Variety and year: Two key factors on amino acids and biogenic amines content in grapes.

Amino acids have proved to play a key role in the development of volatile compounds present in wine with determining repercussions on the final wine bouquets. Biogenic amines originate from the chemical transformations of amino acids found in various foods, a phenomenon that has given rise to several health-related concerns among consumers. In the present research, the evaluation of two of the most influential factors: variety (genetic) and year (climatic conditions) on these compounds in grapes has been performed. Eight Vitis vinifera varieties have been collected during three years and the content of nineteen amino acids, two biogenic amines, and the ammonium ion has been quantified using the HPLC-PDA technique. The genetic factor has proved to be an influential variable (p-value < 0.05) with mean values of amino acids ranging from 896.89 to 1713.79 mg/L and of biogenic amines ranging from 10.61 to 22.28 mg/L. The climatic conditions have shown to be an influential factor as well (p-value < 0.05), being the low temperatures and rainfall and the high solar radiation favour the development of the amino acid and avoid biogenic amines accumulation in grapes.

Holistic understanding of the response of grapevines to foliar application of seaweed extracts.

Viticulture is highly dependent on phytochemicals to maintain good vineyard health. However, to reduce their accumulation in the environment, green regulations are driving the development of eco-friendly strategies. In this respect, seaweeds have proven to be one of the marine resources with the highest potential as plant protective agents, representing an environmentally-friendly alternative approach for sustainable wine production. The current work follows an interdisciplinary framework to evaluate the capacity of Ulva ohnoi and Rugulopteryx okamurae seaweeds to induce defense mechanisms in grapevine plants. To our knowledge, this is the first study to evaluate Rugulopteryx okamurae as a biostimulator . This macroalgae is relevant since it is an invasive species on the Atlantic and Mediterranean coast causing incalculable economic and environmental burdens. Four extracts (UL1, UL2, RU1 and RU2 developed from Ulva and Rugulopteryx, respectively) were foliar applied to Tempranillo plants cultivated under greenhouse conditions. UL1 and RU2 stood out for their capacity to induce defense genes, such as a PR10, PAL, STS48 and GST1, mainly 24 hours after the first application. The increased expression level of these genes agreed with i) an increase in trans-piceid and trans-resveratrol content, mainly in the RU2 treated leaves, and, ii) an increase in jasmonic acid and decrease in salicylic acid. Moreover, an induction of the activity of the antioxidant enzymes was observed at the end of the experiment, with an increase in superoxide dismutase and catalase in the RU2-treated leaves in particular. Interestingly, while foliar fungal diversity was not influenced by the treatments, alga extract amendment modified fungal composition, RU2 application enriching the content of various groups known for their biocontrol activity. Overall, the results evidenced the capacity of Rugulopteryx okamurae for grapevine biostimulation, inducing the activation of several secondary metabolite pathways and promoting the abundance of beneficial microbiota involved in grapevine protection. While further studies are needed to unravel the bioactive compound(s) involved, including conducting field experiments etc., the current findings are the first steps towards the inclusion of Rugulopteryx okamurae in a circular scheme that would reduce its accumulation on the coast and benefit the viticulture sector at the same time.

Sulfur dioxide-free Verdejo wines through the use of a pure stilbene extract: exploring possible synergistic effect with glutathione.

BACKGROUND: The search for alternatives to sulfur dioxide (SO2 ) in white wine production is a tough challenge for the oenological industry. The current article investigates the effectiveness of several preservatives in protecting Verdejo white wines. The following wines were elaborated: without preservative (CT wines), with SO2 (SO2 wines), with stilbene-enriched extract (ST99 wines), with glutathione (GSH wines) and finally with the combination stilbene extract plus glutathione (ST99-GSH wines). RESULTS: ST99 extract preserved the wines from malolactic fermentation (MLF), maintained the quality parameters of the wines, but affected their color from the beginning of the process. Synergistic effects between ST99 and GSH were found for oxygen consumption rates, related to oxidation processes. The content of amino acids and biogenic amines was slightly affected by the treatments, but the quality or safety of the wines was not compromised. However, after 12 months of ageing in bottle, the SO2 wines showed the highest score in the sensory analysis. The reasons for the evolution of treated wines and the implications of this study for maintaining the quality of free SO2 white wines are discussed. CONCLUSION: ST99 may be proposed as an alternative to SO2 in wines to be consumed in the short term. Its combination with GSH does not prolong its shelf-life. © 2022 Society of Chemical Industry.

Wood Waste from Fruit Trees: Biomolecules and Their Applications in Agri-Food Industry.

In the European Union (EU), a total of 11,301,345 hectares are dedicated to the cultivation of fruit trees, mainly olive orchards, grapevines, nut trees (almond, walnut, chestnut, hazelnut, and pistachio), apple and pear trees, stone fruit trees (peach, nectarine, apricot, cherry, and plum), and citrus fruit trees (orange, clementine, satsuma, mandarin, lemon, grapefruit, and pomelo). Pruning these trees, together with plantation removal to a lesser extent, produces a huge amount of wood waste. A theoretical calculation of the wood waste in the European Union estimates approximately 2 and 25 million tons from wood plantation removal and pruning, respectively, per year. This wood waste is usually destroyed by in-field burning or crushing into the soil, which result in no direct economic benefits. However, wood from tree pruning, which is enriched in high added-value molecules, offers a wide spectrum of possibilities for its valorization. This review focuses on the contribution of wood waste to both sustainability and the circular economy, considering its use not only as biomass but also as a potential source of bioactive compounds. The main bioactive compounds found in wood are polyphenols, terpenes, polysaccharides, organic compounds, fatty acids, and alkaloids. Polyphenols are the most ubiquitous compounds in wood. Large amounts of hydroxytyrosol (up to 25 g/kg dw), resveratrol (up to 66 g/kg dw), protocatechuic acid (up to 16.4 g/kg), and proanthocyanins (8.5 g/kg dw) have been found in the wood from olive trees, grapevines, almond trees and plum trees, respectively. The bioactivity of these compounds has been demonstrated at lower concentrations, mainly in vitro studies. Bioactive compounds present antioxidant, antimicrobial, antifungal, biostimulant, anti-inflammatory, cardioprotective, and anticarcinogenic properties, among others. Therefore, wood extracts might have several applications in agriculture, medicine, and the food, pharmaceutical, nutraceutical, and cosmetics industries. For example, olive tree wood extract reduced thrombin-induced platelet aggregation in vitro; grapevine tree wood extract acts a preservative in wine, replacing SO2; chestnut tree wood extract has antifungal properties on postharvest pathogens in vitro; and stone tree wood extracts are used for aging both wines and brandies. Moreover, the use of wood waste contributes to the move towards both a more sustainable development and a circular economy.

Wine Polyphenol Content and Its Influence on Wine Quality and Properties: A Review.

Wine is one of the most consumed beverages around the world. It is composed of alcohols, sugars, acids, minerals, proteins and other compounds, such as organic acids and volatile and phenolic compounds (also called polyphenols). Polyphenols have been shown to be highly related to both (i) wine quality (color, flavor, and taste) and (ii) health-promoting properties (antioxidant and cardioprotective among others). Polyphenols can be grouped into two big families: (i) Flavonoids, including anthocyanidins, flavonols, flavanols, hydrolysable and condensed tannins, flavanones, flavones and chalcones; and (ii) Non-flavonoids, including hydroxycinnamic acids, hydroxybenzoic acids, stilbenes, tyrosol and hydroxytyrosol. Each group affects in some way the different properties of wine to a greater or a lesser extent. For that reason, the phenolic composition can be managed to obtain singular wines with specific, desirable characteristics. The current review presents a summary of the ways in which the phenolic composition of wine can be modulated, including (a) invariable factors such as variety, field management or climatic conditions; (b) pre-fermentative strategies such as maceration, thermovinification and pulsed electric field; (c) fermentative strategies such as the use of different yeasts and bacteria; and (d) post-fermentative strategies such as maceration, fining agents and aging. Finally, the different extraction methods and analytical techniques used for polyphenol detection and quantification have been also reviewed.

Impact of Sequential Inoculation with the Non-Saccharomyces T. delbrueckii and M. pulcherrima Combined with Saccharomyces cerevisiae Strains on Chemicals and Sensory Profile of Rosé Wines.

Controlled inoculations of non-Saccharomyces yeasts are becoming increasingly used to produce high-quality wines due to their enological potential. In this study, we evaluated the impact of sequential inoculation with the commercial non-Saccharomyces yeasts (Torulaspora delbrueckii and Metschnikowia pulcherrima) in combination with Saccharomyces cerevisiae on the chemical and sensory profile of rosé wines. Sequential inoculation with T. delbrueckii produced wines with an overall reduction in esters, mainly explained by the lower concentrations of ethyl esters of medium-chain fatty acids and isoamyl acetate. The lower ester concentrations of these wines were related to a reduction in fruity descriptors. An increase was observed, however, in other minor esters such as cinnamates and ethyl esters of branched acids. Zinc, ethyl isobutyrate, and ethyl dihydrocinnamate were selected as potential markers for this fermentation strategy. Sequential inoculation with M. pulcherrima resulted in rosé wines with an enhanced ester profile, reduced acetaldehyde, and increased anthocyans and tannins. Compared to the control wines fermented with S. cerevisiae, the changes observed in these wines were far subtler, especially for the volatile profile, sensory characteristics, and color parameters, with isobutyl hexanoate and isoamyl butyrate being selected as potential markers.

Cytotoxicity studies of a stilbene extract and its main components intended to be used as preservative in the wine industry.

The use of stilbenes has been proposed as an alternative to sulfur dioxide in wine. Provided the feasibility from a technological approach, the cytotoxicity of an extract from grapevine shoots containing a stilbene richness of 99% (ST-99 extract) was assessed in the human cell lines HepG2 and Caco-2. In addition, the effects of the main stilbenes found in ST-99, trans-resveratrol and trans-ε-viniferin were studied, as well as its mixture. Similar cytotoxic effects were obtained in the exposures to trans-ε-viniferin, ST-99 and the mixture; however, trans-resveratrol alone exerted less toxicity. When HepG2 cells were exposed to trans-ε-viniferin, ST-99 and the mixture, the mean effective concentration (EC50) were 28.28 ± 2.15, 31.91 ± 1.55 and 29.47 ± 3.54 µg/mL, respectively. However, in the exposure to trans-resveratrol, the EC50 was higher 50 µg/mL. The morphological study evidenced damage at ultrastructural level in HepG2 cells, highlighting the inhibition of cell proliferation and the induction of apoptosis. The type of interaction produced by trans-ε-viniferin and trans-resveratrol mixtures was assessed by an isobologram analysis using the CalcuSyn software, evidencing an antagonist effect. These data comprise a starting point in the toxicological assessment; further studies are needed in this field to assure the safety of the extract ST-99.

Grapevine Cane Extracts: Raw Plant Material, Extraction Methods, Quantification, and Applications.

Grapevine canes are viticulture waste that is usually discarded without any further use. However, recent studies have shown that they contain significant concentrations of health-promoting compounds, such as stilbenes, secondary metabolites of plants produced as a response to biotic and abiotic stress from fungal disease or dryness. Stilbenes have been associated with antioxidant, anti-inflammatory, and anti-microbial properties and they have been tested as potential treatments of cardiovascular and neurological diseases, and even cancer, with promising results. Stilbenes have been described in the different genus of the Vitaceae family, the Vitis genera being one of the most widely studied due to its important applications and economic impact around the world. This review presents an in-depth study of the composition and concentration of stilbenes in grapevine canes. The results show that the concentration of stilbenes in grapevine canes is highly influenced by the Vitis genus and cultivar aspects (growing conditions, ultraviolet radiation, fungal attack, etc.). Different methods for extracting stilbenes from grapevine canes have been reviewed, and the extraction conditions have also been studied, underlining the advantages and disadvantages of each technique. After the stilbenes were extracted, they were analyzed to determine the stilbene composition and concentration. Analytical techniques have been employed with this aim, in most cases using liquid chromatography, coupled with others such as mass spectrometry and/or nuclear magnetic resonance to achieve the individual quantification. Finally, stilbene extracts may be applied in multiple fields based on their properties. The five most relevant are preservative, antifungal, insecticide, and biostimulant applications. The current state-of-the-art of the above applications and their prospects are discussed.

Red Wine Extract Disrupts Th17 Lymphocyte Differentiation in a Colorectal Cancer Context.

SCOPE: Scope: It is well established that immune response and inflammation promote tumoral progression. Immune cells communicate through direct contact or through cytokine secretion, and it is the pro-inflammatory status that will tip the balance toward tumor progression or anti-tumor immunity. It is demonstrated here that a red wine extract (RWE) can decrease inflammation through its action on the inflammasome complex. This study determines whether an RWE could impact other key actors of inflammation, including T helper 17 (Th17) immune cells in particular. METHODS AND RESULTS: Methods and results: Using an RWE containing 4.16 g of polyphenols/liter of wine, it is shown that RWE decreases colorectal cancer cells in vitro and induces a reduction in colorectal tumor growth associated with a decrease in tumor-infiltrating lymphocytes in vivo. The process of T-lymphocyte differentiation in Th17 cells is altered by RWE, as revealed by the decrease in the expression of key actors controlling this process, such as signal transducer and activator of transcription 3 and retinoid acid-related orphan receptor γt. This disruption is associated with an inhibition of inflammatory interleukin 17 secretion. CONCLUSION: The data highlights the major involvement of Th17 immune cells in the biological effects of an RWE.

Multivariate optimization of headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry for the analysis of terpenoids in sparkling wines.

A headspace solid-phase microextraction coupled to gas-chromatography and mass spectrometry (HS-SPME-GC-MS) was developed and validated for the determination of 26 terpenes in sparkling wines. The use of a Box-Behnken experimental design, together with the desirability function D, allowed the extraction conditions of the method to be optimized. The optimal extraction conditions were found at a dilution ratio of 2:3, the addition of 3.5 g of NaCl, an extraction temperature of 46 °C and an extraction time of 52 min, using the DVB/CAR/PDMS fibre. Afterwards, the analytical method was successfully validated in terms of linearity, matrix effect, limit of detection and quantification, precision and accuracy. To test the developed method, 35 commercial sparkling wines from different grape varieties, geographical regions and ageing times were analysed and their terpenoid profile was monitored. The use of multivariate statistical tools made it possible to highlight differences in the samples related to the terpene profile. Finally, the most important compounds involved in the discrimination of the samples were identified by means of iterative variable selection procedures.

In Vitro Toxicity Assessment of Stilbene Extract for Its Potential Use as Antioxidant in the Wine Industry.

The reduction of sulfur dioxide in wine is a consumer's demand, considering the allergic effects that may occur in people who are sensitive to it. Stilbenes are candidates of great interest for this purpose because of their antioxidant/antimicrobial activities and health properties, and also because they are naturally found in the grapevine. In the present study, the in vitro toxicity of an extract from grapevine shoots (with a stilbene richness of 45.4%) was assessed in two human cell lines. Significant damage was observed from 30 µg/mL after 24 h, and 40 µg/mL after 48 h of exposure. Similarly, the ultrastructural study revealed a significant impairment of cell growing. The extract was able to protect cells against an induced oxidative stress at all concentrations studied. In view of the promising results, a more exhaustive toxicological assessment of the extract is needed to confirm the safety of its further use as additive in wine.

Inhibition of VEGFR-2 Phosphorylation and Effects on Downstream Signaling Pathways in Cultivated Human Endothelial Cells by Stilbenes from Vitis Spp.

Stilbenes are phenolic compounds present in different higher plant families that have shown different biological activities, such as antioxidant properties and antitumoral and anti-atherosclerotic effects, among others. Angiogenesis is a key process involved in both cancer and cardiovascular diseases, the vascular endothelial growth factor (VEGF) and its receptor VEGFR-2 being the main triggers. Certain polyphenol compounds, such as flavonoids, have shown a potent capacity to inhibit VEGF and, consequently, angiogenesis. The present work, therefore, aims to evaluate the potential effect of stilbenes on inhibiting VEGF and their subsequent effect on the downstream signaling pathway (PLCγ1, Akt, and eNOS). VEGFR-2 activation was studied through an ELISA assay in the HUVEC line, while the phosphorylation of intracellular downstream proteins PLCγ1, Akt, and eNOS was tested by Western blot. Student's t test was used to determine significant differences between samples. On the one hand, astringin, pallidol, and ω-viniferin showed the lowest IC50 values (2.90 ± 0.27, 4.42 ± 0.67, and 6.10 ± 1.29 µM, respectively) against VEGFR-2 activation. Additionally, VEGF-induced PLCγ1 phosphorylation was significantly inhibited by ε-viniferin, astringin, and ω-viniferin. However, ε-viniferin and pallidol simultaneously enhanced eNOS activation, proving to be via Akt activation in the case of ε-viniferin. For the first time, these data suggest that stilbenes such as astringin, pallidol, ω-viniferin, and ε-viniferin have a potential anti-angiogenic effect and they could be further considered as anti-VEGF ingredients in food and beverages. In addition, ε-viniferin and pallidol significantly allowed eNOS activation and could likely prevent the side effects caused by anti-VEGF hypertension drugs.

Study of Potential Anti-Inflammatory Effects of Red Wine Extract and Resveratrol through a Modulation of Interleukin-1-Beta in Macrophages.

Inflammation has been described as an initiator event of major diseases with significant impacts in terms of public health including in cardiovascular disease, autoimmune disorders, eye diseases, age-related diseases, and the occurrence of cancers. A preventive action to reduce the key processes leading to inflammation could be an advantageous approach to reducing these associated pathologies. Many studies have reported the value of polyphenols such as resveratrol in counteracting pro-inflammatory cytokines. We have previously shown the potential of red wine extract (RWE) and the value of its qualitative and quantitative polyphenolic composition to prevent the carcinogenesis process. In this study, we addressed a new effect of RWE in inflammation through a modulation of IL-1ß secretion and the NLRP3 inflammasome pathway. NLRP3 inflammasome requires two signals, priming to increase the synthesis of NLRP3 and pro-IL-1ß proteins and activation, which activates NLRP3. Inflammasome formation is triggered by a range of substances such as lipopolysaccharide (LPS). Using two different macrophages, one of which does not express the adaptor protein ASC (apoptosis-associated speck-like protein containing a CARD), which is essential to form active inflammasome complexes that produce IL-1ß, we show that RWE decreases IL-1 ß secretion and gene expression whatever line is used. Moreover, this strong reduction of pro-inflammatory IL-1ß is associated with a decrease of NLRP3 and, in J774A, ASC protein expression, which depends on the choice of activator ATP or nigericin.

Effect of a grapevine-shoot waste extract on red wine aromatic properties.

BACKGROUND: The use of a grapevine-shoot extract (VIN) is being studied as an alternative to sulfur dioxide (SO2 ). VIN stabilizes anthocyanins and preserves polyphenolic compounds, and thus improves chromatic wine properties. In this study, selected aroma compounds (esters, C13 -norisoprenoids, oxidation and vine-shoot-related compounds), sensory analysis and the olfactometric profile were determined in the wines treated with VIN at two concentrations. RESULTS: Treatment with VIN hardly modified the content of esters and oxidation-related compounds in the wines. However, the high ß-damascenone and isoeugenol contents and the increase in astringency at tasting in VIN wines were noteworthy, as were some odorant zones. All these were established as VIN markers after the chemometric data analysis. CONCLUSION: These data revealed that only the lowest dose tested may be recommended as a suitable alternative to SO2 . Although some aromatic properties of these wines may change, these changes are not considered to affect the quality of the wines negatively. These results are useful for wineries, which face having to discover the aroma-related processes in the challenge of producing SO2 -free wines without detriment to their sensory properties. © 2018 Society of Chemical Industry.

Sulfur free red wines through the use of grapevine shoots: Impact on the wine quality.

Following a preliminary study to determine the possibility of using a grapevine shoot extract (VIN) as a sustainable alternative to sulfur dioxide (SO2), in this study, the chromatic features, phenolic composition, and sensory analysis of wines treated with VIN at two concentrations were studied during storage in bottle for the first time. The highest differences were found in phenolic compounds after 12months of storage in bottle. The VIN wines had a low content of free anthocyanins and were high in vinyl-pyranoanthocyanins, and B-type vitisins. Consequently, they showed better chromatic characteristics. Moreover VIN, especially at high dose, preserved non-anthocyanin phenolic compounds better than SO2. However, at this high dose some organoleptic properties were affected. VIN, when used at a low dose, is able to preserve wine composition without loss of quality.

Stilbenes from Vitis vinifera L. Waste: A Sustainable Tool for Controlling Plasmopara Viticola.

Stilbene-enriched extracts from Vitis vinifera waste (cane, wood, and root) were characterized by UHPLC-MS. Eleven stilbenes were identified and quantified as follows: ampelopsin A, (E)-piceatannol, pallidol, (E)-resveratrol, hopeaphenol, isohopeaphenol, (E)-ε-viniferin, (E)-miyabenol C, (E)-ω-viniferin, r2-viniferin, and r-viniferin. The fungicide concentration inhibiting 50% of growth of Plasmopara viticola sporulation (IC50) was determined for the extracts and also for the main compounds isolated. r-Viniferin followed by hopeaphenol and r2-viniferin showed low IC50 and thus high efficacy against Plasmopara viticola. Regarding stilbene extracts, wood extract followed by root extract showed the highest antifungal activities. These data suggest that stilbene complex mixtures from Vitis vinifera waste could be used as a cheap source of bioactive stilbenes for the development of natural fungicides.

Daily Preharvest UV-C Light Maintains the High Stilbenoid Concentration in Grapes.

The fact that it is possible to induce stilbenoid synthesis in grapevine (Vitis vinifera) by UV-C light allows the possibility of stimulating grapevine phytoalexin production to increase disease resistance and immunity, and subsequently to limit the use of pesticides in vineyards. UV-C light was applied daily during three days before the harvesting of table grape variety Crimson seedless to study the accumulation of stilbenoid compounds during ripeness. The E-resveratrol concentration was monitored during daily preharvest UV-C light application and compared with that after a single application. Daily periodic preharvest UV-C light treatment showed a cumulative effect on grape stilbenoids. An 86-fold stilbenoid level increase (sum of E-resveratrol, E-piceatannol, ε-viniferin, E-piceid, isorhapontigenin, ω-viniferin, and Z-piceid) in grapes was achieved. The effects of UV-C light on stilbenoid in grape cane was also addressed for the first time. Stilbenoid oligomers such as hopeaphenol, ampelopsin A, and r-viniferin were quantified in cane samples. Quality grape parameters indicated an acceleration of ripening in UV-C samples.

Grapevine-shoot stilbene extract as a preservative in red wine.

This paper reports the use of a grapevine-shoot stilbene extract (Vineatrol®) as a preservative in red wine. Its effectiveness to preserve red wine quality under two different winemaking systems (traditional and Ganimede) was studied at bottling and after twelve months of storage in bottle. Enological parameters, color related parameters, volatile composition, sensory analysis and olfactometric profile were evaluated. At bottling wines treated with Vineatrol showed higher color related parameters and higher score in sensory analysis than those treated with SO2. The use of SO2 increased ester and alcohol volatile compounds in relation to the use of Vineatrol. Wine olfactometric profile was modified by Vineatrol addition. Two new odorant zones with high modified frequency appeared in wines treated with Vineatrol. After 12months of storage in bottle, wines treated with Vineatrol showed parameters related to oxidation. The weak point of the process seemed to be the evolution during the storage in bottle.

Preharvest methyl jasmonate and postharvest UVC treatments: increasing stilbenes in wine.

UNLABELLED: Stilbene-enriched wine is considered to be an interesting new food product with added value due to its potential health-promoting properties. Stilbene concentration in grape is highly variable and rather scarce. However, it can be increased by stress treatments. For this reason, numerous pre- and postharvest grape treatments, and some combinations of them, have been tested to maximize stilbene content in grapes. In the present manuscript, Syrah grapes were treated with (i) methyl jasmonate (MEJA), (ii) ultraviolet light (UVC), and (iii) methyl jasmonate and ultraviolet light (MEJA-UVC) and compared with untreated grapes. Afterward, winemaking was developed. Wine achieved by combination of both treatments (MEJA-UVC) contained significantly higher stilbene concentration (trans-resveratrol and piceatannol) than its respective control (2.5-fold). Wine quality was improved in color-related parameters (color intensity, L*, a*, b*, ΔE*, anthocyanins, and tannin). Moreover, MEJA-UVC wines obtained the highest score in sensorial analysis. To the best of our knowledge, this is the first time that pre- and postharvest treatments are combined to increase stilbenes in wine. PRACTICAL APPLICATION: The effect of treatment combination (methyl jasmonate and UVC light) on grape and wine was evaluated. Our results highlight the positive effect of the treatments in stilbene content, color parameters, and sensorial analysis. Moreover, added-value by-products were achieved.