce parthenocarpy in tomato fruits by the induction of resveratrol biosynthesis inflower tissues was exploited. Two transgenic tomato lines were considered: a higher resveratrolproducing(35SS) line, constitutively expressing a grape stilbene synthase cDNA, and a lower resveratrolproducing(LoxS) line, expressing stilbene synthase under a fruit-specific promoter. The expression of thestilbene synthase gene affected flavonoid metabolism in a different manner in the transgenic lines, and inone of these, the 35SS line, resulted in complete male sterility. Resveratrol was synthesised either in 35SSor LoxS tomato flowers, at an even higher extent (about 8e10 times) in the former line. We furtherinvestigated whether stilbene synthase expression may have resulted in impaired naringenin accumulationduring flower development. In the 35SS flowers, naringenin was significantly impaired by about50%, probably due to metabolic competition. Conversely, the amount of glycosylated flavonols increasedin transgenic flowers, thereby excluding the diminished production of flavonols as a reason forparthenocarpy in tomato. We further investigated whether resveratrol synthesis may have resultedchanges to pollen structure. Microscopic observations revealed the presence of few and abnormal flakelikepollen grains in 35SS flowers with no germination capability. Finally, the analysis of coumaric andferulic acids, the precursors of lignin and sporopollenin biosynthesis, revealed significant depletion ofthese compounds, therefore suggesting an impairment in structural compounds as a reason for pollenablation. These overall outcomes, to the best of our knowledge, reveal for the first time the major roledisplayed by resveratrol synthesis on parthenocarpy in tomato fruits.

Oil bodies (OBs) are specialised organelles ubiquitously detected in plant oil seeds, which serve as lipid storage compartments. OBs consist of a hydrophobic core of triacylglycerol (TAGs), surrounded by a monolayer of phospholipids (PLs) embedded with some specific proteins with a size ranging from 0.5 to 2 mu m. In this work, we report an easy method to reconstitute OBs starting from their constituents and to encapsulate lipophilic molecules, i.e. the fluorescent fluorescein isothiocyanate (FITC) and carboxyfluorescein (CF), into reconstituted OBs. This methods allowed us to produce OBs 4- to 10-fold smaller (50-200 nm) than the native one and to obtain a good recovery (about 40%) of both the fluorescent compounds used in the present work. The properties of reconstituted OBs were investigated by a combination of Brewster angle microscopy, scanning force microscopy, zeta-potential techniques. OBs were stable and formed ordered monolayers when patterned on hydrophobic substrates whereas they showed a higher tendency to aggregate into larger, coalescing OBs when were deposited onto hydrophilic substrates or at the air/water interface. Furthermore, we verified the uptake of FITC-loaded OBs by the MCF-7 breast cancer cell line. Our results indicated that OBs could be envisaged as novel carriers to deliver hydrophobic bioactive compounds

Red wine is a treasured source of polyphenols, such as stilbenes, flavonols and hydroxycinnamic acids, depending on different factors, such as grape cultivar and wine-making practices. The red wine polyphenols (RWPs) have long been associated to reduced mortality and morbidity from cardiovascular diseases and cancer. Since matrix metalloproteinase (MMP) activity is involved in extracellular matrix degradation, a crucial step both in inflammation and cell migration, aim of this study was to examine the effects of specific classes of polyphenols from Primitivo and Negroamaro grapes and wines produced in the Apulian region (Southern Italy), on the release and activity of MMP-9 in human monocytoid cells.U937 monocitoid cells were pre-treated with increasing concentrations of polyphenolic extracts (PE) of Primitivo and Negramaro grape and wine, before stimulation with 30 nM phorbol myristate acetate (PMA) for 24 h. The release and activity of MMP-9 in culture medium was tested by ELISA and zymography, respectively. Both Primitivo and Negroamaro PE exhibited a concentration-dependent inhibition of MMP-9 release and activity, without any reduction in cell viability. The inhibitory effect was higher than that produced by individual chemically synthesized polyphenols.. In this study, the PE from Primitivo and Negramaro grapes and red wines were characterized, and shown to significantly and synergistically inhibit the monocyte-derived release and activity of extracellular matrix proteases. These inhibitory effects could explain, at least in part, the anti-inflammatory and anti-cancerogenic properties of RWP.

Red wine polyphenols have long been associated to a reduced morbidity and mortality from atherosclerotic disease in part by reducing inflammation, however the effects of different polyphenolic compounds are not clear. The aim of this study was to evaluate the anti-inflammatory and anti-oxidant effects of polyphenolic extracts from two red wines typical of Southern Italy, and to dissect the specific contribute of hydroxycinnamic acids, flavonols and stilbenes and to investigate underlying mechanisms of action in endothelial cells.Methodology and Results: Through multiple assays, we showed that both Primitivo and Negroamaro Wine Polyphenolic Extracts significantly inhibited monocyte adhesion to endothelium, endothelial expression of adhesion molecules, monocyte chemoattractant protein-1 and macrophage colony-stimulating factor as well as intracellular oxidative stress and the activation of NF-?B and AP-1, without any reduction in cell viability. We also showed that all tested polyphenols exhibited a strong antioxidant action in stimulated human endothelial cells but not all significantly exhibited anti-inflammatory activity which in turn was related to the inhibition of NF-?B and AP-1 activation.Conclusions: The present study provides new evidences on specifc anti-inflammatory and anti-atherosclerotic role of red wine polyphenols including hydroxycinnamic acids, flavonols and stilbenes.

Matrix metalloproteinases (MMPs) are endopeptidases responsible for the hydrolysis of various components of extracellular matrix. MMPs, namely gelatinases MMP-2 and MMP-9, contribute to the progression of chronic and degenerative diseases. Since gelatinases' activity and expression are regulated by oxidative stress, we sought to evaluate whether supplementation with polyphenol-rich red grape skin extracts modulated the matrix-degrading capacity in cell models of vascular inflammation. Human endothelial and monocytic cells were incubated with increasing concentrations (0.5-25 ?g/mL) of Negroamaro and Primitivo red grape skin polyphenolic extracts (NSPE and PSPE, respectively) or their specific components (0.5-25 ?mol/L), before stimulation with inflammatory challenge. NSPE and PSPE inhibited, in a concentration-dependent manner, endothelial invasion as well as the MMP-9 and MMP-2 release in stimulated endothelial cells, and MMP-9 production in inflamed monocytes, without affecting tissue inhibitor of metalloproteinases (TIMP)-1 and TIMP-2. The matrix degrading inhibitory capacity was the same for both NSPE and PSPE, despite their different polyphenolic profiles. Among the main polyphenols of grape skin extracts, trans-resveratrol, trans-piceid, kaempferol and quercetin exhibited the most significant inhibitory effects on matrix-degrading enzyme activities. Our findings appreciate the grape skins as rich source of polyphenols able to prevent the dysregulation of vascular remodelling affecting degenerative and inflammatory diseases.

There is a growing interest in producing food plants with increased amounts of antioxidants because of their potential health benefits. Polyphenolic secondary metabolites, such as flavonoids and stilbenes, have been investigated for their significant antioxidant activity, important both for plant physiology and human nutrition. With the aim to verify whether the synthesis of resveratrol in tomato fruit affected the redox status of transformed tomato tissues, in the present work we took advantage of the availability of two different transgenic tomato lines in which a grape stilbene synthase gene was expressed under a constitutive or a tissue specific promoter, respectively. The induction of resveratrol synthesis in tomato affected the redox status of transformed tomato fruit. In particular, ascorbate and glutathione pool increased, significantly and proportionally to the amount of the resveratrol synthesised in transformed tissues. Noteworthy, the increase in ascorbate and glutathione pool paralleled with a significant increase in the activities of the main enzymes involved in redox homeostasis, i.e. catalase, ascorbate peroxidase as well the enzymes responsive for ascorbate recycling. Furthermore, lipoxygenase activity and levels of polyunsaturated fatty acids hydroperoxides, were reduced in fruit tissues from transgenic tomato lines. The synthesis of resveratrol, together with the global redox status had an impact on the total antioxidant activity of transgenic fruits. Again the total antioxidant capability increased proportionally to the amount of synthesised resveratrol with the most remarkable increase recorded from the lipophilic-resveratrol containing-fraction of transgenic fruits. Overall, our results pointed a higher nutritional value of resveratrol synthesizing tomato fruits.

The plant polyphenol trans-resveratrol (3, 5, 4?-trihydroxystilbene) mainly found in grape, peanut and otherfew plants, displays a wide range of biological effects.Numerous in vitro studies have described various biologicaleffects of resveratrol. In order to provide more informationregarding absorption, metabolism, and bioavailability ofresveratrol, various research approaches have been performed,including in vitro, ex vivo, and in vivo models. Inrecent years, the induction of resveratrol synthesis in plantswhich normally do not accumulate such polyphenol, hasbeen successfully achieved by molecular engineering.Inthis context, the ectopic production of resveratrol has beenreported to have positive effects both on plant resistance tobiotic stress and the enhancement of the nutritional value ofseveral widely consumed fruits and vegetables. The metabolicengineering of plants offers the opportunity to changethe content of specific phytonutrients in plant - derivedfoods. This review focuses on the latest findings regardingon resveratrol bioproduction and its effects on the preventionof the major pathological conditions in man.

Carcinoma progression is associated with the loss of epithelial features, and the acquisitionof a mesenchymal phenotype by tumour cells. Herein we show that exposure of MCF-7cells to epidermal growth factor (EGF) resulted in morphological alterations characteristicof epithelial-to-mesenchymal transition (EMT). EGF treatment resulted in increased motilityalong with an up-regulation of transcription factors Slug, Zeb1, Zeb2, and mesenchymalmarkers Vimentin and N-cadherin.Treatment of MCF-7 cells with a combined stimulation of EGF and resveratrol, a naturallyoccurring stilbene with antitumor properties, failed to alter cell morphology, motility andoverexpression of EMT markers induced by EGF. Using specific chemical inhibitors, wedemonstrated that EGF-induced EMT is mediated by extracellular signal-regulated kinase1/2 (ERK 1/2) signalling pathway and that resveratrol is able to repress EGF-induced ERKactivation.In summary, these data provide new evidence of the inhibitory effect of resveratrol onEGF-induced EMT cell transformation.